Influence of immigration on tuberculosis transmission patterns in Castellón, Spain (2004–2007)

Objetivo Describir los patrones de transmisión de la tuberculosis en Castellón en un periodo de importantes cambios demográficos. Métodos Estudio prospectivo descriptivo de los pacientes con cultivo positivo en la provincia de Castellón entre 2004 y2007. Se describen los pacientes por año y nacionalidad, y se cotejan con los casos declarados a la Dirección General de Salud Pública (DGSP). Se estudia la población con patrón molecular disponible por RFLP (restriction fragment length polymorphism) y se analizan las variables de los pacientes agrupados (clusters) a partir de los datos de la DGSP y del Programa de Gestión del Laboratorio. Resultados Según la DGSP, la tasa global de tuberculosis por 100.000 habitantes en la provincia de Castellón fue de 15,7 en 2004, 19,9 en 2005, 18,2 en 2006 y 17,5 en 2007. En nuestro laboratorio se identificaron las cepas de 301 pacientes, que suponen el 77% (301/390) de los casos declarados y el 94% (301/321) aquellos con cultivo positivo. El porcentaje de tuberculosis en extranjeros aumentó hasta superar el 50% en 2007. Se disponía de estudio molecular en el 95% de los casos (286), con un 58% de españoles y un 42% de extranjeros. El porcentaje de agregación fue del 40%, con un 30% de clusters mixtos. Según el estudio convencional de contactos, el 85% de los pacientes en cluster habían sido considerados casos aislados. Conclusiones El aumento de la tasa de tuberculosis en Castellón se debe, principalmente, al creciente número de los casos en extranjeros. Disponer del estudio molecular de todos los pacientes con cultivo positivo nos ha permitido analizar cómo y dónde se transmite la tuberculosis. El 40% de los pacientes se agruparon en clusters, y eran mixtos un tercio de ellos, lo que indica una elevada integración de los inmigrantes.Background This study aimed to identify tuberculosis transmission patterns in Castellón in a period of major demographic changes. Methods A prospective study of patients with positive culture in the province of Castellon over a 4-year period (2004–2007) was carried out. Cases were described by year and nationality and were compared with those reported to the Department of Public Health. We studied the population with available molecular patterns, identified through restriction fragment length polymorphism (RFLP) and analyzed the variables from patient clusters, based on data collected in surveys of the Department of Health and the Laboratory Management Program. Results According to data from the Department of Public Health, the overall rate of tuberculosis per 100,000 inhabitants in the province of Castellón was 15.7 in 2004, 19.9 in 2005, 18.2 in 2006 and 17.5 in 2007. In our laboratory, strains were identified from 301 patients, representing 77% (301/390) of reported cases and 94% (301/321) of reported cases with a positive culture. The percentage of tuberculosis among foreigners increased with age, exceeding 50% in 2007. Molecular studies were available in 95% of patients (286); 58% were Spanish and 42% were foreigners, of whom 54% were Romanians. The cluster percentage was 40%, with 30% of mixed clusters. According to conventional contact studies, 85% of patients in clusters had been considered isolated cases. Conclusions The increased rate of tuberculosis in Castellón was mainly due to the increasing number of cases among foreigners, mostly Romanians. The availability of molecular studies in all patients with a positive culture allowed us to analyze how and where tuberculosis is transmitted in our province. Forty percent of the patients were grouped into clusters; of these, mixed clusters accounted for one third, indicating the high integration of immigrants in our area

Absceso hepático invasivo recidivante por Klebsiella mucoviscidosa

Liver abscess is an infection with polymicrobial etiology, but the emergence of Klebsiella pneumoniae as the main cause, specifically serotypes K1 or K2 with a mucoid phenotype, is increasing in Europe. We present here the case of a 76 years old man who consulted due to fever. He had a history of two previous admissions for liver abscesses secondary to Klebsiella pneumoniae. The abdominopelvic CT showed a new liver abscess and the blood cultures were positive, again, for K. pneumoniae, confirming mucoviscid serotype K1.El absceso hepático es una infección con etiología polimicrobiana, pero la emergencia de Klebsiella pneumoniae como principal causante, y en concreto los serotipos K1 o K2 con fenotipo mucoide, está aumentando en Europa. Se presenta el caso de un varón de 76 años, que consultó por cuadro febril. Presentaba antecedentes de dos ingresos previos por abscesos hepáticos secundarios a Klebsiella pneumoniae. El TAC abdomino-pélvico evidenció un nuevo absceso hepático y los hemocultivos fueron positivos, nuevamente, para K. pneumoniae, constatándose serotipo mucoviscidoso K1

Wastewater monitoring of a community COVID-19 outbreak in a Spanish municipality

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) wastewater surveillance has become an increasingly important tool since the initial moments of the coronavirus disease (COVID-19) pandemic, mainly conducted at a large population scale. However, the large-sized sewersheds may not provide suitable information for monitoring localized outbreaks. After the declaration of a community COVID-19 outbreak in two neighborhoods of Castellón (Spain), SARS-CoV-2 RNA levels were monitored in wastewater samples. Moreover, genomic sequencing was performed. Thirty-three samples were collected in 2020, distributed over three points of the sewage network, two of which were close to the areas where the cases were declared. The third point was located at the inlet of the municipal wastewater treatment plant (WWTP). The samples were analyzed by RT-qPCR, using specific N1 and N2 target regions. The sum of confirmed cases, with the date of symptoms onset within the 3 weeks before each sampling day, was calculated. SARS-CoV-2 RNA was detected on most days in the two sampling points at neighborhood level, and the detection became negative when the number of cases with symptoms onset during the last 21 days in the study areas decreased to 0 or 1 case. The genomic sequencing performed for RNA from wastewater and clinical samples showed the same variant. The detection of SARS-CoV-2 and the subsequent non-detection provided the epidemiologists in charge of controlling the outbreak with useful information to confirm its closure, complementing the clinical and epidemiological data. Our findings illustrate the value of wastewater surveillance for localized outbreaks, especially in situations of low incidence of COVID-19 at the broader community level

Role of the first WHO mutation catalogue in the diagnosis of antibiotic resistance in Mycobacterium tuberculosis in the Valencia Region, Spain: a retrospective genomic analysis

9 páginas, 2 figuras, 1 tablaBackground: In June, 2021, WHO published the most complete catalogue to date of resistance-conferring mutations in Mycobacterium tuberculosis. Here, we aimed to assess the performance of genome-based antimicrobial resistance prediction using the catalogue and its potential for improving diagnostics in a real low-burden setting. Methods: In this retrospective population-based genomic study M tuberculosis isolates were collected from 25 clinical laboratories in the low-burden setting of the Valencia Region, Spain. Culture-positive tuberculosis cases reported by regional public health authorities between Jan 1, 2014, and Dec 31, 2016, were included. The drug resistance profiles of these isolates were predicted by the genomic identification, via whole-genome sequencing (WGS), of the high-confidence resistance-causing variants included in the catalogue and compared with the phenotype. We determined the minimum inhibitory concentration (MIC) of the isolates with discordant resistance profiles using the resazurin microtitre assay. Findings: WGS was performed on 785 M tuberculosis complex culture-positive isolates, and the WGS resistance prediction sensitivities were: 85·4% (95% CI 70·8–94·4) for isoniazid, 73·3% (44·9–92·2) for rifampicin, 50·0% (21·1–78·9) for ethambutol, and 57·1% (34·0–78·2) for pyrazinamide; all specificities were more than 99·6%. Sensitivity values were lower than previously reported, but the overall pan-susceptibility accuracy was 96·4%. Genotypic analysis revealed that four phenotypically susceptible isolates carried mutations (rpoB Leu430Pro and rpoB Ile491Phe for rifampicin and fabG1 Leu203Leu for isoniazid) known to give borderline resistance in standard phenotypic tests. Additionally, we identified three putative resistance-associated mutations (inhA Ser94Ala, katG Leu48Pro, and katG Gly273Arg for isoniazid) in samples with substantially higher MICs than those of susceptible isolates. Combining both genomic and phenotypic data, in accordance with the WHO diagnostic guidelines, we could detect two new multidrug-resistant cases. Additionally, we detected 11 (1·6%) of 706 isolates to be monoresistant to fluoroquinolone, which had been previously undetected. Interpretation: We showed that the WHO catalogue enables the detection of resistant cases missed in phenotypic testing in a low-burden region, thus allowing for better patient-tailored treatment. We also identified mutations not included in the catalogue, relevant at the local level. Evidence from this study, together with future updates of the catalogue, will probably lead in the future to the partial replacement of culture testing with WGS-based drug susceptibility testing in our setting. Funding: European Research Council and the Spanish Ministerio de Ciencia.This project received funding from the European Research Council under the European Union’s Horizon 2020 Research and Innovation Program Grant 101001038 (TB-RECONNECT; awarded to IC), from Ministerio de Ciencia (Spanish Government) Project PID2019-104477RB-I00 (awarded to IC), and from Generalitat Valenciana Project AICO/2018/113 (awarded to IC). AMG-M is funded by a Formación deProfesorado Universitario grant programme (FPU19/04562) from Ministerio de Universidades (Spanish Government). IC is also supported by the European Commission–NextGenerationEU, through Centro Superior de Investigaciones Científicas Global Health Platform (PTI Salud Global). We thank all the members of the Valencia RegionTuberculosis Working Group

Modelización de las condiciones de transporte y conservación prolongada en frutas y hortalizas

Este documento se generó a partir de la VI Reunión de la Red Temática FRUTURA de CYTED realizada en la ciudad de Buenos Aires, Argentina del 26 al 30 de Setiembre de 2011. Organizado por el Laboratorio de Calidad y Postcosecha de Frutas y Hortalizas de la E.E.A. Balcarce del INTA, el programa de esta reunión se enmarcó dentro del principal objetivo de la Red, que es el desarrollo de un sistema integral de mejora de la calidad y seguridad de las frutas durante la manipulación, el transporte y la comercialización, mediante nuevas tecnologías de inspección y monitorización

Population-based sequencing of Mycobacterium tuberculosis reveals how current population dynamics are shaped by past epidemics

23 páginas, 4 figuras, 1 tabla.Transmission is a driver of tuberculosis (TB) epidemics in high-burden regions, with assumed negligible impact in low-burden areas. However, we still lack a full characterization of transmission dynamics in settings with similar and different burdens. Genomic epidemiology can greatly help to quantify transmission, but the lack of whole genome sequencing population-based studies has hampered its application. Here, we generate a population-based dataset from Valencia region and compare it with available datasets from different TB-burden settings to reveal transmission dynamics heterogeneity and its public health implications. We sequenced the whole genome of 785 Mycobacterium tuberculosis strains and linked genomes to patient epidemiological data. We use a pairwise distance clustering approach and phylodynamic methods to characterize transmission events over the last 150 years, in different TB-burden regions. Our results underscore significant differences in transmission between low-burden TB settings, i.e., clustering in Valencia region is higher (47.4%) than in Oxfordshire (27%), and similar to a high-burden area as Malawi (49.8%). By modeling times of the transmission links, we observed that settings with high transmission rate are associated with decades of uninterrupted transmission, irrespective of burden. Together, our results reveal that burden and transmission are not necessarily linked due to the role of past epidemics in the ongoing TB incidence, and highlight the need for in-depth characterization of transmission dynamics and specifically tailored TB control strategies.European Research Council 638553-TB-ACCELERATE; European Research Council 101001038-TBRECONNECT; Ministerio de Ciencia e Innovación SAF2016-77346-RPeer reviewe

Evolución de la Tuberculosis en Castellón (2008-2012). Caracterización genotípica mediante 15 MIRU-VNTR

La tuberculosis (TB) es considerada una de las primeras enfermedades infecciosas humanas de las que se tiene constancia y aún hoy, en pleno siglo XXI, es una de las más importantes causantes de morbilidad y mortalidad en todo el mundo. Según describe la Organización Mundial de la Salud (OMS) en su 19º informe, 9 millones de personas enfermaron de TB a nivel mundial en el año 2013, aunque en éste se refiere que ha habido una disminución de casos en los últimos años. También el Centro Europeo para la Prevención y Control de Enfermedades (ECDC) y el Centro Nacional de Epidemiología (CNE) informan que en Europa y en España, en el año 2012, el número de casos de TB disminuyó respecto al año anterior. La presente Tesis Doctoral tiene como objetivos conocer la evolución de la TB en la provincia de Castellón a lo largo del periodo comprendido entre enero de 2008 a diciembre de 2012, describir las características clínico-epidemiológicas y microbiológicas de los casos diagnosticados con cultivo positivo, detallar la frecuencia de resistencias de Mycobacterium tuberculosis frente a fármacos tuberculostáticos de primera línea, y analizar los patrones de transmisión de TB en Castellón durante los años 2010, 2011 y 2012 mediante tipificación molecular, utilizando la técnica 15 MIRU-VNTR. Para conseguir los 3 primeros objetivos consultamos varias fuentes de información como son el Sistema de Información del Laboratorio (SIL), el Sistema del Análisis de la Vigilancia Epidemiológica (AVE) y los informes de TB editados por la Generalitat Valenciana. Para llevar a cabo el cuarto objetivo, analizamos las cepas congeladas y recuperadas de los tres años indicados mediante la técnica de epidemiología molecular 15 MIRU-VNTR. En la provincia de Castellón se declararon 387 casos de TB, de los que 320 (82,7%) tuvieron el cultivo positivo; en 2008 se diagnosticaron microbiológicamente 74 casos, 83 en el 2009, 66 en el 2010, 56 en el 2011 y 41 en el 2012. En cuanto a las características de los pacientes, la razón hombre/mujer fue de 1,9 y la edad media fue de 32,5 años, concentrándose el 60,6% en el grupo de adultos jóvenes (15-44 años). El 43,7% de los pacientes eran extranjeros, el 93,8% fueron casos no tratados previamente, el 29,4% presentaba al menos un factor de riesgo para desarrollar enfermedad tuberculosa y, por último, el 67,2% de los pacientes fueron hospitalizados. Respecto a las formas clínicas, un 87,2% fueron TB pulmonar, predominando la ganglionar entre las formas extrapulmonares. A partir del estudio convencional de contactos se sospechó 7 posibles brotes de 2 miembros. El estudio de sensibilidad se realizó a las 320 cepas aisladas, resultando resistentes el 13,5%, siendo la resistencia a isoniazida del 8,1%. En población de origen español el porcentaje de resistencias fue del 10,6% mientras que en extranjeros fue del 15,7%. Se realizó la tipificación molecular a 151 cepas de las 163 aisladas en los tres años. Los loci más polimórficos fueron el VNTR 4052, el VNTR 2163b y el VNTR 3690, y los que presentaron un poder de discriminación elevado, calculado a partir del índice de diversidad de Hunter-Gaston, fueron los MIRU 26, 40 y 10, y los VNTR 577, 2401, 3690, 2163b, 1955 y 4052. Evidenciamos que 122 cepas presentaron un patrón único y que 29 cepas se agruparon en 12 clusters, siendo el porcentaje de agrupación del 19,2% y la tasa de transmisión reciente de un 11,3%. En el presente estudio se concluye que la tasa de TB, y paralelamente el número de casos con cultivo positivo, en la provincia de Castellón ha disminuido a lo largo de los años analizados; que el fenómeno de la inmigración tiene una influencia importante en la TB de nuestra provincia; que el número de hospitalizaciones, aún siendo menor que en estudios previos, sigue siendo elevado y que la resistencia global de un 13,5% es también más elevada a la descrita en trabajos anteriores en Castellón. De estas resistencias, la de isoniazida superior al 5% obliga a tratar la TB en los dos primeros meses con asociación de 4 fármacos, como recomienda la OMS; el porcentaje elevado de resistencia a isoniazida en el año 2010 fue debido a un brote que fue confirmado con tipificación molecular. Aplicando la técnica 15 MIRU-VNTR, con un índice de discriminación elevado de 0,998, se ha detectado una baja tasa de transmisión durante los 3 años analizados

Imported taeniasis in the context of colon hydrotherapy

Key Clinical Message We report a case of imported taeniasis, under the modality of “visiting friends and relatives”, in the context of colon hydrotherapy. This technique allows the detection of proglottids, diagnosed in this case as Taenia saginata based on the gravid proglottid that presented more than 13 uterine branches and showed active motility. Moreover, the patient did not consume pork for religious reasons. The treatment with paromomicin sulfate was effective. In this case, a trip to Ethiopia, together with the ingestion of raw beef, was the cause of parasitization. It is highly advisable to obtain detailed information from the patient on their background, especially their travel and dietary histories

Is there a widespread clone of Serratia marcescens producing outbreaks worldwide?

[Background]: Serratia marcescens frequently causes outbreaks in healthcare settings. There are few studies using high-throughput sequencing (HTS) that analyse S. marcescens outbreaks. We present the analysis of two outbreaks in neonatal intensive care units (NICUs) in hospitals from the Comunitat Valenciana (CV, Spain) and the impact of using different reference genomes.[Methods]: DNA from cultured isolates was extracted and sequenced by HTS using Illumina NextSeq. Reads were mapped against two reference genomes, strains UMH9 and Db11, and the unmapped fraction of the genomes was assembled to fully genetically characterize the isolates.[Findings]: Isolates from the first outbreak were identical to the UMH9 reference, an unrelated isolate obtained three years earlier in the USA. This did not occur when the Db11 strain, a standard reference for S. marcescens, was used as the reference for mapping. To check whether UMH9 was a widely distributed clone spreading in the CV, the second outbreak isolates were mapped against this reference. They were not closely related to this strain, and this outbreak could be defined as such regardless of the reference used for mapping the reads.[Conclusions]: The choice of the reference for genomic analysis of outbreaks is a critical decision. In the case of the first outbreak, this choice changed the interpretation of the results drastically, allowing or preventing the definition of the outbreak according to the reference used. Although HTS is a powerful tool for epidemiological analysis, it is still essential to collect microbiological and epidemiological data for the correct interpretation of the results.This work was supported by the Spanish Ministry of Science, Innovation, and Universities (BFU2014-58656-R and BFU2017-89594-R), and by Generalitat Valenciana (Prometeo2016/122). CFC was supported by the Spanish Ministry of Science, Innovation, and Universities (BES-2015-074204). The infrastructures and instrumental for the FISABIO Sequencing Service were funded by the European Regional Development Fund (ERDF).Peer reviewe