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[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.

More than 1000 humans have acquired the febrile disease tularemia in Spain since the first notification of human cases in 1997. We here aimed to study the recent molecular evolution of the causative bacterium Francisella tularensis during disease establishment in Spain. Single-nucleotide polymorphisms (SNPs) and variable-number tandem repeats (VNTRs) were analyzed in whole-genome sequences (WGS) of F. tularensis. Short-read WGS data for 20 F. tularensis strains from humans infected in the periods 2014-2015 and 2018-2020 in Spain were generated. These data were combined with WGS data of 25 Spanish strains from 1998 to 2008 and two reference strains. Capillary electrophoresis data of VNTR genetic regions were generated and compared with the WGS data for the 11 strains from 2014 to 2015. Evolutionary relationships among strains were analyzed by phylogenetic methods. We identified 117 informative SNPs in a 1,577,289-nucleotide WGS alignment of 47 F. tularensis genomes. Forty-five strains from Spain formed a star-like SNP phylogeny with six branches emerging from a basal common node. The most recently evolved genomes formed four additional star-like structures that were derived from four branches of the basal common node. VNTR copy number variation was detected in two out of 10 VNTR regions examined. Genetic clustering of strains by VNTRs agreed with the clustering by SNPs. The SNP data provided higher resolution among strains than the VNTRs data in all but one cases. There was an excellent correlation between VNTR marker sizing by capillary electrophoresis and prediction from WGS data. The genetic data strongly support that tularemia, indeed, emerged recently in Spain. Distinct genetic patterns of local F. tularensis population expansions imply that the pathogen has colonized a previously disease-free geographical area. We also found that genome-wide SNPs provide higher genetic resolution among F. tularensis genomes than the use of VNTRs, and that VNTR copy numbers can

Objetivo: La presente revisión de la literatura tiene como objetivo principal conocer la efectividad de los Recubrimientos pulpares directos e indirectos en dientes primarios, además de conocer las diferentes técnicas de tratamiento, los factores que influyen en la eficacia del mismo y los criterios pronósticos. Material y métodos: Se realizó una búsqueda bibliográfica en las bases de datos Pubmed, FAMA y Cochrane Libary. Aplicando los criterios de inclusión y exclusión, se seleccionaron 10 artículos finalmente para llevar a cabo los resultados y la discusión. Conclusión: Los recubrimientos pulpares en dientes temporales se consideran una técnica correcta, efectiva y reproducible en la clínica. Se necesitan más ensayos y estudios clínicos a largo plazo que demuestren la eficacia de unos medicamentos pulpares frente a otros además de otros factores incluidos en el estudio., Objective: The main objective of this literature review is to know the effectiveness of direct and indirect pulp coatings on primary teeth, in addition to knowing the different treatment techniques, the factors that influence its efficacy and prognostic criteria. Material and methods: A bibliographic search was performed in the Pubmed, FAMA and Cochrane Libary databases. Applying the inclusion and exclusion criteria, 10 articles were finally selected to carry out the results and discussion. Conclusion: Pulp coverings in temporary teeth are considered a correct, effective and reproducible technique in the clinic. More trials and long-term clinical studies are needed to demonstrate the efficacy of some pulp medications against others in addition to other factors included in the study.

Diseases and host dynamics are linked, but their associations may vary in strength, be time-lagged, and depend on environmental influences. Where a vector is involved in disease transmission, its dynamics are an additional influence, and we often lack a general understanding on how diseases, hosts and vectors interact. We report on the occurrence of six zoonotic arthropod-borne pathogens (Anaplasma, Bartonella, Borrelia, Coxiella, Francisella and Rickettsia) in common voles (Microtus arvalis) throughout a population fluctuation and how their prevalence varies according to host density, seasonality and vector prevalence. We detected Francisella tularensis and four species of Bartonella, but not Anaplasma, Borrelia, Coxiella or Rickettsia. Bartonella taylorii and B. grahamii prevalence increased and decreased with current host (vole and mice) density, respectively, and increased with flea prevalence. Bartonella doshiae prevalence decreased with mice density. These three Bartonella species were also more prevalent during winter. Bartonella rochalimae prevalence varied with current and previous vole density (delayed-density dependence), but not with season. Coinfection with F. tularensis and Bartonella occurred as expected from the respective prevalence of each disease in voles. Our results highlight that simultaneously considering pathogen, vector and host dynamics provide a better understanding of the epidemiological dynamics of zoonoses in farmland rodents.

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.

Tularemia in humans in northwestern Spain is associated with increases in vole populations. Prevalence of infection with Francisellatularensis in common voles increased to 33% during a vole population fluctuation. This finding confirms that voles are spillover agents for zoonotic outbreaks. Ecologic interactions associated with tularemia prevention should be considered.

Pearls: Host population dynamics are the key of wildlife zoonotic risk; Aquatic and terrestrial agents of tularemia coexist in nature; Irrigation has provided aquatic reservoirs and a grass-loving amplification agent for tularemia in Spain; Fluctuating mammalian populations shape tularemia epidemiology; Tularemia surveillance must target unstable mammalian host populations.

For many infections transmitting to humans from reservoirs in nature, disease dispersal patterns over space and time are largely unknown. Here, a reversed genomics approach helped us understand disease dispersal and yielded insight into evolution and biological properties of Francisella tularensis, the bacterium causing tularemia. We whole-genome sequenced 67 strains and characterized by single-nucleotide polymorphism assays 138 strains, collected from individuals infected 1947-2012 across Western Europe. We used the data for phylogenetic, population genetic and geographical network analyses. All strains (n= 205) belonged to a monophyletic population of recent ancestry not found outside Western Europe. Most strains (n= 195) throughout the study area were assigned to a star-like phylogenetic pattern indicating that colonization of Western Europe occurred via clonal expansion. In the East of the study area, strains were more diverse, consistent with a founder population spreading from east to west. The relationship of genetic and geographic distance within the F. tularensis population was complex and indicated multiple long-distance dispersal events. Mutation rate estimates based on year of isolation indicated null rates; in outbreak hotspots only, there was a rate of 0.4 mutations/genome/year. Patterns of nucleotide substitution showed marked AT mutational bias suggestive of genetic drift. These results demonstrate that tularemia has moved from east to west in Europe and that F. tularensis has a biology characterized by long-range geographical dispersal events and mostly slow, but variable, replication rates. The results indicate that mutation-driven evolution, a resting survival phase, genetic drift and long-distance geographical dispersal events have interacted to generate genetic diversity within this species.

En el presente trabajo se lleva a cabo un análisis de la situación actual del sector vitivinícola en Francia. Francia se encuentra entre los líderes en este sector a nivel mundial, siendo líder en cuanto a nivel de producción, en superficie destinada al viñedo, en consumo y en volumen y valor de sus exportaciones. En primer lugar, se establecen el conjunto de factores que influyen en el sector del vino en Francia. Estos factores son determinantes en nuestro análisis puesto que se trata de las características que definen la situación actual del sector. Entre estos factores destaca la producción, la estructura empresarial, el consumo, el precio o los canales de distribución utilizados en el mercado del vino en Francia. En segundo lugar, se ha realizado un análisis del comercio exterior en Francia, estableciendo primero la situación de los indicadores económicos fundamentales en el análisis del comercio internacional y posteriormente analizando las exportaciones e importaciones desarrolladas en Francia en el año 2015. Dentro de las mismas hemos establecido una división que diferencia entre categoría de producto y país de destino o procedencia. Dentro de dicha división, de nuevo establecemos una distinción entre volumen y valor de las exportaciones e importaciones. Posteriormente, como síntesis del estudio realizado, se han detallado una serie de perspectivas del mercado del vino en Francia. Para ello se ha desarrollado un escueto análisis de la coyuntura internacional para establecer más correctamente la situación del mercado vitivinícola francés a nivel mundial. Finalmente, se extraen una serie de conclusiones fruto del análisis desarrollado