RESUMO
In forensic entomology, rapid and unambiguous identification of blowfly species is a critical prerequisite for accurately estimating the post-mortem interval (PMI). The conventional diagnosis of cadaveric entomofauna based on external characters is hampered by the morphological similarities between species, especially in immature stages. Genetic analysis has been shown to allow precise and reliable diagnosis and delimitation of insect species. Nevertheless, the taxonomy of some species remains unresolved. This study was focused on improving the effectiveness and accuracy of analysis based on the widely used cytochrome c oxidase subunit I barcode region (COI barcode, 658 bp), complemented by other mitochondrial and nuclear regions, such as cytochrome b (Cyt-b, 307 bp) and the second internal transcribed spacer (ITS2, 310-331 bp), for the identification of Southern European blowflies. We analyzed a total of 209 specimens, collected from 38 human corpses, belonging to three Calliphoridae genera and seven species: Chrysomya (Ch. albiceps), Calliphora (C. vicina and C. vomitoria), and Lucilia (L. sericata, L. ampullacea, L. caesar and L. illustris). These species are the most common PMI indicators in Portugal. The results revealed that unambiguous separation of species of the Lucilia genus requires different loci from the barcode region. Furthermore, we conclude that the ITS2 (310-331 bp) molecular marker is a promising diagnostic tool because its inter-specific discriminatory power enables unequivocal and consistent distinctions to be made, even between closely related species (L. caesar-L. illustris). This work also contributes new genetic data that may be of interest in performing species diagnosis for Southern European blowflies. Notably, to the best of our knowledge, we provide the first records of the Cyt-b (307 bp) locus for L. illustris and the ITS2 (310-331 bp) region for Iberian Peninsula Lucilia species.
Assuntos
DNA Mitocondrial/genética , DNA/genética , Dípteros/genética , Animais , Citocromos b/genética , Código de Barras de DNA Taxonômico , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Entomologia , Europa (Continente) , Comportamento Alimentar , Ciências Forenses , Haplótipos , Humanos , Reação em Cadeia da Polimerase , Mudanças Depois da Morte , Análise de Sequência de DNARESUMO
In Forensic Entomology the fast and accurate identification of insects collected at the scene of events is essential if errors are to be avoided in estimating infestation times of interest and determining the post-mortem interval (PMI). Traditional identification based on morphological characteristics can be complicated due to physical similarities between different species, especially at immature stages. Genetic analysis provides a fast and reliable identification method. In this paper, molecular analysis is focused on a fragment of 307bp of the mitochondrial DNA region that codes for cytochrome b (cyt b). Six species belonging three genera of Calliphoridae of forensic interest (Calliphora vicina, Calliphora vomitoria, Lucilia sericata, Lucilia caesar, Lucilia ampullacea, Chrysomya albiceps) were collected and identified. These are the most common species of cadaveric entomofauna on the Atlantic seaboard of the Iberian Peninsula. The results show the robustness of the cyt b locus as a diagnostic tool, since its nucleotide variability enables reliable distinctions to be drawn between species. This study also contributes new consense sequences which may be of interest for correct species identification.
Assuntos
Citocromos b/genética , DNA Mitocondrial/genética , Dípteros/genética , Animais , Cadáver , Entomologia , Comportamento Alimentar , Patologia Legal , Humanos , Reação em Cadeia da Polimerase , Mudanças Depois da Morte , Análise de SequênciaRESUMO
The study of X-chromosomal short tandem repeats (X-STRs) can complement the analysis of autosomal and Y-STRs. A decaplex system for the X-chromosome genetic markers, DXS8378, DXS9898, DXS7133, GATA31E08, GATA172D05, DXS7423, DXS6809, DXS7132, DXS9902 and DXS6789, was used to study a population sample of Santa Catarina, Brazil. 184 individuals (72 female and 112 male samples) were typed. DNA was amplified in a multiplex reaction and the automatic detection performed using capillary electrophoresis. Allele frequencies and some forensic parameters were calculated.
Assuntos
Cromossomos Humanos X , Frequência do Gene , Genética Populacional , Sequências de Repetição em Tandem , Brasil , Impressões Digitais de DNA , Eletroforese Capilar , Feminino , Marcadores Genéticos , Humanos , Masculino , Reação em Cadeia da PolimeraseRESUMO
The state of Santa Catarina (Brazil) is known to have represented a cultural crossroads in South America due to several historic migrations mainly from Europe and Africa. We set out to scrutinize whether the genetic imprint of these migrations could be traced through analysis of the matrilineal gene pool of the Catarinenses. The entire control region of the mitochondrial DNA was studied in 80 healthy and maternally unrelated individuals. The analysis of haplogroup distribution revealed that this population is extremely heterogeneous, showing the coexistence of matrilineal lineages with three different phylogeographic origins. European lineages are the most frequent due mainly to the impact of relatively recent migratory waves from Europe. In spite of this, Native American lineages and African lineages incorporated with the slave trade are also present in noticeable proportions. The strikingly high variability generated by intense gene flow is mirrored in a high sequence diversity (0.9930) and power of discrimination (0.9806). Thus, analysis of the entire mitochondrial DNA control region emerges as a valuable tool for forensic genetic purposes in this highly admixed population, an attribute common to several present-day Latin American populations.
Assuntos
DNA Mitocondrial/genética , Genética Populacional , Sequências de Repetição em Tandem , Brasil , Regiões Determinantes de Complementaridade/genética , Impressões Digitais de DNA , Emigração e Imigração , Europa (Continente) , Frequência do Gene , Genótipo , Haplótipos , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo Genético , Grupos Raciais/genéticaRESUMO
One-hundred and nine unrelated and healthy males from Santa Catarina, Brazil were included in this study. Allele frequencies and gene diversities for the loci DYS456, DYS458 and DYS448 were calculated. A comparison between our population and others was performed.
Assuntos
Cromossomos Humanos Y , Variação Genética , Genética Populacional , Haplótipos , Sequências de Repetição em Tandem , Brasil , Impressões Digitais de DNA , Frequência do Gene , Humanos , Masculino , Reação em Cadeia da PolimeraseRESUMO
Medico-legal entomology, one area in the broad field of entomology, is routinely used in forensic applications. Insects are often collected from a corpse during criminal information related to the body, but requires the fast and accurate identification of the species attracted to the remains. The local entomofauna in most cases is important for explaining entomological evidence. The survey of the local entomofauna has become a fundamental first step in forensic entomological studies, because different geographical distributions, seasonal and environmental factors may influence the decomposition process and the occurrence of different species on corpses. A morphological and DNA-based methods for species identification were used in this study. Thirty-two cases are reported from indoors and outdoors conditions. Specimens were collected from corpses during autopsy procedures in the National Institute of Legal Medicine, Portugal, and cases were summarized by sex, death local, month of discovery, probable cause of death, species found and number of analyzed specimens. Just eight species, mainly Calliphoridae together with one Sarcophagidae were reported from corpses. The DNA sequencing was performed to study the mitochondrial encoded subunit I of the cytochrome oxidase gene. The aim of this work was the beginning of a database of the cadaveric entomofauna in Portugal.
Assuntos
DNA Mitocondrial/genética , Dípteros/genética , Animais , Complexo IV da Cadeia de Transporte de Elétrons/genética , Entomologia , Meio Ambiente , Comportamento Alimentar , Feminino , Antropologia Forense , Humanos , Masculino , Reação em Cadeia da Polimerase , Portugal , Mudanças Depois da Morte , Análise de Sequência de DNAAssuntos
Frequência do Gene/genética , Genética Médica , Repetições de Microssatélites/genética , Grupos Populacionais/genética , Pareamento de Bases/genética , Mapeamento Cromossômico , DNA/genética , Humanos , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase/métodos , PortugalRESUMO
POPULATION: One hundred unrelated females and 100 unrelated males, autochthonous, healthy, from the North of Portugal.
Assuntos
Cromossomos Humanos X , Genética Populacional , Sequências de Repetição em Tandem , Impressões Digitais de DNA , Feminino , Frequência do Gene , Humanos , Masculino , Reação em Cadeia da Polimerase , PortugalRESUMO
POPULATION: A total of 184 healthy unrelated individuals (70 females and 114 males), autochthonous from Santa Catarina, Brazil.
