The hpx genetic system for hypoxanthine assimilation as a nitrogen source in Klebsiella pneumoniae: gene organization and transcriptional regulation.

Growth experiments showed that adenine and hypoxanthine can be used as nitrogen sources by several strains of K. pneumoniae under aerobic conditions. The assimilation of all nitrogens from these purines indicates that the catabolic pathway is complete and proceeds past allantoin. Here we identify the genetic system responsible for the oxidation of hypoxanthine to allantoin in K. pneumoniae. The hpx cluster consists of seven genes, for which an organization in four transcriptional units, hpxDE, hpxR, hpxO and hpxPQT, is proposed. The proteins involved in the oxidation of hypoxanthine (HpxDE) or uric acid (HpxO) did not display any similarity to other reported enzymes known to catalyze these reactions, but instead are similar to oxygenases acting on aromatic compounds. Expression of the hpx system is activated by nitrogen limitation and by the presence of specific substrates, with hpxDE and hpxPQT controlled by both signals. Nitrogen control of hpxPQT transcription, which depends on 54, is mediated by the Ntr system. In contrast, neither NtrC nor NAC is involved in the nitrogen control of hpxDE, which is dependent on 70 for transcription. Activation of these operons by the specific substrates is also mediated by different effectors and regulatory proteins. Induction of hpxPQT requires uric acid formation, whereas expression of hpxDE is induced by the presence of hypoxanthine through the regulatory protein HpxR. This LysR-type regulator binds to a TCTGC-N4-GCAAA site in the intergenic hpxD-hpxR region. When bound to this site for hpxDE activation, HpxR negatively controls its own transcription.

Ultrasound scan as a potential source of nosocomial and cross-infection: a literature review

AbstractThe authors review the main concepts regarding the importance of cleaning/disinfection of ultrasonography probes, aiming a better comprehension by practitioners and thus enabling strategies to establish a safe practice without compromising the quality of the examination and the operator productivity. In the context of biosafety, it is imperative to assume that contact with blood or body fluids represents a potential source of infection. Thus, in order to implement cleaning/disinfection practice, it is necessary to understand the principles of infection control, to consider the cost/benefit ratio of the measures to be implemented, and most importantly, to comprehend that such measures will not only benefit the health professional and the patient, but the society as a whole.

Molecular epidemiology and extended-spectrum β-lactamases production of Klebsiella pneumoniae isolated from three dairy herds

The objectives of this study were to isolate Klebsiella pneumoniae from different sources in three dairy cattle herds, to use the pulsed-field gel electrophoresis (PFGE) to measure genotypic similarities between isolates within a dairy herd, to verify the production of extended-spectrum β-lactamases (ESBLs) by the double-disk synergy test (DDST), and to use the PCR to detect the main ESBLs subgroups genes. Three dairy farms were selected based on previous mastitis outbreaks caused by K. pneumoniae. Milk samples were collected from lactating cows and from the bulk tank. Swabs were performed in different locations, including milking parlors, waiting room, soil, animal's hind limbs and rectum. K. pneumoniae was isolated from 27 cases of intramammary infections (IMI) and from 41 swabs. For farm A isolates from IMI and bulk tank were considered of the same PGFE subtype. One isolate from a bulk tank, three from IMI cases and four from environmental samples were positive in the DDST test. All eight DDST positive isolates harbored the bla shv gene, one harbored the bla tem gene, and three harbored the bla ctx-m gene, including the bulk tank isolate. Our study confirms that ESBL producing bacteria is present in different locations in dairy farms, and may be responsible for IMI. The detection of ESBLs on dairy herds could be a major concern for both public and animal health.

Research of Klebsiella pneumoniae in dairy herds

Klebsiella pneumoniae is a common environmental agent of clinical and subclinical mastitis affecting dairy herds, and may be present in the final product decreasing its quality. Mastitis caused by K. pneumoniae is even more severe due to its poor response to antibiotic therapy, rapid evolution to toxic shock and death of the animal. This paper aimed to study the prevalence of this pathogen among dairy herds in ten farms located in different municipalities of São Paulo State based on size and use of milking technology. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20mL) was collected from all CMT-positive quarters and bulk tanks, whereas swab samples were collected from feces, hind limbs of the animals, bedding and milking parlor. Identification of K. pneumoniae was performed using conventional microbiology culture, biochemical assay and Polimerase Chain Reaction. The primers were designed and tested at the Laboratory of Molecular Biology applied to Zoonoses (FMVZ, Unesp-Botucatu) targeting the 16S rRNA gene. This study included 1067 animals. Six cases of intramammary infection by K. pneumoniae were detected in six different cows in two farms. Moreover, K. pneumoniae was isolated in 77 swabs (34 from bedding in 9 farms, 7 from waiting rooms in 5 farms, 6 from milking parlors in 4 farms, 11 from rectums in six farms, and 19 from hindlimbs in 7 farms. Molecular analysis confirmed the agent was K. pneumoniae. At least one strain of the agent was identified in a certain site in all farms, showing the need of maintaining the hygiene in dairy farms.

Use of molecular epidemiology to monitor the nosocomial dissemination of methicillin-resistant Staphylococcus aureus in a University Hospital from 1991 to 2001

Methicillin-resistant Staphylococcus aureus (MRSA) has been the cause of major outbreaks and epidemics among hospitalized patients, with high mortality and morbidity rates. We studied the genomic diversity of MRSA strains isolated from patients with nosocomial infection in a University Hospital from 1991 to 2001. The study consisted of two periods: period I, from 1991 to 1993 and period II from 1995 to 2001. DNA was typed by pulsed-field gel electrophoresis and the similarity among the MRSA strains was determined by cluster analysis. During period I, 73 strains presented five distinctive DNA profiles: A, B, C, D, and E. Profile A was the most frequent DNA pattern and was identified in 55 (75.3%) strains; three closely related and four possibly related profiles were also identified. During period II, 80 (68.8%) of 117 strains showed the same endemic profile A identified during period I, 18 (13.7%) closely related profiles and 18 (13.7%) possibly related profiles and, only one strain presented an unrelated profile. Cluster analysis showed a 96% coefficient of similarity between profile A from period I and profile A from period II, which were considered to be from the same clone. The molecular monitoring of MRSA strains permitted the determination of the clonal dissemination and the maintenance of a dominant endemic strain during a 10-year period and the presence of closely and possibly related patterns for endemic profile A. However, further studies are necessary to improve the understanding of the dissemination of the endemic profile in this hospital.

Análisis de algunas características físicas y químicas de un biopolímero producido por Klebsiella pneumoniae en un proceso de fermentación simple

Tesis (Maestría en Ciencias con Especialidad en Ingeniería Cerámica) U.A.N.L.

Factores de riesgo para infecciòn o colonizacion por Klebsiella pneumoniae resistente a carbapenemicos en un Hospital Universitario de Tercer Nivel en Bogota. 2009-2010

Introducción: La prevalencia de infecciones intrahospitalarias por Klebsiella pneumoniae resistente a carbapenémicos, se ha incrementado en la población hospitalizada constituyendo un problema de gran magnitud por su alta morbilidad y mortalidad. Objetivo: Identificar los factores de riesgo asociados a infección o colonización por Klebsiella pneumoniae resistente a carbapenémicos en pacientes hospitalizados. Metodología: Estudio de casos y controles pareado por tipo de muestra microbiológica de enero 2009 a Abril 2011. Casos: pacientes con diagnóstico de infección o colonización por Klebsiella pneumoniae resistente a carbapenémicos. Controles: pacientes hospitalizados en el mismo periodo de los casos con infección o colonización por Klebsiella pneumoniae sensible a carbapenémicos. Muestra 99 pacientes. 33 casos y 66 controles. Resultados: Se confirmó la presencia de un brote de infección por Klebsiella pneumoniae resistente a carbapenémicos tipo KPC3, el análisis bivariado demostró factores de riesgo asociados: El uso previo de antibióticos (p 0.004), particularmente cefepime (p 0.021) y carbapenem (p 0.019) , los dìas de ventilación mecánica (p 0.003), los días de uso de catéter central (p 0.016), los días de estancia en UCI antes del aislamiento (p 0.003) y el tiempo de estancia hospitalaria total antes del aislamiento (p 0.001), el análisis multivariado se encontró una asociación significativa en el número de días de uso previo de carbapenémicos OR de 2.08 (IC 1.03 – 4.17) (p 0.04) . La mortalidad atribuible fue del 25%. Conclusión: Los días de uso previo de carbapenémicos se relacionan con la infección o colonización por Klebsiella pneumoniae resistente a carbapenémicos.

Genotyping Klebsiella pneumoniae isolated from hepatic abscesses in three patients from Bogota, Colombia

Pyogenic liver abscess caused by Klebsiella pneumoniae represents an ever increasing entity which has mainly been described as occurring in Asia, even though, on a smaller scale, cases are being more frequently described from the USA and Europe, 13% overall mortality being reached worldwide. Affected patients are severely sick, suffering from fever, sweating, having increased acute phase reactants and risk factors such as Diabetes Mellitus, alcoholism and the inherent characteristics of the bacteria causing the disease. Objective: in this work we used a Multilocus Sequencing Typing (MLST), a nucleotide sequence-based method in order to characterize the genetic relationships among bacterial isolates. Materials and methods: the report is focused on three cases involving patients suffering from pyogenic liver abscess caused by Klebsiella pneumoniae in two hospitals in Bogota, Colombia, where phenotyping and hypermucoviscosity studies were carried out, as well as the genotyping of cultured Klebsiella isolates. Reults: it was found that the isolated microorganism in cases I and II corresponded to the same K. pneumoniae strain, having 100% sequence identity for the 5 genes being studied while the strain in Case III was genotypically different. Conclusion: it is important to carry out multidisciplinary studies allowing all pyogenic liver abscess cases reported in Colombia to be complied to ascertain the frequency of microorganisms causing this pathology in our country, as well as a genotyping study of different K. pneumoniae strains to compare them and confirm clonal and pathogenicity relationships through housekeeping gene analysis.

Construcción de una filogenia molecular para las especies de los géneros Klebsiella y Raoultella basada en los genes ARNr 16S y ARN polimerasa subunidad

Las bacterias de los géneros Raoultella y Klebsiella son patógenos oportunistas para las cuales no existe un sistema uniforme de clasificación taxonómica internacional. En el presente estudio se propone una filogenia molecular basada en el gen ribosomal 16S (ADNr 16S) y el gen codificante de la subunidad de la ARN polimerasa (rpoB) de los géneros Klebsiella y Raoultella con el fin de establecer relaciones evolutivas entre dichos géneros. Los resultados evidencian una agrupación acorde con la taxonomía y las propiedades bioquímicas características, reportadas en el Genbank. Se estableció una bifurcación en los árboles, lo cual confirma la separación de los géneros Klebsiella y Raoultella. Adicionalmente, se confirmó el carácter polifilético de K. aerogenes por el gen ADNr 16S y la agrupación de R. terrigena y K. oxytoca de acuerdo con el gen rpoB. La comparación entre los árboles obtenidos permitió determinar relaciones evolutivas entre las especies, a partir de los genes evaluados, lo cual refleja cambios aparentes a nivel taxonómico y corrobora la importancia del análisis a nivel de multilocus. Este tipo de estudios permite monitorear la estabilidad de los genotipos microbianos sobre la escala temporal y espacial, mejorar la precisión de las anotaciones taxonómicas (mejor descripción de taxones o subdivisiones genéticas) y evaluar la diversidad genética y adaptabilidad en términos de virulencia o resistenciaa drogas.

Prevalencia de betalactamasas de espectro extendido en Escherichia coli, Klebsiella pneumoniae y Klebsiella oxytoca del Hospital Occidente de Kennedy. Nivel III, Bogotá

El objetivo es calcular la prevalencia para Escherichia coli, Klebsiella pneumoniae y Klebsiella oxytoca, productoras de betalactamasas espectro extendido, en el Hospital Occidente de Kennedy Nivel III, de Bogotá. Metodología: se analizaron en el Hospital Occidente de Kennedy, durante el período comprendido entre el 20 de noviembre de 2002 y el 30 septiembre de 2003, 3.574 cultivos, en los cuales se identificaron 897 cepas de Kepsiella pneumoniae, Klebsiella oxytoca y Escherichia coli, mediante paneles de microdilución del sistema de MicroScan Dried Gram Negative, como prueba de cribado para la identificación de germen y susceptibilidad a betalactamasas de espectro extendido. Luego se realizó una prueba confirmatoria con paneles de sistema MicroScan Dried ESBL Confirmation, recomendada para su uso por el National Committee for Clinical Laboratory Standards (NCCLS), al evaluar la concentración inhibitoria mínima para ceftazidima y cefotaxima solos y en combinación con ácido clavulánico. Resultados: los resultados mostraron una prevalencia combinada de gérmenes productores betalactamasas igual a 18,6% (intervalo de confianza del 95%: 16,2%-21,4%). La prevalencia para Escherichia coli fue de 9,5%; para Klebsiella pneumoniae, de 43,5%, y para Klebsiella oxytoca, de 10,3%. Los índices de resistencia bacteriana más altos correspondieron a ceftriaxona y ceftazidima. Conclusión: el estudio demuestra una alta prevalencia de betalactamasas de espectro extendido en gérmenes gramnegativos, probablemente por el uso excesivo de antibióticos betalactámicos de amplio espectro. Además, se destaca la importancia de la detección con pruebas de susceptibilidad y confirmación, como apoyo para la instauración de medidas de control y de vigilancia epidemiológica, con el fin de reducir índices de resistencia bacteriana emergente.

Evaluación muticéntrica de la actividad antimicrobiana in vitro de tigeciclina en aislados obtenidos de pacientes hospitalizados en Colombia

El aumento en la resistencia bacteriana es un problema global que afecta también a nuestro país y que puede llevar a fracasos terapéuticos, peores desenlaces clínicos, uso de terapia combinada, mayor riesgo de interacciones medicamentosas y de reacciones adversas asi como a aumento de los costos del tratamiento. Tigeciclina, un nuevo antibiótico de la clase de las glicilciclinas ha mostrado una promisoria actividad antibacteriana in vitro contra bacterias comunes incluyendo multiresistentes que cada vez son más prevalentes en nuestro país especialmente a nivel hospitalario. Este estudio fue desarrollado para determinar la actividad de tigeciclina en nuestro medio y conocer su potencial utilidad.

Resultados de la campaña para el control de infección nosocomial en la UCI. Clínica San Rafael 2006 - 2007

Las entidades de salud de alta complejidad, atienden pacientes generalmente en condición crítica, requiriendo intervenciones y procedimientos especializados con uso frecuente de las unidades de cuidado intensivo, dado que su condición clínica puede estar seriamente afectada y su pronóstico no siempre ser satisfactorio, generando a la vez altos costos en la atención y prolongación de las estancias. La infección nosocomial constituye un factor relevante que afecta frecuentemente la evolución y el pronóstico de los pacientes en cuidado crítico, que afrontan situaciones como eventos de inmunosupresión, procedimientos invasivos diagnósticos o terapéuticos que los predisponen a adquirir infecciones dentro del hospital.