Exchange of viral segments between one or more influenza virus subtypes can contribute to a shift in virulence and adaptation to new hosts. Among several influenza subtypes, H9N2 is widely circulating in poultry populations worldwide and has the ability to infect humans. Here, we studied the reassortant compatibility between chicken H9N2 with N1-N9 gene segments of wild bird origin, either with an intact or truncated stalk. Naturally occurring amino acid deletions in the NA stalk of the influenza virus can lead to increased virulence in both mallard ducks and chickens. Our findings show extended genetic compatibility between chicken H9Nx gene segments and the wild-bird NA with and without 20 amino acid stalk deletion. Replication kinetics in avian, mammalian and human cell lines revealed that parental chH9N2 and rH9N6 viruses with intact NA-stalk replicated significantly better in avian DF1 cells compared to human A549 cells. After introducing a stalk deletion, an enhanced preference for replication in mammalian and human cell lines could be observed for rH9N2(Delta)(H6), rH9N6(Delta) and rH9N9(Delta) compared to the parental chH9N2 virus. This highlights the potential emergence of novel viruses with variable phenotypic traits, warranting the continuous monitoring of H9N2 and co-circulating subtypes in avian hosts.

Shrews are primary hosts for mammalian hantaviruses and are thus considered to be important reservoirs for viruses, similar to rodents and bats. To explore the diversity of hantaviruses in Swedish common shrews (Sorex araneus), we investigated lung tissues from shrews collected between 2015 and 2017. The collection took place at three separate locations in south-central Sweden. Screening for hantaviruses was performed using two different approaches: i) A total of 110 common shrews were investigated for hantaviruses by a pan-hanta L-gene RT-PCR, and Sanger sequencing was performed on the 13 positive samples. ii) In addition, 88 RNA samples were pooled into eight libraries subjected to RNA sequencing. The RNA sequencing data analysis, which focused specifically on identifying hantaviruses, revealed two divergent hantaviruses: the complete genome of an Altai-like virus and the partial genome of Seewis virus. Evolutionary analysis revealed that Swedish Altai-like viruses are closely related to Russian Altai viruses but distinct from Finnish strains. On the contrary, Swedish Seewis virus shares closer ancestry with Finnish Seewis virus strains. Given that these viruses were identified in several libraries, Seewis virus and Altai virus are likely circulating in Swedish common shrews. Supported by earlier studies, common shrews are probably the natural host for at least these two distinct hantaviruses.

In 2018, a local case of nephropathia epidemica was reported in Scania, southern Sweden, more than 500 km south of the previously known presence of human hantavirus infections in Sweden. Another case emerged in the same area in 2020. To investigate the zoonotic origin of those cases, we trapped rodents in Ballingsl & ouml;v, Norra Sandby, and S & ouml;rby in southern Sweden during 2020-2021. We found Puumala virus (PUUV) in lung tissues from 9 of 74 Myodes glareolus bank voles by screening tissues using a hantavirus pan-large segment reverse transcription FOR. Genetic analysis revealed that the PUUV strains were distinct from those found in northern Sweden and Denmark and belonged to the Finnish PUUV lineage. Our findings suggest an introduction of PUUV from Finland or Karelia, causing the human PUUV infections in Scania. This discovery emphasizes the need to understand the evolution, cross-species transmission, and disease outcomes of this newly found PUUV variant.