Chitobiase As a Surrogate Measure of Aquatic Invertebrate Biomass And

Chitobiase as a surrogate measure of aquatic invertebrate biomass and secondary production in an environmental effects monitoring context by Matthew Randell A Thesis Submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfillment of the requirements of the degree of MASTER OF SCIENCE Department of Environment and Geography University of Manitoba Winnipeg Copyright © 2019 by Matthew Randell ABSTRACT The current techniques used to assess aquatic invertebrate community status in the field are typically labour and time intensive, and therefore the development and implementation of new rapid and cost-effective methodologies is warranted. A proposed option is the enzymatic assay to detect and quantify the rate of production of the molting enzyme chitobiase, which can be used for determining impacts on freshwater aquatic systems. Two case studies were performed at: 1) The Prairie Wetland Research Facility at the University of Manitoba, to determine if a relation exists between measures of chitobiase and aquatic invertebrate biomass in a mesocosm setting, as well as to determine if changes in chitobiase activity could detect impacts to aquatic invertebrate communities from sulfamethoxazole and diluted bitumen and; 2) in the Elk River Valley region of British Columbia, to determine if a positive relationship exists between the rate of chitobiase production and benthic invertebrate biomass in lotic freshwater systems. No significant relationship was observed between the chitobiase measures and invertebrate biomass measures, and no effects of the stressors were detected in the first study. A significant positive relationship was observed between the rate of chitobiase production and benthic invertebrate biomass in the second study. It is recommended that additional studies be performed to further assess the potential of chitobiase activity to be used in an environmental monitoring context. ii ACKNOWLEDGMENTS I would like to thank my advisor, Dr. Mark Hanson for his endless support, guidance, and knowledge throughout my time at the University of Manitoba and for always making time for his students. My committee members Dr. CJ Mundy and Dr. Gordon Goldsborough for their guidance and patience. The Ecological Stress Laboratory members Adam Vanderpont, Angela Reeves, Chelsea Lobson, Dana Moore, and Jennifer Leslie. None of this would have been possible without their help. The team at Minnow Environmental Inc.: Dr. Michael White, Dr. Shari Weech, Kevin Martins, Tyrell Worrall, Jess Tester, Katharina Batchelar, Lisa Brown, Justin Wilson, and Dr. Heidi Currier for all their help and making me feel like part of the team during the field work in Elk Valley, British Columbia. Dr. Cheryl Podemski and Jian Zhang at DFO for their help and for allowing me to use their facilities. The environmental group at Tetra Tech Canada Inc.: Ryan Wizbicki, Brent Horning, Lena Andersson, Andrew Eason, Michel Grègoire, Jennifer Pieniuta, Ciara Sebelius, Kelly Jones, Ciro Longobardi, Lindsay Lutes, and Charlie Wu for their support and picking up the slack at work when I took time off to work on my thesis. My friends and family members for their encouragement, support, and curiosity throughout my studies. My parents, Dean and Connie Randell, for their support and encouragement. Finally, I would like to thank my wife, Amanda Randell, for her endless support, encouragement, and patience throughout my graduate studies program. iii CONTRIBUTIONS OF AUTHORS Chapter 3 is based on work conducted in the Elk Valley Region of British Columbia, Canada. Contributing authors include Matthew E. Randell1, who was involved in conducting the field study, analyzing the chitobiase samples, plotting and interpreting data, and writing the chapter. Michael S. White2, who was involved in conducting the field study, assisting with the statistical analysis of data collected, assisting in writing, and reviewing the chapter. Shari Weech2, who was involved in conducting the field study and reviewing the chapter. Patti Orr2, Carla Fraser3, and Mark L. Hanson1, who reviewed the chapter. iv TABLE OF CONTENTS ABSTRACT ........................................................................................................... ii ACKNOWLEDGMENTS ...................................................................................... iii LIST OF TABLES ................................................................................................. ix LIST OF FIGURES ............................................................................................. xiii CHAPTER 1. GENERAL INTRODUCTION .......................................................... 1 1.1 Environmental Effects Monitoring ........................................................... 3 1.1.1 Environmental Effects Monitoring Structure .......................................... 5 1.1.2 Biological Monitoring Studies ................................................................ 6 1.2 Benthic Invertebrates and their Use in Environmental Effects Monitoring ........................................................................................................ 9 1.2.1 Benthic Invertebrates ............................................................................ 9 1.2.2 Benthic Invertebrate Communities in Environmental Effects Monitoring ..................................................................................................................... 15 1.2.3 Disadvantages of the Benthic Invertebrate Community Survey .......... 19 1.3 Application of Chitobiase in Environment Effects Monitoring ............ 22 1.4 Chitin and Chitinolytic Enzymes ............................................................ 27 1.4.1 Sources of Chitobiase and Chitinolytic Enzymes in Aquatic Systems 28 1.4.2 Invertebrates ....................................................................................... 29 1.4.3 Bacteria ............................................................................................... 30 v 1.4.4 Fish ..................................................................................................... 30 1.4.5 Diatoms, Fungi and Protists ................................................................ 31 1.5 Factors Influencing Chitobiase in the Environment ............................. 32 1.5.1 Temperature ....................................................................................... 33 1.5.2 pH ....................................................................................................... 34 1.5.3 Discharge and Current ........................................................................ 34 1.5.4 Chemicals ........................................................................................... 35 1.6 Mesocosm Studies .................................................................................. 36 1.7 Compounds of Interest ........................................................................... 37 1.7.1 Sulfamethoxazole ............................................................................... 37 1.7.2 Diluted Bitumen .................................................................................. 39 1.8 Hypothesis ............................................................................................... 43 1.9 Research Objectives ............................................................................... 43 1.10 Literature Cited ...................................................................................... 45 CHAPTER 2. THE USE OF CHITOBIASE ACTIVITY AS A MEASURE OF AQAUTIC INVERTEBRATE BIOMASS: A FIELD-BASED MESOCOSM STUDY ............................................................................................................................ 61 2.1 Abstract .................................................................................................... 61 2.2 Introduction ............................................................................................. 62 2.3 Methods and Materials ............................................................................ 70 vi 2.3.1 Test facility .......................................................................................... 70 2.3.2 Sulfamethoxazole (‘SMX’) treatment and analyses ............................ 72 2.3.3 Diluted bitumen (‘dilbit’) treatment ...................................................... 73 2.3.4 Water quality parameters .................................................................... 73 2.3.5 Emergent insect sampling and analyses............................................. 74 2.3.6 Zooplankton sampling and analyses ................................................... 76 2.3.7 Chitobiase water sampling and analyses ............................................ 78 2.4 Statistical Analysis .................................................................................. 81 2.5 Results ..................................................................................................... 82 2.5.1 Sulfamethoxazole concentrations ....................................................... 82 2.5.2 Water quality parameters .................................................................... 83 2.5.3 Aquatic insect emergence enumeration and dry mass ....................... 84 2.5.4 Zooplankton biomass .......................................................................... 87 2.5.5 Total aquatic invertebrate biomass ..................................................... 88 2.5.6 Measured chitobiase activity ..............................................................

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