First report of bla GES-1 in Proteus mirabilis clinical isolates

Abstract Proteus mirabilis is one of the main pathogens causing urinary tract infections and sepsis. To our knowledge, this is the first report of a P. mirabilis hosting bla GES. The presence of these genes was determined using PCR and sequencing. We identified the presence of bla GES-1 in all three isolates. In addition, we identified the bla KPC-2 and bla NDM-1 genes in the two strains. These data emphasize the importance of monitoring and surveillance of all enterobacteria. The circulation of P. mirabilis strains carrying bla GES-1 constitutes a new scenario of resistance in this species and should be an epidemiological alert for global health.

Identification of plasmid IncQ1 and NTEKPC-IId harboring bla KPC-2 in isolates from Klebsiella pneumoniae infections in patients from Recife-PE, Brazil

Abstract INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.

Emergence of bla NDM-1 associated with the aac(6')-Ib-cr, acrB, cps, and mrkD genes in a clinical isolate of multi-drug resistant Klebsiella pneumoniae from Recife-PE, Brazil

Abstract INTRODUCTION: The emergence of New Delhi metallo-β-lactamase (NDM) is concernig because it reduces the antibiotic therapy options for bacterial infections. METHODS: Resistant and virulent genes from an isolate of Klebsiella pneumoniae derived from a patient with sepsis in a hospital in Recife-PE, Brazil, were investigated using PCR and DNA sequencing. RESULTS: bla NDM-1, aac(6')-Ib-cr and acrB resistance genes, and cps and mrkD virulence genes were detected. CONCLUSIONS To our knowledge, this is the first report on bla NDM-1 in Recife-PE. This detection alerts researchers to the need to control the spread of bla NDM-1 resistance gene by this bacterium in Brazil.

Relationships between phagocytosis, mucoid phenotype, and genetic characteristics of Klebsiella pneumoniae clinical isolates

Abstract INTRODUCTION The relationships between phagocytosis, and mucoid phenotype, plasmid profile and virulence, and resistance genetic characteristics of Klebsiella pneumoniae clinical isolates were evaluated. METHODS Thirty isolates were used to determine the mucoid aspect. Four were selected for analysis of phagocytosis by alveolar macrophages. RESULTS Thirty percent of the samples presented the mucoid phenotype. The phagocytosis rate ranged from 21.5% to 43.43%. Phagocytosis was not correlated with the plasmid profile, but was apparently correlated with mucoid phenotype and antibiotic susceptibility. CONCLUSIONS: Several virulence factors act in parallel in K. pneumoniae to impair host defense.

Detection of multidrug-resistant Pseudomonas aeruginosa harboring bla GES-1 and bla GES-11 in Recife, Brazil

Abstract INTRODUCTION: Pseudomonas aeruginosa, an important pathogen globally, presents several resistance mechanisms. This study aimed to investigate the presence of bla GES in clinical isolates of Pseudomonas aeruginosa obtained from various clinical specimens from patients admitted to three different hospitals in Recife, Brazil. The Guiana extended spectrum beta-lactamase (GES) enzymes are responsible for conferring broad spectrum resistance to beta-lactam drugs, including the carbapenems. METHODS: A total of 100 carbapenem-resistant P. aeruginosa isolates underwent polymerase chain reaction (PCR) testing to identify bla GES, bla KPC, bla SPM-1, bla IMP, and bla VIM. Additionally, PCR products positive for bla GES were sequenced. The clonal profiles of these same isolates were then determined by means of enterobacterial repetitive intergenic consensus (ERIC)-PCR analysis. RESULTS: PCR analysis revealed that four isolates harbored bla GES; DNA sequencing showed that two harbored bla GES-1 and two bla GES-11. Beta-lactamase genes bla SPM-1, bla IMP, bla VIM, and bla KPC were investigated; none of these genes was detected. Automated susceptibility testing methods (Vitek®2, bioMérieux) showed that the bla GES-1-positive isolates were only susceptible to polymyxin B. The patterns obtained with ERIC-PCR methods showed clonal relationship between the two isolates that harbored bla GES-11, whereas different clonal profiles were found in the isolates harboring bla GES-1. CONCLUSIONS: We detected the presence of bacterial isolates positive for two different variants of the enzyme GES in three different hospitals from Recife, Brazil. These enzymes have a great capacity for dissemination among Gram-negative bacteria and confer broad-spectrum resistance to beta-lactam antibiotics and to the carbapenems.

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

Abstract 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.

Colonização pelo Staphylococcus aureus em profissionais de enfermagem de um hospital escola de Pernambuco / Colonization by Staphylococcus aureus among the nursing staff of a teaching hospital in Pernambuco / Colonización por Staphylococcus aureus en profesionales de enfermería de un hospital escuela de Pernambuco

O presente estudo foi realizado com o objetivo de identificar a prevalência de colonização pelo Staphylococcus aureus em profissionais de enfermagem de um hospital universitário de Pernambuco, bem como avaliar o perfil de resistência deles isoladamente. Para isso, foi realizado um estudo transversal, no qual foram coletadas amostras biológicas das mãos e da cavidade nasal. A identificação do S. aureus foi realizada por meio do semeio em agar-sangue, agar manitol-salgado e através dos testes de catalase e coagulase. O perfil de sensibilidade foi determinado pela técnica de Kirby Bauer e para determinação da resistência à meticilina foi realizado o screening em placa com oxacilina com adição de 4% de NaCl. Dos 151 profissionais avaliados, 39 se encontravam colonizados, o que demonstrou uma prevalência de 25,8%. Dentre as variáveis estudadas, a faixa etária e a quantidade de EPI apresentaram-se associadas à colonização pelo microrganismo. De todas as linhagens isoladas, apenas cinco apresentaram resistência à 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.

Estudio realizado para identificar prevalencia de colonización por Staphylococcus aureus en profesionales de enfermería de hospital universitario de Pernambuco, así como evaluar el perfil de resistencia de la bacteria aislada. Se realizó un estudio transversal en el que se recolectaron muestras biológicas de manos y cavidad nasal. La identificación del S. aureus se realizó mediante cultivo en agar-sangre, agar-manitol salado y mediante pruebas de catalasa y coagulasa. El perfil de sensibilidad se determinó por técnica de Kirby Bauer y para la determinación de resistencia a meticilina se realizó screening en placa con oxalacina, con adición de 4% de NaCl. De 150 profesionales evaluados, 39 estaban colonizados, lo que demostró prevalencia de 25,8%. Entre las variables estudiadas, faja etaria y cantidad de EPI se presentaron asociadas con la colonización por la bacteria. De todas las cepas aisladas, apenas cinco presentaron resistencia a meticilina.

Epidemiological surveillance and susceptibility of Staphylococcus aureus among healthcare workers at a reference hospital: preliminary assessment / Vigilância epidemiológica e susceptibilidade de Staphylococcus aureus em profissionais de saúde de um hospital de referência: uma avaliação inicial

Healthcare professionals (HCPs) are liable to pathogenic microorganisms infection/colonization, which plays an important role as a potential source of transmission to patients, coworkers, relatives and communities.The present study evaluates the prevalence of Staphylococcus aureus colonization in HCPs who work at are ference hospital in Recife, PE, and the isolates profiles of susceptibility to antimicrobial drugs. A crosssectional study was undertaken, and HCPs from operating rooms, intensive care units (ICUs), hemodialysis and nephrology units of the Clinical Hospital of Pernambuco were evaluated. S. aureus isolates wereidentified by standard methods recommended by CLSI and the susceptibility to methicillin and vancomycin was determined by the minimum inhibitory concentration technique (E-test). The prevalence of S aureus observed among HCPs was 25.7%. Among S. aureus strains isolates, the highest percentage of antibioticresistance was observed in penicillin (91.4%), erythromycin (43.1%) and cefoxitin (17.2%). All of the strainswere sensitive to vancomycin. Three S. aureus methicillin-resistant (MRSA) strains were identified, whichwere isolated from the nursing aides staff. The prevalence of MRSA found in the present study was lowerthan those reported elsewhere. These findings suggest that a continuous assessment should be performedfor better understanding the dynamics of S. aureus colonization/infection in order to reduce the risks of infection by this microorganism.

Infecção/colonização por microrganismos patogênicos em profissionais de saúde (PS) representa uma potencial fonte de transmissão para os pacientes, colegas de trabalho, familiares e comunidade. No presente estudo foi avaliada a prevalência da colonização por Staphylococcus aureus em PS de um hospital de referência do Recife, no período de março e julho de 2007, e o perfil de susceptibilidade das cepas isoladas às drogas antimicrobianas. Foi realizado um estudo transversal, no qual os PS de salas cirúrgicas, unidades de terapiaintensiva (UTIs), hemodiálise e unidades de nefrologia foram avaliados. O isolamento e a identificação de S. aureus foram efetuados de acordo com as orientações do CLSI. O perfil de susceptibilidade da bactéria aos antimicrobianos meticilina e vancomicina foi determinado por meio de técnica de difusão em discoassociada à técnica de concentração inibitória mínima (E-test). A prevalência de colonização por S. aureus entre os PS foi de 25,7%. Entre as cepas isoladas de S. aureus, os maiores percentuais de resistência foram observados frente à penicilina (91,4%), eritromicina (43,1%) e cefoxitina (17,2%). Todos os isolados foram sensíveis à vancomicina. Três cepas foram identificadas como resistentes à meticilina, as quais foram isoladas de auxiliares de enfermagem. Sugere-se que avaliações contínuas sejam realizadas para melhor compreensão da dinâmica de colonização/infecção e redução dos riscos de infecção por esse microrganismo.

Análise epidemiológica de isolados clínicos de Pseudomonas aeruginosa provenientes de hospital universitário / Epidemiologic analysis of clinical isolates of Pseudomonas aeruginosa from an university hospital

OBJETIVOS: A Pseudomonas aeruginosa é um patógeno oportunista que tem se destacado quanto à prevalência em casos de infecções hospitalares. Sua ampla resistência aos diversos grupos de antimicrobianos garante a este microrganismo um papel de destaque entre as bactérias mais prevalentes associadas à infecção nosocomial. O objetivo deste estudo foi realizar um levantamento epidemiológico da P. aeruginosa, bem como do seu perfil de susceptibilidade aos antimicrobianos no Hospital das Clínicas da Universidade Federal de Pernambuco. MÉTODOS: Foi realizado um estudo retrospectivo baseado no livro de registro de secreções diversas do laboratório de bacteriologia do Hospital das Clínicas no período 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 laboratório. RESULTADOS: As bactérias mais freqüentes, isoladas das secreções diversas, foram P. aeruginosa (26 por cento) e S. aureus (25 por cento). Quanto à origem, a P. aeruginosa foi isolada principalmente de infecções respiratórias, pois 33 por cento das amostras positivas para esta bactéria foram provinientes de secreções traqueais e 21 por cento nasais. Os antimicrobianos mais eficazes contra a P. aeruginosa foram: amicacina, imipenem, meropenem e aztreonam. CONCLUSÕES: Estes resultados mostram uma alta prevalência de P. aeruginosa, no Hospital das Clínicas da Universidade Federal de Pernambuco. Apesar de apresentar grande resistência a antimicrobianos mais antigos como as cefalosporinas de primeira e segunda geração, assim como cloranfenicol, em geral, este patógeno demonstrou boa sensibilidade às drogas utilizadas na rotina deste hospital.

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 percent) and S. aureus (25 percent). P. aeruginosa was mainly isolated from respiratory infections, with 33 percent of positive samples for this organism from tracheal secretions and 21 percent 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.

Portadores assintomáticos de infecções por Streptococcus pyogenes em duas escolas públicas na cidade do Recife, Pernambuco / Asymptomatic carriers of Streptococcus pyogenes infections in two public schools in Recife, Pernambuco

Objetivos: investigar a prevalência Streptococus pyogenes em secreção de orofaringe de escolares procedentes de duas escolas públicas da cidade Recife. Métodos: estudo epidemiológico e clínico-microbiológico descritivo no qual foram examinados 753 escolares. A cultura bacteriana da secreção orofaringeana foi realizada em ágar-sangue de carneiro 5 por cento e as cepas SBGA identificadas através dos testes de bacitracina, Pyr e aglutinação em látex. Resultados: a faixa etária dos 753 escolares analisados variou de cinco a 19 anos, sendo 54,3 por cento do sexo masculino e 5,7 por cento do sexo feminino. Seis eram portadores assintomáticos de SBGA e foram submetidos ao tratamento com penicilina. Após o tratamento, realizou-se a dosagem da antiestreptolisina o (ASLO), cujos títulos séricos foram inferiores a 200UT. Conclusões: uma prevalência de SBGA de 0,8 por cento foi estimada em portadores assintomáticos, considerada baixa, quando comparado a outros resultados em estudos semelhantes. Sugere-se a realização de outros estudos para estimar a prevalência de SBGA em crianças com faringite e sua relação com a febre reumática aguda.