SSStttooonnnyyy BBBrrrooooookkk UUUnnniiivvveeerrrsssiiitttyyy The official electronic file of this thesis or dissertation is maintained by the University Libraries on behalf of The Graduate School at Stony Brook University. ©©© AAAllllll RRRiiiggghhhtttsss RRReeessseeerrrvvveeeddd bbbyyy AAAuuuttthhhooorrr... The Impact of Environmental Factors on the Physiology of the Atlantic surfclam, Spisula solidissima A Thesis Presented By Jesse Hornstein to The Graduate School in Partial Fulfillment of the Requirements for the Degree of Master of Science in Marine and Atmospheric Science Stony Brook University August 2010 Stony Brook University The Graduate School Jesse Hornstein We, the thesis committee for the above candidate for the Master of Science degree, hereby recommend acceptance of this thesis. Dr. Bassem Allam Thesis Advisor Associate Professor School of Marine and Atmospheric Sciences Dr. Emmanuelle Pales Espinosa Adjunct Assistant Professor School of Marine and Atmospheric Sciences Dr. Robert M. Cerrato Associate Professor School of Marine and Atmospheric Sciences Dr. Kamazima M.M. Lwiza Associate Professor School of Marine and Atmospheric Sciences This thesis is accepted by the Graduate School Lawrence Martin Dean of the Graduate School ii Abstract of the Thesis The Impact of Environmental Factors on the Physiology of the Atlantic surfclam, Spisula solidissima By Jesse Hornstein Master of Science in Marine and Atmospheric Science Stony Brook University 2010 The Atlantic surfclam supports a multi-million dollar fishery in New York. Between 2002 and 2008, surfclam abundance in New York State waters along the south shore of Long Island has decreased by 63% in biomass and 75% in the total number of clams. Population age structure has drastically shifted since 2002 indicating the lack of recruitment. Only 3% of the population was made up of clams that were 6 years old and younger in 2006, compared to 21% in 2002. In 2008, 15% of the population was composed of clams less than 6 years in age. It is hypothesized that increased temperatures in recent years has caused stress in these animals, negatively impacting their physiology leading to a reduction in population size. Studies were conducted on surfclam energy balance, scope for growth and immune function to investigate the impact of temperature on the physiology of these iii animals. Results suggest energy reserves are used differently during warm and cooler years, which may impact survival and reproductive success. Further studies on scope for growth indicate an increased metabolic demand at 23°C compared to 19°C. Results also demonstrated a reduction in filtration rate at 23°C compared to 19°C which could cause an energetic imbalance during the critical period following spawning. Furthermore, short term energy usage was greater at 23°C, and data from immune defense studies imply surfclams are immuno-compromised at this temperature. These results strongly suggest that stressful summer temperatures negatively influence surfclam physiology. iv Table of Contents List of Figures viii List of Tables xi Introduction 1 Objectives 8 Methods 10 Examination of historical population and temperature data 10 Health surveillance of the current population 10 The impact of temperature on gonadal development 11 Filtration rate and scope for growth 13 Immune defense and short term energy balance 14 Histology/Condition Index 15 Biochemical analyses: 15 Glycogen 16 Lipids 17 Protein 18 Vitellogenin 18 Scope for Growth Analyses: 19 Filtration Rate 19 Ingestion Rate 20 Assimilation Efficiency 20 Oxygen Consumption 20 Ammonia Excretion 21 Irrigatory Efficiency 21 Scope for Growth 22 Immunological Analyses: 22 Bacteriology 22 Differential Hemocyte Counts 23 Phagocytosis 23 Reactive Oxygen Species (ROS) 24 Viability 24 Statistical Methods 25 Results 26 Examination of historical population and temperature data 26 Health surveillance of the current population: 26 Histology/Condition index 26 Gonad Biochemistry (2008) 28 Monthly Variability 28 Males/Females 28 Normal/Abnormal 29 Adductor Muscle Biochemistry (2009) 30 Monthly Variability 30 v Males/Females 30 Normal/Abnormal 31 Gill Biochemistry (2009) 31 Monthly Variability 31 Males/Females 31 Normal/Abnormal 31 Gonad Biochemistry (2009) 32 Monthly Variability 32 Males/Females 32 Normal/Abnormal 33 Mantle Biochemistry (2009) 34 Monthly Variability 34 Males/Females 34 Normal/Abnormal 35 Gonad Biochemistry 2008 vs. 2009 35 Immune parameters 35 Bacteriology 36 The impact of temperature on gonadal development: 36 Histology/Condition index 36 Biochemistry 36 Filtration rate and scope for growth 37 Immune defense and short term energy balance: 38 Biochemistry/Condition Index 38 Immune Parameters 39 Discussion 41 Examination of historical population and temperature data 41 Health surveillance of the current population: 44 Histology/Condition index 44 Gonad Biochemistry (2008) 46 Monthly Variability 46 Males/Females 47 Normal/Abnormal 48 Adductor Muscle Biochemistry (2009) 50 Monthly Variability 50 Gill Biochemistry (2009) 50 Monthly Variability 50 Gonad Biochemistry (2009) 51 Monthly Variability 51 Males/Females 52 2008 vs. 2009 52 Mantle Biochemistry 54 Monthly Variability 54 Biochemistry of normal and abnormal individuals 54 Summary 55 vi Immune Parameters 56 Bacteriology 57 The impact of temperature on gonadal development: 57 Histology/Condition index 57 Biochemistry 58 Filtration rate and scope for growth 59 Immune defense and short term energy balance: 61 Biochemistry 61 Immune parameters 62 Future Directions 64 Tables 68 Figures 71 References 101 Appendix A: A summary of gonad conditions detected in processed animals 111 Appendix B: Semi-quantitative used for the evaluation of abnormal gonadal development (Kim and Powell, 2004) 111 Appendix C: Condition index of normal and abnormal clams in 2008 and 2009 112 Appendix D: Glycogen, Total Carbohydrate, Lipid and Protein levels of male and female surfclam adductor muscles in 2009 112 Appendix E: Glycogen, Total Carbohydrate, Lipid and Protein levels of normal and abnormal surfclam adductor muscles in 2009 113 Appendix F: Glycogen, Total Carbohydrate, Lipid and Protein levels of male and female surfclam gills in 2009 113 Appendix G: Glycogen, Total Carbohydrate, Lipid and Protein levels of normal and abnormal surfclam gills in 2009 114 Appendix H: Glycogen, Total Carbohydrate, Lipid and Protein levels of male and female surfclam mantles in 2009 114 Appendix I: Glycogen, Total Carbohydrate, Lipid and Protein levels of normal and abnormal surfclam mantles in 2009 115 Appendix J: Histological data from the impact of temperature on gonadal development study 115 Appendix K: Statistical tests and results 116 Appendix L: Histological rankings of male gonads 124 Appendix M: Histological rankings of female gonads 125 Appendix N: Histological snapshots of abnormal gonad conditions, parasites and hemic neoplasia 126 vii List of Figures Figure 1: Percent seed (clams <100 mm length) in NYS DEC surfclam population assessments from 1992-2008 71 Figure 2: Average summer temperatures between 2000 and 2009 71 Figure 3: Gametogenic stages of clams collected in 2008 72 Figure 4: Gametogenic stages of clams collected in 2009 72 Figure 5: Condition index and sea surface temperature in 2008 (A), 2009 (B) and a comparison of condition index in 2008 vs. 2009 (C) 73 Figure 6: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of surfclam gonads in 2008. 74 Figure 7: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of male and female surfclam gonads in 2008 75 Figure 8: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of normal and abnormal surfclam gonads in 2008 76 Figure 9: Glycogen (A), Total Carbohydrate (B), Lipid (C) and Protein (D) levels of surfclam adductor muscles in 2009 77 Figure 10: Glycogen (A), Total Carbohydrate (B), Lipid (C) and Protein (D) levels of surfclam gills in 2009 78 Figure 11: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of surfclam gonads in 2009 79 Figure 12: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of male and female surfclam gonads in 2009 80 Figure 13: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of normal and abnormal surfclam gonads in 2009 81 Figure 14: Glycogen (A), Total Carbohydrate (B), Lipid (C) and Protein (D) levels of the surfclam mantle in 2009 82 Figure 15: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of surfclam gonads in 2008 and 2009 83 Figure 16: Differential hemocyte counts (A) and the percent granulocytes (B) from field samples in 2009 84 viii Figure 17: Bacteria counts (per 100µl hemolymph) from the field on Maine agar (A.), TCBS (B.), and CHROMagar (C.) medias 85 Figure 18: Gametogenic stages of clams from the impact of temperature on gonadal development study 86 Figure 19: Condition index of clams in the cold and warm treatments from the impact of temperature on gonadal development study 86 Figure 20: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of surfclam gonads from the impact of temperature on gonadal development experiment 87 Figure 21: Glycogen (A), Total Carbohydrate (B), Lipid (C), Protein (D) and Vitellogenin (E) levels of male and female surfclam gonads from the impact of temperature on gonadal growth experiment. 88 Figure 22: