Abstract
This study comprehensively explored the epidemiology, virulence, antibiotic resistance, and genomic features of ovine-derived Mannheimia haemolytica across three provinces in Northwest China (Gansu, Ningxia, and Shaanxi). Lung tissue samples were collected postmortem from sheep with respiratory disease (nine per province), cultured under sterile conditions, and screened by morphological, Gram staining, PCR, and NGS methods. Nine M. haemolytica strains were successfully isolated (isolation rate 33.33%) and identified as serotypes A1, A2, and A6, with Shaanxi exhibiting a significantly higher isolation rate of 66.67%. Genomic analysis revealed that each isolate each harbored over 100 virulence genes and displayed notable genetic diversity. In murine assays, intraperitoneal inoculation in mice caused severe breathing problems and death within 24 h; necropsy revealed lung congestion, splenic necrosis foci, and hepatic congestion. Antibiotic susceptibility testing showed sensitivity to ciprofloxacin, azithromycin, gentamicin, and levofloxacin, while resistance was observed for tiamulin and enrofloxacin. These findings indicate a high prevalence and strong pathogenicity of ovine M. haemolytica in Shaanxi, underscore the presence of key resistance traits, and provide a genomic and pathogenic reference to guide region-specific surveillance, therapeutic decisions, and vaccine-oriented control in small ruminant herds. Importantly, these results highlight the need for region-specific monitoring and judicious antibiotic use, which are essential to forestall the spread of resistant strains and to ensure sustainable disease management strategies.