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
- Anti-Bacterial Agents pharmacology standards MeSH
- Bacteria drug effects MeSH
- Genes, Bacterial MeSH
- Drug Resistance, Bacterial genetics MeSH
- Bacteriocins genetics MeSH
- Enterococcus drug effects genetics metabolism physiology MeSH
- Lactic Acid biosynthesis MeSH
- Carbohydrate Metabolism MeSH
- Microbial Sensitivity Tests MeSH
- Probiotics pharmacology MeSH
- Dogs MeSH
- Gastric Acid MeSH
- Bile MeSH
- Animals MeSH
- Check Tag
- Dogs MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Seventy-eight isolates of lactic acid bacteria from Ukraine and Thailand were screened for bacteriocinogenic activity against indicator strain Lactobacillus sakei subsp. sakei JCM 1157. One isolate showed an antagonistic activity of cell-free supernatant eliminated after the treatment with Proteinase K. Based on 16S rRNA gene sequence, this isolate was identified as Enterococcus italicus. Bacteriocin produced by this strain showed antimicrobial activity against L. sakei subsp. sakei JCM 1157, Brochothrix thermosphacta DSMZ 20171, and Listeria ivanovii subsp. ivanovii DSMZ 20750 in agar well diffusion assay. This bacteriocin was cationic and hydrophobic. The partially purified bacteriocin was thermostable, while heating of cell-free supernatant increased its activity more than twofold. Molecular mass of the partially purified bacteriocin as determined by SDS-PAGE differed from enterocin A and B previously known for E. italicus. Concentrated bacteriocin decreased the level of biofilm formation in L. sakei subsp. sakei JCM 1157 and Pseudomonas aeruginosa PAO1 in 52.5 and 48.0%, respectively (p < 0.05). We suggest that the studied bacteriocin could be a perspective antibiofilm agent in food conservation and medicine.
- MeSH
- Anti-Bacterial Agents chemistry metabolism pharmacology MeSH
- Bacteriocins chemistry metabolism pharmacology MeSH
- Biofilms drug effects MeSH
- Brassica microbiology MeSH
- Enterococcus chemistry genetics isolation & purification metabolism MeSH
- Fermentation MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Microbial Sensitivity Tests MeSH
- Pseudomonas aeruginosa drug effects MeSH
- Drug Stability MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Thailand MeSH
OBJECTIVES: Urban wastewater contains various micropollutants and a high number of different micro-organisms. Some bacteria in wastewater can attach to surfaces and form biofilm, which gives bacteria an advantage in the fight against environmental stresses. This work focused on analysis of bacterial communities in biofilms isolated from influent and effluent sewerage of a wastewater treatment plant (WWTP) in Bratislava, Slovakia. METHODS: Detection of biofilm microbiota was performed by culture-independent and -dependent approaches. The composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with construction of 16S rRNA clone libraries. Analysis of the concentration of antibiotics and the prevalence of antibiotic-resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was also studied. RESULTS: Biofilm collected at the inlet point was characterised primarily by the presence of Pseudomonas spp., Acinetobacter spp. and Janthinobacterium spp. clones, whilst members of the genus Pseudomonas were largely detected in biofilm isolated in outflow of the WWTP. Predominant antibiotics such as azithromycin, clarithromycin and ciprofloxacin were found in influent wastewater. The removal efficiency of these antibiotics, notably azithromycin and clarithromycin, was 30% in most cases. CONCLUSION: The highest number of antibiotic-resistant bacteria, with a predominance of coliforms, was detected in samples of effluent biofilm. Multidrug-resistant strains in effluent biofilm showed very good biofilm-forming ability.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Azithromycin pharmacology MeSH
- Biofilms drug effects growth & development MeSH
- Enterobacteriaceae drug effects genetics isolation & purification MeSH
- Enterococcus drug effects genetics isolation & purification MeSH
- Clarithromycin pharmacology MeSH
- Wastewater microbiology MeSH
- Prevalence MeSH
- RNA, Ribosomal, 16S genetics MeSH
- Staphylococcus drug effects genetics isolation & purification MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Slovakia 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
- Anti-Bacterial Agents pharmacology MeSH
- Drug Resistance, Bacterial MeSH
- Bacterial Proteins genetics metabolism MeSH
- Enterococcus drug effects genetics isolation & purification physiology MeSH
- Virulence Factors genetics metabolism MeSH
- Microbial Sensitivity Tests MeSH
- Dogs microbiology MeSH
- Symbiosis MeSH
- Animals MeSH
- Check Tag
- Dogs microbiology MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
The present study focuses on the detection of enterococci in ostrich faeces. Forty-six bacterial colonies from 140 ostriches were identified at the species level using the MALDI-TOF MS identification system. According to the score value evaluation, they were allotted to the species Enterococcus hirae, Enterococcus faecium and Enterococcus mundtii confirmed also by phenotypic testing. Dominated species E. hirae (34 strains) were submitted to more detailed testing. Those strains E. hirae produced either no or only slight amount of the enzymes related to disorders (N-acetyl-β-glucosaminidase, β-glucuronidase, α-chymotrypsin, trypsin). Most of the strains were not hemolytic. They did not harbour the hiracin-producing gene. Five E. hirae strains harboured virulence factor gene gelE; however, they were phenotypically gelatinase negative. They also harboured other virulence factor genes such as esp, efaAfm and ccf. E. hirae strains were mostly sensitive to antibiotics and those resistant at least to one antibiotic were sensitive to enterocins (200-25,600 AU/mL). This study represents original and novel results concerning the enterococcal microflora in ostriches; enterococci in ostriches have not been described in detail up to now; sensitivity to enterocins of E. hirae strains harbouring virulence factor genes to enterocins is also new.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Enterococcus classification drug effects genetics isolation & purification MeSH
- Virulence Factors analysis genetics MeSH
- Feces microbiology MeSH
- Animals, Domestic MeSH
- Microbial Sensitivity Tests MeSH
- Bridged-Ring Compounds metabolism MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Struthioniformes microbiology MeSH
- Bacterial Typing Techniques MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Keywords
- avoparcin,
- MeSH
- Drug Resistance, Bacterial genetics drug effects MeSH
- Enterococcus physiology genetics classification ultrastructure MeSH
- Vancomycin-Resistant Enterococci * genetics drug effects ultrastructure MeSH
- Gene Expression genetics drug effects MeSH
- Glycopeptides drug effects MeSH
- Cross Infection mortality transmission MeSH
- Humans MeSH
- Vancomycin MeSH
- Zoonoses transmission MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
UNLABELLED: How bacteria control proper septum placement at midcell, to guarantee the generation of identical daughter cells, is still largely unknown. Although different systems involved in the selection of the division site have been described in selected species, these do not appear to be widely conserved. Here, we report that LocZ (Spr0334), a newly identified cell division protein, is involved in proper septum placement in Streptococcus pneumoniae. We show that locZ is not essential but that its deletion results in cell division defects and shape deformation, causing cells to divide asymmetrically and generate unequally sized, occasionally anucleated, daughter cells. LocZ has a unique localization profile. It arrives early at midcell, before FtsZ and FtsA, and leaves the septum early, apparently moving along with the equatorial rings that mark the future division sites. Consistently, cells lacking LocZ also show misplacement of the Z-ring, suggesting that it could act as a positive regulator to determine septum placement. LocZ was identified as a substrate of the Ser/Thr protein kinase StkP, which regulates cell division in S. pneumoniae. Interestingly, homologues of LocZ are found only in streptococci, lactococci, and enterococci, indicating that this close phylogenetically related group of bacteria evolved a specific solution to spatially regulate cell division. IMPORTANCE: Bacterial cell division is a highly ordered process regulated in time and space. Recently, we reported that the Ser/Thr protein kinase StkP regulates cell division in Streptococcus pneumoniae, through phosphorylation of several key proteins. Here, we characterized one of the StkP substrates, Spr0334, which we named LocZ. We show that LocZ is a new cell division protein important for proper septum placement and likely functions as a marker of the cell division site. Consistently, LocZ supports proper Z-ring positioning at midcell. LocZ is conserved only among streptococci, lactococci, and enterococci, which lack homologues of the Min and nucleoid occlusion effectors, indicating that these bacteria adapted a unique mechanism to find their middle, reflecting their specific shape and symmetry.
- MeSH
- Cell Division * MeSH
- Gene Deletion MeSH
- Enterococcus genetics MeSH
- Lactococcus genetics MeSH
- Cell Cycle Proteins genetics metabolism MeSH
- Sequence Homology, Amino Acid MeSH
- Streptococcus pneumoniae cytology genetics physiology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Only limited information exists concerning the microbiota in beaver (Castor fiber). This study has been focused on the virulence factors genes detection in enterococci from beavers. In general, animals are not affected by enterococcal infections, but they can be a reservoir of, e.g. pathogenic strains. Moreover, detection of virulence factors genes in enterococci from beavers was never tested before. Free-living beavers (12), male and female (age 4-5 years) were caught in the north-east part of Poland. Sampling of lower gut and faeces was provided according to all ethical rules for animal handling. Samples were treated using a standard microbiological method. Pure bacterial colonies were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) identification system. Virulence factors genes-gelE (gelatinase), agg (aggregation), cylA (cytolysin A), efaAfs (adhesin Enterococcus faecalis), efaAfm (adhesin Enterococcus faecium) and esp (surface protein) were tested by PCR. Moreover, gelatinase and antibiotic phenotypes were tested. Species detected were Enterococcus thailandicus, E. faecium, E. faecalis and Enterococcus durans. In literature, enterococcal species distribution was never reported yet up to now. Strains were mostly sensitive to antibiotics. Vancomycin-resistant E. faecalis EE9Tr1 possess cylA, efaAfs, esp and gelE genes. Strains were aggregation substance genes absent. Adhesin E. faecium (efaAfm) gene was detected in two of three E. faecium strains, but it was present also in E. thailandicus. Esp gene was present in EE9Tr1 and E. durans EDTr92. The most detected were gelE, efaAfm genes; in EF 4Hc1 also gelatinase phenotype was found. Strains with virulence factors genes will be tested for their sensitivity to antimicrobial enterocins.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Bacteriological Techniques MeSH
- Enterococcus classification drug effects genetics isolation & purification MeSH
- Virulence Factors analysis genetics MeSH
- Feces microbiology MeSH
- Genotype MeSH
- Gram-Positive Bacterial Infections microbiology veterinary MeSH
- Rodentia microbiology MeSH
- Microbial Sensitivity Tests MeSH
- Polymerase Chain Reaction MeSH
- Rectum microbiology MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Gelatinases analysis MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Poland MeSH
Coraciiform birds hoopoe (Upupa epops), common kingfisher (Alcedo atthis) and European roller (Coracius garrulus) were examined for enterococci in their cloacae and uropygial glands. The enterococcal isolates were identified at the species level using several genomic and proteomic methods, screened for antibiotic susceptibility and genotyped by pulsed-field gel electrophoresis (PFGE). Clonality of isolates from the common kingfisher was also assessed by multi-locus sequence typing (MLST). Using selective media, putative enterococcal isolates (n = 117) were recovered from 74% (32 out of a total of 43) of the bird samples and 114 isolates were confirmed as enterococci. Overall, among the total of 6 different species detected, Enterococcus faecalis was dominant (59%) in all three bird species. The second most frequently isolated species was Enterococcus casseliflavus (32%). PFGE revealed great diversity of strains from different bird species and anatomic location. Closely related strains were found only from nestlings from the same nest. No genes conferring resistance to vancomycin (vanA, vanB, vanC1 and van C2/C3) or erythromycin (erm A, ermB and mefA/E) were detected. MLST analysis and eBURST clustering revealed that sequence types of E. faecalis from the common kingfisher were identical to those of isolates found previously in water, chickens, and humans.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Biodiversity * MeSH
- Enterococcus classification drug effects genetics isolation & purification MeSH
- Phylogeny MeSH
- Microbial Sensitivity Tests MeSH
- Molecular Sequence Data MeSH
- Multilocus Sequence Typing MeSH
- Prevalence MeSH
- Birds microbiology MeSH
- Electrophoresis, Gel, Pulsed-Field MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
