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Background Rabbit haemorrhagic disease virus (RHDV) is a Lagovirus, a subgroup of the family Caliciviridae. RHDV2 is a variant first described in France in 2010, and has since spread globally. It has been reported in several Lagomorph species (rabbits, hares, and their relatives) as well as other mammals including voles and shrews. The disease has raised international concerns for its potential impact on population abundance trajectories, particularly as 25% of Lagomorphs are currently Red-Listed by the International Union for the Conservation of Nature (IUCN). The Irish hare (Lepus timidus hibernicus) is a subspecies of the mountain hare, L. timidus, and is endemic to Ireland, making it an Evolutionarily Significant Unit of intrinsic value. Case presentation The first case of RHDV2 was detected in a wild Irish hare in July 2019. The individual exhibited atypical neurological behaviour (running in circles) prior to death. On necropsy, pink tinged foam was seen in the trachea and congestion was noted in the lungs, but there was no evidence of haemorrhages in any other organ. Both the liver and spleen were tested by reverse transcription real time qPCR confirming high levels of RHDV2 RNA. Histopathology confirmed multifocal necrotising hepatitis. Conclusion The Irish hare is susceptible to RHDV2 infection. Further investigation is warranted to explore the clinical, epidemiological, and population biology implications.

Rabbit haemorrhagic disease virus (RHDV) was recovered from necropsied rabbits that died during an outbreak characterized by epistaxis, incoordination, paralysis, and multi-organ haemorrhages in Ilorin, Nigeria. The haemagglutination test (HA) and RT-PCR assay targeted against a fragment of the RHDV VP60 gene were performed on liver, spleen, and kidney homogenates; faeces; and urine obtained from the rabbits. Amplicons were purified, sequenced, and phylogenetically analysed. The liver homogenates yielded the highest HA titres while RT-PCR of liver, spleen, and kidneys yielded the expected 1252 bp band. Sequence and phylogenetic analyses revealed that the Nigerian RHDV strain (RHDV/NGR/ILN/001) was 98.57%, 97.95%, and 96.70% homologous with RHDV2 (RHDVGI.2) strains from the Netherlands, Germany, and France, respectively. RHDV/NGR/ILN/001 induced tracheal, intestinal, and mediastinal lymph node haemorrhages, pulmonary oedema and congestion, and enlarged, necrotic liver in experimentally inoculated rabbits. The implications of this study, which is the first report of RHDV in Nigeria, are discussed.

The introduced European rabbit (Oryctolagus cuniculus) is one of Australia’s most damaging invasive alien species, both in terms of ecological and economic impact. Biological control of rabbits using the myxoma and rabbit haemorrhagic disease viruses has been undertaken in Australia since the mid-1950s, and locally varying genetic resistance to these biocontrol viruses has been reported. The efficacy of biocontrol agents may be influenced, among several factors, by the genetic background of rabbit populations. Therefore, understanding the invasion process of rabbits in Australia, and their resultant population structure, remains crucial for enhancing future rabbit management strategies. Using reduced-representation sequencing techniques we genotyped 18 Australian rabbit populations at 7617 SNP loci and show that Australia’s invasive rabbits form three broad geographic clusters representing different ancestral lineages, along with a number of highly localised, strongly differentiated lineages. This molecular data supports a history of multiple independent rabbit introductions across the continent followed by regional dispersal, and the resulting patchwork genetic structure may contribute to variation across the country in rabbit resistance to the viral biocontrols. Our study highlights the importance of using genome-wide molecular information to better understand the historical establishment process of invasive species as this may ultimately influence genetic variabilty, disease resistance and the efficacy of biocontrol agents.

The rabbit haemorrhagic disease virus (RHDV) was imported into Australia in 1995 as a biocontrol agent to manage one of the most successful and devastating invasive species, the European rabbit (Oryctolagus cuniculus cuniculus). During the first disease outbreaks, RHDV caused mortality rates of up to 97% and reduced Australian rabbit numbers to very low levels. However, recently increased genetic resistance to RHDV and strong population growth has been reported. Major histocompatibility complex (MHC) class I immune genes are important for immune responses against viruses, and a high MHC variability is thought to be crucial in adaptive processes under pathogen-driven selection. We asked whether strong population bottlenecks and presumed genetic drift would have led to low MHC variability in wild Australian rabbits, and if the retained MHC variability was enough to explain the increased resistance against RHD. Despite the past bottlenecks we found a relatively high number of MHC class I sequences distributed over 2–4 loci. We identified positive selection on putative antigen-binding sites of the MHC. We detected evidence for RHDV-driven selection as one MHC supertype was negatively associated with RHD survival, fitting expectations of frequency-dependent selection. Gene duplication and pathogen-driven selection are possible (and likely) mechanisms that maintained the adaptive potential of MHC genes in Australian rabbits. Our findings not only contribute to a better understanding of the evolution of invasive species, they are also important in the light of planned future rabbit biocontrol in Australia., This study was funded by the Priority Programme of the German Science Foundation (DFG) ‘Host-parasite co-evolution—rapid reciprocal adaptation and its genetic basis’ (SPP 1399, PI: So 428/7-1).

The arrival of a new variant of rabbit haemorrhagic disease virus, known as RHDV2, has recently taken place in the native range of the European rabbit (Oryctolagus cuniculus), a keystone species which has undergone a sharp decline over the last sixty years as a consequence of certain harmful factors. Several works have noted the presence of this new variant in wild rabbit populations, and have in some cases recorded high mortality rates. However, little is known about the response to the arrival of this new virus variant at the population level. The goal of this work is therefore to show recent trends in 26 wild rabbit populations between 2010 (before the outbreak of the disease) and 2014 (after its onset) in two different ecosystems (woodland and agricultural areas), in order to test how their abundances changed over this period, which coincided with the spread of the RHDV2. Overall, our results showed that rabbit abundance was much lower in 2014 than in 2010, and that only 11.5% of the populations monitored proved to have a positive trend, that is, a higher abundance in 2014 than 2010. A positive correlation between rabbit abundance in 2010 and rabbit population trends was obtained, thus suggesting that the impact of the new variant on rabbit abundance is less evident in high density populations. Our results suggest that smaller rabbit populations are those most vulnerable to the outbreak of RHDV 2 and are therefore likely to decline sharply or even become extinct.

Background In the regular wildlife monitoring action carried out in the summer of the past few years at the Berlenga Island, wild rabbits (Oryctolagus cuniculus) have been repeatedly found dead. However, the origin of those deaths was never investigated. Our aim was to investigate the cause of death of 11 rabbits collected between April and May 2016. Results While screening samples from rabbit carcasses for the major viral rabbit pathogens, five tested positive to RHDV2 but all were negative for RHDV and myxoma virus (MYXV). For six RHDV2-negative specimens, emaciation and parasitism were considered the most probable cause of death. Lesions identified in the RHDV2-positive rabbits included non-suppurative diffuse hepatic necrosis and pulmonary lesions varying from congestion and oedema of the lungs to interstitial pneumonia. Sequencing analysis of the vp60 gene obtained from two specimens showed identical vp60 sequences. Comparison with other known RHDV2 strains from public databases through BLAST analysis revealed a closer similarity with strains from Alentejo collected during 2013. Maximum Likelihood and Bayesian phylogenetic analysis showed that the 2016 strains from the archipelago have a higher resemblance with a group of strains mostly collected in the South of Portugal between 2013 and 2014. Conclusion The results suggest that RHDV2 may have been introduced on the Berlenga Island a few years ago, having evolved separately from mainland strains due to insularity.

Rabbit haemorrhagic disease virus (RHDV) is a lagovirus that causes rabbit haemorrhagic disease (RHD) in European rabbits (Oryctolagus cuniculus). In 2010, a new genotype called RHDV2 emerged in France. It exhibits a larger host range than classical RHDV strains by sporadically infecting different hare species, including the European hare (Lepus europaeus). Phylogenetic analyses revealed that closely related RHDV2 strains circulate locally in both hares and rabbits, and therefore that RHDV2 strains infecting hares do not belong to a lineage that has evolved only in this species. We showed that RHDV2 is widely distributed in France and that it was responsible for more than a third of cases of lagovirus disease in European hare populations in 2015. The oldest RHDV2 positive hare was sampled in November 2013 and we reported two hares co-infected by EBHSV and RHDV2. All together, our results raise important epidemiological and evolutionary issues. In particular, along with the potential emergence of recombinant EBHSV/RHDV2 strains in hares, the enlargement of the host range changes the host population structure of RHDV2 and may alter the impact of the virus on rabbit and hare populations. Electronic supplementary material The online version of this article (doi:10.1186/s13567-017-0473-y) contains supplementary material, which is available to authorized users.

The frequency and timing of rabbit haemorrhagic disease (RHD) epizootics and their impact on different age groups of rabbits were studied for 15 years in a recovering rabbit population in South Australia. We recorded the number and body size of rabbits dying during RHD epizootics, collected tissue for genetic analysis of rabbit haemorrhagic disease virus variants and compared the number of carcasses found to the number of susceptible rabbits present at the beginning of each epizootic. All RHD epizootics occurred between late winter and spring, but, progressively, epizootics started earlier and became more frequent and prolonged, fewer susceptible adult rabbits were present during epizootics, and the age of rabbits dying of RHD declined. Increased infection and virus shedding in juvenile rabbits offers the most plausible explanation for those epidemiological changes; the disease is now increasingly transmitted through populations of kittens, starting before young-of-the-year reach adult size and persisting late in the breeding season, so that most rabbits are challenged in their year of birth. These changes have increased juvenile mortality due to RHD but reduced total mortality across all age groups, because age-specific mortality rates are lower in young rabbits than in older rabbits. We hypothesise that this may be the proximate cause of recovery in rabbit populations across Australia and possibly elsewhere.

Rabbit haemorrhagic disease virus (RHDV), genus Lagovirus, family Caliciviridae, causes a large number of deaths in wild and domestic adult European rabbits (Oryctolagus cuniculus). The first documented outbreak dates from 1984 in China, but the virus rapidly dispersed worldwide. In 1997, an antigenic variant was detected in Italy and designated RHDVa. Despite causing symptoms similar to those caused by classic RHDV strains, marked antigenic and genetic differences exist. In some parts of Europe, RHDVa is replacing classic strains. Here, we report the presence of RHDVa on the Iberian Peninsula, where this variant was thought not to contribute to viral diversity.

European brown hare syndrome (EBHS) is characterised by high mortality of European brown hares (Lepus europaeus) and mountain hares (Lepus timidus). European brown hare syndrome virus (EBHSV) and the closely related rabbit haemorrhagic disease virus (RHDV) comprise the genus Lagovirus, family Caliciviridae. In contrast to RHDV, which is well studied, with more than 30 complete genome sequences available, the only complete genome sequence available for EBHSV was obtained from a strain isolated in 1989 in France. EBHS was originally diagnosed in Sweden in 1980. Here, we report the complete coding sequences of two EBHSV strains isolated from European brown hares that died with liver lesions characteristic of EBHS in Sweden in 1982. These sequences represent the oldest complete coding sequences of EBHSV isolated from the original area of virus diagnosis. The genomic organisation is similar to that of the published French sequence. Comparison with this sequence revealed several nucleotide substitutions, corresponding to 6 % divergence. At the amino acid level, the Swedish strains are 2 % different from the French strain. Most amino acid substitutions were located within the major capsid protein VP60, but when considering the amino acid sequence length of each protein, VP10 is the protein with the highest percentage of amino acid differences. The same result was obtained when Swedish strains were compared. This evolutionary pattern has not been described previously for members of the genus Lagovirus.

In a previous study, we suggested that hyperpredation by shared predators on red-legged partridges (Alectoris rufa), once a parasite (rabbit haemorrhagic disease, RHD) had decimated populations of rabbit (Oryctolagus cuniculus), the primary prey for most of the Mediterranean predators, was a major force driving partridge population dynamics in Spain years ago (Moleon et al., PLoS One 3:e2307, 2008). Recently, however, Blanco-Aguiar et al. (Eur J Wildl Res 58:433–439, 2012) have asserted that this conclusion is poorly evidence-based and can subsequently promote raptor persecution by hunters. In response to Blanco-Aguiar and colleagues, here we provide complementary insights that favour our earlier hypothesis. After explaining several key concepts of the hyperpredation process, we use additional data and analyses to show that (1) a synchronised regime shift (i.e. a step, abrupt change in population size) to significantly lower population levels in both the rabbit and partridge populations took place coinciding with the RHD outbreak; (2) rabbit and partridge population dynamics were highly synchronised after the RHD outbreak, but not before; (3) an enhanced spatial autocorrelation at all the spatial scales emerged after RHD for partridge populations; and (4) the main shared predators’ diet patterns were consistent with the hypothesis of enhanced predation pressure as a plausible mechanism behind the observed partridge dynamics. We support the idea that hunting bag data may be useful to infer realistic population dynamics and the ecological mechanisms explaining them, provided that (1) they are corrected by the number of hunting licenses; (2) appropriate statistical tools are employed; and (3) methodological constraints are adequately taken into account. Finally, we argue that the opinion of Blanco-Aguiar and colleagues that our original results can lead to raptor persecution is a misinterpretation of our study. In this sense, we make an appeal for the importance of accurately differentiating between the ultimate (e.g. infectious diseases favoured by humans) and the proximate (e.g. enhanced predation pressure) causes of the decline in prey of economic interest (e.g. game species) in order to avoid unnecessary, unfounded or presumed conflicts with lobbies of conservation concern (e.g. hunters).

The impact of over-abundant exotic herbivores is well recognised, but their impact at low population densities is poorly understood. This study examined interactions between European rabbits and native herbivores, and their impact on seedling recruitment in coastal South Australia, 2 years after rabbit haemorrhagic disease (RHD) had reduced rabbit density to 4.48 rabbits ha−1. Rabbit density was further reduced to 0.44 rabbits ha−1 in replicated experimental treatments. Rabbit control reduced total grazing pressure by 39% despite compensatory grazing increases of >100% for both western grey kangaroos and common wombats. Rabbit control slowed the rate of grazing and mortality for planted drooping sheoak and sweet bursaria seedlings, but few survived for 12 months: 0 and 3% of sheoak, in untreated areas and rabbit control treatments, respectively, and 3 and 11% of bursaria, respectively. Planted sheoaks survived well if protected by rabbit-proof netting (60%). Within treatments, seedling grazing and survival rates were negatively correlated with rabbit density but kangaroo and wombat density had no measurable effect. We conclude that RHD may briefly have reduced rabbit densities enough to allow recruitment of bursaria but that sheoak require much lower rabbit densities than those provided by existing biological control agents. If left unaddressed, rabbit grazing could ultimately lead to the loss of sheoaks throughout most of their current range, irrespective of other attempts to conserve them. More generally, these data show how species-specific damage caused by low-density exotic herbivore populations may occur in the presence of more abundant but less-damaging native herbivores.

Antibodies raised against a Lagovirus were found in healthy rabbits Oryctolagus cuniculus sampled in 2003 and 2004 in the Kerguelen archipelago. The serological test we used enabled the detection of antibodies due to both pathogenic and non-pathogenic viruses related to the rabbit haemorrhagic disease virus (RHDV). The overall proportion of seropositive rabbits was 35% and differed between sites. Since previous studies have failed to detect antibodies raised against pathogenic RHDV strains, the antibodies detected in the present study were likely due to non-pathogenic strains. The interest of these strains circulating in the Kerguelen archipelago is that they have evolved independently of those detected in the other parts of the world and should derive from an ancestral RHDV precursor. Their characterization may help understanding the evolution of the virus and the emergence of the disease.

A sparse rabbit population in New Zealand was monitored over 3 years to assess the temporal dynamics of rabbit mortality, in particular to understand the mortality patterns due to rabbit haemorrhagic disease (RHD). A total of 107 deaths were recorded, of which 93 could be classified by cause. The predominant cause of mortality was predation (47% of deaths), followed by RHD (20%). Deaths due to RHD were clustered in time (within 3 weeks), predation occurred most actively from late autumn to spring, while other causes of death did not show pronounced seasonal peaks. No differences in cause-specific death risk were observed between sexes. Predation was the main cause of death in younger animals, while RHD mortality occurred mainly in older rabbits. This study has shown that the impact of RHD can vary considerably between years, indicating that a variety of risk factors are required to initiate a RHD epidemic with a high mortality rate among rabbits.

Rabbit haemorrhagic disease (RHD) is a highly fatal disease caused by a virus of the family Caliciviridae. Whereas recombination is well documented in other members of this family, the extent of recombination has so far not been studied in RHDV. To reach a better evaluation of the possible role of recombination in the evolution of RHDV virulence, we have searched for recombination events in RHDV by analysing 43 complete sequences of the major capsid gene VP60. Phylogenetic analyses revealed two well separated groups. Clear evidence for recombination was found for the Hartmannsdorf strain which shows different phylogenetic profiles depending on the region of the capsid examined.

Rabbit haemorrhagic disease (RHD) is caused by a calicivirus infection that kills most adult rabbits 24-72 h after viral inoculation. Two liver enzymes (AST, aspartate aminotransferase, and ALT, alanine aminotransferase) were monitored in blood samples of calicivirus-infected rabbits during the short course of RHD. Values of AST were used to differentiate three stages of hepatocellular degeneration in RHD: mild (up to 20-fold increase in AST), moderate (150-200-fold elevation of AST) and severe (more than 1000-fold elevation in AST). Liver samples of rabbits from these three biochemical stages of hepatocellular degeneration of RHD were studied by transmission electron microscopy to define the fine structure of the hepatocytes. In the mild hepatocellular degeneration there was proliferation (microvesiculation) of the smooth endoplasmic reticulum and swelling of mitochondria into spheroid bodies with loss of cristae. In moderate hepatocellular degeneration, vacuolization of cytoplasm and mitochondrial damage continued to be present, and there was also formation of autophagic vesicles. In the severe hepatocellular degeneration of RHD, the altered mitochondria also showed loss of density of their matrix; rupture of cytoplasmic vacuoles led to the formation of large vesicles. Marked depletion of liver glycogen was also found in this late stage of RHD. These data offer a correlation between biochemical and cytological features of the liver during the hepatocellular degeneration of RHD.

International audience; Genetic diversity between French European brown hare syndrome (EBHS) viruses since the disease appeared has been evaluated. Nucleotide sequencing of the partial capsid protein genes of 169 EBHS viruses collected from various parts of France between 1989 and 2003, three reference strains, and a Greek EBHSV collected in 2002 revealed a maximum nucleotide divergence of 11.7%, indicating a high level of conservation between viruses. Two major groups were identified. The first group contained EBHS viruses collected since 1989 from different parts of France, the reference strains, and all of the viruses located in the far north of France. In this group, three genogroups were clearly identified as mainly related to their geographic origin. The distribution of the viruses suggests that the early viruses have not disappeared and have slowly evolved in their area of origin. The second group, supported by a significant bootstrap value, contained the Greek EBHSV with the French EBHS viruses collected between 1999 and 2003 from regions of southern France. It constitutes a newly identified genogroup. Our results demonstrate strong differences in genetic evolution between EBHSV and rabbit haemorrhagic disease virus, with persistence of the earlier EBHS viruses and interaction between the geographical and temporal distributions.

To define the genetic variability of RHDV strains collected in eastern Hungary, liver samples from rabbits that had died of RHD were collected between 1988 and 2003. The phylogenetic analysis of a 528-nucleotide-long portion of the gene encoding the VP60 capsid protein assigned the strains into three genogroups. The first group contained viruses from 1988-1993, and a second group comprised isolates from 1994-2002. A third group comprised all of the tested representatives of the RHDVa subtype and a Hungarian isolate from 2003. These findings were supported by the alignments of the deduced amino acid sequences of the VP60 gene and strongly suggest the presence of the RHDVa subtype in Hungary.

Rabbit haemorrhagic disease virus (RHDV) and Pasteurella multocida bacteria cause severe losses among rabbit populations. The efficacy of a recently developed bivalent vaccine against pasteurellosis and RHDV was investigated. Doses exceeding 2 haemagglutinating units (HU) of viral antigen were sufficient to protect rabbits against infection with RHDV. The bivalent vaccine appeared to be safe for use in all age groups of rabbits, including pregnant females, even after treatment with 20 times the normal vaccine dose. Rabbits injected with 8 or 4 HU of bivalent vaccine showed high antibody titres against both organisms for 9 months after inoculation. The antibody levels against RHDV in young rabbits at 30 days of age were elevated when they originated from mothers with high antibody titres. The most suitable period for vaccination of offspring appeared to be around 50 days of age. The bivalent vaccine against pasteurellosis and RHDV combined speed and longevity of the immune response. Immune protection against pasteurellosis and RHDV can thus be achieved with only one manipulation.

The Eagle Owl (Bubo bubo) is a generalist predator that in Mediterranean areas feeds mainly on Rabbits (Oryctolagus cuniculus) At the end of 1997, a local outbreak of the rabbit haemorrhagic disease (RHD) decimated Rabbit populations in the area of Alicante (eastern Spain) so that Rabbit numbers in 1998 crashed to almost nil. Prior to the outbreak we had found 19 occupied Eagle Owl territories, and the owls had been feeding mainly on Rabbits. After the RHD epidemic, we found only six occupied territories and the owls were still feeding mainly on Rabbits. Diet composition was very similar between the periods, indicating that the owls did not diversify their diet in response to main prey scarcity. No significant differences in the proportion of Rabbit in the diet were found between the periods, suggesting that the impact of predation on Rabbits may be independent of the density of Rabbit populations (type I functional response). This response is not typically associated with generalist predators. In the territories that were still occupied after the outbreaks, the owls took substantial numbers of Hedgehogs (Erinaceus europaeus) and Red-legged Partridges (Alectoris rufa). Rats (Rattus spp), a common alternative prey, were not taken in the study area. Six breeding attempts were recorded before the RHD, and none after the RHD. This leads a to believe that only the combination of Hedgehog and Red-legged Partridge availability with the readier availability of sick Rabbits allowed a small population of owls to survive in the area, but did not allowed breeding. The predator-prey system formed by Eagle Owls and Rabbits in Mediterranean regions is more fragile than previously thought.

Sera collected from Australian wild rabbits prior to the escape of rabbit haemorrhagic disease virus (RHDV) from Wardang Island were examined for RHDV antibodies using purified recombinant capsid protein VP60 expressed from baculovirus. A VP60-based indirect ELISA showed that 196 of 392 wild rabbit sera reacted (OD(450) >0.15) with VP60. Twenty sera (OD(450) ranging from 0.15-2.47), randomly chosen from the 196 positive sera, recognized the 64 kDa VP60 in Western blot analysis, indicating that the reactivity detected in ELISA is indeed specific to the VP60 antigen. In a separate study, sera of 23 rabbits from an RHD-free area after the escape of RHDV were tested by ELISA and 21 of the 23 rabbits were found to be positive. When these rabbits were challenged with a lethal dose of RHDV, 11 out of the 23 rabbits survived. The presence of RHDV-protective antibodies in some of these rabbits suggested that they had been exposed to a pre-existing non-virulent rabbit calicivirus closely related to RHDV. These results highlight the need to study the prevalence of, and to characterize, this viral agent in order to effectively control rabbit populations in Australia and New Zealand.

A reverse transcription-polymerase chain reaction (RT-PCR) amplification assay was used to detect calicivirus gene sequences in a liver tissue derived from a feral rabbit which died of a recent outbreak of rabbit haemorrhagic disease (RHD) in New Zealand. Five pairs of primers were designed to amplify five complementary DNA genomic sequence stretching from nucleotide positions 1594 to 7071, yielding amplified fragments of 361, 340, 805, 670 and 386 bp for the primer pairs RC-1/RC-2, RC-3/RC-4, RC-5/RC-6, RC-7/RC-8 and RC-9/RC-10 respectively. The identity of the amplified fragments was confirmed by chemiluminescence Southern blot hybridization and direct cycle sequencing. The nucleotide sequences of the five amplified fragments were determined and comparisons of the nucleotide and deduced amino acid sequences revealed a close genetic relationship of the New Zealand isolate 97-10372 with overseas strains of RHD virus.

A 398 bp fragment of the capsid protein (VP60) gene of 39 clinical samples of rabbit haemorrhagic disease virus (RHDV) and 17 of European brown hare syndrome virus (EBHSV), collected between 1981 and 1995 from 17 countries, was amplified by PCR and directly sequenced. The alignment of the nucleotide sequences and the subsequently constructed phylogenetic tree clearly separated RHDV from EBHSV as phylogenetic entities. The nucleotide homology rates between the RHDV and EBHSV groups ranged between 52.6% and 60.0%. The homology rates within the groups were much higher, 89.4% to 100% for the RHDV samples, and 89.4% to 100% for the EBHSV specimens. No intermediate viruses were found. Despite the high homology, three main branches could be identified in the phylogenetic tree of the RHDV samples, corresponding to the epizootiological data, while the EBHSV dendrogram did not show such well defined branches. The present results support the classification of RHDV and EBHSV as two distinct members of the Caliciviridae family. Nevertheless, a comparison with previously determined sequences of other caliciviruses shows that RHDV and EBHSV are more closely related to each other than to any other calicivirus.

The virus recovered from cases of European brown hare syndrome in the U.K. contains one major capsid protein of approximately 60 k molecular weight and morphologically resembles known caliciviruses. It has been compared with a European isolate of rabbit haemorrhagic disease calicivirus and, although it shows some antigenic similarity, it is not identical. In transmission and protection studies the virus from U.K. hares failed to produce disease in rabbits and did not effectively protect against subsequent challenge with the rabbit calicivirus.

Immunohistochemical investigations were carried out to determine organ and cellular localization of the rabbit haemorrhagic disease viral antigen (RHDVA). It was found in certain parenchymal liver cells near the interlobular septs and in some macrophages and pseudoeosinophils of all studied organs and blood. Whereas in morphologically preserved hepatocytes and macrophages the RHDVA accumulated in the nuclei, in cells with disintegrated nuclei it was distributed throughout the cytoplasm.

Rabbit haemorrhagic disease virus (RHDV) had a calicivirus-like structure and a diameter of 31.5-33.0 nm. Antigenic relationship between the investigated RHDV strain and the causal agent of RHD in China was demonstrated by immunoelectron microscopy.

International audience; Since summer 2010, numerous cases of Rabbit Haemorrhagic Disease (RHD) have been reported in north-western France both in rabbitries, affecting RHD-vaccinated rabbits, and in wild populations. We demonstrate that the aetiological agent was a lagovirus phylogenetically distinct from other lagoviruses and which presents a unique antigenic profile. Experimental results show that the disease differs from RHD in terms of disease duration, mortality rates, higher occurrence of subacute/chronic forms and that partial cross-protection occurs between RHDV and the new RHDV variant, designated RHDV2. These data support the hypothesis that RHDV2 is a new member of the Lagovirus genus. A molecular epidemiology study detected RHDV2 in France a few months before the first recorded cases and revealed that one year after its discovery it had spread throughout the country and had almost replaced RHDV strains. RHDV2 was detected in continental Italy in June 2011, then four months later in Sardinia.