Belgian Wildlife as Potential Zoonotic Reservoir of Hepatitis E Virus

Hepatitis E is an acute human liver disease in healthy individuals but may become chronic in immunocompromised patients. It is caused by the hepatitis E virus (HEV) and can have a zoonotic origin, particularly in high‐income countries. In this study, 383 sera from wild boars were selected for serology; for virological analyses, 69 sera and 61 livers from young wild boars were used. A total of 189 and 235 sera of, respectively, red deer and roe deer were collected for serological analysis. For virological analyses, 84 and 68 sera and 29 and 27 livers from, respectively, red and roe deer were sampled. An apparent seroprevalence of 34% (95% CI 29.71–39.46) was found in wild boars, of 1% (95% CI 0–2.4) in red deer and 3% (95% CI 0.8–4.2) in roe deer. To assess the ELISA screening prevalence, Western blot (WB) analyses were carried out, a receiver operating characteristic curve analysis was performed and different scenarios with varying ELISA specificities relative to WB were analysed. Seroprevalence remained high whatever the scenario in the wild boar population. In wild boar, 4 of 69 sera and 4 of 61 livers were detected as positive for HEV RNA. All sequences obtained from sera belonged to genotype HEV‐3. HEV RNA, belonging to genotype HEV‐3, was detected in one of 29 red deer livers. Wild boar can be considered as a host reservoir of the virus in Belgium. However, in contrast to the epidemiological role played by them in other countries, the low prevalence in deer makes these species an unlikely reservoir. This evidence needs further investigation to determine in which situation deer can serve as reservoir. These results also raise the question of the dynamics of HEV infection between wild fauna, domestic pigs and humans.     

Constant Hepatitis E Virus (HEV) Circulation in Wild Boar and Red Deer in Spain: An Increasing Concern Source of HEV Zoonotic Transmission

Hepatitis E is a viral zoonosis that affects multiple hosts. The complete dynamics of infection in wildlife are still unknown, but the previous fact facilitates the maintenance and circulation of the virus, posing a risk to human health in the case of meat consumption from susceptible animals. In Spain, it has been shown how domestic pigs, cattle and wildlife (i.e. wild boar and red deer) clearly interact in hunting farms, generating a complex epidemiological situation in terms of interspecies pathogen transmission. Therefore, in this study, we aimed to (i) evaluate the circulation of the virus in geographically close domestic (Iberian pigs) and wild animals (wild boar and deer) living in hunting areas from central Spain over an 8‐year period (2003–2010) and (ii) to determine whether HEV could be used as a marker of domestic–wildlife contact. For these purposes, a longitudinal analysis of Iberian pig, wild boar and red deer samples (n = 287) through virological and serological tests was conducted to shed light upon the circulation events of HEV. Regarding HEV RNA detection by real‐time RT‐PCR, 10.12% samples (95% CI: 5.44–14.8) from wild boar and 16.05% samples (95% CI: 8.06–24.04) from red deer were positive. As for the Iberian pigs, none of the 48 samples was positive for HEV RNA detection. In the serological analysis, 43.75% (95% CI: 29.75–57.75) from Iberian pig, 57.40% (95% CI: 48.10–66.70) from wild boar and 12.85% (95% CI: 5.01–20.69) samples from red deer presented anti‐HEV antibodies. Positive samples were distributed among all study years (2003–2010). These results depict the urgent need to improve the inspection and surveillance of these species and their products. In the case of HEV, it is clear that the stable and constant presence of the virus in wildlife and its contact with Iberian pigs pose a risk for human health as they are all destined for human consumption.     

Hepatitis E Virus Seroprevalence in Free‐Ranging Deer in Canada

Hepatitis E virus infection (HEV) is an important public health concern not only in traditional endemic areas, but also in some industrialized countries where both domesticated and wild animals have been recognized as potential zoonotic reservoirs implicated in HEV transmission. While the prevalence of infection in the deer population in Europe and Asia has been thoroughly investigated, it remains largely undetermined in North America. We assessed the presence of HEV in three different species of free‐range deer in Canada. The seroprevalence of HEV among deer in Canada was 8.8% in white‐tailed deer, 4.5% in mule deer and 3.2% in caribou. Hepatitis E virus RNA was not detected. Overall, data indicate that HEV infection occurs in deer in Canada. The absence of viraemia and the low seroprevalence especially in barren‐ground caribou which is an important part of the diet in many northern communities suggests that the risk of zoonotic transmission may be less pronounced compared to other countries.     

Serological Evidence for a Hepatitis E Virus‐Related Agent in Goats in the United States

Hepatitis E virus (HEV) causes an important public health disease in many developing countries and is also endemic in some industrialized countries. In addition to humans, strains of HEV have been genetically identified from pig, chicken, rat, mongoose, deer, rabbit and fish. While the genotypes 1 and 2 HEV are restricted to humans, the genotypes 3 and 4 HEV are zoonotic and infect humans and other animal species. As a part of our ongoing efforts to search for potential animal reservoirs for HEV, we tested goats from Virginia for evidence of HEV infection and showed that 16% (13/80) of goat sera from Virginia herds were positive for IgG anti‐HEV. Importantly, we demonstrated that neutralizing antibodies to HEV were present in selected IgG anti‐HEV positive goat sera. Subsequently, in an attempt to genetically identify the HEV‐related agent from goats, we conducted a prospective study in a closed goat herd with known anti‐HEV seropositivity and monitored a total of 11 kids from the time of birth until 14 weeks of age for evidence of HEV infection. Seroconversion to IgG anti‐HEV was detected in seven of the 11 kids, although repeated attempts to detect HEV RNA by a broad‐spectrum nested RT‐PCR from the faecal and serum samples of the goats that had seroconverted were unsuccessful. In addition, we also attempted to experimentally infect laboratory goats with three well‐characterized mammalian strains of HEV but with no success. The results indicate that a HEV‐related agent is circulating and maintained in the goat population in Virginia and that the goat HEV is likely genetically very divergent from the known HEV strains.     

Hepatitis E virus in sheep in Italy

Hepatitis E virus (HEV) is the leading cause of human enterically transmitted viral hepatitis occurring around the world both as outbreaks and as sporadic cases. The accumulating literature indicates that domestic pigs and wild boars are the main reservoirs of genotype 3 and genotype 4 for human infections in industrialized countries. However, the recent identification of HEV from various animal species poses additional potential concerns for HEV zoonotic infection. In this study, the role of sheep as potential host of hepatitis E virus (HEV) was investigated. By screening 192 sheep from seven farms located in Abruzzo Region (Southern Italy), HEV‐specific antibodies were detected in the sera of 41 animals (21.3%) whilst the RNA of HEV, genotype 3, was detected in 20 faecal (10.4%) and three serum samples (1.6%). Upon sequence analyses of a partial ORF2 gene region of eight HEV positive samples, the sheep sequences all grouped together within HEV genotype 3 subtype c, being most closely related to HEV strains identified in goat and wild boar from Abruzzo. This is the first study that demonstrates, serologically and molecularly, the presence of HEV in sheep population in a European country.     

Molecular detection and phylogenetic analysis of hepatitis E virus strains circulating in wild boars in south‐central Italy

Hepatitis E virus (HEV) is a zoonotic pathogen with a worldwide distribution, and infects several mammalian species, including pigs and wild boars, which are recognized as its natural reservoirs. The virus causes a usually self‐limiting liver disease with a mortality rate generally below 1%, although mortality rates of 15%–25% have been recorded in pregnant woman. Chronic infections can also occur. The prevalence of HEV has been extensively studied in wild boars and pigs in northern Italy, where intensive pig herds are predominantly located. In contrast, few data have been collected in south‐central Italy, where small pig herds are surrounded by large regional parks populated with heterogeneous wild fauna. In this study, 291 liver samples from wild boars caught in south‐central Italy were analysed with the molecular detection of viral RNA. Our results confirm the circulation of HEV in these animals, with a mean prevalence of 13.7% (40 of 291). A nucleotide sequence analysis showed that the HEV strains were highly conserved within the same geographic areas. The wild boar HEV strains belonged to the HEV‐3c subtype, which is frequently described in wild boars, and to an uncommon undefined subtype (HEV‐3j‐like).The viral prevalence detected is concerning because it could represent a potential risk to hunters, meat workers and consumers of wild boar liver and derivative products. The hypothesized inter‐species transmission of HEV to pigs and the possibility that the virus maintains its virulence in the environment and the meat chain also present potential risks to human health, and warrant further investigations in the near future.     

Hepatitis E Virus: A Cross‐Sectional Serological and Virological Study in Pigs and Humans at Zoonotic Risk within a High‐Density Pig Farming Area

An increase in autochthonous hepatitis E virus (HEV) infections has been recorded in Italy suspected to be zoonotically transmitted from pigs; this study was carried out to determinate the seroprevalence and risk factors associated with hepatitis HEV exposition, both in swine and humans working in pig farms, located within a high‐density pig farming area in Piedmont region, north‐western Italy. The presence of viral RNA in human and swine samples was also evaluated, and phylogenetic analysis was performed on HEV‐positive samples. Forty‐two swine farms were sampled; 142 workers were enrolled in the study and classified into two groups: (i) 69 workers with occupational contact with swine (including veterinarians and farmers) recruited in the 42 sampled farms; (ii) 73 without occupational contact with swine. Forty‐one of 42 (97%) swine farms resulted positive to enzyme‐linked immunosorbent assay test for HEV antibodies (Abs). Overall seroprevalence in swine was 50% (441/879), with seropositivity rate higher in sows (333/469, 71%). HEV RNA in stool samples was detected in animals from 13 of 42 tested farms (31%), and a higher positivity resulted in weaners (40/246, 16.3%). Phylogenetic analysis classified all HEV isolates within genotype 3 (subtypes 3f, 3e, 3c). All humans were negative for HEV viral genome in blood. Five of 142 sera were positive for IgG anti‐HEV with an overall prevalence of 3.52% with no statistically significant differences in prevalence rates between workers at zoonotic risk and the control group (5.7% versus 1.3%). In contrast, a significant difference (OR 10.1) was observed within the subgroup including subjects exposed for short periods (veterinarians) compared with those who worked for long periods (farmers) suggesting a correlation between the time of exposure and the likelihood of HEV infection. Reporting HEV infection is not mandatory in Italy, but a constant epidemiological surveillance should be ensured to clarify the epidemiology of this disease.     

Survey for Hepatitis E virus infection in non‐human primates in zoos in Spain

Hepatitis E virus (HEV) is an emerging zoonotic pathogen that has been detected in different animal species. A survey study was carried out to assess HEV infection in non‐human primates (NHPs) housed in zoos in Spain. Anti‐HEV antibodies were detected in eight of the 181 NHPs tested (4.4%; 95%CI: 1.4–7.4). At least one seropositive animal was detected in five of the 33 species sampled (15.2%). This is the first report of seropositivity in black‐and‐white ruffed lemurs (Varecia variegata), common chimpanzees (Pan troglodytes), and Barbary macaques (Macaca sylvanus). Anti‐HEV antibodies were found in six of the eight zoos included in the study (75.0%). Seroconversion was detected in one chimpanzee, which confirms HEV circulation in one zoo between 2015 and 2016. Seropositivity was significantly higher in hominids than in other NHP families. HEV RNA was not detected in any of the serum samples tested. The results indicate susceptibility of NHPs to HEV infection. Further studies are required to elucidate the role of these species in the epidemiology of HEV.     

Hepatitis E Virus‐Induced Neurological Symptoms in a Kidney‐Transplant Patient with Chronic Hepatitis

It has been shown that hepatitis E virus (HEV) may be responsible for chronic hepatitis in solid‐organ transplant patients. It has also been suggested that HEV may be responsible for atypical neurological symptoms during the acute phase. However, the relationship between the neurological symptoms and HEV infection was based on the detection of anti‐HEV IgM in the sera. Herein, we report a case where neurological symptoms, that is peripheral nerve involvement with proximal muscular weakness that affected the four limbs joints with central nervous‐system involvement and bilateral pyramidal syndrome, occurred in a kidney‐transplant patient who was chronically infected by HEV. For the first time, HEV RNA was detected in the serum and cerebrospinal fluid. In addition, clonal HEV sequences were analyzed in both compartments, that is serum and cerebrospinal fluid. The discovery of quasispecies compartmentalization and its temporal association suggests that neurological symptoms could be linked to the emergence of neurotropic variants.     

Emergent subtype of hepatitis E virus genotype 3 in wild boar in Spain

Wild boar (Sus scrofa) is considered as the main wildlife reservoir of zoonotic hepatitis E virus (HEV) genotypes. The aim of this study was to evaluate the circulation of HEV in free‐ranging wild boar in the Doñana National Park (DNP), Spain. Blood samples were collected from 99 wild boar in the DNP during 2015. Sera were analysed in parallel using indirect ELISA and real‐time RT‐PCR. A total of 57 of the 99 tested animals (57.6%; 95%CI: 47.8%–67.3%) had anti‐HEV antibodies, indicating that this virus is widespread in wild boar in the DNP. HEV RNA was detected in one animal and phylogenetic analysis showed that the sequence isolated belonged to subtype 3r. The results suggest a potential risk of zoonotic transmission of this novel HEV‐3 subtype, which could be of public health concern. Further studies are required to assess the role of wild boar in the epidemiology of HEV‐3r and to determine the infectivity of this emergent HEV subtype in other species, including humans.     

Genetically similar hepatitis E virus strains infect both humans and wild boars in the Barcelona area,Spain, and Sweden

Hepatitis E virus (HEV) is a hepatotropic virus, endemic in Europe where it infects humans and animals, with domestic pigs and wild boars as main reservoirs. The number of HEV‐infected cases with unknown source of infection increases in Europe. There are human HEV strains genetically similar to viruses from domestic pigs, and zoonotic transmission via consumption of uncooked pork meat has been shown. Due to continuous growth of the wild boar populations in Europe, another route may be through direct or indirect contacts with wild boars. In the Collserola Natural Park near Barcelona, Spain, the wild boars have spread into Barcelona city. In Sweden, they are entering into farmlands and villages. To investigate the prevalence of HEV and the risk for zoonotic transmissions, the presence of antibodies against HEV and HEV RNA were analysed in serum and faecal samples from 398 wild boars, 264 from Spain and 134 from Sweden and in sera from 48 Swedish patients with HEV infection without known source of infection. Anti‐HEV was more commonly found in Spanish wild boars (59% vs. 8%; p < 0.0001) while HEV RNA had similar prevalence (20% in Spanish vs. 15% in Swedish wild boars). Seven Swedish and three Spanish wild boars were infected with subtype 3f, and nine Spanish with subtype 3c/i. There were three clades in the phylogenetic tree formed by strains from wild boars and domestic pigs; another four clades were formed by strains from humans and wild boars. One strain from a Spanish wild boar was similar to strains from chronically infected humans. The high prevalence of HEV infections among wild boars and the similarity between wild boar HEV strains and those from humans and domestic pigs indicate that zoonotic transmission from wild boar may be more common than previously anticipated, which may develop into public health concern.     

Serosurvey for Schmallenberg Virus in Alpine Wild Ungulates

Because Schmallenberg virus (SBV) was first reported in domestic ruminants in Northern Italy in February 2012, we conducted a serosurvey to assess the presence of SBV‐specific antibodies in free‐ranging alpine ruminants. The tested serum samples were from chamois (23) and red deer (352) hunted from 2007 to 2013. All of the serum samples collected through September, 2012, tested negative, whereas a single chamois serum and 21 red deer sera taken during the 2012–2013 hunting season tested positive for the presence of SBV antibodies. Because this serosurvey is suggestive of an active SBV circulation in Alpine wildlife, targeted surveillance should be performed on wild ruminants to monitor the spread of the virus and to assess the epidemiological role of wildlife at the interface with domestic animals.     

Prevalence of hepatitis E virus (HEV) infection in various pig farms from Shaanxi Province,China: First detection of HEV RNA in pig semen

The zoonotic transmission of hepatitis E virus (HEV) is mainly mediated by HEV genotypes 3 and 4, with domestic pigs serving as an important reservoir for both genotypes. In China, genotype 4 HEV is the primary prevalent genotype on pig farms. In this study, the prevalence of HEV infection in pig herds of Shaanxi Province was investigated. Serological testing detected anti‐HEV antibody‐positive pigs in five selected cities, with 13 of 17 farms harbouring at least one positive pig (76.47%). Within positive farms, the proportion of positive pigs ranged from 1.6% to 37.5%. Genetic detection analyses of faecal samples revealed that pigs in four cities and on nine of 17 farms were positive for sequences homologous to a partial ORF2‐coding region of HEV (306 bp), as were 6 of 53 bile and 1 of 26 semen samples. Meanwhile, DNA coding for partial HEV ORF1 (1,080 bp) and a longer gene segment coding for partial ORF2 (1,594 bp) were successfully amplified from RNA isolated from pig semen from one HEV‐positive pig. Sequence comparisons of partial ORF2 gene sequences showed that HEV isolates from Shaanxi Province shared the highest identity (81.4%–96.1%) with genotype 4 HEV. Phylogenetic tree analysis grouped these isolates into three subgenotypes (4d, 4h and 4i), with subgenotype 4i the predominant subgenotype. In addition, the HEV isolate from pig semen belonged to subgenotype 4i HEV based on phylogenetic trees constructed both using partial ORF1 and ORF2 gene sequences. In conclusion, HEV infection is endemic on pig farms of Shaanxi Province, China, and 4i is the predominant HEV subgenotype. More important, this is the first study demonstrating detection of HEV RNA in pig semen, suggesting that artificial insemination can transmit HEV in pigs.     

Susceptibility of Pigs to Zoonotic Hepatitis E Virus Genotype 3 Isolated from a Wild Boar

In Europe, zoonotic hepatitis E virus (HEV) genotype 3 strains mainly circulate in humans, swine and wild boar. The aim of this study was to investigate the potential transmission of a wild boar originating HEV strain (WbHEV) to swine by intravenous or oral inoculation and to study the consequences of infection of a WbHEV strain, a WbHEV strain previously passaged in a pig and a swine HEV strain after oral inoculation. Firstly, an intravenous infection was performed for which five piglets were divided into two groups with three pigs inoculated with a WbHEV field strain and two pigs inoculated with a HEV‐negative swine liver homogenate. All pigs were necropsied 8, 9 and 10 days post‐inoculation. Secondly, an oral infection of 56 days was performed on 12 piglets divided into four groups inoculated with a WbHEV strain, a WbHEV strain previously passaged in swine, a swine HEV strain or a HEV‐negative swine liver homogenate. After intravenous inoculation, HEV RNA was detected in serum, bile, liver, spleen, duodenum, jejunum, colon, lung, gastro‐hepatic lymph nodes and faeces in all infected piglets. After oral inoculation, HEV RNA was detected in serum, bile, liver, gastro‐hepatic lymph nodes and faeces. Most of HEV‐inoculated pigs became seropositive at day 15. This study provides experimental evidence of early viral spread throughout the organism after intravenous infection with a WbHEV strain and supports the notion that such a zoonotic strain could be transmitted via the natural faecal–oral route of infection between wild boar and pigs but also between pigs.     

Increased risk of hepatitis E virus infection in workers occupationally exposed to sheep

Hepatitis E virus (HEV) is an enteric RNA virus from the family Hepeviridae with five genotypes (genotypes 1–4 and 7) known to infect humans. HEV infection is known to have a zoonotic swine origin in industrialized countries. The role of pigs and wild boars as major reservoirs for human infection is today well‐established; however, the list of new animal reservoirs is ever‐expanding as new HEV strains are continuously being found in a broad host range. The recent detection of HEV in sheep stools brings concerns on the possibility of HEV transmission from these animals to humans, particularly in those occupationally exposed. The present work investigated the potential occupational risk of HEV infection in shepherds and sheep milk cheesemakers—workers occupationally exposed to ovine (WOEOs; N = 96)—from a region of the Centre of Portugal (‘Serra da Estrela’) based on the differences of anti‐HEV IgG seroprevalence rates between these professionals and the general population (N = 192). The presence of HEV‐specific antibodies in sheep (N = 90) from the same region was also evaluated. The HEV seroprevalence in WOEOs (29.3%) was found to be significantly higher (p = .0198) when compared with population controls (16.1%) which suggests an increased risk for HEV infection in these workers. HEV‐specific antibodies were also found in 16.6% of the studied sheep showing that HEV circulates in these animals. Further studies are needed to confirm the zoonotic potential of sheep HEV.     

Drastic decline of hepatitis E virus detection in domestic pigs after the age of 6 months,Corsica, France

Suidae is an important reservoir of hepatitis E virus (HEV) and a source of transmission to humans (direct contact or via consumption of meat products). Our goal was to characterize the epidemiology of HEV infecting domestic pigs in Corsica Island, a French region hyperendemic for HEV. In Corsica, traditional extensive (or semi‐extensive) outdoor pig farming system is common. Sixteen farms were selected according to location and breeding system. Individual pig faeces samples were collected and qRT‐PCR for detecting HEV RNA was performed. Nucleic acids from HEV‐positive samples were amplified using specific ORF2 genotyping system. The genotype and subtype of the Corsican HEV sequences were determined by phylogenetic analysis. Among the 919 porcine faeces samples tested 9.2% (n = 85) were positive. The presence of viral RNA was correlated with (a) age (>6 months) Adjusted Odd Ratio (AOR) 0.25 [0.068–0.90] p = .032; 3–4 months AOR = 4.94 [2.30–10.62] p = .000043) with the logistic regression model with a random effect at the farm level. Among the 85 positive samples, 83 belonged to genotype 3c and two to genotype 3f. The highest prevalence was observed in the 3–4 months age group and older age (>6 months) was negatively related to HEV infection and this suggests that traditional breeding with a late slaughter age may limit the risk of transmission to humans. A kinetic study of pigs from birth to slaughtering would allow to ensure that the type of traditional breeding reported here is very favourable to the absence of the virus in slaughtered pigs and in pork products.     

Post‐epidemic investigation of Schmallenberg virus in wild ruminants in Slovenia

Schmallenberg virus (SBV) is a vector‐borne virus belonging to the genus Orthobunyavirus within the Bunyaviridae family. SBV emerged in Europe in 2011 and was characterized by epidemics of abortions, stillbirths and congenital malformations in domestic ruminants. The first evidence of SBV infection in Slovenia was from an ELISA‐positive sample from a cow collected in August 2012; clinical manifestations of SBV disease in sheep and cattle were observed in 2013, with SBV RNA detected in samples collected from a total of 28 herds. A potential re‐emergence of SBV in Europe is predicted to occur when population‐level immunity declines. SBV is also capable of infecting several wild ruminant species, although clinical disease has not yet been described in these species. Data on SBV‐positive wild ruminants suggest that these species might be possible sources for the re‐emergence of SBV. The aim of this study was to investigate whether SBV was circulating among wild ruminants in Slovenia and whether these species can act as a virus reservoir. A total of 281 blood and spleen samples from wild ruminants, including roe deer, red deer, chamois and European mouflon, were collected during the 2017–2018 hunting season. Serum samples were tested for antibodies against SBV by ELISA; the overall seroprevalence was 18.1%. Seropositive samples were reported from all over the country in examined animal species from 1 to 15 years of age. Spleen samples from the seropositive animals and serum samples from the seronegative animals were tested for the presence of SBV RNA using real‐time RT‐PCR; all the samples tested negative. Based on the results of the seropositive animals, it was demonstrated that SBV was circulating in wild ruminant populations in Slovenia even after the epidemic, as almost half (23/51) of the seropositive animals were 1 or 2 years old.     

Hepatitis E virus strains infecting Swedish domestic pigs are unique for each pig farm and remain in the farm for at least 2 years

Hepatitis E virus (HEV) genotype 3 (HEV3) is distributed globally and infects both humans and animals, mainly domestic pigs and wild boars, which are the major reservoirs. In this study, the prevalence of HEV among Swedish pigs was investigated by HEV RNA analysis in 363 faecal samples from 3‐month‐old piglets sampled twice (2013 and 2014) in 30 Swedish pig farms. Four different types of farms were investigated; organic, conventional closed (keeping the sow), satellites in a sow pool (conventional farms sharing sows) and conventional non‐closed farms (purchasing gilts). More than two‐thirds (77%) of the farms had HEV‐infected pigs. HEV RNA was found in faeces from 79 pigs (22%). Partial ORF1 could be sequenced in 46 strains. Phylogenetic analysis revealed a unique HEV3 strain for each farm. Strains sampled more than a year apart from the same farm were closely related, indicating that the same HEV strain is present for several years on the farm. Despite that only 4% of the Swedish pig farms were investigated, two farms had strains similar to those from humans, another had strains similar to wild boar HEV. The uniqueness of strains from each farm indicates a possibility to identify a source of infection down to farm level. This knowledge may be used by the farms to investigate the effectiveness of good hygiene routines to reduce the amount of HEV and thus the infection risk in the farm, and for Swedish public health authorities to identify cases of HEV transmissions from consumption of uncooked pork.     

Absence of Hepatitis E virus circulation in wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis) in Mediterranean ecosystems in Spain

In recent decades, cases of autochthonous hepatitis E (HE) have sharply increased in European countries where foodborne transmission is considered the main route of HE virus (HEV) transmission. Although rabbits are considered the main reservoir of the zoonotic HEV‐3ra subtype, information on the role of wild lagomorphs in the epidemiology of HEV remains scarce. The aim of this study therefore was to assess the circulation of HEV in European wild rabbits (Oryctolagus cuniculus) and Iberian hares (Lepus granatensis), the most important lagomorph species in Spanish Mediterranean ecosystems. Liver samples from 372 wild rabbits and 78 Iberian hares were analysed using a broad‐spectrum RT‐PCR that detects HEV genotypes 1–8. None of the 450 lagomorphs tested were positive for HEV infection. To the best of our knowledge, this is the first study to assess HEV circulation in wild rabbits in Spain and the first to evaluate HEV infection in Iberian hares. Our results indicate absence of HEV circulation in wild rabbits and Iberian hares in southern Spain during the study period, which suggests that the risk of transmission of HEV from wild lagomorphs to other species, including humans, is low.