European Wild Rabbit Oryctolagus Research Articles

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.

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.

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.

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.

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.

Long-term response of open-habitats species to wildfire salvage logging: the endangered European wild rabbit as example

Salvage logging treatments, a type of logging to economic returns after natural disturbance, are often applied in the aftermath of wildfires. Specialist or dependent species of open-habitat usually increase their populations in the short-term after wildfires and post-fire salvage logging. However, the long-term effects on threatened open-habitat species such as the European wild rabbit (Oryctolagus cuniculus L.) are still poorly known. Thus, plant productivity, habitat heterogeneity and rabbit abundance were studied in the north-east Iberian Peninsula in four type of post-fire treatment plots: (1) unburnt, (2) salvage logging with branches left on the ground, (3) salvage logging and manual removal of branches, and (4) recurrent fires. Both the time since the fire and the treatment affected plant productivity and habitat heterogeneity. Plant productivity was quicker in treatments when branches were left on the ground or when branches were removed than in recurrent fire plots. Rabbit relative abundance increased in the short term but dramatically declined over time after fires, especially in the plots where branches were left on the ground and with recurrent fires, in which rabbit abundances fell dramatically. In the long-term, the lack of food availability and adequate habitat structure are the main factors affecting the maintenance of the rabbit population. An appropriate moment for managing burnt areas to favour the persistence of rabbit is between the fifth and sixth year after the fire. These actions also benefit the reduction of environmental biomass and so help prevent future severe wildfires.

Nutritional Metabolites as Biomarkers of Previous Feed Intake in European Rabbit (Oryctolagus cuniculus): Applications on Conservation.

Simple SummaryThis work aims to address the use of biomarkers that can provide us with information on the previous nutrition levels of wild rabbits, a keystone species that has drastically reduced in the last few years. Non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites were analysed. Additionally, we examined the potential of these metabolites as biomarkers for the nutritional and conservation status of European rabbits to further the biological knowledge of this species and contribute to its conservation.European wild rabbit (Oryctolagus cuniculus) populations have drastically reduced, and recently, rabbits have been classed as “endangered” by the IUCN. This animal plays an important ecological role in Mediterranean ecosystems and its introduction could significantly contribute to ecological restoration. Rabbits have high nutrient requirements that apparently cannot be covered in all ecosystems, and there are clues that nutrition can limit their abundance and density. On the other hand, some studies reflect the effects of food availability on the metabolomic status of other animal species, but there are no specific studies on this keystone species. The main aim of this work is to find biomarkers to assess the previous levels of ingestion of European rabbits (Oryctolagus cuniculus). To address this gap, gastric content and blood samples were collected from European rabbits (n = 99) in a Mediterranean area for the analysis of glucose, non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites. Depending on their previous feed intake (gastric content and the ratio between the gastric content and the weight of the animal), the animals were divided into two groups (lower and normal previous feed intake). Our work shows that the metabolomic profiles of the animals were affected. Levels of glucose (+82%; p = 0.0003), NEFA (−61%; p = 0.0040) and PUN (+139%; p < 0.001) were different in the animals with lower previous feed intake than the animals with normal previous feed intake. This work summarises that metabolic phenotype can be interesting when seeking to discover the limiting nutrients and food availability in diets that could affect the ecological fitness and conservation of European wild rabbits. It is important to mention that in this work, only the effects on six different metabolites have been analysed and more studies are necessary to complement the knowledge of possible metabolites that indicate the level of ingestion in this species and others. These (and new) biomarkers could be used as a tool to provide information about individual or population characteristics that other physiological parameters cannot detect, improving the conservation physiology field.

Monitoring of Rabbit Hemorrhagic Disease Virus in European Wild Rabbit (Oryctolagus cuniculus) Populations by PCR Analysis of Rabbit Fecal Pellets.

European rabbits (Oryctolagus cuniculus) are affected by rabbit hemorrhagic disease (RHD), which is caused by a lagovirus responsible for significant mortality in European wild rabbit populations. Our study aimed to evaluate the potential for detecting viral RNA by duplex real-time PCR in rabbit fecal pellets collected in the field, as a noninvasive method to monitor RHD virus circulation in wild populations. To do this, monthly discoveries of rabbits that died from RHD and detection of viral RNA in fecal pellets were recorded in two enclosed populations of wild rabbits throughout a year. The results suggested a low performance of this procedure to monitor viral infection incidence and a weak concordance with monthly discoveries of rabbits that died from RHD. This poor association was probably due to the low amount of viral RNA in feces, the prolonged time of excretion after infection, and that the number of rabbits found dead from RHD does not necessarily correlate with RHD incidence. Nevertheless, this procedure may be a complementary noninvasive method to assist in determining the presence of RHD viruses in populations. Additional research is needed to determine the suitability of this methodology to perform epidemiologic surveys on wild populations of European rabbits and, especially, other European or North American lagomorph species affected by lagoviruses.

Co-infection by classic MYXV and ha-MYXV in Iberian hare (Lepus granatensis) and European wild rabbit (Oryctolagus cuniculus algirus).

Myxomatosis is an emergent disease in the Iberian hare (Lepus granatensis). In this species, the disease is caused by a natural recombinant virus (ha-myxoma virus [MYXV]) identified for the first time in 2018 and has since been responsible for a large number of outbreaks in Spain and Portugal. The ha-MYXV, which harbours a 2.8 Kb insert-disrupting gene M009L, can also infect and cause disease in wild and domestic rabbits, despite being less frequently identified in rabbits. During the laboratory investigations of wild leporids found dead in Portugal carried out within the scope of a Nacional Surveillance Plan (Dispatch 4757/17, MAFDR), co-infection events by classic (MYXV) and naturally recombinant (ha-MYXV) strains were detected in both one Iberian hare and one European wild rabbit (Oryctolagus cuniculus algirus). These two cases were initially detected by a multiplex qPCR detection of MYXV and ha-MYXV and subsequently confirmed by conventional PCR and sequencing of the M009L gene, which contains an ha-MYXV-specific insertion. To our knowledge, this is the first documented report of co-infection by classic MYXV and ha-MYXV strains either in Iberian hare or in European wild rabbit. It is also the first report of infection of an Iberian hare by a classic MYXV strain. These findings highlight the continuous evolution of the MYXV and the frequent host range changes that justify the nonstop monitoring of the sanitary condition of wild Leporidae populations in the Iberian Peninsula.

Changes in European wild rabbit population dynamics and the epidemiology of rabbit haemorrhagic disease in response to artificially increased viral transmission

European wild rabbit (Oryctolagus cuniculus) populations are severely affected by rabbit haemorrhagic disease (RHD), currently aggravated by the spread of the new lagovirus serotype RHDV2 that replaced the classical RHDV strains (RHDV/RHDVa). This virus causes high mortality in both adult and young rabbits and to date, there is no management tool to effectively reduce its impact on wild rabbit populations. This hinders the success of common strategies, such as habitat management or restocking, in areas where rabbits are native. However, the present study, conducted on enclosed wild rabbit populations, showed that spreading RHDV2 on baits during breeding periods induced infection of young rabbits, reducing mortality rates, presumably due to maternal antibody protection. This reduced the young rabbit mortality hazard by a third, and more juvenile rabbits immune to RHDV2 were recruited into the adult breeding population. Young rabbits from populations in which the force of infection of RHDV2 was increased, however, exhibited considerably higher susceptibility to infection by RHDV than those from non-treated control populations. Since co-circulation of classical RHDVs was ruled out, differences in the type and degree of immunization, the level of cross-protection and/or other unknown factors, such as the circulation of undetected non-pathogenic lagoviruses, arose as possible explanations. This meant that although the present study demonstrated the possibility of successfully modulating the impact of RHD in wild populations, the epidemiological complexity of the situation where several lagoviruses circulate requires additional research to determine final applicability of the proposed method.

Molecular identification of zoonotic Rickettsia species in Ixodidae parasitizing wild lagomorphs from Mediterranean ecosystems.

A survey study was carried out to identify tick species parasitizing wild lagomorphs in Mediterranean ecosystems in southern Spain and to determine the occurrence of Rickettsia species present in these ticks in this region. A total of 1304 European wild rabbits (Oryctolagus cuniculus) and 58 Iberian hares (Lepus granatensis) were individually examined for the presence of ticks. Ticks were found in 42.9% and 50% of the wild rabbits and hares sampled, respectively. A total of 1122 ticks were collected and five species, including Rhipicephalus pusillus, Hyalomma lusitanicum, Haemaphysalis hispanica, Ixodes ventalloi and Rhipicephalus sanguineus sensu lato (s.l.), were microscopically and molecularly identified at the 16S rRNA gene. This is the first study on Ixodidae parasitizing Iberian hares. The presence of Rickettsia DNA was assessed in 254 tick pools (according to hunting states, lagomorph species, tick species and tick development stage) using PCR assays targeting the rOmpA, rOmpB and gltA. Twenty-seven pools (10.6%) were positive to Rickettsia DNA. Five zoonotic Rickettsia species were identified, being Rickettsia massiliae the most frequent (4.7%), followed by Rickettsia sibirica subsp. mongolitimonae (2.8%), Rickettsia slovaca (2.0%), Rickettsia aeschlimannii (0.8%) and Rickettsia africae (0.4%). The results suggest that wild rabbits and Iberian hares are parasitized by a wide range of tick species and that these lagomorphs may play an important role in the sylvatic cycle of some zoonotic Rickettsia species in Mediterranean ecosystems. Our data represent the first report of R. massiliae, R. aeschlimannii, R. slovaca and R africae in ticks collected in wild lagomorphs in Europe, and the first report of not imported R. africae in this continent. Since R. slovaca and R. africae DNA was detected in tick species different to their main vectors, further studies are warranted to unravel the role of wild lagomorphs in the epidemiology of these vector-borne pathogens.

What Are the Keys to the Adaptive Success of European Wild Rabbit (Oryctolagus cuniculus) in the Iberian Peninsula?

Simple SummaryWhy might a species both be seriously threatened and pose an overpopulation problem in introduced locations? The aim of this review was to understand the keys to the adaptive success of the wild rabbit (Oryctolagus cuniculus) in order to establish its strengths and weaknesses for the management of this keystone species in Mediterranean ecosystems. This work highlights the need to create specific conservation programs for this species.The European wild rabbit (Oryctolagus cuniculus) plays an important ecological role in the ecosystems of the Iberian Peninsula. Recently, rabbit populations have drastically reduced, so the species is now considered endangered. However, in some places, this animal is considered a pest. This is the conservation paradox of the 21st century: the wild rabbit is both an invasive alien and an endangered native species. The authors of this review aimed to understand the keys to the adaptive success of European rabbits, addressing all aspects of their biology in order to provide the keys to the ecological management of this species. Aspects including nutrition, genetics, immunity interactions with the environment, behaviour, and conflict with human activities were reviewed. Ultimately, rabbits are resilient and adaptable. The main adaptations that explain the rabbit’s adaptive success are its nutrition (wide adaptation to food and good nutritional use of caecotrophy), immune system (powerful and developed), and other aspects related to genetics and behaviour. Rabbits’ relationship with humans has led them to colonise other places where they have become pests. Despite these adaptations, populations in native places have been drastically reduced in recent years. Since it serves as a bastion of the Mediterranean ecosystem, a specific conservation program for this species must be carried out. Therefore, a study of the rabbit’s response to diseases and nutrition (especially protein), as well as the interaction between them, is of special interest.

Persistence of wild rabbit (Oryctolagus cuniculus) latrines and its implication for monitoring programs

Counting latrines is a standard method to estimate European wild rabbits (Oryctolagus cuniculus) abundance. This method plays an important role in endangered predators’ conservation programs in the Iberian Peninsula, especially in potential reintroduction areas for the Iberian lynx (Lynx pardinus). However, since the last half-century, rabbit populations have declined dramatically within their native range, mainly due to the effect of two viral diseases, which have caused even local extinctions of natural populations. Hence, we raise the question regarding how reliable the method of counting latrines may be in ensuring the abundance of rabbit populations after an event of sudden decline or even local extinction. To answer this question, we carry out a simple experiment where we set ten lines with rabbit latrines with two different sizes each (small and large) and simulated low- and high-abundant rabbit population scenarios on five latrine lines each, respectively. Then, we calculated the pellets decay rate and latrines persistence rate through monthly surveys. Our results showed that the daily persistence rate varied little among the different scenarios. However, generally, latrines do not persist long (78 days on average) regardless of the size of the latrines or rabbit population abundance. The longest time to extinction for pellets (c. 469 days, 97.5 quantile) was longer than the time until no more latrines were identified (c. 191 days, 97.5 quantile). We conclude that, although we did not consider other factors that could affect pellets decay, counting latrines seems to be sensible to a hypothetical dramatic decline event of a wild rabbit population. Therefore, if there is evidence of an epidemiological outbreak event, we suggest managers carry out latrines census at least between 140–220 days after an outbreak detection to reduce bias on the determination of rabbit abundance.

Ectoparasites of wild rabbit (Oryctolagus cuniculus) in Southern Italy

The European wild rabbit Oryctolagus cuniculus is a widespread mammal that may act as host for ectoparasites and reservoir for some vector-borne pathogens. Study aims were to investigate the arthropod fauna that may infect the European wild rabbit, to assess the ecological indexes and to investigate tick preferred attachment sites. The survey was conducted on 105 wild rabbits hunted by poachers in a Mediterranean woodland area in the province of Ragusa. Animals were individually examined for ectoparasite presence and the tick load assessed on eleven body anatomical sites. Ticks and fleas were found on 79 (75.2%) and 3 (2.9%) out of the 105 examined rabbits, respectively. Three tick species were identified; Rhipicephalus pusillus was the most abundant followed by Rhipicephalus turanicus and Ixodes gibbosus. Although ticks were found on all anatomical sites, ears, forelimbs, and abdomen were those more frequently infected. All the collected fleas were identified as Spilopsyllus cuniculi. Some of the ectoparasite species found on examined wild rabbits are recognized as potential vectors of pathogens of animal and human concern, such as, Rickettsia sibirica mongolitimonae and Rickettsia massiliae. The most abundant tick species found in this study is the main vector of some zoonotic pathogens; therefore, the possible role of O. cuniculus in their maintenance should be further investigated.

Detection of Myxoma Virus DNA in Ticks from Lagomorph Species in Spain Suggests Their Possible Role as Competent Vector in Viral Transmission.

Myxoma virus (MYXV) causes morbidity and mortality in European wild rabbits (Oryctolagus cuniculus) worldwide, and recently in Iberian hares (Lepus granatensis) in Spain. We aimed to assess the presence of MYXV-specific DNA in ixodid ticks collected from both hosts. A total of 417 ticks harvested from 30 wild lagomorphs, including wild rabbits and Iberian hares were collected from southern Spain. Enzyme-linked immunosorbent assay and PCR-sequencing were used to detect virus exposure and presence, respectively. Antibodies to MYXV were detected in 68% (17/25) of wild rabbits and in 67% (2/3) of Iberian hares. We detected MYXV DNA in 50.7% of pools of two different tick species (nymphs and adults of Rhipicephalus pusillus, and nymphs of Hyalomma lusitanicum) parasitizing rabbits and hares. The obtained partial sequence of the viral major envelope protein gene showed a mutation (G383A) within the MYXV_gp026 locus between the rabbit strain and Iberian hare strain (recently isolated in tissues of infected hares from Spain). However, in our study, the viral DNA presence was detected for the first time using tick DNA as the PCR-template, but the possible role of ticks as vectors of MYXV still needs to be elucidated.

PCR Detection of Toxoplasma gondii in European Wild Rabbit (Oryctolagus cuniculus) from Portugal.

Wildlife plays an important role in the epidemiological cycle of Toxoplasma gondii. The European wild rabbit (Oryctolagus cuniculus) can be a source of infection to wild and domestic hosts, including human beings. Additionally, as an herbivorous animal, the European wild rabbit may also be a sentinel of environmental contamination with T. gondii and, consequently, an indicator of the potential transmission of this parasite. The purpose of the present work was to detect T. gondii DNA in European wild rabbit from central Portugal, as well as the possible implications for public health. Heart and diaphragm samples were obtained from 28 rabbits hunted in central Portugal. Nested PCR separately amplified the 5′ and 3′ ends of the surface antigen 2 (SAG2) gene. T. gondii DNA was detected in 19 out of the 28 sampled animals, resulting in a prevalence of 67.9%. These results show that T. gondii infection occurs in European wild rabbit and therefore may pose a potential risk for humans if consumed as raw or undercooked meat. Measures should be taken in order to prevent infection by this zoonotic parasite and for the conservation of wildlife. To the best of our knowledge, this is the first study performed by means of PCR on T. gondii in European wild rabbit meat samples.

Zoonotic Microsporidia in Wild Lagomorphs in Southern Spain.

Simple SummaryA cross-sectional study was carried out to assess the presence of zoonotic microsporidia in organ meats of European wild rabbits and Iberian hares consumed by humans in Spain. Between July 2015 and December 2018, kidney samples from 383 wild rabbits and kidney and brain tissues from 79 Iberian hares in southern Spain were tested by species-specific polymerase chain reactions (PCRs) for the detection of microsporidia DNA. We confirmed the presence of Enterocytozoon bieneusi in three wild rabbits and Encephalitozoon intestinalis in one wild rabbit and three Iberian hares. However, none of the 462 sampled wild lagomorphs showed Encephalitozoon hellem nor Encephalitozoon cuniculi infection. This is the first report of E. intestinalis infection in wild rabbits and Iberian hares. The presence of E. bieneusi and E. intestinalis in organ meats from wild lagomorphs can be of public health concern. Additional studies are required to determine the real prevalence of these parasites in European wild rabbit and Iberian hare.Microsporidia are obligate intracellular protist-like fungal pathogens that infect a broad range of animal species, including humans. This study aimed to assess the presence of zoonotic microsporidia (Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi) in organ meats of European wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis) consumed by humans in Spain. Between July 2015 and December 2018, kidney samples from 383 wild rabbits and kidney and brain tissues from 79 Iberian hares in southern Spain were tested by species-specific PCR for the detection of microsporidia DNA. Enterocytozoon bieneusi infection was confirmed in three wild rabbits (0.8%; 95% CI: 0.0–1.7%) but not in hares (0.0%; 95% CI: 0.0–4.6%), whereas E. intestinalis DNA was found in one wild rabbit (0.3%; 95% CI: 0.0–0.8%) and three Iberian hares (3.8%; 95% CI: 0.0–8.0%). Neither E. hellem nor E. cuniculi infection were detected in the 462 (0.0%; 95% CI: 0.0–0.8%) lagomorphs analyzed. The absence of E. hellem and E. cuniculi infection suggests a low risk of zoonotic foodborne transmission from these wild lagomorph species in southern Spain. To the authors’ knowledge, this is the first report of E. intestinalis infection in wild rabbits and Iberian hares. The presence of E. bieneusi and E. intestinalis in organ meats from wild lagomorphs can be of public health concern. Additional studies are required to determine the real prevalence of these parasites in European wild rabbit and Iberian hare.