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1.
Vet Microbiol ; 259: 109160, 2021 Aug.
Article En | MEDLINE | ID: mdl-34197979

High-level mupirocin resistance (HLMR) is determined by the plasmid-located ileS2 gene flanked by two copies of the insertion sequence 257 (IS257). The molecular epidemiology of high-level mupirocin-resistant isolates could be assessed by the determination of their IS257-ileS2 spacer regions conformation. In this study, 188 isolates of methicillin-resistant staphylococci were subjected to the detection of HLMR, and analysis of the conformation of the IS257-ileS2 spacer regions. Mupirocin resistance was detected in five (2,6%) isolates, among which two were recognized as Staphylococcus pseudintermedius, two as Staphylococcus haemolyticus, and one as Staphylococcus aureus. High-level mupirocin resistance was revealed by the agar disk diffusion method, and MIC values, and was confirmed by the detection of the ileS2 gene. The conformations of the IS257-ileS2 spacer regions were homologous in two S. haemolyticus strains tested. The remaining three isolates showed diverse IS257-ileS2 conformations. The results of this study indicate that HLMR occasionally occurs in staphylococci isolated from companion animals. The heterogeneity and the homogeneity of the IS257-ileS2 spacer regions confirm that the ileS2 gene spread among staphylococci of animal origin by the transfer of different as well as the same plasmids. Surveillance of the occurrence of mupirocin resistance and molecular characterization of resistant isolates are strongly recommended due to the possibility of plasmid-located resistance gene transfer between staphylococci.


Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/genetics , Mupirocin/pharmacology , Pets/microbiology , Staphylococcal Infections/veterinary , Animals , Cats/microbiology , Coagulase/biosynthesis , DNA Transposable Elements , Dogs/microbiology , Genes, Bacterial , Methicillin-Resistant Staphylococcus aureus/enzymology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Microbial Sensitivity Tests , Plasmids/genetics , Staphylococcal Infections/microbiology , Staphylococcus/classification , Staphylococcus/drug effects , Staphylococcus/genetics
2.
Microb Drug Resist ; 26(5): 520-530, 2020 May.
Article En | MEDLINE | ID: mdl-31750778

Staphylococcal food poisoning is considered to be one of the most common foodborne illnesses worldwide. Because milk is rich in nutrients and its neutral pH, it leads to the growth of various bacteria. To date, the correlation between enterotoxigenic potential in Staphylococcus species and antimicrobial resistance (AMR), using bioinformatics analysis in buffalo and cow raw milk and the possible health risks from these bacteria, has not been examined in Egypt. A total of 42 Staphylococcus isolates representing 12 coagulase-positive staphylococci (Staphylococcus aureus and Staphylococcus intermedius) and 30 coagulase-negative staphylococci (Staphylococcus capitis, Staphylococcus xylosus, Staphylococcus carnosus, Staphylococcus saccharolyticus, and Staphylococcus auricularis) were isolated. An assay of the antimicrobial resistance phenotypes indicated low resistance against vancomycin (9.5%). The blaZ gene was associated with penicillin G and methicillin resistance and not with sulbactam + ampicillin. The presence of the gene ermB presented the correlation with erythromycin resistance and tetK with tetracycline resistance (correlation index: 0.57 and 0.49, respectively), despite the absence of the same behavior for ermC and tetM, respectively. Interestingly, the gene mecA was not correlated with resistance to methicillin or any other ß-lactam. Correlation showed that slime-producing isolates had more resistance to antibiotics than those of nonslime producers. The multiple correlations between antibiotic resistance phenotypes and resistance genes indicate a complex nature of resistance in Staphylococcus species. The antimicrobial resistance could potentially spread to the community and thus, the resistance of Staphylococcus species to various antibiotics does not depend only on the use of a single antimicrobial, but also extends to other unrelated classes of antimicrobials.


Anti-Bacterial Agents/pharmacology , Coagulase/biosynthesis , Drug Resistance, Bacterial/genetics , Milk/microbiology , Staphylococcus/genetics , beta-Lactamases/genetics , Animals , Buffaloes , Cattle , Computational Biology , Drug Resistance, Bacterial/drug effects , Egypt , Hemolysin Proteins/biosynthesis , Microbial Sensitivity Tests , Staphylococcus/drug effects , Staphylococcus/isolation & purification
3.
Vet Microbiol ; 219: 17-22, 2018 Jun.
Article En | MEDLINE | ID: mdl-29778192

Methicillin-resistant coagulase-negative staphylococci (MRCoNS) have recently emerged as a significant cause of bovine mastitis worldwide. Here we describe the isolation of MRCoNS from cases of bovine mastitis from a single dairy farm in Australia. Fourteen CoNS isolates were identified as MRCoNS on the basis of having an oxacillin MIC of ≥0.5 µg/mL. The isolates were speciated as S. chromogenes (n = 1) S. fleurettii (n = 1), S. haemolyticus (n = 2), S. sciuri (n = 5), S. simulans (n = 1) S. succinus (n = 2) and S. xylosus (n = 2). Five of the isolates (S. fleuretti, S. haemolyticus S. sciuri and two S. succinus) were mecA-positive. We also detected a previously described S. sciuri mecA homolog in four oxacillin-resistant S. sciuri isolates. The remainder of the putative MRCoNS did not contain any mecA-related resistance determinants in their genomes. Comparative genomic analysis of three previously published S. sciuri isolates, from humans, a squirrel and a cereal crop (rice), and a representative isolate from our study demonstrated clustering and a high degree of genetic homogeneity (>95%), suggesting S. sciuri has low host specificity. In conclusion, CoNS, in particular S. sciuri, may act as a reservoir for SCCmec elements that can easily be spread between different host species by direct cross-infection.


Genome, Bacterial/genetics , Mastitis, Bovine/microbiology , Methicillin Resistance/genetics , Methicillin/pharmacology , Staphylococcal Infections/veterinary , Staphylococcus/genetics , Animals , Anti-Bacterial Agents/pharmacology , Australia/epidemiology , Cattle , Coagulase/biosynthesis , Coagulase/deficiency , DNA, Bacterial/genetics , Farms , Female , Microbial Sensitivity Tests , Penicillin-Binding Proteins/genetics , Staphylococcal Infections/epidemiology , Staphylococcal Infections/transmission , Staphylococcal Infections/virology , Staphylococcus/classification , Staphylococcus/drug effects , Staphylococcus/enzymology , Whole Genome Sequencing
4.
Vaccine ; 35(8): 1132-1139, 2017 02 22.
Article En | MEDLINE | ID: mdl-28143674

BACKGROUND: Staphylococcus aureus is a leading cause of healthcare-associated infections. No preventive vaccine is currently licensed. SA4Ag is an investigational 4-antigen S. aureus vaccine, composed of capsular polysaccharide conjugates of serotypes 5 and 8 (CP5 and CP8), recombinant surface protein clumping factor A (rmClfA), and recombinant manganese transporter protein C (rMntC). This Phase 1 study aimed to confirm the safety and immunogenicity of SA4Ag produced by the final manufacturing process before efficacy study initiation in a surgical population. METHODS: Healthy adults (18-<65years) received one intramuscular SA4Ag injection. Serum functional antibodies were measured at baseline and Day 29 post-vaccination. An opsonophagocytic activity (OPA) assay measured the ability of vaccine-induced antibodies to CP5 and CP8 to kill S. aureus clinical isolates. For MntC and ClfA, antigen-specific immunogenicity was assessed via competitive Luminex® immunoassay (cLIA) and via fibrinogen-binding inhibition (FBI) assay for ClfA only. Reactogenicity and adverse event data were collected. RESULTS: One hundred participants were vaccinated. SA4Ag was well tolerated, with a satisfactory safety profile. On Day 29, OPA geometric mean titers (GMTs) were 45,738 (CP5, 95% CI: 38,078-54,940) and 42,652 (CP8, 95% CI: 32,792-55,477), consistent with 69.2- and 28.9-fold rises in bacteria-killing antibodies, respectively; cLIA GMTs were 2064.4 (MntC, 95% CI: 1518.2-2807.0) and 3081.4 (ClfA, 95% CI: 2422.2-3920.0), consistent with 19.6- and 12.3-fold rises, respectively. Similar to cLIA results, ClfA FBI titers rose 11.0-fold (GMT: 672.2, 95% CI: 499.8-904.2). The vast majority of participants achieved the pre-defined biologically relevant thresholds: CP5: 100%; CP8: 97.9%, ClfA: 87.8%; and MntC 96.9%. CONCLUSIONS: SA4Ag was safe, well tolerated, and rapidly induced high levels of bacteria-killing antibodies in healthy adults. A Phase 2B efficacy trial in adults (18-85years) undergoing elective spinal fusion is ongoing to assess SA4Ag's ability to prevent postoperative invasive surgical site and bloodstream infections caused by S. aureus. Clinicaltrials.gov Identifier: NCT02364596.


Antibodies, Bacterial/blood , Antigens, Bacterial/administration & dosage , Staphylococcal Vaccines/administration & dosage , Staphylococcus aureus/immunology , Vaccination , Adolescent , Adult , Aged , Antigens, Bacterial/chemistry , Antigens, Bacterial/immunology , Coagulase/administration & dosage , Coagulase/biosynthesis , Coagulase/genetics , Female , Healthy Volunteers , Humans , Immunogenicity, Vaccine , Injections, Intramuscular , Male , Middle Aged , Patient Safety , Periplasmic Binding Proteins/administration & dosage , Periplasmic Binding Proteins/biosynthesis , Periplasmic Binding Proteins/genetics , Polysaccharides, Bacterial/administration & dosage , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/immunology , Recombinant Proteins/administration & dosage , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Serogroup , Staphylococcal Infections/prevention & control , Staphylococcal Vaccines/biosynthesis , Staphylococcal Vaccines/genetics , Staphylococcus aureus/chemistry , Vaccines, Conjugate
5.
Rev Soc Bras Med Trop ; 49(3): 292-9, 2016.
Article En | MEDLINE | ID: mdl-27384825

INTRODUCTION: Coagulase-negative staphylococci (CoNS) are the most prevalent pathogens in nosocomial infections and may serve as a reservoir of mobile genetic elements such as the staphylococcal cassette chromosome mec (SCCmec) encoding methicillin resistance. Molecular characterization of SCCmec types combined with advanced molecular typing techniques may provide essential information for understanding the evolution and epidemiology of CoNS infections. We therefore aimed to investigate the SCCmec distribution, multidrug-resistance (MDR), and biofilm formation in CoNS blood culture isolates from a hospital in Southern Brazil. METHODS: We analyzed 136 CoNS blood culture isolates obtained during 2002-2004 from patients admitted to a tertiary care hospital in Brazil. SCCmec types I to V were determined using multiplex PCR. The clonal relationship of Staphylococcus epidermidis was determined using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Molecular epidemiological data were interpreted along with data on biofilm formation, presence of the icaD gene, and MDR. RESULTS: The most prevalent species were S. epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis harboring mainly SCCmec types II, III, and V. Overall, the presence of multiple SCCmec was associated with non-MDR, except for S. epidermidis. S. epidermidis isolates showed a high prevalence of icaD, but had low phenotypic biofilm formation. PFGE and MLST revealed high genetic diversity in the S. epidermidis population. CONCLUSIONS: Our results suggest a major shift in SCCmec types within a short period and reveal a different behavior of S. epidermidis with regard to the association between the presence of multiple SCCmec types and MDR profile.


Chromosomes, Bacterial/genetics , DNA, Bacterial/genetics , Genetic Variation/genetics , Staphylococcus/classification , Biofilms/growth & development , Coagulase/biosynthesis , Electrophoresis, Gel, Pulsed-Field , Humans , Multilocus Sequence Typing , Staphylococcus/enzymology , Staphylococcus/genetics
7.
Rev. Soc. Bras. Med. Trop ; 49(3): 292-299, tab, graf
Article En | LILACS | ID: lil-785791

Abstract: INTRODUCTION: Coagulase-negative staphylococci (CoNS) are the most prevalent pathogens in nosocomial infections and may serve as a reservoir of mobile genetic elements such as the staphylococcal cassette chromosome mec (SCCmec) encoding methicillin resistance. Molecular characterization of SCCmec types combined with advanced molecular typing techniques may provide essential information for understanding the evolution and epidemiology of CoNS infections. We therefore aimed to investigate the SCCmec distribution, multidrug-resistance (MDR), and biofilm formation in CoNS blood culture isolates from a hospital in Southern Brazil. METHODS: We analyzed 136 CoNS blood culture isolates obtained during 2002-2004 from patients admitted to a tertiary care hospital in Brazil. SCCmec types I to V were determined using multiplex PCR. The clonal relationship of Staphylococcus epidermidis was determined using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Molecular epidemiological data were interpreted along with data on biofilm formation, presence of the icaD gene, and MDR. RESULTS: The most prevalent species were S. epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis harboring mainly SCCmec types II, III, and V. Overall, the presence of multiple SCCmec was associated with non-MDR, except for S. epidermidis. S. epidermidis isolates showed a high prevalence of icaD, but had low phenotypic biofilm formation. PFGE and MLST revealed high genetic diversity in the S. epidermidis population. CONCLUSIONS: Our results suggest a major shift in SCCmec types within a short period and reveal a different behavior of S. epidermidis with regard to the association between the presence of multiple SCCmec types and MDR profile.


Humans , Staphylococcus/classification , Genetic Variation/genetics , DNA, Bacterial/genetics , Chromosomes, Bacterial/genetics , Staphylococcus/enzymology , Staphylococcus/genetics , Electrophoresis, Gel, Pulsed-Field , Coagulase/biosynthesis , Biofilms/growth & development , Multilocus Sequence Typing
8.
APMIS ; 123(10): 867-71, 2015 Oct.
Article En | MEDLINE | ID: mdl-26227107

The aim of this article were to determinate the mechanism of linezolid resistance in coagulase-negative methicillin-resistant staphylococci from hospitals in the northeast of Brazil. We identified the isolates using VITEK(®) 2 and MALDI-TOF. Susceptibility to antibiotics was measured by the disk-diffusion method and by Etest(®) . Extraction of the whole genome DNA was performed, followed by screening of all the strains for the presence of mecA and cfr genes. The domain V region of 23S rRNA gene was sequenced and then aligned with a linezolid-susceptible reference strain. Pulsed-field gel electrophoresis (PFGE) macro-restriction analysis was performed. Three linezolid-resistant Staphylococcus hominis and two linezolid-resistant Staphylococcus epidermidis strains were analyzed. The isolates showed two point mutations in the V region of the 23S rRNA gene (C2190T and G2603T). We did not detect the cfr gene in any isolate by PCR. The S. hominis showed the same pulsotype, while the S. epidermidis did not present any genetic relation to each other. In conclusion, this study revealed three S. hominis and two S. epidermidis strains with resistance to linezolid due to a double mutation (C2190T and G2603T) in the domain V of the 23S rRNA gene. For the first time, the mutation of C2190T in S. epidermidis is described. This study also revealed the clonal spread of a S. hominis pulsotype between three public hospitals in the city of Natal, Brazil. These findings highlight the importance of continued vigilance of linezolid resistance in staphylococci.


Linezolid/pharmacology , Methicillin/pharmacology , RNA, Ribosomal, 23S/genetics , Staphylococcus epidermidis/drug effects , Staphylococcus hominis/drug effects , Anti-Bacterial Agents , Base Sequence , Coagulase/biosynthesis , Humans , Methicillin Resistance/genetics , Microbial Sensitivity Tests , Mutation/genetics , Sequence Analysis, RNA , Staphylococcus epidermidis/genetics , Staphylococcus epidermidis/isolation & purification , Staphylococcus hominis/genetics , Staphylococcus hominis/isolation & purification
9.
Mem Inst Oswaldo Cruz ; 108(7): 939-42, 2013 Nov.
Article En | MEDLINE | ID: mdl-24141968

Coagulase-negative staphylococci (CoNS) are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec) type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.


Anti-Bacterial Agents/pharmacology , Oxacillin/pharmacology , Penicillin Resistance , Staphylococcus/drug effects , Coagulase/biosynthesis , Coagulase/genetics , Humans , Infant, Newborn , Methicillin Resistance/genetics , Microbial Sensitivity Tests , Polymerase Chain Reaction , Staphylococcus/classification , Staphylococcus/enzymology , Staphylococcus/genetics
10.
APMIS ; 120(10): 786-93, 2012 Oct.
Article En | MEDLINE | ID: mdl-22958286

A novel murine experimental wound infection model was used to assess the efficacy of multi-component immunization against Staphylococcus aureus infection. Necrotic lesions were induced in mice with venom from Bothrops asper and infected with a low inoculum, 1 × 10(2) CFU. The wound infection model therefore more resembles a clinical case of S. aureus infection compared with conventional infection models where far more bacteria are required. Before infection, mice were immunized with four recombinant S.aureus proteins expressed from Escherichia coli: (i) domains 1-3 of Extracellular adherence protein (Eap), (ii) Efb - D (fusion protein combining Extracellular fibrinogen binding protein (Efb) and a fibronectin binding domain (D) of the fibronectin binding protein (FnBP) and (iii) clumping factor A (ClfA). In the immunized group, lower bacterial colonization, undisturbed crust formation and significantly faster wound healing were found compared with the unimmunized control group. Efb and Eap have previously been found to impair wound healing and neutralization of these proteins by antibodies restores a more natural wound healing process. This effect is further also enhanced by the proposed opsonic activity of antibodies against ClfA and FnBP.


Bothrops , Crotalid Venoms/administration & dosage , Staphylococcal Infections/prevention & control , Staphylococcal Vaccines/immunology , Staphylococcus aureus/immunology , Wound Infection/prevention & control , Adhesins, Bacterial/administration & dosage , Adhesins, Bacterial/biosynthesis , Adhesins, Bacterial/immunology , Animals , Antibodies, Bacterial/biosynthesis , Antibodies, Bacterial/immunology , Bacterial Proteins/administration & dosage , Bacterial Proteins/biosynthesis , Bacterial Proteins/immunology , Coagulase/administration & dosage , Coagulase/biosynthesis , Coagulase/immunology , Escherichia coli , Female , Immunization , Mice , Mice, Inbred BALB C , Models, Animal , Necrosis/immunology , Necrosis/pathology , RNA-Binding Proteins/administration & dosage , RNA-Binding Proteins/biosynthesis , RNA-Binding Proteins/immunology , Recombinant Proteins/administration & dosage , Recombinant Proteins/biosynthesis , Recombinant Proteins/immunology , Staphylococcal Infections/immunology , Staphylococcal Infections/pathology , Staphylococcal Vaccines/administration & dosage , Wound Infection/immunology , Wound Infection/pathology
11.
Clin Microbiol Infect ; 18(9): E355-61, 2012 Sep.
Article En | MEDLINE | ID: mdl-22784359

Coagulase-negative staphylococci (CoNS) are frequent contaminants of blood cultures. We aimed to evaluate the systemic inflammatory response syndrome (SIRS) criteria in patients with CoNS bacteraemia for discrimination between true bloodstream infection (BSI) and contamination. Prospective evaluation was carried out of clinical and laboratory parameters in adults with at least one positive blood culture with CoNS at the University Hospital of Basel between 2003 and 2007. Of 3060 positive blood cultures, 654 episodes of CoNS bacteraemia were identified. Of these, 232 (35%) were considered to be true BSI and 422 (65%) were considered to be contamination. Overall, 80% of study participants had at least one SIRS criterion, fever being the most common, and 49% had at least two SIRS criteria. In the multivariate analysis, independent predictors of BSI were fever or hypothermia (OR 2.93, 95% CI 1.91-4.5), tachycardia (OR 2.29, 95% CI 1.50-3.50), tachypnoea (OR 2.4, 95% CI 1.30-4.43), leucocytosis or leucopenia (OR 4.15, 95% CI 2.17-6.36) and the presence of a central venous line (OR 5.38, 95% CI 3.25-8.88). The probability of BSI increased with each additional SIRS criterion, ranging from 42.4% in patients with only one SIRS criterion to 56.7% for those with two criteria, and 72.3% for patients with three SIRS criteria. A positive blood culture with CoNS most likely represents true BSI if the patient has at least three SIRS criteria or two SIRS criteria and a central venous catheter. These simple bedside criteria may guide decision to treat, decreasing the use of glycopeptides.


Bacteremia/blood , Bacteremia/microbiology , Staphylococcus/classification , Systemic Inflammatory Response Syndrome/blood , Systemic Inflammatory Response Syndrome/microbiology , Aged , Algorithms , Bacteremia/diagnosis , Central Venous Catheters , Coagulase/biosynthesis , Diagnosis, Differential , Female , Humans , Male , Middle Aged , Multivariate Analysis , Prospective Studies , Reproducibility of Results , Staphylococcus/enzymology , Staphylococcus/isolation & purification
12.
Biosci Biotechnol Biochem ; 75(6): 1177-80, 2011.
Article En | MEDLINE | ID: mdl-21670514

This study investigates whether there is a predominant Staphylococcus aureus strain in retail foods and healthy human hands, and examines the relationship between pulsed-field gel electrophoresis (PFGE) banding patterns and the S. aureus characteristics of staphylococcal enterotoxin (SE) type, coagulase type, and ß-lactamase activity. Ninety-four strains of S. aureus isolated from retail foods and healthy human hands were analyzed by PFGE. Several strains isolated from the same shop or a chain store showed identical patterns, indicating that the origins of these strains were identical. After excluding these strains showing identical patterns, 54 strains were used for the PFGE analysis. No spread of a particular clone in the environment surrounding the food was apparent. The PFGE analysis of these 54 strains was classified in 6 lineages (L1-L6). There was no relationship between the PFGE banding pattern and coagulase type or SE type. Eleven (84.6%) of the 13 isolates in PFGE banding pattern L5 did not produce ß-lactamase, suggesting that the production of ß-lactamase influenced a specific PFGE banding pattern.


Coagulase/analysis , Electrophoresis, Gel, Pulsed-Field/methods , Enterotoxins/analysis , Food Handling , Staphylococcus aureus , beta-Lactamases/analysis , Bacterial Typing Techniques , Coagulase/biosynthesis , Enterotoxins/biosynthesis , Food Industry , Humans , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/isolation & purification , beta-Lactamases/biosynthesis
13.
Infect Control Hosp Epidemiol ; 32(6): 623-5, 2011 Jun.
Article En | MEDLINE | ID: mdl-21558778

Of all blood cultures positive for coagulase-negative staphylococci collected in 1 year at an academic hospital, 100 were selected randomly for review and designated true positives or contaminated. For the 85 patients whose cultures were determined to be contaminated, chart abstractions revealed substantial unnecessary antibiotic administration, additional laboratory tests and procedures, and hospital readmissions.


Anti-Bacterial Agents/therapeutic use , Bacteremia/diagnosis , Staphylococcal Infections/diagnosis , Staphylococcus , Unnecessary Procedures , Academic Medical Centers , Bacteremia/drug therapy , Bacteremia/microbiology , Blood Specimen Collection , Coagulase/biosynthesis , False Positive Reactions , Female , Humans , Male , Middle Aged , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus/enzymology , Staphylococcus/isolation & purification
14.
Neonatal Netw ; 30(3): 165-74, 2011.
Article En | MEDLINE | ID: mdl-21576051

Coagulase-negative Staphylococcus (CoNS) bloodstream infection is the most common cause of sepsis in the NICU and can lead to significant morbidity and mortality. There is evidence that hand hygiene using an alcohol-based gel and wearing gloves during patient care, management of central and peripheral intravenous lines using the Centers for Disease Control and Prevention (CDC) guidelines, and a closed medication administration system can reduce the incidence to CoNS sepsis in the (NICU). To successfully apply the evidence and decrease the CoNS infection rate, a systematic process is necessary. One approach to process change that significantly reduced the CoNS infection rate in a health care system with two Level III NICUs included using system thinking; working within a multidisciplinary team; using evidence to revise, develop, and implement policies and procedures; developing staff education programs; and monitoring and providing feedback to all staff members.


Intensive Care Units, Neonatal/standards , Staphylococcal Infections/nursing , Staphylococcal Infections/prevention & control , Coagulase/biosynthesis , Humans , Hygiene/standards , Infant, Newborn , Nursing/methods , Pennsylvania , Staphylococcal Infections/drug therapy , Staphylococcal Infections/enzymology
15.
Eur J Clin Microbiol Infect Dis ; 30(6): 785-8, 2011 Jun.
Article En | MEDLINE | ID: mdl-21249410

The Clinical and Laboratory Standards Institute (CLSI) recommends testing coagulase-negative staphylococci (CoNS) strains to determine resistance against oxacillin by testing for mecA, PBP2a, or with cefoxitin disk. However, discrepant results of resistance to oxacillin and susceptibility to cefoxitin were found. In this study, we aimed to investigate the oxacillin resistance and cefoxitin susceptibility of CoNS in Taiwan. Of 9,017 strains collected from 2005 to 2010, 131 (1.5%) of the isolates were oxacillin-resistant and cefoxitin-susceptible. Species identification was carried out using the Vitek 2 system or 16S ribosomal RNA sequencing. Oxacillin minimum inhibitory concentrations (MICs) were examined by the agar dilution method. The presence of mecA and the activity of ß-lactamase were performed by polymerase chain reaction (PCR) and Cefinase disks, respectively. Overall, 33% (43/129) of the strains carried mecA and 43% (37/86) of mecA-negative isolates tested positive for ß-lactamase. The remaining 49 isolates were negative for both mecA and ß-lactamase, and were mainly Staphylococcus cohnii ssp. urealyticus and S. saprophyticus (oxacillin MICs 0.5-2 µg/ml) obtained from bloodstream and urinary tract infections. Our study suggests that incorrect reporting can be found in CoNS using cefoxitin disk alone to determine the susceptibility to oxacillin, and the strains should be further tested for oxacillin MICs and detection of the mecA gene or ß-lactamase activity.


Anti-Bacterial Agents/pharmacology , Cefoxitin/pharmacology , Oxacillin/pharmacology , Staphylococcus/drug effects , beta-Lactam Resistance , Bacterial Typing Techniques , Coagulase/biosynthesis , Genotype , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Staphylococcal Infections/microbiology , Staphylococcus/classification , Staphylococcus/enzymology , Staphylococcus/isolation & purification , Taiwan
16.
Eur J Clin Microbiol Infect Dis ; 29(5): 591-5, 2010 May.
Article En | MEDLINE | ID: mdl-20221891

In recent years, coagulase-negative staphylococci (CoNS) have been increasingly recognised as causative agents of various infections, especially in immunocompromised patients and related to implanted foreign body materials. CoNS, and especially Staphylococcus epidermidis, transform into a stationary growth phase and produce biofilm when involved in a foreign body infection, making them difficult to eradicate with antimicrobials. Rifampicin has the ability to penetrate biofilm, but resistance may develop rapidly. To reduce the emergence of resistance, rifampicin should be combined with additional antimicrobials, of which several different ones have been proposed, including the relatively new class of antimicrobials, oxazolidinones, represented by linezolid. Thirty-seven CoNS isolates from patients with prosthetic joint infection were investigated by synergy testing using Etest. Nine antimicrobial combinations, based on either rifampicin or linezolid, were tested. For 16 (43%) of the isolates, a synergistic (n = 5), additive (n = 14) and/or antagonistic (n = 11) effect were identified. In conclusion, Etest is an objective and easily performed in vitro method for antimicrobial synergy testing. However, each isolate requires testing for the specific combination considered for treatment.


Acetamides/pharmacology , Anti-Infective Agents/pharmacology , Oxazolidinones/pharmacology , Prosthesis-Related Infections/microbiology , Rifampin/pharmacology , Staphylococcus epidermidis/drug effects , Coagulase/biosynthesis , Drug Synergism , Humans , Joint Prosthesis/adverse effects , Linezolid , Microbial Sensitivity Tests , Reproducibility of Results , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/enzymology , Staphylococcus epidermidis/isolation & purification
17.
J Clin Microbiol ; 48(4): 1428-31, 2010 Apr.
Article En | MEDLINE | ID: mdl-20164270

We studied 315 coagulase-negative Staphylococcus strains recovered prospectively during 240 surgical procedures (206 subjects) from proven or suspected device-associated bone and joint infections. Sixteen strains (5.1%) had decreased susceptibility to glycopeptides: 15 (12 S. epidermidis strains, 2 S. capitis strains, and 1 S. haemolyticus strain) to teicoplanin alone (MIC of 16 mg/liter, n = 9; MIC of 32 mg/liter, n = 6) and one (S. epidermidis) to both teicoplanin and vancomycin (MIC, 16 and 8 mg/liter, respectively). Decreased susceptibility to teicoplanin was more prevalent in "infecting" strains (i.e., strains recovered from >/=2 distinct intraoperative samples) than in "contaminants" (i.e., strains not fulfilling this criterion) (8.1% [12/149] versus 2.4% [4/166], respectively [P = 0.022]). One hundred percent (13/13) of S. epidermidis strains with decreased susceptibility to teicoplanin were resistant to methicillin (versus 112/173 [64.7%] for S. epidermidis strains susceptible to teicoplanin; P = 0.021).


Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Prosthesis-Related Infections/microbiology , Staphylococcal Infections/microbiology , Staphylococcus/drug effects , Teicoplanin/pharmacology , Vancomycin/pharmacology , Adult , Aged , Aged, 80 and over , Coagulase/biosynthesis , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Staphylococcus/enzymology , Young Adult
18.
J Med Microbiol ; 59(Pt 3): 323-329, 2010 Mar.
Article En | MEDLINE | ID: mdl-20007762

The objective of this study was to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) elements in meticillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from a tertiary-care hospital in Mexico and to examine the relationship to drug resistance. Fifty selected MR-CoNS isolates collected from catheters (n=15), blood (n=15), bone (n=9), bronchial lavage (n=2) and urine (n=2) and one isolate each from an abscess, cerebrospinal fluid, eye, pleural effusion, synovial fluid, tracheal aspirate and wound secretion were examined. Susceptibility testing was performed by the broth microdilution method. SCCmec types were determined by multiplex PCR and PFGE was carried out as described previously for Staphylococcus aureus. Among the MR-CoNS strains studied, the most frequently isolated species were Staphylococcus epidermidis (n=26) and Staphylococcus haemolyticus (n=13). Staphylococcus cohnii (n=5), Staphylococcus hominis (n=3), Staphylococcus sciuri (n=1), Staphylococcus pasteuri (n=1) and the recently described species Staphylococcus pettenkoferi (n=1) were also identified. The most frequent MR-CoNS genotype identified was SCCmec type IVa in S. epidermidis isolates, which also showed a high diversity in their PFGE patterns. A clone was found that amplified both SCCmec III and V elements in five isolates examined. The single MR S. pettenkoferi isolate harboured SCCmec type IVd and the single MR S. pasteuri isolate harboured SCCmec type I. The carriage of SCCmec type III was associated with resistance or intermediate resistance to meropenem (P <0.05). These results confirm the high prevalence of S. epidermidis SCCmec IVa and the high genetic diversity among MR-CoNS strains. As far as is known, this is the first report describing the newly identified S. pettenkoferi possessing SCCmec IVd and S. pasteuri harbouring SCCmec type I. MR-CoNS harbouring SCCmec type III were found to be more resistant to meropenem.


Coagulase/biosynthesis , Genetic Variation , Methicillin Resistance , Staphylococcal Infections/microbiology , Staphylococcus/drug effects , Staphylococcus/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Chromosomes, Bacterial , DNA Fingerprinting , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Female , Genotype , Hospitals , Humans , Infant , Infant, Newborn , Male , Mexico , Microbial Sensitivity Tests , Middle Aged , Molecular Sequence Data , Phenotype , Polymerase Chain Reaction , Sequence Analysis, DNA , Staphylococcus/classification , Staphylococcus/isolation & purification , Young Adult
19.
Epidemiol Infect ; 138(5): 645-54, 2010 May.
Article En | MEDLINE | ID: mdl-19961645

Coagulase-negative staphylococci (CNS) are increasingly recognized to cause clinically significant infections, with S. epidermidis often cited as the third most common cause of nosocomial sepsis. Among CNS, there is a high prevalence of methicillin resistance associated with staphylococcal cassette chromosome (SCCmec) elements. Although identical SCCmec types can exist in S. aureus and CNS, some novel classes of SCCmec may be unique to CNS. Differences in the accuracy of identification of CNS species and use of non-standardized methods for the detection of methicillin resistance have led to confusing data in the literature. In addition to the review of SCCmec in CNS, in this paper we report a 2-year surveillance of methicillin-resistant CNS in a tertiary-care hospital in Guadalajara, Mexico.


Chromosomes, Bacterial , Coagulase/biosynthesis , Cross Infection/microbiology , DNA, Bacterial/genetics , Methicillin Resistance , Staphylococcal Infections/microbiology , Staphylococcus/genetics , Bacterial Typing Techniques , Cross Infection/epidemiology , DNA Fingerprinting , Electrophoresis, Gel, Pulsed-Field , Hospitals , Humans , Mexico , Molecular Epidemiology , Staphylococcal Infections/epidemiology , Staphylococcus/classification , Staphylococcus/drug effects , Staphylococcus/isolation & purification
20.
BMC Infect Dis ; 9: 212, 2009 Dec 22.
Article En | MEDLINE | ID: mdl-20028509

BACKGROUND: Peritonitis continues to be the most frequent cause of peritoneal dialysis (PD) failure, with an important impact on patient mortality. Gram-positive cocci such as Staphylococcus epidermidis, other coagulase-negative staphylococci (CoNS), and Staphylococcus aureus are the most frequent etiological agents of PD-associated peritonitis worldwide. The objective of the present study was to compare peritonitis caused by S. aureus and CoNS and to evaluate the factors influencing outcome. METHODS: Records of 86 new episodes of staphylococcal peritonitis that occurred between 1996 and 2000 in the Dialysis unit of a single university hospital were studied (35 due to S. aureus, 24 to S. epidermidis and 27 to other CoNS). The production of slime, lipase, lecithinase, nuclease (DNAse), thermonuclease (TNAse), alpha- and beta-hemolysin, enterotoxins (SEA, SEB, SEC, SED) and toxic shock syndrome toxin-1 (TSST-1) was studied in S. aureus and CoNS. Antimicrobial susceptibility was evaluated based on the minimal inhibitory concentration determined by the E-test. Outcome predictors were evaluated by two logistic regression models. RESULTS: The oxacillin susceptibility rate was 85.7% for S. aureus, 41.6% for S. epidermidis, and 51.8% for other CoNS (p = 0.001). Production of toxins and enzymes, except for enterotoxin A and alpha-hemolysin, was associated with S. aureus episodes (p < 0.001), whereas slime production was positive in 23.5% of CoNS and 8.6% of S. aureus strains (p = 0.0047). The first model did not include enzymes and toxins due to their association with S. aureus. The odds of resolution were 9.5 times higher for S. epidermidis than for S. aureus (p = 0.02) episodes, and were similar for S. epidermidis and other CoNS (p = 0.8). The resolution odds were 68 times higher for non-slime producers (p = 0.001) and were not influenced by oxacillin resistance among vancomycin-treated cases (p = 0.89). In the second model, the resolution rate was similar for S. aureus and S. epidermidis (p = 0.70), and slime (p = 0.001) and alpha-hemolysin (p = 0.04) production were independent predictors of non-resolution. CONCLUSION: Bacterial species and virulence factors rather than antibiotic resistance influence the outcome of staphylococcal peritonitis.


Peritoneal Dialysis, Continuous Ambulatory , Peritonitis/etiology , Peritonitis/microbiology , Staphylococcal Infections/complications , Virulence Factors/metabolism , Adult , Coagulase/biosynthesis , Drug Resistance, Multiple, Bacterial , Female , Humans , Logistic Models , Male , Microbial Sensitivity Tests , Middle Aged , Peritonitis/mortality , Staphylococcal Infections/microbiology , Staphylococcus aureus/metabolism , Staphylococcus aureus/pathogenicity , Staphylococcus epidermidis/metabolism , Staphylococcus epidermidis/pathogenicity , Treatment Outcome , Young Adult
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