The Effect of Nicotine on Carrion Feeding Insects with Considerations For Use Within Forensic Sciences Andrew Chick A thesis submitted in partial fulfilment of the requirements of Nottingham Trent University, in collaboration with Staffordshire University, for the degree of Doctor of Philosophy. September 2014 - 1 - “Crime is common. Logic is rare. Therefore it is upon the logic rather than upon the crime that you should dwell” Sherlock Holmes. The Copper Beeches. By Sir Arthur Conan Doyle This work is the intellectual property of the author. You may copy up to 5% of this work for private study, or personal, non-commercial research. Any re-use of the information contained within this document should be fully referenced, quoting the author, title, university, degree level and pagination. Queries or requests for any other use, or if a more substantial copy is required, should be directed in the owner(s) of the Intellectual Property Rights. All Photos, unless expressly stated otherwise, are copy write of the author - 2 - Abstract The presence of invertebrates on decomposing animal matter has been used extensively by forensic entomologists to estimate time of death for over 100 years. The presence of toxins such as drugs and pesticides on carrion can affect the behaviour and life cycle of such invertebrates. The aim of this thesis was to examine the effects of nicotine upon the colonisation of carrion by invertebrates; nicotine was used because of its historical use as an insecticide and its ubiquitous use in society. The investigations aimed to examine these possible effects both in situ in field-based testing and ex situ in a controlled laboratory environment and to work towards an empirically testable correction factor for the estimation of Postmortem interval estimates in the presence of nicotine . The field-based testing was done using Sus domestica (Linnaeus) carrion with a solution of nicotine injected into the cadaveric throat of the animal. The carrion was protected from feeding and removal by vertebrate scavengers. It was found that nicotine affected the time taken for Diptera to colonise the carrion as well as affecting the behaviour of feeding. Diptera larvae showed avoidance of the nicotine treated throat sites on the carrion, which is the normal site of oviposition. It was determined that the rove beetle Creophilus maxillosus (Linnaeus) was exclusively found on the higher dose nicotine carrion. The rare hoverfly Rhingia rostrata (Linnaeus) was discovered on the control animal; this is the first specimen reported in Nottinghamshire. The investigation also found the first record of the Soldierfly Sargus bipunctatus breeding in carrion; the late breeding period of this species and its significance to the forensic entomologist is considered. The experiments were conducted in the Autumn/Winter months and Spring/Summer months. Nicotine appeared to have a differing effect with the season as the autumnal fauna varied from that of the spring fauna. The presence of nicotine appeared to prevent the animal carcass from drying out, typified by mycophagus beetles in autumn and semi-liquid habitat breeding flies in the summer. The laboratory based investigation examined the effects of nicotine upon the life cycle of Calliphora vomitoria including the effects upon oviposition, rate of development and survivablity. It was found that nicotine significantly affects rate of development of this forensically important fly. This study has shown that a careful study of a single chemical compound and its interaction with carrion and entomology has profound effects upon the alteration of the normal activity of a range of forensically important invertebrates. It will assist in improving the evidential usefulness of entomology to the Forensic Science and Policing communities in criminal investigations. - 3 - Acknowledgments NHL Legend Wayne Gretzky once said “you miss 100% of the shots you don’t take” but he also assisted twice as many goals as he scored. Without the following assistance I wouldn’t have had the chance to take this shot. Firstly I would like to thank my supervisory team, starting with the longest serving/suffering Director of Studies Dr Chris Terrell-Nield and the long suffering Prof John Cassella who have been part of the project really since before the start of the PhD, at the tail end of my MSc at Staffordshire University and in the case of Prof Cassella since the MSc, who have edited my numerous typos as well as edited and supported this work all the way through. Thank you to Prof Brian Pyatt, for his support and advice, for the first half of the project, as a Supervisor and the latter part as an advisor. Gratitude to Dr Chris Smillie and Dr Rachel Stubbington for their contributions after Brian’s retirement. Finally thanks to Dr David Hughes for his contribution as Independent assessor of the project. I am thankful to Nottingham Trent University for taking me on as PhD candidate, Staffordshire University for agreeing to be a collaborating institution, and The Universities of Derby and Lincoln for taking me on as an hourly paid lecturer which I feel has sharpened my skills in Forensic Entomology as well as my colleagues and friends at all of these institutions. I would like to thank the Dipterists’ Forum and Peter Chandler for allowing me to write the section on Collection of Diptera from Carrion for the Dipterists’ handbook, and the Royal Entomological Society for bestowing the honour of Fellowship upon me. But a PhD cannot thrive purely on the support and guidance of academics alone, and with this in mind I would like to thank my Family for their support and tolerance of their “weird relative that plays with bugs and dead things”, most notably my Mum and Dad (Chris and Sally Chick), My Gran (Beryl Chick), The outlaws (Pete and Maggie Sparks), Sam, Ash, Chelsea, Jessica, Brooke and Layla. You all helped keep me sane when I had difficult insects to identify or worse, ANOVA to calculate. And last but certainly by no means least, I want to thank my very long suffering Fiancée Kim-Louise Sparks, for putting up with gross photos of decay, weird smelling samples, and for proof reading my work before anyone else does, being grumpy when the laptop doesn’t work, and with how long I have been a student for,. I couldn’t have got this far without you. So thank you to all of the above and to anyone I may have missed. It has been a hell of a ride. - 4 - Table of Contents Abstract Page 3 Acknowledgments Page 4 List of Tables Page 11 List of Figures Page 13 Chapter one: Introduction Page 18 1.1 General introduction Page 18 1.2 The process of death and decomposition of animal remains Page 19 1.3 Forensic Entomology Page 22 1.4 Case studies Page 24 1.5 Factors affecting decomposition and insect development Page 26 1.6 Forensic entomotoxicology Page 34 1.7 Toxins other than drugs of abuse Page 40 1.8 Nicotine and related compounds Page 45 1.9 Aims Objectives and Outcomes Page 53 Chapter two: Baseline Studies Page 55 2.1 Introduction and rationale Page 55 2.2 Site descriptions Page 57 2.3 Methodology Page 58 2.3.1 Curation of specimens Page 60 2.4 Results Page 62 2.5 Discussion Page 66 2.5.1 Diptera: Sciaridae Page 66 - 5 - 2.5.2 Phoridae Page 66 2.5.3 Psychodidae Page 67 2.5.4 Heleomyzidae Page 67 2.5.5. Sepsidae Page 68 2.5.6 Dryomyzidae Page 68 2.5.7. Sphaeroceridae Page 68 2.5.8. Clussidae Page 69 2.5.9. Muscidae Page 69 2.5.9 Calliphoridae Page 70 2.5.10 Coleoptera: Carabidae Page 70 2.5.11 Leiodidae Page 70 2.5.12 Hydrophilidae Page 71 2.5.13 Histeridae Page 71 2.5.14 Nitidulidae Page 71 2.5.15 Stapylinidae Page 72 2.5.16 Hymenoptera: Braconidae Page 72 2.5.17 Vespidae Page 73 2.5.18 Lepidoptera: Tineidae Page 74 2.5.19 Hemiptera: Lygaeidae Page 74 2.5.20 Dermaptera: Forficulidae Page 74 2.5.21 Acari Page 75 2.6 Conclusion and final site selection Page 77 Chapter three: Autumn/winter pig carrion field tests Page 79 3.1 Introduction Page 79 3.2 Methodology Page 79 - 6 - 3.3 Results Page 83 3.3.1 Meteorological measurements Page 83 3.3.2 Observations on carrion Page 90 3.4 Discussion Page 93 3.4.1 Diptera: Calliphoridae Page 93 3.4.2 Syrphidae Page 95 3.4.3 Stratiomyidae Page 97 3.4.4 Fanniidae Page 98 3.4.5 Coleoptera: Silphidae Page 99 3.4.6. Staphylinidae Page 102 3.4.7 Scarabaeidae Page 106 3.4.8. Ptiliidae Page 107 3.4.9. Lathridiidae Page 109 3.4.10 Mycetophagidae Page 110 3.4.11. Nitidulidae Page 112 3.4.12. Pseudoscorpionida: Neobisiidae Page 113 3.5 Conclusion Page 114 Chapter 4: Spring/summer pig carrion field tests Page 117 4.1 Introduction Page 117 4.2 Methodology Page 118 4.3 Results Page 119 4.3.1 Observations Page 124 4.3.2 Comparative seasonal data Page 128 4.4 Discussion Page 131 4.4.1 Coleoptera: Silphidae Page 134 - 7 - 4.4.2 Scarabaeidae Page 136 4.4.3 Nitidulidae Page 136 4.4.4 Histeridae Page 137 4.4.5 Diptera: Bibionidae Page 138 4.4.6 Sphaeroceridae Page 139 4.4.7 Phoridae Page 140 4.4.8 Muscidae Page 140 4.4.9 Fanniidae Page 142 4.5 Conclusion Page 144 Chapter 5: Laboratory based element Page 145 5.1: Introduction Page 145 5.1.1 Rationale Page 145 5.1.2 The use of primary invertebrate colonisers in medico- Page 146 legal forensic entomology.