Risk factors for recovery of Imipenem-or ceftazidime-resistant Pseudomonas aeruginosa among patients admitted to a teaching hospital in Brazil

background. The prevalence of resistance to imipenem and ceftazidime among Pseudomonas aeruginosa isolates is increasing worldwide.objective. Risk factors for nosocomial recovery ( defined as the finding of culture- positive isolates after hospital admission) of imipenemresistant P. aeruginosa ( IRPA) and ceftazidime- resistant P. aeruginosa ( CRPA) were determined.design. Two separate case- control studies were conducted. Control subjects were matched to case patients ( ratio, 2: 1) on the basis of admission to the same ward at the same time as the case patient. Variables investigated included demographic characteristics, comorbid conditions, and the classes of antimicrobials used.setting. The study was conducted in a 400- bed general teaching hospital in Campinas, Brazil that has 14,500 admissions per year. Case patients and control subjects were selected from persons who were admitted to the hospital during 1992 - 2002.results. IRPA and CRPA isolates were obtained from 108 and 55 patients, respectively. Statistically significant risk factors for acquisition of IRPA were previous admission to another hospital ( odds ratio [ OR], 4.21 [ 95% confidence interval {CI}, 1.40- 12.66];), hemodialysis Pp. 01 ( OR, 7.79 [ 95% CI, 1.59- 38.16];), and therapy with imipenem ( OR, 18.51 [ 95% CI, 6.30- 54.43];), amikacin ( OR, 3.22 Pp. 01 P !.001 [ 95% CI, 1.40- 7.41];), and/ or vancomycin ( OR, 2.48 [ 95% CI, 1.08- 5.64];). Risk factors for recovery of CRPA were Pp. 005 Pp. 03 previous admission to another hospital ( OR, 18.69 [ 95% CI, 2.00- 174.28];) and amikacin use ( OR, 3.69 [ 95% CI, 1.32- 10.35]; Pp. 01). Pp. 01conclusion. Our study suggests a definite role for several classes of antimicrobials as risk factors for recovery of IRPA but not for recovery of CRPA. Limiting the use of only imipenem and ceftazidime may not be a wise strategy to contain the spread of resistant P. aeruginosa strains.

Evaluation of the antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolated from human infections

Multiresistant Staphylococcus aureus constitutes an important public health problem, especially in view of its possible spread in nosocomial environments. In the present work, we analyzed the susceptibility profile of 80 S. aureus stains from human infections resistant to at least 10 drugs. For this study, the techniques used were the disk method and minimum inhibitory concentration (MIC) of the following drugs: cefuroxime, ciprofloxacin, clindamycin, erythromycin, gentamycin, imipenem, oxacillin, rifampicin, tetracycline and vancomycin, according the criteria of the National Committee for Clinical Laboratory Standards (NCCLS). Methicillin was included in the antibiogram as a marker, which is usually used in drugs selection for the treatment of staphylococcal infections. Results indicated that the most effective drug was vancomycin. For the other 10 drugs, the percentage of resistant strains ranged from 85% to 93.75%. In relation to the MICs, it was observed that vancomycin (MIC 90% = 0.615ug/ml) was the most effective drug; followed by rifampicin (MIC 90% = 2.6ug/ml) and ciprofloxacin (MIC 90% = 26.6ug/ml). The drugs that showed the least effective activity were cefuroxime, clindamycin, erythromycin, gentamycin, and oxacillin. On the other hand, observation of β-lactamase production revealed that most of the methicillin-resistant strains produced β-lactamase (83.7%), potentiating the risks of nosocomial infections. In general, vancomycin still continues to be one of the most effective drugs for staphylococcal infections therapy.

In vitro antimicrobial activity of acroseal, polifl and epiphany against Enterococcus faecalis

Using the agar diffusion method, this study evaluated the in vitro antimicrobial activity of the commercial endodontic sealers Acroseal and Epiphany, a castor-oil based experimental sealer, Polifl, and a primer agent (Epiphany self-etching primer), against Enterococcus faecalis. Zinc oxide and eugenol cement (ZOE) served as control. Five wells per dish were made at equidistant points and immediately flled with the test and control materials. After incubation of the dishes at 37°C for 24 h and 48 h, the diameter of the zones of microbial growth inhibition produced around the wells was measured (in mm) with a millimeter rule. After 48 h, the diameters of the zones of microbial growth inhibition were the same as those observed at 24 h, only the substances continued to diffuse. Epiphany and Polifl did not show antibacterial activity (no formation of zones of microbial growth inhibition). The primer produced the largest zones of inhibition (17.62 mm) followed by Acroseal (7.25 mm) and ZOE (7.12 mm). E. faecalis was resistant to Epiphany and Polifl, while the primer and Acroseal sealer were effective against this microorganism under the tested conditions.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a burn unit from Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) poses a threat for patients in burn units. Studies that mix epidemiological designs with molecular typing may contribute to the development of strategies for MRSA control. We conducted a study including: molecular characterization of Staphylococcal Chromosome Cassette mecA (SCCmec), strain typing with pulsed field gel electrophoresis (PFGE) and detection of virulence genes, altogether with a case-case-control study that assessed risk factors for MRSA and for methicillin-susceptible S. aureus (MSSA), using S. aureus negative patients as controls. Strains were collected from clinical and surveillance cultures from October 2006 through March 2009. MRSA was isolated from 96 patients. Most isolates (94.8%) harbored SCCmec type III. SCCmec type IV was identified in isolates from four patients. In only one case it could be epidemiologically characterized as community-associated. PFGE typing identified 36 coexisting MRSA clones. When compared to MSSA (38 isolates), MRSA isolates were more likely to harbor two virulence genes: tst and lukPV. Previous stay in other hospital and admission to Intensive Care Unit were independent risk factors for both MRSA and MSSA, while the number of burn wound excisions was significantly related with the former (OR = 6.80, 95%CI = 3.54-13.07). In conclusion, our study found polyclonal endemicity of MRSA in a burn unit, possibly related to importing of strains from other hospitals. Also, it pointed out to a role of surgical procedures in the dissemination of MRSA strains. © 2013 Elsevier Ltd and ISBI. All rights reserved.

Identificação de Enterococcus spp. e avaliação da ocorrência de genes de resistência á vancomicina em amostras de pacientes de hospitais de manaus, AM

Pós-graduação em Fisiopatologia em Clínica Médica - FMB

Prevalência, Virulência e sensibilidade às terapias antimicrobianas das cepas de enterecoccus faecalis e enterococcus faecium isoladas de infecções endodônticas

Pós-graduação em Biopatologia Bucal - ICT

Microbiological Profile Resistant to Different Intracanal Medications in Primary Endodontic Infections

Introduction: This clinical study aimed to determine the microbiological profile resistant to different intracanal medications in primary endodontic infections by using both microbiological culture and the checkerboard DNA-DNA hybridization technique. Methods: Twenty primarily infected root canals were selected and then instrumented before being randomly divided into 2 groups according to the intracanal medications: calcium hydroxide (Ca[OH](2)) or Ca(OH)(2) + chlorhexidine (CHX). Samples were collected before and after root canal procedures, which consisted in submitting them to microbiological culture and processing them for checkerboard DNA-DNA hybridization. Results: No differences were found between the Ca(OH)(2) (99.98%) and Ca(OH)(2) + CHX groups (99.76%) regarding the median percentage values for the reduction of cultivable bacteria. The most frequently detected species were Capnocytophaga ochracea (70%) and Fusobacterium nucleatum ssp. vincentii (70%) in the initial samples. After instrumentation, the most frequently detected species were E. faecium (60%). After root canal treatments using either Ca(OH)(2) or Ca(OH)(2) + CHX as intracanal medications, the most frequently detected species were E nucleatum ssp. vincentii (90%) and Enterococcus faecium (40%), respectively. Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, this reduction was significantly greater in the Ca(OH)(2) + CHX group (P < .05). This difference was also observed when evaluating the total bacterial load (P < .05). Conclusions: The use of Ca(OH)(2) associated with CHX as an intracanal medication showed better results by acting on gram-positive and gram-negative microorganisms although such an action to eradicate enterococci should also be sought.

Bacteriocinogenic and virulence potential of Enterococcus isolates obtained from raw milk and cheese

Aims To provide molecular and phenotypical characterization of Enterococcus isolates obtained from raw milk and cheese, regarding their bacteriocinogenic and virulence activity. Methods and Results Forty-three bacteriocinogenic enterococci isolates were identified by 16s rDNA, fingerprinted by RAPD-PCR analysis and tested by PCR for the presence of genes for lantibiotics (lanM, lanB and lanC) and enterocins (entA, entB, entP, entL50AB and entAS48) and by phenotypical methods for bacteriocin production and inhibitory spectrum. Also, the virulence of the isolates was evaluated by PCR for genes gelE, hyl, asa1, esp, cylA, efaA, ace, vanA, vanB, hdc1, hdc2, tdc and odc and by phenotypical tests for gelatinase, lipase, DNAse and a- and beta-haemolysis. Most isolates (93.0%) harboured at least one lantibiotic or enterocin gene and were positive for several tested virulence genes, mainly asa1 (100%), gelE (93.0%) and efaA (83.7%). 53.5% of the isolates presented beta-haemolysis. Conclusions Enterococcus spp. isolates presented an interesting potential application for food preservation because of bacteriocin production; however, virulence-related genes were identified in all RAPD profiles. Significance and Impact of the Study The study demonstrated the contradictory characteristics of the tested Enterococcus isolates: they presented a good potential for application in food biopreservation but contained several virulence factors.

Adverse events related to vancomycin use in a University Hospital in Brazil

Introduction and Objectives: Vancomycin is indicated to patients who have not responded to treatment with other antibiotics in serious infections caused by organisms susceptible to it and resistant to other antimicrobials. However, over the last five years, many adverse reactions have been reported with this medicine in the University Hospital of the University of Silo Paulo (HU/USP), such as nephrotoxicity and toxicity related to infusion. Some critical patients, for example surgical patients with sepsis and severe trauma are generally susceptible to renal failure due to the severity of the underlying disease. The-aim of this study is to quantify and delineate the epidemiological profile of confirmed adverse reactions caused by vancomycin. Material and Methods: We conducted a retrospective observational quantitative study of medical records of patients who had confirmed.adverse reactions occurred with vancomycin in the period from January 2007 to May 2012, at the HU/USP - Brazil. All notifications related to vancomycin were evaluated in the following items: age and sex of patients, type and ward where the adverse event occurred involving this drug. Results and Conclusions: During the analysed period, were confinued 37 adverse events with vancomycn. The adults represented 75,7% of the cases, and the children 24,3%. The present study shows that adult patients admitted to the medical clinic had greater susceptibility to adverse reactions to vancomycin and for pediatric patients its higher frequency was at ICU. Despite the adverse skin reactions performed with greater frequency, it is known that the most severe reactions were related to the kidney resulting in more complex clinical interventions.

Characterisation of probiotic strains for the control and prevention of enteropathogens in the food chain

60 strains (belonging to the genera Lactobacillus, Bifidobacterium, Leuconostoc and Enterococcus) were tested for their capacity to inhibit the growth of 3 strains of Campylobacter jejuni: Lactobacilli and bifidobacteria were left to grow in MRS or TPY broth at 37°C overnight in anaerobic conditions; Campylobacter jejuni was inoculated in blood agar plates at 37°C for 24-48 hours in microaerophilic conditions. The inhibition experiments were carried out in vitro using ”Spot agar test” and “Well diffusion assay” techniques testing both cellular activity and that of the surnatant. 11 strains proved to inhibit the growth of Campylobacter jejuni. These strains were subsequently analised analised in order to evaluate the resistance to particular situations of stress which are found in the gastrointestinal tract and during the industrial transformation processes (Starvation stress, osmotic stress, heat stress, resistance to pH and to bile salts). Resistance to starvation stress: all strains seemed to resist the stress (except one strain). Resistance to osmotic stress: all strains were relatively resistant to the concentrations of 6% w/v of NaCl (except one strain). Resistance to heat stress: only one strain showed little resistance to the 55°C temperature. Resistance to pH: In the presence of a low pH (2.5), many strains rapidly lost their viability after approximately 1 hour. Resistance to bile salts: Except for one strain, all strains seemed to be relatively resistant to the 2% w/v concentration of bile salts. Afterward, strains were identified by using phenotipic and molecular techniques. Phenotipic identification was carried out by using API 50 CHL (bioMérieux) and API 20 STREP identification system (bioMérieux); molecular identification with species-specific PCR: the molecular techniques confirmed the results by phenotipic identification. For testing the antibiotic resistance profile, bacterial strains were subcultured in MRS or TPY broth and incubated for 18 h at 37°C under anaerobic conditions. Antibiotics tested (Tetracycline, Trimethoprim, Cefuroxime, Kanamycin, Chloramphenicol, Vancomycin, Ampycillin, Sterptomycin, Erythromycin) were diluted to the final concentrations of: 2,4,8,16,32,64,128,256 mg/ml. Then, 20 μl fresh bacterial culture (final concentration in the plates approximately 106 cfu/ml) were added to 160 μl MRS or TPY broth and 20 μl antibiotic solution. As positive control the bacterial culture (20 ul) was added to broth (160 ul) and water (20 ul). Test was performed on plates P96, that after the inoculum were incubated for 24 h at 37oC, then the antibiotic resistance was determined by measuring the Optical Density (OD) at 620 nm with Multiscan EX. All strains showed a similar behaviour: resistance to all antibiotic tested. Further studies are needed.

Antimicrobial resistance of Escherichia coli and Enterococcus faecalis in housed laying-hen flocks in Europe

The aim of this study was to determine the potential association between housing type and multiple drug resistance (MDR) in Escherichia coli and Enterococcus faecalis isolates recovered from 283 laying-hen flocks. In each flock, a cloacal swab from four hens was collected and produced 1102 E. coli and 792 E. faecalis isolates. Broth microdilution was used to test susceptibility to antimicrobials. Country and housing type interacted differently with the MDR levels of both species. In the E. coli model, housing in a raised-floor system was associated with an increased risk of MDR compared to the conventional battery system [ odds ratio (OR) 2.12, 95% confidence interval (CI) 1.13-3.97)]. In the E. faecalis model the MDR levels were lower in free-range systems than in conventional battery cages (OR 0.51, 95% CI 0.27-0.94). In Belgium, ceftiofur-resistant E. coli isolates were more numerous than in the other countries.

Efficacy of telavancin against penicillin-resistant pneumococci and Staphylococcus aureus in a rabbit meningitis model and determination of kinetic parameters

The penetration of telavancin was 2% into inflamed meninges and ca. 1 per thousand into noninflamed meninges after two intravenous injections (30 mg/kg of body weight). In experimental meningitis, telavancin was significantly superior to vancomycin combined with ceftriaxone against a penicillin-resistant pneumococcal strain. Against a methicillin-sensitive staphylococcal strain, telavancin was slightly but not significantly superior to vancomycin.

Activities of ertapenem, a new long-acting carbapenem, against penicillin-sensitive or -resistant pneumococci in experimental meningitis

The penetration of ertapenem, a new carbapenem with a long half-life, reached 7.1 and 2.4% into inflamed and noninflamed meninges, respectively. Ertapenem had excellent antibacterial activity in the treatment of experimental meningitis due to penicillin-sensitive and -resistant pneumococci, leading to a decrease of 0.69 +/- 0.17 and 0.59 +/- 0.22 log(10) CFU/ml x h, respectively, in the viable cell counts in the cerebrospinal fluid. The efficacy of ertapenem was comparable to that of standard regimens (ceftriaxone monotherapy against the penicillin-sensitive strain and ceftriaxone combined with vancomycin against the penicillin-resistant strain). In vitro, ertapenem in concentrations above the MIC was highly bactericidal against both strains. Even against a penicillin- and quinolone-resistant mutant, ertapenem had similar bactericidal activity in vitro.

Gemifloxacin is efficacious against penicillin-resistant and quinolone-resistant pneumococci in experimental meningitis

In experimental rabbit meningitis, gemifloxacin penetrated inflamed meninges well (22 to 33%) and produced excellent bactericidal activity (change in log(10) [Deltalog(10)] CFU/ml/h, -0.68 +/- 0.30 [mean and standard deviation]), even superior to that of the standard regimen of ceftriaxone plus vancomycin (-0.49 +/- 0.09 deltalog(10) CFU/ml/h), in the treatment of meningitis due to a penicillin-resistant pneumococcal strain (MIC, 4 mg/liter). Even against a penicillin- and quinolone-resistant strain, gemifloxacin showed good bactericidal activity (-0.48 +/- 0.16 deltalog(10) CFU/ml/h). The excellent antibacterial activity of gemifloxacin was also confirmed by time-kill assays over 8 h in vitro.

Cefepime is efficacious against penicillin- and quinolone-resistant pneumococci in experimental meningitis

In experimental rabbit meningitis, cefepime given at a dose of 100 mg/kg was associated with concentrations in the cerebrospinal fluid of between 5.3 and 10 mg/L and a bactericidal activity of -0.61 +/- 0.24 Delta log(10) cfu/mL x h, similar to the standard regimen of ceftriaxone combined with vancomycin (-0.58 +/- 0.14 Delta log(10) cfu/mL x h) in the treatment of meningitis due to a penicillin- and quinolone-resistant pneumococcal mutant strain (MIC 4 mg/L). Compared with the penicillin-resistant parental strain, the penicillin- and quinolone-resistant mutant was killed more slowly by cefepime and ceftriaxone in time-killing assays in vitro over 8 h.