U.S. Geological Survey, Alaska Science Center, Anchorage, Alaska, USA.
Appl Environ Microbiol. 2013 Dec;79(23):7249-55. doi: 10.1128/AEM.02439-13. Epub 2013 Sep 13.
The principal mode of avian influenza A virus (AIV) transmission among wild birds is thought to occur via an indirect fecal-oral route, whereby individuals are exposed to virus from the environment through contact with virus-contaminated water. AIV can remain viable for an extended time in water; however, little is known regarding the influence of the biotic community (i.e., aquatic invertebrates) on virus persistence and infectivity in aquatic environments. We conducted laboratory experiments to investigate the ability of an aquatic filter-feeding invertebrate, Daphnia magna, to accumulate virus from AIV-dosed water under the hypothesis that they represent a potential vector of AIV to waterfowl hosts. We placed live daphnids in test tubes dosed with low-pathogenicity AIV (H3N8 subtype isolated from a wild duck) and sampled Daphnia tissue and the surrounding water using reverse transcription-quantitative PCR (RT-qPCR) at 3- to 120-min intervals for up to 960 min following dosing. Concentrations of viral RNA averaged 3 times higher in Daphnia tissue than the surrounding water shortly after viral exposure, but concentrations decreased exponentially through time for both. Extracts from Daphnia tissue were negative for AIV by cell culture, whereas AIV remained viable in water without Daphnia present. Our results suggest daphnids can accumulate AIV RNA and effectively remove virus particles from water. Although concentrations of viral RNA were consistently higher in Daphnia tissue than the water, additional research is needed on the time scale of AIV inactivation after Daphnia ingestion to fully elucidate Daphnia's role as a potential vector of AIV infection to aquatic birds.
野生鸟类中禽流感病毒(AIV)的主要传播模式被认为是通过间接的粪-口途径发生的,在此过程中,个体通过接触受病毒污染的水而暴露于环境中的病毒。AIV 在水中可以长时间保持存活;然而,关于生物群落(即水生无脊椎动物)对水生环境中病毒持久性和感染力的影响,人们知之甚少。我们进行了实验室实验,以调查一种水生滤食性无脊椎动物——大型溞(Daphnia magna)从 AIV 处理水中积累病毒的能力,其假设是它们代表了 AIV 向水禽宿主传播的潜在载体。我们将活的大型溞放入装有低致病性 AIV(从野鸭中分离出的 H3N8 亚型)的试管中,并在给药后 3 至 120 分钟的时间间隔内使用反转录定量 PCR(RT-qPCR)每隔 3 至 120 分钟对大型溞组织和周围水进行采样,时间长达 960 分钟。病毒暴露后不久,大型溞组织中的病毒 RNA 浓度平均比周围水高出 3 倍,但随着时间的推移,两种浓度都呈指数下降。从大型溞组织中提取的病毒 RNA 经细胞培养呈 AIV 阴性,而在没有大型溞存在的情况下,AIV 在水中仍具有活性。我们的结果表明,大型溞可以积累 AIV RNA,并有效地从水中去除病毒颗粒。尽管大型溞组织中的病毒 RNA 浓度始终高于水,但需要进一步研究 AIV 在大型溞摄入后失活的时间尺度,以充分阐明大型溞作为 AIV 感染水禽的潜在载体的作用。