Leishmania Infection in Wild Lagomorphs and Domestic Dogs in North-East Spain.

Leishmania infantum is a zoonotic protozoan parasite distributed worldwide that is transmitted by phlebotomine sandflies. Dogs are the main reservoir for human infections. However, in recent years, the capacity of lagomorphs to contribute to Leishmania transmission has been confirmed. The present study aimed to assess Leishmania spp. exposure and infection in lagomorphs and sympatric domestic dogs in NE Spain. Sera from European hares, European rabbits, and rural dogs were tested for antibodies against L. infantum using an in-house indirect ELISA. PCR analysis targeting Leishmania spp. was performed in spleens from L. europaeus. Antibodies against Leishmania spp. were detected in all the species analyzed. Total sample prevalence was significantly higher in O. cuniculus (27.9%) than in L. europaeus (2.0%). Results of the PCR were all negative. The present study expands knowledge about Leishmania infections in free-ranging lagomorphs in the Iberian Peninsula, suggesting a more important role of O. cuniculus in the study area. Given the strong correlation between lagomorph densities and human leishmaniasis outbreaks in Spain, the high rabbit and human densities in NE Spain, and the high Leishmania spp. seroprevalence in rabbits, it becomes imperative to establish surveillance programs for lagomorphs in this region.

Pathomorphological Findings and Infectious Diseases in Selected European Brown Hare (Lepus europaeus Pallas, 1778) Populations from Schleswig-Holstein, Germany.

In the northernmost German federal state Schleswig-Holstein, populations of European brown hares (Lepus europaeus Pallas, 1778) show diverse densities and varying courses over the years. To examine differences in pathomorphological findings and infectious diseases as possible reasons for varying population dynamics, we assessed 155 hunted hares from three locations in Schleswig-Holstein from 2016 to 2020. We investigated the association of location, year, age, and sex of animals to certain pathomorphological findings and infectious diseases. Frequent pathomorphological findings were intestinal parasites (63.9%), hepatitis (55.5%), nephritis (31.0%), steatitis (23.2%), enteritis (13.5%), and pneumonia (5.2%). Body condition differed significantly between locations, and the prevalence of pneumonia was significantly higher in females. Enteritis was not detected in 2019, when much more juveniles were sampled. Hepatitis and nephritis occurred significantly more often in 2016 and among adults. Additionally, more adults showed hepatitis with concurrent serotitre for European brown hare syndrome virus (EBHSV), while intestinal parasitosis as well as high excretion rates of coccidia were more common in juveniles. Sampled animals showed high infection rates with Eimeria spp. (96.1%), Trichostrongylus spp. (52.0%), Graphidium strigosum (41.2%), and a high seroprevalence (90.9%) for EBHSV, without severe symptoms. This study revealed a low prevalence of infectious pathogens, but a high prevalence of chronic inflammations of unknown origin in the tested brown hare populations. Overall, our results indicate a rather minor importance of infectious diseases for observed population dynamics of analysed hare populations in Schleswig-Holstein.

"Till death do us apart": The common destiny of brown hare and its parasite community.

Although parasites may threaten individual hosts' survival and reproduction, their role as an essential part of ecosystem functioning and biodiversity has been recognized. In Northern Italy, the hare population has evidently declined since 2008. This paper aims to assess the relationship between host demographic trends and helminth parasite community diversity in a two-year survey in Northern Italy to evaluate the impact of parasites on hosts and confirm the hypothesis that endangered host populations are poor in parasites. In 2013 and 2015 the viscera of 54 and 61 hares legally hunted in agro-ecosystems of the Po Plain were collected. This area is characterized by heavy anthropic pressure: more than 60% of the landscape is represented by agricultural or urbanized territories. No intestinal cestodes were detected. Trichuris sp. and Micipsella numidica were collected in 2015 only; Trichostrongylus retortaeformis, Taenia pisiformis cysticercosis and bronchopulmonary lesions caused by small strongyles were observed in both years. The richness and evenness appeared increased in the second year of the survey, although lower than those obtained by literature data from similar populations examined in the last two decades of 1900. The dominant helminth, T. retortaeformis, was more abundant in individuals with higher weight, while pathological findings were mostly unrelated to this nematode infection; this is consistent with a reduced action, even no harm, of the parasites on the individual host or population level. Our results suggest that the conservation of hosts, environment, and parasites cannot be achieved separately and that parasites and wildlife hosts' destinies are intimately linked, confirming the complexity of ecosystems and the need to contemplate parasite biodiversity in conservation strategies.

Population Density of European Hare Predicts Risk of Tularemia Infection, Czech Republic, 2006-2022.

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.

New agri-environmental measures have a direct effect on wildlife and economy on conventional agricultural land.

The objective of this article is to evaluate economic profits along with return on investment and also the impact of newly designed agri-environmental measures (AEM) on the presence of bioindicator species-European hare and roe deer-in comparison to conventionally cultivated agricultural land. The abundance of European hare was, on average, 4.5-6.7 times higher on AEM compared to the standard agricultural regime and 3.5-6.4 times higher in the case of roe deer in 2020 and 2021. From an economic point of view, the highest incomes were found for extensive orchard alleys and standard conventional crops-wheat and rapeseed rotation. The cash flow from extensive orchard was 4.3 times larger and wheat and rapeseed were 3.5 times larger than from the clover grass mixture. Moreover, the lowest value of operational expenses was found in the case of extensive orchard alleys. The payback period ranged from 16.02 years (wheat and rapeseed rotation) to 53.6 years (clover grass mixture). It is crucial not to assess the economic parameters separately but optimize them with sustainable wildlife management and other benefits that provide ecological and efficient directions of AEM for future generations. However, the performed economic analysis highlights the significantly lower incomes of most components of AEM. We see a principal issue of AEMs usage in the lack of strong incentives for farmers to maximize conservation outcomes. Therefore, the AEMs are often placed in locations with presumed low agriculture profit, which is often related to insignificant conservation effects. Thus, the incomparable AEMs profitability compared to conventional agriculture has to be reflected by the agricultural policy at the European Union level and subsidy policy of particular member states.

Parasitological Survey in European Brown Hare (Lepus europaeus Pallas, 1778) Breeding Facilities in Southern Italy.

Parasites are considered important regulating factors of hosts' population dynamics, not only in free-ranging wildlife, but also in captive bred animals. To date, only few studies have been carried out to assess the parasitic communities of the European brown hare in Southern Italy, and only one focused on animals in captivity. The aim of the present survey was to assess the composition of the endoparasite community in game hares bred for restocking purposes. For this purpose, 215 fecal pools collected in eight different breeding facilities were examined by qualitative and quantitative coprological techniques. Parasites characterized by a direct life cycle, including six species of coccidia from the genus Eimeria and the nematode Trichostrongylus retorataeformis, proved to be the most prevalent parasites. Further helminthic infestations by Passalurus ambiguous, Strongylosides papillosus, Cittotenia spp. and Dicrocoelium dendriticum were also detected, but with an overall prevalence lower than 20%. The present study contributes to increasing knowledge on the health status of a poorly investigated species, and is useful for optimizing breeding efforts in captivity.

Status of Infectious Diseases in Free-Ranging European Brown Hares (Lepus europaeus) Found Dead between 2017 and 2020 in Schleswig-Holstein, Germany.

The European brown hare (Lepus europaeus) is a quite adaptable species, but populations have been decreasing for several decades in different countries, including Germany. To investigate infectious diseases as possible influences on observed population decline in the German federal state Schleswig-Holstein, 118 deceased free-ranging European brown hares were collected between 2017 and 2020 and underwent detailed postmortem examination with extended sampling. Infectious diseases were a major cause of death (34.7%). The number of juveniles found exceeded the adult ones. The main pathomorphological findings were hepatitis (32.8%), pneumonia (22.2%), nephritis (19.1%), liver necrosis (12.9%), and enteritis (40.7%). An unusual main finding was steatitis (20.9%) of unknown origin. Animals were mainly emaciated and showed high infection rates with Eimeria spp. (91.3%) and Trichostrongylus spp. (36.2%). European Brown Hare Syndrome Virus reached an epidemic status with few fatal infections (4.2%) and high seroprevalence (64.9%), whereas the prevalence of Rabbit Haemorrhagic Disease Virus 2 was very low (0.8%) in hares in Schleswig-Holstein. Pathogens such as Yersinia pseudotuberculosis (5.9%), Pasteurella multocida (0.8%), and Staphylococcus aureus (3.4%) only caused sporadic deaths. This study illustrates the wide distribution of various infectious pathogens with high mortality and even zoonotic potential. Infectious diseases need to be considered as an important influence on population dynamics in Schleswig-Holstein.

European Hares, Lepus europaeus, Represent a Reservoir Host for Thelazia callipaeda in Romania.

Thelaziosis caused by Thelazia callipaeda is an emerging disease in Europe. Only two reports of naturally infected lagomorphs have been published so far. The aim of this study was to evaluate the status of the Romanian populations of European brown hares, Lepus europaeus as reservoir hosts for T. callipaeda. Between November 2019 and November 2021, the eyes of 326 L. europaeus carcasses were examined for the presence of ocular parasites. Nematodes were stored in plastic vials with physiological saline, followed by morphological and molecular identification. QGis 3.20 and EpiInfoTM 7 were used for mapping and statistical analysis. Four (1.23%) hares harbored T. callipaeda infection, with a total of 84 nematodes collected (mean intensity 21 nematodes/host), with 45 males, 39 females (two sexually immature, seven with only eggs, and 30 with eggs and larvae). One specimen from each host was successfully sequenced resulting in a 100% similarity with several other sequences of T. callipaeda haplotype 1. Statistical analysis revealed no significant results. The current study represents a first report of T. callipaeda in the European brown hare in Romania, and the second in Europe, also reiterating the role of lagomorphs as reservoir hosts for this zoonotic ocular nematode.

Absence of anti-Besnoitia spp. specific antibodies in European wild lagomorphs from the Iberian Peninsula.

Besnoitia besnoiti is an apicomplexan parasite whose life cycle is not completely understood. It is assumed that this parasite might have an indirect life cycle with a carnivore as a definitive host able to shed oocysts after the ingestion of mature cysts in tissues of an infected intermediate host. Cattle and wild cervids on the Iberian Peninsula can act as intermediate hosts of B. besnoiti, and exposure to the parasite has been demonstrated in equids. In this study, we aimed to assess the presence of members of the genera Besnoitia in wild lagomorphs from the Iberian Peninsula and the potential role of these host species in the life cycle of B. besnoiti, as all the animals were sampled from 19 regions of the Iberian Peninsula where cases of bovine besnoitiosis have been previously detected. Serum samples (Oryctolagus cuniculus: n = 552; Lepus europaeus: n = 122) were first analysed by ELISA and subsequently confirmed by Western blot (WB). Specific antibodies against B. besnoiti were not found in any sampled animal by WB. In addition, lung samples from a subset of wild rabbits (n = 16) were tested by PCR and Besnoitia spp. DNA was not detected. These results suggest that Besnoitia spp. are unlikely to circulate in wild lagomorphs in the Iberian Peninsula. Thus, lagomorphs are not expected to play a key role in the biological cycle of B. besnoiti. Further studies are necessary to assess whether different micromammal species, such as rodents, can serve as natural reservoirs of Besnoitia spp. in other European regions.

Helminth biocoenosis of Lepus europaeus meridiei (Hilzheimer, 1906) from Pianosa island, Italy.

Pianosa is a 10 km2 Italian island in the Tyrrhenian Sea which is part of the Tuscan Archipelago National Park. In this island lives a brown hare population which, according to the literature, belongs to the ancestral taxon Lepus europaeus meridei that offers a unique opportunity to observe how the parasite biocoenosis shapes in condition of isolation, limited space availability and high population density. The aim of this work is to describe the helminth component community of a non-managed, isolated, and dense hare population, evaluating host-parasite relationship and parasite community structure. All 26 analyzed hares (13 males and 13 females) were in good physical conditions, and all of them harboured exclusively the nematode Protostrongylus oryctolagi only. This is the first report of this lungworm species in Italy. The estimated overall abundance was 48.15 worms per examined hare (range 3-258, median 50) and the parasites were unevenly distributed across host population, with few hosts having most parasites (aggregated or overdispersed distribution). No significant relationship was detected between the number of isolated parasites and hare sex and weigh. The effect of the isolation of Pianosa's hare population seems to have acted reducing parasite richness, while the high host density is probably the cause of the high prevalence and abundance of the single helminth species collected. In conclusion, despite the low impact of parasites confirmed also by the overdispersed parasite distribution, the low diversity of the studied parasite community sounds a warning for the management of the hare population and the whole Pianosa's ecosystem.

Logistic Regression Model for Determination of the Age of Brown Hare (Lepus europaeus Pall.) Based on Body Weight.

We developed an algorithm to classify brown hares into two age classes, juveniles (up to 1 year old) and adults (over 1 year old), based on body weight, which can be determined by both the examination of live animals and postmortem analysis. Considering the strong correlation between lens weight and carcass weight, we assumed that hares could be classified into one of the two age groups based only on carcass weight, using a logistic regression model. Using logistic regression, a model was constructed to assess the age of hares based on their body weight. For comparison with the current age-assessment method based on the dry lens weight, a logistic regression classifying the hares based on the dry lens weight was performed as well. The results of the study facilitated the development of a method to classify hares into age groups based on body weight. The proposed approach is innovative, as it allows for the determination of the age of not only culled (postmortem) but also live hares. The method is easy and does not require laboratory tests; hence, the results can be used immediately following evaluation. This method allows hares to be categorized into two age groups (juveniles and adults). With an accuracy of 97.52% and 95.45% in the case of juvenile and adult hares, respectively, the proposed approach can be widely used both in population management and scientific research.

Detection of Anaplasma phagocytophilum in European brown hares (Lepus europaeus) using three different methods.

European brown hare (Lepus europaeus Pallas 1778) is a broadly distributed lagomorph species in Europe, recognized as a host for Ixodes ricinus and reservoir of a wide range of pathogens with zoonotic potential. Even though Lepus europaeus represents an important game animal in Central Europe, the data available on Anaplasma phagocytophilum in this lagomorph are scarce. In this study, three populations of brown hare from distinct localities in the Czech Republic were analysed for the presence of Anaplasma phagocytophilum DNA. We used standard qPCR, targeting the msp2 gene and adapted the same assay also for digital droplet PCR. Out of 91 samples, these two methods identified 9 and 12 as positive, respectively. For taxonomic analysis, we amplified the groEL gene from five of six samples that were found positive by both methods. In phylogenetic analyses, this haplotype belongs to ecotype 1, and to the subclade with isolates from cervids and I. ricinus. Our findings underline the importance of correct result interpretation and positivity cut-off set-up for different detection methods of A. phagocytophilum. This bacterium is characterized by a high intraspecific variability and highly sensitive detection itself, is not enough. Detailed molecular typing is necessary to define the zoonotic potential of different strains and their natural reservoirs.

The influence of preferred habitat and daily range of the European hare on its contamination by heavy metals: a case study from the West Carpathians.

The Spisská Magura mountain range, located in the Middle Spis, is one of the regions in Slovakia most contaminated by heavy metals resulting from mining and smelting activities. Heavy metals and other potentially toxic elements have accumulated in mountain areas via atmospheric transport. The influence of the daily range size of the European hare on its contamination by heavy metals was investigated in three habitat types (forest, woodland edge, meadow) in the Spisská Magura mountain range in the West Carpathians. Individual hares (n = 21) were traced and located by GPS following snowfall. Pair samples of their faeces (n = 64) and food (n = 64) were collected from feeding sites. The maps created were used for determination of the size of the daily range as being small or large. All hares that have a small daily range avoid meadows and open spaces due to the higher predation risk. However, individuals with a large daily range feed in all habitats, including meadows. Hares with a small daily range in a forest habitat ingested higher amounts of bio-elements Ca, Cr, S, and Mn as well as higher amounts of heavy metals Ba and Pb than hares with a large daily range. Moreover, dominant hares with a small daily range, with access to abundant food sources in a forest habitat, may gradually take on higher levels of bio-elements including heavy metals that are present in their food source. In contrast, in the woodland edge, hares with a small daily range had a smaller concentration of Ca, Cr, Mn, S, Ba, and Pb compared to hares with a large daily range. Caecotrophy plays a very significant role as far as the intake of nutrients and other elements is concerned. We found significant dependence between concentrations of the elements Cr, S, Ba, Pb, and Cd in the food of European hares and in their faeces.

Spatiotemporal Overlap between the European Brown Hare and Its Potential Predators and Competitors.

Analysis of spatiotemporal partitioning is pivotal to shed light on interspecific coexistence. Most research effort has involved large-sized carnivores and their prey, whereas little attention has been devoted to lagomorphs. We assessed spatiotemporal overlap among the European brown hare Lepus europaeus and its potential competitors and predators through camera-trapping in an area in Central Italy. We estimated the interspecific patterns of the spatiotemporal activity rhythms of brown hares, its potential predators (the red fox Vulpes vulpes, the pine marten Martes martes, the domestic cat Felis catus, and the domestic dog Canis familiaris), and a competitor, the roe deer Capreolus capreolus. Brown hare activity was studied in natural conditions as well as in a fenced area that excluded terrestrial predators and competitors. Free-ranging hares developed a more nocturnal behavior to avoid diurnal predators (i.e., domestic carnivores and martens). Although high temporal overlap was observed between free-ranging brown hares and both red foxes (82%) and roe deer (81%), hares avoided fox by being more active on darkest nights, as well as avoided roe deer through spatial partitioning. We suggest that hares may adapt their spatiotemporal behavior to avoid potential predators and competitors.

Exploring the Gut Microbiome Alteration of the European Hare (Lepus europaeus) after Short-Term Diet Modifications.

This study aimed to characterise the gut microbiome composition of European hares (Lepus europaeus) and its potential changes after a short-term diet modification. The high sensitivity of European hare to habitat changes makes this species a good model to analyse possible alterations in gut microbiome after the introduction of additional nourishment into the diet. In total, 20 pairs were chosen for the experiments; 10 pairs formed the control group and were fed with standard fodder. The other 10 pairs represented the experimental group, whose diet was integrated with apples and carrots. The DNA from fresh faecal pellets collected after 4 days from the start of the experiment was extracted and the V3-V4 hypervariable regions were amplified and sequenced using the Illumina MiSeq® platform. The obtained amplicon sequence variants were classified into 735 bacterial genera belonging to 285 families and 36 phyla. The control and the experimental groups appeared to have a homogenous dispersion for the two taxonomic levels analysed with the most abundant phyla represented by Bacteroidetes and Firmicutes. No difference between control and experimental samples was detected, suggesting that the short-term variation in food availability did not alter the hares' gut microbiome. Further research is needed to estimate significant time threshold.

Seed traits matter-Endozoochoric dispersal through a pervasive mobile linker.

Although many plants are dispersed by wind and seeds can travel long distances across unsuitable matrix areas, a large proportion relies on co-evolved zoochorous seed dispersal to connect populations in isolated habitat islands. Particularly in agricultural landscapes, where remaining habitat patches are often very small and highly isolated, mobile linkers as zoochorous seed dispersers are critical for the population dynamics of numerous plant species. However, knowledge about the quali- or quantification of such mobile link processes, especially in agricultural landscapes, is still limited. In a controlled feeding experiment, we recorded the seed intake and germination success after complete digestion by the European brown hare (Lepus europaeus) and explored its mobile link potential as an endozoochoric seed disperser. Utilizing a suite of common, rare, and potentially invasive plant species, we disentangled the effects of seed morphological traits on germination success while controlling for phylogenetic relatedness. Further, we measured the landscape connectivity via hares in two contrasting agricultural landscapes (simple: few natural and semi-natural structures, large fields; complex: high amount of natural and semi-natural structures, small fields) using GPS-based movement data. With 34,710 seeds of 44 plant species fed, one of 200 seeds (0.51%) with seedlings of 33 species germinated from feces. Germination after complete digestion was positively related to denser seeds with comparatively small surface area and a relatively slender and elongated shape, suggesting that, for hares, the most critical seed characteristics for successful endozoochorous seed dispersal minimize exposure of the seed to the stomach and the associated digestive system. Furthermore, we could show that a hare's retention time is long enough to interconnect different habitats, especially grasslands and fields. Thus, besides other seed dispersal mechanisms, this most likely allows hares to act as effective mobile linkers contributing to ecosystem stability in times of agricultural intensification, not only in complex but also in simple landscapes.

Genetic diversity and evolution of Hare Calicivirus (HaCV), a recently identified lagovirus from Lepus europaeus.

First recognized as highly pathogenic viruses, hare lagoviruses belonging to genotype GII.1 (EBHSV) infect various Lepus species. Genetically distinct benign lagoviruses (Hare Calicivirus, HaCV) have recently been identified but few data have been available so far on these strains. The analysis of 199 samples from hunted hares collected throughout France allowed the detection of 20 HaCV and showed that they were widely distributed in this country. Ten HaCV capsid protein gene sequences were characterized. A first HaCV capsid protein structural model was proposed, revealing a global structure similar to that of a pathogenic GII.1 strain. The HaCV sequences showed an even higher genetic diversity than previously appreciated, with the characterization of two genotypes (GII.2, GII.3) and several additional putative genotypes. The most recent common ancestor for HaCV VP60 gene was estimated to be much older than that for GII.1 pathogenic strains. These results give new insights into the phylogenetic relationships of HaCV within the Lagovirus genus.

High syphilis seropositivity in European brown hares (Lepus europaeus), Lower Saxony, Germany.

The lagomorph-infecting Treponema paraluisleporidarum is a close relative of the human syphilis-bacterium Treponema pallidum. There is a paucity of information on the epidemiology of hare syphilis and its relationship to the rabbit- and human-infecting treponemes that cause syphilis. In our study, we tested 734 serum samples from European brown hares (Lepus europaeus) collected between 2007 and 2019 in the federal state of Lower Saxony, Germany, for the presence of antibodies against T. paraluisleporidarum. Since T. paraluisleporidarum cross-reacts with T. pallidum antigen, we used a commercially available T. pallidum-particle agglutination (TP-PA) assay to test for the presence of antibodies. A high seropositivity (n = 405/734) was detected. An additional 233 serum samples were retested using a fluorescent treponemal antibody absorption test to confirm the results of the TP-PA assay. Our results show that infection is widespread in Lower Saxony and suggest a horizontal (sexual) transmission mode since adult hares show significantly higher seropositivity than subadults (odds ratio: 0.03 [95% CI 0.02-0.05], p < .0001). No difference was detected based on gender (odds ratio: 0.79 [95% Cl 0.58-1.07], p = .1283). Further studies are warranted to genetically characterize the T. paraluisleporidarum strains that infect wild hares.

European Brown hare (Lepus europaeus) as a source of emerging and re-emerging pathogens of Public Health importance: A review.

European brown hare (Lepus europaeus, EBH) is probably the most important game animal in Europe throughout its historical distribution. The decline in its populations across its geographic range in Europe have been attributed to factors such as reproductive rate and the ability for adaptation, climate, feed availability, predators, anthropogenic factors and diseases. Apart from common diseases of hares with a high impact on their mortality such as European Brown hare Syndrome, EBH has been involved in the epidemiology of pathogens with zoonotic potential. In this work, the role of EBH as a source of Crimean-Congo haemorrhagic fever virus (CCHFV), Hepatitis E virus (HEV), Yersinia spp., Brucella spp., Francisella tularensis, Toxoplasma gondii and Leishmania infantum is discussed. Hares may significantly contribute to the epidemiology of important emerging zoonotic pathogens through maintenance of high endemicity levels as in the case of CCHFV, as a reservoir of important pathogens such as Yersinia spp., B. suis, F. tularensis and L. infantum and as a potential source of T. gondii for other animals, especially for carnivores but also for humans. However, EBH may also be a host of minor importance as in the case of HEV. The continuous surveillance of hare populations will enable the collection of information on the population health status and the pathogens currently circulating in the area posing risk for wildlife, domestic animals and humans. The possible live animal translocations of infected hares, the fact that this species acts as a host of vectors (fleas, ticks, mosquitoes and sandflies) and the prey of carnivores and omnivores that travel in great distances getting into contact with domestic animals and humans, further highlights the need to be included in surveillance studies. Besides, the hunter-harvested EBH population is an excellent indicator for recent pathogen transmission due to its short lifespan.

European hares do not avoid newly pesticide-sprayed fields: Overspray as unnoticed pathway of pesticide exposure.

Although risk assessments for pesticides involve quantifying exposure routes for animals, little empirical evidence exists on how individuals use areas where pesticides were used. Further, the European Food Safety Authority guidelines for the risk assessment of birds and mammals currently only include direct dietary uptake from exposed plants as important pathway for pesticides, arguing that dermal exposure is generally negligible. Here, we use the European hare (Lepus europaeus) as a model of a farmland specialist to investigate if (1) hares adjust habitat use in response to pesticide spraying, using GPS data, and (2) calculate the pesticide exposure and uptake over different pathways, i.e. foraging uptake versus overspray and subsequent oral grooming, using an agent-based modelling approach. Apart from avoidance of sprayed fields on the spraying day by inactive hares, which was likely caused by the mechanical disturbance rather than the pesticide itself, we found no evidence that hares reduced the use of pesticide-sprayed fields compared to control observations where no pesticides were applied. Our simulation showed that both the proportion of exposed individuals and the pesticide uptake were related to the pathway of exposure (foraging versus overspray/oral grooming), and depended on the age class and the vegetation type. We found that pesticide uptake via overspray/oral grooming might be 7-fold higher compared to foraging and might thus be an important pathway of pesticide exposure in hares. Our findings emphasize that policy makers, specifically the European Food Safety Authority, should incorporate alternative pathways of pesticide exposure for birds and mammals, such as overspray and oral grooming, when conducting environmental risk assessments and take variation in vegetation structure and age-related animal behavior into account, because these factors might affect pesticide exposure. Interactions between pesticide application and vegetation structure may both increase or decrease exposure but can be predicted using a simulation approach.