A One Health Investigation into H5N1 Avian Influenza Virus Epizootics on Two Dairy Farms

Ismaila Shittu, Diego Silva, Judith U. Oguzie, Lyudmyla V. Marushchak, Gene G. Olinger, John A. Lednicky, Claudia M. Trujillo-Vargas, Nicholas E. Schneider, Haiping Hao, Gregory C. Gray
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Abstract

Background: In early April 2024 we studied two Texas dairy farms which had suffered incursions of H5N1 highly pathogenic avian influenza virus (HPAIV) the previous month. Methods: We employed molecular assays, cell and egg culture, Sanger and next generation sequencing to isolate and characterize viruses from multiple farm specimens (cow nasal swab, milk specimens, fecal slurry, and a dead bird). Results: We detected H5N1 HPAIV in 64% (9/14) of milk specimens, 2.6% (1/39) of cattle nasal swab specimens, and none of 17 cattle worker nasopharyngeal swab specimens. We cultured and characterized virus from eight H5N1-positive specimens. Sanger and next-generation sequencing revealed the viruses were closely related into other recent Texas epizootic H5N1 strains of clade 2.3.4.4b. Our isolates had multiple mutations associated with increased spillover potential. Surprisingly, we detected SARS-CoV-2 in a nasal swab from a sick cow. Additionally, 14.3% (2/14) of the farm workers who donated sera were recently symptomatic and had elevated neutralizing antibodies against a related H5N1 strain. Conclusions: While our sampling was limited, these data offer additional insight into the large H5N1 HPAIV epizootic which thus far has impacted at least 96 cattle farms in twelve US states. Due to fears that research might damage dairy businesses, studies like this one have been few. We need to find ways to work with dairy farms in collecting more comprehensive epidemiological data that are necessary for the design of future interventions against H5N1 HPAIV on cattle farms.
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对两个奶牛场 H5N1 禽流感病毒疫情的统一健康调查
背景:2024 年 4 月初,我们对德克萨斯州的两个奶牛场进行了研究,这两个奶牛场在前一个月曾遭遇 H5N1 高致病性禽流感病毒(HPAIV)入侵。研究方法我们采用分子检测、细胞和卵培养、桑格测序和新一代测序等方法,从多个牧场标本(牛鼻拭子、牛奶标本、粪便和一只死鸟)中分离病毒并确定其特征。结果:我们在 64%(9/14)的牛奶标本、2.6%(1/39)的牛鼻拭子标本和 17 份牛工鼻咽拭子标本中均未检测到 H5N1 高致病性禽流感病毒。我们对 8 份 H5N1 阳性标本中的病毒进行了培养和鉴定。桑格测序和下一代测序显示,这些病毒与德克萨斯州最近发生的其他 H5N1 流行病毒株 2.3.4.4b 支系密切相关。我们的分离株有多种突变,具有更高的溢出潜力。令人惊讶的是,我们在一头病牛的鼻拭子中检测到了 SARS-CoV-2 病毒。此外,14.3%(2/14)的农场工人最近出现了症状,并对相关的 H5N1 株产生了较高的中和抗体。结论:虽然我们的取样有限,但这些数据提供了对大规模 H5N1 高致病性禽流感疫情的更多了解,迄今为止,该疫情已影响到美国 12 个州的至少 96 个养牛场。由于担心研究可能会损害乳制品企业,像这样的研究一直很少。我们需要设法与奶牛场合作,收集更全面的流行病学数据,这些数据对于设计未来针对养牛场 H5N1 高致病性禽流感的干预措施十分必要。
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