The virulence factors, antibiotic resistance patterns, and the associated genetic elements have been investigated in Staphylococcus species. A total of 100 strains has been isolated from clinical samples in the Microbiology Laboratory of Hesperia Hospital, Modena, Italy, and identified as Staphylococcus aureus (65), Staphylococcus epidermidis (24), Staphylococcus hominis (3), Staphylococcus saprophyticus (3), and Staphylococcus warneri (5). All the strains were analyzed to determine phenotypic and genotypic characters, notably the virulence factors, the antibiotics susceptibility, and the genetic determinants. The highest percentage of resistance in Staphylococcus spp. was found for erythromycin and benzylpenicillin (87% and 85%, respectively). All S. aureus, two S. epidermidis (8.3%), and one S. saprophyticus (33.3%) strains were resistant to oxacillin. The methicillin resistance gene (mecA) was detected by polymerase chain reaction (PCR) amplification in 65 S. aureus strains and in 3 coagulase-negative staphylococci (CoNS) (8.6%). With regard to the virulence characteristics, all the S. aureus were positive to all virulence tests, except for slime test. Among the CoNS isolates, 19 (79.1%) S. epidermidis and one (33.3%) S. saprophyticus strains resulted positive for the slime test only. The results obtained are useful for a more in-depth understanding of the function and contribution of S. aureus and CoNS antibiotic resistance and virulence factors to staphylococcal infections. In particular, the production of slime is very important for CoNS, a virulence factor frequently found in infections caused by these strains. Further investigations on the genetic relatedness among strains of different sources will be useful for epidemiological and monitoring purposes and will enable us to develop new strategies to counteract the diffusion of methicillin-resistant S. aureus (MRSA) and CoNS strains not only in clinical field, but also in other related environments.
- MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Drug Resistance, Bacterial * MeSH
- Bacterial Proteins genetics MeSH
- Virulence Factors * genetics MeSH
- Humans MeSH
- Microbial Sensitivity Tests * MeSH
- Staphylococcal Infections * microbiology MeSH
- Staphylococcus * genetics drug effects isolation & purification pathogenicity classification MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Italy MeSH
- Keywords
- Staphylococcus brunensis,
- MeSH
- Humans MeSH
- Staphylococcus genetics isolation & purification MeSH
- Check Tag
- Humans MeSH
- Geographicals
- Czech Republic MeSH
Národní referenční laboratoř pro stafylokoky CEM–SZÚ se i v roce 2019 v rámci zajištění surveillance stafylokokových infekcí věnovala podrobnému vyšetřování kmenů stafylokoků z humánního klinického materiálu. Celkem to bylo 1405 kmenů, převážně druhu Staphylococcus aureus, které byly zaslány asi z 80 bakteriologických pracovišť z celé České republiky. Metodou PCR byla zjišťována přítomnost genů kódujících především Pantonův-Valentinův leukocidin, toxin Syndromu toxického šoku, exfoliatiny a enterotoxiny. Informace o produkci faktorů virulence jsou důležité pro ošetřujících lékaře ke správnému stanovení diagnózy a tedy i vhodné terapie. V celém souboru bylo i 79 (5,6 %) kmenů koaguláza negativních stafylokoků. U těchto podmíněných patogenů jsme fenotypizací a metodou hmotnostní spektrometrie kmeny identifikovali, resp. konfirmovali identifikaci zjištěnou již v terénních laboratořích.
The main focus of the National Reference Laboratory for Staphylococci (NRL) in 2019 was again on the investigation of staphylococcal strains from human clinical specimens within the surveillance of staphylococcal infections. In total, 1405 strains mostly of the species Staphylococcus aureus referred to the NRL by 80 bacteriological laboratories from all over the Czech Republic were analysed. The strains were screened by PCR for the genes encoding Panton- -Valentine leucocidin, toxic shock syndrome toxin, exfoliatins, and enterotoxins. Data on the production of virulence factors are helpful for attending physicians in determining the right diagnosis and effective treatment. Seventy-nine strains (5.6%) referred to the NRL were coagulase negative staphylococci. These opportunistic pathogens were identified or confirmed, after previous identification by field laboratories, by phenotyping and mass spectrometry.
- MeSH
- Enterotoxins isolation & purification classification MeSH
- Exotoxins isolation & purification MeSH
- Coagulase genetics isolation & purification MeSH
- Leukocidins isolation & purification MeSH
- Humans MeSH
- Polymerase Chain Reaction methods statistics & numerical data MeSH
- Shock, Septic epidemiology classification microbiology MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods instrumentation MeSH
- Staphylococcal Infections * epidemiology genetics classification MeSH
- Staphylococcus genetics isolation & purification classification pathogenicity MeSH
- Check Tag
- Humans MeSH
- Geographicals
- Czech Republic MeSH
Coagulase-negative staphylococci (CoNS) are an important cause of human and animal diseases. Treatment of these diseases is complicated by their common antimicrobial resistance, caused by overuse of antibiotics in hospital and veterinary environment. Therefore, they are assumed to serve as a reservoir of resistance genes often located on plasmids. In this study, we analyzed plasmid content in 62 strains belonging to 10 CoNS species of human and veterinary origin. In 48 (77%) strains analyzed, 107 different plasmids were detected, and only some of them showed similarities with plasmids found previously. In total, seven different antimicrobial-resistance genes carried by plasmids were identified. Five of the CoNS staphylococci carried plasmids identical with either those of other CoNS species tested, or a well characterized Staphylococcus aureus strain COL, suggesting plasmid dissemination through horizontal transfer. To demonstrate the possibility of horizontal transfer, we performed electroporation of four resistance plasmids among Staphylococcus epidermidis, Staphylococcus petrasii, and coagulase-positive S. aureus strains. Plasmids were transferred unchanged, were stably maintained in recipient strains, and expressed resistance genes. Our work demonstrates a great variability of plasmids in human and veterinary staphylococcal strains and their ability to maintain and express resistance plasmids from other staphylococcal species.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Drug Resistance, Bacterial genetics MeSH
- Genetic Variation * MeSH
- Coagulase MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Plasmids drug effects genetics MeSH
- Gene Transfer, Horizontal MeSH
- Staphylococcal Infections MeSH
- Staphylococcus aureus genetics MeSH
- Staphylococcus drug effects enzymology genetics MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
A group of 59 putative strains of Staphylococcus intermedius/Staphylococcus pseudintermedius deposited in the Czech National Collection of Type Cultures (CNCTC, National Institute for Public Health, Prague, Czech Republic) and the National Reference Laboratory for Staphylococci (NRL for Staphylococci, National Institute for Public Health, Prague, Czech Republic) was reclassified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). There the biggest human collection of S. pseudintermedius in Europe was analysed; 44 samples (75%) were of human origin. Twenty-two percent (n = 13) of the strains were isolated from animals, and two staphylococci were of unknown origin. This study revealed the prevalence of Staphylococcus pseudintermedius (94%, n = 53) vs. Staphylococcus intermedius (6%, n = 6) in the collection of human and veterinary staphylococci after reclassification. Results of PCR-RFLP analysis were verified by comparison with a repetitive element sequence-based polymerase chain reaction (Rep-PCR) analysis on 26 (44%) randomly selected strains. Due to a low-resolution ability of PCR-RFLP to separate Staphylococcus intermedius from Staphylococcus delphini, four isolates of Staphylococcus intermedius were biochemically verified further to exclude the presence of Staphylococcus delphini in the collection. Our results indicate that S. intermedius and S. pseudintermedius have occurred independently over an age-long period of their co-evolution.
- MeSH
- Bacterial Proteins genetics MeSH
- Bacteriological Techniques MeSH
- Biological Evolution * MeSH
- DNA, Bacterial genetics MeSH
- Humans MeSH
- Polymerase Chain Reaction MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Repetitive Sequences, Nucleic Acid genetics MeSH
- Sequence Analysis, DNA MeSH
- Staphylococcal Infections microbiology veterinary MeSH
- Staphylococcus intermedius classification genetics isolation & purification MeSH
- Staphylococcus classification genetics isolation & purification MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Staphylococcus petrasii is recently described coagulase negative staphylococcal species and an opportunistic human pathogen, still often misidentified in clinical specimens. Four subspecies are distinguished in S. petrasii by polyphasic taxonomical analyses, however a comparative study has still not been done on the majority of isolates and their genome properties have not yet been thoroughly analysed. Here, we describe the phenotypic and genotypic characteristics of 65 isolates and the results of de novo sequencing, whole genome assembly and annotation of draft genomes of five strains. The strains were identified by MALDI-TOF mass spectrometry to the species level and the majority of the strains were identified to the subspecies level by fingerprinting methods, (GTG)5 repetitive PCR and ribotyping. Macrorestriction profiling by pulsed-field gel electrophoresis was confirmed to be a suitable strain typing method. Comparative genomics revealed the presence of new mobile genetic elements carrying antimicrobial resistance factors such as staphylococcal cassette chromosome (SCC) mec, transposones, phage-inducible genomic islands, and plasmids. Their mosaic structure and similarity across coagulase-negative staphylococci and Staphylococcus aureus suggest the possible exchange of these elements. Numerous putative virulence factors such as adhesins, autolysins, exoenzymes, capsule formation genes, immunomodulators, the phage-associated sasX gene, and SCC-associated spermidine N-acetyltransferase gene, pseudouridine and sorbitol utilization operons might explain clinical manifestations of S. petrasii isolates. The increasing recovery of S. petrasii isolates from human clinical material, the multi-drug resistance including methicillin resistance of S. petrasii subsp. jettensis strains, and virulence factors homologous to other pathogenic staphylococci demonstrate the importance of the species in human disease.
- MeSH
- Virulence Factors genetics MeSH
- Phenotype MeSH
- Genome, Bacterial * MeSH
- Genomics MeSH
- Genotype MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Electrophoresis, Gel, Pulsed-Field MeSH
- Ribotyping MeSH
- Interspersed Repetitive Sequences * MeSH
- Staphylococcus classification genetics pathogenicity MeSH
- Bacterial Typing Techniques MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
The aim of this study was to assess pit latrine samples from a peri-urban community in KwaZulu-Natal (South Africa) for the presence of multidrug-resistant (MDR) Staphylococcus spp. Standard procedures were used to isolate Staphylococcus spp. from pit latrine fecal sludge samples, with confirmation at genus level by polymerase chain reaction (PCR). Sixty-eight randomly selected pit latrine Staphylococcus spp. isolates were further characterized by using established disk diffusion procedures. An average Staphylococcus spp. count of 2.1 × 105 CFU per g fecal material was established using two randomly selected pit latrine samples. Of the 68-selected Staphylococcus spp. pit latrine isolates, 49% were identified as coagulase positive, 51% as coagulase negative and 65% (12 coagulase positive, 32 coagulase negative isolates) were categorized as MDR. The majority (66/68) of Staphylococcus spp. isolates displayed resistance to fusidic acid while only 5/68 isolates displayed resistance to chloramphenicol. The pit latrine samples analyzed in this study are a source of MDR Staphylococcus spp., highlighting the need for proper hygiene and sanitation regimes in rural communities using these facilities.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Genes, Bacterial genetics MeSH
- Bacterial Proteins metabolism MeSH
- DNA, Bacterial genetics MeSH
- Feces microbiology MeSH
- Coagulase metabolism MeSH
- Microbial Sensitivity Tests MeSH
- Microbial Viability drug effects MeSH
- Drug Resistance, Multiple, Bacterial * MeSH
- Sewage microbiology MeSH
- Staphylococcus drug effects enzymology genetics isolation & purification MeSH
- Toilet Facilities * MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
- Geographicals
- South Africa MeSH
- MeSH
- Bacterial Proteins genetics MeSH
- Glycopeptides pharmacology MeSH
- Polymorphism, Single Nucleotide MeSH
- Humans MeSH
- Membrane Proteins genetics MeSH
- Microbial Sensitivity Tests MeSH
- Mutation MeSH
- Staphylococcal Infections microbiology MeSH
- Staphylococcus drug effects genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Letter MeSH
- Research Support, Non-U.S. Gov't MeSH
Vagina which is one of the important reservoirs for Staphylococcus and in pregnant women pathogenic strains may infect the child during the birth or by vertical transmission. A total of 68 presumptive Staphylococcus strains isolated from human vagina were found to be gram-positive cocci, and only 32 (47%) isolates were found beta-hemolytic. Matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) results confirmed 33 isolates belonged to Staphylococcus which consisting of 6 species, i.e., S. aureus (14), S. vitulinus (7), S. epidermidis (4), S cohnii (3), S. equorum (3), and S. succinus (2). Further, the result of antibiotic susceptibility tests showed that large proportions (76%-100%) of the isolates were resistant to multiple antibiotics and more often resistant to penicillin (100%), ampicillin (100%), oxacillin (97%), oxytetracycline (97%), vancomycin (97%), rifampin (85%), erythromycin (82%), and streptomycin (76%). In the present study, only the sec enterotoxin gene was detected in four S. aureus strains. DNA fingerprints of the 33 isolates that were generated using random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) PCR analysis revealed great genetic relatedness of isolates. High prevalence of vaginal colonization with multiple antibiotic-resistant staphylococci among pregnant women was observed which were emerged from the single respective species clones that underwent evolution. The vertical transmission of these multiple antibiotic-resistant Staphylococcus species to the infant is possible; therefore, the findings of this study emphasize the need for regular surveillance of antibiotic-resistant bacterial strains in pregnant women in this area.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Genes, Bacterial genetics MeSH
- DNA, Bacterial genetics MeSH
- Enterotoxins genetics MeSH
- Virulence Factors genetics MeSH
- Genotype MeSH
- Pregnancy Complications, Infectious microbiology MeSH
- Humans MeSH
- Drug Resistance, Multiple, Bacterial * MeSH
- Cluster Analysis MeSH
- Staphylococcal Infections microbiology MeSH
- Staphylococcus classification genetics isolation & purification MeSH
- Pregnancy MeSH
- Vagina microbiology MeSH
- Check Tag
- Humans MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Geographicals
- South Africa 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
