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Nesolagus netscheri, a Sumatran striped rabbit, is one of the rarest rabbits in the Leporidae family, and its genetic information is still limited. This study provides the first mitochondrial genome and molecular systematic characterization of the Sumatran striped rabbit, Nesolagus netscheri, Indonesia's rarest rabbit. It consists of a circular double-stranded DNA of 16,709 bp. It showed that the mitochondrial genome structure of N. netscheri is similar to that of N. timminsi. The mitochondrial genome of N. netscheri contained 22 transfer RNA (tRNA) genes, and all tRNA except for trnS1 showed a characteristic cloverleaf secondary structure. Evidence was found that the atp8 gene of N. netscheri is under positive selection pressure. The phylogenetic analysis shows Leporidae was monophyletic, with Nesolagus at the basal. The study indicates a split between N. netscheri and N. timminsi in the Late Pleistocene around 0.43 million years ago. This research is a fundamental reference for the conservation of the rarest lagomorph species and provides important information for future evolutionary studies in the Leporidae family., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

An 8-year-old rabbit presented with a 5-day history of acute difficulty in walking. Neurological examination revealed tetraparesis, proprioceptive deficits in both pelvic limbs and the right thoracic limb, decreased withdrawal reflex on the right thoracic limb and hyperreflexia in the pelvic limbs. A cervico-thoracic (C6-T2) localization was suspected. Computer tomography (CT) and magnetic resonance imaging (MRI) scans were performed, revealing a right dorsolateral extradural lesion at the C6-C7 intervertebral disc space. Additionally, meningeal and paravertebral contrast enhancement was observed on MRI, while periosteal reaction was evident at the right C6-C7 facet joint on CT. The findings were primarily consistent with spinal cord compression due to the presence of extruded disc material. Following conservative treatment failure, a right-sided C6-C7 hemilaminectomy was performed to remove the compression and sample the extradural material. Histological examination confirmed the presence of degenerated and partially mineralized disc material mixed with granulation tissue. This is the first reported case of cervical disc extrusion in a rabbit, confirmed by histological examination.

The internal carotid artery (ICA) is one of the major vessels in the cranial circulation. Characters concerning the ICA, such as its course in the auditory region, have been employed frequently in phylogenetic analyses of mammals, including extinct taxa. In lagomorphs, however, our knowledge on vascular features of the auditory region has been based predominantly on living species, mostly on the European rabbit. We present the first survey on 11 out of 12 extant genera and key fossil taxa such as stem lagomorphs and early crown representatives ( Archaeolagus and Prolagus ). The ICA pattern shows a modified transpromontorial course in stem taxa ( Litolagus , Megalagus and Palaeolagus ) and Archaeolagus , which we propose as the ancestral character state for Lagomorpha, similar to that for the earliest rodents, plesiadapids and scandentians. The ICA pattern in leporids is perbullar, but shows structural similarities to stem taxa, whereas the extrabullar ICA course in Ochotona is apparently a highly derived condition. Prolagus shows a mixed character state between leporids and Ochotona in its ICA route. The persistence of the transpromontorial ICA course and similarities in the carotid canal structure among stem taxa and crown leporids support morphological conservatism in Lagomorpha, in contrast to their sister clade Rodentia. This article is part of the theme issue 'The mammalian skull: development, structure and function'.

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., (© 2022 Wiley-VCH GmbH.)

The mountain hares ( Lepus timidus L., 1758) in the Faroe Islands, an archipelago located in the North Atlantic, are known to be commonly infected by tapeworms, the identity of which was unknown. The mountain hare, which now populates 15 of the 18 islands, was introduced from Norway in 1855. In this study, tapeworms collected from four mountain hares from four geographic areas of the Faroe Islands were subjected to molecular identification using the nuclear ribosomal DNA (28S), the mitochondrial cytochrome oxidase subunit 1 ( cox1 ) and the NADH dehydrogenase subunit 1 ( nad1 ) genes. The results indicate unambiguously that the tapeworms were Mosgovoyia pectinata (Goeze, 1782) (Cestoda: Anoplocephalidae sensu stricto ). The phylogenetic position and origin of the Faroese M. pectinata are discussed. Given that the parasite is quite common in Norway, from where the mountain hares were introduced, it is conceivable that co-introduction of M. pectinata from Norway to the Faroe Islands took place. The phylogenetic analyses revealed high similarity of the M. pectinata sequences from three regions and the position of the Faroese isolate as the sister lineage of the isolates from Finland and East Siberia., Competing Interests: None., (© 2023 The Authors.)

The presence of microplastics (MPs) in marine environments has been extensively documented. However, studies of terrestrial species are scarce. Fecal samples (105) of lagomorphs were collected at sites with different levels of urbanization in the Baja California Chaparral and analyzed to quantify and characterize MPs found in the feces. The lagomorph species recorded in the study area are the desert cottontail rabbit (Sylvilagus audubonii), brush rabbit (Sylvilagus bachmani), and black-tailed jackrabbit (Lepus californicus), which play important roles in the food web of the chaparral ecosystem. Microplastics were identified using attenuated total reflectance Fourier transform infrared spectroscopy. Microplastics were detected in 49 % of the samples, with fibers being the dominant shape found (72 %). Most (75 %) of the MPs were <1 mm in size, with a mean length of 0.93 ± 0.99 mm (median 0.60 mm, range 0.02 - <5 mm). Polyamide was the dominant polymer (54 %), indicating that MPs are likely derived from textiles; polyethylene was also abundant (27 %). A difference was also observed in the abundance of MPs in feces from sites with different levels of urbanization, with the highest abundance in feces from the urban sites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: The Tawny owl ( Strixaluco ) is a common owl species in Europe, demonstrating generalist diet strategy. Its main prey are small rodents and numerous studies show that the composition of its diet broadly reflects changes in prey species abundance in its habitats. Due to strictly sedentary habits of adult birds and their year-round territoriality, it is possible to locate habitats of their prey with a precision of several hundred metres. Analysis of owl pellets is a traditional method in faunistic studies to provide data on distribution of small mammals, especially cryptic species which are hard to be found using trapping., New Information: Here, we present a dataset on mammals found in Tawny owl pellets collected during up to 13-year studies in the Kharkiv Region, Ukraine in three territories. Data from two territories were collected in a systematic way and allowed us to make analysis of seasonal, year-to-year and habitat variability in the Tawny owl diet and local mammal species composition., (Yehor Yatsiuk, Nataliia Brusentsova, Yuliya Filatova.)

The Amami rabbit, Pentalagus furnessi (Mammalia: Lagomorpha: Leporidae), is a relict and endangered species endemic to the Amami-Oshima and Tokunoshima Islands, located in southwestern Japan. Here, we described three new species of Eimeria (Apicomplexa: Eimeriidae) parasites detected from fecal samples of wild Amami rabbits. Eimeria furnessi n. sp., recorded in 21 (58.3%) samples, has ellipsoidal oocysts with two walls and micropyle, 26.0 × 16.6 μm, and elongate-ovoidal sporocysts, 13.1 × 6.3 μm, with Stieda body. Eimeria hilleri n. sp., recorded in 9 (25.0%) samples, has ellipsoidal oocysts with two walls and micropyle, 34.7 × 21.4 μm, and elongate-fusiform to elongate-ovoidal sporocysts, 15.7 × 8.3 μm, with Stieda and substieda bodies. Eimeria sagentae n. sp., recorded in 13 (36.1%) samples, has ellipsoidal oocysts with two walls and micropyle, 20.9 × 14.5 μm, and elongate-ovoidal sporocysts, 10.4 × 5.0 μm, with Stieda body. The three new species can be distinguished by the size and color of their oocysts. Further studies related to the pathogenicity of these parasites can improve the breeding and propagation procedures of the Amami rabbit., Competing Interests: No conflict of interest declared by any author., (© 2022 The Authors.)

Background: The 'Zacatuche', 'Teporingo', or Volcano rabbit (Romerolagus diazi) belongs to the family Leporidae, is an endemic species restricted to the Central part of the Trans-Mexican Volcanic Belt, and is considered 'endangered' by the IUCN Red List of Threatened Species., Methods and Results: This study reports, for the first time, the complete mitochondrial genome of R. diazi and examined the phylogenetic position of R. diazi among other closely related co-familiar species using mitochondrial protein-coding genes (PCGs). The mitogenome of R. diazi was assembled from short Illumina 150 bp pair-end reads with a coverage of 189x. The AT-rich mitochondrial genome of R. diazi is 17,400 bp in length and is comprised of 13 PCGs, two ribosomal RNA genes, and 22 transfer RNA genes. The gene order observed in the mitochondrial genome of R. diazi is identical to that reported for other leporids. Phylogenetic analyses based on PCGs support the basal position of Romerolagus within the Leporidae, at least when compared to the genera Oryctolagus and Lepus. Nonetheless, additional mitochondrial genomes from species belonging to the genera Bunolagus, Sylvilagus, and Pronolagus, among others, are needed before a more robust conclusion about the derived vs basal placement of Romerolagus within the family Leporidae can be reached based on mitochondrial PCGs., Conclusions: This is the first genomic resource developed for R. diazi and it represents a tool to improve our understanding about the ecology and evolutionary biology of this iconic and endangered species., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal-epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated.

Background: Rabbit hemorrhagic disease virus (RHDV, Lagovirus europeus GI.1) induces a contagious and highly lethal hemorrhagic disease in rabbits. In 2010 a new genotype of lagovirus (GI.2), emerged in Europe, infecting wild and domestic population of rabbits and hares., Case Presentation: We describe the infection with a GI.2 strain, "Bremerhaven-17", in captive mountain hares (Lepus timidus) in a zoo facility in Germany. Postmortem examination revealed RHD-like lesions including necrotizing hepatitis. RT-qPCR and AG-ELISA confirmed presence of GI.2. Recombination and phylogenetic analysis grouped the identified strain with other GI.2 strains, sharing nucleotide identity of 91-99%., Conclusion: Our findings confirm that mountain hares are susceptible to GI.2 infection, due to a past recombination event facilitating virus spillover from sympatric rabbits.

Viruses are ubiquitous in nature; however, very few have been identified in the Leporid species. In the fall of 2018, an outbreak of myxomatosis in Iberian hares ( Lepus granatensis ) was reported in Spain and a novel recombinant myxoma virus strain (MYXV-Tol) was identified. To investigate variability within the recombinant region of the MYXV-Tol and identify any potential viral coinfections, samples (ear, eyelid or vaginal) of Iberian hares were collected from Spain and analyzed. The presence of the recombinant region of the MYXV-Tol was confirmed in six out of eleven samples analyzed. Additionally, a polyomavirus (family Polyomaviridae ), representing a putative new species, and anelloviruses (family Anelloviridae ) belonging to two putative species were identified, some as coinfection with the recombinant MYXV-Tol. The two polyomavirus genomes were identified in two hares and share >99% genome-wide identity. Based on the analysis of their large T-antigen, the new polyomavirus clusters in a distant clade from other mammals sharing <64% amino acid identity. A total of 14 anelloviruses were identified, which share 63-99% genome-wide identity. Overall, our results show a coinfection of different DNA viruses in the studied samples and raise awareness regarding the extensive unsampled diversity of viruses in hares.

Excavations at Kanapoi in north-western Kenya have yielded the most numerically abundant and taxonomically diverse early Pliocene (4.19 Ma) terrestrial small mammal assemblage known from Kenya. A minimum of 15 species are reported, including soricids, sengis, leporids, and rodents: all taxa are referable to extant genera, with the exception of the murine rodent, Saidomys. The majority of the terrestrial small mammals are derived from a bone bed at Nzube's Mandible Site, closely associated with the holotype mandible of Australopithecus anamensis. A smaller number were surface-collected or obtained from screening at several other sites, including the Bat Site. Most small mammals from Nzube's Mandible Site and the Bat Site likely represent prey accumulated as regurgitated pellets from owls, in particular barn owls or giant eagle owls. The small mammal fauna is dominated by the spiny mouse, Acomys: the next most commonly recovered taxa are the multimammate mouse, Mastomys, and the African gerbil, Gerbilliscus. Comparisons of the Kanapoi fauna to other eastern African late Miocene-Pliocene (and one Pleistocene) faunas at the generic level suggest the greatest similarity is to Lemudong'o, Kenya, and Omo B and Aramis, Ethiopia. Further similarities with other localities such as Laetoli, Tanzania, and Hadar, Ethiopia, suggest the existence of a corridor for dispersal along the East African Rift Valley between Ethiopia, Kenya, and Tanzania in the early Pliocene. Further comparisons of the relative abundances of individuals in different families (or subfamilies) emphasize the distinctiveness of the Kanapoi small mammal fauna. The Kanapoi fauna is likely derived from a heterogeneous but relatively arid environment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Leishmania infantum infection in wildlife is increasingly reported in Europe, but scant data are available in Italy so far. This study aimed to investigate the circulation of L. infantum among sylvatic hosts in Sicily (southern Italy), a highly endemic area for canine leishmaniosis, through serological and molecular tools. Target tissues (skin, spleen, lymph nodes) collected from 71 European rabbits, 2 European hares, 7 red foxes, 11 European wildcats and 1 pine marten, were qPCR analysed for the detection of L. infantum DNA. Additionally, 40 rabbits, older than one year, were serologically screened for specific anti-Leishmania antibodies. Leishmania infantum was molecularly diagnosed in 5.4% (n = 5) of the examined animals (3/71 European rabbits, 2/7 red foxes). In many of the qPCR positive animals (4/5), the parasite DNA was more prevalent in visceral than cutaneous tissues. None of the positive animal showed signs of disease and/or macroscopic alterations of organs; low parasitic burden in all positive tissue samples was also recorded. Only one rabbit serum (i.e., 2.5%) tested positive for anti-Leishmania antibodies. The seropositive rabbit was in good health status and no amastigotes were observed in lymph-node aspirate and blood smears. This study provides first evidence of L. infantum infection in wild animals from Sicily (southern Italy). Despite the low prevalence of infection here reported, the circulation of the Leishmania in wild reservoirs in Sicily remains worthy of future investigations for a better understanding of their role in the epidemiology of the disease as well as to fine-tune control strategies in the area., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The anterior anchoring of the malleus of 30 extant species of Lagomorpha (rabbits, hares, pikas) has been studied on the basis of histological serial sections and µCT-scans. It is shown that former studies of Oryctolagus, Lepus, and Ochotona are incomplete, because the rostral part of the processus anterior of the malleus is always lacking due to damage of this extremely delicate structure. Our study shows that in perinatal stages of Leporidae the praearticulare develops a prominent processus internus that fits into a groove at the ventral side of the tegmen tympani; this "tongue and groove"-arrangement may act as a hinge. In adult stages, the rostral end of the praearticulare fuses synostotically with the medial process of the ectotympanic. Torsional strain produced by rotation around the axis of the middle ear ossicles at sound transmission must, therefore, be experienced by the extremely thin but highly elastic bony pedicle of the processus internus praearticularis. The free ending processus anterior of a late fetal Ochotona shows a short processus internus praearticularis, which does not articulate with the tegmen tympani. During postnatal development the middle ear of Ochotona becomes considerably remodelled: not only does excessive pneumatization of the tegmen tympani and tympanic cavity wall occur, but the short processus anterior is fused synostotically to a bone trabecula of the tegmen tympani meshwork. The thin and elastic bone bridges are not equivalent in Leporidae and Ochotonidae, that is, they must have evolved convergently. Fleischer's classification with Oryctolagus possessing a "freely mobile type" of middle ear ossicles cannot be supported by our observations. The same holds true for Ochotona, which does not represent a "freely mobile type" either. Thus, we suggest for the lagomorph middle ear ossicles a new category: the "bone elasticity type.", (© 2017 Wiley Periodicals, Inc.)

The skull of leporids (rabbits and hares) is highly transformed, typified by pronounced arching of the dorsal skull and ventral flexion of the facial region (i.e., facial tilt). Previous studies show that locomotor behavior influences aspects of cranial shape in leporids, and here we use an extensive 3D geometric morphometrics dataset to further explore what influences leporid cranial diversity. Facial tilt angle, a trait that strongly correlates with locomotor mode, significantly predicts the cranial shape variation captured by the primary axis of cranial shape space, and describes a small proportion (13.2%) of overall cranial shape variation in the clade. However, locomotor mode does not correlate with overall cranial shape variation in the clade, because there are two district morphologies of generalist species, and saltators and cursorial species have similar morphologies. Cranial shape changes due to phyletic size change (evolutionary allometry) also describes a small proportion (12.5%) of cranial shape variation in the clade, but this is largely driven by the smallest living leporid, the pygmy rabbit (Brachylagus idahoensis). By integrating phylogenetic history with our geometric morphometric data, we show that the leporid cranium exhibits weak phylogenetic signal and substantial homoplasy. Though these results make it difficult to reconstruct what the 'ancestral' leporid skull looked like, the fossil records suggest that dorsal arching and facial tilt could have occurred before the origin of the crown group. Lastly, our study highlights the diversity of cranial variation in crown leporids, and highlights a need for additional phylogenetic work that includes stem (fossil) leporids and includes morphological data that captures the transformed morphology of rabbits and hares., Competing Interests: Brian Kraatz is an Academic Editor for PeerJ.

The mammalian order Lagomorpha has been the subject of many morphometric studies aimed at understanding the relationship between form and function as it relates to locomotion, primarily in postcranial morphology. The leporid cranial skeleton, however, may also reveal information about their ecology, particularly locomotion and vision. Here we investigate the relationship between cranial shape and the degree of facial tilt with locomotion (cursoriality, saltation, and burrowing) within crown leporids. Our results suggest that facial tilt is more pronounced in cursors and saltators compared to generalists, and that increasing facial tilt may be driven by a need for expanded visual fields. Our phylogenetically informed analyses indicate that burrowing behavior, facial tilt, and locomotor behavior do not predict cranial shape. However, we find that variables such as bullae size, size of the splenius capitus fossa, and overall rostral dimensions are important components for understanding the cranial variation in leporids.