A One Health approach metagenomic study on the antimicrobial resistance traits of canine saliva

According to the One Health concept, physical proximity among pets and their owners facilitates the spread of various bacteria. Interspecies bacterial transfer media include canine saliva that can be transmitted by licking and biting. Antimicrobial resistance genes (ARGs) are the natural constituents of the bacterial genome. However, human activity can increase the prominence of ARGs. To assess factors that may affect the resistome of the canine oral cavity, the shotgun metagenomic sequencing dataset of 1830 canine saliva samples was bioinformatically analyzed and supplemented with survey results of the physical and behavioral traits of the dogs. Bacteriome and resistome associated with the canine saliva samples were described throughout the analysis. Moreover, the subset of higher public health ARGs and ESKAPE pathogen-related (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) higher public health ARGs were also collected. Further on, the set and subsets of ARGs were studied considering the surveyed traits of the sampled dogs. Overall, 318 ARG types reached sufficiently high detection rates. These ARGs can affect 31 antibiotic drug classes by various resistance mechanisms. ARGs against tetracyclines and cephalosporins appeared in the highest number of samples. However, surprisingly, another CIA group, peptides were represented by ARGs in the third-highest number of samples. Considering further ARG appearance rates in the samples, Critically Important Antimicrobials (CIAs, WHO), such as aminoglycosides, fluoroquinolones, or macrolides, were among the most frequently affected drug classes by higher public health risk ARGs and ESKAPE pathogen related higher public health risk ARGs. Bacteria in the saliva of white and diluted (merle, gray) color dogs and dogs characterized with decreased activity and decreased aggression more often harbored ARGs. Reduced playfulness could have been specifically associated with higher public health risk ARG presence. Even though the oral microbiome of the owners is unknown, One Health and public health implications of the close human-pet bonds and factors potentially underlying the rise in salivary ARG numbers should be considered, mostly in the light of the presence of ARGs affecting critically important drugs for human medicine. ### Competing Interest Statement The authors have declared no competing interest.