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1.
Microb Drug Resist ; 26(3): 251-260, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31549905

ABSTRACT

This study compared changes in antimicrobial susceptibilities and molecular characteristics of coagulase-negative staphylococci (CNS) between the year 2000 and the year 2014-2015 to evaluate the policy of separating drug prescribing and dispensing in Korea. We obtained 68 CNS clinical isolates from two tertiary general hospitals before (the year 2000; n = 25) and after (the year 2014 - 2015; n = 43) implementation of the separation. Isolates were identified as Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus, and Staphylococcus warneri. When minimal inhibitory concentrations of 14 antimicrobials were applied to isolates, resistance rates to gentamicin and oxacillin in 2000 were significantly higher than in 2014-2015 (p < 0.05). Fifty-seven isolates were methicillin-resistant CNS (MR-CNS), 42 of which were also multidrug resistant; overall, multidrug resistance decreased from 72% in the year 2000 to 55.8% in 2014-2015. Staphylococcal cassette chromosome mec (SCCmec) type III was the dominant type of MR-CNS in the year 2000, while SCCmec type IV was the dominant type in 2014-2015. Twenty-five sequence types (STs) were identified; ST2 appeared most frequently in both periods. After 15 years of implementation of this policy, multidrug resistance as well as methicillin and gentamicin resistance in CNS decreased, but not resistance to other antibiotics. Long-term surveillance at both genotypic and phenotypic levels of various species is necessary for further evaluation of this policy.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Prescriptions/statistics & numerical data , Drug Resistance, Multiple, Bacterial/genetics , Staphylococcal Infections/epidemiology , Staphylococcus epidermidis/genetics , Staphylococcus haemolyticus/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Coagulase/deficiency , Coagulase/genetics , Gene Expression , Gentamicins/pharmacology , Humans , Legislation, Drug , Microbial Sensitivity Tests , Oxacillin/pharmacology , Phylogeny , Republic of Korea/epidemiology , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus , Staphylococcus capitis/classification , Staphylococcus capitis/drug effects , Staphylococcus capitis/genetics , Staphylococcus capitis/isolation & purification , Staphylococcus epidermidis/classification , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/isolation & purification , Staphylococcus haemolyticus/classification , Staphylococcus haemolyticus/drug effects , Staphylococcus haemolyticus/isolation & purification , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Staphylococcus hominis/isolation & purification , Staphylococcus saprophyticus , Tertiary Care Centers
2.
Sci Rep ; 9(1): 11736, 2019 08 13.
Article in English | MEDLINE | ID: mdl-31409863

ABSTRACT

Staphylococcus hominis is part of the normal human microbiome. Two subspecies, S. hominis hominis (Shh) and S. hominis novobiosepticus (Shn), have clinical significance. Forty-nine S. hominis isolates were analyzed by the MicroScan automated system, SDS-PAGE and MALDI-TOF methods, followed by partial sequencing of the 16S rDNA gene. The trehalose fermentation test, disk diffusion and broth microdilution tests were used to identify (novobiocin test) and access the susceptibility to oxacillin and vancomycin of isolates. The SCCmec elements and genomic diversity were evaluated by PCR and PFGE methods, respectively. Profiles of 28 (57%; 8 Shh and 20 Shn) isolates corroborated with the results found in all the applied methods of identification. The remaining 21 (43%) isolates were phenotypically identified as Shh by MicroScan; however, they were identified as Shn by SDS-PAGE and mass spectral, and confirmed by 16S rDNA sequencing. Among 41 isolates identified as Shn by the molecular and mass spectrometry methods, 19 (41%) were novobiocin-sensitive, and the trehalose test indicated 11 positive isolates, which are considered atypical phenotypic results for this subspecies. In addition, 92.7% of the isolates identified as Shn by these methods carried mecA gene, while only 12.5% of the Shh isolates were positive. Together, the results highlighted the SDS-PAGE and MALDI-TOF MS methods as promising tools for discriminating S. hominis subspecies.


Subject(s)
Bacterial Proteins/metabolism , Electrophoresis, Polyacrylamide Gel , Proteome , Proteomics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Staphylococcus hominis/classification , Staphylococcus hominis/metabolism , Humans , Proteomics/methods
3.
J Basic Microbiol ; 59(5): 477-486, 2019 May.
Article in English | MEDLINE | ID: mdl-30900761

ABSTRACT

In the present study, a lead (Pb)-resistant bacterium, Staphylococcus hominis strain AMB-2 was isolated from Mandoli industrial area, Delhi and selected for heavy metal biosorption considering multiple heavy metal resistance. In the batch experiment, both living and dead biomasses of strain AMB-2 showed biosorption of Pb and cadmium (Cd) in single and binary systems as analyzed through Inductively coupled plasma-optical emission spectrometry. Living biomass exhibited more biosorption of metals than dead biomass in both single and binary systems. However, in the binary system, metals competed for the attachment sites on the bacterial surface, where Pb got more preference over Cd for the same. The underlying mechanism for the biosorption was attachment of the metal ions through functional groups onto the surface of the biomass as revealed by scanning electron microscope-energy-dispersive X-ray spectroscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction. Conclusively, this study displayed an effective biotreatment of Pb and Cd from aqueous medium using a low-cost biosorbent prepared from S. hominis strain AMB-2 considering biosafety of microorganisms and an eco-friendly approach.


Subject(s)
Lead/metabolism , Metals, Heavy/metabolism , Staphylococcus hominis/metabolism , Water Pollutants, Chemical/metabolism , Adsorption , Biodegradation, Environmental , Biomass , Cadmium/metabolism , Microbial Sensitivity Tests , Phylogeny , Soil Microbiology , Staphylococcus hominis/classification , Staphylococcus hominis/isolation & purification , Staphylococcus hominis/ultrastructure
4.
Microbiome ; 6(1): 213, 2018 11 29.
Article in English | MEDLINE | ID: mdl-30497517

ABSTRACT

BACKGROUND: Even though human sweat is odorless, bacterial growth and decomposition of specific odor precursors in it is believed to give rise to body odor in humans. While mechanisms of odor generation have been widely studied in adults, little is known for teenagers and pre-pubescent children who have distinct sweat composition from immature apocrine and sebaceous glands, but are arguably more susceptible to the social and psychological impact of malodor. RESULTS: We integrated information from whole microbiome analysis of multiple skin sites (underarm, neck, and head) and multiple time points (1 h and 8 h after bath), analyzing 180 samples in total to perform the largest metagenome-wide association study to date on malodor. Significant positive correlations were observed between odor intensity and the relative abundance of Staphylococcus hominis, Staphylococcus epidermidis, and Cutibacterium avidum, as well as negative correlation with Acinetobacter schindleri and Cutibacterium species. Metabolic pathway analysis highlighted the association of isovaleric and acetic acid production (sour odor) from enriched S. epidermidis (teen underarm) and S. hominis (child neck) enzymes and sulfur production from Staphylococcus species (teen underarm) with odor intensity, in good agreement with observed odor characteristics in pre-pubescent children and teenagers. Experiments with cultures on human and artificial sweat confirmed the ability of S. hominis and S. epidermidis to independently produce malodor with distinct odor characteristics. CONCLUSIONS: These results showcase the power of skin metagenomics to study host-microbial co-metabolic interactions, identifying distinct pathways for odor generation from sweat in pre-pubescent children and teenagers and highlighting key enzymatic targets for intervention.


Subject(s)
Bacteria/classification , Metagenomics/methods , Odorants/analysis , Skin/microbiology , Sweat/microbiology , Acetic Acid/analysis , Acinetobacter/classification , Acinetobacter/isolation & purification , Adolescent , Axilla/microbiology , Bacteria/isolation & purification , Child , Female , Head/microbiology , Hemiterpenes , Humans , Male , Neck/microbiology , Pentanoic Acids/analysis , Propionibacteriaceae/classification , Propionibacteriaceae/isolation & purification , Puberty , Sequence Analysis, DNA , Skin/chemistry , Staphylococcus epidermidis/classification , Staphylococcus epidermidis/isolation & purification , Staphylococcus hominis/classification , Staphylococcus hominis/isolation & purification , Sulfur/analysis
5.
Eur J Clin Microbiol Infect Dis ; 37(8): 1539-1545, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29777490

ABSTRACT

The study was performed to assess potential differences in the etiological relevance of two coagulase-negative staphylococci (CoNS), Staphylococcus haemolyticus and Staphylococcus hominis, in an observational single-center study. Over a 5-year interval, patients in whom there was detected S. haemolyticus or S. hominis of presumed etiological relevance were assessed for the primary endpoint death during hospital stay and the secondary endpoint transfer to an intensive care unit (ICU) after the detection of S. haemolyticus or S. hominis. Patients with S. haemolyticus or S. hominis died in 11.3% (50 out of 444) and 9.5% (60 out of 631) of cases, respectively, and were transferred to ICU after S. haemolyticus and S. hominis detection in 8.7% (19 out of 219) and 11.7% (44 out of 377) of cases, respectively. There was no significance for species-related influence on the primary outcome parameter (P > 0.1), while ICU transfers were more likely for patients with S. hominis detections (P = 0.016). Delayed diagnosis of both CoNS species was associated with an increased probability of death (P = 0.009). The study revealed comparable morbidity caused by S. haemolyticus and S. hominis identified in a clinically relevant context.


Subject(s)
Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus haemolyticus , Staphylococcus hominis , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Bacterial Typing Techniques , Biodiversity , Coagulase/genetics , Germany/epidemiology , Humans , Incidence , Intensive Care Units , Length of Stay , Outcome Assessment, Health Care , Retrospective Studies , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Staphylococcal Infections/diagnosis , Staphylococcal Infections/drug therapy , Staphylococcus haemolyticus/classification , Staphylococcus haemolyticus/drug effects , Staphylococcus haemolyticus/genetics , Staphylococcus haemolyticus/isolation & purification , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Staphylococcus hominis/isolation & purification
6.
Folia Microbiol (Praha) ; 61(2): 143-7, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26253583

ABSTRACT

Coagulase-negative staphylococci (CoNS) are the most frequently isolated bacteria from the blood and the predominant cause of nosocomial infections. Macrolides, lincosamides and streptogramin B (MLSB) antibiotics, especially erythromycin and clindamycin, are important therapeutic agents in the treatment of methicillin-resistant staphylococci infections. Among CoNS, Staphylococcus hominis represents the third most common organism. In spite of its clinical significance, very little is known about its mechanisms of resistance to antibiotics, especially MLSB. Fifty-five S. hominis isolates from the blood and the surgical wounds of hospitalized patients were studied. The erm(C) gene was predominant in erythromycin-resistant S. hominis isolates. The methylase genes, erm(A) and erm(B), were present in 15 and 25% of clinical isolates, respectively. A combination of various erythromycin resistance methylase (erm) genes was detected in 15% S. hominis isolates. The efflux gene msr(A) was detected in 18% of isolates, alone in four isolates, and in different combinations in a further six. The lnu(A) gene, responsible for enzymatic inactivation of lincosamides was carried by 31% of the isolates. No erythromycin resistance that could not be attributed to the genes erm(A), erm(B), erm(C) and msr(A) was detected. In S. hominis, 75 and 84%, respectively, were erythromycin resistant and clindamycin susceptible. Among erythromycin-resistant S. hominis isolates, 68% of these strains showed the inducible MLSB phenotype. Four isolates harbouring the msr(A) genes alone displayed the MSB phenotype. These studies indicated that resistance to MLSB in S. hominis is mostly based on the ribosomal target modification mechanism mediated by erm genes, mainly the erm(C), and enzymatic drug inactivation mediated by lnu(A).


Subject(s)
Anti-Bacterial Agents/pharmacology , Lincosamides/pharmacology , Macrolides/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus hominis/drug effects , Streptogramins/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Multiple, Bacterial , Humans , Methyltransferases/genetics , Methyltransferases/metabolism , Microbial Sensitivity Tests , Staphylococcus hominis/classification , Staphylococcus hominis/enzymology , Staphylococcus hominis/genetics
7.
PLoS One ; 10(12): e0144684, 2015.
Article in English | MEDLINE | ID: mdl-26659110

ABSTRACT

OBJECTIVES: We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood. METHODS: The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method. RESULTS: Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol. CONCLUSIONS: S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Biofilms/drug effects , Drug Resistance, Bacterial/genetics , Gene Expression Regulation, Bacterial , Staphylococcus hominis/genetics , Amidohydrolases/genetics , Amidohydrolases/metabolism , Bacteremia/drug therapy , Bacteremia/microbiology , Bacterial Proteins/metabolism , Biofilms/growth & development , Cefoxitin/pharmacology , Chromosomes, Bacterial/chemistry , Chromosomes, Bacterial/metabolism , Ciprofloxacin/pharmacology , Humans , Methicillin/pharmacology , Microbial Sensitivity Tests , Operon , Oxacillin/pharmacology , Penicillin-Binding Proteins/genetics , Penicillin-Binding Proteins/metabolism , Phylogeny , Plankton/drug effects , Plankton/growth & development , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/isolation & purification , Vancomycin/pharmacology
8.
Indian J Pathol Microbiol ; 57(2): 275-7, 2014.
Article in English | MEDLINE | ID: mdl-24943764

ABSTRACT

A new subspecies of Staphylococcus hominis described by Kloos et al. in 1998 and named S. hominis subsp. novobiosepticus (SHN) has been implicated in nosocomial outbreaks. Multidrug resistance, including resistance to novobiocin and oxacillin, is a particularly important feature of SHN. In our institute, we encountered 13 cases of S. hominis subsp. hominis in cancer patients with septicemia, of which seven were methicillin resistant. The isolates were identified by VITEK ® 2 compact automated system, using GP REF 21342 identification card and antimicrobial susceptibility testing card P-628. The biochemical reactions and antibiotic susceptibility pattern of the seven methicillin-resistant isolates were re-analyzed and patient details were re-checked to finally identify them as SHN. The increasing number of cases reporting isolation of SHN from biological specimens point to potential virulence and clinical importance of this bacterium.


Subject(s)
Drug Resistance, Multiple, Bacterial , Neoplasms/complications , Sepsis/diagnosis , Sepsis/microbiology , Staphylococcal Infections/diagnosis , Staphylococcal Infections/microbiology , Staphylococcus hominis/isolation & purification , Adolescent , Adult , Aged , Anti-Bacterial Agents/pharmacology , Automation, Laboratory/methods , Bacterial Typing Techniques , Child , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Sepsis/pathology , Staphylococcal Infections/pathology , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects
9.
PLoS One ; 8(6): e66496, 2013.
Article in English | MEDLINE | ID: mdl-23776678

ABSTRACT

Staphylococcus hominis is a commensal resident of human skin and an opportunistic pathogen. The species is subdivided into two subspecies, S. hominis subsp. hominis and S. hominis subsp. novobiosepticus, which are difficult to distinguish. To investigate the evolution and epidemiology of S. hominis, a total of 108 isolates collected from 10 countries over 40 years were characterized by classical phenotypic methods and genetic methods. One nonsynonymous mutation in gyrB, scored with a novel SNP typing assay, had a perfect association with the novobiocin-resistant phenotype. A multilocus sequence typing (MLST) scheme was developed from six housekeeping gene fragments, and revealed relatively high levels of genetic diversity and a significant impact of recombination on S. hominis population structure. Among the 40 sequence types (STs) identified by MLST, three STs (ST2, ST16 and ST23) were S. hominis subsp. novobiosepticus, and they distinguished between isolates from different outbreaks, whereas 37 other STs were S. hominis subsp. hominis, one of which was widely disseminated (ST1). A modified PCR assay was developed to detect the presence of ccrAB4 from the SCCmec genetic element. S. hominis subsp. novobiosepticus isolates were oxacillin-resistant and carriers of specific components of SCCmec (mecA class A, ccrAB3, ccrAB4, ccrC), whereas S. hominis subsp. hominis included both oxacillin-sensitive and -resistant isolates and a more diverse array of SCCmec components. Surprisingly, phylogenetic analyses indicated that S. hominis subsp. novobiosepticus may be a polyphyletic and, hence, artificial taxon. In summary, these results revealed the genetic diversity of S. hominis, the identities of outbreak-causing clones, and the evolutionary relationships between subspecies and clones. The pathogenic lifestyle attributed to S. hominis subsp. novobiosepticus may have originated on more than one occasion.


Subject(s)
Multilocus Sequence Typing/methods , Staphylococcus hominis/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Genetic Variation , Phylogeny , Staphylococcus hominis/classification
10.
PLoS One ; 8(4): e61161, 2013.
Article in English | MEDLINE | ID: mdl-23585877

ABSTRACT

BACKGROUND: Among Coagulase-Negative Staphylococci (CoNS), Staphylococcus hominis represents the third most common organism recoverable from the blood of immunocompromised patients. The aim of this study was to characterize biofilm formation, antibiotic resistance, define the SCCmec (Staphylococcal Chromosomal Cassette mec) type, and genetic relatedness of clinical S. hominis isolates. METHODOLOGY: S. hominis blood isolates (n = 21) were screened for biofilm formation using crystal violet staining. Methicillin resistance was evaluated using the cefoxitin disk test and the mecA gene was detected by PCR. Antibiotic resistance was determined by the broth microdilution method. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE) and SCCmec typed by multiplex PCR using two different methodologies described for Staphylococcus aureus. RESULTS: Of the S. hominis isolates screened, 47.6% (10/21) were categorized as strong biofilm producers and 23.8% (5/21) as weak producers. Furthermore, 81% (17/21) of the isolates were methicillin resistant and mecA gene carriers. Resistance to ampicillin, erythromycin, and trimethoprim was observed in >70% of isolates screened. Each isolate showed a different PFGE macrorestriction pattern with similarity ranging between 0-95%. Among mecA-positive isolates, 14 (82%) harbored a non-typeable SCCmec type: eight isolates were not positive for any ccr complex; four contained the mec complex A ccrAB1 and ccrC, one isolate contained mec complex A, ccrAB4 and ccrC, and one isolate contained the mec complex A, ccrAB1, ccrAB4, and ccrC. Two isolates harbored the association: mec complex A and ccrAB1. Only one strain was typeable as SCCmec III. CONCLUSIONS: The S. hominis isolates analyzed were variable biofilm producers had a high prevalence of methicillin resistance and resistance to other antibiotics, and high genetic diversity. The results of this study strongly suggested that S. hominis isolates harbor new SCCmec structural elements and might be reservoirs of ccrC1 in addition to ccrAB1 and mec complex A.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Bacterial Proteins/metabolism , Bacterial Typing Techniques , Biofilms/drug effects , Biofilms/growth & development , DNA, Bacterial/classification , Genetic Variation , Humans , Methicillin Resistance , Microbial Sensitivity Tests , Phylogeny , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus hominis/classification , Staphylococcus hominis/isolation & purification
11.
J Med Microbiol ; 62(Pt 6): 845-850, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23449871

ABSTRACT

This study reports on the emergence of cfr-harbouring coagulase-negative staphylococci (CoNS) among patients who received linezolid therapy in two hospitals in Hangzhou, China. The mechanisms of resistance and transmission were analysed for these resistant isolates. Eight Staphylococcus capitis isolates, one Staphylococcus epidermidis isolate and one Staphylococcus hominis isolate, obtained from patients who had received linezolid therapy in two hospitals in Hangzhou, China, were confirmed as linezolid resistant, with MICs ranging from 8 to >256 mg l(-1). The linezolid usage data of the ten patients before isolation of the linezolid-resistant CoNS were collected. PFGE analysis showed that the eight S. capitis isolates from the two hospitals belonged to the same clone. Nine of the linezolid-resistant CoNS isolates carried the cfr gene, which was located on plasmids of a similar size. A 5.3 kb fragment containing the cfr gene, revealing 99 % identity to the sequence of the cfr-harbouring plasmid pSS-01 reported previously, was determined by PCR mapping for all cfr-positive isolates, and the cfr gene was flanked by two copies of IS256-like elements. Thus, these results document the emergence of linezolid-resistant CoNS isolates carrying the cfr gene in Hangzhou, China. Effective nosocomial infection control strategies and the judicious use of antibiotics will be required to prevent further spread of this resistance mechanism.


Subject(s)
Acetamides/pharmacology , Anti-Infective Agents/pharmacology , Bacterial Proteins/genetics , Coagulase/metabolism , Drug Resistance, Multiple, Bacterial/genetics , Oxazolidinones/pharmacology , Staphylococcus/drug effects , Aged , Aged, 80 and over , Bacterial Proteins/metabolism , China , Cross Infection/epidemiology , Cross Infection/microbiology , Humans , Linezolid , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Sequence Data , Plasmids/genetics , Sequence Analysis, DNA , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus/classification , Staphylococcus/enzymology , Staphylococcus/isolation & purification , Staphylococcus epidermidis/classification , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/enzymology , Staphylococcus epidermidis/isolation & purification , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/enzymology , Staphylococcus hominis/isolation & purification
12.
Pediatr Infect Dis J ; 32(4): 389-93, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23080292

ABSTRACT

BACKGROUND: Coagulase-negative staphylococci (CoNS) are the first colonizers of gastrointestinal tract (GIT) and the commonest cause of late-onset sepsis (LOS) in preterm neonates. Intravascular catheters are considered a major source of CoNS bacteremia. However, several cases of LOS remain without an identified source. To elucidate whether GIT could be a potential source of invasive strains, we aimed to assess the molecular similarity between CoNS from blood and GIT in preterm neonates with LOS. METHODS: Altogether 22 blood and 53 GIT isolates collected from 22 neonates with LOS caused by CoNS (Staphylococcus haemolyticus in 13, Staphylococcus epidermidis in 7 and Staphylococcus hominis in 2 patients) were included. Rectal swabs were collected twice weekly from birth, but only isolates obtained before LOS were analyzed. S. epidermidis isolates were typed by multilocus variable number of tandem repeats analysis and multilocus sequence typing, S. haemolyticus by pulsed-field gel electrophoresis. RESULTS: Eighteen of 22 neonates had the same CoNS species in GIT and bloodstream; all these isolates from them (altogether 18 blood and 28 GIT isolates) underwent typing. The genotypic similarity between bloodstream and ≥1 antecedent GIT isolates was observed in 13 of 18 patients-3 of 7 with S. epidermidis and 10 of 11 with S. haemolyticus infection. The concordant GIT isolates were collected 0-7 days before the positive blood culture. CONCLUSIONS: The similarity between CoNS from GIT and bloodstream indicates that preterm neonates harbour invasive strains in GIT before LOS. Whether there is a causal relationship between GIT colonization and LOS remains to be elucidated in further studies.


Subject(s)
Gastrointestinal Tract/microbiology , Molecular Typing , Sepsis/microbiology , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/classification , Staphylococcus haemolyticus/classification , Staphylococcus hominis/classification , Age of Onset , Female , Genotype , Humans , Infant, Newborn , Infant, Premature , Male , Prospective Studies , Staphylococcus epidermidis/genetics , Staphylococcus epidermidis/isolation & purification , Staphylococcus haemolyticus/genetics , Staphylococcus haemolyticus/isolation & purification , Staphylococcus hominis/genetics , Staphylococcus hominis/isolation & purification
13.
Folia Microbiol (Praha) ; 56(2): 122-30, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21431912

ABSTRACT

Polymerase chain reaction (PCR) amplification of antibiotic resistance genes as well as staphylococcal cassette chromosome mec (SCCmec) typing and pulsed-field gel electrophoresis (PFGE) of SmaI macrorestriction fragments of genomic DNA were used to characterize 45 methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolates responsible of bacteremia recovered in patients at the Bone Marrow Transplant Centre of Tunisia in 1998-2007. Among the 45 MRCoNS isolates, Staphylococcus epidermidis was the most prevalent species (75.6%) followed by Staphylococcus haemolyticus (22.2%) and Staphylococcus hominis (2.2%). Extended susceptibility profiles were generated for MRCoNS against 16 antimicrobial agents. Out of 45 mecA-positive strains, 43 (95.6%) were phenotypically methicillin-resistant and two (4.4%) were methicillin-susceptible. The msr(A) was the most prevalent gene (13 isolates; 48.1%) among erythromycin-resistant isolates. The erm(C) was found alone in seven (25.9%) or in combination with both erm(A) and erm(B) in two (7.4%) isolates. The aac(6')-Ie-aph(2″)-Ia was the most prevalent gene among aminoglycoside-resistant isolates, detected alone in 14 isolates (33.3%) isolates, in combination with ant(4')-Ia in 18 (42.8%) isolates, in combination with aph(3')-IIIa in four (9.5%) or with both ant(4')-Ia and aph(3')-IIIa in two (4.7%) isolates. The ant(4')-Ia was detected in three (7.1%) isolates and the aph(3')-IIIa in one (2.4%) isolate. Among tetracycline-resistant isolates, six (85.7%) strains harbored the tet(K) gene and one (14.3%) strain carried tet(K) and tet(M) genes. SCCmec types IV (31%) and III (24.5%), the most prevalent types detected, were found to be more resistant to non-ß-lactam antibiotics. A wide diversity of isolates was observed by PFGE among MRCoNS.


Subject(s)
Bacteremia/microbiology , Methicillin Resistance , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/isolation & purification , Staphylococcus haemolyticus/isolation & purification , Staphylococcus hominis/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Coagulase/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Drug Resistance, Bacterial , Electrophoresis, Gel, Pulsed-Field , Genes, Bacterial , Hematologic Neoplasms/complications , Humans , Microbial Sensitivity Tests , Molecular Typing , Polymerase Chain Reaction/methods , Staphylococcus epidermidis/classification , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/genetics , Staphylococcus haemolyticus/classification , Staphylococcus haemolyticus/drug effects , Staphylococcus haemolyticus/genetics , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Tunisia
14.
J Antimicrob Chemother ; 65(11): 2336-40, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20852270

ABSTRACT

BACKGROUND: Despite 10 years of clinical use, linezolid resistance in Staphylococcus aureus and coagulase-negative staphylococci (CoNS) is still a rare phenomenon. This study reports the mechanisms of resistance and strain types seen in clusters of linezolid-resistant CoNS from two different hospitals in Italy during the period 2008-09. METHODS: Genes associated with linezolid resistance were subjected to molecular analysis and isolates were characterized by PFGE macrorestriction analysis using SmaI. RESULTS: Thirty-three linezolid-resistant isolates of methicillin-resistant CoNS comprising Staphylococcus epidermidis (24), Staphylococcus hominis (5) and Staphylococcus simulans (4) were studied. The isolates showed varying levels of linezolid resistance. Almost all isolates for which linezolid MICs were 64 mg/L possessed point mutations in domain V of 23S rRNA, while isolates for which the MICs were 256 mg/L expressed methylase activity at position A2503 mediated by the cfr gene. Overall, the isolates showed reduced susceptibility to vancomycin (MICs 1-2 mg/L) and 11 of the 33 isolates showed no susceptibility to teicoplanin. These strains were also resistant to chloramphenicol (28 of 33), lincomycin (24 of 33), erythromycin (17 of 33) and quinupristin/dalfopristin (13 of 33). S. epidermidis isolates, showing mutations or methylase modifications, belonged to different PFGE profiles and to two different sequence types (ST2 and ST23), in which the cfr gene was carried on a plasmid of ∼50 kb. CONCLUSIONS: Clinical CoNS strains with resistance to linezolid and other second-line antibiotics, as well as reduced susceptibility to glycopeptides, have emerged in Italy.


Subject(s)
Acetamides/pharmacology , Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial , Oxazolidinones/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus epidermidis/drug effects , Staphylococcus hominis/drug effects , Bacterial Typing Techniques , DNA Fingerprinting , DNA Methylation , DNA Modification Methylases/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Electrophoresis, Gel, Pulsed-Field , Hospitals , Humans , Italy , Linezolid , Microbial Sensitivity Tests , Point Mutation , RNA, Ribosomal, 23S/genetics , Staphylococcus aureus/classification , Staphylococcus aureus/enzymology , Staphylococcus aureus/genetics , Staphylococcus epidermidis/classification , Staphylococcus epidermidis/enzymology , Staphylococcus epidermidis/genetics , Staphylococcus hominis/classification , Staphylococcus hominis/enzymology , Staphylococcus hominis/genetics
15.
Eur J Clin Microbiol Infect Dis ; 29(1): 73-80, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19876662

ABSTRACT

Glycopeptides and linezolid are the most widely used antibiotics to treat infections by methicillin-resistant Staphylococcus spp. We report the presence of various isolates of methicillin-resistant S. hominis subsp. hominis with resistance to linezolid and reduced susceptibility to glycopeptides. We studied ten blood culture isolates of S. hominis subsp. hominis from nine patients admitted to our hospital. Etest was used to study susceptibility to antibiotics commonly prescribed against staphylococci. Domain V region of the 23S rRNA gene was amplified and sequenced to detect possible mutations that confer resistance to linezolid. Pulsed-field gel electrophoresis (PFGE) was used for the clonality study of isolates. All isolates were resistant to oxacillin, gentamicin, levofloxacin, cotrimoxazole, and linezolid, and susceptible to tigecycline and daptomycin. Nine of the isolates were resistant to erythromycin and clindamycin, and showed heterogeneous resistance to glycopeptides. C2190T, G2603T, and G2474T mutations were detected in domain V of the 23S rRNA gene. PFGE showed the presence of two different clones. This report alerts to the possible appearance of clinical strains of methicillin-resistant staphylococci with intermediate resistance to glycopeptides, resistance to linezolid, and multiple resistance to other second-line antibiotics.


Subject(s)
Acetamides/pharmacology , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Glycopeptides/pharmacology , Mutation, Missense , Oxazolidinones/pharmacology , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Bacteremia/microbiology , Bacterial Typing Techniques , Blood/microbiology , Cross Infection/microbiology , DNA Mutational Analysis , Electrophoresis, Gel, Pulsed-Field , Genotype , Humans , Intensive Care Units , Linezolid , Microbial Sensitivity Tests , RNA, Bacterial/genetics , RNA, Ribosomal, 23S/genetics , Sequence Analysis, DNA , Staphylococcal Infections/microbiology , Staphylococcus hominis/classification , Staphylococcus hominis/isolation & purification
16.
J Antimicrob Chemother ; 62(6): 1222-6, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18775890

ABSTRACT

OBJECTIVES: To report the isolation of six Staphylococcus hominis subsp. novobiosepticus (SHN) strains from hospitalized patients with bloodstream infections in two Brazilian hospitals and to characterize their susceptibility profile to several antimicrobials. METHODS: Species identification was performed by biochemical methods and sodA gene sequencing. The MICs of antimicrobials were determined by broth and agar dilution methods and by Etest. Isolates were typed by PFGE and PCR amplification was used to detect the ccr gene complex and the mec class. Morphometric evaluation of cell wall was performed by transmission electron microscopy (TEM). RESULTS: Susceptibility profiles indicated that the majority of isolates (five) were multidrug-resistant. Overlapping and multiplex PCR showed that five out of the six strains harboured SCCmec type III with class A mec and type 3 ccr. The initial vancomycin MIC value of 4 mg/L for these strains increased to 16-32 mg/L after growth for 10 days in BHI broth supplemented with this antimicrobial. TEM indicated that vancomycin resistance was associated with cell wall thickening and to another mechanism not fully elucidated. Only one SHN strain was oxacillin- and vancomycin-susceptible. The nosocomial infections in at least five of the patients from both hospitals were caused by a single clone of SHN. CONCLUSIONS: It is very important to consider SHN strains as the cause of nosocomial infections. The clinical implications resulting from the pattern of multidrug resistance in these strains may be complicated by the emergence of vancomycin resistance.


Subject(s)
Bacteremia/microbiology , Cross Infection/microbiology , Staphylococcal Infections/microbiology , Staphylococcus hominis/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacteremia/epidemiology , Bacterial Proteins/genetics , Bacterial Typing Techniques , Brazil/epidemiology , Cell Wall/ultrastructure , Cluster Analysis , Cross Infection/epidemiology , DNA Fingerprinting , Drug Resistance, Multiple, Bacterial , Electrophoresis, Gel, Pulsed-Field , Genotype , Hospitals , Humans , Microbial Sensitivity Tests , Microscopy, Electron, Transmission , Polymerase Chain Reaction , Sequence Analysis, DNA , Staphylococcal Infections/epidemiology , Staphylococcus hominis/classification , Staphylococcus hominis/drug effects , Staphylococcus hominis/genetics , Superoxide Dismutase/genetics
18.
J Clin Microbiol ; 43(9): 4877-9, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16145165

ABSTRACT

From 2002 to 2003, 32 isolates of Staphylococcus hominis subsp. novobiosepticus (SHN) were recovered from 21 patients, 18 of whom were neonates, with 13 considered to have late-onset SHN sepsis. All isolates from neonates had an indistinguishable pulsed-field gel electrophoresis pattern. Our data support SHN as an important nosocomial pathogen in neonates.


Subject(s)
Bacteremia/transmission , Cross Infection/transmission , Infant, Premature, Diseases/microbiology , Intensive Care Units, Neonatal , Staphylococcal Infections/transmission , Staphylococcus hominis/isolation & purification , Adolescent , Adult , Bacteremia/microbiology , Cross Infection/microbiology , Female , Humans , Infant, Newborn , Infant, Premature , Male , Staphylococcal Infections/microbiology , Staphylococcus hominis/classification , Staphylococcus hominis/genetics
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