Abstract
Bovine mastitis is mainly caused by bacteria of the genus Staphylococcus spp., which possess different virulence factors, including the capacity for biofilm formation that provides enhanced protection against the action of immune system components and serves as a barrier against the penetration of antimicrobial agents. This study aimed to characterize 181 Staphylococcus spp. Strains—including Staphylococcus aureus and coagulase-negative staphylococci (CoNS) isolated from bovine subclinical mastitis in six Brazilian states—by molecular methods. RT-qPCR was used to verify the expression of genes of the ica operon—mainly responsible for biofilm formation—as well as bap and bhp. Chromosome similarity among the isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The icaA gene was detected in 79 (43.6%) isolates, icaB in 24 (13.2%), icaC in 57 (31.4%), and icaD in 127 (70.1%). The bap gene was identified in 66 (36.4%) isolates, while the bhp gene was found in nine (4.9%). RT-qPCR confirmed the expression of the icaA gene in 60 (75.9%) isolates, of icaB in six (25%), of icaC in 26 (45.6%), and of icaD in 80 (63%). Clonal typing of the isolates by PFGE permitted the identification of eight Staphylococcus aureus clusters that simultaneously included ≥3 strains, with a similarity of ≥80%. Regarding the other species studied, three clusters were observed for Staphylococcus chromogenes and four clusters for Staphylococcus epidermidis. Only one cluster each was identified for Staphylococcus saprophyticus and Staphylococcus simulans, while the other species did not form any cluster. With respect to MLST, ST126 and ST1 were the prevalent sequence types in S. aureus, while in S. epidermidis all sequence types were different. These results reveal strains with the same evolutionary origin as other isolates, which might cause infections in humans and animals, suggesting their ability to spread between these species.
Highlights
Inadequate handling of livestock can trigger inflammatory processes in the mammary gland, known as mastitis, the most prevalent and costly disease in the dairy industry
Detection of Genes Involved in Biofilm Formation
The presence of biofilm formation-related genes was investigated in the genomic DNA of all isolates of the study
Summary
Inadequate handling of livestock can trigger inflammatory processes in the mammary gland, known as mastitis, the most prevalent and costly disease in the dairy industry. Mastitis is caused by different microorganisms that directly influence the physicochemical characteristics, composition and cellularity of milk [1,2]. The economic consequences of both clinical and subclinical mastitis include losses due to the costs of treatment, lower milk production, changes in product quality, and culling [2]. Mastitis is a public health hazard since it can cause zoonoses and food poisoning [3,4]. Few studies report the prevalence of subclinical mastitis in Brazilian dairy herds [5]. Wide variation exists in the cow-level prevalence of this disease
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