Dissemination of NDM-producing bacteria in Southern Brazil.

BACKGROUND: The dissemination of NDM-1 carbapenemases (New Delhi Metallo-ß-lactamase) is a global public health problem, mainly in developing countries. The aim of this study was to characterize the spread of NDM-producing bacteria in the Southern Brazilian states analyzing epidemiological, molecular, and antimicrobial susceptibility aspects. METHODS: A total of 10,684 carbapenem-resistant isolates of Enterobacterales, Pseudomonas spp. and Acinetobacter spp. obtained from several hospitals in eight cities in Southern Brazil were screened, and 486 NDM-producing bacteria were selected. RESULTS: The incidence varied from 0.5 to 77 cases/100.000 habitants. ST11, ST15, ST340 and ST674 were the most common in K. pneumoniae. A total of 5 plasmids were identified in one K. pneumoniae strain: Col440I, Col440II, IncFIA(HI1), IncFIB(K), IncFIB(pQil)/ IncFII(K), and IncR. CONCLUSIONS: The number of patients with NDM-producing bacteria has increased in Southern Brazil, whose gene is present in different plasmids, explaining the expansion of this enzyme.

WHO Critical Priority Escherichia coli as One Health Challenge for a Post-Pandemic Scenario: Genomic Surveillance and Analysis of Current Trends in Brazil.

The dissemination of carbapenem-resistant and third generation cephalosporin-resistant pathogens is a critical issue that is no longer restricted to hospital settings. The rapid spread of critical priority pathogens in Brazil is notably worrying, considering its continental dimension, the diversity of international trade, livestock production, and human travel. We conducted a nationwide genomic investigation under a One Health perspective that included Escherichia coli strains isolated from humans and nonhuman sources, over 45 years (1974-2019). One hundred sixty-seven genomes were analyzed extracting clinically relevant information (i.e., resistome, virulome, mobilome, sequence types [STs], and phylogenomic). The endemic status of extended-spectrum ß-lactamase (ESBL)-positive strains carrying a wide diversity of blaCTX-M variants, and the growing number of colistin-resistant isolates carrying mcr-type genes was associated with the successful expansion of international ST10, ST38, ST115, ST131, ST354, ST410, ST648, ST517, and ST711 clones; phylogenetically related and shared between human and nonhuman hosts, and polluted aquatic environments. Otherwise, carbapenem-resistant ST48, ST90, ST155, ST167, ST224, ST349, ST457, ST648, ST707, ST744, ST774, and ST2509 clones from human host harbored blaKPC-2 and blaNDM-1 genes. A broad resistome to other clinically relevant antibiotics, hazardous heavy metals, disinfectants, and pesticides was further predicted. Wide virulome associated with invasion/adherence, exotoxin and siderophore production was related to phylogroup B2. The convergence of wide resistome and virulome has contributed to the persistence and rapid spread of international high-risk clones of critical priority E. coli at the human-animal-environmental interface, which must be considered a One Health challenge for a post-pandemic scenario. IMPORTANCE A One Health approach for antimicrobial resistance must integrate whole-genome sequencing surveillance data of critical priority pathogens from human, animal and environmental sources to track hot spots and routes of transmission and developing effective prevention and control strategies. As part of the Grand Challenges Explorations: New Approaches to Characterize the Global Burden of Antimicrobial Resistance Program, we present genomic data of WHO critical priority carbapenemase-resistant, ESBL-producing, and/or colistin-resistant Escherichia coli strains isolated from humans and nonhuman sources in Brazil, a country with continental proportions and high levels of antimicrobial resistance. The present study provided evidence of epidemiological and clinical interest, highlighting that the convergence of wide virulome and resistome has contributed to the persistence and rapid spread of international high-risk clones of E. coli at the human-animal-environmental interface, which must be considered a One Health threat that requires coordinated actions to reduce its incidence in humans and nonhuman hosts.

Distribution of genes encoding 16S rRNA methyltransferase in plazomicin-nonsusceptible carbapenemase-producing Enterobacterales in Brazil.

BACKGROUND: The presence of 16S rRNA methyltranferases (16S-RMTases) in carbapenemase-producing Enterobacterales (CPE) is a major concern because it inactivates all clinical use of aminoglycosides, including plazomicin. The aim of this study is to investigate the prevalence of 16S-RMTases in CPE nonsusceptible to plazomicin collected in different Brazilian hospitals. METHODS: All isolates with plazomicin MIC ≥ 4 µg/mL (n = 67) were screened for the presence of 16S-RMTases by sequencing. RESULTS: 54 (80.6%) isolates encoded 16S-RMTase genes (41 rmtB1, 7 armA, 3 rmtD2, 1 rmtD1 and 2 rmtC). Among 41 samples rmtB1 positive, 40 co-harbored blaKPC-2 and 1 blaOXA-48 gene. Of the seven isolates harboring armA gene, 6 were New Delhi Metallo-beta-lactamase (NDM)-producer. rmtD was only found in isolates Klebsiella pneumoniae Carbapenemase (KPC)-producers, one in Serratia marcescens with rmtD2, not reported in Brazil. CONCLUSION: The co-existence of 16S-RMTase and CPE is worrisome because of limited treatment options and the endemic characteristic of (KPC) and NDM in Brazil.

Evaluation of EDTA and Dipicolinic Acid in Broth Microdilution with Polymyxin B as a Phenotypic Test to Detect the mcr-1 Gene.

Polymyxins (colistin and polymyxin B) have recently regained significant importance as last-line drugs to treat infectious diseases due to multidrug-resistant gram-negative bacteria. However, resistance to polymyxins has increased, and the recognition of plasmid-mediated resistance (by the mcr gene) has led to an epidemiological concern. We aimed to evaluate the reduction of the polymyxin B minimum inhibitory concentration (MIC) in the presence of EDTA or dipicolinic acid (DPA) by using the broth microdilution (BMD) method for phenotypic screening of acquired polymyxin resistance mediated by the mcr-1 gene. Overall, 94 Enterobacterales (48 polymyxin-resistant and 46 polymyxin-susceptible) were evaluated: 47 mcr-1 positive (36 Escherichia coli, 2 Klebsiella pneumoniae, and 9 Salmonella spp.) and 47 mcr-1 negative (3 E. coli and 44 K. pneumoniae-27 isolates with MIC from ≤0.125 to 8 µg/mL and 20 isolates with MIC from 16 to 64 µg/mL). Results were categorized as positive when the chelator decreased the original BMD MIC by ≥2 logs. The majority (95.7%) of mcr-1 positive isolates displayed at least a 3 log dilution decrease in the MIC of polymyxin B with EDTA or DPA. The EDTA-based BMD assay detected 45 mcr-1-positive isolates, with only one false-positive among the mcr-1-negative isolates (sensitivity [SN], 95.7%; specificity [SP], 97.9%), whereas the DPA-based BMD assay detected 44 mcr-1-positive isolates (SN, 93.6%; SP, 95.7%), with two false-positive results. The accuracy of EDTA- and DPA-based BMD assays were 97% and 95%, respectively. The EDTA- and DPA-based assays were demonstrated to be reliable methods to detect mcr-1 positive isolates with excellent accuracy.

Comparison of automated and conventional microbiological examination of donated human cardiac tissue in heart valve banking.

Most tissue banks use the conventional method; however, the automated method has advantages over the conventional method. The aim of this study was to compare the conventional and automated methods of culture in human cardiac tissue using an artificial contamination model. Myocardial samples were contaminated with sequential concentration (104 to 10-1 CFU/mL) with Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Candida albicans. Cultures were obtained from solution were the fragment was immersed and minced tissue, before and after the routine decellularization solution, with automated and conventional culture methods. Automated and conventional methods were compared and a p value ≤ 0.05 was considered significant. Staphylococcus aureus presented a significantly higher growth in the automated method, as well as faster than the conventional (p < 0.05). The positivity for growth in the automated method was higher in concentrated inoculum (> 102 CFU/mL) (p < 0.05). The growth in the automated method was significantly faster than conventional when inoculum concentration was above 103 CFU/mL. The automated culture method is faster than conventional method with a higher positivity in a contaminated model of myocardial and transport solution used in tissue banks.

Phenotypic and molecular characterization of 942 carbapenem-resistant Enterobacteriaceae (CRE) in southern Brazil.

Carbapenem-resistant Enterobacteriaceae (CRE) is a major international health problem, and its identification in developing countries is based exclusively on phenotypic methods. The aim of this study was to assess the sensitivity and related parameters of the modified Hodge test (MHT). The assessment was performed in a large number of isolates obtained from different hospitals in several cities of a south Brazilian state. Bacterial species were identified using an automated method. The MHT was performed according to the guidelines set by the CLSI. The gene blaKPC was amplified in order to confirmation CRE expression. The sensitivity, specificity, positive, and negative predictive values were calculated. A total of 942 isolates were submitted to the reference laboratory for confirmation; 143 showed a negative MHT (15.18%) result, while 784 were positive (83.23%), and 15 samples displayed an indeterminate MHT (1.59%) result. All samples expressed the KPC-2 enzyme. Sensitivity, specificity, positive, and negative predictive percentiles were 99%, 89%, 98%, and 99% respectively. We conclude that the modified Hodge test is a reliable test for the prediction of KPC-producing bacteria.

Molecular epidemiology of Klebsiella pneumoniae carbapenemase-producing Enterobacteriaceae in different facilities in Southern Brazil.

BACKGROUND: Klebsiella pneumoniae carbapenemase-producing K pneumoniae (KPC-KP) outbreaks have been reported in many countries, including Brazil. The incidence of KPC-KP infection has increased in the first semester of 2011 in Curitiba, the capital of Parana, in Southern Brazil.The aim of this study was to characterize the infections and clonal diversity of KPC-KP isolates from several institutions in Curitiba. METHODS: KPC-KP from several clinical samples and rectal swabs taken between April 2010 and July 2012 were included. One isolate per patient was evaluated. All isolates were submitted to polymerase chain reaction (PCR) for blaKPC. The genetic relatedness was evaluated using strain clustering by an automated repetitive extragenic palindromic (rep) PCR-based typing system. RESULTS: There were 641 samples that were positive for K pneumoniae carbapenemase-2 carbapenemase. There were 129 samples randomly selected for clonality evaluation. PCR and strain clustering by the automated rep PCR-based typing system identified 7 clones (A-C and E-H). Clone E was identified in only 1 hospital, and all other clones were found in >2 hospitals. Clones C and G were the most disseminated among hospitals. The infection and colonization occurred in 14 out of the 32 main hospitals in town. Similar clones were found in 2 hospitals that are administered by the same group. Another clone (H) was found in 2 hospitals receiving patients from the same municipal emergency unit. CONCLUSION: The KPC-KP outbreak in Curitiba is polyclonal, and the source is unknown. Some hospitals share the same clones.

MALDI-TOF: a useful tool for laboratory identification of uncommon glucose non-fermenting Gram-negative bacteria associated with cystic fibrosis.

The predisposition of patients with cystic fibrosis (CF) for recurrent pulmonary infections can result in poor prognosis of the disease. Although the clinical significance in CF of micro-organisms, such as Staphylococcus aureus, Haemophilus influenzae and Pseudomonas aeruginosa, is well established, the implication of uncommon glucose non-fermenting Gram-negative bacilli (UGNF-GNB) in respiratory samples from CF patients is still unclear. Because of limitations of traditional methods used in most clinical laboratories, the accurate identification of these microbes is a challenge. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) is an alternative tool for efficient identification of bacteria. This was a retrospective study to evaluate different identification methods in a collection of UGNF-GNB isolated from children with CF during a period of three years. The performance of MALDI-TOF was compared to that of 16S rDNA gene sequencing and to a conventional and automated phenotypic identification. The discriminatory power of MALDI-TOF (75.0 % agreement) was superior to automated techniques (67.1 % agreement) and to conventional phenotypical identification (50.0 % agreement). MALDI-TOF also demonstrated high accuracy in identifying Stenotrophomonas maltophilia, Achromobacter xylosoxidans and Chryseobacterium indologenes, but had limited utility in identifying Pandoraea spp. and some species of Acinetobacter and Chryseobacterium (other than C. indologenes). Although MALDI-TOF identified only 75 % of the isolates in comparison with 16S rDNA gene sequencing, the prompt identification and high discriminatory power exhibited by MALDI-TOF make it a useful tool for the characterization of micro-organisms that are difficult to identify using routine methods.

Effects of low intensity laser in in vitro bacterial culture and in vivo infected wounds.

OBJECTIVE: to compare the effects of low intensity laser therapy on in vitro bacterial growth and in vivo in infected wounds, and to analyze the effectiveness of the AsGa Laser technology in in vivo wound infections. METHODS: in vitro: Staphylococcus aureus were incubated on blood agar plates, half of them being irradiated with 904 nm wavelength laser and dose of 3J/cm² daily for seven days. In vivo: 32 male Wistar rats were divided into control group (uninfected) and Experimental Group (Infected). Half of the animals had their wounds irradiated. RESULTS: in vitro: there was no statistically significant variation between the experimental groups as for the source plates and the derived ones (p>0.05). In vivo: there was a significant increase in the deposition of type I and III collagen in the wounds of the infected and irradiated animals when assessed on the fourth day of the experiment (p=0.034). CONCLUSION: low-intensity Laser Therapy applied with a wavelength of 904 nm and dose 3J/cm² did not alter the in vitro growth of S. aureus in experimental groups; in vivo, however, it showed significant increase in the deposition of type I and III collagen in the wound of infected and irradiated animals on the fourth day of the experiment.

Effects of low intensity laser in in vitro bacterial culture and in vivo infected wounds / Efeitos do laser de baixa intensidade em cultura bacteriana in vitro e ferida infectada in vivo

OBJECTIVE: to compare the effects of low intensity laser therapy on in vitro bacterial growth and in vivo in infected wounds, and to analyze the effectiveness of the AsGa Laser technology in in vivo wound infections. METHODS: in vitro: Staphylococcus aureus were incubated on blood agar plates, half of them being irradiated with 904 nm wavelength laser and dose of 3J/cm2 daily for seven days. In vivo: 32 male Wistar rats were divided into control group (uninfected) and Experimental Group (Infected). Half of the animals had their wounds irradiated. RESULTS: in vitro: there was no statistically significant variation between the experimental groups as for the source plates and the derived ones (p>0.05). In vivo: there was a significant increase in the deposition of type I and III collagen in the wounds of the infected and irradiated animals when assessed on the fourth day of the experiment (p=0.034). CONCLUSION: low-intensity Laser Therapy applied with a wavelength of 904nm and dose 3J/cm2 did not alter the in vitro growth of S. aureus in experimental groups; in vivo, however, it showed significant increase in the deposition of type I and III collagen in the wound of infected and irradiated animals on the fourth day of the experiment. .

OBJETIVO: comparar os efeitos da terapia a laser de baixa intensidade no crescimento bacteriano in vitro e em feridas infectadas in vivo, e analisar a efetividade da tecnologia Laser AsGa, em feridas infectadas in vivo. MÉTODOS: in vitro: cepas de Staphylococcus aureus foram incubadas em placas de agar-sangue e irradiadas com laser de 904nm de comprimento de onda e dose de 3J/cm2, diariamente durante sete dias. In vivo: 32 ratos machos Wistar foram distribuídos em Grupo Controle (Não Infectado) e Grupo Experimental (Infectados). Metade dos ratos tiveram suas feridas irradiadas e a outra metade não irradiada, como realizado no estudo in vitro. RESULTADOS: in vitro: não houve variação estatística significativa entre os grupos experimentais, considerando as placas matrizes e derivadas (p>0,05). In vivo: houve aumento significativo na deposição de colágeno tipo I e III na cicatriz do grupo dos animais infectados e irradiados, quando avaliados no quarto dia de experimento (p=0,034). CONCLUSÃO: a Low-Intensity Laser Therapy aplicada com comprimento de onda de 904nm e dose de 3J/cm2, in vitro: não alterou o crescimento de S. aureus nos grupos experimentais. In vivo: mostrou aumento significativo na deposição de colágeno tipo I e III na cicatriz no grupo dos animais infectados e irradiados no quarto dia de experimento. .

Molecular epidemiology characterization of OXA-23 carbapenemase-producing Acinetobacter baumannii isolated from 8 Brazilian hospitals using repetitive sequence-based PCR.

The typing of multidrug-resistant Acinetobacter baumannii isolates is important for the control and prevention of hospital outbreaks. This study aimed to analyze the molecular epidemiology of 46 OXA-23 carbapenemase-producing A. baumannii strains and compare them to previously described local and international clones (ICs). Isolates were recovered during May 2009-August 2011, from 8 different hospitals in the state of Parana (Brazil). The molecular profiles were determined by repetitive extragenic palindromic PCR. Seven different clusters were identified (A to G). Thirty-two isolates were clustered in the same pattern (clone A), which belong to IC 4.

Perfil epidemiológico de casos de surtos de doenças transmitidas poralimentos ocorridos no Paraná, Brasil / Epidemiological profile of foodborne diseases outbreaks occurred in Parana state, Brazil

A investigação dos surtos de Doenças Transmitidas por Alimentos (DTA) é desafiadora. Para a Vigilância em Saúde é fundamental a identificação de grupos e fatores de risco associados às DTA. O objetivo deste trabalho foi caracterizar o perfil epidemiológico dos surtos de DTA ocorridos nos 29 municípiosque compõe a 2ª Regional de Saúde, da Secretaria Estadual de Saúde do Paraná, Brasil, entre 2005-2008. Tabularam-se 472 laudos de alimentos do Laboratório Central do Estado do Paraná (LACEN-PR) e fichas notificadas no Sistema de Informação de Agravos de Notificação (SINAN) de casos suspeitos de DTA. O grupo mais acometido foi de indivíduos do sexo feminino com 50,51% dos casos. Indivíduos entre 20 a 49 anos de idade foram os mais frequentemente envolvidos com 49,50% das ocorrências. Restaurantes (27,65%) e residências dos indivíduos acometidos (36,17%) foram os locais de preparo econsumo mais frequentemente envolvidos. Manipulação/preparo inadequados dos alimentos (36,95%) eforma de consumo inadequada (23,90%) foram os fatores causais mais predominantes. A maior parte dasconfirmações dos surtos fundamentou-se em achados clínico-epidemiológicos (44,68%). Com relaçãoaos micro-organismos identificados nas amostras analisadas, os resultados apontaram predominância Escherichia coli (indicador sanitário), seguido pelo Bacillus cereus e Staphyloccus aureus. Os alimentos mais frequentemente envolvidos foram o leite (29,68%) e as frutas, verduras e legumes (24,21%). Com a pesquisa é possível orientar medidas preventivas e de controle de DTA na população pesquisada.

The investigation of outbreaks of Food Borne Diseases (FBD) is challenging. For the Health Surveillance is essential to identify groups and risk factors associated with the DTA. The aim of this study was to characterize the epidemiology of outbreaks of DTA occurred in 29 counties that comprise the 2nd Regional Health, the Department of Health of Paraná, Brazil, from 2005-2008. Tabulated to 472 reports of food the Central Laboratory of the State of Paraná (LACEN-PR) chips and notified the Information System for Notifiable Diseases (SINAN) of suspected cases of DTA. The group most affected were females with 50.51% of the cases. Individuals between 20 and 49 years of age were most often involved in 49.50% of cases. Restaurants (27.65%) of affected individuals and households (36.17%) were the preparation and consumption sites most frequently involved. Manipulation / inadequate preparation of food (36.95%) and inadequate form of consumption (23.90%) were the most prevalent causative factors.Most confirmations of outbreaks was based on clinical and epidemiological (44.68%). With respect to micro-organisms identified in the samples analyzed, the results showed predominance Escherichia coli(health indicator), followed by Bacillus cereus and Staphylococcus aureus. The foods most frequently involved were milk (29.68%) and fruits and vegetables (24.21%). With this study can guide preventive measures and control of DTA in the population studied.

Validação do teste de inibição pelo ácido aminofenilborônico para triagem de Klebsiella pneumoniae carbapenemases (KPC) / Validation of inhibition test by aminophenyl boronic acid to Klebsiella pneumoniae carbapenemases (KPC) screening

INTRODUÇÃO: A produção de enzimas Klebsiella pneumoniae carbapenemase (KPC) tem se tornado um importante e preocupante mecanismo de resistência, e ensaios que combinem alta sensibilidade e alta especificidade para a detecção dessas enzimas são escassos. OBJETIVO: Validar o teste de inibição pelo ácido 3-aminofenilborônico como método de triagem fenotípica de cepas produtoras de enzima KPC, comparando os resultados obtidos com os de testes confirmatórios por reação em cadeia da polimerase (PCR). METODOLOGIA: Avaliou-se o uso do ácido 3-aminofenilborônico impregnado em discos de antibióticos de imipenem, meropenem e ertapenem. Foram testadas 36 cepas positivas e 12 negativas, todas confirmadas por PCR. Foram ainda testadas três concentrações diferentes de ácido borônico: 300, 400 e 600 µg. RESULTADOS: Entre as cepas positivas testadas, o resultado mais adequado se deu com a adição do composto em disco contendo ertapenem, com especificidade de 100%, porém com sensibilidade de apenas 50%. CONCLUSÃO: Novos estudos são necessários, sobretudo no que diz respeito à padronização da técnica e aos insumos utilizados, pois o método se revela promissor na triagem de cepas produtoras de KPC.

INTRODUCTION: The production of Klebsiella pneumoniae carbapenemases enzymes (KPC) has become an important and worrisome resistance mechanism. Furthermore, tests that combine high sensitivity and high specificity for the detection of these enzymes are scarce. OBJECTIVE: To validate the inhibition test by 3-aminophenyl boronic acid as a phenotypic screening method for KPC-producing strains by comparing the results with confirmatory polymerase chain reaction testing (PCR). METHODS: We evaluated the use of 3-aminophenyl boronic acid applied on disks with imipenem, meropenem and ertapenem antibiotics. 36 strains were positive and 12 were negative, all confirmed by PCR. Three different concentrations of boronic acid were also tested: 300, 400 and 600 µg. RESULTS: Among the positive strains, the results were more accurate with the addition of the compound to the ertapenem disk, presenting 100 % specificity and 50% sensitivity. CONCLUSION: Further studies are required, mainly regarding the standardization of the technique and materials, since the method seems to be promising as to the screening of KPC strains.

Risk factors for KPC-producing Klebsiella pneumoniae bacteremia

The molecular epidemiology of carbapenemase-producing Klebsiella pneumoniae (KPC) has been largely investigated, but limited clinical information is available. A case-control study was performed to evaluate the risk factors for KPC bacteremia in hospitalized patients. Cases were patients with KPC bacteremia and controls were patients with non-KPC bacteremia. A total of 85 patients were included, 18 (21.2%) were KPC, and 67 (78.8%) were non-KPC (40 [59.7%] of them were extended-spectrum beta-lactamase producers). All KPC isolates were type 2 producers. These isolates belong to five distinct clones. Multivariate analysis showed that age (odds ratio [OR], 1.06; 95% confidence interval [CI], 1.02 - 1.11; p = 0.004), presence of mechanical ventilation (OR, 11.1; 95% CI, 1.92 - 63.3; p = 0.007) and fluoroquinolone exposure during hospitalization (OR, 28.9; 95% CI, 1.85 - 454.6; p = 0.02) were independent risk factors for KPC in patients with K. pneumoniae bacteremia. Factors associated with severity of illness, such as age and mechanical ventilation, seem to be the main risks factors for KPC. Fluoroquinolones use might be a risk factor for KPC bacteremia. Further investigations on risk factors for KPC are warranted.

Risk factors for KPC-producing Klebsiella pneumoniae bacteremia.

The molecular epidemiology of carbapenemase-producing Klebsiella pneumoniae (KPC) has been largely investigated, but limited clinical information is available. A case-control study was performed to evaluate the risk factors for KPC bacteremia in hospitalized patients. Cases were patients with KPC bacteremia and controls were patients with non-KPC bacteremia. A total of 85 patients were included, 18 (21.2%) were KPC, and 67 (78.8%) were non-KPC (40 [59.7%] of them were extended-spectrum beta-lactamase producers). All KPC isolates were type 2 producers. These isolates belong to five distinct clones. Multivariate analysis showed that age (odds ratio [OR], 1.06; 95% confidence interval [CI], 1.02 - 1.11; p = 0.004), presence of mechanical ventilation (OR, 11.1; 95% CI, 1.92 - 63.3; p = 0.007) and fluoroquinolone exposure during hospitalization (OR, 28.9; 95% CI, 1.85 - 454.6; p = 0.02) were independent risk factors for KPC in patients with K. pneumoniae bacteremia. Factors associated with severity of illness, such as age and mechanical ventilation, seem to be the main risks factors for KPC. Fluoroquinolones use might be a risk factor for KPC bacteremia. Further investigations on risk factors for KPC are warranted.

Rastreabilidade das causas de não conformidades em meios de cultura / Traceability of the causes of non-compliance in culture media

O processo de produção de meios de cultura deve garantir a liberação do produto dentro das especificações requeridas em normas e legislações. Quando o produto apresentar resultado não conforme com as especificações, deve-se buscar a causa raiz do problema e implantar ações corretivas. A rastreabilidade das causas de não conformidades deve ser uma etapa de decisão rápida. Objetivando elaborar ferramentas da qualidade voltadas a esta rastreabilidade e verificar a sua aplicabilidade em estudos de casos,foram analisados manuais e registros de produção e controle dos produtos. Foi aplicada a metodologia de rainstorming, a produção e o controle foram mapeados e com o Diagrama de Causa e Efeito foi possível estabelecer os pontos críticos. As ferramentas da qualidade propostas foram Fluxogramas e Folhas de Verificação de Processos, cuja aplicabilidade foi avaliada em lotes de meios de cultura não conformes. Os resultados mostraram que as ferramentas foram capazes de indicar, localizar e confirmar as nãoconformidades e as suas origens. Porém, para a investigação aprofundada da causa torna-se necessário, além destas ferramentas, o conhecimento técnico em microbiologia, a pesquisa em literaturas e Brainstorming permanente, possibilitando tomada de decisãoconfiável em desvios da qualidade dos meios de cultura.

The process of production of culture media should ensure the release of the product within the specifications required by rules and laws. When the product does not produce results consistent with the specifications, you should seek the root cause of the problem and implement corrective actions. The traceability of the causes of non-conformities should be a stage of rapid decision. Aiming at producing quality tools focussed on traceability and verifying its applicability in case studies, manuals and records were analyzed for the production and control of products. The methodology of Brainstorming was used, the productionand control have been mapped and, with the Diagram of Cause and Effect, it was possible to establish the critical points. The quality tools proposed were flow charts and Verification Process sheets, whose applicability was evaluated in batches of noncompliant culture media. The results showed that the tools were able to indicate, locate and confirm the non-conformities and their origins. However, for a detailed case nvestigation, besides these tools, technical knowledge in microbiology, research in different literature and permanent Brainstorming, are necessary thus enabling reliable decision making related to gaps in the quality of culture media.

Detecção de metalo-beta-lactamases em amostras de pseudomonas aeruginosa isoladas em Porto Alegre, Rio Grande do Sul / Detection of metallo-beta-lactamase producing pseudomonas aeruginosa isolated in Porto Alegre,Rio Grande do Sul, Brazil

As metalo-beta-lactamases (MBLs) pertencem ao grupo 3 de beta-lactamases de espectro estendido, fazendo parte de uma classefuncional de metaloenzimas classificadas com base na sua capacidade de hidrolisar os carbapenemicos como o Imipenem e em suas caracteristicas de serem inibidas por agentes quelantes como o acido etilenodiamino tetra-acetico (EDTA) e acido 2-mercaptopropionico (2-MPA). Este estudo objetivou a deteccao de MBLs em amostras nosocomiais de P. aeruginosa resistentes a Ceftazidima, provenientes de um hospital de medio porte de Porto Alegre-RS. Foram analisados 113 isolados clinicos atraves de testes fenotipicos, sendo: 80 (70,7%) do trato respiratorio inferior, 24 (21,2%) urina, 3 (2,7%) hemocultura, 3 (2,7%) ponta de cateter, 2 (1,8%) colecao de cavidadese 1 (0,9%) liquor. Com relacao a producao de MBLs, detectou-se 35 amostras (31,0%) como sendo produtoras.

Metallo-beta-lactamases belongs to the group 3 of extended spectrum beta-lactamases, making part of a common functional class of metal enzymes classified based in their ability to hydrolyze Imipenem and in your characteristics of been inhibited by ions of chelating metals, like EDTA and 2-MPA. The aim of this study was detect MBLs in samples of P. aeruginosa resistant to ceftazidime in the patients of PortoAlegre- RS Brazil. The samples was provided of different clinical specimens like: 80 (70,7%) respiratory tract, 24 (21,2%) urine, blood 3 (2,7%), catheter 3 (2,7%), 2 (1,8%) corporeal secretions and 1 (0,9%) liquor. With regard to the MBLs production, we detect 35 (31%) as being producing.

An outbreak of Streptococcus dysgalactiae subsp equisimilis in a hospital in the south of Brazil

The beta-hemolytic group C streptococci (Lancefield's group) has been considered an emergent human pathogen, showing an important role as an opportunist agent, being responsible for nosocomial infections and outbreaks. This study is reporting the first outbreak of nosocomial infection caused by Streptococcus dysgalactiae subsp. equisimilis in Brazil. From January, 2002, to December, 2004, S. equisimilis was isolated in 67/207 (32.37 percent) samples from secretions of patients' infected wounds, interned at the Hospital of Sanitary Dermatology in the State of Paraná (HDSPR). The prevalence of this microorganism increased from 11/42 (26.19 percent) in 2002, 14/65 (21.54 percent) in 2003 to 42/100 (42.00 percent) in 2004. This increase was statistically significant (p=0.024), and this microorganism became the most frequently isolated in these patients, overtaking the rates of isolation of Pseudomonas aeruginosa. The S. equisimilis grew in pure culture, as a unique microorganism, in six samples (2.9 percent) out of 207. Fresh feces of 15 animals (horses and sheep) living in the proximities of the hospital were also examined and three of them positive for S. equisimilis. The biochemical profile of the strains isolated from the patients and from the animals was the same. These animals might have been the source of the dissemination of the outbreak in the hospital. New studies will be necessary to confirm the genetic relationship between the strains isolated from patients and animals.

O estreptococo beta-hemolítico do grupo C de Lancefield tem sido considerado patógeno humano emergente, mostrando importante papel como agente oportunista, implicado algumas vezes em infecções hospitalares e surtos. Este estudo está relatando o primeiro surto de infecção hospitalar causado pelos Streptococcus dysgalactiae subsp. equisimilis no Brasil. De janeiro de 2002 a dezembro de 2004, isolou-se S. equisimilis em 67/207 (32,37 por cento) das amostras de secreções de lesões de feridas coletadas de pacientes internados no Hospital de Dermatologia Sanitária do Paraná (HDSPR). A prevalência deste microrganismo aumentou de 11/42 (26,19 por cento) em 2002, 14/65 (21,54 por cento) em 2003 para 42/100 (42,00 por cento) em 2004. Este aumento foi estatisticamente significante (p=0.024), tornando este microrganismo o mais freqüentemente isolado nos pacientes, ultrapassando as taxas de isolamento de Pseudomonas aeruginosa. Em seis amostras (2,9 por cento) entre as 207 examinadas, S. equisimilis cresceu em cultivo puro, como único microrganismo. Também foram examinadas fezes frescas de 15 animais (cavalos e ovelhas) que vivem nas proximidades do hospital, e três amostras foram positivas para S. equisimilis. O perfil bioquímico encontrado entre os isolados dos pacientes foi o mesmo encontrado entre os isolados das fezes dos animais. Acredita-se que estes animais possam ter sido a fonte de disseminação do surto no hospital. Novos estudos serão necessários para confirmar o relacionamento genético entre os isolados dos pacientes e animais.