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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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... Because C. grahami colonizes decomposing materials and is synanthrophropic, it can transmit human and animal pathogens from decomposing animals to individuals living in the surrounding area. One pathogen transmitted by this specie is the avian influenza virus (H5N1) (Tsuda et al. 2009, Sawabe et al. 2006. Other bacteria such as ...
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Se presentan resultados de la sucesión ecológica de insectos asociados a cadaveres de Cerdos (Sus scrofa) en Saltillo, Coahuila así como el desarrollo de Phormia regina bajo condiciones de laboratorio y su validación en campo.
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Chapter
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Field study in Hachijo Island, Tokyo, showed Calliphora nigribarbis Vollenhoven was short-day univoltine insect whose development continued for 6 months (December to May). Adults appeared from November to June in towns (alt. 50m) and from October to July in the montane moss forests of Mt. Hachijo-Fuji (alt. 850m). The number of flies at the upland gradually increased during February to April and then decreased. After the rainy season in June, no adults were trapped throughout summer in both lowlands and uplands. No developing stages were also observed from June to November in both habitats. The first appearance of aged males and females in moss forests in October, suggests that adults aestivate there. After aestivation some females migrate from uplands to lowlands for spawning eggs. The lowland population emerged in spring migrates to uplands for aestivation. This up- and down seasonal migration was confirmed by the release and recapture of marked flies in the field experiment made during the same period. Adults seem to escape from lowlands in order to maintain diapause.
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An investigation was made to know the seasonal fluctuation of Aldrichina grahami (Aldrich) in Aogashima Island, Tokyo, from December 1982 to December 1984. A trap baited with horse flesh was set at a station on a hill at 300m above sea level, and the numbers of flies trapped were counted once a month. Adult flies were captured from November to June or July, but none was seen for succeeding 3-4 months. It is not clear that females captured in autumn could aestivate in the island or migrated from the nearest Hachijo Island.
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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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Eleven species of the calyptrate muscoid flies were captured on the ships which were located at the ocean weather stations or voyaged on the Pacific Ocean and East China Sea. Calliphora nigribarbis Vollenhoven and Aldrichina grahami (Aldrich) visited the weather ships at the stations which are located on the Pacific Ocean and East China Sea, 300 to 450km off Kyushu, Japan. Bengalia torosa (Wiedemann) was captured on a ship voyaged more than 100km from Taiwan where this fly is commonly found. These three species and Delia platura (Meigen) are considered to be transoceanic migratory insects, because they had more or less heavily damaged wings caused by a long-distant flight. The following 7 species, Lucilia cuprina, L. sericata, Musca domestica, Muscina stabulans, Atherigona varia, Lispe assimilis and Lispocephala miki, are considered to have moved with the ships, because they have had complete wings.
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In the autumn of 1995,we had an opportunity to witness and evaluate the importance of the mass, probably transoceanic, migratory flight and landing of Calliphora nigribarbis Vollenhoven in the campus of Institute of Tropical Medicine, Nagasaki University, Nagasaki, Kyushu, western Japan. On October 25 (14 : 30 to 17 : 30,fine) and 26 (10 : 30 to 17 : 40,fine), they were seen flying in large numbers from the northwest to southeast across the top of the laboratory building (about 10m in height). They flew continuously in the direction of the wind and we caught some of them at the top of the building. They flew vigorously, with many flying well beyond the reach of our insect nets and without changing course. Some flies reaching the building in the evening landed on the concrete wall of the building. The migratory flights were simultaneously observed in Fukuoka where the flies were seen flying from the north across Tsushima-Kaikyo Strait. Fukuoka and Nagasaki are situated about 250 to 300km southeast of the Korean Penninsula. It could be inferred that these flies migrate across from the Asian continent to the main island Kyushu. C. nigribarbis has been considered to migrate altitudinally in the mainland Japan. However, the number of flies found in autumn (November) in Kyushu appears to be also increased by the transoceanic migration.
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.