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Item Blow fly stable isotopes reveal larval diet: A case study in community level anthropogenic effects(PLOS, 2021-04) Owings, Charity G.; Gilhooly, William P. III; Picard, Christine J.; Earth Sciences, School of ScienceResponse to human impacts on the environment are typically initiated too late to remediate negative consequences. We present the novel use of stable isotope analysis (SIA) of blow flies to determine human influences on vertebrate communities in a range of human-inhabited environments, from a pristine national park to a dense metropolitan area. The refrain “you are what you eat” applies to the dietary isotope record of all living organisms, and for carrion-breeding blow flies, this translates to the type of carcasses present in an environment. Specifically, we show that carnivore carcasses make up a large proportion of the adult fly’s prior larval diet, which contrasts to what has been reportedly previously for the wild adult fly diet (which consists of mostly herbivore resources). Additionally, we reveal the potential impact of human food on carcasses that were fed on by blow flies, underscoring the human influences on wild animal populations. Our results demonstrate that using SIA in conjunction with other methods (e.g., DNA analysis of flies) can reveal a comprehensive snapshot of the vertebrate community in a terrestrial ecosystem.Item Genetic and genomic selection in insects as food and feed(Wageningen Academic, 2021) Eriksson, T.; Picard, Christine J.; Biology, School of ScienceThis review will summarise existing tools and resources and highlight areas of focus for the insects as food and feed industry for the production of insects as alternative protein sources. By applying knowledge gained from other agricultural organisms coupled with the ease of insect population growth and rearing capabilities, and the increase in biotechnological advances, strains optimised for various economic and biological traits should be one of the most attainable goals for researchers and insect farmers alike. We have reviewed strengths (and weaknesses) of various genetic and genomic approaches, and consider the future of insect farming in the context of genetic and genomic selection of insects.Item Blow fly stable isotopes reveal larval diet: A case study in community level anthropogenic effects(Public Library of Science Journals, 2021-04-14) Owings, Charity G.; Gilhooly, William P., III; Picard, Christine J.; Chemistry and Chemical Biology, School of ScienceResponse to human impacts on the environment are typically initiated too late to remediate negative consequences. We present the novel use of stable isotope analysis (SIA) of blow flies to determine human influences on vertebrate communities in a range of human-inhabited environments, from a pristine national park to a dense metropolitan area. The refrain “you are what you eat” applies to the dietary isotope record of all living organisms, and for carrion-breeding blow flies, this translates to the type of carcasses present in an environment. Specifically, we show that carnivore carcasses make up a large proportion of the adult fly’s prior larval diet, which contrasts to what has been reportedly previously for the wild adult fly diet (which consists of mostly herbivore resources). Additionally, we reveal the potential impact of human food on carcasses that were fed on by blow flies, underscoring the human influences on wild animal populations. Our results demonstrate that using SIA in conjunction with other methods (e.g., DNA analysis of flies) can reveal a comprehensive snapshot of the vertebrate community in a terrestrial ecosystem.Item The yellow mealworm (Tenebrio molitor) genome: a resource for the emerging insects as food and feed industry(Wageningen, 2020) Eriksson, T.; Andere, A. A.; Kelstrup, H.; Emery, V. J.; Picard, Christine J.; Biology, School of ScienceThe yellow mealworm, the larval stage of the darkling beetle Tenebrio molitor, shows great promise as an alternative source of animal protein. Herein we present the 312 Mb draft genome assembled using 10x Genomics linked-read technology to inform research efforts and to provide resources to optimise yellow mealworm for mass production and consumption. The genome with a contig N50 of 39,478 bp contains 89% of conserved arthropod genes among the > 20,000 genes assembled (complete and partial genes). This draft assembly represents a valuable resource to understanding T. molitor biology as a means of producing alternative, sustainable protein for the growing population and in the face of changing climates.Item Female Blow Flies As Vertebrate Resource Indicators(Springer Nature, 2019-07-22) Owings, Charity G.; Banerjee, Aniruddha; Asher, Travis M. D.; Gilhooly, William P.; Tuceryan, Anais; Huffine, Mary; Skaggs, Christine L.; Adebowale, Iyun M.; Manicke, Nicholas E.; Picard, Christine J.; Biology, School of ScienceRapid vertebrate diversity evaluation is invaluable for monitoring changing ecosystems worldwide. Wild blow flies naturally recover DNA and chemical signatures from animal carcasses and feces. We demonstrate the power of blow flies as biodiversity monitors through sampling of flies in three environments with varying human influences: Indianapolis, IN and two national parks (the Great Smoky Mountains and Yellowstone). Dissected fly guts underwent vertebrate DNA sequencing (12S and 16S rRNA genes) and fecal metabolite screening. Integrated Nested Laplace Approximation (INLA) was used to determine the most important abiotic factor influencing fly-derived vertebrate richness. In 720 min total sampling time, 28 vertebrate species were identified, with 42% of flies containing vertebrate resources: 23% DNA, 5% feces, and 14% contained both. The species of blow fly used was not important for vertebrate DNA recovery, however the use of female flies versus male flies directly influenced DNA detection. Temperature was statistically relevant across environments in maximizing vertebrate detection (mean = 0.098, sd = 0.048). This method will empower ecologists to test vertebrate community ecology theories previously out of reach due practical challenges associated with traditional sampling.Item Amplified fragment length polymorphism analysis supports the valid separate species status of Lucilia caesar and L. illustris (Diptera: Calliphoridae)(Taylor & Francis:, 2017-12-08) Picard, Christine J.; Wells, Jeffrey D.; Ullyot, Anne; Rognes, Knut; Biology, School of ScienceCommon DNA-based species determination methods fail to distinguish some blow flies in the forensically and medically important genus Lucilia Robineau-Desvoidy. This is a practical problem, and it has also been interpreted as casting doubt on the validity of some morphologically defined species. An example is Lucilia illustris and L. caesar, which co-occur in Europe whilst only L. illustris has been collected in North America. Reports that these species shared both mitochondrial and nuclear gene sequences, along with claims that diagnostic morphological characters are difficult to interpret, were used to question their separate species status. We report here that amplified fragment length polymorphism profiles strongly support the validity of both species based on both assignment and phylogenetic analysis, and that traditional identification criteria based on male and female genital morphology are more reliable than has been claimed.Item New Distribution Record for Lucilia cuprina (Diptera: Calliphoridae) in Indiana, United States(Oxford University Press, 2018-07-01) Owings, Charity G.; Picard, Christine J.; Biology, School of ScienceDetermining range expansion for insect species is vital in order to evaluate their impact on new ecosystems and communities. This is particularly important for species which could be potentially harmful to humans or domestic animals. Lucilia cuprina Wiedemann (Diptera: Calliphoridae) can act as a facultative ectoparasite and has an extensive history as the primary inducer of sheep-strike in Australia, New Zealand, and Africa. We present here the first record of this species in Indiana, United States. Lucilia cuprina's range expansion northward in the United States may be indicative of changing environmental conditions conducive to the proliferation of this species into historically cooler climates. The presence of this species could significantly impact forensic death investigations utilizing dipteran larvae to estimate a minimum postmortem interval. If range expansion of this species is not taken into account by a forensic entomologist (especially if L. cuprina is not known previously in their region), an inaccurate minimum postmortem interval (PMIMIN) estimation may be made, given the differences in development times for both species. Therefore, the range expansion of this fly could have large impacts for many different entomological disciplines.Item Ubiquity of polystyrene digestion and biodegradation within yellow mealworms, larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae)(Elsevier, 2018) Yang, Shan-Shan; Wu, Wei-Min; Brandon, Anja M.; Fan, Han-Qing; Receveur, Joseph P.; Li, Yiran; Wang, Zhi-Yue; Fan, Rui; McClellan, Rebecca L.; Gao, Shu-Hong; Ning, Daliang; Phillips, Debra H.; Peng, Bo-Yu; Wang, Hongtao; Cai, Shen-Yang; Li, Ping; Cai, Wei-Wei; Ding, Ling-Yun; Yang, Jun; Zheng, Min; Ren, Jie; Zhang, Ya-Lei; Gao, Jie; Xing, Defeng; Ren, Nan-Qi; Waymouth, Robert M.; Zhou, Jizhong; Tao, Hu-Chun; Picard, Christine J.; Benbow, Mark Eric; Criddle, Craig S.; Biology, School of ScienceAcademics researchers and “citizen scientists” from 22 countries confirmed that yellow mealworms, the larvae of Tenebrio molitor Linnaeus, can survive by eating polystyrene (PS) foam. More detailed assessments of this capability for mealworms were carried out by12 sources: five from the USA, six from China, and one from Northern Ireland. All of these mealworms digested PS foam. PS mass decreased and depolymerization was observed, with appearance of lower molecular weight residuals and functional groups indicative of oxidative transformations in extracts from the frass (insect excrement). An addition of gentamycin (30 mg g−1), a bactericidal antibiotic, inhibited depolymerization, implicating the gut microbiome in the biodegradation process. Microbial community analyses demonstrated significant taxonomic shifts for mealworms fed diets of PS plus bran and PS alone. The results indicate that mealworms from diverse locations eat and metabolize PS and support the hypothesis that this capacity is independent of the geographic origin of the mealworms, and is likely ubiquitous to members of this species.Item Factors Affecting Species Identifications of Blow Fly Pupae Based upon Chemical Profiles and Multivariate Statistics(MDPI, 2017-04-11) Kranz, William; Carroll, Clinton; Dixon, Darren A.; Goodpaster, John V.; Picard, Christine J.; Chemistry and Chemical Biology, School of ScienceAlternative methods for the identification of species of blow fly pupae have been developed over the years that consist of the analyses of chemical profiles. However, the effect of biotic and abiotic factors that could influence the predictive manner for the tests have not been evaluated. The lipids of blowfly pupae (Cochliomyia macellaria, Lucilia cuprina, Lucilia sericata, and Phormia regina) were extracted in pentane, derivatized, and analyzed by total-vaporization solid phase microextraction gas chromatography-mass spectrometry (TV-SPME GC-MS). Peak areas for 26 compounds were analyzed. Here we evaluated one biotic factor (colonization) on four species of blow flies to determine how well a model produced from lipid profiles of colonized flies predicted the species of flies of offspring of wild-caught flies and found very good species identification following 10 generations of inbreeding. When we evaluated four abiotic factors in our fly rearing protocols (temperature, humidity, pupation substrate, and diet), we found that the ability to assign the chemical profile to the correct species was greatly reduced.Item Optimization of total vaporization solid-phase microextraction (TV-SPME) for the determination of lipid profiles of Phormia regina, a forensically important blow fly species(Springer, 2017-11) Kranz, William; Carroll, Clinton; Dixon, Darren; Picard, Christine J.; Goodpaster, John; Chemistry and Chemical Biology, School of ScienceA new method has been developed for the determination of fatty acids, sterols, and other lipids which naturally occur within pupae of the blow fly Phormia regina. The method relies upon liquid extraction in non-polar solvent, followed by derivatization using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) w/ 1% trimethylchlorsilane (TMCS) carried out inside the sample vial. The analysis is facilitated by total vaporization solid-phase microextraction (TV-SPME), with gas chromatography-mass spectrometry (GC-MS) serving as the instrumentation for analysis. The TV-SPME delivery technique is approximately a factor of five more sensitive than traditional liquid injection, which may alleviate the need for rotary evaporation, reconstitution, collection of high performance liquid chromatography fractions, and many of the other pre-concentration steps that are commonplace in the current literature. Furthermore, the ability to derivatize the liquid extract in a single easy step while increasing sensitivity represents an improvement over current derivatization methods. The most common lipids identified in fly pupae were various saturated and unsaturated fatty acids ranging from lauric acid (12:0) to arachinoic acid (20:4), as well as cholesterol. The concentrations of myristic acid (14:0), palmitelaidic acid (16:2), and palmitoleic acid (16:1) were the most reliable indicators of the age of the pupae.