Atividade in vitro de extratos e frações de Copernicia prunifera (Arecaceae) contra bactérias de importância clínica em saúde pública / In vitro activity of extracts and fractions of Copernicia prunifera (Arecaceae) against bacteria of clinical importance in public health / Actividad in vitro de extractos y fracciones de Copernicia prunifera (Arecaceae) frente a bacterias de importancia clínica en salud pública

Introdução: O aumento contínuo da resistência bacteriana aos antibióticos convencionais é um problema de importância global. Encontrar produtos como alternativas terapêuticas naturais é essencial. As plantas medicinais possuem uma composição química muito rica, que podem ser estruturalmente otimizadas e processadas em novos antimicrobianos. Objetivo: Avaliar o potencial antibacteriano frente a microrganismos humanos potencialmente patogênicos do extrato etanólico e frações de Copernicia prunifera. Metodologia: A triagem fitoquímica de plantas foi realizada usando métodos de precipitação e coloração e a atividade antibacteriana utilizando o método de difusão em disco e microdiluição em caldo contra cepas padronizadas de Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa e Staphylococcus aureus. Resultados: A triagem fitoquímica revela a presença de taninos, flavonoides, esteroides, triterpernóides, saponinas e alcaloides. Os extratos etanólico e frações da casca do caule e folhas tiveram atividade inibitória contra S. aureus e K. pneumonie com zona de inibição que variou de 7,0±1,73 a 9,33±0,58 mm pelo método de difusão em disco. Pelo método de microdiluição em caldo os extratos foram satisfatórios somente contra K. pneumoniae (CIM = 125 a 1000 µg/mL) S. aureus, P. aeruginosa e E. coli se mostraram resistentes aos testes (CIM > 1000 µg/mL). Conclusão: Esses resultados fornecem uma base para futuras investigações em modelos in vivo, para que os compostos de C. prunifera possam ser aplicados no desenvolvimento de novos agentes antimicrobianos contra K. pneumoniae.

Introduction: The continuous increase in bacterial resistance to conventional antibiotics is a problem of global importance. Finding products as natural therapeutic alternatives is essential. Medicinal plants have a very rich chemical composition, which can be structurally optimized and processed into novel antimicrobials. Objective: To evaluate the antibacterial potential against potentially pathogenic human microorganisms of the ethanolic extract and fractions of Copernicia prunifera. Methodology: Phytochemical screening of plants was performed using precipitation and staining methods and antibacterial activity using the disk diffusion and broth microdilution method against standardized strains of Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Results: Phytochemical screening reveals the presence of tannins, flavonoids, steroids, triterpernoids, saponins and alkaloids. The ethanolic extracts and fractions of stem bark and leaves had inhibitory activity against S. aureus and K. pneumonie with zone of inhibition ranging from 7.0±1.73 to 9.33±0.58 mm by disc diffusion method. By broth microdilution method the extracts were satisfactory only against K. pneumoniae (MIC = 125 to 1000 µg/mL) S. aureus, P. aeruginosa and E. coli were resistant to the tests (MIC > 1000 µg/mL). Conclusion: These results provide a basis for further investigation in in vivo models, so that compounds from C. prunifera can be applied in the development of new antimicrobial agents against K. pneumoniae.

Introducción: El continuo aumento de la resistencia bacteriana a los antibióticos convencionales es un problema de importancia mundial. Es esencial encontrar productos como alternativas terapéuticas naturales. Las plantas medicinales tienen una composición química muy rica, que puede optimizarse estructuralmente y transformarse en nuevos antimicrobianos. Objetivo: Evaluar el potencial antibacteriano frente a microorganismos humanos potencialmente patógenos del extracto etanólico y fracciones de Copernicia prunifera. Metodología: Se realizó el cribado fitoquímico de las plantas mediante los métodos de precipitación y tinción y la actividad antibacteriana mediante el método de difusión en disco y microdilución en caldo frente a cepas estandarizadas de Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa y Staphylococcus aureus. Resultados: El cribado fitoquímico revela la presencia de taninos, flavonoides, esteroides, triterpernoides, saponinas y alcaloides. Los extractos etanólicos y las fracciones de la corteza del tallo y las hojas presentaron actividad inhibitoria contra S. aureus y K. pneumonie con una zona de inhibición que osciló entre 7,0±1,73 y 9,33±0,58 mm por el método de difusión en disco. Por el método de microdilución en caldo, los extractos sólo fueron satisfactorios frente a K. pneumoniae (CMI = 125 a 1000 µg/mL). S. aureus, P. aeruginosa y E. coli fueron resistentes a las pruebas (CMI > 1000 µg/mL). Conclusiones: Estos resultados proporcionan una base para futuras investigaciones en modelos in vivo, de modo que los compuestos de C. prunifera puedan aplicarse en el desarrollo de nuevos agentes antimicrobianos contra K. pneumoniae.

Infection related to Klebsiella pneumoniae producing carbapenemase in renal transplant patients / Infección relacionada con Klebsiella pneumoniae productora de carbapenemasa en pacientes trasplantados renales

ABSTRACT Objective: To evaluate the risk factors related to Klebsiella pneumoniae carbapenemase infection after renal transplantation. Methods: This was a retrospective epidemiological (case-control) study, conducted from October 2011 to march 2016. Transplanted patients with infection by this bacteria during hospitalization were selected as cases. The controls were paired by age, sex, type of donor and transplant time. The proportion of cases and controls was 1:2. Results: Thirty hundred and five patients were included in the study (45 cases and 90 controls). The risk factors found for infection by KPC were: time of hospitalization after the transplant (OR: 4.82; CI95% 2.46-9.44), delayed kidney function (OR: 5.60; CI95% 1.91-11.01) and previous infectious for another microorganism ( OR: 34.13 CI95% 3.52-132.00). Conclusion: The risk of acquisition of this bacterium was directly related to invasive procedures and exposure to the hospital environment. The findings reinforce the importance of prevention measures and control of infection by this microorganism.

RESUMEN Objetivo: Evaluar los factores de riesgo relacionados con la infección por Klebsiella pneumoniae carbapenemasa después del trasplante renal. Método: Estudio retrospectivo epidemiológico (caso-control), realizado de octubre de 2011 a marzo de 2016. Pacientes transplantados con infección por esa bacteria durante la internación fueron seleccionados como casos. Los controles se parearon por edad, sexo, tipo de donante y tiempo de trasplante. La proporción de casos y controles fue de 1: 2. Resultados: Treinta y cinco pacientes fueron incluidos en el estudio (45 casos y 90 controles). Los factores de riesgo para la infección encontrados por KPC fueron: tiempo de hospitalización después del trasplante (OR: 4,82, IC95% 2,46-9,44), función renal retardada (OR: 5,60, IC95% 1, 91-11,01) y anterior infecciosa para otro microorganismo (OR: 34,13 IC95% 3,52-132,00). Conclusión: El riesgo de adquisición de esta bacteria estuvo directamente relacionado a procedimientos invasivos y exposición al ambiente hospitalario. Los hallazgos refuerzan la importancia de medidas de prevención y control de la infección por ese microorganismo.

Successful medical drainage and surgical treatment for vertebral osteomyelitis and bilateral psoas abscess with gas formation caused by klebsiella pneumoniae in a diabetic patient

SUMMARY OBJECTIVE: We describe the case of a diabetic patient who developed vertebral osteomyelitis and bilateral psoas abscess with gas formation due to klebsiella pneumoniae. METHODS: A 64-year-old woman with a 4-year history of type-2 diabetes mellitus was admitted to the Emergency Department. The subject had a 2-day history of high-grade fever associated with chills and a 5-hour history of consciousness. She received empirical treatment with febrifuge, after which her fever decreased. RESULTS: Her fever recurred after an interval of three hours. A computed tomography scan of the abdomen revealed vertebral osteomyelitis and bilateral psoas muscle abscess with gas formation. Blood culture and purulent fluid described the growth of the Klebsiella pneumoniae. The patient received antibiotic therapy and bilateral drainage therapy after the drainage catheter was placed into the abscess cavity by CT-guidance. Due to the serious damage to the vertebral column and permanent pain, the patient underwent minimally invasive internal spinal fixation and recovered successfully. CONCLUSION: A case of vertebral osteomyelitis and bilateral psoas abscess with gas formation caused by Klebsiella pneumoniae in a diabetic patient. Antibiotic therapy, drainage, and minimally invasive internal spinal fixation were performed, which enabled a good outcome.

RESUMO OBJETIVO: Descrever o caso de uma paciente diabética que desenvolveu osteomielite vertebral e abcesso bilateral do psoas com formação de gás causada por klebsiella pneumoniae. MÉTODOS: Uma mulher de 64 anos de idade, com 4 anos de histórico de diabetes mellitus tipo 2, foi admitida no Serviço de Emergência. A paciente apresentava um quadro de dias de febre alta acompanhada de calafrios e um histórico de 5 horas de consciência. Ela recebeu tratamento empírico com antitérmico, após o qual a febre diminuiu. RESULTADOS: A febre retornou após um intervalo de três horas. Uma tomografia computadorizada do abdome revelou osteomielite vertebral e abcesso bilateral do músculo psoas com formação de gás. A cultura do sangue e o fluido purulento revelaram o crescimento de Klebsiella pneumoniae. A paciente recebeu antibióticos e terapia de drenagem bilateral após o cateter de drenagem ser posicionado na cavidade do abscesso com auxílio de TC. Devido a sérios danos à coluna vertebral e a dor permanente, a paciente foi submetida à fixação vertebral interna minimamente invasiva e recuperou-se com sucesso. CONCLUSÃO: Um caso de osteomielite vertebral e abscesso do psoas bilateral com a formação de gás causada por Klebsiella pneumoniae em uma paciente diabética. Antibioticoterapia, drenagem e fixação vertebral interna minimamente invasiva foram realizadas, o que permitiu um bom resultado.

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

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

Prevalencia de factores de riesgo para infección por Klebsiella pneumoniae resistente a carbapenémicos en adultos en un hospital de cuarto nivel, Bogotá / Prevalence of risk factor for Klebsiella pneumoniae carbapenemic-resistant infection in adults in a fourth level hospital, Bogota

Introducción: Las infecciones por gram negativos adquiridas en el hospital son una causa importante de muerte, Klebsiella pneumoniae es una de las bacterias más significativas, está además relacionada con la aparición de nuevos mecanismos de resistencia antibiótica. Los carbapenémicos han permitido combatir ciertas de estas cepas resistentes, sin embargo, la aparición de algunas con resistencia a estos antibióticos representa un reto terapéutico. A partir de este contexto clínico se planteó establecer la prevalencia de factores de riesgo en pacientes adultos con aislamientos de Klebsiella pneumoniae resistente a carbapenémicos. Material y métodos: Se realizó un estudio descriptivo retrospectivo en el Hospital Universitario Clínica San Rafael de Bogotá (Colombia). Selección de pacientes: A partir de 295 historias clínicas revisadas de pacientes hospitalizados con aislamiento positivo para Klebsiella pneumoniae con perfil de resistencia a carbapenémicos confirmada en el periodo enero 2013 a diciembre 2015, se obtuvieron 84 pacientes que cumplían todos los criterios de inclusión. Mediciones: Se tuvieron en cuenta variables demográficas como sexo y edad, así como las diferentes condiciones probablemente asociadas a la condición infecciosa. Métodos analíticos: A las variables cualitativas se les calcularon frecuencias absolutas y relativas, y a las variables cuantitativas medidas de tendencia central y de dispersión. Resultados: De 84 pacientes, el 55% correspondía a sexo femenino, tenían una edad media de 66 años. El aislamiento microbiológico en orina fue del 30.4%, en sangre 24.3%, liquido peritoneal 20.7% y secreción traqueal 12.1%. El 91,6% de la población tenía hospitalización en los últimos 3 meses, 86.5% recibió tratamiento antibiótico durante la hospitalización, de estos el 59.1% fue Meropenem, el 26,2% fue Ampicilina Sulbactam, el 23,9% fue Vancomicina, el 18,3% fue Piperacilina Tazobactam. El 54,7%se sometió a algún procedimiento quirúrgico 3 meses antes del aislamiento, de los cuales el 47.8% correspondía a cirugías abdominales y el 13,1% a cirugía de tórax, luego aparecían cirugía ortopédica, en sistema nervioso central y ginecológica. El 70% de los casos tenían antecedente de implante de dispositivos intravasculares, 45,2% exposición a ventilación mecánica, 24% uso de nutrición parenteral. Conclusiones: Este estudio establece las bases para proponer nuevos estudios que planteen estrategias de prevención con el objetivo de lograr disminuir las probabilidades de adquirir esta infección multirresistente.

Introduction: Gram-negative infections acquired in the hospital are an important cause of death, Klebsiella pneumoniae is one of the most significant bacteria, it is also related to the appearance of new mechanisms of antibiotic resistance. The carbapenems have allowed to fight certain of these resistant strains, however, the appearance of some with resistance to these antibiotics represents a therapeutic challenge. From this clinical context, it was proposed this research, to establish the prevalence of risk factors in adult patients with isolates of Klebsiella pneumoniae resistant to carbapenems. Material and methods: A retrospective descriptive study was carried out in the San Rafael Clinical University Hospital of Bogotá (Colombia). Selection of patients: From 295 reviewed clinical records of patients hospitalized with positive isolation for Klebsiella pneumoniae with carbapenemic resistance profile confirmed, in the period January 2013 to December 2015, 84 patients were obtained who fulfilled all the inclusion criteria. Measurements: Demographic variables such as sex and age were taken into account, as well as the different conditions probably associated with the infectious condition. Analytical methods: Qualitative variables were calculated absolute and relative frequencies, and quantitative variables measures of central tendency and dispersion were calculated. Results: Of 84 patients, 55% corresponded to females, had a mean age of 66 years. The microbiological isolation in urine was 30.4%, in blood 24.3%, peritoneal fluid 20.7% and tracheal secretion 12.1%. 91.6% of the population had hospitalization in the last 3 months, 86.5% received antibiotic treatment during hospitalization, of these 59.1% was Meropenem, 26.2% was Ampicillin Sulbactam, 23.9% was Vancomycin, 18.3% was Piperacillin Tazobactam. 54.7% underwent some surgical procedure 3 months before the isolation, of which 47.8% corresponded to abdominal surgeries and 13.1% to thoracic surgery, then they appeared orthopedic surgery, in central nervous system and gynecological. 70% of the cases had antecedents of implantation of intravascular devices, 45.2% exposure to mechanical ventilation, 24% use of parenteral nutrition. Conclusions: This research establishes the bases to propose new researches that propose prevention strategies with the objective of reducing the chances of acquiring this multiresistant infection.

Enterobacterias portadoras de KPC en un hospital universitario / KPC-producing Enterobacteriaceae in a university hospital

La resistencia a carbapenemes en enterobacterias representa una situación de alto impacto clínico debido a las limitadas opciones terapéuticas disponibles para el tratamiento de las infecciones causadas por estos microorganismos multirresistentes que cursan con altas tasas de morbilidad y mortalidad. En nuestro país, en los últimos 5 años, se observó la diseminación de aislamientos de Klebsiella pneumoniae portadores de carbapenemasa de tipo KPC pertenecientes al ST258, clon diseminado mundialmente. En nuestro hospital la incidencia de episodios fue aumentando, especialmente la infección del sitio quirúrgico y las bacteriemias. En el último bienio se observó la diseminación del mecanismo de resistencia a otras enterobacterias y probablemente a otros secuenciotipos de K pneumoniae con mayor sensibilidad a antibióticos no ß-lactámicos. Se hace necesario instaurar las correctas medidas de prevención y control para evitar la diseminación de estos patógenos.

The presence of carbapenem -resistant Enterobacteriaceae in clinical settings represents a concerning issue due to the limited therapeutic options available for the treatment of the infections caused by these multi-drug resistant bacteria which usually have high mortality rates. The spread of Klebsiella pneumoniae isolates belonging to ST258 was observed in the last 5 years in our country. In our hospital the number of episodes has grown with the years and the most prevalent infections were the surgical site and bacteremia. In the last 2 years KPC spread to other Enterobacteriaceae and probably to other STs in K pneumoniae which showed different susceptibility patterns to non ß-lactamic antimicrobials. We believe that it is vital to install the appropriate measures to prevent and control the dissemination of these microorganisms.

Absceso hepático por Klebsiella pneumoniae, asociado con bacteriemia y meningitis. Reporte de un caso / Klebsiella pneumoniae liver abscess associated with bacteremia and meningitis. Case report

El absceso hepático, continúa siendo un importante problema de salud pública. El causado por Klebsiella pneumoniae, se ha descrito en un 29% de los casos en algunas series. Con una mortalidad hasta del 11,3%, se presenta con mayor frecuencia en pacientes con diabetes mellitus, relacionándose con complicaciones a distancia, como meningitis, absceso cerebral y afección pulmonar. Se presenta el caso de un hombre sin factores de riesgo, con síndrome de absceso hepático por Klebsiella pneumoniae con diseminación hematógena dado por la presencia de bacteriemia y meningitis. Fue manejado con cefepime y drenaje percutáneo, presentando mejoría clínica total. La infección por Klebsiella pneumoniae, representa una etiología emergente como causa de abscesos hepáticos con un comportamiento clínico diferente al resto de infecciones relacionadas con este microorganismo...

Liver abscess remains a major public health problem. Some series have reportedKlebsiella pneumoniae as the causative agent of liver abscesses in up to 29% of cases. This condition has a high mortality rate, up to 11.3%, and it occurs more often in patients with diabetes mellitus, being related to disseminated complications such as meningitis, brain abscess, and pulmonary complications. We report the case of a male patient with a liver abscess caused by Klebsiella pneumoniae and no underlying risk factors, associated with hematogenous spread with the occurrence of bacteremia and meningitis. The patient was treated with cefepime and percutaneous drainage, and he completely recovered. Klebsiella pneumoniae infection represents an emerging etiological cause of liver abscesses, with a different clinical behavior compared to other infections related to this microorganism...

Potential virulence of Klebsiella sp. isolates from enteral diets

We aimed to evaluate the potential virulence of Klebsiellaisolates from enteral diets in hospitals, to support nosocomial infection control measures, especially among critical-care patients. Phenotypic determination of virulence factors, such as capsular expression on the external membrane, production of aerobactin siderophore, synthesis of capsular polysaccharide, hemolytic and phospholipase activity, and resistance to antibiotics, which are used therapeutically, were investigated in strains ofKlebsiella pneumoniae and K. oxytoca. Modular industrialized enteral diets (30 samples) as used in two public hospitals were analyzed, and Klebsiella isolates were obtained from six (20%) of them. The hypermucoviscous phenotype was observed in one of the K. pneumoniae isolates (6.7%). Capsular serotypes K1 to K6 were present, namely K5 and K4. Under the conditions of this study, no aerobactin production, hemolytic activity or lecithinase activity was observed in the isolates. All isolates were resistant to amoxicillin and ampicillin and sensitive to cefetamet, imipenem, chloramphenicol, gentamicin and sulfamethoxazole-trimethoprim. Most K. pneumoniae isolates (6/7, 85.7%) from hospital B presented with a higher frequency of resistance to the antibiotics tested in this study, and multiple resistance to at least four antibiotics (3/8; 37.5%) compared with isolates from Hospital A. The variations observed in the antibiotic resistance profiles allowed us to classify theKlebsiella isolates as eight antibiotypes. No production of broad-spectrum β-lactamases was observed among the isolates. Our data favor the hypothesis that Klebsiella isolates from enteral diets are potential pathogens for nosocomial infections.

Relapsing klebsiella pneumoniae liver abscesses in a non-asian diabetic man / Abcessos hepáticos recidivantes provocados por klebsiella pneumoniae num homem diabético não asiático

The Klebsiella pneumoniae invasive syndrome, first described in Asia, is being reported in other parts of the world. It causes liver abcsesses, with or without extrahepatic lesions. Diabetes mellitus is the most common hosts' underlying condition. It's frequent among asian people, even outside Asia, appearing to exist genetic factors, not yet established, increasing the risk. We present a case of a 55-year-old portuguese white diabetic man, who had a previous hospital admittion due to Klebsiella penumoniae pneumonia and a two-week latter diagnosis of liver abcsess. 4 years latter he was readmitted with nausea, chills, fever and myalgias. He had elevated inflammatory markers and the CT-scan showed 2 liver abscesses. Klebsiella pneumoniae was isolated in blood and antibiotics were given with complete clinical and imaging resolution. We present this case of relapsing Klebsiella pneumonia liver abscesses in a non-asian man emphasizing the growing incidence of this condition in Europe.

O síndrome invasivo provocado por Klebsiella pneumoniae, descrito pela primeira vez na Ásia, tem vindo a ser reportado em outras partes do mundo. Provoca abcessos hepáticos, com ou sem lesões extra-hepáticas associadas. A diabetes mellitus é a condição predisponente do hospedeiro mais comum. É frequente nos indivíduos asiáticos, mesmo fora da Ásia, parecendo existir factores genéticos, ainda não estabelecidos, que aumentam o risco da infecção. Apresentamos o caso de um homem português de 55 anos, diabético que tinha uma admissão prévia no hospital por pneumonia provocada por Klebsiella pneumoniae, com diagnóstico de abcesso hepático 2 semanas depois. 4 anos mais tarde, foi readmitido com um quadro clínico de nauseas, calafrios, febre e mialgias. Apresentava parâmetros inflamatórios elevados e a tomografia computorizada (TC) abdominal revelou 2 abcessos. Foi isolada Klebsiella pneumoniae em hemoculturas e foram administrados antibióticos com completa resolução clínica e imagiológica. Apresentamos este caso de abcessos hepáticos recidivantes por Klebsiella pneumoniae num homem não asiático, enfatizando a incidência crescente desta entidade na Europa.

Improvement in medication adherence and self-management of diabetes with a clinical pharmacy program: a randomized controlled trial in patients with type 2 diabetes undergoing insulin therapy at a teaching hospital

OBJECTIVE: To evaluate the impact of a clinical pharmacy program on health outcomes in patients with type 2 diabetes undergoing insulin therapy at a teaching hospital in Brazil. METHOD: A randomized controlled trial with a 6-month follow-up period was performed in 70 adults, aged 45 years or older, with type 2 diabetes who were taking insulin and who had an HbA1c level ≥8%. Patients in the control group (CG) (n = 36) received standard care, patients in the intervention group (IG) (n = 34) received an individualized pharmacotherapeutic care plan and diabetes education. The primary outcome measure was change in HbA1c. Secondary outcomes included diabetes and medication knowledge, adherence to medication, insulin injection and home blood glucose monitoring techniques and diabetes-related quality of life. Outcomes were evaluated at baseline and 6 months using questionnaires. RESULTS: Diabetes knowledge, medication knowledge, adherence to medication and correct insulin injection and home blood glucose monitoring techniques significantly improved in the intervention group but remained unchanged in the control group. At the end of the study, mean HbA1c values in the control group remained unchanged but were significantly reduced in the intervention group. Diabetes-related quality of life significantly improved in the intervention group but worsened significantly in the control group. CONCLUSION: The program improved health outcomes and resulted in better glycemic control in patients with type 2 diabetes undergoing insulin therapy. .

contribution on Coliforms causing mastitis in cows with reference to serotypes and virulence factors of E. coli isolates

Escherichia coli [E. coli] is the predominant coliform species causing intramammary infections. Where in the present study, E. coll isolates were 1.8 strains [17.82%] followed by Enterobacter aerogenes 3 strains [2.97%] and Klebsiella pneumoniae one strain [0.99%] from 101 clinical mastitic milk samples of cows. Eighteen E. coli isolates were serotyped to nine different serogroups; O111:H4 [3], O127:H6 [3], O26 [2], O126 [2], O119:H6 [1], O114:H21 [1], O55:H7 [1], O44:H18 [1], O124[1] and [3] untyped. Virulence tests were performed on the 18 isolated E. coll, it was found that 15 isolates [83.3%] were serum resistant, 13 isolates [72.2%] had Congo Red binding activity, 6 isolates [33.3%] were invasive and one isolate [5.6%] had haemolytic activity. PCR was applied to detect the presence of Shiga like toxin producing E. coll [stxl and stx2 genes] on the nine different strains[one strain for each serogroup], where stxl and stx2 were found in 8 [88.9%] and 4 [44.4%] of the nine examined strains, respectively. While stxl and stx2 genes were found together in 3 strains [33.3%]. Conclusions: E. coli isolates usually posses one or more virulence factors that may help in establishment at the infection site and subsequently causing clinical bovine mastitis

Childhood Osteomyelitis: a five-year analysis of patients with sickle cell anaemia in Port Harcourt, Nigeria

Osteomyelitis is an important cause of morbidity and mortality among sickle cell patients. The aim of this study was to determine the prevalence and pattern of osteomyelitis among children with sickle cell disease at the University of Port Harcourt Teaching Hospital, Port Harcourt, Nigeria. This was a retrospective review of all the medical records of sickle cell patients below the age of sixteen years who were admitted into the Paediatric ward of the University of Port Harcourt Teaching Hospital, Port Harcourt from January 2003 to December 2007. Those with incomplete records were excluded. A total of 187 sickle cell patients were reviewed. Mean age of the study population was 6.95 +/- 4.23. There were more males [105] than females [82] giving a male female ratio of 1.3:1. Out of the 187 subjects with sickle cell anaemia 15 had osteomyelitis which accounted for a prevalence of 0.08%. Acute osteomyelitis accounted for 100% of cases. Klebsiela pneumonia was commonest organism isolated from blood culture 5 [33.3%]. Fever, leg swelling and bone pains were the commonest mode of presentation. The Tibia bone was commonly involved 8 [53.3%].The aetiological organisms were sensitive to ceftazidine and gentamycin in 55% of the positive blood cultures. The preponderance of Klebsiella pneumonia indicates a change in the previously accepted pattern of infection in which Salmonella species were considered to be the main causative organism

K. pneumoniae: ¿La nueva "superbacteria"?: Patogenicidad, epidemiología y mecanismos de resistencia / K. pneumoniae: the new "superbacteria"?: Pathogenicity, epidemiology and resistance mechanisms

La resistencia de los microorganismos a los antibióticos es un problema cada vez creciente en salud pública. Entre estos, Klebsiella pneumoniae es un representante importante no sólo por su frecuencia como causa de infecciones asociadas al cuidado de la salud y de la comunidad, sino por los mecanismos patogénicos que posee, como la capacidad de producir cápsula, la presencia de estructuras especializadas que le permiten adherirse a las células del hospedero (pilis), y de sideróforos que le permiten obtener el hierro necesario para su desarrollo. La resistencia de Klebsiella pneumoniae a los antimicrobianos ha evolucionado de acuerdo con la aparición y uso de estas moléculas en el tratamiento de los pacientes, siendo cada vez más amplio el espectro que abarcan, el cual va desde la resistencia a la ampicilina por la producción de la ß lactamasa SHV– 1, hasta la resistencia a los carbapenemos por diversos mecanismos, pasando por la producción de las ß lactamasas de espectro extendido, denominadas así por su capacidad de degradar las cefalosporinas de últimas generaciones y el aztreonam.El laboratorio de microbiología debe seguir las recomendaciones internacionales para detectar y confirmar la presencia de estos mecanismos de resistencia en las cepas cultivadas de las muestras remitidas para su estudio, e igual que el clínico, debe interpretar de manera óptima sus resultados, de tal forma que se pueda elegir y administrar el antibiótico más apropiado para el paciente.

The antimicrobial resistance is an increasing problem of public health. Klebsiella pneumoniae has become one of the most important pathogens because it is a frequent cause of nosocomial and community acquired infections and it has pathogenicity mechanisms like capsules, adhesive properties mediated by specialized estructures (pillis) and siderophores that are capable of taking up iron, an essential factor in bacterial growth. The increase in bacterial resistance to antibiotics has evolved with the use of these in patients treatments, being increasingly wide the spectrum that they include, happening from the resistance to ampicillin by the production of betalactamase SHV– 1 to carbapenems resistance by diverse mechanisms, from the production of extendedspectrum betalactamases (ESBL) that are associated with hydrolysis of extended– spectrum cephalosporins and aztreonam. Microbiology laboratory should follow international recommendations to detect and confirm the presence of this resistance mechanism in bacteria and the clinicians should make a suitable interpretation of the results to make the better choice of the antibiotic therapy.

Variaciones fenotípicas de susceptibilidad en cepas de Klebsiella pneumoniae de origen nosocomial y su asociación con la formación de biopelículas / Phenotipic variations of susceptibility in Klebsiella pneumoniae strains of nosocomial origin and their association with biofilms formation

Klebsiella pneumoniae constituye uno de los principales patógenos oportunistas asociado con infecciones nosocomiales. La morbi-mortalidad producida por este microorganismo está relacionado en gran parte con la resistencia antimicrobiana y la capacidad de formar biopelículas. En este estudio se analizaron las variaciones fenotípicas de susceptibilidad de 50 cepas de K. pneumoniae (26, productoras de bLEE y 24, no productoras de bLEE) aisladas de pacientes pediátricos con septicemia nosocomial, recluidos en la Unidad de Alto Riesgo Neonatal del Instituto Autónomo Hospital Universitario de Los Andes, Mérida, Venezuela y su asociación con la formación de biopelículas. Las cepas fueron identificadas según los métodos microbiológicos estandarizados, los patrones de susceptibilidad se determinaron por concentración inhibitoria mínima, y la formación de biopelícula por técnicas espectrofotométricas. La mayoría de las cepas productoras de bLEE se asociaron con frecuencia a otros marcadores de resistencia, tales como: tobramicina, kanamicina, tetraciclina, cloranfenicol (38%, cada uno), estreptomicina (36%), y netilmicina (34%). El fenotipo multirresistente predominante fue el constituido por bLEE+-AMG-TET-CLF (30%). El 18% de las cepas de K. pneumoniae fue sensible a los antibióticos probados. La formación de biopelículas se observó en el 77,8% de las cepas sensibles y en el 96,2% de las resistentes. No hubo relación estadística entre las variaciones fenotípicas de susceptibilidad y la formación de biopelículas (p= >0,05). Sin embargo, ambas características en K. pneumoniae deben ser consideradas en el momento de decidir una conducta terapéutica para el tratamiento de pacientes con infección nosocomial.

Klebsiella pneumoniae is the principal opportunist pathogen associated with nosocomial infections. The morbidity and mortality produced by this microorganism is related mostly to antimicrobial resistance and its capacity to form biofilms. In this study, the phenotypic variations of susceptibility in 50 Klebsiella pneumoniae strains (26 bLEE-producer and 24 no bLEE-producer) isolated from pediatric patients with nosocomial septicemia at the High-risk Neonatal Unit of the Instituto Autónomo Hospital Universitario de Los Andes, Mérida, Venezuela and their associations with biofilm formation were analyzed. These strains were biochemically identified by standard microbiology methods. Antimicrobial susceptibility was determined by minimal inhibitory concentration and biofilm formation was detected by spectrophotometric techniques. Most of bLEE strains were frequently associated with resistance markers such as tobramycin, kanamycin, tetracycline, chloramphenicol (38% each), streptomycin (36%) and netilmicin (34%). The predominant multiresistent phenotype was composed of bLEE+-AMG-TET-CLF (30%). 18% of K. pneumoniae strains were sensible to the antibiotics tested. Formation of biofilm was observed in 77.8% and 96.2% of sensible and resistent strains, respectively. There was no statistic relation between phenotipic variations of susceptibility and biofilm formation (p= >0.05). However, both characteristics of K. pneumoniae should be evaluated to determine the therapeutic conduct to treat patients with nosocomial infection.

[Endotoxemia and death caused by klebsiella pneumoniae [k1] and enteropathogenic escherichia coli in a red deer calf [cervus elaphus maral]]

A three month-old-male red deer calf [Cervus elaphus maral] was examined post mortem for the cause of death in Arasbaran preserved area in East Azerbaijan in September 2006. The main history of the case was the lack of colostrum intake after birth. The necropsy 6 hours after death, revealed severe general congestion especially in lungs and visceral organs [liver and jejunum]. The cut surface of lungs was moist and bronchial lumina contained a large amount of frothy pinkish edema fluid. Diffuse congestion of lung and porteinous exudates was prominent in examination. Histopathological examination revealed shock lung and hepatocytes dissociation with single cell necrosis in liver. Microscopic examination was in line with shock lung and alveolar edema. No parasites were observed within red blood cells. Bacteriological cultures gave rise to gram negative cocoobacilli and further biochemical tests performed on isolated colonies revealed the presence of pure Escherichia coli in liver and Klebsiella pnemoniae in lung. Using serological tests, E. coli serotypes O20 and O114 and Klebsiella pnemoniae serotype K1 were identified in purified bacterial cultures. This report presents endotoxemia and death in a red deer calf with lack of colostrum intake and transportation stress history

Análise do tecido hepático de camundongos alimentados com dieta enteral contendo prebióticos e contaminadas com Klebsiella sp / Analysis of liver tissue of mice fed an enteral diet containing probiotics and infected with Klebsiella sp / Análisis del tejido del hígado de ratones alimentados con una dieta enteral que contiene probióticos e infectados con Klebsiella sp

Objetivo: Avaliou-se, neste trabalho, o efeito da administração de frutoligossacarídeo (FOS) e insulina, adicionados à dieta enteral, na translocação de Klebsiella sp. para o fígado de camundongos. Métodos: Camundongos albinos suiços, com quatro a seis semanas de vida divididos em três grupos. Os animais do grupo I (controle) receberam apenas dieta AIN93-G. No tratamento II, os animais receberam dieta enteral contendo FOS, insulina, antibiótico e corticóide. No tratamento III, os animais receberam antibiótico e corticóide, dieta enteral sem FOS e insulina. No quinto dia após o início de oferecimento das dietas, os animais dos tratamentos II e III foram alimentados com dieta enteral contaminada com, aproximadamente, 10 UFC/g de Klebisella pneumoniae. Dois animais de cada grupo foram eutanasiados 24h após o fornecimento da dieta contaminada. O fígado desses animais foi coletado assepticamente e mantido em solução fixadora para estudos histológicos. Resultados: Análises histológicas demonstraram características mais intensas de inflamação no fígado dos animais que foram imunodeprimidos com corticóide, alimentados com dieta enteral contaminada com K. pneumoniae e sem prebióticos, em relação aos animais imunodeprimidos que receberam dieta enteral com prebióticos contaminada com com K. pneumoniae. Conclusão: Dietas enterais contendo ingredientes prebióticos diminuíram o grau de lesões degenerativas no tecido hepático.

Objective: We evaluated in this study, the effect of a fructoligosaccharides (FOS) and insulin, added to the enteral diet on translocation of Klebsiella sp. to the liver of mice. Methods: Swiss albino mice, with four to six weeks of life divided into three groups. Animals in group I (control) received only diet AIN 93-G. In treatment II, animals received enteral diet containing FOS, insulin, antibiotics and steroids. In treatment III, animals given antibiotics and corticosteroids, enteral nutrition and FOS without insulin. On the fifth day after start offering the diets, the animals in treatments II and III were fed with enteral feeding contaminated with approximately 10 CFU/g Klebisella pneumoniae. Two animals from each group were euthanized 24 hours after the diet contaminated. The liver of these animals were collected aseptically and kept in fixative solution for histological studies. Results: Histological analysis showed features of more intense inflammation in the liver of animals that were immunosuppressed with corticosteroids, enteral feeding on contaminated with K. pneumoniae and without probiotics, for immunosuppressed animals that received enteral feeding with prebiotics-contaminated with K. pneumoniae. Conclusion: Enteral diets containing prebiotic ingredients decreased the degree of degenerative lesions in liver tissue.

Objetivo: Se evaluó en este estudio, el efecto de una fructoligosaccharides (FOS) y la insulina, sumado a la dieta enteral en la translocación de Klebsiella sp. para el hígado de ratones. Métodos: ratones albino suizo, con cuatro a seis semanas de vida dividido en tres grupos. Animales en el grupo I (control) recibió dieta sólo AIN 93-G. En el tratamiento II, los animales recibieron la dieta enteral que contiene FOS, insulina, antibióticos y esteroides. En el tratamiento III, los animales que recibieron antibióticos y corticosteroides, la nutrición enteral y FOS sin insulina. En el quinto día comenzará después de ofrecer las dietas, los animales en los tratamientos II y III fueron alimentados con la alimentación enteral contaminados con aproximadamente 10 UFC/g Klebisella pneumoniae. Dos animales de cada grupo se sacrificaron 24 horas después de la dieta contaminada. El hígado de estos animales fueron recogidos asépticamente y se mantienen en solución fijadora para estudios histológicos. Resultados: El análisis histológico demostró características de más intensa inflamación en el hígado de animales que fueron inmunodeprimidos con corticosteroides, la nutrición enteral en contaminadas con K. pneumoniae y sin probióticos, por inmunodeprimidos animales que recibieron alimentación enteral con prebióticos contaminados con K. pneumoniae. Conclusión: enteral con dietas que contenían ingredientes prebióticos redujo el grado de lesiones degenerativas en el tejido hepático.

Prevalência de sepse por bactérias Gram negativas produtoras de beta-lactamase de espectro estendido em Unidade de Cuidados Intensivos Neonatal / Prevalence of extended-spectrum beta-lactamase producing Gram-negative bacterial sepsis in a Neonatal Intensive Care Unit

OBJETIVO: Determinar a prevalência e a mortalidade de sepse neonatal por bactérias Gram negativas produtoras de beta-lactamase de espectro estendido (ESBL) em Unidade de Cuidados Intensivos Neonatal. MÉTODOS: Trata-se de um estudo retrospectivo e descritivo de 236 recém-nascidos com suspeita de sepse entre 2000 e 2004. O diagnóstico de sepse foi confirmado pela presença de sinais clínicos associada à positividade da hemocultura. A triagem para bactérias ESBL foi realizada segundo os critérios do National Committee for Clinical Laboratory Standards. RESULTADOS: 84 (36 por cento) recém-nascidos apresentaram hemocultura positiva, sendo a Klebsiella pneumoniae o agente mais prevalente (47 por cento). A análise dos neonatos com infecção por Klebsiella pneumoniae mostrou que sete destas eram ESBL, perfazendo uma taxa de infecção de 0,4 por cento. Todos os recém-nascidos com Klebsiella pneumoniae ESBL - exceto um - foram hospitalizados por mais de 21 dias e necessitaram de ventilação mecânica; todos tinham cateter central, nutrição parenteral e antibióticos de largo espectro. A mortalidade ocorreu em 36 (43 por cento) dos 84 neonatos com sepse confirmada. Dentre os óbitos, as hemoculturas mostraram Gram negativos (67 por cento) e fungos (19 por cento). Em relação à Klebsiella pneumoniae ESBL, três recém-nascidos (43 por cento) morreram. CONCLUSÕES: A prevalência de sepse por Klebsiella pneumoniae ESBL no período do estudo foi de 0,4 por cento e a mortalidade de 43 por cento. É importante a detecção e o controle da disseminação deste tipo de microrganismo por seu impacto negativo na sobrevida de recém-nascidos prematuros e/ou doentes.

OBJECTIVE: To determine the neonatal sepsis prevalence and the mortality of extended-spectrum beta-lactamase producing Gram-negative bacteria (ESBL) in a Neonatal Intensive Care Unit. METHODS: This is a descriptive and retrospective study of 236 newborn infants with sepsis suspicion from 2000 to 2004. The diagnosis was confirmed by clinical signs and positive blood culture. Screening for ESBL was carried out following the National Committee for Clinical Laboratory Standards criteria. RESULTS: Eighty-four (36 percent) neonates showed positive blood culture. Klebsiella pneumoniae was the most prevalent agent (47 percent). Among the neonates with Klebsiella pneumoniae infection, seven presented ESBL infection, with an infection rate of 0.4 percent. All the patients with one exception had length of hospital stay greater than 21 days and needed mechanical ventilation; all the newborns used central catheters, parenteral nutrition and broad-spectrum antibiotics. Among the 84 patients with confirmed sepsis, 36 (43 percent) died and their blood cultures were positive for gram-negative bacteria (67 percent) and fungous (19 percent). In relation to ESBL Klebsiella pneumoniae, three (43 percent) neonates died. CONCLUSIONS: The prevalence of sepsis by ESBL Klebsiella pneumoniae was 0.4 percent and the mortality rate was 42.8 percent. It is important to detect and to control the spread of this infectious agent with its negative impact on the survival rate of premature and/or sick newborn infants.