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International Journal of Current Research and Review (IJCRR)
Vol 04 / Issue 8 / April 2012
Frequency: Fortnightly
Language: English
1
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Vol. 04 issue 08 April 2012
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ISSN 0975-5241
IC Value of Journal: 4.18
“Let the science be your passion”
Vol 4 / Issue 8 / April 2012
2
Editorial Board
Dr. Prof. Dato‘
Proom Promwichit
Dr. Nahla Salah Eldin
Barakat
Dr. Ann Magoufis
Dr. Pongsak
Rattanachaikunsopon
Dr. Chellappan
Dinesh
Dr. R. O. Ganjiwale
Dr. Shailesh Wader
Dr. Alabi Olufemi
Mobolaji
Dr. Joshua Danso
Owusu-Sekyere
Dr. Okorie
Ndidiamaka Hannah
Dr. Parichat
Phumkhachorn
Dr. Manoj Charde
Dr. Shah Murad
Mastoi
Deputy Vice Chancellor, Research & Innovation
Division, Masterskill University College of
Health Sciences, Cheras, Malaysia
Faculty, University of Alexandria, Alexandria,
Egypt
Director, Ariston College, Shannon, Ireland
Faculty, Ubon Ratchathani University, Warin
Chamrap, Ubon Ratchathani, Thailand
Dean, School of Pharmacy, Masterskill
University College of Health Sciences, Cheras,
Malaysia
HOD, Department of Pharmacognosy, I.P.E.R.
Wardha, Maharashtra
HOD, Department of Pharmaceutical Chemistry,
IPER, Wardha, MH, India
Faculty, Bowen University, Iwo, Osun-State,
Nigeria
Faculty, University of Cape Coast, Cape Coast,
Ghana
Faculty, University of Nigeria Nsukka, Enugu
State
Faculty, Ubon Ratchathani University, Warin
Chamrap, Ubon Ratchathani, Thailand
Dean, NRI Group of Post Graduate Studies,
Bhopal
HOD, Pharmacology and Therapeutics, Lahore
Medical and Dental College, Lahore, Pakistan
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About International Journal of Current Research and Review (ijcrr)
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Vol. 04 issue 08 April 2012
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Index
S.
N.
1
2
3
4
5
6
7
8
9
Title
4
Page
No.
A Study on Co Relation between
Straight Leg Angle and Functional
Disability in Low Back Pain
Abhishek Sharma,
Urvi Bhavsar
6
Use of Antimicrobial Prophylaxis in
Surgical and Medical Intensive Care
Units – A Comparative Study
Scenario
of
Biomedical
Waste
Management in the Major Hospitals of
Srinagar City
Patients‘ Perception on Hospital
Services with Special Reference to
Behavior of Doctors in Villupuram
District
Dna Barcoding, Phylogenetic Diversity
Studies of Etroplus Suratensis Fish from
Pooranankuppam
Brackish
Water,
Puducherry
Apurva Agrawal,
Barna Ganguly
10
Rumisa Nazir, G. A.
Bhat
16
D. Karthikeyan, M.
Thurunarayanasamy
23
Is Skeletal Muscle ‗A Target Organ‘ in
Long Term Uncontrolled Diabetes
Mellitus?
A
Comparative
and
Correlative study of Type I and Type II
Diabetes
In-Silico Studies on P43 Protein from
Plasmodium Falciparum
Comparative Microbiologic Analysis of
Subgingival Plaque Samples in Type II
Diabetic and Non – Diabetic Patients with
Chronic Periodontitis by Polymerase Chain
Reaction
Experimental Behaviour of a Pyramid
Type Solar Still Coupled and Decoupled
to an Electrical Temperature Controller
Vol 4 / Issue 8 / April 2012
10
Authors
Geomagnetic Storms Associated with
IV-Radio Bursts and their Relation with
Solar and Interplanetary Parameters
Sachithanandam V.,
Mohan P.M.,
Muruganandam N.,
Chaaithanya I.K.,
Arun Kumar P, Siva
Sankar R
Prathamesh Haridas
Kamble, Sunil
Bhamre
33
43
Tarun Kumar Bhatt
49
Mythireyi D, M G
Krishnababa,
Kalaivani
S.Kalaivani,
S.Rugmini
Radhakrishnan,
B.Selvakumar,
M.Indhumathy
P.L. Verma
55
63
77
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Vol. 04 issue 08 April 2012
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12
13
14
15
16
17
Phytochemicals Analysis of Colecus
Arometicus Benth
Toxicological Evaluation of Aqueous
Leaf Extract of Senna Alata in Pregnant
Wistar Rats
A Comparative Study to assess the
Changes in the Conduction of Median
Nerve at Wrist Joint in apparently
asymptomatic computer users with that
in general population
Design of a Compact CPW Fed
Hexagon Shaped Slot Antenna for WiMax Application
Bioequivalence and Highly Variable
Drugs: An Overview
An Economic Analysis of
Diversification in Tamil nadu
Crop
Page
No.
Iffat Khan, Kirti Jain
85
Yakubu M. T.,
Adeshina A. O.,
Ibrahim, O. O. K.
Dhaval Desai, Chintan
Shah, Harshit Soni2,
Hasmukh Patel1,
Komal Soni
H. M. Ramesh, K.
Balaji, D.Ujwala,
B.Harish, Ch. Vijay
Sekhar Babu, K.Naga
Mallik
Vikram Lohar, Harsh
Patel, Arvind Singh
Rathore, Sandeep
Singhal, Ashish
Kumar Sharma, Parul
Sharma
89
110
119
124
V. Kalaiselvi
147
Chintan Pathak, V. K.
Srivastava
155
18
Wireless Technology in Service of
SocietyA Case Study Of Snakebite
P.P.Patil, Abhijit
A.Patil, B T Jadhav
168
19
Production and Optimization of Single
Cell Oil by Oleaginous Bacteria Isolated
from Oil Contaminated Environments
Study of the Reasons of the
Radiographic Images Repetition in
Sistan and Baluchestan‘s Treatment
Centers
T. Murugan, D.
Saravanan,
R.Balagurunathan
Mohammad Javad
Keikhai Farzaneh,
Reza Afzalipour,
Mojtaba Vardian,
Mahdi Shirin Shandiz,
Mohammad zarei
20
5
Authors
Oil-Water Separation using Fly Ash
Zeolite Treatment
“Let the science be your passion”
Vol 4 / Issue 8 / April 2012
Title
175
185
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
A STUDY ON CO RELATION BETWEEN STRAIGHT LEG
ANGLE AND FUNCTIONAL DISABILITY IN LOW BACK
PAIN
Abhishek Sharma1, Urvi Bhavsar2
ijcrr
Vol 04 issue 08
Category: Research
Received on:28/01/12
Revised on:17/02/12
Accepted on:07/03/12
1
2
SPB Physiotherapy College, Surat,Gujarat
Masoom Hospital, Surat, Gujarat
E-mail of Corresponding Author: drabhi2700@gmail.com
ABSTRACT
Back ground and Purpose: Low back Pain is a major cause of functional disability in India. It is because
of trauma, degeneration or any pathology related to back. Low back pain is the most challenging job of
physiotherapist to deal with it. The aim of the study was to study SLR angle, the angle at which the pain
increases in severity or radiating starts, and functional disability. The second was to know severity of
disability in routine functions of low back pain patients. Research Design: Co-relation method using
Karl Pearson. Material and Methods: Personal interview of 30 patients with Low Back Pain was taken
by using to know about the level of functional disabilities and Straight Leg Raise (SLR) angle was
measured. Co-relation between both Roland and Morris questionnaire and Straight Leg Raise (SLR)
Results: The co-relation between SLR angle and functional disability as measured by RM questionnaire
is partial negative. The co-relation co-efficient is -0.75. Conclusion: SLR angle is indicative of the
functional disabilities. The patient with more disability has less SLR angle
____________________________________________________________________________________
Key Words: Roland and Morris questionnaire,
Straight Leg Raise (SLR) angle, Low back Pain
INTRODUCTION
Low back pain or lumbago is a common
musculoskeletal disorder affecting 80% of
people at some point in their lives. It is an
extremely common human phenomenon which
occurs because of trauma, degeneration or any
pathology related to back. It can be either acute,
sub acute or chronic in duration19. Low back
pain (LBP) is defined as chronic after 3 months,
unless pathoanatomic instability persists. A
slower rate of tissue repair in the relatively
avascular intervertebral disk may impair the
resolution of some persistent painful cases of
chronic LBP (cLBP) 18. The lumbar region (or
6
lower back region) is made up of five vertebrae
(L1-L5)5,8,17. In between these vertebrae lie fibro
cartilage discs (intervertebral discs), which act
as cushions, nerves runs from the spinal cord
through foramina within the vertebrae, providing
muscles with sensations and motor associated
messages. Stability of the spine is provided
through ligaments and muscles of the back,
lower back and abdomen. Small joints those
prevent as well as direct motion of the spine are
called facet joints (zygapophysial joints).Low
back pain is more persistent among people who
previously required time off from work because
of low back pain, those who expect passive
treatments to help, those who believe that back
pain is harmful or disabling or fear that any
movement whatever will increase their pain, and
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
people who have depression or anxiety3,4.This
dysfunction leads to disability to perform any
household activites, grooming and almost all
daily activities. A disability is any long-term
limitation in activity resulting from a condition
or health problem. Disability mainly occurs in
performing daily activities as dressing,
grooming, lifting, etc.
Purpose of the Study
The first aim of the study was to study SLR
angle, the angle at which the pain increases in
severity or radiating starts, and functional
disability. The second was to know severity of
disability in routine functions of low back pain
patients.
METHODOLOGY
Research Design: Simple Random Sampling
Method .Co-relation method using Karl Pearson
Source of Data:
Patients with pathological
back pain
Inclusion Criteria: Age group: 30 to 65 years
of age.Sex: Male and female.
Patients with pathological back pain like
herniated
disc,
spinal
Stenosis,
spondylolisthesis, Sciatica, etc.
Exclusion Criteria: Patients with mechanical
back pain, osteoporosis, any spinal deformity,
Ankylosis spondilitis.
Sampling Technique and Sample Size
All subjects who fulfilled the inclusion criteria
were selected for the study which counts to a
total of 30 patients
Data Collection Method
Personal interview method by using Roland and
Morris questionnaire and measurement of
Straight Leg Raise (SLR) angle.
Outcome measures:
RM (Roland and Morris questionnaire) scale
was used to measure score of RM index.SLR
7
angle was used to measure at which pain
reproduces or radiating pain starts.
Collection of Data
Roland and Morris developed a questionnaire
for evaluating patients with low back pain1,16.
This can be used to determine the level of
patient disability and can help measure outcome
following
therapeutic
intervention.
The
Questionnaire is usually paired with a visual
analogue scale (VAS) for pain rating ranging
from no pain at all to unbearable pain. There are
total 24 questions, each has 1 point. If the
answer is yes the score is 1 and for no score is
0.Total score = SUM (Points for all 24
statements).Interpretation-Minimum score: 0
Maximum Score: 24. The higher the score the
more severe the disability associated with low
back pain. A score of 0 indicates no disability
and 24 severe disabilities. A score ≥ 14 indicates
a patient in poor outcome group. Explain the
patient the ROLAND AND MORRIS
QUESTIONNAIRE. As they read a sentence
that describes them today, put tick against it. If
sentence does not describe them, then leave the
space blank and go on to the next one.
STRAIGHT LEG RAISING (SLR) 2,4,5,6,19:
Unilateral SLR test test is positive if pain
extends from the back down into the leg in the
sciatic nerve distribution. With the patient in
supine position, the hip is medially rotated and
adducted, the knee is extended, and the examiner
flexes the hip until the patient complains of the
pain or tightness in back region or back of the
leg. With the patient in supine position, the
therapist passively flexes the hip until the
symptom comes, this angle is measured by
Goniometer.The fulcrum of goniometer is
placed at greater trochanter; the moveable arm
of the goniometer is placed along the shaft of the
femur. The angle between fixed arm and
movable arm is measured. The score of every
patient and SLR angle should be noted and
analyzed.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
DATA ANALYSIS
The collected data were subjected to Karl
Pearson Correlation Co-efficient method.
RESULTS
The co-relation co-efficient is -0.75.It shows
moderately or partial negative co-relation
between functional disability and SLR angle
DISCUSSION
There is partial negative co-relation between
SLR and functional disability. That means, both
variables are inversely related to each other. As
a functional disability is more, the SLR angle is
less. The pain produces at a lower angle because
of more disability .The study also relates low
back pain and functional disability. We can
make the society aware about the disabilities
9,10,11,12,14
due to the back pain and explain them
to take care about the daily activities. Low back
pain is the most common problem in this era for
physiotherapist to deal with.
CONCLUSION
The study concluded that there is a partial
negative
correlation between functional
disability and SLR angle that is patient with
more disability has less SLR angle.SLR
ACKNOWLEDGEMENTS
We are sincerely grateful to the god who
showered blessings for our research. We are
thankful to our principal for his help in every
walk of our research. We would like to mention
special thanks to my entire faculty who were
always a standing rock for our great work. We
are privilege to thank our patients without whom
this research was not been possible. We
acknowledge the immense help received from
the scholar whose articles are cited and included
in references of this manuscript. We are grateful
to authors/editors/publishers of all those articles,
8
journals and books from where the literature for
this research article has been reviewed
ETHICAL COMMITTEE CLEARANCE:
There was no ethical committee formed in the
institution during the time in which research was
performed.
CONFLICT OF INTEREST: None.
1.
2.
3.
4.
5.
6.
7.
8.
REFERENCES
Brouwer S., Reliability and stability of
Roland
and
Morris
Disability
Questionnaire was checked. Disability
2004 Feb; 26(3):162-5.
Capra F, Vanti C, Donati R, Tombetti S,
O'Reilly C, Pillastrini P.Validity of straight
leg raise with sciatic pain with or without
lumbar pain using Magnetic resonance
imaging as standard Reference. Journal of
Manipulative
and
Physiological
Therapeutics. 2011; 34(4):231-8.
Krause N, Ragland DR.Occupational
disability due to low back pain: a new
interdisciplinary Classification based on a
phase model of disability. Spine. 1994
May; 19(9):1011-20.
Lutz GK, Butzlaff M, Schultz-Venrath U.
Looking back on back pain: trial and error
of diagnoses in the 20th century. Spine
.1976 Aug.; 28(16):1899-905.
Bernard E.Finneson.Low Back Pain.
McMinn Publishers.1998; 2nd Edition.
Atul Patel, ABNA A. OGLE. Diagnosis
and Management of Low Back Pain.
Family
Physician. 2000 Mar 15;
61(6):1779-1786
Stratford, P.W., Binkley, J.M., & Riddle,
D.L.Development and initial validation of
the back pain functional scale. Health
Services Research, 2000. 25 (16), 20952102
B.D.Chaurasia.Clinical
Anatomy.CBS
Publishers; 1996,3rd edition.
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9.
10.
11.
12.
13.
Debbie Ehrmann Feldman. Risk Factors for
the Development of Low Back Pain in
Adolescence. Oxford Journals, American
Journal of Epidemiology: 2000.154(1); 3036
Robbert
Braton.Assessment
and
Management of Acute Low Back Pain.
America Physician. 1999; 60(8):22992306.
Scientific approach of the assessment and
management of activity-related spinal
disorders. A Monograph for clinicians.
Report of the Quebec Task Force on Spinal
Disorders. Spine. 1987; 12(7):S1–59.
BK
Mahajan.Methods
in
Biostatistics.Jaypee; 6th Edition.
Maurits van Tulder.Exercise Therapy for
Low Back Pain; A Systematic Review
Within the Framework of the Cochrane
Collaboration Back Review Group .SPINE.
2000.; 25(21): 2784 –2796.
9
14. A Kopec The Quebec Back Pain Disability
Scale: conceptualization and development.
Journal of Clinical Epidemiology.1996. ;
49(2):151-161.
15. John Fran. Preventing disability from
work-related low-back pain. Canadian
medical journal.1998 ; 158:1625-3.
16. Stratford PW et al. A comparison study of
the back pain functional scale and Roland
Morris Questionnaire. North American
Orthopedic
Rehabilitation
Research
Network. Journal of Rheumatology. 2000
Aug; 27(8):1928-36.
17. Web
supportswww.pubmed.com.www.google.com.
18. A. F. Mannion, M. Müntener.et
al.Comparison of three active therapies for
chronic low back pain: results of a
randomized clinical trial with one‐ year
follow‐ up.Journal
of
Rheumatology
(2001) 40 (7): 772-778.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
USE OF ANTIMICROBIAL PROPHYLAXIS IN SURGICAL
AND
MEDICAL
INTENSIVE
CARE
UNITS
–
A
COMPARATIVE STUDY
Apurva Agrawal1, Barna Ganguly2
ijcrr
Vol 04 issue 08
Category: Research
Received on:03/03/12
Revised on:16/03/12
Accepted on:22/03/12
1
Department of Pharmacology, Geetanjali Medical College, Udaipur, Rajasthan
Department of Pharmacology, Pramukhswami Medical College, Karamsad, Gujarat
2
E-mail of Corresponding Author: apuragr@yahoo.co.in
ABSTRACT
Aims: To study the extent and pattern of use of antibiotics for prophylaxis in medical and surgical
intensive care units. Subjects and Methods: 100 patients each from SICU and MICU were included in
the study. Case record files were analyzed daily until discharge from ICU or a maximum of 21 days.
Details of all antibiotics prescribed for prophylaxis were recorded in a proforma, which were then
analyzed using relevant statistical tests. Results: 65% patients in MICU and 99% in SICU received
antibiotics (p value <0.0001). Among patients who received antibiotics, 37% in MICU and 73% in SICU
received them for prophylaxis (p value <0.0001). Average duration of prophylaxis was 2.58 days in
MICU and 3.14 days in SICU. 19 (79.14%) patients in MICU and 48 (66.67%) patients in SICU received
prophylaxis for more than 24 hours (p value = 0.3690). 15 (62.5%) patients in MICU and 31 (43%)
patients in SICU received combination of antibiotics for prophylaxis (p value = 0.156). Third generation
cephalosporins were the most commonly prescribed antibiotics for prophylaxis in both ICUs.
Conclusion: Widespread use of antimicrobial prophylaxis in ICUs with broad spectrum antibiotics and
antibiotic combinations, with duration longer than recommended has emerged as area of concern in
present study. Such surveillance studies help in recognition of areas requiring special attention, which can
guide the formulation of antibiotic prescription policies at individual ICU level.
____________________________________________________________________________________
Keywords: Antibiotic, antimicrobial resistance,
ICU.
INTRODUCTION
Infections are a frequent problem in Intensive
Care Units (ICUs) and thus antibiotics are
frequently used. Although antibiotics represent
one of the most frequently prescribed classes of
drugs among all hospitalized patients, total
antibiotic consumption is much higher in the
ICU than in general hospital wards. [1] Besides
treatment of infections, antibiotics in ICU are
administered as prophylaxis to prevent or limit
10
major infections in critically ill patients. [2]
Antibiotic prophylaxis is highly effective in
some clinical settings, but in others, it accounts
for misuses of antimicrobials, and may even be
deleterious. [3] A number of studies have justified
antibiotic prophylaxis in dirty or contaminated
surgical procedures, where the incidence of
wound infections is high, but such use must not
be extended beyond 24 hours. [3] These include
less than 10% of all surgical procedures. In clean
surgical procedures, which account for
approximately 75% of the total, antibiotics
should not be routinely used as the expected
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
incidence of wound infection is less than 5%.
Except a very few conditions, non surgical
antibiotic prophylaxis is not routinely indicated.
Although awareness of the consequences of
antibiotic misuse is increasing, overprescribing
remains widespread. Overuse of antibiotics and
poor compliance with infection control measures
have been identified as the two major reasons
for increasing antimicrobial resistance. [4]
Studies on antibiotic prescription practices in
ICUs have been done in some countries, [1,5] but
information regarding studies done in Indian
ICU setting is extremely limited. Antibiotic
recommendations based on studies performed at
a few selected centers may not be applicable and
may not be generalized to all ICU settings.
Singh N et al has suggested that research in
individual ICU is essential in guiding antibiotic
prescription practices. [6] As antibiotic policy for
ICU in our Hospital was in developmental phase
and we needed to know the potential areas of
concern, a cross sectional study was done, to
study the extent and pattern of use of antibiotics
for prophylaxis in medical and surgical intensive
care units.
MATERIAL AND METHODS
The Hospital, in which this study was
conducted, is a 550 bedded tertiary care hospital.
There are two adult ICUs, medical and surgical
ICU, each ICU is a 12 bedded unit. In surgical
ICU (SICU) majority of patients admitted are
because of road traffic accidents and surgical
patients admitted after various surgical
procedures. In medical ICU majority of patients
admitted are because of respiratory, cardiac or
multiorgan failure.
Inclusion and exclusion criteria:
Patients admitted in SICU and MICU, of age
above 18 years, irrespective of sex were
included in the study from October 2007 to
October 2009. Patients whose relatives were not
11
willing to give consent and patients with age less
than 18 years were excluded from the study.
Collection of data:
Permission was taken from Institutional Human
Research Ethics Committee before starting the
study. A total of 200 patients were included
randomly, 100 from each ICU. As patients
admitted in ICU are critically ill, written
informed consent was taken from the relatives of
the patients. Case record files of the included
patients were analyzed daily until their discharge
from ICU or a maximum of 21 days. Treatment
was considered prophylactic if there was no
evidence of infection and the drug was used to
prevent infection. Details of all antibiotics
prescribed for prophylaxis were recorded in a
predecided proforma, including group of drug,
route of administration, dose and duration of
treatment.
Statistical analysis:
Data was analysed on Microsoft excel 2007.
Mean and frequencies were calculated in the two
groups. Chi square test was used to compare
proportions and p value of < 0.05 was
considered as significant.
RESULTS
In MICU 65 (65%) patients were prescribed
antibiotics either single or in combination while
in SICU 99 (99%) patients were prescribed
antibiotics (p value <0.0001). In MICU
antibiotics consisted 15% of total drugs
(excluding intravenous fluids) prescribed while
in SICU 23.54% of total drugs prescribed (p
value <0.0001). Out of 65 patients who received
antibiotics in MICU, 24 (37%) received them for
prophylaxis, while in SICU 72 (73%) out of 99
patients received antimicrobial prophylaxis (p
value <0.0001). In MICU 4.6% received
surgical prophylaxis and 33.8% received non
surgical prophylaxis. In SICU 37% received
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
surgical prophylaxis and 36% non surgical
prophylaxis.
Average duration of antimicrobial prophylaxis in
MICU was 2.58 days and in SICU 5.24 days.
Antimicrobial prophylaxis for more than 24
hours was given in 19 (79.14%) patients in
MICU and in 48 (66.67%) patients in SICU (p
value = 0.3690). In 13 (54.2%) patients in
MICU and in 32 (44.4%) patients in SICU
antimicrobial prophylaxis was used for more
than 48 hours (p value = 0.5549).
Third generation cephalosporins were the most
commonly prescribed antibiotics for prophylaxis
in both ICUs (62.5% and 83.33% in MICU and
SICU respectively) followed by metronidazole
(45.83%) in MICU and metronidazole (19.44%)
and amikacin (19.44%) in SICU. Frequency of
different groups of antibiotics used for
prophylaxis in MICU and SICU is given in
Table I & II respectively. Comparison of most
commonly used antibiotic groups in both ICUs
is given in Table III.
Out of those patients who received antimicrobial
prophylaxis, 15 (62.5%) patients in MICU while
31 (43%) patients in SICU received combination
of antibiotics (p value = 0.156). Most common
combination used in MICU was 3rd generation
cephalosporin with metronidazole, while in
SICU 3rd generation cephalosporin with
aminoglycoside was the most common
combination used for prophylaxis.
Among patients who received antibiotics for
prophylaxis, 6 (25%) patients in MICU and 22
(30.55%) patients in SICU, later received
empirical therapy for suspected infection despite
of antimicrobial prophylaxis (p value = 0.7954).
DISCUSSION
As patients in ICUs are critically ill and more
susceptible to nosocomial infections, more
frequent use of antibiotics in these units is
expected. In a study done by Roder BL et al,
total antibiotic consumption was approximately
12
ten times greater in ICU wards than in other
wards. [1] In our study also, antibiotics were
frequently prescribed, 65% patients in MICU
and 99% in SICU received antibiotics. Though
use of antibiotics in MICU was in accordance
with that reported by Roder BL et al and
Bergmans DCJJ et al in their studies, [1,7]
antibiotic use in SICU was far more prevalent,
much higher than found in other studies. [1,7,8]
Similarly prophylactic use of antibacterials was
significantly less in MICU as compared to
SICU; it was also less than reported by studies
done in other countries, [1,5] where more than half
of the patients received antimicrobial
prophylaxis. The picture in SICU was different,
where out of total patients who received
antibiotics,
73%
received
antimicrobial
prophylaxis, a figure much higher than found in
above mentioned studies. The incidence of non
surgical prophylaxis was almost similar in the
two ICUs, as more surgical patients are admitted
in SICU; surgical prophylaxis is mostly
responsible for this difference.
Many guidelines are available for surgical
prophylaxis which recommend 1st generation
cephalosporin as first choice and for not more
than 24 hours. [3,9] Concerning non-surgical
prophylaxis, excluding a few specific conditions
like neutropenia, there is evidence for only two
approaches that are oral decontamination and
selective digestive decontamination (SDD). [10-12]
Observations in our study were far away from
these
recommendations.
Inappropriate
antibiotics were prescribed for lengthy periods
(mean duration 2.58 days in MICU and 3.14
days in SICU), and in too many patients. Instead
of 1st generation cephalosporins which are
recommended for surgical prophylaxis, 3rd
generation cephalosporins were the most
commonly used antibiotics in both ICUs.
Antibiotic combinations were frequently used in
both ICUs for prophylactic purpose. Providing
broad spectrum coverage to patients might be a
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
reason for high prevalence of use of antibiotic
combinations. Oral decontamination and SDD
were never used in this study for non surgical
prophylaxis. Use of antibiotic prophylaxis in
non-surgical patients except a few specified
conditions is not recommended by any
guidelines. This practice increases the chances
of development of antibiotic resistance and
induces a false sense of confidence in clinicians
who consequently pay less attention to the
possibility of occult infection. [5] This possibility
correlates with the finding in our study that out
of total patients who received antimicrobial
prophylaxis, 6 (25%) patients in MICU and 22
(30.55%) patients in SICU later received
empirical antibiotics for suspected infection
despite of antimicrobial prophylaxis. Adherence
to internationally accepted guidelines has been
found low in other studies also. [5,13]
There were some limitations in present study.
Relatively small number of patients was studied
in both intensive care units. As clinicians were
not included in the present study frame, we can
not conclude on the reasons responsible for
nonadherence of antibiotic prescription practices
to internationally accepted guidelines.
CONCLUSION
In the current scenario when antimicrobial
resistance is growing in intensive care units and
nosocomial infections are becoming more and
more difficult to treat, appropriate and cautious
use of antibiotics particularly in intensive care
units becomes a necessity so that we can use
these wonder drugs in future also. Present study
has provided a baseline data of the prophylactic
use of antibiotics in intensive care units of a
tertiary care teaching hospital. Liberal use of
antibiotics for surgical and non surgical
prophylaxis, with broad spectrum antibiotics and
antibiotic combinations, and for long durations,
has emerged as areas of concern in present
study. As characteristics of patient population,
13
their risks and susceptibilities to various
infections, as well as predominant pathogens and
their antimicrobial resistance varies between
different ICUs, such surveillance studies and
research help in recognition of areas of special
concern which can guide the formulation of
antibiotic prescription policies at individual ICU
level.
ACKNOWLEDGEMENT
We acknowledge the immense help received
from the scholars whose articles are cited and
included in references of this manuscript. We
are also grateful to authors / editors / publishers
of all those articles, journals and books from
where the literature for this article has been
reviewed and discussed.
REFERENCES
1. Roder BL, Nielsen SL, Magnussen P,
Engquist A, Frimodt-Moller N. Antibiotic
usage in an intensive care unit in a danish
university hospital. J Antimicrob Chemother
1993;32:633-42.
2. Mangram AJ, Horan TC, Pearson ML,
Silver LC, Jarvis WR. Guideline for
prevention of surgical site infection, 1999.
Hospital Infection Control Practices
Advisory Committee. Infect Control Hosp
Epidemiol 1999;20(4):250-78.
3. Chambers HF. General principles of
antimicrobial therapy. In: Brunton LL,
editor. Goodman & gilman‘s the
pharmacological basis of therapeutics. 11th
ed. New York: McGraw-Hill; 2006. p. 1095110.
4. Goldmann DA, Weinstein RA, Wenzel RP
et al. Strategies to prevent and control the
emergence and spread of antimicrobialresistant microorganisms in hospitals. a
challenge to hospital leadership. JAMA
1996 Jan 17;275(3):234–40.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
5. Malacarne P, Carlotta R, Bertolini G.
Antibiotic usage in intensive care units: a
pharmaco-epidemiological
multicentre
study.
J
Antimicrob
Chemother
2004;54(1):221-4.
6. Singh N, Yu VL. Rational empiric antibiotic
prescription in the ICU – clinical research is
mandatory. Chest 2000;117(5):1496-9.
7. Bergmans DCJJ, Bontena MJM, Gaillardc
CA et al. Indications for antibiotic use in
ICU patients: a one-year prospective
surveillance. J Antimicrob Chemother
1997;39:527-35.
8. Hartmann B, Junger A, Brammen D et al.
Review of antibiotic drug use in a surgical
ICU: management with a patient data
management system for additional outcome
analysis in patients staying more than 24
hours. Clin Ther 2004 June;26(6):915-24.
9. Lampiris HW, Maddix DS. Clinical use of
antimicrobial agents. In: Katzung BG,
editor. Basic and clinical pharmacology.
11th ed. Boston Burr Ridge (IL): The
McGraw-Hill Companies; 2009. p. 827-41.
10. Marino PL. The ICU book. 3rd ed.
Philadelphia (PA), USA: Lippincott
Williams & Wilkins; 2007. p. 63-80.
11. Bergmans DCJJ, Bonten MJM, Gaillard CA
et al. Prevention of Ventilator-associated
Pneumonia by Oral Decontamination. A
Prospective, Randomized, Double-blind,
Placebo-controlled Study. Am J Respir Crit
Care Med 2001 Aug;164(3):382-8.
12. Ulrich C, Harinck-de Weerd JE, Bakker NC,
Jacz K, Doornbos L, de Ridder VA.
Selective decontamination of the digestive
tract with norfloxacin in the prevention of
ICU-acquired infections: a prospective
randomized study. Intensive Care Med
1989;15(7):424-31.
13. van Kasteren MEE, Kullberg BJ, de Boer
AS, Mintjes-de Groot J, Gyssens IC.
Adherence to local hospital guidelines for
surgical antimicrobial prophylaxis: a
multicentre audit in Dutch hospitals. J
Antimicrob Chemother 2003;51:1389-96.
Table I: Group wise distribution of antibiotics used for prophylaxis in MICU (n = 24)
S. No.
1.
2.
3.
4.
5.
6.
Group of antibiotic
(%)
Penicillin
(41.66%)
Cephalosporin
(62.50%)
Fluoroquinolone
(4.17%)
Aminoglycoside
(4.17%)
Tetracycline
(12.50%)
Other
(45.83%)
Name / Generation of antibiotic
Amoxicillin
Amoxicillin-Clavulanate
Ampicillin
Cloxacillin
III Generation Cephalosporin
No. of patients
(%)
4 (16.67)
2 (8.33)
2 (8.33)
2 (8.33)
15 (62.50)
Levofloxacin
1 (4.17)
Gentamicin
1 (4.17)
Doxycycline
3 (12.50)
Metronidazole
11 (45.83)
* Some patients received more than one drug, and therefore the total percentage exceeds
100%.
14
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table II: Group wise distribution of antibiotics used for prophylaxis in SICU (n = 72)
S. No.
1.
2.
3.
4.
5.
6.
Group of antibiotic
(%)
Penicillin
(13.89%)
Cephalosporin
(83.33%)
Fluoroquinolone
(20.83%)
Aminoglycoside
(27.77%)
Antifungal
(1.39%)
Other
(22.22%)
Name / Generation of antibiotic
Amoxicillin – Clavulanate
Ampicillin – Sulbactum
Cloxacillin
Piperacillin – Tazobactum
II Generation Cephalosporin
III Generation Cephalosporin
Levofloxacin
Ciprofloxacin
Ofloxacin
Amikacin
Neosporin
Fluconazole
Metronidazole
Teicoplanin
No. of patients
(%)
1 (1.39)
1 (1.39)
6 (8.33)
2 (2.78)
6 (8.33)
54 (75.00)
10 (13.89)
4 (5.55)
1 (1.39)
14 (19.44)
6 (8.33)
1 (1.39)
14 (19.44)
2 (2.78)
* Some patients received more than one drug, and therefore the total percentage exceeds 100%
Table III: Comparison of frequency of various antibiotics used for prophylaxis in MICU & SICU
S. No.
1.
2.
3.
4.
5.
Name of Antibiotic
Penicillins
Cephalosporins
Fluoroquinolones
Aminoglycosides
Metronidazole
MICU (n=24)
10
15
1
1
11
SICU(n=72)
10
60
15
20
14
P value
0.0090
0.0639
0.1138
0.0325
0.0225
* P value calculated by chi square test
15
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
SCENARIO OF BIOMEDICAL WASTE MANAGEMENT IN
THE MAJOR HOSPITALS OF SRINAGAR CITY
Rumisa Nazir, G. A. Bhat
ijcrr
Vol 04 issue 08
Category: Research
Received on:18/02/12
Revised on:03/03/12
Accepted on:19/03/12
P. G. Department of Environmental Science / Centre of Research for Development,
University of Kashmir, Srinagar, J & K
E-mail of Corresponding Author: Rumisanazir@yahoo.com
ABSTRACT
In order to assess the biomedical waste management; the practices currently operative and compliance
with Regulatory Notification for Biomedical Waste (Management and Handling) Rules, 1998, under the
Environment (Protection Act 1986), Ministry of Environment and Forestry, Govt of India; the level of
awareness regarding biomedical waste management and handling rules among the hospital staff; training
imparted to the waste handlers and other particulars regarding risk associated with the handling of
biomedical waste, the present study was carried out during May-July 2010 which involved the use of
questionnaire method, in-depth interview and personal observation to crosscheck the authenticity of
information gathered. During the study, the existing practices of biomedical waste management appeared
to be unsatisfactory; hospitals did not conform to the Biomedical Waste (Management and Handling)
Rules, 1998. Waste segregation was found not practiced by the hospitals surveyed and knowledge
regarding biomedical waste management was found highest among the doctors i.e. 94.3% and 96% at
SKIMS and SMHS hospital respectively indicating that people with higher qualification possessed more
awareness regarding the prescribed rules. No specific training and awareness programs on biomedical
waste management were organized by the hospital authorities.
____________________________________________________________________________________
Keywords: Biomedical waste, Segregation,
Knowledge, Training, Hospital, existing
practices.
INTRODUCTION
Hospitals are service oriented institutes that
provide medical facilities vital for our life and
health. In the healthcare process, waste is
generated which usually includes sharps, human
tissues or body parts and other infectious
materials (Baveja et al., 2000), also referred to
as Hospital Solid Waste‖ and ―Biomedical
Waste‖ (Manohar et al., 1998). It is a real
problem of living nature and human world as it
16
carries a higher potential for infection than any
other type of waste. Waste is an unavoidable
byproduct of human activities, pervading our
environment for centuries and will continue to
contaminate it. Therefore it is essential to have
safe and reliable methods for waste
management, focussing both on effective
training and supervision. Waste management
includes responsible planning of collecting,
transporting, processing, and disposing waste
material (Campbell, 1988; Stamenkovic, 2007).
Within waste management, the healthcare waste
management is a process that helps to ensure
proper hospital hygiene and safety of healthcare
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
workers and communities (Belkin et al.,1982;
Baram, 1989).The primary sources of
biomedical waste are hospitals, laboratories,
diagnostic centres, blood banks, veterinary
hospitals, nursing homes, clinics. Noninfectious waste forms nearly 85% of the waste
generated by a hospital and the remaining 10%
are hazardous (Pruss and Townend, 1998).
Inappropriate and inadequate handling of
biomedical waste may have a serious and
significant impact on the public health and the
environment. Sound management of biomedical
waste needs to be given priority and made an
integral feature of healthcare services. There is a
need to sensitize the top level waste managers
by making them aware of not only the various
types of waste, but also its generation,
collection, containment, handling, storage,
transportation, treatment and final disposal. The
proper segregation at source is an essential
element of the successful waste management
programme (pandit, 1999). If the infectious
component gets mixed with the general noninfectious waste, the entire mass becomes
potentially infectious (Nugget, 2010).
Waste management has become a critical issue
both at national and international level. In July
1998, the Government of India Environment
Protection Act 1986 (Rule 29 of 1986) issued a
Notification on Biomedical Waste (Management
and Handling), Rules 1998, indicating the rules
for the management and handling of biomedical
waste. It defined ―biomedical waste‖ as any
waste, which is generated during the diagnosis,
treatment or immunization of human beings or
animals or in research activities pertaining
thereto or in the production or testing of
biological and including categories mentioned in
Schedule I (1998).
Looking into the existing scenario of biomedical
waste management in the country it was planned
to undertake a study to assess the current
practices of biomedical waste management and
17
its compliance with Regulatory Notification for
Biomedical Waste (Management and Handling)
Rules, 1998, under the Environment (Protection
Act 1986), Ministry of Environment and
Forestry, Govt. of India; the level of awareness
among the hospital staff; training imparted to the
waste handlers and other particulars regarding
risk associated with waste handling at two major
hospitals of Srinagar, Kashmir known for their
advanced diagnostic and surgical specialties.
The study lasted for a period of 3 months.
MATERIAL AND METHODS
Data regarding the current biomedical waste
management practices, level of awareness
regarding biomedical waste management and
handling rules prescribed therein was collected
by the questionnaire method. The design of the
questionnaire was based on the survey
questionnaire of World Health Organization
(WHO) with editorial changes and was framed
according to the objective needs of the study. It
was served to hospital administrators, doctors,
nurses, sanitary staff and hospital engineers.
Onsite personal observation of the management
practices were carried out for confirmation and
as a supplement to information gathered by the
questionnaire.
Formal interview was conducted to find whether
the training is being imparted to the waste
handlers and other particulars regarding risk
associated with the waste handling. Authenticity
of information so obtained was crosschecked
through personal observation. The study was
conducted with the prior approval of the subjects
and institutions.
STUDY AREA
The historical Srinagar city is the summer
capital of Jammu And Kashmir State,
surrounded by hills on east and north eastern
side loc
E
longitude. Altitude of Srinagar varies from 1580
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
m in the low lying vicinity of River Jhelum and
1620m on the eastern hill slopes with an average
elevation of about 1586m above mean sea level
(Bates, 2005). The city lies on both side of river
Jhelum, which swirls through the heart of the
Srinagar city.
For the present study two hospitals were selected
which have different characteristics in terms of
their size, treatment technology and the type of
patients catered. The two study stations were:
Sheri Kashmir Institute of Medical Sciences
(SKIMS), Soura
It is a tertiary care hospital, catering to the
average socio-economic class of people and
provides a total of 600 beds.
Shri Maharaja Hari Singh Hospital (SMHS),
Karan Nagar
It is a teaching hospital associated to
Government Medical College, Srinagar and is
the biggest general hospital in terms of bed
capacity (750).
RESULTS
biomedical waste
Current
management
practices
The important inferences regarding the various
components of the waste management hierarchy
like segregation, packaging, storage, collection,
transportation and disposal were drawn and then
the framework of compliance was assessed.
Barring SKIMS, the waste was not segregated at
source as prescribed in the Biomedical Waste
Management Rules, 1998 (Table 1). Due to poor
segregation practices, the general waste gets
mixed up with the infectious waste. Hospitals
were using uncovered plastic bins for waste
collection provided with same kind of color
coded labelled polybags. Polybags were not
sealed properly and its integrity was found not to
be preserved. In SMHS hospital, waste sharps
were contained without being subjected to
disinfection in open trays or in any of the bins.
18
While in SKIMS, sharps were disinfected
properly and finally incinerated.
Waste storage area at SKIMS was of a size
appropriate to the quantities of waste produced
but did not have secured bins to eliminate the
possibility of access to the waste by rodents,
flies, or other natural scavengers. The waste was
placed in an open area before disposal at SMHS
hospital, so it was easily accessible to
unauthorized personnel and animals. The
transportation of waste to the storage site was
done manually in SMHS hospital, while in
SKIMS trolleys and pipeline system (Chute) was
employed.
Biomedical waste was autoclaved and
incinerated (Type Brick Kiln; capacity 125
kg/hr) onsite, at SKIMS. Ash so obtained was
buried in onsite ash pits, neither lined from
below nor sealed above. Liquid waste was
treated in the treatment plant and flushed into
the sewers (Fig.1). SMHS hospital treats its
biomedical waste at Common Biomedical Waste
Treatment Facility (CBWTF), Lassipora
Pulwama, Kashmir, while the liquid waste was
flushed directly into the River Jhelum.
The level of awareness among the hospital
staff
Knowledge regarding biomedical waste
management and rules prescribed therein at
SKIMS and SMHS hospital was found highest
among the doctors in the tune of 94.3% and 96%
respectively. Sanitary staff scored the least
which indicates that the authority neither
informed them in the form of instructions nor
did they supervised their biomedical waste
management practices (Fig. 2).
Training and other particulars regarding risk
in the waste handling
Although 10% waste handlers were aware of
risks involved in biomedical waste handling,
7.27% had received special training on this
aspect. While 18.18% of waste handlers suffered
from skin infections but none reported it to the
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
higher authorities. 3.63% of waste handlers
stated about being immunized (Table 2).
Out of 100 waste handlers which were
interviewed at SMHS hospital, 4% of waste
handlers were aware of risks involved in
biomedical waste handling. None has received
special training on this aspect, 5% suffered from
eye and skin infection and none reported to
higher authority. None was found immunized
against the infections (Table 3).
DISCUSSION
Biomedical waste seems to have received very
little attention in waste management process in
Srinagar city, Kashmir. Neither the government
nor hospital authority pays proper attention to its
management. Biomedical waste is disposed off
randomly leading to unhealthy and hazardous
environment affecting people living within the
vicinity of health institutions in particular and
city dwellers in general.
The study revealed that the hospitals do not
practice segregation, which was due to lack of
trained waste handlers and proper supervision.
As a result of failure to follow segregation
protocols and infrastructure, the waste as a
whole is potentially infectious. Rijal et al.,
(2007) observed segregation far from
satisfactory in most of the healthcare
institutions. Such a practice of non-segregation
may increase the costs of final disposal of the
waste.
Hospitals under study were found not to be
complying with the specifications for storage
facilities. Untreated waste in SKIMS was
transported to the collection point through
pipeline system, which consists of a network of
pipelines from various floors within the
hospital.The vertical conduit allows the waste to
be collected at a central collection point by
means of gravity, thus preventing the horizontal
movement of waste through the hospital
corridors. But, there are some hygienic and
19
technical problems associated with it (Bashir,
2009). Then the waste was carried to the onsite
incinerator plant. In SMHS hospital, waste was
collected manually and trolleys meant fo the
same were not used at all. Part of the waste was
taken to the CBWTF and part to the municipal
site at Achan Syedpora for its disposal. Barring
SKIMS, liquid waste from the other hospital was
flushed into the River Jhelum without any
treatment. Observation similar to the present
investigation have also of course been made by
the Purvi et al., (2006) in Gujarat, India.
Doctors outscored nurses and sanitary staff in
knowledge
regarding
biomedical
waste
management
and
rules
prescribed
therein.Doctors rated 94.3% in SKIMS, 96% in
SMHS
hospital
with
regard
to
knowledge.Sanitary staff exhibited poor
knowledge in the tune of 26.6%
and
25% respectively.This was indicative of the fact
that the sanitary staff was never given even a
capsule training with regard to biomedical waste
management.Continuous awareness regarding
biomedical waste management was needed to be
promoted.
It was found that the waste handlers were not in
receipt of special training on biomedical waste
handling which was prerequisite and necessary
to ensure an understanding of the risks that
wastes pose, to know how to manage waste etc.
Waste handlers were found suffering from
infections with no reporting to higher
authorities. Reportedly, very few waste handlers
were found immunized against the infections.
Hence implementation of a waste management
policy, training and motivation must be given
paramount importance to meet the current needs
and standards of biomedical waste management
(Sharma and chauhan, 2008).
CONCLUSION
The management of biomedical waste in the
study centres do not conform to the Biomedical
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
waste (Management and Handling) Rules, 1998.
It seemed that no significant action has been
taken by the hospital authorities in compliance
with the rules. The resistant attitude of hospital
staff, lack of technical knowhow, and lack of
skilled manpower could be responsible for the
non-compliance
of
biomedical
waste
management rules.
In order to achieve an effective and sustainable
biomedical waste management system in the
hospitals, the following suggestions were put
forward for consideration:
(i)
Segregation practices were needed to be
imposed within hospitals to separate
infectious waste, which will result in a
clean solid waste stream.
(ii)
Demonstrative programs should be
conducted for employees who are in
direct contact with the biomedical waste
in order to provide an understanding of
risks and importance of health and
safety measures during handling.
(iii) Periodic meetings of staff involved with
the waste management be conducted in
order to discuss problems and provide
suggestions.
(iv)
Strict enforcement of law will help in
improving the overall biomedical waste
management scenario in Srinagar city.
ACKNOWLEDGEMENT
We acknowledge with thanks the (i)
administrative staff of SKIMS and SMHS
hospital, Srinagar, India for granting
permission and cooperation during the study
period and (ii) Scientists of State Pollution
Control Board for their support and invaluable
help that provided this study much of its pace
and momentum. The noted patience of
respondents is also highly appreciated. Authors
acknowledge the immense help received from
the scholars whose articles are cited and
included in reference of this manuscript. The
20
authors
are
also
grateful
to
authors/editors/publishers of all those articles,
journals and books from where the literature for
this article has been reviewed and discussed.
1.
2.
3.
4.
5.
REFERENCES
Baram, M. 1989. Hospital management of
medical waste: legal framework and policy
issues, Ch. IV: Perspectives on Medical
Waste, Albany, NY: The Nelson A.
Rockfeller Institute of Government, State
University of New York.
Bashir, F. 2009. Study of solid waste
management at SKIMS. School of health
sciences IGNOU.
Bates, E.C. 2005. A Gazetteer of Kashmir.
Pp: 352.
Baveja, G., Muralidhar, S., Aggarwal, P.,
2000. Hospital Waste Management- an
overview. Hospital Today 5 (9), 485-486.
Belkin, N.L. 1982. Do reusable or disposal
gowns give better protection against
infection? Laundry News, 8 (2): 10, 17-18.
6. Campbell, D. 1988. Hospital Waste
Management in Canada, proceeding of the
national workshops on hospital waste
incineration and hospital sterilization, U.S.
EPA, San Francisco, C.A.
7. Manohar, D., Reddy, P.R., Kotaih, B., 1998.
Characterization of solid waste of a
superspeciality hospital – a case study. Ind. J.
Environ. Health 40 (4), 319-326.
8. Nugget, Hospital Waste Management and Biodegradable Waste, Government of India, Press
Information
Bureau,
http://pib.nic.in/infonug/infaug,99/i3008991.ht
ml-downloaded on 25.04.2010.
9. Pandit, N.A. 1999. Study of biomedical waste
management in teaching hospitals of Kashmir,
P.hd thesis, Dept. of Hospital Administration,
SKIMS.
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10. Pruss, A., Townend, W.K. 1998. Teacher‘s
Guide: Management of wastes from healthcare
activities. Geneva, WHO. Pp: 160.
11. Purvi, M., sheth, K.N., Desai, H. 2006.
Characterization
and
management
of
biomedical waste in SAE hospital, Anand- a
case
study,
Electronic
journal
of
environmental,
agricultural
and
food
chemistry, 5 (6): 1579-4377, 1583-89.
12. Rijal , K., Despande, A. 2007. Critical
evaluation of biomedical waste management
practices in Kathmandu valley. Proceedings of
the International Conference on Sustainable
Solid Waste Management. Pp: 142- 47.
13. Sharma, S., Chauhan, S.V.S. 2008. Assessment
of biomedical waste management in three apex
Government hospitals of Agra, 29(2): 159-162.
14. Stamenkovic, M., Kralj, D. 2007. Healthcare
and waste management, WSEAS Int.
Conference on energy planning, energy saving,
environmental education, Arcachon, France.
Pp: 116-19.
Table 1 Segregation of biomedical waste as per BMW Rules, 1998
Color Coding
Yellow
Red
Blue/White
Black
Waste Category
Cat. 1, Cat. 2 & Cat 3
Cat. 3, Cat. 6 & Cat 7
Cat. 4 & Cat 7
Cat. 5 & Cat 9
Table 2 Training of waste handlers and particulars regarding risk involved in waste handling at
SKIMS
S. No
Training and other particulars
No. of waste handlers
1.
Received special training in BMW handling
08
%
(n=110)
7.27
2.
Aware of risk involved in BMW handling
11
10
3.
An injury/ puncture /infection
20
18.18
4.
Accident reporting to higher authority
0
0
5.
Immunization
04
3.63
Table 3 Training of waste handlers and particulars regarding risk involved in waste handling at
SMHS Hospital
21
S. No
Training and other particulars
No. of waste handlers
%
(n=100)
1.
2.
3.
4.
5.
Received special training in BMW handling
Aware of risk involved in BMW handling
An injury/ puncture /infection
Accident reporting to higher authority
Immunization
0
4
5
0
0
0
4
5
0
0
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Fig.1. Flow diagram of sewage treatment plant, SKIMS.
Fig.2. Level of awareness among the hospital staff at SKIMS and SMHS Hospital
22
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
PATIENTS‟ PERCEPTION ON HOSPITAL SERVICES WITH
SPECIAL REFERENCE TO BEHAVIOR OF DOCTORS IN
VILLUPURAM DISTRICT
ijcrr
Vol 04 issue 08
Category: Research
Received on:15/02/12
Revised on:07/03/12
Accepted on:23/03/12
D. Karthikeyan, M. Thurunarayanasamy
Commerce Wing DDE, Annamalai University, Annamalai Nagar
E-mail of Corresponding Author: karthikeyan14390@gmail.com
ABSTRACT
This study aims to examine the patient perception of hospital services in villupuram district. A total of
600 respondents were selected in the study area in and out patients were included in the study to know
their perceptions towards the public and private health facilities. The major reason of choosing the public
health facility was inexpensiveness, infrastructure, and proximity of health facility. From the analysis of
the respondents regardless of their social status have expressed the same ‗poor‘ views, but there is a
significant difference in the degree of poor opinion across respondents categories by age, sex, education
and occupation. It is finally concluded that behavior of doctors is significantly better in private hospitals
compared to Government hospitals in villupuram district.
____________________________________________________________________________________
INTRODUCTION
Studies of patient satisfaction towards health
services, health personnel and resources
constitute important elements in the extent to
which health services meet consumers‘
expectations and needs. In recent years, patient‘s
opinion is increasingly considered to be a useful
component in the determination of care
outcomes. Users‘ perceptions are now
considered to be important source of information
in screening for problems and developing an
effective plan of action for quality improvement
in health care organization1. Quality of health
care is the degree of performance in relation to a
defined standard of interventions known to be
safe and that have the capacity to improve health
within available resources. It can also be defined
as meeting the health needs at the lowest cost
and within regulations2. Traditionally quality of
health care has been measured using
23
professional standards and neglecting the
importance of patient perception and opinions in
assessments of medical services, It has gained
greater prominence over the past 25 years In
any field, including medicine, customers‘
perception on any service providing paramount
importance and it is necessary for continual
service improvement.3
Statement of the Problem
Last three decades provision of health care was
dominated by Govt - run Hospitals. Owing to
population explosion and consequent pressure
on hospital infrastructure, Government hospitals
could not cater to the needs of ever-growing
patient population. With the result, patients
belonging to middle class and upper middle
class started switching to private health care
providers in getting quality medical service.
The newer diseases, life style diseases caused by
environmental degrade warrant a heavy demand
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
on health care services. Besides Government
hospitals, and private hospitals have begun
health care needs proliferating to cater to the
mounting health care space. It is also apparent
that there is a growing dissatisfaction among the
patient clients about the services provided by
health care service providers.
Studies of patients‘ perception towards health
personnel, health services and resources are
important to determine whether they meet
patients‘ expectations and needs and to judge
patient satisfaction. This information can be
used by hospital management in the
improvements of programs and the problems
identified by the patients. This will further
provide a detailed picture of the patients‘
experience at the hospital from which the
hospital management can direct and focus their
resources for better service in the future.. In this
context this study proposed patients‘ perception
doctors on hospital services with in Villupuram
District.
Objective of the study
1. To measure the patients‘ perception on
behavior of health personal in select hospitals
in the study area.
METHODOLOGY
Period of the study
The required primary data would be collected
from the selected respondents during three
months period, from May 2011 to July 2011.
Secondary data will be collected for ten years
period from 2002 to 2011
Sources of data
Primary data will be collected through
questionnaires as well as through personal.
Secondary data will be collected from published
books, journals, and other documents
Sampling Design
The survey is proposed to conduct only on the
target population of the selected hospitals. The
24
details regarding the selected hospitals were
obtained from the Deputy Direct of Health
services, Villupuram. There are 575 public
sector
health
establishments
namely
Government hospitals, Primary Health Centres
(PHCs) and Health Sub-Centres (HSCs) in the
district. There are and 143 private sector health
establishments are namely private hospitals,
nursing homes and clinics. Stratified random
sampling was adopted for the selection of
hospitals.
Sampling Technique for Selection of
Hospitals and respondents
With regard to the selection of hospitals more
than 50 bedded hospital from both private and
government hospital in the study area will be
selected, ten hospital from the government and
ten from the private sector by using stratified
random sampling technique. Thirty patients
from each such sample hospital will be selected
using convenience sampling technique. Thus,
the sample size of the patients of Government
hospitals amounts to 300 and 300 patients from
private hospitals. Thus, the total sample size of
the patients for this would be 600; it consists
both in and out patients
RESULTS AND DISCUSSION
The reliability analysis calculates the following:
item to total correlation, alpha if deleted and
overall Cronbach‘s alpha coefficient.
The
Cronbach‘s alpha coefficient is widely used
measure to find out the reliability and validity of
a scale items. A scale items with Cronbach‘s
alpha value of 0.70 and above is considered as
reliable and valid in the acceptable level.
George and Mallery (2003) provide the
following rules of thumb: ― > 0.90 – Excellent,
> 0.80 – Good, > 0.70 – Acceptable‖ (p. 231).
As a rule of thumb, the cutoff value for item to
total correlation is 0.30 and above, and alpha if
deleted value should be less than overall
Cronbach‘s alpha coefficient for any item to be
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
retained in the scale. However, if alpha if
deleted value is less than overall Cronbach‘s
alpha, and item to total correlation is a bit less
than 0.30 (>= 0.25), then the item can be
considered for retaining in the measurement
scale.
Next to reliability analysis, data pertaining to
behavior of doctors are exposed to principal
components of factor analysis with varimax
rotation to identify the primary behaviours of
doctors.
The perceived status of primary
behaviours are then compared across different
socio-economic categories of respondents. This
is carried to know whether the respondents
regardless of the difference in socio-economic
characteristics have perceived the behavior of
doctors and nurses in similar manner or not. If
there is similarity in the perception regardless of
the socio-economic characteristics then final
perceived status based on the entire sample
regarding the doctors‘ as well as nurses‘
behavior will be irrefutable and conclusive. The
results of the analysis are tabulated and
discussed in the remaining part of this chapter.
Item No
Table .1 Results of Reliability Test for Scale Items Measuring Behaviour of Doctors
Description of Scale Items
1
2
3
4
The level of communication between patient and doctors
Time spend by doctors with patient
Advice given by doctor about ways to avoid illness
Explanation given by doctors about the cause of disease
Explanation given as reason for different medical test to be
5
made
6 Care taken by doctor to check everything
7 Providing information about condition and treatment
The level of understanding on language and medical terms
8
used by doctors
9 Trust worthiness, reliability and honesty of doctors
Routine preliminary test taken prior to admission of the
10
patient
Cronbach‘s Alpha Reliability Coefficient
Item to
Total
Correlation
0.3657
0.5546
0.4579
0.4921
Alpha if
Deleted
0.7846
0.7631
0.7738
0.7696
0.3889
0.7819
0.4331
0.5373
0.7767
0.7637
0.4590
0.7737
0.6060
0.7548
0.3261
0.7890
0.7913
Source: Primary Data
From Table 1, which presents the reliability
analysis for items in the scale measuring
behavior of doctors, it understood that ‗item to
total correlation‘ for all items is above 0.30 and
alpha if deleted value is below the overall
Cronbach alpha coefficient of 0.7913. Hence,
all 10 items in the scale used in the present study
for measuring behavior of doctors towards
patients are reliable and valid.
Table 5.2 and 5.3 provides the results of factor
analysis of the items in the scale measuring
25
doctors‘ behaviors. The eigenvalue produced
by the factor analysis is nothing but variance
explained in (extracted from) the actual original
data by an underlying factor. The size of the
eigenvlaue determines how many factors are
extractable (valid) from the actual data.
According to Kaiser Rule, a factor with
eigenvalue of one or above is considered as a
valid factor.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
TABLE NO .2 Eigenvalues of Factors Underlying Behaviour of Doctors
Factors
Eigenvalue
% of Total Variance
1
2
3
4
5
6
7
8
9
10
3.54
1.46
1.02
0.85
0.73
0.64
0.61
0.51
0.39
0.25
35.36
14.58
10.19
8.47
7.34
6.43
6.10
5.14
3.89
2.49
Cumulative % of
Total Variance
35.36
49.93
60.13
68.60
75.94
82.38
88.47
93.61
97.51
100.00
Source: Primary Data
In Table 5.2, it can be seen that the eigenvalue
of first, second and third factors is above one,
explaining 35.56 per cent, 14.58 per cent and
10.19 per cent of the variance in the actual data.
All these factors together could posses 60.13 per
cent of the essence of actual data. This further
reveals that there are three factors underlying the
behaviours of doctors and these three factors can
be extracted for further analysis. To know the
characteristics of each one of the valid factors,
loadings of items in the scale with these factors
are used.
Item No
TABLE NO.3 Factor Loadings of Items with Extracted Factors Underlying Behaviour of Doctors
(After Varimax Rotation)
Description of Scale Items
Factor 1
Factor 2
Factor 3
7
Providing information about condition and treatment
0.8581
-0.0154
0.1670
2
Time spend by doctors with patient
0.8204
0.0363
0.1692
9
Trust worthiness, reliability and honesty of doctors
0.6042
0.2693
0.3322
0.6694
0.1701
-0.1276
-0.0190
0.8618
0.0517
1
10
The level of communication between patient and
doctors
Routine preliminary test taken prior to admission of
the patient
3
Advice given by doctor about ways to avoid illness
0.1152
0.8196
0.1682
6
Care taken by doctor to check everything
0.3400
0.4540
0.1869
0.0653
0.1150
0.8104
0.2432
0.1070
0.7134
5
8
26
Explanation given as reason for different medical test
to be made
The level of understanding on language and medical
terms used by doctors
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
4
Explanation given by doctors about the cause of
disease
Explained Variance
0.3247
0.2850
0.4981
2.5207
1.8295
1.6627
% of Total Variance
25.21
18.30
16.63
Cumulative % of Total Variance
25.21
43.50
60.13
Providing
informatio
n
about
condition
and
treatment
Factor Label
Preliminary
test prior to
admission &
Advising
patients
to
avoid illness
Giving reasons
for conducting
different
medical test
Source: Primary Data. Boldfaced are high factor loadings.
From Table 5.3, which provides the loadings of
items in the scale with each one of the extracted
factors, it is understood that the first factor is
highly loaded with items 7 (Providing
information about condition and treatment), 2
(Time spend by doctors with patient), 9 (Trust
worthiness, reliability and honesty of doctors)
and 1 (The level of communication between
patient and doctors), second factor is loaded
with items 10 (Routine preliminary test taken
prior to admission of the patient), 3 (Advice
given by doctor about ways to avoid illness) and
6 (Care taken by doctor to check everything),
and third factor has high loadings of items 5
(Explanation given as reason for different
medical test to be made), 8 (The level of
understanding on language and medical terms
used by doctors) and 4 (Explanation given by
doctors about the cause of disease). Further, the
loading of items 7 and 2 with first factor, items
10 and 3 with second and item 8 with third
factor is very high. Hence, based on the items
27
with highest loadings, the first, second and third
factors is identified as the factors possessing the
behaviours of the doctors in respect of
―Providing information about condition and
treatment‖, ―conducting Preliminary test prior to
admission & advising patients to avoid illness‖,
and ―Giving reasons for conducting different
medical test‖. The perceived status of above
these three primary behaviours of doctors is
evaluated based on the entire sample as well as
across sub-sample groups based on their socioeconomic characteristics.
The mean of the entire sample is compared with
‗3‘, the value for neutral range using one-sample
t-test to statistically identify the opinion range.
The independent sample t-test and one-way
ANOVA is used to compare the means of two
groups and more than two groups respectively.
Table 5.4 presents the mean opinion level of
entire sample about primary behaviours of
doctors in both private and government
hospitals.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
TABLE NO .4 Behvariour of Doctors Based on Entire Sample
Factors Underlying
Behaviour of Doctors
Mean
SD
t Value
Providing information about condition and treatment
3.13
0.89
3.69***
Preliminary test prior to admission & Advising
patients to avoid illness
3.05
0.87
1.40
Giving reasons for conducting different medical test
2.92
0.87
-2.16**
**Significant at 5% level; ***Significant at 1% level.
An observation of the table shows that the mean
level of opinion of the entire sample with
―providing information about condition and
treatment‖ (Mean = 3.13), is significantly higher
than the neutral level (t-value = 3.69, p < 0.01)
whereas the opinion of the total sample
regarding ―giving reasons for conducting
different medical test‖ (Mean = 2.92) is
significantly less than neutral level (t = -2.16, p
< 0.05). On the other hand, the all respondents
in the sample have expressed neutral opinion
about ―Preliminary test prior to admission &
Advicing patients to avoid illness‖ (Mean =
3.05, t value is insignificant). Hence, it is found
that behavior of doctors in providing
information about condition and treatment is
good, giving reasons for conducting different
medical test is poor and conducting routine
preliminary test prior to admission and advising
patients to avoid illness is neither poor nor good.
TABLE NO. 5 Comparison of Perceived Status of Doctors‟ Behaviours among Patient Groups by
Age
Factors Underlying
Behaviour of Doctors
Providing information
about condition and
treatment
Preliminary test prior to
admission & Advising
patients to avoid illness
Giving reasons for
conducting different
medical test
Age (in Years)
F Value
18-30
31-40
41-50
51-60
> 50
3.48
(0.82)
2.99
(1.09)
3.01
(0.93)
3.16
(0.69)
3.11
(0.84)
5.18***
3.19
(0.67)
3.07
(0.92)
2.93
(0.86)
3.15
(0.94)
3.00
(0.87)
1.87
3.03
(0.83)
3.03
(0.66)
2.66
(0.96)
3.20
(0.75)
2.83
(0.95)
7.65***
Figure in brackets are standard deviation; Degrees of freedom = 4, 595 for F values.
Table value for 4, 595 df @10 = 1.95, @5%= 2.35; @1% = 3.35
**Significant at 5% level; ***Significant at 1% level
As shown in Table 5.5, the mean scores across
age groups ranges from 2.99 to 3.45 for
―Providing information about condition and
28
treatment‖, 2.93 to 3.19 for ―Conducting routine
preliminary test prior to admission & Advising
patients to avoid illness‖ and from 2.66 to 3.20
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
―Giving reasons for conducting different
medical test‖. From significant F value of 5.18
(p < 0.01) and 7.65 (p < 0.01), it is understood
that the opinion of the respondents about
behavior of doctors regarding ―Providing
information about condition and treatment‖ and
―Giving reasons for conducting different
medical test‖ differ by age while their opinion
about ―Conducting routine preliminary test prior
to admission & Advising patients to avoid
illness‖ is independent of the age.
TABLE NO. 6 Comparison of Perceived Status of Doctors‟ Behaviours among
Patient Groups by Sex
Sex
Factors Underlying
Behaviour of Doctors
Providing information about condition and
treatment
Preliminary test prior to admission & Advising
patients to avoid illness
Giving reasons for conducting different medical
test
Male
3.10
(0.92)
3.11
(0.88)
2.95
(0.82)
Female
3.18
(0.85)
2.97
(0.84)
2.88
(0.94)
t Value
1.08
2.02**
1.01
Figure in brackets are standard deviation; Degrees of freedom = 598 for t values.
Table value for 598 df @10 = 1.64, @5%=1.96; @1% = 2.58. **Significant at 5% level
From the comparison of opinion about
behaviour of doctors between male and female
patients, results of which are presented in Table
5.6, it is evident that both male and female
regardless of the difference in sex have
perceived
similarly
about
―Providing
information about condition and treatment‖ and
―Giving reasons for conducting different
medical test‖. At the same time, regarding
doctors‘ behavior in respect of ―Conducting
routine preliminary test prior to admission &
Advising patients to avoid illness‖, the level of
opinion of female group is significantly less than
that of male counterparts (t-value = 2.02, p <
0.01).
TABLE NO.7 Comparison of Perceived Status of Doctors‟ Behaviours among Patient Groups by
Education
Factors Underlying
Behaviour of Doctors
Educational Status
F Value
Illiterates
School
level
Under
Graduate
Graduate
Providing information about
condition and treatment
2.77
(0.83)
3.19
(0.80)
3.14
(1.09)
3.31
(0.73)
9.60***
Preliminary test prior to admission
& Advising patients to avoid
illness
2.68
(0.83)
3.07
(0.89)
3.12
(0.86)
3.19
(0.82)
9.85***
Giving reasons for conducting
different medical test
2.58
(1.07)
3.02
(0.89)
2.99
(0.73)
3.01
(0.80)
7.94***
Figure in brackets are standard deviation; Degrees of freedom = 3, 596 for F values.
Table value for 3, 596 df @10 = 2.09, @5%=2.61; @1% = 3.81. ***Significant at 1% level
29
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
When the opinion of the respondents compared
across categories by education using one-way
ANOVA test, results of which are depicted in
Table 5.7, it is understood that there is a
significant difference in the mean level of
opinion across groups by educational status.
This is because, F values, 9.60, 9.85 and 7.94 for
the difference in group means are all significant
at 1 per cent level. However, mean scores are
much below 3 only for illiterates. Hence, it is
found that perceived status of doctors‘ behavior
among patients is poor according to illiterates
and differ significantly from other educational
groups in this regard.
Table 5.8 presents the results of test comparing
the mean perception between patient group in
urban and rural areas.
TABLE NO. 8 Comparison of Perceived Status of Doctors‟ Behaviours among
Patient Groups by Location
Location
Factors Underlying
Behaviour of Doctors
t Value
Urban
Rural
Providing information about condition and
treatment
3.21
(0.84)
3.08
(0.92)
1.76*
Preliminary test prior to admission & Advising
patients to avoid illness
3.06
(0.84)
3.05
(0.88)
0.15
Giving reasons for conducting different medical
test
3.02
(0.81)
2.85
(0.91)
2.25**
Figure in brackets are standard deviation; Degrees of freedom = 598 for t values.
Table value for 598 df @10 = 1.64, @5%=1.96; @1% = 2.58. *Significant at 10% evel;
significant at 5% level
From the results presented in the table, the mean
and this level of opinion is significantly than that
level of opinion is significantly higher for urban
of rural patient group. At the same time,
group regarding ―Providing information about
regarding ―Conducting routine preliminary test
condition and treatment‖ (Mean = 3.21 Vs 3.08
prior to admission & Advising patients to avoid
and t-value = 1.76, p < 0.10) and ―Giving
illness‖, both urban and rural group have
reasons for conducting different medical test‖
perceived as neither poor nor good.
(Mean = 3.02 Vs 2.85 and t-value = 2.25, p <
Table 5.9 provides mean opinion across
0.05)) compared to that of rural counterparts.
respondent
categories
with
different
That is, urban patients perceive ―Providing
occupational status about behaviours of doctors
information about condition and treatment‖ as
in hospital towards patients.
good and ―Giving reasons for conducting
different medical test‖ as neither good nor bad
30
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
TABLE NO.9 Comparison of Perceived Status of Doctors‟ Behaviours among Patient Groups by
Occupation
Salaried
Professional
Unemployed
Providing information
about condition and
treatment
Preliminary test prior to
admission & Advising
patients to avoid illness
Giving reasons for
conducting different
medical test
Business
Factors Underlying
Behaviour of Doctors
Agriculturist
Occupational Status
2.91
(0.82)
3.09
(1.10)
3.34
(0.84)
3.31
(0.73)
3.14
(0.85)
5.35***
2.86
(0.93)
3.07
(0.96)
3.23
(0.75)
3.01
(0.76)
3.08
(0.84)
3.71***
2.74
(0.96)
3.12
(0.71)
3.03
(0.75)
3.05
(0.96)
2.82
(0.93)
4.46***
F Value
Figure in brackets are standard deviation; Degrees of freedom = 5, 594 for F values.
Table value for 4, 595 df @10 = 1.95, @5%= 2.35; @1% = 3.35. ***Significant at 1% level
According to the table, the mean perception of
the agriculture group against all three factors and
that of unemployed against ―Giving reasons for
conducting different medical test‖ is below the
neutral level (below 3.0). The mean scores for
other occupational groups ranges between 3.01
(for professional group regarding ―preliminary
test prior to admission & Advising patients to
avoid illness‖) and 3.34 (for salaried in respect
of ―Providing information about condition and
treatment‖. Further, F values 5.35, 3.71 and
4.46 for the different in group means against all
three factors are significant at 1 per cent level.
So, it is found that there is a significant
difference in the perceived status of doctors‘
behavior in hospitals among patients with
different occupational status.
Regarding behavior of doctors in private and
government hospitals, the opinion of the patients
is compared and results of the comparative
analysis are reported in Table 5.10.
Table 10 Comparison of Perceived Status of Doctors‟ Behaviours among Patient Groups by Sector
Factors Underlying
Behaviour of Doctors
Sector
t Value
Private
Government
Providing information about condition and
treatment
3.41
(0.67)
2.86
(1.00)
7.84***
Preliminary test prior to admission &
Advising patients to avoid illness
3.31
(0.90)
2.79
(0.75)
7.79***
Giving reasons for conducting different
medical test
3.35
(0.64)
2.50
(0.87)
13.57***
Figure in brackets are standard deviation; Degrees of freedom = 598 for t values.
Table value for 598 df @1% = 2.58 .***Significant at 1% level
31
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
As reported in the table, the behavior of doctors
in government hospital is found to be poor as
mean perception of the patients, which ranges
between 2.50 and 2.86, is much less then neutral
level (value of 3) against all three factors. At the
same time, mean perception of the patient group
belong to private hospitals ranging from 3.31 to
3.40 is well above 3 (neutral level) and in ‗good‘
range. Moreover, the t-values, 7.84, 7.79 and
13.57 for the difference in mean opinion level
between private and government hospital patient
groups with regard to ―Providing information
about condition and treatment‖, ―Conducting
routine preliminary test prior to admission &
Advising patients to avoid illness‖ and ―Giving
reasons for conducting different medical test‖
are significant at 1 per cent level. Therefore, it
is concluded that doctors‘ behavior in
Government hospital is poor whereas it is good
in private hospitals.
CONCLUSION
In this study, an attempt was made to know
whether the respondents regardless of the
difference in socio-economic characteristics
have perceived the behavior of doctors. The
perceived status of primary behaviours is then
compared across different socio-economic
categories of respondents, based on the items
with highest loadings, the first, second and third
factors is identified which are ―Providing
information about condition and treatment‖,
―conducting Preliminary test prior to admission
32
& advising patients to avoid illness‖, and
―Giving reasons for conducting different
medical test‖. The perceived status of above
these three primary behaviours of doctors is
evaluated based on the entire sample as well as
across sub-sample groups based on their socioeconomic characteristics. From analysis of the
respondents regardless of their social status have
expressed the same ‗poor‘ views, but there is a
significant difference in the degree of poor
opinion across respondents categories by age,
sex, education and occupation. It is finally
concluded that behavior of doctors is
significantly better in private hospitals compared
to that of those in Government hospitals in the
villupuram district.
REFERENCES
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Barnard, K.,Putnam, S. M., & Inui, T. S.
(2000). The practice orientations of
physicians and patients: The effect of doctor–
patient congruence on satisfaction. Patient
Education and Counseling, 39(1), 49–59.
2. Bureau
ofIndian
Standard.
Quality
Management and Quality System Elements:
Guidelines for Services : IS : 14004 (part - 2),
1992.
3. Torres E. J. and Guo K. L., 2004, ―Quality
improvement techniques to improve patient
satisfaction‖ International Journal of Health
Care Quality Assurance Vol. 17, No. 6, pp.
334-338.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
DNA BARCODING, PHYLOGENETIC DIVERSITY STUDIES
OF
ETROPLUS
SURATENSIS
FISH
FROM
POORANANKUPPAM BRACKISH WATER, PUDUCHERRY
Sachithanandam V.1, Mohan P.M.1, Muruganandam N.2, Chaaithanya I.K.2,
Arun Kumar P3, Siva Sankar R3
ijcrr
Vol 04 issue 08
Category: Research
Received on:29/01/12
Revised on:16/02/12
Accepted on:03/03/12
1
Department of Ocean Studies and Marine Biology, Pondicherry University,
Andaman
2
Regional Medical Research Centre (ICMR), Andaman
3
Department of Ecology and Environmental Sciences, Pondicherry University,
Puducherry
E-mail of Corresponding Author: pmmtu@yahoo.com
ABSTRACT
Etroplus suratensis is known for the high commercial value fish available in South India. The
identification of the species of this fish cumbersome and inaccurate in different life stages of the fish.
Therefore, DNA sequence of cytochrome Oxidase subunit I gene was analysed for the species
identification and phylogenetic relationship of the species. The average genetic distance of conspecifics
species value was found to be 0.005%. The present work suggests that COI sequence provides sufficient
information on phylogenetic and evolutionary relationship to distinguish the Etroplus suratensis species,
the brackish waters species of pearl spots, unambiguously. Further, this work revealed that every species
having individual genetic distances depended upon the environmental stress and water quality, which play
an important role for its minor morphometric variations. Therefore, it was concluded that a DNA COI
barcoding tool can be used for fish identification by non technical personnel (other than taxonomist).
____________________________________________________________________________________
Keywords: DNA barcoding, COI, brackish
water, Pooranankuppam and Etroplus suratensis
INTRODUCTION
The chromids or the pearl-spots (Family:
Cichlidae) form an important group among the
brackish water fishes of the tropics. One of the
genus Etroplus contains E. suratensis fish is
inhabitant in fresh water and brackish water in
southern India. E. suratensis has many desirable
features which make them ideal fishes for
aquaculture like wide salinity tolerance, ability
to breed in confined waters, fast rate of growth,
good body weight, tasty flesh, highly adaptable
feeding
habits,
robust,
sturdy
body1.
Experimental cultures of this species show its
potential for polyculture and integrated farming
with poultry. In addition to export, it has high
demand in the local market and fetches a price
33
of more than US$ 3/kg2. These fish is available
throughout the year. The average production is
about 1000 kg/ha/year over 8-10 month growout period.
Morphometric studies are not only essential to
understand the taxonomy but also the health of a
species (including reproduction) in an
environment. The morphometric features of the
fish are unique to the species whereas the
variations in its feature are probably related to
the habit and habitat3.
Morphometric measurements have been widely
used to discriminate populations of various fish
species4-6. Fishes are considered to be
phenotypically more variable than most other
vertebrates, having relatively higher withinpopulation coefficients of variation of
phenotypic characters. Genetic polymorphism or
environmental
factors
may
induce
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
morphological variability among spatially
separated fish populations7, and phenotypic
plasticity in fish morphology has been
documented for various species, including
cichlids8,9. E. suratensis is known to have
variations in various morphological features
which are dependent on the geographical
partition. Further environmental comparisons of
these estuaries would be worthwhile in
understanding the evolution of such variations.
In addition, genetic investigations of the
variation and differentiation involving more
estuarine samples of E. suratensis will be useful
in substantiating the conclusions. The genotypic
and phenotypic variation of species is a prerequisite in conserving them. DNA barcoding is
highly efficient method in the analysis of genetic
divergences among species as well as for intra
species-level identifications10.
Among the marine living organisms of the IndoWest Pacific, Teleosts are among the bestdescribed, even though their systematics and
taxonomy still need considerable research
effort11,12. Southeast Asia has been identified as
one of the world‘s biodiversity hotspots based
on both plant and animal diversity13. Many time
taxonomic
ambiguities
exist
due
to
morphological and meristic similarities. Modern
taxonomic work includes analysis of a host of
other traits, including anatomy, physiology,
behaviour, genes, and geography, yet
morphological traits remain cornerstone14.
In such circumstances, DNA barcodes has
revealed that this could be helpful even for
larval stage fish taxonomical identification15. To
facilitate DNA barcode identification of fishes,
regional working groups are conjoining under
the Fish Barcode of Life (FISH-BOL)
initiative16, which seeks to establish a barcode
reference sequence library for all fishes17. The
phylogenetic systems, in combination with
conservation genetics, provide a critical frame
work for understanding diversity18 and predict
34
vulnerability to exploitation of tropical reef
fishes19.
As the morphometric measurements could lead
to misidentification of the species in different
life stages of fish especially E. suratensis, which
would affect the conservation strategy and the
market value of the same. Molecular taxonomy
appears to be the best tool for the species
identification and advantageous over the other
method of taxonomy, so the effort was taken to
identify the E. suratensis through molecular
taxonomy.
METHODS
Study Area, Sample collection and
preservation:
Fishes were collected from local fish landings at
Pondicherry brackish area of Pooranankuppam
(Fig.1). The identification of fishes was done as
described in FAO14. A piece of muscle in the
lateral line was collected and stored in 95%
ethyl alcohol at -20 C for DNA extraction. The
DNA Isolation and PCR condition work was
carried out in Department of Ocean studies and
Marine Biology centre at Port Blair, Andaman
and Nicobar Islands.
Molecular Taxonomy:
Total DNA extracted from 0.25g of muscle by
the standard proteinase-K/ phenol-chloroformisoamyl alcohol-ethanol method20. PCR
amplification of a 650bp DNA fragment coding
COI gene of the mitochondrial (mt) DNA
genome was amplified using published primer
set11. PCR components and conditions for 50 l
reaction were as described in our previous
work21. The PCR product was resolved in 1%
Agarose gel and visualized using Gel Doc
System, to confirm the presence of amplified
product sized 650 bp. Nucleotide sequencing
was performed using BigDye Terminator Cycle
Sequencing kit, following manufacturer‘s
instructions (Applied Bio-systems, Foster City,
CA, USA).
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Vol. 04 issue 08 April 2012
Sequence Analysis:
The DNA sequences of phenotypically identified
fishes were assembled using the SeqMan II
version 5.03 (DNASTAR). The sequence of
Etroplus suratensis reference sequences
retrieved from the NCBI GenBank were aligned
using Clustal W pair-wise and multiple
alignment of MEGA version 4.122. Sequence
divergence was calculated using the Kimura 2parameter (K2P) model23 and the mid-point
rooted Neighbour-joining (NJ) tree of K2P
distances was created to provide a graphic
representation of the species divergence24 (Fig.
2).
RESULTS
DNA barcoding is a unique concept with many
innovative attributes undertakes continuous
improvement in taxonomy. The estimated
Etroplus suratensis species DNA sequences
were submitted to GenBank under the
mentioned accession number in Table 1. NCBI
BLASTn result revealed that 21 reference
sequences were matched with maximum identity
of Etroplus suratensis of Puducherry as
described in Table 2. The average genetic
distance within the species (K2P) is 0.005.
The species genetic evolutionary pair wise
distance proximity was calculated by the species
similarity of genetic base pair. The Etroplus
suratensis DOSMB species closely related to the
species (FJ237544 – 0.006: GU5666028 –
0.006: FJ347966 – 0.006 and AY263870 –
0.009) in the Indian waters and USA
respectively (Table 3). The nucleotide
composition of Etroplus suratensis from the
present studied species is A = 23.4%, T =
29.9%, G = 18.6% and C = 28.1%. The average
nucleotide composition of E. suratensis among
the species level is noted as A = 23.48%, T =
30.08%, G = 18.1% and C = 28.36% (Table 4,
Fig.3).
35
The NJ and K2P genetic distances were created
to provide a graphic representation of the
patterns of divergences. Two distinct clad with
two sub clad of the same species were
recognized with more than 90% bootstrap value.
These two sub clad formation was identified
based on the independent assemblages of close
related species with differences in region and
environmental closeness. The results clearly
shows that every species having individual
genetic distances depended upon survival of any
species adaptation of the environmental stress
and its water quality, which play an important
role of significant values of the genetic distances
internally and morphometric minor variations
externally.
DISCUSSION
Fishes are largest group of vertebrates, which
exhibits remarkable diversity of morphological
attributes and biological adaptations25. In these
circumstances fish taxonomist facing a large
problems while the identification of fishes. To
overcome this problem, a morphology- based
identification combined with molecular based
approach for the species identification using
DNA barcoding would be an ideal tool26. This
tool is an efficient method for species-level
identification of the mitochondrial Cytochrome c
Oxidase I (COI) gene27. Mitochondrial DNA
(mtDNA) has been widely employed in
phylogenetic studies of animals because it
evolves much more rapidly than nuclear DNA,
resulting in the accumulation of differences
between closely related species 28-30.
This
study provides
the
interspecific
heterogeneity which enhanced the efficiency of
species identification through bar coding. This
was proved in Australian marine fishes11,
freshwater fish barcoding from Canadian31 and
carangid fishes from Indian waters32. The
variations of phenotypic character of species are
unique and it is probably related to the habit and
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Vol. 04 issue 08 April 2012
habitat among the variants of this species33.
Genetic variation of the green chromid has not
been studied previously of the genus E.
suratensis in Indian waters.
In the studies of bar coding, it has been reported
that K2P values between two species should be
greater than 0.0227,34 and in e.g. Indian mosquito
DNA barcoding average K2P values is 0.032935.
The present bar code exhibited K2P pair wise
genetic distances variation among the species
level is 0.005. However, such variation has
greater impact on the survival of the haplotypes
and its evolution.
The efficiency of species identification by
molecular method was enhanced by the
interspecific
heterogenetic
relationship
displayed36. Based on the above investigation it
is clearly evident that the E. suratensis identified
in Puducherry waters represented the haplotypes
species morphometrically, the other part of the
India and USA waters. However, bar code
results suggested that they are genetically varied.
Since, this species identified as haplotypes it
may be of low level differences in
morphometrically because of low genetic
distances. Results of phylogenetic and
evolutionary relationship of present study were
supported by earlier studies11. Further, it has also
confirmed that the DNA barcoding help to
recover phylogenetic information and to
understand the relationship with the species as
well as Order level36. The present study also
supported that mtDNA COI barcode region
offers best species identification, which is most
applicable and comparable with the mtDNA 12S
- 16S rRNA region sequences37.
CONCLUSION
The study concludes that the fish which
identified morphometrically and DNA barcoding
method using COI gene sequence are one and
same species of E. suratensis, the brackish
waters species of pearl spots. Further, this result
36
also informed that every species having
individual genetic distances depended upon the
environmental stress and water quality which
play an important role for its minor
morphometric variations. Moreover, the mtDNA
COI gene based identification provides high
resolution in species identification in fishes.
ACKNOWLEDGMENTS
The Authors express their sincere acknowledges
to Prof. J. A. K. Tareen, Vice-Chancellor of
Pondicherry University and the constant help
and encouragement of Dr. P. Vijayachari,
Director, Regional Medical Research Centre
(ICMR), Port Blair for the extension of facility
during this study. The manuscript valuable
review and commands supported by Chandal Lal
and Sayi Dev. We express thanks to the
Pondicherry University and Central Marine
Living Resources and Ecology (CMLRE) for
funding this work. Authors acknowledge the
immense help received from the scholars whose
articles are cited and included in references of
this manuscript. The authors are also grateful to
authors/ editors/ publishers of all those articles,
journals and books from where the literature for
this article has been reviewed and discussed.
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Vol. 04 issue 08 April 2012
Table 1 Etroplus suratensis species and their mtDNA COI Sequences GenBank accession Number
Sl No
1
Name of the species
Family:Cichlidae Etroplus suratensis
Common Name
Pearl spots - chromids
NCBI GenBank Number
JN228382
Table 2 BALSTn SEARCH REQUEST AND RESULTS COI PUBLIC RECORDS DATABASE:
Table 2a Identification Summary:
Taxonomic Level
phylum
class
order
family
genus
species
Taxon Assignment
Chordata
Actinopterygii
Perciformes
Cichlidae
Etroplus
Etroplus suratensis
Probability of Placement (%)
100
100
100
100
100
99.1
Table 2b A species level match has been made
Sl.
No‘s
Order
Family
Genus
Species
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Perciformes
Perciformes
Perciformes
Characiformes
Perciformes
Perciformes
Perciformes
Perciformes
Perciformes
Characiformes
Characiformes
Perciformes
Perciformes
Characiformes
Beloniformes
Perciformes
Beloniformes
Beloniformes
Beloniformes
Beloniformes
Cichlidae
Cichlidae
Cichlidae
Characidae
Haemulidae
Haemulidae
Haemulidae
Haemulidae
Haemulidae
Alestidae
Alestidae
Cichlidae
Cichlidae
Alestidae
Hemiramphidae
Lutjanidae
Hemiramphidae
Hemiramphidae
Hemiramphidae
Hemiramphidae
Etroplus
Etroplus
Etroplus
Pseudocorynopoma
Pomadasys
Pomadasys
Pomadasys
Pomadasys
Pomadasys
longipinnis
longipinnis
Paretroplus
Etroplus
suratensis
suratensis
suratensis
doriae
hasta
hasta
hasta
hasta
hasta
longipinnis
longipinnis
damii
maculatus
longipinnis
quoyi
johnii
limbatus
limbatus
limbatus
limbatus
39
Hyporhamphus
Lutjanus
Hyporhamphus
Hyporhamphus
Hyporhamphus
Hyporhamphus
Specimen
Similarity
(%)
99.07
98.92
98.77
85.23
82.33
82.33
82.33
82.33
82.33
82.31
82.27
82.25
82.22
82.19
82.18
82.18
82.18
82.18
82.18
82.18
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table 3 Kimura two parameter Genetic distances
Sl.No Species Name/ GenBank No‟s
1
Etroplus suratensis DOSMB
2
FJ237544-Etroplus suratensis
3
FJ347966 Etroplus suratensis
4
GU566028-Etroplus suratensis
5
AY263870 Etroplus suratensis
Average Interspecific K2P Distances is
K2P
0.000
0.006
0.006
0.006
0.007
0.005
Country
Pondicherry
India" 10.02 N 76.13 E
India" 10.02 N 76.13 E
Kerala, India"
USA
Table 4 Nucleotide composition:
Sl. No Species Names and GenBank nos.
1
Etroplus suratensis DOSMB
2
FJ237544 Etroplus suratensis
3
AY263870 Etroplus suratensis
4
FJ347966 Etroplus suratensis
5
GU566028 Etroplus suratensis
Average base pair composition
T%
29.9
30.4
30.2
30.5
29.4
30.08
C%
28.1
28.4
28.0
28.2
29.1
28.36
A%
23.4
23.4
23.6
23.4
23.6
23.48
G%
18.6
17.9
18.2
17.9
17.9
18.1
Fig 1 Study Area
40
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Vol. 04 issue 08 April 2012
Fig 2 Kimura two parameter (K2P) Genetic distances Values
Fig 3 Nucleotide composition of E. suratensis with references sequences species.
41
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
69
Etroplus suratensisDOSMB
AY263870-Etroplus suratensis
100
FJ237544-Etroplus suratensis
GU566028-Etroplus suratensis
FJ347966-Etroplus suratensis
DQ119195-Herotilapia multispinosa
100
100
HQ654752-Parachromis managuensis
HQ654751-Parachromis managuensis
GU817266-Australoheros facetus ssp
HQ956111-Scorpaeniformes sp.
54
FJ583406-Forcipiger flavissimus
FJ583412-Forcipiger flavissimus
100
FJ583407-Forcipiger flavissimus
74
FJ583411-Forcipiger flavissimus
70
HM882986-Hepsetus odoe
HM882988-Hepsetus odoe
81
HM882985-Hepsetus odoe
100
63
HM882987-Hepsetus odoe
HM882978-Hepsetus odoe
64
HM882979-Hepsetus odoe
HQ573341-Perciformes sp.
Fig 4 Neighbour-Joining (NJ) Method for Phylogenetic analysis and Evolutionary relationships of
E. suratensis with NCBI references sequence The bootstrap test (1000 replicates) is shown next to
the branches length is = 0.41765812 [Felsenstein 1985]. The evolutionary distances were computed
using the Maximum Composite Likelihood method. Codon positions included were 1st+2nd+3rd.
Phylogenetic analyses were conducted in MEGA4 [Tamura, et al 2007].
42
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
IS SKELETAL MUSCLE „A TARGET ORGAN‟ IN LONG
TERM UNCONTROLLED DIABETES MELLITUS? A
COMPARATIVE AND CORRELATIVE STUDY OF TYPE I
AND TYPE II DIABETES
Prathamesh Haridas Kamble1, Sunil Bhamre2
ijcrr
Vol 04 issue 08
Category: Research
Received on:07/02/12
Revised on:16/02/12
Accepted on:03/03/12
1
2
Dept. of Physiology; B.J.Medical College, Pune
Dept. of Microbiology; B.J.Medical College, Pune
E-mail of Corresponding Author: dr.prathamesh81@gmail.com
ABSTRACT
Background and Objective: Diabetes Mellitus is the most common endocrinal disorder worldwide.
Long term uncontrolled diabetes is associated with complications of eyes, kidney, heart, blood vessels
and nerves. Studies have been carried out to see the effect of diabetes on skeletal muscle strength but the
results are conflicting; while very few studies have considered the muscle endurance. Moreover, the
correlation of glycosylated haemoglobin levels (HbA1c) with handgrip strength (HGS) and hand grip
endurance (HGE) has not been studied. So the present study was carried out in 100 type I diabetics and
164 type II diabetics to compare the HGS and HGE with 100 and 160 normal healthy non diabetic
subjects respectively. Also the objective of this study was to determine the relation of HbA1c with HGS
and HGE. Research Methodology: HGS and HGE were measured using Handgrip dynamometer.
HbA1c was assessed by cation - exchange resin method using Monozyme‘s Glycohemin kit on
Transasia‘s semiautoanalyzer. Outcome of Study: Results of the study showed that type I & II diabetics
had significantly lower HGS than non diabetics. HGE was lower in type II diabetics while it was
significantly higher in type I diabetics as compared to controls. This study also indicated that HGS and
HGE had no significant correlation with HbA1c. Thus present study reveals that uncontrolled diabetics
are at a risk of decreased muscle strength and endurance and the magnitude of affection is highly
individual specific. Thus there is a need for development of strategies in the form of strict glucose control
and resistant training exercise program to slow or prevent rapid decline in muscle function in diabetics.
____________________________________________________________________________________
Keywords: Handgrip strength, Handgrip
endurance, glycosylated haemoglobin, diabetes.
INTRODUCTION
Diabetes mellitus is the most common endocrine
disorder. It is a syndrome of impaired
carbohydrate, fat and protein metabolism which
is characterized by hyperglycemia caused by
either reduced insulin secretion or decreased
sensitivity of tissues to insulin. The worldwide
43
prevalence rate of Diabetes Mellitus (DM) for
all ages was about 2.8 % in 2000 and projected
to be 4.4 % in 2030 [1]. The chronic
hyperglycemia and its associated metabolic
deregulation, is associated with potential long
term complications that can affect various
tissues like kidney, eye, heart, blood vessel and
nerve. [2] There is a new concept to explain
these long term complications of DM called as
‗hyperglycemic memory‘ which proposes that if
a cell remains in hyperglycemic environment for
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
certain duration, then it adapts to work in the
hyperglycemic state. [3] Meticulous control of
blood glucose can decrease the symptoms and
improve the diseased condition. Even after the
return of plasma glucose to normal or near
normal level, the progression of long term
diabetic complications still continues. [4] Thus,
measurement of glycosylated hemoglobin
(HbA1c), which provides the information about
the average blood glucose concentration over
preceding 6-8 weeks, is a good indicator of long
term complications of diabetes mellitus.
The possibility that the skeletal muscle is also a
target organ for diabetic complication was
suggested by Sayer A A et al who found reduced
muscle strength and impaired physical function
in Type 2 diabetes. [5] There have been many
studies of handgrip strength in diabetic patients
with conflicting results. Many reports have
suggested
possible
patho-physiological
mechanism also. Reduction in handgrip strength
is generally found in diabetics. [6, 7, 8] It seems
that reduction in handgrip strength has a linear
relationship with severity of diabetes which in
turn is in linear relationship with functional
ability of daily living activities. However, at the
present time there are no reports of functional
limitations in daily activities ascribable to
diabetes. The present study attempts to compare
handgrip strength and handgrip endurance in
type I, type II diabetics and normal subjects
(controls), to evaluate whether there is any
correlation between glycosylated haemoglobin
(HbA1c) and magnitude of reduction in hand
grip strength and endurance in type I and type II
diabetic patients.
MATERIAL AND METHODS
The present study was carried out in the diabetic
clinic in Indira Gandhi Government Medical
College and Mayo hospital, Nagpur. The
Institutional Ethics Committee approved the
study. The study was divided into two groups:
44
A) Group I, B) Group II.
A) Group I: Based on detailed history and
physical examination, subjects were further
divided into two sub-groups: (i) Type I diabetic
group: Comprised of 100 male subjects in the
age group of 31-45 years, having duration of
diabetes between 5-10 years, regularly visiting
the diabetic clinic and taking regular insulin
therapy, were selected. (ii) Control I group: For
comparison, a separate group of 100 healthy
subjects, with no history of diabetes or disorder
of defective sugar metabolism, was selected.
They belonged to the same age group and nearly
had the same height, built, socioeconomic status
and ethnic group, as that of type I diabetic group
subjects.
B) Group II: Similarly, on the basis of detailed
history and physical examination, subjects were
further divided into two sub-groups (i) Type II
diabetics group: 164 males, belonging to age
group of 41-55 years, having duration of
diabetes between 5-10 years, regularly visiting
the diabetic clinic and taking only oral anti
diabetic drugs regularly, were included. (ii)
Control II group: A group of 160 healthy non
diabetic male subjects in the age group of 41-55
years and having nearly the same height, weight,
built, ethnicity and socioeconomic status were
selected.
Subjects, who were left handed, involved in
regular handgrip exercise or constant method of
working with handgrip or suffering from asthma,
chronic obstructive pulmonary diseases,
congestive cardiac failure, Myasthenia gravis
and hypothyroidism, were excluded from the
study. Also factors that interfere with HbA1c
test results like diagnosed cases of
hyperbilirubinemia and chronic alcoholism were
excluded from the study.
After selection, written informed consent was
obtained from all the participants. Then
anthropometric measurements like standing
height and weight were taken. Early morning 5
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
cc fasting blood sample was obtained under all
aseptic precautions. Serum was separated and
fasting blood sugar and glycosylated
haemoglobin levels (HbA1c %) were estimated.
Blood sugar levels were assessed by glucose
oxidase biosensor method using glucometer and
glycosylated haemoglobin levels (HbA1c %)
were assessed by cation - exchange resin method
using Monozyme‘s Glycohemin kit on
Transasia‘s semiautoanalyzer.
Handgrip strength was determined by using
handgrip dynamometer. The use of this
instrument was illustrated to participants prior to
testing. Handgrip dynamometer was given in the
right hand of subjects in standing position and
arm by their side, not touching the body and
were asked to squeeze the dynamometer with as
much force as possible, taking care to squeeze
only once for each measurement. 3 trials were
performed with a pause of about 10- 20 seconds
between each trial to avoid the effect of fatigue.
Best amongst the 3 measurements was noted.
The handgrip endurance was also measured. The
subjects were asked to maintain 80% of their
handgrip strength for as long as they could and
time in seconds was recorded using a stop
watch.
STATISTICAL METHODS
The statistical analysis of observations was
carried out. Mean and standard deviation were
calculated and significance of difference was
tested statistically by the unpaired student‘s ―t
test‖ at P ≤ 0.05. Correlation coefficient (r) was
calculated and tested for statistical significance.
RESULTS
Table no. 1 shows the various parameters and
their mean values and standard deviations for
both type I diabetics and type II diabetics and
their respective control groups.
The mean age of Group 1 (type I DM and
control I) was 37.8 years and 37.9 years, while
45
for Group 2 (type II DM and control II) it was
48.1 and 46.4. There was no significant
difference in the age of Group 1 and Group 2.
Thus both the groups were age matched.
There was no significant difference in height,
weight, BSA and BMI; indicating that the
groups were homogenous in this respect.
Fasting and post meal blood sugar levels were
higher in type I and type II diabetics than
respective controls.
For HbA1c the normal reference value is < 6 %.
[4] It was observed that for both type I and type
II diabetics, HbA1c was on a statistically higher
side than controls indicating poor control of long
term blood sugar levels.
The handgrip strength (HGS) was significantly
lower in type I and type II diabetics as compared
to controls. Handgrip endurance (HGE) was
significantly higher in type I diabetic subjects as
compared to controls, while for type II diabetics,
HGE was lower than the controls.
Table II depicts the correlation of Handgrip
strength and Handgrip endurance with various
parameters. It indicated that there was no
statistically significant correlation existing
between HGS and HGE with any of the
parameters of present study for both Group I and
Group II diabetics. This shows that the
magnitude of skeletal muscle strength and
endurance changes produced due to uncontrolled
diabetes
were
based
upon
individual
susceptibility of subjects.
DISCUSSION
The present study has demonstrated that both
type I and type II diabetic subjects had lesser
muscle strength than non diabetics. This finding
is consistent with the findings of Park SW [6],
Savas S [7] and Lord SR [8].The probable
explanations for this finding are: (1) diabetes is
associated with increased systemic inflammatory
cytokines such as tumor necrosis factor α and
interleukin-6. These cytokines have a
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
detrimental effect on muscle function. [9,10, 11]
(2) Uncontrolled diabetics are associated with
glycation of skeletal muscle proteins such as
actin and myosin leading to a significant
reduction in vitro speed of actin and myosin
filament. [12] (3) Cotter M 1989 [13], Klueber
KM 1989 [14] and Medina -Sanchez M 1991
[15] demonstrated a significant and selective
atrophy of type II b fibers in diabetic rats
although this mechanism remains unclear in
human (4) As suggested by Anderson H 1996
[16] motor neuronal neuropathic processes give
rise to peripheral neuropathy which might be
associated with decreased muscle strength in
type I and type II diabetic subjects, and (5) long
term uncontrolled diabetes leads to metabolic
consequences like muscle protein catabolism
and inadequate energy use, which results in
potential reduction in muscle strength.
Handgrip endurance in the present study was
significantly longer in type I diabetics than non
diabetics, while for type II diabetics it was
significantly shorter than the controls. Though
the mechanism of this finding is unclear in
humans, in experimental diabetic rats it has been
demonstrated that prolonged increased or
decreased blood insulin levels lead to a change
in the composition of muscle fiber type.
Hypoinsulinemia shifts the muscle fiber
composition towards red muscle fiber also
known as fatigue resistant fibers [10, 11] and
hyperinsulinemia induces an increase in the
number of white muscle fibers which are least
fatigue resistant. Thus due to hypoinsulinemia
seen in type I diabetics, there is an increased
proportion of fatigue resistant red muscle fibers
which might be responsible for increased
handgrip endurance in them. While type II
diabetes is a condition characterized by insulin
resistance and high blood insulin levels. So this
prolonged
hyperinsulinemia
induced-easy
fatigable white muscle fiber composition of in
case of type II diabetics, might be the reason for
46
their lower endurance as compared to non
diabetics.
In the present study, there was no
linear correlation of glycosylated hemoglobin
(HbA1c %) with handgrip strength and handgrip
endurance for both type I and type II diabetics.
These findings suggest that in uncontrolled
diabetics skeletal muscle weakness is produced
but the magnitude of affection depends upon
individual subject's susceptibility to the
glycemic changes as well as the irregularities in
the treatment compliance of each subject. Thus
considering these two factors the linear
correlation might not have been observed.
CONCLUSION
From the present study it is clear that if the
blood sugar level in diabetics remains
uncontrolled then they are at risk of decreased
skeletal muscle strength and its magnitude of
affection is highly individual specific. It is
important because the accelerated loss of muscle
strength may lead to functional limitation and
physical disability and morbidity. We need to
develop strategies to slow or prevent rapid
declines in muscle function in high risk
population of adults with diabetes to decrease
morbidity. Every potential way such as strict
glucose control and resistive training exercise
programs should be introduced.
ACKNOWLEDGEMENTS
We acknowledge the immense help received
from the scholars whose articles are cited and
included in references of this manuscript. We
are also grateful to authors / editors / publishers
of all those articles, journals and books from
where the literature for this article has been
reviewed and discussed.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
REFERENCES
1. Rathmann W, Giani G. Global prevalence of
diabetes: estimates for the year 2000 and
projections for 2030. Diabetes Care. 2004;
27(10):2568-9.
9. Temelkova-Kurktschiev T, Henkel E,
Koehler C, Karrei K, Hanefeld M.
Subclinical inflammation in newly detected
Type II diabetes and impaired glucose
tolerance. Diabetologia. 2002; 45(1):151.
2. Alberti KG, Zimmet PZ. Definition,
diagnosis and classification of diabetes
mellitus and its complications. Part 1:
diagnosis and classification of diabetes
mellitus provisional report of a WHO
consultation. Diabet Med. 1998; 15(7):53953.
10. Visser M, Pahor M, Taaffe DR, Goodpaster
BH, Simonsick EM, Newman AB, et al.
Relationship of interleukin-6 and tumor
necrosis factor-alpha with muscle mass and
muscle strength in elderly men and women:
the Health ABC Study. J Gerontol A Biol
Sci Med Sci. 2002; 57(5):M326-32.
3. Wright A. Metabolic memory in type 1
diabetes. Br J Diabetes Vasc Dis. 1998;
9:254-257.
11. Helmersson J, Vessby B, Larsson A, Basu S.
Association of type 2 diabetes with
cyclooxygenase-mediated inflammation and
oxidative stress in an elderly population.
Circulation. 2004; 109(14):1729-34.
4. Foster DW. Diabetes Mellitus. In: Fauci AS,
Martin JB, Kasper DL, Hauser S. (eds),
Harrison‘s Principles of Internal Medicine.
2011. Volume 2, (pp.2078-2080.) New
York; McGraw Hill.
5. Sayer AA, Dennison EM, Syddall HE,
Gilbody HJ, Phillips DIW, Cooper C. Type
II Diabetes, muscle strength and impaired
physical function. Diabetic Care. 2005; 28
(10): 2541-2542.
6. Park SW, Goodpaster BH, Strotmeyer ES,
de Rekeneire N, Harris TB, Schwartz AV, et
al. Decreased muscle strength and quality in
older adults with type 2 diabetes: the health,
aging, and body composition study.
Diabetes. 2006; 55(6):1813-8.
12. Ramamurthy B, Hook P, Jones AD, Larsson
L. Changes in myosin structure and function
in response to glycation. FASEB J. 2001;
15(13):2415-22.
13. Cotter M, Cameron NE, Lean DR,
Robertson S. Effects of long-term
streptozotocin diabetes on the contractile
and histochemical properties of rat muscles.
Q J Exp Physiol. 1989; 74(1):65-74.
14. Klueber KM, Feczko JD, Schmidt G,
Watkins JB 3rd. Skeletal muscle in the
diabetic
mouse:
histochemical
and
morphometric analysis. Anat Rec. 1998;
225(1):41-5.
7. Savas S, Koroglu BK, Koyuncuoglu HR,
Uzar E, Celik H, Tamer NM. The effects of
the diabetes related soft tissue hand lesions
and the reduced hand strength on functional
disability of hand in type 2 diabetic patients.
Diabetes Res Clin Pract. 2007; 77(1):77-83.
15. Medina-Sanchez M, Rodriguez-Sanchez C,
Vega-Alvarez JA, Menedez-Pelaez A,
Perez-Casas A. Proximal skeletal muscle
alterations in streptozotocin-diabetic rats: a
histochemical and morphometric analysis.
Am J Anat. 1991; 191(1):48-56.
8. Lord SR, Caplan GA, Colagiuri R, Colagiuri
S, Ward JA. Sensori-motor function in older
persons with diabetes. Diabet Med. 1993;
10(7):614-8.
16. Andersen H, Poulsen PL, Mogensen CE,
Jakobsen J. Isokinetic muscle strength in
long-term IDDM patients in relation to
diabetic complications. Diabetes.1996;
45(4):440-5.
47
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table no. I- Various parameters and their mean values and standard deviations for type I and type
II diabetics and their respective control groups
Sr.
No
1
2
3
4
5
6
7
8
9
10
Variable
Type I DM
Age (years)
Height (cm)
Weight (Kg)
BSA (m2)
BMI (Kg/m2)
BSL –F(mg/dl)
BSL- PP(mg/dl)
HbA1c (%)
HGS (Kg)
HGE (Sec)
37.8 ± 1.7
162.4 ±4.4
59 ±10.7
1.61 ±0.13
22.3 ±4.05
203 ±69.4
322 ±75.9
9.3 ±1.5
39 ±2.9
11.2 ±1.9
Group 1
Control I
37.9 ±1.5
163.5±5.8
61.1 ±5.01
1.64 ±0.08
22.8 ±2.01
92.3±7.7
127.5 ±7.8
4.3 ±1
56.1 ±2.8
9.9 ±1.5
signific
ance
NS
NS
NS
NS
NS
S***
S***
S***
S***
S***
Type II DM
48.1 ± 1.5
163.3 ±6.6
58.5 ±6.3
1.62 ± 0.1
22 ±2.5
146 ±60.3
242.3 ±76.6
9.1 ±1.7
44.3 ±10.3
9.5 ±1.1
Group 2
Control II
46.4 ± 6.8
160.4 ±3.4
57.8 ±3.4
1.57 ± .06
22.9 ±0.95
91.8 ±5.3
118.2 ±4.4
4 ±0.6
55.7 ±2.3
10.7 ±4.6
signific
ance
NS
NS
NS
NS
NS
S***
S***
S***
S***
S***
Table no. II- Correlation between various parameters and Handgrip strength (HGS) and Handgrip
endurance (HGE) of type I and type II diabetics
Parameters
Age (years)
Duration of diabetes (years)
Height (cm)
Weight (Kg)
BSA (m2)
BMI (Kg/m2)
BSL – F (mg/dl)
BSL – PP (mg/dl)
HbA1c (%)
HGS (Kg)
HE (sec)
48
Hand grip Strength (Kg)
Type I DM
Type II DM
(r- value)
(r- value)
-0.03 NS
-0.03 NS
-0.23 NS
-0.023 NS
NS
-0.11
-0.11 NS
NS
0.44
0.44 NS
NS
0.27
0.27 NS
NS
0.48
0.48 NS
NS
-0.13
-0.13 NS
NS
-0.05
-0.05 NS
NS
-0.04
-0.04 NS
0.09 NS
0.09 NS
Hand grip Endurance (sec)
Type I DM
Type II DM
(r- value)
(r- value)
-0.34 NS
0.21 NS
-0.03 NS
0.12 NS
NS
0.006
0.17 NS
S**
0.44
-0.12 NS
NS
0.27
-0.016 NS
S**
0.48
-0.24 NS
NS
-0.19
0.05 NS
NS
-0.03
0.04 NS
NS
-0.19
-0.16 NS
NS
-0.68
0.09 NS
-
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
IN-SILICO STUDIES ON P43 PROTEIN FROM PLASMODIUM
FALCIPARUM
Tarun Kumar Bhatt
ijcrr
Vol 04 issue 08
Category: Research
Received on:20/03/12
Revised on:23/03/12
Accepted on:27/03/12
Department of Biotechnology, Central University of Rajasthan, Kishangarh
E-mail of Corresponding Author: tarunbhatt1982@gmail.com
ABSTRACT
Eukaryotic Aminoacyl-tRNA synthetases exist in large complex consists of different tRNA synthetases
with auxiliary proteins. P43 is one of the three non-synthetases proteins found in multi-synthetases
complex. P43 has been shown to involve in various biological processes like tRNA transport from
nucleus, apoptosis etc. Homologous sequence of P43 is also found in Plasmodium falciparum (PfP43). In
this study, homology modeling, structure validation and active site determination methods were used to
perform structural characterization of P43. Results show the overall three-dimensional structure of P43
with proper Ramachandran plot. Also, Active site residues were nicely located onto the structure of P43.
In addition, structural comparison between P43 of human and parasite origin provided information on
subtle differences in overall structures of proteins. Our results suggest that elucidation of PfP43 structure
is critical in developing anti-malarial drugs.
____________________________________________________________________________________
Keywords: P43, Homology modeling,
Plasmodium
INTRODUCTION
Plasmodium falciparum is the causative agent of
epidemic disease malaria. Many developing
countries are suffering from socio-economic
burden of this fatal parasitic disease. Several
drugs have been identified against malaria
parasite but prime concern remains are the rapid
development of drug resistance among parasites.
In addition, P. falciparum adapts different
strategies to overcome immune responses1-4 and
because of it, effective vaccine against parasite
has not been developed. Taking into
consideration all the facts discussed above, there
is regular need of identifying new protein
49
molecules of the parasite which could be
targeted as potential drug target.
Aminoacyl-tRNA synthetases (aaRSs) are
conserved class of proteins which play important
role in protein synthesis machinery of all living
organism. In eukaryotes, aaRSs are found in the
form of multi-synthetases complex (MSC),
comprises of 9-10 different tRNA synthetases
and 3 non-synthetases proteins5. P43 is part of
the non-synthetases component of MSC. Protein
P43 has been shown to involve in different
biological processes which include trafficking of
tRNA, involvement in autoimmune disease,
inhibition of formation of new vascular tissue in
metastatic carcinoma and in stability to MSC6-9.
In addition, the important function of P43 comes
into play as precursor of EMAPII (Endothelial
Monocyte Activating Polypeptide) domain.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
EMAPII is involved in acute inflammation and
play crucial role in apoptotic processes10. This
aaRSs family of proteins have been identified in
Plasmodium along with the homolog of P43
(PfP43). Nothing much has been done in
characterization of this protein but PfP43 was
found to be secretory in nature during parasite
asexual life cycle in human. Pro-inflammatory
property of PfP43 might play an important role
in modulating host immune response and could
be vital in malaria patho-physiology. In this
work, we have utilized the quick and effective
method of solving three-dimensional structure of
PfP43 using homology modeling. Comparative
studies with human counterparts along with the
identification of active site of the protein P43
could pave the way in identifying new effective
drug-like molecules against deadly malaria
parasite.
MATERIALS AND METHODS
The sequence of PfP43 was obtained using
NCBI Blast by taking human counterpart as
template. Other information of PfP43 was
extracted from PLASMODB using PF14_04013
as accession number. 1E7Z and 1FL0 pdb
structures were used as a template for homology
modeling. Identification of template structures
was carried out using NCBI BlastP where search
parameters were restricted to PDB (Protein Data
Bank). Sali‘s Modeller and Swiss Model Server
were used to build the in-silico structure of
PfP43. Online facility of sequence submission
and locally downloaded program of Modeller,
both were used to construct three-dimensional
structure of P43 domain. RAMPAGE online
server was used for structure validation which
gives output of Ramachandran plot describing
maximum allowed amino acids present in
modelled structure. Active site prediction was
performed with CASTp using modelled structure
of PfP43 domain. Images were created using
50
CHIMERA11. Images were processed at higher
resolution in PNG format.
RESULTS AND DISCUSSION
The three-dimensional structure of PfP43
domain is highly compact in nature and it is
typically EMAPII like domain. Structure is the
mixture of alpha helices and beta sheets where
beta sheets are predominantly occupying the
most of the space (fig.1). In addition there are
several loops hanging out of the core part of
structure probably involved in making contact
with interacting molecules which include both
protein and nucleic acid in case of P43. Panel B
of fig.1 shows the surface topology of PfP43
where most of the surface is positively charges
with intermittent negative charged patches,
indicative of nucleic acid binding ability of
PfP43. However, one side of the PfP43domain is
highly negative in nature typically nucleic acid
binding site whereas other side is mixture of
both negative and positively charged residues
which might be interacting with other proteins
based
on
charge
complementarily.
Ramachandran plot of the modelled PfP43
suggest that most of the amino acid residues are
in allowed region of three-dimensional space
and thus validate the homology modeling (fig.3).
Further, structural comparison between P43 of
human and Plasmodium was performed. Overall
the both the proteins share common fold and
domain topology but there are few structural
differences like secondary structure of beta sheet
present in PfP43 whereas absent in human
counterpart, subtle changes in three-dimensional
space of helices and loops (fig.2). These
structural differences could become basis of
drug development strategy as small differences
in three-dimensional space are enough for an
inhibitor to bind with variable affinity.
Computed Atlas of Surface Topography of
Proteins (CASTp) provided the predicted active
site location within PfP43. The amino acid
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
residues which make the active site pocket are
coloured in green and there 3D-space locations
are highlighted both in ribbon and surface
diagram (fig.4). The active site volume and area
are 149 Ao and 176.9, good enough to
accommodate one or two bases in case on
nucleic acid and two or three amino acids in case
of proteins.
CONCLUSION
To understand the mechanism of enzyme
reaction or binding of two protein molecules,
structural information play a Very critical role,
and to get the structure of proteins using X-ray
crystallography or NMR or Electron microscopy
is very expensive and time consuming process.
Thereby, we adopted relatively cheap and fast
method of solving three-dimensional structure
using molecular modeling. Homology modeling
of PfP43 provided the much needed structural
information required to understand involvement
of this protein in many biological processes. For
example, occurrence of highly negative patches
on one side of protein led us to speculate the
tRNA binding region which is necessary for the
function of transport of tRNA molecules out of
the nucleus as well as for the stable formation of
multi-synthetases complex. Note only that,
remaining area of protein in three-dimensional
space with variable charge distribution might be
responsible for binding to other cellular factors
engaged in apoptosis or inflammatory pathways.
In the end, structural differences between
PfP43and human counterparts might pave the
way for in-silico screening which might lead to
malaria specific drug like molecules discovery.
ACKNOWLEDGEMENT
I would like to thank Central University of
Rajasthan, Department of Biotechnology for
providing resources to conduct these studies. I
also thank Deepti Joshi for her help in
performing homology modeling. Authors
51
acknowledge the immense help received from
the scholars whose articles are cited and
included in references of this manuscript. The
authors are also grateful to authors / editors /
publishers of all those articles, journals and
books from where the literature for this article
has been reviewed and discussed.
1.
2.
3.
4.
5.
6.
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Schwartz O, Peglar M, Carucci DJ, Yates
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Udomsangpetch R. Plasmodium vivax
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plasmacytoid dendritic cells and the
induction of regulatory T cells. Eur. J.
Immunol. 2008, 38:2697–2705.
Kerjan P, Cerini C, Semeriva M, Mirande
M. The multienzyme complex containing
nine aminoacyl-tRNA synthetases is
ubiquitous from Drosophila to mammals.
Biochem Biophys Acta 1994, 1199:293-297.
Han JM, et al. Aminoacyl-tRNA synthetase
interacting multifunctional protein 1/p43
controls endoplasmic reticulum retention of
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Vol. 04 issue 08 April 2012
heat shock protein gp96: Its pathological
implications in lupus-like autoimmune
diseases. Am J Pathol 2007, 170:2042–2054.
7. Liu B, et al. Cell surface expression of an
endoplasmic reticulum resident heat shock
protein gp96 triggers MyD88-dependent
systemic autoimmune diseases. Proc Natl
Acad Sci USA 2003, 100:15824–15829.
8. Park SG, et al. Dose-dependent biphasic
activity of tRNA synthetase-associating
factor, p43, in angiogenesis. J Biol Chem
2002, 277:45243–45248.
9. Lee YS, et al. Antitumor activity of the
novel human cytokine AIMP1 in an in vivo
tumor model. Mol Cells 2006, 21:213–217.
10. Ko HS, et al. Accumulation of the authentic
parkin substrate aminoacyl-tRNA synthetase
cofactor,
p38/JTV-1,
leads
to
catecholaminergic cell death. J Neurosci
2005, 25:7968–7978.
11. Pettersen EF, Goddard TD, Huang CC,
Couch GS, Greenblatt DM, Meng EC and
Ferrin TE. UCSF Chimera - A Visualization
System for Exploratory Research and
Analysis, J Comput Chem. 2004, 25:16051612.
Figure 1: Three-dimensional structure of PfP43. Left panel showing modelled structure
in ribbon form whereas right panel display hydrophobicity surface in different directions
52
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Vol. 04 issue 08 April 2012
Figure 2: Structural comparison between human and
Plasmodium protein P43 domain AIMPII
Figure 3: Ramachandran plot of modelled P43 structure
53
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
242- S
292- L
342- F
392- I
R
N
H
T
L
K
N
C
N
Y
M
V
V
V
D
L
L
L
P
N
V
V
N
Q
G
L
V
G
Y
A
I
T
V
N
P
I
EQVE I H PDAD T LY C LK I NLG EDKP RD I C S G LRN K KNAEDL
LKEKSL RGKK SHGMVLCG S F DEKVE L LVP P NGV K I GER I L
DKNLS S DKEK NP F F H I QP HL I LKDGVAHYK DTKW I S SQGD
S
Figure 4: Predicted active site of P43. Upper panel showing the protein sequence of
modelled P43 structure where active site residues are labelled in green. Lower panel
shows the active site pocket of P43 in 3D space in ribbon and surface diagram.
54
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
COMPARATIVE
MICROBIOLOGIC
ANALYSIS
OF
SUBGINGIVAL PLAQUE SAMPLES IN TYPE II DIABETIC AND
NON
–
DIABETIC
PATIENTS
WITH
CHRONIC
PERIODONTITIS BY POLYMERASE CHAIN REACTION
ijcrr
Vol 04 issue 08
Category: Research
Received on:08/03/12
Revised on:16/03/12
Accepted on:25/03/12
Mythireyi D1, M G Krishnababa2, Kalaivani3
1
Dept of Periodontics, SRM Dental College, SRM University, Chennai
Dept of Periodontics, Sathyabama Dental College, Sathyabama University, Chennai
3
Dept of Periodontics, Tamilnadu Government Dental College and Hospital
Dr M G R University, Chennai
2
E-mail of Corresponding Author: vinayvela@rediffmail.com
ABSTRACT
Background: Although Immuno inflammatory relationship between periodontal diseases and diabetes
mellitus is acknowledged, the difference in putative periodontal microorganisms between diabetic and
non diabetic individuals is not well established. Aim: To compare the prevalence of two putative
periodontal pathogens namely Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis
in Type II Diabetic and Non Diabetic patients with chronic periodontitis by Polymerase chain reaction
Materials and Method: Sixty subjects were selected from the Department of Periodontics, Tamilnadu
Government Dental College and Hospital, Chennai – 03. 30 Type II diabetic patients with chronic
periodontitis were categorized as Group I and 30 Non diabetic patients with chronic periodontitis were
categorized as Group II based on American Dental Association classification 1997 and American
Academy of Periodontology classification 1999. Two sites- 1 healthy site and 1 diseased site were chosen
in each patient, Group I H, II H – healthy site samples and Group I D, II D- diseased sites samples.
Subgingival plaque was collected, DNA isolation was done & the presence of A.actinomycetemcomitans
& P.gingivalis DNA was determined by PCR. The PCR products were sequenced and confirmed. The
data was statistically analysed. Results: A.actinomycetemcomitans was detected in 6.7 %, 6.7%, 13.3%,
10% in Groups I H, II H, I D, II D respectively. P.gingivalis was detected in 40%, 46.7%, 46.6%, 53.3%
in Groups I H, II H, I D, II D respectively. When comparisons were made between Groups I H & II H and
Groups I D & II D for the two organisms, no statistically significant difference was obtained
Conclusion: The present study shows no statistically significant difference in the prevalence of
A.actinomycetemcomitans and P.gingivalis in Type II Diabetic and Non Diabetic patients with chronic
periodontitis.
____________________________________________________________________________________
Keywords:
Aggregatibacter
actinomycetemcomitans,
Porphorymonas
gingivalis, Diabetes, Periodontitis, Polymerase
chain reaction
INTRODUCTION
Chronic inflammatory periodontal disease
(periodontitis) is primarily an anaerobic Gram
55
negative oral infection that leads to gingival
inflammation, destruction of periodontal tissues,
loss of alveolar bone and eventual exfoliation of
teeth in severe cases 11. Certain organisms within
the microbial flora of dental plaque are the
major etiological agents of periodontitis.
Traditional thinking / paradigms have
maintained that periodontitis is an oral disease
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
and that the tissue destructive response remains
localized within the periodontium. Whereas
studies by Cohen DW et al 19704, Mattila KJ
et al 19898 have indicated that periodontitis may
produce a number of alterations in systemic
health.
Diabetes mellitus is a metabolic disorder
characterized by altered glucose tolerance or
impaired lipid and carbohydrate metabolism 1.
It has been suggested that a positive correlation
exists between diabetes and periodontal
destruction based on the fact that loss of
periodontal attachment occurs more frequently
and more extensively in moderately and poorly
controlled diabetic patients than those under
good control.
Diabetes mellitus influences prevalence and
severity of periodontal disease. Although host
immune inflammatory response plays an
important role, it is the microflora that‘s proved
to be the etiological agent in periodontitis
AIM
The aim of the present study was to compare the
prevalence of two putative periodontal
pathogens
namely
Aggregatibacter
actinomycetemcomitans and Porphyromonas
gingivalis in Type II Diabetic and Non Diabetic
patients with Chronic periodontitis by
Polymerase chain reaction.
MATERIALS AND METHOD
SUBJECT SELECTION
Sixty subjects were screened and selected from
the out patient Department of Periodontics,
Tamilnadu Government Dental College and
Hospital, Chennai – 600 003. 30 Type II
diabetic patients with chronic periodontitis were
categorized as Group I (Study group) and 30
Non diabetic patients with chronic periodontitis
patients were categorized as Group II (Control
group). Within each group sub categorization
was done as follows:
56
Group I H - 30 Healthy sites of Type II Diabetic
patients
with
chronic
periodontitis
Group I D - 30 Diseased sites of Type II
Diabetic patients with chronic periodontitis
Group II H - 30 Healthy sites of Non Diabetic
patients
with
chronic
periodontitis
Group II D - 30 Diseased sites of Non Diabetic
patients with chronic periodontitis
The criteria for selection of patients with Type 2
Diabetes was based on American Diabetes
Association classification (1997)1 and for
Chronic periodontitis and Chronic periodontitis
modified by Diabetes, the American Academy
of Periodontology classification (1999)2 was
utilized.
INCLUSION CRITERIA
Age 30 – 60 yrs, Either sex, At least 3 sites
with PPD 7 mm, CAL >1mm, At least 2
sites with PPD ≤ 3mm, Type 2 Diabetic
patients (Group I), Systemically healthy
individuals (Group II)
EXCLUSION CRITERIA
Patients with systemic disease other than
Type 2 diabetes(Group I), Patients with
systemic disease (Group II), Antibiotic
therapy for the past 6 months, Smokers,
Periodontal therapy for the past 1 year
Following Institutional Ethical Committee
Approval, selection of subjects was done.
Informed consent was obtained and a thorough
medical and dental history was taken. Intra-oral
examination was done using mouth mirror and
William‘s periodontal probe. Periodontal
evaluation was done by measuring the Plaque
Index, the Gingival Bleeding Index, Probing
Pocket depth( PPD) and Clinical Attachment
level(CAL).
Collection of subgingival plaque sample and
polymerase chain reaction
Two plaque samples were taken from the most
diseased site and a healthy site from each patient
with individual sterile Gracey curettes. The
plaque was dislodged into a vial containing
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
200 l of sterile lysis solution (10mm tris, 1.0
mm EDTA , 1.0% Tris X – 100, pH 78), sealed
and stored at -20oC. Subgingival plaque samples
was boiled for 10min, cooled to room
temperature, centrifuged at 10,000 rpm for 3 min
and the supernatant was stored at -20ºC till
assay. 10µl of the supernatant was directly used
as template in PCR.
Primers utilized in this study
Aggregatibacter actinomycetemcomitans (Aa)
Forward primer
:
5‘
CAGCAAGCTGCACAGTTGCAAA – 3‘
Reverse primer :
5‘
CATTAGTTAATGCCGGGCCG TCT – 3‘
(Kraig E et al / Infect Immun 1900, 58 : 920 –
929)
Porphyromonas gingivalis (Pg)
Forward primer
:
5‘
ATAATGGAGAACGCAGG AA -3
Reverse primer :
5‘
TCTTGCCAACCAGTTCCA TTGC – 3‘
(Dickinson et al / J Bacteriol 1998 ; 170 ; 1658
– 1665)
10 µl of the PCR master mixture was pipette
into micro centrifuge tubes. 1.0 l of forward
primer, 1.0 l of reverse primer, 3.0 l of
template DNA and 5.0 l of nuclease free water
was added and mixed thoroughly. The micro
centrifuge tubes were placed in thermocycler
and cycling conditions were set. PCR was
performed for 35 cycles of Denaturation at 95oC
for 1 min, Primer Annealing at 55oC for 30 sec,
Primary extension at 72oC for 1 min, Final
extension was 72oC for 10 min. The PCR
product was detected by 2% Agarose gel
electrophoresis. After the completion of the
electrophoresis, gel was taken to the
transilluminator and observed under UV-light.
(Biorad gel documentation)
The
PCR
product
of
238
bp
A.actinomycetemcomitans
and131
bp
P.gingivalis
were given to MWG –
BIOTECH, GERMANY for sequencing the
57
PCR products by automated DNA sequencer.
The data was collected and statistically
analyzed.
STATISTICAL ANALYSIS
Pearson‘s chi square test was used to calculate
the overall p value
The statistical package SPSS V18 (Statistical
Package for Social Science, Version 18) was
used for statistical analysis. In the present study,
< 0.05 was considered as the level of
significance.
RESULTS
In the present study A.actinomycetemcomintans
was detected in 6.7% Type II diabetic patients
with chronic periodontits -healthy sites (2 out of
30 healthy sites), 13.3% Type II diabetic patients
with chronic periodontits -diseased sites(4 out
of 30 diseased sites), 6.7% Non diabetic patients
with chronic periodontits - healthy sites (2 out
of 30 healthy sites) and10.0% Non diabetic
patients with chronic periodontits -diseased
sites(3 out of 30 diseased sites).
In the present study P.gingivalis was detected in
40% Type II diabetic patients with chronic
periodontits - healthy sites (12 out of 30 healthy
sites, 46.6% Type II diabetic patients with
chronic periodontits -diseased sites( 14 out of 30
diseased sites), 46.7% Non diabetic patients with
chronic periodontits - healthy sites(14 out of 30
healthy sites) and 53.3% Non diabetic patients
with chronic periodontits -diseased sites (16 out
of 30 diseased sites
DISCUSSION
The clinical parameters used in this study were
Gingival Bleeding Index, Plaque Index, Probing
Pocket Depth and Clinical Attachment level
similar to the study by Yuan K et al 200113.
Earlier studies employed curettes or paper points
for subgingival plaque collection. Sampling by
paper point is less invasive than by curette but
may result in an underestimation of tightly
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
adherent bacteria in subgingival sites5, 12.Hence
in this study curettes were used for sample
collection.
Although various diagnostic techniques are
available to analyse the microbial population of
subgingival plaque, PCR technique was opted as
it can detect organisms of less than 100 cells3.
In the present study, PCR procedure followed by
Yuan et al 200113 was used. Mandel et al 19907
detected 7% A.actinomycetemcomitans, 13% P.
gingivalis in diseased sites of NIDDM patients
by culture. Zambon et al 198813 employed
culture and ELISA assays and detected
P.gingivalis in 75% NIDDM subjects. In the
present study, 13.3% and 46.6% diseased sites in
Type II Diabetic patients with chronic
periodontitis and 10.0% and 53.3% of diseased
sites in Non Diabetic patients with chronic
periodontitis
were
positive
for
A.actinomycetemcomitans and P.gingivalis
respectively.
Our microbiological data revealed higher
detection rates when compared with the results
of others (except Zambon et al). This may be
due to the higher sensitivity of PCR, PCR is
more sensitive than culture, immunofluroesence
and DNA probes for which sensitivities are 2 x
102, 2 x 105, 2 x 104, 2 x 103 respectively6,14. In a
PCR study by Yuan et al 200113, 6.7% and
64.8% of diseased sites in Type II diabetic
subjects and 5.7% and 66.7% of diseased sites in
Non diabetic patients were positive for
A.actinomycetemcomitans and P.gingivalis
respectively. The discrepancy in results, may be
because a large sample size of 150 patients was
chosen by Yuan et al 200113 when compared to
the small sample of 30 patients chosen in this
study. Also, different authors have analyzed the
microorganism
distribution
in
different
population and races.
When comparing the prevalence rates of
A.actinomycetemcomitans and P.gingivalis, the
results in our study showed a lower detection
58
rate for A.actinomycetemcomitans which is in
concurrent to the study by Yuan K et al 200113.
This may be explained by the conclusion drawn
from the studies by Rhodenburg JP et al 19909 ,
Slots J et al 199010 that the prevalence of
A.actinomycetemcomitans was age related and
decreased
with
increasing
age.
A.actinomycetemcomintans is more responsible
for aggressive periodontitis whereas P.gingivalis
contributes to chronic periodontitis. Since our
subjects were all adults beyond 30 years of age,
it is speculated that the contribution of
A.actinomycetemcomitans to the periodontitis
we examined was minimal. There was no
statistically significant difference in the
detection rates of the 2 tested microorganisms
The plausible reasons are
1) There was no difference in the contribution
of the microbiological pathogens in patients
with Type II diabetic patients with chronic
periodontitis and in Non diabetic patients
with chronic periodontitis.
2) The PCR assay is limited in the ability to
differentiate large or small amounts of the
same pathogen.
3) Other microflora rather than our targeted
microorganism such as P.intermedia,
C.rectus and Capnocytophaga species
(since they are also regarded as important
pathogen in periodontitis of NIDDM
patients13 )could be an etiological agent in
Type II diabetic patients with chronic
periodontitis and Non diabetic patients with
chronic periodontitis.
CONCLUSION
The search for the etiologic agent of periodontal
diseases has been in progress for over a century.
In this study it was found that the composition
periodontal microflora in periodontal disease
sites of Type II diabetic patients with chronic
periodontitis was similar to that found in non
diabetic patients with chronic periodontitis.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
However, the significantly higher detection rate
of P.gingivalis in diseased sites further confirms
the possible pathogenic role of this bacteria for
both
groups
studied.
A.actinomycetemcomintans may not be a
causative pathogen in Type II diabetic patients
with chronic periodontitis and non diabetic
patients with chronic periodontitis.
Also, PCR assay provides only a binary results
i.e it detects the presence/absence of the
microorganism and cannot differentiate positive
result s quantitatively. Therefore the difference
may exist in the quantitative composition of
periodontal microorganism present in Type II
diabetic patients with chronic periodontitis and
non diabetic patients with chronic periodontitis.
Hence further studies with a large sample size
and diagnostic technique to quantitatively
analyse the composition of microorganisms may
be needed.
ACKNOWLEDGEMENT
Authors acknowledge the immense help
received from the scholars whose articles are
cited and included in references of this
manuscript. The authors are also grateful to
authors / editors / publishers of all those articles,
journals and books from where the literature for
this article has been reviewed and discussed.
REFERENCES
1. American Diabetes Association . Report of
the Expert Committee on the Diagnosis and
Classification of Diabetes mellitus Diabetes
Care 2001: 24 (Suppl 1 ) S5 –S20
2. Armitage GC: Periodontal diagnosis and
classification of periodontal diseases
Periodontology 2000 :2004; 34;9 -21
3. Ashimoto A, I Bakker I Slots : Polymerase
chain reaction detection of 8 putative
periodontal pathogens in subgingival plaque
of gingivitis and advanced periodontitis
59
4.
5.
6.
7.
8.
9.
10.
11.
lesions. Oral Microbiol Immunol 1996;
11;266 -273
Cohen DW, Friedman LA, Shapiro J, Kyle
GC, Franklin S Diabetes mellitus and
periodontal disease Two year longitudinal
observations Part I J Periodontol 1970 , 41;
709 -712
Hartroth B, Jeyfabrt I, Comado G Sampling
of periodontal pathogens by paper points :
Evaluation of basic parameters Oral
Microbiol Immunol 1999; 14; 326 -330
Maiden MFJ, Tanner A, Mc Ardle S,
Nagpauer K, Goodson JM Tetracycline
fibre therapy monitored by DNA probe and
culture methods J Periodont Res 1991
;26;452-459
Mandell RL, Dirienzo T, Kent R, Joshipura
K,Haber J Microbiology of healthy and
diseased periodontitis sites in poorly
controlled insulin dependant diabetes J
Periodontol 1992;63; 274- 279
Mattila KJ, Nieminen MS, Valtonen W,
Rasi VP, Kesaniemi YA, Syrjala SL
Association between dental health and
myocardial infarction BMJ 1989 ;298;779782
Rodenburg JP, Van Winkelhoff AJ, Winkel
EG, Goene RJ, Abbas F, de Graff J
Occurrence of Bacteroides gingivalis,
Bacteroides intermedius and Actinobacillus
actinomycetemcomitans
in
severe
periodontitis in relation to age and treatment
history J Clin Periodontol 1990; 17; 392-399
Slots J, Feik D, Rams JE Actinobacillus
actinomycetemcomitans and Bacteroides
intermedius in human periodontitis Age
relationship and mutual association J Clin
Periodontol 1990; 17;659 -662
Socransky SS and Haffajee AD
The
bacterial etiology of destructive periodontal
disease current concepts J Periodontol
1992;63;4;322-331
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Vol. 04 issue 08 April 2012
12. TannerAC, Goodson JM Sampling of
microorganisms associated with periodontal
disease Oral Microbiol Immunol 1986; 1; 15
-22
13. Yuan K, Chang CJ, Hsu pc, Sun HS, Tseng
CC, Wang Jr Detection of putative
periodontal pathogens in non insulin
dependant diabetes mellitus and diabetes
mellitus by Polymerase chain reaction
Periodont Res 2001; 36;18-24
14. Zappa u, Reinking – Zappa M ,Graf H,
Comparison of serological and DNA probe
analysis for detection of suspected
periodontal pathogens in subgingival plaque
samples Arch Oral Biol 1990; 35;161-164.
Table 1: Comparison of Distribution of Aa in Healthy Sites among Type II Diabetic and Non
Diabetic patients with Chronic periodontitis
Aa
P value
Diabetic Healthy n = 30
Present
Absent
Count
%
Count
2
6.7%
28
0.64 ( not significant )
%
93.3%
Non Diabetic Healthy n = 30
Present
Absent
Count
%
Count
2
6.7%
28
%
93.3%
Table 2: Comparison of Distribution of Pg in Healthy Sites among Type II Diabetic and Non
Diabetic patients with Chronic periodontitis
Pg
P value
Diabetic Healthy n = 30
Present
Absent
Count
%
Count
12
40%
18
0.301 ( not significant )
%
16%
Non Diabetic Healthy n = 30
Present
Absent
Count
%
Count
14
46.7%
16
%
53.3%
Table 3: Comparison of Distribution of Aa in Diseased Sites among Type II diabetic and Non
Diabetic patients with Chronic periodontitis
Aa
P value
Diabetic Diseased n = 30
Present
Absent
Count
%
Count
14
13.3%
26
0.12 ( not significant )
%
86.7%
Non Diabetic Diseased n = 30
Present
Absent
Count
%
Count
3
10%
27
%
90%
Table 4: Comparison of distribution of Pg in diseased sites among Type II Diabetic and Non
Diabetic patients with Chronic periodontitis
Pg
P value
60
Diabetic Diseased n = 30
Present
Absent
Count
%
Count
14
46.6%
16
0.24 ( not significant )
%
53.3%
Non Diabetic Diseased n = 30
Present
Absent
Count
%
Count
16
53.3%
14
%
46.6%
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Fig.1 : Distribution of Aa among
Groups I H, II H
6.70%
Fig.2 : Distribution of Pg among
Groups I H, II H
6.70%
46.70%
7%
48%
6%
46%
5%
44%
4%
42%
40.00%
3%
40%
2%
38%
1%
36%
0%
Diabetic Healthy
Diabetic Healthy
Non Diabetic Healthy
Fig.3 : Distribution of Aa among
Groups I D, II D
Fig.4 : Distribution of Pg among
Groups I D, II D
13.30%
53.30%
14%
54%
10.00%
12%
52%
10%
50%
46.60%
8%
48%
6%
46%
4%
44%
2%
42%
0%
Diabetic Diseased
61
Non Diabetic Healthy
Non Diabetic Diseased
Diabetic Diseased
Non Diabetic Diseased
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
238 bp
Photograph No.1: Electrophoresis showing the amplified product of A.actinomycetemcomitansM –
100 bp Ladder Lane 1, 2 – Positive for A.actinomycetemcomitans (238 bp) Lane 3, 4 – Negative for
A.actinomycetemcomitans Lane 5 Blank Control
131 bp
Photograph No.2: Electrophoresis showing the amplified product of P.gingivalis M – 100 bp Ladder
,Lane 1, 2, 3, 4 – Positive for P.gingivalis (131 bp)
62
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
EXPERIMENTAL BEHAVIOUR OF A PYRAMID TYPE
SOLAR STILL COUPLED AND DECOUPLED TO AN
ELECTRICAL TEMPERATURE CONTROLLER
S.Kalaivani1, S.Rugmini Radhakrishnan2, B.Selvakumar3, M.Indhumathy1
1
ijcrr
Vol 04 issue 08
Category: Research
Received on:08/02/12
Revised on:20/02/12
Accepted on:11/03/12
Department of physics, Vivekanandha College of Arts and Sciences for Women,
Tiruchengode, Tamilnadu
2
Department of physics, Avinashilingam University for Women, Coimbatore,
Tamilnadu
3
Department of physics, Jansons Institute of Technology, Karumathampatti,
Coimbatore, Tamilnadu
E-mail of Corresponding Author: drskvani@gmail.com
ABSTRACT
In this communication, an experimental investigation of the behaviour of a pyramid type solar still
coupled and decoupled to an electrical temperature controller unit has been presented. The main
advantage of the pyramid type solar still is that the maximum radiation can penetrate inside the basin
from electrical temperature controller. The main objective oh this present paper is to study the behaviour
of the still performance with and without of electrical temperature by analyzing the internal and external
heat transfer co-efficient.In general the still performance is reasonable with a good daily output including
nocturnal output. The addition of electrical temperature controller where capable of enhancing the
productivity with heat retention causing continued evaporation
____________________________________________________________________________________
Key words: Solar still, water collection, and
temperature controller.
INTRODUCTION
Mahmoud I.M. Shatat and K. Mahkamov (2010)
described the performance of a multi stage water
desalination still connected to a heat pipe
evacuated tube solar collector with aperture area
of 1.7 m². The multi stage solar still water
desalination system was designed to recover
latent heat from evaporation and condensation
processes in four stages.
V.K. Dwivedi et al., (2008) has made an attempt
to evaluate the internal heat transfer coefficients
of single and double slope passive solar stills in
summer as well as winter climatic conditions for
three different water depths (0.01, 0.02, and 0.03
m) by various thermal modes. The experimental
validation of distillate yield using different
thermal models was carried out for composite
63
climate of New Delhi, India (latitude 28º35‘ N,
longitude 77 º12‘E). By comparing theoretical
values of hourly yield with experimental data it
has been observed that Dunkle‘s model gives
better agreement between theoretical and
experimental results. Dunkle‘s model has been
used to evaluate the internal heat transfer
coefficient for both single and double slope
passive solar stills. With the increase in water
depth from 0.01 m to 0.03 m there was\a
marginal variations in the values of convective
heat transfer coefficients. It was also observed
that on annual basis output of a single slope
solar still is better as compared with double
slope solar still.
M.K. Phadatare et al., (2007) made an attempt
to study the effect of water depth on the internal
heat and mass transfer in a single basin single
slope plastic solar still. The experimental still
was fabricated from Plexiglas. The bottom and
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
all sided of the still are made from a sheet of
black Plexiglas. The cover is made from a
transparent Plexiglas of the same thickness. The
solar still was sealed to reduce the leakage of
vapour to the surroundings. The study covers the
influence of different environmental and
operational parameters on the still productivity.
The operational parameter such as depth of
water in the basin is varied from 2 cm to 12 cm
to find out its influence on internal heat and
mass transfer and hence the productivity of the
still. The maximum distillate output of 2.1
L/m2/day was obtained with water depth in still
basin 2 cm. The maximum efficiency of the
experimental still varied from 10% to 34%. The
results indicated that with increase in depth of
basin water, still productivity decreases
Salah Abdallah et al., (2007) developed an
experiment work to improve the single slope
solar still performance through increasing the
production rate of distilled water. Design
modifications were introduced to the
conventional solar still, involving the installation
of reflecting mirrors on all interior sides,
replacing the flat basin by step-wise basin, and
by coupling the conventional solar still with a
sun tracking system. The inclusion of internal
mirrors improved the system thermal
performance up to 30%, while step-wise basin
enhanced the performance up to 180% and
finally the coupling of the step-wise basin with
sun tracking system gave the highest thermal
performance with an average of 380%.
The accumulated production of the deep-basin
solar still and that of the tilted tray solar still
with longitudinal baffles are compared by
Badawi W.Tleimat et al., (2003). The
comparisons show that evaporation in the deep
basin still continues during the entire 24 hour
period while the tilted tray still ceases to
produce a relatively short time after sunset. Thus
nocturnal production is maximum for the deep
basin type and nearly zero for the tilted tray still
64
is studied. A small experimental still was
constructed to determine the factors affecting the
nocturnal production of solar stills. The
experimental results indicate that a substantial
increase of product water could be obtained
from the continuous addition of warm water to
the still. This increase was found to be a
function of flow rate, feed-water temperature,
evaporating and condensing areas and ambient
temperature.
Nikola Nijegorodor et al., (2003) described two
solar thermal –electrical methods to purify water
by distillation. In this first method air saturated
with water vapour is removed from a basin type
still by using a low power exhaust fan, and is
passed through a condenser where the latent heat
of water vapour is used to pre heat the
mineralized water for the basin. This also results
in lower temperature for the glazing and a faster
rate of evaporation from the basin. The net effect
in an increased thermal efficiency of the still
more than twice the thermal efficiency of the
conventional still. In the second design a
condenser collector is used to boil water in the
absorber tube. A low power vacuum pump is
employed to lower the boiling temperature of
water by about 10ºc. The yield of distillate from
the still is nearly double.
M.Boukar and A. Harmim (2001) has studied the
effect of desert climatic conditions on the
performance of a simple basin solar still and a
similar one coupled to a flat plate collector. A
three months round study showed that the
productivity of the simple basin and similar
coupled to a flat plate solar collector strongly
depends on the solar radiation and ambient
temperature
The effect of intermittent flow of waste hot
water in the basin of solar stills on its
performance has been discussed by Madhuri et
al., (2001) as well as the effect of various
parameters- duration of flow of hot water, flow
rate and water depth. It is concluded that for
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
higher yield, the waste hot water should be fed
into the basin during off-sunshine hours. The
results have also been compared with those of
Tiwari and Malik and sodha et al.
A single basin solar still with basin area of
0.98*0.98 m was constructed from galvanized
iron sheets and an inclined glass cover. The still
was provided with 525 W electrical heating
tapes, fixed under the still for indoor steady state
operation. The variations of basin temperature
and evaporation rate were measured during both
indoor and outdoor operation. Transient analysis
of the still requires the evaluation of
evaporative, convective and radiative heat
transfer coefficients. The Dunkle model, which
has been widely used for the prediction of the
evaporative coefficient, was found to be over
predicting evaporation rates. The models
developed in this work were found to provide
better prediction for the evaporation rate
measured in this work and this work was
investigated by Ahmad Taleb Shawaqfeh et al.,
(2000).
A techno-economic analysis of multi-stage
stacked tray (MSST) solar still coupled with a
solar collector through a heat exchanger has
been developed by R.S. Adhikari et al., (2000).
The study also includes a discussion on the
sensitivity of cost of unit mass of distilled water
in references to the useful life of distillation
chamber, cost of solar collector and other
associated parameters.
Faten H. Fahmy et al., (1998) has presented a
new way to realize continuous operation of a
solar desalination system to produce fresh water
using solar energy for a dual purpose. To realize
the continuity of still operation daily and
overnight, the batteries are discharged during the
night at a suitable rate to feed an electric heater.
The electric heater is designed to generate the
required heat for desalination during the night.
This modified still is provided with a packed bed
layer installed in the bottom of the basin to assist
the system during the day and at night, i.e., this
65
modified still will be more efficient. The use of
PV and packed bed systems means higher
efficiency than the passive still, as the modified
still produces large quantities of fresh water in
August for a saline water depth of 0.01 m by
using glass wool insulation 0.05 m thick and
glass spheres as a packed bed with 0.0213 m bed
length.
Multiple linear regression equations relating to
ambient, air temperature, wind speed and solar
radiation were developed by A.N. Minasian et
al., (1992) to estimate the productivity of a still.
The study shows that condensation process
inside the stills is achieved during the period
between sunset and sunrise. Results reveal that
the average wall‘s contribution in supplying
fresh water is about 56%, whereas base
contribution is about 31%. It is concluded,
therefore, that setting many stills on a number of
separated holes will give higher output rather
than setting a single still on one large hole of the
same volume.
H.M. Ali (1991) has been studied a mathematical
model to predict the performance of the solar
still using forced convection inside the solar still
to enhance the productivity of the still. It shows
that the productivity increases about 60% more
than that of a natural convection solar still. Good
agreement between the theoretical and
experimental results is obtained. Enhancing
mass transfer co-efficient due to forced
convection has a major role in the productivity
enhancements.
S.A. Lawrence and G.N.Tiwari (1991) have
been studied the thermal modeling based on heat
and mass transfer relations of a green house
integrated with a solar still. An experiment was
carried out for a typical greenhouse in Port
Moresby. The following observations were
made. (i) There is a reasonable agreement
between theoretical and experimental results and
(ii) the amount of distilled water obtained is
sufficient to grow the plants‘ inside the
greenhouse.
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Vol. 04 issue 08 April 2012
2. Construction details for both pyramid solar
still and temperature controller
Pyramid solar still of base area 0.85m x 0.85m is
designed. The still is filled with the water to a
height of 0.05m. Top of the system is covered by
a 3 mm transparent glass pyramid cover with a
height of 0.30m at the middle. It is air tightened
using the cushion supports at the interface
between the top cover and the sides of sliding
support for uniform landing. Bottom of the still
is insulated using sawdust, while the side is
insulated with glass wool. The specification of
different parts of the still is given below.
Fig (1) Cross sectional view of pyramid solar still
1. Top cover
4.
Water storage basin
7.
Still inlet
2. Water collection segment 5.
Still outlet
8.
Wooden box
3. Glass Wool Insulation
6.
Sawdust insulation
2.1
CONSTRUCTION
OF
ELECTRICAL
TEMPERATURE
CONTROLLER
Electrical backup made up of Temperature
controller, Heater coil and Thermocouple.
Temperature controller has various plug-in
from number 1 to number 20. For
temperature controller setting plug-in 1, 2,
N1, C, N0, 12 and 13 are used. Temperature
of the controller unit can be kept at any
temperature by using set key function (4
digit display), which has two buttons to
increase and decrease the temperature.
Connections from plug-in 2 and C are
66
interconnected and used as one end of input
power supply. Pug-in 1 is used as another
end of power supply input. Also a
connection is taken from Plug-in 1 and it is
connected to one end of the coil. Plug-in N0
is connected to another end of coil. Pluginn‘s 12 and 13 are connected to
thermocouples. Heater coil is made up of
stainless steel and is placed inside the solar
still. Sensor thermocouple should be placed
near to the heater coil. Schematic
representation of electrical backup circuit
with temperature controller is shown in Fig
(3.2).
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Vol. 04 issue 08 April 2012
Fig (2) Electrical temperature controller
Since controller switch is connected to input
power supply, once when the fixed temperature
is attained, the circuit collapses. As a result
power supply is shut down. When the
Fig. (3) Experimental Set Up Of The
Pyramid Cover Solar still
temperature falls below the set value, circuits is
switched on and the current flows through the
coil, hence it starts heating.
Fig. (4) Electrical Temperature Controller
Fig. (5) Experimental Set Up Of The Pyramid Cover Solar Still
Coupled With Electrical Temperature Controller
67
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Vol. 04 issue 08 April 2012
3.Thermal Analysis Of The Still Decoupled With Electrical Thermal Controller
Sample calculation of the thermal analysis of the constructed pyramid cover solar still is as
follows.
Radiative loss coefficient (hrw)
hrw
=
[(Tw + 273)2 + (Tg + 273)2] [Tw + Tg + 546]
Convective loss coefficient (hcw)
hcw
=
(Pw
0.884 Tw Tg
Pg ) (Tw
268.9 x 10 3
273)
1/ 3
Pw
Evaporative loss coefficient (hew)
hew
=
16.273 x 10-3hcw
Pw
Pg
Tw
Tg
3.1
External heat transfer
Since the system is well insulated at the bottom and sides the computation of external heat
transfer only radiative and convective losses are considered
The radiative heat transfer co-efficient is given by,
hra
=
(Tg
g
273) 4
(Tg
(Tsky
273) 4
Tamb )
3.2 Determination Of Thermal Efficiency ( )
The thermal efficiency of the still is also observed as well as predicted decoupled with electrical
thermal controller.
Predicted Efficiency
pre
=
Me x L
x 100
(t) x A x t
Observed Efficiency
obs
68
=
Me x L
x 100
(t) x A x t
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Vol. 04 issue 08 April 2012
3.3 PHYSICAL AND CHEMICAL ANALYSIS OF WATER BEFORE AND AFTER SOLAR
DESALINATION
Table(1) Physical examination
S.No.
Physical parameters
Test results for tap water
sample
Test results of solar
distillate sample
Clear
Clear
Colourless
Colourless
None
None
2
1
1.
Appearance
2.
Colour (Pt – Co Scale)
3.
Odour
4.
Turbidity NT Units
5.
Total dissolved solids (mg / )
118
25
6.
Electrical conductivity
(micromho/cm)
168
36
Table (2): Chemical examination
S.No.
69
Chemical parameters
Test results for tap water
sample
Test results of solar
distillate sample
7.48
7.45
1.
pH
2.
Alkalinity pH
0
0
3.
As CaCO3
42
9
4.
Total hardness as CaCO3
45
11
5.
Calcium as Ca
12
3
6.
Magnesium as Mg
4
1
7.
Sodium as Na
15
3
8.
Potassium as K
1
0
9.
Iron as Fe
0
0
10.
Manganese as Mn
0
0
11.
Free Ammonia as NH3
0
0
12.
Albuminoid Ammonia as NH3
0
0
13.
Nitrite as NO2
0
0
14.
Nitrate as NO3
2
1
15.
Chloride as Cl
22
4
16.
Fluorid as F
0.2
0
17.
Sulphate as SO4
6
2
18.
Phosphate as PO4
0
0
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Vol. 04 issue 08 April 2012
The physical and chemical analysis for the tap water and distilled water were made at the Regional
Laboratory of Tamil Nadu Water Board (TNWB), Coimbatore. The samples were tested for colour,
turbidity, total dissolved solids, electrical conductivity, pH, alkalinity chloride, fluoride, sulphate,
phosphate and the results are tabulated in the tables (3.17-3.18)
3.4 Techno – Economic Analysis
The simple techno – economic analysis (Tiwari and Yadav, 1987) of the effectiveness of solar
distillation systems considers the capital cost of the system ‗P‘ and the rate of capital recovery ‗C‘. The
first annual cost of the system A can be determined by the following formula.
A
=
Pr
(1 r)n
(1 r)n 1
Still coupled with electrical thermal controller:
Cost of the Electrical thermal controller = Rs.1000
with heating coil of 1500W
If usage of electrical back up is 1 hour/day
Then, The approximate usage of electricity, unit/month =45 units
Therefore ,
For unit /day =1.5 units
Units of electricy utilized for 3 ½ hours /day =5.25 units
for 1 unit electricity, Electricity Board charges= Rs.6.50
Therefore for 5.25 units /day = 5.25 units x 6.50
=Rs.34.125
Cost of 5.25 units/month =Rs.1023.75/month
Cost of 5.25 units/year =Rs.12455.625 /year
Total annual cost of the still without electrical =Rs.995
temperature controller per year
Total annual cost of the still when coupled with electrical =Rs.1000 +995+12455.625
temperature controller per year
= Rs.14450.625
The distillate output yield for both solar
radiation utilizing solar still and still with
electrical temperature controller is more or less
same. But the Cost of the solar still utilizing
electricity for evaporation is 15 times greater
than that of the solar still with the use of solar
radiation. This analysis shows that solar still
70
which is exposed to solar energy is highly
economical and profitable.
Graphical Analysis
The graphs are plotted between the various
observed parameters such as solar insolation,
efficiency, distillate output, temperature profiles
of various junctions etc, during selected sunny
days of experimentations.
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Vol. 04 issue 08 April 2012
Fig (4.1) shows the variation of temperature
profiles with time, still decoupled with electrical
thermal controller. The cover temperature of the
still increases as the day progresses because of
the increased evaporation of water from basin
and consequent condensation of water vapour at
the bottom surface of the top cover. Ambient
temperature variation depends on atmospheric
condition. The temperature of water and glass
cover increases in the morning hours to
maximum values around noon time before they
start to decrease late in the afternoon. This is due
to the increase of solar insolation in the morning
and the decrease in the afternoon.
Distillate output with respect to time increases in
the morning and then decreases in the afternoon
because of varying solar insolation. It is found
out in the Fig (4.2)
The variation of insolation with respect to time
shown in the Figure (4.3) it increased linearly
with time and reached the maximum value from
12 noon to 2 p.m. and then decreased. Radiation
received during this study is in the range of 0 to
1200 W/m2
The variation of the average efficiency with
respect to no. of days is drawn in Figure (4.4)
Average efficiency is varied according to solar
insolation and water collection for different
days.
Figure (4.5) shows the variation of the average
insolation with respect to no. of days. Average
insolation is varying for different days.
The water temperature and distillate output with
respect to time decoupled with electrical thermal
controller is shown in fig (4.6). Water
Temperature and the distillate yield increases in
the morning hours and starts to decrease late in
the afternoon because of decrease in solar
insolation
Figure (4.7) shows the variation of temperature
profile and distillate output with respect to time
decoupled with electrical thermal controller.
Temperature and the distillate yield increases in
71
the morning hours and starts to decrease late in
the afternoon because of decrease in solar
insolation.
Figure (4.8) shows the histogram for water
temperature with respect to distillate output for
the Pyramid still decoupled with electrical
thermal controller.
Figure (4.9), (4.10), (4.11) shows the histogram
for water temperature with respect to distillate
output for the Pyramid still coupled with
electrical thermal controller.
The histogram for water temperature with
respect to distillate output for the Pyramid still
decoupled and coupled with electrical thermal
controller is found out Table (3.15) .This shows
that both studies give approximately distillate
output for corresponding temperatures.
In general Still performance which is decoupled
with electrical thermal controller is reasonable
with a good daily output. The still coupled with
electrical thermal controller is cost effective,
even though it gives daily output similar to the
conventional still.
CONCLUSION
Solar still with pyramid cover is investigated for
its performance in this study for a number of
days. Temperature profiles of water, ambient,
cover, air, absorbent, amount of distillate output
including nocturnal output were observed. The
performance of the still with and without
electrical thermal controller is analyzed, for
augmenting the productivity. The instantaneous
and daily efficiency were calculated. Heat
transfer coefficients were computed. Numerical
calculations are carried out for comparing the
observed values of heat transference with
theoretical values. Techno economic analysis is
estimated for the system reliability. The quality
of distilled water yield is examined by a physical
and chemical properties test carried out on a
collected sample of water in the Regional
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Laboratory, Tamilnadu Water Supply and
Drainage Board, Coimbatore.
1.
2.
3.
4.
5.
6.
72
REFERENCES
Dwivedi V.K. Tiwari G.N. (2008),
Comparison of internal heat transfer
coefficients in passive solar stills by
different thermal modes: An experimental
validation , Desalination, Vol. 246, Issue. 13, Pp 304-318
Phadatare M.K. and verma S.K.(2007),
Influence of water depth on internal heat and
mass transfer in a plastic solar still,
Desalination, Vol. 217, Issue.1-3, Pp 267275
Salah Abdallah, Omar Badran and Mazen
M. Abu-Khader(2007), Performance of a
modified design of single slope solar still,
Desalination, Vol. 219, Issue 1-3, Pp 222230
Badawi W. Tleimat and Everett D. Howe
(2003), Nocturnal production of solar
distillers, Solar Energy, Vol.10, Issue. 2, Pp.
61-66
Nikolai Nijegorodov, Pushpendra K. Jain
and Stig Carlsson (2003), Thermalelectrical, high efficiency solar stills,
Renewable Energy Vol. 4, Issue 1,
Pages123-127
Boukar M and Harmim A. (2001), Effect of
climatic conditions on the performance of a
simple basin solar still: a complete study,
Desalination, Vol. 137, Issue. 1-3, Pp. 15-22
7.
8.
9.
10.
11.
12.
13.
Madhuri and Tiwari G.N.( 2001),
Performance of solar still with intermittent
flow of waste hot water in the basin,
Desalination, Vol. 52, Issue 3, Pp. 345-357
Ahmad Taleb Shawaqfeh and Mohammed
Mehdi Farid (2000). New development in
the theory of heat and mass transfer in solar
stills, Solar Energy Vol. 55, Issue 6, Pp.
527-535
Adhikari R.S. Ashvini kumar and Grag H.P.
(2000) ,Techno- economic analysis of a
multi- stage stracked tray(MSST) solar still,
Desalination, Vol. 127, Issue.1-1, Pp 19-26
Faten H. Fahmy (1998), Efficient Design of
Desalination System Using Photovoltaic and
Packed Bed Systems , Energy Sources, Part
A: Recovery, Utilization, and Environmental
Effects, Vol. 20, Issue 7 , Pp 615 – 629
Minasian A.N., Al-Karaghouli A.A., Hasan
M. and Shakir A. (1992), Utilization of solar
earth waterstills for desalination for ground
water, Energy conversion management, Vol.
49, Issue 2, Pp 107-110
Ali H.M. (1991), Mathematical model of the
solar still performance using forced
convection with condensation process
outside the still, Energy conversion
management, Vol. 1, Issue 5/6, Pp. 709-712
Lawerence S.A. and Tiwari G.N. (1991),
performance of a green house cum solar still
for the climatic condition of Port Moresby,
Energy conversion management, Vol. 1,
Issue. 2, Pp. 249-255
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Temperature profile (°C)
70
60
50
40
30
20
10
0
Time of the day
Room
Water
Absorbant
Air
Top cover
Fig. (4.1) Time Vs Temperature profile (16.12.10)
250
Distillate output (ml)
200
150
100
50
0
Time of the day
Fig. (4.2) Distillate Output Vs. Time (Date: 09.01.2011)
73
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Vol. 04 issue 08 April 2012
Fig. (4.3) Insolation vs. Time (Date: 20.01.2011)
Average efficiency
(%)
30
25
20
15
10
5
0
1
2
3
4
5
6
7
8
9
10 11 12
Number of days
Average insolation
(W / m2)
Fig.(4.4) Number of Days Vs. Average Efficiency
1000
800
600
400
200
0
1
2
3
4 5 6 7 8 9 10 11 12
Number of days
Fig. (4.5) Number of Days Vs. Average Insolation
74
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
60
50
40
30
20
10
0
0.25
0.2
0.15
0.1
0.05
0
Distillate output (kg)
Temperature profiles (°C)
Fig. (4.6) Water Temperature / Distillate Output Vs. Time (Date: 08.11.2011)
Time of the day
Distillate output
Room
Water
Air
Absorbant
Top cover
Fig. (4.7) Temperature Profile / Distillate Output Vs. Time (Date : 03.02.2011)
Fig. (4.8) Water Temperature Vs. Distillate Output (Date : 11.02.2011)
(With solar energy)
75
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Distillate output
(ml)
Fig. (4.9) Water Temperature Vs. Distillate Output
(With electrical energy)( Date: 02.03.2011)
150
100
50
0
49
50
50
50
Water Temperature (°C)
Fig. (4.10) Water Temperature Vs. Distillate Output
(With electrical energy) (Date: 03.03.2011)
Fig. (4.11) Water Temperature Vs. Distillate Output
(With electrical energy) (Date: 06.03.2011)
76
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
GEOMAGNETIC STORMS ASSOCIATED WITH IV-RADIO
BURSTS AND THEIR RELATION WITH SOLAR AND
INTERPLANETARY PARAMETERS
P.L.Verma
ijcrr
Vol 04 issue 08
Category: Research
Received on:26/02/12
Revised on:01/03/12
Accepted on:05/03/12
Department of Physics Govt. Vivekanand P.G.College Maihar Distt Satna M.P.
E-mail of Corresponding Author: verma2003@yahoomail.com
ABSTRACT
I have studied geomagnetic storms (Dst ≤-75nT), associated with Type IV radio bursts, observed during
the period 1997 to 2007 with solar and interplanetary parameters. We have observed 33 geomagnetic
storms associated with type IV radio bursts, out of which most of the geomagnetic storms (85.00%) are
intense or sever geomagnetic storms. All the geomagnetic storms are found to be associated with halo and
partial halo coronal mass ejections (CMEs).The association rates of halo and partial halo CMEs are 27
(81.82%) and 06 (18.18%) respectively .All the type IV radio bursts associated geomagnetic storms are
found to be associated with X-ray solar flares of different categories .The association rates of
geomagnetic storms with different types of flare related CMEs are found 08 (24.24)% X class flare
related CMEs,15 ( 45.46% ) M class flare related CMEs ,08 (24.24 )% C class flare related CMEs and
02(6.06)% B class flare related CMEs. Some of the type IV radio bursts associated geomagnetic storms
are found to be associated with magnetic clouds 13 (39.39 %).Majority of the geomagnetic storms are
found to be related with interplanetary shocks 31 (93.94) .We have inferred that geomagnetic storms are
closely related to interplanetary magnetic fields. We have determined positive co-relation with correlation
coefficient 0.70 between magnitude of geomagnetic storms and maximum value of IMF and 0.77 between
magnitude of geomagnetic storms and magnitude of maximum value of southward component of
interplanetary magnetic fields (IMF Bz).
____________________________________________________________________________________
Keywords –Coronal mass ejections, X-ray solar
flares, radio bursts, solar wind plasma
parameters and geomagnetic storms.
INTRODUCTION
The solar disc exhibits a variety of drastic solar
features, many of which have a direct influence
on
interplanetary
space
and
earth‘s
magnetosphere.
Coronal
mass
ejections
associated with solar flares, filaments, and type
II and type IV radio burst are the most energetic
solar features which derive solar wind
77
disturbances and these disturbances cause
geomagnetic disturbances at the earth. The
recurrent storm activity is due to coronal holes
that cause fast solar wind streams
[1].Geomagnetic storms which are characterized
by a prolonged period in which the horizontal
component of geomagnetic field is depressed in
the mid to low latitudes in the range of several
tens to several hundred nT with such periods
lasting from one-half to several days and are
classified as recurrent (periodic) and non
recurrent (sporadic). Recurrent geomagnetic
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
storms occur most frequently in the declining
phase of the solar cycle when equatoward
extensions of the polar coronal holes are
prominent [2, 3]. The non recurrent (sporadic)
geomagnetic storms are caused by interplanetary
disturbances driven by fast coronal mass
ejections (CMEs) and typically involve an
encounter with both interplanetary shock wave
and the CME that drive it. Since the CME rate
tracks the sun spot cycle [4], the non recurrent
geomagnetic storms occur most frequently near
solar maximum.Landi and Moreno et al (5) have
investigated the role of the coronal mass
ejections
in
producing
non
recurrent
geomagnetic storms in period 1969-1974. They
have concluded that coronal mass ejections
associated
with
chromospheric
flares,
accompanied by type IV radio emission, are the
most effective in perturbing the geomagnetic
field. Webb et al (6) have studied geomagnetic
storms with halo coronal mass ejections and
concluded that halo coronal mass ejections are
very much effective in producing geomagnetic
storms.Yurchyshyn [7] have analyzed data for
major geomagnetic storms and found a
relationship between hourly averaged magnitude
of the Bz component of IMF and projected
speed of CMEs launched from the central part of
the solar disk. They have concluded that CMEs
with V> 1000 Km/s are capable of furnishing.
Lepping (8) has studied geomagnetic storms
with southward directed magnetic field; they
have determined that intense geomagnetic
storms are caused by intense southward directed
magnetic field. They have found high co-relation
between Bz and Dst index. Zhag et al [9] have
studied major geomagnetic storms for the period
1996-2000 with coronal mass ejections and
concluded that 59% major geomagnetic storms
are associated with front side halo (FSH) CMEs
and 22% are associated with multiple FSH
CMEs. The events which are associated with
78
multiple FSH CMEs show complex solar wind
flows and complex geomagnetic storms which
are probably the result of halo CMEs interacting
in interplanetary space. 15% events are found to
be associated with partial halo gradual CMEs
emerging from the east limb. Michalek, and
Gopalswamy et al [10] have studied
geomagnetic storms with properties of halo
coronal mass ejections (H-CMEs) and concluded
that only fast halo CMEs (with space velocity
higher than 1000 km/s) an originating from the
western hemisphere close to the solar centre
could cause intense geomagnetic storms.
Gopalswamy et al [11] have studied
geoeffectiveness, speed, solar source, and flare
association of a set of 378 halo coronal mass
ejections (CMEs) of solar cycle 23 (1996-2005).
They have compiled the minimum Dst values
occurring within 1 - 5 days after the CME onset.
They have compared the distribution of such Dst
values for the subset of halo CMEs as disk
halos, limb halos, and back side halos CMEs.
Defining that a halo CME is geoeffective if it is
followed Dst < -50 nT, moderately geoeffective
if -50nT < Dst < -100nT, and strongly
geoeffective if Dst < -100nT, they have found
that the disk halos are followed by strong
geomagnetic storms, limb halos are followed by
moderate storms, and back side halos are not
followed by significant storms.
Experimental Data
In this investigation hourly Dst indices of
geomagnetic field have been used over the
period 2003 through 2007 to determine onset
time, maximum depression time, magnitude of
geomagnetic storms. This data has been taken
from the NSSDC omni web data system which
been created in late 1994 for enhanced access to
the near earth solar wind, magnetic field and
plasma data of omni data set, which consists of
one hour resolution near earth, solar wind
magnetic field and plasma data, energetic proton
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
fluxes and geomagnetic and solar activity
indices. The data of coronal mass ejections
(CMEs) have been taken from SOHO – large
angle spectrometric, coronagraph (SOHO /
LASCO) and extreme ultraviolet imaging
telescope (SOHO/EIT) data. To determine
disturbances in interplanetary magnetic fields,
hourly data of interplanetary magnetic field has
been used and these data has also been taken
from
omni
web
data
(http;//omniweb.gsfc.nasa.gov/form/dxi.html)).
The data of X ray solar flares radio bursts,
prominences and other solar data, solar
geophysical data report U.S. Department of
commerce, NOAA monthly issue and solar STP
data
(http://www.ngdc.noaa.gov/stp/solar/solardatase
rvices.html.) have been used. The data of
interplanetary shocks has taken from shocks
arrival derived by WIND group from WIND
observations, ACE list of transient and
disturbances.
Table-1-Association of Geomagnetic Storms with IV-Radio Bursts with Coronal Mass Ejections, X Ray Solar Flares and Magnetic Clouds for the Period of 1997-2007
Geomagnetic
Storms
S0lar Flares
Date
Onset
time in
dd(hh)
Magnit
ude in
nT
Start time
in dd(hh)
typ
es
H/
P
1
22.11.97
22(10)
-106
19(12.27)
H
2
07.11.98
07(11)
-139
05(02.02)
H
3
08.11.98
08(20)
-126
05(20.24)
4
24.05.00
08.06.00
24(00)
08(15)
-151
-89
7
17.09.00
10.11.00
17(20)
10(07)
8
27.03.01
9
Magnetic Clouds
Class
Typ
e
Start
time in
dd(hh)
19(14)
C-16
19(20.17)
IV
22(15)
3
05(03)
C-54
05(20.15)
IV
na
na
H
05(19)
M-84
05(20.15)
IV
08(23)
1
22(01.50)
06(15.54)
H
22(01)
C-63
22(01.28)
IV
na
na
H
06(15)
X-23
06(14.49)
IV
na
na
-197
-102
16(05.18)
08(04.50)
H
16(04)
M-59
16(04.33)
IV
18(02)
3
H
08(06)
C-52
08(22.51)
IV
na
na
27(21)
-86
24(20.50)
H
24(20)
M-17
24(20.20)
IV
na
na
31.03.01
31(04)
-379
28(01.27)
H
28(02)
M-17
28(11.13)
IV
na
na
10
11.04.01
11(15)
-269
09(15.54)
H
09(15)
M-79
09(15.58)
IV
12(08)
2
11
18.04.01
18(01)
-106
15(14.06)
P
15(13)
X-144
15(14.06)
IV
na
na
12
28.10.01
28(01)
-142
25(15.26)
H
25(15)
X-13
25(15.05)
IV
na
na
13
31.10.01
31(14)
-104
30(03.30)
P
30(03)
C-60
30(13.50)
IV
31(21)
3
14
05.11.01
05(19)
-297
04(16.35)
H
04(16)
X-10
04(16.12)
IV
na
na
15
17.04.02
17(11)
-149
15(03.50)
H
15(03)
M-12
15(04.32)
IV
18(04)
1
16
01(23)
02(02)
-105
-85
29(23.30)
31(02.30)
P
29(23)
C-42
29(11.00)
IV
01(12)
3
17
01.08.02
02.06.03
H
31(02)
M-93
31(02.18)
IV
na
na
18
16.06.03
16(10)
-136
14(05.30)
P
14(05)
M-15
15(23.45)
IV
na
na
19
28.10.03
28(06)
-384
27(08.30)
P
27(08)
M-27
27(08.19)
IV
na
na
20
20.11.03
20(02)
-461
18(08.05)
H
18(09)
M-45
18(08.11)
IV
20(11)
2
21
22.07.04
22(00)
-106
20(13.31)
H
20(12)
M-86
20(12.26)
IV
22(15)
3
22
07.11.04
07(20)
-376
04(09.54)
H
04(09)
C-63
04(08.49)
IV
08(03)
2
6
Start
time in
dd(hh)
Radio burts
Start time
in dd(hh)
S.
NO.
5
79
CMES
Qualit
y
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
23
07.01.05
07(12)
-94
05(15.30)
H
05(04)
B-82
04(11.03)
IV
na
na
24
16.01.05
16(20)
-117
15(06.30)
H
15(06)
M-86
15(22.33)
IV
na
na
25
21.01.05
21(19)
-103
19(08.29)
H
19(08)
X-13
19(08.12)
IV
na
na
26
15.05.05
15(05)
-293
13(17.12)
H
13(16)
M-80
13(17.03)
IV
15(06)
2
27
20.05.05
20(04)
-101
16(13.50)
P
16(13)
C-12
16(13.50)
IV
20(07)
2
28
29.05.05
29(22)
-150
26(15.06)
H
26(13)
B-75
26(21.44)
IV
na
na
29
10.07.05
17.07.05
10(11)
17(06)
-100
-77
07(17.06)
14(10.54)
H
07(16)
M-49
07(13.10)
IV
na
na
30
H
14(10)
X-12
14(10.23)
IV
na
na
31
24.08.05
24(08)
-219
22(01.31)
H
22(01)
M-26
22(01.00)
IV
na
na
32
11.09.05
11(02)
-127
09(19.48)
H
09(19)
X-62
09(19.48)
IV
na
na
33
14.12.06
14(21)
-143
13(02.54)
H
13(02)
X-34
13(02.47)
IV
14(23)
3
Table-2- Association of Geomagnetic Storms Associated with IV-Radio Bursts with Interplanetary
shocks and Interplanetary Magnetic Field for the Period of 1997-2007
Geomagnetic
Storms
S.
NO.
Onset
time in
dd(hh)
IMFBz
(nT)
IMF (nT)
Magnitude
in nT
Start
time in
dd(hh)
Start time
in dd(hh)
Magnitude
of
maximum
IMF in nT
Start time
in dd(hh)
Magnitude
of
maximum
IMFBz in
nT
1
22.11.97
22(10)
-106
22(09)
21(19)
27.1
22(20)
-12.8
2
07.11.98
07(11)
-139
07(08)
07(19)
35.4
07(22)
-19.7
3
08.11.98
08(20)
-126
08(05)
07(20)
35.4
07(20)
-11.6
4
24.05.00
08.06.00
24(00)
08(15)
-151
-89
24(17)
23(15)
08(05)
32.1
24.9
23(16)
08(13)
-24.1
-6.9
17(20)
10(07)
-197
-102
17(17)
7
17.09.00
10.11.00
10(06)
17(14)
10(05)
39.5
17.8
17(15)
10(08)
-23
-8
8
27.03.01
27(21)
-86
27(01)
27(15)
25.1
27(19)
-8.4
9
31.03.01
31(04)
-379
31(00)
30(21)
47.1
31(03)
-44.7
10
11.04.01
11(15)
-269
11(14)
11(09)
34.5
11(18)
-20.5
11
18.04.01
18(01)
-106
18(00)
17(23)
23.8
17(20)
-19.6
12
28.10.01
28(01)
-142
28(03)
27(22)
19.5
27(22)
-14.5
13
31.10.01
31(14)
-104
31(14)
31(18)
13.9
31(19)
-13
14
05.11.01
05(19)
-297
06(02)
05(12)
65.6
06(18)
-64
15
17.04.02
17(11)
-149
17(11)
17(07)
30.4
17(07)
-18.1
16
01(23)
02(02)
-105
-85
01(05)
01(01)
14.4
01(02)
-13.2
17
01.08.02
02.06.03
na
18
16.06.03
16(10)
-136
na
01(22)
15(17)
11.4
14.4
02(04)
16(09)
-8.9
-9.7
19
28.10.03
28(06)
-384
28(02)
28(01)
19.2
28(01)
-10.2
5
6
80
Date
Shocks
08(09)
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
20
20.11.03
20(02)
-461
20(07)
20(05)
55
20(11)
-50.9
21
22.07.04
22(00)
-106
22(10)
22(14)
18.9
22(19)
-15.5
22
07(20)
07(12)
-376
-94
07(02)
23
07.11.04
07.01.05
07(09)
07(12)
07(07)
47.8
20.6
07(22)
07(20)
-44.9
-18.5
24
16.01.05
16(20)
-117
17(07)
17(07)
35.3
16(08)
-5.1
25
21.01.05
21(19)
-103
21(17)
21(15)
29.5
22(18)
-2.6
26
15.05.05
15(05)
-293
15(02)
15(01)
54.2
15(05)
-38
27
20.05.05
20(04)
-101
20(04)
20(06)
15
20(02)
-9.1
28
29.05.05
29(22)
-150
19(09)
29(01)
19.2
30(05)
-16.1
29
10(11)
17(06)
-100
-77
10(03)
30
10.07.05
17.07.05
17(01)
10(02)
17(12)
25.2
14.6
10(10)
17(21)
-13.9
-8.7
31
24.08.05
24(08)
-219
24(06)
24(04)
52.2
24(05)
-38.3
32
11.09.05
11(02)
-127
11(01)
10(21)
18.2
11(00)
-6.4
33
14.12.06
14(21)
-143
14(14)
14(11)
17.9
14(22)
-14.7
DATA ANALYSIS AND RESULTS
In this study I have observed 33 geomagnetic
storms associated with type IV type radio bursts,
occurred during the period 1997 to 2007.I have
divided observed geomagnetic storms in three
categories, geomagnetic storms Dst ≤-75nT
>100 nT as moderate, Dst≤-100 nT >200nT as
intense and Dst≤-200 nT as severe .It is found
that most of most of the type IV associated
geomagnetic storms (85.00%) are intense or
severe geomagnetic storms .I have 33
geomagnetic storms in list out of which 28
geomagnetic storms have been found to be
associated with intense or severe geomagnetic
storms .The association rates of moderate,
intense and severe geomagnetic storms have
been found 15.15%,60.60% and 24.25%
respectively. From the data analysis of observed
geomagnetic storms associated with type IV
radio bursts and coronal mass ejections, all the
geomagnetic storms associated with type IV
radio bursts have been found to be associated
with halo and partial halo coronal mass ejections
and majority of them are halo coronal mass
ejections. The association rates of halo and
81
partial halo CMEs have been found 27(81.82%)
and 06(18.18%) respectively .From the further
analysis it is observed that ,CMEs which are
associated with geomagnetic storms associated
with type IV radio bursts are related with X-ray
solar flares of different categories and majority
of them are M class solar flares . The association
rates of geomagnetic storms associated with type
IV radio bursts with different types of flare
related CMEs are found 08(24.24)% X class
flare related CMEs,15( 45.46% ) M class flare
related CMEs ,08(24.24 )% C class flare related
CMEs and 02(6.06)% B class flare related
CMEs respectively .The data analysis of
observed geomagnetic storms associated with
type IV radio bursts and magnetic clouds, some
of the geomagnetic storms associated with type
IV radio bursts have been found to be associated
with excellent, good and poor quality magnetic
clouds 13 (39.39 %). Majority of the
geomagnetic storms associated with type IV
radio bursts are found to be related with
interplanetary shocks 31 (93.94) .From the data
analysis of geomagnetic storms associated with
type IV radio bursts and interplanetary magnetic
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
field ,I have found that geomagnetic storms
associated with type IV radio bursts are closely
related to
disturbances in interplanetary
magnetic fields and southward component of
interplanetary magnetic field. Further to see how
the magnitude of geomagnetic storms associated
with type IV radio bursts are correlated with the
maximum values of Jimf events, a scatter
diagram have been plotted between the
magnitude of geomagnetic storms associated
with type IV radio bursts and maximum value of
Jimf events in fig.1.From the fig it is clear that
maximum geomagnetic storms associated with
type IV radio bursts
which have large
magnitude are associated with such Jimf events
which have relatively large maximum value. I
have determined positive co-relation between
magnitude of geomagnetic storms associated
with type IV radio bursts and maximum value of
IMF with correlation coefficient 0.70 .Further to
see how the magnitude of geomagnetic storms
associated with type IV radio bursts are
correlated with maximum value of Jimfbz
events, a scatter diagram have been plotted
between the magnitude of geomagnetic storms
associated with type IV radio bursts and
maximum value of Jimfbz events in fig.2. From
the fig it is clear that maximum geomagnetic
storms associated with type IV radio bursts
which have large magnitude are associated with
such Jimfbz events which have relatively large
maximum values .Positive correlation with
correlation coefficient 0.77 have also been found
between and magnitude of geomagnetic storms
associated with type IV radio bursts and
maximum value of southward component (IMF
Bz) .
Figure-1 Shows Scatter plot between magnitudes of geomagnetic storms associated with type IV
radio bursts and magnitude of maximum value of IMF showing positive correlation with
correlation coefficient 0.70.
82
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Figure-2 Shows Scatter plot between magnitudes of geomagnetic storms associated with type IV
radio bursts and magnitude of maximum value of IMFBz showing positive correlation with
correlation coefficient 0.77.
CONCLUSION
From our study, most of the radio bursts related
geomagnetic storms have been identified as
intense or severe geomagnetic storms and
associated with coronal mass ejections (CMEs)
related to different types of X ray solar flares.
Majority of the geomagnetic storms associated
with type IV radio bursts are found to be related
with interplanetary shocks and some of them are
found to be related with magnetic clouds. Large
positive co-relation
have been determined
between magnitude of geomagnetic storms
associated with type IV radio bursts and
maximum value of IMF with correlation
coefficient 0.70 and magnitude of geomagnetic
storms associated with type IV radio bursts and
magnitude of maximum value of southward
83
component of IMF Bz with correlation
coefficient 0.77.These results shows that
geomagnetic storms associated with type IV
radio burst are mainly caused by coronal mass
ejections related with X- ray solar flares and
related with X-ray solar flares and their
interplanetary manifestations
(interplanetary
shocks and magnetic clouds).Further it is
concluded that disturbances in interplanetary
magnetic fields are closely related to
geomagnetic storms associated with type IV
radio bursts .
ACKNOWLEDGEMENT
The author would like to thank Prof. P.K
Shukla, S.K.Nigam and B.P.Chandra for
valuable suggestions. The author is grateful to
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
OMNIWEB and SOHO data group whose data
have been used in this study.
1.
2.
3.
4.
5.
84
REFERENCES
Crooker N.U. and E.W.
Cliver.
Geophys. Res. 99 A12 23, 383, 1994.
J.
Webb D.F. Revs. Geophys, (Supply) 33,
577, 1995
Y. Kamide J. Geophys. Res. V. 103, NO.
A8PP - 17705 - 17, 728, 1998
Webb, D.F. and R.A. Howard. J. Geophys
Res, 99, 4201, 1994.
R. Landi and G. moreno J. Geophys. Res
Vol. 103 No. A20, 553 – 20 –559 – 1998.
6.
Webb, D.F. Cliver, E.W. Crooker N.U. J.
Geophys Res. 105, 7491, 2000.
7. V. Yurchyshyn, Astrophys. J., 614, 1054,
2004.
8. Wu. C.C. and R.P. Lepping. J. Atm. Sol Terr. Phys. 67, 283, 2005.
9. G. Zhang K.P. Dere et al Astrophysical
Journal, 582, 520 – 533, 2003
10. Michalek, G. Gopalswamy, N. Lara, A :
Yashiro, S Space. Weather, Volume 4,
Issue 10th, citel ID S10003, 2006.
11. N. Gopalswamy, S.Yashiro, S. Akiyama, J.
Geophys. Res Vol. 112, A06112, dio :
1029/2006 JA 012149, 2007.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
PHYTOCHEMICALS
AROMETICUS BENTH
ANALYSIS
OF
COLECUS
Iffat Khan1, Kirti Jain2
ijcrr
Vol 04 issue 08
Category: Research
Received on:01/02/12
Revised on:23/02/12
Accepted on:14/03/12
1
2
Sarojini Naidu Govt. College, Shivaginagar, Bhopal, (M.P.)
Govt. science and commerce college, Benazir Jahangirabad, Bhopal, (M.P.)
E-mail of Corresponding Author: iffatkhan85@yahoo.com
ABSTRACT
Coleus aromaticus Benth. Of the family Lamiaceae is a large succulent aromatic herb used for flavoring
drinks and medicine. The leaves are considered as carminative, digestive, anthelmintic and diuretic. The
present study is dealt with the phytochemicals analysis of the leaf extracts. The leaf extracts with various
solvents like petroleum ether extract, Ethanolic extract and Aqueous extract were prepared and both
qualitative and quantitative tests for phytochemicals were done.It showed the presence of
saponins,carbohydrates, tannins,flavonoids,proteins,triterpenoids in various quantities.
____________________________________________________________________________________
Key words: Colecus, Ethanolic , saponins,
phytochemicals.
INTRODUCATION
About two and a half species of flowering plants
belonging to 10,500 genera and about 300
families are found in nature .of these a number
of genera are source of drugs. Many of the plant
products are important therapeutic agents, which
are represented by the various phytochemicals
like saponins,carbohydrates, tannins,flavonoids,
proteins, triterpenoids.
The present study is concentrated on colecus
arometicus Benth., belongs to family
Lamiaceae.Although phytochemicals are not
classified as nutrients, substances necessary for
sustaining life, they have been identified as
containing properties for aiding in disease
prevention.phytochemical are associated with
the prevention and treatment of at least four of
the leading causes of death in the united statescancer,Diabetes,cardiovascular disease, and
85
hypertensation. They are involved in many
processes including one that helps prevent cell
damage, prevent cancer cell replication, and
decrease cholesterol levels.
MATERIALS AND METHODS
Suitable explants will be taken from the
authenticated and identified plant taxonomist.
The identified and authenticated species were
collected quantity, dried and powdered for
further studies.
The phytochemicals present in the plant material
was extracted by the distillation method using
soxhlet apparatus. Different solvent, solvent
systems were used for the sepration of chemicals
according to the polarity (petroleum ether
extract, Ethanolic extract, aqueous extract) about
650 g of plant tissue were weighed and shade
dried for 10 days. The dried materials were
powdered and 50g of powder sample was
packed in a thimble and kept in soxhlet
apparatus. Each of the solvent was taken
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
separately for the extraction and the powdered
material was siphoned by 3 times. The whole
apparatus was kept over a heating mantle and
was heated continuously for 8 hours at boiling
ponit of each solvent. The extract was
concentrated to dryness and the residues were
transferred to a pre-weighed sample bottle and
were stored in desiccators for further studies.
―Phytochemicals screening of different plant
extracts found by soxhlat method.
in different media”
Different
biochemical parameters
like,
saponins,carbohydrates,
tannins,flavonoids,proteins,triterpenoids.
1. Test for flvonoids =Take 1 ml of the extract
and add few magnesium turnings, followed by
the addition of conc.HCL drop. Development of
pink colour indicates the presence of flavonoids
is present by ethanolic extract and aqueous
extract.
2. Terpenoids = Take 2mlof extract, dry and
dissolve in chloroform. Add a few drops of
acetic anhydride and conc.H2S04 and keep
undisturbed for few minutes. Formation of pink
colour indicates the presence of terpenids by
petroleum ether extract.
3. Test for tannins and phenolic compound =
To 1ml of the extract, add 2 drops of 5% Fecl3.
Presence of dirty green precipitate indicates the
presence of tannins and phenolic compound by
ethanolic extract and aqueous extract.
4. Test for saponins =5ml of the extract was
shaken with 5ml of distilled water and was
heated to the boiling point. Froathing indicates
the presence of saponins by ethanolic extract and
aqueous extract.
5. Total carbohydrate=weighed amount of
fresh tissue was homogenized with distilled
water. The homogenate was filtered using a two
layered cheese cloth. The filtrate was then
centrifuged at 10,000g for 15 min. The
supernatant was collected and the volume was
made up to 25 ml using distilled water .an
aliquot of sample was pippetted out and 4ml
anthrone reagent added. It was then kept in a
boiling water bath for 10 min. The tubes were
cooled and the absorbance was measured at
530nm. The amount of total carbohydrate
present was determined using the standard graph
of glucose.
Table:- Phyto chemicals analysis test of leaf extracts of colecus arometicus Benth in Petroleum
ether extract ethanolic extract aqueous extract.
Phytochemical constituents
Petroleum ether extract
Alkaloids
Triterpenoids
+
tenins & phenolic compound
Saponins
Total carbohydrates
+ = Present
- = Absent
RESULTS
Phytochemical screening of plant extracts. The phytochemical investigation of colecus
86
Ethanolic extract
Aqueous extract.
+
+
+
+
+
+
+
aromaticus Benth .leaf extract showed the
presence of only saponins,carbohydrates,
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
tannins,flavonoids,triterpenoids in the soxhlet
method .
Study showed that the extract of plant contain
most phytochemicals.
Such as
‗‘ saponins,carbohydrates,
tannins,flavonoids,proteins,triterpenoids‘‘
3.
DISCUSSION
Phytochemical compound of the plant were
analyzed. In the analysis of phytochemical
compound of colecus arometicus Benth extract
showed
the
presence
of
‗‘saponins,carbohydrates,
tannins,flavonoids,proteins,triterpenoids‘‘
arometicus used to treat asthma for a long time
with any undesirable side effect . colecus
arometicus is responsible for its pharmaceutical
value.
4.
CONCLUSION
Phytochemical
analysis showed that the
presence of terpenoids and phenolic compounds
were responsible for the effects. Phytochemicals
are also found in colecus arometicus, hence it
may be the reason for the activity.
In short, based on the present study it was
abserved that the plant extracts, colecus
arometicus in various phytochemicals like
‘saponins,carbohydrates,
tannins,flavonoids,proteins,triterpenoids‘‘
In view of the above, present study was
undertaken keeping mainly following objects:Study of phytochemicals of colecus
arometicus Benth.
Secondary metabolite analysis of
colecus arometicus Benth.
7.
REFERENCES
1. Abelson (1990) Editorial- Science 247 : 513.
2. Ajaiyeoba, E.O., Onocha, P.A. and
olarenwaju,
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Am,bujakshi, H.R., Thakkar Heena and
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Gmelina arborea Roxb leaves extract.
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Asha, M.K., prashant, D., Murali,B.,
Padmaja, R. and Amit, A. (2001)
Anthelmintic
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of Ocimum sanctum and eugenol,
Fitoterapia, 72: 669-670
Bradford, M.M. (1976) Annual Biochem.P.
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Chatterjee, K.D. (1967) Parasitology,
Protozoology and Helmintholgy, 6th Edn.,
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Calcutta. Dipak, N. Raut; Subodh C. Pal and
Subhash
Chandrashekhar
C.H.,
Latha,
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Vagdevi<H.M. and Vaiudya, V.P. (2008)
Anthelmintic activity of the crude extracts of
Ficus racemosa, 2:100-103.
Dipak, N.Raut; Subodh C.Pal and Subhash
C. Mandal (2009) Anthelmintic potential of
Dendrophthoe falcate Etting. (L.F.) Leaf.
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Farnsworth, N.R. and Morris, R.W (1976)
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Gamble, J.S. (1918) Flora of the Presidency
of Madras, Vol.II, pp. 1123-1124
Harbone, J.B. (1973) Phytochemical
methods, Chapman and Hall, London, pp.7.
Kingdom, A.D. and Balandrin, M.F. (1993)
Huan medicinal agents from plants, Amer
Chem. Symp. ser 534: 1-348.
Mandal (2009) Anthelmintic potential of
Dendrophthoe falcate Etting. (L.F.) Leaf.
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13. Manjamalai, A., Sardar Sathyajith Singh, R.,
Guruvayoorappan, C. and Berlin Grace,
V.M. (2010) Analysis of phytocontituents
and Anti microbial activity of some mediinal
plants in Tamilnadu, India. Global J.
Biotech & Biochem., 5(2) : 120 – 128
14. Mayr, V.D., Trutter, C., Santos- Dudga, H.,
Baner and Senchitt, W. (1995) Development
changes in the phenol concentrations of
golden delicious apple fruits and leaves. J.
Phyto.Chem., 38(5) : 1151 – 1155.
15. Minija Janardhanan and John E. Thoppil
(2004). Herb and spice essential oils
(therapeutic, flavour and aromatic chemicals
of Apiaceae). Discovery publishing house,
New Delhi.
16. Patra, A., Jha, S., Murthy, N.P. and
Vaibhav, A.D. (2008) Anthelmintic and
antibacterial activities of Hygrophila spinosa
T. Anders. Research J. Pharm and Tech. (4)
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17. Pelezcar, M.J. (1957) Microbiological
methods by the society of American
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Improving the assessment of the economic
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parasitolgy., 84:145-168.
19. Pessoa, L.M., Moris, S.M., Bevilaqna,
L.M.L. and Luciano, J.H.S. ―Anthelmintic
activity of essenbtial oil of Ocimum
gratissimum Linn. and eugenol against.
Haemonchus
controtus‖.
Veterinary
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(1987) in. to Practical Manual in
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23. Srinivasa Rao, D., Prasada Rao, V. and
Samvisiva
Rao,
K.R.S.
(2010)
Pharmacological effects of forskolin isolated
from Coleus aromaticus on the lung damage
rats. An international Journal of Advances in
Pharmaceutical Sciences. Vol (1) Sep-Oct.
24. Soodabeh Sacidina, Ahmad Reza Gohari,
Fumiyuki kiuchi and Gisho Honda (2005) in
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Channabasappa
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Mahajanshetty (2006) in vitro Anthelmintic
proerty of various seed oils. Iranian Jouranal
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27. Updegroff, D.M. (1969) Anal iochem:
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29. Zafar iqbal; Muhammad Lateef; Abdul
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Pharmaceutical Biology, Vol.44, pp. 563567
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
TOXICOLOGICAL EVALUATION OF AQUEOUS LEAF
EXTRACT OF SENNA ALATA IN PREGNANT WISTAR
RATS
Yakubu M. T.1, Adeshina A. O.1, Ibrahim, O. O. K.2
ijcrr
Vol 04 issue 08
Category: Research
Received on:29/11/11
Revised on:23/12/11
Accepted on:25/02/12
1
Phytomedicine, Toxicology and Reproductive Biochemistry Research
Laboratory, Department of Biochemistry, University of Ilorin, Ilorin,
Nigeria
2
Department of Pathology, University of Ilorin, Nigeria
E-mail of Corresponding Author: tomuyak@yahoo.com
ABSTRACT
Objective: Aqueous leaf extract of Senna alata at 250, 500 and 1000 mg/kg body weight was
evaluated for toxicity in pregnant Wistar rats. Methods: Pregnant rats were grouped into four
(A, B, C and D) of five animals each such that rats in groups A, B, C and D received 0.5 ml of
distilled water, 250, 500 and 1000 mg/kg body weight of the extract respectively, from days
10-18 post-coitus. Results: The extract reduced (P<0.05) the activities of alkaline
phosphatase, aspartate transaminase, alanine transaminase and gamma glutamyl transferase in
the liver and kidney of the animals with increases in heart and serum enzymes. The levels of
haematological parameters, serum albumin, globulin, creatinine, sodium, calcium, chloride
ions, blood urea nitrogen (BUN): creatinine ratio, total cholesterol, triacylglycerol, low- and
high-density lipoprotein cholesterol were decreased by the extract while those of urea, uric
acid, serum total bilirubin, phosphate, potassium, calcium and atherogenic index increased
significantly. The myocardial fibres were normal in the heart while there was varying degree
of necrosis of the tubular epithelial cells in the kidney and hepatic degeneration in the liver.
Conclusion: The extract caused both functional and structural toxicities and therefore not
safe for consumption during pregnancy.
___________________________________________________________________________
Keywords: Senna alata, Fabaceae,
Pregnancy, Functional toxicity, Structural
toxicity, Biomarkers
INTRODUCTION
Senna alata (Linn) Roxb. (=Cassia alata
Linn) which belongs to the Fabaceae
family (subfamily Caesalpiniaceae) is
often variously called Ringworm Bush,
Candlebra Bush, Empress Candle Plant
and Ringworm Tree (English), asunwon
oyinbo (Yoruba-Western Nigeria) and
nelkhi or okpo (Igbo-Eastern Nigeria).1 It
is native to Mexico and grows in forest
areas of West Africa. S. alata is an erect,
tropical, annual herb of 0.15 m high with
89
bilateral, leathery compound leaves (50-80
cm long) that fold together in the dark.
The fruit is a straight pod of about 25 cm
long.2 The seeds are small and square in
shape while the inflorescence looks like a
yellow candle. The root, stem, stem bark
and leaves have been separately claimed
to be used to manage hepatitis, scabies,
pruritis,
jaundice,
gastroenteritis,
ringworm, ulcer, eczema, burns, wound,
skin and upper respiratory tract infection,
diarrhoea, constipation, food poisoning
and poisonous bites.3,4 The leaves have
been implicated to be used as abortifacient
and to hasten labour.1
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
S. alata have been scientifically evaluated
for a variety of pharmacological activities
such
as
antibacterial,
antifungal,
antiparasitic, laxative, antidiabetic, antiinflammatory,
analgesic
and
abortifacient.5-12 The toxicological studies
available in the open scientific literature
on S. alata thus far have used normal mice
and rats as models.13, 14 Since the extract
have been reported to possess abortifacient
activity in pregnant rats, there is also the
need to investigate the toxic implications
of the same doses of extract in pregnant
rats. Therefore, the present study was
aimed at providing information on the
effect of aqueous leaf extract of S. alata on
some functional indices of the liver, heart
and kidney of pregnant Wistar rats. We
also investigated the haematological and
lipid
profile
as
well
as
the
histoarchitectural changes in the selected
organs of the pregnant animals following
the administration of the extract on days
10-18 post-coitus.
MATERIALS AND METHODS
Materials
Plant materials
The plant sample, obtained from herb
sellers at a market (Oja tuntun) in Ilorin,
Nigeria was authenticated at the Forestry
Research Institute of Nigeria, Jericho,
Ibadan, Nigeria. A voucher specimen (FHI
10845) was deposited at the Herbarium of
the Institute.
Assay kits and other reagents
The assay kits for urea, creatinine and uric
acid were products of Quinica Clinical
Aplicada, S.A Amosta, Spain, while those
for albumin, bilirubin, globulin, aspartate
transaminase (AST), alanine transaminase
(ALT), γ-glutamyl transferase (GGT),
sodium, potassium, calcium, chloride,
phosphate, cholesterol, triacylglycerol,
low- and high- density lipoprotein
90
cholesterol were products of Randox
Laboratories, Ltd, United Kingdom. Paranitrophenyl phosphate was a product of
Sigma-Aldrich
Chemical,
United
Kingdom. All other reagents used were of
analytical grade and were prepared in
glass-distilled water.
Laboratory animals
Forty rats (Rattus norvegicus) made up of
equal number of males (184.80 ± 4.16 g)
and females (163.65 ± 3.11 g) were
obtained from the Animal Breeding Unit
of the Department of Biochemistry,
University of Ilorin, Ilorin, Nigeria. The
animals, housed in aluminium cages that
were placed in well-ventilated room
conditions (temperature: 22 ± 3ºC; 12 h
light-dark cycle; humidity: 45%-50%)
were also supplied with rat pellets (Bendel
Feeds and Flour Mill, Ewu, Edo, Nigeria)
and water ad libitum.
Preparation of aqueous leaf extract
The leaves of Senna alata were oven-dried
at 400C for 72 h to a constant weight using
Uniscope SM9053 Laboratory Oven,
(Surgifriend Medicals, Essex, England).
The dried leaves were then pulverized
with an electric blender (Crown Star
Blender CS- 242B, Trident (H.K.) Ltd,
China) from which 100 g each was
extracted in 1000 ml of cold distilled water
for 48 hours with constant shaking. This
was later filtered with Whatmann No. 1
filter paper and thereafter lyophilized
(Micromodulyo Freeze Dryer, FS400-05,
USA) to give a yield of 16.50 g which was
reconstituted in distilled water to give the
required doses of 250, 500 and 1 000
mg/kg body weight used in this study. The
doses were as used previously in our study
on the abortifacient activity of the aqueous
leaf extract of S. alata in pregnant rats.12
Animal
grouping
and
extract
administration
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Female rats were paired overnight with the
male rats in ratio 1:1 in aluminium cages
that made the animals have free access to
food and water. The day when vaginal
plug and spermatozoa (detected with the
aid of light microscope) appeared in the
vaginal smear was considered as day zero
of pregnancy. Pregnancy was also
confirmed with the aid of pregnancy strip
dipped into their urine. The pregnant
animals were thereafter completely
randomized into four groups (A, B, C and
D) of five animals each. The distilled
water and the extracts were administered
orally to the various groups of animals on
days 10 to 18 (organogenetic period) post
coitus on daily basis, using oropharyngeal
cannula as follows: Group A (control)
received 0.5 ml of distilled water while
animals in Groups B, C and D, were
treated in the same manner as the control
except that they received equal volume of
the extract containing 250, 500 and 1000
mg/kg body weight respectively. The
animals were humanely handled according
to the guidelines of National Institute of
Health on the Care and Use of Laboratory
Animals.15 This study was carried out
following approval from the Ethical
Committee on Animal Use and Care of the
Department of Biochemistry, University of
Ilorin,
Ilorin,
Nigeria
(Ref:
UIL/BCH/EC/2008/001).
Preparation of serum and tissue
supernatants
The animals were anaesthetized in a glass
jar containing cotton wool soaked in
diethyl ether. The unconscious rat was
quickly removed and the neck area cleared
of fur. The jugular veins were cut and
blood was collected into the test tubes.
The blood samples were allowed to clot at
room temperature for 10 min after which
they were centrifuged at 894 g x 10 min.
The resulting serum was collected using
91
Pasteur pipette and kept frozen overnight
before being used for the biochemical
analyses. The animals were thereafter
dissected; the liver, heart and kidney
removed and cleaned of blood by blotting
in tissue paper. The kidneys were
decapsulated after which the organs of
interest were weighed separately and
homogenized in ice-cold 0.25M sucrose
solution (1:5 w/v). The homogenates were
centrifuged at 1398 g x 15 min to obtain
the supernatant, which were then used
within 24 hours for the biochemical
analyses.
Determination of biochemical and
haematological parameters
The procedures adopted for the
biochemical parameters were as described
for alkaline phosphatase16, aspartate and
alanine transaminases17, gamma glutamyl
transferase18, total protein19, bilirubin20,
albumin21, urea22, creatinine23, calcium,
phosphate, uric acid, globulin, potassium,
sodium and chloride ions24. The blood
urea nitrogen (BUN): creatinine ratio was
computed as the ratio of serum urea to
creatinine. The lipids assayed included
total cholesterol25, low density lipoproteincholesterol26, high density lipoproteincholesterol27,
triglycerides28
and
29
atherogenic index . The organ-body
weight ratio was computed using the
expression of Yakubu et al30. The
haematological
parameters
of
haemoglobin (Hb), packed cell volume
(PCV), red blood cells (RBC), mean
corpuscular volume (MCV), mean
corpuscular haemoglobin (MCH), mean
corpuscular haemoglobin concentration
(MCHC), white blood cells (WBC),
platelets, neutrophils and lymphocytes
were analysed using Haematologic
Analyzer, Sysmex, KX-21, Japan by
adopting the principles described by Dacie
and Lewis31.
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Vol. 04 issue 08 April 2012
Histological examination
Tissue sections of the liver, heart and left
kidney of the animals were prepared using
the procedures described by Drury and
Wallington32 and Krause33. The tissues
were stained with hematoxylin/eosin
(H&E) and photomicrographs captured at
x 400 with a Canon PowerShot A560
Digital Camera, Canon USA Inc., NY,
USA.
Statistical analysis
Data were expressed as the means + SEM
of five replicates. Significant differences
were determined using a one-way analysis
of variance and Duncan Multiple Range
Test at P<0.05.
RESULTS
The extract significantly (P<0.05)
decreased the activities of ALP, GGT,
AST and ALT in the liver and kidney of
the animals (Tables 1-4). In contrast, the
activities of these enzymes were increased
(P<0.05) by all the doses of the extract in
the heart and serum of the animals (Tables
1-4).
The extract also decreased (P<0.05) the
concentrations of albumin, globulin,
creatinine, sodium, calcium and chloride
ion in the serum of the animals whereas
the total bilirubin, urea, uric acid,
potassium and phosphate increased
(P<0.05) (Tables 5 and 6). The computed
blood urea nitrogen: creatinine ratio in the
extract treated animals was lower than the
distilled water treated control rats (Table
6).
In addition, all the doses of the extract
significantly (P<0.05) reduced the serum
concentrations of total cholesterol,
triglycerides, low- and high- density
lipoprotein cholesterols whereas the
computed atherogenic index increased
significantly (P<0.05) (Table 7).
92
The extract at the doses of 250, 500 and
1000 mg/kg body weight selectively
affected the haematological parameters.
For example, all the doses of the extract
decreased (P<0.05) the levels of Hb, PCV,
RBC, MCV, MCH, MCHC whereas the
WBC,
platelets,
neutrophils
and
lymphocytes
increased
significantly
(P<0.05) (Table 8).
While the computed heart-body weight
ratio was not significantly altered
(P>0.05), the extract decreased (P<0.05)
the liver- and kidney-body weight ratios.
Furthermore, compared with their
respective controls (Plates 1a, 2a and 3a),
the extract did not produce any
histoarchitectural change in the heart of
the animals as the myocardial fibres were
normal (Plates 1b, c and d). In contrast, the
hepatic degeneration ranged from mild to
severe with the lymphocytes extending
into the lobule in the liver (Plates 2b, c and
d) whereas in the kidney, the glomerulus
were normal with necrosis of tubular
epithelial cells and inflammatory cells
within the interstitum (Plates 3b, c and d).
DISCUSSION
Aqueous leaf extract of S. alata has been
acclaimed in folk medicine of Nigeria to
be used as an abortifacient and to ‗wash
the uterus‘. This claim however has been
substantiated by adequate scientific data.12
Previous report by Yakubu et al12 focussed
only on maternal and fetal outcomes
without reporting on the safety of the
extract in other tissues of the pregnant rats.
Therefore, the present study discusses the
implications of aqueous leaf extract of S.
alata on selected markers of damage and
histology of the liver, kidney and heart as
well as lipid and haematological profile of
pregnant rats while adopting the same
dose regimen (250, 500 and 1000 mg/kg
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
body weight) used previously in the
abortifacient study.
ALP, GGT, AST and ALT are important
markers of damage to the plasma
membrane and cytosol.34, 35 The reduction
in the activities of ALP in the liver and
kidney as well as GGT of the animals
which
was
accompanied
with
corresponding increase in the serum
enzymes suggest permeability changes
arising from damage to the cell membrane
of the organs. The elevated serum GGT, a
more effective indicator of hepatobilliary
toxicity than the ALP36
further
corroborates the hepatoxicity of the
extract. Thus, it is not surprising to have
reduction in the AST and ALT of the
tissues since damage to plasma membrane
will consequentially lead to leakage of the
cytosolic content, in this instance, the AST
and ALT. These alterations suggest that
the extract is hepato- and nephrotoxic. In
contrast, the increase in the activity of
ALP, GGT, AST and ALT in the heart of
the animals could be due to enhanced
synthesis of these enzymes. The reason for
the different pattern of toxicity pattern by
the extract on these tissues is not
immediately known but may be due to the
differences in the drug-metabolizing
enzymes of the tissues. The changes in the
activities of the enzymes will have
consequential effects on the metabolic
processes that depend on the enzymes.
The extract exhibited selective systemic
toxicity on the haematological parameters
investigated as evidenced by the decrease
in red blood cells and factors relating to it
(haemoglobin, packed cell volume, mean
corpuscular haemoglobin and mean
corpuscular haemoglobin concentration)
and increase in white blood cells and those
factors relating to it (platelets, neutrophils
and platelets). The decrease in RBC and
factors relating to it may be an indication
93
that the balance between the rate of
production and destruction has been
altered. It may also be that the individual
and total population of the red blood cells
have been adversely affected. The findings
in this study are similar to that reported
earlier by Adebayo et al37 on
Bougainvillea
spectabilis
leaves.
Furthermore, the increase in WBC and
factors relating to it suggest that the
immune system has been challenged by
the extract. These are indications of
selective systemic toxicity of the plant
extract.
Albumin and globulin are useful indicators
of synthetic function of the liver whereas
bilirubin could be used to assess excretory
function of the organ. The decrease in both
the albumin and globulin by the extract in
the present study may suggest diminished
synthetic function of the liver arising
probably from hepatocellular injury38,
increased
catabolism,
abnormal
distribution and abnormal or excessive
loss. The obtained elevated levels of
bilirubin further support hepatotoxicity of
the extract arising from an effect on the
normal excretory function of the liver of
the pregnant animals.
Changes in the serum concentrations of
creatinine, urea, uric acid and electrolytes
such as Na+, Ca2+, K+, PO42- and Cl- are
indicators of renal function at the tubular
and glomerular levels. The reduced levels
of serum creatinine by the extract suggest
glomerular dysfunction39 while similar
reduction in the levels of Na+, Ca2+ and Clindicates decreased tubular reabsorption.40
Furthermore, the increased levels of serum
urea and uric acid implies that there was
reduction in the glomerular filtration rate
of the kidney while the increase in the
levels of K+, and PO42- suggest tubular
damage since the ions are reabsorbed at
the distal tubules of the kidney. All these
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
are indications of renal dysfunction arising
probably from interference with metabolic
process of the metabolite, inefficient
filtration by the kidney and obstruction of
lower urinary tract, impaired glomerular
and tubular reabsorption or excretion of
these ions. This therefore implies
impairment or interference in the normal
excretory and reabsorptive functions of the
kidney. The serum BUN: creatinine ratio
measures the amount of nitrogen in the
blood, and can also indicate dysfunction
by either the liver or kidney since urea is
produced by the liver and excreted by the
kidney. Therefore, the low values of
computed serum BUN: creatinine ratio
compared to the control suggests that the
elevated urea in the serum of the animals
is a consequence of liver dysfunction.
Changes in the levels of cholesterol, TAG,
HDL-C and LDL-C in the serum of the
animals can serve as useful indicators of
altered lipid metabolism and predisposition
of
the
animals
to
cardiovascular risk.41 The extract affected
the metabolism of lipid in the pregnant
animals probably by impairing the normal
biosynthesis of cholesterol and enhancing
lipolysis. The significant decrease in LDLC is understandable since there is a direct
relationship between cholesterol and LDLC.41 This trend was supported in the
present study where both the cholesterol
and LDL-C of the serum of pregnant rats
were decreased by the extract of S. alata
leaves. Furthermore, the reduction in the
serum content of HDL-C, the medium by
which cholesterol from peripheral tissues
is transported to the liver to reduce the
amount stored in the tissue and the
possibility of developing atherosclerotic
plague, may not be clinically beneficial as
it may predispose the animals to
cardiovascular risk. This is corroborated
by the elevated computed atherogenic
94
index, an index of atherosclerosis and its
associated heart diseases.42
Organ-body ratio can be used to indicate
swelling, atrophy and hypertrophy.43 The
decrease in the liver- and kidney-body
weight ratios may be a manifestation of
the
moderate
to
severe
hepatic
degeneration and extensive degeneration
of tubular epithelial cells revealed by
histological examination in the present
study. In contrast, the absence of
significant changes in the heart-body
weight ratio was also corroborated by lack
of visible histoarchitectural changes since
the myocardial fibres were normal. The
nephrotoxicity and hepatotoxicity of the
aqueous leaf extract of S. alata at the
doses of 250-1000 mg/kg body weight
were corroborated by histoarchitectural
alterations in the kidney and liver of the
pregnant
animals.
Therefore,
the
toxicological impact of the aqueous leaf
extract of S. alata in the present study was
both functional and structural in the liver
and kidney whereas it was only functional
in the heart of the animals. The disparity in
the histological results of the organs may
be attributed to direct involvement of the
liver and kidney in the detoxification and
eventual elimination of the extract.
In conclusion, aqueous leaf extract of S.
alata pose toxicological risk to the organs
of pregnant rats investigated in the present
study. Therefore, the extract may cause
structural and functional dysfunctions in
the liver and kidney while the
toxicological impact is restricted to
functional dysfunction in the heart of the
pregnant animals. The extract could
predispose the pregnant animals to
systemic toxicity and cardiovascular risk.
ACKNOWLEDGEMENT
Authors acknowledge the immense help
received from the scholars whose articles
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
were cited and included in the references
of this manuscript. The authors are also
grateful to authors/editors/publishers of all
those articles, journals and books from
where the literature of this article have
been reviewed and discussed.
Declaration of Conflict of Interest or of
Financial Disclosure: The authors report
no conflict of interest. The authors alone
are responsible for the content and writing
of the paper.
1.
2.
3.
4.
5.
6.
7.
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International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table 1: Alkaline phosphatase activity of selected tissues of pregnant rats administered
with aqueous leaf extract of S. alata
_____________________________________________________________________
Alakline phosphatase activity (nm/min/mg protein)
_______________________________
Doses
Liver
Kidney
Heart
Serum
Control (Distilled water)
3.85 ± 0.07a
22.18 ±1.65a
4.76 ±0.11a
0.41 ± 0.02a
250 mg/kg body weight
2.02 ± 0.06b
12.00 ±0.82b
6.76 ±0.24b
0.78 ± 0.01b
500 mg/kg body weight
1.48 ± 0.02c
10.50 ±0.12b
7.81 ±0.03b
1.02 ± 0.03c
1000 mg/kg body weight
0.92 ± 0.05d
11.52 ±0.05b
9.02 ±0.06c
1.41 ± 0.02d
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
Table 2: Gamma glutamyl transferase activity of selected tissues of pregnant rats
administered with aqueous leaf extract of S. alata
_____________________________________________________________________
Gamma glutamyl transferase activity (U/L)
_______________________________
Doses
Liver
Kidney
Heart
Serum
Control (Distilled water)
39.52 ± 3.01a
27.83 ±5.52a
12.28 ±1.48a
4.41 ± 0.02a
250 mg/kg body weight
22.05 ± 2.18b
19.46 ±0.52b
17.26 ±1.05b
6.11 ± 0.14b
500 mg/kg body weight
11.77 ± 0.28c
12.06 ±0.16c
25.63 ±1.44c
8.76 ± 0.86c
1000 mg/kg body weight
10.59 ± 0.32c
8.14 ±1.01d
38.41 ±3.01d
8.62 ± 0.73c
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
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Table 3: Alanine transaminase activity of selected tissues of pregnant rats administered
with aqueous leaf extract of S. alata
_____________________________________________________________________
Alanine transaminase activity (U/L)
_______________________________
Doses
Liver
Kidney
Heart
Serum
Control (Distilled water)
350.00 ± 13.32a
128.46 ±5.61a
132.28 ±6.16a
32.14 ± 3.11a
250 mg/kg body weight
220.00 ± 7.96b
88.66 ±6.21b
151.50 ±1.64b
57.21 ± 4.11b
500 mg/kg body weight
139.41 ± 7.48c
62.14 ±5.72c
190.03 ±6.71c
68.00 ± 3.01c
1000 mg/kg body weight
140.19 ± 5.09c
45.82 ±4.72d
191.70 ±6.00c
88.51 ± 4.27d
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
Table 4: Aspartate transaminase activity of selected tissues of pregnant rats
administered with aqueous leaf extract of S. alata
_____________________________________________________________________
Aspartate transaminase activity (U/L)
_______________________________
Doses
Liver
Kidney
Heart
Serum
Control (Distilled water)
1010.02 ± 15.73a
880.09 ±12.44a
32.22 ±4.10a
17.28 ± 2.00a
250 mg/kg body weight
750.00 ± 10.95b
510.32 ±21.91b
49.10 ±3.88b
27.21 ± 2.11b
500 mg/kg body weight
7493.33 ± 10.41c
460.22 ±11.11c
62.86 ±6.58c
38.10 ± 3.58c
1000 mg/kg body weight
465.28 ± 8.43d
307.86 ±8.09d
64.01 ±8.62c
40.00 ± 2.08c
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
99
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Table 5: Serum liver function indices of pregnant rats administered with aqueous leaf
extract of S. alata
_____________________________________________________________________
Extract (mg/kg body weight)
_______________________________
Indices
Control
250
500
1000
Albumin (g/L)
39.00 ± 1.09a
22.00 ±0.89b
21.33 ±1.37b
14.08 ± 0.89c
Globulin (g/L)
17.50 ± 0.55a
14.00 ±1.03b
13.50 ±0.25b
9.18 ± 0.89c
Total bilirubin (g/L)
15.00 ± 0.45a
20.22 ±1.25b
22.05 ±0.67c
24.49 ± 0.59d
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
Table 6: Serum kidney function indices of pregnant rats administered with aqueous leaf
extract of S. alata
_____________________________________________________________________
Extract (mg/kg body weight)
_______________________________
Indices
Control
250
500
1000
Urea (mmol/L)
0.70 ± 0.01a
1.20 ±0.22b
1.15 ±0.05c
1.05 ± 0.05d
Creatinine (mmol/L)
19.00 ± 0.91a
14.00 ±0.19b
12.50 ±0.55c
11.50 ± 0.05c
Blood
urea
nitrogen
(BUN):creatinine ratio
1: 27
1:12
1:11
1:11
Uric acid (mmol/L)
0.03x 10-1 ± 0.79 x
10-3a
0.05 x 10-1 ± 0.89 x
10-3b
0.06 x 10-1 ± 0.89 x
10-3b
0.05 x 10-1 ± 0.52 x
10-3b
Sodium ion (mmol/L)
77.50 ± 2.73a
68.50 ±2.73b
69.00 ± 2.68b
67.00 ± 1.79b
Potassium ion (mmol/L)
1.45 ± 0.01a
1.57 ±0.04b
1.70 ±0.09c
1.73 ± 0.14c
Calcium ion (mmol/L)
1.28 ± 0.24a
1.05 ±0.02b
1.00 ±0.06b
0.95 ± 0.01b
Chloride ion (mmol/L)
0.51 ± 0.01a
0.50 ±0.00a
0.44 ±0.01b
0.26 ± 0.02c
Phosphate ion (mmol/L)
0.51 ± 0.01a
1.50 ±0.02b
0.89 ±0.01c
0.98 ± 0.01c
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
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Table 7: Serum lipid profile of pregnant rats administered with aqueous leaf extract of
S. alata
_____________________________________________________________________
Extract (mg/kg body weight)
_______________________________
Indices
Control
250
500
1000
Total cholesterol (mmol/L)
2.13 ± 0.14a
1.20 ±0.11b
1.09 ±0.09c
0.57 ± 0.04d
Triglyceride (mmol/L)
0.59 ± 0.91a
0.38 ±0.05b
0.34 ±0.06b
0.30 ± 0.02c
Low-density
lipoprotein
cholesterol (mmol/L)
High-density
lipoprotein
cholesterol (mmol/L)
1.02 ± 0.08a
0.81 ± 0.04b
0.62 ± 0.07c
0.51 ± 0.06d
2.83 ± 0.23a
1.30 ± 0.09b
1.10 ± 0.13c
0.77 ± 0.19d
Atherogenic index
(LDLC/HDLC
0.36 ± 0.13a
0.62 ±0.04b
0.56 ±0.14c
0.66 ± 0.02c
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different
(P<0.05).
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Table 8: Haematological parameters of pregnant rats administered with aqueous leaf extract of S. alata
Treatment
Distilled
water
Doses
(mg/kg
body
weight
Control
250
Extract
500
1000
Hb (g/dl)
PCV (%)
18.65
2.05a
15.50
2.81b
12.90
1.13c
11.70
1.28d
53.33
2.02a
48.33
2.31b
41.67
2.89c
31.00
2.65d
±
±
±
±
RBC
(x 1012/l)
±
4.25 ± 0.63a
±
3.37 ± 0.75b
±
3.28 ± 0.37b
±
3.27 ± 0.55b
MCV (fl)
132.33
7.57a
112.33
4.97b
113.67
6.11b
103.00
4.27c
±
±
±
±
MCH
(pg)
37.33
3.50a
23.00
0.53b
20.33
2.52b
16.67
2.29c
MCHC
(g/dl)
±
±
±
±
32.67
2.08a
24.00
0.20b
22.67
0.58b
17.67
0.13c
WBC
(x 109/l)
PLAT
(x 109/l)
±
7.77 ± 0.50a
±
9.53 ± 0.41b
±
10.53
0.15b
13.40
0.84c
±
±
±
841.00
11.01a
1013.33
17.03b
980.60
18.85b
1210.00
16.37c
NEUT
(%)
±
±
±
±
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different (P<0.05).
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10.00
0.71a
13.10
0.20b
13.67
0.02b
16.33
1.06a
LYMP
(%)
±
±
±
±
90.02
8.72a
106.00
2.80b
127.08
7.02c
122.67
6.06c
±
±
±
±
Table 9: Organ body weight ratio of pregnant rats administered with aqueous leaf extract of S.
alata
_____________________________________________________________________
Organ body weight ratio (%)
_______________________________
Doses
Liver
Kidney
Heart
Control (Distilled water)
4.69 ± 0.24a
0.63 ± 0.09a
0.36 ± 0.03a
250 mg/kg body weight
3.93 ± 0.32b
0.54 ± 0.05b
0.34 ± 0.04a
500 mg/kg body weight
3.66 ± 0.63c
0.55 ± 0.03b
0.35 ± 0.03a
1000 mg/kg body weight
3.69 ± 0.31c
0.54 ± 0.04b
0.35 ± 0.02c
Values are means ± SD of five determinations
Test values carrying superscripts different from the control are significantly different (P<0.05).
Plate 1a: Photomicrograph of the heart of pregnant rat orally administered with distilled water on
days 10-18 post coitus. The arrow shows normal myocardial fibres (x400) (H & E).
103
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Plate 1b: Photomicrograph of the heart of pregnant rat orally administered with 250 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The arrow shows normal
myocardial fibres (x400) (H & E).
Plate 1c: Photomicrograph of the heart of pregnant rat orally administered with 500 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The arrow shows normal
myocardial fibres (x400) (H & E).
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Plate 1d: Photomicrograph of the heart of pregnant rat orally administered with 1000 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The arrow shows normal
myocardial fibres (x400) (H & E).
PT
PTPT
TPT
H
Plate 2a: Photomicrograph of the liver of pregnant rat orally administered with distilled water on
days 10-18 post coitus. The arrows shows portal tract (PT) containing few lymphocytes and normal
hepatocytes (H) with no degenerative changes (x400) (H & E).
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Plate 2b: Photomicrograph of the liver of pregnant rat orally administered with 250 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. There was mild to moderate
hepatic degeneration (x400) (H & E).
Hepatic
degeneration
Plate 2c: Photomicrograph of the liver of pregnant rat orally administered with 500 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The circled spot shows portal
tract with lymphocytes extending into the lobule. The arrow shows severe degeneration of the
hepatocytes (x400) (H & E).
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Hepatic
degeneration
Plate 2d: Photomicrograph of the liver of pregnant rat orally administered with 1000 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The circled spot shows portal
tract with lymphocytes extending into the lobule. The arrow indicates severe degeneration of the
hepatocytes (x400) (H & E).
GM
Plate 3a: Photomicrograph of the kidney of pregnant rat orally administered with distilled water on
days 10-18 post coitus. The circled spot shows normal glomeruli (GM) and tubules (x400) (H & E).
107
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GM
DTEC
Plate 3b: Photomicrograph of the kidney of pregnant rat orally administered with 250 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The circled spot shows normal
glomeruli (GM) while the arrow indicates degenerated tubular epithelial cells (DTEC) (x400) (H &
E).
GM
NTEC
IFI
Plate 3c: Photomicrograph of the kidney of pregnant rat orally administered with 500 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The circled spot shows normal
glomeruli (GM) with varying degree of necrosis of tubular epithelial cells (NTEC) and
inflammatory cells within the interstitum (IFI) (x400) (H & E).
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GM
DTEC
Plate 3d: Photomicrograph of the kidney of pregnant rat orally administered with 500 mg/kg body
weight of aqueous leaf extract of S. alata on days 10-18 post coitus. The circled spot shows normal
glomeruli (GM) and extensive degeneration of tubular epithelial cells (DTEC) (x400) (H & E).
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A COMPARATIVE STUDY TO ASSESS THE CHANGES IN THE
CONDUCTION OF MEDIAN NERVE AT WRIST JOINT
IN APPARENTLY ASYMPTOMATIC COMPUTER USERS WITH
THAT IN GENERAL POPULATION
Dhaval Desai1, 2, Chintan Shah2, Harshit Soni2, Hasmukh Patel1, Komal Soni2
ijcrr
Vol 04 issue 08
Category: Research
Received on:22/03/12
Revised on:26/03/12
Accepted on:31/03/12
1
2
Shree Devi College of Physiotherapy, Mangalore
SPB Physiotherapy College, Surat
E-mail of Corresponding Author: dhavalphysio28@gmail.com
ABSTRACT
Background: Nerve Conduction Testing is frequently used by the physiotherapist as an investigation
procedure for Carpal Tunnel Syndrome. Carpal Tunnel Syndrome may develop in Computer users as well
as in General Population those who are using their wrist and finger frequently. Objectives: To compare
the Nerve Conduction changes amongst the two groups of people; those working on computer and general
population who are involved in cooking, masoning, sweeping etc. Methods: 60 individuals were divided
into 2 groups, group A and group B consisting of 30 individuals each. Group A: Individuals without any
symptoms of Carpal Tunnel Syndrome working for > 4 hours per day for > 1 year. Group B: Individuals
belonging to general population not using computer. Analysis was based on the distal motor latency and
sensory nerve action potential taken for the dominant hand. Results: Mean±SD for Distal Motor Latency
for GROUP A was 4.116±0.265ms and for GROUP B was 3.243±1.044ms. Mean±SD for Digit II to
Wrist latency for GROUP A was 2.845±0.252ms and for GROUP B was 2.077±0.556ms. Mean±SD for
Transcarpal to Wrist latency for GROUP A was 1.854±0.289ms and for GROUP B was 1.414±0.252ms.
‗t‘ calculated value for DML, Digit II to wrist and Transcarpal to wrist was 4.43, 6.88 and 6.27
respectively which was statistically significant as it is above the ‗t‘ tabulated value of 1.96.
Conclusion: There is a significant difference in both the Groups for all the parameters. GROUP A
individuals are having more chances for developing Carpal Tunnel Syndrome when compared to GROUP
B individuals.
____________________________________________________________________________________
Keywords: Carpal Tunnel Syndrome, Nerve
Conduction Velocity, Computer users and
General Population.
INTRODUCTION
Computer is an electronic device which is
omnipresent in our society; where more than
50% work is carried out by computers
(desktops/laptops). Today computers are widely
used with its basic unit like keyboard and mouse
in different sectors. About 2 of every 5
110
employed individuals are connected to use of
computers while on the job.1 As per a recent
World Bank's Enterprise Survey of 1,948 retail
stores in India, 19% of the stores use computers
for their business. In some states like Kerala,
computer use is as high as 40%.2
Repetitive strain injury (RSI); also known as
occupational overuse syndrome, non-specific
arm pain or work related upper limb disorder
(WRULD); is mainly associated with repeated
use of any particular movement like
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
flexion/extension. RSI includes conditions like
De-Quervain‘s disease, tennis elbow, carpal
tunnel syndrome, etc.
Carpal tunnel syndrome (CTS) is described as
the triad of nocturnal pain, sensory disturbance
in the distribution of the median nerve and
thenar atrophy.3 A study of employees in 21
computer companies in Chennai has revealed
that one in eight computer professionals runs the
risk of CTS and incidence increases with long
hours at computers.4 Innocuous activities such as
typing and clicking a mouse button could
possibly be harmful.
Motor nerve conduction velocity measurement
employing muscle action-potential was first
carried out by Piper (1909) and Munnich
(1916).5 The first usefulness of the median nerve
conduction studies in the diagnosis of CTS was
done by Simpson in 1956 where he
demonstrated slowing of nerve conduction in
CTS. NCV studies with a high degree of
sensitivity (>85%) and specificity (>95%)
constitute an important aspect of the diagnosis of
CTS.6 Nerve conduction change occurs before a
patient develops clinical symptoms of CTS that
are severe enough to seek medical attention.
Many asymptomatic employees can in fact be
found to have abnormalities in nerve conduction,
so by giving early intervention we can avoid
later complication and surgery for them.
In general population like sweepers, house
wives, masoning work and so likewise work
where they need to move their wrist and finger
in above mentioned direction are prone to
develop CTS in early or late stage of their life. A
study done on general population of middle part
of Italy, having population of 120,000 found that
in the 8-year period, 3,142 cases were identified
as having CTS. The mean annual crude
incidence was 329 cases per 100,000 personyears, and the standardized incidence was 276.7
A repetitive motion of wrist is one of the known
causes to develop CTS. Daily high-velocity and
111
high-force manual work is a risk factor for CTS
in a working population like Slaughter house
worker,8 Grocery store worker,9 Industries,10
Dentist11 etc. Hence the study was conducted to
assess the NCV changes in those people who
work using computers and amongst the general
population with its main objectives to evaluate
and compare the median nerve NCV changes at
wrist in computer workers who work for > 4
hours per day with those in general population.
METHODOLOGY
Study Design: Cross Sectional Comparative
Study
Study Setting: NCV Laboratory of Shree Devi
College of Physiotherapy, Mangalore
Sample size: 60 individuals
Sampling method:
The study included a sample of 60 individuals.
Out of that 30 individuals were involved in
computer work for > 4 hours per day from past 1
year and remaining 30 individuals were from
general population involved in work like
cooking, sweeping and masoning were selected
by using Convenient Sampling.
Inclusion criteria:
1) Age: between 20 years to 50 years.
2) People using laptop, desktop or both.
3) Individual working for > 4 hours/day for
a year or more.
4) Individuals engaged in cooking,
sweeping, masoning occupations.
5) Right Hand Dominants
Exclusion criteria:
1) Symptomatic Persons (CTS)
2) People with any other neurological
disorder.
3) People with any orthopedic problem.
4) People who have been operated
previously for hand.
5) Pregnant women.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
6) People with inflammatory joint disease.
i.e. Rheumatoid arthritis and obese
individuals.
Source of data collection:
1) Computer Centers.
2) House wife, Sweepers and Masons as
General population from Mangalore.
Tools used:
1. EMG machine (Neuro careTM – 2000,
computerized EMG with NCV and
Evoked potentials, Ser. No. 1023.
Manufacture Bio-techTM, India. )
Fig 1: Tool Used in Study - Neurocare 2000 EMG /NCV
Outcome measures:
1) Distal Motor Latency of Median Nerve
(DML)
2)
Sensory Latency of Median Nerve,
namely II digit to wrist latency (DII to W) and
Transcarpal to wrist latency (TC to W).
Procedure:
Prior to procedure individuals who met the
inclusion criteria were assessed and evaluated
thoroughly by using the questionnaire
(Annexure 1). After signing the consent form
they were made to participate in study.
Group A: Consists of 30 individuals without
any symptoms of CTS working for > 4 hours per
112
day for > 1 year.
Group B: Consists of 30 individuals belonging
to general population not using computer but
engaged in cooking, sweeping, masoning
occupations.
Following the above procedure the individuals
belonging to all the three groups were tested for
the NCV changes of the median nerve for
dominant hand. The Procedure for measuring
NCV was conducted as per that mentioned in
Clinical Neurophysiology, 2nd edition, by UK
Misra
&
J
Kalita.
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Vol. 04 issue 08 April 2012
Fig 2: Motor Conduction Study
Fig 3: Group A Person using Computer > 4 hours / day
Fig 4: Sensory Conduction from Digit II to Wrist
Following the recording of the above
parameters, the obtained scores were tabulated
and compared among both study groups for
NCV changes to check the probability of
occurrence of CTS amongst them.
Ethical Consideration: Procedures followed
were in accordance with the ethical standards of
Helsinki Declaration of 1975, as revised in
2000.12
Statistical analysis:
All 60 participants of both groups were analyzed
for NCV changes.
113
Fig 5: Group B General Population
Unpaired t tests were used to find out
homogeneity of two groups for all the
demographic parameters and to compare the
outcome measurement data between two groups.
Each calculated t-value is compared with t-table
value to test two tailed hypothesis at 0.05 level
of significance. Data analysis software SPSS
13.0 version has been used for the data analysis
of the present study.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
RESULTS
Table 1: shows descriptive statistics of age distribution among both groups.
Descriptive Statistics
Maximum
Mean
Std. Deviation
Std. Error of
Mean
21.00
35.00
24.9333
3.10654
.56717
21.00
35.00
26.6333
4.39030
.80156
Age (years)
N
Minimum
Group A
30
Group B
30
Graph 1: presents comparison of demographic characteristics among both the Groups.
Table 2: shows gender distribution among both the groups.
Group
Female
Gender
Male
Total
114
Group A
13
43.4%
17
56.6%
30
100%
Total
Group B
15
50.0%
15
50.0%
30
100%
28
46.6%
32
53.4%
60
100%
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table 3: Unpaired „t‟ test for age and gender
Inde pe nde nt Sam ples Te st
t-test f or Equality of Means
t
-1.731
A ge
Gender
df
58
52.219
58
58.000
1.025
Sig. (2-tailed)
.089
Mean
Dif f erence
-1.70000
Std. Error
Dif f erence
.98192
.310
.13333
.13013
Table 3 presents Unpaired ‗t‘ test which was calculated at 0.05 level of significance to compare both the
groups in terms of age and gender distribution and also to find out the homogeneity of both the groups for
comparison of outcome measures. ‗t‘ calculated value of age and gender distribution across both groups
was -1.731 and 1.025 respectively which was not significant, hence both the groups were comparable.
Table 4: Outcome Measures for Both Groups
Group Statis tics
DML
D II to W
TC to W
Group
A
B
A
B
A
B
N
30
30
30
30
30
30
Mean
4.1167
3.2437
2.8450
2.0773
1.8540
1.4140
Std. Deviation
.26583
1.04414
.25268
.55608
.28964
.25220
Std. Error
Mean
.04853
.19063
.04613
.10153
.05288
.04604
Table 4: shows outcome measures for both the groups. Mean±SD for Distal Motor Latency for GROUP
A was 4.116±0.265ms and for GROUP B was 3.243±1.044ms. Mean±SD for Digit II to Wrist latency for
GROUP A was 2.845±0.252ms and for GROUP B was 2.077±0.556ms. Mean±SD for Transcarpal to
Wrist latency for GROUP A was 1.854±0.289ms and for GROUP B was 1.414±0.252ms.
115
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Graph 2: presents comparison of outcome measures among both the Groups.
Table 5:„t‟ calculated value for outcome measures of both the groups
Inde pe nde nt Samples Te s t
Levene's Test for
Equality of Variances
F
9.303
Sig.
.003
DII to W
16.126
.000
TC to W
.180
.673
DML
t-test for Equality of Means
t
4.438
4.438
6.884
6.884
6.275
6.275
df
58
32.744
58
40.486
58
56.923
Sig. (2-tailed)
.000
.000
.000
.000
.000
.000
Mean
Difference
.87300
.87300
.76767
.76767
.44000
.44000
Std. Error
Difference
.19671
.19671
.11151
.11151
.07012
.07012
95% Confidence
Interval of the
Difference
Low er
Upper
.47923
1.26677
.47266
1.27334
.54445
.99089
.54237
.99296
.29964
.58036
.29959
.58041
As evident from table 5, ‗t‘ calculated value for DML, Digit II to wrist and Transcarpal to wrist was 4.43,
6.88 and 6.27 respectively which was statistically significant as it is above the ‗t‘ tabulated value of 1.96.
DISCUSSION
The study was conducted to examine the
changes of NCV in computer workers and
amongst the general population. The study was
done on 30 individuals with a mean age of 24.93
± 3.10 who used computer on daily basis > 4
hours / day for 1 year or more and remaining 30
individuals with the mean age of 26.63 ± 4.39
116
who were involved in cooking, sweeping,
masoning, etc.
Distal Motor Latency and Sensory Latency
(SNAP) from Digit II to Wrist and Transcarpal
region to wrist in dominant upper extremity
were evaluated. After retrieving the values, data
was statistically compared using Unpaired ‗t‘
test for comparison of both the groups.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Result showed that there is significant difference
in both the GROUPS when compared in terms
of DML, Digit II to Wrist Latency and
Transcarpal to Wrist Latency. In all the three
outcome parameters, Group A had significantly
higher latencies as compared to group B. So we
can conclude that there are more chances to
develop CTS in GROUP A (Computer users
those who use it for > 4 hours / day from past 1
year) compared to GROUP B. This can be
attributed to the repetitive action of fingers and
abnormal wrist postures seen during typing and
mouse operating leading to abnormal stress on
the underlying tissues especially the nerves.
One of the hallmarks of compressive
neuropathies such as CTS is demyelination,
producing the reduction in normal conduction of
neural impulses, which appears to result
primarily from the mechanical disruption of
internodal segments, extensive demyelination
and persistent compression eventually resulting
in direct axonal damage and wallerian
degeneration distal to site of injury.
Limitations of the study
 The study was done on a small sample
size.
 Only Electro diagnostic tool was used to
find out CTS.
 Only Median Nerve value was taken in
to consideration.
Scope of further studies
 Comparison of Sensory and Motor nerve
conduction velocity of median and
ulnar nerves can give better result.
 Probabilities of occurrence of CTS
based on number of hours spend/day on
computer usage can be compared in
future.
 Comparison of dominant to nondominant hand can be done to check
the probability of occurrence of CTS
among them.
117
 On basis of the acquired results,
ergonomic advice can be given to the
involved group individuals.
CONCLUSION
Study was done on Computer users and General
population involved in cooking, sweeping etc.
for assessing the NCV abnormalities and thereby
to identify presence of CTS by using parameters
like DML (Distal Motor Latency), Latency
from IInd Digit to Wrist and Transcarpal region
to Wrist. The result showed that there is a
significant difference in both the Groups for all
the parameters suggesting that among them,
GROUP A individuals i.e. computer operators
are having more chances for developing CTS
when compared to GROUP B individuals, i.e.
general population.
ACKNOWLEDGEMENTS
First and foremost we would like to thank God
to bless us enough courage and ability to pursue
this work.
We are greatly thankful to all the scholars whose
articles are cited and included in references of
this manuscript. We are also grateful to authors/
editors/ publishers of all those articles, journals
and books from where the literature for this
article has been reviewed and discussed. We are
thankful to all our subjects who participated with
full cooperation and showed voluntary interest,
without them this study would not have been
possible. Finally we are thankful to all those
who directly or indirectly contributed to this
study.
BIBLIOGRAPHY
1. United States department of labour,
Computer use at work in 2003, August 03,
2005, available at www.bls.gov.
2. Mohammad Amin. World Bank. Are Labour
Regulations Driving Computer Usage in
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
3.
4.
5.
6.
7.
India‘s Retail Stores? World Bank Policy
Research Working Paper 4274, July 2007.
S. Brent Brotzman, Kevin E.Wilk; Clinical
orthopaedic rehabilitation, 2nd edition.
Mosby. pp 34-39
Ali KM, Sathiyasekaran BW. Computer
professionals and Carpal Tunnel Syndrome
(CTS). Int J Occup Saf Ergon 2006; 12:
319-325.
UK
Misra,
J
Kalita,
clinical
neurophysiology, 2nd edition, 1st chapter,
page: 1-8.
C. K. Jablecki, M. T. Andary, M. K. Floeter
and R. G. Miller. Practice parameter:
Electrodiagnostic studies in carpal tunnel
syndrome: Report of the American
Association of Electrodiagnostic Medicine.
Neurology 2002; 58: 1589-1592.
Mauro Mondelli, Fabio Giannini, and
Mariano Giacchi. Carpal tunnel syndrome
incidence in a general population.
NEUROLOGY 2002; 58: 289–294.
118
8. Frost P, Andersen JH, Nielsen VK. (1998)
Occurrence of carpal tunnel syndrome
among slaughterhouse workers. Scand J
Work Environ Health; 24: 285-292.
9. Osorio AM, Ames RG, Jones J, et al. (1994)
Carpal tunnel syndrome among grocery
store workers. Am J Ind Med; 25: 229–245
10. Bingham RC, Rosecrance JC, Cook TM.
Prevalence of abnormal median nerve
conduction in applicants for industrial jobs.
Am J Ind Med 1996 Sep; 30(3): 355-61.
11. Werner RA, Hamann C, Franzblau A, et al.
(2002) Prevalence of carpal tunnel
syndrome and upper extremity tendinitis
among dental hygienists. J Dent Hyg; 76:
126–132
12. WMA Declaration of Helsinki - Ethical
Principles for Medical Research Involving
Human Subjects. 59th WMA General
Assembly Seoul, Korea, Oct 2008.
http://www.wma.net/en/30publications/10po
licies/b3/
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
DESIGN OF A COMPACT CPW FED HEXAGON SHAPED
SLOT ANTENNA FOR WI-MAX APPLICATION
ijcrr
Vol 04 issue 08
Category: Research
Received on:20/03/12
Revised on:25/03/12
Accepted on:30/03/12
H. M. Ramesh, K. Balaji, D.Ujwala, B.Harish, Ch. Vijay Sekhar Babu,
K.Naga Mallik
Department of ECE, K L University
E-mail of Corresponding Author: ujwala.dogiparthi@gmail.com
ABSTRACT
A compact Coplanar Waveguide (CPW) fed Hexagon shaped slot antenna is proposed for Wi-Max
application. The proposed antenna has a size of 20mm x 16.5mm x1.6mm and it is designed on Rogers
RT/Duroid substrate with a dielectric constant of 2.2. The proposed antenna resonates at 7.5 GHz and has
a bandwidth of 1.2 GHz, Return Loss (S11) of -51.5dB, Gain 3.7 dBi, VSWR of 1 at 7.5 GHz. The near
field and far field radiation patterns are bi-directional and Omni-directional in E and H planes. The
proposed antenna is used for Wi-Max applications and the antenna has an impedance bandwidth of 86%.
The proposed antenna is simulated using Ansoft High Frequency Structure Simulator (HFSS) version 13
which is based on Finite Element Method and the antenna parameters are analyzed.
____________________________________________________________________________________
KEYWORDS: Co-Planar Waveguide (CPW),
Near-Field, Far-Field, Wi-Max, Slot antenna.
INTRODUCTION
Worldwide Interoperability for Microwave
Access (Wi-Max) [1] with IEEE 802.16 standard
is an emerging wireless technology for high data
rate transfer of approximately 75Mb/s. The
design of wideband antenna with compact size,
low profile, light weight and obtaining beneficial
results for fundamental antenna parameters like
Return Loss, VSWR, Gain and Radiation
Patterns is a challenge. A slotted patch antenna
fed by a CPW structure provides broad
bandwidth with low dispersion and less
radiation. Among planar UWB antennas, slot
antennas are more preferred because of their
higher impedance bandwidth, very good
radiation efficiency and less dispersion [2-6].
119
In this paper, a compact hexagonal shaped slot
antenna is proposed which is useful for Wi-Max
[7-10] application. The proposed antenna is easy
to integrate, low profile with less radiation loss
and less dissipation. Since CPW feed
implemented here is of ungrounded type, there is
no need of ground plate and this improves the
radiation characteristics. The proposed antenna
is excited with 50 ohms CPW feed.
ANTENNA DESIGN
The geometry of the proposed CPW fed
hexagonal slot antenna is as shown in figure 1.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
The maximum Gain at resonant frequency of 7.5
GHz is 3.73 dBi and the 3D gain plot is shown
in figure 4. The Impedance Bandwidth is 86%.
Figure 1: Proposed Antenna Structure
The proposed antenna is designed on Rogers
RT/duroid 5880 substrate on one layer metallic
side. The relative permittivity of the substrate is
2.2 with a dielectric loss tangent of 0.0009. The
overall size of the antenna is 20 x 16.5 x 1.6
mm. The optimum design parameters are shown
in figure 1. The feed line width is 2mm at the
bottom and increasing towards the top and the
width at the top is 3.6 mm. The gap from the
coplanar ground plane to the patch is 1.2mm.
The length and width of the hexagonal slot is 18
x 10.3 mm. The folded slot dimensions are 6 x 1
mm with a gap of 1 mm between them. The
simulation process is carried out using Ansoft
High Frequency Structure Simulator (HFSS).
The excitation given to this antenna is Lumped
port excitation. The Return Loss and Resonant
frequency will vary with the dimensions of the
patch, coplanar ground plane and substrate.
RESULTS AND DISCUSSIONS
The proposed CPW fed hexagon shaped slot
antenna resonates at a single frequency of 7.5
GHz (7 GHz to 8.2 GHz) which has a bandwidth
of 1.2GHz. The Frequency vs. Return Loss plot
is shown in figure 2 and the return loss at the
resonating frequency is -51.5 dB. The VSWR at
7.5 GHz is 1.0 and the plot is shown in figure 3.
120
Figure 2: Return Loss – S11 vs. Frequency
Figure 3: VSWR vs. Frequency
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Figure 7: Surface current distribution
Figure 4: 3D Gain
The Electric field, Magnetic field and Surface
current distributions at 7.5GHz are shown in
figures 5, 6, 7 respectively. Mesh plot is shown
in figure 8.
Figure 8: Mesh Plot
Figure 5: E-Field Distribution
The far field E-Plane and H-Plane Radiation
patterns at resonant frequency 7.5GHz are
shown in figures 9 and 10.
Figure 6: H-Field Distribution
.
Figure 9: E-Plane Radiation Pattern at Phi=0
deg and Phi=90 deg
121
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
At 7.5 GHz
S. No rE Field
Figure 10: H-Plane Radiation Pattern at
Theta=0 deg and Theta=90 deg
Table 1 represents the output parameters of the
antenna like Peak Directivity, Radiated Power,
Radiation Efficiency etc. and obtained a
Radiation Efficiency of 99.9% which shows that
there is less loss of Radiation.
Table 2 represents the radiated field data at the
resonant frequency 7.5GHz at different Theta
and Phi. From the table, Left hand Circular
Polarization (LHCP) and Right hand Circular
Polarization (RHCP) are approximately equal.
S. No
Quantity
Value at 7.5
GHz
1
Max U
0.002
2
Peak Directivity
2.376
4
Peak Realized Gain
2.3738
5
Radiated Power
0.0099908
6
Accepted Power
0.0099999
7
Incident Power
0.01
8
Radiation Efficiency
0.99909
9
Front to Back Ratio
1.0873
Table 1: Antenna output parameters
122
Value (V)
At Phi At Theta
(deg)
(deg)
1
Total
1.2
-78
176
2
X
0.5
-50
88
3
Y
1.2
-76
178
4
Z
0.6
-90
136
5
Phi
1.1906
0
180
6
Theta
1.1933
-90
176
7
LHCP
0.873
-160
164
8
RHCP
0.8803
-18
164
Table 2: Radiation Field data
CONCLUSION
A 50 Ω CPW fed hexagon shaped folded slot
antenna of compact size 20 x 16.5 x 1.6 mm is
designed for Wi-Max application with excellent
Return Loss of -51.5 dB and desirable peak gain
of 3.7 dBi and acceptable VSWR of 1.0 at the
resonant frequency 7.5 GHz (7GHz - 8.2GHz).
The bandwidth of the designed antenna is 1.2
GHz with an impedance bandwidth of 86%. The
E-Plane and H-Plane radiation patterns for
different Phi (0 and 90 deg.) and Theta (0 and 90
deg.) values are obtained as quasi Omnidirectional
with
acceptable
Radiation
characteristics
and
desirable
Radiation
Efficiency.
ACKNOWLEDGEMENT
The authors would like to acknowledge their
gratitude towards the management of
Department ECE, K. L. University for their
support during the work. Authors acknowledge
the immense help received from the scholars
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
whose articles are cited and included in
references of this manuscript. The authors are
also grateful to authors / editors / publishers of
all those articles, journals and books from where
the literature for this article has been reviewed
and discussed.
1.
2.
3.
4.
5.
REFERENCES
Rodney B. Water house, ―Kin-Lu Wong‘
―Planar
antenna
for
wireless
communications‖ Wiley – Interscience,
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Jyh-Ying Chiou, Jia-Yi Sze, and Kin-Lu
Wong, "A broad-band CPW fed strip-loaded
square slot antenna," IEEE Trans. Antennas
Propag., vo1.51, pp. 719-721,2003.
H. D. Chen, ―Broadband CPW-fed square
slot antennas with a widened tuning stub,‖
IEEE Trans. Antennas Propagation, vol. 51,
pp. 1982-1986, 2003.
Vi-Cheng Lin, and Kuan-Jung Hung,
"Compact
ultrawideband
rectangular
aperture antenna and band-notched designs,"
IEEE Trans. Antennas Propag., vo1.54,
pp.3075-3081, 2006.
Li Y.S. , Yang X.D., Liu C.Y., and Jiang T.,
"Compact CPW-fed ultrawideband antenna
123
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10.
with
band-notched
characteristic,"
Electron.Lett. vo1.46, pp.1533-1534, 2010.
Chen x., Zhang W. , Ma R., Zhang J., and
Gao J., "Ultra-wideband CPW-fed antenna
with round comer rectangular slot and
partial circular patch," IET Microw.
Antennas Propag., vol.1, pp.847-851, 2007.
Lin Dang and et al, "A compact microstrip
slot triple-band antenna for WLAN/WiMAX
applications," IEEE Antennas Wireless
Propag. Lett., vol. 9, pp. 1178 - 1181,2010.
Wen-Shan Chen and Kuang-Yuan Ku,
"Band-rejected design of the printed open
slot antenna for WLAN/WiMAX operation,"
IEEE Trans. Antennas Propag., vo1.56,
pp.l163-1169, 2008.
Yang Zhang, Xin Sun, Jian-xing Wu, Hongchun, Gang Zeng, ―Design of a CPW-Fed
Compact Slot Antenna for WIMAX/WLAN
Applications‖, IEEE 2011.
Pichet
Moeikham
and
Prayoot
Akkaraekthalin, ―A Pentagonal Slot
Antenna with Two-Circle Stack Patch for
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International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
BIOEQUIVALENCE AND HIGHLY VARIABLE DRUGS: AN
OVERVIEW
Vikram Lohar1, Harsh Patel2, Arvind Singh Rathore1, Sandeep Singhal1,
Ashish Kumar Sharma2, Parul Sharma1
1
ijcrr
Vol 04 issue 08
Category: Review
Received on:19/02/12
Revised on:08/03/12
Accepted on:23/03/12
2
Faculty of Pharmaceutical sciences, Jodhpur National University, Jodhpur
Suresh Gyan Vihar College of Pharmacy, Jaipur
E-mail of Corresponding Author: lohar.vikram@gmail.com
ABSTRACT
Bioequivalence studies are the preliminary requirement for generic products to enter in the market. The
manufacturer (generic) must be in limit with that of innovator (branded) formulation (reference listed
drug) within the limits approved by respective governing bodies. As per biopharmaceutical classification
system the drugs falls in the category I to IV on the basis of permeability and solubility data. Drugs
belonging to the category of poor solubility and poor permeability data uphold bioequivalence issues. Due
to this high variability, large sample size may be needed in BE studies to give adequate statistical power
to meet FDA BE limits, and thus designing BE studies for HVDs is challenging. Consequently
development of generic products for HVDs is a major concern for the generic drugs industry. Major
regulatory agencies also considered different approaches for evaluating BE of highly variable drugs.
From 2004 onward the FDA started looking for alternative approaches to resolve this issue, and
eventually found that replicate crossover design and scaled average BE provides a good approach for
evaluating the BE of highly variable drugs and drug products as it would effectively decrease sample size,
without increasing patient risk.
____________________________________________________________________________________
Key words: Bioequivalence, Highly Variable
Drugs, Pharmacokinetic.
INTRODUCTION
Generic drug
According to the U.S. Food and Drug
Administration (FDA), generic drugs are
identical or within an acceptable bioequivalent
range to the brand name counterpart with respect
to pharmacokinetic and pharmacodynamic
properties. By extension, therefore, generics are
considered (by the FDA) identical in dose,
strength, route of administration, safety,
efficacy, and intended use. The FDA‘s use of the
word identical is very much a legal
124
interpretation, and is not literal. In most cases,
generic products are available once the patent
protections afforded to the original developer
have expired. When generic products become
available, the market competition often leads to
substantially lower prices for both the original
brand name product and the generic forms.
Hatch Waxman Act
Using bioequivalence as the basis for approving
generic copies of drug products was established
by the ―Drug Price Competition and Patent Term
Restoration Act of 1984,‖ also known as the
Waxman-Hatch Act. Under Hatch-Waxman Act,
one of the following four certifications has to be
made while filing an ANDA: [Food and drug
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Vol. 04 issue 08 April 2012
administration, center for drug evaluation and
research (CDER)]. 1
Bioavailability (BA) and bioequivalence (BE)
studies provide important information in the
overall set of data that ensure the availability of
safe and effective medicines to patients and
practitioners. BA and BE measures are
frequently expressed in systemic exposure
measures, such as area under the plasma
concentration-time curve (AUC) and maximum
concentration (Cmax). These measures of
systemic exposure are assumed to relate in some
way to safety and efficacy outcomes that may be
expressed in biomarkers, surrogate endpoints, or
clinical benefit end points. 2
Bioequivalence (BE) is defined as the absence of
a significant difference in the rate and extent to
which the active ingredient or active moiety in
pharmaceutical equivalents or pharmaceutical
alternatives becomes available at the site of drug
action when administered at the same molar
dose under similar conditions in an appropriately
designed study 3. BE studies of systemically
absorbed drug products are generally conducted
by determining pharmacokinetic endpoints to
compare the in vivo rate and extent of drug
absorption of a test and a reference drug product
in healthy subjects. A test product is considered
bioequivalent to a reference product if the 90%
confidence intervals for the geometric mean
test/reference ratios of the area under the drug‘s
plasma concentration versus time curve (AUC)
and peak plasma concentration (Cmax) both fall
within the predefined BE limits of 80–125% .4
The width of the 90% confidence interval is
proportional to the estimated drug variability (in
particular, within-subject variability for a
crossover design) and inversely proportional to
the number of subjects participating in the study.
The BE limits of 80–125% are currently applied
to almost all drug products regardless of the size
of within-subject variability. As a result, the
number of subjects required for a study of highly
125
variable drugs or drug products can be much
greater than normally needed for a typical BE
study. For example, to demonstrate BE with
90% power, it was estimated that 136 subjects
would be required for a drug with 60% within
subject coefficient of variation even if the test
and reference products were identical. 5
Traditional Bioequivalence Method
For systemically available drug products, FDA
generally asks applicants to conduct BE studies
with pharmacokinetic endpoints using a single
dose, crossover design in healthy subjects. The
processes of study design and workflow of
BA/BE studies are presented in brief in Figure 1
and Table 1 describes various study designs
generally used for BA/BE studies.
Subjects receive a single dose of test and
reference products on separate occasions with
random assignment to the two possible
sequences of product administration. Treatments
are separated by a washout period of adequate
duration such that the drug of interest can no
longer be detected in plasma. The FDA
generally asks applicants to conduct single dose
studies rather than multiple dose studies because
single dose studies are generally more sensitive
to detecting potential differences between
products 4. For a product with multiple strengths,
the highest strength is used in the BE study,
unless precluded for reasons of safety. The
number of subjects in the study should be
sufficient to ensure adequate statistical power;
most studies enroll from 24 to 36 subjects.
The bioequivalence parameters AUC and Cmax
are statistically analyzed using the two one-sided
tests procedure to determine whether the average
values for the measures estimated after
administration of the test and reference products
are comparable.6 This approach involves the
calculation of a 90% confidence interval for the
ratio of the averages of the measures for the test
and reference products. 7 The choice of the
current 80 to 125% acceptance limits for BE has
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Vol. 04 issue 08 April 2012
been based on expert medical judgment and
FDA experience with thousands of drug
products that a difference of less than 20% in
drug exposure was not clinically significant for
most drugs.8 The 80% limit indicates that the
test product is no less than 80% of the reference,
while the 125% limit indicates that the reference
product is no less than 80% of the test product (a
4:5 reference to test ratio is a 5:4 test to
reference ratio).
ASSESSMENT OF BIOEQUIVALENCE
The assessment of BE of different drug products
is based on the fundamental assumption that two
products are equivalent when the rate and extent
of absorption of the test/generic drug does not
show a significant difference from the rate and
extent of absorption of the reference/brand drug
under similar experimental conditions as
defined. As per the different regulatory
authorities, BE studies are generally classified
as:
1. Pharmacokinetic endpoint studies.
2. Pharmacodynamic endpoint studies.
3. Clinical endpoint studies.
4. In vitro endpoint studies.
The general descending order of preference of
these
studies
includes
pharmacokinetic,
pharmacodynamic, clinical, and in vitro studies.
Pharmacokinetic endpoint studies
These studies are most widely preferred to
assess BE for drug products, where drug level
can be determined in an easily accessible
biological fluid (such as plasma, blood, urine)
and drug level is correlated with the clinical
effect. The statutory definition of BA and BE,
expressed in rate and extent of absorption of the
active moiety or ingredient to the site of action,
emphasizes the use of pharmacokinetic measures
to indicate release of the drug substance from
the drug product with absorption into the
systemic circulation.
Regulatory guidance recommends that measures
of systemic exposure be used to reflect clinically
126
important differences between test and reference
products in BA and BE studies. These measures
include
i) Total exposure (AUC0–t or AUC0–∞ for singledose studies and AUC0–τ for steady-state
studies),
ii) Peak exposure (Cmax), and
iii) Early exposure (partial AUC to peak time of
the reference product for an immediate-release
drug product). Reliance on systemic exposure
measures will reflect comparable rate and extent
of absorption, which, in turn, will achieve the
underlying goal of assuring comparable
therapeutic effects. Single dose studies to
document BE were preferred because they are
generally more sensitive in assessing in vivo
release of the drug substance from the drug
product when compared to multiple dose studies.
The following are the circumstances that
demand multiple-dose study/steady state
pharmacokinetics:
 Dose- or time-dependent pharmacokinetics.
 For modified-release products for which the
fluctuation in plasma concentration over a
dosage interval at steady state needs to be
assessed.
 If problems of sensitivity preclude
sufficiently precise plasma concentration
measurements
after
single-dose
administration.
 If the intra-individual variability in the
plasma
concentration or disposition
precludes the possibility of demonstrating
BE in a reasonably sized single-dose study
and this variability is reduced at steady state.
 When a single-dose study cannot be
conducted in healthy volunteers due to
tolerability reasons and a single-dose study
is not feasible in patients.
 If the medicine has a long terminal
elimination
half-life
and
blood
concentrations after a single dose cannot be
followed for a sufficient time.
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Vol. 04 issue 08 April 2012
 For those medicines that induce their own
metabolism or show large intra-individual
variability.
 For combination products for which the ratio
of plasma concentration of the individual
substances is important.
 If the medicine is likely to accumulate in the
body.
 For enteric coated preparations in which the
coating is innovative.
Under normal circumstances, blood should be
the biological fluid sampled to measure drug
concentrations.
Most drugs may be measured in serum or
plasma; however, in some drugs, whole blood
(e.g., tacrolimus) may be more appropriate for
analysis. If the blood concentrations are too
minute to be detected and a substantial amount
(40%) of the drug is eliminated unchanged in the
urine, the urine may serve as the biological fluid
to be sampled (e.g., alendronic acid).
Pharmacodynamic endpoint studies
Pharmacokinetic studies measure systemic
exposure but are generally inappropriate to
document local delivery BA and BE. In such
cases, BA may be measured, and BE may be
established, based on a pharmacodynamic study,
providing an appropriate pharmacodynamic
endpoint is available. Pharmacodynamic
evaluation is measurement of the effect on a
pathophysiological process, such as a function of
time, after administration of two different
products to serve as a basis for BE assessment.
Regulatory authorities request justification from
the applicant for the use of pharmacodynamic
effects/parameters for the establishment of BE
criteria. These studies generally become
necessary fewer than two conditions
1) If the drug and/or metabolite(s) in plasma or
urine cannot be analyzed quantitatively with
sufficient accuracy and sensitivity;
2) If drug concentration measurement cannot be
used as surrogate endpoints for the
127
demonstration of efficacy and safety of the
particular pharmaceutical product. The other
important specifications for pharmacodynamic
studies include:
 A dose-response relationship should be
demonstrated;
 Sufficient measurements should be taken to
provide an appropriate pharmacodynamic
response profile;
 The complete dose-effect curve should
remain below the maximum physiological
response;
 All
pharmacodynamic
measurements/methods should be validated
for
specificity,
accuracy,
and
reproducibility.
Examples
of
these
pharmacodynamic studies include locally
acting drug products and oral inhalation
drug products, such as metered dose inhalers
and dry powder inhalers, and topically
applied dermatologic drug products, such as
creams and ointments.
Bronchodilator drug products, such as albuterol
metered dose inhalers, produce relaxation of
smooth muscle of the airways. For these drug
products, a pharmacodynamic endpoint, based
either on increase in forced expiratory volume in
1 second (FEV1) or on measurement of PD20 or
PC20 (the dose or concentration, respectively, of
a challenge agent) is clinically relevant and may
be used for BA and BE studies.
Clinical endpoint studies or comparative
clinical trials
In the absence of pharmacokinetic and
pharmacodynamic approaches, adequate and
well-controlled clinical trials may be used to
establish
BA/BE.
Several
international
regulatory
authorities
provide
general
information about the conduct of clinical studies
to establish BE.
In vitro endpoint studies
More
recently,
a
Biopharmaceutics
Classification System (BCS) has categorized
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Vol. 04 issue 08 April 2012
drug substances as having either high or low
solubility or permeability and drug products as
exhibiting rapid dissolution. According to this
approach, drug substances may be classified into
four primary groups:
1) Highly soluble and highly permeable;
2) Highly permeable and poorly soluble;
3) Highly soluble and poorly permeable;
4) Poorly soluble and poorly permeable.
Using this BCS approach, a highly permeable,
highly soluble drug substance formulated into a
rapidly dissolving drug product may need only
in vitro dissolution studies to establish BE. In
addition, in vitro approaches to document BE for
nonbioproblem drugs approved before 1962
remain acceptable as per FDA regulations.
Dissolution tests can also be used to reduce the
number of in vivo studies in other
circumstances, and to
i) Assesses batch-to-batch quality and support
batch release;
ii) Provide process control and quality
assurance; and
iii) Assess the need for further BE studies
relative to minor post-approval changes, where
they function as a signal of bioinequivalence.
The broad spectrum of BA/BE in vitro studies
specifications were provided by each regulatory
authority. 9-18
Statistical Analysis of Bioequivalence:
In the analysis of a bioequivalence study, the
measurements of both Cmax and AUC are
subject to the following procedure. The
measurement for each subject is log transformed
and the averages, µT and µR, of the test and
reference products are calculated. The within
subject variability of the reference product,
σ2WR, is also calculated. There are two parts to
the proposed bioequivalence criteria, a scaled
average bioequivalence evaluation and a point
estimate constraint. In order to demonstrate
bioequivalence both parts must pass. Scaled
average bioequivalence for both AUC and Cmax
128
is evaluated by testing the following null
hypothesis
H0: [(µT- µR) 2/ σ2WR] > Ө
(For given Ө > 0) versus the alternative
hypothesis
H1: [(µT- µR) 2/ σ2WR] ≤Ө
where µT and µR are the averages of the logtransformed measure (Cmax, AUC ) for the test
and reference products, respectively; usually
testing is done at level α=0.05; and Ө is the
scaled average BE limit. Furthermore,
Ө= (ln▲) 2/ σ2Wo
Where ▲ is 1.25, the usual average BE upper
limit for the untransformed test/reference ratio
of geometric means, and σ2Wo =0.25. Note that
rejection of the null hypothesis H0 supports the
conclusion of equivalence.
A 95% upper confidence bound for [(µT- µR) 2/
σ2WR] determined in a BE study must be ≤Ө or
equivalently, a 95% upper confidence bound for
(µT- µR) 2/ Өσ2WR must be ≤0.
Additionally, the point estimate (test/reference
geometric mean ratio) must fall within [0.80,
1.25]. The test drug must pass both conditions
before it is judged bioequivalent to the reference
product. 19
HIGHLY VARIABLE DRUGS AND DRUG
PRODUCTS
In bioequivalence evaluation, highly variable
drugs are generally defined in the context of
within-subject variability in bioequivalence
parameters Cmax and AUC. The most oftenused definition of a highly variable drug is a
drug which has a within-subject (synonymous
with ―intra-subject‖) variability of 30% or more
in these two bioequivalence parameters.
FDA‘s Office of Generic Drugs (OGD)
estimates that approximately 10% of the
submitted BE studies from Abbreviated New
Drug Applications (ANDAs) showed some
evidence of high variability. Examples exist
where a highly variable reference product failed
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Vol. 04 issue 08 April 2012
to demonstrate BE when compared to itself in a
BE study using the standard design/sample size.
As illustrated in Figure 2, because of this high
variability, larger numbers of subjects may be
needed in bioequivalence studies to give
adequate statistical power to meet FDA
bioequivalence limits. The FDA is currently
investigating bioequivalence study design
proposals that can reduce the number of subjects
needed for a bioequivalence study.19-20
HVDs show variable pharmacokinetics as a
result of their inherent properties (e.g.
distribution,
systemic
metabolism
and
elimination). A drug may have low variability if
it is administered intravenously, whereas it can
be highly variable after oral administration. In
such cases, the source of the high variability can
be any of the processes that are involved in the
absorption, such as problematic solubility,
gastrointestinal instability, active transport or
first-pass metabolism in the gut or liver. Davit et
al. recently reviewed 1010 bioequivalence
studies of 180 drugs, of which 31% (57 of 180)
were highly variable.13
About 60% of the surveyed drugs were highly
variable as a result of the pharmacokinetic
characteristics of the drug substances. Several
physicochemical and pharmacokinetic factors
were considered that can potentially contribute
to the observed high variation, such as low
aqueous solubility, acid liability, low
bioavailability (F), pronounced food effect and
so on. Analysis of the data revealed that
extensive first-pass metabolism was probably
the most important factor. Eighty-three percent
of the HVDs were subject to extensive first-pass
metabolism,
whereas the corresponding
proportion in the non-highly variable group was
21%. In addition, the variability may be caused
by the pharmaceutical form in which the drug is
contained. In this case, different formulations of
the same drug may show different within-subject
variabilities (e.g. nadolol). The distinction
129
between HVDs and HVDPs is especially
important with modified release dosage forms
and in formulations of poorly soluble drugs
(Biopharmaceutics Classification System classes
II and IV), where the formulation factors are
more important. Davit et al. related the
variabilities of dissolution performance to those
of bioequivalence parameters. The results
suggested that in about 20% of the highly
variable cases, the performance of drug
formulations could contribute to the high
variation. 13, 20
The factors described above influence
bioequivalence parameter variability due to the
characteristics of the drug substance, rather than
those of the drug product. Drug product
formulation can also contribute to high
variability in bioequivalence parameters. For
example, if the rate of drug release from the
dosage form is highly variable, this factor may
cause high variability in bioequivalence
parameters and may signify a product with lower
product quality. Figure 3 diagrams the steps
involved in bioequivalence evaluation of oral
dosage form performance and illustrate ways in
which high within-subject variability in
bioequivalence measures can arise from either
the drug substance or the drug product.13
Identification of Highly Variable Drugs
The RMSE (Root Mean Square Error) values of
the bioequivalence parameters Cmax and AUC0-t
was used as an estimate of within-subject
variability. Since most of the studies submitted
to the DBE (Division of Bioequivalence) used a
two-way crossover design; it was not possible to
determine the true within-subject variability.
Therefore, the RMSE was used as an estimate of
within-subject variability. Since highly variable
drugs are defined as drugs with within subject
variability of 30% or more in bioequivalence
parameters, we considered a drug to have high
within-subject variability if the RMSE for either
AUC0-t or Cmax was ≥0.3.
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Vol. 04 issue 08 April 2012
Although the FDA evaluates AUC∞ in
bioequivalence studies, but did not define a
highly variable drug as one for which the AUC∞
RMSE≥0.3 because the calculations necessary to
extrapolate to infinity contribute to the
variability of this measure. Therefore, we
consider AUC0-t to be a better indicator of
variability due to drug substance and/or drug
product than AUC∞.
Table 2 shows the number of bioequivalence
studies, drug products, and drugs reviewed by
the DBE in 2003–2005. During this time period,
the DBE found acceptable 1,010 bioequivalence
studies. These 1,010 bioequivalence studies
investigated a total of 524 different drug
products, for 180 different drugs. Frequently,
there are at least several generic versions of any
one reference listed drug under review at the
OGD during the same time period. Each new
generic drug product line is usually the subject
of a separate ANDA. Most ANDAs contain at
least two bioequivalence studies, one under
fasting conditions and one under fed conditions.
A minority of ANDAs contains either one
fasting bioequivalence study or one fed
bioequivalence study. In 111 of these 1,010
acceptable studies, the RMSE was ≥0.3 for
either Cmax and/or AUC0-t. As our criteria for
classification as a highly variable drug was that
the RMSE≥ 0.3 for Cmax and/or AUC0-t, we
concluded that 111 or 11% of these studies were
of drug products that showed high variability in
bioequivalence parameters. These 111 studies of
highly variable drugs were of 101 different drug
products, representing 57 different drugs. 13
Determination of whether high variability in
bioequivalence parameters was consistent
We further classified drugs for which the RMSE
for Cmax and/or AUC0-t≥0.3 as consistently
highly variable, borderline highly variable, or
inconsistently highly variable. Table 3 was
subject to extensive first pass metabolism). As
these are properties of, or factors influencing,
130
the disposition of the drug substance, we
concluded that 61% of the highly variable drugs
reviewed in 2003–2005 were likely highly
variable due to drug substance characteristics.
Notably, several drugs in each of the following
classes were in the consistent and borderline
highly variable groups: angiotensin converting
enzyme (ACE) inhibitors, calcium channel
blockers, 3-hydroxy-3-methylglutaryl-coenzyme
A (HMG-CoA) inhibitors, and bisphosphonates.
All of the ACE inhibitors reviewed during the
2003–2005 period are inactive prodrugs that
undergo extensive first-pass metabolism. The
calcium channel blockers and HMG-CoA
inhibitors reviewed during this period are also
known to undergo extensive first pass
metabolism. The bisphosphonate drugs reviewed
during this period are reported to have absolute
oral bioavailability averaging less than 1%.
Thus, for some potential generic drug products,
it may be possible to predict whether variability
in bioequivalence parameters will be high based
on what is known about the physicochemical
and dispositional characteristics of the drug class
in general. 13
Significance of the HVD Problem;
Although the problem is well known, it is still
very difficult to get hard figures about its extent.
An overview of FDA submissions showed that
about 15% of the applications fell into the
category of HVDs. At first sight, the issue of
HVDs did not appear to be so serious, because
all submitted applications for HVDs passed the
0.80–1.25 regulatory criterion.
However, this regulatory experience was not
shared by other parties involved in generic drug
development. An overview of the database of a
well known Canadian contract research
organization showed a considerably different
picture. In a review of 580 studies, 105 fell into
the highly variable category. The failure rate
was 54%. A very similar figure was reported by
another large Canadian contract research
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Vol. 04 issue 08 April 2012
organization. It appears that only bioequivalence
studies meeting the regulatory expectations were
submitted by applicants and, consequently, the
regulatory agencies could have underestimated
the seriousness of the HVD problem. Some
studies are not undertaken at all when the very
heavy financial and logistical difficulties are
confronted. Diliberti presented a case where the
within subject variation was 173%for the Cmax
and 157%for the AUC at time t (AUCt). He
estimated that at least 598 subjects would be
needed to meet the 0.80–1.25 criterion with 80%
power.
The issue of bioequivalence for hvds is not
only a problem for the generic industry
It has been implicitly assumed until this point
that the single role of bioequivalence studies is
to gain marketing authorization for generic
products. That is not so. For example, it is very
common that a drug formulation used in early
clinical studies is different from that applied in
the late, pivotal investigations. In this case, the
innovator company performs a ‗bridging‘
bioequivalence study in order to demonstrate
that the formulation change does not have a
clinically significant impact. These studies are
typically powered to meet the 0.80–1.25
bioequivalence criterion, partly because of the
convention and partly because at that stage of
product development, there are no firm clinical
safety data.
Also, following Hauschke et al. and Steinijans et
al. the paradigm of bioequivalence is used to
evaluate the drugdrug and drug-food
interactions. In the case of drug interactions, the
lower and upper limits of the bioequivalence
ranges could be different from 0.80 and 1.25,
and alternative ‗effect boundaries‘ could be
allowed on the basis of concentration response
relationships,
pharmacokineticpharmacodynamic models or other available
information.
131
For example, a lack of a food effect is not
considered to be established if the CI is outside
the 0.80–1.25 limits. Altogether, if a new drug
has highly variable features, then to establish
bioequivalence between formulations used in the
product development process or to demonstrate
dose linearity can be a difficult and expensive
challenge. For these reasons, HVDs are not just
a problem for the generic industry but are also a
source of substantial concern to the innovators.
However, compared with generic producers,
regulatory agencies are rather tolerant to the
innovators‘ request for post hoc widening based
on clinical grounds. Because of this, concerns of
innovators about large sample sizes are much
less apparent. 21
Proposals from the Literature
As indicated, the bioequivalence criteria in the
U.S. recommend that the 90% confidence
interval of the geometric mean ratio between the
test and reference products fall within 80-125%.
Over the years, various suggestions have been
made in an attempt to alleviate the difficulty of
meeting the bioequivalence limits for highly
variable drugs and drug products.
Various authors have explored the use of
replicate designs or group-sequential designs. If
a
subject-by-formulation
interaction
is
negligible, the sample size required for a
replicate design study can be reduced up to 50%
of that for a non-replicate design study the
number of study periods is the same since
approximately half the usual number of subjects
is used but they are each studied for twice as
many periods. Therefore, it takes a longer time
to complete a replicate design study, resulting in
an increased chance of subject dropout from the
trial. A group-sequential design may be useful in
cases where there is uncertainty about the
estimates of variability. Nonetheless, the total
number of subjects employed with this design
may be the same as that used for a study without
the group-sequential design if the interim
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Vol. 04 issue 08 April 2012
analysis does not indicate bioequivalence. Also,
to preserve the overall Type I error rate of 5%, a
higher level of confidence interval has to be
used at each stage of the interim analysis.22
Several proposals are available in the literature
to modify the existing bioequivalence criteria for
highly variable drugs and drug products. In
general, these various criteria are based on either
the reduction of the level of the confidence
interval or an increase of the width of the
equivalence limits, or both.
The level of confidence interval reflects the
degree of consumer risk (Type I error in
statistical terms) that can be tolerated by the
regulatory agencies. A reduction in the level of
confidence interval, for example, from 90% to
85%, implies a possible increase in the
consumer risk, which would not be in the best
interests of public health. In contrast, the width
of equivalence limits represents the allowable
boundary for the ratio (or difference) of the
means between products in comparison. Any
adjustment of these limits should be based on
consideration of the statistical properties of the
data as well as on the clinical characteristics of
the individual drug. Statistically, widening the
bioequivalence limits can be accomplished
through expansion of the allowable boundary or
by scaling the criteria based on the high
variability of the reference product. 23-24
PROPOSED SOLUTIONS FOR THE
PROBLEM OF HVD
Relaxation of the Regulatory Requirement:
Health Canada generally expects only that the
point estimate of the GMR (Geometric Mean
Ratio) for the Cmax, but not its 90% CI, should
be between the regulatory limits of 0.80 and
1.25. This relaxed requirement applies generally
and is not aimed specifically at HVDs.
Nevertheless,
it
enables
satisfactory
determination of bioequivalence for several
HVDs because the variation of the Cmax is
usually higher than that of the AUC, and
132
therefore the determination of bioequivalence
can often fail because of the wide CI of the
Cmax. 21
Widening of Bioequivalence Limits Based on
Reference Variability
The bioequivalence limits for these methods are
not determined by the sample size. Rather, they
will be scaled based on the within-subject
variability of the reference product. For both
Methods 2 and 3 below, a side condition to
constrain the mean difference between the test
and reference products has also been proposed.
Method 1:
The rationale for this approach is that a mean
difference of 25% is considered small relative to
the range of values an individual may experience
when the within-subject variability is high, e.g.,
40%. Therefore, the acceptable limits may be
scaled in relation to the size of within-subject
variability as follows:
[U, L] = Exp [σWR]
Where U and L are the upper and lower limits,
respectively; k represents the pth percentile of
the standard normal distribution, Zp; and WR is
the estimated within-subject standard deviation
(obtained from the ANOVA on the log scale) for
the reference. When k = 1, ~ 67% of the
pharmacokinetic measures (such as AUC)
experienced by an individual will be within the
range of [U, L]. Table 4 lists the choices of
limits at k = 1.
Method 2:
A scaled average bioequivalence criterion has
been proposed
[(µT- µR) 2/ σ2WR] ≤Ө
Where µT and µR are the averages of the logtransformed measure for the test and reference
products, respectively; and Ө is the
bioequivalence limit. Comparing Methods 1 and
2, it can be seen that k = Ө -1/2 = (ln1.25)/ σ2Wo
where W0 is the cutoff within-subject standard
deviation for scaling. Relationship of k and σW0
are given in Table 5.
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Method 3:
Derived from the comparison of the distance
measure between the test and reference products,
the following individual bioequivalence criterion
has a reference variance in the denominator, and
thus is scaled to the reference variability.
[(µT- µR) 2 + (σ2WT- σ2WR) + σ2D]/ σ2WR ≤Ө
Where WT is the estimated within-subject
standard deviation for the test product; 2D is the
subject-by-formulation interaction variance
component; and I is individual bioequivalence
limit.
Although theoretically sound, the individual
bioequivalence criterion requires replicate
designs and inclusion of target population in the
study. Because of these resource implications,
the FDA has recommended the continued use of
an average criterion to compare bioavailability
measures.22-27
Direct Expansion of Bioequivalence Limits
Sample size in bioequivalence studies is
determined in large part by the bioavailability
parameter with the highest variability. In most
cases, Cmax has higher variability than AUC.
Thus, widening of the bioequivalence limits for
Cmax has been proposed to reduce the sample
size needed in the evaluation of bioequivalence
for highly variable drugs/products. The greater
variability observed with Cmax may result from
the fact that this parameter is a single point
measurement, which is highly dependent on the
sampling time/frequency and elimination rate of
the drug.
The EMEA currently allows for expanded limits
(e.g., 69.84-143.19%) for Cmax in certain cases
where no safety or efficacy concern arises, based
on the consideration of higher variability for this
measure as compared to AUC.15
Expansion of Bioequivalence Limits Based on
Fixed Sample Size
This method was proposed based on the notion
that only a reasonable number of subjects should
be required for a bioequivalence study.23, 28
133
The number of subjects is fixed by a standard
two-period, crossover study comparing the
reference product with itself where the study
fails to meet the 80-125% limit. The confidence
interval obtained from the reference product in
this study would become the ―goalposts‖ for the
subsequent studies comparing the test with
reference product, using the same number of
subjects.
Expansion of Bioequivalence Limits Based on
Sample Size and Scaling
In addition to fixing the sample size, this method
takes into consideration the producer‘s risk
(Type II error) and reference variability.23 the
equation for the allowable limits is:
[U, L] = Exp [± (tα + tβ/2) n -1/2 σ WR] …….
(Eq.1)
Where α and β are the consumer and producer
risks, respectively; 2n is the number of subjects
desired in the study; and t is the percentile of the
t-distribution with 2n-2 degrees of freedom.
The current regulatory standard has kept the
consumer risk at a level of no more than 5%
while allowing the drug applicant or sponsor to
control its own producer risk. Based on Eq. 1,
for example, assuming a 5% consumer risk and
10% producer risk, the proposed bioequivalence
limits for a typical sample size of 24 subjects
will be
(0.74, 1.35) at σ WR = 0.3
(0.61, 1.65) at σ WR = 0.5
Recent Considerations by Regulatory
Agencies
Although global harmonization is a general goal,
to date, bioequivalence has not been accepted as
a topic by the International Conference on
Harmonization (ICH). Nonetheless, the resource
and ethical concerns for highly variable
drugs/products in bioequivalence are generally
recognized by international regulatory agencies.
It is thus useful to review the differing
regulatory approaches before an informed
recommendation is made on the topic. The
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Vol. 04 issue 08 April 2012
following outlines the bioequivalence standards
used in different regions:
In Canada, for drugs with uncomplicated
characteristics, a 90% confidence limit of 80125% is required for AUC. However, a limit is
placed only on the means (or point estimate) for
Cmax) 11. As a result of random variation or a
larger than expected relative difference, the
sponsor may add more subjects. If this option is
chosen, it must be stated in the study protocol. In
addition, two criteria must be met before
combining is acceptable:
1) The same protocol must be used; and
2) Consistency tests must be met at an alpha
error rate of five percent.
The European Agency for the Evaluation of
Medicinal Products (EMEA) has similar
bioequivalence standards to those in the FDA,
i.e., 90% confidence limits of 80-125% on AUC
and Cmax, with the qualification that these
limits may be expanded in certain cases for
Cmax (e.g., 69.84-143.19%) provided that there
is no safety or efficacy concerns.15
In Japan, the bioequivalence standards also rely
on the 90% confidence limits of 80-125% for
both AUC and Cmax, although wider limits are
allowed for less potent drugs. Additionally, if
the confidence limits are outside of 80-125%,
bioequivalence may be claimed on the grounds
that the study meets. 10 All three conditions
listed below:
1) The total number of subjects in the initial
bioequivalence study is no less than 20
(n=10/group), or pooled sample size of the
initial,
2) The differences in average values of
logarithmic AUC and Cmax between two
products are between log (0.9) – log (1.11); and
3) Dissolution rates of test and reference
products are determined to be equivalent under
all dissolution testing conditions specified.
Japan allows the addition of subjects to increase
the power of a failed bioequivalence study.
134
However, the add-on subjects cannot be less
than half the number in the original study.
South Africa accepts an acceptance interval of
75-133% for Cmax, except for narrow
therapeutic range drugs, when an acceptance
interval of 80-125% applies. For highly variable
drugs, a wider interval or other appropriate
measure may be acceptable, but should be stated
a priori and justified in the protocol.25
Evaluation of Bioequivalence with SABE
Regulatory authorities appear to move towards
adopting the approach of scaled average
bioequivalence (SABE) as a tool for dealing
with the problem of bioequivalence for HV
drugs. Therefore, a brief background of the
procedure will be summarized.
The two one-sided tests procedure is generally
applied for determinations of bioequivalence. In
practice, BE is evaluated by calculating
logarithmic quantities. Thus, means and standard
deviations of the logarithmic data (µ and σ) are
estimated.
Bioequivalence is declared if the difference
between the logarithmic averages is between
limits (BELA) which are preset by regulatory
authorities. Therefore, average bioequivalence
(ABE) is accepted if the following criterion is
satisfied:
- BELA ≤ μT - μR ≤ BELA
The most usually applied regulatory limit is:
BELA = ln (1.25)
(1A)
This assures the earlier stated expectation that
the regulatory limits for the ratio of geometric
means of metrics are 0.80 and 1.25. In practice,
the 90% confidence interval around the
difference between the estimated logarithmic
averages should be between the regulatory
limits.
Thus, regulators need to define, in the case of
average BE, a single criterion for declaring
bioequivalence such as that given in Eq. (1A).
For highly-variable drugs, evaluated by scaled
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Vol. 04 issue 08 April 2012
average BE, two quantities must be defined.
They will be discussed below:
The regulatory criterion suggested for the
application of scaled average BE is:
-BELS ≤ (μT-μR)/σW ≤ BELS
(2)
Here a scaling standard deviation (σW) is related
to the within-subject standard deviation of the
reference formulation (σWR) or, in other views,
is identical to it. This distinction will be
discussed later.
Tothfalusi et al. suggested that the scaled BE
limits (BELS) should be set in the following
form:
BELS = ln (1.25)/σ0
(2A)
Here σ0 is the first measure which should be
defined by regulators. It will be referred to as the
regulatory standardized variation. It defines the
proportionality factor between the logarithmic
BE limits and σW in the highly-variable region
(see Figure 4A). σ0 uniquely determines BELs
and vice versa. For example, when σ0 = 0.294
then BELs is 0.759, and when σ0 = 0.246 then
BELs is 0.907.
Rearranging equation (2), an alternative form is
obtained:
-BELs σW ≤ μT-μR ≤ BELs σ
This form represents average bioequivalence
with expanding limits (ABEL). Consequently,
Eq. 2 and Eq. 2B, i.e. the approaches of SABE
and ABEL, are (almost) identical.
Using the limits of ABEL helps to understand
the properties of SABE from the perspective of
ABE. In this context, the regulatory standardized
variation (σ0) defines the proportionality factor
between the logarithmic ABEL limits and σW
(Figure 1A). A representation of ABEL
conveniently illustrates a mixed regulatory
strategy that was proposed for applying the
unscaled and scaled approaches to the
determination of BE (Figure 4).
According to the mixed regulatory strategy, a
second regulatory term, the so-called switching
variation (CVS), separates regions of low and
135
high variabilities. If the variation of the drug is
low, i.e., when it does not exceed the switching
variation (CVW ≤ CVS) then, following the
present practice, unscaled average BE should be
evaluated. However, for HV drugs when the
variability is higher than the switching variation
(CVW > CVS), scaled average BE is applied.
The mixed regulatory strategy is depicted in
Figure 4 where, for illustrative purposes, SABE
equivalent ABEL limits (BELE* σ ) are plotted.
Two different SABE-equivalent ABEL limits
are shown which correspond to two different
values of σ0. How to set σ0 is the main focus of
this communication.
The standard deviations (σ) can be converted,
approximately, to the corresponding coefficients
of variation:
CV = 100[exp (σ2)-1)] 1/2
(3)
Therefore, for unified and convenient treatment,
the regulatory constants are expressed in terms
of coefficients of variation. As an alternative
notation, CV0 will be used instead of σ0 and the
transformation rule between CV0 and σ0, given
by Eq. 3, will be applied. For example, if σ0 =
0.294 then CV0 = 30%, and when σ0 = 0.246
then CV0 = 25%. The advantage of this unified
notation is that an additional GMR restriction
rule also can be expressed in relative terms. The
0.80-1.25 GMR restriction criterion becomes a
regulatory constraint of 25%. Thus, in our
notation, the proposed mixed approach depends
on three regulatory constants, CVs, CV0 and
CVGMR, with typical values of 30%, 30% and
25%.19-29
Considerations on the implementation of
scaled
average
bioequivalence:
the
recommendations of FDA
As noted earlier, the Advisory Committee for
Pharmaceutical Sciences discussed the topic
repeatedly. At its meeting, on October 6, 2006,
important presentations were offered on behalf
of the FDA Working Group on Highly Variable
Drugs (16-18). The interim recommendations of
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Vol. 04 issue 08 April 2012
FDA were further clarified on May 22, 2007 at
an AAPS/FDA workshop. The current proposals
of FDA and their quantitative characteristics
were published very recently.
FDA has proposed to apply the approach of
reference-scaled average BE for determining the
BE of HV drugs. This means that σW = σWR
would be adopted for scaling.
FDA suggests also that the acceptance criteria
include a constraint on the point estimate for the
ratio of geometric means (GMR). It recommends
that GMR be limited to the range of 0.80 to 1.25.
The Advisory Committee concurred with this
proposal but some members actually favoured a
narrower range.
FDA proposes that both AUC and Cmax should
satisfy the BE acceptance criteria.
FDA recommends that three-period BE studies
be performed in which the reference product (R]
is provided twice and the test product (T) is
given once. Consequently, the possible
sequences of drug administration are TRR, RRT,
and RTR.
The FDA Working Group performed
simulations in order to ascertain the features of
the above proposals. The current FDA
recommendations include a value of σ0 = 0.25.
FDA suggests also that unscaled average BE
used if the within-subject variability is less than
30%, and that reference-scaled average BE
applied if the within-subject variability is at least
30%. These suggestions correspond to a
switching coefficient of variation of CVS =
30%.19-29
Proposed Study Design
For drugs with an expected within-subject
variability of 30% or greater, a BE study with
three-period, reference- replicated, crossover
design with sequences of TRR, RTR, and RRT
is proposed. Specifically, subjects receive a
single dose of the test product once and
reference product twice on separate occasions
with random assignment to the three possible
136
sequences of product administration. This partial
replicate design allows for the estimation of
within subject variability for the reference
product. Treatments should be separated by a
washout period of adequate duration such that
the drug of interest can no longer be detected in
plasma. Subjects recruited for in vivo BE studies
should be 18 years of age or older and capable
of giving informed consent unless otherwise
indicated by a specific guidance. It is the
sponsor‘s responsibility to determine the sample
size needed to achieve the desired power in a
study; however, the minimum number of
subjects that would be acceptable is 24.
The three-period design was selected over a
four-period design because of efficiency. The
only advantage of the four period designs is that
it allows the calculation of the variability of the
test product. The test product variability is not
used in the proposed statistical method. Some
concern has been raised that an ANDA sponsor
may produce a product that has higher
variability than the reference product. However,
under the recommended design, ANDA
sponsors that design a product of lower
variability than the reference product will need a
smaller number of subjects to pass. A
disadvantage of the four-period design is that the
dropout rate for studies increases with the length
of the study. Nevertheless, sponsors may use the
four-period design to demonstrate the BE for
their highly variable drug products.
DISCUSSION AND CONCLUSION
The impact of Cmax variability on the
determination of bioequivalence, as well as the
possible approaches to resolving this issue has
been discussed extensively in the published
literature. Major regulatory agencies have
provisions in their regulations which can
accommodate the effect of higher variability
associated with cmax on the design of
bioequivalence studies. For example, health
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
canada does not require any limits on the
confidence interval for cmax, although limits are
placed on the point estimates for this parameter.
the EMEA and Medicines Control Council of
South Africa both allow for expanded limits for
cmax in certain cases provided that there are no
safeties or efficacy concerns. Similarly, the
Japanese division of drugs accepts limits greater
than 80 – 125%, ―for drugs with
pharmacologically mild actions‖. Additionally, a
failed bioequivalence study can utilize additional
subjects to increase power and the likelihood of
meeting be criteria, provided other conditions
are met.
This report presents a proposal for the BE
evaluation of highly variable drugs and drug
products. This new approach addresses many of
the concerns about the BE of highly variable
drugs/products that have been raised for the past
several years. The proposed approach adjusts the
BE limits of highly variable drugs/products by
scaling to the within subject variability of the
reference product in the study. The
recommendation for the use of reference-scaling
is based on the general concept that reference
variability should be used as an index for setting
the public standard expressed in the BE limit.
Furthermore, for drugs and products that are
highly variable, reference-scaling effectively
decreases the sample size needed for
demonstrating BE. The additional requirement
of a point-estimate constraint will impose a limit
on the difference between the test and reference
means, thereby eliminating the potential that a
test product would enter the market based on a
bioequivalence study with a large mean
difference. The use of the reference-scaling
approach necessitates a study design that
evaluates the reference variability, via multiple
administration of the reference treatment to each
subject. The recommended 3-period design is
the most efficient way to obtain this information.
The proposed approach will resolve a number of
137
issues in the BE evaluation of highly variable
drugs while achieving the FDA‘s mission of
ensuring that all the drugs approved for use in
U.S. are both safe and effective.
ACKNOWLEDGMENTS
I am indebted to my esteemed guide Dr. Anil
Bhandari (Dean, Faculty of Pharmaceutical
Sciences, Jodhpur National University) for his
excellent
guidance,
export
suggestions,
encouragement, support, lively discussion,
constructive criticism, insightful corrections and
everlasting interest in pharmacology and.
1.
2.
3.
4.
5.
6.
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Vol. 04 issue 08 April 2012
Table 1: Brief description about various study designs used in BA/BE studies9-18
Design
Significance
Advantages
Disadvantages
Crossover
• W hen intra-subject CV (approx. 15%) is
usually substantially smaller than that
inter-subject CV (approx. 30%)
• Generally recommended by all regulatory
authorities
• Since the treatments are compared on the same subject, the
inter-subject variability does not contribute to the error
variability
• Subject randomization causes unbiased determination of
treatment effects
• Large information based on minimum sample size
• Straightforward statistical analysis
Parallel
• If the drug has a very long terminal
elimination half-life
• Duration of the washout time for the twoperiod crossover study is so long (if .1
month)
• If the intra-subject CV is higher with
crossover design
Useful for the highly variable drugs (intrasubject CV $ 30%)
• Design is simple and robust
• Drop outs will be comparatively less
• Duration of the study is less than crossover study
• Study with patients is possible
• Straightforward statistical analysis
• Carryover effects and period effects are
possible due to inappropriate wash-out
period
• Long duration
• Possibility of more drop outs leads to
insufficient power
• Not suitable for long half-life drugs
• Not optimal for studies in patients and
highly variable drugs
• Subjects cannot serve as their own controls
for intra-subject comparisons
• Large sample size is required
• Lower statistical power than crossover
• Phenotyping mandatory for drugs showing
polymorphism
Replicate
Variance
balanced
design
• For more than two formulations
• Desirable to estimate the pairwise effects
with the same degree of precision
140
• Allows comparisons of within-subject variances for the test
and reference products
• Indicates whether a test product exhibits higher or lower
within-subject variability in the bioavailability measures when
compared to the reference product
• Provides more information about the intrinsic factors
underlying formulation performance
• Reduces the number of subjects needed in the BE study
• The number of subjects required to demonstrate
bioequivalence can be reduced by up to about 50%
• Design increases the power of the study when the variability
in the systemic exposure of the test drug and formulation is
high
• Allows to choose between two more candidate test
formulations
• Comparison of test formulation with several reference
formulations
• Standard design for the establishment of dose
proportionality
• Involves larger volume of blood withdrawn
from each subject
• Longer duration of the entire study
• Increased possibility of subject drop outs
• Expensive
• Statistical analysis is more complicated
(especially when dropout rate is high)
• May need measures against multiplicity
(increasing the sample size)
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Table 2: Number of Bioequivalence Studies of Highly Variable Drugs Reviewed by the Division of
Bioequivalence in the Office of Generic Drugs from 2003–2005. 13
Description
Bioequivalence Studies
Different Drug Products
Different Drugs
Number
111
% of Total
11
Number
101
% of Total
19
Number
57
% of Total
32
RMSE of AUC0-t
and/or Cmax<0.3
899
89
423
81
123
68
Total number of drugs
studied
1010
100
524
100
180
100
RMSE of AUC0-t
and/or Cmax≥0.3
Table 3: Classification of Variability in Bioequivalence Parameters of Drugs Reviewed by the
Division of Bioequivalence in the Office of Generic Drugs from 2003–2005. 13
Description
Bioequivalence Studies
Different Drug Products
Different Drugs
Number
73
% of Total
66
Number
62
% of Total
61
Number
29
% of Total
51
Borderline
highly
variable drugs
12
11
10
10
6
11
Inconsistently highly
variable drugs
26
23
29
29
22
39
Total
for
which
Cmax and/or AUC0t≥0.3
111
100
101
100
57
100
Consistently highly
variable drugs
141
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Vol. 04 issue 08 April 2012
Table 4: Different k values could be chosen for different drugs depending on their therapeutic
windows.
CV%
30
35
40
45
50
SD(σWR)
0.294
0.340
0.385
0.429
0.472
Lower Limit
0.75
0.71
0.68
0.65
0.62
Upper Limit
1.34
1.40
1.47
1.54
1.60
Table 5: Shows the relationship of k and σW0
142
σW0
k
0.20
1.116
0.223
1.0
0.25
0.893
0.294
0.759
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Figure 1: Brief representation of workflow of bioavailability/bioequivalence study.
143
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Vol. 04 issue 08 April 2012
Figure 2: A visual representation of some possible results of the statistical analyses of
bioequivalence studies. The three bars represent the widths of hypothetical 90% confidence
intervals from bioequivalence studies of drugs with normal variability (green bar), low
variability (blue bar), and high variability (red bar). A bell-shaped curve is superimposed over
green bar, representing the 90% confidence interval, distributed around the geometric mean
test/ reference ratio (―point estimate‖), for the normal variability drug. For simplification, blue
and red bars, respectively, are used in this diagram to represent confidence interval widths of
low variability and highly variable drugs. The blue and red bars also actually represent the
90% confidence intervals of the bioequivalence study Cmax or AUC test/reference ratios
normally distributed about the point estimate. The FDA concludes that a test and reference
product are bioequivalent if the 90% confidence intervals (expressed as a percent) of the
geometric mean Cmax and AUC test/reference ratios fall within the bioequivalence limits of
80–125%. In this illustration, the 90% confidence interval of the normal variability drug
(green bar) meets bioequivalence limits. The 90% confidence interval of the drug with low
variability meets bioequivalence limits although the point estimate deviates from 1.00. For a
highly variable drug, the 90% confidence interval can exceed bioequivalence limits solely
because of the variability. Using more subjects in the bioequivalence study will cause the
90% confidence interval of a highly variable drug to become narrower.20
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Vol. 04 issue 08 April 2012
Figure 3: A diagram relating solid oral dosage form performance to the in vivo system in a
bioequivalence study. Once ingested, a solid oral dosage form disintegrates, then dissolves
into solution (formulation stage). The dissolved drug is absorbed through the gut wall, enters
the liver through the portal vein, and from the liver goes into the systemic circulation, where
pharmacokinetic measurement is possible. From the systemic circulation, the drug reaches the
site of activity from which one observes a clinical response, where pharmacodynamic or
therapeutic measurement is possible. Although the most accurate way of determining
bioequivalence would be to compare test and reference product performance at the
formulation stage, this is nearly always not possible. Consequently, most bioequivalence
studies of systemically absorbed drugs rely on pharmacokinetic measures, as drug blood
concentrations are thought to directly relate to the amount of drug released from the dosage
form. Therefore, a properly designed in vivo study with pharmacokinetic endpoints can
accurately determine whether a test and reference product is bioequivalent. As the drug moves
from the formulation to the systemic circulation to the site of activity, the pharmacokinetic or
pharmacodynamic response becomes increasingly variable with increasing numbers of steps
between the formulation, pharmacokinetic measurement stage, and pharmacodynamic
measurement stage. For example, for drugs that undergo extensive presystemic metabolism,
the effects of the various biotransformation(s) brought about by various gut wall and/or
hepatic metabolism steps contribute to the variability observed in drug pharmacodynamic
measurements. This figure also illustrates the two sources of variability in bioequivalence
measures—variability due to drug substance pharmacokinetics versus variability due to drug
product performance. If high variability exists due to drug substance pharmacokinetics, it may
be necessary to use large numbers of subjects to achieve an acceptable bioequivalence study.
However, if the high variability is due to the formulation or dosage form performance, this
may reflect either a poor quality test or reference product.13
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Vol. 04 issue 08 April 2012
Figure 4: Mixed regul atory model for the determination of bioequivalence. The logarithmic
BE limits, for determinations of average BE with constant and expanding limits, are shown by
thick lines. If the within-subject variation (CVW) does not exceed the switching variation
(CVS) then unscaled average BE is applied, and the BE limits have a constant level of
±log(1.25). When the within-subject variation is higher than the switching variations then the
limits widen with increasing within-subject variation, and scaled average BE can be applied.
The slope (in the logarithmic scale) of the expansion is determined by the regulatory
standardized variation (CV0). The logarithmic average and the SABE-equivalent BE limits
are shown by thick lines. (A) The regulatory standardized variation equals the switching
switching variation, CV0 = 25% and CVS = 30%. The BE limits have a discontinuity at the
switching variation.19-29
146
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
AN
ECONOMIC
ANALYSIS
DIVERSIFICATION IN TAMIL NADU
CROP
V. Kalaiselvi
ijcrr
Vol 04 issue 08
Category: Research
Received on:12/03/12
Revised on:23/03/12
Accepted on:31/03/12
OF
Economics Wing, DDE, Annamalai University, Annamalai Nagar.
Chidambaram,Tamilnadu
E-mail of Corresponding Author: venkimary@yahoo.co.in
ABSTRACT
Crop diversification is considered as a resilience mechanism followed by farmers in different
regions. In the present study, it is shown that there exists crop diversification of crops in
various districts of Tamil Nadu State, India. This is done by constructing a crop
diversification index which provides a basis for ranking the different districts. So in those
regions which are more vulnerable for climatic change, more diversification of crops must be
diversified in order to avoid risk of crop failure and loss of income and employment to the
rural people.
___________________________________________________________________________
INTRODUCTION
A society faced with diminishing natural
resources and every increasing demand for
food consumption and food security due to
increase in population growth, agricultural
intensification is the only course of action
for future growth of agriculture.
Agricultural intensification can be
achieved by changes in cropping pattern or
crop diversification. It is certainly an
important component of the overall
strategy for small farm development. It is
usually viewed as a risk management
strategy. It also provides for self
provisioning in the context of nonmonetized traditional system. As market
opportunities develop and or risks are
somehow reduced, the enterprise mix
begins to respond to market forces and it
was this perspective which was more
relevant in the context of altered economic
environment. Agricultural diversification
really started in the early eighties in India
and it has picked up momentum over the
147
recent past and farmers were always quick
to diversify into higher value crops as
market opportunities developed.
To improve the incomes, to provide
gainful employment and to stabilize the
income flow, diversification of crops
emerges as a major strategy. In several
instances cropping systems have been
diversified or new cropping systems have
been introduced to retain or to enhance the
value of natural resources principally land
and water. There is also the claim that
diversification tends to stabilize farm
income at a higher and higher level. This
happens
when
the
pattern
of
diversification is such as to accommodate
more and more rewarding crops. This is
particularly important for the small
farmers who strive to make their farms,
viable (saleth, 1995).
The study suggested the establishment of
agro
processing
industries
and
infrastructural facilities, arrangement for
crop protection, construction, maintenance
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Vol. 04 issue 08 April 2012
and management of irrigation works,
research prioritization, distribution of
quality seeds and seed materials of the
specific crops in the specific zone on the
basis of cropping pattern and need of the
people of the region.
The study suggested that for achieving the
gains of diversification of farming, there is
an urgent need for further strengthening
the required infrastructure pertaining to
input supply system, marketing system
and the existing research and extension
programmes to increase the adoption of
advanced production technologies.
Saini et al., (1996) in their study on the
impact of diversification on small farms
economy in Kangra district of Himachal
Pradesh observed that the diversification
of arable farming systems with
commercial enterprises such as high
yielding milk animals, poultry birds, beekeeping, floriculture etc, resulted in a
marked increase in the farm income from
6 to 138 per cent. Similarly the capital and
credit requirement showed an increasing
trend with the extent of diversification
implying thereby that to diversify the
existing farming systems with the most
systematically,
remunerative
and
technically feasible enterprises, adequate
facilities should be made available by the
financial institutions.
Given
the
importance
of
crop
diversification under the changing
scenarios a study was undertaken to
examine the crop diversification in
Villupuram District, and to suggest
suitable policy options for furthering the
diversification towards the sustainability
of agriculture in the region.
A main objective of this paper is to
examine
the
patterns
of
crop
diversification at the district level in Tamil
Nadu since 1970-71. There were several
studies relating to the crop diversification
towards commercial crops and most of
148
these were carried out during mid 1990s in
different states of the country. Few studies
on crop diversification were also
conducted in selected district of Tamil
Nadu (Ajjan and Selvaraj, 1996 and
Sunderasan et al., 2002). These evidences
showed that there has been a significant
change in the cropping pattern and a shift
from low value subsistence crop to high
value market oriented crops in Tamil
Nadu. Since the study results reveal the
district wise crop diversification, it will be
useful for district level land use planning
and effective implementation.
This study relied on secondary data, which
were collected from various issues of
Season and Crop Report of Tamil Nadu.
Information on area under 40 crops at
district level for the period between 197071 and 2005-06 were used to analyze the
growth in area, level of diversification and
ranking the districts based on the
diversification.
METHODOLOGY
Measuring Crop Diversification
There are several indices, which explain
either concentration or diversification of
activities in a given time and space by a
single quantitative indicator. Important
indices used to study the corp
diversification are Herfindal Indiex, (HI),
Simpson Index (SI), Ogive Index (OI),
Entropy Index (EI), Modified Entropy
Index (MEI) and Composite Entropy
Index (CEI). Shiyani and Pandya (1998)
and Sundaresan et al., (2002) had applied
more than one of the above indices to
study the diversification of agriculture in
Gujarat and coastal districts of Tamil
Nadu respectively. Joshi et al., (2004)
used Simpson Index of Diversification to
study the patterns of agricultural
diversification in South Asia. Due to the
simplicity in computation and direct
interpretation, the Herfindal index was
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Vol. 04 issue 08 April 2012
employed in this study to examine the
level of diversification. Modified Entropy
index was used to rank the districts based
on the degree of diversification.
The Herfindal Index is a measure of
concentration. It is bounded by Zero and
one: takes a value one when complete
specialization and approaches to zero
when there is diversification of crops. It
was computed as given in equation (1);
sum of squares of the acreage proportion
of each crop in the total cropped area.
Algebraically,
N
Pi 2 .......... .....( 1)
HI
i 1
Where,
Pi
Area under Crop i
Gross Cropped Area
I=1,2….N
N=40 (Number of crops grown int eh
district during the year).
Area under 40 major crops under different
categories (cereals, pulses, oilseeds,
commercial crops, vegetables, fruits,
spices and plantations), total cropped area
in 14 composite districts (Kancheepuram,
South Arcot, North Arcot, Salem,
Dharmapuri,
Coimbatore,
Trichy,
Thanjavur, Pudukottai, Madurai, Ramnad,
Tirunelveli, Nilgris and Kanyakumari) and
state level from 1970-71 to 2005-06 were
used in this study.
RESULTS AND DISCUSSION
Table- 1: Crops selected for the Computation of Diversification Indices
Cereals
Paddy, Sorghum, Pearl Millet, Finger
Millet, Kodo millet, Maize and Foxtail millet
Oilseeds
Groundnut, gingelly, Sunflower Castor and
Niger
Vegetables
Onion, Tomato, Brinjal, Bhendi, Potato,
Tapioca, Sweet potato and Yam
Spices
Chillies, Garlic, Turmeric and Coriander
149
Pulses
Balck gram, Horse gram, Green gram, Red
gram and Bengal gram
Commercial Crops
Sugarcane, Cotton and Tobacco
Fruits
Banana, Mango, Guava and Lemon
Plantations
Cocount, Cardamom, Coffee and Tea.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table- 2
SPREAD INDEX OF CROPS AND CHANGE THE SHARE DURING SELECTED PERIODS IN TAMIL NADU
150
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Table- 3
SPREAD INDEX OF CROPS AND CHANGE THE SHARE DURING SELECTED PERIODS IN TAMIL NADU- (Continued)
151
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Vol. 04 issue 08 April 2012
Table- 4: CROP DIVERSIFICATION AT DISTRICT LEVEL IN TAMIL NADU: HERFINDAL INDEX COEFICENTS
152
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Vol. 04 issue 08 April 2012
Crop Diversification and Ranking the
Districts
Herfindai index is sensitive to the number of
crops grown in the year and their share to the
total cultivated area in the district. Hence,
diversification will not be demonstrated unless
the change in number of crops cultivated in the
year and their share to total cultivated area in the
region is adequately strong to drive the crop
diversification. It is also important to note that
changes in individual crop acreage as well as in
total- cultivated area are taking place
simultaneously, which determine the varying
level of crop diversification for different regions
at different points of time.
Crop agriculture was found highly diversified at
the state level. It was understood from Table 4
the calculated Herfinidal Index coefficient at
State level, showed that the crop diversification
was high (0.16) during 1970-71 and slowly
moving towards diversification (0.13) during the
recent period (2005-06). However, the
diversification level showed variations at the
districts level (Table 2). Among the districts in
Tamil Nadu, Thanjavur, Kancheepuram,
Pudukottai, Kanyakumari and North Arcot
districts exhibited less diversification with the
index coefficients of 0.54, 0.53, 0.31, 0.29 and
0.28 respectively, during the period 1970-71.
Agriculture in Kancheepuram, Kanyakumari and
North Arcot districts became more diversified as
the Herfindal coefficients declined to 0.48, 0.13
and 0.15 respectively for the above districts
during the year 2005-06. Similar trend was
reported by an earlier study conducted by
Sundaresan et al., (2002). But Pudukottai district
has become less diversified which was indicated
by the measure of diversification varying
between 0.24 and 0.38 during the above periods.
Area under minor millets, onion, paddy and
sorghum has declined and there was a significant
growth in area under green gram, sugarcane,
mango and coconut in Kancheerputam distict.
153
Similar pattern was also observed in North Arcot
district. In Kanyakumari district, millets, pulses,
cotton, mango and tapioca witnessed a decline in
area under these crops while coconut and banana
gained their acreage significantly. These
changes were adequately strong to diversify the
crop activities in these districts after the period
1990-91.
Crop activity in Dharmapuri, tirunelveli,
Madurai, Coimbatore, Salem and Trichy districts
was highly diversified and this fact was
supported with the coefficients of 0.08, 0.10,
0.11, 0.12, 0.13 and 0.13 respectively, for the
five years interval starting from 1970-71.
Among the highly diversified districts,
Dharmapuri, Madurai and Salem were moving
towards diversification over the years, while
Coimbatore and Ramanathapuram became less
diversified.
Crop diversification was moderate in south
Arcot presently Cuddalore and Villupuram
Districts and Nilgris districts during 1970-71.
Nilgris district become less diversified
(specialization), which was indicated by
increasing measure of diversification. However,
agriculture in South Arcot has been slowly
diversified over the years. Diversification
pattern in South Arcot was almost similar to that
of Dharmapuri district but the change was
almost similar to that of Dharmapuri district but
the change was adequate to diversify the crop
activities slowly in the district over the years.
Diversification pattern in Nilgris district was
found unique in the State.
Districts like Trichy, Thanjavur and Tirunelveli
were diversified at the same rate over the period
of three decades. Herfindal Index has shown the
pattern and the level of crop diversification at
the district level.
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Vol. 04 issue 08 April 2012
CONCLUSION
Diversification level showed inter- district
variations. For effective planning and
implementation, agricultural development plans
may be designed appropriately for each district
based on the nature and extent of crop
diversification
Though diversifications reduce the risk at farm
level, it would discourage the specialization.
Hence, promotional measures to encourage the
commodity clutters and production efficiency
and necessary. Specialization for instance,
grapes in Theni district, turmeric in Erode
district, mango in (Krishnagiri) Dharmapuri and
Salem districts, maize in Perambalur and
Dindigul districts, Chillies in Ramanathapuram
district, banana in Tiruchirappalli and Tutcorin,
tomato in Dharmapuri district, pepper, tea and
coffee in Nilgris can be promoted.
REFERENCES
1. Ajjan, N. and Selvaraj, K.N. (1996). Crop
Diversification and its Impact in Tamil
Nadu- A Micro Analysis. Indian Journal of
Agricultural Economics, 51 (4):695
2. Arora, V.P.S., Srivastava.S.K (1996).
Divesification of cropping Pattern and
Foodgrain Mix in India: Pace, Magnitude
and Implications. Indian Journal of
Agricultural Economics, 51(4): 699-700.
3. Dhawan K.C., singh, B., Prihar,R.S.,
Brars.S.S.,
and
Arora.B.S
(1996).
Diversification of Indian Agricultural Vis-àvis Food Security. Indian Journal of
Agricultural Economics, 51(4):683-684.
154
4. IFPRI.
(2005)
toward
High-value
Agriculture and Vertical Coordination
Implications
for
Agribusiness
and
Smallholders: Summary of the New Delhi
Symposium, Indian,
5. Joshi, P.K. Ashok Gulati, Pratap S Birthal
and Laxmi Tewari, (2004). Agriculture
Diversification in South Asia. Patterns,
Deteminants and Policy Implciations.
Economic and Political Weekly, 12:24572467.
6. Naik, G. and Jain S.K., (2010). Growth,
Stability and Acreage response of
Agricultural Crops: Likely Impact of WTO
Regime. CMA Monograph No. 191 Oxford
and IBH Publishing Co. Pvt. Ltd., New
Delhi.
7. Shiyani, R.L. and Pandya. H.L. (1998).
Diversification of Agriculture in Gujarat: A
Spatio-Temporal Analysis. Indian Journal of
Agricultural Economics, 53(4): 624-639.
8. Singh, A.J., Jain K.K and Inder Sain. (1986).
Diversification of Punjab Agriculture: An
Econometrical Analysis, Indian Journal of
Agricultural Economics, 41(4): 529-535.
9. Saini, A.S., Sharma K.D., and B.K. Singh
(1996). ―Impact of Diversification on Small
Farms Economy in Himachal Pradesh‖,
Indian Journal of Agricultural Economics,
Vol.51, No.4, pp 697-698.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
OIL-WATER SEPARATION USING FLY ASH ZEOLITE
TREATMENT
Chintan Pathak, V. K. Srivastava
ijcrr
Vol 04 issue 08
Category: Research
Received on:23/03/12
Revised on:27/03/12
Accepted on:02/04/12
Department of Sciences, School of Technology, Pandit Deendayal Petroleum
University, Gandhinagar
E-mail of Corresponding Author: vks1_999@yahoo.com
ABSTRACT
Introduction: A large amount of water is used in the Upstream, Downstream, Petroleum and Automobile
industrial processes and a huge fraction of it comes out as waste after getting polluted by oil and other
toxic substances. Liquid wastes from the Petroleum Industries are relatively less toxic in nature and can
be easily treated by conventional processes. However, solid wastes, especially oily Waste still remains as
major environmental hazards, demanding safer disposal practices. Methodology: Oil contaminated
wastewater is an extremely complex and variable waste of organic compounds ranging up to high
molecular weight tars and bitumen‘s. Oily waste disposal is a major threat to the environment since they
ultimately deplete the natural capital and degrade the prisnity of the eco system. This is the challenging
area for petroleum scientist to establish the method by means of which maximum proportion of oil from
the waste can be recovered. Experimental Set-up: There are different Chemical and Biological
Treatment methods available. The Need is to optimize different treatment technologies in a cost effective
method. The purpose of this work is to increase the oil-water separation by using zeolite treatment along
heat treatment. Result and Conclusion: Results show the oil-water phase separation with zeolite
treatment, as compared to fly ash. Heat treatment may be effectively used to treat oily waste as it gives
more separation in shorter duration of time without creating any environmental as well as human health
hazard. Increase in temperature and duration of heating gives more separation and more reduction in
phase separation.
____________________________________________________________________________________
155
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Keywords: Oil, Oily Waste, Oil Spill, Phase
Separation, Environmental Pollution control
INTRODUCTION
The increasing requirements in the sphere of
environmental protection have induced search for
more effective, inexpensive and ecologically safe
solutions. Given that, the zeolite is aluminosilicate
members of the family - microporous solids
known as ―molecular sieves‖ and have sorptive
and ion-exchange properties; the most important
pollution of hydrocarbon (HC) (viz. petrochemical
spills), may find applications in the removal of oil
spill, using hydrophobic – oleophilic - high-silica
content - hydro thermally stable – zeolite;
particularly those, prepared cheaply from fly ash;
while, simultaneously providing a solution to other
environmental problems. The research will be
applicable in wide variety of environmental
pollution control applications. Since cleanup after
an oil spill is so ineffective and so difficult, and
does not always fully rehabilitate affected areas.
As a challenging task, the author has
experimentally investigated the strategy for
synthesis and application of flyash zeolite for oil
spill cleanup. When we think of oil spills, we
habitually think of oil tankers spilling their cargo
in oceans or seas. However, oil spilled on land
often reaches lakes, rivers, and wetlands, where it
can also cause damage. Oceans and other saltwater
bodies are referred to as marine environments.
Lakes, rivers, and other inland bodies of water are
called freshwater environments. The term aquatic
refers to both marine and freshwater environments.
When oil is spilled into an aquatic environment, it
can harm organisms that live on or around the
water surface and those that live under water.
Spilled oil can also damage parts of the food chain,
including human food resources. The severity of
the impact of an oil spill depends on a variety of
factors, including characteristics of the oil itself.
Natural conditions, such as water temperature and
weather, also influence the behavior of oil in
aquatic environments. Various types of habitats
have differing sensitivities to oil spills as well. The
most interesting results among those described in
156
literature [1-5] on the development and application
of the materials for adsorption purification of
petroleum and oil products from water are
considered and generalized [6].The purpose of
this work is to provide a logical overview of fly
ash synthesized zeolite for oil spill cleanup. Oily
waste disposal is a major threat to the environment
since they ultimately deplete the natural capital
and degrade the prisnity of the eco system.
Potential impacts of oily waste:
 Presence of oily waste in water bodies affects
the potability of water and also its use for
agricultural as well as recreational purposes.
Even traces of oily waste can form a thin film
on the surface of water body and render it unfit
aesthetically. This can also prevent the natural
oxygenation of water bodies and in turn affect
the ecosystem associated with it.
 Layer of oil on aquatic living bodies affects
the metabolic activities.
 Oily waste provides a hostile environment to
the soil bacteria and prevents the natural
nutrient transformation cycle in plants and
microorganisms.
 Oily waste, when exposed to open atmosphere
will disintegrate due to UV rays and will
release volatile organic compounds into
atmosphere
which
are
carcinogenic
compounds in nature. These obnoxious gases
create odour nuisance to the nearby habitat.
There are some treatments given to waste oily
water, shown in Table-1. Sorbents are materials
that soak up liquids. They can be used to recover
oil through the mechanisms of absorption,
adsorption, or both. Absorbents allow oil to
penetrate into pore spaces in the material they are
made of, while adsorbents attract oil to their
surfaces but do not allow it to penetrate into the
material. To be useful in combating oil spills,
sorbent need to be both oleophilic and
hydrophobic (water-repellant).
Although they may be used as the sole cleanup
method in small spills, sorbent are most often used
to remove final traces of oil, or in areas that cannot
be reached by skimmers. Once sorbent have been
used to recover oil, they must be removed from the
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
water and properly disposed of on land or cleaned
for re-use. Any oil that is removed from sorbent
materials must also be properly disposed of or
recycled. Sorbents can be divided into three basic
categories: natural organic, natural inorganic, and
synthetic. Natural organic sorbent include peat
moss, straw, hay, sawdust, ground corncobs,
feathers, and other carbon-based products. They
are relatively inexpensive and usually readily
available. Organic sorbent can soak up from 3 to
15 times their weight in oil, but they do present
some disadvantages [6]. Some organic sorbent
tend to soak up water as well as oil, causing them
to sink. Many organic sorbent are loose particles,
such as sawdust, and are difficult to collect after
they are spread on the water. Adding flotation
devices, such as empty drums attached to sorbent
bales of hay, can help to overcome the sinking
problem, and wrapping loose particles in mesh will
aid in collection. Natural inorganic sorbent include
clay, vermiculite, glass, wool, sand, and volcanic
ash. They can absorb from 4 to 20 times their
weight in oil [7]. Inorganic substances, like
organic substances, are inexpensive and readily
available in large quantities. Synthetic sorbent
include man-made materials that are similar to
plastics, such as polyurethane, polyethylene, and
nylon fibers. Most synthetic sorbent can absorb as
much as 70 times their weight in oil, and some
types can be cleaned and reused several times [8].
Synthetic sorbent that cannot be cleaned after they
are used can present difficulties because they must
be stored temporarily until they can be disposed of
properly.
Adsorbent materials are attractive for some
applications because of the possibility of collection
and complete removal of the oil from the oilspill
site. The further advantage is, it can be, in some
case, be recycled. Some important propertise of
good adsorbent includes, hydrophobicity and
oleophilicity, high updatake capacity, high rate of
uptake, retention over time, oil recovery from it
and reusability. It is interesting to mention that
high-quality zeolite with high water- and oiladsorpotion and CEC may be readily produced
from inexpensive flyash (a by-product of thermal
power stations) and other solid waste materials
157
containing silica and alumina [9-14], thus
providing a solution to other environmental
problems in addition to application in the removal
of oil spills. Zeolite, which are synthesized, show
some of these above mentioned properties. It has
been suggested [10] that zeolite can be useful for
oil spill cleanup due to their oleophilic and
hydrophobic characteristics. In addition, there are
3 properties of zeolite that make them
technologically important: they are selective and
strong adsorbents, they are selective ion
exchangers and they are catalytically active [7].
Researches are increasingly focusing on
hydrophobic pure-silica (or high silica) zeolite as
alternative sorbents for activated charcoal for
sorpotion of organic pollutants (such as volatile
organic compounds) [14].
Hydrophobic zeolite have a small percentage of
aluminum atoms in their crystal structure thereby
shifting their adsorption affinity away from polar
molecules, like water, towards non-polar
substances, like organic solvents (i.e. they are
highly organophillic). These zeolite are thermally
and hydrothermally stable (upto about 1300 oC)
and like other aluminosilicates, they have a unit
structure with a defined pore size of 0.2-0.9 nm,
resulting in a high specific surface ares.
Hydrophobic zeolite also have the advantages like,
little need for safety with regards to fire risk (since
zeolite are inflammable), co-adsorption with water
possible only when the relative humidity is higher
than 70 %, can be regenerated with steam [15-16]
or by calcination at high temperature [17-18]. Thus
the possibility of application of flyash synthesised
zeolite with desired shapes and sizes of are of
great technological value in the oil spill clean-up.
MATERIAL AND METHODS
In order to insure the maximum digestion of silica
from fly ash, it has been opined by the previous
researchers that the microwave can be employed
for complete heating of fly ash samples as
compared to conventional hydrothermal method.
Based on this, the main objectives (viz., less
activation and digestion time, maximum silica
digestion, more nucleation and better crystal
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
growth of zeolite) of the present research has been
planned.
Fly ash samples from Bhusaval Thermal Power
Station (BTPS), Bhusaval, Maharashtra, India;
were collected for the present study. The ash
sampling was done from hoppers and
representative samples were prepared for three
different fields or hoppers. The 40% pure
hydrofluoric acid (HF) from Merck Ltd. was used
for digestion of fly ash by employing microwave.
The most general physical properties of zeolite are
its bulk density and specific gravity (i.e.,
somewhere in between 2 to 2.4) which can
correlate with its porosity (i.e., the measure of the
pore volume in zeolite) and its most dependent
parameter i.e., cation exchange capacity. The type
of zeolite formed is a function of the temperature,
pressure, concentration of the reagent solutions,
pH, process of activation and ageing period, SiO2
and Al2O3 contents of the raw materials.
EXPERIMENTAL SET UP
The samples were collected from the 2 major
sources: [1] Petrol from Petrol Pump (HP) and [2]
Crude Oil from Exploration site. The density
which we have used for sample-1, is 737 kg/m3
and for sample-2, is 825 kg/m3. Fly Ash samples
were collected from Thermal Power Station,
Bhusaval having Density 2-2.3 gm/cc. The Pore
size and surface area of collected fly ash was well
characterized, which is <0.5 nm and >400 m2/g,
respectively. Fly Ash was then treated with 40 %
pure Hydrofluoric Acid (HF) along with
Microwave Irradiation. The range of temperature
was from 45-110 oC. The time range was 5-15
min. The physical characteristics of these
synthesized fly ash zeolites are as follows:
Density: 0.2 to 2.3 gm/cc
Pore sizes: 0.2 to 0.8 nm
Pore volumes: 0.10 to 0.35 cm3/g
Surface areas: 300–700 m2/g
Oil-Water Emulsion was prepared by addition 90
ml of D/W water into 10 ml of Petrol and Crude
Oil samples. It was then kept for 24 hrs. for
formulation of micro-emulsion. The microemulsion which was formed between oil and water
phase, is separated with the help of Seperatory
158
funnel. After obtaining the micro-emulsion, 5 gm
of fly ash was added and treated with the heat
treatment at different temperature ranges from 2545 oC (Fig.- 1,2,3,4; Table-4) for Petrol and Crude
Oil, simultaneously. Similarly, 5 gm of Fly Ash
Synthesized Zeolite were also added into the
micro-emulsion and heat treatment was given at
different ranges from 25-45 oC (Fig.-1,2,3,4;
Table-2 & 3) for Petrol and Crude Oil. Following
parameters were set-up for taking getting result of
Oil-Water Phase Separation using Microwave
Assisted Zeolite Treatment:
Isobaric Specific Heat (kJ/kg.k)
Kinematic Viscosity (cP)
Surface Tension (N/m)
Density (kg/m3)
Separated study of oil and water were also taken
on volume basis and weight basis, after every 5
min.
RESULT
Separation study shows that the fly ash and zeolite,
being the porous material, adsorbing the oil. It is
resulted into the increasing density, from which, it
can be determined that an Emulsion after zeolite
treatment gives increasing in the viscosity and
decreasing surface tension. From this it can be
predicted that as the density increases more
towards water density, the oil-water separation
may take place. From this study of oil-water
separation with the help of fly ash and zeolite, it
has been observed that the separation rate between
oil and water increases with increases in
temperature. After the treatment of flyash with
microwave irradiation into the HF acid, the
adsorption capacity increases because of increase
in pore size.
At 25 oC, the emulsion density is around 829
kg/m3, after zeolite separation the density is 891
kg/m3 and with flyash separation the density is 857
kg/m3. The kinematic viscosity, at 25 oC, of
emulsion is 2.37 cP, with zeolite separation is 12.7
cP and with flyash separation, 5.5 cP. From the
graph-3,4,5,6; it is very clear that almost all the
parameters (viz.,Kinamatic viscosity and Density),
shows the oil-water separation is more favorable
for zeolite as compared to flyash.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
From the graph-1 & 2, it can be seen that the wt. %
up-take of Petrol and Crude Oil, is more using
zeolite compare to flyash. As temperature
increases, it can also be concluded that the Total
Wt. % up-take increases.
4.
5.
6.
CONCLUSION
The microwave-assisted digestion method for fly
ash provided comparable or better results to that of
conventional hydrothermal digestion method. The
variations in weight of sample, temperature, and
time resulted in improved recoveries of several
elements, but generally. In addition, preparation
and time needed for digestion was significantly
reduced with this method.
The microwave-assisted method for fly ash did
work better than hydrothermal digestion method.
There is, however, scope for more improvement in
future research. The use of hydrofluoric acid was
absolutely necessary for digestion of fly ash
because fly ash primarily consists of silicates and
oxides. Most of the elements in ash were
successfully extracted with hydrofluoric acid.
The materials prepared by the method have a pore
structure consisting of micropores as well as
mesopores and macropores. The shape,
composition, pore structure and mechanical
properties of the zeolite microspheres make them
interesting for application in areas such as
adsorption for oil-spill cleanup.
7.
8.
9.
10.
11.
12.
13.
14.
15.
1.
2.
3.
REFERENCES
Metcalf and Eddy, Inc. (1999) ―WasteWater
Engineering‖, 3 rd Edition. 765-915, TATA
McGraw-Hill publishing company Limited,
New Delhi.
Bhatia, S.C. (2002) ―Handbook of Industrial
Pollution and Control‖. 2:312, CBS
publishers.
Cormack, d. (1983) ―Responses to oil and
chemical Marine Pollution‖. Applied science
publishers, New York.
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17.
18.
Noyes, Robert (2005) ―Unit operations in
Environmental Engineering‖. 307, Jaico
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Kiely
uwe
(2006)
―Environmental
Engineering‖. 718, Irwin McGraw-hill.
Noyes, Robert (1993) ―Pollution Prevention
Technology Handbook‖. 30, 39,105,206,502,
Noyes publications, U.S.A.
Sharma, B. K. (2004) ―Industrial Chemistry‖,
14 th edition. Goel publishing house.
Willard, H. H. (1986) ―Industrial Methods of
Analysis‖, 6 th edition. CBS publishers.
Gupta, v. (2006) ―Break through in oil-water
separation‖. Environment science and
Engineering, 57-58.
Rao, M. N. and Datta, A. K. (1978)
―Wastewater Treatment‖, 2 nd edition. Oxford
and IBP publishing Co. Pvt. Ltd.
Punmia, B. C. and Jain, A. K. (2005)
―Wastewater
Engineering‖.
Laxmi
publications.
Gidde, M. R. and Lad, R. K. (2005)
―Environmental Engineering‖, 2nd volume.
Nirali publication.
Chakravarty, R. N. and Bhaskaran, T. R.
(1973) ―Treatment and Disposal of oil
refinery wastes‖, IAWPC volume 10. 137153.
Haruna, J. and Meguro, M. (1992). JP Patent
04012015.
Kuntzel, J.; Ham, R. and Melin, T. (1999).
Chem. Eng. Tech. 22(12), 991.
Kuntzel, J.; Ham, R. and Melin, T. (1999).
Chem. Eng. Tech. 71, 508.
Otten, W.; Gail, E. and Frey, T. (1992).
Chem. Ing. Tech. 64, 915.
Ruthven, D.M. (1988). Chem. Eng. Prog. 84,
42.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table-1: Water treatment methods1
TREATMENT METHODS
1)
2)
FUNCTION
THERMAL REDUCTION
Multiple hearth incineration
Volume reduction & Resource recovery
Fluidized bed incineration
Volume reduction
Wet air oxidation
Volume reduction
Vertical deep well reactor
Stabilization, volume reduction
HEAT DRYING
FLASH DRYER
Spray dryer
Weight and volume reduction
Rotary dryer
Multiple hearth dryer
3)
DEWATERING
VACCUM FILTER
Centrifuge
Volume reduction
Belt filter press
Filter press
4)
THICKENING
GRAVITY THICKENING
Floatation thickening
Centrifugation
V OLUME REDUCTION
Gravity belt thickening
Rotary drum thickening
5)
ULTIMATE DISPOSAL
INCINERATION
Final disposal
Landfill
160
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Vol. 04 issue 08 April 2012
OIL-WATER SEPARATION USING ZEOLITE TREATMENT
(Photo No.1: Oil-Water emulsion)
(Photo No.2: After zeolite treatment)
(Photo No.3: Side view – accumulation on periphery)
161
(Photo No.4: Side view – accumulation on periphery)
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table-2: Petrol-water separation using Zeolite treatment
Zeolite = Density: 2000 to 2300 kg/m3, Pore sizes: 0.2 to 0.8 nm, Pore volumes: 0.10 to 0.35 cm3/g, Surface areas: 300–700 m2/g
Water (D/W) = 90 ml
Sample = 10 ml
Sample
Petrol (HP)
Emulsion (10 % Petrol + 90 % Water)
(Without Zeolite)
Emulsion (10 % Petrol + 90 % Water)
(With Zeolite)
737
786
786
Initial Density (kg/m3)
Temp. (oC)
25
30
35
40
25
30
760
Design Pressure (atm)
35
40
25
30
760
35
40
760
Parameter
Isobaric Specific Heat
(kJ/kg.k)
1.83
1.82
1.81
1.8
1.89
1.79
1.78
1.78
1.74
1.73
1.72
1.72
Kinematic Viscosity
(cP)
2.67
2.83
3.04
3.4
2.37
3.81
4.56
5.79
12.7
13.6
14.5
15.8
Surface Tension (N/m)
0.024
0.0245
0.0251
0.0256
0.0221
0.0261
0.0267
0.0272
0.0296
0.03
0.0304
0.0307
806
814
823
831
829
838
846
853
891
898
905
913
Density (kg/m3)
Time (m)
5
Initial Density of
Zeolite (kg/m3)
2000
2000
2000
2000
Final Density of Zeolite
(kg/m3)
2100
2200
2400
2400
5
10
20
20
Total Up-take
(Wt. %)
162
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Vol. 04 issue 08 April 2012
Table-3: Crude Oil-water separation using Zeolite treatment
Zeolite = Density: 2000 to 2300 kg/m3, Pore sizes: 0.2 to 0.8 nm, Pore volumes: 0.10 to 0.35 cm3/g, Surface areas: 300–700 m2/g
Water (D/W) = 90 ml
Sample = 10 ml
Sample
Crude Oil
Emulsion (10 % Crude Oil + 90 % Water)
(Without Zeolite)
Emulsion (10 % Crude Oil + 90 % Water)
(With Zeolite)
825
840
840
Initial Density (kg/m3)
Temp. (oC)
25
30
35
40
25
30
760
Design Pressure (atm)
35
40
25
30
760
35
40
760
Parameter
Isobaric Specific Heat
(kJ/kg.k)
1.73
1.72
1.72
1.71
1.72
1.71
1.7
1.7
1.7
1.7
1.69
1.69
Kinematic Viscosity
(cP)
63.8
132
132
711
318
420
450
750
388
495
557
862
0.0299
0.0303
0.0303
0.0309
0.0306
0.0309
0.0312
0.0314
0.031
3.13E-02
3.15E-02
3.17E-02
896
903
910
917
911
918
925
932
921
928
935
942
Surface Tension (N/m)
Density (kg/m3)
Time (m)
5
Initial Density of
Zeolite (kg/m3)
2000
2000
2000
2000
Final Density of Zeolite
(kg/m3)
2150
2275
2495
2678
7.5
13.75
24.75
33.9
Total Up-take
(Wt. %)
163
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table-4: Petrol / Crude Oil-water separation using Fly Ash treatment
Fly Ash = Density: 2000 to 2300 kg/m3, Pore sizes: <0.5 nm, Surface areas: >400 m2/g
Water (D/W) = 90 ml
Sample = 10 ml
Sample
Initial
Density
(kg/m3)
Temp. (oC)
Design
Pressure
(atm)
Emulsion (10 % Petrol + 90
% Water) (With Fly Ash)
Emulsion (10 % Crude Oil + 90 % Water)
(With Fly Ash)
786
786
27
164
47
57
27
37
3
47
57
760
760
Isobaric
Specific Heat
1.86
(kJ/kg.k)
Kinematic
Viscosity
5.5
(cP)
Surface
0.00
Tension (N/m) 28
Density
857
(kg/m3)
Time (m)
Initial Density
of Zeolite
2000
(kg/m3)
Final Density
of Zeolite
2060
3
(kg/m )
Total Up-take
(Wt. %)
37
1.84
1.83
1.81
1.71
1.71
1.7
1.69
7.6
8.9
9.9
349
476
497
798
0.02
28
0.029
0.029
0.0307
0.031
0.0313
0.031
865
873
880
913
920
927
934
5
5
2000
2000
2000
2000
2000
2000
2000
2120
2240
2258
2100
2120
2340
2500
6
12
12.9
5
6
17
25
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Graph-1: Petrol Density after Fly Ash & Zeolite treatment
Graph-2: Crude Oil Density after Fly Ash & Zeolite treatment
165
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Vol. 04 issue 08 April 2012
18
Kinematic Viscosity (Petrol)
16
Fly Ash
Zeolite
Emulsion
14
Viscosity (cP)
12
10
8
6
4
2
0
25
30
35
Temp.
40
(oC)
Graph-1,2,3,4
Graph-3: Petrol Kinematic Viscosity after Fly Ash & Zeolite treatment
Graph-4: Crude Oil Kinematic Viscosity after Fly Ash & Zeolite treatment
166
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Graph-5: Wt. % up-take of Petrol using Fly Ash & Zeolite
Graph-6: Wt. % up-take of Crude Oil using Fly Ash & Zeolite
167
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
WIRELESS TECHNOLOGY IN SERVICE OF SOCIETYCASE STUDY OF SNAKEBITE
A
P.P.Patil1, Abhijit A.Patil1, B T Jadhav2
ijcrr
Vol 04 issue 08
Category: Research
Received on:19/03/12
Revised on:24/03/12
Accepted on:30/03/12
1
2
Bharati Vidyapeeth Deemed University, YMIM, Karad
Yashwantrao Chavan Institute of Science, Satara (MS)
E-mail of Corresponding Author: abhijitpatil33@yahoo.com
ABSTRACT
Information communication System plays an important role in the lives of affected Patients of snakebites.
The information systems can be developed which may help to the mankind in emergency by transferring
data. The paper explains the present scenario of Snake bite patients status in the selected sensitive area in
the rural Maharashtra. Survey shows that, the unsatisfactory picture of lack of communication system for
assistance to the affected patients and their further treatment by the hospitals. The paper suggests the
Global Positioning System (GPS) Enabled computing model and the usage of Wireless Technology. The
literature survey shows that, such model may be the need in snake bite affected areas. The Computing
model based on the wireless technology is also useful for making awareness by sharing the information
about the prevention of snake bites and the treatments as first Aid to the community and how does the
data about the victim can be made available and communicated to the further centers such as hospitals.
We mentioned here the advantages and the limitations of the proposed computing model.
The Objective behind the paper is to make use of Information Communication Technology (ICT) to
inculcate the awareness & provide services about prevention of snakebite and treatment directly through
mobile communication system. Method: A prospective analytical study method is used to assess various
risk factors associated with snakebite. Outcome of the study: The paper shows the communication
delays between snakebite patient and hospitals that can be overcome by using wireless technology.
____________________________________________________________________________________
Keywords: GPS, Information System, Wireless
Communication, Computing Model, ICT,
Prevention of Snakebite, Community.
INTRODUCTION
Snakebite is an important and serious problem in
rural Maharashtra. India having 216 species of
snakes out of that only 52 species of snakes are
poisonous [3,5]. The time interval between
snakebite and initiation of treatment is more than
6 hrs. Public Health Care centers limits the
communication and information tools of IT
168
Infrastructure. It is found that there is no proper
reporting system due to underutilization of
information and communication facility to report
about the snakebite victim for immediate
treatment. It has been observed that the
snakebite patients death has been occurred even
in the presence of advanced communication
technology. It is due to absence of emergency
treatment to the patients. Whenever any patient
has suffered by the snakebite in the rural area, it
has been seen that most of the cases are dead
due to lack of communication between patient
and treatment system i.e. hospitals. This paper
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
focus on the use of ICT in snakebite cases in
rural Maharashtra [1]. We observed that the
snakebite cases admitted [2] and referred for
higher hospitals due to the unavailability of
antisnake venom (ASV) that would increase the
chances of uncertainty. The paper suggests a
proposed computing model to help and report
the location and the instant availability of the
antisnake venom.
RESEARCH METHODOLOGY
Snakebite sample cases were collected and
studied from the Department of the Dr
Shankarrao Chavan Government Medical
College and Hospital, Vazirabad, Nanded
District, Maharashtra State, India are shown in
Table 1.
Table 1: Snake bites cases per year from 2000-2010
Year
Snakebite cases
2000
2001
2002
2003
2004
2005
2006
2007
2008
2009
2010
Total
460
570
540
609
645
545
625
603
438
450
427
5997
Death from
snake bite
29
34
33
27
34
25
23
29
14
22
20
290
Percentage (%)
6.30
5.96
6.11
4.43
5.27
4.59
3.68
4.81
3.20
4.89
4.68
53.92
Table 1 shows that, the average death of snakebite patients is 4.90% of total snakebite cases admitted.
Graph 1: Snakebite Patients Vs Death of Patients
169
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table 2: Quarterly Snake bite Incidence in 2000-2010 yrs
Sr. No.
Month (2000-2010)
Total Cases
Percentage(%)
1
Jan-March
450
7.50
2
3
4
April-June
July-Sept.
Oct-Dec.
Total
870
3372
1305
5997
14.51
56.23
21.76
100
Table 2 shows the quarterly snakebite incidence occurred in rural area of Nanded District in Maharashtra,
India
Graph 2: Quarterly Snakebite Incidence
WIRELESS COMPUTING MODEL HELPS
IN TREATMENT OF SNAKE BITES
Internal
Mechanism
of
RAC(Rural
Assistance Center) Computing System:
Incoming call (toll free call) coming from GPS
enabled handset [6,11] to RAC computing
system is handled by GPS Tracking system to
track the location otherwise handled by auto
response machine to capture the name ,address,
query information and store into the database . A
software based knowledge management accept
the general description of snakebite to find out
the type of snakebite and then after further
processing it, display the data about the
antisnake venom [4] and nearest location of the
hospital to get the immediate treatment. This
data is forwarded to the caller, District Health
Office (DHO) and Tahsil Health Office (THO)
170
for further processing to get immediate
treatment to save the life of snakebite victim
Fig. 1 : Ambulance having GPS Tracking
System
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
impact communicated through using ICT based
Wireless Computing Model.
RAC interact with the following Units :
i) Local Self Help Group
ii) Farmers
iii) GramPanchayat
iv) Emergency Services and Ambulance
v) Victims: Affected Users
vi) Local Teachers and Volunteers
vii) District & Tahsil Administration
viii) Research Institutions
Fig. 2 Flow Diagram : Internal working of
RAC (Rural Assistance Center)
Fig 3: Developing awareness among the
population through RAC (Rural Assistance
Center)
Working
Rural Assistance Center (RAC) is one of the
GPS and Web based information cell which
shares the information about snakebite
prevention programmes and awareness amongst
the local users about snakebites and its adverse
171
GPS Enabled RAC comprises a web based
Computing Information Systems which is a
combination of two heterogeneous technologies
viz. Information Communication Technology
(ICT) [8] and Medical Technology (MT). RAC
must work under the control of Regional
Healthcare Center in association with the above
mentioned components.
Roles and Responsibilities of RAC Units
i) Local Self Help Group:
The Local self help group must be trained with
the primary treatment (First Aid) given after the
snakebite immediately. The Local self help
group should contain two senior women
members, two farmers and a snake friend as a
young farmer. The Regional Health Center in
association with research institution and district
and Tahsil administration should provide the
training of this computing system and give the
updated information to the local self help group
through RAC. When any snakebite incident
happens then a member of Local Self Help
Group should be use this system and informed
so that in emergency
ii) Farmers
The farmers should be aware by the RAC with
the help of Gramapanchyat Authority and
Trained Teacher through Awareness Programme
about Prevention of Snake bite and First Aid
Treatment through Web application of RAC.
These trained farmers should convey and share
these information and precautions amongst the
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
community. The RAC should display the
important and Emergency mobile [7] numbers of
close by and local hospitals which provide
treatment of Snake bite affected patients. In
emergency the farmer should contact the above
said emergency numbers and follow their
instructions.
iii) Grampanchyat
The Grampanchyat authority (GRamsevek –
Govt. Officio) should display the pictorial
information prevention and types of snakes and
basic First Aid Treatments through posters.
Conduct awareness Programmes with the help of
School and High Trained Teachers to the
students as well as farmers in the village through
interacting with the RAC system
iv) Emergency Services and Ambulance
(ESA)
Emergency Services and Ambulance (ESA)
should be made available by Civil Surgeon and
Medical Officer through closest Primary Health
Centers. They should display about various
types of Snakes and their Symptoms. They
should have 24x7x366 emergency ambulance
service which will work with RAC computing
system through Wireless Technology [9]. The
Ambulance should be wireless technology based
well equipped with few support of Antisnake
venom. When the Emergency cell receive any
phone call or instruction from RAC computing
system about snakebite patient then it is their
172
responsibility to follow the case through GPS
tracking system and support to the victim so as
to save the life.
v) Victims: Affected Person
Victims or Relatives should just dial the toll free
number of RAC Computing system and follow
the instruction and use the data and information
given by the RAC Computing System.
vi) District & Tahsil Health Administration
District Health Administration has to establish
the Emergency Cell regarding the Snake bite
treatments through Tahsil in association with the
Medical Research Institutions and Private
Hospitals. It is now the Tahsil Health
Administration who should also plan and
execute the instructions received from the
District for the prevention and treatment of
snakebite patients in the villages. RAC
computing system should be controlled by the
Regional
Health
and
District
Health
Administration by providing the updating
medical information related to antisnake venom.
vii) Research Institutions
The Research institutions has to share the
Innovative information about prevention and
First aid treatments, post medical treatments by
making the use of locally available resources
and advanced technology. Research Institutions
must provide the information about advanced
medicines (Antisnake venom) to the RAC and
update it regularly.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Fig. 3 : Interaction between Rural Assistance Center (RAC) units
RESULT
Implementation of RAC Computing will shows
the following results
Immediate reporting of snakebite victim
Quick & accurate information available
about antisnake venom stock and the
location of nearest hospital for treatment
Emergency ambulance and services are
available through this system
Required report generated and Easy to use
Helps to aware about prevention of
snakebite & its treatment
Highly useful to the rural community
The study shows some limitations:
Regular data updating is required
High bandwidth will gives better & faster
results.
DISCUSSION AND CONCLUSION
RAC Computing model is helps to create the
awareness among the rural villagers for
173
prevention & services for snakebite. It has been
found that present manpower in the govt.
hospital studied in this paper is comparatively
less due to which communication gap between
the snakebite victims and hospitals is large. This
deficiency is removed by making use of this
RAC computing model for the needs of rural
society in emergency.
It has been concluded that the RAC computing
model based on wireless technology [10] will be
useful to the snakebite victims and society to
save the life by providing the accurate and faster
information of antisnake venom and nearest
location of the hospital.
ACKNOWLEDGEMENT
We are thankful to the Department of the
Dr.Shankarrao Chavan Government Medical
College and Hospital, Vazirabad, Nanded
District, Maharashtra State, India
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Authors acknowledge the immense help
received from the scholars whose articles are
cited and included in references of this
manuscript. The authors are also grateful to
authors / editors /publishers of all those articles,
journals and books from where the literature for
this article has been reviewed and discussed.
1.
2.
3.
4.
5.
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D. Dalvi [2010]: ― Snake bite : Admissions
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Maharrashtra, India‖, SAMJ, Vol. 100 No. 7
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Virendra C. Patil , Harsha V. Patil , Avinash
Patil, Vaibhav Agrawal ―Clinical Profile and
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International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
PRODUCTION AND OPTIMIZATION OF SINGLE CELL OIL
BY OLEAGINOUS BACTERIA ISOLATED FROM OIL
CONTAMINATED ENVIRONMENTS
T. Murugan1, D. Saravanan1 , R.Balagurunathan2
ijcrr
Vol 04 issue 08
Category: Research
Received on:13/07/11
Revised on:18/08/11
Accepted on:07/03/12
1
PG and Research Department of Microbiology, Sri Sankara arts and Science College,
Tamilnadu
2
Department of Microbiology, Periyar University, Salem, Tamilnadu
E-mail of Corresponding Author: snegapoorvam@yahoo.com
ABSTRACT
Natural sources of oils and fatty acids are derived from plants, animals and microorganism. The oils or
lipids, thus produced from microorganisms are known as ―Single Cell Oil‖. This present study aimed to
isolate lipid accumulating bacteria for the production and optimization of single cell oil. In this
preliminary investigation three soil samples were collected in around Kanchipuram District, 37 different
bacterial strains were isolated and screened by Sudan black stain of that 3 strains (AOM13, AOM17 &
MEW3) showed maximum lipid accumulation. They were used for lipid production of by shake flask
method. The lipid was extracted by Folch method using cell dry matters. Among 3 strains AOM17 was
identified as the efficient producers which produce about 23% of lipid content. It was confirmed as lipid
by solubility, saponification test. Further the lipid was subjected to antimicrobial activity against human
pathogens by well diffusion method, the strain MEW3 showed good activity against Bacillus sp.,
Staphylococcus aureus and Proteus sp. In optimization, all three strains showed maximum lipid
accumulation at 15.5 g/L of glucose, 0.4 g/L of ammonium sulphate, pH 8 and 96 hours incubation time.
The three potential strains AOM13, AOM17 and MEW3 were identified as Cellulomanas sp.,
Arthrobacter sp., Acromicrobium sp., respectively.
____________________________________________________________________________________
Keywords: Single cell oil, Oil production,
oleaginous microbes
INTRODUCTION
Oils, fats and lipids from the natural compounds
are serves as sources of energy and are
considered an important component of our
food3.The demand for oils and fats is largely met
form plant and animal sources 19. Natural
sources of oils and fatty acids are derived from
plants, animals and microorganism9, 19. The
demand for oils and fats in general is largely met
from plants and animals sources3. The main
drawback is, these sources alone cannot be able
175
to meet the total requirement of lipids. The
drawback behind from these sources, a complex
mixture of fatty acids with varying lengths
(Docosahexaenoic acid) and degree of
unsaturation were obtained and it needs
expensive lipid purification. Furthermore, the
fish or animals oil gets contaminated by
environmental factors, which leads to typical
smell and unpleasant taste 19.
Microorganisms will be the suitable alternatives,
as they have the ability to convert a number of
waste materials into a series of valuable-added
products 3. The oils or lipids, thus produced from
microorganisms are known as ―Single cell oil‖
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
and the microbes are called ‗Oleaginous
microbes‘, since they accumulate more than
20% of their biomass as lipids 9. The lipid which
accumulates in oleaginous microorganisms is
mainly
triacylglycerols16.
Oleaginous
microorganism could provide an economically
feasible source of poly unsaturated fatty acids 15.
Some essential fatty acids (EPA) produced by
microorganism are a direct precursor for a
number of biologically active compounds 19 such
as prostaglandins, leukotrienes, thromboxanes
and other related metabolites. It exhibits
regulatory effects on lipoprotein metabolism,
blood rheology, vascular tone, leukocyte
function, plate activation and cell growth 21.
The exploitation of oleaginous microorganism
for the production of single cell oil is value
added products that has relevance and
importance to our nation‘s economy. Most of the
work has so far been, produced single cell oil
mainly from yeasts. Prokaryotic microorganism
should now also consider as a lipids with
potential application in the oil industry. In this
view, the present study was aimed for isolation
and screening for SCO producing bacteria from
different soil samples and to produce single cell
oils in maximum quantity using different carbon
sources based on the optimization of various
factors.
MATERIALS AND METHODS
Collection of Samples
For this study, soil samples were collected from
three different oil contaminated places in around
Kanchipuram District and aseptically transferred
to laboratories for processing.
Isolation and Purification of Bacterial Strains
The samples were serially diluted using sterile
distilled water blanks. Spread plated was made
on Mineral salt agar medium9 and incubated for
24-48
hours
at
room
temperature.
176
Morphologically different bacterial isolates were
selected and purified on Nutrient agar slants and
subsequently analysed for micro morphological
characteristics.
Screening Methods for Single Cell Oil
production
Sudan black B staining: The isolated strains
were stained with Sudan Black B staining 15; 2.
the smear was fixed in the slide and was flooded
with Sudan Black B stain for 15 minutes and
rinsed twice in xylene, followed by counter
stained with diluted safranin for 15 seconds.
Then it was washed with distilled water. The air
dried slide was observed under a phase contrast
microscope on oil immersion for presence of
blue or grayish coloured fat globules within the
cell.
Production of single cell oil
All the Sudan black positive isolates were
studied for the production of lipids. About 2ml
of 24 hours bacterial culture were inoculated in
100 ml of sterilized Supplemented Nutrient
Broth (SNB) medium and placed in a shaker at
200 rpm for 3 to 5 days at 30º C 20.
Cell dry matter
After three days of incubation, the broth was
centrifuged at 5000 rpm for 15 minutes for
separation of pellet. The pellet was washed three
times with sterile distilled water and dried
overnight at 80ºC 8, 9. Then the biomass was
weighed for the determination of cell dry matter
15
Total lipid extraction
Total lipid content of cell dry weight was
extracted by Folch methods 9. The cell dry
matter was extracted with chloroform and
methanol mixture solution (2:1), twice at room
temperature by mixing it for 15-20 minutes in a
shaker and centrifuged at 2000 rpm for 10
minutes. 0.9% Nacl solution was added to the
pellet and vortexed for few seconds, again it was
centrifuged at 2000 rpm for 10 minute. The
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
lower chloroform layer containing lipid was
obtained and allowed for solvents evaporation to
determine the lipid weight 9. Based on the
percentage of the lipid accumulation from the
total cell dry matter 15, different potential
isolates were selected as mentioned in the (Table
2), and the extracted lipids were stored for
further studies.
Confirmation analysis of lipids
Solubility test: A small amount of extracted
lipid from all bacterial isolated were taken in
three test tubes and each of three tubes was
added with water, alcohol, chloroform and
vortexed well to detect the solubility nature.
Saponification test: about 2 ml of 2% NaoH
solution was added to all the small amount of
lipid extracted from potential isolated and mixed
well to emulsify the lipid. Positive result
observed by the formation of soapy solution.
Antibacterial activity of lipids
The single cell oil (lipid) extracted from the
potential isolates were tested for antimicrobial
activity against human bacterial pathogens by
well diffusion method. The 18 hours culture of
Bacillus sp., Staphylococcus sp., Escherichia
coli and Proteus sp., were swabbed on MHA
plates. In each well, the crude lipid (50µl) was
added and incubated at 30º C for 24 hours 7.
Selection of potential strains
Based on the dry weight percentage of total lipid
extracted from the cell dry matter of isolates, the
potential isolates were selected. The isolated
strain was further used for maximum production
of single cell oil and optimization studies.
Optimization of single cell oil production
To enhance production of single cell oil, the
potential strains AOM13, AOM17 and MEW3
were subjected to optimization studies.
Production of SCO was optimized by studying
various nutritional factors (such as carbon and
nitrogen) and environmental (such as pH and
incubation time) was incorporated in Minimal
177
unbalanced agar media 12, 13, 18. To identify the
maximum lipid accumulation in potential
strains, the incubated plates were flooded with
Sudan black staining solution (0.02 % of Sudan
black + 96 % Ethanol) for 30 – 60 minutes and
washed with 96 % ethanol and observed for the
maximum stain absorption 18.
Biosurfactant activity
Hemolysis tests: To observe the hemolytic
activity, the potential isolates were inoculated on
blood agar plates and incubated at 30º C for 72
hours 20.
Emulsification tests: The pure cultures of
potential strains were suspended in test tubes
containing 2ml of Mineral salt solution. After 48
of incubation, 2 ml of kerosene (hydrocarbon)
was added to each tube and the mixtures were
vortexed at high speed for 1 minute and were
allowed to stand for 24 hours to observe
emulsification activity 20.
Characterization of potential isolates
Micro morphological test such as Gram
staining,
Motility,
Spore
staining,
Metachromatic granule staining and biochemical
test such as starch hydrolysis, Gelatin
liquifization, Nitrate reduction and sugar
fermentation tests, etc., were performed to
identify the potential bacterial strains.
RESULTS
Isolation and purification of bacterial strains
A total of 37 morphologically different strains
were isolated from three samples. Among 37
isolates, 17 isolates from soil sample-I, 8 isolates
from soil sample-II and 12 isolates from soil
sample-III were isolated and subcultured.
Among the 37 isolates 35 strains were showed
Gram positive and only 2 strains were showed
Gram negative (Table 1).
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table-1: Micro-morphological characteristics of isolated strains
Strain no.
AOM1
AOM2
AOM3
AOM4
AOM5
AOM6
AOM7
AOM8
AOM9
AOM10
AOM11
AOM12
AOM13
AOM14
AOM15
AOM16
AOM17
MEW1
MEW2
MEW3
MEW4
MEW5
MEW6
MEW7
MEW8
VOM1
VOM2
VOM3
VOM4
VOM5
VOM6
VOM7
VOM8
VOM9
VOM10
VOM11
VOM12
Total
Gram staining
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G- rods
G+ rods
G- rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ rods
G+ cocci
G+ cocci
G+ cocci
G+ cocci
G+ rods
G+ rods
G+ rods
G+ rod
G+rod
G+ rod
35(+)
2(-)
Motility
M
NM
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
NM
NM
M
M
NM
M
NM
M
M
M
M
M
NM
M
NM
M
M
M
31(M) 6(NM)
Catalase
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
37(+)
0 (-)
Oxidase
+
1 (+)
36 (-)
M- Motile; NM= Non motile; + Positive; - Negative
Screening methods for Single Cell Oil
production
Sudan black B staining: Out of 37 isolates, 6
strains were showed positive. 3 strains from soil
sample-I isolate (AOM2, AOM13 & AOM17), 2
strains from soil sample-II isolate (MEW3 &
178
MEW8) and 1 strain from soil sample-III
isolates (VOM12).
Production of single cell oil
The six positive strains namely, AOM2,
AOM13, AOM17, MEW3, MEW8 and VOM12
were used for the production single cell oil. The
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
percentage of total lipid content was calculated
from total cell dry matter and they showed 10%,
18%, 23%, 21%, 16% and 8% respectively
(Table 2).
Table-2: Production of single cell oil
S. No.
Strain no.
Total cell dry matter
(mg/100ml)
Total lipid dry
weight
1
2
3
4
5
6
AOM2
AOM13
AOM17
MEW3
MEW8
VOM12
300
200
325
350
350
740
30
36
75
75
55
60
Total lipid (%
of w/w of
CDM)
10
18
23
21
16
8
compound by the formation of soapy solution
indicating the presence of lipids.
Antibacterial activity of lipids
The crude lipids extracted from potential strain
MEW3 showed antibacterial activity against
Bacillus sp., Staphylococcus sp., and Proteus sp.
(Table 3).
Confirmation for lipids analysis
Solubility test: The extracted lipids were
insoluble in water but soluble in alcohol and
chloroform.
Saponification test: The NaOH solution
saponifies the lipids presented in the extracted
Table 3: Antimicrobial activity of lipids
Strain no.
Bacillus sp.
AOM13
AOM17
MEW3
8
Test organisms (zone of inhibition in mm)
Staph .aureus
E.coli
Proteus sp.
7
7
-
9
Selection of potential strains
Based on the percentage of dry lipid, three isolates namely AOM13, AOM17and MEW3 was selected as
potential strains (Table 4).
Table 4: Potential strains
179
Si. No.
Strain no.
1
2
3
AOM13
AOM17
MEW3
Total cell dry
matter(mg/100ml)
200
325
350
Total lipid dry
weight
36
75
75
Total lipid(% of
w/w of CDM)
18
23
21
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Optimization of single cell oil production
Effect of Nutritional sources
Effect of carbon sources: Growth of all strains
was observed in 4 different concentration of
glucose. All strains showed maximum lipid
accumulation at 1.55 g/L concentration. Strain
AOM13 and MEW3 showed maximum lipid
accumulation at increased concentration of
glucose (1.6g/L).
Effect of Nitrogen sources: In 4 different
concentration of nitrogen source, maximum lipid
accumulation was found at low concentration of
nitrogen souce (0.4g/L). The results were given
in table 5.
Table 5: Effect of Nutritional sources
Strain no.
Growth
(NH4)2
so4
(NSource)
Glucose
(CSource)
Nutritional source
(g/100ml)
1.40
1.50
1.55
1.60
.040
.045
.500
.550
AOM13
Absorption
of stain
Good
Good
Good
Good
Good
Good
Good
Good
+
+++
+++
+++
+++
+++
++
++
Growth
AOM17
Absorption
of stain
Good
Good
Good
Good
Good
Good
Good
Good
++
++
+++
+++
+++
++
+++
++
Growth
MEW3
Absorption
of stain
Good
Good
Good
Good
Good
Good
Good
Good
++
+++
+++
++
++
+++
+++
++
+++ Maximum lipid accumulation; ++ Moderate lipid accumulation and + weak lipid accumulation
Effect of environmental factors
Effect of pH: All strains showed maximum growth at pH 7. Moderate growth was obtained at pH 6 and
poor growth was obtained at pH 5.
Table 6: Effect of pH
Strain no.
AOM13
Absorption
of stain
Poor
Moderate
++
Good
+++
Good
++
Growth
pH
5
6
7
8
Growth
Poor
Good
Good
Good
AOM17
Absorption
of stain
++
+++
+++
MEW3
Absorption
of stain
Poor
+
Moderate
+
Good
+++
Good
+++
Growth
+++ Maximum lipid accumulation; ++ Moderate lipid accumulation; + weak lipid accumulation and - No lipid accumulation
Effect of incubation time: The growth was observed from 24 hours to 96 hours. The increasing hours of
incubation showed strong staining ability which represents the maximum lipid accumulation.
180
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Table 7: Effect of incubation time
Strain no.
Incubation time
24
48
72
96
AOM13
Growth
Absorption
of stain
Good
+
Good
++
Good
++
Good
+++
AOM17
Growth
Absorption
of stain
Poor
Good
++
Good
+++
Good
+++
Growth
Poor
Good
Good
Good
MEW3
Absorption
of stain
+
+++
++
+++ Maximum lipid accumulation; ++ Moderate lipid accumulation; + weak lipid accumulation and - No lipid accumulation
Biosurfactant activity
Hemolysis tests: Among 3 potential isolates, 2
strains AOM17 and MEW3 were showed
hemolytic activity.
Emulsification tests: All the three strains
studied for emulsification activity, which
doesn‘t show any positive result.
Characterization of potential isolates
Based on the cultural characteristics,
microscopic, and biochemical characteristics
(mentioned in Table-8) the three potential
organisms- AOM13, AOM17 and MEW3 were
identified as Cellulomanas sp., Arthrobacter sp.,
Acromicrobium sp., respectively.
Table 8: Characterization of potential strains
Characteristics
Colony morphology
Temperature
Gram staining
Motility
Catalase
Oxidase
Spore staining
Granule staining
Gelatin liquification
Starch hydrolysis
Nitrate reduction
Glucose
Fructose
Maltose
Mannitol
Sucrose
Probable identity
181
Strain-AOM13
Smooth, yellow
pigmented
28˚ C
G+ ve, rods
Motile
+
+
+
+
+
+
Cellulomanas sp.
Strain-AOM17
Dirty white, convex
Strain-MEW3
Dirty white, convex
28˚ C
G+ ve, rods
Motile
+
+
+
+
Arthrobacter sp.
28˚ C
G+ ve, rods
Non-motile
+
+
+
+
+
Acromicrobium sp.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
DISCUSSION
Microbial lipophilic compounds called single
cell oils (SCO), has been the object of research
and industrial interest for many years, due to
their specific characteristics. Such SCO products
are potential for using as alternative sources of
animal or plant oils. According to Gouda et al.,
and Patnayak and Sree 2005 single cell oil is
mainly produced by Fungi and Yeasts, Bacteria
for single cell oil production are not well
explored. In this concern, the present study has
aimed to isolate and identify single cell oil
producing bacteria from different oil
contaminated sites and to optimize for maximum
production.
For the collection of sample, three different oil
contaminated sites were taken and analysed for
isolation of bacteria from oil reservoirs20. In
their present study also totally 37 strains were
isolated from three different oil contaminated
sites. Of this sample-I contained large amount of
microbial load when compared with sample-II
and sample-III. The SCO producers were
screened by sudan black staining and by
determination of total lipid content on cell dry
matter15. In their study also all the 37 strains
were screened by Sudan black stain of that 6
isolates showed positive result for lipid
accumulation. The production of bacterial SCO
was done by shake flask method according to
Gouda et al., 2002.
The lipids were extracted by Foltch method
which is an efficient method to extract the total
lipid content from cell dry matter1, 9, 15. After
extraction, based on the percentage of total lipid
content, 3 isolates were selected. Among 3
strains AOM17 was identified as the efficient
producers which produce about 23% of lipid
content. For the confirmation of extracted
compounds as lipids, lipid analyzing tests were
done and confirmed the presence of lipids. All
182
the 3 strains were taken up for further
optimization studies and application studies.
The lipids extracted from cell dry matter were
analyzed for their antimicrobial properties. In
previous work, Gram positive and yeast only
showed higher sensitivity to the lipid
compounds11 in the present study, the strain
MEW3 showed activity against Gram positive
bacteria and Gram negative bacteria.
The potential strains were optimized for
maximum lipid production and tested
qualitatively by absorption ability of Sudan
black stain. The nutritional factors such as
carbon source and nitrogen source and
environmental factors such as pH and incubation
time were optimized for maximum production of
lipid. This study showed the mild increased
concentration of glucose enhances the
production of lipid. At 15.5 g/L concentration all
three strains AOM13, AOM17 and MEW3
showed maximum lipid accumulation when
glucose (carbon source) used. Papanikolaou et
al. reported 0.5g/L of ammonium sulphate had
produced high lipid. In this study, 0.4 g/L had
showed maximum lipid accumulation when
ammonium sulphate (nitrogen source) used. Hall
and Ratledge, reported that pH had little
influence on lipid accumulation. In this study pH
had influence the accumulation of lipid. Strain
AOM17 and MEW3 showed maximum lipid
accumulation at pH 8 and strain AOM 13
showed maximum lipid accumulation at pH 7.
The effect of incubation time was previously
reported by Papanikolaou et al. In this study, all
the potential strains showed maximum lipid
accumulation at an increased time of incubation.
After 96 hours of incubation, the lipid
accumulation remains constant. The three
potential Single cell oil producers, AOM13,
AOM17 and MEW3 were identified as
Cellulomanas
sp.,
Arthrobacter
sp.,
Acromicrobium sp., respectively.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
CONCLUSION
The present study can be further analyzed by
thin
layer
chromatography
and
Gas
chromatography. In future, many unknown
bacteria may result in the strains that are even
better source of SCO producers than the ones we
know. Efficient production techniques with
detailed knowledge of lipid mechanism should
be developed. This study offers an insight into
exploring the possibility of producing such
valuable added single cell oil at an high amount
and to use it as a supplementary to other edible
fats or to synthesis lipid based products such as
biosurfactant, bioplastics, etc.,
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International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
STUDY OF THE REASONS OF THE RADIOGRAPHIC IMAGES
REPETITION IN SISTAN AND BALUCHESTAN‟S TREATMENT
CENTERS
Mohammad Javad Keikhai Farzaneh1, Reza Afzalipour2, Mojtaba Vardian3,
Mahdi Shirin Shandiz1, Mohammad zarei1
ijcrr
Vol 04 issue 08
Category: Research
Received on:19/01/12
Revised on:01/02/12
Accepted on:25/02/12
1
Zahedan Health Promotion Research Center, Zahedan University of Medical
Sciences, Zahedan, Iran.
2
Medical Physics Dept, Tehran University of Medical Sciences, Tehran, Iran
3
Fasa University of Medical Sciences, Fasa, Iran.
E-mail of Corresponding Author: mojtaba_rmeng@yahoo.com
ABSTRACT
Background and Objective: the repetition of radiographic images causes increased dose of the patients
and personnel, reduced life of the equipment and wasting the national capitals away. By identifying the
percentage of the repetition of radiographic images and the factors associated with it, we can significantly
make help to reducing the dose of patients, increase the useful life of the equipment, increase the
efficiency of personnel, reduce the dose of personnel and make economy in national costs.
Materials and Methods: in this descriptive study, the radiographic images had been collected for 3
months from nine governmental hospitals in the province of Sistan and Baluchestan and also the reasons
why the radiographic images were not accepted by the experts resident in that center was studied. In this
study, the reasons of the repetition of radiographic images were studied as follows: error in exposure
conditions, error in positioning the patient, lack of adoption between radiation center and cast center,
inappropriate choosing of the film size, movement of the patient, the error resulted from radiography
equipment, error in the process of fixation and emergence, lack of appropriate choosing of the radiation
point on the limb and other cases. Findings: Of the 34287 films used in nine treatment center, 4434 films
were repeated and the overall percentages of the repetition of radiographic images were 12.9% which the
maximum percentage of the repetition of radiographic images was in Amiralmomenin Hospital in Zabol
(26.9%) and the minimum percentage was related to Nabiakram Hospital in Zahedan (6.7%). Of the
factors related to the repetition of radiographic images, the maximum percentage of repetition was related
to the high radiation condition (3.22%) and the minimum amount was related to inappropriate choosing of
the film size (0.21%). The percentage of other factors comprised of choosing the radiation condition
(2.38%), error in radiation center (1.88%), error in equipment performance (0.61%), error in the patient‘s
positioning (0.61%), movement of the patient (0.33%), darkroom (1.57%) and other factors (2.08%).
Conclusion : The percentage of the repetition of radiographic images in governmental hospitals in Sistan
and Baluchestan province are in acceptable level in comparison with the statistics issued in other centers,
the percentage of the repetition of radiographic images can be significantly reduced and the national
capitals can be prevented to be wasted away through taking the measures such as regular quality control
of X-ray equipments , training the less experienced personnel and designing the methods of appropriate
choosing of radiation condition.
____________________________________________________________________________________
185
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Keywords: radiography, film, darkroom,
radiation condition, radiographic images, Sistan
and Baluchestan
INTRODUCTION
Due to the fact that X-ray is frequently applied
to diagnose diseases and to reduce the patient‘s
dose and prevent from wasting the national
capitals away, studying the amount and factors
resulted in the repetition of radiographic images
are an unavoidable necessity so that
inappropriate using of X-ray generators causes
the over-exposure of patients and personnel in
radiology department which is contrary to the
ALARA (As Low As Reasonably Achievable)
principle (1).
On the other hand, the repetition of radiographic
images causes increased amount of time in
giving services to patients, the patient‘s
dissatisfaction, reduced useful life of the
equipment and wasting the national capitals
away.
Therefore, the factors which are led to the
repetition of radiographic images in treatment
centers should be taken into consideration and
the re-exposure of the patient should be
prevented as much as possible.
The significant factors of the repetition
radiographic images are as follows: the
inappropriate radiation factors, the error in the
apparatus‘s performance, movement of the
patient, error in the film size, lack of adoption of
the radiation center and cast center, lack of
appropriate adoption of radiation point on the
limb, etc. (2,3). It has been attempted in this
study that the amount of the repetition of
radiographic images are taken into consideration
as well as the most significant factors affecting
on the repetition of images be identified in order
to the appropriate guidelines be presented to
reduce the repetition of radiographic images, and
186
consequently reduce the costs imposed on the
treatment center.
MATERIALS AND METHODS
In this descriptive design, the radiographic
images were collected for three months and have
been studied by observation as such as according
to the overall number of the accepted patients
and the number of applied films, repetition
fraction was calculated according to the equation
below:
Ri=Ai/(Ai+Bi)
Which in this equation:
Ri: fraction of repetition in radiographic center
Ai: total number of repeated films
Bi: total number of accepted films
In the next stage, to determine the factors related
to the repetition of radiographic images, the
form of data collection was designed and some
of the most common factors in the repetition of
radiographic images have been written down as
below:
1- Error in radiation condition which is led to
creating a complete dark or white image and this
error can be investigated having seen the image.
2- Error in patient‘s positioning which can be
investigated by observing the image with
asymmetrical zooming or lack of complete
seeing of the concerned limb.
3- Lack of appropriate choosing of the radiation
point on the limb and lack of adoption of
radiation center with cast center which this error
can be investigated with seeing the image.
4- Inappropriate choosing of the film size.
5- Patient movement, which this error causes the
fading and lack of clarity of the image
6- Error resulted from the equipment which can
be investigated by studying an image with two
projections in a film and seeing the image of the
patient in the experiments which several
radiography are performed to follow up the
performance of the limb.
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
7- The changes in the appearance of the image in
darkroom which this error can be studied by
observing the image, because it can be clearly
seen by inappropriate washing of the film or
dislodging the film‘s emulsion due to the roller‘s
failure.
8- Other factors which can be involved such as
artifact, error in performing the demanded
radiography, etc.
In the next stage radiographic images are
investigated by the experts in radiology in each
hospital and will be involved in the special form
according to the above definition. Finally, after
complete the related forms, the repetition of
radiography images, absolute frequency, relative
frequency and frequency percentage for the
factors related with according to the mentioned
cases are determined and the objectives of the
plan have been realized.
FINDINGS AND DISCUSSION
During performing this design, 34287 films were
used in the concerned hospital. Table 1 shows
the percentage of repetition of radiographic
images in the concerned hospitals.
Table (1): the percentage of repetition radiographic images in the governmental hospitals of
Sistan and Baluchestan province
percentage of repetition films in radiographic
centers
11.8
14.3
6.7
14.3
26.9
12.6
11.8
15.2
7.2
Hospital name
Aliebnabitaleb of zahedan
Boali of zahedan
Nabiakram of zahedan
Taminejtemai of zahedan
Amiralmomenin of zabol
Imamkhomeyni of zabol
Khatam of iranshahr
Iran of iranshahr
Imamali of chabahar
The results achieved in this study indicate that the overall percentage of the repetition films is 12.9%.
The contribution of the percentage of each factors resulted in the repetition for the regarded nine
hospitals has been shown in table 2.
Table (2): the number (percentage) of the repeated images based on the factors resulted in
repetition in governmental hospitals of Sistan and Baluchestan Province
Factors
resulted the
repetition
radiographic
images
repetition
Number
(percentage)
187
High
radiation
condition
low
radiatio
n
conditio
n
Film
size
Patient
positioni
ng
Proces
sor and
darkro
om
Radiatio
n center
Patient
move
ment
Error of
equipment
performance
Other
cases
1105
(3.22)
827
(2.38)
71
(0.21)
208
(0.61)
540
(1.57)
645
(1.88)
114
(0.33)
209
(0.61)
715
(2.08)
International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 08 April 2012
Due to the fact that the range of the percentage of
radiography images repetition in several studies
has been indicated between 0.9% to 27.6% (4-9), it
can be said that the percentage of the repetition in
governmental hospitals of Sistan and Baluchestan
province are in an acceptable range. This study
indicated that high radiation condition and
inappropriate choosing of film size are the most
and least possible causes for repetition,
respectively. Amiralmomenin hospital of zabol and
Nabi -Akram Hospital of Zahedan have the most
and least percentage of repetition, respectively.
In the studies performed by Nixon (7) and Morgan
(9), error in positioning has been indicated as the
most significant cause in the repetition of
radiographic images, while in the current study,
patent positioning and performance error of the
equipment are both in sixth grade.
The admission statistics in Imamkhomeyni of
Zabol and Khatam of iranshahr Hospitals are
higher than other treatment centers and due to the
high lifetime of the equipment of Zabol‘s
Imamkhomeyni Hospital, providing a new
equipment for this hospital is necessary. On the
other hand, inappropriate filtration of the
equipment in Taminejtemai Hospital of Zahedan
causes the percentage of the repetition of
radiography images is high in this center.
Therefore, the filtration function of this equipment
should be corrected. Finally, due to the results
obtained in this study, the following measures can
be effective in reducing the repetition of
radiographic images in governmental hospitals of
Sistan and Baluchestan Hospitals:
Providing the auxiliary equipment to make limit
the regarded limb of the patient, proving the
special charts of choosing the radiation condition
for the employees who are less experienced,
replacing some of the radiography equipments,
regular quality control of the equipments and the
processors and continuous training of the
personnel.
188
The Amiralmomenin Hospital of Zabol also have a
high percentage of exposure repetition which
talking with the patients before being exposed and
the precision of radiologists are necessary to
improve the condition.
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Vol. 04 issue 08 April 2012