Abstract
INTRODUCTION: Brucella canis is a zoonotic pathogen that infects both dogs and humans, yet its evolutionary and phylogenetic characteristics are poorly understood. METHODS: Here, we comprehensively characterized an isolated strain of B. canis through integrated bacteriological, comparative genomic, and whole-genome sequencing-based core genome single-nucleotide polymorphism (WGS-cgSNP) analyses. RESULTS: B. canis YN20042 was isolated from a febrile patient (38 °C) with sweating and fatigue. The culture exhibited rough, grayish white, sticky, and opaque colonies. The isolate was identified as Brucella strain by a BCSP-31 polymerase chain reaction (PCR) assay, which yielded an amplicon of the expected 223-bp size, and was classified as a B. canis strain by conventional biotyping. The patient reported frequent contact with dogs and livestock. The strain showed a 99.99% average nucleotide identity to the B. canis reference strain ATCC 23365 (GCA_000018525.1). An in silico multilocus sequence typing (MLST) analysis showed that the strain belonged to sequence type 21, which was consistent with its classification within B. canis. The genome of strain YN20042 exhibited strong synteny with the reference strain and showed no detectable structural variations. It harbored 12 predicted virulence factors encompassing 71 associated genes, although it notably lacked the wbpL gene but contained a Brucella suis mprF gene. A further analysis identified predicted mutations in key virulence genes (eryA, pagN, bmaC, cfa1, and cfa2) and predicted multiple horizontally acquired genes, collectively suggesting a complex evolutionary trajectory involving both gene variants and potential recombination events. A WGS-SNP analysis revealed that YN20042 clustered closely with strains isolated from Zhejiang and Beijing, indicating a high degree of genetic relatedness. CONCLUSION: The first isolation of B. canis in the region expands the local spectrum of pathogenic Brucella and highlights the substantial infection risk for individuals with close dog and livestock contact. Enhanced surveillance, targeted screening of high-risk populations, and public health education are necessary to mitigate the risk of B. canis transmission.