Diseño, estandarización e implementación de una nueva técnica en Guatemala, para el diagnóstico rápido de hemocultivos positivos, utilizando la tecnología Maldi-tof / Design, standardization and implementation of a new technique in Guatemala, for the rapid diagnosis of positive blood cultures, using Maldi-tof technology

Las enfermedades infecciosas son un problema de salud que a pesar de los avances médicos siguen cobrando vi-das en todo el mundo; como las septicemias. La presente investigación tuvo por objetivo diseñar, estandarizar e implementar un protocolo inexistente en Guatemala, para el diagnóstico rutinario de hemocultivos positivos dentro de las instalaciones del Laboratorio Clínico del Hospital General San Juan de Dios, lugar en donde se encuentra el único espectrómetro de masas de tipo Maldi-tof (Matrix Assisted Laser Desorption Ionization-Time of flight-mass spectrometry).Se utilizaron 240 muestras de pacientes de los diferentes servicios. El diagnóstico se realizó compa-rando las identificaciones obtenidas a partir de cultivos microbiológicos puros con muestras directas de botellas con caldo BHI(Brain Heart Infusion).Los resultados de las dos metodologías fueron evaluados con el diseño estadístico "apareado o emparejado en grupo". La comparación no evidenció discordancia en las identificaciones; pero sí en los tiempos de respuesta. La reducción de tiempo fue de 42.9 h para bacterias Gram positivo, 45.0 h para bacterias Gram negativo y 126.2 h para levaduras, todos a favor de identificaciones a partir de muestras directas. Con esta investigación se pretende ofrecer una nueva alternativa que permitirá brindar un diagnóstico rápido, confiable y certero a la población guatemalteca. También permitirá reducir la morbimortalidad de los pacientes con septicemias, promover el ahorro de insumos hospitalarios, disminuir el tiempo de estancia hospitalaria, ahorrar el consumo de antibióticos innecesarios y contribuir indirectamente a combatir la resistencia antimicrobiana; un problema actual de gran importancia a nivel mundial.

Infectious diseases are a health problem that despite medical advances in terms of diagnosis continue to take lives worldwide, such is the case of sepsis. The purpose of this research was to design, standardize and implement a non-existent protocol in Guatemala, for the routine diagnosis of positive blood cultures, within the facilities of the clinical laboratory of the San Juan de Dios General Hospital; where the only Maldi-tof (Matrix Assisted Laser Desorp-tion Ionization-Time of flight-mass spectrometry) type mass spectrometer is located. For this, 240 samples of positive blood cultures were used, coming from patients of the different services. The microbiological diagnosis was made by comparing the identification data obtained from pure microbiological cultures and direct samples of BHI broth (Brain Heart Infusion) bottles. The results of the two methodologies were evaluated based on "paired or matched in groups" statistical design. The Maldi-tof technique did not show disagreement regarding identification between the two types of samples; but it did in the response time. The time reduction was 42.9 h for Gram positive bacteria, 45.0 h for Gram negative and 126.2 h for yeasts, supporting identification from direct samples. This research aims to provide a new diagnostic alternative that will allow access to fast, reliable, and accurate results for the Guatemalan population. It will also help to reduce e morbidity and mortality rates of patients with sepsis, to promote hospital supplies savings, decrease the patient length of stay, save unnecessary antibiotics and indirectly contribute to combating antimicrobial resistance; a critical problem faced by the world today.

Epidemiología y patrones de sensibilidad a los antimicrobianos de microorganismos aislados de hemocultivos en niños con leucemia aguda / Epidemiology and sensitivity patterns of antimicrobials in microorganisms isolated in blood cultures of children with acute leukemia

Las infecciones bacterianas son una de las principales causas de morbimortalidad en los niños con cáncer. Nuestro objetivo fue describir y comparar las características clínicas y los microorganismos causantes de bacteriemias con su sensibilidad antimicrobiana en niños con diagnóstico de LLA y LMA. Se realizó un estudio observacional, descriptivo entre julio-2016 y junio-2018. Se incluyeron todos los episodios de bacteriemia (EpB) en pacientes de 0 a 18 años con diagnóstico de LLA y LMA. Se documentaron datos epidemiológicos y demográficos de los pacientes y datos microbiológicos de los aislamientos de hemocultivos positivos. Se utilizó stata13. Se incluyeron 258 EpB en 167 pacientes; el 55% eran varones. La mediana de edad fue 81 meses (RIC 39-130). En 215 EpB (83%) se registró la presencia de algún tipo de catéter; neutropenia en 193 EpB (75%), neutropenia severa en 98/258 EpB (38%). Se pudo determinar el foco clínico en 152 EpB (59%). Ciento diez pacientes tenían LLA y 57 LMA. En LLA predominaron las enterobacterias, en LMA los cocos gram positivos. Se observó asociación entre LMA y estreptococos del grupo Viridans (p<0,01) y entre LLA y P.aeruginosa (p 0,01). Con respecto a la sensibilidad hubo 11% y 17% de bacilos negativos multirresistentes en LLA y LMA respectivamente. Todos los estafilococos coagulasa negativos fueron meticilino resistentes. La mayoría de los pacientes tenía algún tipo de catéter y neutropenia. Se observó un predominio de enterobacterias con bajos niveles de resistencia antibiótica. Estos resultados son importantes para conocer la epidemiología local y establecer tratamientos empíricos adecuados (AU)

Bacterial infections are one of the main causes of morbidity and mortality in children with cancer. Our aim was to describe and compare the clinical features and bacteremia-causing microorganisms together with their antimicrobial sensitivity in acute lymphocytic (ALL) and acute myelocytic leukemia (AML). A descriptive observational study was conducted between July 2016 and June 2018. All episodes of bacteremia (EpB) in patients between 0 and 18 years of age with ALL and AML were included. All epidemiological and demographic data of the patients and microbiological information of the isolates of the positive blood cultures were recorded. For statistical analysis stata13 was used. Overall 258 EpB in 167 patients were included; 55% were boys. Median age was 81 months (IQR 39-130). In 215 EpB (83%) some type of catheter was involved; neutropenia was observed in 193 EpB (75%) and severe neutropenia in 98/258 EpB (38%). A clinical focus could be determined in 152 EpB (59%). Of all patients, 110 had ALL and 57 AML. The predominant micro-organisms were enterobacteria in ALL and gram-positive cocci in AML. An association was observed between AML and the viridans group of streptococci (p<0.01) and between ALL and P. aeruginosa (p 0.01). Regarding sensitivity, there were 11% and 17% of multiresistant negative bacilli in ALL and AML, respectively. All coagulase-negative staphylococci weer methicillin resistant. The majority of patients had some type of catheter and neutropenia. Predominance of enterobacteria with low levels of resistance to antibiotics was observed. These results are important to understand the local epidemiology and establish adequate empirical therapies (AU)

Infections and antimicrobial resistance in an adult intensive care unit in a Brazilian hospital and the influence of drug resistance on the thirty-day mortality among patients with bloodstream infections

Abstract INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

Microbiologic characteristics and antibiotic resistance rates of diabetic foot infections / Perfil microbiológico e de resistência bacteriana no pé diabético infectado

ABSTRACT Purpose: the purpose of this research was to identify the sociodemographic and microbiological characteristics and antibiotic resistance rates of patients with diabetic foot infections, hospitalized in an emergency reference center. Methods: it was an observational and transversal study. The sociodemographic data were collected by direct interview with the patients. During the surgical procedures, specimens of tissue of the infected foot lesions were biopsied to be cultured, and for bacterial resistance analysis. Results: the sample consisted of 105 patients. The majority of patierns were men, over 50 years of age, married and with low educational level. There was bacterial growth in 95 of the 105 tissue cultures. In each positive culture only one germ was isolated. There was a high prevalence of germs of the Enterobacteriaceae family (51,5%). Gram-negative germs were isolated in 60% of cultures and the most individually isolated germs were the Gram-positive cocci, Staphylococcus aureus (20%) and Enterococcus faecalis (17,9%). Regarding antibiotic resistance rates, a high frequency of Staphylococcus aureus resistant to methicillin (63,0%) and to ciprofloxacin (55,5%) was found; additionally, 43,5% of the Gram-negative isolated germs were resistant to ciprofloxacin. Conclusions: the majority of patients were men, over 50 years of age, married and with low educational level. The most prevalent isolated germs from the infected foot lesions were Gram-negative bacteria, resistant to ciprofloxacin, and the individually most isolated germ was the methicillin resistant Staphylococcus aureus.

RESUMO Objetivo: identificar o perfil sociodemográfico, microbiológico e de resistência bacteriana em pacientes com pé diabético infectado. Métodos: tratou-se de estudo observacional, transversal que avaliou os perfis sóciodemográfico e microbiológico de pacientes portadores de pé diabético infectado internados em Pronto Socorro de referência. Os dados sociodemográficos foram coletados por meio de entrevista. Foram colhidos, durante os procedimentos cirúrgicos, fragmentos de tecidos das lesões podais infectadas para realização de cultura/antibiograma. Resultados: a amostra foi composta por 105 pacientes. O perfil sociodemográfico mais prevalente foi o de pacientes do sexo masculino, acima dos 50 anos, casados e com baixa escolaridade. Das 105 amostras de fragmentos de tecidos colhidos para realização de cultura e antibiograma, 95 foram positivas, com crescimento de um único germe em cada um dos exames. Houve predomínio de germes da família Enterobacteriaceae (51,5%). Germes Gram-negativos foram isolados em 60,0% das culturas e os espécimes mais isolados individualmente foram os cocos Gram-positivos, Staphylococcus aureus (20,0%) e Enterococcus faecalis (17,9%). Considerando-se os perfis de resistência bacteriana, verificou-se alta taxa de Staphylococcus aureus resistente à meticilina (63,0%) e à ciprofloxacino (55,5%); verificou-se, também, que 43,5% dos germes Gram-negativos eram resistentes à ciprofloxacino. Conclusões: o perfil sociodemográfico majoritário, foi o de homens, com mais de 50 anos e com baixa escolaridade. Concluímos que os germes mais prevalentes nas lesões podais dos pacientes diabéticos foram os Gram-negativos, resistentes ao ciprofloxacino e que o germe mais isolado individualmente foi o Staphylococcus aureus resistente à meticilina.

Comparison of cerebrospinal fluid lactate with physical, cytological, and other biochemical characteristics as prognostic factors in acute bacterial meningitis / Comparação do lactato no líquido cefalorraquidiano com as características físicas, citológicas e outros marcadores bioquímicos, como fator prognostico nas meningites bacterianas agudas

ABSTRACT Bacterial meningitis (BM) is associated with a high morbidity and mortality. Cerebrospinal fluid (CSF) lactate may be used as a prognostic marker of this condition. We hypothesized that CSF lactate levels would remain elevated in participants who died of acute BM compared with those who recovered from this disease. Objective: To evaluate the potential use of lactate and other CSF biomarkers as prognostic markers of acute BM outcome. Methods: This retrospective, longitudinal study evaluated dynamic CSF biomarkers in 223 CSF samples from 49 patients who fulfilled the inclusion criteria of acute BM, with bacteria identified by CSF culturing. The participants were grouped according to outcome: death (n = 9; 18.37%) and survival (n = 40; 81.63%). All participants received appropriate antibiotic treatment. Results: In the logistic regression model, lactate concentration in the final CSF sample, xanthochromia, and CSF glucose variation between the first and last CSF samples were predictors of a poor outcome (death). In contrast, decrease in CSF white blood cell count and CSF percentage of neutrophils, increase in the percentage of lymphocytes, and normalization of the CSF lactate concentration in the last CSF sample were predictors of a good prognosis. Conclusion: The study confirmed the initial hypothesis. The longitudinal analysis of CSF lactate is an important predictor of prognosis in acute BM.

RESUMO As meningites bacterianas (MB) estão associadas à alta morbidade e mortalidade. O lactato no líquido cefalorraquidiano (LCR) pode ser usado como biomarcador de prognóstico nas MB. A hipótese desse estudo é que os níveis de lactato no LCR se mantém elevados entre pacientes com MB aguda que evoluem para óbito, ao contrário do que ocorre em pacientes com bom prognóstico. Objetivo: Avaliar o uso potencial do lactato e outros marcadores no LCR como indicador de prognóstico na MB aguda. Métodos: Foi realizado um estudo retrospectivo longitudinal da dinâmica dos biomarcadores bioquímicos, celulares e físicos no LCR. Foram analisadas 223 amostras de 49 pacientes com MB aguda com bactérias identificadas por cultura do LCR. Os participantes foram divididos em dois grupos de acordo com o desfecho: óbito (n = 9; 18,37%) e não óbito (n = 40; 81,63%). Todos os participantes receberam antibioticoterapia adequada. Resultados: No modelo de regressão logística, as variáveis que diferiram significativamente entre os dois grupos foram concentração de lactato na amostra final de LCR, xantocromia e variação da concentração de glicose entre a primeira e a última amostra de LCR. A alteração desses fatores indicou desfechos negativos (óbito), enquanto a diminuição do número de leucócitos e da porcentagem de neutrófilos, assim como a normalização da concentração de lactato no LCR foram preditores de bom prognóstico. Conclusão: O estudo confirmou a hipótese inicial. A análise longitudinal do lactato no LCR é um importante preditor de prognóstico na MB aguda.

Costos médicos directos de las infecciones del tracto urinario por bacilos Gram negativos resistentes a betalactámicos en un hospital de alta complejidad de Medellín, Colombia / Direct medical costs of urinary tract infections by Gram-negative bacilli resistant to beta-lactams in a tertiary care hospital in Medellín, Colombia

Resumen Introducción. Las infecciones del tracto urinario son muy frecuentes en el ámbito hospitalario. Debido a la aparición de la resistencia antimicrobiana, la complejidad de los procesos de atención ha aumentado y, con ello, la demanda de recursos. Objetivo. Describir y comparar el exceso de los costos médicos directos de las infecciones del tracto urinario por Klebsiella pneumoniae, Enterobacter cloacae y Pseudomonas aeruginosa resistentes a betalactámicos. Materiales y métodos. Se llevó a cabo un estudio de cohorte en una institución de tercer nivel de Medellín, Colombia, entre octubre del 2014 y septiembre del 2015. Se incluyeron los pacientes con infección urinaria, unos por bacterias sensibles a los antibióticos betalactámicos, y otros por bacterias resistentes a las cefalosporinas de tercera y cuarta generación y a los antibióticos carbapenémicos. Los costos se analizaron desde la perspectiva del sistema de salud. La información clínico-epidemiológica se obtuvo de las historias clínicas y los costos se calcularon utilizando los manuales tarifarios estándar. El exceso de costos se estimó mediante análisis multivariados. Resultados. Se incluyeron 141 pacientes con infección urinaria: 55 (39 %) por bacterias sensibles a los betalactámicos, 54 (38,3 %) por bacterias resistentes a las cefalosporinas y 32 (22,7 %) por bacterias resistentes a los carbapenémicos. El exceso de costos totales ajustado de los 86 pacientes con infecciones del tracto urinario por bacterias resistentes a las cefalosporinas y a los carbapenémicos, fue de USD$ 193 (IC95% -347 a 734) y USD$ 633 (IC95% -50 a 1.316), respectivamente comparados con el grupo de 55 pacientes por bacterias sensibles a los betalactámicos. Las diferencias se presentaron principalmente en el uso de antibióticos de amplio espectro, como el meropenem, la colistina y la fosfomicina. Conclusión. Los resultados evidenciaron un incremento sustancial de los costos médicos directos de los pacientes con infecciones del tracto urinario por bacterias resistentes a las cefalosporinas o a los carbapenémicos. Esta situación genera especial preocupación en los países endémicos como Colombia, donde la alta frecuencia de infecciones del tracto urinario y de resistencia a los betalactámicos puede causar un mayor impacto económico en el sector de la salud.

Abstract Introduction: Urinary tract infections are very frequent in the hospital environment and given the emergence of antimicrobial resistance, they have made care processes more complex and have placed additional pressure on available healthcare resources. Objective: To describe and compare excess direct medical costs of urinary tract infections due to Klebsiella pneumoniae, Enterobacter cloacae and Pseudomonas aeruginosa resistant to beta-lactams. Materials and methods: A cohort study was conducted in a third level hospital in Medellín, Colombia, from October, 2014, to September, 2015. It included patients with urinary tract infections caused by beta-lactam-susceptible bacteria, third and fourth generation cephalosporin-resistant, as well as carbapenem-resistant. Costs were analyzed from the perspective of the health system. Clinical-epidemiological information was obtained from medical records and the costs were calculated using standard tariff manuals. Excess costs were estimated with multivariate analyses. Results: We included 141 patients: 55 (39%) were sensitive to beta-lactams, 54 (38.3%) were resistant to cephalosporins and 32 (22.7%) to carbapenems. The excess total adjusted costs of patients with urinary tract infections due to cephalosporin- and carbapenem-resistant bacteria were US$ 193 (95% confidence interval (CI): US$ -347-734) and US$ 633 (95% CI: US$ -50-1316), respectively, compared to the group of patients with beta-lactam sensitive urinary tract infections. The differences were mainly found in the use of broad-spectrum antibiotics such as meropenem, colistin, and fosfomycin. Conclusion: Our results show a substantial increase in the direct medical costs of patients with urinary tract infections caused by beta-lactam-resistant Gram-negative bacilli (cephalosporins and carbapenems). This situation is of particular concern in endemic countries such as Colombia, where the high frequencies of urinary tract infections and the resistance to beta-lactam antibiotics can generate a greater economic impact on the health sector.

Hemoparasites in ticks of wild birds of Serra dos Órgãos National Park, state of Rio de Janeiro, Brazil / Hemoparasitos em carrapatos de aves silvestres do Parque Nacional da Serra dos Órgãos, estado do Rio de Janeiro, Brasil

Abstract The aim of this study is to detect the presence of tick-borne agents of genera Rickettsia, Borrelia, Babesia, Ehrlichia and Anaplasma in ticks collected from native wild birds in the state of Rio de Janeiro. Birds were captured and observed carefully to find the ectoparasites. DNA detection of hemoparasites was performed by means of the polymerase chain reaction (PCR). The sequences obtained were analyzed and their homologies were compared to the available isolates in the GenBank platform database. A total of 33 birds were captured from 20 different species, of which 14 were parasitized by Amblyomma longirostre (n = 22). There was absence of DNA from agents of the genera Babesia, Anaplasma and Ehrlichia in the evaluated samples. The phylogenetic analysis indicated that one sample had 100% identity with Rickettsia bellii (KJ534309), the other two samples showed 100% identity with Rickettsia sp. Aranha strain and strain AL (EU274654 and AY360216). The positive sample for R. bellii was also demonstrated to be positive for Borrelia sp., which presented a similarity of 91% with Borrelia turcica (KF422815). This is the first description of Borrelia sp. in ticks of the genus Amblyomma in South America.

Resumo Este trabalho teve como objetivo detectar evidências moleculares da presença de agentes dos gêneros Rickettsia, Borrelia, Babesia, Anaplasma e Ehrlichia transmitidos por carrapatos coletados de aves silvestres no estado do Rio de Janeiro. Aves foram capturadas e observadas cuidadosamente a procura de ectoparasitos. A detecção de DNA de hemoparasitos foi realizada por meio da reação em cadeia da polimerase (PCR). As sequências obtidas foram analisadas e sua homologia comparada aos isolados disponíveis na base de dados da plataforma GenBank. Foram capturadas 33 aves, de 20 espécies diferentes das quais 14 estavam parasitadas por Amblyomma longirostre (n = 22). Houve ausência de DNA de agentes dos gêneros Babesia, Anaplasma e Ehrlichia nas amostras avaliadas. A análise filogenética indicou que uma amostra apresentou 100% de identidade com Rickettsia bellii (KJ534309), as outras duas amostras apresentaram 100% de identidade com Rickettsia sp. cepa Aranha e Cepa AL (EU274654 e AY360216.). A amostra positiva para R. bellii também apresentou positividade para Borrelia sp. que apresentou similaridade de 91% com Borrelia turcica (KF422815). Esta é a primeira descrição de Borrelia sp. em carrapatos do gênero Amblyomma na América do Sul.

Microbiological analysis of contact lens cases: impact of the hospital environment / Análise microbiológica de estojos de lentes de contato: impacto do ambiente nosocomial

ABSTRACT Purpose: The aim of this study was to analyze the bacterial and fungal microbiota found in contact lens cases among two groups of patients to correlate the data on the habits of contact lens users and to evaluate whether there is a difference in the culture results between users of ridged and nonridged contact lens cases. Methods: Two groups (35 patients per group) were included, consisting of hospital workers and those who had not visited a hospital in the past 30 days, and a questionnaire regarding epidemiological data and habits related to contact lens and lens case use was administered. In addition, 140 samples collected from the right and left compartments of each lens case by swabbing the bottom of the wells were tested using bacterioscopy as well as fungal and bacterial cultures via computerized identification of the species. Results: No fungal growth was identified in any of the 70 contact lens cases; however, bacteria were found in 39 cases, and there was no statistical difference between the groups. Most bacteria (>85%) were gram-negative bacilli. Contamination inone compartment of the contact lens case elevated the risk of contamination of the other side (>80%). Moreover, contamination was statistically higher in the ridged cases than in nonridged cases (p=0.0149). Conclusion: The types of bacteria contaminating the cases are generally not seen in eye diseases associated with contact lens use, suggesting that other decisive variables are involved in eye infection from a contaminated lens or case. Fungal contamination of contact lens cases appears to be an exception. Ridged cases are commonly used by contact lens wearers and present a potential risk to eye health. In addition, the results of bacterial tests between hospital workers and those who did not visit a hospital were not significantly different.

RESUMO Objetivo: O objetivo deste trabalho foi analisar a microbiota bacteriana e fúngica encontrada em estojos de lentes de contato em dois grupos, correlacionar os dados sobre os hábitos de uso de lentes de contato e avaliar se há diferença na positividade das culturas entre os usuários estojos de lentes de contato com ranhuras e sem ranhuras. Métodos: Dois grupos foram formados, trabalhadores do hospital e pessoas que não visitaram o hospital (35 indivíduos por grupo), e um questionário foi aplicado sobre dados epidemiológicos e hábitos relacionados ao uso de lentes de contato e estojos de lentes. Além disso, 140 amostras, coletadas do compartimento direito e esquerdo de cada estojo de lente, esfregando o fundo dos mesmos, foram testadas por bacterioscopia e por culturas de fungos e bactérias, com identificação computadorizada da espécie. Resultados: Não houve crescimento fúngico em nenhum dos 70 estojos de lentes de contato, porém bactérias foram encontradas em 39; não houve diferença estatística entre os grupos. A maioria das bactérias (>85%) eram bacilos gram-negativos. Quando um compartimento estava contaminado, o risco de contaminação do outro compartimento era elevado (>80%). A contaminação foi estatisticamente maior nos estojos com ranhuras (p=0,0149). Conclusão: A contaminação dos estojos parece ocorrer por bactérias que, em geral, não são encontradas em doenças oculares associadas ao uso de lentes de contato, sugerindo que existem outras variáveis decisivas nas infecções oculares de uma lente ou estojo contaminado. Contaminação de estojos de lentes de contato com fungos parece ser uma exceção. O uso de estojos com ranhuras é uma prática comum e apresenta um risco potencial à saúde ocular. Não foram encontradas diferenças significativas nos resultados dos testes bacterianos entre trabalhadores hospitalares e pessoas que não visitaram o hospital.

Epidemiology of diabetic foot infections in a reference tertiary hospital in India

Abstract Introduction The present study attempts to examine the microbial profile and antibiotic susceptibility of diabetic foot infections in the intensive care unit of a tertiary referral centre for diabetic foot. As part of the study, we also attempted to find the prevalence of blaNDM-like gene among carbapenem-resistant gram negative infections. Methodology A prospective study of 261 patients with diabetic foot infections was performed during the period between January 2014 and June 2014. Results A total of 289 isolates were obtained from 178 tissue samples from 261 patients, 156 (59.7%) males and 105 (40.2%) females, with a mean age of 58 years (-15 years), having diabetic foot infection. No growth was seen in thirty eight (17.6%) tissue samples. Out of the total samples, 44.3% were monomicrobial and 55.7% were polymicrobial. Gram negative pathogens were predominant (58.5%). Seven of the total isolates were fungal; 0.7% showed pure fungal growth and 1.7% were mixed, grown along with some bacteria. The most frequently isolated bacteria were Staphylococcus aureus (26.9%), followed by Pseudomonas aeruginosa (20.9%). Of the 58.5% gram negative pathogens, 16.5% were Enterobacteriaceae resistant to carbapenems. Among these isolates, 4 (25%) were positive for blaNDM-like gene. Among the rest, 18.6% were carbapenem-resistant Pseudomonas, among which 4 (36.3%) were blaNDM. Among the Staphylococci, 23.7% were methicillin-resistant Staphylococcus aureus. Conclusions Our results support the recent view that gram negative organisms, depending on the geographical location, may be predominant in DFIs. There is an increase in multidrug-resistant pathogens, especially carbapenem resistance and this is creeping rapidly. We need to be more judicious while using empiric antibiotics.

A practical molecular identification of nonfermenting Gram-negative bacteria from cystic fibrosis

Abstract Identification of nonfermenting Gram-negative bacteria (NFGNB) of cystic fibrosis patients is hard and misidentification could affect clinical outcome. This study aimed to propose a scheme using polymerase chain reaction to identify NFGNB. This scheme leads to reliable identification within 3 days in an economically viable manner when compared to other methods.

Microorganismos aislados de hemocultivos en niños con cáncer y neutropenia febril de alto riesgo en cinco hospitales de Santiago, Chile, período 2012-2015 / Microorganisms isolated from blood cultures in children with cancer and high-risk febrile neutropenia from five hospitals in Santiago, Chile, 2012-2015

Resumen Antecedentes: Los microorganismos aislados de hemocultivos (HC) en pacientes con neutropenia febril (NF) varían en el tiempo, siendo necesaria su vigilancia para orientar una terapia empírica adecuada. Objetivo: Identificar microorganismos aislados de HC y su perfil de resistencia (R) a antimicrobianos en niños con cáncer y NF de alto riesgo. Método: Estudio prospectivo, multicéntrico de episodios de NF de alto riesgo en pacientes bajo 18 años de edad, de cinco hospitales en Santiago de Chile, 2012-2015. Análisis de HC positivos. Resultados: Se analizaron 206 microorganismos en 185 episodios de NF de alto riesgo con HC positivos. Los aislados principales fueron bacilos gramnegativos (BGN) (46,6%) y cocáceas grampositivas (CGP) (45,1%) y los microorganismos más frecuentes Escherichia coli (22,8%), Staphylococcus coagulasa negativa (18,0%) y Klebsiella spp (16,5%). En resistencia (R) a antimicrobianos destaca: E. coli y Klebsiella spp 4,2 y 67,6% R a cefalosporinas de tercera generación (cefotaxima/ceftriaxona) respectivamente, 10,6 y 40,6% R a ciprofloxacina y 2,1 y 26,5% a amikacina, respectivamente. S. coagulasa negativa y S. aureus 86,4% y 22,2% R a oxacilina, Streptococcus grupo viridans 71% R a penicilina. Discusión: Este estudio actualiza la etiología y el perfil de R de microorganismos aislados en HC de niños con cáncer y NF de alto riesgo, herramienta esencial para el adecuado manejo de estos pacientes.

Background: Microorganisms isolated from blood cultures (BC) in patients with febrile neutropenia (NF) vary over time, requiring systematic monitoring to guide appropriate empirical therapy. Aim: To identify microorganisms isolated from BC and their antimicrobial resistance profile in children with cancer and high risk NF. Method: Prospective, multicenter study. The analysis included episodes of high-risk FN with positive BC in children under 18 years of age treated in five hospitals in Santiago, Chile, 2012-2015. Results: A total of 206 microorganisms were analyzed in 185 episodes of high-risk FN. The main isolates were Gram negative bacilli (46.6%) and Gram positive cocci (45.1%) and the most frequent microorganisms were Escherichia coli (22.8%), coagulase negative Staphylococcus (18.0%) and Klebsiella spp. (16.5%). Escherichia coli and Klebsiella spp showed 4.2% and 67.6% resistance to third generation cephalosporins (cefotaxime/ceftriaxone), 10.6% and 40.6% resistance to fluoroquinolones (ciprofloxacin) and 2.1% and 26.5% to amikacin, respectively. Coagulase negative Staphylococcus and Staphylococcus aureus had 86.4% and 22.2% resistance to oxacillin, Streptococcus viridans group had 71% resistance to penicillin. Discussion: This study updates the etiology and resistance profile of microorganisms isolated in BC from children with cancer and high risk FN, an essential tool for the adequate management of these patients.

Prevalencia y perfil de susceptibilidad antimicrobiana en bacterias aisladas de úlceras crónicas infectadas en adultos / Prevalence and susceptibility pattern of bacteria isolated from infected chronic wounds in adult patients

Resumen Introducción: Las úlceras crónicas son un problema de salud pública, agravándose por infecciones bacterianas causadas principalmente por agentes resistentes. Objetivo: Estudiar prevalencia y perfil de susceptibilidad en bacterias aisladas de úlceras crónicas en pacientes adultos. Pacientes y Métodos: Pacientes atendidos en la Fundación Instituto Nacional de Heridas entre mayo y julio de 2014, con úlceras crónicas en extremidades inferiores con signos inflamatorios clínicos. Las muestras fueron cultivadas en aerobiosis y anaerobiosis y para la identificación bacteriana se empleó el sistema de galerías API (Biomerieux). La susceptibilidad in vitro se evaluó según el método de Kirby Bauer. Resultados: Se reclutaron 73 pacientes, entre quienes 46 presentaron úlceras infectadas, diagnosticándose 33 úlceras venosas con predominio de infección polimicrobiana y 10 úlceras de pie diabético con predominio de infección monomicrobiana (p ≤ 0,05). Se aislaron 68 cepas de los 46 pacientes con úlcera infectada. Las enterobacterias predominaron en infección monomicrobiana (p ≤ 0,05) y los demás grupos bacterianos fueron levemente más frecuentes en infección polimicrobiana. La especie prevalente fue Staphylococcus aureus (24%) seguida de Pseudomonas aeruginosa (18%). Cincuenta cepas (77%) presentaron resistencia a uno o más antibacterianos. Destacamos resistencia de S. aureus a ciprofloxacina (50%) y cefoxitina (37,5%) identificándose así resistencia a meticilina en la comunidad (SARM-AC), siendo todas sensibles a cotrimoxazol. Las enterobacterias presentaron resistencia a sensibilidad a amikacina (95,5%), P. aeruginosa evidenció resistencia a ciprofloxacina (33,3%) con alta sensibilidad a gentamicina (91,7%) y amikacina (83,3%), mientras Acinetobacter spp presentó resistencia a ciprofloxacina y ceftazidima en 60%, con 100% de sensibilidad a imipenem. Streptococcus β hemolítico presentó 50% de resistencia a clindamicina y penicilina. Conclusión: Estos datos entregan información epidemiológica de infecciones de úlceras crónicas, representando un apoyo al diagnóstico, tratamiento y manejo de esta patología.

Background: Chronic wounds are considered a public health problem that may be complicated by bacterial infections, mainly caused by resistant strains. Aim: To study the bacteria prevalence and antimicrobial susceptibility in samples from adult patients with chronic wounds. Methods: Patients treated at National Institute of Wounds Foundation between May and July 2014, with chronic ulcers in lower extremities with clinical inflammatory signs were recluted. Samples were cultured in aerobic and anaerobic atmosphere and species identification was performed by API (Biomerieux) galleries. The in vitro susceptibility was evaluated according to the Kirby Bauer method. Results: From 73 patients, 46 had infected wounds most of them were venous ulcers (33) with prevalence in polymicrobial infections and 10 with foot-diabetes ulcers with prevalence in monomicrobial infections (p ≤ 0.05). Sixty-eight strains were isolated and Enterobacteriaceae were predominant in monomicrobial infection (p ≤ 0.05) and the other groups were slightly higher in polymicrobial infection. The main species were Staphylococcus aureus (24%) followed by P. aeruginosa (18%). Fifty strains (77%) were resistant or multi-resistance. We emphasize resistance of S. aureus to ciprofloxacin (50%) and cefoxitin (37.5%), thus identifying resistance to methicillin in the community (CA-SAMR), all of which are sensitive to cotrimoxazole. Enterobacteria showed sensitivity to amikacin (95.5%), P. aeruginosa showed resistance to ciprofloxacin (33.3%) with high sensitivity to gentamicin (91.7%) and amikacin (83.3%), while Acinetobacter spp showed resistance to ciprofloxacin and ceftazidime in 60%, with 100% sensitivity to imipenem. 50% Streptococcus β hemolytic showed resistance to clindamycin and penicillin. Conclusion: These data provide epidemiological information on chronic wound infections, representing support for diagnosis, treatment and management of this pathology.

Impacto das mudanças da microbiologia na peritonite bacteriana espontânea em três diferentes períodos ao longo de 17 anos / Impact of microbiological changes on spontaneous bacterial peritonitis in three different periods over 17 years

ABSTRACT BACKGROUND: Spontaneous bacterial peritonitis is a serious complication in cirrhotic patients, and changes in the microbiological characteristics reported in the last years are impacting the choice of antibiotic used for treatment. OBJECTIVE: The aim of the present study is to evaluate the changes in the epidemiology and bacterial resistance of the germs causing spontaneous bacterial peritonitis over three different periods over 17 years. METHODS: All cirrhotic patients with spontaneous bacterial peritonitis and positive culture of ascites fluid were retrospectively studied in a reference Hospital in Southern Brazil. Three periods were ramdomly evaluated: 1997-1998, 2002-2003 and 2014-2015. The most frequent infecting organisms and the sensitivity in vitro to antibiotics were registered. RESULTS: In the first period (1997-1998) there were 33 cases, the most common were: E. coli in 13 (36.11%), Staphylococcus coagulase-negative in 6 (16.66%), K. pneumoniae in 5 (13.88%), S. aureus in 4 (11.11%) and S. faecalis in 3 (8.33%). In the second period (2002-2003), there were 43 cases, the most frequent were: Staphylococus coagulase-negative in 16 (35.55%), S. aureus in 8 (17.77%), E. coli in 7 (15.55%) and K. pneumoniae in 3 (6.66%). In the third period (2014-2015) there were 58 cases (seven with two bacteria), the most frequent were: E. coli in 15 (23.1%), S. viridans in 12 (18.5%), K. pneumoniae in 10 (15.4%) and E. faecium 5 (7.7%). No one was using antibiotic prophylaxis. Considering all staphylococci, the prevalence increased to rates of the order of 50% in the second period, with a reduction in the third period evaluated. Likewise, the prevalence of resistant E. coli increased, reaching 14%. CONCLUSION: There was a modification of the bacterial population causing spontaneous bacterial peritonitis, with high frequency of gram-positive organisms, as well as an increase in the resistance to the traditionally recommended antibiotics. This study suggests a probable imminent inclusion of a drug against gram-positive organisms in the empiric treatment of spontaneous bacterial peritonitis.

RESUMO CONTEXTO: A peritonite bacteriana espontânea é uma complicação séria em pacientes cirróticos e as alterações nas características microbiológicas relatadas nos últimos anos podem afetar a escolha do antibiótico utilizado no tratamento. OBJETIVO: Os objetivos do presente estudo são avaliar as mudanças na epidemiologia e perfil de resistência bacteriana dos germes causadores de peritonite bacteriana espontânea em três períodos diferentes ao longo de 17 anos. MÉTODOS: Todos os pacientes cirróticos com peritonite bacteriana espontânea e cultura positiva de fluido ascítico foram estudados retrospectivamente em um hospital de referência no Sul do Brasil. Foram avaliados três diferentes períodos selecionados de forma randômica: 1997-1998, 2002-2003 e 2014-2015. Os organismos infecciosos mais frequentes e a sensibilidade in vitro a antibióticos foram registados. RESULTADOS: No primeiro período (1997-1998) houve 33 casos; os mais comuns foram: E. coli em 13 (36,1%), Staphylococcus coagulase-negativo em 6 (16,7%), K. pneumoniae em 5 (13,9%), S. aureus em 4 (11,1%) e S. faecalis em 3 (8,3%). No segundo período (2002-2003), houve 43 casos, os mais frequentes foram: Staphylococus coagulase-negativo em 16 (35,5%), S. aureus em 8 (17,8%), E. coli em 7 (15,5%) e K. pneumoniae em 3 (6,7%). No terceiro período (2014-2015), houve 58 casos (sete com duas bactérias), os mais frequentes foram: E. coli em 15 (23,1%), S. viridans em 12 (18,5%), K. pneumoniae em 10 (15,4%) e E. faecium 5 (7,7%). Nenhum paciente estava usando profilaxia antibiótica. Quando considerados todos os estafilococos, a prevalência aumentou para taxas da ordem de 50% no segundo período, apresentando redução no terceiro período avaliado. Do mesmo modo, a prevalência de E coli resistente aumentou, chegando a 14%. CONCLUSÃO: Houve modificação da população bacteriana causadora de peritonite bacteriana espontânea, com alta frequência de organismos gram-positivos, bem como aumento da resistência aos antibióticos tradicionalmente recomendados. Este estudo sugere uma provável inclusão iminente de um medicamento contra organismos gram-positivos no tratamento empírico da peritonite bacteriana espontânea.

Microbiological assessment of honey in México / Calidad microbiológica de la miel en México

Honey is a product used as a natural sweetener and in several regions of Mexico and other countries it is also used as a therapeutic agent. Microbiological contamination of honey can occur during its extraction and handling. Due to the use and consumption of honey we highlighted here the importance of the assessment of its microbiological quality. One thousand nine hundred twenty samples obtained from 8 honey-producing states from Mexico were analyzed. From these samples, 40.5% (777/1920) did not comply with the NMX-036-NORMEX-2006 specification. Forty five percent (777/1920) of the samples did not comply with the mesophilic aerobic microorganism specification, neither did 17% (327/1920) of the samples with the specification for molds and 18.1% (348/1920) with the specification for yeasts. With regard to coliform bacteria, the samples contained less than 3 NMP/g. Two percent of the samples contained lactic acid bacteria (LAB), Clostridium perfringens was observed in amounts of more than 100 CFU/g. None of the samples from the different states contained more than 100 CFU/g of Staphylococcus aureus; Salmonella spp. was absent in all samples. It is important to avoid contamination sources and implement good hygienic practices in order to maintain and improve the quality of Mexican honeys since a large percentage of them are intended for export. If these honeys are intended for therapeutic use, it is necessary to ensure that they comply with all quality parameters and to apply specific treatments that guarantee the removal of any pathogen that may represent a risk to the patients's health.

La miel es un producto que se usa como edulcorante natural y en algunas regiones de México y otros países, como agente terapéutico. La contaminación de la miel con microorganismos se puede presentar durante la extracción y el manejo. Dado el uso y consumo de la miel, se consideró importante evaluar su calidad microbiológica. Se trabajó con 1.920 muestras de miel obtenidas de 8 estados productores en México. De las muestras de miel analizadas, el 40,5% (777/1.920) estuvo fuera de las especificaciones de la NMX-036-NORMEX-2006. Todas esas muestras no cumplían con las especificaciones referidas a mesofílicos aerobios, en tanto que el 17% (327/1920) no cumplían las relativas a mohos y el 18,1% (348/1920), las fijadas para levaduras. En lo que se refiere a los organismos coliformes, las muestras contenían < 3 NMP/g. Se encontró que el 2% contenía BAL; Clostridium perfringens se puso en evidencia en el 12%, con más de 100 UFC/g. En el caso de Staphylococcus aureus, ninguna muestra estuvo por arriba de 100 UFC/g. Salmonella spp. no se detectó en ninguna de las muestras analizadas. Es importante evitar las fuentes de contaminación y observar buenas prácticas de higiene para mantener y mejorar la calidad de las mieles mexicanas, ya que un buen porcentaje de ellas están destinadas a la exportación. Si estas van a ser destinadas a un uso terapéutico es necesario asegurarse de cumplir con los parámetros de calidad y dar tratamientos específicos que aseguren la eliminación de algún patógeno que pueda poner en riesgo la salud del paciente que utilice la miel.

Efficacy of removal of cariogenic bacteria and carious dentin by ablation using different modes of Er: YAG lasers

The primary objective of this in vitro study was to evaluate the efficiency of removal of cariogenic bacteria and carious dentin by ablation using two lasers: fluorescence-feedback controlled (FFC) Er:YAG laser and different pulses of Er:YAG laser based on variable square pulse technology (VSPt). The secondary objective was to measure the temperature during laser ablation of carious tissue. Seventy-two extracted human molars were used in this study. Sixty teeth with carious dentin were randomly divided into four experimental groups according to the treatment for caries removal: group 1: 400 µs (FFC group); group 2: super short pulse (SSP group, 50 µs pulse); group 3: medium short pulse (MSP group, 100 µs pulse); group 4: short pulse (SP group, 300 µs pulse) and one positive control group with no treatment. Twelve teeth without carious lesion were used as a negative control group. After caries removal, swabs were taken with cotton pellets and real-time PCR analysis was performed. During caries ablation, a thermal infrared camera was used to measure the temperature changes. In all experimental groups, specimens were free of bacterial contamination after the treatment. In the SSP, MSP and SP groups, temperatures measured during caries ablation were significantly higher compared to temperatures in the FFC group (P<0.001). In this in vitro study, laser treatment for removal of carious dentin and cariogenic bacteria was an efficient treatment modality without causing excessive temperatures that might adversely affect pulp vitality.

Genomic analysis of bifunctional Class C-Class D ß-lactamases in environmental bacteria

β-lactamases, which are found in several bacterial species and environments, are the main cause of resistance to β-lactams in Gram-negative bacteria. In 2009, a protein (LRA-13) with two β-lactamase domains (one class C domain and one class D domain) was experimentally characterised, and an extended action spectrum against β-lactams consistent with two functional domains was found. Here, we present the results of searches in the non-redundant NCBI protein database that revealed the existence of a group of homologous bifunctional β-lactamases in the genomes of environmental bacteria. These findings suggest that bifunctional β-lactamases are widespread in nature; these findings also raise concern that bifunctional β-lactamases may be transferred to bacteria of clinical importance through lateral gene transfer mechanisms.

Successful and failed mini-implants: microbiological evaluation and quantification of bacterial endotoxin

Abstract Objectives Using two groups of mini-implants (successful and failed) the objectives of this in vivo study were: to evaluate the microbial contamination by the checkerboard DNA-DNA hybridization technique and to quantify the bacterial endotoxin by the limulus amebocyte lysate assay. Material and Methods The 15 successful and 10 failed mini-implants (1.6 mm diameter × 7.0 or 9.0 mm long), placed in the maxilla and/or mandible, were obtained from 15 patients undergoing orthodontic treatment. Data were analyzed statistically by the Wilcoxon rank-sum test using the SAS software (a=0.05). Results All 40 microbial species were detected in both groups of mini-implants, with different frequencies. No differences were observed between the groups with respect to microbial complexes (blue, purple, yellow, green, orange, red and other species) and endotoxin quantification (p>0.05). Conclusion Neither microbial contamination nor endotoxin quantification was determinant for the early loss of stability of the mini-implants.

The evaluation of nosocomial infections in pediatric patients with extracorporeal membrane oxygenation support

Abstract Introduction: Extracorporeal membrane oxygenation (ECMO) has become a standard technique over the past few decades in intensive care unit (ICU). Objective: A review of pediatric patients who received ECMO support in the pediatric cardiac ICU was conducted to determine the incidence, risk factors and causal organisms related to acquired infections and assess the survival rates of ECMO patients with nosocomial infections. Methods: Sixty-six patients who received ECMO support in the pediatric cardiac ICU between January 2011 and June 2014 were included in the study. Demographic, echocardiographic, hemodynamic features and surgical procedures were reviewed. Results: Sixty-six patients received a total of 292.5 days of venoarterial ECMO support. Sixty were postoperative patients. Forty-five patients were weaned from ECMO support with an ECMO survival rate of 68.2%. The rate of infection was 116.2/1000 ECMO days. Prolonged ICU stay, duration of ventilation and ECMO were found associated with development of nosocomial infection and only the duration of ECMO was an independent risk factor for nosocomial infections in ECMO patients. Conclusion: The correction of the underlying process leading to ECMO support and shortening the length of ECMO duration together with stricter application of ECMO indications would improve the infection incidence and hospital surveillance of the patient group.

Microbiota periodontal y microorganismos aislados de válvulas cardiacas en pacientes sometidos a cirugía de reemplazo de válvulas en una clínica de Cali, Colombia / Periodontal microbiota and microorganisms isolated from heart valves in patients undergoing valve replacement surgery in a clinic in Cali, Colombia

Resumen Introducción. La periodontitis es una enfermedad infecciosa que afecta los tejidos de soporte del diente y se asocia con diferentes enfermedades sistémicas, incluida la enfermedad cardiovascular. Los estudios microbiológicos permiten detectar microorganismos a partir de muestras subgingivales y cardiovasculares. Objetivo. Describir la microbiota periodontal cultivable y la presencia de microorganismos en válvulas cardiacas de pacientes sometidos a cirugía de reemplazo valvular en una clínica de Cali. Materiales y métodos. Se analizaron 30 muestras subgingivales y de tejidos valvulares mediante cultivo en medio bifásico, agar de sangre con suplemento y agar tripticasa de soya con antibiótico. Las muestras de las válvulas se analizaron mediante reacción en cadena de la polimerasa (PCR) convencional. Resultados. Los patógenos periodontales aislados de bolsas periodontales fueron Fusobacterium ( 50 % ), Prevotella intermedia/nigrescens (40 %), Campilobacter rectus (40 %), Eikenella corrodens (36,7 %), bacilos entéricos Gram negativos (36,7 %), Porphyromonas gingivalis (33,3 %) y Eubacterium (33,3 %). Los agentes patógenos aislados de la válvula aórtica fueron Propionibacterium acnes (12 %), bacilos entéricos Gram negativos (8 %), Bacteroides merdae (4 %) y Clostridium bifermentans (4 %), y de la válvula mitral, P. acnes y Clostridium beijerinckii. La PCR convencional no arrojó resultados positivos para agentes patógenos orales y solo se detectó ADN bacteriano en dos muestras. Conclusiones. La microbiota periodontal de pacientes sometidos a cirugía de reemplazo valvular estaba conformada por especies Gram negativas que han sido relacionadas con infecciones en tejidos extraorales; sin embargo, no se encontraron agentes patógenos periodontales en los tejidos de las válvulas. Aunque hubo muestras de estos tejidos y subgingivales, positivas para bacilos entéricos Gram negativos, no es posible asegurar que tuvieran el mismo origen filogenético.

Abstract Introduction: Periodontitis is an infectious disease that affects the support tissue of the teeth and it is associated with different systemic diseases, including cardiovascular disease. Microbiological studies facilitate the detection of microorganisms from subgingival and cardiovascular samples. Objective: To describe the cultivable periodontal microbiota and the presence of microorganisms in heart valves from patients undergoing valve replacement surgery in a clinic in Cali. Materials and methods: We analyzed 30 subgingival and valvular tissue samples by means of twophase culture medium, supplemented blood agar and trypticase soy agar with antibiotics. Conventional PCR was performed on samples of valve tissue. Results: The periodontal pathogens isolated from periodontal pockets were: Fusobacterium nucleatum (50%), Prevotella intermedia/ nigrescens (40%), Campylobacter rectus (40%), Eikenella corrodens (36.7%), Gram negative enteric bacilli (36.7%), Porphyromonas gingivalis (33.3%), and Eubacterium spp. (33.3%). The pathogens isolated from the aortic valve were Propionibacterium acnes (12%), Gram negative enteric bacilli (8%), Bacteroides merdae (4%), and Clostridium bifermentans (4%), and from the mitral valve we isolated P. acnes and Clostridium beijerinckii. Conventional PCR did not return positive results for oral pathogens and bacterial DNA was detected only in two samples. Conclusions: Periodontal microbiota of patients undergoing surgery for heart valve replacement consisted of species of Gram-negative bacteria that have been associated with infections in extraoral tissues. However, there is no evidence of the presence of periodontal pathogens in valve tissue, because even though there were valve and subgingival samples positive for Gram-negative enteric bacilli, it is not possible to maintain they corresponded to the same phylogenetic origin.