An Update of Evidence for Pathogen Transmission by Ticks of the Genus Hyalomma.

Current and likely future changes in the geographic distribution of ticks belonging to the genus Hyalomma are of concern, as these ticks are believed to be vectors of many pathogens responsible for human and animal diseases. However, we have observed that for many pathogens there are no vector competence experiments, and that the level of evidence provided by the scientific literature is often not sufficient to validate the transmission of a specific pathogen by a specific Hyalomma species. We therefore carried out a bibliographical study to collate the validation evidence for the transmission of parasitic, viral, or bacterial pathogens by Hyalomma spp. ticks. Our results show that there are very few validated cases of pathogen transmission by Hyalomma tick species.

Design theory to better target public health priorities: An application to Lyme disease in France.

In the context of complex public health challenges led by interdependent changes such as climate change, biodiversity loss, and resistance to treatment, it is important to mobilize methods that guide us to generate innovative interventions in a context of uncertainty and unknown. Here, we mobilized the concept-knowledge (CK) design theory to identify innovative, cross-sectoral, and cross-disciplinary research and design programs that address the challenges posed by tick-borne Lyme disease in France, which is of growing importance in the French public health and healthcare systems. Within the CK methodological framework, we developed an iterative approach based on literature analysis, expert interviews, analysis of active French research projects, and work with CK experts to contribute to design "an action plan against Lyme disease." We produced a CK diagram that highlights innovative concepts that could be addressed in research projects. The outcome is discussed within four areas: (i) effectiveness; (ii) environmental sustainability in prevention actions; (iii) the promotion of constructive involvement of citizens in Lyme challenges; and (iv) the development of care protocols for chronic conditions with an unknown diagnosis. Altogether, our analysis questioned the health targets ranging from population to ecosystem, the citizen involvement, and the patient consideration. This means integrating social and ecological science, as well as the multidisciplinary medical patient journey, from the start. CK theory is a promising framework to assist public health professionals in designing programs for complex yet urgent contexts, where research and data collection are still not sufficient to provide clear guidance.

The control of Hyalomma ticks, vectors of the Crimean-Congo hemorrhagic fever virus: Where are we now and where are we going?

At a time of major global, societal, and environmental changes, the shifting distribution of pathogen vectors represents a real danger in certain regions of the world as generating opportunities for emergency. For example, the recent arrival of the Hyalomma marginatum ticks in southern France and the concurrent appearance of cases of Crimean-Congo hemorrhagic fever (CCHF)-a disease vectored by this tick species-in neighboring Spain raises many concerns about the associated risks for the European continent. This context has created an urgent need for effective methods for control, surveillance, and risk assessment for ticks and tick-borne diseases with a particular concern regarding Hyalomma sp. Here, we then review the current body of knowledge on different methods of tick control-including chemical, biological, genetical, immunological, and ecological methods-and the latest developments in the field, with a focus on those that have been tested against ticks from the genus Hyalomma. In the absence of a fully and unique efficient approach, we demonstrated that integrated pest management combining several approaches adapted to the local context and species is currently the best strategy for tick control together with a rational use of acaricide. Continued efforts are needed to develop and implement new and innovative methods of tick control.

Meteorological and climatic variables predict the phenology of Ixodes ricinus nymph activity in France, accounting for habitat heterogeneity.

Ixodes ricinus ticks (Acari: Ixodidae) are the most important vector for Lyme borreliosis in Europe. As climate change might affect their distributions and activities, this study aimed to determine the effects of environmental factors, i.e., meteorological, bioclimatic, and habitat characteristics on host-seeking (questing) activity of I. ricinus nymphs, an important stage in disease transmissions, across diverse climatic types in France over 8 years. Questing activity was observed using a repeated removal sampling with a cloth-dragging technique in 11 sampling sites from 7 tick observatories from 2014 to 2021 at approximately 1-month intervals, involving 631 sampling campaigns. Three phenological patterns were observed, potentially following a climatic gradient. The mixed-effects negative binomial regression revealed that observed nymph counts were driven by different interval-average meteorological variables, including 1-month moving average temperature, previous 3-to-6-month moving average temperature, and 6-month moving average minimum relative humidity. The interaction effects indicated that the phenology in colder climates peaked differently from that of warmer climates. Also, land cover characteristics that support the highest baseline abundance were moderate forest fragmentation with transition borders with agricultural areas. Finally, our model could potentially be used to predict seasonal human-tick exposure risks in France that could contribute to mitigating Lyme borreliosis risk.

Habitat suitability map of Ixodes ricinus tick in France using multi-criteria analysis.

The tick Ixodes ricinus is widely distributed across Europe and is responsible for the transmission of several pathogens to humans and animals. In this study, we used a knowledge-based method to map variations in habitat suitability for I. ricinus ticks throughout continental France and Corsica. The multi-criteria decision analysis (MCDA) integrated four major biotic and abiotic factors known to influence tick populations: climate, land cover, altitude and the density of wild ungulates. For each factor, habitat suitability index (HSI) values were attributed to different locations based on knowledge regarding its impact on tick populations. For the MCDA, two methods of factor combination were tested, additive and multiplicative, both which were evaluated at the spatial scales of departments and local municipalities. The resulting habitat suitability maps (resolution=100x100 m) revealed that conditions are suitable for I. ricinus over most of France and Corsica. Particularly suitable habitats were located in central, north-eastern and south-western France, while less-suitable habitats were found in the Mediterranean and mountainous regions. To validate the approach, the HSI scores were compared to field data of I. ricinus nymph abundance. Regardless of scale, the correlation between abundance indicator and HSI score was stronger for the additive than for the multiplicative approach. Overall, this study demonstrates the value of MCDA for estimating habitat suitability maps for I. ricinus abundance, which could be especially useful in highlighting areas of the tick's distribution where preventive measures should be prioritised.

Tick ecology and Lyme borreliosis prevention: a regional survey of pharmacists' knowledge in Auvergne-Rhône-Alpes, France.

The most prevalent vector-borne diseases in Europe are caused by tick-borne pathogens, such as bacteria of the genus Borrelia that cause Lyme borreliosis. In this context, retail pharmacists are frequently the first medical source of information in the event of a tick bite. The objective of this study was to assess pharmacy professionals' knowledge about both tick ecology and the appropriate measures for tick bites and Lyme borreliosis prevention. It was based on an online survey of 364 pharmacists and pharmacy assistants located in the Auvergne-Rhône-Alpes region of France. The results showed solid knowledge about preventive measures for tick bite and Lyme borreliosis, but weaker knowledge about tick biology (hosts, suitable habitats, favorable conditions for tick activity, etc.). In particular, several stereotypes were observed in the responses of the pharmacy professionals. These appear to result from a social construction of the knowledge on ticks and tick-borne diseases previously shown to the general population in the region. The results highlight the need for continuous training about ticks and tick-borne diseases for healthcare professionals serving local populations that live in endemic areas.

Exploring convolutional neural networks with transfer learning for diagnosing Lyme disease from skin lesion images.

BACKGROUND AND OBJECTIVE: Lyme disease which is one of the most common infectious vector-borne diseases manifests itself in most cases with erythema migrans (EM) skin lesions. Recent studies show that convolutional neural networks (CNNs) perform well to identify skin lesions from images. Lightweight CNN based pre-scanner applications for resource-constrained mobile devices can help users with early diagnosis of Lyme disease and prevent the transition to a severe late form thanks to appropriate antibiotic therapy. Also, resource-intensive CNN based robust computer applications can assist non-expert practitioners with an accurate diagnosis. The main objective of this study is to extensively analyze the effectiveness of CNNs for diagnosing Lyme disease from images and to find out the best CNN architectures considering resource constraints. METHODS: First, we created an EM dataset with the help of expert dermatologists from Clermont-Ferrand University Hospital Center of France. Second, we benchmarked this dataset for twenty-three CNN architectures customized from VGG, ResNet, DenseNet, MobileNet, Xception, NASNet, and EfficientNet architectures in terms of predictive performance, computational complexity, and statistical significance. Third, to improve the performance of the CNNs, we used custom transfer learning from ImageNet pre-trained models as well as pre-trained the CNNs with the skin lesion dataset HAM10000. Fourth, for model explainability, we utilized Gradient-weighted Class Activation Mapping to visualize the regions of input that are significant to the CNNs for making predictions. Fifth, we provided guidelines for model selection based on predictive performance and computational complexity. RESULTS: Customized ResNet50 architecture gave the best classification accuracy of 84.42% ±1.36, AUC of 0.9189±0.0115, precision of 83.1%±2.49, sensitivity of 87.93%±1.47, and specificity of 80.65%±3.59. A lightweight model customized from EfficientNetB0 also performed well with an accuracy of 83.13%±1.2, AUC of 0.9094±0.0129, precision of 82.83%±1.75, sensitivity of 85.21% ±3.91, and specificity of 80.89%±2.95. All the trained models are publicly available at https://dappem.limos.fr/download.html, which can be used by others for transfer learning and building pre-scanners for Lyme disease. CONCLUSION: Our study confirmed the effectiveness of even some lightweight CNNs for building Lyme disease pre-scanner mobile applications to assist people with an initial self-assessment and referring them to expert dermatologist for further diagnosis.

Questing tick abundance in urban and peri-urban parks in the French city of Lyon.

BACKGROUND: In Europe, ticks are responsible for the transmission of several pathogens of medical importance, including bacteria of the Borrelia burgdorferi (s.l.) complex, the agents of Lyme borreliosis. In France, the Auvergne Rhône-Alpes region is considered a hot spot for human tick-borne pathogen infections, with an estimated annual rate of 156 cases of Lyme borreliosis per 100,000 inhabitants. Although several studies have assessed the abundance of ticks in rural areas, little consideration has been given thus far to urban green spaces in France. METHODS: This study aimed to estimate tick abundance in three parks, two urban (U1, U2) and one peri-urban (PU), in and around the city of Lyon (France). A forest in a rural area was used as a control (C). Tick sampling campaigns were performed in each site in April, May, June, July, and October 2019 using the dragging method. One hundred transects of 10 m2 each were randomly chosen in each park in places frequented by humans. The sampling sessions were carried out under semi-controlled abiotic conditions. Ticks were stored in 70% ethanol and identified to species and developmental stage under a light microscope using morphological keys. RESULTS: A total of seven ticks (nymphs and adults) were collected in the two urban parks (six in U1 and one in U2), while 499 ticks were sampled in the peri-urban park. Of the 506 ticks collected, 504 were identified as Ixodes ricinus, one as Dermacentor marginatus, and one as Ixodes frontalis. In the peri-urban park, ticks were mainly collected under the forest cover and at forest edges. Tick density under forest cover was 7.1 times higher in the control site than in the peri-urban park throughout the survey period. CONCLUSIONS: This study confirmed the presence of ticks in all of the parks surveyed, although their occurrence in the urban parks was very rare compared to the peri-urban park and the control site. These results should serve as a basis for the implementation of preventive measures.

Distribution of ticks, tick-borne pathogens and the associated local environmental factors including small mammals and livestock, in two French agricultural sites: the OSCAR database.

BACKGROUND: In Europe, ticks are major vectors of both human and livestock pathogens (e.g. Lyme disease, granulocytic anaplasmosis, bovine babesiosis). Agricultural landscapes, where animal breeding is a major activity, constitute a mosaic of habitat types of various quality for tick survival and are used at different frequencies by wild and domestic hosts across seasons. This habitat heterogeneity, in time and space, conditions the dynamics of these host-vector-pathogen systems and thus drives acarological risk (defined as the density of infected ticks). The principal objective of the OSCAR project (2011-2016) was to examine the links between this heterogeneity and acarological risk for humans and their domestic animals. Here, we present the data associated with this project. NEW INFORMATION: This paper reports a database on the distribution and densities of I. ricinus ticks - the most common tick species in French agricultural landscapes - and the prevalence of three tick-borne pathogens (Anaplasma phagocytophilum, Borrelia spp. and Babesia spp.) in two sites in north-western ("Zone Atelier Armorique": ZA site) and south-western ("Vallées et Coteaux de Gascogne": VG site) France. The distribution and density of ticks along a gradient of wooded habitats, as well as biotic variables, such as the presence and abundance of their principal domestic (livestock) and wild hosts (small mammals), were measured from forest cores and edges to more or less isolated hedges, all bordering meadows. Ticks, small mammals and information on local environmental conditions were collected along 90 transects in each of the two sites in spring and autumn 2012 and 2013 and in spring 2014, corresponding to the main periods of tick activity. Local environmental conditions were recorded along each tick and small mammal transect: habitat type, vegetation type and characteristics, slope and traces of livestock presence. Samples consisted of questing ticks collected on the vegetation (mainly I. ricinus nymphs), biopsies of captured small mammals and ticks fixed on small mammals. In the VG site, livestock occurrence and abundance were recorded each week along each tick transect.A total of 29004 questing ticks and 1230 small mammals were captured during the study across the two sites and over the five field campaigns. All questing nymphs (N = 12287) and questing adults (N = 646) were identified to species. Ticks from small mammals (N = 1359) were also identified to life stage. Questing nymphs (N = 4518 I. ricinus) and trapped small mammals (N = 908) were analysed for three pathogenic agents: A. phagocytophilum, Borrelia spp. and Babesia spp.In the VG site, the average prevalence in I. ricinus nymphs for A. phagocytophilum, Borrelia spp. and Babesia spp. were, respectively 1.9% [95% CI: 1.2-2.5], 2.5% [95% CI: 1.8-3.2] and 2.7% [95% CI: 2.0-3.4]. In small mammals, no A. phagocytophilum was detected, but the prevalence for Borrelia spp. was 4.2% [95% CI: 0.9-7.5]. On this site, there was no screening of small mammals for Babesia spp. In ZA site, the average prevalence in nymphs for A. phagocytophilum, Borrelia spp. and Babesia were, respectively 2.2% [95% CI: 1.6-2.7], 3.0% [95% CI: 2.3-3.6] and 3.1% [95% CI: 2.5-3.8]. In small mammals, the prevalence of A. phagocytophilum and Borrelia spp. were, respectively 6.9% [95% CI: 4.9-8.9] and 4.1% [95% CI: 2.7-5.9]. A single animal was found positive for Babesia microti at this site amongst the 597 tested.

Rarity of microbial species: In search of reliable associations.

The role of microbial interactions in defining the properties of microbiota is a topic of key interest in microbial ecology. Microbiota contain hundreds to thousands of operational taxonomic units (OTUs), most of them rare. This feature of community structure can lead to methodological difficulties: simulations have shown that methods for detecting pairwise associations between OTUs, which presumably reflect interactions, yield problematic results. The performance of association detection tools is impaired when there is a high proportion of zeros in OTU tables. Our goal was to understand the impact of OTU rarity on the detection of associations. We explored the utility of common statistics for testing associations; the sensitivity of alternative association measures; and the performance of network inference tools. We found that a large proportion of pairwise associations, especially negative associations, cannot be reliably tested. This constraint could hamper the identification of candidate biological agents that could be used to control rare pathogens. Identifying testable associations could serve as an objective method for filtering datasets in lieu of current empirical approaches. This trimming strategy could significantly reduce the computational time needed to infer networks and network inference quality. Different possibilities for improving the analysis of associations within microbiota are discussed.

Using design theory to foster innovative cross-disciplinary research: Lessons learned from a research network focused on antimicrobial use and animal microbes' resistance to antimicrobials.

Dealing with the major societal and research challenges related to antimicrobial use will require cross-disciplinary research and strong relationships between researchers and stakeholders. Design theories, such as the concept-knowledge (C-K) theory, can help spur the emergence of innovation. Here, our objective was to examine how the C-K theory could promote the development of novel, cross-disciplinary research projects on antimicrobial use and animal microbes' resistance to antimicrobials. A French research network (R2A2; Réseau Recherche Antibiotiques Animal) was created whose goal was to foster cross-disciplinary research and scientific discussion on these topics. The R2A2 network hosted general meetings and thematic workshops, during which participants brainstormed using C-K diagrams. The network's performance was evaluated through the evolution of C-K diagrams, project creation, and participant interviews. R2A2 led to the creation of a minimum of eight research projects. The participants felt network events facilitated interactions and collaborations with researchers in different disciplines. The R2A2 network has opened new avenues of research into several important topics: antimicrobial use on farms, the environmental impacts of antimicrobials, animal immunity, and alternative treatments. The keys to its success were: (i) participant interest; (ii) the use of C-K design theory to encourage cross-disciplinary thinking; (iii) the aim of fostering several small projects rather than one large project; and (iv) network responsiveness to participant needs with regards to meeting and workshop topics. C-K theory served a key role in promoting cross-disciplinary thinking on topics at the interface between research and stakeholder interests.

Population genetics, community of parasites, and resistance to rodenticides in an urban brown rat (Rattus norvegicus) population.

Brown rats are one of the most widespread urban species worldwide. Despite the nuisances they induce and their potential role as a zoonotic reservoir, knowledge on urban rat populations remains scarce. The main purpose of this study was to characterize an urban brown rat population from Chanteraines park (Hauts-de-Seine, France), with regards to haematology, population genetics, immunogenic diversity, resistance to anticoagulant rodenticides, and community of parasites. Haematological parameters were measured. Population genetics was investigated using 13 unlinked microsatellite loci. Immunogenic diversity was assessed for Mhc-Drb. Frequency of the Y139F mutation (conferring resistance to rodenticides) and two linked microsatellites were studied, concurrently with the presence of anticoagulant residues in the liver. Combination of microscopy and molecular methods were used to investigate the occurrence of 25 parasites. Statistical approaches were used to explore multiple parasite relationships and model parasite occurrence. Eighty-six rats were caught. The first haematological data for a wild urban R. norvegicus population was reported. Genetic results suggested high genetic diversity and connectivity between Chanteraines rats and surrounding population(s). We found a high prevalence (55.8%) of the mutation Y139F and presence of rodenticide residues in 47.7% of the sampled individuals. The parasite species richness was high (16). Seven potential zoonotic pathogens were identified, together with a surprisingly high diversity of Leptospira species (4). Chanteraines rat population is not closed, allowing gene flow and making eradication programs challenging, particularly because rodenticide resistance is highly prevalent. Parasitological results showed that co-infection is more a rule than an exception. Furthermore, the presence of several potential zoonotic pathogens, of which four Leptospira species, in this urban rat population raised its role in the maintenance and spread of these pathogens. Our findings should stimulate future discussions about the development of a long-term rat-control management program in Chanteraines urban park.

Ecological factors influencing small mammal infection by Anaplasma phagocytophilum and Borrelia burgdorferi s.l. in agricultural and forest landscapes.

Small mammals are key components of numerous tick-borne disease systems, as hosts for immature ticks and pathogen reservoirs. To study the factors influencing tick-borne infection in small mammals, we trapped small mammals and collected questing ticks in spring and autumn in 2012 and 2013 at 24 sites in a 10 × 15 km rural landscapes (Brittany, France). Tissue samples were screened by real-time PCR for Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato. Of the two dominant small mammal species captured, bank voles (Myodes glareolus) had higher prevalence than wood mice (Apodemus sylvaticus) for both infections, presumably because of specific differences in immunological defenses. Prevalence of infections was higher in 2013 than in 2012, likely because small mammals were fivefold less abundant in 2013, favouring tick aggregation. Bacterial prevalence, which was higher in autumn, was not associated to questing Ixodes ricinus nymph abundance which was six times higher in spring, but rather to the structure of the small mammal community. These findings suggest the involvement of endophilic tick species, I. trianguliceps and/or I. acuminatus, in bacterial transmission. Our study highlights that the entire community of hosts and vectors, and their interactions, should be considered to fully understand the epidemiology of vector-borne diseases.

Host specificity, pathogen exposure, and superinfections impact the distribution of Anaplasma phagocytophilum genotypes in ticks, roe deer, and livestock in a fragmented agricultural landscape.

Anaplasma phagocytophilum is a bacterial pathogen mainly transmitted by Ixodes ricinus ticks in Europe. It infects wild mammals, livestock, and, occasionally, humans. Roe deer are considered to be the major reservoir, but the genotypes they carry differ from those that are found in livestock and humans. Here, we investigated whether roe deer were the main source of the A. phagocytophilum genotypes circulating in questing I. ricinus nymphs in a fragmented agricultural landscape in France. First, we assessed pathogen prevalence in 1837 I. ricinus nymphs (sampled along georeferenced transects) and 79 roe deer. Prevalence was dramatically different between ticks and roe deer: 1.9% versus 76%, respectively. Second, using high-throughput amplicon sequencing, we characterized the diversity of the A. phagocytophilum genotypes found in 22 infected ticks and 60 infected roe deer; the aim was to determine the frequency of co-infections. Only 22.7% of infected ticks carried genotypes associated with roe deer. This finding fits with others suggesting that cattle density is the major factor explaining infected tick density. To explore epidemiological scenarios capable of explaining these patterns, we constructed compartmental models that focused on how A. phagocytophilum exposure and infection dynamics affected pathogen prevalence in roe deer. At the exposure levels predicted by the results of this study and the literature, the high prevalence in roe deer was only seen in the model in which superinfections could occur during all infection phases and when the probability of infection post exposure was above 0.43. We then interpreted these results from the perspective of livestock and human health.

Hedgehogs and Mustelid Species: Major Carriers of Pathogenic Leptospira, a Survey in 28 Animal Species in France (20122015).

Human leptospirosis is a zoonotic and potentially fatal disease that has increasingly been reported in both developing and developed countries, including France. However, our understanding of the basic aspects of the epidemiology of this disease, including the source of Leptospira serogroup Australis infections in humans and domestic animals, remains incomplete. We investigated the genetic diversity of Leptospira in 28 species of wildlife other than rats using variable number tandem repeat (VNTR) and multispacer sequence typing (MST). The DNA of pathogenic Leptospira was detected in the kidney tissues of 201 individuals out of 3,738 tested individuals. A wide diversity, including 50 VNTR profiles and 8 MST profiles, was observed. Hedgehogs and mustelid species had the highest risk of being infected (logistic regression, OR = 66.8, CI95% = 30.9-144 and OR = 16.7, CI95% = 8.7-31.8, respectively). Almost all genetic profiles obtained from the hedgehogs were related to Leptospira interrogans Australis, suggesting the latter as a host-adapted bacterium, whereas mustelid species were infected by various genotypes, suggesting their interaction with Leptospira was different. By providing an inventory of the circulating strains of Leptospira and by pointing to hedgehogs as a potential reservoir of L. interrogans Australis, our study advances current knowledge on Leptospira animal carriers, and this information could serve to enhance epidemiological investigations in the future.

Strong evidence for the presence of the tick Hyalomma marginatum Koch, 1844 in southern continental France.

Hyalomma ticks can transmit several human and animal pathogens in Eurasia and Africa. Interest in Hyalomma marginatum has increased since the recent (re)emergence of Crimean-Congo Hemorrhagic fever in the Palearctic region. Until now, continental France has been considered free of this tick species. Nevertheless, the existence of incomplete and occasionally incorrect records has maintained confusion about its status. Based on several tick sampling campaigns conducted on horses and birds from 2007 to 2016, we provided very strong evidence for the presence of reproducing populations of H. marginatum in parts of southern continental France. We also confirmed the introduction of immature developmental stages of H. marginatum, as well as H. rufipes, into France probably through trans-Mediterranean bird migrations.

Multiple independent transmission cycles of a tick-borne pathogen within a local host community.

Many pathogens are maintained by multiple host species and involve multiple strains with potentially different phenotypic characteristics. Disentangling transmission patterns in such systems is often challenging, yet investigating how different host species contribute to transmission is crucial to properly assess and manage disease risk. We aim to reveal transmission cycles of bacteria within the Borrelia burgdorferi species complex, which include Lyme disease agents. We characterized Borrelia genotypes found in 488 infected Ixodes ricinus nymphs collected in the Sénart Forest located near Paris (France). These genotypes were compared to those observed in three sympatric species of small mammals and network analyses reveal four independent transmission cycles. Statistical modelling shows that two cycles involving chipmunks, an introduced species, and non-sampled host species such as birds, are responsible for the majority of tick infections. In contrast, the cycle involving native bank voles only accounts for a small proportion of infected ticks. Genotypes associated with the two primary transmission cycles were isolated from Lyme disease patients, confirming the epidemiological threat posed by these strains. Our work demonstrates that combining high-throughput sequence typing with networks tools and statistical modeling is a promising approach for characterizing transmission cycles of multi-host pathogens in complex ecological settings.

Discovery of hantavirus circulating among Rattus rattus in French Mayotte island, Indian Ocean.

Hantaviruses are emerging zoonotic viruses that cause human diseases. In this study, sera from 642 mammals from La Réunion and Mayotte islands (Indian Ocean) were screened for the presence of hantaviruses by molecular analysis. None of the mammals from La Réunion island was positive, but hantavirus genomic RNA was discovered in 29/160 (18 %) Rattus rattus from Mayotte island. The nucleoprotein coding region was sequenced from the liver and spleen of all positive individuals allowing epidemiological and intra-strain variability analyses. Phylogenetic analysis based on complete coding genomic sequences showed that this Murinae-associated hantavirus is a new variant of Thailand virus. Further studies are needed to investigate hantaviruses in rodent hosts and in Haemorrhagic Fever with Renal Syndrome (HFRS) human cases.

Co-infection of Ticks: The Rule Rather Than the Exception.

INTRODUCTION: Ticks are the most common arthropod vectors of both human and animal diseases in Europe, and the Ixodes ricinus tick species is able to transmit a large number of bacteria, viruses and parasites. Ticks may also be co-infected with several pathogens, with a subsequent high likelihood of co-transmission to humans or animals. However few data exist regarding co-infection prevalences, and these studies only focus on certain well-known pathogens. In addition to pathogens, ticks also carry symbionts that may play important roles in tick biology, and could interfere with pathogen maintenance and transmission. In this study we evaluated the prevalence of 38 pathogens and four symbionts and their co-infection levels as well as possible interactions between pathogens, or between pathogens and symbionts. METHODOLOGY/PRINCIPAL FINDINGS: A total of 267 Ixodes ricinus female specimens were collected in the French Ardennes and analyzed by high-throughput real-time PCR for the presence of 37 pathogens (bacteria and parasites), by rRT-PCR to detect the presence of Tick-Borne encephalitis virus (TBEV) and by nested PCR to detect four symbionts. Possible multipartite interactions between pathogens, or between pathogens and symbionts were statistically evaluated. Among the infected ticks, 45% were co-infected, and carried up to five different pathogens. When adding symbiont prevalences, all ticks were infected by at least one microorganism, and up to eight microorganisms were identified in the same tick. When considering possible interactions between pathogens, the results suggested a strong association between Borrelia garinii and B. afzelii, whereas there were no significant interactions between symbionts and pathogens. CONCLUSION/SIGNIFICANCE: Our study reveals high pathogen co-infection rates in ticks, raising questions about possible co-transmission of these agents to humans or animals, and their consequences to human and animal health. We also demonstrated high prevalence rates of symbionts co-existing with pathogens, opening new avenues of enquiry regarding their effects on pathogen transmission and vector competence.