Some strains of the genus Enterococcus are effective probiotic bacteria if they meet safety and probiotic criteria. In our study, 17 canine enterococci previously selected from a group of 160 isolates based on safety criteria were screened for some functional properties relevant to their use as probiotics. The results of antimicrobial resistance testing showed sensitivity of eleven strains to EFSA recommended antimicrobials. In contrast, the most frequent resistance was observed for cefotaxim (15/17) and oxacillin (13/17). PCR detection of resistance genes (vanA, vanB, vanC, tetM, tetL, ermB, and mefA) revealed the presence of mefA gene in five Enterococcus faecium strains and vanA gene in one strain. The production of enzymes commonly associated with intestinal diseases was in general rare (β-glucosidase 2/17, α-chymotrypsin 1/17, N-acetyl-β-glucosaminidase 0/17, and β-glucuronidase 0/17). The measurement of strain survival rate (%) under the conditions simulating gastric (pH 2.5) and bile juices (0.3% bile) showed considerable differences between strains (< 0.01 to 4.7% after 90 min for gastric juices, 48.0 to 254.0% after 180 min for bile). The concentration of produced L-lactic acid ranged between 83.1 to 119.3 mmol/L after 48 h cultivation depending on the strain. All strains fermented 16 out of 49 different carbohydrates (range from 17 to 23/49). Antimicrobial activity was recorded for two strains against some species of Listeria sp. and Enterococcus sp. Finally, two E. faecium candidates (IK25 and D7) were selected for testing in dogs, and hereafter they could possibly extend the currently limited range of beneficial bacteria of canine origin used as a dietary supplement for dogs.
- MeSH
- antibakteriální látky farmakologie normy MeSH
- Bacteria účinky léků MeSH
- bakteriální geny MeSH
- bakteriální léková rezistence genetika MeSH
- bakteriociny genetika MeSH
- Enterococcus účinky léků genetika metabolismus fyziologie MeSH
- kyselina mléčná biosyntéza MeSH
- metabolismus sacharidů MeSH
- mikrobiální testy citlivosti MeSH
- probiotika farmakologie MeSH
- psi MeSH
- žaludeční kyselina MeSH
- žluč MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Enterococci form a complex, diverse, and very important group of bacteria from the technological and food safety aspect, or from the health-improving aspect as probiotics. Generally, enterococci are considered to be of low pathogenic potential, which is associated mostly with clinical strains. In these strains, production of virulence factors as well as resistance to many antimicrobial drugs could complicate treatment of nosocomial infections. Because there is a lack of information on incidence of these attributes in animal commensal enterococci, we screened 160 strains originating from feces of clinically healthy dogs in Eastern Slovakia (n = 105). The predominant species were Enterococcus faecium (57.5%) followed by Enterococcus faecalis (21.9%), and Enterococcus hirae (17.5%), while Enterococcus casseliflavus (1.9%) and Enterococcus mundtii (1.2%) rarely occurred. Among the tested antibiotics, gentamicin (high level) was the most effective drug against canine enterococci (95% of isolates were sensitive). In contrast, the highest resistance recorded (71.9%) was to teicoplanin. PCR screening showed the highest incidence of virulence genes in E. faecalis species. The most frequently detected were genes encoding adhesins efa Afm and efa Afs and sex pheromone cpd. IS16 gene, a marker specific for hospital strains, appeared in nine E. faecium strains. No strain was positive for DNase activity, 8.8% of the isolated strains showed gelatinase activity, and almost 100% strains produced tyramine. It seems commensal-derived enterococci from dogs could also to some extent be potential reservoir of risk factors for other microbiota or organisms.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- bakteriální proteiny genetika metabolismus MeSH
- Enterococcus účinky léků genetika izolace a purifikace fyziologie MeSH
- faktory virulence genetika metabolismus MeSH
- mikrobiální testy citlivosti MeSH
- psi mikrobiologie MeSH
- symbióza MeSH
- zvířata MeSH
- Check Tag
- psi mikrobiologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Klíčová slova
- avoparcin,
- MeSH
- bakteriální léková rezistence genetika účinky léků MeSH
- Enterococcus fyziologie genetika klasifikace ultrastruktura MeSH
- enterokoky rezistentní vůči vankomycinu * genetika účinky léků ultrastruktura MeSH
- exprese genu genetika účinky léků MeSH
- glykopeptidy účinky léků MeSH
- infekce spojené se zdravotní péčí mortalita přenos MeSH
- lidé MeSH
- vankomycin MeSH
- zoonózy přenos MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Staphylococcus aureus is a major food-borne pathogen due to the production of enterotoxin and is particularly prevalent in contaminated milk and dairy products. The lactic acid bacteria (LAB) are widely used as biocontrol agents in fermented foods which can inhibit pathogenic flora. In our work, we investigated the influence of three strains of LAB (Lactobacillus plantarum, Streptococcus thermophilus and Enterococcus durans) on the relative expression of three enterotoxin genes (sea, sec, sell) and eight virulence and/or regulatory genes (sarA, saeS, codY, srrA, rot, hld/RNAIII, agrA/RNAII, sigB) in two S. aureus strains (MW2 and Sa1612) in TSB and reduced-fat milk (1.5 %) at 30 °C over a 24-h period. The tested LAB and S. aureus strains proved to be mutually non-competitive or only slightly competitive during co-cultivation. In addition, under the above-mentioned conditions, differential gene expression between the S. aureus MW2 and Sa1612 strains was well documented. S. aureus growth was changed in mixed culture with LAB; however, its effect on the repression of sea and sec expression correlated with production of these virulence factors. In comparison, the presence of LAB strains generally inhibited the expression of sec, sell, sarA, seaS, agrA/RNAII and hld/RNAIII genes. The effect of LAB strains presence on the expression of sea, codY, srrA, rot and sigB genes was medium, time, LAB and S. aureus strain specific. SEA and SEC production was significantly reduced in milk compared to TSB in pure culture. After the 24-h cultivation, S. aureus MW2 and Sa1612 SEC production was 187 and 331 times lower in milk compared to TSB, respectively (0.07 and 0.39 ng/mL in milk, versus 13.1 and 129.2 ng/mL in TSB, respectively). At the same time S. aureus MW2 and Sa1612 SEA production was 77 and 68 times lower in milk compared to TSB, respectively (0.99 and 0.17 ng/mL in milk, versus 76.4 and 11.5 ng/mL in TSB, respectively). This study has revealed new insights into the interaction between S. aureus and LAB (L. plantarum, S. thermophilus, E. durans) on the level of the expression and/or production of S. aureus enterotoxins, regulatory and virulence genes in different media, including milk. This study provides data which may improve the quality of food production.
- MeSH
- Enterococcus fyziologie MeSH
- enterotoxiny biosyntéza genetika MeSH
- faktory virulence genetika MeSH
- kokultivační techniky MeSH
- kultivační média MeSH
- Lactobacillus plantarum fyziologie MeSH
- mikrobiální interakce MeSH
- mléko * MeSH
- potravinářská mikrobiologie MeSH
- regulační geny MeSH
- Staphylococcus aureus genetika růst a vývoj metabolismus patogenita MeSH
- Streptococcus thermophilus fyziologie MeSH
- transkriptom MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
It is demonstrated that numerous bacteria are able to attach to surfaces of equipment used for food handling or processing. In this study, a strain of Enterococcus durans, originally isolated from a milking machine surface, was firstly studied for its biofilm formation potential on plastic and stainless steel supports. The strain was found to be a biofilm producer either at 25, 30 or 37 °C on polystyrene microtitre plates, with a best adherence level observed at 25 °C. En. durans showed a strong adhesion to stainless steel AISI-304. Antibacterial and anti-adherence activities of En. durans were tested against four foodborne pathogens (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853 and Listeria innocua CLIP 74915) which were shown as biofilm producers on both plastic and stainless steel. En. durans cells and cell-free culture supernatant showed a significant (P < 0.05) inhibition potential of the pathogens either on solid media or in broth co-cultures. Characterization of the antibacterial substances indicated their proteinaceous nature which assigned them most probably to bacteriocins group.
- MeSH
- bakteriální adheze účinky léků MeSH
- bakteriální proteiny metabolismus MeSH
- biofilmy účinky léků MeSH
- biologické přípravky metabolismus MeSH
- Enterococcus metabolismus fyziologie MeSH
- gramnegativní bakterie účinky léků fyziologie MeSH
- grampozitivní bakterie účinky léků fyziologie MeSH
- mikrobiologie životního prostředí * MeSH
- nerezavějící ocel MeSH
- plastické hmoty MeSH
- potravinářská mikrobiologie MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
This study's aims were to assess the prevalence of, and to characterize, vancomycin-resistant enterococci (VRE) from rooks (Corvus frugilegus) wintering in Europe during 2010/2011. Faeces samples were cultivated selectively for VRE and characterized. Pulsed-field gel electrophoresis and multilocus sequence typing (MLST) were used to examine epidemiologic relationships of vanA-containing VRE. The vanA-carrying VRE were tested in vitro for mobility of vancomycin resistance traits. VRE were found in 62 (6%) of 1073 rook samples. Enterococcal species diversity comprised Enterococcus gallinarum (48 isolates), followed by E. faecium (9) and E. faecalis (5). Eight VRE harboured the vanA and ermB genes. Seven vanA-carrying VRE originated from the Czech Republic and one from Germany. All vanA-carrying VRE were identified as E. faecium. Based on MLST analysis, six vanA-positive isolates were grouped as ST92 type, one isolate belonged to ST121, and the remaining one was described as a novel type ST671. Seven out of eight isolates were able to transfer the vancomycin resistance trait via filter mating with a transfer rate of 8.95 ± 3.25 × 10(-7) transconjugants per donor. In conclusion, wintering rooks in some European countries may disseminate clinically important enterococci and pose a risk for environmental contamination.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální proteiny genetika MeSH
- Enterococcus účinky léků genetika izolace a purifikace fyziologie MeSH
- feces mikrobiologie MeSH
- ligasy tvořící vazby C-O genetika MeSH
- mikrobiální testy citlivosti MeSH
- multilokusová sekvenční typizace MeSH
- přenos genů horizontální genetika MeSH
- pulzní gelová elektroforéza MeSH
- rezistence na vankomycin * genetika MeSH
- shluková analýza MeSH
- vankomycin farmakologie MeSH
- vrány genetika mikrobiologie MeSH
- zdroje nemoci mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
Eight Gram-positive, catalase-negative bacterial strains were isolated during screening of enterococcal populations on plants. rep-PCR fingerprinting using the (GTG)(5) primer showed that the isolates constituted a single cluster that was separate from all known enterococcal species. 16S rRNA gene sequence phylogenetic analysis of three representative strains showed that the isolates belonged to the genus Enterococcus and that they clustered with the Enterococcus faecalis species group. Sequencing of the genes for the phenylalanyl-tRNA synthase alpha subunit (pheS) and the RNA polymerase alpha subunit (rpoA) also revealed the isolates' separate taxonomic position. Application of whole-cell protein fingerprinting, automated ribotyping and extensive phenotyping demonstrated the genetic and phenotypic homogeneity of the isolates and confirmed their separate position within the E. faecalis species group. The isolates represent a novel species of the genus Enterococcus, for which the name Enterococcus plantarum sp. nov. is proposed; the type strain is CCM 7889(T) (=LMG 26214(T)=C27(T)).
- MeSH
- DNA bakterií chemie genetika MeSH
- DNA řízené RNA-polymerasy genetika MeSH
- Enterococcus klasifikace genetika izolace a purifikace fyziologie MeSH
- fenylalanin-tRNA-ligasa genetika MeSH
- fylogeneze MeSH
- katalasa metabolismus MeSH
- molekulární sekvence - údaje MeSH
- molekulární typizace MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- rostliny mikrobiologie MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- techniky typizace bakterií MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Three enterococci constituted two aberrant branches after numerical analysis of (GTG)5-PCR fingerprints: analogous patterns were found for two water isolates, strains W213 and W442T, and a separate position was found for an isolate from the gut of a termite, strain LMG 8895T. 16S rRNA gene sequence analysis classified all three strains in the Enterococcus faecalis species group. Further sequencing analysis of the housekeeping gene pheS (encoding the phenylalanyl-tRNA synthase alpha-subunit) and whole-cell-protein analysis confirmed a distinct position for the two water isolates and the termite strain, respectively. DNA-DNA hybridization experiments and distinct phenotypic features between the strains studied and representatives of the E. faecalis species group confirmed novel species status, respectively, for the two water isolates, strains W213 and W442T, and for strain LMG 8895T. The names Enterococcus silesiacus sp. nov. and Enterococcus termitis sp. nov. are proposed for the novel taxa, with W442T (= CCM 7319T = LMG 23085T) and LMG 8895T (= CCM 7300T) as the respective type strains.
- MeSH
- antibakteriální látky farmakologie MeSH
- Enterococcus fyziologie izolace a purifikace MeSH
- extracelulární matrix mikrobiologie MeSH
- fibrinogen metabolismus MeSH
- fibronektiny metabolismus MeSH
- laktoferrin metabolismus MeSH
- lidé MeSH
- transferin metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
