Comparison of biological features of wild European rabbit mesenchymal stem cells derived from different tissues.

Although the European rabbit is an "endangered" species and a notorious biological model, the analysis and comparative characterization of new tissue sources of rabbit mesenchymal stem cells (rMSCs) has not been well studied. Here we report for the first time the isolation and characterization of rMSCs derived from an animal belonging to a natural rabbit population within the species native region. New rMSC lines were isolated from different tissues: oral mucosa (rOM-MSC), dermal skin (rDS-MSC), subcutaneous adipose tissue (rSCA-MSC), ovarian adipose tissue (rOA-MSC), oviduct (rO-MSC), and mammary gland (rMG­MSC). The six rMSC lines showed plastic adhesion with fibroblast-like morphology and were all shown to be positive for CD44 and CD29 expression (characteristic markers of MSCs), and negative for CD34 or CD45 expression. In terms of pluripotency features, all rMSC lines expressed NANOG, OCT4, and SOX2. Furthermore, all rMSC lines cultured under osteogenic, chondrogenic, and adipogenic conditions showed differentiation capacity. In conclusion, this study describes the isolation and characterization of new rabbit cell lines from different tissue origins, with a clear mesenchymal pattern. We show that rMSC do not exhibit differences in terms of morphological features, expression of the cell surface, and intracellular markers of pluripotency and in vitro differentiation capacities, attributable to their tissue of origin.

A Quadruplex qPCR for Detection and Differentiation of Classic and Natural Recombinant Myxoma Virus Strains of Leporids

A natural recombinant myxoma virus (referred to as ha-MYXV or MYXV-Tol08/18) emerged in the Iberian hare (Lepus granatensis) and the European rabbit (Oryctolagus cuniculus) in late 2018 and mid-2020, respectively. This new virus is genetically distinct from classic myxoma virus (MYXV) strains that caused myxomatosis in rabbits until then, by acquiring an additional 2.8 Kbp insert within the m009L gene that disrupted it into ORFs m009L-a and m009L-b. To distinguish ha-MYXV from classic MYXV strains, we developed a robust qPCR multiplex technique that combines the amplification of the m000.5L/R duplicated gene, conserved in all myxoma virus strains including ha-MYXV, with the amplification of two other genes targeted by the real-time PCR systems designed during this study, specific either for classic MYXV or ha-MYXV strains. The first system targets the boundaries between ORFs m009L-a and m009L-b, only contiguous in classic strains, while the second amplifies a fragment within gene m060L, only present in recombinant MYXV strains. All amplification reactions were validated and normalized by a fourth PCR system directed to a housekeeping gene (18S rRNA) conserved in eukaryotic organisms, including hares and rabbits. The multiplex PCR (mPCR) technique described here was optimized for Taqman® and Evagreen® systems allowing the detection of as few as nine copies of viral DNA in the sample with an efficiency > 93%. This real-time multiplex is the first fast method available for the differential diagnosis between classic and recombinant MYXV strains, also allowing the detection of co-infections. The system proves to be an essential and effective tool for monitoring the geographical spread of ha-MYXV in the hare and wild rabbit populations, supporting the management of both species in the field.

Breeding of Cory's shearwater Calonectris borealis on Selvagem Grande and beneficial effects of removal of invasive mammals

The largest colony of Cory's shearwater Calonectris borealis nests on the island of Selvagem Grande in the north-eastern Atlantic. In 2002, a programme of eradication was conducted to remove two alien invasive mammals, the house mouse Mus musculus and European rabbit Oryctolagus cuniculus. Preliminary studies recorded beneficial effects of the eradications for a variety of plant and animal species, including Cory's shearwater. We recorded fledging rates of shearwaters for 1982–2001, prior to the eradication, and for 2002–2020, after the eradication, from two quadrats, each containing 134–329 nest sites. Although there was annual fluctuation in fledging rates in the quadrats, the mean rate of 40.74 ±SD 3.92 fledglings per 100 nest sites for the two quadrats combined prior to the eradication of mammals increased significantly, to 52.88 ± SD 5.03 per 100 nest sites, after the eradications. Because the two mammals were removed synchronously it is difficult to know which factors depressed fledging of Cory's shearwaters on Selvagem Grande. However, the predatory behaviour of house mice on other oceanic islands, and the fact that increased fledging was seen soon after the eradications occurred, suggest predation by house mice on shearwater hatchlings was the main cause of losses.

Temporal patterns of wildlife roadkill in the UK

Wildlife-vehicle collisions are one of the main causes of mortality for wild mammals and birds in the UK. Here, using a dataset of 54,000+ records collated by a citizen science roadkill recording scheme between 2014–2019, we analyse and present temporal patterns of wildlife roadkill of the 19 most commonly reported taxa in the UK (84% of all reported roadkill). Most taxa (13 out of 19) showed significant and consistent seasonal variations in road mortality and fitted one of two seasonal patterns; bimodal or unimodal: only three species (red fox Vulpes vulpes, European polecat Mustela putorius and Reeves’ muntjac deer Muntiacus reevesi) showed no significant seasonality. Species that increase movement in spring and autumn potentially have bimodal patterns in roadkill due to the increase in mate-searching and juvenile dispersal during these respective time periods (e.g. European badger Meles meles). Unimodal patterns likely represent increased mortality due to a single short pulse in activity associated with breeding (e.g. birds) or foraging (e.g. grey squirrels Sciurus carolinensis in autumn). Importantly, these patterns also indicate periods of increased risk for drivers, potentially posing a greater threat to human welfare. In addition to behaviour-driven annual patterns, abiotic factors (temperature and rainfall) explained some variance in roadkill. Notably, high rainfall was associated with decreased observations of two bird taxa (gulls and Eurasian magpies Pica pica) and European rabbit Oryctolagus cuniculus. By quantifying seasonal patterns in roadkill, we highlight a significant anthropogenic impact on wild species, which is important in relation to conservation, animal welfare, and human safety.

Viral Disease in Lagomorphs: A Molecular Perspective

Our understanding of molecular biology of the viruses that infect lagomorphs is largely limited to the leporipoxvirus myxoma virus (MYXV) and the lagoviruses rabbit haemorrhagic disease virus (RHDV) and European brown hare syndrome virus (EBHSV) that infect the European rabbit (Oryctolagus cuniculus) and the European brown hare (Lepus europaeus) respectively. Thanks to the great effort of historic surveillance studies and careful sample archiving, the molecular evolution of these viruses is being resolved. Although historically considered viruses that cause species specific diseases recent reports show that several lagomorphs may now face the threat of these maladies. The driving factors behind these changes has not been determined and the effect of these species jumps on lagomorph populations has yet to be seen. Lagomorphs are also affected by several other lesser studied viral diseases. In addition, recent metagenomic studies have led to the identification of novel lagomorph viruses the importance of these to lagomorph health remains to be fully determined. In this chapter we summarize molecular aspects of viruses that infect lagomorphs, paying particular attention to recent interspecies infections.

Craniometric Analysis of European Rabbit (Oryctolagus Cuniculus) Breeds to Trace out Intraspecific and Inter Gender Morphometric Variations

The family Liproidae expresses the significant variation of morphometric features at species level. In already available literature on the European rabbit (Oryctolagus cuniculus) the idea is proposed that there can be variation in the morphometric character of different breeds of Oryctolagus cuniculus but there are negligible scientific research base evidences for it. So in this study interbreed craniometric comparison of the European rabbit is carried out to trace out their intergender and intraspecific morphometric variations. The study material consists of 32 sexually mature rabbits belong to two breeds of European rabbit that are New Zealand rabbit (eight males and eight females) and American Dutch rabbit (eight males and eight females) collected from different districts of the Punjab, Pakistan. There were 29 different measurements taken from the skull and mandible of both breeds by using digital and manual verniar calipers. The results of this study have revealed that there is significant interbreed variation in the nasal length measurement of the New Zealand and American Dutch rabbit. This significant interbreed difference in nasal length is observed in both female as well as male rabbits (p=0.0059 and 0.0069 respectively). The current study also depicts that inter gender craniometric variations exist in the New Zealand as well as American Dutch rabbit. The standard deviation of the foramen magnum index of the both included breeds is comparatively higher than all the other included morphometric features. This study provides the baseline data on interbreed craniometric variations in the Oryctolagus cuniculus.

Dynamic Molecular Evolution of Mammalian Homeobox Genes: Duplication, Loss, Divergence and Gene Conversion Sculpt PRD Class Repertoires

The majority of homeobox genes are highly conserved across animals, but the eutherian-specific ETCHbox genes, embryonically expressed and highly divergent duplicates of CRX, are a notable exception. Here we compare the ETCHbox genes of 34 mammalian species, uncovering dynamic patterns of gene loss and tandem duplication, including the presence of a large tandem array of LEUTX loci in the genome of the European rabbit (Oryctolagus cuniculus). Despite extensive gene gain and loss, all sampled species possess at least two ETCHbox genes, suggesting their collective role is indispensable. We find evidence for positive selection and show that TPRX1 and TPRX2 have been the subject of repeated gene conversion across the Boreoeutheria, homogenising their sequences and preventing divergence, especially in the homeobox region. Together, these results are consistent with a model where mammalian ETCHbox genes are dynamic in evolution due to functional overlap, yet have collective indispensable roles.

Feral rabbit populations in a peri-urban area: insights about invasion dynamics and potential management strategies

Where introduced, the European rabbit Oryctolagus cuniculus is considered among the most destructive and invasive species. To date, research focused mostly on populations of wild rabbit, whereas little is known on feral domestic rabbit populations. In this work, we reported the establishment of two self-sustaining populations of feral rabbits in Italy. Direct observations were conducted to assess rabbit range expansion and population increase over time. We also evaluated prey-predator interactions between rabbits and native red foxes Vulpes vulpes, by means of camera trapping and the analysis of fox scats. Moreover, we also assessed the social perception towards feral rabbits and the acceptability of various management options through the administration of a structured questionnaire to park visitors. Rabbit populations increased between 2018 and 2019, as well as the size of the invaded range. Rabbits are predated by foxes, but they seem to have adapted their activity rhythms to minimize the risk of predation, becoming diurnal. Park visitors loved rabbits and deemed them to be a positive presence in the park, which deserve to live there. Surgical sterilization of rabbits was the only management option which was partially supported. Feral domestic rabbits, albeit a neglected species in invasion biology, can become extremely successful invaders of urban green areas: in < 5 years, rabbits were able to colonize our study area and become a food resource for native predators and also an iconic species. These three points raise concerns about the potential impacts of invasive feral rabbits in European urban green areas and call for further research in this direction.

A Review on the Methods Used for the Detection and Diagnosis of Rabbit Hemorrhagic Disease Virus (RHDV)

Since the early 1980s, the European rabbit (Oryctolagus cuniculus) has been threatened by the rabbit hemorrhagic disease (RHD). The disease is caused by a lagovirus of the family Caliciviridae, the rabbit hemorrhagic disease virus (RHDV). The need for detection, identification and further characterization of RHDV led to the development of several diagnostic tests. Owing to the lack of an appropriate cell culture system for in vitro propagation of the virus, much of the methods involved in these tests contributed to our current knowledge on RHD and RHDV and to the development of vaccines to contain the disease. Here, we provide a comprehensive review of the RHDV diagnostic tests used since the first RHD outbreak and that include molecular, histological and serological techniques, ranging from simpler tests initially used, such as the hemagglutination test, to the more recent and sophisticated high-throughput sequencing, along with an overview of their potential and their limitations.