Natalie N. Tarbuck

Since 2024, highly pathogenic influenza A(H5N1) viruses have spread extensively among U.S. dairy cattle, where they replicate efficiently in the mammary gland and are shed at high titers in milk. To directly assess susceptibility of commercial swine populations to bovine-derived H5N1 virus, lactating sows with prior influenza virus vaccination histories representative of U.S. commercial swine production systems were inoculated via the intramammary route and co-housed with their 1-week-old piglets to evaluate disease outcomes, viral replication, and potential for vertical transmission. Intramammary inoculation of lactating sows resulted in sustained viral RNA shedding in milk, while piglets exhibited sporadic oral viral RNA positivity that mirrored viral kinetics in milk. Lesions in mammary tissue and viral antigen staining, as well as development of neutralizing antibody responses and changes in milk color and consistency, further confirmed infection in the sows. Despite these molecular findings, none of the animals developed overt clinical disease, and respiratory involvement was not noted during the study period. Collectively, we demonstrate that intramammary exposure results in productive influenza A(H5N1) virus infection in lactating sows despite their vaccination histories, indicating the potential threat of viral spillover into commercial swine populations. The clinically inapparent nature of infection presents a risk of subclinical spread and underscores the importance of expanding viral surveillance to swine.

American mink (Neogale vison) are susceptible to SARS-CoV-2, but little is known about virus circulation in mink since 2021. Here, in the first active surveillance study of SARS-CoV-2 in apparently healthy farmed mink in the United States, we find a ~0.9% (760/85,656) RT-PCR positivity rate among nasal swabs collected in 18 farms across six states during 2022–2023. Phylogenetic analysis of 293 viral genome sequences shows that human-adapted SARS-CoV-2 variants (e.g., Omicron) repeatedly spill over into mink. Surprisingly, the detection of a Delta lineage virus (AY.39) on a mink farm four months after its last detection in humans within the same state suggests prolonged unsampled transmission. The spread of mink-adapted AY.39 viruses from a mink farm to neighboring free-ranging white-tailed deer represents a rare instance of SARS-CoV-2 transmission between livestock and wildlife. These findings demonstrate the value of active surveillance for identifying subclinical infections and interspecies transmission between humans, mink, and wildlife.

Highly pathogenic avian influenza A(H5N1) virus exhibits a strong tropism for the bovine mammary gland, challenging our understanding of influenza A virus host range and tissue specificity. We performed experimental studies with an influenza A(H5N1) B3.13 genotype virus in female lactating dairy cattle to define the infectious dose, routes of exposure, and factors linked to morbidity and mortality. Here, we demonstrate that intramammary inoculation with as few as 10 TCID₅₀ establishes a robust infection and shedding of high-titer virus in milk. Despite this low infectious dose, H5N1 does not readily transmit via contaminated milking equipment and close contact with infected animals. High-dose intramammary exposure results in severe disease and mortality, while respiratory and oral exposures are less likely to establish productive infection and associated morbidity. This study challenges current hypotheses of H5N1 transmission on dairy farms, raising important questions about potential agent, host, or environmental cofactors contributing to viral spread.

Ongoing transmission of influenza A virus (H5N1) in U.S. dairy cattle threatens both animal and human health, underscoring the need to understand the durability of host immunity against reinfection with evolving genotypes. We challenged naive and convalescent cows, infected one year prior with H5N1 genotype B3.13, with either homologous B3.13 or heterologous D1.1 genotype virus. Homologous rechallenge resulted in complete clinical protection with no infectious viral shedding. Conversely, heterologous rechallenge led to transient clinical disease and limited infectious viral shedding. Convalescent cows experienced significantly milder disease than naive cows, which developed severe illness with high viral shedding and required early euthanasia, regardless of the strain. These findings indicate that naturally acquired immunity offers strong protection against severe illness but may allow silent transmission of divergent strains. Therefore, natural herd immunity alone is unlikely to eliminate the virus; controlling H5N1 in cattle will likely require vaccination strategies that address viral evolution.

The discovery that highly pathogenic avian influenza A(H5N1) virus exhibits a strong tropism for the bovine mammary gland1–4 represents a major shift in our understanding of influenza A virus host range and tissue specificity. We conducted a comprehensive series of experimental studies with influenza A(H5N1) B3.13 genotype in lactating dairy cattle to address several key questions related to the viral dose required to establish infection, routes of exposure that lead to transmission, and factors contributing to the morbidity and mortality observed on farms. We demonstrate that intramammary exposure to as few as 10 TCID50 is sufficient to establish robust infection, shedding of high viral titers in milk, and clinical mastitis. Despite evidence of such a low infectious dose, we were unable to recapitulate transmission to sentinel cows via contaminated milking equipment and close contact with infected animals under experimental conditions. High-dose intramammary exposure to influenza A(H5N1) drives severe clinical outcomes and mortality observed in dairy cows on-farm, while respiratory and oral exposure are less likely to establish productive infection and associated morbidity. This study challenges current hypotheses of influenza A(H5N1) transmission on dairy farms5,6, raising important questions about potential agent, host, or environmental cofactors that contribute to the spread of the virus.