Development and validation of concentration methods for sensitive detection of avian influenza virus in environmental water

Wild birds, especially aquatic birds, are the natural reservoir of avian influenza virus (AIV), and many kinds of water body can be contaminated with feces of these birds. Seasonally, AIVs can be dissolved in the environmental water from the feces of the infected birds, and this water can be a target for viral detection and identification. In this study, we employed and tested three different filters for concentrating AIV, and it was shown that high concentration factor in terms of viral density could be achieved with viral samples diluted with natural water. Wild bird fecal samples containing low pathogenicity H5 AIVs were successfully concentrated with the adsorption and elution method using mixed cellulose esters membrane; the recovery rate of virus was 35.5 % and the concentration factor was about 50 on average. For the larger volume of water sample, we proved that an inline disposable filter with high surface area, 300 cm2, has a comparable concentration factor to the adsorption and elution method and the filter could be used in the field conveniently by being plugged into peristaltic pump. These validated methods for water sampling may be used as a supplementary for virological surveillance on wild migratory birds or during the epidemiological investigation on the environment near affected premises by AIV.