Sumiko Sinto

An outbreak of multiresistant Acinetobacter baumanii in a university hospital in São Paulo, Brazil.

A case-control (46 cases, 23 controls) study was done to determine risk factors for an outbreak of a multiresistant Acinetobacter baumanii (only susceptible to colistin) in a university hospital. The use of antecedent antibacterials and intubation were independent risk factors. No common source was found. With control measures, the outbreak resolved gradually.

Nosocomial infections caused by multiresistant Pseudomonas aeruginosa.

A case-control study was done to evaluate factors associated with nosocomial infections by multiresistant Pseudomonas aeruginosa (MRPA). Results showed that MRPA was associated with the use of immunosuppressive and antimicrobial drugs. Five typing methods indicated that the MRPA infections were due to multiple strains rather than a single strain.

Involvement of pmrAB and phoPQ in polymyxin B adaptation and inducible resistance in non-cystic fibrosis clinical isolates of Pseudomonas aeruginosa.

ABSTRACT During investigation of susceptibility testing methods for polymyxins, 24 multidrug-resistant clinical isolates of Pseudomonas aeruginosa were observed to have a distinct, reproducible phenotype in which skipped wells were observed during broth microdilution testing for polymyxin B. Possible mechanisms underlying this phenotype were investigated. The effects of various concentrations of polymyxin B on growth, the expression of resistance genes, and outer-membrane permeability were observed. Real-time PCR was performed to compare the expression, in response to selected concentrations of polymyxin B, of genes related to the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems in polymyxin B-susceptible isolate PAO1, polymyxin B-resistant isolate 9BR, and two isolates (19BR and 213BR) exhibiting the skipped-well phenotype. 19BR and 213BR appeared to have similar basal levels of expression compared to that of PAO1 for phoQ, arnB, and PA4773 (from the pmrAB operon), and in contrast, 9BR had 52- and 280-fold higher expression of arnB and PA4773, respectively. The expression of arnB and PA4773 increased in response to polymyxin B in a concentration-dependent manner for 9BR but not for 19BR and 213BR. For these isolates, expression was significantly increased for arnB and PA4773, as well as phoQ, only upon exposure to 2 μg/ml polymyxin B but not at a lower concentration of 0.125 μg/ml. The sequencing of the pmrAB and phoPQ operons for all three isolates revealed a number of unique mutations compared to that for PAO1. 1-N-phenylnaphthylamine (NPN) was used to study the effect of preincubation with polymyxin B on the self-promoted uptake of polymyxin B across the outer membrane. The preincubation of cells with 2 μg/ml polymyxin B affected baseline membrane permeability in 19BR and 213BR and also resulted in a reduced rate of NPN uptake in these isolates and in PAO1 but not in 9BR. The results presented here suggest that the skipped-well isolates have the ability to adapt to specific concentrations of polymyxin B, inducing known polymyxin B resistance genes involved in generating alterations in the outer membrane.

Detection of mycobacteria in the bloodstream of patients with acquired immunodeficiency syndrome in a university hospital in Brazil.

This study was done to determine the occurrence of mycobacteria in the bloodstreams of patients with fever and advanced AIDS in a Brazilian hospital. We also verified the capability of an automated method for recovering these bacteria. During a period of 19 months, 254 patients with AIDS were evaluated. Blood cultures were generally submitted in pairs and drawn separately. Blood cultures were processed by the BACTEC 460TB System (Becton Dickinson Microbiology Systems, Sparks, MD), using the Bactec 13A media (Becton Dickinson Microbiology Systems, Sparks, MD). Of the 530 vials submitted, 77 (14.5%) from 41 (16%) patients were positive. Mycobacterium avium complex was recovered from 45 (58.4%) of the 77 positive vials, corresponding to 22 (53.6%) patients with positive blood cultures. The average time to detect Mycobacterium avium complex was 15 days. Mycobacterium tuberculosis was recovered from 26 (33.8%) of the 77 positive vials, corresponding to 15 (36.6%) patients with positive blood cultures, with an average detection time of 24 days. Other species of mycobacteria were recovered from 6 (7.8%) of the 77 vials, corresponding to 4 (9.8%) patients. M.avium complex was fairly prevalent (8.7%) in severely ill patients with AIDS in our hospital. M. tuberculosis was also an important (6.0%) agent of systemic bacterial infections in these patients. The rapid diagnosis of mycobacteremia was possible with the implementation of this automated technology.

Proteases (caseinase and elastase), hemolysins, adhesion and susceptibility to antimicrobials of Stenotrophomonas maltophilia isolates obtained from clinical specimens

Forty-six S. maltophilia isolates obtained from hospital clinical specimens were studied for protease (caseinase and elastase) production, hemolytic activity, adhesion to HEp-2 cells, plastic and glass. Susceptibility to antimicrobial agents was also evaluated. The majority of isolates were obtained from respiratory tract secretions of patients using medical devices. All the isolates grown overnight were able to hydrolyze casein at 30oC and 37oC. After 72h, all the isolates hydrolyzed elastase at 30oC and 40 isolates (87%) at 37oC. Most of the isolates presented hemolytic activity after 96h of incubation at both temperatures. Rabbit blood showed the hightest hemolytic activity, after 96h 61% and 98% of tested isolates presented b-hemolysis at 30oC and 37oC, respectively. All isolates were susceptible to trimethoprim-sulfametoxazole and were resistant to most b-lactams tested. By the dilution method, S. maltophilia showed a high susceptibility to ticarcillin-clavulanate and a lower susceptibility to ciprofloxacin than the agar diffusion. The isolates showed adhesion to HEp-2 cells, plastic and glass. The proteolytic activities and adhesion to inanimate surfaces detected in S. maltophilia can be related to the pathogenesis of this bacterium and/or medical device colonization which favors the development of nosocomial infections.

Pseudomonas aeruginosa clonal dissemination in Brazilian intensive care units

OBJECTIVE Investigate clonal dissemination of nosocomial multidrug-resistant Pseudomonas aeruginosa isolates within and between Brazilian intensive care units, which participated in the MYSTIC Program Brazil 2002. METHODS Thirty-six P. aeruginosa isolates resistant to meropenem or imipenem plus at least two of the following drugs: ciprofloxacin, cefepime, ceftazidime or piperacillin/tazobactam were isolated during 2002 at 4 centres in São Paulo and 1 centre in Brasília. Chromosomal restriction fragments obtained with SpeI were separated by pulsed-field gel electrophoresis (PFGE). Electrophoretic patterns were analyzed with GelCompar II v. 2.5. RESULTS Five major clones were identified (A, B, C, D, G). Clone A was constituted by 8 isolates with indistinguishable PFGE pattern present in 2 centres. Clone B was constituted by 4 indistinguishable isolates predominant in centre 6. Clone C had 3 indistinguishable isolates, with closely related clones (C1-3). Also, Clone D had 3 indistinguishable isolates, with closely related (D1) and possibly related (D2/D3) clones. Clones C and D were present in centre 1. Clone G was constituted by 2 indistinguishable isolates and was present in centre 7. Finally, 8 isolates were unique. Isolates from Centre 4 were unique. CONCLUSIONS Clonal dissemination was detected within (clones A, B, C, D, and G) and between centres (clone A). These findings are important when analyzing surveillance data, since susceptibility rates may be significantly affected by the dissemination of a resistant clone.

Atividade antimicrobiana in vitro de quinupristina/dalfopristina para cocos gram-positivos isolados de cinco centros brasileiros: resultado do estudo de vigilância L-SMART

A progressive increase of resistance among Gram-positive cocci (GPC) towards some antimicrobial agents has been observed for the past few years. This rise of resistance, most often seen in Staphylococcus spp., and Enterococcus spp., has mainly been noticed in hospital environments. Due to these recent patterns of resistance, newly developed antimicrobial agents need to be evaluated for the treatment of infections caused by these multi-resistant microorganisms. Quinupristin/dalfopristin (Q/D), also known as Synercid®, is an antimicrobial agent of intravenous administration, composed of two semi-synthetic derivatives of pristinomycin belonging to the group of streptogramins. The combination of streptogramins B and A at 3:7 ratio has an antimicrobial activity against gram-positive cocci. This combination has potent activity against gram-positive cocci such as Staphylococcus spp., Streptococcus spp. including Streptococcus pneumoniae, and Enterococcus faecium. However, strains of E. faecalis are usually resistant to this compound. The aim of this study was to evaluate the in vitro activity of Q/D and other eight antibiotics against 631 strains of GPC isolated from five Brazilian centers. Additionally, 20 vancomycin-resistant strains of E. faecium provided by a reference center from the United States were also included in this study. Minimal Inhibitory Concentrations (MICs) were determined by E-test methodology (AB Biodisk, Solna, Sweden), using standardized and controlled procedures. The evaluated strains were as follows: Staphylococcus aureus (267), coagulase negative Staphylococcus (131), Streptococcus pneumoniae (130), b-hemolytic Streptococcus (28), Enterococcus faecalis (44), and E. faecium (51). Quinuprintin/dalfopristin presented an excellent activity against Staphylococcus spp., regardless if these were susceptible or not to oxacillin. Against S. pneumoniae, Q/D also presented excellent activitiy regardless of their susceptibility to penicillin. Among vancomycin susceptible E. faecium studied, the MIC90 was 3mg/ml where 45% of the strains were susceptible, and 55% revealed intermediate resistance to quinupristin/dalfopristin. Overall, Q/D showed good activity against Staphylococcus spp., Streptococcus spp. including S. pneumoniae, and Enterococcus faecium representing a new option for the treatment of infections caused by multi-resistant gram-positive cocci, as well as an alternative for the use of glycopeptides.

Evaluation of the use and re-use of cotton fabrics as medical and hospital wraps

The objective of this study was to verify the efficacy of cotton fabric, made of serge bonding 2 x 1, as microbial barrier, when new and after multiple laundering and steam sterilization procedures. The power of the microbial barrier was correlated with physical characteristics of the fabric, using standard test methods for evaluation of weight, traction, stretching tearing resistance and microbiological characteristics. The microbiological results evidenced that the microbial barrier was effective when the wrapping material was new or went through a maximum of new 65 reprocessing procedures. As for the alterations in the physical characteristics of the reprocessed material, the decrease in weight seemed to be the event responsible for microbial barrier breaking. The timing of detected alterations in bursting, traction and stretching in the wrap and the reprocessed fabrics did not coincide with the moment of bacterial barrier breaking. The present investigation corroborates that the use double cotton fabric, for wrapping medical and hospital items for steam sterilization, is safe. Re-use number must be controlled, not exceeding 65 times.

In vitro activity of fluoroquinolones (gatifloxacin, levofloxacin and trovafloxacin) and seven other antibiotics against Streptococcus pneumoniae

In recent years, the level of resistance of S. pneumoniae to beta-lactam and/or macrolides has increased around the world including some countries in South America. Because of this resistance, it is necessary to test the therapeutic alternatives for treating this pathogen, including the newer quinolones. This study was carried out in order to compare the in vitro activity of fluoroquinolones gatifloxacin, levofloxacin and trovafloxacin, to penicillin G, amoxicillin, amoxicillin-clavulanate, cufuroxime sodium, ceftriaxone, azithromycin and clarithromycin, against 300 strains of S. pneumoniae. Of the 300 samples tested, 18.6% were not susceptible to penicillin (56 strains) and 7% (21 strains) were resistant to the second generation cephalosporin. Among the macrolides, resistance ranged from 6.7% for clarithromycin to 29.6% for azithromycin. Susceptibility to the newer quinolones was 100% including the 56 strains not susceptible to penicillin. Among the 10 antibiotics evaluated, the fluoroquinolones gatifloxacin, levofloxacin, and trovafloxacin displayed high levels of in vitro activity against S. pneumoniae.

Epidemiologic investigation of an outbreak of coagulase-negative Staphylococcus primary bacteremia in a newborn intensive care unit

Infections due to coagulase-negative Staphylococcus (CNS) are an ever-increasing nosocomial problem, particularly in the pediatric population. The authors describe a cluster of three primary bloodstream infections due to CNS in a newborn intensive care unit that occurred between November 23 and December 2, 1992. Two children died as a direct consequence of the bacteremia; at autopsy, one had a large bacteria-containing thrombus extending from the insertion site of a central catheter to the superior vena cava. The children were placed in isolation, and the nursing and medical staff were given topical nasal mupirocin. Plasmid analysis performed later disclosed three different blood isolates that also were different from any of the staffs nasal isolates. The authors concluded that molecular methods such as plasmid analysis are important tools in identifying true outbreaks and can prevent needless interventions, such as those during this cluster.