Propolis is a natural resinous substance collected by bees from tree exudates and secretions. Its antimicrobial activity has been investigated and inhibitory action on Staphylococcus aureus growth was evaluated. The in vitro synergism between ethanolic extract of propolis (EEP) and antimicrobial drugs by two susceptibility tests (Kirby and Bauer and E-Test) on 25 S. aureus strains was evaluated. Petri dishes with sub-inhibitory concentrations of EEP were incubated with 13 drugs using Kirby and Bauer method and synergism between EEP and five drugs [choramphenicol (CLO), gentamicin (GEN), netilmicin (NET), tetracycline (TET), and vancomycin (VAN)] was observed. Nine drugs were assayed by the E-test method and five of them exhibited a synergism [CLO, GEN, NET, TET, and clindamycin (CLI)
Although opportunistic pathogens, coagulase-negative staphylococci (CoNS), including Staphylococcus epidermidis and Staphylococcus haemolyticus, have long been regarded as avirulent organisms. The role of toxins in the development of infections caused by CoNS is still controversial. The objective of this study was to characterize the presence of enterotoxin and cytotoxin genes in S. epidermidis and S. haemolyticus isolates obtained from blood cultures. Cytotoxin genes were detected by PCR using novel species-specific primers. Among the 85 S. epidermidis and 84 S. haemolyticus isolates, 95.3% and 79.8%, respectively, carried at least one enterotoxin gene. The most frequent enterotoxin genes were sea (53.3%), seg (64.5%) and sei (67.5%). The seg gene was positively associated with S. epidermidis (p = 0.02), and this species was more toxigenic than S. haemolyticus. The hla/yidD gene was detected in 92.9% of S. epidermidis and the hla gene in 91.7% of S. haemolyticus isolates; hlb was detected in 92.9% of the S. epidermidis isolates and hld in 95.3%. Nosocomial Staphylococcus epidermidis and S. haemolyticus isolates exhibited a high toxigenic potential, mainly containing the non-classical enterotoxin genes seg and sei. The previously unreported detection of hla/yidD and hlb in S. epidermidis and S. haemolyticus using species-specific primers showed that these hemolysin genes differ between CoNS species and that they are highly frequent in blood culture isolates.
Staphylococcal food poisoning is caused by ingestion of enterotoxins preformed in the food contaminated essentially through human manipulation or raw material obtained from animals. Although coagulase-positive Staphylococcus aureus is the main agent responsible for food intoxication, some researches emphasise that coagulase-negative staphylococci (CNS) are able to produce staphylococcal enterotoxins and may be a potential cause of food poisoning. In the present study CNS were isolated from foods and the toxigenic capacity of the strains determined. A total of 88 food samples were analysed and 22.7% were positive for CNS strains. Staphylococcal counts ranged from 3.0 x 10 2 to 1.4 x 10 6 CFU/g or mL of food examined. S. epidermidis predominated among the isolates (40%). Further isolates included S. xylosus (20%), S. warneri (20%), S. saccharolyticus (15%), and S. hominis (5%). Four isolates were positive for enterotoxin genes, as detected by polymerase chain reaction, with sea being the predominant gene. Although no enterotoxin production was detected by the reverse passive latex agglutination method, the data showed that the toxigenic capacity of CNS should not be ignored, requiring investigation of this group of microorganisms in food.
Peritonitis caused by Staphylococcus aureus is a serious complication of peritoneal dialysis (PD), which is associated with poor outcome and high PD failure rates. We reviewed the records of 62 S. aureus peritonitis episodes that occurred between 1996 and 2010 in the dialysis unit of a single university hospital and evaluated the host and bacterial factors influencing peritonitis outcome. Peritonitis incidence was calculated for three subsequent 5-year periods and compared using a Poisson regression model. The production of biofilm, enzymes, and toxins was evaluated. Oxacillin resistance was evaluated based on minimum inhibitory concentration and presence of the mecA gene. Logistic regression was used for the analysis of demographic, clinical, and microbiological factors influencing peritonitis outcome. Resolution and death rates were compared with 117 contemporary coagulase-negative staphylococcus (CoNS) episodes. The incidence of S. aureus peritonitis declined significantly over time from 0.13 in 1996–2000 to 0.04 episodes/patient/year in 2006–2010 (p = 0.03). The oxacillin resistance rate was 11.3%. Toxin and enzyme production was expressive, except for enterotoxin D. Biofilm production was positive in 88.7% of strains. The presence of the mecA gene was associated with a higher frequency of fever and abdominal pain. The logistic regression model showed that diabetes mellitus (p = 0.009) and β-hemolysin production (p = 0.006) were independent predictors of non-resolution of infection. The probability of resolution was higher among patients aged 41 to 60 years than among those >60 years (p = 0.02). A trend to higher death rate was observed for S. aureus episodes (9.7%) compared to CoNS episodes (2.5%), (p = 0.08), whereas resolution rates were similar. Despite the decline in incidence, S. aureus peritonitis remains a serious complication of PD that is associated with a high death rate. The outcome of this infection is negatively influenced by host factors such as age and diabetes mellitus. In addition, β-hemolysin production is predictive of non-resolution of infection, suggesting a pathogenic role of this factor in PD-related S. aureus peritonitis.
Background and objectives Coagulase-negative Staphylococcus (CNS) is the most frequent cause of peritoneal dialysis (PD)-related peritonitis in many centers. This study aimed to describe clinical and microbiologic characteristics of 115 CNS episodes and to determine factors influencing the outcome.Design, setting, participants, & measurements This study reviewed the records of 115 CNS peritonitis episodes that occurred in 74 patients between 1994 and 2011 at a single university center. Peritonitis incidences were calculated for three consecutive 6-year periods (P1, 1994-1999; P2, 2000-2005; P3, 2006-2011) and annually. The production of biofilms, enzymes, and toxins was evaluated. Oxacillin resistance was evaluated based on its minimum inhibitory concentration and the presence of the mecA gene.Results The overall incidence of CNS peritonitis was 0.15 episodes per patient per year and did not vary over time (0.12, 0.14, and 0.16 for P1, P2, and P3, respectively; P=0.21). The oxacillin resistance rate was 69.6%. Toxin and enzyme production was infrequent and 36.5% of CNS strains presented the gene encoding biofilm production. The presence of icaAD genes associated with biofilm production was predictive of relapses or repeat episodes (odds ratio [OR], 2.82; 95% confidence interval [95% CI], 1.11 to 7.19; P=0.03). Overall, 70 episodes (60.9%) resolved; oxacillin susceptibility (OR, 4.41; 95% CI, 1.48 to 13.17; P=0.01) and vancomycin use as the first treatment (OR, 22.27; 95% CI, 6.16 to 80.53; P,0.001) were the only independent predictors of resolution.Conclusions Oxacillin resistance and vancomycin use as the first treatment strongly influence the resolution rate in CNS peritonitis, which reinforces the validity of the International Society for Peritoneal Dialysis guidelines on monitoring bacterial resistance to define protocols for initial treatment. These results also suggest that the presence of biofilm is a potential cause of repeat peritonitis episodes.
This study aimed to correlate the presence of ica genes, biofilm formation and antimicrobial resistance in 107 strains of Staphylococcus epidermidis isolated from blood cultures. The isolates were analysed to determine their methicillin resistance, staphylococcal cassette chromosome mec (SCCmec) type, ica genes and biofilm formation and the vancomycin minimum inhibitory concentration (MIC) was measured for isolates and subpopulations growing on vancomycin screen agar. The mecA gene was detected in 81.3% of the S. epidermidis isolated and 48.2% carried SCCmec type III. The complete icaADBC operon was observed in 38.3% of the isolates; of these, 58.5% produced a biofilm. Furthermore, 47.7% of the isolates grew on vancomycin screen agar, with an increase in the MIC in 75.9% of the isolates. Determination of the MIC of subpopulations revealed that 64.7% had an MIC ≥ 4 μg mL-1, including 15.7% with an MIC of 8 μg mL-1 and 2% with an MIC of 16 μg mL-1. The presence of the icaADBC operon, biofilm production and reduced susceptibility to vancomycin were associated with methicillin resistance. This study reveals a high level of methicillin resistance, biofilm formation and reduced susceptibility to vancomycin in subpopulations of S. epidermidis. These findings may explain the selection of multidrug-resistant isolates in hospital settings and the consequent failure of antimicrobial treatment.
Coagulase-negative staphylococci (CNS) DNAse (15.4%), thermonuclease (7.7%), and enterotoxin A, B or C (37.6%). Taking into consideration that the etiological importance of CNS has often been neglected, the present investigation confirmed that these microorganisms should not be ignored or classified as mere contaminants.
Recent increases in the incidence and severity of staphylococcal infections renewed interest in studies that assess the burden of asymptomatic carriage of Staphylococcus aureus in the community setting. We conducted a population-based survey in the city of Botucatu, Brazil (122,000 inhabitants), in order to identify the prevalence of nasal carriage of Staphylococcus aureus (including methicillin-resistant strains). Nasal swabs were obtained from 686 persons over one year of age. Resistance to methicillin was assessed through phenotypic methods, identification of the mecA gene and typing of the Staphylococcal Chromosome Cassette mec (SCCmec). Methicillin-resistant S. aureus (MRSA) isolates were characterized using Pulsed-Field Gel Electrophoresis (PFGE), Multilocus Sequence Typing (MLST) and spa typing. Polymerase chain reaction was applied to identify genes coding for Panton-Valentine Leukocidin (PVL) in isolates. The prevalence of overall S. aureus carriage was 32.7% (95%CI, 29.2%–36.2%). Carriers were significantly younger (mean age, 28.1 versus 36.3 for non-carriers; OR for age, 0.98; 95%CI, 0.97–0.99) and likely to report recent skin infection (OR, 1.85; 95%CI, 1.03–3.34). Carriage of methicillin-resistant S. aureus (MRSA) was found in 0.9% of study subjects (95%CI, 0.4%–1.8%). All MRSA isolates harbored SCCmec type IV, and belonged to spa types t002 or t021, but none among them harbored genes coding for PLV. In MLST, most isolates belonged to clones ST5 or ST1776. However, we found one subject who carried a novel clone, ST2594. Two out of six MRSA carriers had household contacts colonized with isolates similar to theirs. Our study pointed to dissemination of community-associated MRSA among the Brazilian population.
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