Hubbard Laura E, Stelzer Erin A, Poulson Rebecca L, Kolpin Dana W, Szablewski Christine M, Givens Carrie E
U.S. Geological Survey, Upper Midwest Water Science Center, 1 Gifford Pinchot Drive, Madison, WI 53726, USA.
U.S. Geological Survey, Ohio-Kentucky-Indiana Water Science Center, 6460 Busch Blvd, Ste 100, Columbus, OH 43229, USA.
Viruses. 2024 Dec 10;16(12):1898. doi: 10.3390/v16121898.
Since late 2021, outbreaks of highly pathogenic avian influenza virus have caused a record number of mortalities in wild birds, domestic poultry, and mammals in North America. Wetlands are plausible environmental reservoirs of avian influenza virus; however, the transmission and persistence of the virus in the aquatic environment are poorly understood. To explore environmental contamination with the avian influenza virus, a large-volume concentration method for detecting infectious avian influenza virus in waterbodies was developed. A variety of filtering, elution, and concentration methods were explored, in addition to testing filtering speeds using artificially amended 20 L water matrices (deionized water with sterile dust, autoclaved wetland water, and wetland water). The optimal protocol was dead-end ultrafiltration coupled with salt solution elution and centrifugation concentration. Using this method, infectious virus was recovered at 1 × 10 50% egg infectious dose per milliliter (EID/mL), whereas viral RNA was detected inconsistently down to 1 × 10 EID/mL. This method will aid in furthering our understanding of the avian influenza virus in the environment and may be applicable to the environmental detection of other enveloped viruses.
自2021年末以来，高致病性禽流感病毒的爆发已导致北美野生鸟类、家禽和哺乳动物的死亡数量创下纪录。湿地可能是禽流感病毒的环境储存库；然而，人们对该病毒在水生环境中的传播和持续存在了解甚少。为了探究禽流感病毒对环境的污染情况，开发了一种用于检测水体中传染性禽流感病毒的大容量浓缩方法。除了使用人工改良的20升水基质（含有无菌灰尘的去离子水、高压灭菌的湿地水和湿地水）测试过滤速度外，还探索了各种过滤、洗脱和浓缩方法。最佳方案是死端超滤结合盐溶液洗脱和离心浓缩。使用该方法，每毫升可回收1×10 50%鸡胚感染剂量（EID/mL）的传染性病毒，而病毒RNA的检测下限为1×10 EID/mL，结果并不一致。该方法将有助于加深我们对环境中禽流感病毒的理解，并且可能适用于其他包膜病毒的环境检测。
