ETIDIOH - NEW

  Abstract The incursion of high pathogenicity avian influenza A virus (IAV) into US dairy cows is unprecedented in the era of molecular diagnosis and pathogen sequencing. This raises questions over the likelihood of further outbreaks and whether dairy cattle could be a mixing vessel for novel strains of IAV. Using a panel of BSL2-safe reassortant viruses representing clade 2.3.4.4b H5 epizootic lineages circulating since 2020, we found that a cow B3.13 isolate displayed enhanced replication in cow mammary gland cells , along with increased viral polymerase activity and stronger interferon antagonism in cow cells compared to an earlier EA-2020-C genotype virus. However, multiple avian and mammalian IAV strains , including other clade 2.3.4.4b high pathogenicity genotypes, were replication competent in bovine cells, particularly those of the mammary gland , suggesting that there is a diverse circulating IAV pool with the potential to infect cows. Moreover, we show that cow mammary c...

  Abstract The circulation of highly pathogenic H5 influenza A viruses in cattle , other mammals, and wildlife threatens animal and human health. To address this, we vaccinated heifer-calves with a deactivated rabies-virus-based H5 vaccine , which was well-tolerated and elicited neutralizing antibodies against both clade-1 and clade-2.3.4.4b H5N1 viruses, comparable to naturally H5-infected and convalescing cows. The immune responses to the vaccine platform were durable for at least 200 days and unaffected by preexisting RABV immunity. Source: npj Vaccines,  https://www.nature.com/articles/s41541-025-01238-2 ____

Abstract In early 2024, an unprecedented outbreak of H5N1 high pathogenicity avian influenza was detected in dairy cattle in the USA . As of mid-2025 the epidemic is ongoing, resulting in spillbacks into poultry, wild birds and other mammals including humans . Here, we present molecular and virological evidence that the cattle B3.13 genotype H5N1 viruses rapidly accumulated adaptations in polymerase genes that enabled better replication in bovine cells and tissues, as well as cells of other mammalian species including humans and pigs . We find evidence of several mammalian adaptations gained early in the evolution of these viruses in cattle including PB2 M631L , which is found in all cattle sequences, and PA K497R , which is found in the majority. Structurally, PB2 M631L maps to the polymerase-ANP32 interface, an essential host factor for viral genome replication . We show that this mutation adapts the polymerase to better interact with bovine ANP32 proteins , particularly ANP32A, and ...

  ABSTRACT Influenza A virus (IAV) infection of the respiratory tract can cause both respiratory and non-respiratory symptoms . Gastrointestinal (GI) symptoms such as diarrhea, vomiting, and abdominal pain can occur in persons with seasonal influenza A or novel IAV infections , but the extent to which IAVs can infect and replicate in GI tissues is understudied. The ongoing outbreak of A( H5N1 ) IAV in US dairy cattle associated with sporadic human infections has highlighted the potential public health threat posed by the introduction of infectious virus into materials that may be consumed by humans , such as milk. Here, we review epidemiologic reports documenting the frequency of GI complications in humans infected with seasonal and novel IAVs and present laboratory studies supporting the capacity of IAV to replicate in mammalian GI tissues , with an emphasis on A(H5N1) viruses. Studies assessing the ability of IAV to cause mammalian infection following consumption of virus-contain...

  ABSTRACT The mammary gland is an essential organ for milk production, providing essential immune and nutritional support to offspring and supplying dairy products for human consumption. In both humans and animals , the lactating mammary gland is susceptible to bacterial and viral infections , which can lead to mastitis and, in some cases, vertical transmission to offspring , with potential adverse effects on infant health. However, until recently, the role of respiratory viruses in mammary gland infection has been relatively understudied, particularly their ability to infect mammary epithelial cells and transmit through lactation. The recent emergence of highly pathogenic avian influenza H5N1 clade 2.3.4.4b in dairy cattle has demonstrated the virus’s capacity to replicate in the mammary gland, cause mastitis, and produce high viral loads in milk . This raises significant concerns about the potential for zoonotic transmission to humans and other animals in contact with infected d...

ABSTRACT The 2.3.4.4b clade highly pathogenic avian influenza H5N1 infected diverse non-human mammalian species , gained mammal-to-mammal transmission potential , and caused sporadic human infections . However, whether non-human mammals enable the human adaptation of 2.3.4.4b H5N1 to establish human infections is unclear. Gain-of-function research restrictions may hinder the assessment of 2.3.4.4b H5N1 human adaptations. Here, we tracked the evolution of 2.3.4.4b H5N1 that infected non-human mammals and evaluated their ability to gain human adaptations. The non-human mammal 2.3.4.4b H5N1 partly acquired classical human-adapting mutations , which are identical to the residues of H1N1pdm09 and seasonal human H3N2 viruses, while showing a few species-specific adaptations that might be potential barriers for successful human infections. The polymerase complex proteins , PA and PB2, acquired human adaptations in non-human mammals, with fox-infected viruses showing more positive selection in...

  ABSTRACT The H5N1 strain of influenza A virus (IAV) continues to cause severe infections in a range of avian and mammalian species , including sporadic but concerning cases in humans. There is growing concern that circulating H5N1 strains could lead to widespread human outbreaks . Research with highly pathogenic H5N1 viruses is restricted to Biosafety Level 3 (BSL-3) laboratories. Vesicular stomatitis virus (VSV)-based vaccine vectors expressing heterologous viral proteins from Ebola, SARS-CoV-2, Lassa virus, etc., have previously been shown to be safe and effective in animal models and human clinical trials . Here, we report the development of a recombinant VSV expressing the hemagglutinin (HA) and neuraminidase (NA) genes of H5N1 IAV (H5N1-VSV), which serves as a versatile platform to study various aspects of H5N1 IAV biology. H5N1-VSV replicated robustly to titers comparable to those of the full H5N1 virus in multiple cell lines. In mice , H5N1-VSV vaccination was safe, elicit...

Abstract Transmission routes of highly pathogenic H5N1 between cows or to humans remain unclear due to limited data from affected dairy farms. We performed extensive air, farm wastewater, and milk sampling on 14 H5N1 positive dairy farms across two different California regions. Virus was detected in the air in milking parlors and from exhaled breath of cows . Infectious H5N1 virus was detected in the air and water streams ; sequence analysis revealed viral variants on a farm in these locations. Longitudinal analysis of milk from the individual quarters of cows revealed a high prevalence of subclinical H5N1 positive cows and a heterogeneous distribution of infected quarters that maintained a consistent pattern over time. Our data highlight potential modes of H5N1 transmission on dairy farms. Competing Interest Statement SSL and LCM receive funds from Flu Lab and NIH. ASL receives funds from Flu Lab, NIH, and CDC. ASL receives consulting fees and research support from Roche, outside of t...

Abstract In 2024, highly pathogenic avian influenza A H5N1 caused outbreaks in wild birds, poultry, cows, and other mammals in the United States with 61 human cases also reported by the CDC. Detection of influenza A H5 RNA in wastewater has been previously reported in sewersheds in Texas and North Carolina with nearby impacted dairy herds following the emergence of H5N1 in dairy cows. Here, we conduct retrospective testing of total influenza A and H5 hemagglutinin genes in wastewater as well as present and apply new assays for detection of H1 and H3 genes across a respiratory virus season in an urban California sewershed from September 2023 to May 2024. Total influenza A, H1, and H3 were regularly detected, while H5 was first detected in March . We developed a model that uses Monte Carlo simulations and previously published parameters to estimate the numbers of infected people, poultry, wild birds, or liters of H5-contaminated milk required to result in measured H5 concentrations in wa...

  Key points   -- At the present time , based on available information, FAO-WHO-WOAH assess the global public health risk of influenza A(H5) viruses to be low , while the risk of infection for occupationally or frequently exposed (e.g., with backyard poultry) persons is low to moderate depending on the risk mitigation and hygiene measures in place and the local avian influenza epidemiological situation.  -- Transmission between animals continues to occur and, to date, a growing yet still limited number of human infections are being reported. Although additional human infections associated with exposure to infected animals or contaminated environments are expected to occur, the overall public health impact of such infections at a global level , at the present time, is considered minor . The assessment could change if and when additional epidemiological or virological information becomes available.  -- This risk assessment from FAO, WHO and WOAH updates the assessment ...

Abstract The emergence of highly pathogenic avian influenza (HPAI) H5N1 (clade 2.3.4.4b, genotype B3.13) in dairy cattle presents substantial challenges to the agricultural sector and public health. Mechanistic studies of infection and transmission in cattle have proven difficult due to animal handling restrictions as well as limited availability of established cell culture models. Primary Bovine Embryonic Fibroblasts (BeEFs) were collected from a Montana cow and are investigated here as a model to study influenza A virus (IAV) infection dynamics . We compared sialylation profiles, infectious virus production, viral replication, and plaque morphology in both BeEFs and chicken DF-1 cells following infection with the bovine HPAI H5N1 and an earlier 2.3.4.4b genotype (B1.1) isolated in 2022. The data presented here show increased viral fitness of the bovine origin HPAI H5N1 strains across multiple species and bovine susceptibility to human seasonal IAV. This study highlights the ability o...

Abstract Highly pathogenic avian influenza (HPAI) H5N1 virus has been associated with severe mastitis in dairy cows , leading to decreased milk production . Here we investigated the impact of H5N1 virus infection in health and production parameters in an affected dairy herd in Ohio . Clinical disease , which lasted for about three weeks , was recorded in 20.0% (777/3876) of the adult cows. Milk losses of ~900 kg per cow were recorded in affected cows during a 60 day-post-outbreak period . Seroprevalence was 89.4% (570/637) in the herd, with 76.1% (485/637) of seropositive animals being subclinically infected . Clinically affected cows presented an increased risk of death (6 times) and of premature herd removal (3.6 times) when compared to non-clinical cows. Economic losses due to decreased milk production, mortality, and early herd removal were estimated at $950 per clinically affected cow for a total cost of ~$737,500 for the herd during the observation period. Our results demonstrate...

ABSTRACT In recent years, high pathogenicity avian influenza viruses (HPAIVs) have spread among wild, captive, and domestic birds, as well as mammals . Beyond the resulting economic and ecological losses , spillover into mammals has raised concerns about a potential pandemic . Viral tropism refers to the spectrum of host species, organs, and cells susceptible and permissive to viral infection . It is a potent driver of infection dynamics and shedding patterns, which presents important variations both between and within hosts: in poultry, HPAIV leads to systemic endothelial infection in domestic chickens , whereas neurological and selective epithelial infections are observed in domestic ducks . In mammals , infection can result in respiratory and neurological disease , but the recent outbreaks in domestic dairy cows highlighted a unique and remarkable adaptation to the mammary gland prone to viral shedding in milk. The present review explores viral tropism of HPAIV across recent spillov...

{Excerpt} 1. Background   A risk assessment exercise using the Tool for Influenza Pandemic Risk Assessment ( TIPRA ) was conducted for Clade 2.3.4.4b A(H5N1) genotype B3.13 virus in Aug – Oct 2024 .   From late 2023, unexplained illnesses were reported in dairy cattle across multiple states in the United States .  Following those events , detection of avian influenza A(H5N1) in dairy cattle from Texas was confirmed on 25 March 2024.  As of 9 December 2024 , the virus has been detected in 720 dairy cattle herds across 15 states, according to the United States Department of Agriculture (USDA), with a total of 47 human infections from poultry and dairy cattle sources, excluding 11 human cases from Washington state , that might belong to genotype D1.1, also reported.  Wild and domestic animals (e.g., cats, raccoons) near infected herds have been affected as well.  The outbreak remains uncontrolled .  Due to the unprecedented spread of the virus among ...