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Detection and isolation of highly pathogenic H5N1 avian influenza A viruses from blow flies collected in the vicinity of an infected poultry farm in Kyoto, Japan, 2004

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During the outbreak of highly pathogenic avian influenza that occurred in Tamba Town, Kyoto Prefecture in 2004, a total of 926 flies were collected from six sites within a radius of 2.3 km from the poultry farm. The H5 influenza A virus genes were detected from the intestinal organs, crop, and gut of the two blow fly species, Calliphora nigribarbis and Aldrichina grahami, by reverse transcription-polymerase chain reaction for the matrix protein (M) and hemagglutinin (HA) genes. The HA gene encoding multiple basic amino acids at the HA cleavage site indicated that this virus is a highly pathogenic strain. Based on the full-length sequences of the M, HA, and neuraminidase (NA) segments of virus isolates through embryonated chicken eggs, the virus from C. nigribarbis (A/blow fly/Kyoto/93/2004) was characterized as H5N1 subtype influenza A virus and shown to have > 99.9% identities in all three RNA segments to a strain from chickens (A/chicken/Kyoto/3/2004) and crows (A/crows/Kyoto/53/2004) derived during this outbreak period in Kyoto in 2004. Our results suggest it is possible that blow flies could become a mechanical transmitter of H5N1 influenza virus.
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... Insects such as mosquitoes, blow flies and house flies are suggested as possible vectors of HPAIv. During a HPAI H5N1 epidemic on poultry farms in Japan, flies were collected from six sites within a radius of approximately 2 km from infected poultry farms [31]. HPAI H5N1 virus-identical to the virus strain derived from chickens, large-billed crows (Corvus macrorhynchos) and Black crows in the epidemic area-was detected in the intestinal organs, crop and gut of two blow fly species by RT-PCR, suggesting that blow flies can be a mechanical transmitter of HPAIv. ...
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The breeding and emerging periods of Aldrichina grahami (Aldrich) were studied along with the fluctuation of adults for tracing its life cycle by using both baited and emergence traps in the backyard of the National Institute of Health, Tokyo, from December 1981 to March 1983. Seasonal activity of the adult in each year was similar, which confirmed the previous works reported from the same locality. A number of both sexes were trapped from March to July, and a large amount of females were caught in September to December. The first flight in early fall represents only aged females attracted. The breeding stage first appeared in the beginning of September and ended in the middle of coming May. Early fall generation derived from eggs deposited in September quickly grew up and emerged as adults in late fall (October to November). Prepupae or pupae of late fall generation derived from eggs in November to December became dormant during winter. After the dormancy, most adults emerged in March, though a few appeared in the end of January and the beginning of February. Spring generation from eggs in February to May normally developed within relatively short periods (0.5-1.5 months). No developing stages were observed from late May to early September, although male and female adults were trapped until the end of July. From the above, it is concluded that this blow fly can develop in the Tokyo area during 8.5 months from September to the middle of May excepting the coldest season in which dormant prepupae and pupae are dominant. The first appearance of aged females in September may suggest the adult aestivation in Aldrichina grahami.
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An investigation was made to know the life history of Aldrichina grahami (Aldrich, 1930) in Hachijo Island, Tokyo, from February 1981 to May 1982. A trap baited with horse meat was set at five different altitudes, 50m, 200m, 400m, 600m and 800m, of Mt. Hachijo-Fuji. The number of flies captured was counted once a month. The larvae and pupae which had bred from meat in the trap were also counted and transferred into a separate trap for emergence. The number of the flies gradually increased from foot to top of the mountain during March to May. Following the rainy season in June, the number decreased and only one female was trapped at alt. 600m during August to September. Some females were captured again at alt. 600m and 800m in October. This suggests that some of mated females can estivate in the mountainous area above alt. 600m where it is usually cool and foggy during summer and mossforests develop. A few adults and a number of pupae were found even in winter in the trap of station 600m. The upper region of the mountain may be considered as a primary habitat of this blow fly. After estivation some females probably migrated for spawning eggs from the mountainous habitat and appeared in the town at alt. 50m. They may be successful in finding enough foods and breeding places around the human dwellings in winter and spring. The 74% of the flies were trapped in a temperature range of 2.0-9.0℃ in minimum and 16.5-23.0℃ in maximum and in a range of day length 10 : 30-14 : 20. This also suggests that the adult has a preference for such climate as being rather cool before sunrise and warm in daytime in spring and autumn.
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Mark and recapture experiments of blow flies, Calliphora nigribarbis Vollenhoven and Aldrichina grahami (Aldrich), were attempted on Hachijo Island, Tokyo, in the autumn of 1983 and the spring of 1984 to investigate their seasonal migration. One female C. nigribarbis released at Station (St.) M (altitude 700m) in the moss forest of Mt. Hachijo-Fuji (854m) at the end of October 1983 was recaptured at St. 1 (50m) in the town at the end of November. At the end of March 1984,a total of 841 flies of both sexes of C. nigribarbis and A. grahami were marked and released at St. T (50m). One male and one female C. nigribarbis and one male A. grahami were recaptured at St. G (600m), St. 3 (400m) and St. 2 (200m), respectively, at the end of April. One female C. nigribarbis released on 24-25 May 1984 at St. T was trapped at St. 4 (600m) on 29 May 1984. The result of mark and recapture experiments confirmed the possibility that adults emerged in the town migrate from the lowland to the moss forest of Mt. Hachijo-Fuji before summer. Some of them most probably return to the lowland in autumn.
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Chapter
The calyptrate Diptera are relatively homogenous in their appearance compared to the Nematocera, being mostly robust flies with conspicuous bristles — in fact the epitome of the popular idea of flies′. The 18 000 or so described species form a natural group (′subsection′) within the higher Diptera (i.e. the infraorder Muscomorpha). Their classification is described in brief in Chapter 3.