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Title
CERTIFICATE
DECLARATION
ACKNOWLEDGEMENT
CONTENTS
ABBREVIATIONS
Preface
Introduction
1.1 Biomarkers
1 2 Stress Proteins as Biomarkers
1.3 Free Radicals and Protein Damage
1.4 References
Materials and Methods
2.1 Protein Estimation
2.1.1Bradford Assay
2.1.2 Lowry Method
2.2 Determination of Stress Proteins
2 2 1 Poly Acrylamide Gel Electrophoresis
2 2.2 Sample Preparation
2.2.3. Performance of Gel Electrophoresis
2.2.4 Staining of Gel for Proteins
2 3 Western Blot Analysis
2.3.1 Electrophorectic Transfer of Proteins on to Nitrocellulose Membrane
2.3.2 Blot Development
2.4 Enzyme Linked lmmunosorbant Assay (ELISA) for Quantitative Determination of HspGO Induction
2 5 Effects of Free Radicals
2 5 1 Estimation of Thiobarbituric Acids Reacting Substances (TBARS)
2 5 2 Determination of Carbonyl Content in Oxidatively Modified Proteins
2 6 Antioxidant Enzyme Systems
2 6 1 Estimation of Superoxide Dismutase
2 6 2 Estimation of Catalase
2 6 3 Estimation of Glutathione Peroxidase (GSH-Px)
2 6 4 GSH Estimation
2 6 5 Estimation of Glutathione-S-transferase (GST)
2.7 Isozyme Analysis
2 7 1 Super Oxide Dismutase Isozyme Analysis
2 7 2 Catalase Isozyme Analysis
2 8 The Stressors Used
2 9 Statistical Analysis
2 10 References
Stress Responses in Cyanobacteria- Spirulina platensis
3.1 Introduction
3.1 1 Heat Stress
3.1 2 Salinity Stress
3.1.2.1 Salinity and Osrnoprotectants
3.1.2.2 Effects on lntracellular Ionic: Balance
3.1.2.3 Effects on Photosynthesis
3.1.2.4 Salt Induced Changes in Gene Expression
3 1 3 Effects of Pesticides on Cyanobacteria
3.1.3.1. Effect on Survival 8 Growth
3.1.3.2 Effect on Metabolic Processes
3.1.3.2.1 Photosynthesis and Respiration
3.1.3.2.2 Nitrogen Fixing Ability
3 1 4 Use of Cyanobacteria in Pollution Abatement
.3.2 Materials and Methods
3 2 1 Maintenance Of Spirulina Platensis
3 2.2 Stress Induced Growth Inhibition Studies,
3.2.2.1 Chlorophyll Estimation
3 2 3 Sample Preparation
3.2.3.1 Determination of Stress Proteins
3.2.3.2 Quantification of Hsp 60 Expression
3 2.4 lsoenzyme Analysis
3 2 5 Effects of Free Radicals
3.2.5.1 Protein Carbonyl Estimation
3.2.5.2 TBARS Estimation
3.2.6 Antioxidant Systems
3.2.6.1 SOD Estimation
3.2.6.2 Catalase Estimation
3.2.6.3 Glutathione (GSH) Content
3.2.6.4 Peroxidase Activity
3.2.6.5 GST Estimation
3 3 Results
3.3.1 Stress Induced Growth Inhibition Studies
3.3.2 Stress Protein Induction in Spirulina platensis Under Various Stressors
3.3.2.1 SDS PAGE and Western Blot Analysk
3.3.2.2 Dose and Time Dependent Expression of Hsp6O
3.3.3 Stressor Induced Potential for Oxidative Stress
3.3.3.1 Temperature Stress
3.3.3.2 Salinity Stress
3.3.3.3 Paraquat Induced Stress
3.3.3.4 Endosulfan Induced Stress
3.3.3.5 Malathion Induced Stress
3.3.3.6 Dimethoate Induced Stress
3 3 4 Antioxidant lsoenzymes Analysis
3.4 Discussion
3 4 1 Growth Inhibition Studies
3 4 2 Stress Protein Induction Studies
3 4.3 Influence of Oxidative Stress in Stress Protein Induction in Spirulina platensis
3 5 Summary
3 6 References
Stress Responses in Fresh Water Bivalve- Mollusc Lamellidens marginalis (Bloch.)
4.1 Introduction
4.1.1Lysosomal Alterations
4.1.2 Cell Type Replacement
4.1.3 Imposex
4. 1.4 Metallothionein Induction
4.1.5 Acetylcholinesterase
4.1. 6 Stress Proteins
4.2 Materials and Methods
4.2 1 Maintenance of Mussel Lamellidens marginalis (Bloch.)
4. 2. 2 96 hour LC50 Determination
4.2 3 Sample Preparation
4.2.3.1 Determination of Stress Proteins
4.2.3.2 Quantification of Hsp6O Expression
4.2.3.3 lsoenzyme Analysis
4.2.3.4 Effects of Free Radicals
4.2.3.4.1 Protein Carbonyl Estimation
4.2.3.4.2 TBARS Estimation
4.2.3.5 Antioxidant Systems
4.2.3.5.1 SOD Estimation
4.2.3.5.2 Catalase Estimation
4 3 Results
4.3.1 Profit Analysis of Toxicity Study on Lamellidens marginalis
4.3 2 Stress Protein Induction in Lamellidens marginalis Under Various Stressors
4.3.2.1 SDS PAGE and Western Blot Analvsis
4.3.2.2 Dose and Time Dependent Expression of Hsp6O
4 3 3 Stressor Induced Potential for Oxidative Stress
4.3.3.1 Temperature Induced Stress
4.3.3.2 Salinity Induced Stress
4.3.3.3 Paraquat Induced Stress
4.3.3.4 Endosulfan Induced Stress
4.3.3.5 Malathion Induced Stress
4.3.3.6 Dimethoate Induced Stress
4 3 4 Antioxidant Isoenzyme Analysis
4 4 Discussion
4 4 1 96 hour LC50 Toxicity Study on Lamellidens marginalis
4 4 2 Stress Protein Induction Studies
4.4.3 Influence of Oxidative Stress in Stress Protein Induction in Lamellidens marginalis
4.4.3.1 Temperature Induced Stress
4.4.3.2 Salinity Induced Stress
4.4.3.3 Paraquat Induced Stress
4.4.3.4 Endosulfan lnduced Stress
4.4.3.5 Malathion lnduced Stress
4.4.3.6 Dimethoate Induced Stress
4.5 Summary
4.6 References
Stress Responses in Fresh Water FishAnabas testudineus (Bloch.)
5.1 Introduction
5 1 2 Oxidative Metabolism
5.1.3Heat Shock Proteins
5.2 Materials and Methods
5.2.1 Maintenance of Fish Anabas testudineus (Bloch.)
5.2.2 96 hour LCw, Determination
5.2.3 Sample Preparation
5.2.3.1 Determination of Stress Proteins
5.2.3.2 Quantification of Hsp6O Expression
5.2.4 Effects of Free Radicals
5.2.4.1 Protein Carbonyl Estimation
5.2.4.2 TBARS Estimation
5.2.5 Antioxidant Systems
5.2.5.1 SOD Estimation
5.2.5.2 Catalase Estimation
5.2.5.3 Glutathione (GSH) Content:
5.2.5.4 Glutathione Peroxidase (GSH-Px) Activity
5.2.5.5 GST Estimation
5.3 Results
5.3.1 Probit Analysis of Toxicity Study of Fish
5 3 2 Stress Protein Induction in Anabas testudineus Under Various Stressors
5.3.2.1 SDS PAGE and Western Blot Analysis
5.3.2.2 Dose and Time Dependent Expression of Hsp6O
5.3.3 Stressor Induced Potential for Oxidative Stress
5.3.3.1 Temperature Induced Stress
5.3.3.2 Salinity Induced Stress
5.3.3.3 Paraquat Induced Stress
5.3.3.4 Endosulfan Induced Stress
5.3.3.5 Malathion induced Stress
5.4 Discussion
5.4.1 96 hour LC50 Toxicity Study on Anabas testudineus
5.4.2 Stress Protein Induction Studies
5 4 3 Influence of Oxidative Stress in Stress Protein Induction in Anabas testudineus
5.4.3.1 Temperature Induced Stress
5.4.3.2 Salinity Induced Stress
5.4.3.3 Paraquat Induced Stress
5.4.3.4 Endosulfan Induced Stress
5.4.3.5 Malathion Induced Stress
5.5 Summary
5 6 References