Prevalence and Antimicrobial Resistance Patterns of Salmonella in Asymptomatic Horses in Eastern Spain: A One Health Perspective

<b>(1) Background:</b><i>Salmonella</i> are zoonotic pathogens, and rising antimicrobial resistance (AMR) amplifies their public health impact. Asymptomatic horses can act as reservoirs, contributing to environmental contamination and interspecies transmission. This study aimed to estimate the prevalence of <i>Salmonella</i> and characterize AMR patterns in healthy horses from eastern Spain. <b>(2) Methods:</b> Faecal samples from 95 asymptomatic horses were collected once daily over five consecutive days (475 samples in total) and processed under for <i>Salmonella</i> detection. Epidemiological information was obtained through owner questionnaires, and associations with <i>Salmonella</i> shedding were analyzed using generalized linear models. Antimicrobial susceptibility was assessed by minimum inhibitory concentration assays following the European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. <b>(3) Results:</b><i>Salmonella</i> was detected in 25.3% of horses (24/95), with S. Enteritidis, S. Johannesburg, and S. Virchow as the most frequent serotypes. A significant association was observed between proximity of manure storage and bacterial detection (<i>p</i> < 0.001). Among 24 isolates of <i>Salmonella</i>, 88.9% were resistant to at least one antimicrobial, and 50% exhibited multidrug resistance. The highest resistance rates were against sulfamethoxazole and gentamicin, followed by ciprofloxacin and tigecycline. <b>(4) Conclusions:</b> Healthy horses can act as silent carriers of multidrug-resistant <i>Salmonella</i>, highlighting the need for surveillance, strengthened biosecurity, and prudent antimicrobial use within a One Health framework.