Strengthening the genomic surveillance of Francisella tularensis by using culture-free whole-genome sequencing from biological samples

[Introduction] Francisella tularensis is a highly infectious bacterium that causes the zoonotic disease tularemia. The development of genotyping methods, especially those based on whole-genome sequencing (WGS), has recently increased the knowledge on the epidemiology of this disease. However, due to the difficulties associated with the growth and isolation of this fastidious pathogen in culture, the availability of strains and subsequently WGS data is still limited.[Methods] To surpass these constraints, we aimed to implement a culture-free approach to capture and sequence F. tularensis genomes directly from complex samples. Biological samples obtained from 50 common voles and 13 Iberian hares collected in Spain were confirmed as positive for F. tularensis subsp. holarctica and subjected to a WGS target capture and enrichment protocol, using RNA oligonucleotide baits designed to cover F. tularensis genomic diversity.[Results] We obtained full genome sequences of F. tularensis from 13 animals (20.6%), two of which had mixed infections with distinct genotypes, and achieved a higher success rate when compared with culture-dependent WGS (only successful for two animals). The new genomes belonged to different clades commonly identified in Europe (B.49, B.51 and B.262) and subclades. Despite being phylogenetically closely related to other genomes from Spain, the detected clusters were often found in other countries. A comprehensive phylogenetic analysis, integrating 599 F. tularensis subsp. holarctica genomes, showed that most (sub)clades are found in both humans and animals and that closely related strains are found in different, and often geographically distant, countries.[Discussion] Overall, we show that the implemented culture-free WGS methodology yields timely, complete and high-quality genomic data of F. tularensis, being a highly valuable approach to promote and potentiate the genomic surveillance of F. tularensis and ultimately increase the knowledge on the genomics, ecology and epidemiology of this highly infectious pathogen.This work was supported by funding from the European Union EU4Health Programme under grant agreement No 101113460 (GENEO) and by the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT). The acquisition of WGS-related reagents was supported by the Ministerio de Economía y Competitividad, Government of Spain/FEDER, RESERTULA project (CLG2015-66962-C2-2-R). Fieldwork for sample collection was funded by the Ministerio de Economía y Competitividad, Government of Spain/FEDER ECOTULA project (CGL2015-66962-C2-1-R) and by the Ministerio de Ciencia, Innovación y Universidades BOOMRAT project (PID2019-109327RB-I00) from Spain.Peer reviewe

4to. Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad. Memoria académica

Este volumen acoge la memoria académica de la Cuarta edición del Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad, CITIS 2017, desarrollado entre el 29 de noviembre y el 1 de diciembre de 2017 y organizado por la Universidad Politécnica Salesiana (UPS) en su sede de Guayaquil. El Congreso ofreció un espacio para la presentación, difusión e intercambio de importantes investigaciones nacionales e internacionales ante la comunidad universitaria que se dio cita en el encuentro. El uso de herramientas tecnológicas para la gestión de los trabajos de investigación como la plataforma Open Conference Systems y la web de presentación del Congreso http://citis.blog.ups.edu.ec/, hicieron de CITIS 2017 un verdadero referente entre los congresos que se desarrollaron en el país. La preocupación de nuestra Universidad, de presentar espacios que ayuden a generar nuevos y mejores cambios en la dimensión humana y social de nuestro entorno, hace que se persiga en cada edición del evento la presentación de trabajos con calidad creciente en cuanto a su producción científica. Quienes estuvimos al frente de la organización, dejamos plasmado en estas memorias académicas el intenso y prolífico trabajo de los días de realización del Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad al alcance de todos y todas

Análisis bibliométrico de la investigación realizada en el Departamento de Historia Contemporánea

Sin resume

Zoonotic bacteria in fleas parasitizing common voles, northwestern Spain

Research Letter.We detected Francisella tularensis and Bartonella spp. in fleas parasitizing common voles (Microtus arvalis) from northwestern Spain; mean prevalence was 6.1% for F. tularensis and 51% for Bartonella spp. Contrasted vector–host associations in the prevalence of these bacteria suggest that fleas have distinct roles in the transmission cycle of each pathogen in nature.This work was supported by ECOVOLE (Factores ecologicos que influyen en la reproducción y dinámica poblacional del topillo campesino (Microtus arvalis) en medios agrarios; CGL2012-35348), ECOTULA (Ecología de la Tularemia: dinámica espacio-temporal, ciclos ecológicos de transmisión y mapas de riesgo en ecosistemas agrarios del NO de España; CGL2015-66962-C2-1-R), and RESERTULA (Microbiología de la Tularemia: circulación de Francisella tularensis en los ecosistemas agrarios del NO de España. Estudio de relaciones epidemiológicas y filogenéticas; CLG2015-66962-C2-2-R) projects funded by the Government of Spain (lMINECO/FEDER). R.R.-P. was supported by a PhD studentship from the University of Valladolid (co-funded by Banco Santander, RR 30/04/2014).Peer reviewe

Strengthening the genomic surveillance of Francisella tularensis by using culture-free whole-genome sequencing from biological samples

Introduction: Francisella tularensis is a highly infectious bacterium that causes the zoonotic disease tularemia. The development of genotyping methods, especially those based on whole-genome sequencing (WGS), has recently increased the knowledge on the epidemiology of this disease. However, due to the difficulties associated with the growth and isolation of this fastidious pathogen in culture, the availability of strains and subsequently WGS data is still limited. Methods: To surpass these constraints, we aimed to implement a culture-free approach to capture and sequence F. tularensis genomes directly from complex samples. Biological samples obtained from 50 common voles and 13 Iberian hares collected in Spain were confirmed as positive for F. tularensis subsp. holarctica and subjected to a WGS target capture and enrichment protocol, using RNA oligonucleotide baits designed to cover F. tularensis genomic diversity. Results: We obtained full genome sequences of F. tularensis from 13 animals (20.6%), two of which had mixed infections with distinct genotypes, and achieved a higher success rate when compared with culture-dependent WGS (only successful for two animals). The new genomes belonged to different clades commonly identified in Europe (B.49, B.51 and B.262) and subclades. Despite being phylogenetically closely related to other genomes from Spain, the detected clusters were often found in other countries. A comprehensive phylogenetic analysis, integrating 599 F. tularensis subsp. holarctica genomes, showed that most (sub)clades are found in both humans and animals and that closely related strains are found in different, and often geographically distant, countries. Discussion: Overall, we show that the implemented culture-free WGS methodology yields timely, complete and high-quality genomic data of F. tularensis, being a highly valuable approach to promote and potentiate the genomic surveillance of F. tularensis and ultimately increase the knowledge on the genomics, ecology and epidemiology of this highly infectious pathogen.The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by funding from the European Union EU4Health Programme under grant agreement No 101113460 (GENEO) and by the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT). The acquisition of WGS-related reagents was supported by the Ministerio de Economía y Competitividad, Government of Spain/FEDER, RESERTULA project (CLG2015-66962-C2-2-R). Fieldwork for sample collection was funded by the Ministerio de Economía y Competitividad, Government of Spain/FEDER ECOTULA project (CGL2015-66962-C2-1-R) and by the Ministerio de Ciencia, Innovación y Universidades BOOMRAT project (PID2019-109327RB-I00) from Spain.S

Molecular evidence of Francisella tularensis in nature, 10 years after a large outbreak of tularemia in Spain

Resumen del trabajo presentado a la 9th International Conference on Tularemia, celebrada en Montréal (Canada) del 16 al 19 de octubre de 2018.[Background and aim]: Tularemia is a zoonosis caused by Francisella tularensis, a pathogen that can use a variety of transmission routes in nature as well as in humans. Since 1997 more than 1,000 human cases of tularemia have been declared in Spain in two major outbreaks, but knowledge about the epidemiology of this infection is very limited. The ability of F. tularensis to find mechanisms to persist in the environment and cause new outbreaks makes it necessary to identify, among others, the potential reservoirs during inter-epizootic periods, the main objective of this study. [Material and methods]: The sampling points were located in an area of Castilla y León, well known as “hot spots” for tularemia. They corresponded to surface water and sediments, as well as crayfish, mosquitoes, hares and ticks. Specifically, 8 zones divided into 4 subzones were sampled, including three rivers, three irrigation channels and three lagunes. Samples were collected in July and November of 2016 and 2017. For detection of Francisella lpnA was amplified by conventional PCR and hybridization by reverse line blotting with specific probes. Positive samples were confirmed by a real-time multitarget TaqMan PCR, using tul4 and ISFtu2 assays. For additional characterization sdhA gen and VNTR Ft-M19 were amplified and sequenced. [Results]: A total of 102 samples of water, 111 sediments, 343 crayfish, 156 mosquitos, 64 hares and 126 ticks from 14 hares were collected. Overall, 2 samples of water, 3 of sediments, 7 hares and 17 ticks tested positive for Francisella. Interestingly, F. tularensis subspecie holarctica was detected in water, hares and ticks while F. hispaniensis was the species detected in water and sediments. [Conclusions]: In this study we provide evidence of Francisella presence in nature during periods in which no large outbreaks of tularemia occur, and suggest that hares and ticks play an important role in the maintenance of the bacteria in the terrestrial environment. Moreover, to the best of our knowledge this is the first description of F. hispaniensis in abiotic environment, which enforces the need to assess the risk for the population and establish the necessary measures for its prevention and control.Peer Reviewe

Terapia de estrógenos vaginales y riesgo de recurrencia en mujeres con antecedente de cáncer de mama: Vaginal estrogen therapy and the risk of recurrence in women with a history of breast cancer

Breast cancer remains the most common malignant neoplasm and one of the leading causes of mortality in women, making it a significant target for global health efforts and a public health priority. Through the use of innovative therapies, survival rates have improved, leading to the emergence of associated conditions such as genitourinary menopausal syndrome. Hormonal therapy is employed for managing this condition, significantly alleviating its symptoms and, in some cases, serving as the sole solution. The most commonly utilized approach is vaginal estrogen therapy. Nevertheless, there have been reports of a potential risk of breast cancer recurrence associated with its use. In the Spanish-speaking context, there is limited evidence discussing this topic. A search was conducted across PubMed, ScienceDirect, and MEDLINE databases, using the terms "Vaginal Estrogen Therapy," "Recurrence," and "Breast Cancer." It was determined that, on a global scale, vaginal estrogen therapy is an effective and safe therapeutic option for managing genitourinary menopausal syndrome in women with a history of breast cancer. This therapy does not appear to increase the risk of recurrence, with the exception of those undergoing treatment with aromatase inhibitors. For these individuals, alternative therapies are recommended to mitigate this potential risk.El cáncer de mama sigue siendo la neoplasia maligna más frecuente y mortal en mujeres, considerándose un importante objetivo de la salud global y prioridad en salud pública. Con el uso de terapias innovadoras, ha mejorado la supervivencia, apareciendo condiciones asociadas, como el síndrome genitourinario menopaúsico. La terapia hormonal, se utiliza para el manejo de esta condición, mejorando sustancialmente la sintomatología, e incluso, siendo en algunos casos la única solución. La más utilizada, es la terapia de estrógenos vaginales. Sin embargo, se ha descrito un posible riesgo de recurrencia de cáncer de mama con su uso. En habla hispana, no existe evidencia que haya discutido este tópico. Se llevó a cabo una búsqueda en las bases PubMed, ScienceDirect y MEDLINE, utilizando los términos “Terapia de estrógenos vaginales”, “Recurrencia” y “Cáncer de mama”. Se encontró, que, de forma global, la terapia de estrógenos vaginales es una opción terapéutica eficaz y segura en el manejo del síndrome genitourinario menopaúsico en mujeres con antecedente de cáncer de mama, sin incrementar el riesgo de recurrencia, a excepción de aquellas tratadas con inhibidores de la aromatasa, en quienes se recomienda el uso de otras terapias para evitar acarrear este riesgo

La historia de Europa en el aula virtual: materiales multimedia y elaboración de vídeos didácticos para la enseñanza de la historia de Europa

Se pretende desarrollar un nuevo modelo de docencia de la historia de Europa de los siglos XIX y XX mediante la elaboración de materiales multimedia y edición de vídeos docentes que permitan complementar el desarrollo de las clases presenciales

Table_1_Strengthening the genomic surveillance of Francisella tularensis by using culture-free whole-genome sequencing from biological samples.XLSX

IntroductionFrancisella tularensis is a highly infectious bacterium that causes the zoonotic disease tularemia. The development of genotyping methods, especially those based on whole-genome sequencing (WGS), has recently increased the knowledge on the epidemiology of this disease. However, due to the difficulties associated with the growth and isolation of this fastidious pathogen in culture, the availability of strains and subsequently WGS data is still limited.MethodsTo surpass these constraints, we aimed to implement a culture-free approach to capture and sequence F. tularensis genomes directly from complex samples. Biological samples obtained from 50 common voles and 13 Iberian hares collected in Spain were confirmed as positive for F. tularensis subsp. holarctica and subjected to a WGS target capture and enrichment protocol, using RNA oligonucleotide baits designed to cover F. tularensis genomic diversity.ResultsWe obtained full genome sequences of F. tularensis from 13 animals (20.6%), two of which had mixed infections with distinct genotypes, and achieved a higher success rate when compared with culture-dependent WGS (only successful for two animals). The new genomes belonged to different clades commonly identified in Europe (B.49, B.51 and B.262) and subclades. Despite being phylogenetically closely related to other genomes from Spain, the detected clusters were often found in other countries. A comprehensive phylogenetic analysis, integrating 599 F. tularensis subsp. holarctica genomes, showed that most (sub)clades are found in both humans and animals and that closely related strains are found in different, and often geographically distant, countries.DiscussionOverall, we show that the implemented culture-free WGS methodology yields timely, complete and high-quality genomic data of F. tularensis, being a highly valuable approach to promote and potentiate the genomic surveillance of F. tularensis and ultimately increase the knowledge on the genomics, ecology and epidemiology of this highly infectious pathogen.</p