Dairy Cows’ Udder Pathogens and Occurrence of Virulence Factors in Staphylococci

SIMPLE SUMMARY: Dairy farmers and those involved in primary milk production face the challenge of maintaining the health of their animals. Although the level of technological support during milking has increased and appropriate veterinary measures are available, mastitis remains a major health problem in dairy cows as well as a major threat to dairy farm profitability. The results of this study indicate that, in addition to the major udder pathogens (S. aureus, S. uberis and S. agalactiae) causing dairy mastitis, non-aureus staphylococci (NAS) are a major risk to dairy cows during early lactation. NAS, such as S. chromogenes, S. warneri and S. xylosus, which were isolated from animals with clinical mastitis characterized by mild, moderate, or severe symptoms and persistent infections, had the highest representation of virulence factors (production of hemolysis, gelatinase and biofilm; the ability to hydrolyze DNA; resistance to antibiotics) in comparison to less virulent strains. In addition, isolates S. aureus and NAS showed resistance to one or more antimicrobials in 77.2%, and in 16 (11.7%) isolates, multi-drug resistance to three or more antimicrobial classes was confirmed. Due to the high resistance to β-lactam-antibiotics in two isolates of S. aureus and two species of NAS, the presence of a methicillin-resistant gene, mecA, was confirmed, which potentially poses serious complications for the treatment of dairy cows and a serious health risk to milk consumers. ABSTRACT: This study investigated 960 Slovak and Czech spotted cattle from four different conventional (non-organic) dairy herds located in Eastern Slovakia and Czechia during early lactation (14–100 days after calving). Dairy cows were examined clinically; milk from fore-stripping of each udder quarter was subjected to sensory examination and assessed by the California mastitis test (CMT), and laboratory analyses of bacterial pathogens in milk, including virulence factors, were conducted. Positive CMT scores (1–3) for one or more quarters were detected in 271 (28.2%) of the examined animals. Out of 230 infected milk samples, representing 24.0% of all dairy cows, staphylococci (59.1% of positive findings) were the most commonly isolated organisms, followed by E. coli (11.3%), streptococci Str. uberis (9.1%) and Str. agalactiae (3.4%), and enterococci (6.1%). From 136 isolates of S. aureus (38 isolates) and non-aureus staphylococci (NAS; 98 isolates), virulence factors and their resistance to 14 antimicrobials were detected using the disk diffusion method, with PCR detection of the methicillin resistance gene, mecA. An increased incidence of clinical and chronic forms of mastitis has been reported in mastitic cows in which staphylococci, especially S. aureus and NAS (S. chromogenes, S. warneri, and S. xylosus), have been detected and compared to other isolated udder pathogens. From those species, S. aureus and isolates of NAS mentioned above showed multiple virulence factors that are more likely to hydrolyze DNA, hemolysis, produce gelatinase and biofilm, and have multi-drug resistance as compared to other less virulent staphylococci. Generally, the isolated staphylococci showed 77.2% resistance to one or more antimicrobials, in particular to aminoglycosides, β-lactams, macrolides, or cephalosporins. Isolates that showed the ability to form a biofilm were more resistant to more than one antimicrobial than isolates without biofilm production. Multi-drug resistance to three or more antimicrobial classes was recorded in 16 isolates (11.7%), and the presence of the mecA gene was also confirmed in two isolates of S. aureus and two species of NAS.