BIOFILM INHIBITORY ACTIVITY OF
BIOSURFACTANTS ISOLATED FROM THE
MARINE BACTERIA
By
Pandi selvi, K.
Reg.No:09MBT12
Under the guidance of
Dr. S. SATHEESH
Lecturer
International Centre for Nanobiotechnology
Manonmaniam Sundaranar University
Rajakkamangalam – 629 502
Kanyakumari District
Tamil Nadu, India
Synopsis
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Introduction
Objectives
Materials and Methods
Results and Discussion
Summary
Acknowledgement
References
INTRODUCTION
• Biofilm is an assemblage of microbial cells that is
irreversibly associated with a surface and enclosed in a
matrix of primaroly polysaccharide material.
• Biofilm form when bacteria adhere to surfaces and begin
to excrete a glue-like substance that protects and
anchors them to material such as metals such as
metals, plastic, soil, medical implants and tissue.
Cont…
• In industry, biofilms can cause corrosion of pipes,
reduce heat transfer in cooling systems and plug water
filters.
• The major difficulties in controlling surface-attached
bacteria is their enhanced resistance to antimicrobial
agents.
• Surface active compounds (surfactants) have been
empolyed to prevent bacterial adhesion to surfaces.
CONTROL OF BIOFILM
• A biofilm control programme is detection and
assessment of various biofilm components like thickness
of slime layer, algal and bacterial species involved,
extent of extracellular polymeric substances (EPS) and
inorganic components.
• The new techniques for biofilm control like ultrasound,
electrical fields, hydrolysis of EPS and methods altering
biofilm adhsion.
BIOSURFACTANT
• Surfactants both chemical and biological are amphiphilic
compounds which can reduce surface and interfacial
tensions by accumulating at the interface of immiscible
fluids.
• It play an essential role in the swarming motility of
microorganisms and participate in cellular &
physiological processes of signalling and differentiation
as well as in the biofilm formation.
Cont…
• There are several advantages over the chemical
surfactants such as lower toxicity, higher
biodegradability, better environmental compatibility,
higher foaming , high selectivity and specific activity at
extreme temperatures, pH and salinity and the ability to
synthsized from renewable feedstock.
OBJECTIVES
• to isolate and characterize the biosurfactant from
seaweeds and gastropods,
• to check the antibacterial activity,
• to analyse the impact of biosurfactant on bacterial
adhesion,
• to characterize the biosurfactant by TLC, HPLC & NMR.
MATERIALS AND METHODS
• Isolation of surface associated bacteria (SAB),
• Biochemical characterization of isolated bacterial strains,
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Gram’s staining
Motility test
Indole production test
Methyl-red test
Voges-proskauer test
Citrate utilization test
Starch hydrolysis
Urea hydrolysis
Fermentation of carbohydrate
Catalase test
Gelatin hydrolysis test
Cont…
• Isolation of biosurfactant (Oil displacement test),
• Antibacterial activity of biosurfactant against biofilm
bacteria,
• Influence of biosurfactants on the adhesion of biofilm
bacteria to surface,
• Characterization of biosurfactant by TLC, HPLC & NMR.
Results and Discussion
+ - Positive
•
- Negative
S1 - Seaweeds
S2 - Seaweeds
S3 - Seaweeds
S4 - Seaweeds
G1 - Gastropods
Biosurfactant screening methods
Antimicrobial activity of biosurfactant isolates from the srain
S1, S2 against target bacteria
Antimicrobial activity of biosurfactant isolated from the
strain
S 3, S4 & G1.
Adhesion assay using biosurfactant of the strain S1 & S 3
(number of bacterial cells adhered on glass coupons)
Thin layer chromatogram of the biosurfactant isolated from
the strain S1, S2, S3, S4 & G1.
HP LC spectrum of biosurfactant isolated from the strain
S1
H P LC spectrum of biosurfactant isolated from the strain S 3
H P LC spectrum of biosurfactant isolated from the strain S4
1
H NMR spectrum of the biosurfactant isolated from the strain S 3
13
C NMR spectrum of the biosurfactant isolated from the strain S3
DISCUSSION
• In conclusion, result of the present study revealed that
bacteria associated with marine invertebrates and
seaweeds has the potential to produce biosurfactants.
• Further studies on this aspect may improve our
knowledge on the application of these biosurfactants in
biomedical and environmental field.
SUMMARY
• The present study was carried out to assess the biofilm
inhibitory activity of bacteria associated with seaweeds
and gastropods.
• Biosurfactant was tested for antimicrobial activity against
five bacterial strains (E.coli, Bacillus sp, Klebsiella sp, V.
harveyi, V. parahaemolyticus).
• The biosurfactant was characterized by TLC, HPLC &
NMR.
• Further studies on biosurfactants, provides tremendous
applications in industrial fields as detergents in
biomedical and therapeutical fields.
ACKNOWLEDGEMENT
•
I owe to almighty God and my beloved mother for their blessings and help, without which I might never
have been successful.
•
It is my privilege to express my deep sense of gratitude and sincere thank to my guide Dr. S. Satheesh,
Lecturer, International Centre for Nanobiotechnology, Rajakkamangalam for providing necessary facilities
to carry out my project in this institution.
•
It is my privilege to express my deep sense of gratitude and sincere thank to my co-guide Dr. S. G.
Prakash Vincent, Associate Professor, for his invaluable contribution, encouragement, optimistic
approach in any problem and systematic guidance to make this out a successful project.
•
I take this opportunity to express my hearty thanks to Dr.L.Uma, Head Department of Marine
Biotechnology, Bharathidasan University, Trichy, Dr.S.Mary Josephine Punitha, Assistant Professor,
Dr.G. Immanuel, Assistant Professor, Dr. T. Citarasu, Assistant Professor for their encouragement and
support throughout this study.
•
I would like to express my hearty thanks to my beloved friends and scholars who helped me directly and
indirectly for the completion of my project work.
•
I wish to express my sense of appreciation to my sisters who patiently endured and gave encouragement
and support throughout my studies.
•
Once again I thank to god for making everything possible with the help of our guides, family members,
friends and everyone who helped me. (K.Pandi selvi)
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