Wanaratana S, Panyim S, Pakpinyo S
Department of Veterinary Medicine, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
Med Vet Entomol. 2011 Mar;25(1):58-63. doi: 10.1111/j.1365-2915.2010.00928.x. Epub 2010 Dec 1.
The objective of the present study was to determine the potential for house flies (Musca domestica L.) (Diptera: Muscidae) to harbour the avian influenza (AI) H5N1 virus. Laboratory-reared flies were experimentally fed with a mixture containing the AI virus. Exposed flies were washed with brain-heart infusion broth and followed by 70% alcohol before preparation of whole fly homogenate. The homogenate was inoculated into six 10-day-old embryonated chicken eggs (ECEs). Allantoic fluids were collected to determine the virus using the haemagglutination (HA) test, reverse transcription-polymerase chain reaction (RT-PCR) or quantitative real-time RT-PCR (RRT-PCR). In the first experiment, ECEs that were inoculated with the 50 AI virus exposed fly homogenates died within 48 h and HA and RT-PCR were positive for AI virus. In the second experiment, ECEs that were inoculated with only one fly died with positive HA test and RT-PCR. In the last experiment, a group of exposed flies was collected at 0, 6, 12, 24, 36, 48, 72 and 96 h post-exposure. Fly homogenates of each time point were tested by virus titration in ECEs and RRT-PCR. Virus titres declined in relation to exposure time. Furthermore, RRT-PCR results were positive at any time point. The present study shows that the flies may harbour the AI virus and could act as a mechanical vector of the AI virus.
本研究的目的是确定家蝇(Musca domestica L.)(双翅目:蝇科)携带禽流感(AI)H5N1病毒的可能性。在实验室饲养的家蝇被用含有禽流感病毒的混合物进行实验性喂食。在制备家蝇全虫匀浆之前,将接触过病毒的家蝇先用脑心浸液肉汤冲洗,然后再用70%酒精冲洗。将匀浆接种到6个10日龄的鸡胚(ECEs)中。收集尿囊液,使用血凝(HA)试验、逆转录-聚合酶链反应(RT-PCR)或定量实时RT-PCR(RRT-PCR)来检测病毒。在第一个实验中,接种了50份接触过禽流感病毒的家蝇匀浆的鸡胚在48小时内死亡,HA和RT-PCR检测禽流感病毒呈阳性。在第二个实验中,仅接种一只家蝇的鸡胚死亡,HA试验和RT-PCR检测呈阳性。在最后一个实验中,在接触病毒后的0、6、12、24、36、48、72和96小时收集一组接触过病毒的家蝇。每个时间点的家蝇匀浆通过在鸡胚中进行病毒滴定和RRT-PCR进行检测。病毒滴度随接触时间下降。此外,在任何时间点RRT-PCR结果均为阳性。本研究表明,家蝇可能携带禽流感病毒,并可能作为禽流感病毒的机械传播媒介。