Lepus Europaeus Research Articles

European brown hare syndrome virus and other lagoviruses of interest.

This focus article was prepared by Paul Duff of the APHA Wildlife Expert Group, with support from David Everest (APHA Pathology Department), Lucy Martindale (APHA Surveillance Intelligence Unit), Alex Barlow (Wildlife Network for Disease Surveillance), Mara Rocchi (Moredun Research Institute) and Antonio Lavazza (Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Italy).

The contingent impact of wind farms on game mammal density demonstrated in a large-scale analysis of hunting bag data in Poland

Wind farms are still developing dynamically worldwide, with promising prospects for further growth. Therefore, the assessment of their impact on animals has been carried out. So far, few studies have been conducted on game mammals, and their results are divergent. Previous studies on the impact of wind farms on game species were typically based on regional research covering one or, at most, several wind farms. In this study, we aimed to verify the effect of wind farms on the density of game mammals through a large-scale analysis at the country level, using lowland Poland as an example. The study was based on hunting bag data from open-field hunting districts. It covered seven game species: roe deer (Capreolus capreolus), wild boar (Sus scrofa), red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), European badger (Meles meles), European polecat (Mustela putorius), and European hare (Lepus europaeus). We used Corine Land Cover to account for differences in land cover and the area covered by wind farms in generalized linear mixed models. The study showed that in agricultural landscapes, mainly herbivorous species of game mammals were related to land cover types. These species tend to exhibit higher densities in agricultural areas containing more natural landscape features. Conversely, mesocarnivores are primarily driven by the abundance of prey with little to no observable effects from land cover types. Only roe deer and wild boar presented lower densities with an increase in the area covered by wind farms (for roe deer: estimate: − 0.05, 95% CI: − 0.1–0.0; for wild boar: estimate: − 0.03, 95% CI: − 0.11–0.05), while no effect was observed for mesocarnivores or European hare. The underlying reasons for these relationships remain unclear and require more specific studies. The uncertainty regarding the cause of the observed effects did not allow for a large-scale assessment of the impact of further wind energy development on the studied game mammals.

Dune Hares: Population Indices, Home Range Size, and Habitat Selection of the European Hare on a Danish Island.

Population indices, such as transect counts of animals, can provide important information concerning population changes over time. Moreover, data concerning the home range size and habitat selection of individuals can provide valuable insight into spatial requirements of animals and how they can adapt to variable environments. Here, we describe the population development of European hares (Lepus europaeus) and investigated home range sizes and habitat selection of six radio-tagged individuals on the small (80 ha) Danish Wadden Sea island Langli. The average minimum hare population density from 1983 to 1997 was 64 ± 36 (mean ± SD) hares/km2, with hare numbers varying among years and seasons. The average home range size was 23.3 (CI: 18.9-28.7) ha, which is comparable to agricultural areas of high structural diversity. Moreover, hare habitat selection was generally bimodal, with hares moving over larger areas and selecting marsh habitat for foraging during nighttime, and dune and grassland habitat for resting during daytime, especially during winter. Combined, our results indicate that hare abundance and space use in the dunal landscape of Langli Island were similar to agricultural areas of comparatively high habitat quality. Thus, dunal marsh landscapes offer high-quality habitat for hares and might be of importance as population strongholds at a time when hare populations are declining in many agricultural areas across Europe.

Diet niches of the Alpine mountain hare (Lepus timidus varronis) and European hare (Lepus europaeus) living in coexistence in the Alps

Diet niches of the Alpine mountain hare (Lepus timidus varronis) and European hare (Lepus europaeus) living in coexistence in the Alps

Adaptation of farmland mammalian specialist to urban life: Escape behavior of European hare along the urban-rural gradient

The European hare Lepus europaeus is an iconic but rapidly declining farmland specialist with recently confirmed populations in urban areas. However, their behavioral responses and adaptability to urbanization and life in human-dominated areas are fully unexplored. Here, using infrared cameras, we explored escape behavior (measured as the flight initiation distance) using 965 hare observations in urban and farmland areas in the Czech Republic and Austria (Central Europe) and its association with habitat type, distance to the city center, patch size, season, hare age and initial behavior. We found that European hares adjusted their escape behavior to habitat type and escaped significantly earlier in farmland (rural) habitats than in urban habitats. However, escape distances of hares did not differ between farmland types with different degree of habitat heterogeneity. We also revealed that urban hares escaped earlier when located further from the city center or in a larger study patch. Moreover, adult hares escaped earlier than subadults and foraging individuals escaped earlier than resting hares but both only in rural areas. Our results support high behavioral adaptability of European hares to human-dominated urban areas.

Estimation of mammalian wildlife density by REM method in a Mediterranean forest ecosystem (Pinus brutia) of Türkiye: how human footprint effects mammal community?

BackgroundDetermining the density of mammalian wildlife in an ecosystem is very important for the ecosystem conservation. The aim of this study is to reveal mammalian wildlife density and compare the effect human footprint index (HFI) on the Mediterranean ecosystems. The mammalian wildlife surveys were conducted between August 2013 and December 2013, using 21 camera traps with 2175 camera trap days in a Mediterranean forest ecosystem in İzmir, Türkiye. We used random encounter model (REM) method to estimate densities.ResultsThe population density for 5 mammals were; for red fox 7.89 ind./km2 (± 0.82 SE), wild boar 4.36 ind./km2 (± 0.46 SE), European hare 15.33 ind./km2 (± 03.37 SE), beech marten 0.99 ind./km2 (± 0.10 SE) and golden jackal 0.50 ind./km2 (± 0.05 SE). These results were compared with mammal community which was previously studied in another Mediterranean ecosystem in Muğla, Türkiye, includes caracal and has a lower human footprint index.ConclusionsAccording to results of this study human activity which can be revealed by Human footprint index (HFI) is one of the main parameter on Mediterranean ecosystem and it is effecting the density and occurence of species in mammal community. Both a higher human footprint index and the absence of caracal might cause higher density of red fox and European hare in İzmir, Türkiye. This study also suggests that caracal might be a serious suppressor on red fox which could be explained by competition. Caracal may also control the European hare in Mediterranean forest ecosystem of Anatolia. Thus, decreasing human footprint index and maintaining caracal suppressor effect are crucial for the conservation of the whole Mediterranean ecosystem.

The hare syphilis agent is related to, but distinct from, the treponeme causing rabbit syphilis.

The treponemes infecting lagomorphs include Treponema paraluisleporidarum ecovar Cuniculus (TPeC) and ecovar Lepus (TPeL), infecting rabbits and hares, respectively. In this study, we described the first complete genome sequence of TPeL, isolate V3603-13, from an infected mountain hare (Lepus timidus) in Sweden. In addition, we determined 99.0% of the genome sequence of isolate V246-08 (also from an infected mountain hare, Sweden) and 31.7% of the genome sequence of isolate Z27 A77/78 (from a European hare, Lepus europeaus, The Netherlands). The TPeL V3603-13 genome had considerable gene synteny with the TPeC Cuniculi A genome and with the human pathogen T. pallidum, which causes syphilis (ssp. pallidum, TPA), yaws (ssp. pertenue, TPE) and endemic syphilis (ssp. endemicum, TEN). Compared to the TPeC Cuniculi A genome, TPeL V3603-13 contained four insertions and 11 deletions longer than three nucleotides (ranging between 6 and2,932 nts). In addition, there were 25 additional indels, from one to three nucleotides long, altogether spanning 36 nts. The number of single nucleotide variants (SNVs) between TPeC Cuniculi A and TPeL V3603-13 were represented by 309 nucleotide differences. Major proteome coding differences between TPeL and TPeC were found in the tpr gene family, and (predicted) genes coding for outer membrane proteins, suggesting that these components are essential for host adaptation in lagomorph syphilis. The phylogeny revealed that the TPeL sample from the European brown hare was more distantly related to TPeC Cuniculi A than V3603-13 and V246-08.

Diet and foraging role of European hare (Lepus europaeus) on two invasive non-native shrubs: Cytisus scoparius and Rosa rubiginosa in Northwestern Patagonia, Argentina

ABSTRACT Browsing by alien vertebrate herbivores can have both positive and negative impacts on plant invasion. We studied the European hare (Lepus europaeus) diet and foraging behavior focusing on interactions with two invasive non-native shrub species, Cytisus scoparius and Rosa rubiginosa. Fecal pellets were used for microhistological analysis of diet and seed identification. Foraging use of both species was evaluated by browsing rankings. The diet consisted of 28 items, predominating native grasses such as Poa spp. and Festuca pallescens. Among the non-native species, the grasses Hordeum spp. leaves and stems of the shrub C. scoparius, and seed fragments of Rumex acetosella, and rosehip stand out. Fecal pellets contained 431 whole seeds from nine plant species, predominating non-native species. In diet analysis, a portion of damaged rosehip seeds was observed; however, a significant number of viable seeds were previously extracted. This suggests that consumption of R. rubiginosa fruits contributes to seed dispersal, facilitating its spread. In contrast, the browsing of L. europaeus on the vegetative parts of C. scoparius juveniles significantly reduced their cover, which could slow their growth or prevent their spread. These results are an important input for management decisions to prevent or delay the spread of these invasive species.

Endoparasites of rabbits and hares.

Nematode, cestode, protozoan, microsporidian, and pentastomid parasites affect domesticated and wild rabbits, hares, and jackrabbits of the genera Brachylagus, Lepus, Oryctolagus, Pentalagus, and Sylvilagus. Some endoparasite infections are of limited or no significance, whereas others have potentially profound consequences. Accurate identification of endoparasites of rabbits, hares, and jackrabbits is an important facet of the work of veterinary pathologists engaged in lagomorph pathology. Here I review endoparasites from the pathologist's perspective, focusing on pathogenesis, lesions, and implications of infection. Stomach nematodes Graphidium strigosum and Obeliscoides cuniculi are infrequently pathogenic but may cause gastritis and gastric mucosal thickening. Nematodes Passalurus ambiguus, Protostrongylus spp., Trichostrongylus spp., and Trichuris spp. are rarely associated with disease. Adult Capillaria hepatica (syn. Calodium hepaticum) nematodes and non-embryonated eggs cause granulomatous hepatitis in wild Oryctolagus cuniculus and Lepus europaeus, resulting in multifocal, off-white, hepatic lesions, which may be misdiagnosed as hepatic eimeriosis. When the rabbit is an intermediate host for carnivore cestodes, the space-occupying effects of Cysticercus pisiformis and Coenurus serialis may have pathologic consequences. Eimeria stiedai is a major cause of white-spotted liver in O. cuniculus, particularly in juveniles. Enteric coccidiosis is a noteworthy cause of unthriftiness in young animals, and frequently manifests as diarrhea with grossly appreciable multifocal off-white intestinal lesions. O. cuniculus is the natural host for the zoonotic microsporidian Encephalitozoon cuniculi. Infection may be acute and focused mainly on the kidneys, or it may follow a chronic disease course, frequently with neurologic lesions. A latent carrier status may also develop.

Competition between sympatric hare species in the Alps is boostered by climate change and hybridisation

Climatic variation along the elevation gradient promotes the natural parapatric occurrence of the European hare (Lepus europaeus) and Alpine mountain hare (Lepus timidus varronis) in the Alps. Recent data indicate a displacement of mountain hares caused by competition with the European hare. Competitive exclusion might take place at a fine spatial scale and hybrids may sharpen competition. Genetic non-invasive sampling (gNIS) demonstrates to be effective to retrieve information from wild animals. However, based on the accuracy of the differing genetic analysis methods, the selection of the method might decisively influence results. To examine habitat preferences of Alpine mountain hares, European hares and their hybrids with particular interest in the influence of the accuracy of the genetic analysis method on the results, we performed gNIS in Grisons (Switzerland) for four years and compared habitat associations of the genotyped samples. We recorded 137 individuals (i.e., 35 hybrids, 49 European hares, 53 Alpine mountain hares). Combined nuclear and mitochondrial DNA analysis including individual identification revealed to be the most accurate indirect method for the study of habitat preferences of hares. Alpine mountain hares had a narrow habitat breadth and used little habitat diversity. Hybrids showed great similarities in their habitat preferences to European hares. Hybrids might increase the competition in favour of European hares and the displacement of Alpine mountain hares, since they show similar patterns of habitat use to European hares. Ongoing climate change potentiate the niche overlap between species, increasing the risk of Alpine hare decline due to hybridisation and displacement.

Welfare assessment of European brown hares (Lepus europaeus) reared in captivity for their subsequent release

Welfare assessment of European brown hares (Lepus europaeus) reared in captivity for their subsequent release

Brucella spp. in Wildlife of the Lombardy Region, Northern Italy.

Surveillance data collected in the period 2017-20 for Brucella spp. in wildlife of the Lombardy Region in northern Italy were used to describe the exposure of the wildlife species to Brucella spp. in wild boar (Sus scrofa), European brown hare (Lepus europaeus), fallow deer (Dama dama), red deer (Cervus elaphus), and roe deer (Capreolus capreolus). Among the tested species, wild boar (n=6,440) showed the highest percentage of seropositive samples (5.9%). Notably, wild boars of perifluvial area of the Po River showed higher percentages of positivity than those of the pre-Alpine district. In addition, during the hunting season in 2018, 95 organs (uterus or testes, spleen, and submandibular lymph nodes) from wild boar of the perifluvial area of the Po River were collected for bacteriological examination. Brucella suis was isolated in culture from 18.9% of tested lymph nodes. These serological and microbiological results highlight the presence of B. suis in wild boar and suggest the importance of wild boar as a reservoir for B. suis. Comparison of the spatial distribution of Brucella-seropositive wild boars with the location of backyard swine farms revealed a higher chance of contact between the two populations only in the areas where the lower percentage of seropositive samples was observed. Conversely, the high percentage of seropositive samples observed in the Po River area coupled with positive microbiological cultures suggest a greater risk of infection for the humans directly or indirectly involved in wild boar hunting activity. These results may serve as a basis to establish sound wildlife management and to adopt education campaigns aimed at reducing the risk of human infection in people involved in wild boar hunting related activities.

Rabbit hemorrhagic disease virus 2, 2010-2023: a review of global detections and affected species.

Rabbit hemorrhagic disease virus 2/genotype GI.2 (RHDV2/GI.2; Caliciviridae, Lagovirus) causes a highly contagious disease with hepatic necrosis and disseminated intravascular coagulation in several Leporidae species. RHDV2 was first detected in European rabbits (Oryctolagus cuniculus) in France in 2010 and has since spread widely. We gather here data on viral detections reported in various countries and affected species, and discuss pathology, genetic differences, and novel diagnostic aspects. RHDV2 has been detected almost globally, with cases reported in Europe, Africa, Oceania, Asia, and North America as of 2023. Since 2020, large scale outbreaks have occurred in the United States and Mexico and, at the same time, cases have been reported for the first time in previously unaffected countries, such as China, Japan, Singapore, and South Africa, among others. Detections have been notified in domestic and wild European rabbits, hares and jackrabbits (Lepus spp.), several species of cottontail and brush rabbits (Sylvilagus spp.), pygmy rabbits (Brachylagus idahoensis), and red rock rabbits (Pronolagus spp.). RHDV2 has also been detected in a few non-lagomorph species. Detection of RHDV2 causing RHD in Sylvilagus spp. and Leporidae species other than those in the genera Oryctolagus and Lepus is very novel. The global spread of this fast-evolving RNA virus into previously unexploited geographic areas increases the likelihood of host range expansion as new species are exposed; animals may also be infected by nonpathogenic caliciviruses that are disseminated by almost all species, and with which genetic recombination may occur.

A complete reversal in the effects of extrinsic factors on the escape behaviour of European hares over 33 years

Abstract The flight initiation distance (FID) of prey when escaping from predators is determined by both intrinsic and extrinsic factors. We examined the long-term variation of FID and the impact of various factors on the FID of the European hare, using an economic model. We showed that the effects of local neighbour density and the occurrence of free-ranging dogs on the FID reversed from positive to negative, and the effect of resting in wild vegetation compared with crops increased from the 1960s to the 1990s, in a population in western Poland. The change in FID related to these factors might be explained by changes in agricultural landscape, hare density, and predation risk. The FID of hares reflects their risk-taking behaviour and might change in response to habitat deterioration and changes in prey and predator populations.

Attraction of microarthropods to dung of invasive mammals in meadows of northwestern Patagonia

AbstractThe attractive effect of dung on coprophagous beetle communities and their role in dung degradation are well‐known phenomena. However, little research has been conducted on the recruitment capacity of this substrate for other arthropods, especially those inhabiting the soil. With the aim of beginning to recognize general patterns in these relationships, the influence of the structure and origin of dung from invasive exotic mammals on the recruitment of microarthropods was investigated in two meadows within Nahuel Huapi National Park in southwestern Argentina. Abundance, richness and diversity of microarthropod communities collected on invasive mammal dung were studied. A total of 2478 microarthropods were collected and 52 species were identified. Recruitment of microarthropods on dung of two invasive coprophagic herbivores, the rabbit, Oryctolagus cuniculus and the hare, Lepus europaeus, were compared with two invasive non‐coprophagic mammals, one herbivorous, the red deer, Cervus elaphus and one omnivorous, the wild boar, Sus scrofa. Pellets of S. scrofa showed the highest microarthropod abundance and richness while diversity was greatest on pellets of C. elaphus. Pygmephorid and oribatid mites and springtails were the most abundant. Liposcelis psocopterans and oribatid mites were more abundant on S. scrofa dung. Low similarity was observed when comparing dung of invasive mammals from different sites revealing spatial variability regarding species composition. Based on its more diverse diet, we conclude that S. scrofa dung exerts a strong influence on microarthropod communities by recruiting a greater number of individuals and species than the dung of the remaining invasive mammals.

Leptospira borgptersenii and Leptospira interrogans identified in wild mammals in Rio Grande do Sul, Brazil.

Leptospira spp. are bacteria responsible for leptospirosis, a zoonotic disease with considerable impacts on the economy, animal health, and public health. This disease has a global distribution and is particularly prevalent in Brazil. Both rural and urban environments are habitats for Leptospira spp., which are primarily transmitted through contact with the urine of infected animals. Consequently, domestic and wild species can harbor these prokaryotes and serve as infection sources for other hosts. In the context of wild animals, there is a dearth of molecular studies elucidating the roles of various animal and bacterial species in the epidemiology of leptospirosis. Therefore, this study aimed to evaluate the presence of Leptospira spp. DNA in different species of free-living and captive wild animals and to assess the phylogenetic relationships of the identified microorganisms in Rio Grande do Sul, Brazil. The samples were evaluated for the presence of the gene lipL32 by polymerase chain reaction (PCR) and sequencing of the amplified fragment after which phylogenetic analyzes were carried out. DNA from Leptospira spp. was extracted from kidney tissue from wild animals (Mammalia class). Pathogenic Leptospira spp. DNA was detected in 9.6% (11/114) of the samples, originating from nine species of wild animals, including the white-eared opossum (Didelphis albiventris), skunk (Conepatus chinga), geoffroy's cat (Leopardus geoffroyi), margay (Leopardus wiedii), pampas fox (Lycalopex gymnocercus), capybara (Hydrochoerus hydrochaeris), common marmoset (Callithrix jacchus), neotropical river otter (Lontra longicaudis), and european hare (Lepus europaeus). Phylogenetic analysis revealed the presence of Leptospira borgpetersenii and Leptospira interrogans in these animals. This research is the first study contributing to the epidemiology of leptospirosis by identifying L. borgpetersenii and L. interrogans in free-living and captive wild animals in Rio Grande do Sul, Brazil, potentially acting as bacterial reservoirs. Additionally, our findings can inform sanitary measures for controlling and preventing the disease, thereby safeguarding public health.

Toxoplasma gondii infection in European brown hares (Lepus europaeus) in Brazil.

Toxoplasmosis is a worldwide zoonosis that affects warm-blooded animals, including humans. Wild animals can act as intermediate hosts of this pathogen; thus, this study aims to detect Toxoplasma gondii infection in invasive European brown hares in Brazil. For this, 72 wild European brown hares were captured from July 2020 to June 2022 in three Brazilian states: São Paulo, Paraná, and Rio Grande do Sul. The diagnostic of Toxoplasma gondii infection was performed by bioassay in mouse, histopathology in Hematoxylin-Eosin-stained tissue sections (brain, liver, lungs, kidneys, and small intestine), serology by IFAT, and molecular techniques by conventional PCR and qPCR. The combined prevalence of the different diagnostic methods was 51.4% (37/72, CI= 40.1 - 62.6 %), and there was no statistical difference between sexes, age range, or geographical region of the hosts. Mouse bioassay was the technique that detected more positive hares. To our knowledge, this is the first confirmation of Toxoplasma gondii infection in invasive European brown hares in Brazil. These animals act as reservoirs and potential infection source for carnivores and other wild and domestic animals, including humans, thus contributing to perpetuate the disease cycle in São Paulo, Paraná, and Rio Grande do Sul States. Research such as the present study is necessary to raise awareness about the role of animals in the disease cycle.

The Taurus Mountains, the Hotspot of Western Palearctic Biodiversity, Is in Danger: Marble Quarries Affect Wildlife

The Taurus Mountains in the Mediterranean Coastal Basin, considered a biodiversity hotspot, have a rich biodiversity in the Western Palearctic. The number of marble quarries in the Taurus Mountains has dramatically expanded over the past ten years. The objectives of this study are to (i) determine the impacts of quarrying on wildlife and (ii) determine the potential impacts of quarrying on the future of Taurus. A total of 57,547 photos and video images were analyzed on 5447 photo-trap days in two areas, the marble quarries and the control areas. Using 97 randomly selected marble quarries, the area they cover and their annual growth rates were determined. The most commonly seen animals were the wolf (Canis lupus), fox (Vulpes vulpes), lynx (Lynx lynx), and wild boar (Sus scrofa) in the control area, and the jackal (Canis aureus) and hare (Lepus europaeus) in the marble quarries (p < 0.001). Additionally, we found a significant positive correlation between the distance from the geographical center of the marble quarries and the number of dates of wolf, fox and wild boar sightings, with a significant negative correlation for hares (p < 0.05). A positive correlation was found between the area of marble quarries and the duration of operation (R = 0.89, p < 0.00). The waste from quarries, which makes up 79.7% of the total land used for this purpose, is the greatest cause of habitat degradation. According to calculations, even if no new marble quarries are built as of right now, 7.14% of the Taurus Mountains may have disappeared by the year 2027, and by the year 2032, 8.25% of the Taurus ecosystems may have disappeared completely. The Taurus Mountains, a center of Western Palearctic biodiversity, are being threatened by marble quarries. This study advances our knowledge of how marble quarries may affect wildlife. New strategies must be developed as soon as possible to protect the Taurus Mountains, the hotspot of the Mediterranean basin.

The low survival rate of European hare leverets in arable farmland: evidence from the predation experiment.

The low survival rate of leverets may significantly contribute to steep population declines and slow recovery of European hares (Lepus europaeus). However, the leveret survival rate in farmlands with different landscape structures is poorly understood, and the existing evidence comes mainly from Western Europe. In this study, we explored the survival of leveret hare dummies along linear semi-natural habitats in homogeneous Central European arable farmland during the main part of the European hare reproduction period (March-April) in 2019 and 2020. The survival rate of hare leverets during the 14-day period was only 22.2%, and all predation events were recorded during the first six days of the experiment. Mammalian predators were responsible for 53.1% of predation events, avian predators for 40.8%, and agricultural operations for 6.1%. The red fox (Vulpes vulpes) was the dominant predator in our study area and was the primary cause of leveret dummy mortality (32.7%), but it also had the highest use-intensity and visit frequency of all of the study plots. Predation by avian predators was associated with patches of lower vegetation height and cover (such as plowed fields) and during daylight hours, whereas the opposite was true for mammalian predators. We propose that improving the habitat quality of arable landscapes by increasing the proportion and quality of extensively used non-farmed habitats (e.g., set-asides, wildflower areas, extensive meadows, fallow land, and semi-natural habitats on arable land) providing cover and shelter for leverets could be an effective management measure for reducing predation risk on leverets.

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.