Wild Rabbit Oryctolagus Cuniculus Research Articles

Epidemiological survey and risk factors associated with Sarcoptes scabiei in wild lagomorphs in Spanish Mediterranean ecosystems

Although mortality caused by Sarcoptes scabiei has been reported in European wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis), there is a lack of detailed information regarding the exposure of wild lagomorph species to this parasite. Here, we aimed to determine the seroprevalence and potential risk factors associated with S. scabiei exposure in European wild rabbits and Iberian hares in Mediterranean ecosystems of southern Spain. Between 2018/2019 and 2021/2022 hunting seasons, serum samples from 464 wild rabbits and 132 Iberian hares were collected from 100 hunting grounds in Andalusia (southern Spain). Sera were tested using an in-house indirect ELISA to detect specific anti-S. scabiei antibodies based on the immunodominant protein Ssλ20ΔB3. The overall apparent individual seroprevalence was 15.9% (95/596; 95%CI: 13.0–18.9). Antibodies against S. scabiei were detected in 11.6% (54/464; 95%CI: 8.7–14.5) of the European wild rabbits and 31.1% (41/132; 95%CI: 23.2–39.0) of the Iberian hares. Species (Iberian hare), age (adults) and geographical area (western Andalusia) were identified as risk factors potentially associated with S. scabiei exposure using generalized estimating equation analysis. By applying spatial analysis, two significant cluster of high seropositivity were detected in western and central Andalusia, respectively. The seroprevalence values obtained provide evidence of endemic, widespread and heterogeneous exposure to S. scabiei among wild lagomorph populations in Spanish Mediterranean ecosystems. Our findings underscore the importance of implementing integrated surveillance programs for sarcoptic mange in wild lagomorphs as well as in other sympatric species.

Is the Iberian lynx a hunters’ ally? a case study from a reintroduced population in Portugal

The Iberian lynx (Lynx pardinus) is one of the most endangered felids in the world, and a high proportion of its populations occur in hunting grounds managed for red-legged partridge (Alectoris rufa) and wild rabbit (Oryctolagus cuniculus), both game species of high socio-economic value and the latter a key prey species for the lynx. In this case study from south-eastern Portugal, we evaluated the game management practices, the abundance of partridges and rabbits (expressed as Kilometric Abundance Index), and the occurrence of mesocarnivores in four hunting grounds, two with and two without breeding lynx. In all grounds game management was conducted but in those where lynx occurred, full-time gamekeepers were employed and hunting was the major source of income. The results showed that grounds with breeding lynx had higher KAI values for partridges and rabbits, together with reduced mesocarnivores occurrence (particularly red fox Vulpes vulpes and stone marten Martes foina), compared to those without breeding lynx. Our results confirm that lynx occurrence fits within the management traits of those hunting grounds in Iberia aiming to increase partridges and rabbits for sustainable hunting, as they act as ‘natural’ gamekeepers. This could be considered as a ‘win-win’ situation; successful for lynx’s establishment and small game conservation.

Occurrence of Coxiella burnetii in wild lagomorphs and their ticks in Spanish Mediterranean ecosystems.

Coxiella burnetii, the causative agent of Q fever, is a zoonotic multi-host vector-borne pathogen of major public health importance. Although the European Food Safety Authority has recently made the monitoring of this bacterium in wildlife a priority, the role of wild lagomorphs in the transmission and maintenance of C. burnetii is poorly understood. The aims of this study were to determine the prevalence and risk factors associated with C. burnetii circulation in European wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis) and to assess the presence of this pathogen in ticks that feed on them in Mediterranean ecosystems in Spain, the country with the highest number of reported cases of Q fever in Europe. A total of 574 spleen samples were collected from 453 wild rabbits and 121 Iberian hares, and 513 ticks (processed in 120 pools) between the 2017/2018 and 2021/2022 hunting seasons. C. burnetii DNA was detected in 103 (17.9%; 95% CI: 14.8-21.1) of the 574 wild lagomorphs tested. By species, prevalence was 16.3% (74/453; 95% CI: 12.9-19.7) in the European wild rabbit and 24.0% (29/121; 95% CI: 16.4-31.6) in the Iberian hare. At least one positive lagomorph was found on 47.9% of the 96 hunting estates sampled and in every hunting season since 2018/2019. Two risk factors associated with C. burnetii infection were as follows: outbreak of myxomatosis on the hunting estate in the month prior to sampling and high tick abundance observed by gamekeepers on the hunting estate. C. burnetii DNA was also found in 33 of the 120 (27.5%; 95% CI: 19.5-35.5) tick pools tested. The pathogen was detected in 66.7% (4/6), 29.2% (26/89) and 21.4% (3/14) of Haemaphysalis hispanica, Rhipicephalus pusillus and Hyalomma lusitanicum pools respectively. This study provides new epidemiological data on C. burnetii in European wild rabbits and is the first survey on this zoonotic pathogen performed in Iberian hares. Our results indicate widespread endemic circulation of C. burnetii and highlight the importance of both wild lagomorph species as natural reservoirs of this zoonotic bacterium in Mediterranean ecosystems in southern Spain, which may be of public and animal health concern. The high prevalence and wide diversity of positive tick species suggest the possible role of ticks in the epidemiological cycle of C. burnetii, with the potential risk of transmission to sympatric species, including humans.

Epidemiological surveillance of Leishmania infantum in wild lagomorphs in Spanish Mediterranean ecosystems

Wild lagomorphs play a key epidemiological role as reservoirs of Leishmania infantum, causative agent of the largest outbreak of human leishmaniosis in Europe to date. A large-scale survey study was conducted on wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis) populations in Spanish Mediterranean ecosystems to evaluate the exposure of L. infantum and investigate potential risk factors associated with exposure to this zoonotic parasite. Between 2018 and 2021, a total of 631 wild lagomorphs (471 wild rabbits and 160 Iberian hares) were collected in Andalusia (southern Spain) and tested for antibodies against L. infantum using the indirect fluorescent antibody test (IFAT). Spleen samples from 563 of the wild lagomorphs sampled (441 wild rabbits and 122 Iberian hares) were also evaluated by real-time quantitative PCR (qPCR) for detection of Leishmania kDNA. Exposure to L. infantum (positive by IFAT and/or qPCR) was detected in 56.4 % (356/631; 95 %CI: 52.3–60.3) of the lagomorphs analyzed. Anti-Leishmania antibodies were found in 12.8 % (81/631; 95 %CI: 10.2–15.5) of the animals, and L. infantum kDNA was detected in 59.0 % (332/563; 95 %CI: 54.9–63.0) of the spleen samples tested. Phylogenetic analysis revealed high homology (99.9–100 %) between L. infantum sequences obtained and strains previously isolated from humans in Spain. While apparent seroprevalence was significantly higher in Iberian hares (19.4 %; 95 %CI: 13.3–25.5) compared to wild rabbits (10.6 %; 95 %CI: 7.9–13.4), no significant differences in prevalence were found between wild rabbits (61.0 %; 95 %CI: 56.5–65.6) and Iberian hares (51.6 %; 95 %CI: 42.8–60.5). At least one positive animal was found on 64.8 % (70/108) of the hunting grounds sampled, and a high-risk spatial cluster (P < 0.001) was also identified in central Andalusia. The multivariable analysis identified bioclimatic level (meso-Mediterranean climate) and the presence of goats on hunting grounds as risk factors potentially associated with L. infantum exposure in wild lagomorphs. This study shows high, widespread exposure, but heterogeneous distribution of L. infantum in wild lagomorph populations in Mediterranean ecosystems in southern Spain. The results point to the need to promote integrated surveillance programs for the detection of Leishmania spp. in wild lagomorphs in order to establish effective control measures against human leishmaniosis under a One Health approach.

Targeted and untargeted metabolomic profiles in wild rabbit does (Oryctolagus cuniculus) of different breeding states (pregnant and lactating).

Ecological nutrition aims to unravel the extensive web of nutritional links that drives animals in their interactions with their ecological environments. Nutrition plays a key role in the success of European wild rabbit (Oryctolagus cuniculus) and could be affected by the breeding status of the animals and reflected in the metabolome of this species. As nutritional needs are considerably increased during pregnancy and lactation, the main objective of this work was to determine how the breeding status (pregnant and lactating) of European wild rabbit does affects nutritional requirements and their metabolome (using targeted and untargeted metabolomics), aiming to find a useful biomarker of breeding status and for monitoring nutritional requirements. To address this gap, 60 wild European rabbits were studied. Animals were divided according to their breeding status and only pregnant (n = 18) and lactating (n = 11) rabbit does were used (n = 29 in total). The body weight and length of each animal were analyzed. The relative and absolute chemical composition of the gastric content and whole blood sample were taken, and targeted and untargeted metabolomics were analyzed. As a main result, there were no differences in biometric measurements, gastric content, and targeted metabolomics, except for live weight and nonesterified fatty acids (NEFA), as pregnant animals showed higher live weight (+12%; p = 0.0234) and lower NEFA acid levels (-46%; p = 0.0262) than lactating females. Regarding untargeted metabolomics, a good differentiation of the metabolome of the two breeding groups was confirmed, and it was proven that pregnant animals showed higher plasmatic levels of succinic anhydride (3.48 more times; p = 0.0236), succinic acid (succinate) (3.1 more times; p = 0.0068) and propionic acid (3.98 more times; p = 0.0121) than lactating animals. However, lactating animals showed higher levels of N-[(3a,5b,7b)-7-hydroxy-24-oxo-3-(sulfoxide) cholan-24-yl]-Glycine (cholestadien) (2.4 more times; p < 0.0420), 4-maleyl-acetoacetate (MAA) (3.2 more times; p < 0.0364) and irilone (2.2 more times; p = 0.0451) than pregnant animals, any of these metabolites could be used as a potential biomarker. From these results, it can be concluded that the most notable changes were observed in the metabolome of individuals, with most of the changes observed being due to energy and protein mobilisation.

Exposure to Coxiella burnetii in Wild Lagomorphs in Spanish Mediterranean Ecosystems.

Coxiella burnetii is an important zoonotic pathogen of worldwide distribution that can infect a wide range of wild and domestic species. The European wild rabbit (Oryctolagus cuniculus) can play a role as a reservoir for this bacterium in certain epidemiological scenarios, but, to date, a very limited numbers of large-scale serosurveys have been conducted for this species worldwide. Although exposure in hare species has also been described, C. burnetii in Iberian hare (Lepus granatensis) has never been assessed. Here, we aimed to determine the seroprevalence and risk factors associated with C. burnetii exposure in wild lagomorphs in the Mediterranean ecosystems of southern Spain. Between the 2018/2019 and 2021/2022 hunting seasons, blood samples from 638 wild lagomorphs, including 471 wild rabbits and 167 Iberian hares, were collected from 112 hunting grounds distributed across all eight provinces of Andalusia (southern Spain). The overall apparent individual seroprevalence was 8.9% (57/638; 95% CI: 6.8-11.4). Antibodies against C. burnetii were found in 11.3% (53/471; 95% CI: 8.4-14.1) of the wild rabbits and 2.4% (4/167; 95% CI: 0.1-4.7) of the Iberian hares. Seropositive animals were detected for 16 (14.3%; 95% CI: 7.8-20.8) of the 112 hunting grounds tested and in all the hunting seasons sampled. A generalized estimating equations model showed that the geographical area (western Andalusia) and presence of sheep were risk factors potentially associated with C. burnetii exposure in wild lagomorphs. A statistically significant spatial cluster (p < 0.001) was identified in the south-west of Andalusia. Our results provide evidence of moderate, endemic and heterogeneous circulation of C. burnetii in wild lagomorph populations in Spanish Mediterranean ecosystems. Risk-based strategies for integrative surveillance programs should be implemented in these species to reduce the risk of transmission of the bacterium to sympatric species, including humans.

Low 25-hydroxyvitamin D concentrations in wild rabbits (Oryctolagus cuniculus) in southern Finland

BackgroundDiet and endogenous vitamin D synthesis are possible sources of vitamin D in wild rabbits. Higher 25-hydroxyvitamin D (25(OH)D) concentrations have been reported in rabbits after artificial UVB light exposure than in rabbits without this exposure, suggesting that endogenous vitamin D synthesis occurs in the former group. In Finnish pet rabbits, diet was reported as main source of vitamin D, while outdoor access was not. Finland’s northern location only enables endogenous synthesis from mid-March to mid-October in people with light skin type. Living conditions during winter are challenging for Finnish wild rabbits. This study aimed to measure serum 25(OH)D concentrations and possible natural seasonal variation of vitamin D concentrations in Finnish wild rabbits.ResultsPost-mortem blood samples (n = 78) were collected between 2013 and 2021 from wild rabbits hunted for reduction of the wild rabbit population. Separated sera were stored at − 80 °C until 25(OH)D concentrations were measured by enzyme immunoassay. Data regarding sex were available from 50 rabbits, 29 (58%) of which were females. Mean 25(OH)D concentration was 3.3 (range 0.3–7.1) ng/ml. 25(OH)D concentration was statistically similar between season (autumn, winter, summer), month or year of sample collection, and sex.ConclusionsWild rabbits living in Finland have very low serum 25(OH)D concentrations. This is far below the previously suggested threshold of vitamin D deficiency in rabbits (17 ng/mL) or the mean 25(OH)D concentration reported in Finnish pet rabbits (26.0 ng/mL). Seasonal variation was not observed in 25(OH)D concentrations between winter and summer months. Even though rabbits are crepuscular animals and may spend the mid-day in underground burrows, the very low observed 25(OH)D concentrations raise doubt about whether vitamin D synthesis occurs efficiently in the skin of rabbits and whether the diet of wild rabbits provides adequate amounts of vitamin D. Cutaneous vitamin D synthesis, possible long-term consequences of low 25(OH)D concentrations, and the association of low vitamin D status with other health disorders warrant further investigations in rabbits.

Occurrence of Anaplasma spp. in wild lagomorphs from Southern Spain: Molecular detection of new Anaplasma bovis lineages

Wild lagomorphs can act as reservoirs of several pathogens of public and animal health concern. However, the number of studies assessing the presence of Anaplasma spp. in these species is scarce. The aim of the present study was to molecularly identify Anaplasma spp. in wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis) from Southern Spain and assess their epidemiological role in the maintenance of the bacterium.During 2017–2021, spleen samples of 394 wild rabbits and 145 Iberian hares were collected. Anaplasma DNA was detected using different PCR assays (16S rRNA and groEL) and phylogenetic analyses were carried out by Bayesian approach. The possible influence of lagomorph species, age and sex on the prevalence of Anaplasma spp. was evaluated by a multiple logistic regression model.The 9.4% of the rabbits were positive to Anaplasma bovis, but all the hares were negative. No significant differences were found in Anaplasma spp. prevalence regarding to age or sex. This is the first report of A. bovis in lagomorphs from Europe. The phylogenetic analysis of A. bovis confirms the existence of different clusters suggesting the existence of several lineages. In addition, a high divergence of nucleotide identity was observed within the lineage 4, which could result in the under-detection of some strains when using A. bovis-specific PCR, hindering its detection and characterization. Since this analysis is based on a limited number of nucleotide bases and sequences, more studies are needed for further characterize A. bovis, as well as its relationship with other Anaplasma spp.

High Mortality of Wild European Rabbits during a Natural Outbreak of Rabbit Haemorrhagic Disease GI.2 Revealed by a Capture-Mark-Recapture Study

Rabbit haemorrhagic disease virus (RHDV) GI.2 has caused significant declines in the abundance of wild European rabbits (Oryctolagus cuniculus), contributing to the species being recently classified as “endangered” in its native range. The epidemiology of this virus is still poorly understood despite its relevance for domestic and wild rabbits. During a longitudinal capture-mark-recapture (CMR) study of wild Iberian rabbits, O. c. algirus, in a semiextensive breeding enclosure, an outbreak of RHDV GI.2 took place in January-February 2022, allowing us to estimate key epidemiological parameters of a natural outbreak. From April 2021 to July 2022, 340 rabbits were captured 466 times and individually identified, and some were vaccinated against myxoma virus (MYXV) and/or RHDV GI.2. Sera were collected and tested for IgG specific for MYXV and RHDV GI.2, and data were analyzed using multievent CMR models. During six weeks in January-February 2022, an estimated 81.0% (CI95 77.1–84.3%) of the population died. Intensive aboveground searches could recover 189 carcasses (50.5% of the estimated mortality, CI95 41.8–63.4%), with RHDV GI.2 detected in 6/7 tested. Apparent RHDV GI.2 seroprevalence rose from 15.4% (CI95 8.0–27.5%) in January 2022 to 87.9% (CI95 72.7–95.2%) in February 2022. The apparent mortality of RHDV GI.2-seropositive rabbits during the outbreak was estimated as null, while for seronegative rabbits, it was 76.0% (CI95 53.8–90.3%). Among the seronegative rabbits, mortality was higher in unvaccinated (100%) than in recently vaccinated (60.0 ± 16.6%) and in females (100%) than in males (52.0 ± 17.1%). Infected carcasses in the burrows might explain the medium-term disease persistence in the population following the outbreak. Rabbits with antibodies at the cutoff for seropositivity were fully protected from fatal infection. Females had a higher fatality rate than males, underscoring the impact of RHDV GI.2 on the population dynamics of this endangered species.

Rabbits or Refuse? Landfill Use and Relevance as a Food Source for an Increasing Wintering Population of the Red Kite

Household waste landfills represent a huge source of trophic resources for opportunistic and versatile wildlife species. Among them, the red kite (Milvus milvus) is one of the most endangered in Europe. Several studies have pointed out the importance of landfills as a source of food for this species during the wintering season, but the information on the frequency, seasonal and daily patterns of use, and age of red kites that exploit this food source is still insufficient to understand their actual role in conservation. In this study, we evaluated the patterns of use of household waste by overwintering red kites in southeastern Madrid, central Spain. The results showed the constant presence of relatively low numbers (&lt;30 on average) and a reduced hourly inflow and outflow of red kites throughout the day and during the whole winter period in the studied landfill. A higher proportion of juveniles was found in the landfill than in the overall wintering population. Pellet analysis clearly shows that the diet of red kites is dominated by the wild rabbit (Oryctolagus cuniculus), which is quantitatively very relevant compared to household waste obtained from the landfill. This suggests a relatively low quantitative importance of landfills as foraging grounds for the increasing population of wintering red kites in the study area. Instead, the high regional density of wild rabbits attracts large numbers of red kites that can eventually use landfills as a non-optimal last-resort foraging option, owing to the predictability of household waste, especially for juveniles. The continuous presence of red kites in landfills likely influences an uninformed positive perception about their relevance to the conservation of the wintering population, despite risks there faced, such as collision, electrocution, and intoxication. Future research is needed to assess in depth the influence of wild rabbits in Spain on the habitat use and global population dynamics of red kites.

Monitoring of Hepatitis E Virus in Wild Lagomorphs in Spanish Mediterranean Ecosystems

Hepatitis E virus (HEV) is an emerging zoonotic pathogen in Europe. Even though swine species are considered the main host of the zoonotic HEV-3 genotype, rabbits are recognized as the main reservoir of the divergent HEV-3ra subtype. However, the role of wild lagomorphs in the epidemiology of this virus in Mediterranean ecosystems is under debate. The aims of this study were to assess exposure of HEV in wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis) populations in southern Spain and to determine potential risk factors associated with HEV exposure in these species. Between 2018 and 2021, blood and fecal samples from 370 wild rabbits and 60 Iberian hares were collected. A total of 29 (6.7%; 95%CI: 4.4–9.1) out of 430 sampled animals showed anti-HEV antibodies. By species, the seroprevalences in wild rabbit and Iberian hare were 6.8% (29/370; 95%CI: 4.2–9.3) and 6.7% (4/60; 95%CI: 0.4–13.0), respectively. Seropositive animals were detected on 17 (26.2%; 95%CI: 15.4–36.8) of the 65 sampled hunting estates. The generalized estimating equations model showed that geographical area was a risk factor potentially associated with HEV exposure in wild lagomorphs in the study region. HEV RNA was not detected in any of the 242 (0.0%; 95%CI: 0.0–1.5) fecal samples tested. This is the first large-scale serosurvey performed in wild rabbits in the Iberian Peninsula and in Iberian hares worldwide. Our results provide evidence of low, widespread, and heterogeneous distribution of HEV among wild rabbit and Iberian hare populations in Spanish Mediterranean ecosystems, which indicates a limited role of wild lagomorphs in the maintenance of the virus and a low risk of transmission of HEV to other species, including humans.

The Role of PRLR Gene Polymorphisms in Milk Production in European Wild Rabbit (Oryctolagus cuniculus).

One of the problematic points of rabbit breeding is that the nutritional requirements of the kits are not fully satisfied by the does' milk production from the third week of lactation onwards. The prolactin receptor gene has a significant effect on reproductive processes, and its polymorphisms have been associated with milk production in several species (cattle, goats, sheep, and buffalo). The European wild rabbit (Oryctolagus cuniculus), has a more diverse genetic background compared to domesticated lines. In the course of our study, sequencing of the 1210 bp long segment of the PRLR gene promoter region was accomplished. We detected four point mutations (SNP1-407G > A, SNP2-496G > C, SNP3-926T> and SNP4-973A > C) and one microsatellite at position 574. In our population, the four SNPs were segregated into four genotypes: AACCCCCC, GGGGTTAA, AAGGTTAC, and GGGGTCAC. Our results show that the genotype in the homozygous form is associated with higher milk production (1564.7 ± 444.7 g) compared to the other three genotypes (AACCCCCC 1399.1 ± 326.8 g; GTGACCTT 1403.8 ± 517.1 g; GGGGTCAC 1220.0 ± 666.2 g), and the short microsatellite repeat (167 bp) also coincides with significantly higher milk production (1623.8 ± 525.1 g). These results make the marker-assisted selection possible also for domesticated lines.

Understanding the impact of wild boar on the European wild rabbit and red-legged partridge populations using a diet metabarcoding approach

European wild rabbit (Oryctolagus cuniculus) and red-legged partridge (Alectoris rufa) are main small game species of conservation concern in Mediterranean ecosystems. To date, their presence in wild boar’s (Sus scrofa) diet and factors driving their consumption have been little investigated. A genetic metabarcoding approach was used on 80 wild boar faeces collected from four hunting estates devoted to small game hunting during two different seasons. Abundances of wild boar, rabbits and partridges were first estimated. Results showed DNA of seventeen chordate species. The highest frequency of occurrence (FO) corresponded to mammals and birds, with 77.6 and 22.3%, respectively. DNA of game species was detected in 39/71 (FO = 55.0%) samples, highlighting the presence of European wild rabbit in 27 (FO = 38.0%) and red-legged partridge in eight (FO = 11.3%). Dietary composition varied between seasons and estates, being rabbit the main responsible (explaining 35.26% and 39.45% of differences, respectively). Rabbit FO in the diet was positively related to the abundance of wild boar and rabbit density on the estate. It was greater in autumn and in estates where rabbits were hunted. Regarding red-legged partridge, a significant and positive relationship between its population density and its diet FO was observed, without significant differences between seasons or estates. Overall, our results suggest wild boar as an opportunistic species whose diet is largely determined by the relative availability of different food resources. Its ecological role concerning small game species in Mediterranean agroecosystems seems to be more related to consumption of carrion during the hunting season than to direct predation.

A new longevity record for a wild European rabbit (Oryctolagus cuniculus) from South Australia

A population of wild European rabbits (Oryctolagus cuniculus) was regularly monitored from November 1996 to April 2022, using mark–recapture as part of a longitudinal epidemiological study into two Australian rabbit biocontrol agents, rabbit haemorrhagic disease (RHD) and myxomatosis. A female rabbit, first captured as a subadult on 26 October 2009, was recaptured 16 times before its final capture on 19 February 2019. The longevity of this rabbit, from its calculated birth date to the date of its last capture, was 9.5 years. This is a record lifespan for a wild European rabbit – acknowledging that such longevity might be specific to our site in temperate South Australia. It exceeds our previous reported record from the same site by 2 years.

Iberian wild leporidae as hosts of zoonotic enteroparasites in Mediterranean ecosystems of Southern Spain.

Wild lagomorphs including rabbits and hares can act as natural carriers or reservoirs of bacterial and parasitic zoonotic diseases. However, little is known on the epidemiology and potential public health significance of intestinal eukaryotes in wild leporids. We examined faecal samples from European wild rabbits (Oryctolagus cuniculus, n=438) and Iberian hares (Lepus granatensis, n=111) collected in the Autonomous Region of Andalusia in southern Spain during 2012-2021. We searched for the presence of DNA from the main intestinal protist and microsporidial pathogens of veterinary and public health concerns using molecular methods (PCR followed by Sanger and next-generation sequencing). Giardia duodenalis was the most prevalent species found (27.8%, 153/550; 95% CI: 24.1-31.8), followed by Cryptosporidium spp. (1.3%, 7/550; 95% CI: 0.5-2.6), Blastocystis sp. (1.1%, 6/550; 95% CI: 0.4-2.4) and Encephalitozoon intestinalis (0.2%, 1/550; 95% CI: 0.0-10.1). All samples tested negative for Enterocytozoon bieneusi. Sequence analyses revealed the presence of sub-assemblage BIV (n=1) within G.duodenalis, and Cryptosporidium cuniculus (n=6) and Cryptosporidium andersoni (n=1) within Cryptosporidium. The presence of ruminant-adapted C.andersoni is indicative of a potential cross-species transmission event, although a spurious infection (mechanical carriage) cannot be ruled out. Samples assigned to C.cuniculus belonged to the gp60 subtype families Va (n=3) and Vb (n=2). The six Blastocystis-positive samples were identified as ST2 (n=3) and ST1 + ST2 (n=3). Our molecular results suggest that wild rabbits and hares were primarily infected by leporid-adapted species of eukaryotic pathogens. However, the occasional findings of zoonotic G.duodenalis sub-assemblage BIV, Blastocystis sp. ST1 and ST2, and Encephalitozoon intestinalis could be of public health relevance.

Data for the Domestic Rabbit’s Drinking Water Supply

A short review article was prepared on the domestic rabbit's drinking water consumption and the external and internal factors that significantly influence it. The rabbit's need for drinking water is influenced, among other things, by age, sex, body weight, lactation phase and milk production, the dry matter, fiber and protein content of the feed, the mineral content of the water, the temperature of the water, the possibility of access to water and/or feed and the ambient temperature. The domestic rabbit's water consumption habits are partly derived from the ancestral behavior of the wild rabbit (Oryctolagus cuniculus), and partly influenced by the design of the drinkers. Continuous satisfaction of the rabbit's demand for drinking water in artificial housing conditions is a basic welfare requirement.

Molecular detection of Ehrlichia spp. in ticks parasitizing wild lagomorphs from Spain: characterization of a novel Ehrlichia species

BackgroundSeveral species belonging to the genus Ehrlichia are considered pathogenic for animals and humans. Although wildlife are known to play an important role in the epidemiology of these bacteria, information on the role of wild lagomorphs in their sylvatic cycle is limited. Thus, the objective of the present study was to assess the occurrence of Ehrlichia spp. in ticks collected from wild lagomorphs in Spanish Mediterranean ecosystems.MethodsA total of 1122 pooled ticks (254 pools) collected from 506 wild rabbits (Oryctolagus cuniculus) and 29 Iberian hares (Lepus granatensis) were analysed using a nested PCR assay targeting the partial groEL gene. Ehrlichia spp.-positive samples were further subjected to a second PCR assay targeting 16S rRNA.ResultsThree (1.2%) tick pools comprising Rhipicephalus pusillus collected from nine wild rabbits were positive for Ehrlichia spp. All the Ehrlichia DNA sequences were identical, and use of sequence and phylogenetic analyses allowed us to identify a novel Ehrlichia species.ConclusionsWe provide evidence that a novel Ehrlichia species, named herein as ‘Candidatus Ehrlichia andalusi’, which may be of concern for animal and public health, is circulating in R. pusillus in Spanish Mediterranean ecosystems. Further studies are warranted to assess the epidemiology, pathogenicity and zoonotic potential of this Ehrlichia species.Graphical

Francisella tularensis PCR detection in Cape hares (Lepus capensis) and wild rabbits (Oryctolagus cuniculus) in Algeria

Tularemia is a zoonosis caused by the bacterium Francisella tularensis. Leporids are primary sources of human infections in the northern hemisphere. Africa is classically considered free of tularemia, but recent data indicate that this dogma might be wrong. We assessed the presence of this disease in wild leporids in Algeria. Between 2014 and 2018, we collected 74 leporids carcasses from spontaneously dead or hunted animals. Francisella tularensis DNA was detected by specific real-time PCR tests in 7/36 (19.44%) Cape hares (Lepus capensis) and 5/38 (13.15%) wild rabbits (Oryctolagus cuniculus). Known tularemia arthropod vectors infested half of the PCR-positive animals. At necropsy, F. tularensis-infected animals presented with an enlarged spleen (n = 12), enlarged adrenal glands (12), liver discoloration (12), hemorrhages (11), and pneumonia (11). Immunohistological examination of liver tissue from one animal was compatible with the presence of F. tularensis. Our study demonstrates the existence of tularemia in lagomorphs in Algeria. It should encourage investigations to detect this disease among the human population of this country.

Unravelling the Role of Metabolites for Detecting Physiological State of Wild Animals: European Rabbit's (Oryctolagus cuniculus) Case.

European wild rabbit (Oryctolagus cuniculus) has been defined as a keystone species in the Mediterranean ecosystem. Rabbits have been classed as "endangered" by the IUCN within their native range. In this sense, animal nutrition may play a fundamental and limiting role in the conservation of wild species. The overarching goal of ecological nutrition is to unravel the extensive web of nutritional links that direct animals in their interactions with their ecological environments. The main aim of this work was to evaluate the effect of different feed intake, geographic location, animal sex, and reproductive stage on glucose, non-esterified fatty acids (NEFA), and plasmatic urea nitrogen (PUN), albumin, glutamate, and total protein metabolites. Additionally, we examined the potential of these metabolites as biomarkers. Full stomach contents and blood samples were collected from European wild rabbits (n = 89) for the analysis of the metabolites described above. Our work shows that the levels of these metabolites are affected by the sex of the animals, as well as by their reproductive stage (glucose, NEFA and albumin). There were signs of better optimisation of resources by females than by other groups of animals. These data may be interesting in the study of nutritional components that could be affecting physiological state of this species.

Latrine counts to estimate wild rabbit (Oryctolagus cuniculus) density as a tool for biodiversity conservation and management

The recovery of European wild rabbit (Oryctolagus cuniculus) populations in Spain is of great interest for wildlife conservation. Large-scale management and conservation programmes aimed to determine current population abundance.Most methods for censusing rabbits only offer estimates of relative abundance. However, we lack accurate studies about how latrine counts index is related to population density. The aim of this study is twofold: 1) to test the suitability of counting rabbit latrines as a relative abundance index and; 2) to explore if the number of latrines is a useful predictor of wild rabbit density.We compared latrine surveys conducted in 2 km-length paths with density estimates obtained from distance-based estimates. The study was conducted during two years in central Spain in mixed agriculture-woodland landscapes. Sites covered all potential range of rabbit densities in natural landscapes. Density estimates were obtained by using Distance software.We used type II major axis regression to evaluate the relationship between rabbit density and latrine counting. The predictive ability of the regression equation was performed by several predictive metrics.Results show that the number of latrines counted is a very good abundance index. We also found that latrine counts had a high predictive power to estimate rabbit density. Therefore, latrine surveys are not only a proxy of abundance, but can be used as a cost-effective tool to estimate rabbit population density at low and intermediate densities, which are the most important for the conservation practitioners. Furthermore, the method needs to be tested in other seasons and habitats to prove its generality.