Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.
Aedes/*parasitology ; Animals ; Anopheles/*parasitology ; Culex/*parasitology ; Dirofilaria immitis/genetics/*isolation & purification ; Dirofilaria repens/genetics/*isolation & purification ; Egypt ; Entomology/methods ; Female ; Multiplex Polymerase Chain Reaction/*methods ; Parasitology/methods ; Sensitivity and Specificity ; Wuchereria bancrofti/genetics/*isolation & purification

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.
Aedes/*parasitology ; Animals ; Anopheles/*parasitology ; Culex/*parasitology ; Dirofilaria immitis/genetics/*isolation & purification ; Dirofilaria repens/genetics/*isolation & purification ; Egypt ; Entomology/methods ; Female ; Multiplex Polymerase Chain Reaction/*methods ; Parasitology/methods ; Sensitivity and Specificity ; Wuchereria bancrofti/genetics/*isolation & purification

Forensic entomotoxicology is a branch of forensic medicine, which applies entomology, toxicology and other related studies to solve the poisoning cases. It has an obvious advantage in the investigation on poisoning death. Based on the expounding definition and research of entomotoxicology, this paper reviews research progress and application value in some aspects of forensic medicine, such as the effects of drugs/toxins on the growth and development of sarcosaphagous insects and the qualitative and quantitative analysis of the drugs/toxins in the poisoned body tissue.
Animals ; Death ; Entomology/methods* ; Forensic Medicine/methods* ; Humans ; Insecta ; Postmortem Changes

OBJECTIVE: To document the arthropod succession pattern and to identify forensically important species in northeastern Egypt (32° 15' E and 30° 36' N) for the first time. METHODS: Carcasses were exposed in an open area for 60 days during summer season. Ambient daily temperature (maximum and minimum) and relative humidity (RH) were recorded and existing keys were used for identification of different species. RESULTS: During the period of study, the mean of maximum and minimum temperatures were 34.85 °C and 29.2 °C respectively, while the mean of RH was 53.5%. Four stages of decomposition were observed: fresh, bloat, decay and dry. The most abundant orders were found to be Diptera, Coleoptera and Hymenoptera. Arthropods were collected belonging to 4 families of Diptera: Muscidae, Fanniidae, Calliphoridae and Sarcophagidae. While there were 2 families of Coleoptera: Dermestidae and Histeridae. Monomorium species was the only Hymenoptera family in this study. CONCLUSION The present work provided a basis for further studies dealing with insect colonization of carcasses in different seasons and locations in Egypt.
Animals ; Arthropods ; classification ; physiology ; Coleoptera ; Diptera ; Egypt ; Entomology ; Feeding Behavior ; Forensic Medicine ; methods ; Hymenoptera ; Insecta ; classification ; Rabbits ; Rats ; Seasons ; Temperature

Estimating postmortem interval (PMI) is always the emphasis and difficulty in forensic practice. Forensic entomology plays a significant indispensable role. Recently, the theories and technologies of forensic entomology are increasingly rich. But many problems remain in the research and practice. With proposing the Daubert standard, the reliability and accuracy of estimation PMI by forensic entomology need more demands. This review summarizes the application of the Daubert standard in several aspects of ecology, quantitative genetics, population genetics, molecular biology, and microbiology in the practice of forensic entomology. It builds a bridge for basic research and forensic practice to provide higher accuracy for estimating postmortem interval by forensic entomology.
Animals ; Ecology ; Entomology/methods* ; Forensic Sciences/methods* ; Genetics, Population ; Insecta ; Molecular Biology ; Postmortem Changes ; Reproducibility of Results ; Time Factors

Diptera Calliphoridae is the first major kind of flies that appears on the decomposed corpses. In forensic entomology, according to the living characteristics of Calliphoridae flies, we could accurately estimate postmortem interval (PMI) in a murder or unidentified case and could provide useful clues to solve the case. This paper introduces the characteristics of the biology and morphology of Diptera Calliphoridae, and reviews the combined application of forensic entomology, molecular biology, mathematical morphology and toxicology.
Animals ; Autopsy ; Cadaver ; Diptera ; Entomology ; Forensic Anthropology ; Forensic Medicine/methods* ; Forensic Sciences ; Humans ; Postmortem Changes ; Time Factors

OBJECTIVE: To identify the common Sarcophagidae with a 278 bp fragment of cytochrome oxidase I in mitochondrial DNA and to obtain an unambiguous and rapid identification method for Sarcophagidae in forensic investigations. METHODS: Nineteen Sarcosaprophagous flies were collected from 16 locations in 12 Chinese provinces. All specimens were comprised of 4 species. The mtDNA of flies was extracted with SDSPK extraction method. Polymerase chain reaction was conducted in an Eppendorf 5331 thermal cycler. The PCR products were purified and sequenced and the obtained sequences were uploaded to GenBank. A neighbor-joining tree was constructed with MEGA4.0 package. RESULTS: The 19 Sarcosaprophagous flies were well clustered. The intraspecific variation within species varied from 0% to 3%, while the interspecific variations between species varied from 8% to 12%. CONCLUSION Congeneric species can be separated by the short fragment (278 bp region in the cytochrome oxidase subunit I gene), which will be instrumental for implementation of the Chinese Sarcophagidae database and lay a foundation for post mortem interval estimation in future forensic cases.
Animals ; DNA, Mitochondrial ; genetics ; Electron Transport Complex IV ; genetics ; Entomology ; methods ; Forensic Pathology ; methods ; Polymerase Chain Reaction ; Postmortem Changes ; Sarcophagidae ; classification ; enzymology ; genetics ; Species Specificity

OBJECTIVE: To explore the application of a 289bp fragment of the 16S rDNA gene to identify various species of sarcosaphagous Calliphorid flies. METHODS: Twenty-six Calliphorid flies were collected from 14 Chinese provinces. All specimens were properly assigned into three genera and six species. The DNA of the pectoralis was extracted using CTAB method. Then PCR amplification was done for the 289 bp fragment of the 16S rDNA gene. The PCR products were then purified and sequenced, and the obtained sequences were uploaded to GenBank. The phylogenetic tree was built by the neighbor-joining method and intraspecific and interspecific divergences were calculated by sequence analysis. RESULTS: The above 26 sarcosaphagous flies could be well clustered according to different genera and species. The evolutional intraspecific values were all zero, the evolutional interspecific variations varied from 0.3% to 6.5%. CONCLUSION The 289 bp fragment of the 16S rDNA of sarcosaphagous flies can be effectively used to identify most of the flies at species level. This method appears to be fast and low dissipative, which might be used to estimate postmortem interval by sarcosaphagous flies.
Animals ; DNA Primers ; DNA, Mitochondrial/genetics* ; DNA, Ribosomal/genetics* ; Diptera/genetics* ; Entomology ; Forensic Medicine/methods* ; Phylogeny ; Polymerase Chain Reaction/methods* ; RNA, Ribosomal, 16S/genetics* ; Rabbits ; Sequence Analysis, DNA ; Species Specificity

Species identification of sarcosaphagous insects is one of the important steps in forensic research based on the knowledge of entomology. Recent studies reveal that the application of molecular biology, especially the mtDNA sequences analysis, works well in the species identification of sarcosaphagous insects. The molecular biology characteristics, structures, polymorphism of mtDNA of sarcosaphagous insects, and the recent studies in species identification of sarcosaphagous insects are reviewed in this article.
Amino Acid Sequence ; Animals ; Base Sequence ; DNA, Mitochondrial/genetics* ; Diptera/genetics* ; Electron Transport Complex IV/genetics* ; Entomology ; Forensic Medicine/methods* ; Genes, Mitochondrial/genetics* ; Insecta/genetics* ; Polymerase Chain Reaction ; Polymorphism, Genetic ; RNA, Ribosomal/genetics* ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVE: To compare effects of three different methods for mtDNA extraction from common sarcosaphagous insects including cetyl trimethyl ammonium bromide (CTAB) method, sodium dodecyl sulfate-potassium acetate (SDS-KAc) method and sodium dodecyl sulfate-proteinase K (SDS-PK) method. METHODS: Seventy-two insects from four species [Chrysomya megacephala (Fabricius, 1784), Eusilpha bicolor (Fairmaire, 1896), Paraeutrichopus pecoudi (Mateu, 1954), Vespa velutina (Lepeletier, 1836)] were collected from the corpses of the rabbits in Changsha district. The total DNA of above samples was extracted by CTAB, SDS-Kac and SDS-PK methods. The purity and concentration of DNA were examined by protein-nucleic acid spectrophotometry, and mtDNA were amplified by specific primers and PCR products were detected by agarose gel electrophoresis. Then PCR products were sequenced and subsequently up-loaded to GenBank. RESULTS: mtDNA was successfully extracted with three methods from most of the samples. The SDS-PK method was better in DNA purity compared to other methods and the CTAB method was superior in extracting DNA from old samples, while SDS-KAc method showed no significant difference for extraction effects of different samples. CONCLUSION The most appropriate method should be chosen depending on different situations. SDS-PK method is expected to obtain high-quality DNA, while CTAB method is preferred in extracting obsolete samples. SDS-KAc method is low cost and can be used in various kinds of preliminary experiments.
Animals ; Coleoptera/genetics* ; DNA Primers ; DNA, Mitochondrial/isolation & purification* ; Diptera/genetics* ; Electrophoresis, Agar Gel ; Entomology ; Forensic Medicine/methods* ; Gene Amplification ; Insecta/genetics* ; Polymerase Chain Reaction/methods* ; Quaternary Ammonium Compounds/chemistry* ; Rabbits ; Reproducibility of Results ; Sequence Analysis, DNA ; Sodium Dodecyl Sulfate/chemistry*