Rafael Borrás

High prevalence of subtype 4 among isolates of Blastocystis hominis from symptomatic patients of a health district of Valencia (Spain)

In order to know the genetic diversity of Blastocystis hominis from a health district of Valencia (Spain) 51 clinical isolates from symptomatic patients,31 axenic and 20 monoxenic, were ribotyped by analysing the restriction fragment length polymorphism (RFLP) of amplicons obtained by polymerase chain reaction (PCR) of small-subunit of ribosomal DNA genes (SSU-rDNA). For this purpose, DNA was subjected to two independent PCR (RD3-RD5, F1-R1) and to three independent treatments with restrictases (AluI, HinfI and RsaI). The digested DNA was separated electrophoretically, the isolates were clustered into ribotypes (ribodemes, RD3-RD5; subgroups, F1-R1) according to their profiles and the results were translated into genetic subtypes (ST) proposed by a consensus terminology. The results show that the isolates studied are an heterogeneous population and that both PCR-RFLP SSU-rDNA protocols have a similar discriminative power, since it allowed the ribotyping of all isolates and their clustering into four demes: ribodemes 1, 3 and 3-r and 6, which include isolates belonging to subgroup III, IV, V and V-r, respectively; which were assigned to ST1 (2%), ST2 (3.9%) and ST4 (94.1%). The most common of which is a zoonotic subtype (Blastocystis ratti) which includes, according to recent studies, non-pathogenic and pathogenic variants.

Diagnostic accuracy and potential clinical value of the LightCycler SeptiFast assay in the management of bloodstream infections occurring in neutropenic and critically ill patients

OBJECTIVES The objectives of this study were to compare the performance of the LightCycler SeptiFast Test MGRADE and conventional blood culture in the etiological diagnosis of febrile episodes occurring in neutropenic and critically ill patients (in the intensive care unit; ICU), and to assess the potential clinical value of the SeptiFast test in patient management. METHODS A total of 86 febrile episodes occurring in 33 neutropenic patients and 53 ICU patients were analyzed. Blood samples for blood culture and SeptiFast testing were obtained at the onset of fever, before the implementation of empirical antimicrobial therapy. RESULTS The overall microorganism-to-isolate agreement between the SeptiFast test and blood culture was 69% (κ=0.37) in neutropenic patients and 75% (κ=0.56) in ICU patients. The sensitivity of the SeptiFast assay for clinically relevant episodes of bacteremia and fungemia was 62% in neutropenic patients and 70% in ICU patients. Based on SeptiFast results, empirical treatments were deemed adequate in all but one of the febrile episodes. Nevertheless, early antibiotic treatment readjustment was judged feasible in most of clinically significant episodes overall. CONCLUSIONS The SeptiFast assay is a valuable ancillary method for the diagnosis of bloodstream infections in neutropenic and ICU patients. In these clinical settings, results of the SeptiFast assay may lead to a more targeted antibiotic therapy early after the onset of fever.

Search of Chemical Scaffolds for Novel Antituberculosis Agents

A method to identify chemical scaffolds potentially active against Mycobacterium tuberculosis is presented. The molecular features of a set of structurally heterogeneous antituberculosis drugs were coded by means of structural invariants. Three techniques were used to obtain equations able to model the antituberculosis activity: linear discriminant analysis, multilinear regression, and shrinkage estimation–ridge regression. The model obtained was statistically validated through leave-n-out test, and an external set and was applied to a database for the search of new active agents. The selected compounds were assayed in vitro, and among those identified as active stand reserpine, N,N,N′,N′-tetrakis-(2-pyridylmethyl)-ethylenediamine (TPEN), trifluoperazine, pentamidine, and 2-methyl-4,6-dinitro-phenol (DNOC). They show activity comparable to or superior to ethambutol, used in combination with other drugs for the prevention and treatment of Mycobacterium avium complex and drug-resistant tuberculosis.

Diagnósticos al alta hospitalaria de las personas inmigrantes en la ciudad de Valencia (2001-2002)

Fundamento: La inmigracion en Espana se ha incrementado de forma notable en la ultima decada. Por razones teoricas se ha planteado que esta poblacion pudiera ser responsable de un aumento de la incidencia y/o transmision de enfermedades infecciosas. Sin embargo, existen escasos estudios acerca de las enfermedades de este colectivo que permitan conocer la importancia de esta afirmacion. El objetivo de este estudio es cuantificar la atencion hospitalaria a esta poblacion en la ciudad de Valencia. Metodos: Se obtuvieron los registros de altas de los Servicios de Admision de los hospitales publicos de la ciudad de Valencia. Se desarrollo un estudio observacional retrospectivo para el ano comprendido entre el 1 de octubre de 2001 al 30 de septiembre del 2002. Resultados: Recuperamos 8.444 ingresos que identificaron 1.577 ingresos de inmigrantes. Se evidencio una proporcion similar de inmigrantes registrados con documentacion reglada (841, 51,16%) e inmigrantes supuestamente no regularizados (803, 48,84%). Predominaron las mujeres (68,3%) frente a los varones (31,7%) y una media de edad inferior a los 30 anos. El diagnostico de alta mas frecuente fue «complicaciones del embarazo, parto y puerperio» con 37,7% del total. Los «traumatismos y envenenamientos», «enfermedades del aparato digestivo» y «enfermedades del aparato respiratorio» representaron el 12,9%, el 7,8% y el 5,4%, respectivamente. Las «enfermedades infecciosas y parasitarias» supusieron el 4,4%. Conclusiones: La proporcion de enfermedades infecciosas e infecto-contagiosas observadas no contribuyen a alentar las opiniones difundidas sobre importacion o reemergencia de enfermedades desde los colectivos de inmigrantes. La inmigracion masiva acaecida requiere multiples adaptaciones del Sistema de Salud que permitan establecer el perfil de salud de este colectivo, basado en estimaciones poblacionales.

Occurrence and genotypes of Giardia isolated from lambs in Spain

Three hundred and eighty six faecal specimens were randomly collected from 1- to 3-month-old lambs from 16 farms in Spain to investigate the presence of different genotypes of Giardia duodenalis. Individual specimens were examined by IFA (Immunofluorescence assay) and beta-giardin PCR polymerase chain reaction. Cysts of G. duodenalis were shed by lambs in every flock analyzed, showing a prevalence by farms of 100%. The average prevalence of G. duodenalis for the 386 specimens was 42%, ranging from 8.3 to 80% depending on the farm. beta-giardin PCR positive samples were sequenced to determine the genotypes present at each farm and seven new subtypes of beta-giardin Assemblage E are reported in this study. In each farm, one to six different beta-giardin subtypes were found, showing the high variability of the target. Also, one flock had the zoonotic Assemblage A. This is the first report of Giardia subgenotype A-1 in sheep in Spain.

Positive Result of the Aspergillus Galactomannan Antigen Assay Using Bronchoalveolar Lavage Fluid from a Patient with an Invasive Infection Due to Lichtheimia ramosa

Positive galactomannan antigen (GM) test results with serum by the Platelia Aspergillus assay (Bio-Rad, Marnes-La-Coquette, France) have been reported to occur during invasive fungal infections caused by Penicillium marneffei ([8][1]), Cryptococcus neoformans ([2][2]), Geotrichum capitatum ([3][3

Performance of the LightCycler SeptiFast Test Mgrade in Detecting Microbial Pathogens in Purulent Fluids

ABSTRACT The performance of the LightCycler SeptiFast (SF) assay was compared to that of culture methods in the detection of microorganisms in 43 purulent fluids from patients with pyogenic infections. The SF assay was more sensitive than the culture methods (86% versus 61%, respectively), irrespective of whether the infections were mono- or polymicrobial.

Performance characteristics of the new Abbott Real Time MTB assay for detection of Mycobacterium tuberculosis complex in respiratory specimens

The performance of the Abbott Real Time MTB assay for detection of Mycobacterium tuberculosis complex in respiratory specimens was evaluated using a standard culture as the reference. The overall concordance between both methods was 0.95. The assay displayed an excellent sensitivity (100% for smear-positive/92.3% for smear-negative specimens) and specificity (100%).

Frequent detection of cytomegalovirus (CMV) DNA in the lower respiratory tract in CMV‐seropositive pediatric patients with underlying chronic bronchopulmonary diseases lacking canonical immunosuppression

Cytomegalovirus (CMV) may be a relevant cause of morbidity in patients displaying various inflammatory diseases. In this study, it was investigated whether CMV DNA is detected in the lower respiratory tract and the systemic compartment in pediatric patients with chronic or recurrent bronchopulmonary diseases. A total of 42 lower respiratory tract specimens and 11 paired plasma samples from 42 patients were analyzed for the presence of CMV DNA by real‐time PCR. The respiratory specimens were also screened for the presence of respiratory viruses and human herpesvirus 6 (HHV‐6) and 7 (HHV‐7) by PCR methods. Quantitative bacterial and fungal cultures were performed. IL‐6 levels in the respiratory specimens were quantified using ELISA. CMV DNA was detected either in the lower respiratory airways, in plasma, or both in 54.5% of CMV‐seropositive patients. The levels of IL‐6 were significantly higher in these patients than in those with no detectable levels of CMV DNA. HHV‐6 and HHV‐7 DNA were detected in three and one patients, respectively. Respiratory viruses were detected in 13 of the 42 patients. Significant growth of one or more bacterial species was observed in 17 patients. No significant association was found between the presence of CMV DNA and the detection of other microorganisms. The data indicated that the presence of CMV DNA in the lower respiratory tract is a frequent finding in children with chronic or recurrent bronchopulmonary diseases. Further, prospective observational studies are needed to assess the impact of this phenomenon, if any, on the clinical course of these patients. J. Med. Virol. 85:888–892, 2013.

Chemotherapy of Aspergillus fumigatus keratitis: An experimental study

AbstractAn experimental Keratitis study of Aspergillus fumigatus was performed in 130 rabbits divided into 12 groups of ten animals each. Three antifungal drugs (myconazole, amphotericin B and pimaricin) were tested using two procedures (topical drops and subconjunctival injections) and two different concentrations (500 and 10 000 times the MIC). In each case, the drugs were applied every 3 h starting 14 h after inoculation. Miconazole was useful at 10 mg/ ml concentration by topical drops and subconjunctival injections, but was less useful at 5 mg/ ml. Amphotericin B was useful at 5 mg/ ml concentration by topical drops and less useful at 2 mg/ ml.No differences were found between the two concentrations by subconjunctival administration. Pimaricin was useful by topical drops at 50 mg/ ml concentration and less useful at 10 mg/ ml as well as by subconjunctival injections.