Effect of daily consumption of extra virgin olive oil on the lipid profile and microbiota of HIV-infected patients over 50 years of age

Extra virgin olive oil (EVOO) has shown beneficial effects on the lipid profile and inflammatory parameters in general population. Our goal is to analyze these changes together with those of intestinal microbiota in human immunodeficiency virus (HIV)-infected patients over 50 years of age. Total cholesterol decreased significantly (5 mg/dL), and a nonsignificant decrease in low-density lipoprotein cholesterol (12 mg/dL), triglycerides (21 mg/dL), and CRP (1.25 mg/dL) was observed. There was a significant increase in alpha diversity after the intervention in men and a decrease in proinflammatory genera such as Dethiosulfovibrionaceae was observed. Differences were also observed in the microbiota of men and women and according to the type of antiretroviral treatment. Sustained consumption of 50g of EVOO in elderly HIV-infected patients might be associated with an improvement in lipid profile and alfa diversity of intestinal microbiota.This work was financed in part by Grants from Plan Nacional de I+D+I and Fondo Europeo de Desarrollo Regional-FEDER (RD16/0025/0040; http:// www.isciii.es/isciii/es/contenidos/fd-investigacion/fd-ejecucion/fd-centrosparticipados/ centros-participados-redesretics.shtml) and Fondo de Investigación Sanitaria (PI 18/00819)

Aerococcus urinae: a rare pathogen in urinary tract infections, associated with patients with underlying urinary pathology

Introducción: Aerococcus urinae es un patógeno urinario poco frecuente que ha sido asociado en la bibliografía a pacientes en la tercera edad, con patología urinaria subyacente. La mala identificación mediante pruebas bioquímicas convencionales, su baja tasa de aislamiento y la similar morfología con otros microorganismos considerados como flora normal en el tracto urinario, hacen de este microorganismo un gran desconocido. El objetivo de este trabajo fue determinar la implicación clínica de Aerococcus urinae en pacientes con infección urinaria, estudiando la relación existente descrita en la bibliografía con pacientes en la tercera edad, con patología urinaria subyacente. Asimismo, debido a su infradiagnóstico en infección urinaria (ITU), se evaluó la utilidad de la espectrometría de masas MALDI-TOF (Bruker) como modelo diagnóstico confiable en el laboratorio de microbiología. Material y métodos: Se realizó un estudio prospectivo en pacientes con infección urinaria desde mayo a septiembre de 2014. Se estudiaron los perfiles de sensibilidad a los antimicrobianos frecuentemente utilizados en infección urinaria. Se registraron los datos clínicos más importantes de los pacientes con un recuento positivo para A. urinae. Resultados: De las 9261 muestras de orina analizadas, 1513 muestras resultaron tener un recuento mayor a 100.000 UFC/mL. A. urinae fue aislado en 3 casos, y la identificación por MALDI-TOF fue fiable a nivel de género y especie ( score ≥ 2) , siendo contrastada mediante secuenciación ARNr 16S. Todos los pacientes que desarrollaron infección urinaria por A. urinae, fueron pacientes en la tercera edad con una patología subyacente, siendo en todos los casos este microorganismo resistente a trimetropim/sulfametoxazol. Conclusiones: La implicación de A. urinae como patógeno urinario, en pacientes con patología urinaria de base, la dificultad en su diagnóstico, y la alta tasa de resistencia de este microorganismo a trimetropim/sulfametoxazol, hacen recomendable establecer una especial atención en los métodos diagnósticos utilizados.Introduction: Aerococcus urinae is an uncommon urinary tract pathogen that has been associated in the literature for older patients with underlying urinary pathology. Misidentification by conventional biochemical tests, their low rate of isolation and similar morphology to other microorganisms considered as normal flora in the urinary tract, makes this organism a great unknown. The aim of this investigation is to determine the implication of Aerococcus urinae as the cause of urinary tract infections, studying the relationship described in literature in elderly patients with underlying urinary pathology. Material and methods: A prospective study on patients with urinary infection, from May to September 2014 was performed. Urine-cultures with a significant bacteria and suspicious of A. urinae, were identified by means of MALDI-TOF system, and contrasted with ARNr 16S sequencing. Profiles of antimicrobial susceptibility frequently used in urinary tract infection were studied. Clinical data for patients with positive urine cultures for A. urinae were registered. Result: 9261 urine samples were analyzed, 1513 samples had counts greater than 100,000 CFU / mL. A. urinae was isolated in 3 cases, and identification by MALDI-TOF was reliable genus and species level (score ≥ 2), being proven by sequencing 16S rRNA. All patients who developed urinary infection by A. urinae were elderly patients with underlying pathology, and this microorganism resistant to trimethoprim / sulfamethoxazole. Conclusions: The A. urinae overt implication as urinary pathogen, the high rate of resistance of this organism to trimethoprim / sulfamethoxazole, and its difficulty to diagnose, urge to pay special attention to the diagnostic methods applied

Carbapenemase detection in Pseudomonas aeruginosa by MALDI-TOF MS mass spectrometry

Introducción: Las bacterias gramnegativas, especialmente Pseudomonas, presentan con frecuencia resistencia a múltiples antibióticos incluyendo carbapenemes. La resistencia a carbapanemes se ha incrementado en los últimos años causada por alteraciones de membrana o por la producción de carbapenemasas. Objetivo: Valorar la utilización de la espectrometría de masas MALDI-TOF MS® para la detección de carbapenemasas clase A o B en Pseudomonas aeruginosa. Material y métodos: Partiendo de 12 aislados de Pseudomonas aeruginosa productoras de carbapenemasas clase A o B identificados mediante método de difusión disco-placa, tipificadas usando los discos: meropenem 10µg, meropenem 10µg + ácido borónico, meropenem 10µg + cloxacilina y meropenem 10µg + ácido dipicolínico (Rosco Diagnostica), hemos analizado posibles picos de hidrólisis del meropenem tras la acción de las carbapenemasas mediante espectrometría de masas MALDI-TOF MS®. Como controles negativos se utilizaron 25 cepas de Pseudomonas aeruginosa sensibles a meropenem y 8 cepas de Pseudomonas aeruginosa con impermeabilidad de membrana, no detectables mediante la metodología utilizada. Resultados: De las 12 cepas productoras de carbapenemasas clase A o B, (2/12 clase A, 10/12 clase B), la técnica de espectrometría de masas MALDI-TOF MS® detectó picos de degradación del antibiótico en estudio correspondientes a la presencia de carbapenemasas en 11/12 casos (94.4%). En las cepas usadas como controles negativos, la espectrometría de masas MALDI-TOF MS® indicó la ausencia de carbapenemasas clase A o B en 31/33 (93.9%) casos. Conclusión: La espectrometría de masas MALDI-TOF MS® puede ser una herramienta útil para la confirmación de carbapenemasas clase A y B en Pseudomonas aeruginosa.Introduction: Gram-negative bacteria especially Pseudomonas are resistance to multiple antibiotics including carbapenems. Carbapanemes resistance has increased in recent years caused by alterations of membrane or the production of carbapenemases. Objective: Assess the use of MALDI-TOF MS® mass spectrometry for the detection of carbapenemases class A or B in Pseudomonas aeruginosa. Material and methods: From isolated from Pseudomonas aeruginosa producing carbapenemases 12 class A or B identified by diffusion method disco-plate, classified using disks: meropenem 10μg, meropenem 10μg + boronic acid, meropenem 10μg + cloxacillin and meropenem 10μg + acid dipicolinic (Rosco Diagnostica), we analyzed possible hydrolysis of meropenem peaks after the action of the carbapenemases by MALDI-TOF MS® mass spectrometry. As negative controls were used 25 strains of Pseudomonas aeruginosa sensitive to meropenem and 8 strains of Pseudomonas aeruginosa with waterproof membrane, not detectable by the methodology used. Results: Of the 12 strains producing carbapenemases class A or B, (2/12 class A, 10/12 class B), MALDI-TOF MS® mass spectrometry technique detected peaks of degradation of the antibiotic in study to the presence of carbapenemases in 11/12 cases (94.4%). The strains used as controls negative, MALDI-TOF MS® mass spectrometry indicated the absence of carbapenemases class A or B at 31/33 cases (93.9%). Conclusion: MALDI-TOF MS® mass spectrometry can be a useful tool for the confirmation of carbapenemases class A and B in Pseudomonas aeruginosa

Pyrosequencing Analysis Reveals Changes in Intestinal Microbiota of Healthy Adults Who Received a Daily Dose of Immunomodulatory Probiotic Strains

The colon microbiota plays a crucial role in human gastrointestinal health. Current attempts to manipulate the colon microbiota composition are aimed at finding remedies for various diseases. We have recently described the immunomodulatory effects of three probiotic strains (Lactobacillus rhamnosus CNCM I-4036, Lactobacillus paracasei CNCM I-4034, and Bifidobacterium breve CNCM I-4035). The goal of the present study was to analyze the compositions of the fecal microbiota of healthy adults who received one of these strains using high-throughput 16S ribosomal RNA gene sequencing. Bacteroides was the most abundant genus in the groups that received L. rhamnosus CNCM I-4036 or L. paracasei CNCM I-4034. The Shannon indices were significantly increased in these two groups. Our results also revealed a significant increase in the Lactobacillus genus after the intervention with L. rhamnosus CNCM I-4036. The initially different colon microbiota became homogeneous in the subjects who received L. rhamnosus CNCM I-4036. While some orders that were initially present disappeared after the administration of L. rhamnosus CNCM I-4036, other orders, such as Sphingobacteriales, Nitrospirales, Desulfobacterales, Thiotrichales, and Synergistetes, were detected after the intervention. In summary, our results show that the intake of these three bacterial strains induced changes in the colon microbiota.This study utilized fecal samples from the clinical trial NCT01479543 that was supported by Hero Spain S. A. through contract #3582 with the Fundacion General Empresa Universidad de Granada. Carolina Gomez-Llorente is the recipient of a postdoctoral fellowship of the University of Granada

Sepsityper® for rapid identification of microorganisms from positive blood cultures

Nuestro estudio ha evaluado las ventajas de la utilización del kit Sepsityper® para la identificación rápida de microorganismos a partir de hemocultivos positivos, acompañado de la tecnología de espectrometría de masas MALDITOF MS®, en comparación con los métodos tradicionales empleados para el diagnóstico de bacteriemia. Para la identificación del microorganismo en 379 hemocultivos positivos en el Departamento de Microbiología del Hospital Universitario San Cecilio, se aplicó la espectrometría de masas MALDITOF MS® utilizando el sistema Sepsityper® (Bruker) y se comparó con la identificación mediante métodos convencionales (Wider, Vitek II, Api). La correlación de resultados de los dos esquemas diagnósticos fue determinada estadísticamente por el coeficiente de correlación kappa. La distribución de los aislamientos fue de un 24,7 % de Bacilos Gram negativos (BGN) y 75,3 % de microorganismos Cocos Gram positivos (CGP). La concordancia global de resultados fue del 95,8 % en la especie (k = 0,928) y del 98,7 % en el género (k = 0,977), siendo el porcentaje de identificaciones fallidas del 1,3%. Para BGN hubo una concordancia de resultados del 95,2 % (k = 0,928, especie), y 100 % (k = 1, género). Respecto a los CGP, la concordancia fue del 98,2 % en género (k = 0,931), y del 82,5 % (k = 0,627) a nivel de especie. En nuestra experiencia se ha observado una ganancia de al menos 13–23h en la identificación a nivel de especie. La utilización del kit Sepsityper® para la identificación rápida de microorganismos a partir de hemocultivos positivos, acompañado de MALDITOF MS®, muestra una excelente correlación respecto a la identificación realizada a través de la metodología convencional, con una importante disminución del tiempo hasta la identificación.Our study has evaluated the advantages of using Sepsityper® kit for a fast identification of microorganisms from positive blood cultures, along with the mass spectrometry technology MALDITOF-MS®, compared to traditional methods used for diagnosis of bacteremia. To identify the microorganism isolated from the 379 positive blood cultures (BC +) the Department of Microbiology, University Hospital San Cecilio, MALDITOFMS® mass spectrometry along with Sepsityper® (Bruker) were applied and it was compared to the conventional methods for the identification of this organism. The correlation of results between these two diagnostic schemes was statistically determined by kappa correlation coefficient. The distribution of the isolates was 24.7% for Gram negative bacilli (GNB) and 75.3% for Gram-positive cocci (GPC). The overall concordance of results was 95.8% within the species (k = 0.928) and 98.7% within the genus (k = 0.977), with a failed-identification percentage of 1.3%. For GNB there was a concordance of results of 95.2% (k = 0.928, species), and 100% (k = 1, genus). Regarding the GPC, the concordance was 98.2% within the genus (k = 0.931), and 82.5% (k = 0.627) at the species level. According to our experience there was a gain of at least 13-23 hours in the identification of the microorganisms at the species level. The use of Sepsityper® kit for the rapid identification of microorganisms from positive blood cultures, along with MALDITOF-MS®, show an excellent correlation compared to identification made by the conventional methods, with a significant reduction in time until identification

Evaluación de los métodos de genotipado del virus de la hepatitis C y detección del polimorfirmo Q80K del GEN NS3 para pacientes naïve a los nuevos antivirales de acción directa

Premio a la mejor comunicación del XVI Congreso de la Sociedad Andaluza de Enfermedades InfecciosasDesde el año 2012 en que se aprueban los primeros Antivirales de Acción Directa (AADs), el tratamiento de la hepatitis C (VHC) ha experimentado un cambio histórico y se está viviendo un momento muy esperanzador para erradicar esta enfermedad. Los resultados de los ensayos clínicos realizados con estos nuevos tratamientos alcanzan tasas de respuestas virales del 90 al 100% dependiendo de la combinación del fármaco y del genotipo del virus de la hepatitis C (VHC). Y esto es así, las guías nacionales e internacionales (AASLD/IDSA., 2015) (EASL, 2015) (GEHEP 2015) nos van a recomendar emplear una u otra combinación de AADs según la condición clínica del paciente (en especial grado de fibrosis), y según el genotipo del VHC. Desde luego hay mas variables que manejar en la consulta clínica como puede ser la valoración clínica de las manifestaciones extrahepáticas, grado de replicación del virus, necesidad de trasplante, embarazo y un largo etc.…. En este trabajo de tesis uno de los aspectos en los que hemos incidido ha sido la importancia de la correcta determinación del genotipo del VHC.Tesis Univ. Granada.Tesis. Departamento de Medicin

A Multi-Omics Approach Reveals New Signatures in Obese Allergic Asthmatic Children

Asthma is a multifactorial condition where patients with identical clinical diagnoses do not have the same clinical history or respond to treatment. This clinical heterogeneity is reflected in the definition of two main endotypes. We aimed to explore the metabolic and microbiota signatures that characterize the clinical allergic asthma phenotype in obese children. Methods: We used a multi-omics approach combining clinical data, plasma and fecal inflammatory biomarkers, metagenomics, and metabolomics data in a cohort of allergic asthmatic children. Results: We observed that the obese allergic asthmatic phenotype was markedly associated with higher levels of leptin and lower relative proportions of plasma acetate and a member from the Clostridiales order. Moreover, allergic children with a worse asthma outcome showed higher levels of large unstained cells, fecal D lactate and D/L lactate ratio, and with a higher relative proportion of plasma creatinine and an unclassified family member from the RF39 order belonging to the Mollicutes class. Otherwise, children with persistent asthma presented lower levels of plasma citrate and dimethylsulfone. Conclusion: Our integrative approach shows the molecular heterogeneity of the allergic asthma phenotype while highlighting the use of omics technologies to examine the clinical phenotype at a more holistic level.Fundación Progreso y Salud Project PI-0373-2014Redes temáticas de investigación cooperativa RETIC Red SAMID RD12/0026/001

Modulation of faecal metagenome in Crohn's disease: Role of microRNAs as biomarkers.

The gut microbiota plays a key role in the maintenance of intestinal homeostasis and the development and activation of the host immune system. It has been shown that commensal bacterial species can regulate the expression of host genes. 16S rRNA gene sequencing has shown that the microbiota in inflammatory bowel disease (IBD) is abnormal and characterized by reduced diversity. MicroRNAs (miRNAs) have been explored as biomarkers and therapeutic targets, since they are able to regulate specific genes associated with Crohn's disease (CD). In this work, we aim to investigate the composition of gut microbiota of active treatment-naïve adult CD patients, with miRNA profile from gut microbiota. To investigate the composition of gut microbiota of active treatment-naïve adult CD patients, with miRNA profile from gut microbiota. Patients attending the outpatient clinics at Valme University Hospital without relevant co-morbidities were matched according to age and gender. Faecal samples of new-onset CD patients, free of treatment, and healthy controls were collected. Faecal samples were homogenized, and DNA was amplified by PCR using primers directed to the 16S bacterial rRNA gene. Pyrosequencing was performed using GS-Junior platform. For sequence analysis, MG-RAST server with the database Ribosomal Project was used. MiRNA profile and their relative abundance were analyzed by quantitative PCR. Microbial community was characterized using 16S rRNA gene sequencing in 29 samples (n = 13 CD patients, and n = 16 healthy controls). The mean Shannon diversity was higher in the healthy control population compared to CD group (5.5 vs 3.7). A reduction in Firmicutes and an increase in Bacteroidetes were found. Clostridia class was also significantly reduced in CD. Principal components analysis showed a grouping pattern, identified in most of the subjects in both groups, showing a marked difference between control and CD groups. A functional metabolic study showed that a lower metabolism of carbohydrates (P = 0.000) was found in CD group, while the metabolism of lipids was increased. In CD patients, three miRNAs were induced in affected mucosa: mir-144 (6.2 ± 1.3 fold), mir-519 (21.8 ± 3.1) and mir-211 (2.3 ± 0.4). Changes in microbial function in active non-treated CD subjects and three miRNAs in affected vs non-affected mucosa have been found. miRNAs profile may serve as a biomarker