Cíl práce: Detekovat a charakterizovat kmeny meticilin-rezistentních bakterií Staphylococcus aureus (MRSA) v chovech prasat, na porážce a ve vzorcích masa v tržní síti. Materiál a metody: Celkem bylo v letech 2013-2015 vyšetřeno 890 vzorků, z prvovýroby (prasat na farmách) pocházelo 59 vzorků, ze zpracovatelských podniků (jatek) pocházelo 463 stěrů a z tržní sítě bylo odebráno 368 vzorků vepřového masa a jater. K detekci MRSA byla použita kultivační metoda. Po homogenizaci vzorků bylo provedeno neselektivní pomnožení v pufrované peptonové vodě při 37 °C po dobu 18-24 hodin a dále bylo provedeno dvoustupňové selektivní pomnožení. Pět mililitrů PPV bylo přeneseno do média Mueller-Hinton s 6,5% přídavkem NaCl a po inkubaci, která probíhala ve všech krocích při 37 °C po dobu 18-24 hodin, byl 1 ml inokulován do trypton-sojového bujónu s cefoxitinem a aztreonamem. Další den pak byla suspenze vyočkována na Baird-Parker a Brilliance TM MRSA 2 agary. Suspektní kolonie byly konfirmovány metodou PCR, byl detekován fragment SA442 specifický pro kmeny druhu S. aureus, mecA gen kódující rezistenci k meticilinu a bylo provedeno potvrzení příslušnosti izolátu ke klonálnímu komplexu CC398. Dále byla stanovena rezistence k panelu 11 antimikrobiálních látek za použití diskové difuzní metody. Výsledky: V rámci studie bylo získáno 51 kmenů MRSA, 15 pocházelo od živých prasat, 31 izolátů z jatek a 5 bylo detekováno ve vzorcích z tržní sítě. Ke klonálnímu komplexu CC398 náleželo 47 (92,2 %) kmenů MRSA. Čtyři izoláty non-CC398 byly získány z prostředí dvou porážek a pocházely od zvířat ze tří různých farem. Izoláty často vykazovaly vícečetnou rezistenci. U izolátů MRSA byla detekována rezistence k erytromycinu (36; 70,6 %), tetracyklinu (29; 56,9 %), fluorochinolonům (7; 13,7 %), ko-trimoxazolu (6; 11,8 %) a aminoglykosidům (4; 7,8 %). Závěr. V potravinách živočišného původu byly detekovány izoláty MRSA náležející do klonálního komplexu CC398, jenž pochází z chovů hospodářských zvířat. Tyto kmeny jsou často charakterizovány vícečetnou rezistencí. Role potravního řetězce v šíření LA-MRSA zatím není jednoznačně objasněna.

Objectives: To detect and characterize strains of methicillin-resistant Staphylococcus aureus (MRSA) on pig farms, at slaughterhouses and meat samples at retail sale. Material and Methods; A total of 890 samples were examined in the years 2013-2015, comprising 59 samples from primary production (pig farms), 463 swabs from processing plants (slaughterhouses) and 368 samples of pork meat and liver collected at retail sale. The culture method was used for the detection of MRSA. After homogenization, the samples were enriched in buffered peptone water at 37 °C for 18-24 hours and two-stage selective enrichment was performed. Five milliliters of PPV were transferred to Mueller-Hinton medium with addition of 6.5% NaCl and after incubation at 37 °C for 18-24 hours, 1 ml was inoculated into tryptone soy broth with cefoxitin and aztreonam. The suspension was plated onto Baird-Parker and Brilliance TM MRSA 2 agar on the next day. Suspected colonies were confirmed by PCR, the specific S. aureus fragment SA442, the mecA gene encoding resistance to methicillin and relation to the clonal complex CC398 were detected. Further, resistance test to a panel of 11 antimicrobial agents using the disk diffusion method was performed. Results: Within this study, 51 MRSA strains were obtained, of which 15 originated from live pigs, 31 isolates were from slaughterhouses and 5 were detected in retail samples. Forty-seven (92.2 %) MRSA strains belonged to the clonal complex CC398. Four non-CC398 isolates were obtained from two slaughterhouses and came from three farms. The strains often showed multiple resistance. In some MRSA isolates, resistance to erythromycin (36; 70.6 %), tetracychne (29; 56.9 %), fluoroquinolones (7; 13.7 %), co-trimoxazole (6; 11.8 %) and aminoglycosides (4; 7.8 %) was detected. Conclusion: MRSA isolates of the clonal complex CC398 dominate in foods of animal origin. These strains originate from livestock and are often characterized by multiple resistance to antimicrobials. The role of the food chain in the spread of LA-MRSA is not yet clearly understood.

• MeSH
• bakteriologické techniky MeSH
• lidé MeSH
• maso mikrobiologie   zásobování a distribuce   MeSH
• methicilin rezistentní Staphylococcus aureus izolace a purifikace   klasifikace   MeSH
• metody pro přípravu analytických vzorků MeSH
• potravní řetězec * MeSH
• prasata * mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Rats are a reservoir of human- and livestock-associated methicillin-resistant Staphylococcus aureus (MRSA). However, the composition of the natural S. aureus population in wild and laboratory rats is largely unknown. Here, 144 nasal S. aureus isolates from free-living wild rats, captive wild rats and laboratory rats were genotyped and profiled for antibiotic resistances and human-specific virulence genes. The nasal S. aureus carriage rate was higher among wild rats (23.4%) than laboratory rats (12.3%). Free-living wild rats were primarily colonized with isolates of clonal complex (CC) 49 and CC130 and maintained these strains even in husbandry. Moreover, upon livestock contact, CC398 isolates were acquired. In contrast, laboratory rats were colonized with many different S.aureus lineages-many of which are commonly found in humans. Five captive wild rats were colonized with CC398-MRSA. Moreover, a single CC30-MRSA and two CC130-MRSA were detected in free-living or captive wild rats. Rat-derived S. aureus isolates rarely harbored the phage-carried immune evasion gene cluster or superantigen genes, suggesting long-term adaptation to their host. Taken together, our study revealed a natural S. aureus population in wild rats, as well as a colonization pressure on wild and laboratory rats by exposure to livestock- and human-associated S.aureus, respectively.

• MeSH
• antibakteriální látky farmakologie   MeSH
• divoká zvířata mikrobiologie   MeSH
• ekosystém MeSH
• faktory virulence genetika   MeSH
• hemokoagulace MeSH
• methicilin farmakologie   MeSH
• molekulární epidemiologie MeSH
• nos mikrobiologie   MeSH
• potkani Sprague-Dawley MeSH
• stafylokokové infekce epidemiologie   veterinární   MeSH
• Staphylococcus aureus účinky léků   genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Německo MeSH

Staphylococcus aureus is an important pathogen of humans and animals. The aim of this study was to characterize 71 of S. aureus isolates from raw cow milk in southern Xinjiang of China, including molecular typing, distribution of virulence genes, and antimicrobial susceptibility. The isolates belonged to 18 sequence types (STs) (including 11 novel STs) and 6 spa types which were divided into five different clonal complexes (CCs), including CC188, CC352, CC22, CC398, and CC5406. The majority of the strains was grouped into multilocus sequence typing (MLST) CC188 (n = 41), t189 (n = 40), and ST5796 (n = 17). Only 30.9, 12.7, 11.3, and 9.9% of the isolates were resistant to erythromycin, clindamycin/norfloxacin, tetracycline, and gentamicin, respectively. Nine of multidrug resistant (MDR) isolates were observed which was associated with CC398-t2876. The adhesion molecules clfa, clfb, and hlb were most frequently detected with the percentage rate of 98.6% (70/71), 98.6% (70/71), and 90.1% (64/71), respectively. The percentage rates of the staphylococcal enterotoxin genes sea, seb, sec, sed, seg, and sei in S. aureus isolates were 5.6, 19.8, 40.8, 1.4, 49.3, and 30.9%, respectively. The see, seh, and sej genes were not found. This study provides data about the occurrence of S. aureus in raw cow milk, revealing high carriage frequency, drug resistance, and population structure of S. aureus. Furthermore, this study suggests that effective hygienic measures be taken when handling dairy cows, in order to prevent spreading MDR strains to human through direct contact and/or consumption of contaminated food.

• MeSH
• antibakteriální látky farmakologie   MeSH
• antibiotická rezistence MeSH
• enterotoxiny genetika   MeSH
• methicilin rezistentní Staphylococcus aureus * MeSH
• mléko MeSH
• multilokusová sekvenční typizace MeSH
• skot MeSH
• stafylokokové infekce * epidemiologie   veterinární   MeSH
• Staphylococcus aureus MeSH
• virulence genetika   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Čína MeSH

Staphylococcus aureus is one of the major causes of bloodstream infections. The aim of our study was to characterize methicillin-resistant Staphylococcus aureus (MRSA) isolates from blood of patients hospitalized in the Czech Republic between 2016 and 2018. All MRSA strains were tested for antibiotic susceptibility, analyzed by spa typing and clustered using a Based Upon Repeat Pattern (BURP) algorithm. The representative isolates of the four most common spa types and representative isolates of all spa clonal complexes were further typed by multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. The majority of MRSA strains were resistant to ciprofloxacin (94%), erythromycin (95.5%) and clindamycin (95.6%). Among the 618 strains analyzed, 52 different spa types were detected. BURP analysis divided them into six different clusters. The most common spa types were t003, t586, t014 and t002, all belonging to the CC5 (clonal complex). CC5 was the most abundant MLST CC of our study, comprising of 91.7% (n = 565) of spa-typeable isolates. Other CCs present in our study were CC398, CC22, CC8, CC45 and CC97. To our knowledge, this is the biggest nationwide study aimed at typing MRSA blood isolates from the Czech Republic.

• Publikační typ
• časopisecké články MeSH

OBJECTIVES: To gain data on the current molecular epidemiology and resistance of MRSA in the Czech Republic. METHODS: Between September 2017 and January 2018, a total of 441 single-patient MRSA isolates were collected from 11 Czech hospitals and analysed by spa typing, SCCmec typing, antibiotic susceptibility testing, detection of the PVL toxin and the arcA gene. RESULTS: Of all MRSA isolates, 81.41% (n = 359) belonged to the CC5-MRSA clone represented by the spa types t003 (n = 136), t586 (n = 92), t014 (n = 81), t002 (n = 20) and other spa types (n = 30); a majority of the CC5 isolates (n = 348, 96.94%) carried SCCmec type II. The occurrence of CC5-MRSA was more likely in older inpatients and associated with a healthcare origin (P < 0.001). The CC5-MRSA isolates were resistant to more antimicrobial drugs compared with the other MRSAs (P < 0.001). Interestingly, t586 was detected in blood samples more often than the other spa types and, contrary to other spa types belonging to CC5-MRSA, t586 was not associated with patients of advanced age. Other frequently found lineages were CC8 (n = 17), CC398 (n = 11) and CC59 (n = 10). The presence of the PVL was detected in 8.62% (n = 38) of the MRSA isolates. CONCLUSIONS: The healthcare-associated CC5-MRSA-II lineage (t003, t586, t014) was found to be predominant in the Czech Republic. t586 is a newly emerging spa type in the Czech Republic, yet reported rarely in other countries. Our observations stress the need for MRSA surveillance in the Czech Republic in order to monitor changes in MRSA epidemiology.

• MeSH
• antibakteriální látky farmakologie   MeSH
• genotyp MeSH
• lidé MeSH
• methicilin rezistentní Staphylococcus aureus * genetika   MeSH
• mikrobiální testy citlivosti MeSH
• molekulární epidemiologie MeSH
• senioři MeSH
• stafylokokové infekce * epidemiologie   MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH