Search Results (134)

Avian influenza viruses continue to pose significant zoonotic and pandemic threat globally, with low-pathogenic avian influenza A(H9N2) being of particular concern due to sustained circulation in poultry, adaptability, and repeated human spillover. This study investigated the detection and genetic characterization of influenza viruses at the animal–human interface in Ghana in 2022, using a nationwide cross-sectional One Health approach. Samples were collected from poultry, pigs, the environment, and animal handlers across backyard farms, commercial farms, and live bird markets. Laboratory testing was conducted using real-time RT-PCR, while statistical associations were assessed using chi-square and logistic regression. Whole-genome sequencing and phylogenetic analysis were performed on selected positive samples. Out of 4056 samples, 1516 were poultry samples. A(H9N2) was detected exclusively in poultry, with a prevalence of 5.67%. The Northern belt recorded the highest prevalence. Live bird markets had significantly higher odds of A(H9N2) detection compared with commercial farms (odds ratio: 15.37, p < 0.0001), while backyard farms had lower odds. Environmental samples were negative. Among animal handlers, one case each of A(H3N2) and SARS-CoV-2 was identified. Phylogenetic analysis demonstrated that Ghanaian strains belonged to clade G1 and possessed mammalian-adaptive markers. These findings highlight ongoing circulation in poultry and the need for sustained One Health surveillance. Full article

Highly pathogenic avian influenza (HPAI) A(H5N1) viruses of clade 2.3.4.4b have recently spread across the Americas, prompting intensified surveillance efforts in Brazil aimed at early detection in wild birds. As part of these efforts, we identified a low pathogenic avian influenza A(H11N2) virus in a white-rumped sandpiper (Calidris fuscicollis) sampled at Lagoa do Peixe National Park (PNLP) in southern Brazil. Whole-genome sequencing revealed that seven of the eight gene segments shared high nucleotide similarity (approximately 98.8%) with viruses previously detected in shorebirds from Delaware Bay, North America. In contrast, the PB1 segment showed high nucleotide similarity (approximately 99%) to the PB1 lineage associated with clade 2.3.4.4b A(H5N1) genotype B3.2 viruses circulating in the Americas. Phylogenetic, nucleotide identity, and molecular clock analyses indicated that this lineage shares a recent common ancestor with North American LPAI viruses and was subsequently detected in distinct viral genetic backgrounds. Although no HPAI virus was identified in this study, the presence of a PB1 segment related to H5N1-associated lineages suggests that genetic components linked to these viruses were circulating among low pathogenic avian influenza viruses in South America. These findings highlight the importance of continued surveillance in migratory bird populations to improve understanding of avian influenza virus diversity and support epidemiological monitoring. Full article

Highly pathogenic avian influenza (HPAI) H5N1 viruses of the goose/Guangdong (Gs/GD) lineage have driven a global panzootic since 2020, with clade 2.3.4.4b establishing sustained transmission in wild birds. In South America, early outbreaks were largely associated with the North American-derived B3.2 genotype, which showed limited diversification after its introduction. Here, we report the genomic characterization of eight H5N1 viruses detected in Uruguay during the reemergence of avian influenza in February–March 2026. Complete genomes were obtained from wild birds exhibiting neurological signs, predominantly Coscoroba coscoroba. All viruses belong to clade 2.3.4.4b but exhibit a reassortant genomic constellation distinct from B3.2. The HA, NA, and MP segments retain the Eurasian backbone, whereas internal genes derive from both South American and North American low-pathogenicity avian influenza lineages. PB2 variation distinguishes two closely related viral groups differing in PB2 origin, whereas the remaining genomic segments retain a shared background. Sequence variation in the neuraminidase gene reduced the sensitivity of a widely used N1-specific RT-qPCR assay, highlighting limitations of existing diagnostic tools during viral evolution. These findings confirm the presence of reassortant H5N1 viruses in Uruguay and, together with recent reports from Argentina and Brazil, support an emerging pattern of genomic diversification in southern South America. Full article

Wastewater surveillance emerged as a critical public health tool during the COVID-19 pandemic, enabling early detection of community-level pathogen circulation independent of clinical testing. Its ability to capture signals from both symptomatic and asymptomatic individuals highlighted the importance of optimizing sampling methodologies to improve sensitivity and reliability. A key question is whether the several-fold increase in SARS-CoV-2 detectability observed when using passive tampon swab sampling compared with paired grab samples also applies to other respiratory viruses, including influenza A (including its avian influenza H5N1 subtype), influenza B, and respiratory syncytial virus (RSV). We collected 24 h passive swab samples with same-day grab samples from Detroit sewersheds, concentrated and purified nucleic acids, and using RT-ddPCR, quantified respiratory syncytial virus, SARS-CoV-2, influenza A, influenza B, and H5N1 influenza A viruses using markers RSV, SC2, InfA, InfB, and H5, respectively. Samples testing positive for H5 (marker for H5N1 influenza A) were further analyzed by targeted PCR and amplicon sequencing. Across three sites, median 24 h swab:grab ratios of virus copies were 7.0 for RSV, 9.2 for SC2, 9.9 for InfA, and 3.6 for InfB. A 239 bp hemagglutinin sequence from a sample with a strong H5 signal (795 copies/10 mL) had 100% identity to avian influenza viruses from Canada geese. Twenty-four-hour swab sampling greatly improves viral detectability across diverse targets and enabled the first confirmed detection of H5 in Detroit wastewater. Combined with magnetic bead purification, the overall sensitivity gain over conventional PEG-NaCl-Qiagen methods is approximately 36-fold, enabling earlier warning of community pathogens than grab samples. By integrating 24 hour passive swab sampling with high-efficiency nucleic acid purification, we expand the sensitivity of wastewater surveillance to enable detection and confirmation of low-abundance pathogens like avian influenza (H5). Full article

Avian influenza (AI) continues to threaten global poultry production, with accumulating evidence suggesting that certain commercial layer lines may exhibit increased susceptibility under specific experimental conditions compared with broiler chickens. This narrative review synthesizes published experimental infection studies identified through a comprehensive PubMed search, focusing on low pathogenic H9N2 and highly pathogenic H5N1, H5N2, H7N7, and H7N9 viruses. Although bird age and production stage varied across studies, consistent disparities in immune regulation and viral replication dynamics have been reported. We critically evaluate host determinants underlying these differences—including microRNAs, major histocompatibility complex polymorphisms, sialic acid receptor distribution, gut microbiota, and hormonal influences—and integrate findings across viral subtypes and pathogenicity classes to inform breed-tailored vaccination, nutritional, and therapeutic strategies. Full article

Background/Objectives: Biosecurity and stamping out are key control measures against H5 high pathogenicity avian influenza (HPAI) outbreaks. Vaccination in poultry is an additional tool to reduce disease risk and facilitate timely containment. This study aimed to establish a candidate vaccine strain against H5 HPAI in Asia and validate its protective efficacy. Methods: Based on genetic and antigenic analyses, a representative HPAI virus, A/duck/Vietnam/HU16-DD3/2023 (H5N1), collected in northern Vietnam, was selected to generate a candidate vaccine strain, rgPR8/VN23HA∆KRRK-NA (rgPR8/VN23; H5N1), using reverse genetics, followed by formulation of an inactivated oil-adjuvanted vaccine. Vaccine efficacy was evaluated by measuring humoral antibody responses after intramuscular vaccination and by assessing mortality and virus recovery following intranasal challenge with a clade 2.3.4.4b virus, A/Ezo red fox/Hokkaido/1/2022 (H5N1). Results were compared with those obtained using an antigenically homologous vaccine to the challenge strain and a Japanese stockpiled vaccine. Results: All vaccinated juvenile chickens developed sufficient immunity to survive the challenge at 21 days post-vaccination. The rgPR8/VN23 (H5N1) and homologous vaccines markedly reduced virus recovery, suggesting near-sterile protection, whereas low-titer viruses were transiently detected in chickens vaccinated with the stockpiled vaccine. The rgPR8/VN23 (H5N1) vaccine conferred clinical protection in juvenile chickens as early as 8 days post-vaccination. A single dose of the rgPR8/VN23 (H5N1) vaccine provided incomplete protection in laying hens, whereas a double-volume regimen improved protective efficacy. Conclusions: The rgPR8/VN23 (H5N1) vaccine conferred strong immunity to juvenile chickens; however, a refined vaccination strategy may be required to achieve complete protection in laying hens. Full article

Highly pathogenic influenza A viruses of the H5 subtype continue to diversify worldwide through mutation and genetic reassortment, generating novel variants with unpredictable consequences under the One Health approach. Between 2024 and 2025, five outbreaks of avian influenza A viruses were detected in backyard poultry across Michoacán, Estado de México, and Ciudad de México. We conducted molecular and genetic characterization of five highly pathogenic H5N2 viruses isolated from these events. All cases tested positive for influenza A virus and the H5 hemagglutinin, exhibiting high pathogenicity with intravenous pathogenicity index values ranging from 2.88 to 3.0. Whole-genome sequencing revealed novel reassortants containing hemagglutinin from Eurasian H5N1 clade 2.3.4.4b and neuraminidase from the endemic Mexican H5N2 lineage. The viral genome of the isolate from Michoacán contained six segments derived from Eurasian H5N1 viruses introduced into North America in 2021–2022, while nucleoprotein and neuraminidase originated from Mexican H5N2 viruses. In contrast, viruses from Estado de México and Ciudad de México contained five H5N1-derived segments and incorporated polymerase basic protein 1, nucleoprotein, and neuraminidase from low-pathogenic H5N2 viruses circulating in 2024. Phylogenetic analyses confirmed the emergence of a distinct H5N2 Mexican sublineage, providing evidence of active viral reassortment and local evolutionary processes in Mexico. Full article

Retinoic acid-inducible gene I (RIG-I) is a crucial pattern recognition receptor for detecting viral RNA and initiating an immune response against influenza A viruses (IAVs). The activation of RIG-I in mammalian cells requires ubiquitination by two E3 ubiquitin ligases: TRIM25 and RIPLET. Using dual luciferase assays, we demonstrate that duck RIPLET enhances the activation of RIG-I, driving the IFN-β promoter activity in chicken DF-1 fibroblasts. qPCR analyses show that the co-expression of duck RIG-I and RIPLET significantly upregulates key immune genes and reduces viral RNA transcripts in DF-1 cells challenged with low pathogenic avian influenza (LPAI) H6N2. Co-immunoprecipitation and confocal microscopy studies suggest the interaction and confirm the colocalization of duck RIG-I and RIPLET in the cytoplasm. Further, we show that the non-structural protein 1 (NS1) of IAV, known for its role in immune evasion, suppression, and pathogenicity, from five different strains of IAV (PR8, BC500, CA431, D4AT, and VN1203) can all inhibit duck RIPLET activation of RIG-I, with NS1 from avian strains showing the greatest decrease in IFN-β promoter activity in chicken DF-1 cells. Overall, our research provides valuable insight into the E3 ubiquitin ligases required for RIG-I activation and susceptibility of this pathway to IAV interference across species. Full article

Avian influenza viruses are predominantly associated with waterfowl and shorebirds, and are rarely detected in other avian hosts in nature. In 2021, an H1N1 virus was isolated from a Fieldfare Turdus pilaris in Zaporizhzhia Oblast, Ukraine. A phylogenetic analysis revealed that all eight gene segments belonged to the Eurasian low-pathogenic avian influenza lineages. The highest nucleotide identity of the HA gene was observed with viruses detected in Georgia, Sweden, and Ukraine (99.11%), while the NA gene showed the greatest identity to viruses from Western Europe (99.14–99.57%). Genetic analysis of the HA cleavage site showed a sequence (PSIQSR↓GLF) that contained a single basic amino acid. No deletions were detected in the stalk region of NA gene, and no specific mutations in PB2 protein were found. However, several amino acid substitutions were identified in the HA gene (D204E, S207T, and D239G) that may affect the binding affinity to specific antibodies. The occurrence of this virus in a wild, seemingly healthy thrush indicate that additional surveillance in poorly studied ecological groups such as Passeriformes is warranted. Full article

The H9N2 avian influenza virus (AIV) is difficult to prevent and control because of its low pathogenicity and frequent mutation. In a previous study, the HA (hemagglutinin) protein of H9N2 was expressed in a rice endosperm reactor and prepared into a subunit vaccine to immunize chickens and mice, both of which exhibited a good immunity effect. The results of the intermediate tests of the transgenic strains (AIV-1 and AIV-3) showed that the HA gene can be stably expressed. Agronomic traits, such as plant height and number of grains, were significantly optimized in the transgenic strains. Moreover, no exogenous HA genes were found in the leaves of the weeds, and it was initially determined that there was no risk of gene drift. This study provides key technical support for the commercialization of plant subunit vaccines for avian influenza viruses. Full article

The detection of H5N1 highly pathogenic avian influenza virus (HPAIV) in lactating dairy cattle in the United States, with high viral titers in raw milk, has raised concerns about zoonotic transmission through unpasteurized milk and dairy products. While viral inactivation during pasteurization is documented, data on persistence in raw-milk cheeses remain limited. This study evaluated the survival of avian influenza viruses (AIVs), both low pathogenic (LPAIV, H1N1) and highly pathogenic (HPAIV, H5N1), during the production and ripening of Grana-type hard cheeses from raw cow’s milk. Experimental cheesemaking was carried out with milk artificially contaminated with A/duck/Italy/281904-2/06 (H1N1; 10^7.75 EID₅₀/mL) or A/duck/Italy/326224-2/22 (H5N1 clade 2.3.4.4b; 10^6.75 EID₅₀/mL). Cheeses were manufactured under Parmigiano-Reggiano standards and ripened 30 days at 5–6 °C. Viral detection in finished cheeses was performed using inoculation in specific-pathogen-free embryonated chicken eggs (SPF-ECEs), hemagglutination (HA) assay, and monoclonal antibody-based ELISA. No infectious virus was detected in cheese samples after two blind passages in SPF-ECEs. Both HA and ELISA tests were negative, indicating complete viral inactivation. These results demonstrate that Grana-type cheese processing, including cooking, acidification, and ripening, effectively inactivates LPAIV and HPAIV. Findings support the microbiological safety of raw-milk hard cheeses regarding AIV, contributing to risk assessment and food safety policies. Full article

The H3N8 low pathogenic avian influenza virus (LPAIV) exhibits broad host tropism, infecting diverse avian and mammalian species, raising concerns about its zoonotic potential. Following the emergence of human infections with H3N8 LPAIV in China, including a fatal case, we investigated the epidemiological and virological characteristics of this virus in Guangdong Province. In 2022, a serological survey revealed H3N8 seroprevalence rates of 10.85% in farmed chickens and 7.97% in ducks. We isolated three H3N8 viruses, designated as A/chicken/Qingyuan/22/2022 (H3N8); A/chicken/Qingyuan/31/2022 (H3N8); and A/chicken/Qingyuan/15/2022 (H3N8), and found that these chicken isolates, like the human isolate A/Changsha/1000/2022, share the same E190 residue. This residue can synergize with sites such as Q226 and G228 to enhance binding affinity for SAα-2,6-Gal. Additionally, they harbor the three amino acid residues N193, W222, and S227. Among these, N193 has the potential to form hydrogen bonds with α2-6-linked glycans, while W222 and S227 may alter the conformational flexibility of the 220-loop. These two effects collectively endow the H3N8 isolates with dual receptor-binding properties. These findings suggest a shift in receptor specificity, potentially facilitating viral adaptation to mammalian hosts. Characterization of viral genome detection dynamics, and histopathology in animal models further elucidated the viral infection dynamics. Our study provides critical insights into the evolutionary trajectory and zoonotic potential of the H3N8 LPAIV. Full article

The H6N6 avian influenza virus has expanded its host range from birds to mammals. Some strains can now bind to human-like receptors, raising concerns about human infection. Although H6N6 is a low-pathogenic avian influenza virus (LPAIV), it is unclear whether it triggers pyroptosis in human lungs, a process linked to cytokine storms in infections like H7N9. Here, we found that the chicken-origin H6N6 LPAIV can effectively replicate in and infect human alveolar macrophages and their M1 macrophages. Viral infection of M1 macrophages upregulated the mRNA levels of NLRP3, caspase-1, and Gasdermin D (GSDMD). Subsequently, caspase-1 was activated and cleaved GSDMD protein into its N-terminal fragment (GSDMD-N), which formed pores in the cell membrane and triggered the release of IL-1β and IL-18. Further analysis demonstrated that inhibition of the NLRP3/Caspase-1/GSDMD pathway by specific inhibitors attenuated pyroptosis in infected M1 macrophages. In summary, our study revealed that H6N6 virus infection induces M1 macrophage pyroptosis via the NLRP3/caspase-1/GSDMD pathway. Notably, M1 macrophages inherently produce pro-inflammatory cytokines; their pyroptosis, accompanied by the release of IL-1β and IL-18, can amplify inflammation and potentially trigger a cytokine storm in the lungs. These findings reveal novel pathogenic mechanisms and potential therapeutic targets for avian influenza viruses. Full article

Rapid and accurate diagnostics for influenza viruses are essential for preventing future epidemics. Surface-enhanced Raman spectroscopy (SERS) presents a promising alternative to conventional techniques, offering a rapid, cost-effective, and highly sensitive platform for influenza virus detection. It is a highly sensitive analytical technique that enables the detection of minute chemical substances through significant signal enhancement. It operates by illuminating a sample with a laser and analyzing the scattered light to generate a unique molecular Raman spectrum. The sensitivity of SERS is derived from its use of metal nanoparticles, which amplify the weak Raman signals, making it particularly effective for detecting low-concentration targets such as viruses. Avian influenza (AI) is a major threat to domestic poultry, leading to large-scale culling during outbreaks. It leads to economic losses globally and can also infect pigs and humans, potentially causing a pandemic. Migratory birds spread various strains, leading to the development of highly pathogenic viruses. Viral monitoring is crucial for prevention strategies and understanding the virus evolution. This review outlines the challenges in detecting AI virus in chickens and critically assesses the established and emerging diagnostic technologies, with a specific focus on the factors influencing detection and recent advances in SERS-based AI detection. Ultimately, this review aims to provide insights that will assist the influenza research community in developing novel strategies for monitoring and preventing AI outbreaks in chickens and mitigating zoonotic transmission. Full article

H7 high-pathogenicity avian influenza (HPAI) virus outbreaks can cause high rates of morbidity and mortality in poultry flocks, leading to devastating impacts on poultry industries. In December 2024, an HPAI virus was detected on a poultry farm in New Zealand, being the first time a case of HPAI was reported in the country. Whole-genome sequencing, subtyping, phylogenetic, and mutation analyses were performed to characterize the virus. Results indicated a novel high-pathogenicity H7N6 avian influenza virus arose through a reassortment event between endemic low-pathogenicity H4N6 and H7 viruses, followed by two mutations at the H7 gene cleavage site. Mutation analysis suggests the novel H7N6 virus exhibits increased risk of host specificity shift, but further work is required to fully understand the functional impacts of the detected mutational events. In this instance, a timely biosecurity response was effective in eliminating the virus and preventing its transmission to secondary poultry flocks in New Zealand. However, the event underscores the critical importance of continued surveillance of commercial poultry and other potential avian carriers to facilitate early detection of low-pathogenicity avian influenza viruses, which may undergo reassortment or de novo mutation into high-pathogenicity variants. Full article