Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli.

(1) Background: Pathogenic Escherichia coli are divided into two groups: diarrheagenic (DEC) and extraintestinal pathogenic (ExPEC) E. coli. ExPEC causing urinary tract infections (UTIs) are termed uropathogenic E. coli (UPEC) and are the most common cause of UTIs worldwide. (2) Methods: Here, we characterized 112 UPEC in terms of phylogroup, serotype, the presence of virulence factor-encoding genes, and antimicrobial resistance. (3) Results: The majority of the isolates were assigned into the phylogroup B2 (41.07%), and the serogroups O6 (12.5%) and O25 (8.9%) were the most frequent. Five hybrid UPEC (4.5%), with markers from two DEC pathotypes, i.e., atypical enteropathogenic (aEPEC) and enteroaggregative (EAEC) E. coli, were identified, and designated UPEC/aEPEC (one isolate) and UPEC/EAEC (four isolates), respectively. Three UPEC/EAEC harbored genes from the pap operon, and the UPEC/aEPEC carried ibeA. The highest resistance rates were observed for ampicillin (46.4%) and trimethoprim/sulfamethoxazole (34.8%), while 99.1% of the isolates were susceptible to nitrofurantoin and/or fosfomycin. Moreover, 9.8% of the isolates were identified as Extended Spectrum ß-Lactamase producers, including one hybrid UPEC/EAEC. (4) Conclusion: Our data reinforce that hybrid UPEC/DEC are circulating in the city of Botucatu, Brazil, as uropathogens. However, how and whether these combinations of genes influence their pathogenicity is a question that remains to be elucidated.

Evaluation of MALDI-TOF VITEK®MS and VITEK® 2 system for the identification of Staphylococcus saprophyticus.

AIM: To compare two identification methods for coagulase-negative staphylococci (CoNS) isolated from patients with urinary tract infections, VITEK® 2 and MALDI-TOF VITEK®MS, with genotypic identification by internal transcribed spacer PCR (ITS-PCR). RESULTS: A total of 217 CoNS isolates were studied. Agreement of the VITEK® 2 system with ITS-PCR was 84.8%, with 98% sensitivity and 100% specificity. Thirty-one of the 33 strains incorrectly identified by VITEK® 2 belonged to the species Staphylococcus saprophyticus. MALDI-TOF VITEK®MS showed an excellent correlation with ITS-PCR since it correctly identified all CoNS isolates. CONCLUSION: MALDI-TOF VITEK®MS is more accurate than the automated VITEK® 2 system in identifying CoNS isolated from urinary tract infections to species level, particularly urinary isolates of S. saprophyticus.

Diarrheagenic Escherichia coli pathotypes investigation revealed atypical enteropathogenic E. coli as putative emerging diarrheal agents in children living in Botucatu, São Paulo State, Brazil.

The aim of the present study was to investigate the prevalence of Diarrheagenic Escherichia coli (DEC) pathotypes, a leading cause of diarrhea worldwide, among diarrheal and healthy children, up to 5 years of age, living in the city of Botucatu, São Paulo, Brazil. DEC, investigated by PCR detection of virulence factor-encoding genes associated with the distinct pathotypes, was isolated from 18.0% of the patients, and 19.0% of the controls, with enteroaggregative E. coli (EAEC), the most frequent pathotype, being detected in equal proportion between patients and controls (10.0%). Among the enteropathogenic E. coli (EPEC) isolates, only one isolate was able to produce the localized adherence pattern to HeLa cells, being thus the only typical EPEC identified. All the remaining EPEC were classified as atypical (aEPEC), and detected in 8.0% and 8.5% of the patients and controls, respectively. Regarding the serotypes, 26.5% of the analyzed EPEC isolates belonged to classical EPEC-serogroups, and the only two STEC found were serotyped as O26:H11 (patient) and O119:H7 (control). Antimicrobial susceptibility tests revealed that 43.6%, 29.5% and 2.6% of the DEC isolates were resistant to ampicillin, cotrimoxazole and gentamicin, respectively. Our data indicate that EAEC remains prevalent among children living in Botucatu, and revealed atypical EPEC as emerging putative diarrheal agents in this geographical region.

Clinical and microbiologic evaluation of chronic leg ulcers: a cross-sectional study.

OBJECTIVE: This study aimed at evaluating the flora and bacterial load of chronic leg ulcers (CLUs) according to the clinical judgment of colonization or infection. DESIGN: This was an analytical and cross-sectional study. SETTING: This study was conducted in an outpatient wound care unit in the Dermatology Department of the Botucatu School of Medicine-UNESP, Brazil. PARTICIPANTS: The participants were patients with CLUs who did not use systemic antibiotics. METHODS: The ulcers were clinically divided into 3 groups: ulcers with good granulation tissue (GGT), critical colonization (CC), and infection. Secretion was collected from a 1-cm area using a swab and seeded by the semiquantitative method. OUTCOME MEASURES: The main outcome measures were genus and species of the bacteria found in the cultures and result of the semiquantitative culture correlating with the clinical diagnosis of GGT, CC, and infection. MAIN RESULTS: Seventy-seven ulcers were evaluated: 27 with GGT, 29 with CC, and 21 with infection. Gram-negative bacteria were most often found in all groups (81%): Pseudomonas aeruginosa, in granulation and colonized ulcers, and Proteus mirabilis, in infected ulcers. Ulcers from the infected group showed higher bacterial load. CONCLUSIONS: The flora of CLUs was predominantly constituted by gram-negative bacteria, and P aeruginosa was the most prevalent. The bacterial load of infected ulcers was higher as compared with the others, although some ulcers with GGT also presented a high load. The interpretation of microbiologic tests based on the swab techniques and even on semiquantitative analysis requires close clinical correlation.

Peritoneal dialysis-related peritonitis due to Staphylococcus aureus: a single-center experience over 15 years.

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 ß-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, ß-hemolysin production is predictive of non-resolution of infection, suggesting a pathogenic role of this factor in PD-related S. aureus peritonitis.

Oxacillin resistance and antimicrobial susceptibility profile of Staphylococcus saprophyticus and other staphylococci isolated from patients with urinary tract infection.

BACKGROUND: Staphylococcus saprophyticus is the second most frequent community-acquired causative agent of urinary tract infection (UTI). The objective of this study was to evaluate the susceptibility profile and resistance detection in Staphylococcus species. isolated from patients with UTI. MATERIALS AND METHODS: The isolates were investigated using the disk diffusion method, Vitek I system, E-test®, and detection of the mecA gene. RESULTS: Most isolates (76.2%) were resistant to oxacillin by the disk diffusion method, followed by those resistant to penicillin (72.2%). The oxacillin disk diffusion method, E-test, and Vitek I method showed higher sensitivity (94.4%) and lower specificity (28.9, 26.5, and 24.0%, respectively) than the cefoxitin disk diffusion test (sensitivity: 83.5%, specificity: 85.5%) for the detection of oxacillin resistance. CONCLUSIONS: The large number of oxacillin-resistant isolates indicates that the breakpoint value recommended by the Clinical and Laboratory Standards Institute may overestimate oxacillin resistance in S. saprophyticus. Thus, changes in these guidelines are necessary for the correct detection of this resistance.