Ana Catarina de Souza Lopes

First report of KPC-producing Pseudomonas aeruginosa in Brazil.

The first report of KPC-producing Pseudomonas aeruginosa isolates was from Colombia (KPC-2) in 2007, followed by reports from Puerto Rico (KPC), Trinidad and Tobago (KPC-2), the United States (KPC-2), and China (KPC-2) ([1][1], [6][2][–][3][10][4]). This report describes the first detection of KPC

Phenotypic and molecular characterization of antimicrobial resistance and virulence factors in Pseudomonas aeruginosa clinical isolates from Recife, State of Pernambuco, Brazil

INTRODUCTION The emergence of carbapenem resistance mechanisms in Pseudomonas aeruginosa has been outstanding due to the wide spectrum of antimicrobial degradation of these bacteria, reducing of therapeutic options. METHODS Sixty-one clinical strains of P. aeruginosa isolated from five public hospitals in Recife, Pernambuco, Brazil, were examined between 2006 and 2010, aiming of evaluating the profiles of virulence, resistance to antimicrobials, presence of metallo-β-lactamase (MBL) genes, and clonal relationship among isolates. RESULTS A high percentage of virulence factors (34.4% mucoid colonies; 70.5% pyocyanin; 93.4% gelatinase positives; and 72.1% hemolysin positive) and a high percentage of antimicrobial resistance rates (4.9% pan-resistant and 54.1% multi-drug resistant isolates) were observed. Among the 29 isolates resistant to imipenem and/or ceftazidime, 44.8% (13/29) were MBL producers by phenotypic evaluation, and of these, 46.2% (6/13) were positive for the blaSPM-1 gene. The blaIMP and blaVIM genes were not detected. The molecular typing revealed 21 molecular profiles of which seven were detected in distinct hospitals and periods. Among the six positive blaSPM-1 isolates, three presented the same clonal profile and were from the same hospital, whereas the other three presented different clonal profiles. CONCLUSIONS These results revealed that P. aeruginosa is able to accumulate different resistance and virulence factors, making the treatment of infections difficult. The identification of blaSPM-1 genes and the dissemination of clones in different hospitals, indicate the need for stricter application of infection control measures in hospitals in Recife, Brazil, aiming at reducing costs and damages caused by P. aeruginosa infections.

Prevalence of the bla (SHV) gene in Klebsiella pneumoniae isolates obtained from hospital and community infections and from the microbiota of healthy individuals in Recife, Brazil.

The aim of this study was to determine the prevalence of the blaSHV gene in Klebsiella pneumoniae isolates from hospital and community infections and from the normal microbiota of healthy individuals in Recife, PE, Brazil. Fifty-two K. pneumoniae isolates were analyzed regarding the presence of the blaSHV gene, using PCR, and eight isolates were analyzed by DNA sequencing. This gene was detected in 16 isolates from hospital infections, four from community infections, and nine from the normal microbiota. This was the first study to find the blaSHV gene in K. pneumoniae isolates from the normal microbiota. Through DNA sequencing of eight K. pneumoniae isolates from hospital and community infections, with a resistance phenotype indicative of extended-spectrum β-lactamase production, a new SHV variant named SHV-122 was found. We also detected the presence of blaSHV-1, blaSHV-11, blaSHV-28, and blaSHV-108. The results show that in Recife, Brazil, K. pneumoniae isolates that presented resistance to oxyimino-β-lactams had high prevalence and diversity of the blaSHV gene. We also conclude that there was a high presence of the blaSHV gene among isolates from the normal microbiota of healthy individuals.

Phylogenetic Groups Among Klebsiella pneumoniae Isolates from Brazil: Relationship with Antimicrobial Resistance and Origin

The objectives of this study were to determine the distribution of phylogenetic groups among Klebsiella pneumoniae isolates from Recife, Brazil and to assess the relationship between the groups and the isolation sites and resistance profile. Ninety four isolates of K. pneumoniae from hospital or community infections and from normal microbiota were analyzed by gyrA PCR–RFLP, antibiotic susceptibility, and adonitol fermentation. The results revealed the distinction of three phylogenetic groups, as it has also been reported in Europe, showing that these clusters are highly conserved within K. pneumoniae. Group KpI was dominantly represented by hospital and community isolates while groups KpII and KpIII displayed mainly normal microbiota isolates. The resistance to third generation cephalosporins, aztreonam, imipenem, amoxicillin/clavulanic acid, and streptomycin was only observed in KpI. The percentage of resistance was higher in KpI, followed by KpII and KpIII. The differences in the distribution of K. pneumoniae phylogenetic groups observed in this study suggest distinctive clinical and epidemiological characteristics among the three groups, which is important to understand the epidemiology of infections caused by this organism. This is the first study in Brazil on K. pneumoniae isolates from normal microbiota and community infections regarding the distribution of phylogenetic groups based on the gyrA gene.

Colonização pelo Staphylococcus aureus em profissionais de enfermagem de um hospital escola de Pernambuco

Estudio realizado para identificar prevalencia de colonizacion por Staphylococcus aureus en profesionales de enfermeria de hospital universitario de Pernambuco, asi como evaluar el perfil de resistencia de la bacteria aislada. Se realizo un estudio transversal en el que se recolectaron muestras biologicas de manos y cavidad nasal. La identificacion del S. aureus se realizo mediante cultivo en agar-sangre, agar-manitol salado y mediante pruebas de catalasa y coagulasa. El perfil de sensibilidad se determino por tecnica de Kirby Bauer y para la determinacion de resistencia a meticilina se realizo screening en placa con oxalacina, con adicion de 4% de NaCl. De 150 profesionales evaluados, 39 estaban colonizados, lo que demostro prevalencia de 25,8%. Entre las variables estudiadas, faja etaria y cantidad de EPI se presentaron asociadas con la colonizacion por la bacteria. De todas las cepas aisladas, apenas cinco presentaron resistencia a meticilina.This study was performed with the objective to identify the prevalence of colonization by Staphylococcus aureus in nursing professionals from a teaching hospital in Pernambuco, and evaluate the resistance profile of these isolates. To do this, we performed a cross-sectional study where biological samples were collected from the hands and nasal cavities of the subjects. S. aureus was identified using agar (blood agar and mannitol salt) via catalase and coagulase tests. The sensitivity profile was determined by Kirby Bauer technique and determination of methicillin resistance was performed with oxacillin screening with sodium chloride (NaCl) addition. Of the 151 professionals evaluated, 39 were colonized which showed a prevalence of 25.8%. Among the variables studied, age and use of PPE were associated with colonization by the organism. Of all the isolates, only five were resistant to methicillin.

Presence of fimH, mrkD, and irp2 Virulence Genes in KPC-2-Producing Klebsiella pneumoniae Isolates in Recife-PE, Brazil

Klebsiella pneumoniae strains can produce different virulence factors, such as fimbrial adhesins and siderophores, which are important in the colonization and development of the infection. The aims of this study were to determine the occurrence of fimH, mrkD, and irp2 virulence genes in 22 KPC-2-producing K. pneumoniae isolates as well as 22 not producing-KPC isolates, from patients from different hospitals in Recife-PE, Brazil, and also to analyze the clonal relationship of the isolates by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The genes were detected by PCR and DNA sequencing. The blaKPC-2 gene was identified in 22 KPC-positive isolates. On analyzing the antimicrobial susceptibility profile of the isolates, it was detected that polymyxin and amikacin were the antimicrobials of best activity against K. pneumoniae. On the other hand, five isolates exhibited resistance to polymyxin. In the KPC-positive group, was observed a high rate of resistance to cephalosporins, followed by carbapenems. Molecular typing by ERIC-PCR detected 38 genetic profiles, demonstrating a multiclonal spread of the isolates analyzed. It was observed that the virulence genes irp2, mrkD, and fimH were seen to have together a higher frequency in the KPC-positive group. The accumulation of virulence genes of KPC-positive K. pneumoniae isolates, observed in this study, along with the multi-resistance impose significant therapeutic limitations on the treatment of infections caused by K. pneumoniae.

Colonization by Staphylococcus aureus among the nursing staff of a teaching hospital in Pernambuco

Estudio realizado para identificar prevalencia de colonizacion por Staphylococcus aureus en profesionales de enfermeria de hospital universitario de Pernambuco, asi como evaluar el perfil de resistencia de la bacteria aislada. Se realizo un estudio transversal en el que se recolectaron muestras biologicas de manos y cavidad nasal. La identificacion del S. aureus se realizo mediante cultivo en agar-sangre, agar-manitol salado y mediante pruebas de catalasa y coagulasa. El perfil de sensibilidad se determino por tecnica de Kirby Bauer y para la determinacion de resistencia a meticilina se realizo screening en placa con oxalacina, con adicion de 4% de NaCl. De 150 profesionales evaluados, 39 estaban colonizados, lo que demostro prevalencia de 25,8%. Entre las variables estudiadas, faja etaria y cantidad de EPI se presentaron asociadas con la colonizacion por la bacteria. De todas las cepas aisladas, apenas cinco presentaron resistencia a meticilina.This study was performed with the objective to identify the prevalence of colonization by Staphylococcus aureus in nursing professionals from a teaching hospital in Pernambuco, and evaluate the resistance profile of these isolates. To do this, we performed a cross-sectional study where biological samples were collected from the hands and nasal cavities of the subjects. S. aureus was identified using agar (blood agar and mannitol salt) via catalase and coagulase tests. The sensitivity profile was determined by Kirby Bauer technique and determination of methicillin resistance was performed with oxacillin screening with sodium chloride (NaCl) addition. Of the 151 professionals evaluated, 39 were colonized which showed a prevalence of 25.8%. Among the variables studied, age and use of PPE were associated with colonization by the organism. Of all the isolates, only five were resistant to methicillin.

Phenotypic and molecular characterization of resistance to macrolides, lincosamides and type B streptogramin of clinical isolates of Staphylococcus spp. of a university hospital in Recife, Pernambuco, Brazil

INTRODUCTION There is a mechanism of macrolide resistance in Staphylococcus spp. which also affects the lincosamides and type B streptogramins characterizing the so-called MLSB resistance, whose expression can be constitutive (cMLSB) or inducible (iMLSB) and is encoded mainly by ermA and ermC genes. The cMLSB resistance is easily detected by susceptibility testing used in the laboratory routine, but iMLSB resistance is not. Therapy with clindamycin in cases of infection with isolated iMLSB resistance may fail. OBJECTIVE To characterize the phenotypic (occurrence of cMLSB and iMLSB phenotypes) and molecular (occurrence of ermA and ermC genes) profiles of MLSB resistance of clinical isolates of susceptible and methicillin-resistant Staphylococcus aureus and CNS (coagulase-negative Staphylococcus) from patients of a university hospital, in Pernambuco. METHODS The antimicrobial susceptibility of 103 isolates was determined by the disk diffusion technique in Mueller-Hinton agar followed by oxacillin screening. The iMLSB phenotype was detected by D test. Isolates with cMLSB and iMLSB phenotypes were subjected to polymerase chain reaction (PCR) for the detection of ermA and ermC genes. RESULTS The cMLSB and iMLSB phenotypes were respectively identified in 39 (37.9%) and five (4.9%) isolates. The iMLSB phenotype was found only in four (10.8%) methicillin-susceptible S. aureus and one (4.5%) methicillin-resistant S. aureus. In the 44 isolates subjected to PCR, four (9.1%) only ermA gene was detected, a lower frequency when compared to only ermC 17 (38.6%) gene and to one (2.3%) isolate presenting both genes. CONCLUSION In the Staphylococcus spp. analyzed, the ermC gene was found more often than the ermA, although the iMLSB phenotype had been less frequent than the cMLSB. It was important to perform the D test for its detection to guide therapeutic approaches.

Characterization of the bacterial microbiota of Biomphalaria glabrata (Say, 1818) (Mollusca: Gastropoda) from Brazil.

Roughly 200 000 000 people in 74 countries infected with schistosomes all share the fact that they came in contact freshwater harbouring infected snails. The aim of the study is to characterize the microbiota of wild and laboratory‐reared snails of Biomphalaria glabrata from Pernambuco, Brazil. The microbiota of these molluscs was identified biochemically by the VITEK 2 automated microbiological system. Antimicrobial susceptibility testing was carried out by the disc diffusion method with ß‐lactam antibiotics, aminoglycosides, quinolones, folate pathway inhibitors, fenicols and tetracyclines. The results showed that all bacteria identified were gram‐negative, including 11 bacterial genera: Aeromonas, Citrobacter, Enterobacter, Cupriavidus, Rhizobium, Stenotrophomonas, Pseudomonas, Klebsiella, Acinetobacter, Vibrio and Sphingomonas. Regarding the antimicrobial susceptibility, all the isolates exhibited resistance to amoxicillin and sensitivity to meropenem (beta‐lactam antimicrobials). The microbiota of the wild snails consisted predominantly of Enterobacter cloacae, while the laboratory‐reared snails predominantly showed Citrobacter freundii and Aeromonas sobria.

Análise epidemiológica de isolados clínicos de Pseudomonas aeruginosa provenientes de hospital universitário

OBJECTIVES Pseudomonas aeruginosa is an increasingly prevalent opportunistic pathogen in hospital infection cases. Its high resistance rates to many antimicrobials has given this microorganism a relevant role among other highly prevalent bacteria involved in nosocomial infections. This study aimed to analyze epidemiologic characteristics of P. aeruginosa and to evaluate its susceptibility to antimicrobial agents at Hospital das Clínicas of the Universidade Federal de Pernambuco METHODS A retrospective study was performed based on the registry book of miscellaneous secretions from the bacteriology laboratory of the Hospital das Clínicas involving the period between January and June 2008. Among the secretions registered, were identified the positives samples for P. aeruginosa, whose origin was analyzed, as well as its susceptibility profile to routinely used in our laboratory antimicrobials. RESULTS The bacteria most frequently isolated from miscellaneous secretions bacteria were P. aeruginosa (26%) and S. aureus (25%). P. aeruginosa was mainly isolated from respiratory infections, with 33% of positive samples for this organism from tracheal secretions and 21% from nasal. The most effective antimicrobials against P. aeruginosa were: amikacin, imipenem, meropenem and aztreonam. CONCLUSIONS These results show a high prevalence of P. aeruginosa in the Hospital das Clínicas of the Universidade Federal de Pernambuco. Despite featuring high resistance rates to older antimicrobials, as cephalosporins first and second generations and chloramphenicol, this pathogen showed good susceptibility to agents routinely used in this hospital.OBJETIVOS: A Pseudomonas aeruginosa e um patogeno oportunista que tem se destacado quanto a prevalencia em casos de infeccoes hospitalares. Sua ampla resistencia aos diversos grupos de antimicrobianos garante a este microrganismo um papel de destaque entre as bacterias mais prevalentes associadas a infeccao nosocomial. O objetivo deste estudo foi realizar um levantamento epidemiologico da P. aeruginosa, bem como do seu perfil de susceptibilidade aos antimicrobianos no Hospital das Clinicas da Universidade Federal de Pernambuco. METODOS: Foi realizado um estudo retrospectivo baseado no livro de registro de secrecoes diversas do laboratorio de bacteriologia do Hospital das Clinicas no periodo compreendido entre janeiro a junho de 2008. Entre os registros, identificamos aqueles que foram positivos para a P. aeruginosa, analisando sua origem e perfil de susceptibilidade aos antimicrobianos utilizados na rotina daquele laboratorio. RESULTADOS: As bacterias mais frequentes, isoladas das secrecoes diversas, foram P. aeruginosa (26%) e S. aureus (25%). Quanto a origem, a P. aeruginosa foi isolada principalmente de infeccoes respiratorias, pois 33% das amostras positivas para esta bacteria foram provinientes de secrecoes traqueais e 21% nasais. Os antimicrobianos mais eficazes contra a P. aeruginosa foram: amicacina, imipenem, meropenem e aztreonam. CONCLUSOES: Estes resultados mostram uma alta prevalencia de P. aeruginosa, no Hospital das Clinicas da Universidade Federal de Pernambuco. Apesar de apresentar grande resistencia a antimicrobianos mais antigos como as cefalosporinas de primeira e segunda geracao, assim como cloranfenicol, em geral, este patogeno demonstrou boa sensibilidade as drogas utilizadas na rotina deste hospital.