To trace evolution of Panton-Valentine leucocidin-positive clonal complex 398 methicillin-resistant Staphylococcus aureus (MRSA) in the Czech Republic, we tested 103 MRSA isolates from humans. Five (4.9%) were Panton-Valentine leucocidin-positive clonal complex 398, sequence types 1232 and 9181. Spread to the Czech Republic may result from travel to or from other countries.

• MeSH
• bakteriální toxiny * biosyntéza   genetika   MeSH
• dějiny 21. století MeSH
• dospělí MeSH
• exotoxiny * genetika   biosyntéza   MeSH
• leukocidiny * genetika   MeSH
• lidé MeSH
• methicilin rezistentní Staphylococcus aureus * genetika   izolace a purifikace   MeSH
• stafylokokové infekce * mikrobiologie   epidemiologie   MeSH
• Check Tag
• dějiny 21. století MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• historické články MeSH
• Geografické názvy
• Česká republika MeSH

BACKGROUND: A novel Panton-Valentine leukocidin (PVL)-positive meticillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC)5-MRSA-IVc ('Sri Lankan' clone) was recently described from Sri Lanka. Similar isolates caused a recent Irish hospital outbreak. AIM: To investigate the international dissemination and diversity of PVL-positive CC5-MRSA-IVc isolates from hospital and community settings using whole-genome sequencing (WGS). METHODS: Core-genome single nucleotide polymorphism (cgSNP) analysis, core-genome multi-locus sequence typing (cgMLST) and microarray-based detection of antimicrobial-resistance and virulence genes were used to investigate PVL-positive CC5-MRSA-IVc (N = 214 including 46 'Sri Lankan' clone) from hospital and community settings in 12 countries over 17 years. Comparators included 29 PVL-positive and 23 PVL-negative CC5/ST5-MRSA-I/II/IVa/IVc/IVg/V. RESULTS: Maximum-likelihood cgSNP analysis grouped 209/214 (97.7%) CC5-MRSA-IVc into Clade I; average of 110 cgSNPs between isolates. Clade III contained the five remaining CC5-MRSA-IVc; average of 92 cgSNPs between isolates. Clade II contained seven PVL-positive CC5-MRSA-IVa comparators, whereas the remaining 45 comparators formed an outlier group. Minimum-spanning cgMLST analysis revealed a comparably low average of 57 allelic differences between all CC5/ST5-MRSA-IVc. All 214 CC5/ST5-MRSA-IVc were identified as 'Sri Lankan' clone, predominantly spa type t002 (186/214) with low population diversity and harboured a similar range of virulence genes and variable antimicrobial-resistance genes. All 214 Sri Lankan clone isolates and Clade II comparators harboured a 9616-bp chromosomal PVL-encoding phage remnant, suggesting both arose from a PVL-positive meticillin-susceptible ancestor. Over half of Sri Lankan clone isolates were from infections (142/214), and where detailed metadata were available (168/214), most were community associated (85/168). CONCLUSIONS: Stable chromosomal retention of pvl may facilitate Sri-Lankan clone dissemination.

• MeSH
• exotoxiny genetika   MeSH
• leukocidiny genetika   MeSH
• lidé MeSH
• methicilin rezistentní Staphylococcus aureus * MeSH
• methicilin MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• stafylokokové infekce * epidemiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

This study is aimed at detecting and characterizing methicillin-resistant Staphylococcus aureus (MRSA) from bulk tank milk samples of cows, sheep, and goats collected from dairy farms in the Czech Republic. All MRSA isolates were identified using PCR detection of the Staphylococcus aureus-specific fragment SA442 and mecA gene. The staphylococcal chromosomal cassettes mec (SCCmec), spa, and multilocus sequence types (MLST) were determined. The presence of genes encoding enterotoxins (ses), Panton-Valentine leukocidin (pvl), exfoliative toxins A, B (eta, etb), and toxic shock syndrome toxin (tst) were assessed. To differentiate human and animal origin, the presence of staphylokinase (sak) gene, ϕSa3 prophage, and susceptibility to tetracycline was tested. Out of 49 bulk tank milk samples examined, 14 (28.6%) were MRSA-positive. Eleven positive samples came from cow's milk (38%) and the remaining three from goat's milk (33%). All samples of ewe's milk were negative. In MRSA isolates three sequence types containing seven spa types were identified. Twelve isolates (85.7%) belonged to ST398 spa types t011/SCCmec IVa, t011/SCCmec V, t034/SCCmec V, t1456/SCCmec IVa, t1255/SCCmec V, and t2346/SCCmec V. Another two isolates belonged to ST5/t3598/SCCmec IVa and ST8/t064/SCCmec IVNT. In six isolates, one or more ses genes (seb, sed, seg, sei, and sej) were confirmed. One isolate from cow's milk harbored the tst gene. Another two isolates (ST398/t1456/SCCmec IVa and ST5/t3598/SCCmec IVa) harbored the sak gene and ϕSa3 prophage, and the latter was the only tetracycline-susceptible isolate in this study. However, none of the isolates was positive for pvl or eta, etb. These results suggest that there is the wide geographical spread of ST398 across different regions of the Czech Republic with no host preference among dairy cattle and goats. Therefore, when evaluating the occupational and foodborne risks, MRSA carriage and infection should be taken into account.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny genetika   MeSH
• bakteriální toxiny genetika   MeSH
• exotoxiny genetika   MeSH
• farmy MeSH
• kozy MeSH
• leukocidiny genetika   MeSH
• methicilin rezistentní Staphylococcus aureus genetika   izolace a purifikace   MeSH
• methicilin farmakologie   MeSH
• mlékárenství MeSH
• mléko mikrobiologie   MeSH
• multilokusová sekvenční typizace veterinární   MeSH
• ovce MeSH
• rezistence na methicilin * MeSH
• skot MeSH
• stafylokokové infekce epidemiologie   mikrobiologie   veterinární   MeSH
• techniky typizace bakterií veterinární   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVES: Staphylococcus aureus (SA) represents one of the most important microorganism that is part of the normal microflora of humans, but in certain conditions can cause very serious infections. Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for a wide spectrum of nosocomial and community associated infections worldwide. The aim of this study was to determine community acquired MRSA (CA-MRSA), as well as the frequency of Panton-Valentine leukocidin (PVL) genes and staphylococcal cassette chromosome mec (SCCmec) types in isolates obtained from outpatients in the region of 700,000 people (Canton Sarajevo, Bosnia and Herzegovina) Methods: Our investigation included phenotypic and genotypic markers such as antimicrobial resistance, pulsed-field gel electrophoresis (PFGE), SCC typing, and PVL detection. RESULTS: Antimicrobial susceptibility: all MRSA isolates were resistant to the β-lactam antibiotics tested, and all isolates were susceptible to trimethoprim sulphamethoxazole, rifampicin, fusidic acid, linezolid, and vancomycin. After the PFGE analysis, the isolates were grouped into five similarity groups: A-E. The largest number of isolates belonged to one of two groups: C - 60% and D - 27%. In both groups C and D, SCCmec type IV was predominant (60% and 88.8%, respectively). A total of 24% of the isolates had positive expression of PVL genes, while 76% showed a statistically significantly greater negative expression of PVL genes. CONCLUSIONS: Using combination techniques, we were able to investigate the origin and genetic background of the strains. PFGE analysis revealed two large, genetically related groups of strains consisting of 87 isolates. Our results suggest failure to apply the screening policy, and a lack of knowledge about multiresistant MRSA strains. This study showed the local epidemiological situation which should be the basis of antimicrobial empiric therapy for non-hospitalized patients.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální proteiny MeSH
• bakteriální toxiny genetika   MeSH
• chromozomy MeSH
• exotoxiny genetika   MeSH
• infekce získané v komunitě epidemiologie   mikrobiologie   MeSH
• leukocidiny genetika   MeSH
• lidé MeSH
• methicilin rezistentní Staphylococcus aureus účinky léků   genetika   izolace a purifikace   MeSH
• methicilin MeSH
• mikrobiální testy citlivosti MeSH
• proteiny vázající penicilin MeSH
• stafylokokové infekce epidemiologie   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Bosna a Hercegovina MeSH

Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae accounts for serious economic losses in the pig farming industry worldwide. We examined here the immunogenicity and protective efficacy of the recombinant type IV fimbrial subunit protein ApfA as a single antigen vaccine against pleuropneumonia, or as a component of a multi-antigen preparation comprising five other recombinant antigens derived from key virulence factors of A. pleuropneumoniae (ApxIA, ApxIIA, ApxIIIA, ApxIVA and TbpB). Immunization of pigs with recombinant ApfA alone induced high levels of specific serum antibodies and provided partial protection against challenge with the heterologous A. pleuropneumoniae serotype 9 strain. This protection was higher than that engendered by vaccination with rApxIVA or rTbpB alone and similar to that observed after immunization with the tri-antigen combination of rApxIA, rApxIIA and rApxIIIA. In addition, rApfA improved the vaccination potential of the penta-antigen mixture of rApxIA, rApxIIA, rApxIIIA, rApxIVA and rTbpB proteins, where the hexa-antigen vaccine containing rApfA conferred a high level of protection on pigs against the disease. Moreover, when rApfA was used for vaccination alone or in combination with other antigens, such immunization reduced the number of pigs colonized with the challenge strain. These results indicate that ApfA could be a valuable component of an efficient subunit vaccine for the prevention of porcine pleuropneumonia.

• MeSH
• Actinobacillus pleuropneumoniae genetika   imunologie   patogenita   MeSH
• bakteriální adheziny genetika   imunologie   MeSH
• bakteriální fimbrie genetika   imunologie   MeSH
• bakteriální vakcíny genetika   imunologie   MeSH
• Escherichia coli genetika   MeSH
• exotoxiny genetika   metabolismus   MeSH
• faktory virulence genetika   imunologie   MeSH
• infekce bakteriemi rodu Actinobacillus imunologie   prevence a kontrola   veterinární   MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat imunologie   prevence a kontrola   MeSH
• pleuropneumonie imunologie   prevence a kontrola   veterinární   MeSH
• polymerázová řetězová reakce veterinární   MeSH
• prasata MeSH
• rekombinantní proteiny genetika   imunologie   MeSH
• sekvenční analýza DNA veterinární   MeSH
• vakcinace veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH

AIMS: Staphylococcus aureus is an important pathogen characterised by its potential to express many virulence factors. Currently, special attention is being paid to methicillin-resistant strains of S. aureus (MRSA). The aim of this study was to compare the prevalence of 13 selected virulence factor genes in methicillin-resistant versus methicillin-susceptible S. aureus (MSSA) isolates and to investigate their accumulation in the same isolate. METHODS: Real-time PCR was used to detect the presence of genes in 200 isolates of S. aureus (100 MRSA and 100 MSSA) from the University Hospital Olomouc collected in 2005-2006. RESULTS: Six out of the 13 monitored genes were detected more frequently in MRSA isolates: sea, seb, seg, sei, sej and eta, coding for the production of the enterotoxins A, B, G, I, J and the exfoliative toxin A. On the other hand, the pvl and tst genes coding for Panton-Valentine leukocidin and TSST-1 were more frequent in MSSA. Statistical analysis (chi-squared test) of the prevalence of virulence factors in the two groups showed a significant difference (P<0.05) in two cases (seg, sei). CONCLUSIONS: A higher prevalence of selected virulence genes was not confirmed in the methicillin-resistant S. aureus group. This indicates no further increase in their threat.

• MeSH
• bakteriální toxiny genetika   MeSH
• enterotoxiny genetika   MeSH
• exfoliatiny genetika   MeSH
• exotoxiny genetika   MeSH
• financování organizované MeSH
• frekvence genu MeSH
• leukocidiny genetika   MeSH
• methicilin rezistentní Staphylococcus aureus genetika   MeSH
• nemocnice univerzitní MeSH
• Staphylococcus aureus genetika   MeSH
• superantigeny genetika   MeSH
• Geografické názvy
• Česká republika MeSH