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Insect Olfaction As an Information Filter for Chemo-Analytical Applications

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Insect olfaction as an information filter for chemo-analytical applications Dissertation In partial fulfillment of the requirements for the degree of Doctor of Philosophy (PhD) of the Faculty of Forest Science and Forest Ecology Georg-August-University Göttingen Submitted by Sebastian Paczkowski, born in Berlin-Schöneberg Göttingen, 2013 1. Reviewer: Prof. Dr. Stefan Schütz 2. Reviewer: Prof. Dr. Ursula Kües Date of the oral examination: 29.05.2013 Acknowledgements I like to thank all the people who advised and supported my work at the department of forest zoology and forest protection. I also like to thank my supervisor, Prof. Dr. Stefan Schütz, for his support and advice. I further like to thank Prof. Dr. Ursula Kües for her supervision of this thesis and Prof. Dr. Edmone Roffael for the evaluation of my oral examination. I like to thank especially Dr. Bernhard Weissbecker and Dr. Gerrit Holighaus for their help and their critical opinions during so many discussions. Further thanks go to all of the present and former staff of the department of forest zoology and forest protection. Thank you very much: Dr. Kai Füldner, Prof. Dr. Sergio Angeli, Evelin Kistner, Brunhilde Brunotte, Dr. Prodepran Thakeow, Dr. Bettina Johne, Julian Heiermann, Dr. Sonja Weißsteiner, Christine Rachow, Jan Seelig, Dr. Maximilian von Fragstein und Niemstorff, Dr. Martin Gabriel, Sara Nicke, Friederike Maibaum, Kira Duntemann, Ulrike Eisenwiener, Jörg Berger, Ullrich Aschmann, Reinhold Dankworth, Dr. Ferdinant Rühe, and Roswita Walbrecht. I further like to thank all the external persons who supported my work: Prof. Dr. Claus-Dieter Kohl, Dr. Tilmann Sauerwald, Prof. Dr. Helmut Schmitz, Dr. Redelf Kraft, Dr. Jörg Gottschald, Moritz Essinger, Marco Bauer, Alexander Weiß, Daniel Meixner, Arnim Titze, Heino Schmeiß, Brian Inglis, and Prof. Dr. Barbara Wilson. Further thanks go to all of the students who I supervised and who thereby contributed to my work. I like to thank the Cusanuswerk, Bischöfliche Studienstiftung, for the financial support. I like to thank my wife, Dr. Marta Paczkowska and my children Oliver Pavel, Isabella Jadwiga and Katharina Lotta for giving sense to my life. Zusammenfassung Die Geruchswahrnehmung von Insekten ist an die spezifischen Anforderungen der Ökosysteme angepasst, in denen sie leben. Von Insekten wahrgenommene Volatile können Informationen über den Ort eines geeigneten Ovipositionsplatzes oder den physiologischen Zustand einer Wirtspflanze geben. Da der Geruchssinn der Insekten sich seit Millionen von Jahren an die Wahrnehmung bestimmter Volatile angepasst hat, können die wahrgenommenen Volatile als Markerstoffe angesehen werden, die zuverlässig einen Ökosystemzustand angeben können. Die Identifikation dieser Markerzustände und die Evaluation ihrer Nützlichkeit für spurenanalytische Anwendungen ist das Ziel dieser Arbeit. Es wurden drei Themen ausgewählt um die Verwendungsmöglichkeit des Insektengeruchssinns für Sensoranwendungen zu überprüfen: Fleischfrische, Todeszeitbestimmung (post mortem Intervall, PMI) und frühzeitige Feuererkennung. Spurenanalytik (GC-MS), Elektrophysiologie (EAG, GC- MS/EAD), Verhaltensversuche und Feldtests wurden angewandt, um die chemoökologische Interaktion der Schmeißfliege Calliphora vicina ud der „Feuerkäfer Melanophila cuspidata, Merimna atrata and Acanthocnemus nigricans mit ihrer natürlichen Umgebung zu untersuchen. Die Resultate aus diesen Methoden erlaubten die Selektion von Volatilen, die drei Kriteria erfüllen: hohe Quantität, zuverlässige Emission und die ausschliessliche Emission von der untersuchten und keiner anderen Quelle. Diese drei Auswahlkriterien wurden aufgestellt um zu gewährleisten, dass die ausgewählten Markervolatile nicht nur zuverlässig mit dem physiologischen Status der Substrate (Alter von verderbendem Fleisch, PMI, Temperatur von erhitzten Spänen) korrelieren, sondern auch den Ansprüchen technischen Sensorsysteme genügen. Im Falle des alternden Fleisches unter warmen und trockenen Bedingungen ist Nonanal ein Zeiger für die korrekte Reifung des Fleisches zu Schinken. Dimethyl Trisulfid, Phenol und Indol zeigen das Verderben des Fleisches unter warmen und feuchten Bedingungen an. Bei niedrigen Temperaturen sind zunehmende 2,3 Butandiol Emission und abnehmende Nonanal Emission ein Zeiger für zunehmendes Verderben des Fleisches, sowohl unter trockenen als auch feuchten Bedingungen. Allerdings wurde 2,3 Butandiol nicht von C. vicina wahrgenommen, da die Fliege während der Vegetationszeit aktiv ist und unter diesen Bedingungen Dimethyl Trisulfid, Phenol und Indol emittiert werden. Nonanal, Hexanal, Dimethyl Disulfid, Dimethyl Trisulfid, Butan-1-ol und Phenol wurden als nützliche Volatile zur Eingrenzung der Leichenliegezeit ausgewählt. Die genannten Aldehyde sind typisch für die frühen Stadien der Verwesung, gefolgt von Butan-1-ol und den Schwefelsulfiden. Phenol wird hauptsächlich in den späteren Verwesungsstadien emittiert. Allerdings werden Phenol und 1-Butanol nicht von der Fliege wahrgenommen, da diese ein Generalist ist und nicht zwischen dem geblähten und aktivem Stadium der Verwesung unterscheiden muss. Daher wird zusätzlich die Geruchswahrnehmung des Speckkäfers Dermestes maculans betrachtet. Die Untersuchung anderer necrophager Insekten können weitere Einblicke in die Zeitabhängigkeit der Volatilemissionen während der Vertebratenverwesung geben. Die im Rahmen dieser Arbeit gewonnenen Ergebnisse erlauben eine Korrelation zwischen Emissionen und Leichenliegezeit und können eine Grundlage für eine volatilenbasierte post mortem Zeitbestimmung für Polizeikräfte sein. Terpene, aliphatische Aldehyde, Furfural und Methoxyphenole werden von erhitztem Holz emittiert. Komponenten aus all diesen Stoffklassen werden von M. cuspidata und M. atrata wahrgenommen, da diese Käfer den Erhitzungszustand von Baumstämmen nach einem Feuer erkennen können, um einen geeigneten Ovipositionsplatz zu finden. A. nigricans nimmt vor allem Methoxyphenole wahr, wohingegen M. atrata und M. acuminata Furfural zur Orientierung verwenden. Diese Unterschiede komplementieren das bekannte Verhalten dieser Käfer in ihrer natürlichen Umgebung, da A. nigricans auf verkohlten Stämmen, die große Mengen von Methoxyphenolen emittieren, seine Eier ablegt, und M. atrata und M. cuspidata auf Stämmen ihre Eier ablegen, die vom Feuer zwar äußerlich verkohlt, aber innerlich nur erhitzt wurden und daher Furfural emittieren. Halbleitergassensoren, die Furfural mit einer ausreichenden Selektivität vermessen können, können Holzbrände schon während ansteigender Temperatur und damit vor der Entzündung detektieren. Derartige Sensoren können sowohl als neuartige Frühwarnsysteme für die holzverarbeitende Industrie als auch für Waldbrandwarnsysteme oder den Haushalt dienen. Abstract The olfaction of insect species is adapted to the requirements of the specific ecosystem they are living in. Volatiles perceived by insects can transport information on the location of an oviposition place or the physiological status of a host plant. As the olfaction of insects has adapted to the perception of distinct volatiles since millions of years, these volatiles can be regarded as marker compounds, which enable a reliable assessment of an ecological status. The identification of such marker compounds and the evaluation of their usefulness for trace analytical applications is the aim of this thesis. Three topics were selected to evaluate the utilization of insect olfaction for sensor applications: Meat spoilage, post mortem interval (PMI) estimation, and the volatile based detection of wood fire before ignition. Trace analysis (GC-MS), electrophysiology (EAG, GC-MS/EAD), behavioral studies and field tests were used to understand the volatile based interaction of the blowfly Calliphora vicina and the fire beetles Melanophila cuspidata, Merimna atrata, and Acanthocnemus nigricans with their environment. The results gained by these methods allowed the selection of volatile marker compounds that fulfilled three criteria: high abundance, reliable emission, and unique emission from the substrate and not from any other sources. These criteria should ensure that the marker compounds`s emission is not only reliably correlated to a physiological state of the substrate, e.g. an increasing spoilage of meat, a time point after death of a vertebrate, or a distinct temperature of wood prior to ignition, but that the marker compound can be sensed by technical sensor systems, as well. In case of meat aging, the predominant emission of nonanal under warm conditions was an indicator for the correct maturation of the meat to ham. Dimethyl trisulfide, phenol, and indole indicated moisture induced spoilage under warm conditions. Under cold conditions, increasing 2,3 butanediol emission and decreasing nonanal emission was a spoilage indicator for both the dry and the wet samples. However, 2,3 butanediol was not perceived by the fly, because its activity is restricted to the warm vegetation period, where dimethyl trisulfide, phenol, and indole are emitted. Nonanal, hexanal, dimethyl disulfide, dimethyl trisulide, butan-1-ol, and phenol are useful volatiles to estimate the PMI of vertebrates. The aldehydes are emitted in the early stages of vertebrate decay, followed by butan-1-ol and the sulfur compounds. Phenol is predominantly emitted in the later stages of decay. However, butan-1-ol and phenol are not perceived by the fly, which is a generalist among carrion visitors and does not differentiate between the early stage, the bloated stage, and the active
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  • Evaluating the Effects of Temperature on Larval Calliphora Vomitoria (Diptera: Calliphoridae) Consumption

    Evaluating the Effects of Temperature on Larval Calliphora Vomitoria (Diptera: Calliphoridae) Consumption

    Evaluating the effects of temperature on larval Calliphora vomitoria (Diptera: Calliphoridae) consumption Kadeja Evans and Kaleigh Aaron Edited by Steven J. Richardson Abstract: Calliphora vomitoria (Diptera: Calliphoridae) are responsible for more cases of myiasis than any other arthropods. Several species of blowfly, including Cochliomyia hominivorax and Cocholiomya macellaria, parasitize living organisms by feeding on healthy tissues. Medical professionals have taken advantage of myiatic flies, Lucillia sericata, through debridement or maggot therapy in patients with necrotic tissue. This experiment analyzes how temperature influences blue bottle fly, Calliphora vomitoria. consumption of beef liver. After rearing an egg mass into first larval instars, ten maggots were placed into four containers making a total of forty maggots. One container was exposed to a range of temperatures between 18°C and 25°C at varying intervals. The remaining three containers were placed into homemade incubators at constant temperatures of 21°C, 27°C and 33°C respectively. Beef liver was placed into each container and weighed after each group pupation. The mass of liver consumed and the time until pupation was recorded. Three trials revealed that as temperature increased, the average rate of consumption per larva also increased. The larval group maintained at 33°C had the highest consumption with the shortest feeding duration, while the group at 21°C had lower liver consumption in the longest feeding period. Research in this experiment was conducted to understand the optimal temperature at which larval consumption is maximized whether in clinical instances for debridement or in myiasis cases. Keywords: Calliphora vomitoria, Calliphoridae, myiasis, consumption, debridement As an organism begins to decompose, target open wounds or necrotic tissue.