A STUDY OF THE IMPACT OF DETERGENT EFFLUENT ON THE
CATFISH MYSTUS GULIO (HAMILTON)
THESIS SUBMITTED TO THE
BHARATHIDASAN UNIVERSITY, TIRUCHIRAPPALLI
FOR THE AWARD OF THE DEGREE OF
DOCTOR OF PHILOSOPHY IN ZOOLOGY
BY
S. KRISHNAVENI, M.Sc., M.Phil., B.Ed., B.L.I.S.,
Assistant Professor of Zoology
A.D.M. College for Women (Autonomus)
Nagappattinam
UNDER THE GUIDANCE OF
Dr. S. RAVEENDRAN, M.Sc., M.Phil., Ph.D., M.Ed., B.L.I.S.,
Associate Professor of Zoology and Research Advisor
POST GRADUATE AND RESEARCH DEPARTMENT OF ZOOLOGY,
KHADIR MOHIDEEN COLLEGE,
(AFFILIATED TO BHARATHIDASAN UNIVERSITY)
ADIRAMPATTINAM – 614 701
TAMIL NADU, INDIA
OCTOBER - 2012
Dr. S. RAVEENDRAN M.Sc., M.Phil., Ph.D., M.Ed., B.L.I.S., Office : 04373-242236
Associate Professor of Zoology,
Resi : 04373-235585
P.G. and Research Department of Zoology,
Mobile : 9443617904
Khadir Mohideen College,
Email: drsr_kmc@rediffmail.com
Adirampattinam – 614 701.
CERTIFICATE
This is to certify that the thesis entitled “A STUDY OF THE IMPACT OF
DETERGENT EFFLUENT ON THE CATFISH MYSTUS GULIO (HAMILTON)”
submitted to Bharathidasan University, Tiruchirappalli by S. KRISHNAVENI in partial
fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY IN
ZOOLOGY, is a record of research work done by her during the period of study under
my guidance and supervision. I further certify that the thesis has not previously formed
the basis for the award of any degree, diploma, associateship, fellowship or other similar
title.
Research Guide and Supervisor
Station : Adirampattinam
Date :
DECLARATION
I do hereby declare that this work has been originally carried out by me
under the guidance and supervision of Dr. S. RAVEENDRAN, M.Sc., M.Phil.,
Ph.D., M.Ed., B.L.I.S., Associate Professor of Zoology, P.G. and Research
Department of Zoology, Khadir Mohideen College, Adirampattinam, and that this
work has not been submitted elsewhere for any other degree.
(S. KRISHNAVENI)
Station: Adirampattinam
Date :
ACKNOWLEDGEMENT
I would like to express my heartfelt indebtedness and warm regards to my
research mentor Dr. S. Raveendran, M.Sc., M.Phil., Ph.D., M.Ed., B.L.I.S., Associate
Professor of Zoology, P.G. and Research Department of Zoology, Khadir Mohideen
College, Adirampattinam, for suggesting the problem, continuous encouragement, able
guidance at every stage of my work and also for introducing me to this enlightening field
of research and started me on the path that led to this Ph.D. Thesis.
I express my respectful and sincerest gratitude to Dr. A. Mohamed Abdul
Kader, Principal, Khadir Mohideen College, Adirampattinam for the facilities provided
to complete the work
I am grateful to Dr. P. Kumarasamy, Associate Professor & Head, P.G. and
Research Department of Zoology, Khadir Mohideen College, Adirampattinam, for his
encouragement and valuable suggestions.
I am highly indebted to Dr. A. Amsath, Associate Professor of Zoology,
Dr. Muthukumaravel, Dr. O.Sathick, Mrs. V. Ganapriya, Dr. A. Maharajan and.
Dr. J. Sukumaran Assistant Professors of Zoology, P.G. and Research Department of
Zoology, Khadir Mohideen College, Adirampattinam, for their valuable suggestions and
help in finishing the thesis.
I
have
great
pleasure
in
expressing
my
sincere
thanks
to
Dr. G. Sridharan, M.Sc., M.Phil., Ph.D., Associate Professor, P.G. and Research
Department of Zoology, Rajah Serfoji Government College (Autonomous), Thanjavur,
for providing me all the necessary help to complete my this work.
I am immensely grateful to Dr. V. Ramamurthy, M.Sc., M.Phil., Ph.D., M.L.T.,
Assistant Professors of Biochemistry, Marudupandiyar College, Thanjavur, for his
valuable guidance and suggestions through out the tenure of this work. I acknowledge his
kind help and constant encouragement with a deep sense of gratitude.
I would like to express my special thanks to The Secretary and Members of
Management of A.D.M College for women (Autonomous), Nagappattinam, for their
advice and encouragement throughout the course of my research work and providing
official permission given to constant support be extended during the course of my
research work
My respect and appreciation go to Dr. Mrs. A. Sivakamasundari, Principal,
A.D.M College for women (Autonomous), Nagappattinam, for her guidance, continual
support and advice throughout my study.
I am thankful to Dr. Mrs. V. Valarmathi, Head, Department of Zoology, A.D.M
College for women (Autonomous), Nagappattinam for her valuable suggestions and
guidance to conduct and complete the research work.
I would like to thank Dr. V. Renuga, Controller of Examinations A.D.M College
for women (Autonomous), Nagappattinam, for her encouragement and constant support
during the course of my research work.
It is indeed a pleasure to acknowledge the timely help extended by our colleague
of Biochemistry and Zoology departmental faculty members who have generously
spared their precious time and made their contributions to this work.
I shall be failing in my duty, if I do not acknowledge the moral support of my
family members. This work would not have been possible without the enthusiasm,
support and encouragement of my nephew J. Yoheshwaran, who has extended
enthusiastically is worthy to be mentioned here to make this acknowledgement complete
I thank to Mr. P. Muruganantham, Murugan Binding Works, Pattukkottai, for
his neet typing and binding for this project work successfully.
There are times when words are inadequate to express the profound sense of
gratitude and appreciation one feels in the heart. “Praises can be had only from a
grateful heart”. I grant my grateful appreciation and thanks to all those who have
directly and indirectly contributed to this work.
(S. KRISHNAVENI)
LIST OF TABLES
Table 1. Characteristics of detergent effluent
Table 2. Biochemical characteristics of isolated bacteria
Table 3. Bacterial flora from detergent effluent
Table 4. Fungal flora from detergent effluent
Table 5. Cyanobacterial flora from detergent effluent
Table 6. Effect of different concentration of detergent effluent on mortality percentage of
M. gulio during 24, 48, 72, 96 Hrs. of exposure
Table 7. Effect of detergent effluent on surfacing behaviour of Mystus gulio
Table 8. Effect of detergent effluent on Opercular beats of Mystus gulio
Table 9. Effect of detergent effluent on rate of oxygen consumption of
Mystus gulio
Table
10.Haematological parameters of Mystus gulio
exposure to sublethal concentration of detergent effluent
after
10
days
Table
11.Haematological parameters of Mystus gulio
exposure to sublethal concentration of detergent effluent
after
20
days
Table
12.Haematological parameters of Mystus gulio
exposure to sublethal concentration of detergent effluent
after
30
days
Table 13. Total protein content in the muscles and liver of Mystus gulio exposed to
detergent effluent
Table 14. Glycogen content in the muscles and liver of Mystus gulio exposed to detergent
effluent.
Table 15. Amino acids content in the muscles and liver of Mystus gulio exposed to
detergent effluent
Table 16. Lipid content in the muscle and liver of Mystus gulio exposed to detergent
effluent
Table 17. Levels of LDH activity in the muscles and liver of Mystus gulio exposed to
detergent effluent
Table 18. Levels of SDH activity in the muscles and liver of Mystus gulio exposed to
detergent effluent
Table 19. GOT content in the muscles and liver of Mystus gulio exposed to detergent
effluent
Table 20. GPT content in the muscles and liver of Mystus gulio exposed to detergent
effluent
Table 21. ALP content in the muscles and liver of Mystus gulio exposed to detergent
effluent
Table 22. ACP content in the muscles and liver of Mystus gulio exposed to detergent
effluent
Table 23. Phosphatase and protease content in the tissues of Mystus gulio exposed to
detergent effluent
LIST OF FIGURES
Fig. 1. Experimental animal Mystus gulio
Fig. 2. A natural habitation of Mystus gulio
Fig. 3. Section in the gill of a control fish of Mystus gulio (Staining Haematoxylin /
Eosin Magnification 320 X)
Fig. 4. Section in the gill of 0.2% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 5. Section in the gill of 0.4% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 6. Section in the gill of 0.6% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 7. Section in the gill of 0.8% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 8. Section in the liver of a control fish of Mystus gulio (Staining Haematoxylin/
Eosin Magnification 320 X)
Fig. 9. Section in the liver of 0.2% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 10. Section in the liver of 0.4% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 11. Section in the liver of 0.6% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 12. Section in the liver of 0.8% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 13. Section in the kidney of a control fish of Mystus gulio (Staining Haematoxylin /
Eosin Magnification 320 X)
Fig. 14. Section in the kidney of 0.2% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 15. Section in the kidney of 0.4% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 16. Section in the kidney of 0.6% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 17. Section in the kidney of 0.8% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 18. Section in the intestine of a control fish of Mystus gulio (Staining Haematoxylin /
Eosin Magnification 320 X)
Fig. 19. Section in the intestine of 0.2% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 20. Section in the intestine of 0.4% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 21. Section in the intestine of 0.6% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 22. Section in the intestine of 0.8% of detergent effluent treated fish of Mystus gulio
(Staining Haematoxylin / Eosin Magnification 320 X)
Fig. 23. Bar diagram showing the temperature of detergent effluent
Fig. 24. Bar diagram showing the pH of detergent effluent
Fig. 25. Bar diagram showing the total suspended solids of detergent effluent
Fig. 26. Bar diagram showing the total dissolved solids of detergent effluent
Fig. 27. Bar diagram showing the free carbon dioxide of detergent effluent
Fig. 28. Bar diagram showing the carbonate of detergent effluent
Fig. 29. Bar diagram showing the bicarbonate of detergent effluent
Fig. 30. Bar diagram showing the BOD of detergent effluent
Fig. 31. Bar diagram showing the COD of detergent effluent
Fig. 32. Bar diagram showing the Dissolved oxygen of detergent effluent
Fig. 33. Bar diagram showing the Nitrate of detergent effluent
Fig. 34. Bar diagram showing the Nitrite of detergent effluent
Fig. 35. Bar diagram showing the Ammonia of detergent effluent
Fig. 36. Bar diagram showing the total phosphate of detergent effluent
Fig. 37. Bar diagram showing the inorganic phosphate of detergent effluent
Fig. 38. Bar diagram showing the organic phosphate of detergent effluent
Fig. 39. Bar diagram showing the calcium of detergent effluent
Fig. 40. Bar diagram showing the magnesium of detergent effluent
Fig. 41. Bar diagram showing the chloride of detergent effluent
Fig. 42. Bar diagram showing the sublethal effects of detergent effluent on the number of
visits of M.gulio
Fig. 43. Bar diagram showing the sublethal effects of detergent effluent on the opercular
beats of M.gulio
Fig. 44. Bar diagram showing the sublethal effects of detergent effluent on the Oxygen
consumption of M.gulio
Fig. 45. Bar diagram showing the RBC number changes in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 46. Bar diagram showing the WBC number changes in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 47. Bar diagram showing the Hb changes in M.gulio with the effects of detergent
effluent exposed to different days and different concentrations
Fig. 48. Bar diagram showing the PCV changes in M.gulio with the effects of detergent
effluent exposed to different days and different concentrations
Fig. 49. Bar diagram showing the MCV changes in M.gulio with the effects of detergent
effluent exposed to different days and different concentrations
Fig. 50. Bar diagram showing the MCH changes in M.gulio with the effects of detergent
effluent exposed to different days and different concentrations
Fig. 51. Bar diagram showing the MCHC changes in M.gulio with the effects of detergent
effluent exposed to different days and different concentrations
Fig. 52. Bar diagram showing the protein content of muscles in M.gulio with the effects
of detergent effluent exposed to different days and different concentrations
Fig. 53. Bar diagram showing the protein content of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 54. Bar diagram showing the glycogen content of muscles in M.gulio with the effects
of detergent effluent exposed to different days and different concentrations
Fig. 55. Bar diagram showing the glycogen content of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 56. Bar diagram showing the amino acids content of muscles in M.gulio with the
effects of detergent effluent exposed to different days and different
concentrations
Fig. 57. Bar diagram showing the amino acids content of liver in M.gulio with the effects
of detergent effluent exposed to different days and different concentrations
Fig. 58. Bar diagram showing the lipid content of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 59. Bar diagram showing the lipid content of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 60. Bar diagram showing the LDH activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 61. Bar diagram showing the LDH activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 62. Bar diagram showing the SDH activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 63. Bar diagram showing the SDH activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 64. Bar diagram showing the GOT activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 65. Bar diagram showing the GOT activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 66. Bar diagram showing the GPT activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 67. Bar diagram showing the GPT activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 68. Bar diagram showing the ALP activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 69. Bar diagram showing the ALP activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 70. Bar diagram showing the ACP activity of muscles in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 71. Bar diagram showing the ACP activity of liver in M.gulio with the effects of
detergent effluent exposed to different days and different concentrations
Fig. 72. Bar diagram showing the phosphatase activity of tissues in M.gulio with the
effects of detergent effluent exposed to different days and different
concentrations
Fig. 73. Bar diagram showing the protease activity of tissues in M.gulio with the effects
of detergent effluent exposed to different days and different concentrations
CONTENTS
CHAPTER
TITLES
PAGE NO
INTRODUCTION
1
Objectives of the Study
21
2
REVIEW OF LITERATURE
23
3
MATERIALS AND
51
1
1.1
METHODS
Experimental Animal
51
3.1.1
Collections and maintenance of fish
51
3.1.2
Experimental setup
51
Effluents
52
3.2.2
Methods of effluent analysis
52
3.2.3
Biodiversity of microbes
53
3.2.4
Survival Studies
56
3.2.5
Experimental condition
57
Behaviour Studies
57
3.3.1
Recording of surfacing behaviour
57
3.4
Respiratory Behaviour Studies
57
3.4.1
Recording of opercular movements
57
3.4.2
Rate of Oxygen consumption
58
Heamotological Studies
58
Erythrocytes
58
3.1
3.2
3.3
3.5
3.5.1
3.5.2
Total Leucocytes
59
3.5.3
Haemoglobin content
59
3.5.4
Packed Cell Volume (PCV)
59
3.6
Biochemical Analysis
60
3.6.1
Estimation of proteins
60
3.6.2
Quantitative Estimation of Glycogen
62
3.6.3
Estimation of free Amino acids
63
3.6.4
Estimation of Lipids
63
Enzyme Assays
64
3.7.1
Determination of the Activity of GPT
64
3.7.2
Determination of the Activity of GOT
65
3.7.3
Estimation of Alkaline Phosphatase
66
3.7.4
Estimation of Acid Phosphatase
67
3.7.5
Estimation of Phosphatase
68
3.7.6
Estimation of Protease
69
3.7.7
Lactate Dehydrogenase activity
70
3.7.8
Succinate Dehydrogenase activity
70
Histology - Paraffin methods
71
3.7
3.9
4
72
RESULTS
4.1
Physico-chemical
characteristics
of
72
detergent effluent
4.2
Biodiversity of microbes in effluent
72
4.2.1
Bacteria
72
4.2.2
Fungi
73
4.2.3
Cyanobacteria
73
4.3
Effects of detergent effluent on Fish
74
4.4
Behaviour changes of Mytus gulio
75
4.4.1
Swimming activity
75
4.4.2
Surfacing activity
75
4.5
Respiratory behaviour
76
4.5.1
Opercular movements
76
4.5.2
Rate of oxygen consumption
76
4.6
Haematological parameters
77
4.6.1
10th day – exposure
77
4.6.2
20th day – exposure
78
4.6.3
30th day – exposure
78
Biochemical Studies of Mystus gulio
79
4.7.1
Total protein content in muscle
79
4.7.2
Total protein content in Liver
79
4.7.3
Glycogen content in muscle
80
4.7.4
Glycogen content in liver
80
4.7.5
Free amino acids in muscle
81
4.7.6
Free amino acids in Liver
81
4.7.7
Lipid content in muscle
82
4.7.8
Lipid content in liver
82
Enzymes Activities of Mystus gulio
82
4.8.1
LDH Activity in muscles
82
4.8.2
LDH Activity in Liver
83
4.8.3
SDH Activity in muscles
83
4.8.4
SDH Activity in Liver
84
4.8.5
GOT and GPT activity
84
4.8.6
Alkaline phosphatase activity
85
4.8.7
Acid phosphatase activity
86
4.8.8
Phosphatase Activity
87
4.8.9
Protease Activity
87
Histopathlogical Studies
87
4.9.1
Histology of Gills
87
4.9.2
Histology of Liver
89
4.9.3
Histology of Kidney
90
4.9.4
Histology of Intestine
91
4.7
4.8
4.9
5
147
DISCUSSION
5.1
Physico-chemical
characteristics
of
147
detergent effluent
5.2
Biodiversity of microbes
151
5.3
Survival studies of Mystus gulio
155
5.4
Behaviour Studies of Mystus gulio
157
5.5
Haematological Studies of Mystus gulio
162
5.6
Biochemical Studies of Mystus gulio
165
5.7
Enzymes Studies of Mystus gulio
175
5.8
Histological studies of Mystus gulio
185
6
SUMMARY
199
7
REFERENCES
207
APPENDIX