Detection of microorganisms with antibacterial activities from different industrial wastes and GC-MS analysis of crude microbial extract / Detecção de microrganismos com atividades antibacterianas de diferentes resíduos industriais e análise por GC-MS do extrato microbiano bruto

Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence [...].

Muitos microrganismos do solo, ou seja, bactérias e fungos produzem metabólitos secundários chamados antibióticos. Eles são usados para tratamento de algumas doenças bacterianas, fúngicas e protozoárias em humanos. Há necessidade de isolamento de um amplo espectro de antibióticos de microrganismos devido ao surgimento de resistência aos antibióticos. No presente estudo, duas bactérias produtoras de antibióticos, Klebsiella pneumoniae e Bacillus cereus, foram isoladas da indústria farmacêutica e de ração avícola de Hattar, Haripur, Paquistão. Um total de 10 amostras de resíduos foi coletado de diferentes indústrias (mármore, ghee, sabão, mineral, aço, ração para aves, farmacêutica, Qarshi, cosmética e vidro). Trinta e três cepas bacterianas foram isoladas de resíduos industriais dessas dez diferentes indústrias. Quatorze das 33 cepas bacterianas exibiram atividades antimicrobianas contra pelo menos um dos micróbios de teste considerados neste estudo, incluindo Escherchia coli, Staphylococcus aureus e Salmonella typhi. As bactérias foram isoladas pela técnica de placa de diluição em série padrão. A caracterização morfológica dos isolados foi feita por coloração de gram. Nove isolados bacterianos de 14 foram inicialmente identificados como B. cereus e cinco como K. pneumoniae por meio de caracterização bioquímica. As atividades antibacterianas foram testadas pelo método de difusão em poço. O número máximo de bactérias produtoras de antibióticos foi isolado da indústria farmacêutica e de ração avícola com base nos resultados da triagem primária, os isolados mais potenciais S9, S19, S20, S22 e S23 foram selecionados para a triagem secundária. A atividade máxima contra E. coli e S. aureus foi registrada pelo isolado bacteriano S19, ou seja, zonas de inibição de 6,5 mm e 9 mm, enquanto S20 mostrou zonas de 7,5 mm e 6 mm, respectivamente. A identificação molecular foi realizada com base na análise da sequência 16S [...].

Bacteria isolated from the lower respiratory tract of sheep and their relationship to clinical signs of sheep respiratory disease / Bactérias detectadas no trato respiratório inferior de ovinos e a relação com as manifestações clínicas da doença respiratória ovina

Respiratory diseases are among the most important diseases in sheep flocks. Herein was studied the bacterial etiology of respiratory disease and the clinical signs of 99 female and male sheep breed in the states of São Paulo (SP) and Rio de Janeiro (RJ), Brazil. After physical examination of animals, tracheobronchial flushing samples were obtained. The usual bacteria and Mycoplasma spp. were searched, as well as their association with the clinical status and clinical signs of sheep with respiratory disease. The main observed signs were: tachypnea (75%), increase of rectal temperature (09.4%), mucopurulent/purulent nasal discharge (21.9%), cough (25%), dyspnea (31.2%), changes of lung sounds at auscultation (87.5%) and chest percussion (28.1%) in pneumonic sheep. Non-fermenting gram-negative bacteria and Bacillus sp. were the most isolated bacteria. Microorganisms of the Mollicutes class were molecularly (PCR) detected in 33.3% of the animals. In addition, the specific detection of M. mycoides subsp. capri was described for the first time in sheep from the state of São Paulo, Brazil.(AU)

A doença respiratória é uma das doenças mais importantes em rebanhos ovinos. Esta pesquisa teve como objetivo determinar a etiologia bacteriana da doença respiratória e sua relação com sinais clínicos em ovinos criados nos estados de São Paulo e Rio de Janeiro, Brasil. Noventa e nove ovelhas machos e fêmeas dos Estados de São Paulo (SP) e Rio de Janeiro (RJ) foram estudadas. Após o exame físico, amostras de lavagem traqueobrônquica foram obtidas. A presença de bactérias aeróbias e Mycoplasmaspp. foram estudados, assim como a associação entre os microrganismos e estado clínico e sinais clínicos de doença respiratória em ovinos. As principais manifestações clínicas observadas foram: taquipneia (75%), alta temperatura retal (09,4%), secreção nasal mucopurulenta/purulenta (21,9%), tosse (25%), dispneia (31,2%), sons pulmonares alterados na ausculta (87,5%) e na percussão torácica (28,1%) em ovelhas pneumônicas. Bactérias gram-negativas não fermentadoras e Bacillus sp. foram as bactérias mais isoladas. Microrganismos da classe Mollicutes foram detectados molecularmente (PCR) em 33,3% dos ovinos. Além disso, descreve-se pela primeira vez no estado de São Paulo, Brasil, a detecção do M. mycoides subsp. capri na espécie ovina utilizando a reação de polimerase em cadeia.(AU)

Reutilização do detergente enzimático: avaliação do impacto da contaminação microbiana da solução na efetividade da limpeza de aparelhos endoscópicos gastrointestinais / Enzymatic detergent reuse: evaluation of the impact of the solution's microbial contamination on the cleaning effectiveness of gastrointestinal endoscopes

No Brasil, é recomendado que durante a limpeza dos Produtos para Saúde (PPS) o detergente utilizado possua ação enzimática. Embora a Resolução da Diretoria Colegiada nº 55 de 14 de novembro de 2012 da Agência Nacional de Vigilância Sanitária desaconselhe a reutilização desta solução de limpeza, sabe-se que na prática clínica elas são reaproveitadas por diversas vezes para imersão de PPS, como os aparelhos endoscópicos, o que pode comprometer a efetividade da ação do detergente enzimático e com isso a segurança no processamento do PPS. Esta pesquisa objetivou avaliar a carga microbiana presente na solução de detergente enzimático durante sua reutilização na limpeza manual de aparelhos endoscópicos gastrointestinais. Tratou-se de um estudo transversal realizado em um serviço de endoscopia digestiva de um hospital universitário de Belo Horizonte e no Laboratório de Microbiologia Oral e Anaeróbios do ICB/UFMG. A amostra foi composta por 57 aparelhos endoscópicos e 76 alíquotas de solução de detergente enzimáticos coletadas de diversos reusos de 19 diferentes soluções. O material coletado foi agitado em vórtex, acrescido a Caldo Letheem Modificado e submetido a filtração em membrana Millipore® 0,45µm. A membrana foi depositada em Tryptic Soy Ágar para crescimento microbiano. A identificação presuntiva dos micro-organismos foi realizada manualmente considerando-se aspectos morfotintoriais e reações bioquímico/fisiológicas. As variáveis foram descritas utilizando frequências, porcentagens e medidas de tendência central. O projeto foi aprovado pelo Comitê de Ética e Pesquisa da Universidade Federal de Minas Gerais (CAAE ­ 67493417.1.0000.5149). As médias das cargas microbianas na solução de detergente enzimático variaram de 19,9 UFC/mL após primeiro uso, 51,1 UFC/mL após terceiro uso e 67,1UFC/mL após o quinto reuso. Nos canais de ar/água e biópsia houve aumento de micro-organismos Gram negativos ao longo das reutilizações do detergente. Foram recuperados, Enterobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas spp., Staphyloccocus aureus, Staphyloccocus coagulase negativa. Pseudomonas spp. foi o micro-organismo mais identificado em todas as alíquotas coletadas. Verificou-se a importância da escovação do canal de biópsia para correta remoção de micro-organismos. Conclui-se que a reutilização das soluções de detergente enzimático contribuiu para contaminação dos aparelhos endoscópicos com micro-organismos potencialmente patogênicos. Faz-se necessário a reavaliação de protocolos institucionais, no sentido de que seja cumprida a orientação da Anvisa por meio da RDC nº 55 de 14 de novembro de 2012 de que os detergentes enzimáticos não devem ser reutilizados sob perda da eficiência do produto. As características físico químicas dos detergentes enzimáticos devem ser respeitadas pelos serviços de saúde conforme parâmetros estabelecidos pelos fabricantes.(AU)

In Brazil, it is recommended that during the cleaning of Health Products the detergent used has enzymatic action. Although Collegiate Board Resolution No. 55 of November 14, 2012 of the National Agency of Sanitary Surveillance advises against the reuse of this cleaning solution, it is known that in clinical practice they are reused several times for immersion of health products, such a gastrointestinal endoscope, which may compromise the effectiveness of the enzymatic detergent action and thus the safety in the processing. This research aimed to evaluate the microbial load present in the enzymatic detergent solution during its reuse in the manual cleaning of endoscopic gastrointestinal devices. This was a cross-sectional study performed at a gastrointestinal endoscopy service at a university hospital in Belo Horizonte and at the Oral Microbiology and Anaerobic Laboratory of ICB/UFMG. The sample consisted of 57 endoscopes and 76 aliquots of enzymatic detergent solution collected from several replicates of 19 different solutions. The collected material was vortexed, added to Modified Letheem Broth and subjected to Millipore® 0.45 µm membrane filtration. The membrane was deposited in Tryptic Soy Ágar for microbial growth. The identification of the microorganisms was performed manually considering morphotintorial aspects and biochemical/physiological reactions. The variables were described using frequencies, percentages and measures of central tendency. The project was approved by the Ethics and Research Committee of the Federal University of Minas Gerais (CAAE - 67493417.1.0000.5149). The mean values of the microbial loads in the enzymatic detergent solution varied from 19.9 UFC/mL after first use, 51.1 UFC/mL after third use and 67.1 UFC/mL after the fifth reuse. In the air/water and biopsy channels there was an increase of Gram negative microorganisms along the reuse of the detergent. Enterobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas spp., Staphylococcus aureus, Coagulase-negative Staphyloccocus were recovered. Pseudomonas spp. was the most identified microorganism in all aliquots collected. It was verified the importance of brushing the biopsy channel for correct removal of microorganisms. It was concluded that the reuse of enzyme detergent solutions contributed to the contamination of the endoscopes with potentially pathogenic microorganisms. It is necessary to re-evaluate institutional protocols, in order to comply with Anvisa's guidance through RDC nº. 55 of November 14, 2012 that enzymatic detergents should not be reused under loss of product efficiency. The physical characteristics of the enzymatic detergents must be observed by the health services according to the parameters established by the manufacturers.(AU)

Microbiological analysis of bile in patients with benign and malignant biliopancreatic diseases and its consequences / Análise microbiológica da bile em pacientes com doenças biliopancreáticas benignas e malignas e suas consequências

ABSTRACT Background Bactibilia has several consequences to human health. Objetive Assessing the bile microbiology of patients with biliopancreatic diseases in order to identify bacteria and their possible infectious complications. Methods Retrospective study of 30 bile culture samples from patients with benign and malignant biliopancreatic diseases. The samples were assessed to set the bile microbiological flora and to search for its possible link with comorbidity, carcinogenesis and postoperative infectious complications. Results Thirty bile samples from patients at mean age ≈57.7 years, mostly female (n=18), were assessed. Bactibilia was found in 12 cases, mostly in patients with benign diseases (n=8), older than 50 years (n=23) and female (n=10). Adenocarcinoma of the duodenal papilla (n=9) and cholelithiasis (n=8) were the most common diseases. Escherichia coli (n=5) and Klebsiella sp (n=3) were predominantly found in patients with benign diseases; and Klebsiella sp (n=2) and Streptococcus sp (n=2) were prevalent in cancer patients. There were postoperative infectious complications in seven cases, five of them in bactibilia-associated patients (P=0.084). Conclusion Bactibilia was found in 12 samples and Escherichia coli and Klebsiella sp were most often identified in patients with benign diseases, as well as Streptococcus sp and Klebsiella sp in cancer patients. There was a trend of higher postoperative infectious complication incidence in patients with bactibilia.

RESUMO Contexto Bacteriobilia pode produzir várias consequências para a saúde humana. Objetivo Avaliar a microbiologia da bile de pacientes com doenças biliopancreáticas para identificar bactérias e possíveis consequências. Métodos Estudo retrospectivo microbiológico. Trinta amostras de bile de pacientes com doenças biliopancreáticas benignas e malignas foram avaliadas para determinar a flora microbiológica da bile e procurar alguma possível relação dessa com comorbidades, carcinogênese e complicações infecciosas pós-operatórias. Resultados As amostras de bile foram avaliadas em pacientes, com idade média ≈57,7 anos, a maioria mulheres (n=18). Evidenciou-se bacteriobilia em 12 casos, a maioria em pacientes com doenças benignas (n=8); pacientes com mais de 50 anos (n=23) e mulheres (n=10). As doenças mais comuns foram o adenocarcinoma de papila duodenal (n=9) e a colelitíase (n=8). Escherichia coli (n=5) e Klebsiella sp (n=5) foram as bactérias mais identificadas em pacientes com doenças benignas; sendo a Klebsiella sp (n=2) e o Streptococcus sp (n=2) as que predominaram nos pacientes com cânceres. As complicações pós-operatórias exclusivamente infecciosas ocorreram em sete casos, sendo em cinco desses associados à bacteriobilia (P=0,084). Conclusão Bacteriobilia foi evidenciada em 12 amostras, sendo as bactérias mais identificadas Escherichia coli e Klebsiella sp em pacientes com doenças benignas; e Streptococcus sp e Klebsiella sp nos pacientes com câncer. Existiu uma tendência a maior incidência de complicações infecciosas pós-operatórias em pacientes com bacteriobilia.

Urinary tract infection and indwelling urinary catheters: prospective study in gynecological surgery with antibiotic prophylaxis / Infecção do trato urinário e cateter vesical de demora: um estudo prospectivo em cirurgia ginecológica com antibioticoprofilaxia

ABSTRACT CONTEXT AND OBJECTIVES: Urinary tract infections are the most common cause of hospital-acquired infections, and the use of indwelling urinary catheters is a predisposing factor for their development. The aims of this study were to estimate the frequency of pre and postoperative bacteriuria, identify the microorganisms involved, count the colony-forming units, determine the antibiotic sensitivity profile and compare the results from pre and postoperative urinalyses among women undergoing gynecological surgery with implantation of a urinary catheter. DESIGN AND SETTING: Non-controlled prospective observational single-cohort epidemiological study carried out at a university hospital. METHODS: Urine samples were collected before and 24 hours after catheterization for urinalysis, culturing and antibiotic sensitivity testing. Pre and postoperative urinalyses were compared using Wilcoxon and McNemar non-parametric tests. RESULTS: Fifty-one women participated in the study. Escherichia coligrew in six preoperative samples (11.8%) and Klebsiella pneumoniae in one (1.9%), but bacterial growth did not occur in any postoperative sample. Urinalysis showed lower number of pus cells in the postoperative urine samples (P < 0.05). There were no differences in red blood cell counts or in the nitrite and leukocyte esterase tests, between the samples. CONCLUSION: Bacteriuria was found in 13.7% of the preoperative samples. Gram-negative bacteria sensitive to most antibiotics were identified. In the postoperative samples, no bacterial growth was observed. Urinalysis only showed significant reduction of leukocyturia in the postoperative period.

RESUMO CONTEXTO E OBJETIVOS: As infecções urinárias são a causa mais comum de infecções hospitalares, e o uso de cateteres de demora é fator predisponente para o seu desenvolvimento. Os objetivos deste estudo foram estimar a frequência de bacteriúria pré e pós-operatória, identificar os germes encontrados, a contagem de unidades formadoras de colônias e o perfil de sensibilidade aos antibióticos, além de comparar os resultados dos exames de urina pré- e pós-operatórios em mulheres submetidas a cirurgias ginecológicas com cateterismo vesical. TIPO DE ESTUDO E LOCAL: Estudo epidemiológico, observacional, de coorte única, prospectivo, não controlado, realizado em hospital universitário. MÉTODOS: Amostras de urina foram colhidas antes da cateterização e após 24 horas para urinálise, cultura e antibiograma. Os resultados da urinálise no pré- e pós-operatório foram comparados utilizando-se os testes não paramétricos de Wilcoxon e McNemar. RESULTADOS: Participaram do estudo 51 mulheres. Houve crescimento de Escherichia coli em seis amostras pré-operatórias (11,8%) e deKlebsiella pneumoniae em uma (1,9%), mas não houve crescimento bacteriano em nenhuma amostra pós-operatória. A urinálise mostrou menor quantidade de piócitos na amostra de urina pós-cirúrgica (P < 0,05). Não houve diferença quanto ao número de hemácias e às reações para nitrito e leucocitesterase, entre as amostras. CONCLUSÃO: Houve bacteriúria em 13,7% das amostras pré-operatórias, sendo identificadas bactérias Gram-negativas sensíveis à maioria dos antibióticos. Não foi observado crescimento bacteriano nas amostras pós-operatórias. A urinálise mostrou somente redução significativa da leucocitúria no pós-operatório.

Abundant immunohistochemical expression of dopamine D2 receptor and p53 protein in meningiomas: follow-up, relation to gender, age, tumor grade, and recurrence

Meningiomas are common, usually benign tumors, with a high postoperative recurrence rate. However, the genesis and development of these tumors remain controversial. We aimed to investigate the presence and implications of a mutated p53 protein and dopamine D2 receptor in a representative series of meningiomas and to correlate these findings with age, gender, tumor grade, and recurrence. Tumor tissue samples of 157 patients diagnosed with meningioma (37 males and 120 females, mean age 53.6±14.3 years) who underwent surgical resection between 2003 and 2012 at our institution were immunohistochemically evaluated for the presence of p53 protein and dopamine D2 receptor and were followed-up to analyze tumor recurrence or regrowth. Tumors were classified as grades I (n=141, 89.8%), II (n=13, 8.3%), or grade III (n=3, 1.9%). Dopamine D2 receptor and p53 protein expression were positive in 93.6% and 49.7% of the cases, respectively. Neither of the markers showed significant expression differences among different tumor grades or recurrence or regrowth statuses. Our findings highlight the potential role of p53 protein in meningioma development and/or progression. The high positivity of dopamine D2 receptor observed in this study warrants further investigation of the therapeutic potential of dopamine agonists in the evolution of meningiomas.

Perfil etiológico y susceptibilidad antimicrobiana del primer episodio de infección urinaria febril / Etiologic and antibiotic susceptibility profile of the first episode of febrile urinary tract infection

Urinary tract infection (UTI) is a common infection in childhood; its diagnosis involves performing a urine culture. Aim: To describe the etiology and bacterial susceptibility of the first episode of UTI in children presenting with fever to the emergency room. Patients and Methods : One hundred and five children (2 months -5 years old) seen at the Hospital Dr. Sotero del Rio in Santiago, between November 2009 and November 2010 were evaluated. A urine specimen was obtained by transurethral catheterization. Urine was cultured and microorganisms were identified and tested for antimicrobial susceptibility. Results: 76.2% (80) of patients were women and 80% (84) were under 18 months. Urine sediment was abnormal in 82.5%. The most frequently isolated microorganism was Escherichia coli (96.1%) showing high susceptibility to aminoglycosides (near 100%), third generation cephalosporins, ciprofloxacin and nitrofurantoin; and low susceptibility to cephalothin (69%) and trimethoprim/sulfamethoxazole (66%). We found one ESBL-producing strain. Conclusion: The most common uropathogen was E. coli with good in vitro susceptibility to aminoglycosides and third generation cephalosporins, which are the recommended initial empirical therapy. E. coli ESBL-producing strains appear as emerging pathogens in community acquired UTIs in children.

La infección del tracto urinario (ITU) es muy frecuente en la niñez y su diagnóstico implica la realización de urocultivo. Objetivo: Describir la etiología y susceptibilidad bacteriana del primer episodio de ITU en niños que consultaron por fiebre en una unidad de emergencia. PacientesyMétodos: Se evaluaron 105 niños (2 meses -5 años) consultantes en la Unidad de Emergencia Infantil del Hospital Sótero del Río del área sur-oriente de Santiago entre noviembre de 2009 y noviembre de 2010, con muestra de orina tomada por cateterismo trans-uretral para sedimento de orina, urocultivo y antibiograma. Resultados: El 76,2% (80) de los pacientes fueron mujeres y 80% (84) tenía menos de 18 meses. El sedimento de orina resultó alterado en 82,5%. El microorganismo aislado con mayor frecuencia fue Escherichia coli (96,1%) que mostró buena susceptibilidad in vitro (cercana a 100%) para aminoglucósidos, cefalosporinas de tercera generación, quinolonas y nitrofurantoína, y baja susceptibilidad para cefalotina (69%) y cotrimoxazol (66%). Una cepa era productora de β-lactamasa de expectro extendido (bLEE). Conclusión: El uropatógeno más frecuente fue E. coli que demostró buena susceptibilidad in vitro a aminoglucósidos y cefalosporinas de tercera generación, antimicrobianos parenterales recomendados como tratamiento empírico inicial para este grupo de pacientes. Las cepas de E. coli productoras de bLEE aparecen como patógenos emergentes en las ITUs adquiridas por niños en la comunidad.

Qualidade microbiológica de manteiga de garrafa comercializada no município de Petrolina, PE / Microbiological quality of bottled butter commercialized in the city of Petrolina, PE

A manteiga de garrafa é um alimento muito apreciado nos estados do Nordeste do Brasil, onde sua produção ocorre predominantemente de forma artesanal. Nesse trabalho, objetivou-se avaliar a qualidade microbiológica da manteiga de garrafa produzida e comercializada no município de Petrolina, PE.

Phenotypic & molecular characterization of AmpC β-lactamases among Escherichia coli, Klebsiella spp. & Enterobacter spp. from five Indian Medical Centers.

Background & objectives: AmpC β-lactamases which are often plasmid mediated hydrolyze all β-lactam antibiotics except cefepime and carbapenems. We evaluated the presence of AmpC β-lactamases among Enterobacteriaceae strains recovered prospectively from patients at five Indian tertiary care centres. Methods: The study included 909 consecutive Gram-negative isolates recovered from clinically significant specimens during June 2007 - May 2008 as part of an ICMR-ESBL study. Among the study isolates, 312 were found to be cefoxitin resistant by disc diffusion test (DDT). Minimum inhibitory concentration (MIC) determination by E test was done against amikacin, levofloxacin, impinem, meropenem, ertapenem, tigecycline and piperacillin-tazobactam. Combined DDT using phenyl boronic acid as inhibitor with cefoxitin was used for phenotypic confirmation of AmpC phenotype. The common Amp C genotypes ACC, FOX, MOX, DHA, CIT and EBC were detected by multiplex PCR. Results: Plasmid mediated Amp C phenotype was confirmed in 114 of the 312 (36.5%) cefoxitin resistant isolates with 255 (81.7%) showing multidrug resistance. Susceptibility to tigecycline was highest (99%) followed by imipenem, meropenem (97%), ertapenem (89%), amikacin (85%), and piperacillin-tazobactam (74.6%). Levofloxacin resistance was 82 per cent. ESBL co carriage was observed among 92 per cent of Amp C producers. Among 114 Amp C producers, 48 could be assigned a genotype, this included CIT- FOX (n=25), EBC (n=10), FOX (n = 4), CIT (n=3), EBC-ACC (n=2) and one each of DHA, EBC-DHA, FOX -DHA and FOX-EBC-DHA. Interpretation & Conclusions: Overall, AmpC phenotypes were found in 12.5 per cent isolates, multidrug resistance and ESBL co-carriage among them was high suggesting plasmid mediated spread. The study results have implications in rational antimicrobial therapy and continued surveillance of mechanisms of resistance among nosocomial pathogens.

A case of necrotizing pancreatitis subsequent to transcatheter arterial chemoembolization in a patient with hepatocellular carcinoma

Necrotizing pancreatitis is one of the rare complications of transcatheter arterial chemoembolization (TACE). Necrotizing pancreatitis after TACE may result from the development of ischemia caused by regurgitation of embolic materials into the vessels supplying the pancreas. We report a case of post-TACE necrotizing pancreatitis with abscess formation in a patient with hepatocellular carcinoma. The patient had suffered hepatic artery injury due to repetitive TACE; during his 25th TACE procedure he had submitted to selective catheterization of the feeding vessel from the dorsal pancreatic artery with a cytotoxic agent and Gelfoam particles. The patient complained of abdominal pain after the TACE procedure, and a CT scan led to a diagnosis of necrotizing pancreatitis with abscess formation. The pancreatic abscess progressed despite general management of the pancreatitis, including antibiotics. Percutaneous catheter drainage was performed, and the symptoms of the patient improved.

Frequência e perfil de resistência de Klebsiella spp. em um hospital universitário de Natal/RN durante 10 anos / Frequence and resistance profile of Klebsiella spp. isolates in a university hospital in Natal/RN during a ten-year period

INTRODUÇÃO: As espécies de Klebsiella spp. podem causar vários tipos de infecções, principalmente hospitalares, e têm merecido destaque pelos seus variados e emergentes mecanismos de resistência. OBJETIVOS: Determinar a frequência de isolamento e a caracterização do perfil de resistência de Klebsiella spp. em um hospital universitário durante um período de 10 anos e, ainda, avaliar a tendência para o crescimento dessa resistência. Material e método: Fez-se um estudo descritivo e retrospectivo a partir de dados coletados nos livros de registro do Laboratório de Microbiologia Clínica do hospital investigado, correspondentes ao período de janeiro de 1999 a dezembro de 2008. RESULTADO: A frequência de isolamento de Klebsiella spp. foi de 13,4 por cento com predominância em uroculturas (56,4 por cento). Houve aumento significativo na resistência para a maioria dos antimicrobianos testados ao longo do período analisado com tendência para o crescimento da mesma. Nesse período, isolou-se 23 por cento de Klebsiella spp. com fenótipo produtor de betalactamases de amplo espectro (ESBL). DISCUSSÃO: O isolamento de Klebsiella spp. resistente a antimicrobianos em amostras de origem clínica e a detecção da tendência do crescimento da resistência, inclusive às drogas de reserva terapêutica, são motivos de grande preocupação. Nesse hospital, a implantação de métodos de triagem e de confirmatórios para os mecanismos de resistência de Klebsiella spp. poderiam auxiliar no diagnóstico e no tratamento das infecções causadas por esse microrganismo. CONCLUSÃO: A tendência de crescimento na resistência aos antibióticos detectada neste estudo reforça a importância de monitoramentos contínuos. Estes elucidam características locais, orientando para melhores medidas de controle.

INTRODUCTION: Klebsiella spp. species can cause several infections, particularly nosocomial ones. Furthermore, its multiple emerging resistance mechanisms have been widely described. OBJECTIVES: To determine the isolation frequency and resistance profile of Klebsiella spp. at a university hospital during a ten-year period as well as to assess the increase in its resistance. Material and method: A retrospective and descriptive study was carried out based on data collected from the record books of the Laboratory of Clinical Microbiology of the investigated Hospital from January 1999 to December 2008. RESULTS: The isolation frequency of Klebsiella spp. was 13.4 percent, predominantly in urine cultures (56.4 percent). There was a significant increase in resistance to most antimicrobials tested over the analyzed period; 23 percent of Klebsiella spp. with ESBL phenotype was isolated over this period. DISCUSSION: Multi-resistant Klebsiella spp. isolates from clinical samples as well as its growing trend in resistance mechanisms, including to reserve drugs, are cause for great concern. The implementation of screening and confirmatory methods of bacterial resistance could aid in the diagnosis and treatment of infections caused by this microorganism. CONCLUSION: The increase in resistance to antibiotics reinforces the importance of continuous monitoring, which elucidates local characteristics and allows more suitable control measures.

Antibióticos y expresión de betalactamasas de espectro extendido (BLEE) en agentes bacterémicos / Use of antibiotics and expression of extended spectrum betalactamases (ESBL) in bacteremic agents

Objective: To evaluate the association between Escherischia Coli (E. Coli) and Klebsiella spp bacteremia, both of which produce Extended Spectrum beta-lactamases (ESBL), and the use of third generation cephalos-porines (TGC) during 30 days prior to the development of the infection. Patients and Methods: Case study of all patients hospitalized between 2004 and 2007 at Hospital de Niños Roberto del Río, who presented E. coli y Klebsiella spp bacteremia. Results: 212 episodes of E. coli o Klebsiella spp bacteremias were found; 205 contained all necessary data, 47 of which were cases and 158 were controls. The use of TGC showed a statistically significant correlation with the finding of ESBL in those patients' bacteremias (OR 2,5; IC95 percent 1,26-5). In addition, an independent statistically significant association was found with hospital acquired infections (OR 8,2; IC95 percent 2,7-27,1), and the use of Central Venous Catheter (OR 38,9; IC95 percent 14,3-106,0) became close to statistical significance. Conclusions: This study shows a statistically significant association between ESBL bacteremias and use of TGC within 30 days prior to the infection, confirming other literature reports.

Objetivo: Estudiar la asociación entre bacteriemia por E. coli y Klebsiella spp productoras de beta-lactamasas de espectro extendido (BLEE) y el uso de cefalosporinas de tercera generación (CTG) durante los 30 días previos al desarrollo de la infección. Método: Estudio caso control encestado desarrollado entre los años 2004 y 2007 en el Hospital de Niños Roberto del Río, con análisis retrospectivo de los pacientes con bacteriemias por E. coli y Klebsiella spp. Resultados: Se registraron 212 episodios de bacteriemias por E. coli o Klebsiella spp, en 205 de ellos se contó con los registros necesarios, 47 fueron casos y 158 controles. El uso de CTG se asoció de modo estadísticamente significativo con la probabilidad de aislar cepas BLEE de bacteriemias en estos pacientes (OR 2,5; IC95 por ciento 1,26-5). Además, se observó asociación independiente y estadísticamente significativa con Infección Intrahospitalaria (OR 8,2; IC95 por ciento 2,7-27,1), y el uso de Catéter Venoso Central (OR 38,9; IC95 por ciento 14,3-106,0) se acercó a la significancia estadística. Conclusiones: Nuestro trabajo establece la asociación estadística entre bacteriemia por cepas BLEE y el uso de CTG dentro de los 30 días previos al desarrollo de la infección, confirmando lo sugerido por la literatura.

Nosocomial and community infections due to class A extended-spectrum β-lactamase (ESBLA)-producing Escherichia coli and Klebsiella spp. in southern Brazil

OBJECTIVES: To determine the prevalence of class A extended spectrum β-lactamases (ESBL)-producing Escherichia coli and Klebsiella spp., and to investigate clonality among ESBL-producing isolates of nosocomial and community infections. METHODS: The study involved 354 nosocomial infections samples and 992 community infections samples, obtained between 2003 and 2006 at Caxias do Sul, RS. The detection of ESBL was performed by the disk-diffusion test. Presence of blaCTX-M, blaSHV and blaTEM β-lactamase genes was evaluated by PCR, and genomic typing was determined by pulsed-field gel electrophoresis analysis. RESULTS: Higher frequency of ESBL-producing isolates were detected among nosocomial samples of E. coli (6.7 percent) and Klebsiella (43.7 percent), than those obtained from community infections (0.4 percent and 2.6 percent). blaTEM and blaCTX were the most prevalent ESBL gene families in both E. coli and Klebsiella isolates. Different pulsotypes were obtained among ESBL-producing E. coli and 11 clones for Klebsiella spp., which occurred over the years and in different hospital wards. Among ESBL-producing K. pneumoniae, 74.3 percent transferred ESBL genes by conjugation and exhibited concomitant decreased aminoglycosides susceptibility. CONCLUSION: ESBL-producing E. coli, and especially K. pneumoniae are essentially a nosocomial problem, and their dissemination to the community is relatively limited. The great genetic variability observed among ESBL-producing bacteria indicates polyclonal spread and high transference of ESBL genes between bacteria in the hospital environment. This information is of paramount importance for nosocomial infection control.

Detección de cepas productoras de b-lactamasas de espectro extendido por el sistema DIRAMIC: Comparación con los métodos doble difusión con discos y E-test / Detection of expanded-espectrum-betalactamases by DIRAMIC system: Comparison with the double-disk synergy test and E-test

The capacity of the DIRAMIC system to detect strains producing extended-spectrum-betalactamase (ESBL) was evaluated through the comparison with two phenotypic confirmatory tests: double-disk synergy test and E-test. Ninety seven clinical isolates of Escherichia coli y Klebsiella spp. previously characterized; not repeated and suspected of being ESBL producers were studied by the three methods. In comparison with E-test and double-disk synergy test, DIRAMIC system showed a sensitivity of 85.7 percent and 92.7 percent as well as specificity of 100 percent and 92.9 percent; respectively. The values found have a very high degree of concordance (kappa index > 0.80). The results obtained vouch for the utility of the DIRAMIC as a rapid method to alert about the presence of strains producing ESBL enzymes.

Se evaluó la capacidad del sistema DIRAMIC-Cuba para detectar cepas productoras de p-lactamasas de espectro extendido (BLEE), mediante la comparación con dos métodos fenotípicos confirmatorios: doble difusión con discos y E-test. Noventa y siete aislados clínicos de Escherichia coli y Klebsieüa spp previamente caracterizados, no repetidos, sospechosos de producir BLEE se estudiaron por los tres procedimientos para determinar sensibilidad, especificidad y concordancia entre los resultados. Para el sistema DIRAMIC se encontró una sensibilidad de 85,7 y 92,7 por ciento y una especificidad de 100 y 92,9 por ciento en comparación con E-test y doble difusión con discos, respectivamente. Los valores de concordancia encontrados fueron muy altos (índice kappa > 0,80). Los resultados obtenidos avalan la utilidad del sistema DIRAMIC como vía rápida, para alertar al médico acerca de la presencia de cepas productoras de BLEE, aunque es necesario profundizar y ampliar el estudio a modo de emitir resultados más precisos.

Caracterización de lactantes con infección del tracto urinario:2009-2010: Hospital Regional de Antofagasta / Characterization of infants with urinary tract infection: 2009-2010: Regional Hospital of Antofagasta

INTRODUCCION: Las infecciones del tracto urinario (ITU) son una de las enfermedades bacterianas más frecuentes en pediatría. La incidencia de ITU en el paciente pediátrico no se conoce con exactitud. En recién nacidos se estima en 1 por ciento con una mayor proporción de varones, mientras que en lactantes aumenta entre 3 a 5 por ciento con igual proporción en ambos sexos. OBJETIVO: Caracterizar el perfil clínico-epidemiológico de lactantes con diagnóstico de ITU en el servicio de Pediatría del Hospital Regional de Antofagasta. PACIENTES Y METODOS: Estudio descriptivo retrospectivo de corte transversal. Se analizaron fichas clínicas de pacientes de la unidad de lactantes del Servicio de Pediatría del Hospital Regional de Antofagasta con diagnóstico de ITU al egreso del periodo Mayo 2009 - Mayo 2010. La muestra final fue 88 pacientes. Los datos fueron analizados y tabulados en una planilla Excel2007. RESULTADOS: Un 71,5 por ciento fueron sexo femenino. La edad promedio fue de 8,9 meses con rango entre 1 mes y 23 meses. Los síntomas más frecuentes fueron fiebre (97,7 por ciento), vómitos (30,7 por ciento), diarrea (23,1 por ciento), inapetencia (21.6 por ciento) y llanto/irritabilidad (16 por ciento). Las alteraciones de laboratorio fueron PCR elevada (96 por ciento), piuria (94,3 por ciento), leucocitosis (87,5 por ciento) y nitritos positivos (48 por ciento). Sólo fueron consignados el 85,2 por ciento de los urocultivos, de los cuales el patógeno más frecuente fue Escherichia coli en el 84 por ciento. DISCUSION: La ITU en lactantes en nuestro estudio fue más frecuente en el sexo femenino, teniendo como signo preponderante la fiebre. El germen más común fue Escherichia coli, correlacionándose con la literatura. Valoramos la importancia de solicitar urocultivos para la confirmación microbiológica.

INTRODUCTION: Urinary tract infections (UTI) are one of the most frequent bacterial diseases in pediatrics. The incidence of UTI in pediatric patients is not known with accuracy. In newborns is estimated in 1 percent with a larger proportion in males, while in infants increase between 3 to 5 percent with the same proportion for both sex. OBJECTIVE: Characterize clinically and epidemiologically the profile of infants diagnosed with UTI in the Pediatric service of the Regional Hospital of Antofagasta. METHODS AND MATERIALS: A descriptive retrospective cross-sectional study. Medical records of patients diagnosed with UTI by the end of the period of May 2009 - May 2010 from the unit of infants in the Pediatric Service of the Regional Hospital of Antofagasta were analyzed. The final sample was 88 patients. The data was analyzed using an Excel 2007 spreadsheet. RESULTS: 71,5 percent were women. The average age was 8,9 month in a range between 0,5 and 23 month. Most frequent symptoms were fever (97,7 percent), vomit (30,7 percent), diarrhea (23,1 percent), lack of appetite (21,6 percent) and crying/irritability (16 percent). The alterations in laboratory was CRP high (96 percent), pyuria (94,3 percent) and leukocytosis (87,5 percent). Only were record the 85, 2 percent of urine culture of which the most frequent pathogenic agent was Escherichia coli in the 84 percent of the cases. DISCUSSION: The UTI in infants in our study was more frequent in women and its predominant symptom is fever. The most common germ was Escherichia coli which agree the literature. We value the importance of ask for urine culture to microbiological confirmation.

Trends in bacterial pathogens of lower respiratory tract infections.

OBJECTIVES: This study was conducted to determine the bacterial aetiology of lower respiratory tract infections in this environment as well as update the clinicians in the various antimicrobial alternatives available in the treatment. METHODS: Between September 2002 and February 2005, 157 bacterial pathogens from 556 patients with lower respiratory tract infections were isolated from sputum specimens, and subjected to susceptibility testing, using standard bacteriologic techniques. RESULTS: Out of the 556 cases, only 150 (27%) had an established bacterial aetiology. One pathogen was demonstrated in 143 (95.3%) patients and seven (4.7%) had mixed infections. The most prevalent single pathogen was Klebsiella pneumoniae (38%) while the most prevalent bacterial combination was Klebsiella and Pseudomonas species (2%). Isolates of Klebsiella pneumoniae were susceptible to ciprofloxacin, gentamicin and ceftriaxone. CONCLUSIONS: Bacteriological diagnosis and antibiotic resistance surveillance are indispensable in the effective management of lower respiratory tract infections.

Characterization of klebsiella species recovered from pneumoenteric buffalo calves

Bacteriological examination was carried out on 95 fecal samples and 95 nasal swabs collected from buffalo calves suffered from diarrhea and respiratory manifestation as well as 50 apparently healthy buffalo calves aged from 1 day up to 2 months old. The obtained results revealed the isolation of 6 strains of Klebsiella species from healthy buffalo calves while 14 isolates of Klebsiella sp. were recovered from buffalo calves suffered from respiratory manifestation and 18 isolates from diarrhoeic buffalo calves. All isolates were identified as K. pneumoniae subsp pneumoniae, K. pneumoniae subsp ozaenae and K. oxytoca. The pathogenicity and virulence of different Klebsiella isolates were studied in mice. The study proved that Klebsiella isolates causing mortality rates ranged from 30% up to 80% after intramuscular, subcutaneous and oral routes of infection in mice and death occur usually between day 4 and 21 post infection. This study suggested that the use of ELISA for detection of antibody titre in mice using LPS as coated antigen revealed its efficacy in detection of klebsiella infection and cross reactions and it was highly sensitive and specific enough for extensive evaluation in the field