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ABSTRACT: To verify houseflies Musca spp. as the intermediate host of Thelazia callipaeda and reveal epidemiological situation of thelaziasis in Hubei province. Dogs eyes infected with T. callipaeda, 400 houseflies Musca and 259 fruitflies Amiota okadai in the city of Laohekou city (previously named as Guanghua county) of Hubei province had been investigated since September 2000. The newborn larvae of T. callipaeda from Laohekou suburbs were fed to houseflies Musca and A. okadai. Larvae used for the study were isolated from female T. callipaeda in laboratory and the susceptibility to houseflies Musca and A. okadai was observed. Twenty-one dogs from Laohekou, the original epidemic areas of thelaziasis were examined and 7 positive dogs in 21 (33.3%) and 11 T. callipaeda (9 females and 2 males) were identified. From 1975 to 2000, no thelaziasis cases were found through retrospective surveys. These 200 houseflies Musca and 135 A. okadai were dissected for examination but showed all negative with the infection. However, newborn larvae of T. callipaeda were used to experimentally infect 112 houseflies Musca and 84 A. okadai and all infected flies were examined on the 20th day after inoculation. As a consequence, houseflies Musca failed to be infected but 9 in 84 (10.7%) A. okadai were positive. 26 infective larvae of T. callipaeda were obtained and 21 of them were inoculated into right eye of one rabbit. The female worm began to produce newborn larvae in 37 days after infection and 3 adult T. callipaeda (two females and one male) were obtained. Fruitflies A. okadai from Hubei province were susceptible to T. callipaeda, which was similar to the result of experimental studies in Anhui province. This survey further confirmed that A. okadai was the intermediate host of T. callipaeda but not houseflies Musca. Infective resources (adult dogs, for instance) had been under controlled thus human thelaziasis had been eradicated in this rural area.