We isolated two highly pathogenic avian influenza viruses (HPAIVs) of subtype H5N8 clade 2.3.4.4b from falcated duck (Anas falcata) feces and environmental water collected at an overwintering site in Japan. Our isolates were almost genetically identical to each other and showed high genetic similarity with H5N8 HPAIVs recently isolated in South Korea, a distant part of Japan, and European countries. These results suggest the potential role of falcated ducks in the dissemination of HPAIVs.

Highly pathogenic avian influenza viruses (HPAIVs) have caused outbreaks in both domestic and wild birds during the winter seasons in several countries in the Northern Hemisphere, most likely because virus-infected wild ducks overwinter and serve as the primary source of infection for other birds in these countries. Several chemical disinfectants are available to deactivate these viruses outside a living organism. However, their virucidal activity is known to be compromised by various factors, including temperature and contamination with organic matter. Hence, the effectiveness of virucidal activity under winter field conditions is crucial for managing HPAIV outbreaks. To investigate the impact of the winter field conditions on the virucidal activity of disinfectants against AIVs, we assessed the stability of the virucidal activity of seven representative disinfectants that are commercially available for poultry farms in Japan against both LPAIVs and HPAIVs under cold and/or organic contamination conditions. Of the seven disinfectants examined, the ortho-dichlorobenzene/cresol-based disinfectant exhibited the most consistent virucidal activity under winter field conditions, regardless of the virus pathogenicity or subtype tested.

Recent global outbreaks of highly pathogenic avian influenza viruses (HPAIVs) of the H5N8 subtype in poultry and wild birds have raised concerns about animal and human health, particularly after its first evidence of zoonotic transmission from birds to humans. Here, we report a lethal infection with the H5N8 HPAIV in a mandarin duck that had previously demonstrated resistance to the H5N8 HPAIV infection. In addition, we revealed that the isolated virus was a genetic reassortant between the existing H5N8 HPAIV and LPAIV(s). Although further studies are warranted to assess the impact of the genetic reassortment on virus pathogenicity, the potential role of mandarin ducks in HPAIV dissemination should be re-evaluated.

Porcine reproductive and respiratory syndrome (PRRS), which is caused by the porcine reproductive and respiratory syndrome virus (PRRSV), has a significant impact on the global pork industry. It results in reproductive failure in sows and respiratory issues in pigs of all ages. Despite the availability of vaccines, controlling the PRRSV remains challenging, partly owing to the limitations of cell culture systems. Current methods largely rely on primary porcine alveolar macrophages (PAMs), which must be harvested from piglets and have limited proliferative capacity. Although some simian cell lines support PRRSV replication, their inability to express porcine CD163, which is a key receptor for PRRSV entry, compromises their effectiveness, because the virus replicates differently in these non-target cells. To address these issues, we established an immortalized PAM cell line, PAM-T43, using SV40 large T antigen for immortalization and porcine serum as a culture supplement. PAM-T43 cells maintain essential macrophage functions, including CD163 expression and phagocytic activity, and exhibit high sensitivity to the PRRSV, efficiently supporting viral replication. This novel cell line offers significant potential for advancing PRRSV research, particularly in vaccine development and field strain isolation, by overcoming the limitations of current systems.

Avian influenza (AI) is an extremely contagious viral disease of domestic and wild birds that can spread rapidly among bird populations, inducing serious economic losses in the poultry industry. During the winter season 2021–2022, we isolated seventeen highly pathogenic avian influenza (HPAI) H5N8 viruses from outbreaks involving ducks in Egypt, occurring in both backyard and farm settings. The aim of this study was to pinpoint genetic key substitutions (KSs) that could heighten the risk of a human pandemic by influencing the virus’s virulence, replication ability, host specificity, susceptibility to drugs, or transmissibility. To understand their evolution, origin, and potential risks for a human pandemic, whole-genome sequencing and phylogenetic analysis were conducted. Our analysis identified numerous distinctive mutations in the Egyptian H5N8 viruses, suggesting potential enhancements in virulence, resistance to antiviral drugs, and facilitation of transmission in mammals. In this study, at least five genotypes within one genome constellation of H5N8 viruses were identified, raising concerns about the potential emergence of novel viruses with altered characteristics through reassortment between different genotypes and distinct groups. These findings underscore the role of ducks in the virus’s evolutionary process and emphasize the urgent need for enhanced biosecurity measures in domestic duck farms to mitigate pandemic risk.

The Izumi plain in the Kagoshima Prefecture, Japan, is known as an overwintering site for more than 30,000 migratory waterfowl, including endangered crane species. We previously reported that environmental water samples, from artificial wet paddies created as crane roost sites on the Izumi plain, are useful for avian influenza virus (AIV) surveillance. During the 2019/20 winter season, we collected 238 water samples from the crane roost sites and isolated 22 AIVs of six subtypes: one H1N1, one H3N2, seven H3N8, four H4N6, nine H6N6, and one H11N2 subtypes. Genetic analyses revealed that AIVs of the same subtype isolated from the Izumi plain during a single winter season exhibited multiple genetic constellations. Furthermore, phylogenetic analyses suggested that our H3N2 isolate may be a genetic reassortant between close relatives to our H3N8 and H11N2 isolates. Our study highlighted the importance of monitoring AIV circulation to better understand AIV ecology in migratory waterfowl populations.