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Surveillance of wildlife virus impacts can be passive or active. Both approaches have their strengths and weaknesses, especially regarding cost and knowledge that can be gained. Monitoring of rabbit haemorrhagic disease virus (GI.1 and GI.2) in South Australia has utilised both strategies and their methods and gained insights are discussed. Active strategies to monitor the continuing impact of rabbit haemorrhagic disease virus 2 (GI.2) on susceptible lagomorphs in countries such as the USA, Mexico, South Africa, Spain, France and Portugal are encouraged to gain critical insights into the evolution, spread and impact of this virus. Furthermore, there are lessons here for the international monitoring of diseases in wildlife, particularly where there is a risk of them becoming zoonotic. [ABSTRACT FROM AUTHOR]

The document "New Insights into Rabbit Viral Diseases" explores the impact of viruses on rabbit populations, focusing on myxoma virus (MYXV) and rabbit hemorrhagic disease virus (RHDV). It discusses the emergence of new genotypes of RHDV with expanded host ranges, affecting various hare species and American leporids. The document also highlights the role of rabbits in different ecosystems, as both keystone species and destructive pests, and the use of biocontrol methods like RHDV in managing rabbit populations. Additionally, it emphasizes the importance of citizen science in studying lagoviruses and developing effective control strategies. [Extracted from the article]

The European domestic rabbit (Oryctolagus cuniculus domesticus) is commonly kept as a pet, with increasing popularity among pet owners. Despite the increasing body of information on lagomorph medicine and pathology, comprehensive published compilations of causes of mortality in pet rabbits are limited. We analyzed health disorders, pathology findings, and most probable causes of death in 100 pet rabbits submitted to the Anatomopathological Diagnostic Service of the Veterinary School of the University of Las Palmas de Gran Canaria, Canary Islands, Spain, from 2011 to 2022. We reviewed clinical data and gross and histopathologic findings to classify the various disorders into infectious (n = 39), noninfectious (n = 45), and idiopathic conditions (n = 16). Within infectious disease processes, Encephalitozoon cuniculi infection (n = 14) and rabbit hemorrhagic viral disease (n = 14) had the highest prevalence. Regarding the noninfectious conditions, perioperative mortality (n = 10), gastrointestinal syndrome (n = 7), and trauma (n = 6) were the most common clinicopathologic entities observed. [ABSTRACT FROM AUTHOR]

Domestic rabbits (Oryctolagus cuniculus) are commonly kept as pets or bred for laboratory investigation, meat, fur production, or a combination of these purposes. We conducted a retrospective study to assess the prevalence of diseases in domestic rabbits according to purpose. We retrieved results of autopsies, biopsies, and cytologies from 2,583 cases received at 4 diagnostic laboratories in California from 2013–2022. Rabbits were classified as pets (2,241; 86.8%), laboratory animals (92; 3.6%), meat-production animals (60; 2.3%), or multipurpose animals (190; 7.4%). A final diagnosis was reached in 2,360 (91.4%) cases and was classified by system, etiology, and type of process. Pet rabbits had the highest median age (5.9 y; vs. 3 y, 0.67 y, and 0.25 y in meat, multipurpose, and laboratory rabbits, respectively), and most of the neoplasms were diagnosed in this group (872 of the total 896 neoplasms in the study; 97.3%), with tumors of the skin, female reproductive tract, and hematolymphoid system being the most common. Laboratory rabbits had a high prevalence of infectious enterotyphlocolitis (40 of 92; 43.4%), and ~45% of those cases were due to opportunistic colibacillosis. Infectious and parasitic pneumonias were common in meat rabbits (18 of 60; 30%); pneumonic pasteurellosis accounted for >60% of those cases. Infectious cholangiohepatitides were common in multipurpose rabbits (61 of 110; 55.5%), with rabbit hemorrhagic disease representing the most common etiology (82.4% of those cases). Our results demonstrate that purpose of use can predict prevalence of disease in rabbits submitted to diagnostic laboratories. [ABSTRACT FROM AUTHOR]

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

A new form of the rabbit haemorrhagic disease virus, RHDV2, first observed in European rabbits, has spread widely among different species of hares in Europe, jackrabbits and cottontails in North America, and hares in southern Africa. However, only limited surveillance studies have been undertaken so far. It is suggested that methods developed for controlling the disease in farmed rabbits in Europe and studying the efficacy of RHDV as a biological control agent in Australia could facilitate epidemiological research on those recently affected lagomorph species. This would enable the assessment of the risk of RHDV2 to native lagomorphs, including endangered species, and the determination of the main host species of RHDV2. Because RHDV2 has not spread equally through all lagomorph species, epidemiological studies could give insights into factors important for determining host susceptibility. [ABSTRACT FROM AUTHOR]

Neuronal ceroid lipofuscinoses (NCL) are among the most prevalent neurodegenerative disorders of early life in humans. Disease‐causing variants have been described for 13 different NCL genes. In this study, a refined pathological characterization of a female rabbit with progressive neurological signs reminiscent of NCL was performed. Cytoplasmic pigment present in neurons was weakly positive with Sudan black B and autofluorescent. Immunohistology revealed astrogliosis, microgliosis and axonal degeneration. During the subsequent genetic investigation, the genome of the affected rabbit was sequenced and examined for private variants in NCL candidate genes. The analysis revealed a homozygous ~10.7 kb genomic duplication on chromosome 15 comprising parts of the MFSD8 gene, NC_013683.1:g.103,727,963_103,738,667dup. The duplication harbors two internal protein coding exons and is predicted to introduce a premature stop codon into the transcript, truncating ~50% of the wild‐type MFSD8 open reading frame encoding the major facilitator superfamily domain containing protein 8, XP_002717309.2:p.(Glu235Leufs*23). Biallelic loss‐of‐function variants in MFSD8 have been described to cause NCL7 in human patients, dogs and a single cat. The available clinical and pathological data, together with current knowledge about MFSD8 variants and their functional impact in other species, point to the MFSD8 duplication as a likely causative defect for the observed phenotype in the affected rabbit. [ABSTRACT FROM AUTHOR]

Simple Summary: Mammary carcinomas have been diagnosed with increasing frequency in pet rabbits. Prognostic biomarkers are limited, and the only available treatment is surgical excision. Additional treatment options are needed, e.g., for animals in which metastases to internal organs preclude complete tumor removal. Human breast cancer may express the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1), that represents a prognostic biomarker and a possible therapeutic target. Since previous studies revealed similarities between human breast cancer and pet rabbit mammary carcinomas, this study investigated IDO1 immunostaining in 96 mammary carcinomas of 96 pet rabbits with an average age of 5.5 years. All rabbits with reported sex were female. Variable percentages of IDO1-positive tumor cells were detected in 90 (94%) carcinomas. Furthermore, IDO1 immunostaining was observed in the secretory epithelial cells of the adjacent non-neoplastic mammary tissue. This study provides further information on the molecular features of mammary carcinomas in rabbits. It also shows similarities in IDO1 expression between rabbit mammary carcinomas and human breast cancer. These findings can have a mutual benefit. They could lead the development of novel treatment options for rabbits with mammary carcinomas. In addition, they further support the value of rabbits with mammary carcinomas for breast cancer research. For mammary carcinomas in pet rabbits, prognostic biomarkers are poorly defined, and treatment is limited to surgical excision. Additional treatment options are needed for rabbit patients for which surgery is not a suitable option. In human breast cancer, the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1) represents a prognostic biomarker and possible therapeutic target. This retrospective immunohistochemical study examined IDO1 in 96 pet rabbit mammary carcinomas with known mitotic count, hormone receptor status, and percentage of stromal tumor infiltrating lymphocytes (TILs). Tumors were obtained from 96 pet rabbits with an average of 5.5 years. All rabbits with reported sex (n = 88) were female or female-spayed. Of the carcinomas, 94% expressed IDO1, and 86% had sparse TILs consistent with cold tumors. Statistically significant correlations existed between a higher percentage of IDO1-positive tumor cells, lower mitotic counts, and increased estrogen receptor expression. The threshold for significance was IDO1 staining in >10% of tumor cells. These results lead to the assumption that IDO1 expression contributes to tumorigenesis and may represent a prognostic biomarker and possible therapeutic target also in pet rabbit mammary carcinomas. They also support the value of rabbits for breast cancer research. [ABSTRACT FROM AUTHOR]

Integrated pest management (IPM) is widely recommended for managing invasive pests to maximise effectiveness. However, the extent to which IPM occurs is typically unknown. We used the rabbit in Australia as a case study to investigate whether land managers apply IPM. Using 7415 control records voluntarily submitted to RabbitScan by land managers, we found evidence of IPM in only 39%. The extent to which integration occurred depended on the control method applied [warren ripping, poison baiting, release of rabbit haemorrhagic disease virus (RHDV), warren fumigation]. Two of four major rabbit control methods were frequently integrated with at least one other method; 73% of warren ripping and 55% of poison baiting records were associated with another different control method. In contrast, only 15% of RHDV releases and 35% of warren fumigations were integrated with another different control method. This confirmed reports that land managers view RHDV release as a 'silver bullet' and release it to avoid applying more expensive but more effective control methods. We identified control sequences that were neither biologically appropriate nor cost-efficient. In some situations, where there is a risk to other animals, or land managers lack suitable equipment, it may be difficult to apply IPM in an optimal sequence or at the optimal time. However, a greater level of control and at a proportionally lower cost could result if IPM principals were more rigorously applied, for instance, by focusing on strategic application of control methods in sequences known to be highly effective and cost-efficient. [ABSTRACT FROM AUTHOR]

Domestic rabbits (Oryctolagus cuniculus) are the fourth most common species admitted to the British Columbia Society for the Prevention of Cruelty to Animals (BC SPCA) shelter system. However, shelter data analysis has largely focused on cats and dogs and little is known about the population dynamics of rabbits in shelters. We analyzed five years of rabbit records (n = 1567) at the BC SPCA to identify trends in intake and predictors of length of stay (LOS) of rabbits. The majority of rabbits were surrendered by their owners (40.2%), with most rabbits being surrendered for human-related reasons (96.9%). Overall, rabbit intakes decreased over the study period. When analyzing by month of intake, rabbit intakes were found to be the highest in May. Most rabbits in our data were adults (46.7%), non-brachycephalic (66.7%), erect-eared (82.5%), short-furred (76.2%), and subsequently adopted (80.3%). The median LOS of rabbits was 29 days, highlighting the pressing need to improve their time to adoption. A linear model was constructed to identify predictors of LOS of adopted rabbits (n = 1203) and revealed that intake year, intake month, source of intake, age, cephalic type, and breed size significantly predicted time to adoption for rabbits (F(37, 1165) = 7.95, p < 2.2e-16, adjusted R2 = 0.18). These findings help characterize shelter population dynamics for rabbits, shed light on the challenges associated with unwanted rabbits, and offer a foundation for animal shelters to design programs and marketing strategies tailored to reduce LOS of rabbits with particular characteristics. Shelter rabbits represent an understudied population and our study highlights the importance of further research in companion rabbits. [ABSTRACT FROM AUTHOR]

Simple Summary: Domestic rabbits are one of the most commonly kept pets with a hypselodontic type of dentition, possessing teeth that grow throughout the animal's life. These animals often show signs of disease in the stomatognathic system. Dental disease can also affect the function of other organs and systems. The most common clinical signs of dental disease in domestic rabbits are the following: lack of appetite, weight loss, apathy, and difficulty chewing and swallowing food. These clinical signs are non-specific, and, therefore, a definitive diagnosis usually requires additional methods, such as an X-ray examination, a CT scan, MRI, and endoscopic examination. The most common dental disease found in this study's animals was malocclusion secondary to abnormal clinical crown abrasion and abnormal tooth growth. In domestic rabbits, osteomyelitis is a common complication of dental abscesses. Computed tomography is an invaluable diagnostic method in the diagnosis of dental disease and secondary diseases of the head area, such as inflammation of the nasal cavities or otitis media, in pet rabbits. Domestic rabbits have teeth that grow throughout the animal's life and are prone to disease. Clinical signs of dental disease in domestic rabbits are non-specific, and, therefore, a definitive diagnosis usually requires additional methods. This study was carried out on a group of 105 domestic rabbits aged 3 to 9 years. In total, 90 domestic rabbits with dental disease visible on CT images and other secondary diseases of the head area qualified for this study. Malocclusion was found in 57 (63.3%). Retrograde elongation of the tooth apices in the mandible was present in 39 (43.3%), and it was present in the maxilla in 48 (53%). Clinical tooth crowns were overgrown in 39 (43%). Dental abscesses were present in 54 (63%). Secondary to the presence of a dental abscess, osteomyelitis was found in 43 (79% of the animals with a dental abscess). Dental inflammatory resorption was found in 36 (40%). Secondary to dental disease, nasal cavity inflammation was found in 18 (20%). Otitis media was present in six (6.7%). The most common dental disease found in this study's animals was malocclusion secondary to abnormal clinical crown abrasion and abnormal tooth growth. In domestic rabbits, osteomyelitis is a common complication of dental abscesses. Computed tomography is an invaluable diagnostic method in the diagnosis of dental disease and secondary diseases of the head area, such as inflammation of the nasal cavities or otitis media, in pet rabbits. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhage disease virus 2 (RHDV2) is a highly pathogenic lagovirus that causes lethal disease in rabbits and hares (lagomorphs). Since its first detection in Europe in 2010, RHDV2 has spread worldwide and has been detected in over 35 countries so far. Here, we provide the first detailed report of the detection and subsequent circulation of RHDV2 in New Zealand. RHDV2 was first detected in New Zealand in 2018, with positive samples retrospectively identified in December 2017. Subsequent time-resolved phylogenetic analysis suggested a single introduction into the North Island between March and November 2016. Genetic analysis identified a GI.3P-GI.2 variant supporting a non-Australian origin for the incursion; however, more accurate identification of the source of the incursion remains challenging due to the wide global distribution of the GI.3P-GI.2 variant. Furthermore, our analysis suggests the spread of the virus between the North and South Islands of New Zealand at least twice, dated to mid-2017 and around 2018. Further phylogenetic analysis also revealed a strong phylogeographic pattern. So far, no recombination events with endemic benign New Zealand rabbit caliciviruses have been identified. This study highlights the need for further research and surveillance to monitor the distribution and diversity of lagoviruses in New Zealand and to detect incursions of novel variants. [ABSTRACT FROM AUTHOR]

OBJECTIVE: This study aimed to characterize the bacterial and eukaryotic microbiota of the gastrointestinal (GI) tract in domestic rabbits and to evaluate the effect of different diet characteristics, such as pelleting, extrusion, and hay supplementation. ANIMALS: 30 New Zealand White rabbits (15 male and 15 female; 6 to 7 months old) were fed 1 of 6 diets (5 rabbits per diet) for 30 days after an initial acclimation period. At the end of the trial, samples were collected from the stomach, small intestine, cecum, large intestine, and hard feces. METHODS: The samples were analyzed using 16S rRNA and internal transcribed spacer 1 region-targeted amplicon sequencing. RESULTS: The bacterial microbiota was distinct between the foregut and hindgut. The most abundant bacterial genera included an unclassified genus in the Bacteroidales order and Alistipes. Candida was the most abundant genus in the eukaryotic dataset. In the bacterial dataset, diet No Hay/Pellet E was shown to have lower diversity (Shannon diversity, P < .05) compared to all diet groups except for No Hay/Pellet M. Few significant differences in alpha-diversity indexes between diet groups were detected in the eukaryotic dataset. CLINICAL RELEVANCE: Our findings demonstrated that feeding hay had a significant effect on the beta diversity of the bacterial microbiota. Given the prevalence of gastrointestinal disease in the domestic rabbit population, furthering our understanding of what constitutes a healthy rabbit microbiota and the effects of different diets on the microbial community can help veterinarians implement better intervention strategies and allow pet owners to provide the best level of care. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRs) are a group of small, 17–25 nucleotide, non-coding RNA that regulate gene expression at the post-transcriptional level. To date, little is known about the molecular signatures of regulatory interactions between miRs and apoptosis and oxidative stress in viral diseases. Lagovirus europaeus is a virus that causes severe disease in rabbits (Oryctolagus cuniculus) called Rabbit Hemorrhagic Disease (RHD) and belongs to the Caliciviridae family, Lagovirus genus. Within Lagovirus europaeus associated with RHD, two genotypes (GI.1 and GI.2) have been distinguished, and the GI.1 genotype includes four variants (GI.1a, GI.1b, GI.1c, and GI.1d). The study aimed to assess the expression of miRs and their target genes involved in apoptosis and oxidative stress, as well as their potential impact on the pathways during Lagovirus europaeus—two genotypes (GI.1 and GI.2) infection of different virulences in four tissues (liver, lung, kidneys, and spleen). The expression of miRs and target genes related to apoptosis and oxidative stress was determined using quantitative real-time PCR (qPCR). In this study, we evaluated the expression of miR-21 (PTEN, PDCD4), miR-16b (Bcl-2, CXCL10), miR-34a (p53, SIRT1), and miRs—related to oxidative stress—miR122 (Bach1) and miR-132 (Nfr-2). We also examined the biomarkers of both processes (Bax, Bax/Bcl-2 ratio, Caspase-3, PARP) and HO-I as biomarkers of oxidative stress. Our report is the first to present the regulatory effects of miRs on apoptosis and oxidative stress genes in rabbit infection with Lagovirus europaeus—two genotypes (GI.1 and GI.2) in four tissues (liver, lungs, kidneys, and spleen). The regulatory effect of miRs indicates that, on the one hand, miRs can intensify apoptosis (miR-16b, miR-34a) in the examined organs in response to a viral stimulus and, on the other hand, inhibit (miR-21), which in both cases may be a determinant of the pathogenesis of RHD and tissue damage. Biomarkers of the Bax and Bax/Bcl-2 ratio promote more intense apoptosis after infection with the Lagovirus europaeus GI.2 genotype. Our findings demonstrate that miR-122 and miR-132 regulate oxidative stress in the pathogenesis of RHD, which is associated with tissue damage. The HO-1 biomarker in the course of rabbit hemorrhagic disease indicates oxidative tissue damage. Our findings show that miR-21, miR-16b, and miR-34a regulate three apoptosis pathways. Meanwhile, miR-122 and miR-132 are involved in two oxidative stress pathways. [ABSTRACT FROM AUTHOR]

Several conventional and recently available tools are available for an integrated control of European rabbits in Australia. We quantified the impact of the release of rabbit haemorrhagic disease virus K5 (RHDV K5, hereafter K5) and pindone (2‐pivalyl‐1,3‐indandione) baiting at 13 sites within Cudlee Creek fire scar in the Adelaide Hills, South Australia. K5 release was followed by pindone baiting between December 2021 and March 2022; the application of both control methods followed industry best practice. We counted rabbits using spotlights before and after the application of both control methods. Fly samples and livers from dead rabbits were collected to track K5 transmission within and between sites, and to detect the natural circulation of rabbit haemorrhagic disease virus 2 (RHDV2). K5 release had minimal impact on rabbit populations, with treated populations increasing by a mean of 65.5% at 14 days post‐release and 27.9% at 77 days post‐K5 release across all sites, comparable to the changes at control sites. K5 detection in flies up to 77 days post its release, and its detection in rabbit livers, demonstrates that it can survive and transmit in the environment for prolonged periods and that it can lethally infect some rabbits. This limited impact of K5 is consistent with previous studies and may be explained by pre‐existing RHDV/RHDV2 immunity in the target populations or the presence of young rabbits with natural innate RHDV immunity. The detection of K5 in flies from control sites demonstrates that it was vectored beyond its release location. A reduction in rabbit counts post‐pindone baiting was observed at most treatment sites, with a mean population reduction of 36.6% across all sites. Landholders need to carefully and strategically plan their integrated rabbit control programmes. Not all combinations of controls, even if theoretically logical, achieve meaningful outcomes for rabbit management. [ABSTRACT FROM AUTHOR]

Pathogenic lagoviruses (Rabbit hemorrhagic disease virus, RHDV) are widely spread across the world and are used in Australia and New Zealand to control populations of feral European rabbits. The spread of the non-pathogenic lagoviruses, e.g., rabbit calicivirus (RCV), is less well studied as the infection results in no clinical signs. Nonetheless, RCV has important implications for the spread of RHDV and rabbit biocontrol as it can provide varying levels of cross-protection against fatal infection with pathogenic lagoviruses. In Chile, where European rabbits are also an introduced species, myxoma virus was used for localised biocontrol of rabbits in the 1950s. To date, there have been no studies investigating the presence of lagoviruses in the Chilean feral rabbit population. In this study, liver and duodenum rabbit samples from central Chile were tested for the presence of lagoviruses and positive samples were subject to whole RNA sequencing and subsequent data analysis. Phylogenetic analysis revealed a novel RCV variant in duodenal samples that likely originated from European RCVs. Sequencing analysis also detected the presence of a rabbit astrovirus in one of the lagovirus-positive samples. [ABSTRACT FROM AUTHOR]

Background: Rabbits are extremely sensitive to pain. There is evidence on whether non‐steroidal anti‐inflammatory medication should be administered before or after surgery in rabbits. Objectives: In this study, we compared the pain scores and behaviours of rabbits administered meloxicam before or after routine castration. Methods: Forty‐two healthy rabbits undergoing castration were randomly assigned to receive meloxicam (1 mg/kg SC) 1 h before or upon recovery from anaesthesia. Several parameters were recorded, including the occurrence of eating, defecating and pain scores postoperatively and change in heart rate intraoperatively. Results: At 4 h after surgery, eating was observed in 43% (9/21) of rabbits that received meloxicam before neutering and in 29% (6/21) of rabbits that received meloxicam after. The morning after the surgery, eating was observed in 62% (13/21) of rabbits that received meloxicam before neutering and in 57% (12/21) of rabbits that received meloxicam after. The odds of not eating increased by 1.16 times (1.03–1.29; p = 0.01) per month of age, whereas there was no significant association between eating and administration of meloxicam before or after surgery, body weight and time. Similarly, defecation, pain score and change in heart rate did not differ in animals receiving meloxicam before or after surgery. Conclusions: Administration of meloxicam in rabbits undergoing castration before or after surgery does not have a relevant effect on eating, defecating or pain score. Younger rabbits show less obvious signs of pain and are more likely to resume eating habits sooner after surgery compared to older rabbits. [ABSTRACT FROM AUTHOR]

Digestive problems, both those with a clear pathogenic origin (e.g., Escherichia coli) and those without obvious pathogen involvement [e.g., syndromes like epizootic rabbit enteropathy (ERE)], are common in production rabbits and account for the majority of losses in meat rabbit production. A multitude of nutritional, genetic and housing factors have been found to play a role in the occurrence of digestive problems. However, the exact early pathophysiological mechanism, including the links between aforementioned risk factors and subsequent development and expression of gastrointestinal disease, is less clear, especially in non-specific enteropathies without obvious pathogen involvement. In this review, we aim to shed more light on the derailment of the normal gastrointestinal functioning in rabbits. We discuss a conceptual integrated view of this derailment, based on an "overload" pathway and a "chymus jam" pathway, which may occur simultaneously and interact. The "overload" pathway centers around exposure to excess amounts of easily fermentable substrate (e.g., starch and protein) that might be incompletely digested prior to entering the caecum. Once there, hyperfermentationmay result in changes in caecal pH and inhibition of the normal microflora. The second pathway centers around a chymus jam resulting froma compromised passage rate. Here, reduced hindgutmotility (e.g., resulting from stress or limited fiber supply) leads to reduced flow of digesta and increased caecal retention times, which might lead to the production of abnormal caecal fermentation products and subsequent inhibition of the normal microflora. A central role in the presumed mechanism is attributed to the fusus coli. We discuss the suggested mechanisms behind both pathways, as well as the empirical substantiation and alignment between theoretical concepts and observations in practice. The proposed hypotheses may explain the effect of time-based restriction to prevent ERE, which is widely applied in practice but to date not really understood, and suggest that the particle size of fibermay be a key point in the normal functioning of the colon and fusus coli. Further insight into the circumstances leading to the derailment of physiological processes in the rabbit hindgut could provide a meaningful starting point to help improve their gastrointestinal resilience. [ABSTRACT FROM AUTHOR]

Rabbit Fibroma is a Leporipoxviral disease and is considered the third most common cutaneous neoplasm in pet rabbits. Two domestic rabbits (Oryctolagus cuniculus) were submitted to the veterinary clinic due to the presence of a nodule on the lip. Histologically, epithelial cells of the epidermis and hair follicles showed mild to moderate ballooning degeneration, spongiosis, and several eosinophilic intracytoplasmic inclusion bodies. The dermis was expanded by atypical spindle cells that also showed eosinophilic intracytoplasmic inclusion bodies. The tissues were evaluated by using transmission electron microscopy. In both cases, keratinocytes exhibit several electron dense and pleomorphic intracytoplasmic viral particles consistent with Poxviruses. To our knowledge, this is the first case report of Rabbit Fibroma Virus infection in Domestic Rabbits in Mexico. [ABSTRACT FROM AUTHOR]

Following the recent domestication process of the European rabbit (Oryctolagus cuniculus), many different breeds and lines, distinguished primarily by exterior traits such as coat colour, fur structure and body size and shape, have been constituted. In this study, we genotyped, with a high‐density single‐nucleotide polymorphism panel, a total of 645 rabbits from 10 fancy breeds (Belgian Hare, Champagne d'Argent, Checkered Giant, Coloured Dwarf, Dwarf Lop, Ermine, Giant Grey, Giant White, Rex and Rhinelander) and three meat breeds (Italian White, Italian Spotted and Italian Silver). ADMIXTURE analysis indicated that breeds with similar phenotypic traits (e.g. coat colour and body size) shared common ancestries. Signatures of selection using two haplotype‐based approaches (iHS and XP‐EHH), combined with the results obtained with other methods previously reported that we applied to the same breeds, we identified a total of 5079 independent genomic regions with some signatures of selection, covering about 1777 Mb of the rabbit genome. These regions consistently encompassed many genes involved in pigmentation processes (ASIP, EDNRA, EDNRB, KIT, KITLG, MITF, OCA2, TYR and TYRP1), coat structure (LIPH) and body size, including two major genes (LCORL and HMGA2) among many others. This study revealed novel genomic regions under signatures of selection and further demonstrated that population structures and signatures of selection, left into the genome of these rabbit breeds, may contribute to understanding the genetic events that led to their constitution and the complex genetic mechanisms determining the broad phenotypic variability present in these untapped rabbit genetic resources. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus (RHDV) is established as a landscape-scale biocontrol that assists the management of invasive European rabbits and their impacts in both Australia and New Zealand. In addition to this, it is also available to land managers to augment rabbit control efforts at a local scale. However, current methods of deploying RHDV to rabbits that rely on the consumption of virus-treated baits can be problematic as rabbits are reluctant to consume bait when there is abundant, green, protein-rich feed available. We ran a suite of interrupted time-series experiments to compare the duration of infectivity of two conventional (carrot and oat baits) and two novel (meat bait and soil burrow spray) methods of deploying RHDV to rabbits. All methods effectively killed exposed rabbits. Soil burrow spray and carrot baits resulted in infection and mortality out to 5 days post their deployment in the field, and meat baits caused infection out to 10 days post their deployment. In contrast, oat baits continued to infect and kill exposed rabbits out to 20 days post deployment. Molecular assays demonstrated high viral loads in deployed baits beyond the duration for which they were infectious or lethal to rabbits. Based on our results, we suggest that the drying of meat baits may create a barrier to effective transmission of RHDV by adult flies within 10 days. We therefore hypothesise that fly larvae production and development on infected tissues is critical to prolonged viral transmission from meat baits, and similarly from carcasses of RHDV mortalities, via mechanical fly vectors. Our study demonstrates that meat baits and soil spray could provide additional virus deployment options that remove the need for rabbits to consume baits at times when they are reluctant to do so. [ABSTRACT FROM AUTHOR]

This article discusses the therapeutic management of notoedric mange in rabbits. The study was conducted in India and involved two rabbits with alopecia, rough hair coat, and intense pruritus. The rabbits were treated with injections of Ivermectin and Chlorpheniramine maleate, as well as topical application of Povidone-iodine. The treatment was successful, with the rabbits showing improvement and complete recovery within 15 days. Notoedric mange is a highly contagious disease caused by burrowing mites, and it can spread rapidly through direct contact or interaction with the environment. Ivermectin is an effective treatment for mange in rabbits. [Extracted from the article]

The Journal of Veterinary Diagnostic Investigation has published a special issue on lagomorph disease investigations. Lagomorphs are mammals in the order Lagomorpha, which includes pikas, hares, jackrabbits, and various species of rabbits. The issue includes articles on lagomorph pathology and laboratory testing, aimed at helping veterinary laboratory diagnosticians, pathologists, and other veterinarians. The interest in lagomorph science has grown in recent years, with an increasing number of pet rabbit owners and a rising interest in diseases that affect rabbits. The issue also covers diseases and pathology of native North American lagomorphs, the emergence and spread of rabbit hemorrhagic disease virus, and other topics related to lagomorph health. [Extracted from the article]

Background: Immunoglobulin A (IgA) is the mammalian mucosal antibody, providing an important line of defense against pathogens. With 15 IgA subclasses, the European rabbit has an extremely complex IgA system, strikingly more complex than most other mammals, which have only one IgA or, in the case of hominoids, two IgA subclasses. Similar to the two hominoid primate IGHA genes, the expansion of the rabbit IGHA genes appears to have begun in an ancestral lagomorph since multiple IgA copies were found by Southern blot analysis for the genera Sylvilagus, Lepus, and Ochotona. Results: To gain a better insight into the extraordinary lagomorph IgA evolution, we sequenced, for the first time, expressed IgA genes for two Lepus species, L. europaeus and L. granatensis. These were aligned with the 15 rabbit IgA isotypes, and evolutionary analyses were conducted. The obtained phylogenetic tree shows that the Lepus IgA sequences cluster with and among the rabbit IgA isotypes, and the interspecies and intraspecies nucleotide genetic distances are similar. A comparison of the amino acid sequences of the Lepus and rabbit IgA confirms that there are two trans-species polymorphisms and that the rabbit and Lepus sequences share a common genetic pool. In fact, the main differences between the studied leporids IgAs reside in the characteristics of the hinge region. Conclusion: The Lepus IgA sequences we have obtained strongly suggest that the great expansion of the leporid IGHA genes occurred in a common ancestral species and was then maintained in the descendants. A strong selective pressure caused the extraordinary expansion of the IGHA genes but then subsided, leading to the maintenance of the acquired polymorphisms in the descendants, with little subsequent divergence. This is a unique evolutionary pattern in which an ancient gene expansion has been maintained for approximately 18 million years. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The European rabbit (Oryctolagus cuniculus) populations of the Iberian Peninsula have been severely affected by the emergence of the rabbit haemorrhagic disease virus (RHDV) Lagovirus europaeus/GI.2 (RHDV2/b). Bushflies and blowflies (Muscidae and Calliphoridae families, respectively) are important RHDV vectors in Oceania, but their epidemiological role is unknown in the native range of the European rabbit. In this study, scavenging flies were collected between June 2018 and February 2019 in baited traps at one site in southern Portugal, alongside a longitudinal capture-mark-recapture study of a wild European rabbit population, aiming to provide evidence of mechanical transmission of GI.2 by flies. Fly abundance, particularly from Calliphoridae and Muscidae families, peaked in October 2018 and in February 2019. By employing molecular tools, we were able to detect the presence of GI.2 in flies belonging to the families Calliphoridae, Muscidae, Fanniidae and Drosophilidae. The positive samples were detected during an RHD outbreak and absent in samples collected when no evidence of viral circulation in the local rabbit population was found. We were able to sequence a short viral genomic fragment, confirming its identity as RHDV GI.2. The results suggest that scavenging flies may act as mechanical vectors of GI.2 in the native range of the southwestern Iberian subspecies O. cuniculus algirus. Future studies should better assess their potential in the epidemiology of RHD and as a tool for monitoring viral circulation in the field. [ABSTRACT FROM AUTHOR]

Simple Summary: The emergence of two strains of rabbit haemorrhagic disease (i.e., GI.1 and GI.2) in the1990s and 2010s, respectively, has been the primary determinant for the decline of wild European rabbits within their native range. We compared the impact of both strains on the wild rabbit populations in Spain using national hunting bags. Our findings showed that GI.1 had a greater impact on wild rabbit populations than GI.2. This disparity is likely to be explained by several factors, such as climatic conditions, host resistance improvement, virulence attenuation, and population density, among others. The outbreaks of two strains of rabbit haemorrhagic disease (RHD) (GI.1 and GI.2) in the Iberian Peninsula have caused substantial economic losses in commercial rabbitries and have affected the conservation of rabbit-sensitive predators due to the dramatic decline of their natural populations. However, the assessment of the impact of both RHD strains on wild rabbit populations has been limited to a few small-scale studies. Little is known about the overall impact within its native range. In this study, we described and compared the effects of GI.1 and GI.2 countrywide by using time series of hunting bag data widely available across the country and compared their trend during the first eight years after the first outbreak of GI.1 (i.e., 1998) and GI.2 (i.e., 2011), respectively. We used Gaussian generalised additive models (GAM) with the number of hunted rabbits as the response variable and year as the predictor to evaluate the non-linear temporal dynamics of the population at the national and regional community levels. The first GI.1 caused a population decline of around 53%, affecting most Spanish regional communities where the disease occurred. The positive trend observed after GI.1 in Spain ended with the initial outbreak of GI.2, which did not appear to cause a national population decline. In contrast, we found significant variability in the rabbit population trend among regional communities, where some increased, and others decreased. Such a disparity is unlikely to be explained by a single factor; rather, it appears to result from several factors, such as climatic conditions, host resistance improvement, virulence attenuation, or population density. Our study suggests that a national comprehensive hunting bag series could aid in elucidating the differences in the impact of emerging diseases on a large scale. Future research should focus on national longitudinal serological studies to shed light on the immunological status of rabbit populations in different regions to better understand the evolution of RHD strains and the resistance gained by the wild populations. [ABSTRACT FROM AUTHOR]

There are many advantages associated with the determination of the level of corticosterone in rabbits from saliva, since this is a non-invasive sample collection method that does not affect their welfare and provides a reliable reflection of the state of the animal at a given moment without the results being distorted as they may be, for example, when blood samples are taken. The aim of this study was to determine the diurnal rhythm in the concentration of corticosterone in the saliva of the domestic rabbit. Saliva samples were taken from six domestic rabbits five times during the daytime (at 6:00, 9:00, 12:00, 15:00 and 18:00) over the course of three consecutive days. The levels of corticosterone in the saliva of the individual rabbits displayed a diurnal rhythm during the course of the day, with a significant increase between 12:00 and 15:00 (p < 0.05). No statistically significant difference in the concentrations of corticosterone in the saliva of the individual rabbits was demonstrated. Although the basal value of corticosterone is not known in rabbits and is difficult to determine, the results of our study show the pattern of fluctuations in the concentration of corticosterone in the saliva of rabbits during the daytime. [ABSTRACT FROM AUTHOR]

Simple Summary: Vitamin D is an important hormone that can be acquired through diet or exposure to ultraviolet B (UVB) radiation. Few studies have evaluated the effects of UVB radiation on vitamin D concentrations in the domestic rabbit (Oryctolagus cuniculus); however, initial findings have found they can increase their serum 25-hydroxyvitamin D3 (25-OHD3) following 12 h of artificial UVB exposure. Current husbandry recommendations for rabbits do not include specific UVB lighting requirements. Rabbits are a common pet and research model and are frequently housed indoors without access to natural UVB lighting. Rabbits that are chronically vitamin D deficient may develop mineral deficiencies that can lead to poor calcification of the teeth and skull, predisposing these animals to dental abnormalities, bone infections, and other debilitating diseases. While initial results suggest artificial UVB is positive for rabbits, UVB can also be detrimental to the health of vertebrates. The aim of this study was to determine if shorter-duration UVB exposure could also increase 25-OHD3 concentrations. Rabbits were provided 6 h of artificial UVB daily for 14 days, and there was a significant increase in 25-OHD3 concentrations over time. These findings affirm that rabbits can use short-duration artificial UVB to increase 25-OHD3 concentrations. Vitamin D is an important hormone that can be acquired through diet, exposure to ultraviolet B (UVB) radiation, or a combination of these methods. In domestic rabbits (Oryctolagus cuniculus), both methods appear viable, but there is limited research evaluating the effects of UVB on this species. Previous studies found that 12 h of artificial UVB radiation significantly increased 25-hydroxyvitamin D3 (25-OHD3) concentrations over time. While these findings suggest UVB can be beneficial in rabbits, this form of radiation can also be detrimental to vertebrates. The purpose of this study was to determine if shorter-duration UVB could elicit a similar physiological response in rabbits while minimizing potential negative effects. Six rabbits were used for this pilot study. The baseline serum 25-OHD3 was measured for each rabbit and following 14 days of 6 h/day exposure to artificial UVB, a second 25-OHD3 sample was collected. There was a significant increase (p = 0.001) in serum 25-OHD3 over time (Baseline: 27.7 ± 8.1 nmol/L; Day 14: 79.8 ± 9 nmol/L). This study affirmed that 6 h of UVB produced 25-OHD3 concentrations similar to those found in rabbits exposed to 12 h of UVB. Future studies should continue to determine how the duration of UVB exposure affects 25-OHD3 concentrations. [ABSTRACT FROM AUTHOR]

Simple Summary: There has been an increase in domestic rabbit ownership in the recent years. The type of dentition of this exotic (non-traditional) animal pet, predisposes it to the development of Acquired Dental Disease (ADD), a disease that affects tooth quality, generates malocclusion, among other dental and non-dental consequences. The aim of this study was to assess the factors that modify the risk for ADD in domestic pet rabbits. For this, 1420 owned rabbits' clinical records from a private practice from 2018 to 2021 were used, recording several variables, including the clinical signs at diagnosis. ADD was detected in 25.4% of the individuals, mostly on their cheek teeth. In addition, age and sex (male) were found to be significant risk factors for ADD. In contrast, a free lifestyle and consuming hay in the diet were protective factors. Understanding the factors that modify the risk of ADD increases the chances of prevention and helps to improve the education of owners and/or guardians of domestic rabbits. Rabbits (Oryctolagus cuniculus) have elodont dentition, a characteristic that predisposes them to the development of Acquired Dental Disease (ADD), which is a multifactorial disease. The aim of this study was to assess the risk factors for ADD in domestic pet rabbits. To do this, a retrospective analysis of 1420 rabbits treated at a private practice during 2018–2021 was performed. For this, several variables were retrieved from clinical records, in addition to signology at the time of diagnosis. ADD was found on 25.4% of rabbits, mostly on their cheek teeth. In addition, age (OR = 1.029; 95% CI = 1.023–1.035; p < 0.001) and sex (male) (OR = 1.591; 95% CI = 1.226–2.064; p < 0.001) were found to be significant risk factors for ADD. In contrast, a free lifestyle (OR = 0.565; 95% CI = 0.362–0.882; p = 0.012) and consuming hay in the diet (OR = 0.323; 95% CI = 0.220–0.473; p < 0.001) were protective factors. In conclusion, ADD has a high prevalence and is usually underdiagnosed, highlighting the need for an exhaustive evaluation of patients during the clinical examination. This study improves our knowledge of ADD and its prevention. [ABSTRACT FROM AUTHOR]

Simple Summary: In rabbits, milk is the primary source of nutrition from early growth to weaning. The ability of the mother rabbit to produce milk, which is also influenced by the maternal genotype, is particularly important in the case of the larger litters. The hormone prolactin is responsible for the initiation and maintenance of lactation and for the synthesis of the major components of milk. Prolactin acts through membrane receptors in target tissues. Point mutations and microsatellites in receptor genes can affect production characteristics. Our aim was to examine the prolactin receptor gene in a wild rabbit (Oryctolagus cuniculus) population with a diverse genetic background. Our hypothesis was that the detected polymorphisms could be associated with milk production. By sequencing the promoter region of the PRLR gene, we detected four point mutations and one microsatellite. Among the genotypes of point mutations in the regulatory region of the PRLR gene, the homozygous genotype and the short repeat of the microsatellite resulted in higher milk production. These could be potential marker candidates for the development of marker-assisted selection. 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. [ABSTRACT FROM AUTHOR]

Since their introduction in 1859, European rabbits (Oryctolagus cuniculus) have had a devastating impact on agricultural production and biodiversity in Australia, with competition and land degradation by rabbits being one of the key threats to agricultural and biodiversity values in Australia. Biocontrol agents, with the most important being the rabbit haemorrhagic disease virus 1 (RHDV1), constitute the most important landscape-scale control strategies for rabbits in Australia. Monitoring field strain dynamics is complex and labour-intensive. Here, using phylodynamic models to analyse the available RHDV molecular data, we aimed to: investigate the epidemiology of various strains, use molecular data to date the emergence of new variants and evaluate whether different strains are outcompeting one another. We determined that the two main pathogenic lagoviruses variants in Australia (RHDV1 and RHDV2) have had similar dynamics since their release, although over different timeframes (substantially shorter for RHDV2). We also found a strong geographic difference in their activities and evidence of overall competition between the two viruses. [ABSTRACT FROM AUTHOR]

The vegetation of coastal sand dunes is characterized by high species diversity and comprises some of the rarest vegetation types in North-Western Europe. Among them are dune grassland communities whose species richness relies on grazing. Those communities are assessed as a priority habitat type under the Natura 2000 legislation. In autumn 1990, Galloway cows and Nordic Fjord horses were introduced in the coastal dunes of Meijendel near The Hague (52º 7‘N; 4º 20’E), The Netherlands, to reduce encroachment of tall grasses and shrubs, to develop bare sand patches, and as such facilitating diverse vegetation structures in the dune grasslands. In the 1950s, decades before the introduction of livestock, 41 permanent plots were installed. On average, they were examined every four years. Our study hypothesised that the livestock grazing in the set densities would halt progressive succession and facilitate regressive succession. Up to 1990, we observed an equilibrium between progressive and regressive succession. After 1990, however, our data showed a pronounced progressive succession contradicting the hypothesized effect of the livestock grazing. We relate the main observed patterns with two factors linked to rabbit populations: (i) the myxomatosis outbreak in 1954 and (ii) the rabbit Viral Haemorrhagic Disease (rVHD-1) outbreak in 1989. In addition to livestock grazing, rabbits block progressive succession by feeding on seedlings of shrub and tree species and digging burrows, creating small-scale mosaics of bare sand and initiate blowout development when collapsing. We state that the substantial decrease in rabbit numbers due to the viral diseases likely caused the observed increase of shrubs and trees in the study area's permanent plots. Climate change might have contributed to the observed increase in autonomous blowout development since 2001, as well as a decrease in atmospheric nitrogen deposition since 1990, after a strong increase the decades before. [ABSTRACT FROM AUTHOR]

A long‐term active epidemiological surveillance programme was conducted to determine seroprevalence to myxoma virus (MYXV), infection prevalence and spatiotemporal patterns and factors associated with MYXV circulation in wild rabbits (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems. A total of 2376 animals were sampled over four study periods: 2009–2012 (P1), 2012–2015 (P2), 2015–2018 (P3) and 2018–2021 (P4). Antibodies against MYXV were detected by a commercial indirect ELISA in 59.9% (1424/2376; 95% CI: 58.0–61.9) of wild rabbits. At least one seropositive animal was detected on 131 (96.3%) of 136 game estates sampled. MYXV infection was confirmed by PCR in 94 of 1063 (8.8%; 95% CI: 7.3–10.7) wild rabbits. Circulation of the novel recombinant MYXV (ha‐MYXV) was not found in wild rabbits analysed during P4. Five statistically significant spatiotemporal clusters of high MYXV seroprevalence were identified using a Bernoulli model: one in P2 and four in P3. A generalized linear mixed model (GLMM) analysis identified sampling season (autumn), age (adult and juvenile), outbreaks of myxomatosis in the month prior to sampling, mean annual temperature, humidity and seropositivity to rabbit haemorrhagic disease virus as factors potentially linked with MYXV seropositivity. GLMM analysis identified outbreaks of myxomatosis in the month prior to sampling, MYXV seropositivity and presence of lesions compatible with myxomatosis as factors associated with MYXV infection. The results indicate high exposure, widespread but non‐homogeneous distribution, and endemic circulation of MYXV in wild rabbit populations in southern Spain during the last decade. Prevalence of antibodies against MYXV showed fluctuations both within the year and over the study periods, revealing variations in the immunity of wild rabbit populations in Mediterranean ecosystems that could increase the risk of MYXV re‐emergence in immunologically naïve populations. The present study highlights the importance of long‐term surveillance to better understand the epidemiology of MYXV in wild lagomorphs. [ABSTRACT FROM AUTHOR]

We made 2 Z-based in situ hybridization (ISH) probes for the detection of rabbit hemorrhagic disease virus 2 (RHDV2; Lagovirus GI.2) nucleic acid in formalin-fixed, paraffin-embedded tissues from European rabbits (Oryctolagus cuniculus) that had died during an outbreak of RHD in Washington, USA. One probe system was made for detection of negative-sense RNA (i.e., the replicative intermediate RNA for the virus), and the other probe system was constructed for detection of genomic and mRNA of the virus (viral mRNA). Tissue sets were tested separately, and the viral mRNA probe system highlighted much broader tissue distribution than that of the replicative intermediate RNA probe system. The latter was limited to liver, lung, kidney, spleen, myocardium, and occasional endothelial staining, whereas signal for the viral mRNA was seen in many more tissues. The difference in distribution suggests that innate phagocytic activity of various cell types may cause overestimation of viral replication sites when utilizing ISH of single-stranded, positive-sense viruses. [ABSTRACT FROM AUTHOR]

Simple Summary: This 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. [ABSTRACT FROM AUTHOR]

Estimation of the diagnostic performance of serological tests often relies on another test assumed as a reference or on samples of known infection status, yet both are seldom available for emerging pathogens in wildlife. Longitudinal disease serological data can be analysed through multi‐event capture–mark–recapture (MECMR) models accounting for the uncertainty in state assignment, allowing us to estimate epidemiological parameters such as incidence and mortality. We hypothesized that by estimating the uncertainty in state assignment, MECMR models estimate the diagnostic performance of serological tests for rabbit haemorrhagic disease virus (RHDV) and myxoma virus (MYXV). We evaluated this hypothesis on longitudinal serological data of three tests of RHDV and one test of MYXV in two populations of the European rabbit (Oryctolagus cuniculus algirus). First, we selected the optimal cut‐off threshold for each test using finite mixture models, a reference method not relying on reference tests or samples. Second, we used MECMR models to compare the diagnostic sensitivity (Se) and specificity (Sp) of the three tests for RHDV. Third, we compared the estimates of diagnostic performance by MECMR and finite mixture models across a range of cut‐off values. The MECMR models showed that the RHDV test employing GI.2 antigens (Se: 100%) outperformed two tests employing GI.1 antigens (Se: 21.7% ± 8.6% and 8.7% ± 5.9%). At their selected cut‐offs (2.0 for RHDV GI.2 and 2.4 for MYXV), the estimates of Se and Sp were concordant between the MECMR and finite mixture models. Over the duration of the study (May 2018 to September 2020), the monthly survival of European rabbits seropositive for MYXV was significantly higher than that of seronegative rabbits (82.7% ± 4.9% versus 61.5% ± 12.7%) at the non‐fenced site. We conclude that MECMR models can reliably estimate the diagnostic performance of serological tests for RHDV and MYXV in European rabbits. This conclusion could extend to other diagnostic tests and host‒pathogen systems. Longitudinal disease surveillance data analysed through MECMR models allow the validation of diagnostic tests for emerging pathogens in novel host species while simultaneously estimating epidemiological parameters. [ABSTRACT FROM AUTHOR]

Australia is known for its long history of using biocontrol agents, such as myxoma virus (MYXV) and rabbit haemorrhagic disease virus (RHDV), to manage wild European rabbit populations. Interestingly, while undertaking RHDV surveillance of rabbits that were found dead, we observed that approximately 40% of samples were negative for RHDV. To investigate whether other infectious agents are responsible for killing rabbits in Australia, we subjected a subset of these RHDV‐negative liver samples to metatranscriptomic sequencing. In addition, we investigated whether the host transcriptome data could provide additional differentiation between likely infectious versus non‐infectious causes of death. We identified transcripts from several Clostridia species, Pasteurella multocida, Pseudomonas spp., and Eimeria stiedae, in liver samples of several rabbits that had died suddenly, all of which are known to infect rabbits and are capable of causing disease and mortality. In addition, we identified Hepatitis E virus and Cyniclomyces yeast in some samples, both of which are not usually associated with severe disease. In one‐third of the sequenced total liver RNAs, no infectious agent could be identified. While metatranscriptomic sequencing cannot provide definitive evidence of causation, additional host transcriptome analysis provided further insights to distinguish between pathogenic microbes and commensals or environmental contaminants. Interestingly, three samples where no pathogen could be identified showed evidence of up‐regulated host immune responses, while immune response pathways were not up‐regulated when E. stiedae, Pseudomonas, or yeast were detected. In summary, although no new putative rabbit pathogens were identified, this study provides a robust workflow for future investigations into rabbit mortality events. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus 2 (RHDV2) is now the dominant calicivirus circulating in wild rabbit populations in Australia. This study compared the infection and case fatality rates of RHDV2 and two RHDVs in wild rabbits, as well as their ability to overcome immunity to the respective other strains. Wild rabbits were allocated to groups either blindly or based on pre‐screening for RHDV/RHDV2 antibodies at capture. Rabbits were monitored regularly until their death or humane killing at 7 days post infection. Liver and eyeball samples were collected for lagovirus testing and aging rabbits, respectively. At capture, rabbits showed high seroprevalence to RHDV2 but not to RHDV. In RHDV/RHDV2 seronegative rabbits at capture, infection rates were highest in those inoculated with RHDV2 (81.8%, 18 out of 22), followed by K5 (53.8%, seven out of 13) and CZECH (40.0%, two out of five), but these differences were not statistically significant. In rabbits with previous exposure to RHDV2 at capture, infection rates were highest when inoculated with K5 (59.6%, 31 out of 52) followed by CZECH (46.0%, 23 out of 50), with infection rates higher in younger rabbits for both viruses. In RHDV/RHDV2 seronegative rabbits at capture, case fatality rates were highest for those inoculated with K5 (71.4%), followed by RHDV2 (50.0%) and CZECH (50.0%). In rabbits with previous exposure to RHDV2 at capture, case fatality rates were highest in rabbits inoculated with K5 (12.9%) followed by CZECH (8.7%), with no case fatalities following RHDV2 inoculation. Case fatality rates did not differ significantly between inoculums in either serostatus group at capture. Based on multivariable modelling, time to death post RHDV inoculation increased in rabbits with recent RHDV2 exposure compared with seronegative rabbits and with age. The results suggest that RHDV2 may cause higher mortalities than other variants in seronegative rabbit populations but that K5 may be more effective in reducing rabbit populations in an RHDV2‐dominant landscape. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease (RHD) is a highly contagious viral disease affecting lagomorphs. The first documented cases of RHD in Singapore occurred in adult pet European rabbits in September 2020. Singapore subsequently declared the outbreak resolved in December 2020. Epidemiological investigations ruled out introductions via importation of infected rabbits and contaminated feed. The source could not be definitively determined. However, the findings suggested that the incident involved both inter‐ and intra‐household transmission and veterinary clinic‐household transmission. This incident demonstrated the importance of sustained application of biosecurity measures, epidemiological investigations including active case finding, control measures such as expedient vaccine dissemination and risk communications. It showed that even without a wild lagomorph population, an urbanized city‐state like Singapore could still encounter emerging diseases such as RHD. Given its social impact on rabbit owners, the National Parks Board, Singapore and private veterinarians worked together to communicate with rabbit owners in order to urge them to adopt biosecurity measures and to address their concerns. [ABSTRACT FROM AUTHOR]

Simple Summary: Rabbit breeding is an important branch of agricultural animal breeding; their fur color and weight are desirable traits for artificial breeding. Polymorphism can provide potential molecular markers for studying rabbit traits and improve rabbit breeds with such markers in the future. In this study, single-nucleotide polymorphism markers in genotyping-by-sequencing data were used to analyze rabbit traits. In total, three genes were identified to be associated with fur color and four with weight. The results of this study provide a data base for the research and improvement of rabbit breeding program. The domestic rabbit (Oryctolagus cuniculus f. domesticus) is a very important variety in biomedical research and agricultural animal breeding. Due to the different geographical areas in which rabbit breeds originated, and the long history of domestication/artificial breeding, rabbits have experienced strong selection pressure, which has shaped many traits of most rabbit varieties, such as color and weight. An efficient genome-wide single-nucleotide polymorphism (SNP) detection strategy is genotyping-by-sequencing (GBS), which has been widely used in many organisms. This study attempted to explore bi-allelic SNPs associated with fur color and weight-related traits using GBS in five rabbit breeds. The data consisted of a total 831,035 SNPs in 150 individuals from Californian rabbits (CF), German Zika rabbits (ZK), Qixing rabbits (QX), Sichuan grey rabbits (SG), and Sichuan white rabbits (SW). In addition, these five breeds of rabbits were obviously independent populations, with high genetic differentiation among breeds and low genetic diversity within breeds. A total of 32,144 SNP sites were identified by selective sweep among the different varieties. The genes that carried SNP loci in these selected regions were related to important traits (fur color and weight) and signal pathways, such as the MAPK/ERK signaling pathway and the Hippo signaling pathway. In addition, genes related to fur color and weight were identified, such as ASIPs, MITFs and KITs, ADCY3s, YAPs, FASs, and ACSL5s, and they had more SNP sites. The research offers the foundation for further exploration of molecular genetic markers of SNPs that are related to traits. [ABSTRACT FROM AUTHOR]

Rabbit Haemorrhagic Disease Virus (RHDV), which is a calicivirus, is used as a biocontrol agent to suppress European wild rabbit populations in Australia. The transmission of RHDV can be influenced by social interactions of rabbits; however, there is a paucity of this knowledge about juvenile rabbits and the roles they may play in the transmission of RHDV. We aimed to quantify the social interactions of juvenile (< 900 g) and adult (> 1200 g) rabbits in a locally abundant population in the Central Tablelands of New South Wales, Australia. Twenty-six juvenile and 16 adult rabbits were fitted with VHF proximity loggers to monitor intra- and inter-group pairings. Use of multiple warrens by these rabbits was investigated using VHF base stations at nine warrens and on foot with a hand-held Yagi antenna. Juvenile rabbits were strongly interconnected with both juveniles and adults within and outside their warren of capture, and almost all juveniles were well-connected to other individuals within their own social group. Inter-group pairings were infrequent and fleeting between adults. Both juvenile and adult rabbits used multiple warrens. However, visits to warrens outside their warren of capture, particularly those within 50 m, were more common and longer in duration in juveniles than in adults. The high connectivity of juveniles within and between warrens in close proximity increases potential pathogen exchange between warrens. Therefore, juvenile rabbits could be of greater importance in lagovirus transmission than adult rabbits. The strength of juvenile rabbit inter- and intra-group pairings, and their tendency to use multiple warrens, highlight their potential to act as 'superspreaders' of both infection and immunity for lagoviruses and other pathogens with similar lifecycles. Confirmation of this potential is required through examination of disease progress and rabbit age-related immune responses during outbreaks. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus (RHDV) is associated with high morbidity and mortality in the European rabbit (Oryctolagus cuniculus). In 2010, a genetically distinct RHDV named RHDV2 emerged in Europe and spread to many other regions, including North America in 2016. Prior to this study it was unknown if eastern cottontails (ECT(s); Sylvilagus floridanus), one of the most common wild lagomorphs in the United States, were susceptible to RHDV2. In this study, 10 wild‐caught ECTs and 10 New Zealand white rabbits (NZWR(s); O. cuniculus) were each inoculated orally with either RHDV (RHDVa/GI.1a; n = 5 per species) or RHDV2 (a recombinant GI.1bP‐GI.2; n = 5 per species) and monitored for the development of disease. Three of the five ECTs that were infected with RHDV2 developed disease consistent with RHD and died at 4 and 6 days post‐inoculation (DPI). The RHDV major capsid protein/antigen (VP60) was detected in the livers of three ECTs infected with RHDV2, but none was detected in the ECTs infected with RHDV. Additionally, RHD viral RNA was detected in the liver, spleen, intestine and blood of ECTs infected with RHDV2, but not in the ECTs infected with RHDV. RHD viral RNA was detected in urine, oral swabs and rectal swabs in at least two of five ECTs infected with RHDV2. One ECT inoculated with RHDV2 seroconverted and developed a high antibody titre by the end of the experimental period (21 DPI). ECTs inoculated with the classic RHDV did not seroconvert. In comparison, NZWRs inoculated with RHDV2 exhibited high mortality (five of five) at 2 DPI and four of five NZWRs inoculated with RHDV either died or were euthanized at 2 DPI indicating both of these viruses were highly pathogenic to this species. This experiment indicates that ECTs are susceptible to RHDV2 and can shed viral RNA, thereby suggesting this species could be involved in the epidemiology of this virus. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease virus type 2 (RHDV2) causes a severe systemic disease with hepatic necrosis. Differently from classic RHDV, which affects only European rabbits (Oryctolagus cuniculus), RHDV2 can affect many leporid species, including hares (Lepus spp.) and cottontail rabbits (Sylvilagus spp.). RHDV2 emerged in Europe in 2010 and spread worldwide. During the last 5 years, there have been multiple outbreaks in North America since the first known event in 2016 in Quebec, Canada, including several detections in British Columbia, Canada, between 2018 and 2019, Washington State and Ohio, USA, in 2018 and 2019, and New York, USA, in 2020. However, the most widespread outbreak commenced in March 2020 in the southwestern USA and Mexico. In California, RHDV2 spread widely across several southern counties between 2020 and 2021, and the aim of this study was to report and characterize these early events of viral incursion and circulation within the state. Domestic and wild lagomorphs (n = 81) collected between August 2020 and February 2021 in California with a suspicion of RHDV2 infection were tested by reverse transcription quantitative real‐time PCR on the liver, and histology and immunohistochemistry for pan‐lagovirus were performed on liver sections. In addition, whole genome sequencing from 12 cases was performed. During this period, 33/81 lagomorphs including 24/59 domestic rabbits (O. cuniculus), 3/16 desert cottontail rabbits (Sylvilagus audubonii), and 6/6 black‐tailed jackrabbits (Lepus californicus) tested positive. All RHDV2‐positive animals had hepatic necrosis typical of pathogenic lagovirus infection, and the antigen was detected in sections from individuals of the three species. The 12 California sequences were closely related (98.9%–99.95%) to each other, and also very similar (99.0%–99.4%) to sequences obtained in other southwestern states during the 2020–2021 outbreak; however, they were less similar to strains obtained in New York in 2020 (96.7%–96.9%) and Quebec in 2016 (92.4%–92.6%), suggesting that those events could be related to different viral incursions. The California sequences were more similar (98.6%–98.7%) to a strain collected in British Columbia in 2018, which suggests that that event could have been related to the 2020 outbreak in the southwestern USA. [ABSTRACT FROM AUTHOR]

Rabbit haemorrhagic disease (RHD) is a significant viral disease caused by infection with Rabbit haemorrhagic disease virus (RHDV). The first documented cases of RHDV in Singapore occurred in adult pet European rabbits (Oryctolagus cuniculus) in September 2020. Rabbits presented with acute hyporexia, lethargy, huddled posture, and varying degrees of pyrexia and tachypnoea. Clinical pathology consistently reflected markedly elevated alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALKP). Hepatic lobe torsion was ruled out using ultrasonography and colour Doppler studies in all patients. A total of 11 rabbits owned by 3 families were presented to the clinics; 8/11 rabbits died within 48 hr of presentation, while the remaining two rabbits had recovered after prolonged hospitalization and one rabbit was aclinical. Histopathology revealed acute, marked diffuse hepatocellular necrosis and degeneration, findings which were suggestive for RHDV infection and prompted the undertaking of further molecular diagnostics. Subsequent polymerase chain reaction of the liver samples detected RHDV RNA. Molecular characterization of viral genomes by whole genome sequencing revealed that the outbreak strain was of the genotype GI.2 (RHDV2/RHDVb). Nucleotide sequences of the VP60 gene were compared with various RHDV variants using phylogenetic analysis. The sample genome shared highest sequence identity with a GI.2-genotyped virus from GenBank (RHDV isolate Algarve 1 polyprotein and minor structural protein (VP10) genes, GenBank accession KF442961). The combination of clinical, histopathological, molecular and sequencing technologies enabled rapid detection and detailed genetic characterization of the RHDV virus causing the present outbreak for prompt implementation of disease control measures in Singapore. Further epidemiological investigations of potential virus introduction into Singapore are ongoing. [ABSTRACT FROM AUTHOR]