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The Effect of Arsenic Trioxide on the Grey Flesh Fly Sarcophaga Bullata (Diptera: Sarcophagidae)

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The Effect of Arsenic Trioxide on the Grey Flesh Fly Sarcophaga Bullata (Diptera: Sarcophagidae) The Effect of Arsenic Trioxide on the Grey Flesh Fly Sarcophaga bullata (Diptera: Sarcophagidae) by Nina Dacko B.S. A Thesis In ENVIRONMENTAL TOXICOLOGY Submitted to the Graduate Faculty of Texas Tech University in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Approved Steven M. Presley PhD Committee Chair Stephen B. Cox PhD George P. Cobb PhD Peggy Miller Dean of the Graduate School May, 2011 Copyright 2011, Nina Dacko Texas Tech University, Nina Dacko, May 2011 ACKNOWLEDGMENTS First and foremost, I would like to thank my advisor, Dr. Steven Presley, for he has been utterly helpful in thesis guidance and funding as well as with side work and professional connections. May I one day, follow in your footsteps as a medical entomologist. I look up to you as a scientist, a friend, and as a father figure. I would like to thank committee member, Dr. George Cobb who gave me more than adequate advice about chemical analysis as well as suggestions for statistical analysis and suggested contacts for advice and knowledge pertaining to my research. I must admit being intimidated by your intelligence, but you were always easy to understand. Also thank you for your numerous suggestions in thesis writing. I would like to thank committee member Dr. Stephen Cox who suggested statistical analyses, explained why these analyses correspond to presented research questions and most of all helped me to recognize why these analyses were superior to others in similar research. I also give credit to Stephen for helping me in logical thesis and defense organization skills. I would like to thank the entire vector-borne zoonoses lab (VZL): specifically Anna Hoffarth, Juliet Kinyua, James Walls and Anna Gibson for their help, watching my fly colonies while I was away and Anna Gibson for teaching me numerous tasks in the laboratory- dissection of rabbits and oral dosing- for my thesis along with PCR, RNA isolation, colorimetric protein assays and other things for work experience. Others at TIEHH include Richie Ericson- for statistics suggestions, and Dawn Slekis- again for fly care. Thanks to Ben Wozniak and Applied Speciation in Spokane, WA for performing arsenic speciation analyses as well as giving me suggestions for my own chemical ii Texas Tech University, Nina Dacko, May 2011 analyses. You’ve been most helpful and a lot of results were dependent on the information you have provided me at no charge! Dr. Galen Austin- thank you for insectary maintenance, such as fixing doors and setting up temperature monitors and for lending me your digital caliper. I would also like to thank Mitchell Burt for helping me build fly cages and constantly helping with heavy carboys. Thanks to Amber Matthews and Mark Goza for rabbit housing and maintenance and euthanasia facilities. Thanks to Dr. Marjolaine Giroux for identification of Sarcophaga bullata, to Dr. Lois Held for supplying fly standards in chemical analysis and James Cockendolpher for fly identification knowledge. Thanks to professors and colleagues who frequently answered questions when asked including- Dr. Greg Mayer, Dr. Todd Anderson and Dr. Emily Notch. Thanks to Dr. Jaclyn Cañas for use of the metal analysis laboratory and Brad Thornhill and Marshall Pattee for help with the ICP-MS. Thanks to Shibin Li and Shuanying Yu for help and advice with digestions. Thanks to my professors in the plant and soil science department and at the forensic science institute. Thanks to Michael Lee Goff for experimental suggestions and inspiration. Thanks to Dr. RJ Kendall and TIEHH for facilities. Thanks Faculty and Staff- Matthew Young for being curious! iii Texas Tech University, Nina Dacko, May 2011 Thanks friends and family and Rob Bendkowski for driving me to the insectary in snow storms and sitting in the stinky insectary while I worked. iv Texas Tech University, Nina Dacko, May 2011 TABLE OF CONTENTS ACKNOWLEDGMENTS ............................................................................................. ii ABSTRACT ............................................................................................................. vii LIST OF TABLES ................................................................................................... viii LIST OF FIGURES ................................................................................................... ix I. INTRODUCION ...................................................................................................... 1 Arsenic History in Industry, Crime, Warfare, Environment and Medicine ........ 1 Postmortem Interval, Forensic Entomology and Entomotoxicology .................. 3 Life History of Sarcophaga bullata .................................................................... 5 Past Research in the Field of Entomotoxicology ................................................ 7 Case Studies ............................................................................................................................ 7 Laboratory Studies ................................................................................................................ 10 Gaps in Research ............................................................................................... 16 Objectives, Aims and Hypotheses .................................................................... 17 Literature Cited ................................................................................................. 18 II. MATERIALS AND METHODS ............................................................................ 23 Rabbits .............................................................................................................. 23 Chronic Exposure .................................................................................................................. 23 Acute Exposure ..................................................................................................................... 24 Flies ................................................................................................................... 24 Collection .............................................................................................................................. 24 Colony Establishment ........................................................................................................... 25 Larval exposure and measurements ...................................................................................... 25 Pupae and adult maintenance, reproductive output and generation two ............................... 25 Chemical Analyses ............................................................................................ 26 Arsenic speciation and analyses ............................................................................................ 26 Digestion ............................................................................................................................... 26 Total arsenic analyses ........................................................................................................... 27 Statistycal analyses ........................................................................................... 27 Fly growth rate and development trends ............................................................................... 27 Fly life paramters and arsenic concentrations in tissues........................................................ 28 Literature Cited ................................................................................................. 28 III. RESULTS AND DISCUSSION ............................................................................. 29 Observations ...................................................................................................... 29 Fly growth rate and development trends ........................................................... 31 Emergence and metamorphic trends ................................................................. 43 Mortality ............................................................................................................ 45 Arsenic in tissues .............................................................................................. 47 v Texas Tech University, Nina Dacko, May 2011 Reproductive Output ......................................................................................... 53 Literature Cited ................................................................................................. 54 III. CONCLUSIONS ................................................................................................ 55 Literature Cited ..................................................................................................56 A. ARSENIC MSDS AND DIGESTION METHOD........................................................ 57 B. RAW DATA ........................................................................................................ 74 vi Texas Tech University, Nina Dacko, May 2011 ABSTRACT Larvae of Sarcophaga bullata (Diptera: Sarcophagidae), a necrophagous insect commonly utilized in the field of entomotoxicology, were reared on rabbit tissues of rabbits that were previously exposed to arsenic trioxide (As2O3) by different methods. We observed the effects of arsenic (As) and As metabolites in acute versus chronic exposure in rabbit tissues on growth rate (mean larval length), metamorphosis, mortality and reproductive output of S. bullata. The New Zealand white rabbit was utilized as a vehicle to create natural concentrations
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