Small ruminant mastitis is a serious problem, mainly caused by Staphylococcus spp. Different virulence factors affect mastitis pathogenesis. The aim of this study was to investigate virulence factors genes for biofilm production and antimicrobial resistance to β-lactams and tetracyclines in 137 staphylococcal isolates from goats (86) and sheep (51). The presence of coa, nuc, bap, icaA, icaD, blaZ, mecA, mecC, tetK, and tetM genes was investigated. The nuc gene was detected in all S. aureus isolates and in some coagulase-negative staphylococci (CNS). None of the S. aureus isolates carried the bap gene, while 8 out of 18 CNS harbored this gene. The icaA gene was detected in S. aureus and S. warneri, while icaD only in S. aureus. None of the isolates carrying the bap gene harbored the ica genes. None of the biofilm-associated genes were detected in 14 isolates (six S. aureus and eight CNS). An association was found between Staphylococcus species and resistance to some antibiotics and between antimicrobial resistance and animal species. Nine penicillin-susceptible isolates exhibited the blaZ gene, questioning the reliability of susceptibility testing. Most S. aureus isolates were susceptible to tetracycline, and no cefazolin or gentamycin resistance was detected. These should replace other currently used antimicrobials.

Antimicrobial resistance is a serious problem for the control of infections and infectious diseases. Propolis is a substance produced by honeybees with antimicrobial and antibiofilm properties. To consider propolis as an alternative to the use of antimicrobials for infection control, we assessed its antimicrobial and antibiofilm activities. To assess propolis for topical medical use, toxicological studies were also performed. A Portuguese 70% propolis ethanolic extract was chemically evaluated and studied for antimicrobial activity on staphylococcal field isolates (n = 137) and antibiofilm action (n = 45). Cell toxicological assessment was performed using keratinocytes and fibroblasts. Pinobanksin, chrysin, acacetin, apigenin, pinocembrin, and kaempferol-dimethyl-ether were detected. All 137 isolates were susceptible to 6.68 mg/mL or lower propolis concentration (80% isolates were susceptible to <1 mg/mL). The mean percentage of biofilm inhibition was 71%, and biofilm disruption was 88.5%. Propolis (<1 mg/mL) was well-tolerated by fibroblasts and moderately tolerated by keratinocytes. The combined antimicrobial and antibiofilm effect of propolis, together with its low toxicity to connective tissue and epithelial cells, suggests a good applicability for topical antibacterial treatment. Therefore, propolis seems to be a good alternative to antimicrobials for the treatment of infections with Staphylococcus spp. that deserves to be evaluated in vivo.