Background: Pomacea canaliculata (Lamarck, 1819) is an invasive freshwater snail in the Philippines that damages crops but is consumed as food. It is known to harbor parasites, some of which are pathogenic to humans. Objectives: The objective of this study is to examine P. canaliculata individuals present in Metro Manila for parasite infection and identify the parasites and other organisms associated with the snail using molecular identification. Methodology: P. canaliculata were collected from rivers and marketplaces in Metro Manila. Individuals were crushed and digested in Ash's digestive fluid and observed under a microscope. Collected parasites were subjected to DNA barcoding of the COI gene for putative identification. Results: A total of 462 snails were gathered from 15 sites, eight of which were market areas that sourced the snails from outside Metro Manila. No known parasites were found. Two snails were found to contain insect larvae in the mantle (0.43% infection). The closest BLAST matches for the two insect larvae were the chironomid fly Nilodorum tainanus (91.0% identity) from a snail in Sucat, Muntinlupa, and another chironomid Parachironomus sp. (92.8% identity) found in a snail originally from Cavite and brought to Calumpang, Marikina. Conclusion This study is the first report of the presence of chironomids in Philippine P. canaliculata. This could have an impact on the allergenic status of these mollusks if consumed while containing these chironomids. The absence of infection of other medically important parasites is possibly due to the patchy distribution of the snails and few interactions with the definitive hosts of known parasites.
Chironomidae ; Diptera

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.
Animals ; Cadaver ; Diptera/genetics* ; Phylogeny ; Reproducibility of Results ; Rivers

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.
Animals ; Artificial Intelligence ; Developmental Biology ; Diptera ; Entomology ; Insecta ; Postmortem Changes

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.
Animals ; Artificial Intelligence ; Diptera ; Entomology ; Forensic Sciences ; Postmortem Changes ; Retrospective Studies

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.
Animals ; Autopsy ; Cadaver ; Diptera ; Entomology ; Larva ; Postmortem Changes

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.
Animals ; Calliphoridae ; Diptera ; Female ; Male ; Reproducibility of Results

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.
Animals ; Autopsy ; Diptera ; Entomology ; Insecta ; Larva ; Postmortem Changes

Aims: The primary aim of the present study is to evaluate the effect of rearing substrates on the nutritional content of black soldier fly larvae (BSFL) by incorporating Cupriavidus necator cells containing intracellular polyhydroxybutyrate (PHB) in BSFL diet to further increase the protein content and simultaneously to biologically extract the polymer by utilizing the digestive system of BSFL. The potential application of BSFL as a biological PHB extraction agent was determined. Methodology and results: Two feeding strategies consists of a mixture of protein (P) to carbohydrate (C) with a ratio of P50:C50 food waste (control feeding) and feed with bacterial cells (modified feeding). A comparison on the proximate analysis between this research and two commercially available products were conducted. Feeding BSFL with P50:C50 food waste revealed the highest crude protein content of 81.3 ± 0.2%. Additional bacteria cells in the BSFL diet, however, showed a negligible decrease in crude protein content of 0.67% as compared to the control feeding. Howbeit, this results comparably higher in contrast to the commercial products, with increment of crude protein content by 12.1% and 40.8%, respectively. Conclusion, significance and impact of study Two desirable products were obtained from the feeding with cells: (1) high protein content of BSFL and (2) biologically extracted polymer. This is the first study to demonstrate the utilization of BSFL as a biological extraction agent to partially extract biopolymer and increase the protein content by feeding with cells.
Diptera ; Polyhydroxybutyrates ; Animal Feed

Leaf miner is one of the major pests on safflower, which causes yield loss and poor quality seriously. "Weihonghua", "nine safflower varieties" and "three chemical insecticides" as materials that used to evaluate variety and regularity of leaf miner, safflower resistant level, and different proportions insecticides in field efficiency test. The results showed that Liriomyza sativae and L. huidobrensis accounted for 80%, the peak period of two pests was all in July; but Phytomyza horticola is relative less, its peak period occured in June. Three were great difference of resistance to leaf miner among safflower varieties, FQ12 and YJ65 expressed higher resistibility to leaf miner by ratio method. With abamectin 2% emulsifiable concentrate diluted for 2 000 times, or the mixture three insecticides(bifenthrin 20% water emulsions, thiamethoxam 25% water dispersible granule, abamectin 2% emulsifiable concentrate=1∶1∶1) diluted for 3 000 times, which were sprayed on leaves at squaring stage and lethal rate was 96% after 48 h in the study. Through comparative study on the variety and regularity of leaf miner, screen for resistant varieties to leaf miner and for high efficiency pesticide. The study provides theoretical basis and reference for integrated pest management of leaf miner.
Animals ; Carthamus tinctorius ; Diptera ; Insecticides ; Pesticides ; Thiamethoxam

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.
Animals ; DNA, Ribosomal/genetics* ; Diptera/genetics* ; Humans ; Phylogeny ; RNA, Ribosomal, 16S/genetics* ; Sequence Analysis, DNA ; Species Specificity