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.

Potential application of Gustatory Receptor 1 (CmegGr1) gene as a molecular marker for identification of Chrysomya megacephala (Diptera: Calliphoridae)

@#Chrysomya megacephala larvae can easily be identified using cheap traditional microscopy techniques. Nevertheless, identification using taxonomy keys may be hampered, if the morphological characteristics of the larvae are incomplete, or immature for microscopic identification. To overcome the difficulty of species determination, molecular identification has gained relevance and is applied in forensic investigations. This study aimed to identify a novel target gene, known as the gustatory receptor 1 gene (CmegGr1), which has never been used for identification. The third instar larvae of Ch. megacephala (n = 30) and eight other forensically important fly species were obtained from two sources; rabbit carcasses and the Forensic Entomology Unit collection. Their DNAs were extracted and the CmegGr1 gene was amplified using polymerase chain reaction (PCR). The resulting sequences were subjected to phylogenetic analysis. A 209 bp fragment of the CmegGr1 gene was successfully amplified in 80% (24/30) of Ch. megacephala samples, while all of the non-Ch. megacephala species were not amplified. The phylogenetic analysis revealed that the evolutionary tree of CmegGr1 shares many traits with the 21a gustatory receptors of Calliphora stygia and Lucilia cuprina (Gr21a), which are also classified as necrophagous fly species. The high specificity of species identification was demonstrated in the present study using DNA barcoding, which led to the conclusion that the CmegGr1 gene could serve as an alternative marker for identifying Ch. megacephala.

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.

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.

Epidemiological determinants of Chrysomya (Diptera: calliphoridae) infestation in layer farms of Minas Gerais, Brazil / Determinantes epidemiológicos da infestação por Chrysomya (Diptera: Calliphoridae) em granjas avícolas de postura de Minas Gerais, Brasil

Aspects related to the epidemiology of Diptera belonging to genus Chrysomya were studied in order to determine predisposing factors for their occurrence in commercial laying hens farms from the state of Minas Gerais, Brazil. An observational, analytical and sectional survey was conducted to verify the main epidemiological determinants of infestations by Chrysomya spp. Forty-three farms were visited in 2012 and these 13 (30,23%) were infested by Chrysomya putoria, 4 (9,30%) by Chrysomya megacephala and 2 (4,65%) by Chrysomya albiceps and in 2 farms (4,65%) was not possible to identify the species of Chrysomya. The epidemiological determinants for the occurrence of infestations by Chrysomya spp. were investigated using multiple logistic regression models. Housing clustering type and its conditions were important factors involved in epidemiology of these species (OR=5.05, 95% CI = 2.66 to 9.58, p = 0.000). It was also observed that the increase of the humidity of manure leads to increase chances of infestations (OR = 1.81, 95% CI = 1.36 to 2.40, p = 0.000). The presence of the beetle "lesser mealworm" (Alphitobiusdiaperinus) (OR = 0.15, 95% CI = 0.067 to 0.36, p = 0.000) was characterized as a protective factor against infestations, probably due to predation behavior of the beetle. Some regions of the state were characterized as lower risk areas for Chrysomya spp. infestation as compared to other regions. Management measures, such as constant monitoring of water leaks on manure, adequate composting of dead poultry and periodic removal of manure could assist in decreasing the presence of Chrysomya spp. inside the laying houses.

Aspectos relacionados à epidemiologia das infestações por dípteros do gênero Chrysomyaforam estudados com o objetivo de determinar os fatores predisponentes para sua ocorrência em granjas comerciais de postura do estado de Minas Gerais, Brasil. Foi realizado um estudo observacional, analítico e seccional para verificar os principais determinantes epidemiológicos das infestações por Chrysomya spp. Foram visitadas 43 granjas avícolas em 2012 e dessas 13 (30,23%) apresentaram infestação por Chrysomya putoria, 4 (9,30%) por Chrysomya megacephala, 2 (4,65%) por Chrysomya albiceps e em 2 granjas (4,65%) não foi possível identificar as espécies de Chrysomya. A verificação dos determinantes epidemiológicos foi realizada utilizando modelos de regressão logística múltipla. Diferenças entre os núcleos das granjas avícolas e que não foram avaliadas neste estudo, apresentaram-se importantes na epidemiologia destas moscas (OR=5.05 95% IC = 2.66 a 9.58, p = 0.000). Observou-se também que o aumento na umidade do esterco dos aviários aumenta as chances de ocorrência de infestações (OR = 1.81, 95% IC = 1.36 a 2.40, p = 0.000). A presença de coleópteros popularmente conhecidos como "Cascudinhos", Alphitobiusdiaperinus (OR = 0.15, 95% IC = 0.067 a 0.36, p = 0.000), foi caracterizada como um fator protetor contra infestações, provavelmente devido ao comportamento predador deste coleóptero. Algumas regiões do estado apresentaram menores chances de ocorrência de infestações por Chrysomya spp. quando comparadas a outras regiões. Algumas medidas de manejo, como monitoramento constante de vazamentos de água no esterco, compostagem adequada de aves mortas e remoção periódica do esterco, poderiam auxiliar na diminuição da presença de Chrysomya spp. no interior dos galpões de postura.

Development of Chrysomya albiceps Wiedemann, 1819 (Diptera: calliphoridae) and Musca domestica Linnaeus, 1758 (Diptera: muscidae) reared together in pig tissues

Chysomya albiceps and Musca domestica are important for forensic entomology, and human and animal health. This study analyzed the effects of the coexistence of C. albiceps and M. domestica reared in four different assays in two pig tissues, brain and intestine: assay 1, interaction between the larvae of the same age; assay 2, interaction between larvae of C. albiceps 24 hours older than larvae of M. domestica; assay 3, interaction between larvae of M. domestica 24 hours older than the larvae of C. albiceps; assay 4, larvae of both species were reared together in flasks with a small supply of food. Weight of larvae, growth time and imago emergence frequency were studied. C. albiceps responded better than M. domestica under most conditions tested. Larvae of C. albiceps responded better in mixed cultures (together with M. domestica) than in pure cultures (larvae of the same species). In contrast, M. domestica responded better when reared in pure cultures. Both species presented shorter growth times when their larvae were reared in intestine tissue with larvae 24 hours younger than the larvae of the concurrent species, compared to their respective growth times in pure cultures. The results confirmed that trophic interactions are relevant to the successful colonization of carrion by C. albiceps. Coexistence of the two species may result in changes in values of their biological components. The results also help to shed light on the biology of the two species in carrion

Synanthropic diptera affecting layer poultry farms: a review / Dípteros sinantrópicos de importância para a avicultura de postura: uma revisão

The poultry farm of posture is an economic activity of great relevance to Brazil. Health aspects of flocks of laying chickens, such as the occurrence of infestations by parasites and poultry pests, influence significantly the productivity indicators. In this context, the control of synanthropic diptera is one of the challenges of the poultry farmers and professionals of this area. In Brazil, the control of flies in poultry environments is based mainly on the use of pesticides, while other alternatives are less frequent. Among the flies' species most regularly found in poultry farms are the Musca domestica, Chrysomya spp., Fannia spp., and others. This review aims at compiling the literature on the occurrence, impact on poultry systems, biology, epidemiology and control of the species of synanthropic flies considered important for the Brazilian poultry industry.(AU)

A avicultura de postura é uma atividade econômica de grande relevância para o Brasil. Aspectos sanitários dos plantéis de galinhas poedeiras, tais como infestações por parasitos e pragas avícolas, influenciam significativamente os indicadores de produtividade desse setor. Nesse contexto, o controle de dípteros sinantrópicos constitui um dos desafios de avicultores e profissionais da área. No Brasil, o controle de moscas em ambientes avícolas é baseado, sobretudo, no uso de pesticidas, ao passo que o uso de outras alternativas é menos recorrente. Entre as espécies de moscas mais frequentes em granjas avícolas de postura, destacam-se Musca domestica, Chrysomya spp., Fannia spp., entre outras. O objetivo desta revisão é realizar a compilação da literatura existente sobre a ocorrência, o impacto nos sistemas avícolas, a biologia, a epidemiologia e o controle das espécies de dípteros sinantrópicos consideradas importantes para a avicultura de postura brasileira.(AU)

Traumatic Myiasis of the Scalp in a 3-year-old Patient- A Case of Neglected Health Care

@#Traumatic myiasis, the invasion of live tissues by fly larvae, is a commonly observed phenomenon in animals like sheep and cattle, but is also witnessed sometimes among humans. It is a parasitic infestation in which the dipteran larvae invade the neglected open wounds and foul smelling body apertures of vertebrate animals thereby using the living or necrotic tissue of their host as a nutritional source. A case of severe human traumatic myiasis is reported from India in the scalp wound of a 3-year-old female patient. The causative larvae were identified as the third instars of Old World Screwworm Fly- Chrysomya bezziana (Diptera: Calliphoridae). The myiasis in the present case was a complication of an open neglected wound. Ignorance played a key role in non-treatment of the patient for a long period of time. The authors emphasize the necessity to raise awareness among the general public about the problem.

Forensically Important Blow Flies Chrysomya pinguis, C. villeneuvi, and Lucilia porphyrina (Diptera: Calliphoridae) in a Case of Human Remains in Thailand

This is the first study to report Chrysomya pinguis (Walker) and Lucilia porphyrina (Walker) (Diptera: Calliphoridae) as forensically important blow fly species from human cadavers in Thailand, in addition to Chrysomya villeneuvi (Patton) already known in Thailand. In 2016, a fully decomposed body of an unknown adult male was discovered in a high mountainous forest during winter in Chiang Mai province. The remains were infested heavily with thousands of blow fly larvae feeding simultaneously on them. Morphological identification of adults reared from the larvae, and molecular analysis based on sequencing of 1,247 bp partial mitochondrial cytochrome c oxidase subunit 1 gene (CO1) of the larvae and puparia, confirmed the above mentioned 3 species. The approving forensic fly evidence by molecular approach was described for the first time in Thailand. Moreover, neighbor-joining phylogenetic analysis of the CO1 was performed to compare the relatedness of the species, thereby affirming the accuracy of identification. As species of entomofauna varies among cases in different geographic and climatic circumstances, C. pinguis and L. porphyrina were added to the list of Thai forensic entomology caseworks, including colonizers of human remains in open, high mountainous areas during winter. Further research should focus on these 3 species, for which no developmental data are currently available.

Use of maggot therapy for treating a diabetic foot ulcer colonized by multidrug resistant bacteria in Brazil.

This study reports the efficacy of maggot therapy in the treatment of diabetic foot ulcer infected with multidrug resistant microorganisms. A 74 year old female patient with diabetes for over 30 years, was treated with maggot therapy using larvae of Chrysomya megacephala. The microbiological samples were collected to evaluate aetiology of the infection. The therapy done for 43 days resulted in a reduction of necrosis and the ulcer’s retraction of 0.7 cm2 in area. Analysis of the bacteriological swabs revealed the presence of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Further studies need to be done to confirm the role of maggot therapy in wound healing using a large sample and a proper study design.

Influence of Fluctuated Room Conditions on the Development of the Forensically Important Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae).

Aims: To describe the influence of the seasonal variations on the development of blow fly Ch. albiceps (Wiedemann) stages as regards to the fluctuated temperature, Study Design: Eggs and larvae of the Ch. albiceps were collected from rabbit carcasses, the development period for immature and adult stages under naturally circulating room conditions were observed. Place and Duration of Study: Observations were carried out for a year (June, 2012 – May, 2013) at Zoology Dept., Zagazig University, Zagazig, Egypt. Methodology: Groups of newly formed pupae were kept in rearing jars for adult emergence. The development time (in days) from emergence to egg laying and immature growth was determined. Results: The overall development time of the Ch. albiceps exhibited significant seasonal variations corresponding to temperature changes. High temperatures accelerate the overall development, whereas low temperatures slow it down. The average period between emerging of Ch. albiceps adults, mating, spawning, larval and pupal development at low temperatures in winter was sustained the record average of 25-51 days when reared at 11°C. This period, was decreased gradually to reach average of 19-27 days at 15°C in autumn, 16-23 days at 18°C in spring and 17- 19 days, in summer months when temperature average was 23°C. Adult emergence of the Ch. albiceps fly was positively affected by temperature, it ranged from 96.12±0.24%, in summer (23°C), to 81.79±0.77 in winter (11°C). Generally female emergence was in high rates more than males and the higher female ratio was during winter (1.62). Also changes in temperatures due seasonal variations showed significant effect on the pre-oviposition and incubation periods, female fecundity, percentage of egg hatching and time of larval development. Conclusion: Changes in temperature according to seasonal variation considered an important factor for the development time for the forensically important Ch. albiceps.

Screw-worm Myiasis of Prolapsed Rectum.

Background: Wound myiasis in the Indian subcontinent is most commonly caused by old world screw-worm (Chrysomya bezziana). Case Report and management: A 4-yearold malnourished girl presented with full thickness rectal prolapse following acute diarrhea with a large wound and screwworm myiasis of the rectum. Turpentine oil was applied to immobilize the maggots followed by manual extraction. Prolapse was successfully treated by manual reduction followed by strapping of the buttocks. Outcome: Child was thriving well and gained 2 kg weight in follow up after two weeks. Message: Parents should be educated about taking care of prolapsed rectum.

Oral Myiasis: Case report.

Oral myiasis is a rare disease caused by larvae of dipteran flies. Houseflies are strongly suspected of transmitting at least 65 diseases to humans, including typhoid fever, dysentery and cholera. Flies regurgitate and excrete wherever they come to rest and thereby mechanically are the root cause for disease organisms. A case of oral myiasis caused by Chrysomya bezziana in the maxillary anterior region in a 40‑year‑old patient is presented. Manual removal of maggots, and surgical debridement of wound was done followed by broad‑spectrum anti‑parasitic medications. A note on the identification of the larva and histopathology of the tissue is also highlighted here.

Oral myiasis – a case report

Oral myiasis is a rare disease caused by larvae of certain dipteran flies. A case of oral myiasis caused by chrysomya bezziana, in the maxillary anterior region in a 20 year old mentally challenged female patient is reported. Manual removal of the larvae by topical application of turpentine oil, surgical debridement of the oral wound, followed by oral therapy with broad spectrum antibiotics were used to manage the patient.

Sensitivity of last larval stadium of Chrysomya megacephala (Fabricius) to anti-allatin ageratochromene Precocene II.

Treatment of early and late third instar larvae of economically important insect C. megacephala with the ageratochromene precocene II resulted into precocious metamorphosis. The other effects included mortality, production of pupal-adult mosaics, reduction in percentage of adult emergence and formation of non-viable deformed adults that died after a few hours of emergence. The application of precocene II adversely hampered the normal growth, development and metamorphosis of the insect and the results show that precocene II has vast potential for excellent control of this insect of medico-veterinary importance and can be used in insect pest management programs.

Epidemiological study of myiases in the Hospital do Andaraí, Rio de Janeiro, including reference to an exotic etiological agent

Myiases are infestations with dipteran larvae in both necrosed and living tissues, the food source of these insects. These illnesses occur in warm humid climates, and are most frequent in developing countries. We assessed the epidemiological aspects and the influence of climate on the occurrence of myiases and the bioagents in patients admitted to the federal Hospital do Andaraí in Rio de Janeiro from February 2007 to 2008. The influence of abiotic factors (temperature, humidity, and rainfall) on the incidence of myiases was investigated by using the Pearson's correlation test. Of the 40 patients studied, the prevalence of myiases was higher in adults, particularly in the 40 to 65 year-old (37.5 percent) African descent males (57.5 percent). Most of the injuries were caused by trauma (62.5 percent). Some patients made use of licit (50 percent) and illicit drugs (17.2 percent). The occurrence of myiases was not affected by the abiotic factors investigated. The cases reported here were treated in only one hospital, indicating that the disease is relatively common. The most frequent bioagent was Cochliomyia hominivorax (Coquerel), but Cochliomyia macellaria (Fabricius), Chrysomya albiceps (Wiedemann) and Dermatobia hominis (Wiedemann) (Diptera: Calliphoridae) were also detected causing myiases. Chrysomya albiceps is an exotic etiologic agent of myiases.

Oral myiasis caused by chrysomya bezziana: A case report.

Myiasis is caused by members of the Diptera fly family that lay eggs on food, necrotic tissue, open wounds, and unbroken skin or mucosa. Although myiasis is primarily a problem of cattle and livestock in tropics, human infestation is uncommon and occurs accidentally in persons at extremes of age, who are debilitated or not able to ensure basic hygiene and wound cleanliness. The oral cavity is an uncommon site of infestation as was seen in the case reported here.

Effects of Temperatures on Larval Development of Chrysomya megacephala (Fabricius) and Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae): Application in Forensic Science / Jurnal Sains Kesihatan Malaysia

Chrysomya megacephala (Fabricius) and Chrysomya rufifacies (Macquart) are two important species of blow flies that could be utilised as indicators to estimate the Post Mortem Interval (PMI) of corpses in forensic science. Determination of PMI of corpses is based on the larva size and its development stages. Development studies of eggs, larvae and pupae of Ch. megacephala and Ch. rufifacies at 27ºC, 30ºC and 33ºC were carried out using an insect growth chamber. Data from these studies were used to construct growth curves and Accumulated Degree Hours (ADH) for both species. Ch. megacephala needs 9.15 days to complete one life cycle at 27ºC, 8.54 days at 30ºC and 6.76 days at 33ºC. Development for Ch. rufifacies is slower than Ch. megacephala and takes 9.92 day at 27ºC, 9.13 days at 30ºC and 7.44 days at 33ºC. Eggs of the two species hatch faster at 33ºC compared to the two other temperatures. Low value of ADH shows shorter life cycle of that particular species. This study’s findings are useful in estimating PMI of the corpses found under various environmental temperatures.

Nim (Azadirachta indica): uma alternativa no controle de moscas na pecuária / Neem (Azadirachta indica): an alternative for controlling flies associated with animal breeding

As moscas Lucilia cuprina, Chrysomya megacephala, Cochliomyia hominivorax e Musca domestica, estão entre as mais importantes pragas da pecuária causando miíases e como portadoras de microrganismos patogênicos. O propósito deste estudo foi testar a eficácia de várias concentrações do óleo de nim sobre pupas das moscas mencionadas em condições de laboratório e de campo. A redução da emergência das moscas das pupas tratadas com 0,6 por cento de óleo de nim foi de 95,6 por cento em condições de laboratório e 94,5 por cento quando aplicado ao solo onde descansam os animais durante a noite. A aplicação do óleo de nim a 0,6 por cento no solo é uma alternativa viável para controle das pupas de L. cuprina, C. megacephala, C. hominivorax e M. domestica.

Flies Lucilia cuprina, Chrysomya megacephala, Cochliomyia hominivorax and Musca domestica, are among the most important arthropod pests of livestock, which produce myiasis and carry pathogenic microorganisms. The purpose of this study was to test of efficacy of various concentrations of neem oil on pupae of the above mentioned flies under laboratory and in field conditions. Reduction of fly emergence from treated pupae with 0.6 per cent of neem oil was 95.6 per cent under laboratory conditions and 94.5 per cent when applied on the soil, where the cattle rest during the night. The application of neem oil at 0.6 per cent on the soil is a viable alternative for controlling pupae of L. cuprina, C. megacephala, C. hominivorax and M. domestica.