The aim of this study was to compare the sensitivity to selected antibiotics in staphylococci isolated from goat (n = 60), sheep (n = 60) and cow’s milk (n = 120). The individual milk samples were inoculated onto Blood agar cultivated at 36 °C/24 h.The isolated species of staphylococci were identified using biochemical tests, namely STAPHYtest and identification program TNW pro 6.5. We examined the sensitivity of strains to the spectrum of antibiotics, as follows: vancomycin (VA), amoxicillin/clavulanic acid (AMC), penicillin (P), rifampicin (RD), oxacillin (OX), tetracycline (TE), erythromycin (E), chloramphenicol (C), clindamycin (DA), gentamicin (CN), ciprofloxacin (CIP), teicoplanin (TEC), cefoxitin (FOX) and novobiocin (NOV). Altogether, 97 staphylococcal isolates were obtained; 70 from cow’s milk, 11 from goat’s milk and 16 from sheep’ milk. Staphylococcus aureus was the most frequent species in milk of all animal origin tested, was detected in 54 (45%) cow’s milk, 10 (17%) goat’s and 15 (25%) sheep’ milk samples. S. lentus was identified only in goat’s and sheep’ milk whereas in cow’s milk there were representation of staphylococcal species as follows: S. haemolyticus (n = 7), S. chromogenes (n = 2), S. warneri (n = 2), S. xylosus (n = 2), S. epidermidis (n = 2) and unclassified staphylococci (n = 1). The results of S. aureus sensitivity are similar for all tested antibiotics and for all monitored milk: No resistance to vancomycin, rifampicin, chloramphenicol and teicoplanin was recorded in obtained S. aureus isolates whereas the resistance to ciprofloxacin was found out most often.

ABSTRACT:The aim of this study was to report the detection of methicillin-resistant Staphylococcus aureus (MRSA) strains at a veterinary university goat breeding farm and their characteristics. A total of 278 samples collected from animals, milk, environment and farm personnel between June 2006 and March 2008 were examined. The identification of S. aureus isolates was performed by a species specific PCR assay. All detected isolates were tested for resistance to oxacillin and other antimicrobials by phenotypic methods and for the mecA gene by PCR method. Eight MRSA were detected in this study. Five of them originated from goat's milk and three were recovered from one human carrier of the farm personnel. All obtained MRSA isolates were clonally consistent and were characterized as: SCCmec type IV, spa type t064, seb positive and for genes encoding TSST-1, PVL and exfoliative toxins A and B negative.

This work is focused on the monitoring of Staphylococcus aureus prevalence in raw milk and milk filters, its antibiotic resistance and detection of methicillin resistant Staphylococcus aureus (MRSA). Samples of raw cow´s milk and milk filters were collected in the period from 2012 till 2014, from 50 dairy farms in the Czech Republic. The total of 261 samples (164 samples of raw milk and 97 milk filters) were cultivated on Baird-Parker agar. Both the typical and atypical colonies were examined by plasmacoagulase test and PCR method was used for detection of species specific fragment SA442 and mecA gene. Standard disk diffusion method was used to determinate resistance to antimicrobial agents. The bacterium Staphylococcus aureus was detected on 25 farms (50%). The antimicrobial resistance showed differences between the farms. Total of 58 samples were positive for Staphylococcus aureus, of which were 37 (14.2%) isolated from raw milk samples and 21 (8.1%) from milk filters. From these samples we isolated 62 Staphylococcus aureus strains, 41 isolates bacteria S. aureus from raw milk (66.1%) and 21 isolates S. aureus from milk filters (33.9%). The presence of antibiotic resistance in Staphylococcus aureus isolates was low, most of them were resistant to amoxicilin. According to the results obtained by the PCR method for the methicillin - resistant S. aureus (MRSA), the mecA gene was present in 6 strains (9.7%), 4 isolates obtained from milk samples (6.5%) and 2 isolates from milk filters (3.2%).  These isolates can be considered as a possible source of resistance genes, which can be spread through the food chain. Nowadays, a globally unfavourable increasing trend of prevalence of methicillin resistant staphylococci strains especially Staphylococcus aureus is being observed worldwide. The improper hygiene and poor farm management practices contributed to the presence of S. aureus in the milk. This may have contributed to the high level of S. aureus isolated. Improving the hygienic conditions of the milking environment and utensils may reduce the prevalence of S. aureus in milk. Objective of this study was monitoring of Staphylococcus aureus prevalence and determine the prevalence rate of antimicrobial resistance of S. aureus isolated from raw milk and milk filters in the Czech Republic.

Necidová L., Šťástková Z., Pospíšilová M., Janštová B., Strejček J., Dušková M., Karpíšková R. The aim of this study was to monitor S. aureus growth and toxin production in soft cheese during the technological processing. In model experiments, raw milk was inoculated separately with five S. aureus strains isolated from milk and milk products. All the strains were producers of staphylococcal enterotoxins (SEs) of types A, B, or C. SEs were detected by the enzyme-linked fluorescence assay (ELFA) performed in the MiniVIDAS device. This study has shown that the amount of SEs varied with the tested strains and stages of the technological process. SEs were detected in soft cheese made from pasteurised milk inoculated with 2.9 × 10 5 CFU/g of S. aureus. The prevention of S. aureus contamination and multiplication during the cheese making process is a prerequisite for the production of safe soft cheese. The most important enterotoxin dose build-up factor can be overcome by strict compliance with the cooling requirements during the manufacture, distribution and storage of the product.

This study concentrates on information concerning the microbiological hazards that can be present in raw milk from animal species other than cows. A total of 54 (23 of ovine and 31 of goat) bulk tank milk samples from 10 farms in the Czech Republic were collected in years 2013 - 2014. The sampling was done at regular time intervals during the whole year, with five to eight samples collected from each of the 10 dairy farms involved in the study. All milk samples were collected into sterile sampling bottles and transported in a cooler sampling case to the laboratory for immediate examination. Farms were randomly selected to cover the whole area of the Czech Republic. The prevalence and characteristic of Escherichia coli, Staphylococcus aureus, Salmonella spp., Campylobacter spp. and Listeria monocytogenes was studied. Raw cow's milk can be contaminated by E. coli intramammarily during clinical or subclinical mastitis and either directly through animal feces or indirectly during milk collection through farm employees or the milking equipment. E. coli was detected in 90.3% of the goat milk and 95.7% of the ovine milk samples. The genes encoding Shiga toxins 1 and 2- (stx1, stx2) were not detected and no STEC was identified. The Eae was the detected in 3 (4.6%) isolates. S. aureus was detected in 9 (29.0%) samples of goat milk and 8 (34.8%) samples of ovine milk. A total 12 (57.1%) enterotoxin positive S. aureus were obtained; 6 (28.6%) were positive for the production of sec encoding enterotoxin SEC; in 4 (19.0%) isolates the gene seh was detected; 2 (9.5%) isolates were proven positive for seg (4.8%) and combination seg and sei (4.8%). The presence of MRSA was not detected in the tested samples in our study. L. monocytogenes was detected in 1 (3.2%) samples of goat milk and 1 (4.3%) samples of ovine milk. The serotype (1/2a, 1/2b) was detected in our study. Campylobacter spp. and Salmonella spp. were not isolated from any of the samples. These results form the basis for determining the microbiological quality standards for goat and ovine milk.