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We screened 526 wild small mammals for zoonotic viruses in northwest Spain and found hantavirus in common voles (Microtus arvalis) (1.5%) and high prevalence (48%) of orthopoxvirus among western Mediterranean mice (Mus spretus). We also detected arenavirus among small mammals. These findings suggest novel risks for viral transmission in the region.
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Doenças Transmissíveis , Infecções por Hantavirus , Orthohantavírus , Vírus de RNA , Doenças dos Roedores , Animais , Arvicolinae , Orthohantavírus/genética , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/veterinária , Mamíferos , Camundongos , Doenças dos Roedores/epidemiologia , Espanha/epidemiologia , Zoonoses Virais , Zoonoses/epidemiologiaRESUMO
The natural host ranges of many viruses are restricted to very specific taxa. Little is known about the molecular barriers between species that lead to the establishment of this restriction or generally prevent virus emergence in new hosts. Here, we identify genomic polymorphisms in a natural rodent host associated with a strong genetic barrier to the transmission of European Tula orthohantavirus (TULV). We analysed the very abrupt spatial transition between two major phylogenetic clades in TULV across the comparatively much wider natural hybrid zone between evolutionary lineages of their reservoir host, the common vole (Microtus arvalis). Genomic scans of 79,225 single nucleotide polymorphisms (SNPs) in 323 TULV-infected host individuals detected 30 SNPs that were consistently associated with the TULV clades CEN.S or EST.S in two replicate sampling transects. Focusing the analysis on 199 voles with evidence of genomic admixture at the individual level (0.1-0.9) supported statistical significance for all 30 loci. Host genomic variation at these SNPs explained up to 37.6% of clade-specific TULV infections. Genes in the vicinity of associated SNPs include SAHH, ITCH and two members of the Syngr gene family, which are involved in functions related to immune response or membrane transport. This study demonstrates the relevance of natural hybrid zones as systems not only for studying processes of evolutionary divergence and speciation, but also for the detection of evolving genetic barriers for specialized parasites.
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Infecções por Hantavirus , Orthohantavírus , Vírus de RNA , Animais , Arvicolinae/genética , FilogeniaRESUMO
Global warming is predicted to have major effects on the annual time windows during which species may successfully reproduce. At the organismal level, climatic shifts engage with the control mechanism for reproductive seasonality. In mammals, laboratory studies on neuroendocrine mechanism emphasize photoperiod as a predictive cue, but this is based on a restricted group of species. In contrast, field-oriented comparative analyses demonstrate that proximate bioenergetic effects on the reproductive axis are a major determinant of seasonal reproductive timing. The interaction between proximate energetic and predictive photoperiodic cues is neglected. Here, we focused on photoperiodic modulation of postnatal reproductive development in common voles (Microtus arvalis), a herbivorous species in which a plastic timing of breeding is well documented. We demonstrate that temperature-dependent modulation of photoperiodic responses manifest in the thyrotrophin-sensitive tanycytes of the mediobasal hypothalamus. Here, the photoperiod-dependent expression of type 2 deiodinase expression, associated with the summer phenotype was enhanced by 21°C, whereas the photoperiod-dependent expression of type 3 deiodinase expression, associated with the winter phenotype, was enhanced by 10°C in spring voles. Increased levels of testosterone were found at 21°C, whereas somatic and gonadal growth were oppositely affected by temperature. The magnitude of these temperature effects was similar in voles photoperiodical programmed for accelerated maturation (ie, born early in the breeding season) and in voles photoperiodical programmed for delayed maturation (ie, born late in the breeding season). The melatonin-sensitive pars tuberalis was relatively insensitive to temperature. These data define a mechanistic hierarchy for the integration of predictive temporal cues and proximate thermo-energetic effects in mammalian reproduction.
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Arvicolinae/fisiologia , Gônadas/fisiologia , Fotoperíodo , Reprodução , Estações do Ano , Temperatura , Animais , Ritmo Circadiano , Metabolismo Energético , Feminino , Iodeto Peroxidase , Masculino , MelatoninaRESUMO
OBJECTIVES: Potential effect of three environmental variables (population density of rodents, global weather in the form of the North Atlantic Oscillation (NAO) index, and acorn crop) on human morbidity rate of three zoonoses: tick-borne encephalitis (TBE), leptospirosis and tularaemia were analysed in the Czech Republic for the period 1970-1990. METHODS: The Pearson's correlation analysis was used. RESULTS: The correlation analysis revealed that the significant factor for explaining annual morbidity rates of these zoonoses was the abundance of common voles (Microtus arvalis) in the current year (for leptospirosis) or in the previous calendar year (for TBE and tularaemia). CONCLUSIONS: The two other environmental variables tested (NAO index and acorn production) do not seem to play a significant role in these zoonoses in Central Europe.
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Encefalite Transmitida por Carrapatos , Leptospirose , Tularemia , Animais , Encefalite Transmitida por Carrapatos/epidemiologia , Incidência , Leptospirose/epidemiologia , Tempo (Meteorologia)RESUMO
Using long-term data on incidences of Lyme disease and tickborne encephalitis, we showed that the dynamics of both diseases in central Europe are predictable from rodent host densities and climate indices. Our approach offers a simple and effective tool to predict a tickborne disease risk 1 year in advance.
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Clima , Ratos/fisiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Animais , Reservatórios de Doenças , Encefalite Transmitida por Carrapatos/epidemiologia , Encefalite Transmitida por Carrapatos/etiologia , Europa Oriental/epidemiologia , Incidência , Ixodes , Doença de Lyme/epidemiologia , Doença de Lyme/etiologia , Fatores de Risco , Doenças Transmitidas por Carrapatos/etiologiaRESUMO
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.
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Arvicolinae/microbiologia , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/microbiologia , Animais , Bartonella , Infestações por Pulgas , Francisella tularensis , Humanos , Prevalência , Espanha/epidemiologiaRESUMO
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.
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Arvicolinae , Bactérias/isolamento & purificação , Coinfecção/veterinária , Doenças dos Roedores/epidemiologia , Zoonoses/epidemiologia , Animais , Coinfecção/epidemiologia , Coinfecção/microbiologia , Vetores de Doenças , Feminino , Masculino , Densidade Demográfica , Prevalência , Doenças dos Roedores/microbiologia , Espanha/epidemiologia , Zoonoses/microbiologiaRESUMO
Compounds and products in the biocide and plant protection sector can only be registered after formal risk assessment to ensure safety for users and the environment. In bird and mammal risk assessment, this is routinely done using generic focal species as models, which are of particular exposure risk. Such a species is the common vole (Microtus arvalis) due to its high food intake relative to the low body weight. For wild species, biological samples, data and hence realistic exposure estimations are particularly difficult to obtain. In recent years, advances have been made in the techniques related to serial microsampling of laboratory mice and rats that allow for a reduction in sampling volumes. Similar progress in wild species sampling is missing. This study presents a proof of concept to dose wild rodents with relevant compounds and to draw serial, low volume blood samples suitable for state-of-the art toxicokinetic analyses. For the first time, the jugular vein of common voles was used to administer compounds (two frequently used fungicidal components). This procedure and the following microsampling of blood (2 × 10 µl six times within 24 hours) from the lateral tail vein did not affect body weight and mortality of voles. Samples were sufficient to detect dissipation patterns of the compounds from blood in toxicokinetic analysis. These results suggest that microsampling can be well translated from laboratory mice to wild rodent species and help to obtain realistic exposure estimates in wild rodents for ecotoxicological studies as well as to promote the 3R concept in studies with wild rodent species.
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Arvicolinae/sangue , Coleta de Amostras Sanguíneas/métodos , Dioxóis/toxicidade , Ecotoxicologia/métodos , Exposição Ambiental/efeitos adversos , Monitoramento Ambiental/métodos , Fungicidas Industriais/toxicidade , Pirimidinas/toxicidade , Pirróis/toxicidade , Administração Intravenosa , Animais , Dioxóis/administração & dosagem , Dioxóis/sangue , Dioxóis/farmacocinética , Feminino , Fungicidas Industriais/administração & dosagem , Fungicidas Industriais/sangue , Fungicidas Industriais/farmacocinética , Masculino , Pirimidinas/administração & dosagem , Pirimidinas/sangue , Pirimidinas/farmacocinética , Pirróis/administração & dosagem , Pirróis/sangue , Pirróis/farmacocinética , Reprodutibilidade dos Testes , Medição de Risco , ToxicocinéticaRESUMO
Strong spatiotemporal variation in population size often leads to reduced genetic diversity limiting the adaptive potential of individual populations. Key genes of adaptive variation are encoded by the immune genes of the major histocompatibility complex (MHC) playing an essential role in parasite resistance. How MHC variation persists in rodent populations that regularly experience population bottlenecks remains an important topic in evolutionary genetics. We analysed the consequences of strong population fluctuations on MHC class II DRB exon 2 diversity in two distant common vole (Microtus arvalis) populations in three consecutive years using a high-throughput sequencing approach. In 143 individuals, we detected 25 nucleotide alleles translating into 14 unique amino acid MHC alleles belonging to at least three loci. Thus, the overall allelic diversity and amino acid distance among the remaining MHC alleles, used as a surrogate for the range of pathogenic antigens that can be presented to T-cells, are still remarkably high. Both study populations did not show significant population differentiation between years, but significant differences were found between sites. We concluded that selection processes seem to be strong enough to maintain moderate levels of MHC diversity in our study populations outcompeting genetic drift, as the same MHC alleles were conserved between years. Differences in allele frequencies between populations might be the outcome of different local parasite pressures and/or genetic drift. Further understanding of how pathogens vary across space and time will be crucial to further elucidate the mechanisms maintaining MHC diversity in cyclic populations.
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Arvicolinae/genética , Deriva Genética , Variação Genética/genética , Genética Populacional , Complexo Principal de Histocompatibilidade/genética , Seleção Genética/genética , Animais , Frequência do Gene , FilogeniaRESUMO
Phylogenetic reconstruction of the evolutionary history of closely related organisms may be difficult because of the presence of unsorted lineages and of a relatively high proportion of heterozygous sites that are usually not handled well by phylogenetic programs. Genomic data may provide enough fixed polymorphisms to resolve phylogenetic trees, but the diploid nature of sequence data remains analytically challenging. Here, we performed a phylogenomic reconstruction of the evolutionary history of the common vole (Microtus arvalis) with a focus on the influence of heterozygosity on the estimation of intraspecific divergence times. We used genome-wide sequence information from 15 voles distributed across the European range. We provide a novel approach to integrate heterozygous information in existing phylogenetic programs by repeated random haplotype sampling from sequences with multiple unphased heterozygous sites. We evaluated the impact of the use of full, partial, or no heterozygous information for tree reconstructions on divergence time estimates. All results consistently showed four deep and strongly supported evolutionary lineages in the vole data. These lineages undergoing divergence processes split only at the end or after the last glacial maximum based on calibration with radiocarbon-dated paleontological material. However, the incorporation of information from heterozygous sites had a significant impact on absolute and relative branch length estimations. Ignoring heterozygous information led to an overestimation of divergence times between the evolutionary lineages of M. arvalis. We conclude that the exclusion of heterozygous sites from evolutionary analyses may cause biased and misleading divergence time estimates in closely related taxa.
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Arvicolinae/genética , Especiação Genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Animais , Teorema de Bayes , Evolução Molecular , Haplótipos , Heterozigoto , Modelos Genéticos , Filogenia , Polimorfismo de Nucleotídeo ÚnicoRESUMO
In highly fluctuating populations with complex social systems, genetic patterns are likely to vary in space and time due to demographic and behavioural processes. Cyclic rodents are extreme examples of demographically instable populations that often exhibit strong social organization. In such populations, kin structure and spacing behaviour may vary with density fluctuations and impact both the composition and spatial structure of genetic diversity. In this study, we analysed the multiannual genetic structure of a cyclic rodent, Microtus arvalis, using a sample of 875 individuals trapped over three complete cycles (from 1999 to 2007) and genotyped at 10 microsatellite loci. We tested the predictions that genetic diversity and gene flow intensity vary with density fluctuations. We found evidences for both spatial scale-dependant variations in genetic diversity and higher gene flow during high density. Moreover, investigation of sex-specific relatedness patterns revealed that, although dispersal is biased toward males in this species, distances moved by both sexes were lengthened during high density. Altogether, these results suggest that an increase in migration with density allows to restore the local loss of genetic diversity occurring during low density. We then postulate that this change in migration results from local competition, which enhances female colonization of empty spaces and male dispersal among colonies.
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Arvicolinae/genética , Fluxo Gênico , Variação Genética , Genética Populacional , Distribuição Animal , Animais , Feminino , França , Genótipo , Masculino , Repetições de Microssatélites , Densidade Demográfica , Dinâmica PopulacionalRESUMO
BACKGROUND: The common vole is an agricultural pest and population outbreaks have caused significant crop damage in the agricultural areas of the west-central part of the Iberian Peninsula (Spain) during the last few decades. Thus, monitoring is imperative to gain a comprehensive insight on its spatiotemporal habitat colonization dynamics. This work was performed on a long-term database with the primary objective of establishing an operational framework for understanding the spatial and temporal distribution of the common vole, all of it based on the Getis-Ord statistics. RESULTS: The temporal evolution of the vole abundance index (VAI) was consistent for the three studied habitats, that is, crops, reservoirs and paths. Furthermore, the majority of common vole abundance peaks coincide with summer periods, especially in annual herbaceous crops. The spatial distribution of vole abundance exhibited a non-random pattern, characterized by spatial clustering. Particularly, the areas with higher significance of this clustering were located at the so-called 'Tierra de Campos' county, covering northern Valladolid, southern Palencia and north-eastern Zamora provinces. Periods of major incidence and concentration were 2013-2014, 2016-2017 and 2019. Common vole temporal occupancy patterns demonstrate that colonization occurred simultaneously in alfalfa reservoirs and dispersion paths, both preceding settlement in annual herbaceous crop plots. CONCLUSION: The geographic information system (GIS)-hotspots-based methodology proposed here can be valuable for stakeholders involved in integrated management of the common vole, serving as a detector of pest-prone areas in both space and time. These hotspots are useful for predicting future surveillance areas that accurately reflect pest colonization patterns. It was found that common vole abundance along dispersal paths acts as a source of dissemination, preceding the risk of colonization in annual herbaceous crop plots. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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BACKGROUND: Worldwide, pest rodents can cause extensive damage to agriculture, forestry, food storage, and infrastructure and pose a risk to public health and livestock due to the spread of zoonotic pathogens. In Europe, the most common pest rodent species is the common vole (Microtus arvalis). Management during periodic outbreaks largely relies on rodenticidal bait with zinc phosphide. Efficient baiting with rodenticides or possibly anti-fertility products in the future require baiting methods that allow a sufficient proportion of the population to consume an effective dose of bait. We used a bait with the quantitative marker ethyl-iophenoxic acid (Et-IPA) to evaluate baiting strategies in enclosure experiments. This wheat-based bait with Et-IPA was placed in bait boxes or directly into the tunnel system entrances in different seasons and common vole abundances. Voles were live-trapped, individually marked and blood samples were collected to relate Et-IPA blood residues to bait uptake. RESULTS: The percentage of animals consuming bait was not heavily affected by the baiting strategy but voles had higher Et-IPA blood residues if tunnel baiting was used in autumn and if bait boxes were used in winter. Non-reproductive as well as lighter animals tended to have higher Et-IPA blood residues than reproductive individuals, whereas sex had no effect. Population density had a negative effect on the probability of residues present as well as on Et-IPA blood concentration. CONCLUSION: The results of this study might help to improve baiting techniques to manage overabundant rodent pest species regardless of the compounds to be delivered. © 2024 Julius Kühn-Institut. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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Despite the large number of studies on rodent ectoparasites-most of them vectors of epidemiologically important pathogens-infestation patterns remain poorly understood in various ecological contexts, such as the highly patchy agricultural landscapes. We aimed to relate the infestation of rodents to temporal, habitat, and host variables. We assessed the difference in parasite prevalence and mean abundance depending on host sex, age, and body weight, season, and land use intensity. Furthermore, we analysed the effect of host species abundance and the differential responses of parasites in main and minor host species. The field survey was conducted in a rural landscape in southern Transylvania (Romania) between June and September 2010-2011. We live-trapped small mammals, collected the ticks and fleas, and recorded the presence of lice and mites. Overall, we found the same infestation patterns largely reported in the literature: higher prevalence and mean abundance in heavier adult males, significant seasonality and differences among host species, and evidence of the dilution effect. The uniqueness of our study system was the negative effect of the land use intensity on the prevalence and mean abundance of parasites, explained by the highly patchy mosaic landscape.
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Evolutionary divergence of viruses is most commonly driven by co-divergence with their hosts or through isolation of transmission after host shifts. It remains mostly unknown, however, whether divergent phylogenetic clades within named virus species represent functionally equivalent byproducts of high evolutionary rates or rather incipient virus species. Here, we test these alternatives with genomic data from two widespread phylogenetic clades in Tula orthohantavirus (TULV) within a single evolutionary lineage of their natural rodent host, the common vole Microtus arvalis. We examined voles from forty-two locations in the contact region between clades for TULV infection by reverse transcription (RT)-PCR. Sequencing yielded twenty-three TULV Central North and twenty-one TULV Central South genomes, which differed by 14.9-18.5 per cent at the nucleotide and 2.2-3.7 per cent at the amino acid (AA) level without evidence of recombination or reassortment between clades. Geographic cline analyses demonstrated an abrupt (<1 km wide) transition between the parapatric TULV clades in continuous landscape. This transition was located within the Central mitochondrial lineage of M. arvalis, and genomic single nucleotide polymorphisms showed gradual mixing of host populations across it. Genomic differentiation of hosts was much weaker across the TULV Central North to South transition than across the nearby hybrid zone between two evolutionary lineages in the host. We suggest that these parapatric TULV clades represent functionally distinct, incipient species, which are likely differently affected by genetic polymorphisms in the host. This highlights the potential of natural viral contact zones as systems for investigating the genetic and evolutionary factors enabling or restricting the transmission of RNA viruses.
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Cyclic rodent populations exhibit pronounced changes in body mass associated with the population cycle phase, long-known as Chitty effect. Although Chitty effect is a common epiphenomenon in both America and Europe, there is still incomplete evidence about the generality of these patterns across the entire range of most species. Moreover, despite decades of research, the underlying factors driving Chitty effect remains poorly understood. Here, we examined the influence of intrinsic and extrinsic factors that may underlie observed patterns in vole size variation in the Iberian common vole Microtus arvalis asturianus. We weighed and measured 2816 adult voles that were captured during 6 trapping periods. Vole numbers and body mass showed strong period- and phase-related variation both in females and males, demonstrating marked Chitty effect in the studied population. Body mass of adult males correlated with body length, evidencing that heavier males are also structurally larger. Statistical models showed that probability of occurrence of large-sized vole (> 37 g) was significantly more likely in reproductive males, during increase and peak phases, and it was modulated by habitat, with crop fields and field margins between crops showing an increased likelihood. We suggest an effect of the habitat on vole body mass mediated by predation.
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Arvicolinae , Peso Corporal , Ecossistema , Animais , Arvicolinae/fisiologia , Masculino , Feminino , Tamanho Corporal/fisiologia , Dinâmica PopulacionalRESUMO
Oceanic islands have been a test ground for evolutionary theory, but here, we focus on the possibilities for evolutionary study created by offshore islands. These can be colonized through various means and by a wide range of species, including those with low dispersal capabilities. We use morphology, modern and ancient sequences of cytochrome b (cytb) and microsatellite genotypes to examine colonization history and evolutionary change associated with occupation of the Orkney archipelago by the common vole (Microtus arvalis), a species found in continental Europe but not in Britain. Among possible colonization scenarios, our results are most consistent with human introduction at least 5100 bp (confirmed by radiocarbon dating). We used approximate Bayesian computation of population history to infer the coast of Belgium as the possible source and estimated the evolutionary timescale using a Bayesian coalescent approach. We showed substantial morphological divergence of the island populations, including a size increase presumably driven by selection and reduced microsatellite variation likely reflecting founder events and genetic drift. More surprisingly, our results suggest that a recent and widespread cytb replacement event in the continental source area purged cytb variation there, whereas the ancestral diversity is largely retained in the colonized islands as a genetic 'ark'. The replacement event in the continental M. arvalis was probably triggered by anthropogenic causes (land-use change). Our studies illustrate that small offshore islands can act as field laboratories for studying various evolutionary processes over relatively short timescales, informing about the mainland source area as well as the island.
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Arvicolinae/genética , Evolução Biológica , Genética Populacional , Ilhas , Distribuição Animal , Animais , Arvicolinae/anatomia & histologia , Teorema de Bayes , Bélgica , Citocromos b/genética , DNA Mitocondrial/genética , Repetições de Microssatélites , Dados de Sequência Molecular , Filogenia , Reino UnidoRESUMO
Background: Over 200 different animal species are susceptible to infection by tularemia, highly infectious disease caused by Francisella tularensis. Hares (Lepus sp.) and small rodents, such as common vole (Microtus arvalis), have been acknowledged as the most significant sources of human tularemia infection in most European countries. Our objective was to verify ability of these species in predicting incidence of human tularemia in a Central European country, the Czech Republic. Materials and Methods: We used 17 years of data on densities of European hare (Lepus europaeus) and common vole, and climate variability to test effects of these factors on temporal dynamics of tularemia incidence. The data were obtained from annual reports available from online e-repositories. Results: The analysis showed that 33% of the yearly variation in human tularemia incidence was explained solely by the abundance of European hare in the Czech Republic during 2007-2022. Density of common vole and North Atlantic Oscillation index, a measure of climate variability, did not significantly explain tularemia incidence. While hare population declined severely during 1993-2022, we did not detect any clear accompanied decrease in the prevalence of tularemia in humans and hares. Conclusion: Contrary to expectations, only hares proved capable in predicting yearly dynamics in human tularemia incidence in the Czech Republic. We call for continued monitoring of infection rates in hares and advocate the use of hunter estimates of hare abundance as a cheap and effective means of predicting the risk of tularemia.
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Francisella tularensis , Lebres , Tularemia , Animais , Humanos , Tularemia/epidemiologia , Tularemia/veterinária , República Tcheca/epidemiologia , Densidade Demográfica , Roedores , ArvicolinaeRESUMO
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.
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Between June and August 2014, 45 cases of leptospirosis were notified among workers on two strawberry farms in North-West Germany. We describe the characteristics of the outbreak and the actions taken to prevent further cases. The activities of the local, federal and national public health and veterinary authorities included collection of case data, laboratory testing of human specimens and of small mammals trapped on the fields, investigation of weather data, as well as information provided to farmers, field workers, physicians and to the authorities in Poland and Romania. Of the 45 identified cases (median age 22, 60% male), 47% were hospitalized. Characteristic symptoms were fever ≥38.5°C, generalized muscle pain and an increase in renal or liver enzymes. Thirteen cases were laboratory confirmed by serological and/or molecular methods. ELISA tests for Leptospira IgG and IgM-antibodies were positive in those samples taken >5 days after hospitalization. The probable causative agent was identified as Leptospira kirschneri serovar Grippotyphosa. Leptospira-specific DNA was found in kidneys of 67% of 64 trapped small mammals and was further identified as Leptospira kirschneri multi locus sequence type 110. During the estimated time period of human infections, the affected region faced warm weather with heavy rainfalls. The results of this investigation are in accordance with the theory of a chain of infection from mice to field workers during warm and humid weather. In 2015, a campaign was initiated to inform physicians, farmers and workers to enhance prevention measures, such as the use of personal protective equipment and early consultation of physicians in case of illness. Since then, no further outbreak occurred.