Maria Amélia Vieira Maciel

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.

Pseudomonas aeruginosa: freqüência de resistência a múltiplos fármacos e resistência cruzada entre antimicrobianos no Recife/PE

BACKGROUND AND OBJECTIVES: The frequency of multiple-antibiotic resistant bacteria has been increasing in recent years. Among the gram-negative bacteria Pseudomonas aeruginosa (P. aeruginosa) shows a great propensity for the development of multidrug resistance mechanisms. The objective of this study was to identify the profile of susceptibility to antibiotics, the frequency of multidrug resistance and the cross-resistance between drugs of P. aeruginosa strains in two tertiary hospitals in Recife, Pernambuco. METHODS: The study was carried out between September 2004 and January 2006. The antimicrobial susceptibility testing was performed in 304 strains of P. aeruginosa by the disc diffusion method in accordance with National Committee for Clinical and Laboratory Standards (NCCLS) guidelines. RESULTS: The most frequent materials were urine (26.7%) and respiratory tract secretion (26.1%) The antibiotics tested and their respective susceptibilities were as follows: piperacillin-tazobactam (66.2%); aztreonam (59.8%); amikacin (59.4%); meropenem (58.2%); imipenem (57.7%); ciprofloxacin (49.7%); gentamicin and cefepime (48.6%); ceftazidime (30%) and cefotaxime (6.8%). A high prevalence of multi-resistance was detected. Half (49.7%) the strains showed resistance to three or more antibiotics and 28% were resistant to six antimicrobials or more. Also, cross-resistance between the beta-lactams (carbapenems and piperacilin/tazobactam) and aminoglicosides and quinolones was between 22.9% and 38.1%. These drugs are commonly combined in the treatment of severe infections caused by Pseudomonas, which reflects the difficulty in choosing the appropriate option for combination therapy. CONCLUSIONS: The frequency of multidrug-resistant strains of P. aeruginosa in this study was similar to other hospitals in Brazil and higher than in other countries. In order to reduce the frequency of these multiresistant clones, epidemiologic surveillance and the rational use of antibiotic protocols need to be urgently implemented.

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.

Prevalence and risk factors for Staphylococcus aureus in health care workers at a University Hospital of Recife-PE

Staphylococcus aureus is the main human pathogen that colonizes individuals in general population. The objective of the study was evaluate the epidemiological and sensitivity profile of S. aureus lineage, isolated in health care workers (HCW) of a University Hospital in Pernambuco state, Brazil. Biological samples of hands and nasal cavities were sown in agar sheep blood. Colonies under suspicion of being S. aureus were identified using Gram staining, catalase test and coagulase, mannitol-salty agar fermentation and DNAse agar. The resistance to mupirocin was analyzed through the Kirby Bauer technique. In relation to methicillin and vancomycin the determination was by the minimum inhibitory concentration method (E-test). From the 202 HCW evaluated, 52 were colonized by S. aureus (25,7%). The factors associated to the colonization by S. aureus were: age-group, professional category, use of individual protection equipments (frequency and numbers). All S. aureus isolate lineages were sensitive to mupirocin and vancomycin, and three of them were identified as methicillin-resistant. The prevalence of MSSA and MRSA among HCW was considered low and was below the results described in the literature. The isolate S. aureus lineages have shown low resistance profile.

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.

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

INTRODUCTION Methicillin-resistant Staphylococcus aureus (MRSA) strains have been responsible for many nosocomial outbreaks. Within hospitals, colonized employees often act as reservoirs for the spread of this organism. This study collected clinical samples of 91 patients admitted to the intensive care unit (ICU), hemodialysis/nephrology service and surgical clinic, and biological samples from the nasal cavities of 120 professionals working in those environments, of a University Hospital in Recife, in the State of Pernambuco, Brazil. The main objective of this study was to determine the occurrence and dissemination of methicillin- and vancomycin-resistant Staphylococcus spp. METHODS The isolates obtained were tested for susceptibility to oxacillin and vancomycin and detection of the mecA gene. In addition, the isolates were evaluated for the presence of clones by ribotyping-polymerase chain reaction (PCR). RESULTS MRSA occurrence, as detected by the presence of the mecA gene, was more prevalent among nursing technicians; 48.1% (13/27) and 40.7% (11/27) of the isolates were from health professionals of the surgical clinic. In patients, the most frequent occurrence of mecA-positive isolates was among the samples from catheter tips (33.3%; 3/9), obtained mostly from the hemodialysis/nephrology service. Eight vancomycin-resistant strains were found among the MRSA isolates through vancomycin screening. Based on the amplification patterns, 17 ribotypes were identified, with some distributed between patients and professionals. CONCLUSIONS Despite the great diversity of clones, which makes it difficult to trace the source of the infection, knowledge of the molecular and phenotypic profiles of Staphylococcus samples can contribute towards guiding therapeutic approaches in the treatment and control of nosocomial infections.