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PloS One 2016Staphylococcus haemolyticus is one of the most common pathogens associated with medical-device related infections, but its molecular epidemiology is poorly explored. In...
Staphylococcus haemolyticus is one of the most common pathogens associated with medical-device related infections, but its molecular epidemiology is poorly explored. In the current study, we aimed to better understand the genetic mechanisms contributing to S. haemolyticus diversity in the hospital environment and their impact on the population structure and clinical relevant phenotypic traits. The analysis of a representative S. haemolyticus collection by multilocus sequence typing (MLST) has identified a single highly prevalent and diverse genetic lineage of nosocomial S. haemolyticus clonal complex (CC) 29 accounting for 91% of the collection of isolates disseminated worldwide. The examination of the sequence changes at MLST loci during clonal diversification showed that recombination had a higher impact than mutation in shaping the S. haemolyticus population. Also, we ascertained that another mechanism contributing significantly to clonal diversification and adaptation was mediated by insertion sequence (IS) elements. We found that all nosocomial S. haemolyticus, belonging to different STs, were rich in IS1272 copies, as determined by Southern hybridization of macrorestriction patterns. In particular, we observed that the chromosome of a S. haemolyticus strain within CC29 was highly unstable during serial growth in vitro which paralleled with IS1272 transposition events and changes in clinically relevant phenotypic traits namely, mannitol fermentation, susceptibility to beta-lactams, biofilm formation and hemolysis. Our results suggest that recombination and IS transposition might be a strategy of adaptation, evolution and pathogenicity of the major S. haemolyticus prevalent lineage in the hospital environment.
Topics: Anti-Bacterial Agents; Electrophoresis, Gel, Pulsed-Field; Genes, Bacterial; Microbial Sensitivity Tests; Mutation; Phylogeny; Recombination, Genetic; Staphylococcus haemolyticus
PubMed: 27249649
DOI: 10.1371/journal.pone.0156653 -
Memorias Do Instituto Oswaldo Cruz Mar 2017The association between Staphylococcus haemolyticus and severe nosocomial infections is increasing. However, the extent to which fomites contribute to the dissemination...
BACKGROUND
The association between Staphylococcus haemolyticus and severe nosocomial infections is increasing. However, the extent to which fomites contribute to the dissemination of this pathogen through patients and hospital wards remains unknown.
OBJECTIVES
In the present study, sphygmomanometers and thermometers were evaluated as potential fomites of oxacillin-resistant S. haemolyticus (ORSH). The influence of oxacillin and vancomycin on biofilm formation by ORSH strains isolated from fomites was also investigated.
METHODS
The presence of ORSH on swabs taken from fomite surfaces in a Brazilian hospital was assessed using standard microbiological procedures. Antibiotic susceptibility profiles were determined by the disk diffusion method, and clonal distribution was assessed in pulsed-field gel electrophoresis (PFGE) assays. Minimum inhibitory concentrations (MICs) of oxacillin and vancomycin were evaluated via the broth microdilution method. Polymerase chain reaction (PCR) assays were performed to detect the mecA and icaAD genes. ORSH strains grown in media containing 1/4 MIC of vancomycin or oxacillin were investigated for slime production and biofilm formation on glass, polystyrene and polyurethane catheter surfaces.
FINDINGS
ORSH strains comprising five distinct PFGE types were isolated from sphygmomanometers (n = 5) and a thermometer (n = 1) used in intensive care units and surgical wards. ORSH strains isolated from fomites showed susceptibility to only linezolid and vancomycin and were characterised as multi-drug resistant (MDR). Slime production, biofilm formation and the survival of sessile bacteria differed and were independent of the presence of the icaAD and mecA genes, PFGE type and subtype. Vancomycin and oxacillin did not inhibit biofilm formation by vancomycin-susceptible ORSH strains on abiotic surfaces, including on the catheter surface. Enhanced biofilm formation was observed in some situations. Moreover, a sub-lethal dose of vancomycin induced biofilm formation by an ORSH strain on polystyrene.
MAIN CONCLUSIONS
Sphygmomanometers and thermometers are fomites for the transmission of ORSH. A sub-lethal dose of vancomycin may favor biofilm formation by ORSH on fomites and catheter surfaces.
Topics: Anti-Bacterial Agents; Biofilms; Cross Infection; Drug Resistance, Bacterial; Electrophoresis, Gel, Pulsed-Field; Fomites; Humans; Microbial Sensitivity Tests; Oxacillin; Sphygmomanometers; Staphylococcal Infections; Staphylococcus haemolyticus; Thermometers; Vancomycin
PubMed: 28225903
DOI: 10.1590/0074-02760160381 -
Journal of Dairy Science Sep 2014The aim of this review is to assess the effect of coagulase-negative staphylococci (CNS) species on udder health and milk yield in ruminants, and to evaluate the... (Review)
Review
The aim of this review is to assess the effect of coagulase-negative staphylococci (CNS) species on udder health and milk yield in ruminants, and to evaluate the capacity of CNS to cause persistent intramammary infections (IMI). Furthermore, the literature on factors suspected of playing a role in the pathogenicity of IMI-associated CNS, such as biofilm formation and the presence of various putative virulence genes, is discussed. The focus is on the 5 CNS species that have been most frequently identified as causing bovine IMI using reliable molecular identification methods (Staphylococcus chromogenes, Staphylococcus simulans, Staphylococcus haemolyticus, Staphylococcus xylosus, and Staphylococcus epidermidis). Although the effect on somatic cell count and milk production is accepted to be generally limited or nonexistent for CNS as a group, indications are that the typical effects differ between CNS species and perhaps even strains. It has also become clear that many CNS species can cause persistent IMI, contrary to what has long been believed. However, this trait appears to be quite complicated, being partly strain dependent and partly dependent on the host's immunity. Consistent definitions of persistence and more uniform methods for testing this phenomenon will benefit future research. The factors explaining the anticipated differences in pathogenic behavior appear to be more difficult to evaluate. Biofilm formation and the presence of various staphylococcal virulence factors do not seem to (directly) influence the effect of CNS on IMI but the available information is indirect or insufficient to draw consistent conclusions. Future studies on the effect, persistence, and virulence of the different CNS species associated with IMI would benefit from using larger and perhaps even shared strain collections and from adjusting study designs to a common framework, as the large variation currently existing therein is a major problem. Also within-species variation should be investigated.
Topics: Animals; Biofilms; Cattle; Coagulase; Female; Mammary Glands, Animal; Mastitis, Bovine; Milk; Phenotype; Species Specificity; Staphylococcal Infections; Staphylococcus; Virulence Factors
PubMed: 24952781
DOI: 10.3168/jds.2013-7775 -
Microbial Genomics Jan 2022is a species of coagulase-negative staphylococci that has primarily been studied as a human skin microbiome member and an emerging nosocomial pathogen. Here, we present...
is a species of coagulase-negative staphylococci that has primarily been studied as a human skin microbiome member and an emerging nosocomial pathogen. Here, we present the first complete genome of strains SE3.9, SE3.8 and SE2.14 reported as an endophyte of rice seed. Detailed investigation of the genome dynamics of strains from diverse origins revealed an expanded genome size in clinical isolates, and a role of many insertion sequence (IS) elements in strain diversification. Interestingly, several of the IS elements are also unique or enriched in a particular habitat. Comparative studies also revealed the potential movement of mobile elements from rice endophytic to strains from other pathogenic species such as . The study highlights the importance of ecological studies in the systematic understanding of genome plasticity and management of medically important species.
Topics: DNA Transposable Elements; Genome Size; Genome, Bacterial; High-Throughput Nucleotide Sequencing; Oryza; Seeds; Staphylococcus haemolyticus; Whole Genome Sequencing
PubMed: 35072602
DOI: 10.1099/mgen.0.000755 -
BMC Microbiology Sep 2023Staphylococcus haemolyticus (S. haemolyticus) is the main etiological factor in skin and soft tissue infections (SSTI). S. haemolyticus infections are an important...
BACKGROUND
Staphylococcus haemolyticus (S. haemolyticus) is the main etiological factor in skin and soft tissue infections (SSTI). S. haemolyticus infections are an important concern worldwide, especially with the associated biofilms and drug resistance. Herein, we investigated the inhibitory effect of Flavaspidic acid BB obtained from plant extractions on clinical S. haemolyticus strains and their biofilms. Moreover, we predicted its ability to bind to the protein-binding site by molecular simulation. Since the combination of Hsp70 and RNase P synthase after molecular simulation with flavaspidic acid BB is relatively stable, enzyme-linked immunosorbent assay (ELISA) was used to investigate Hsp70 and RNase P synthase to verify the potential antimicrobial targets of flavaspidic acid BB.
RESULTS
The minimum inhibitory concentrations (MIC) of flavaspidic acid BB on 16 clinical strains of S. haemolyticus was 5 ~ 480 µg/mL, and BB had a slightly higher inhibitory effect on the biofilm than MUP. The inhibitory effect of flavaspidic acid BB on biofilm formation was better with an increase in the concentration of BB. Molecular simulation verified its ability to bind to the protein-binding site. The combination of ELISA kits showed that flavaspidic acid BB promoted the activity of Hsp70 and inhibited the activity of RNase P, revealing that flavaspidic acid BB could effectively inhibit the utilization and re-synthesis of protein and tRNA synthesis, thus inhibiting bacterial growth and biofilm formation to a certain extent.
CONCLUSIONS
This study could potentially provide a new prospect for the development of flavaspidic acid BB as an antibacterial agent for resistant strains.
Topics: Staphylococcus; Ribonuclease P; Anti-Bacterial Agents; Butyrophenones; Microbial Sensitivity Tests; Biofilms
PubMed: 37773054
DOI: 10.1186/s12866-023-02997-5 -
Annals of Clinical Microbiology and... May 2017The objectives of this study were to characterize the diversity and magnitude of antimicrobial resistance among Staphylococcus species recovered from imported beef meat...
BACKGROUND
The objectives of this study were to characterize the diversity and magnitude of antimicrobial resistance among Staphylococcus species recovered from imported beef meat sold in the Egyptian market and the potential mechanisms underlying the antimicrobial resistance phenotypes including harboring of resistance genes (mecA, cfr, gyrA, gyrB, and grlA) and biofilm formation.
RESULTS
The resistance gene mecA was detected in 50% of methicillin-resistant non-Staphylococcus aureus isolates (4/8). Interestingly, our results showed that: (i) resistance genes mecA, gyrA, gyrB, grlA, and cfr were absent in Staphylococcus hominis and Staphylococcus hemolyticus isolates, although S. hominis was phenotypically resistant to methicillin (MR-non-S. aureus) while S. hemolyticus was resistant to vancomycin only; (ii) S. aureus isolates did not carry the mecA gene (100%) and were phenotypically characterized as methicillin- susceptible S. aureus (MSS); and (iii) the resistance gene mecA was present in one isolate (1/3) of Staphylococcus lugdunensis that was phenotypically characterized as methicillin-susceptible non-S. aureus (MSNSA).
CONCLUSIONS
Our findings highlight the potential risk for consumers, in the absence of actionable risk management information systems, of imported foods and advice a strict implementation of international standards by different venues such as CODEX to avoid the increase in prevalence of coagulase positive and coagulase negative Staphylococcus isolates and their antibiotic resistance genes in imported beef meat at the Egyptian market.
Topics: Animals; Anti-Bacterial Agents; Bacterial Proteins; Biofilms; Cattle; Chlorocebus aethiops; Coagulase; DNA Gyrase; Drug Resistance, Bacterial; Egypt; Food Microbiology; Genes, Bacterial; Hemolysin Proteins; Methicillin; Methicillin Resistance; Microbial Sensitivity Tests; Penicillin-Binding Proteins; Phenotype; RNA, Ribosomal, 16S; Red Meat; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus; Staphylococcus haemolyticus; Staphylococcus lugdunensis; Vancomycin; Vero Cells; Virulence
PubMed: 28486995
DOI: 10.1186/s12941-017-0210-4 -
Central European Journal of Public... Jun 2022This work aimed to determine the representation and resistance of bacteria belonging to the genus Staphylococcus and Enterococcus on inanimate surfaces of two selected...
OBJECTIVES
This work aimed to determine the representation and resistance of bacteria belonging to the genus Staphylococcus and Enterococcus on inanimate surfaces of two selected workplaces of the University Hospital of L. Pasteur in Košice (UHLP) and to investigate their importance in the hospital environment. The men's ward of the Department of Internal Medicine (DIM) and the Department of Anaesthesiology and Intensive Care (DAIC) were chosen.
METHODS
Using sterile sampling kits, a total of 182 swabs were collected from the inanimate surfaces of both UHLP workplaces. The swabs were then transported to a microbiological laboratory and inoculated onto sterile culture media (blood agar containing 5% ram erythrocytes). After culturing (24-48 hours, in a thermostat at constant temperature 37 °C), bacterial colonies were identified by mass spectrometry on a MALDI TOF MS. Bacteria belonging to the genera Staphylococcus and Enterococcus were subsequently separated from the spectrum of identified bacteria. Nosocomial significant strains of staphylococci (Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus aureus) and all isolated enterococci were subjected to susceptibility testing for selected antibiotics using the disk diffusion method - E-tests.
RESULTS
Several members of the genus Staphylococcus were identified from the inanimate surfaces of both workplaces. These were mainly coagulase-negative strains - Staphylococcus epidermidis (45), Staphylococcus capitis (34), Staphylococcus haemolyticus (20), Staphylococcus hominis (45), Staphylococcus pasteuri (2), Staphylococcus sroph (1), Staphylococcus simulans (3), and Staphylococcus warneri (4). Staphylococcus aureus strains were also identified (2). Nosocomial significant isolates were tested for susceptibility to the antibiotics cefoxitin (FOX) and oxacillin (OXA). Two members of the genus Enterococcus - Enterococcus faecium (7) and Enterococcus faecalis (8) were isolated. All strains were subject to vancomycin susceptibility testing using the disk method.
Topics: Animals; Anti-Bacterial Agents; Bacteria; Cross Infection; Enterococcus; Hospitals; Humans; Male; Microbial Sensitivity Tests; Sheep; Staphylococcal Infections; Staphylococcus
PubMed: 35841227
DOI: 10.21101/cejph.a7241 -
Infection, Genetics and Evolution :... Jan 2022The surface protein SasX, has a key role in methicillin-resistant Staphylococcus aureus (MRSA) colonization and pathogenesis, and has been associated with the epidemic...
The surface protein SasX, has a key role in methicillin-resistant Staphylococcus aureus (MRSA) colonization and pathogenesis, and has been associated with the epidemic success of some MRSA clones. To date, only one SasX homologous protein, named SesI, has been described in Staphylococcus epidermidis. In this work, we analyze the occurrence of the sasX gene and its genetic environment in Staphylococcus haemolyticus S. haemolyticus clinical strains (n = 62) were screened for the presence of the sasX gene and its carrier, the prophage Φ SPβ-like. A deep characterization was done in one strain (MD43), through which we determined the complete nucleotide sequence for the S. haemolitycus sasX-like gene. Whole genome sequencing of strain MD43 was performed, and the gene, termed here because of its unique attributes, shsA, was mapped to the Φ SPβ-like prophage sequence. The shsA gene was detected in 33 out of 62 strains showing an average identity of 92 and 96% with the sasX and sesI genes and at the amino acid level, 88% identity with SasX and 92% identity with SesI. The ~124Kb Φ SPβ-like prophage sequence showed a largely intact prophage compared to its counterpart in S. epidermidis strain RP62A, including the sesI insertion site. In conclusion, we identified a new sasX ortholog in S. haemolyticus (shsA). Its horizontal spread from this reservoir could represent an emergent threat in healthcare facilities since so far, no S. aureus sasX strains have been reported in Brazil.
Topics: Brazil; Genome, Bacterial; Prophages; Staphylococcus haemolyticus; Virulence; Virulence Factors; Whole Genome Sequencing
PubMed: 34920103
DOI: 10.1016/j.meegid.2021.105189 -
Microbiological Research Mar 2010In this study, we standardized and evaluated a multiplex-PCR methodology using specific primers to identify Staphylococcus aureus, Staphylococcus epidermidis and...
Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus: methicillin-resistant isolates are detected directly in blood cultures by multiplex PCR.
In this study, we standardized and evaluated a multiplex-PCR methodology using specific primers to identify Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus and their methicillin-resistance directly from blood cultures. Staphylococci clinical isolates (149) and control strains (16) previously identified by conventional methods were used to establish the multiplex PCR protocol. Subsequently, this methodology was evaluated using a fast and cheap DNA extraction protocol from 25 staphylococci positive blood cultures. A wash step of the pellet with 0.1% bovine serum albumin (BSA) solution was performed to reduce PCR inhibitors. Amplicons of 154bp (mecA gene), 271bp (S. haemolyticus mvaA gene) and 108 and 124bp (S. aureus and S. epidermidis species-specific fragments, respectively) were observed. Reliable results were obtained for 100% of the evaluated strains, suggesting that this new multiplex-PCR combined with an appropriate DNA-extraction method could be useful in the laboratory for fast and accurate identification of three staphylococci species and simultaneously their methicillin resistance directly in blood cultures.
Topics: Bacteriological Techniques; Blood; DNA Primers; Humans; Methicillin Resistance; Polymerase Chain Reaction; Sensitivity and Specificity; Staphylococcal Infections; Staphylococcus aureus; Staphylococcus epidermidis; Staphylococcus haemolyticus
PubMed: 19616418
DOI: 10.1016/j.micres.2009.03.003 -
The Review of Diabetic Studies : RDS Jun 2022Our objective was to assess the pattern of urine infections, the most common pathogen, and their susceptibility pattern to antibiotics among Saudi diabetic patients....
Our objective was to assess the pattern of urine infections, the most common pathogen, and their susceptibility pattern to antibiotics among Saudi diabetic patients. We performed a year-long cross-sectional study from January 2018 to January 2019 at KAAU Hospital in Riyadh, KSA. We cultured the urine specimens obtained from diabetic patients based on optimal aerobic and anaerobic microbiological methods. By adopting standard microbiological methods, we identified the bacterial isolates. We also followed the guidelines of the Clinical and Laboratory Standards Institute (CLSI) to do antibiotic susceptibility testing. A total of 100 isolates were evaluated, and a total of 22 organisms were isolated. The majority were multidrug-resistant organisms. was the most frequent organism and rated (15%). It was followed by (11%), (9%), (9%), (7%), (7%), (7%), (5%) and (5%). We also found multi-microbial infections. Most of the organisms were susceptible to tigecycline, gentamycin, and nitrofurantoin, rating (88%), (84%) and (78%), respectively. Our study revealed that a wide range of pathogens affects the diabetes patients. is the most prevalent pathogen. We observed considerable antimicrobial resistance. Tigecycline had a wide sensitivity spectrum and was effective against most of the bacteria. Thus, it can be used as an empirical antibiotic.
Topics: Anti-Bacterial Agents; Bacterial Infections; Cross-Sectional Studies; Diabetes Mellitus; Enterococcus; Escherichia coli; Gram-Negative Bacteria; Gram-Positive Bacteria; Humans; Microbial Sensitivity Tests; Tigecycline
PubMed: 35831935
DOI: 10.1900/RDS.2022.18.135