Estimation of Avian Influenza Viruses in Water Environments of Live Poultry Markets in Changsha, China, 2014 to 2018.

In routine surveillance for avian influenza viruses (AIVs) in the environments of live poultry markets (LPMs), certain samples were positive for AIVs type A while negative for subtypes (e.g., H5, H7, and H9). However, little attention has been paid to these unsubtyped AIVs samples. To reveal the dynamic distribution and molecular characteristics of AIVs, especially the unsubtyped AIVs, we reported and analyzed 1969 samples collected from the water environments of LPMs in Changsha, China, from January 2014 to November 2018. Our results revealed that 1504 (76.38%) samples were positive for AIV type A. Of these samples, the predominant hemagglutinin (HA) subtype was H9, followed by H5 and H7 (P < 0.05). The positive rate of H5 subtype in water environmental samples exhibited seasonality, which reached a peak in each winter-spring season from January 2014 to March 2017. The positive rates of AIVs (including type A, subtype H9, and mixed subtype H5/H7/H9) in non-central-city regions were higher than that in the central-city regions (P < 0.05). Notably, 161 unsubtyped AIVs samples were detected during the routine surveillance. However, subtyping with the commercial kit further identified eight different HA and seven different neuraminidase subtypes. Analyses unraveled that further subtyped AIVs H1, H6, and H11 had only one basic amino acid (R or K) at the cleavage site and residues Q226 and G228 at the receptor-binding associated sites. Overall, in addition to H5, H7, and H9 subtypes, we should also pay attention to unsubtyped AIVs samples during the routine surveillance for AIVs in the environments of LPMs.