Intra- and interspecies infectious neighbourhoods as determinant parameters for Mycobacterium bovis infection among badgers in southwestern France.

The epidemiological system for Mycobacterium bovis in France involves cattle and, in some areas, wildlife species (mainly badgers and wild boar). This multi-host aspect complicates the control and eradication prospects for bovine tuberculosis in endemic areas, despite the surveillance and control measures implemented for decades in this officially tuberculosis-free European country. To improve control measures, and to manage spillback transmission from badgers to cattle, it is necessary to clarify the transmission mechanisms of M. bovis in these epidemiological systems. We modelled a badger population from a southwestern endemic area by a Dirichlet tessellation based on a sett census conducted by local hunters and trappers between 2013 and 2015. We then used a logistic regression model to test the association between the infection status of setts and computed variables depicting three types of transmission (intraspecific, interspecific and landscape-associated). The apparent prevalence of infected setts was of 40.5%. Two variables were significantly associated with the probability for a sett to be infected: the proportion of neighbouring setts that were infected (OR: 3.19 [2.04-5.17]95%) and the presence of nearby pastures belonging to an infected farm (OR: 2.33 [1.13-4.89]95%]. While badger culling measures have been implemented according to the national TB control plan in the study area since 2012 (in the vicinity of infected farms and their pastures), our results clearly highlight the need to reinforce measures aimed at reducing both intraspecific and interspecific infection pressure. For this purpose, the promising prospect of badger vaccination could be considered, along with biosecurity measures.

Spatial association of Mycobacterium bovis infection in cattle and badgers at the pasture interface in an endemic area in France.

Despite control and surveillance programmes, Mycobacterium bovis, the main aetiologic agent of bovine tuberculosis (bTB), is still detected on cattle farms and in wildlife populations in France, especially in badgers in the French Côte-d'Or département. The aim of our study was to find out if infected badgers were trapped significantly closer to pastures of infected farms than non-infected badgers and, if so, to determine the most efficient distance around those pastures for badger trapping, particularly for surveillance purposes. We studied two subareas (southern and northern), chosen based on natural barriers to badger movements and according to the presence of pastures belonging to infected farms (POIFs) and infected or non-infected badgers. In each subarea, we computed the shortest distances D0 and D between badgers trapped a given year n between 2015 and 2019 (n = 59 infected and n = 1535 non-infected badgers for D0; n = 53 infected and n = 1476 non-infected badgers for D) and POIFs designated as infected between the year n - 4 and n + 1 (respectively n = 373 and n = 388 POIFs). D0 was calculated without considering spoligotypes, while D was calculated considering the possible epidemiological link between infected badgers and POIFs by using bTB spoligotype information. Then, we computed the observed mean and median of the D0 and D distances and used a bootstrap analysis to test if infected badgers were found significantly closer to POIFs than non-infected badgers. We observed that infection of badgers was not independent of distance from POIF in both subareas but distances (D0 or D) were different between the northern and southern subarea. In the northern subarea, which displays a mosaic landscape (mean and median D distances were respectively 612 m and 303 m for infected badgers), infected badgers indeed were trapped closer to POIFs, considering D0 and D. In the southern subarea, predominantly forested, infected badgers were significantly closer to POIFs than non-infected badgers when considering D0 but not for D (mean and median D distances were respectively 7148 m and 4831 m for infected badgers). These results will help to determine the most efficient distance from POIFs to trap badgers to determine their infection status in countryside landscapes. They also highlight the need to better understand the epidemiological systems at play in more forested landscapes where badgers may behave differently or other susceptible sympatric wild species might play a more important role in the circulation of M. bovis, both phenomena contributing to badger infection at greater distances from POIFs.

"BACACIX", a spatial index combining proxies of bovine and badger space use associated with extended Mycobacterium bovis circulation in France.

To better prevent and control multi-host pathogen circulation over large areas, it is essential to identify patterns of disease persistence within host communities involved in pathogen circulation at a macroscale. The aim of this study was to design and calculate "BACACIX", a spatial index of indirect contacts between cattle and badgers, two species involved in the circulation of Mycobacterium bovis, one of the main causative agents of bovine tuberculosis (bTB), in some areas of France. The index combined spatial models of land use distribution (the probable distribution defining animal use of space) based on pasture location for cattle, and based on land cover for badgers, with proxies for animal density for both species. For badgers, we used two series of census data of badger setts in two regions of France to evaluate our model of badger space use distribution (also known as utilization distribution), and analyzed the relationship between BACACIX and the upsurge of bovine tuberculosis observed in several regions of France during the decade after the country obtained the officially bTB-free status in 2001. We observed high values of BACACIX from the southwest to the northeast of France and from Brittany to the Channel coast. Conversely, in two areas (north-central area and Mediterranean coast), index values were low, suggesting that indirect cattle-badger contacts were unlikely. In the two series of census data of badger setts that we analyzed, 96.5% and 87% of the global positioning system (GPS) locations of badger setts, respectively were located in the calculated badger space use distribution. A logistic regression model showed that after controlling bTB over the previous decade, the value of the index was positively associated with the risk of cattle outbreaks between 2001 and 2010 (OR = 1.57). In addition, the risk of bTB occurrence in cattle decreased when the pasture area outside the badger space use distribution increased. In the future, the spatial index of indirect cattle-badger contacts we propose could help to better target bTB surveillance and control in France.

Predicting veal-calf trading events in France.

Global trade has been ranked as one of the top five drivers of infectious disease threat events. More specifically, livestock trade is known to increase the speed at which infectious diseases circulate and to facilitate their dissemination over large distances Therefore, predicting animal movements arising from trade is crucial for assessing epidemic risk and the impact of preventive measures. In this study, we developed a statistical framework for predicting trading events using predictors accessible from routinely collected data. We focused on veal calves, a category of animals with significant commercial value; the dataset considered the veal calf trade in France between January 2011 and June 2019. A subset of farms with consistent trade behaviour over time was built to be used throughout the study. To predict sale or purchase event occurrences, our predictive framework was based on random forests as a binary classification tool, an approach that allows a large number of potential predictors. We explored the robustness of model predictions with respect to the delay in data acquisition and prediction lag time. Overall, sales were more accurately predicted than purchasing events. Unsurprisingly, a delay in data acquisition led to a decrease in the performance of indicators, whereas prediction lag time had little impact. Sale-related predictors mostly reflected past trading events, whereas purchase-related predictors were associated with past trading events, farm management and general farm characteristics. The model outputs also suggested that the veal calf trading network is driven by sales rather than by purchases. Regardless of the length of the delay in data acquisition and prediction lag, the random forest approach fitted on data with municipality as trading unit and a 28-day trading period provided better performance scores (F1-score, positive predictive value and negative predictive value) than scenarios with finer temporal and spatial aggregation units. Predicted trade events can therefore be used to reconstruct the entire veal calf trading network and transfers between selling and purchasing units for each period. This predicted network could be further used to simulate the spread of pathogens via animal trade.

Diversity of Tick-Borne Pathogens in Tick Larvae Feeding on Breeding Birds in France.

Birds play a role in maintaining tick-borne diseases by contributing to the multiplication of ticks and pathogens on a local scale during the breeding season. In the present study, we describe the diversity of tick and pathogen species of medical and veterinary importance in Europe hosted by 1040 captured birds (56 species) during their breeding season in France. Of the 3114 ticks collected, Ixodes ricinus was the most prevalent species (89.5%), followed by I. frontalis (0.8%), I. arboricola (0.7%), Haemaphysalis concinna (0.5%), H. punctata (0.5%), Hyalomma spp. (0.2%), and Rhipicephalus spp. (0.06%). Because they may be representative of the bird infection status for some pathogen species, 1106 engorged tick larvae were screened for pathogens. Borrelia burgdorferi sensu lato was the most prevalent pathogen genus in bird-feeding larvae (11.7%), followed by Rickettsia spp. (7.4%), Anaplasma spp. (5.7%), Babesia spp. (2.3%), Ehrlichia spp. (1.4%), and B. miyamotoi (1%). Turdidae birds (Turdus merula and T. philomelos), Troglodytes troglodytes, and Anthus trivialis had a significantly higher prevalence of B. burgdorferi s.l.-infected larvae than other pathogen genera. This suggests that these bird species could act as reservoir hosts for B. burgdorferi s.l. during their breeding season, and thus play an important role in acarological risk.

How far away do you keep your equines? Estimation of the equine population's spatial distribution in France.

It is essential to have an accurate picture of the spatial distribution of equines to be able to monitor equine health events effectively. In France, this information is only available for certain categories of live equines kept in professional structures and for dead equines removed by renderers. This limits the surveillance, prevention and control methods able to be used to prevent the spread of equine diseases. Our study aimed to provide a realistic estimate of the spatial distribution of the French equine population at the detailed scale of the French commune (France's smallest administrative unit). For this purpose, we adapted the Bayesian method used by Lo Lacono et al., based on the distance between the owner's location and the location of his/her equines, and on the percentage of urban coverage. To assess whether the location of dead equines could be representative of the location of live equines, the distribution of distances between equines and owners was calculated from a sample of live equines on the one hand, and a sample of dead equines on the other, both accurately located. We also tested two different assignment methods for locating equines: Method 1 assigned to each owner a single holding commune, while Method 2 allowed more variability in holding communes for owners associated with multiple equines. A marked difference was observed between Methods 1 and 2 regardless of the sample used, with only 2.4% and 4.3% respectively of the communes having the same number of equines. Conversely, little difference was observed in the results whether the live or dead equine sample was used, with approximately 45% of the communes having the same number of equines. Regarding differences in spatial distribution, Method 1 based on the live equine sample estimated higher local densities of equines without considering urban areas. In contrast, Method 2 provided more dispersed maps, with low densities in the densest urban areas. In conclusion, dead equines appeared to be representative of live equines and some of our estimates are consistent with the information collected by the French horse and riding institute (IFCE). These results now have to be compared with field data to test their relevance.

Spatial and Temporal Circulation of Babesia caballi and Theileria equi in France Based on Seven Years of Serological Data.

Caused by two blood parasites, Babesia caballi and Theileria equi, equine piroplasmosis is a tick-borne disease that poses major health and economic issues for the equine industry. Our objective was to gain insight into the spatio-temporal variations of parasite circulation in France, where the disease is known to be enzootic, but has been the subject of few studies. Seroprevalence was assessed for each parasite thanks to 16,127 equine sera obtained between 1997 and 2003 from all over France and analysed through complement fixation tests. Results indicated that 13.2% (5-27% depending on the region) of horses were seropositive for T. equi and 9.5% (3-25%) for B. caballi. Regardless of the year, horses from the southern regions of France were the most affected by B. caballi or T. equi infection, while the proportion of horses having antibodies against T. equi increased over time. These results highlight the heterogeneity of the circulation of both piroplasms, which may be linked with ecological diversity and vector distribution. Our data provide baseline information regarding the sero-epidemiology of B. caballi and T. equi infection in horses in France, making it now possible to select regions for future studies on risk factors, and design and implement effective targeted measures against equine piroplasms.

Research effort on birds' reservoir host potential for Lyme borreliosis: A systematic review and perspectives.

Zoonotic tick-borne diseases threat human and animal health. Understanding the role of hosts in the production of infected ticks in an epidemiological system is essential to be able to design effective measures to reduce the exposure of humans and animals to infectious tick bites. The reservoir host potential, that is, number of infected ticks produced by a host species, depends on three components: tick production, realized reservoir competence and host density. The parameters and factors that determine the reservoir host potential need to be characterized to achieve a robust understanding of the dynamics of pathogen-tick-host systems, and thus to mitigate the acarological risk of emerging infections. Few studies have investigated the role of birds in the local spread of Lyme borreliosis Borrelia. Knowledge of the research effort on the reservoir host potential of birds in Lyme borreliosis Borrelia circulation is necessary to prioritize future research on this topic. We provide a systematic review of the research effort on components of the reservoir host potential of wild birds for Lyme borreliosis Borrelia circulation, and factors that modulate these components in the European epidemiological system. Our review of 242 selected publications showed that tick production has been 1.4 and 21 times more studied than realized reservoir competence and bird density respectively. Only one study achieved to characterize the global host reservoir potential of birds in a given epidemiological system. Investigated factors were mostly related to bird species identity, individual characteristics of birds and tick characteristics, whereas the influence of bird life-history traits have been largely under-investigated. Because simultaneous characterization of all parameters is notoriously complex, interdisciplinary research is needed to combine and accumulate independent field and laboratory investigations targeting each parameter on specific epidemiological system or host species. This can help gain an integrated appraisal of the functioning of the studied system at a local scale.

Eco-epidemiology of equine piroplasmosis and its associated tick vectors in Europe: A systematic literature review and a meta-analysis of prevalence.

When studying a vector-borne disease, an eco-epidemiological approach is vital for a comprehensive understanding of how the pathogen circulates amongst populations. Equine piroplasmosis (EP), a tick-borne disease caused by the protozoans Babesia caballi and Theileria equi, is endemic in the Mediterranean basin of Europe and causes both animal health and economic issues for the equine sector. With no vaccine available, defining the episystem of the disease can help to identify which components of the host-pathogen-vector-environment system to target to improve preventive measures. In this systematic literature review, we collected relevant data on the eco-epidemiology of EP in Europe. The 62 studies remaining after the selection procedure explored potential vectors, indicators of parasite circulation and putative risk factors of EP. Eight hard tick species were identified as potential vectors of one or both piroplasm species. Meta-analyses were then conducted on prevalence and seroprevalence data in equids in European countries, demonstrating an estimated seroprevalence of 30% and 8% and prevalence of 25% and 2% for T. equi and B. caballi, respectively. Finally, herd management practices and environmental risk factors analysed in studies showed no real consensus between studies, but revealed a general trend highlighting age and exposure to ticks as risk factors, and vaccination as a protective factor. Through this study, we point out that only a few studies have focused on disease management practices and even fewer have studied the effect of environmental parameters on equid infections. Further investigation in these areas is required to better characterize the eco-epidemiology of EP and risk factors associated with this disease.

Evaluation of human resources needed and comparison with human resources available to implement emergency vaccination in case of foot and mouth disease outbreaks in Tunisia.

Foot and mouth disease (FMD) is a highly contagious viral disease that affects domestic and wild artiodactyl animals and causes considerable economic losses related to outbreak management, production losses and trade impacts. In Tunisia, the last FMD outbreak took place in 2018-2019. The effectiveness of control measures implemented to control FMD depends, in particular, on the human resources used to implement them. Tunisia has the ultimate objective of obtaining OIE status as 'FMD-free with vaccination'. The aim of this study was to determine and compare the necessary and available human resources to control FMD outbreaks in Tunisia using emergency vaccination and to assess the gaps that would play a role in the implementation of the strategy. We developed a resources-requirement grid of necessary human resources for the management of the emergency vaccination campaign launched after the identification of a FMD-infected premises in Tunisia. Field surveys, conducted in the 24 governorates of Tunisia, allowed quantifying the available human resources for several categories of skills considered in the resources-requirement grid. For each governorate, we then compared available and necessary human resources to implement vaccination according to eight scenarios mixing generalised or cattle-targeted vaccination and different levels of human resources. The resources-requirement grid included 11 tasks in three groups: management of FMD-infected premises, organisational tasks and vaccination implementation. The available human resources for vaccination-related tasks included veterinarians and technicians from the public sector and appointed private veterinarians. The comparison of available and necessary human resources showed vaccination-related tasks to be the most time-consuming in terms of managing a FMD outbreak. Increasing the available human resources using appointed private veterinarians allowed performing the emergency vaccination of animals in the governorate in due time, especially if vaccination was targeted on cattle. The overall approach was validated by comparing the predicted and observed durations of a vaccination campaign conducted under the same conditions as during the 2014 Tunisian outbreak. This study could provide support to the Tunisian Veterinary Services or to other countries to optimise the management of a FMD outbreak.

Taxon Appearance From Extraction and Amplification Steps Demonstrates the Value of Multiple Controls in Tick Microbiota Analysis.

BACKGROUND: The development of high-throughput sequencing technologies has substantially improved analysis of bacterial community diversity, composition, and functions. Over the last decade, high-throughput sequencing has been used extensively to identify the diversity and composition of tick microbial communities. However, a growing number of studies are warning about the impact of contamination brought along the different steps of the analytical process, from DNA extraction to amplification. In low biomass samples, e.g., individual tick samples, these contaminants may represent a large part of the obtained sequences, and thus generate considerable errors in downstream analyses and in the interpretation of results. Most studies of tick microbiota either do not mention the inclusion of controls during the DNA extraction or amplification steps, or consider the lack of an electrophoresis signal as an absence of contamination. In this context, we aimed to assess the proportion of contaminant sequences resulting from these steps. We analyzed the microbiota of individual Ixodes ricinus ticks by including several categories of controls throughout the analytical process: homogenization, DNA extraction, and DNA amplification. RESULTS: Controls yielded a significant number of sequences (1,126-13,198 mean sequences, depending on the control category). Some operational taxonomic units (OTUs) detected in these controls belong to genera reported in previous tick microbiota studies. In this study, these OTUs accounted for 50.9% of the total number of sequences in our samples, and were considered contaminants. Contamination levels (i.e., the percentage of sequences belonging to OTUs identified as contaminants) varied with tick instar and sex: 76.3% of nymphs and 75% of males demonstrated contamination over 50%, while most females (65.7%) had rates lower than 20%. Contamination mainly corresponded to OTUs detected in homogenization and extraction reagent controls, highlighting the importance of carefully controlling these steps. CONCLUSION: Here, we showed that contaminant OTUs from sample laboratory processing steps can represent more than half the total sequence yield in sequencing runs, and lead to unreliable results when characterizing tick microbial communities. We thus strongly advise the routine use of negative controls in tick microbiota studies, and more generally in studies involving low biomass samples.

Serological Evidence of West Nile and Usutu Viruses Circulation in Domestic and Wild Birds in Wetlands of Mali and Madagascar in 2008.

The geographical distribution and impact on animal and human health of both West Nile and Usutu viruses, two flaviviruses of the Japanese encephalitis complex, have been increasing during the past two decades. Both viruses circulate in Europe and Africa within a natural cycle between wild birds and mosquitoes, mainly from the Culex genus. We retrospectively analyzed sera from domestic and wild birds sampled in 2008 in two wetlands, namely the Inner Niger Delta, Mali, and the Lake Alaotra area, Madagascar. Sera were first tested using a commercial ID Screen West Nile Competition Multi-species ELISA kit. Then, positive sera and sera with insufficient volume for testing with ELISA were tested with a Microneutralization Test. In Mali, the observed seroprevalence in domestic birds was 28.5% [24.5; 32.8] 95%CI, 3.1 % [1.8; 5.2] 95%CI, 6.2% [3.4; 10.2] 95%CI and 9.8 % [7.3; 12.8] 95%CI, for West Nile virus (WNV), Usutu virus (USUV), undetermined flavivirus, and WNV/USUV respectively. Regarding domestic birds of Madagascar, the observed seroprevalence was 4.4 % [2.1; 7.9]95%CI for WNV, 0.9% [0.1; 3.1] 95%CI for USUV, 1.3% [0.5; 2.8] 95%CI for undetermined flavivirus, and null for WNV/USUV. Among the 150 wild birds sampled in Madagascar, two fulvous whistling-ducks (Dendrocygna bicolor) were positive for WNV and two for an undetermined flavivirus. One white-faced whistling-duck (Dendrocygna viduata) and one Hottentot teal (Spatula hottentota) were tested positive for USUV. African and European wetlands are linked by wild bird migrations. This first detection of USUV-as well as the confirmed circulation of WNV in domestic birds of two wetlands of Mali and Madagascar-emphasizes the need to improve the surveillance, knowledge of epidemiological patterns, and phylogenetic characteristics of flavivirus in Africa, particularly in areas prone to sustained, intense flavivirus transmission such as wetlands.

A three-years assessment of Ixodes ricinus-borne pathogens in a French peri-urban forest.

BACKGROUND: Ixodes ricinus is the predominant tick species in Europe and the primary pathogen vector for both humans and animals. These ticks are frequently involved in the transmission of Borrelia burgdorferi (sensu lato), the causative agents of Lyme borreliosis. While much more is known about I. ricinus tick-borne pathogen composition, information about temporal tick-borne pathogen patterns remain scarce. These data are crucial for predicting seasonal/annual patterns which could improve understanding and prevent tick-borne diseases. METHODS: We examined tick-borne pathogen (TBP) dynamics in I. ricinus collected monthly in a peri-urban forest over three consecutive years. In total, 998 nymphs were screened for 31 pathogenic species using high-throughput microfluidic real-time PCR. RESULTS: We detected DNA from Anaplasma phagocytophilum (5.3%), Rickettsia helvetica (4.5%), Borrelia burgdorferi (s.l.) (3.7%), Borrelia miyamotoi (1.2%), Babesia venatorum (1.5%) and Rickettsia felis (0.1%). Among all analysed ticks, 15.9% were infected by at least one of these microorganisms, and 1.3% were co-infected. Co-infections with B. afzeli/B. garinii and B. garinii/B. spielmanii were significantly over-represented. Moreover, significant variations in seasonal and/or inter-annual prevalence were observed for several pathogens (R. helvetica, B. burgdorferi (s.l.), B. miyamotoi and A. phagocytophilum). CONCLUSIONS: Analysing TBP prevalence in monthly sampled tick over three years allowed us to assess seasonal and inter-annual fluctuations of the prevalence of TBPs known to circulate in the sampled area, but also to detect less common species. All these data emphasize that sporadic tick samplings are not sufficient to determine TBP prevalence and that regular monitoring is necessary.

The Immunomodulatory Effect of IrSPI, a Tick Salivary Gland Serine Protease Inhibitor Involved in Ixodes ricinus Tick Feeding.

Ticks are the most important vectors of pathogens affecting both domestic and wild animals worldwide. Hard tick feeding is a slow process-taking up to several days-and necessitates extended control over the host response. The success of the feeding process depends upon injection of tick saliva, which not only controls host hemostasis and wound healing, but also subverts the host immune response to avoid tick rejection that creates a favorable niche for the survival and propagation of diverse tick-borne pathogens. Here, we report on the molecular and biochemical features and functions of an Ixodes ricinus serine protease inhibitor (IrSPI). We characterize IrSPI as a Kunitz elastase inhibitor that is overexpressed in several tick organs-especially salivary glands-during blood-feeding. We also demonstrated that when IrSPI is injected into the host through saliva, it had no impact on tissue factor pathway-induced coagulation, fibrinolysis, endothelial cell angiogenesis or apoptosis, but the protein exhibits immunomodulatory activity. In particular, IrSPI represses proliferation of CD4+ T lymphocytes and proinflammatory cytokine secretion from both splenocytes and macrophages. Our study contributes valuable knowledge to tick-host interactions and provides insights that could be further exploited to design anti-tick vaccines targeting this immunomodulator implicated in I. ricinus feeding.

Anterior segment parameters measured in young healthy cats using a rotating Scheimpflug camera.

OBJECTIVE: The purpose of this study is to measure parameters of the anterior segment of normal feline eyes using a rotating Scheimpflug camera in young and adult cats. PROCEDURE: Sixteen domestic short-haired (DSH) healthy cats (seven castrated males and nine spayed females) with a mean age of 17 months (SD = 3.5) were anesthetized. Cats were considered immature and adults if they were < or ≥12 months of age, respectively. The keratometric values, the anterior chamber depth (ACD), and central corneal thickness (CCT) of the right eye of each animal were measured using a Pentacam Oculus topographer. The repeatability of the measures was assessed by calculating the coefficient of variation of the successive measures in cats (five repetitions per cat). We performed an ANOVA to test the effect of age on the parameters measures. RESULTS: The mean CCT and ACD were 601.97 ± 38 µm and 5.27 ± 0.04 mm, respectively. No significant differences were observed between adult and young cat's measures. The average keratometric value was 39.6 ± 0.3 D. The mean average cylindrical error was 2.48 ± 0.35 D (43.75% "against-the-rule" astigmatism and 43.75% "oblique" astigmatism). CONCLUSION: The Oculus Pentacam under clinical conditions give CCT and ACD values very similar to those reported in previous studies. Corneal astigmatism measures were markedly elevated (>2 D). Further studies with a larger population of cats of various breeds and ages are needed to confirm or refute these results, using a rotating Scheimpflug camera.

Characterisation of the French ferret population, husbandry, reported medical care and feeding habits.

Ferrets have become a common companion animal. But no data are available on the French population of ferrets. The goal of the survey was to characterise this population, including demographic characteristics, husbandry, reported medical care and feeding habits. Complete data were available for 1205 pet ferrets in 709 households. Most ferrets (86·1 %) lived indoors, 1 % received only artificial lighting, and 47 % chewed their cage. For 60 % of the ferrets, body weight was higher in winter and lower in summer. Neutered ferrets (58·5 % of males and 62·9 % of females) appeared lighter than intact ferrets of comparable age. A majority (52·4 %) of ferrets received a mix of commercial foods and fresh foods, but 28·6 % were offered no commercial foods. Data were analysed using several multivariable logistic regression models including age, sex, castration, food type and artificial lighting developed for four clinical outcome (lethargy and/or insulinoma, dental problems, diarrhoea and/or bird-seed stools and alopecia). Predictors of four clinical outcomes (lethargy, dental disease, diarrhoea and alopecia) were examined using multivariable logistic regression, with age, sex, neuter status, food type and artificial lighting as the exposure variables. Aged ferrets were more likely to have lethargy, insulinoma, dental problems and alopecia. Ferrets with artificial lighting were more likely to show alopecia. Additionally, ferrets fed commercial food only or a mixed diet (both commercial food and fresh food) were more likely to have lethargy, insulinoma, dental problems, diarrhoea and/or bird-seed stools compared with ferrets fed fresh food only. We also found a significant association between neutering and alopecia. It is to our knowledge the first description of the French population of the ferret as a companion animal.

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.

Herd-level risk factors for bovine tuberculosis in French cattle herds.

Although officially free of bovine tuberculosis (bTB), France has been experiencing a slight increase in the incidence and geographical spread of the infection. Eradication of bTB requires determining the infection risk factors. Although several studies identifying bTB risk factors have been conducted in the United Kingdom and Spain, no information is currently available regarding bTB risk factors in French cattle. The objective of this work was thus to study the factors associated with the risk of bTB in cattle herds in three French administrative divisions (départements of Ardennes, Côte d'Or and Dordogne). A case-control study was conducted to compare herds having experienced a bTB outbreak between 2012 and early 2014 with randomly selected control herds of the three study départements. A questionnaire of farming practices, inter-herd contacts (e.g. at pasture or via vehicles or materials), and the presence of other domestic species was carried out in the selected herds. Data on other variables of interest included animal movements between farms and potential contacts between cattle and wildlife (e.g. badger and wild boar abundances) were also collected. Multivariable logistic regression and multimodel inference methods were used to assess risk factors related to bTB. A total of 216 herds (72 cases and 144 controls) were analyzed. The two main risk factors were the presence of a recent neighboring outbreak, being defined as a neighboring herd at pasture reported as infected in the past two years (odds ratio (OR)=3.6; population attributable fraction (PAF)=30.7%) and the presence of a farm building for cattle housing or for feed storage located at more than 300-m from inhabited areas (OR=2.3; PAF=27.6%). Another risk factor was related to sharing water points at pasture with a recent neighboring outbreak. Results illustrated the multifactorial nature of bTB dynamics. The risk factors related to recently infected neighboring herds could be attributable to between-herd contacts at pasture and/or to exposure to a common source of infection (environment or wildlife). Moreover, the use of remote farm buildings by wildlife may also play a role in the bTB spread in the French départements studied. The identification of the main risk factors help better understand bTB dynamics and are useful for implementing appropriate and targeted surveillance, biosecurity and control measures in France.

Infection of Siberian chipmunks (Tamias sibiricus barberi) with Borrelia sp. reveals a low reservoir competence under experimental conditions.

Reservoir competence is a key parameter in understanding the role of host species in the epidemiology of multi-host-especially vector-borne-pathogens. With this aim in view, we studied the reservoir competence of the Siberian chipmunk (Tamias sibiricus barberi) recently introduced into Europe, for the multi-host tick-borne bacteria, Borrelia burgdorferi sl, the agent of Lyme borreliosis. T. sibiricus were experimentally exposed to bites from Ixodes ricinus ticks infected with Borrelia burgdorferi sensu stricto and Borrelia afzelii, with subsequent assessment of bacteremia and antibody responses. Borrelia was detected in chipmunk blood samples, ear biopsies and organ necropsies, and in nymphs used for xenodiagnosis (at one and six months after the initial chipmunk infection) via both serological and molecular methods. In total, eight out of twelve chipmunks showed evidence of infection by Borrelia sp., either by ELISA or PCR. Five chipmunks developed an immune response against the bacteria one month after infection. Borrelia infection in at least one organ was observed in seven animals at 14, 38, 93 or 178 days post-infection. Xenodiagnosis was positive for one chipmunk at 38 days, but no longer at 178 days post-infection. Four chipmunks remained uninfected, despite similar infection pressures to those observed in the field. Taken together, these results suggest that chipmunks can be infected through Borrelia-infected tick bites, and can transmit Borrelia to nymphs, but do not remain persistently infected.

High-throughput sequence typing reveals genetic differentiation and host specialization among populations of the Borrelia burgdorferi species complex that infect rodents.

Lyme disease is a zoonosis caused by various species belonging to the Borrelia burgdorferi bacterial species complex. These pathogens are transmitted by ticks and infect multiple, taxonomically distinct, host species. From an epidemiological perspective, it is important to determine whether genetic variants within the species complex are able to spread freely through the whole host community or, instead, if certain variants are restricted to particular hosts. To this end, we characterized the genotypes of members of the B. burgdorferi species complex; the bacteria were isolated from more than two hundred individuals captured in the wild and belonging to three different rodent host species. For each individual, we used a high-throughput approach to amplify and sequence rplB, a housekeeping gene, and ospC, which is involved in infection. This approach allowed us to evaluate the genetic diversity both within and among species in the B. burgdorferi species complex. Strong evidence of genetic differentiation among host species was revealed by both genes, even though they are, a priori, not constrained by the same selective pressures. These data are discussed in the context of the advancements made possible by multi-locus high-throughput sequencing and current knowledge of Lyme disease epidemiology.