Incidencia y subregistro de casos de leptospirosis diagnosticados con tres métodos diferentes en Urabá, Colombia / Incidence and underreporting of leptospirosis comparing three diagnostic methods in the endemic region of Urabá, Colombia

Resumen Introducción. La leptospirosis representa un problema de salud pública y es una causa importante de morbimortalidad en la región de Urabá, cuya notificación se ve afectada por las deficiencias en el diagnóstico. Objetivo. Establecer la incidencia de la leptospirosis en los municipios del llamado 'eje bananero' de la región de Urabá, documentar la magnitud del subregistro y proponer orientaciones para el diagnóstico por laboratorio por parte de la red de salud pública. Materiales y métodos. Se compararon dos fuentes de información sobre la leptospirosis: el sistema oficial nacional de vigilancia y un estudio transversal de 479 pacientes febriles, llevado a cabo entre abril de 2010 y mayo de 2012. El diagnóstico se hizo con base en tres pruebas: inmunofluorescencia indirecta, microaglutinación y hemocultivo. La exhaustividad de cada fuente de información se estimó mediante el método de captura y recaptura. Resultados. El 58 % (278/479) de los pacientes fueron positivos para leptospirosis, por lo menos, en una de las pruebas y, el 10,43 % (29/278), en las tres. La inclusión de una cepa nativa en el panel de la prueba de microaglutinación aumentó el porcentaje de positividad en 15 %. La tasa acumulada de incidencia fue de 66,5 por 100.000 habitantes y la proporción de letalidad fue de 2,15 %. El subregistro de la morbilidad por leptospirosis en la región de Urabá, fue de 27,8 % y, el de la mortalidad, de 66,6 %. Conclusión. El subregistro de leptospirosis en la región reitera la necesidad de usar más de una prueba diagnóstica para identificar Leptospira spp. en pacientes de zonas endémicas. Este subregistro podría ser una situación común en todo el país.

Abstract Introduction: Leptospirosis represents a public health problem and is a significant cause of morbidity and mortality in the region of Urabá. However, its notification reveals diagnostic limitations. Objective: To establish the incidence of leptospirosis in the municipalities of the so-called eje bananero in the Urabá region, to describe the magnitude of underreporting, and to propose guidelines for laboratory diagnosis by the public health network. Materials and methods: Two leptospirosis information sources were used: The national official surveillance system and a cross-sectional study of 479 acute-phase patients from April, 2010, to May, 2012. The diagnosis was made using three different tests: Indirect immunofluorescence, microagglutination test, and blood cultures. The exhaustiveness percentage of each information source was calculatedusing thecapture and recapture test. Results: From the total number of cases, 58% (278/479) were positive for leptospirosis at least by a test and 10.43% (29/278) of cases were positive by all three methods. The inclusion of a native strain in the microagglutination test panel increased the percentage of positivity by 15%. The cumulative incidence rate was 66.5/100,000 inhabitants and the case fatality ratio was 2.15%. The underreporting rates of leptospirosis in the Urabá region were 27.8% in morbidity and 66.6% in mortality. Conclusion: Under-registration of leptospirosis in the region highlights the necessity to use more than one diagnostic test to identify Leptospira in patients from endemic areas. Under-registration could be a common situation throughout the country.

Detection of the mecA gene and identification of Staphylococcus directly from blood culture bottles by multiplex polymerase chain reaction

ABSTRACT Introduction: Staphylococcus spp. - both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) - are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality. Material and methods: In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive microorganisms confirmed by Gram-stain were analyzed. Results from multiplex PCR were compared to phenotypic characterization of isolates. Results: Staphylococcus aureus was detected in 85 (23.0%) blood cultures and CoNS in 286 (77.0%). There was 100% agreement between phenotypic and multiplex PCR identification. Forty-three (50.6%) of the 85 S. aureus carried the mecA gene and among the 286 CoNS, 225 (78.7%) were positive for the mecA gene. Conclusions: The multiplex PCR assay developed here was found to be sensitive, specific, rapid, and showed good agreement with the phenotypic results besides being less expensive. This PCR method could be used in clinical laboratories for rapid identification and initiation of specific and effective treatment, reducing patient mortality and morbidity. Furthermore, this method may reduce misuse of antimicrobial classes that are more expensive and toxic, thus contributing to the selection of antibiotic-resistant Staphylococcus spp.

Prevalencia de Bartonella henselae en donantes de sangre y riesgo de transmisión sanguínea en Chile / Prevalence of Bartonella henselae in blood donors and risk of blood transmission in Chile

Resumen Introducción: Bartonella henselae es el agente causal de la enfermedad del arañazo del gato en personas inmunocompetentes y de la angiomatosis bacilar y peliosis hepatis en inmunocomprometidos. En Chile la prevalencia de anticuerpos contra B. henselae en niños y adolescentes sanos es de 13,3%, en personas con riesgo ocupacional 60,5% y en gatos 85,6%. No existen datos publicados respecto de la seroprevalencia en donantes de sangre en nuestro país, por lo que determinar si B. henselae se encuentra presente en la sangre de los donantes al momento de la donación es muy importante, ya que este microorganismo puede sobrevivir hasta 35 días en los eritrocitos almacenados en banco de sangre a 4 °C. Objetivo: Determinar la prevalencia de B. henselae en donantes de sangre. Metodología: Se analizaron 140 muestras de sangre de donantes, para detectar la presencia de B. henselae, utilizando la técnica de la reacción de polimerasa en cadena (RPC). Resultados: Se obtuvo 13,6% de los donantes de sangre con RPC positiva para la B. henselae. La secuencia de los fragmentos amplificados presentó una identidad por sobre 98% con respecto a secuencias de B. henselae de referencia. Conclusión: El riesgo de transmisión sanguínea debiera ser considerado en un país con alta seroprevalencia de infección por B. henselae.

Background: Bartonella henselae is the causal agent of cat scratch disease in immunocompetent persons and bacterial angiomatosis in immunocompromised patients. In Chile, the prevalence of antibodies against B. henselae in healthy children and adolescents is 13.3%, in persons with occupational risk 60.5%, and in cats 85.6%. There are no published data regarding the seroprevalence in blood donors in our country, so determining if B. henselae is present in the blood of donors at the time of donation is very important, since this microorganism can survive up to 35 days in the red blood cells stored in a blood bank at 4 °C. Objective: To determine the prevalence of B. henselae in blood donors. Methodology: 140 donor blood samples were analyzed to detect the presence of B. henselae, using the polymerase chain reaction technique. Results: 13.6% of the blood donors with positive polymerase chain reaction for B. henselae were obtained. The sequence of the amplified fragments showed an identity of over 98% with respect to B. henselae reference sequences. Conclusion: The risk of blood transmission is due to a country with high B. henselae infection.

Direct identification from positive blood broth culture by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS)

ABSTRACT Bloodstream infections (BSIs) are among the most concerning bacterial infections. They are one of the leading causes of morbidity and mortality, and occur in 30-70% of critical care patients. The prompt identification of the causative microorganism can help choosing the appropriate antimicrobial therapy that will lead to better clinical outcomes. Blood culture is one of the most relevant tests for microbiological diagnosis of bacterial infections. The introduction of the MALDI-TOF microbiological diagnosis significantly decreased the time of identifying microorganisms. However, it depends on the growth on solid culture medium. In this study, 538 bottles of positive blood cultures were evaluated to test the accuracy of an in house modified protocol. The study sample consisted of 198 Gram-negative and 350 Gram-positive bacteria. In all, 460 (83.94%) species were identified based on the direct plate findings. The protocol allowed the identification of 185/198 (93.43%) of the Gram-negative bacteria, including aerobes, anaerobes, and non-fermenters, and 275/350 (78.85%) of the Gram-positive bacteria. The proposed method has the potential to provide accurate results in comparison to the traditional method with the potential to reduce the turnaround time for the results and optimize antimicrobial therapy in BSI.

Central-line-associated bloodstream infections in a resource-limited South African neonatal intensive care unit

Background. The rate of central-line-associated bloodstream infection (CLABSI) in South African (SA) public sector neonatal intensive care units (NICUs) is unknown. Tygerberg Children's Hospital (TCH), Cape Town, introduced a neonatal CLABSI surveillance and prevention programme in August 2012.Objectives. To describe CLABSI events and identify risk factors for development of CLABSI in a resource-limited NICU.Methods. A retrospective case-control study was conducted using prospectively collected NICU CLABSI events matched to four randomly selected controls, sampled from the NICU registry between 9 August 2012 and 31 July 2014. Clinical data and laboratory records were reviewed to identify possible risk factors, using stepwise forward logistic regression analysis.Results. A total of 706 central lines were inserted in 530 neonates during the study period. Nineteen CLABSI events were identified, with a CLABSI rate of 5.9/1 000 line days. CLABSI patients were of lower gestational age (28 v. 34 weeks; p=0.003), lower median birth weight (1 170 g v. 1 975 g; p=0.014), had longer catheter dwell times (>4 days) (odds ratio (OR) 5.1 (95% confidence interval (CI) 1.0 - 25.4); p=0.04) and were more likely to have had surgery during their NICU stay (OR 3.5 (95% CI 1.26 - 10); p=0.01). Significant risk factors for CLABSI were length of stay >30 days (OR 20.7 (95% CI 2.1 - 203.2); p=0.009) and central-line insertion in the operating theatre (OR 8.1 (95% CI 1.2 - 54.7); p=0.03). Gram-negative pathogens predominated (12/22; 54%), with most isolates (10/12; 83%) exhibiting multidrug resistance. Conclusion. The TCH NICU CLABSI rate is similar to that reported from resource-limited settings, but exceeds that of high-income countries. Prolonged NICU stay and central-line insertion in the operating theatre were important risk factors for CLABSI development. Intensified neonatal staff training regarding CLABSI maintenance bundle elements and hand hygiene are key to reducing CLABSI rates

Identification of Vaginal Fluid Using Microbial Signatures / 法医学杂志

OBJECTIVES@#To investigate the specific microbial signatures in vaginal fluid.@*METHODS@#Vaginal fluid （16 samples）, saliva （16 samples）, feces （16 samples）, semen （8 samples）, peripheral blood （8 samples）, urine （5 samples）, and nasal secretion （4 samples） were collected respectively. The 16S rRNA genes of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were amplified. PCR production was detected via a 3130xl Genetic Analyzer.@*RESULTS@#The detected number of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were 15, 5, 8, 14, and 3 in all vaginal fluid samples, respectively. Lactobacillus crispatus and Lactobacillus jensenii existed specifically in vaginal fluid.@*CONCLUSIONS@#There is a potential application value to detect Lactobacillus crispatus and Lactobacillus jensenii for the identification of vaginal fluid.

Utilización de espectrometría de masas para la identificación precoz de hemocultivos positivos: Validación de un algoritmo local / Use of mass spectrometry for rapid diagnosis of positive blood cultures: Validation of a local algorythm

Mass spectrometry (MS) is used in identification of positive blood culture, a contribution in the clinical management of septic patients. The protocol is labor intensive and disrupts the normal workflow in a clinical laboratory. We intended to make rapid diagnosis by using MS directly from shortly incubated blood agar plates (4 to 6 hours) comparing with results of the conventional method (MC). We worked in parallel 145 positive blood cultures, with correct identification by short method in 79% of cases. We observed better yield with non carbon bottles and with gramnegative rods. With this information we designed a rapid identification algorithm using MS, which allows advancing diagnosis in 12-16 hours, without increasing to the costs or work load, since extraction protocol is not used.

Espectrometría de masas (EM) es utilizada en identificación de hemocultivo positivo (HMP), constituyendo un aporte en el manejo clínico de paciente séptico. El protocolo de identificación directa es ultra laborioso, interrumpiendo el flujo de trabajo normal del laboratorio. Con el objetivo de hacer identificación rápida, utilizamos EM a partir de placas de agar sangre con incubación breve (4 a 6 h), comparando resultados con el método convencional en HMP. Se trabajó 145 frascos de HMP, identificando correctamente por método acortado 79% de los microorganismos. El rendimiento fue mejor en frascos sin carbón activado y en la identificación de bacilos gramnegativo. Con esta información, diseñamos algoritmo para la identificación precoz de hemocultivo positivo mediante EM, procesando a ciegas a las 4 a 6 h de incubación, lo que permite adelantar el diagnóstico en 12-16 h respecto del método tradicional, sin aumentar los costos ni la carga de trabajo, ya que no se utiliza protocolo de extracción.

Identificación rápida de patógenos en hemocultivos pediátricos por FilmArray / Rapid identification of pathogens from pediatric blood cultures by use of the FilmArray blood culture identification panel

Introduction: Dengue is a worldwide public health problem for which there is still no vaccine, so the knowledge of its temporal behavior could be useful for the implementation of appropriate control strategies. Objective: To analyze the incidence of dengue during the last ten years in Colombia (2004-2013), highlighting the periods and regions in which the largest number of cases was reported. Methods: We conducted a searching of dengue cases reported in Colombia between 2004 and 2013 in the database records of SIVIGILA and INS. The meteorological variables were obtained from IDEAM and its correlation with the incidence of dengue was found by the Pearson correlation method. Results: This analysis shows that every year there is an increase in the number of cases and that the most affected regions are the departments of Santander, Norte de Santander, Huila, Tolima and Valle del Cauca; with 2010 being the year with the highest record of events, with more than 150.000 cases. Discussion: The results indicate that DENV infection presents a cyclic behavior that most likely will be repeated every three or four years and the occurrence of cases can be attributed to social as well as climate changes.

Introducción: El dengue es un importante problema de salud pública mundial para el cual aún no existe una vacuna, por lo que el aporte al conocimiento de su comportamiento es útil para la ejecución de estrategias adecuadas para su control. Objetivo: Analizar la incidencia de dengue en los últimos diez años en Colombia (2004-2013), resaltando los períodos y regiones en las que se reporta el mayor número de casos. Métodos: Se realizó una búsqueda de casos de dengue reportados en Colombia entre 2004 y 2013 en registros del SIVIGILA y el INS. Las variables meteorológicas se obtuvieron del IDEAM y su correlación con la incidencia de dengue se halló por el método de correlación de Pearson. Resultados: Nuestro análisis muestra que cada año se presenta un incremento en el número de casos y las regiones más afectadas son Santander, Norte de Santander, Huila, Tolima y Valle del Cauca. El 2010 fue el año con mayor registro de eventos, con más de 150.000 casos. Discusión: Los resultados indican que la infección por DENV presenta un comportamiento cíclico, que muy posiblemente se repite cada tres o cuatro años y dicha ocurrencia de casos puede ser atribuida a cambios sociales y climáticos.

Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based VITEK MS System for the Identification of Acinetobacter Species from Blood Cultures: Comparison with VITEK 2 and MicroScan Systems

BACKGROUND: Acinetobacter species are the leading cause of bloodstream infection (BSI), but their correct identification is challenging. We evaluated the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based VITEK MS (bioMerieux, France), and two automated systems, VITEK 2 (bioMerieux) and MicroScan (Siemens, USA) for identification of Acinetobacter BSI isolates. METHODS: A total of 187 BSI isolates recovered at a university hospital in Korea between 2010 and 2012 were analyzed. The identification results obtained using VITEK MS and two automated systems were compared with those of rpoB sequencing. RESULTS: Of 187 isolates analyzed, 176 were identified to the species level by rpoB sequencing: the Acinetobacter baumannii group (ABG; 101 A. baumannii, 43 A. nosocomialis, 10 A. pittii isolates) was most commonly identified (82.4%), followed by Acinetobacter genomic species 13BJ/14TU (5.3%), A. ursingii (2.1%), A. soli (2.1%), A. bereziniae (1.1%), and A. junii (1.1%). Correct identification rates to the species group (ABG) level or the species level was comparable among the three systems (VITEK MS, 90.3%; VITEK 2, 89.2%; MicroScan, 86.9%). However, VITEK MS generated fewer misidentifications (0.6%) than VITEK 2 (10.8%) and MicroScan (13.1%) (P<0.001). In addition, VITEK MS demonstrated higher specificity (100%) for discrimination between ABG and non-ABG isolates than the other systems (both, 31.8%) (P<0.001). CONCLUSIONS: The VITEK MS system is superior to the VITEK 2 and MicroScan systems for identification of Acinetobacter BSI isolates, with fewer misidentifications and better discrimination between the ABG and non-ABG isolates.

Detection of Ehrlichia canis in domestic cats in the central-western region of Brazil

Ehrlichiosis is a worldwide distributed disease caused by different bacteria of the Ehrlichia genus that are transmitted by arthropod vectors. Its occurrence in dogs is considered endemic in several regions of Brazil. Regarding cats, however, few studies have been done and, consequently, there is not enough data available. In order to detect Ehrlichia spp. in cats from the central-western region of Brazil, blood and serum samples were collected from a regional population of 212 individuals originated from the cities of Cuiabá and Várzea Grande. These animals were tested by the Immunofluorescence Assay (IFA) and the Polymerase Chain Reaction (PCR) designed to amplify a 409 bp fragment of the dsb gene. The results obtained show that 88 (41.5%) cats were seropositive by IFA and 20 (9.4%) cats were positive by PCR. The partial DNA sequence obtained from PCR products yielded twenty samples that were found to match perfectly the Ehrlichia canis sequences deposited on GenBank. The natural transmission of Ehrlichia in cats has not been fully established. Furthermore, tick infestation was not observed in the evaluated cats and was not observed any association between age, gender and positivity of cats in both tests. The present study reports the first serological and molecular detection of E. canis in domestic cats located in the endemic area previously mentioned.

Relação entre a resistência a oxacilina e a produção de biofilme de amostras de origem comunitária e hospitalar pertencentes a diferentes espécies de Staphylococcus coagulase negativo / Relationship between resistance to oxacillin and biofilm production samples of Community and hospital belonging to different species of coagulase-negative Staphylococcus

Nas últimas décadas, os Staphylococcus coagulase-negativo, têm sido considerados como patógenos verdadeiros, sendo um dos principais grupos bacterianos responsáveis pelas infecções relacionadas a assistência a saúde (IRAS). O presente estudo teve como objetivo geral: avaliação da relação entre a resistência a oxacilina e a produção de biofilme de amostras Staphylococcus coagulase-negativo de origem comunitária e hospitalar. Neste sentido, foram desenvolvidos os seguintes objetivos específico: identificar ao nível de espécie os Staphylococcus coagulase-negativo; analisar por técnica fenotípica (Ágar vermelho do Congo) a produção de slime; avaliar quantitativamente, a produção de biofilme; correlacionar a produção de polissacarídeos extracelulares (slime) com a produção de biofilme; avaliar a relação da resistência a oxacilina como indicador da presença do gene mecA; avaliar a relação entre a concentração inibitória mínima e a concentração bactericida mínima para oxacilina; pesquisar a presença dos genes mecA, icaAD e atlE, pela técnica de PCR. Foi estudado um total de 150 amostras, sendo 50 isoladas de fômites, 50 isoladas de sangue e 50 isoladas de comunidade. Independente da origem, foram identificadas 14 espécies de Staphylococcus coagulase-negativo, sendo mais frequentes S. epidermidis 42,6%, S. haemolyticus 13,3% e S. cohnii cohnii 10,7%. A análise geral da expressão fenotípica de slime mostrou que 64% das amostras avaliadas eram produtoras de slime. Das 150 amostras testadas neste estudo, 95,3% foram produtoras de biofilme. Ao considerarmos a análise da quantificação do biofilme em relação às origens das amostras estudadas não encontramos diferenças significativas e a maioria das amostras foi considerada moderadamente produtora de biofilme. O gene mecA foi detectado em 6 amostras comunitárias, 34 amostras de fômites e 34 amostras de sangue. Não houve diferença significativa entre as amostras de fômites e sangue...

In recent decades, coagulase-negative Stapphylococci have been considered as true pathogen, one of the major bacterial groups responsible for hospital infection. The present study aimed to: assess the relationship between oxacillin resistance and biofilm production samples coagulase-negative Stapphylococci of community and hospital. In this sense, we have developed the following specific objectives: to identify to species level coagulase-negative Staphylococci; analyze by phenotypic test (Congo red Agar) slime production, evaluate quantitatively the biofilm production; correlate the production of extracellular polysaccharides (slime) with biofilm production; evaluate the relationship of resistance to oxacillin as an indicator of the presence of the mecA gene; evaluate the relationship between minimal inhibitory concentration and minimum bactericidal concentration for oxacillin; investigate the presence of the mecA gene, atlE and icaAD, by PCR. We studied a total of 150 samples, 50 were isolated from fomites, 50 from community and 50 isolated from blood. Regardless of origin, 14 species of coagulase-negative Stapphylococci were identified , being more frequent 42.6% S.epidermidis, 13.3% S. haemolyticus and 10.7% S. cohnii cohnii. A general analysis of the phenotypic expression of slime showed that 64% of the samples were slime producers...

Análisis de la monitorización de cinco indicadores de calidad del hemocultivo en un hospital universitario en Chile 2009-2011 / Quality indicators for blood culture: three years of monitoring at a university hospital in Chile

Blood culture is considered the "gold standard" for the diagnosis of bacteremia, critical condition with high morbidity and mortality. Because of its importance, it is estimated that the blood culture is a critical test that requires close monitoring on the quality with which the process is performed. The objective of this work is to show the results of the monitoring carried out during the past three years, of 5 quality indicators of blood cultures in the laboratory of the Hospital Clínico de la Pontificia Universidad Católica de Chile, considering pre-analytical, analytical and post-analytical aspects. In the 3 years monitored the mean contamination was 0,7%, 46% of adult bottles had adequate volume, match between Gram stain with final identification was 99.4%, 100% of correct participations were achieved in surveys of external quality control and Gram staining notification before 1 hour was 88.7%. With regard to proposed aims, in 2011 the laboratory complies with all, except the percentage of bottles with appropriate volume of blood inoculated. This indicator is very low and should be corrected as soon as possible since it is known that it is an important condition for optimum performance of blood cultures.

El hemocultivo es el "estándar de oro" para el diagnóstico de bacteriemia, condición grave de alta morbi-mortalidad. Por esto, se estima que el hemocultivo es un examen crítico, que requiere monitorización de su calidad. El objetivo de este trabajo es mostrar los resultados de la monitorización de 3 años de 5 indicadores de calidad del hemocultivo implementados en nuestro hospital. El porcentaje promedio de contaminación de las botellas para hemocultivo fue 0,7%, el porcentaje de botellas de adultos con volumen adecuado fue 46%, la concordancia de la tinción de Gram con la identificación final correspondió a 99,4%, el 100% de las participaciones en encuestas de control de calidad externo fueron correctas y 88,7% de los avisos de valores de alerta de la tinción de Gram fueron realizados antes de 1 hora. En el año 2011 se cumplió con las metas propuestas por el laboratorio para todos los indicadores, excepto con el porcentaje de botellas con volumen apropiado de sangre. Este último, se encuentra muy por debajo de la meta y debe ser mejorado a la brevedad, ya que el volumen de sangre cultivada es el factor más importante para obtener un rendimiento óptimo del hemocultivo.

Dos aislamientos de Salmonella Infantis multirresistentes se comportan como hipoinvasivos pero con elevada proliferación intracelular

En este trabajo se investigó la presencia de determinantes característicos de plásmidos de virulencia en dos aislamientos clínicos de Salmonella Infantis portadores de plásmidos de multirresistencia. Además, se estudió la capacidad de invasión y proliferación en células eucariotas no fagocíticas. Ninguno de los aislamientos de S. Infantis mostró los determinantes genéticos que caracterizan a los plásmidos de virulencia para este género (operón spv). Los ensayos de invasión sobre líneas celulares eucariotas mostraron que los aislamientos de S. Infantis presentan una capacidad de invasión disminuida pero persisten y proliferan en el citoplasma, independientemente de utilizar una línea celular permisiva (HeLa) o no permisiva (NRK) para tal fin. Finalmente, no se observaron indicios microscópicos que podrían hacer sospechar un efecto bactericida de estas líneas celulares sobre los aislamientos estudiados.

Two multidrug-resistant Salmonella Infantis isolates behave like hypo-invasive strains but have high intracellular proliferation. In this work, plasmid-encoded virulence factors in two Salmonella Infantis isolates carrying multiresistance plasmids were investigated. In addition, their invasion and proliferative ability in non-phagocytic cells was studied. None of them showed the typical determinants of virulence plasmids (spv operon). The invasion assays of S. Infantis isolates on eukaryotic cells showed a decreased ability to Invade but they remained and proliferated In the cytoplasm regardless of having used a permissive (HeLa) or non-permissive (NRK) cell line. Finally, there was no microscopic evidence suggesting a bactericidal effect of these eukaryotic cell lines on the Isolates tested.

Vertebro-cerebral cryptococcosis mimicking tuberculosis: A diagnostic dilemma in countries with high burden of tuberculosis.

We report a case of a 30-year-old immunocompetent man with disseminated cryptococcosis who was initially treated with antitubercular therapy due to clinical and radiological diagnosis of vertebro-cerebral tuberculosis. The diagnosis of Cryptococcus infection was made due to incidental isolation of this fungus from blood culture with negative cerebrospinal fluid culture results. Though disseminated cryptococcosis with central nervous system, skeletal, and skin involvement is an uncommon manifestation of Cryptococcus neoformans infection, a high clinical suspicion and early initiation of therapy is needed to recognise and treat such patients efficiently.

Detection of bacterial growth in blood components using oxygen consumption as a surrogate marker in a tertiary oncology setup.

Microbiological contamination of blood and blood products is a well-recognised transfusion risk. This study was performed in the blood bank of our oncology centre, with an objective to detect bacterial contamination in our blood products using oxygen consumption as a surrogate marker [Pall Enhanced Bacterial Detection System (eBDS)]. Results revealed that the percentages of failed units were 1.16% for random donor platelets (RDP), 0.81% for single donor platelets (SDP) and 2.94% for packed red blood cells (PRBCs), of which one RDP and one SDP grew coagulase-negative staphylococcus, while one PRBC culture grew Gram-positive bacilli.

Comparative evaluation of paired blood culture (aerobic/aerobic) and single blood culture, along with clinical importance in catheter versus peripheral line at a tertiary care hospital.

Purpose: Paired blood culture (PBC) is uncommon practice in hospitals in India, leading to delayed and inadequate diagnosis. Also contamination remains a critical determinant in hampering the definitive diagnosis. Objectives: To establish the need of PBC over single blood culture (SBC) along with the degree of contamination, this comparative retrospective study was initiated. Materials and Methods : We processed 2553 PBC and 4350 SBC in BacT/ALERT 3D (bioMerieux) between October 2010 and June 2011. The positive cultures were identified in VITEK 2 Compact (bioMerieux). True positivity and contaminants were also analyzed in 486 samples received from catheter and peripheral line. Results : Out of 2553 PBC samples, positivity was seen in 350 (13.70%). In 4350 SBC samples, positivity was seen in 200 samples (4.59%). In PBC true pathogens were 267 (10.45%) and contaminants were 83 (3.25%), whereas in SBC 153 (3.51%) were true positives and contaminants were 47 (1.08%). Most of the blood cultures (99.27 %) grew within 72 h and 95.8% were isolated within 48 h. In 486 PBCs received from catheter/periphery (one each), catheter positivity was found in 85 (true positives were 48, false positives 37). In peripheral samples true positives were 50 and false positives were 8. Conclusion: Significantly higher positive rates were seen in PBCs compared with SBCs. Automated blood culture and identification methods significantly reduced the time required for processing of samples and also facilitated yield of diverse/rare organisms. Blood culture from catheter line had higher false positives than peripheral blood culture. Thus every positive result from a catheter must be correlated with clinical findings and requires further confirmation.