Introduction.Staphylococcus aureus (SA) and Staphylococcus epidermidis (SE) are the most common pathogens from the genus Staphylococcus causing biofilm-associated infections. Generally, biofilm-associated infections represent a clinical challenge. Bacteria in biofilms are difficult to eradicate due to their resistance and serve as a reservoir for recurring persistent infections.Gap Statement. A variety of protocols for in vitro drug activity testing against staphylococcal biofilms have been introduced. However, there are often fundamental differences. All these differences in methodical approaches can then be reflected in the form of discrepancies between results.Aim. In this study, we aimed to develop optimal conditions for staphylococcal biofilm formation on pegs. The impact of peg surface modification was also studied.Methodology. The impact of tryptic soy broth alone or supplemented with foetal bovine serum (FBS) or human plasma (HP), together with the impact of the inoculum density of bacterial suspensions and the shaking versus the static mode of cultivation, on total biofilm biomass production in SA and SE reference strains was studied. The surface of pegs was modified with FBS, HP, or poly-l-lysine (PLL). The impact on total biofilm biomass was evaluated using the crystal violet staining method and statistical data analysis.Results. Tryptic soy broth supplemented with HP together with the shaking mode led to crucial potentiation of biofilm formation on pegs in SA strains. The SE strain did not produce biofilm biomass under the same conditions on pegs. Preconditioning of peg surfaces with FBS and HP led to a statistically significant increase in biofilm biomass formation in the SE strain.Conclusion. Optimal cultivation conditions for robust staphylococcal biofilm formation in vitro might differ among different bacterial strains and methodical approaches. The shaking mode and supplementation of cultivation medium with HP was beneficial for biofilm formation on pegs for SA (ATCC 29213) and methicillin-resistant SA (ATCC 43300). Peg conditioning with HP and PLL had no impact on biofilm formation in either of these strains. Peg coating with FBS showed an adverse effect on the biofilm formation of these strains. By contrast, there was a statistically significant increase in biofilm biomass production on pegs coated with FBS and HP for SE (ATCC 35983).

• MeSH
• bakteriologické techniky přístrojové vybavení   metody   MeSH
• biofilmy klasifikace   účinky léků   růst a vývoj   MeSH
• biomasa MeSH
• druhová specificita MeSH
• kultivační média chemie   farmakologie   MeSH
• lidé MeSH
• matrix extracelulárních polymerních látek klasifikace   účinky léků   MeSH
• Staphylococcus klasifikace   účinky léků   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The intestinal microbiota has enormous impact on the health and performance of horses. Staphylococci belong in the phylum Firmicutes, and their occurrence, especially of methicillin-resistant strains and species, has been reported in horses previously. Moreover, biofilm formation is one of the virulence factors; it has been not completely studied in fecal coagulase-negative staphylococci (CoNS) from horses. Therefore, this study was focused on biofilm formation by various species of fecal CoNS from horses because it has been never reported before. In addition, their antibiotic profile was tested. Horses (42) of various breeds from Slovakia/Poland were sampled. Variability in the species of CoNS was detected in feces of horses. Thirty-two strains were identified by using the MALDI-TOF system and classified into nine species and three subspecies of CoNS: Staphylococcus capitis, S. cohnii subsp. cohnii, S. cohnii subsp. urealyticus, S. cohnii subsp. casei, S. epidermidis, S. haemolyticus, S. pasteuri, S. sciuri, S. vitulinus, S. warneri, and S. xylosus. The most frequent species was S. vitulinus. Twenty-two strains showed high biofilm production; 10 strains showed low-grade biofilm production. The highest biofilm formation was measured in the species S. xylosus. Eleven strains (of 32) were methicillin-resistant; the others were susceptible to methicillin.

• MeSH
• antibakteriální látky farmakologie   MeSH
• biofilmy růst a vývoj   MeSH
• druhová specificita MeSH
• feces mikrobiologie   MeSH
• koagulasa nedostatek   MeSH
• koně MeSH
• kyselina mléčná metabolismus   MeSH
• mikrobiální testy citlivosti MeSH
• rezistence na methicilin MeSH
• stafylokokové infekce epidemiologie   mikrobiologie   veterinární   MeSH
• Staphylococcus klasifikace   účinky léků   izolace a purifikace   fyziologie   MeSH
• střevní mikroflóra MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Polsko MeSH
• Slovenská republika MeSH

Staphylococcus pseudintermedius is a species often isolated from animals, as a common element of their microbiota or an agent of infection, and from people associated with an animal habitat, including owners of home pets-dogs and cats. As with many other species, adaptation of these bacteria to the human body can occur, and they become important human pathogens. 59 S. pseudintermedius strains were investigated in this study to determine the factors contributing to human body colonization: inhibition growth of human skin residents isolated from human skin (Staphylococcus epidermidis, Corynebacterium spp., Cutibacterium acnes (formerly Propionibacterium acnes)), biofilm formation, and the presence of ten genes encoding infection-promoting features (including ebpS, spsE, lukS, lukF, pvl, lip, hlgA, hlgB). The ability of human skin to be colonized and the presence of genes that promote the development of skin infections showed the significant potential of the studied strains in their adaptation to the host. However, while a comparison of the characteristics of animal strains and those isolated from human infections does not allow us to claim that we are the witnesses of the speciation of a new human pathogen, it does indicate their gradual adaptation to the human organism.

• MeSH
• bakteriální geny MeSH
• fenotyp MeSH
• kočky MeSH
• lidé MeSH
• mikrobiota MeSH
• psi MeSH
• stafylokokové infekce mikrobiologie   přenos   MeSH
• Staphylococcus klasifikace   fyziologie   MeSH
• virulence MeSH
• zoonózy mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Two Gram-stain-positive, coagulase-negative staphylococcal strains were isolated from abiotic sources comprising stone fragments and sandy soil in James Ross Island, Antarctica. Here, we describe properties of a novel species of the genus Staphylococcus that has a 16S rRNA gene sequence nearly identical to that of Staphylococcus saprophyticus However, compared to S. saprophyticus and the next closest relatives, the new species demonstrates considerable phylogenetic distance at the whole-genome level, with an average nucleotide identity of <85% and inferred DNA-DNA hybridization of <30%. It forms a separate branch in the S. saprophyticus phylogenetic clade as confirmed by multilocus sequence analysis of six housekeeping genes, rpoB, hsp60, tuf, dnaJ, gap, and sod Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and key biochemical characteristics allowed these bacteria to be distinguished from their nearest phylogenetic neighbors. In contrast to S. saprophyticus subsp. saprophyticus, the novel strains are pyrrolidonyl arylamidase and β-glucuronidase positive and β-galactosidase negative, nitrate is reduced, and acid produced aerobically from d-mannose. Whole-genome sequencing of the 2.69-Mb large chromosome revealed the presence of a number of mobile genetic elements, including the 27-kb pseudo-staphylococcus cassette chromosome mec of strain P5085T (ψSCCmecP5085), harboring the mecC gene, two composite phage-inducible chromosomal islands probably essential to adaptation to extreme environments, and one complete and one defective prophage. Both strains are resistant to penicillin G, ampicillin, ceftazidime, methicillin, cefoxitin, and fosfomycin. We hypothesize that antibiotic resistance might represent an evolutionary advantage against beta-lactam producers, which are common in a polar environment. Based on these results, a novel species of the genus Staphylococcus is described and named Staphylococcus edaphicus sp. nov. The type strain is P5085T (= CCM 8730T = DSM 104441T).IMPORTANCE The description of Staphylococcus edaphicus sp. nov. enables the comparison of multidrug-resistant staphylococci from human and veterinary sources evolved in the globalized world to their geographically distant relative from the extreme Antarctic environment. Although this new species was not exposed to the pressure of antibiotic treatment in human or veterinary practice, mobile genetic elements carrying antimicrobial resistance genes were found in the genome. The genomic characteristics presented here elucidate the evolutionary relationships in the Staphylococcus genus with a special focus on antimicrobial resistance, pathogenicity, and survival traits. Genes encoded on mobile genetic elements were arranged in unique combinations but retained conserved locations for the integration of mobile genetic elements. These findings point to enormous plasticity of the staphylococcal pangenome, shaped by horizontal gene transfer. Thus, S. edaphicus can act not only as a reservoir of antibiotic resistance in a natural environment but also as a mediator for the spread and evolution of resistance genes.

• MeSH
• bakteriální geny fyziologie   MeSH
• biologická adaptace genetika   MeSH
• extrémně chladné počasí * MeSH
• extrémní prostředí * MeSH
• genomové ostrovy fyziologie   MeSH
• Staphylococcus klasifikace   genetika   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Antarktida MeSH

Skin microbiome main cultivable aerobes in human are coagulase-negative staphylococci and lipophilic corynebacteria. Staphylococcus strains (155) belonging to 10 species and 105 strains of Corynebacterium belonging to nine species from the skin swabs of healthy male volunteers were investigated to determine their enzymatic activity to main metabolic substrates: carbohydrates, proteins, lipids, and response to factors present on the skin such as osmotic pressure, pH, and organic acids. The results showed that lipophilic corynebacteria have different capacity for adaptation on the skin than staphylococci. Most of Corynebacterium spp. expressed lack of proteinase, phospholipase, and saccharolytic enzymes activity. Corynebacteria were also more sensitive than Staphylococcus spp. to antimicrobial agents existing on human skin, especially to low pH. These characters can explain domination of Staphylococcus genera on healthy human skin. It can be suggested that within these two bacterial genus, there exists conceivable cooperation and reciprocal protection which results in their quantitative ratio. Such behavior must be considered as crucial for the stability of the population on healthy skin.

• MeSH
• bakteriální proteiny metabolismus   MeSH
• Corynebacterium klasifikace   enzymologie   izolace a purifikace   fyziologie   MeSH
• dospělí MeSH
• kůže mikrobiologie   MeSH
• lidé MeSH
• Staphylococcus klasifikace   enzymologie   izolace a purifikace   fyziologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

A cover-glass placed on a heavily inoculated culture plate clearly differentiates facultatively anaerobic staphylococci growing underneath the cover-glass after overnight incubation from nongrowing aerobic micrococci. Even if there are some exceptions, all medically significant staphylococci can grow in the test. Thus, the test provides a cost-effective and highly specific tool for separation of both genera which fundamentally differ in their pathogenicity.

• MeSH
• aerobióza MeSH
• anaerobióza MeSH
• bakteriologické techniky ekonomika   metody   MeSH
• laboratoře MeSH
• lidé MeSH
• Micrococcus klasifikace   izolace a purifikace   fyziologie   MeSH
• náklady a analýza nákladů MeSH
• senzitivita a specificita MeSH
• sklo MeSH
• Staphylococcus klasifikace   izolace a purifikace   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

V období 2005–2009 bylo vyšetřeno celkem 87 kmenů S. saprophyticus pocházejících z močí pacientů s diagnózou zánět močového měchýře – cystitis. U všech těchto kmenů byla sledována rezistence k vankomycinu, nitrofurantoinu, doxycyklinu, oxacilinu, amoxicilin/klavulanátu, cefoxitinu a sulfamethoxazol/trimethoprimu. Tvorba biofilmu byla detekována pomocí modifikované Christensenovy metody. Nebyl nalezen žádný kmen rezistentní k vankomycinu, k nitrofurantoinu byly rezistentní 2 kmeny (2,3 %), k doxycyklinu 9 kmenů (10,3 %), k oxacilinu 20 kmenů (23,0 %), k amoxicilinu s klavulanátem a k cefoxitinu shodně 6 kmenů (6,9 %), ke sulfamethoxazol/trimethoprimu 1 kmen (1,1 %). Z 20 375 kultivačně pozitivních močí testovaných v letech 2005-2009 v naší laboratoři byl S. saprophyticus prokázán jako původce 0,4 % zánětů močového měchýře. Většina kmenů (67) pocházela od žen (77,0 %), zejména mladšího věku. Biofilm tvořilo 16 z 87 kmenů (18,4 %).

Eighty-seven Staphylococcus saprophyticus strains isolated from urine of 87 patients with cystitis were examined in 2005–2009. All strains were tested for resistance to vancomycin, nitrofurantoin, doxycycline, oxacillin, amoxicillin/clavulanate, cefoxitin and sulfamethoxazole/trimethoprim and for biofilm formation by a modified Christensen method. None of the tested strains of S. saprophyticus showed resistance to vancomycin, while 2 strains (2.3 %) were resistant to nitrofurantoin, 9 (10.3%) to doxycycline, 20 (23.0 %) to oxacillin, 6 (6.9%) to amoxicillin/clavulanate, 6 (6.9%) to cefoxitin and 1 (1.1%) to sulfamethoxazole/trimethoprim. S. saprophyticus was detected as the causative agent of cystitis in 0.4 % of 20,375 culture positive urine samples analyzed in our laboratory between 2005 and 2009. Most 67 (77.0%) S. saprophyticus strains were recovered from women, particularly from young women. Biofilm formation was detected in 16 (18.4 %) out of 87 S. saprophyticus strains.

• Klíčová slova
• S. saprophyticus, močové infekce, biofilm, antibiotika,
• MeSH
• bakteriální léková rezistence MeSH
• biofilmy růst a vývoj   MeSH
• cystitida mikrobiologie   MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• moč mikrobiologie   MeSH
• Staphylococcus fyziologie   účinky léků   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

The ability of surfactants obtained from three Lactobacillus acidophilus strains to inhibit Staphylococcus aureus and S. epidermidis biofilms was evaluated. Their influence was determined on bacterial initial adhesion, biofilm formation and dispersal using MTT-reduction assay, confocal laser scanning microscopy and image PHLIP analysis. The number of adhering S. aureus and S. epidermidis cells after a 3-h co-incubation with biosurfactants was reduced by 5-56 % in a strain-and dose-dependent manner. S. epidermidis-and, to a lower extent, in S. aureus-biofilm formation was also inhibited in the presence of the tested surfactants. The addition of surfactants to preformed mature biofilms accelerated their dispersal, and changed the parameters of biofilm morphology. The L. acidophilus-derived surfactants inhibit bacterial deposition rate and biofilm development (and also its maturation) without

• MeSH
• bakteriální adheze účinky léků   MeSH
• biofilmy růst a vývoj   účinky léků   MeSH
• Lactobacillus acidophilus metabolismus   MeSH
• lidé MeSH
• povrchově aktivní látky farmakologie   metabolismus   MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus fyziologie   izolace a purifikace   účinky léků   MeSH
• Check Tag
• lidé MeSH

Five isolates of coagulase-negative staphylococci were obtained from human urine, the gastrointestinal tract of squirrel monkeys, pig skin and from the environment. All key biochemical characteristics of the tested strains corresponded with the description of Staphylococcus xylosus species. However, partial 16S rRNA gene sequences obtained from analysed strains corresponded with those of Staphylococcus nepalensis reference strains, except for two strains which differed in one residue. Ribotyping with EcoRI and HindIII restriction enzymes, whole cell protein profile analysis performed by SDS-PAGE and SmaI macrorestriction analysis were used for more precise characterization and identification of the analysed strains. Obtained results showed that EcoRI and HindIII ribotyping and whole cell protein fingerprinting are suitable and reliable methods for the differentiation of S. nepalensis strains from the other novobiocin resistant staphylococci, whereas macrorestriction analysis was found to be a good tool for strain typing. The isolation of S. nepalensis is sporadic, and according to our best knowledge this study is the first report of the occurrence of this species in human clinical material as well as in other sources.

• MeSH
• DNA bakterií genetika   chemie   MeSH
• DNA fingerprinting MeSH
• financování organizované MeSH
• fylogeneze MeSH
• lidé MeSH
• novobiocin MeSH
• ribotypizace MeSH
• RNA ribozomální 16S analýza   genetika   MeSH
• sekvenční analýza DNA MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus fyziologie   genetika   izolace a purifikace   klasifikace   MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH

A polyphasic identification approach was applied to a group of 11 novobiocin-resistant staphylococci isolated from human clinical materials. Phenotypic characteristics obtained by both commercial and conventional tests assigned eight strains as Staphylococcus xylosus and three strains as ambiguous S. xylosus/Staphylococcus equorum. In contrast to biotyping, ribotyping with EcoRI and HindIII restriction endonucleases and whole-cell protein fingerprinting assigned six analysed strains as S. equorum, and five strains as Staphylococcus succinus. Confirmation of the identification was done by partial 16S rRNA gene sequencing and S. equorum isolates were verified by a PCR assay targeting the sodA gene. From the data it has been implied that ribotyping and whole-cell protein analysis can be used to differentiate between the biochemically almost indistinguishable species S. xylosus, S. equorum and S. succinus. The present study confirms what is believed to be the first occurrence of S. equorum in a relevant human clinical material in the Czech Republic and describes what is believed to be the first-ever isolation of S. succinus from human clinical material.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence MeSH
• bakteriální proteiny analýza   genetika   MeSH
• dítě MeSH
• DNA bakterií genetika   chemie   MeSH
• financování organizované MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• molekulární sekvence - údaje MeSH
• novobiocin farmakologie   MeSH
• polymerázová řetězová reakce MeSH
• proteom analýza   MeSH
• ribotypizace MeSH
• ribozomální DNA genetika   chemie   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• shluková analýza MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus fyziologie   genetika   izolace a purifikace   klasifikace   MeSH
• superoxiddismutasa genetika   MeSH
• techniky typizace bakterií MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH