University of Massachusetts Amherst ScholarWorks@UMass Amherst Masters Theses 1911 - February 2014 1989 Assimilation of different foods by larvae of Simulium verecundum Stone and Jamnback (Diptera: Simuliidae). Paula J. S. Martin University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/theses Martin, Paula J. S., "Assimilation of different foods by larvae of Simulium verecundum Stone and Jamnback (Diptera: Simuliidae)." (1989). Masters Theses 1911 - February 2014. 3057. Retrieved from https://scholarworks.umass.edu/theses/3057 This thesis is brought to you for free and open access by ScholarWorks@UMass Amherst. It has been accepted for inclusion in Masters Theses 1911 - February 2014 by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. UMASS/AMHERST o 3130bb007b4E!lfl3 ASSIMILATION OF DIFFERENT FOODS BY LARVAE OF SIMULIUM VERECUNDUM STONE AND JAMNBACK (DIPTERA : SIMULIIDAE) A Thesis Presented by PAULA J.S. MARTIN Submitted to the Graduate School of the University of Massachusetts in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE February 1989 Department of Entomology Copyright by Paula J.S. Martin 1989 All Rights Reserved ASSIMILATION OF DIFFERENT FOODS BY LARVAE OF SIMULIUM VERECUNDUM STONE AND JAMNBACK (DIPTERA : SIMULIIDAE) A Thesis Presented by PAULA J.S. MARTIN Approved as to style and content by: Dedicated to Joe For pursuing the red scarf on the hill and making me forever happy ACKNOWLEDGEMENTS There are a number of people who have helped in the development and completion of this thesis. First and foremost is Dr. John D. Edman, my major advisor. Without John s guidance, patience and assistance, especially through the numerous moments of frustration, this thesis would never been completed. The freedom of choice for research topic was a bounty I will always appreciate. Thank you, John. My graditude also goes to the other members of my committee. Drs. John M. Clark and Dave E. Leonard. Both have provided me with valuable advise on this research. My appreciation goes to my other advisees including Drs. Rich W. Merritt, Roger Wotton, Dan Molloy, Doug Craig and Ned Walker . Special thanks must go to John Clark and the people in his lab: Jacques Marion, Joe Argentine and Feng Guo-Lei. Along the way, they shared space, equipment, double- distilled water and a few laughs. I wish to especially acknowledge the Apiary Gang. The members of this illustrious group, past and present, include: Russell Coleman, Sangvorn Kittawee, Ken Simmons, Sue Opp, Pat Kittayapong, Rob Anderson and Dennis LaPointe. All have added something to my life at UMass. Russ entertained us all with his concocted stories we never v believed (well, hardly ever). Sang always provided a ready laugh and smile. Dennis not only assisted with this research but has been a steady friend. Because of Dennis I will never again peruse the newspaper in quite the same way. I must especially thank my family for their moral support in this venture. They kept to a minimum the question, "So, when are you going to be done?" Particular thanks goes to my mother for keeping me going when my enthusiasm lagged. Lastly, I must thank my husband Joe for his absolute support of my work and performance. Great comfort and strength has been derived from his unqualified love and aid. vi ABSTRACT ASSIMILATION OF DIFFERENT FOODS BY LARVAE OF SIMULIUM VERECUNDUM STONE AND JAMNBACK (DIPTERA : SIMULIIDAE) February 1989 PAULA J.S. MARTIN B•S./ University of California, Berkeley M.S., University of Massachusetts, Amherst Assimilation of different food materials by a lotic filter feeder, Simuliurn verecundum, was studied. Ingestion and assimilation of dissolved organic matter (DOM) was determined with radiolabelled glucose. Extremely low radiolabel uptake led to the inclusion of Aedes aegypti. a filter feeder that is known to drink, as an experimental control. Radiolabel uptake by S_._ verecundum did not change for larvae fed in absense of current or with plugged mouthparts versus normal larvae. Radiolabel uptake by Aedes «? 1 "*. "" larvae with plugged mouthparts was < 3% of radiolabel uptake in normal larvae. Aedes aegypti was able to ingest and assimilate the DOM. Simulium verecundum, Erosimuliurn mixturn/fuscum and Cnephia dacotensis were unable to ingest DOM. The rationale for this difference is in the vi i different filter-feeding mechanism of Culicidae and Simuliidae. Culicidae use their filtering apparatus, i.e. mouth brushes, to create a current and to filter particulates; therefore they ingest water with particulate food. Simuliidae use their filtering apparatus, i.e. cephalic fans, only to filter food; a solid food bolus is ingested. Assimilation of different particulate organic matter (POM) was tested with radiolabelled algae, Navicula and Scenedesmus, and size-fractionated bacterioplankton. All were ingested and assimilated. Navicula and bacterioplankton had assimilation ratios, a measure of assimilation rate, 3 times that of the green alga, Scenedesmus. Cell wall composition and black fly gut pH may be the reason for these differences. vi 11 TABLE OF CONTENTS Page ACKNOWLEDGEMENTS. v ABSTRACT. vi i LIST OF TABLES. » . xi LIST OF FIGURES . xi i Chapter I. INTRODUCTION . A. Preface. H •••••••••• B. Literature Review. (N 1. Filter Feeding.. (N a. How do filter-feeders function within CN a stream ecosystem? . b. How do filterers change particle size? . ro fO (1) Filtering mechanism . i£) (2) Filtering effects on POM. c. How do black flies fit into the filter-feeding picture? . 11 (1) Filtering mechanism . 11 (2) Habitat and food resources of black flies.13 2. Assimilation.. a. Terminology.17 b. Methodology.18 c. Black fly assimilation.21 3. Seston Composition . 22 II. DISSOLVED ORGANIC MATTER INGESTION BY FILTER-FEEDING DIPTERA.24 A. Introduction.24 B. Materials and Methods. 26 1. Experimental Design.26 2. Test Animals.27 3. Radiolabel Determination . 29 C. Results.38 D. Discussion.42 ix III. PARTICULATE ORGANIC MATTER ASSIMILATION BY SIMULIUM VERECUNDUM . 54 A. Introduction.54 B. Materials and Methods.55 1. Experimental Design.55 2. Test Animals.. 3. LSC Procedure.53 4. Foods Tested.o'4 a. Bacteria.. b. Algae.76 5. DAPI staining procedure.80 6. Protein Analysis . 84 C. Results.85 D. Discussion.101 IV. CONCLUSIONS.106 APPENDIX: COMPOSITION OF WOODS HOLE MBL MEDIA.108 BIBLIOGRAPHY.. \ x LIST OF TABLES Page Table 1. Radiolabel Counts Associated with Black Flies, 6th-7th Instar Larvae, Fed C-Labelled, Dissolved Glucose . 39 Table 2. Effect of Larval Instar on Radiolabel Counts Associated with S_. verecundum Fed C—Labelled Dissolved Glucose.39 Table 3. Radiolabel Counts Associated with 3e_gypt]^, 4th Instar, 7 Day Old Larvae, Fed C-Labelled, Dissolved Glucose . 40 Table 4. Radiolabel Counts Associated with Larvae Fed C—Labelled, Dissolved Glucose under Experimental Conditions to Block or Stop Feeding, 3 hr Feeding Period.41 Table 5. Radiolabel Counts Associated with Larvae Fed 14C-Labelled, Dissolved Glucose with Inert Particulates Present to Induce Feeding, 3 hr Feeding Period. Table 6. Assimilation Ratios. Radiolabel Incorporated into Tissue in 5 hr Feeding Period per Gut Contents, for S_^_ ver ecundum Fed Radiolabelled, Particulate Foods.85 Table 7. Assimilation Efficiencies. Radiolabel Incorporated into Tissue per Total Radiolabel Ingested in 5 hr Feeding Period for S. verecundum Fed Radiolabelled, Particulate FOOdS. > or Table 8. Cell and Radiolabel Concentration of Radiolabelled Particulate Food Fed to S. yer_ecundu,m in Different Experiments.87 Table 9. Cells and Carbon Incorporated into Tissue and Assimilation Rate and Gut Contents of S. verecundum Larvae Fed Radiolabelled Navicula over Time.. Table 10. Number of Cells Assimilated in a 5 hr Feeding Period by S_,__ verecundum Larvae Fed Radiolabe 1 led, Particulate Foods.89 Table 11. Protein Content of Cultured Algae and Field-Collected Bacteria. 90 LIST OF FIGURES Page Figure 1. Quench Curve for Aqueous Sample with Low l^C-Glucose Concentration (< 10000 dpm/vial). 31 Figure 2. Quench Curve for Aqueous Sample with High ^C-Glucose Concentration (> 25000 dpm/vial). 33 Figure 3. - Quench Curve for NCS Solubilizer with Digested Larvae and High ^C-Glucose Concentration (> 25000 dpm/vial).35 Figure 4. DOM-Associated Label Counts versus Wet Weight of S_._ verecundum after 1 and 3 hr Feeding Periods . 43 Figure 5. DOM-Associated Label Counts versus Time for A . aegypt i.4 5 Figure 6. Comparison of DOM-Associated Label Counts versus Time for verecundum and A_._. aegypti, + 1 S.E..-.47 Figure 7. Comparison of DOM-Associated Label Counts, Logarithmic Scale, versus Time for S. verecundum and A_^_ aegypti, + 1 S.E.49 Figure 8. Feeding Rate Variation. Frequency Distribution of Bacterial Cells per Gut Content.57 Figure 9. Feeding Rate Variation. Frequency Distribution of Navicula Cells per Gut Content.. • Figure 10. Feeding Rate Variation. Frequency Distribution of Scenedesmus Cells per Gut Content . Figure 11. Quench Curve for Aqueous Sample with Algae and Low Na214C03 Concentration (< 2200 dpm/vial) . Figure 12. Quench Curve for Aqueous Sample with Algae and High Na214CC>3 Concentration (> 22000 dpm/vial) . XI 1 Figure 13. Quench Curve for Aqueous Sample with Bacteria and methyl-[]-thymidine Concentration > 4000 dpm/vial . 69 Figure 14. Quench Curve for NCS Solubilizer with Digested Larvae and Na2^C03 Concentration < 2000 dpm/vial.VI Figure 15. Quench Curve for NCS Solubilizer with Digested Larvae and methyl-[]-thymidine Concentration > 6000 dpm/vial.73 Figure 16. Flowchart for Radiolabelling Bacter i oplankton.77 Figure 17. Flowchart for Radiolabelling Algae .... 81 Figure 18. Navicula Cells Assimilated versus Time . 91 Figure 19* Navicula Cells in Gut Content versus Time. 93 Figure 20. Radiolabel Respired for Navicula and Scenedesmus.95 Figure 21. Navicula Radiolabel Respired versus Time . 97 Figure 22. Protein Analysis Standard Curve.99 xi i i CHAPTER I INTRODUCTION A. Preface Filter-feeding aquatic black fly (Simuliidae) larvae play an important role in energy flow in stream ecosystems.