Highly pathogenic avian influenza (HPAI) is a transboundary and zoonotic viral disease affecting poultry and wild birds in many countries worldwide. Globally, HPAI outbreaks have led to the death or culling of hundreds of millions of birds over the past two decades and have caused nearly 1000 confirmed human H5N1 infections, with a case fatality rate of approximately 50%. Asia and Europe remain among the most affected regions, with recurrent outbreaks linked to intensive poultry production, live bird markets, and migratory bird pathways. In Nepal, HPAI has been reported since 2009, with more than 320 outbreaks recorded and over 2.7 million birds lost, alongside one confirmed human fatality. Control measures rely largely on stamping out, movement restrictions, and surveillance; however, gaps in farm-level biosecurity, informal cross-border poultry trade, and limited vaccination use continue to sustain vulnerability. Strengthened multisectoral coordination under a One Health framework, integrating veterinary and public health surveillance, molecular monitoring, community awareness, and risk-based biosecurity enforcement, is essential to reduce the impact of HPAI and mitigate future zoonotic and pandemic risks.

The dairy sector in Nepal has a high potential for growth, but unfortunately, its progress is impeded by diseases such as mastitis. The purpose of the current investigation was to identify the predominant bacterial species in mastitis-affected cattle milk and to ascertain the most efficacious antibacterial treatment against the bacterial isolates. The study was conducted from 17th August 2022 to 12th February 2023 in lactating cattle of the Kirtipur municipality and Chandragiri municipality of Kathmandu district, Nepal. A total of 640 milk samples from 160 cattle were collected randomly from cattle farms of Kirtipur and Chandragiri municipality of Kathmandu district. Initial screening for mastitis was conducted on collection sites by the California Mastitis Test (CMT) and further bacterial isolation and identification were carried out at the Central Veterinary Laboratory using standard bacterial techniques and the antibiogram were evaluated based on CLSI guidelines. Among 640 samples collected from 160 cattle, 3.91% (25/640) tested CMT-positive at the individual quarter level, whereas the prevalence at the cow level was 15.62% (25/160). The major bacterial species isolated, in subsequent culture of CMT positive samples, were Staphylococcus spp. (10, 40%), E. coli (6, 24%), Streptococcus spp. (5, 20%), Enterococcus spp. (2, 8%), Bacillus spp. (1, 4%), and Klebsiella spp. (1, 4%).. Antibiogram study reveals that major gram-positive bacteria, Staphylococcus spp., Streptococcus spp., Enterococcus spp., and Bacillus spp. were highly sensitive to Ampicillin, Florfenicol, and Gentamicin while the major gram-negative bacteria, E. coli, and Klebsiella spp. were highly sensitive to Florfenicol, Gentamicin, Trimethoprim/Sulfamethoxazole and Ceftriaxone. Hence, early diagnosis and careful monitoring of antimicrobial susceptibility before therapeutic intervention are essential to prevent high economic losses due to mastitis and to mitigate the potential risk of antimicrobial resistance in the livestock population.