This study aimed to investigate the diversity of milk isolates of Streptococcus uberis from Swedish dairy cows with mastitis, focusing on antibiotic resistance and virulence genes. We analyzed 115 S. uberis isolates using whole genome sequencing revealing a high level of diversity. Within the same farms, we identified both indistinguishable strains with identical sequence types (ST), and distinct isolates belonging to different ST types. This suggests both clonal and non-clonal spread of the bacterium, although primarily non-clonal. We found small clusters of two to eight closely related isolates both within and between farms. Differences in penicillin susceptibility were observed, probably linked to specific variants of penicillin-binding proteins. Few isolates were resistant to antibiotics, and few resistance genes were detected. In most cases, only one or two resistance genes were present, and only one isolate was multi-drug resistant. Two isolates had resistance genes against tetracyclines, a tet(M) and a tet(O) gene, two had a resistance gene against lincosamides, an lnu(C) and an lnu(D) gene, while a single isolate had an erm(B) gene conferring resistance to both macrolides and lincosamides. A single isolate carried a mef(A) gene, which confers resistance to macrolides via an efflux pump mechanism. However, we found aminoglycoside genes in 10 isolates, all 10 had the ant(6)-Ia gene, and one in addition aph(3')-IIIa, and a spectinomycin resistance gene, spw, in eight isolates. Finally, one isolate carried a streptothricin resistance gene, sat4. The genes sat4 and spw have apparently not previously been reported in S. uberis. Interestingly, isolates with elevated MIC to penicillin also significantly more often carried other resistance factors. Most isolates carried several virulence genes, including genes for capsule formation, adhesion to host cells or extracellular matrix proteins, and acquisition of essential nutritional factors, such as amino acids, iron and manganese.
Keywords: Antibiotic resistance genes, virulence; Dairy cows; Mastitis; Sweden; WGS; Whole genome sequencing.
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