The blowflies (Diptera: Calliphoridae) include some of the world‘s most economically significant parasites of livestock. The defining characteristic of blowflies is the need for their larval stages to feed on a proteinaceous substrate, often including the tissues of a living vertebrate host, a process known as myiasis. While the evolution of myiasis has been linked to the development of key adaptations in behaviour and physiology (Stevens et al., 2006), patterns of blowfly evolution suggest that parasitism evolved independently in different blowfly groups after periods of geographic isolation (Stevens et al., 2006). However, understanding the origin and evolution of myiasis in Calliphoridae is restricted by a lack of agreed theories of evolutionary relationships and taxonomic classification (Stevens, 2003). Mitochondrial genes are some of the most widely used molecular markers in insect systematics, yet most studies have utilised only single genes, with few having systematically assessed which if any are best suited for studying particular insect orders. Accordingly, this thesis presents a comprehensive analysis of 62 hexapod mitochondrial genomes, including 55 from Insecta, and assesses the ability of mitochondrial genes to recover currently recognised insect orders as monophyletic groupings. The greatest amount of phylogenetic signal was recovered when all mitochondrial genes were analysed together, regardless of optimality criterion used (PhyML, RaxML, MrBayes). Of the single-gene analyses, COX1 out-performed all other genes, even performing as well as a combined-gene analysis under Bayesian inference. In view of this finding, nucleotide sequence data from COX1 (mitochondrial protein-coding), EF-1α (nuclear protein-coding gene), and 28S (nuclear rRNA) were combined to present one of the most comprehensive multi-gene phylogenetic studies of Calliphoridae to date, resolving many ambiguous relationships, and also including several taxa that have not previously been analysed in molecular phylogenetic studies. Within Calliphoridae, Cochliomyia hominivorax (New World screwworm fly), is widely considered one of the most destructive insect parasites of livestock in the Western hemisphere. While successful eradication programmes using sterile insect technique (SIT) have been completed in North and Central America, and on some Caribbean islands, in some areas SIT has failed. It has been hypothesized that failure of SIT may be related to genetic differentiation between populations of C. hominivorax. Consequently, intra-specific variation using nucleotide sequence data from both mitochondrial (COX1 and 12S) and nuclear (EF-1α) markers, was explored. Phylogenetic analysis of these data confirmed some population substructuring and suggested a South American origin to all Caribbean island populations, with the exception of Cuba. In agreement with previous studies, Cuban populations appeared distinct from all other Caribbean populations; however, our findings do not support a North American origin for Cuba, as has previously been suggested. Finally, this thesis attempted to explore the relationship between antigenic proteins expressed in larvae from species displaying different forms of parasitism, and in doing so assessed the utility of such target proteins as potential candidates for species-specific vaccines and diagnostic tools. However, while this work discovered distinct antigenic profiles for different blowfly species, the ability to characterize specific antigens was fundamentally limited by an apparent lack of homologous proteins in current databases.

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2008. / Submitted by Jaqueline Oliveira (jaqueoliveiram@gmail.com) on 2008-12-16T15:33:20Z No. of bitstreams: 1 DISSERTACAO_2008_KhesllerPatriciaOlaziaName.pdf: 4643181 bytes, checksum: 7929b5a30eb0eadefed71af0b29adf98 (MD5) / Approved for entry into archive by Georgia Fernandes(georgia@bce.unb.br) on 2009-02-19T15:43:23Z (GMT) No. of bitstreams: 1 DISSERTACAO_2008_KhesllerPatriciaOlaziaName.pdf: 4643181 bytes, checksum: 7929b5a30eb0eadefed71af0b29adf98 (MD5) / Made available in DSpace on 2009-02-19T15:43:23Z (GMT). No. of bitstreams: 1 DISSERTACAO_2008_KhesllerPatriciaOlaziaName.pdf: 4643181 bytes, checksum: 7929b5a30eb0eadefed71af0b29adf98 (MD5) / Os Diptera constituem uma das quatro maiores ordens de insetos. Chrysomya megacephala (Fabricius), a mosca-das-latrinas do Oriente, pertence à família Calliphoridae e encontra-se associada a questões médico-veterinárias, sanitárias e forenses. O objetivo desse trabalho foi caracterizar os aspectos ultra-estruturais da espermiogênese e do espermatozóide na referida espécie, incluindo a anatomia dos testículos e dos órgãos de armazenamento de espermatozóides nas fêmeas – as espermatecas. Estes dados são particularmente importantes em análises taxonômicas e filogenéticas, uma vez que tais células apresentam estruturas altamente conservadas. Nesta espécie, assim como na maioria dos insetos, a espermiogênese é caracterizada pelos seguintes eventos: formação do acrossoma, alongamento nuclear com condensação da cromatina, formação do flagelo e perda do excesso de citoplasma. No final do processo de espermiogênese, o espermatozóide em C. megacephala apresenta-se longo e delgado, medindo aproximadamente 590μm de comprimento. O espermatozóide é formado por acrossoma, núcleo com cromatina densamente compactada, centríolo, adjunto do centríolo, dois derivados mitocondriais preenchidos por material paracristalino e um axonema contendo: dois microtúbulos centrais, nove duplas periféricas e nove microtúbulos acessórios intercalados por fibras densas. O sistema reprodutor interno de machos e fêmeas possui modificações morfológicas que podem ser constantes em todos os ciclórrafos. O testículo de C. megacephala consiste de um único folículo e a parede do órgão é formada por uma camada de células pigmentadas e uma camada de tecido muscular. A espermateca consiste, externamente, de uma camada de células secretoras e internamente, de uma camada de cutícula que circunda a cavidade do órgão. ________________________________________________________________________________ ABSTRACT / Diptera constitutes one of the four largest orders of insects. The Oriental latrine-fly Chrysomya megacephala (Fabricius) belongs to the family Calliphoridae and is associated with medic, veterinary, sanitary and forensic issues. The aim of this research was to characterize the utrastructural aspects of the spermiogenesis and spermatozoa of the C. megacephala, including the anatomy of testis and female sperm storage organs – the spermatheca. This data is particularly important in taxonomic and phylogenetic analysis, since in these cells their structure is highly conserved. In this species as with insects in general, the spermiogenesis is characterized by the following events: acrosome formation, nuclear elongation with chromatin condensation, flagellar formation and sheading of excess cytoplasm. At the end of spermiogenesis process, the spermatozoon in C. megacephala is a long and thin cell measuring 590 μm in length. The spermatozoon comprises acrosome, nucleus with compact dense chromatin, centriole, centriolar adjunct, two mitochondrial derivatives with paracrystalline material and axoneme with: two central microtubules, nine peripheral doublets, nine accessory microtubules and dense fibers. The internal reproductive system of the male and female has morphological modifications that may be constant in all Cyclorraphous. The testis of the C. megacephala consists of a unique follicle and the wall of the organ is formed by a layer of pigmented cells and a layer of muscular tissue. The spermatheca consists externally of a secretory cell layer, and internally of a cuticle layer witch surrounds the cavity of the organ.

Made available in DSpace on 2014-06-12T15:03:29Z (GMT). No. of bitstreams: 2 arquivo1221_1.pdf: 2792260 bytes, checksum: c6bea0e531fd1905c955313d40fd8b77 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2009 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Insetos podem ser utilizados como ferramentas para auxiliar as ciências forenses, entre outros, na estimativa do intervalo pós-morte (IPM). A decomposição é um processo cuja cronologia pode ser determinada não apenas por uma série de eventos químicos e físicos, mas também pelos grupos de insetos com nichos específicos que ocupam diferentes fases. Este trabalho objetivou caracterizar a dipterofauna associada a cadáveres humanos e nas instalações do Instituto de Medicina Legal Professor Antônio Persivo Cunha (IMLAPC), Recife. Adicionalmente, visou testar a aplicabilidade de uma fórmula simplificada para estimativa do IPM utilizando evidências entomológicas, com base na comparação com informações sobre a bionomia das espécies descritas na literatura. A coleta ocorreu de setembro/2007 a fevereiro/2008. Larvas de insetos foram coletadas em 14 cadáveres do sexo masculino em avançado estágio de decomposição. Simultaneamente, adultos foram coletados em armadilhas instaladas nas dependências do IMLAPC. Imaturos foram criados até a emergência do adulto para identificar as espécies. Cinco espécies: Chrysomya albiceps, Chrysomya megacephala, Cochliomyia macellaria (Calliphoridae), Oxysarcodexia riograndensis e Ravinia belforti (Sarcophagidae) foram observadas colonizando os cadáveres, com prevalência de C. albiceps, presente em 64,3% dos casos. O ambiente circunvizinho, por sua vez, abriga uma fauna muito mais rica e abundante, com cinco famílias de Diptera (Muscidae, Fanniidae, Phoridae, Anthomyiidae e Stratiomyidae), além de espécimes de Coleoptera e Hymenoptera. A comparação entre os métodos médico-legais e os entomológicos evidenciou a eficácia deste último para determinar o tempo de morte já que, em somente um dos casos houve discrepância dos IPM obtidos. Os resultados contribuem para expandir o conhecimento sobre a diversidade de insetos necrófagos em Pernambuco e oferecem um ponto de partida para os profissionais da área jurídica e criminal

No description available.

A infestação por larvas de dípteros, conhecida como miíase, é um grande problema na pecuária em todo o mundo e pode causar perdas econômicas severas. A superfamília Oestroidea é um modelo interessante para estudar a evolução de moscas causadoras de miíase devido à diversidade de estratégias de parasitismo entre espécies intimamente relacionadas nessa família. Essas moscas são classificadas pelo seu hábito alimentar como parasitas saprófagos, obrigatórios ou parasitas facultativos. Os parasitas, em particular, podem ser subdivididos pela manifestação clínica/local de infestação como dérmica, nasofaríngea, traumática/ferida e furuncular. Com tal diversidade de estratégias, postula-se que espécies intimamente relacionadas tenham diferenças genéticas que desempenham um papel na formação desses hábitos. Aqui, utilizamos dados de expressão gênica e as sequências codificantes em escala genômica de cinco espécies (Cochliomyia hominivorax, Chrysomya megacephala, Lucilia cuprina, Dermatobia hominis e Oestrus ovis) para encontrar genes que possam estar envolvidos em diferentes estratégias e/ou preferências alimentares. Nós testamos se os 1.287 ortólogos identificados possuiam expressão diferente e restrições evolutivas em diferentes cenários. Ao comparar seus perfis de expressão gênica, encontramos dois genes regulados positivamente; um genes em espécies que causam miíase dérmica envolvido no transporte metabolização de ferro (Ferritina) e outro gene em espécies que causam miíase traumática que responde a níveis reduzidos de oxigênio (anoxia up-regulation-like). Nossa análise evolutiva mostrou um resultado semelhante. Em Ch. hominivorax, encontramos genes diferentes, mas envolvidos nas mesmas funções que podem estar evoluindo sob seleção positiva. Este é o primeiro passo para entender as origens e a evolução da diversidade de estratégias parasitárias em Oestroidea / The infestation by dipterous larvae, known as myiasis, is a major problem in livestock worldwide and can cause severe economic losses. The Oestroidea superfamily is an interesting model to study the evolution of myiasis-causing flies because of the diversity of parasitism strategies among closely-related species in this family. These flies are classified by their feeding habit as saprophagous, obligate parasites or facultative parasites. The parasites in particular can be subdivided into dermal, nasopharyngeal, traumatic/wound and furuncular. With such a diversity of parasitic strategies, we expect that closely-related species have genetic differences that play a role in shaping these habits. Here, we used gene expression and coding sequence data from five species (Cochliomyia hominivorax, Chrysomya megacephala, Lucilia cuprina, Dermatobia hominis and Oestrus ovis) to find genes that may be involved in different parasitic strategies. We tested whether 1,287 orthologs have different expression and evolutionary constrains in different scenarios. By comparing their gene expression profiles, we found two up-regulated genes; one in species causing dermic myiasis that is involved in iron transportation/metabolization (Ferritin), and other in species causing traumatic myiasis that responds to reduced oxygen levels (anoxia up- regulated-like). Our evolutionary analysis showed a similar result. In the Ch. Hominivorax branch, we found different genes, but involved in the same functions that may be evolving under positive selection. This is the first step towards understanding the origins and evolution of the diversity of parasitic strategies in Oestroidea

Les larves de Diptères nécrophages se développant sur un cadavre font face à de fortes pressions de sélection. Nous démontrons comment cet environnement extrême aurait favorisé l’apparition de stratégies comportementales efficaces et originales, basées sur des mécanismes comme la régulation thermique mais également la socialité.Ce travail pose en premier lieu les bases du comportement de régulation thermique des larves de Diptères Calliphoridae. En effet, celles-ci sont confrontées à un environnement thermique très hétérogène, dans lequel elles vont sélectionner la zone la plus appropriée à leur activité métabolique. Bien que différentes espèces exploitent la même ressource au même moment, nous avons observé que les larves de Lucilia sericata, Calliphora vomitoria et Calliphora vicina ont chacune une température préférentielle de développement. De plus, nous démontrons que ces larves sont en recherche constante de leur température préférentielle (thermorégulation), et qu’elles adaptent leur alimentation à la température du milieu. Ce premier volet d’expérimentations illustre ainsi le rôle prépondérant de la température dans le comportement des larves. En second lieu, ce travail s'intéresse à la dimension sociale des larves nécrophages, et plus particulièrement au comportement d'agrégation. Nous avons démontré chez Lucilia sericata un fort effet attractif et rétentif des congénères, rendant manifeste une prévalence de la socialité sur la régulation thermique. Les résultats sont cependant drastiquement différents dans des conditions hétérospécifiques, où la formation du groupe varie selon les températures préférentielles et les cinétiques d’agrégation de chaque espèce. Ainsi, la température sélectionnée par un groupe hétérospécifique émerge d'un compromis entre les comportements de thermorégulation et d'agrégation. Enfin, ce travail analyse l’effet de ces stratégies comportementales sur le développement des individus.Nous montrons que le comportement de thermorégulation et l’action des congénères affectent la température sélectionnée par les larves, et donc, leur développement. De tels résultats démontrent l’existence de véritables stratégies comportementales individuelles et collectives de développement, reposant sur l’optimisation de paramètres multiples permettant aux larves de se développer au mieux dans cet écosystème extrême qu’est le cadavre en décomposition. / On a cadaver, necrophagous dipteran larvae suffer from strong selection pressures during their development. The premise of this thesis is that such an extreme, competitive and constraining environment would have favored the emergence of efficient developmental strategies, based on mechanisms such as thermal regulation but also sociality. This PhD work is divided into three parts. The first part focuses on the thermal behavior of Calliphorids larvae, which are confronted with a heterogeneous thermic environment on the corps, in which they select the most appropriate area for their metabolic activity. Firstly, this part shows that larvae have a preferential developmental temperature, which is different according to the species (Lucilia sericata, Calliphora vomitoria and Calliphora vicina), although they exploit the same resource at the same time. Secondly, this part demonstrates that the larvae are always in search of this preferential temperature and thirdly, that they adapt both their displacement and their food intake according to the temperature of the nourishing substrate. This first part of experiments demonstrates that the temperature parameters have a strong effect on larval behavior. The second part of this work focuses on the social dimension of larval behavior by analyzing the influence of congeners, mainly through their active aggregation behavior. We show for Lucilia sericata a strong attractive and retentive effect of the group, making obvious that sociality prevails over thermal regulation. However, these results are radically different under heterospecific conditions where the group formation strongly depends on preferential temperatures as well as aggregation kinetics of each species. Finally, the third part of this work analyzes the effects of temperature and congeners on the development of individuals and shows that both the behavior of thermoregulation and the action of congeners impact the temperature selected by larvae, and therefore, their development. These results indicate the existence of individual and collective behavioral development strategies based on the optimization of multiple parameters that allow larvae to develop ideally in this extreme ecosystem of a decaying corpse.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-06-15T14:13:09Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Roberta Luiza Salgado Oliveira BCentral.pdf: 1516470 bytes, checksum: 15f8bd93e354db3b3b15a90212d5290b (MD5) / Made available in DSpace on 2016-06-15T14:13:09Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Roberta Luiza Salgado Oliveira BCentral.pdf: 1516470 bytes, checksum: 15f8bd93e354db3b3b15a90212d5290b (MD5) Previous issue date: 2012-02-28 / FACEPE / A fauna encontrada em carcaças animais é composta por pequenos invertebrados, que possuem a necessidade de descobrir, colonizar e usufruir rapidamente deste substrato. Este trabalho descreve o primeiro inventário sistematizado de dípteros necrófagos em um ambiente de Caatinga, bioma exclusivamente brasileiro e sujeito a temperaturas altas, umidade e pluviosidade baixa. Utilizando como modelo três carcaças de suínos, 15 kg cada um, depositada dentro de uma armadilha modificada de Shannon. As coletas foram realizadas em uma área conservada de Caatinga no município de Serra talhada, estado de Pernambuco, em abril e maio/2010. Insetos adultos foram coletados diariamente utilizando com redes entomológicas, do primeiro dia depois da morte até a completa esqueletização da carcaça. A decomposição foi dividida em cinco fases: fresca (1 dia), inchada (2 dias), decomposição ativa (4 dias), decomposição avançada (5 dias) e seco (14 dias). Foram coletadas 19, 736 adultos pertecentes a 20 familias o qual dez possuem registro prévio de necrofagia, abrangendo 57% do total de abundância e 59% do total de riqueza. A família necrófaga mais abundante foi Calliphoridae (47,3% dos espécimes), Sarcophagidae (20,8%) e Muscidae (15,5%). As espécies mais abundantes com hábitos necrófagos foram Cochliomyia macellaria (25,4% do total de indivíduos necrófagos), Chrysomya albiceps (22,3%) (Calliphoridae) e Musca domestica (15,1%) (Muscidae). Embora estágios iniciais de decomposição sejam associados com alta abundancia de indivíduos necrófagos, uma lata riqueza foi registrada nos estágios finais. A abundância de C. macellaria e Chloroprocta idioidea parecem estar associada com uma preferência por ambiente selvagem. Relatamos pela primeira vez a presença da espécie invasora C. albiceps na Caatinga, embora sua proporção na assembléia não foi tão dominante como registrado para outras áreas neotropicais. É possível inferir que, apesar das duras condições ambientais na Caatinga, o bioma abriga uma rica assembléia de dípteros necrófagos que desempenham um papel fundamental na decomposição da matéria animal. / The fauna found in animal carcasses consists of small invertebrates, which have the need to discover, colonize and rapidly take advantage of this substrate. This paper describes the first systematic inventory of necrophagous Diptera in an environment of Caatinga biome of Brazil and exclusively subjected to high temperatures, low humidity and rainfall. Using as a model three carcasses of pigs, 15 kg each, placed inside a modified Shannon trap. Field trials were performed in a preserved area of caatinga in the municipality of Serra Talhada, State of Pernambuco, in April and May/2010. Adult insects were collected daily using entomological nets, from the first day after death until the complete skeletonization of the carcass. Decomposition was divided into five stages: fresh (1 day), bloated (2 days), active decay (4 days), advanced decay (5 days) and dry (14 days). We collected 19.736 adults belonging to 20 families of which 10 have previous register necrophagy, and make up 57% of total abundance and 59% of total richness. The most abundant necrophagous families were Calliphoridae (47,3% of specimens), Sarcophagidae (20,8%) and Muscidae (15,5%). The most abundant species with necrophagous habits were Cochliomyia macellaria (25,4% of total necrophagous individuals), Chrysomya albiceps (22,3%) (Calliphoridae) and Musca domestica (15,1%) (Muscidae). Although early stages of decomposition were associated with higher abundance of necrophaguos individuals, a higher richness was registered at the final stages. The abundance of C. macellaria and Chloroprocta idioidea seems to be associated with a preference for wild environment. We report for the first time the presence of the invasive species C. albiceps in the caatinga, although its proportion in the assemblage was not as dominant as registered for other Neotropical areas. It is possible to infer that despite the harsh environmental conditions at the Caatinga, this biome harbours a rich assemblage of necrophagous Dipterans that play a key role in animal matter decomposition.

<p>In terms of forensic entomology, one area that is scrutinized most is the estimation of a minimum post mortem interval (mPMI) based on insects that are present at a crime scene. The identification of the insects found at the scene if the first step to calculate a mPMI. However, currently there are no methods that can present the courts with accurate statistical error rates in identification, because the current methods are reliant on an expert’s use of a morphological key to identify the specimen, and this identification method does not produce a confidence value. This project aimed to test a method of identification using geometric morphometrics that can produce confidence intervals to provide to the courtrooms. </p><p> Before any identification could start, a standard preservation protocol was developed to ensure that all diagnostic features are preserved, and specimens can be identified in the same way. A clearing method was designed to clear specimens within 24hrs using potassium hydroxide, so they can be dissected and mounted the next day. The dissection of the specimens was a simple six-step procedure to split the mouth hooks, the cuticle and the posterior spiracle. This procedure ensures that all diagnostic features are preserved on a microscope slide.</p><p> With all of the features preserved, the microscope slide is photographed for storage and an investigator can perform geometric morphometrics to identify the insect. This study tested the application of geometric morphometrics to distinguish between three genera of Calliphoridae (<i>Calliphora</i>,<i>Lucilia</i>, <i>Phormia</i>), from three locations in the US (Delaware, Indiana, California). Results showed significant (p-value: <0.05) variation in shape among all genera. When genera were tested for shape differences based on location, these variations were also significant (p-value: <0.05). The implication of these results is that enough shape difference exists to distinguish between these genera and to distinguish between populations.</p>

Population genetic structure is difficult to assess in blow flies (Diptera: Calliphoridae) due to high connectivity and genetic diversity of subpopulations. Previous studies revealed high relatedness among individuals within wild samples of blow fly populations, however broad geographic structure was absent. The aim of this research was to determine if blow fly genetic structure exists at a fine spatiotemporal resolution and, if so, to elucidate the influence of environmental factors and resource availability on fly genetics. Specifically, blow fly population genetic patterns were tested against anull hypothesis that flies adhere to a patchy population model with high genetic diversity (i.e. no structure) and high resource availability. Samples of the black blow fly, <i>Phormia regina</i> Meigen (Diptera: Calliphoridae), were collected at six urban parks in Indiana, USA (=urban) in 2016 and 2017 (N = 14 and 16 timepoints, respectively). Additional sampling in different ecoregions was performed to determine if trends observed at a high-resolution scale were also present at a broad geographic scale. Therefore, <i>P. regina</i> were also collected at four sites within two national parks (the Great Smoky Mountains and Yellowstone National Parks) over a three-day period. Randomly selected females (N = 10) from each sample underwent the following analyses: 1) gut DNA extraction, 2) molecular analysis at 6 microsatellite loci, 3) vertebrate-specific 12S and 16S rRNA sequencing, and, 4) vertebrate fecal metabolite screening. Flies from the national parks and a comparable subset of urban data also underwent stable isotopeanalysis (SIA) to determine larval food source. Overall, strong seasonal population genetic structure was observed over both years in the urban environment (2016 F’_ST= 0.47, 2017 F’_ST0.34), however spatial structure was lacking, as seen in previous studies (2016 F’_ST= 0.04, 2017 F’_ST0.03). Weather conditions prior to and on the day of blow fly collections, interspecific competition, and resource availability greatly impacted the genetic diversity and kinship of <i>P. regina</i>. A total of 17 and 19 vertebrate species were detected by flies in 2016 and 2017, respectively, and many flies tested positive for vertebrate feces, suggesting that many varied resources are important for maintaining high gene flow among geographic locations. Genetic diversity was non-existent in flies collected from the Smokies (F’_ST= 0.00), while very slight spatial structure existed in the Yellowstone populations (F’_ST= 0.07). Environmental factors such as temperature, humidity, and wind speed were all statistically relevant in maximizing fly collections with vertebrate resources. In 720 min of total sampling time in the national parks and a subset of urban data, 28 vertebrate species were identified, and fecal resources appeared to be the most abundant in Yellowstone. Stable isotopeanalysis revealed a majority of larval resources in the national parks were herbivores, with a more even distribution of carnivore and herbivore carcasses present in the urban environment, which likely explains the high genetic diversity of adult flies in these regions. Overall, the null hypothesis that <i>P. regina</i> adheres to a patchy population model could not be rejected for the Smokies populations. However, the urban and Yellowstone populations appear to adhere to a Levins metapopulation model in which variable availability in resources leads to random bottleneck events in the local populations. Overall, environmental conditions, competition, and resource availability are all important factors influencing <i>P. regina </i>population genetic structure in different environments.

Blow flies (Diptera: Calliphoridae) are among the most dominant and conspicuous insects in the decomposition process. They are important in forensic entomology to determine time of death and, in certain situations, cause of death or relocation of a body. Insects are now included as standard operating procedures in crime scene investigations in many countries, however, this is not standard procedure in the Caribbean area due to lack of knowledge of insects involved in cadaveric decomposition. Successful application of forensic entomology depends on solid underlying data. Our main goal is to advance the knowledge of Calliphoridae in the Caribbean to enable forensic entomology studies. We performed a mega-transect across the Caribbean and extensively collected flies attracted to rotten meat baits during five years from 2011 to 2015. Overall we collected 61,332 flies of which 34,650 were Calliphoridae. We sampled 16 of the 18 species of forensically important Caribbean Calliphoridae and three continental species. We determine the diversity and distribution of Calliphoridae in the Caribbean. We also present a thorough DNA barcode dataset, covering the geographic range of most species in the region. Finally we established phylogenetic relationships among Calliphoridae species and test biogeographical hypotheses, and patterns of diversification and endemism in the Caribbean. In sum, this is the most comprehensive study of the family Calliphoridae from the Caribbean that will open the door for future research on forensic entomology in the region.