Mejoramiento de un método no comercial para la extracción de ADN de moscas de interés médico-legal

La obtención de ADN de moscas de interés médico-legal es de relevancia para una variedad de aplicaciones. Aunque existen métodos de extracción comerciales de ADN, su uso rutinario es limitado, en algunos escenarios. En este contexto, el uso de métodos no comerciales constituye una alternativa; sin embargo, su optimización es clave para mejorar el flujo de trabajo y los resultados. Este trabajo evaluó el impacto de variaciones a un método de precipitación salina sobre la concentración y la pureza del ADN recuperado. No se encontraron diferencias significativas en la concentración de ADN extraído entre los diferentes tiempos de incubación, probados durante la fase de extracción, mientras que el incremento en el volumen de etanol absoluto, en la fase de precipitación de ADN, mejoró significativamente la concentración de ADN obtenido. Las modificaciones propuestas reducen el tiempo de ejecución y la concentración de ADN obtenido comparado con el protocolo original.

Obtaining DNA from flies of medico-legal interest is relevant for a variety of applications. Although commercial extraction methods offer optimal DNA, their routine use is limited in some settings. In this context, the use of non-commercial methods constitutes an alternative in laboratories with limited resources however, its optimization is key to improving the workflow and the results. This work evaluated the impact of variations to a saline precipitation method on the concentration and purity of the recovered DNA. No significant differences were found in the concentration of extracted DNA between the different incubation times tested during the extraction phase. In contrast, the increased volume of absolute ethanol in the DNA precipitation phase significantly improved the concentration of DNA obtained. The proposed modifications reduce the runtime and DNA concentration obtained compared with the original protocol.

New developmental data of Chrysomya megacephala (Diptera: Calliphoridae) in tropical temperatures and its implications in forensic entomology

@#The estimation of the postmortem interval (PMI) is an essential information in death investigations. It is necessary to know the developmental data of the most important necrophagous insect species in every geographical area. Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) is one of the most common species associated with human body decomposition, especially in the tropics, so a precise knowledge of its life cycle is crucial. However, despite its ubiquity, developmental data in a range of tropical temperatures is scarce. For this reason, the aim of this study is to provide the developmental data of the blowfly, C. megacephala, in a range of tropical temperatures and to apply these data in forensic entomology. Four temperatures were examined (i.e., 27.0°C 29.5°C, 32.0°C and 34.5°C) and the time of developments from egg to adult were recorded. To build the growth curves, five larvae per day were measured with a digital caliper. Accumulated degree-days were calculated and the isomorphen diagram for this species was plotted. As we predicted, the results showed that the life cycle of this species was affected by the increasing temperature. The implications of these new data for determining the minimum PMI in forensic entomology were discussed.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul - Brazil / Primeira lista de parasitoides em moscas de importância médica, veterinária e forense no Rio Grande do Sul - Brasil

Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42' S; 52º 18' W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42' S; 52º 18' O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul – Brazil / Primeira lista de parasitoides em moscas de importância médica, veterinária e forense no Rio Grande do Sul - Brasil

Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42' S; 52º 18' W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42' S; 52º 18' O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul Brazil

Abstract Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42 S; 52º 18 W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Resumo Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42 S; 52º 18 O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

Application of Sarcosaprophagous Insects to Estimate the Postmortem Interval in 11 Cases / 法医学杂志

Objective To test the feasibility and accuracy of with sarcosaprophagous insects postmortem interval （PMI） estimation with sarcosaprophagous insects and provide references for estimation practice. Methods Eleven cases confirmed by the detection results, with complete entomological evidence were selected. The insect species, estimation results and true results involved in the cases were statistically analyzed and compared. Results Thirteen species of insects were found at the criminal scene, including Chrysomya megacephala （Fabricius）, Chrysomya rufifacies （Macquart）, Chrysomya nigripes （Aubertin）, Lucilia sericata （Meigen）, Hydrotaea spinigera Stein, Muscina stabulans （Fallén）, Sarcophagid （species were not identified）, Megaselia scalaris （Loew）, Hermetia illucens （Linnaeus）, Saprinus splendens （Paykull）, Creophilus maxillosus （Linnaeus）, Dermestes maculatus （De Geer） and Necrobia ruficollis （Fabricius）. The PMI of all eleven cases was within the range of estimated PMI. The estimated results of 72.73% cases were on the same day of the true results. Conclusion Sarcosaprophagous insects can estimate the PMI simply and conveniently. In cases where the PMI is within the time range of one generation of flies or beetles, the estimation results are relatively accurate. However, the estimation is less accurate when the PMI is beyond the time range.

Species Identification of Sarcosaprophagous Flies Based on Vein Digital Image Analysis / 法医学杂志

Objective To identify species of common sarcosaprophagous flies based on digital image analysis of veins, in order to provide new idea for fast and accurate species identification of sarcosaprophagous flies in forensic entomology. Methods Random trapping of 226 male and female sarcosaprophagous flies that comprised of 7 common species, including Sarcophaga peregrina, Parasarcophaga ruficornis, Sarcophaga dux, Seniorwhitea reciproca, Bercaea cruentata, Aldrichina grahami, and Synthesiomysia nudiseta with carrion in the field was conducted. The 17 landmarks on the right wing of each fly were digitally processed and the images were analyzed. The effects of allometry were evaluated using a permutation test. Wing shape variations among 7 sarcosaprophagous fly species and female species was analyzed using canonical variate analysis （CVA）. Additionally, cross-validation test was used to evaluate the reliability of classification. Results Among 7 sarcosaprophagous fly species and female species, the effect of allometry had statistical significance （P<0.05）. The CVA results showed that among 7 sarcosaprophagous fly species and female species, differences in the wing shape were significant, and the first two canonical variates accounted for 82.9% and 84.1% of the total variation of vein shape. Vein digital image analysis can be used to separate the 7 common sarcosaprophagous flies, with an overall species identification accuracy of 81.2%-100.0%, and with a species identification accuracy of 75.0%-100.0% to distinguish the female flies of the 7 sarcosaprophagous flies species. Conclusion Vein digital image analysis is a relatively convenient and reliable method for identification of insect species, which can be used for species identification of common sarcosaprophagous flies.

Transcriptome Sequencing Analysis of Chrysomyia Megacephala Pupae in Different Growing Periods / 法医学杂志

Objective To study the growth regulation, environmental adaption and epigenetic regulation of Chrysomyia Megacephala pupae, in order to obtain the transcriptome data of Chrysomyia Megacephala in different growing periods, and lay the foundation for forensic application. Methods The Chrysomyia Megacephala was cultivated and after pupation, 3 pupae were collected every 24 h from pupation to emergence, and stored at -80 ℃ for later use. High-throughput sequencing was performed by Illumina Hiseq 4000 and Unigenes were obtained. The Unigenes were compared by comparison tool BLAST from NCBI in databases such as NR, STRING, SWISS-PROT （including Pfam）, GO, COG, KEGG in order to obtain the corresponding annotation information. The expression amount of Unigenes obtained by sequencing in Chrysomyia Megacephala in six different growing periods was calculated by FPKM method, and the discrepant genes were screened according to the following standards： the log2 multiple absolute value of FPKM expression amount between two different growing periods must be larger than 1 （log2|FC|>1）, and the false discovery rate must be less than 0.05. Results When the mean temperature was 25.6 ℃, Chrysomyia Megacephala emerged 6 d after they pupated. A total of 43 408 pieces of Unigenes were obtained and their mean length was 905 bp, of which 32 500, 18 720, 13 542, 9 191 and 18 720 pieces were annotated by NR, SWISS-PORT, Pfam, STRING and KEGG databases. According to the discrepant gene analysis of pupae in two different growing periods, the number of genes with variants ranged from 801 to 5 307, and the total number of discrepant genes was 45 676. Conclusion The gene expressions of the transcriptome data of Chrysomyia Megacephala pupae in different growing periods are different. The results provided a good foundation for further research on the transcriptome changes in each period of the pupae of sarcosaprophagous flies and provided the basis for exploring the genes associated with the growth of Chrysomyia Megacephala pupae.

Research Progress on Species Identification of Sarcosaprophagous Flies / 法医学杂志

Estimation of postmortem interval （PMI） has always been one of the difficult problems for forensic scientists. It is especially hard to estimate the PMI of highly decomposed corpses in the wild or in secluded houses with conventional methods. Therefore, application of insect evidence at the scene is usually required for estimation. Sarcosaprophagous flies of different species have totally different developmental rates. In actual cases, direct measurement of the body length of the larvae, calculation of accumulated temperature and succession stages without species identification, or calculation based on incorrect species identification would often lead to a large deviation between the calculated results and the real PMI. This mistake would also mislead the case investigation. Therefore, accurate species identification should be implemented before any PMI estimation of decomposed corpses with forensic entomological methods. This article reviews the general and ultramicroscopic species identification and molecular biological species identification methods of different stages of sarcosaprophagous flies, in order to provide new ideas and methods for related research and practice, and provide reference for the application and promotion of forensic entomology in the front line of public security.

Progress and Application of Entomological Evidence in Forensic Science / 法医学杂志

Entomological evidence provides entry points and clues for cases detection, in terms of estimation of the postmortem interval （PMI）, and place and cause of death. In recent years, the feasibility of entomological evidence in practice has been proved by theories and cases. It especially plays an important role in the investigation of cases with unnatural death, no monitoring, and highly corrupt cadaver. However, there are still some key issues to be further studied and standardized before the application of entomological evidence to forensic practice, to improve the effect of entomological evidence in forensic investigation and trial. This paper retrospectively reviews key studies of the application of entomological evidence in forensic science, mainly including discussion of forensic entomology inspection standard, identification studies of sarcosaprophagous insect species, collection of sarcosaprophagous insect growth and succession data under different environments and forensic entomotoxicology. With the rapid development of information technology and biotechnology, applying artificial intelligence and whole genome sequencing technology in forensic entomology has become a new research direction, which can improve the application value and range of entomological evidence in forensic science.

Development of Dermestes Maculatus at a Constant Temperature and Its Larval Instar Determination / 法医学杂志

Objective To establish the basic data for estimating minimum postmortem interval （PMImin） of heavily decayed and skeletonized remains by studying the development of Dermestes maculatus DeGeer （Coleoptera: Dermestidae）. Methods The developmental stages of Dermestes maculatus were observed at four constant temperatures of 20 ℃, 24 ℃, 28 ℃ and 32 ℃, and the changes in body length were also examined as the biological indicator to estimate larval day-age and instar. Results The total developmental time from egg to adult at 20 ℃, 24 ℃, 28 ℃ and 32 ℃ were （126.7±10.6） d, （69.4±8.2） d, （50.4±8.4） d and （49.6±6.5） d, respectively. The body length increased gradually, but changed irregularly as a whole. Conclusion The study provides basic data on the development and growth of Dermestes maculatus, especially on its developmental duration as a significant value for estimating PMImin of heavily decayed and skeletonized remains. Nevertheless, the change of body length is not found to be the best biological indicator for instar determination.

Research Progress on Developmental Biology of Sarcosaprophagous Insects / 法医学杂志

Forensic entomology provides a feasible way to estimate postmortem interval (PMI), of which the growth and development of sarcosaprophagous insects is the most widely used indicator in forensic practice. Over the years, forensic entomologists have carried out a large number of studies on the development biology of sarcosaprophagous insects. This paper illustrates the main factors that affect the development of sarcosaprophagous insects, including temperature, humidity, light, food types and poisons. The development indicators of sarcosaprophagous insects were reviewed from the perspectives of morphology, differential gene expression and biochemical characteristics. It is emphasized that future research of development biology on sarcosaprophagous insects should fully absorb and integrate the methods of artificial intelligence and omics, and the research object also needs further expansion in order to establish a more objective and more accurate PMI estimation method.

Identification of Common Sarcosaprophagous Flies in the Yangtze River Delta by COⅠ Gene / 法医学杂志

OBJECTIVES@#To identify the common sarcosaprophagous flies in the Yangtze River Delta based on mitochondrial cytochrome c oxidase subunit Ⅰ(COⅠ) gene sequence and verify the reliability of this method.@*METHODS@#Seven common genetically stable sarcosaprophagous flies in three families and six genera were collected from large domestic pig carcasses placed in the field and cultured in the laboratory for many generations. The whole genome DNA was extracted and the COⅠ gene fragment was amplified. The forward and reverse sequencing was followed by splicing. The base composition of the amplified fragment and the rate of interspecific evolutionary divergence were analyzed by software such as Mega 7.0.26. The phylogenetic tree of COⅠ gene sequence of common necrophagous flies in the Yangtze River Delta was established by neighbor joining (NJ) method and unweighted pair-group method with arithmetic means (UPGMA) method.@*RESULTS@#The average base composition of different flies was A(30.14%), T(38.23%), C(15.98%), G(15.65%). The rate of interspecific evolutionary divergence ranged from 2.2% to 15.3%, the lowest rate was between Chrysomya megacephala and Chrysomya pinguis, the highest rate was between Muscina stabulans and Boettcherisca peregrina.@*CONCLUSIONS@#COⅠ gene can be used to identify the common necrophagous flies in the Yangtze River Delta.

Sucesión de entomofauna de interés criminalista en modelo experimental de Cavia porcellus / Succession of entomofauna of criminal interest in experimental model of Cavia porcellus

La Entomología Forense (EF) es una rama de las ciencias médico-legales (Ortloff et al. 2012), que utiliza como herramienta clave a insectos y otros artrópodos que interactúan con un cuerpo en descomposición (Gennard, 2007). El presente estudio se basó en un diseño observacional descriptivo de tipo poblacional, en el que se evaluó la diversidad de dípteros de la familia Calliphoridae y Muscidae en la parroquia rural de Posorja, Guayaquil. En el proceso experimental en los biomodelos A y B, se obtuvo que las temperaturas mínimas diarias presentaron un promedio de 21 °C (entre 20 °C y 22 °C) en lo que respecta a las temperaturas máximas diarias presentaron un promedio de 29,50 °C (entre 26 y 31 °C). En ambos biomodelos experimentales A y B, la exposición directa a los rayos solares y el espacio de liberación abierto contribuyo con el desarrollo de los cambios post mortem de descomposición cadavérica, como son el cromatismo, hinchado, licuefacción y reducción. A la par, de la ocurrencia de la sucesión de dípteros de interés criminalisticos, pertenecientes a las familias Calliphoridae y Muscidae. La importancia de de este acercamiento a la fauna de interés forense dentro de la provincia de Posorja, se muestra en el abanico de oportunidades que abre, con miras a la futura incorporación de la entomología forense en investigaciones criminalísticas en casos de homicidios y negligencias. Sin embargo, todavía eisten numerosas dificultades dificultades para este tipo de estudio en el Ecuador, por lo que se propone divulgar la escala estacional y espacial de de estas investigaciones, a través de convenios que permitan replicarlos en diversas partes del país en distintos ecosistemas y bajo distintas condiciones climáticas(AU)

Forensic Entomology (PE) is a branch of the medico-legal sciences (Ortloff et al. 2012), which uses insects and other arthropods that interact with a decomposing body as a key tool (Gennard, 2007). The present study was based on a descriptive observational design of a population type, in which the diversity of diptera of the Calliphoridae and Muscidae families in the rural parish of Posorja, Guayaquil was evaluated. In the experimental process in biomodels A and B, it was obtained that the minimum daily temperatures presented an average of 21 ° C (between 20 ° C and 22 ° C) with regard to the maximum daily temperatures presented an average of 29, 50 ° C (between 26 and 31 ° C). In both experimental biomodels A and B, direct exposure to sunlight and the open release space contributed to the development of post-mortem changes in cadaveric decomposition, such as chromaticism, swelling, liquefaction and reduction. At the same time, the occurrence of the succession of diptera of criminalistic interest, belonging to the Calliphoridae and Muscidae families. The importance of this approach to the fauna of forensic interest within the province of Posorja is shown in the range of opportunities that it opens up, with a view to the future incorporation of forensic entomology in criminal investigations in cases of homicides and negligence. However, there are still numerous difficulties for this type of study in Ecuador, so it is proposed to disclose the seasonal and spatial scale of these investigations, through agreements that allow replication in different parts of the country in different ecosystems and under different weather conditions(AU)

Diversity of sarcosaprophagous Diptera (Calliphoridae, Sarcophagidae) in organic and conventional mango plantations in the Brazilian semi-arid region

ABSTRACT Increase on intensive monocultures is an important threat to Caatinga biodiversity decrease. The use of chemical insecticides directly affects insect natural enemies of crops, but also non-target species. We aimed to test whether the diversity of sarcosaprophagous flies would be affected by agricultural management (conventional and organic). Adult dipterans were collected monthly, from July 2015 to January 2016, using a grid composed of 12 suspended traps distant 20 m from each other, baited with 150 g of sardine or bovine spleen, in each type of management. We first calculated α-diversity by Hill numbers, then evaluated the difference of diversity metrics by ANOVA. We also graphically visualise the species composition distribution by NMDS. We recorded a total of 4,651 specimens of 15 Calliphoridae and Sarcophagidae species. Chrysomya spp. were the dominant species (93.7% of all blowflies). Sarcophagidae species were sampled in low abundance, but with exclusive presence of some species in each treatment (e.g. B laesoxipha stallengi and Ravinia belforti in organic management). Although there was no difference on sarcosaprophagous fly diversity between treatments, we demonstrate empirically that mango plantations in the Caatinga harbour diverse and abundant assemblages of sarcosaprophagous dipterans, whose richness of species is comparable to other ecosystems such as rainforests and urban areas. Our results expand the known geographical range for forensically relevant species by providing a checklist of sarcosaprophagous dipterans that can thrive in agroecosystems in the Caatinga and, potentially, colonize cadavers disposed in that type of environment.

Application of Chrysomya rufifacies (Diptera, Calliphoridae) to Postmortem Interval Estimation in Southwest China: A Case Report / 法医学杂志

Insect samples found on human corpses can provide the information important to estimating the minimum postmortem interval (PMImin). A female cadaver, found in a deserted factory in Chongqing of China, was confirmed as a homicide case after the forensic investigation and autopsy. Determining the time of death was difficult due to the inconsistent degree of decomposition in different parts of the decedent. The insect specimens found on the cadaver were identified to be Chrysomya rufifacies (C. rufifacies, Macquart) by morphology and mitochondrial DNA sequence analysis. The PMImin was estimated to be 452 h, based on the developmental rate of C. rufifacies. The PMImin was estimated suc-cessfully to be almost precise, which provided an important entomological evidence for case investiga-tion and suspect prosecution. In so doing, this highlights the usefulness of entomological evidence of specific species in the geographic area for PMI accurate estimation, especially in the case of advanceddecomposed corpses.

Molecular phylogeny of Monomorium pharaonis (Hymenoptera: Formicidae) based on rRNA sequences of mitochondrial gene

Many studies on forensic entomology have focussed on Diptera and lack the Hymenopteras population. Nonetheless, hymenopterans are part of the entomofaunal colonization of a corpse. Morphologically, it is difficult to identify and distinguish between them. In this study, using mitochondrial DNA knowledge, the molecular analysis was performed to classify the recovered species of hymenoptera collected from rabbit carcass, quickly and accurately. A molecular identification method with a 251-bp fragment of the 16S ribosomal gene RNA (16S rRNA) from a single ant species was evaluated. The maximum likelihood method analysis has recovered a generally well-supported phylogeny, with most taxa and species groups currently being recognized as monophyletic. The aculeate consists of some Hymenoptera’s best known. Their sister group has traditionally been considered in Ichneumonoidea. In addition, Trigonaloidea was found as the aculeates’ sister group and Crabronidae in Apoidea forming the Formicidae’s sister group. These results will play an important role in the implementation of the Saudi database forensically relevant ants.

COⅠ and 16S rDNA Sequence Identification of Common Necrophagous Flies in Fujian Province / 法医学杂志

Objective To identify the species of common necrophagous flies in Fujian Province by gene fragment sequences of mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） and 16S ribosomal deoxyribonucleic acid （16S rDNA）, and to explore the identification efficacy of these two molecular markers. Methods In total 22 common necrophagous flies were collected from the death scenes in 9 different regions in Fujian Province and DNA was extracted from the flies after morphological identification. The gene fragments of COⅠ and 16S rDNA were amplified and sequenced. All the sequences were uploaded to GeneBank and BLAST and MEGA 10.0 software were used to perform sequence alignment, homology analysis and intraspecific and interspecific genetic distance analysis. The phylogenetic trees of DNA fragment sequences of COⅠ and 16S rDNA of common necrophagous flies in Fujian Province were established by unweighted pair-group method with arithmetic means （UPGMA）, respectively. Results The flies were classified into 6 species, 5 genera and 3 families by morphological identification. The results of gene sequence analysis showed that the average number of interspecific and intraspecific genetic distance of 16S rDNA ranged from 1.8% to 8.9% and 0.0% to 2.4%, respectively. The average number of interspecific and intraspecific genetic distance of COⅠ ranged from 7.2% to 13.6% and 0.0% to 6.3%, respectively. Conclusion The gene sequences of COⅠ and 16S rDNA can accurately identify the species of different necrophagous flies, and 16S rDNA showed higher value in species identification of common calliphoridae necrophagous flies in Fujian Province.

The carrion beetle Oxelytrum discicolle (Coleoptera: Silphidae) and the estimative of the minimum post-mortem interval in a forensic case in Brasília, Brazil

Abstract The order Coleoptera is considered second highest in forensic importance in criminal cases involving violent death, because members of this order increase in richness and abundance as the decomposition process progresses. We present here the first Forensic Entomology (FE) case in Brazil where the carrion beetle, Oxelytrum discicolle (Coleoptera: Silphidae), was used to estimate the minimum post-mortem interval (m-PMI) in a forensic case. A female corpse, at an advanced stage of decomposition, was found on a deserted highway near Brasília/Brazil. The corpse was removed from the site, and insect samples were collected on and inside the corpse at the Forensic Medicine Institute of the Federal Police, following the specific FE protocols. The m-PMI was estimated considering two techniques, the pre-appearance interval (PAI) and the accumulated degree-days (ADD). The development stage of the larvae of O. discicolle allowed investigators to propose that this species was part of an earlier colonization, soon after death, with total length and prothorax width compatible with third instar larvae. Adult females of O. discicolle oviposited on the corpse at least 20 days before it was removed from the site where it was found, thus characterizing the m-PMI. This is the first case in which information on the development of the O. discicolle was used in a criminal investigation in Brazil.