Detection of antibodies to the hepatitis E virus in domestic reindeer (Rangifer tarandus) in the Republic of Sakha (Yakutia).

INTRODUCTION: Although domestic pigs and wild boars are the main reservoir of zoonotic hepatitis E virus (HEV) genotypes in temperate countries, the presence of antibodies to HEV (anti-HEV) in the indigenous population of circumpolar territories, i.e. outside the habitat of wild and domestic pigs, indicates the presence of an alternative reservoir of the virus. Reindeer (Rangifer tarandus) may be a potential reservoir for HEV in the polar regions. The purpose of the study was to determine the prevalence of anti-HEV among domestic reindeer in the Republic of Sakha (Yakutia). MATERIALS AND METHODS: Sera from 497 domestic reindeer from the Oymyakon (n = 425) and Ust-Yansky districts (n = 72) of the Republic of Sakha (Yakutia) were tested for anti-HEV. A commercial ELISA kit DS-ELISA-ANTI-HEV-G (Diagnostic Systems-Stolitsa LLC, Russia) was used for detection of anti-HEV IgG, but a rabbit polyclonal antibody against deer IgG labeled with horseradish peroxidase (KPL, USA) at a dilution of 1 : 100 in phosphate-buffered saline were used instead of the human specific conjugate from the kit. RESULTS: The average detection rate of anti-HEV in reindeer sera was 15.5% (95% CI: 12.6-19.0%). The detection rate of anti-HEV significantly increased with age, from 3.5% (95% CI: 1.1-9.0%) in calves aged 3-6 months to 25.0% (95% CI: 1.6 -36.5%) in deer aged 2-4 years (p < 0.0001). From this age group, anti-HEV detection rates reached a plateau, not differing significantly between older age groups (p > 0.05). The average anti-HEV detection rate among reindeer 2 years of age and older was 19.0% (95% CI: 15.3-23.4%). There were no statistically significant differences in the frequency of anti-HEV detection between female and male reindeer, both among adult animals and among calves. CONCLUSION: The observed anti-HEV detection rates among domestic reindeer in the Republic of Sakha (Yakutia) indicate that infection caused by HEV or an antigenically similar virus is common in these animals. The dynamics of antibody accumulation in the reindeer population indicates that infection apparently occurs during the first two years of life.

Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus.

The dicistrovirus intergenic region internal ribosome entry site (IGR IRES) uses an unprecedented, streamlined mechanism whereby the IRES adopts a triple-pseudoknot (PK) structure to directly bind to the conserved core of the ribosome and drive translation from a non-AUG codon. The origin of this IRES mechanism is not known. Previously, a partial fragment of a divergent dicistrovirus RNA genome, named ancient Northwest territories cripavirus (aNCV), was extracted from 700-year-old caribou feces trapped in a subarctic ice patch. The aNCV IGR sequence adopts a secondary structure similar to contemporary IGR IRES structures, however, there are subtle differences including 105 nucleotides upstream of the IRES of unknown function. Using filter binding assays, we showed that the aNCV IRES could bind to purified ribosomes, and toeprinting analysis pinpointed the start site at a GCU alanine codon adjacent to PKI. Using a bicistronic reporter RNA, the aNCV IGR can direct translation in vitro in a PKI-dependent manner. Lastly, a chimeric infectious clone swapping in the aNCV IRES supported translation and virus infection. The characterization and resurrection of a functional IGR IRES from a divergent 700-year-old virus provides a historical framework for the importance of this viral translational mechanism.

Seroprevalence of Hepatitis E Virus in Moose (Alces alces), Reindeer (Rangifer tarandus), Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Muskoxen (Ovibos moschatus) from Norway.

Hepatitis E virus (HEV), a major cause of viral hepatitis worldwide, is considered an emerging foodborne zoonosis in Europe. Pigs (Sus scrofa domestica) and wild boars (S. scrofa) are recognized as important HEV reservoirs. Additionally, HEV infection and exposure have been described in cervids. In Norway, HEV has been identified in pigs and humans; however, little is known regarding its presence in wild ungulates in the country. We used a species-independent double-antigen sandwich ELISA to detect antibodies against HEV in the sera of 715 wild ungulates from Norway, including 164 moose (Alces alces), 186 wild Eurasian tundra reindeer (Rangifer tarandus tarandus), 177 red deer (Cervus elaphus), 86 European roe deer (Capreolus capreolus), and 102 muskoxen (Ovibos moschatus). The overall seroprevalence was 12.3% (88/715). Wild reindeer had the highest seropositivity (23.1%, 43/186), followed by moose (19.5%, 32/164), muskoxen (5.9%, 6/102), and red deer (4%, 7/177). All roe deer were negative. According to our results, HEV is circulating in wild ungulates in Norway. The high seroprevalence observed in wild reindeer and moose indicates that these species may be potential reservoirs of HEV. To the authors' knowledge, this is the first report of HEV exposure in reindeer from Europe and in muskoxen worldwide.

Permafrost dynamics and the risk of anthrax transmission: a modelling study.

A recent outbreak of anthrax disease, severely affecting reindeer herds in Siberia, has been reportedly associated to the presence of infected carcasses or spores released from the active layer over permafrost, which is thawing and thickening at increasing rates, thus underlying the re-emerging nature of this pathogen in the Arctic region because of warming temperatures. Anthrax is a global zoonotic and epizootic disease, with a high case-fatality ratio in infected animals. Its transmission is mediated by environmental contamination through highly resistant spores which can persist in the soil for several decades. Here we develop and analyze a new epidemiological model for anthrax transmission that is specifically tailored to the Arctic environmental conditions. The model describes transmission dynamics including also herding practices (e.g. seasonal grazing) and the role of the active layer over permafrost acting as a long-term storage of spores that could be viable for disease transmission during thawing periods. Model dynamics are investigated through linear stability analysis, Floquet theory for periodically forced systems, and a series of simulations with realistic forcings. Results show how the temporal variability of grazing and active layer thawing may influence the dynamics of anthrax disease and, specifically, favor sustained pathogen transmission. Particularly warm years, favoring deep active layers, are shown to be associated with an increase risk of anthrax outbreaks, and may also foster infections in the following years. Our results enable preliminary insights into measures (e.g. changes in herding practice) that may be adopted to decrease the risk of infection and lay the basis to possibly establish optimal procedures for preventing transmission; furthermore, they elicit the need of further investigations and observation campaigns focused on anthrax dynamics in the Arctic environment.

Ocular Histopathological Findings in Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus) with Infectious Keratoconjunctivitis after Experimental Inoculation with Cervid Herpesvirus 2.

Infectious keratoconjunctivitis (IKC) is a common transmissible ocular disease in semi-domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus). In large outbreaks, IKC may affect tens of animals in a herd, with the most severe cases often requiring euthanasia due to the destruction of the affected eyes and permanent blindness. An experimental inoculation with cervid herpesvirus 2 (CvHV2), alone or in combination with Moraxella bovoculi, demonstrated that CvHV2 has the ability to cause clinical signs of IKC in previously unexposed reindeer. Tissues collected from upper and lower eyelids, lacrimal gland and cornea, were processed for light and transmission electron microscopy. Histopathological analysis of the eyes inoculated with CvHV2 showed widespread and severe pathological findings. Mucosal tissues from these eyes showed fibrinous and purulent exudates, hyperemia, hemorrhages, necrosis, vascular thrombosis, vascular necrosis, infiltration of mononuclear cells and neutrophils, and lymphoid follicle reaction, which matches the described histopathology of IKC in reindeer. Characteristic alpha-herpesvirus particles matching the size and morphology of CvHV2 were identified by transmission electron microscopy in the conjunctival tissue. The quantification of viral particles by qPCR revealed high copy numbers of viral DNA in all CvHV2 inoculated eyes, but also in the non-inoculated eyes of the same animals. The histopathology of eye tissues obtained from the CvHV2 inoculated reindeer and the lack of inflammation from bacterial infection, together with the detection of CvHV2 DNA in swabs from the inoculated and non-inoculated eyes of the same animals, verified that CvHV2 was the primary cause of the observed histopathological changes.

Gammaherpesvirus in Cervid Species from Norway: Characterization of a New Virus in Wild and Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus).

Gammaherpesvirus infections have been described in cervids worldwide, mainly the genera Macavirus or Rhadinovirus. However, little is known about the gammaherpesviruses species infecting cervids in Norway and Fennoscandia. Blood samples from semi-domesticated (n = 39) and wild (n = 35) Eurasian tundra reindeer (Rangifer tarandus tarandus), moose (Alces alces, n = 51), and red deer (Cervus elaphus, n = 41) were tested using a panherpesvirus DNA polymerase (DPOL) PCR. DPOL-PCR-positive samples were subsequently tested for the presence of glycoprotein B (gB) gene. The viral DPOL gene was amplified in 28.2% (11/39) of the semi-domesticated reindeer and in 48.6% (17/35) of the wild reindeer. All moose and red deer tested negative. Additionally, gB gene was amplified in 4 of 11 semi-domesticated and 15 of 17 wild Eurasian reindeer DPOL-PCR-positive samples. All the obtained DPOL and gB sequences were highly similar among them, and corresponded to a novel gammaherpesvirus species, tentatively named Rangiferine gammaherpesvirus 1, that seemed to belong to a genus different from Macavirus and Rhadinovirus. This is the first report of a likely host-specific gammaherpesvirus in semi-domesticated reindeer, an economic and cultural important animal, and in wild tundra reindeer, the lastpopulation in Europe. Future studies are required to clarify the potential impact of this gammaherpesvirus on reindeer health.

Pestivirus Infections in Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus): A Retrospective Cross-Sectional Serological Study in Finnmark County, Norway.

Members of the Pestivirus genus (family Flaviviridae) cause severe and economically important diseases in livestock. Serological studies have revealed the presence of pestiviruses in different cervid species, including wild and semi-domesticated Eurasian tundra reindeer. In this retrospective study, serum samples collected between 2006 and 2008 from 3339 semi-domesticated Eurasian reindeer from Finnmark County, Norway, were tested for anti-pestivirus antibodies using an enzyme linked immunosorbent assay (ELISA) and a subset of these by virus neutralization test (VNT). A seroprevalence of 12.5% was found, varying from 0% to 45% among different herding districts, and 20% in western Finnmark, as compared to 1.7% in eastern Finnmark. Seroprevalence increased with age. Pestivirus-specific RNA was not detected in any of the 225 serum samples tested by real-time RT-PCR. Based on VNT results, using a panel of one bovine viral diarrhea virus (BVDV) strain and two border disease virus (BDV) strains, the virus is most likely a reindeer-specific pestivirus closely related to BDV. A characterization of the causative virus and its pathogenic impact on reindeer populations, as well as its potential to infect other domestic and wild ruminants, should be further investigated.

Orf virus infection in Alaskan mountain goats, Dall's sheep, muskoxen, caribou and Sitka black-tailed deer.

BACKGROUND: The zoonotic Orf virus (ORFV; genus Parapoxvirus, Poxviridae family) occurs worldwide and is transmitted between sheep and goats, wildlife and man. Archived tissue samples from 16 Alaskan wildlife cases, representing mountain goat (Oreamnos americanus, n = 8), Dall's sheep (Ovis dalli dalli, n = 3), muskox (Ovibos moschatus, n = 3), Sitka black-tailed deer (Odocoileus hemionus sitkensis, n = 1) and caribou (Rangifer tarandus granti, n = 1), were analyzed. RESULTS: Clinical signs and pathology were most severe in mountain goats, affecting most mucocutaneous regions, including palpebrae, nares, lips, anus, prepuce or vulva, as well as coronary bands. The proliferative masses were solid and nodular, covered by dark friable crusts. For Dall's sheep lambs and juveniles, the gross lesions were similar to those of mountain goats, but not as extensive. The muskoxen displayed ulcerative lesions on the legs. The caribou had two ulcerative lesions on the upper lip, as well as lesions on the distal part of the legs, around the main and dew claws. A large hairless spherical mass, with the characteristics of a fibroma, was sampled from a Sitka black-tailed deer, which did not show proliferative lesions typical of an ORFV infection. Polymerase chain reaction analyses for B2L, GIF, vIL-10 and ATI demonstrated ORFV specific DNA in all cases. Sequences from Dall's sheep formed a separate cluster, comparable to ORFV from domestic sheep. Sequences from the other species were different from the Dall's sheep sequences, but almost identical to each other. CONCLUSIONS: This is the first major investigation of parapoxvirus infections in large Alaskan game species, and the first report of parapoxvirus infection in caribou and Sitka black-tailed deer. This study shows that most of the wild ruminant species in Alaska and from most parts of Alaska, can carry and be affected by ORFV. These findings call for attention to transmission of ORFV from wildlife to livestock and to hunters, subsistence harvesters, and wildlife biologists.

Infectious keratoconjunctivitis in semi-domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus): microbiological study of clinically affected and unaffected animals with special reference to cervid herpesvirus 2.

BACKGROUND: Infectious keratoconjunctivitis (IKC) is one of the most common ocular diseases in ruminants worldwide. In addition to keratitis and conjunctivitis, animals with IKC can develop uveitis, corneal ulcer, and in severe cases, blindness. The bacteria Moraxella spp. has been described as the primary causative agent of infectious bovine keratoconjunctivitis (IBK) in cattle (Bos taurus), while Chlamydia spp. and Mycoplasma conjunctivae are considered the main causative agents of IKC in sheep (Ovis aries). Previous studies indicated cervid herpesvirus 2 (CvHV2) as the primary causative agent of IKC in semi-domesticated reindeer (Rangifer tarandus tarandus). The aim of the study was to investigate the presence and prevalence of potential pathogens for IKC in reindeer, and compare the ocular microbiota of animals with IKC, with apparently healthy animals. RESULTS: Semi-domesticated reindeer (n = 341), with (n = 108) or without (n = 113) ocular clinical signs, or with no information on clinical status (n = 120), were sampled in Norway, Sweden and Finland in 2010-2014. Seroprevalence was 37.4% for alphaherpesvirus (95/254), 3.8% for gammaherpesvirus (8/211) and 7.1% for pestivirus (15/211) (ELISA). PCR analyses of conjunctival swab samples revealed a prevalence of 28.5% for CvHV2 (57/200), 11.9% for Chlamydiaceae (16/135) and 1.0% for M. conjunctivae (2/197). Bacteriological cultivation of 202 conjunctival swab samples revealed bacterial growth from 75.2% of the samples, with Moraxella spp. being isolated from 21.6% (11/51) of the animals with and 5.6% (5/84) without ocular clinical signs. A significant association (p < 0.001) existed between the presence of clinical signs of IKC and CvHV2 DNA in the affected eyes, an association that was not present for other microorganisms. CONCLUSIONS: These results support the hypothesis that CvHV2 is the primary agent of IKC in semi-domesticated reindeer in Fennoscandia, with Moraxella bovoculi being a secondary candidate, since it was isolated in two different outbreaks of IKC. Further studies should be carried out to better understand the infection biology and the pathogenesis of IKC in reindeer.

Metagenomic survey for viruses in Western Arctic caribou, Alaska, through iterative assembly of taxonomic units.

Pathogen surveillance in animals does not provide a sufficient level of vigilance because it is generally confined to surveillance of pathogens with known economic impact in domestic animals and practically nonexistent in wildlife species. As most (re-)emerging viral infections originate from animal sources, it is important to obtain insight into viral pathogens present in the wildlife reservoir from a public health perspective. When monitoring living, free-ranging wildlife for viruses, sample collection can be challenging and availability of nucleic acids isolated from samples is often limited. The development of viral metagenomics platforms allows a more comprehensive inventory of viruses present in wildlife. We report a metagenomic viral survey of the Western Arctic herd of barren ground caribou (Rangifer tarandus granti) in Alaska, USA. The presence of mammalian viruses in eye and nose swabs of 39 free-ranging caribou was investigated by random amplification combined with a metagenomic analysis approach that applied exhaustive iterative assembly of sequencing results to define taxonomic units of each metagenome. Through homology search methods we identified the presence of several mammalian viruses, including different papillomaviruses, a novel parvovirus, polyomavirus, and a virus that potentially represents a member of a novel genus in the family Coronaviridae.

Identification and characterization of two novel viruses in ocular infections in reindeer.

A thorough understanding of virus diversity in wildlife provides epidemiological baseline information about pathogens. In this study, eye swab samples were obtained from semi-domesticated reindeer (Rangifertarandus tarandus) in Norway during an outbreak of infectious eye disease, possibly a very early stage of infectious keratoconjunctivitis (IKC). Large scale molecular virus screening, based on host nucleic acid depletion, sequence-independent amplification and next-generation sequencing of partially purified viral nucleic acid, revealed the presence of a new papillomavirus in 2 out of 8 eye swab samples and a new betaherpesvirus in 3 out of 8 eye swab samples collected from animals with clinical signs and not in similar samples in 9 animals without clinical signs. Whether either virus was responsible for causing the clinical signs or in any respect was associated to the disease condition remains to be determined.

Gammaherpesvirus infection in semidomesticated reindeer (Rangifer tarandus tarandus): a cross-sectional, serologic study in northern Norway.

Malignant catarrhal fever (MCF) is caused by a group of gammaherpesviruses that primarily affect domestic and wild ruminants. Using competitive-inhibition enzyme-linked immunosorbent assay, we screened 3,339 apparently healthy, semidomesticated reindeer (Rangifer tarandus tarandus) from Finnmark County, Norway, sampled during slaughter. The overall antibody prevalence was 3.5% and varied among reindeer herding districts in Finnmark (0-6.7%), the largest reindeer herding region in Norway. The risk of exposure to gammaherpesvirus (i.e., seroconversion) was significantly higher for adult reindeer than it was for calves ≤1 yr, for reindeer in east Finnmark (3.8%) compared with west Finnmark (3.3%), and with increasing population density. No evidence of disease associated with this virus was detected in reindeer sampled for this study, but because samples were collected at slaughterhouses, one cannot discard the possibility of these events happening in the field. The low antibody prevalence could indicate occasional infection of reindeer with another ruminant gammaherpesvirus or the presence of a yet-unknown, specific, low-pathogenic reindeer gammaherpesvirus. Further studies should aim at characterizing the virus circulating in reindeer and address the potential clinical impact of this virus.

Public health implications of an outbreak of rabies in arctic foxes and reindeer in the Svalbard archipelago, Norway, September 2011.

Between 16 September and 5 October 2011 rabies was diagnosed in two arctic foxes and eight reindeer in the Svalbard archipelago, in Norway. This outbreak occurs at the end of the reindeer hunting season and poses an increased risk to many people that were involved in the hunt. As of 28 September 2011, 280 people had received post-exposure prophylaxis. No human cases of rabies have occurred.

Cervid herpesvirus 2 infection in reindeer: a review.

Herpesviruses of the genus Varicellovirus are known to infect and cause disease in a variety of ruminant species, but the impact of cervid herpesvirus 2 (CvHV2) in reindeer (Rangifer tarandus) is mostly unknown. Reindeer is a circum-polar species with a total estimated number of more than 5 million animals. Mortality may reach high values, as in northern Norway, especially in calves (37%; 2005-2006), and disease can potentially account for some of this mortality. CvHV2 has been isolated during a natural outbreak of infectious keratoconjunctivitis, indicating an etiologal link. Serological screening has shown that CvHV2 infection is prevalent in Northern Norway and experimental infection studies have demonstrated that viremia, latency and vertical transmission occur for CvHV2. The present review aims at summarizing current knowledge on the epidemiology, pathogenesis and molecular virology of CvHV2.

The genome of pseudocowpoxvirus: comparison of a reindeer isolate and a reference strain.

Parapoxviruses (PPV), of the family Poxviridae, cause a pustular cutaneous disease in sheep and goats (orf virus, ORFV) and cattle (pseudocowpoxvirus, PCPV and bovine papular stomatitis virus, BPSV). Here, we present the first genomic sequence of a reference strain of PCPV (VR634) along with the genomic sequence of a PPV (F00.120R) isolated in Finland from reindeer (Rangifer tarandus tarandus). The F00.120R and VR634 genomes are 135 and 145 kb in length and contain 131 and 134 putative genes, respectively, with their genome organization being similar to that of other PPVs. The predicted proteins of F00.120R and VR634 have an average amino acid sequence identity of over 95%, whereas they share only 88 and 73% amino acid identity with the ORFV and BPSV proteomes, respectively. The most notable differences were found near the genome termini. F00.120R lacks six and VR634 lacks three genes seen near the right terminus of other PPVs. Four genes at the left end of F00.120R and one in the middle of both genomes appear to be fragmented paralogues of other genes within the genome. VR634 has larger than expected inverted terminal repeats possibly as a result of genomic rearrangements. The high G+C content (64%) of these two viruses along with amino acid sequence comparisons and whole genome phylogenetic analyses confirm the classification of PCPV as a separate species within the genus Parapoxvirus and verify that the virus responsible for an outbreak of contagious stomatitis in reindeer over the winter of 1999-2000 can be classified as PCPV.

Experimental infection of reindeer with cervid herpesvirus 2.

Cervid herpesvirus 2 (CvHV2) has been isolated from reindeer (Rangifer tarandus tarandus), and serological data indicate that in reindeer this virus is endemic in Fennoscandia, Alaska, Canada, and Greenland. CvHV2 has been described as a cause of subclinical genital infections in reindeer, but little information on primary infections exists. In this study, six seronegative and presumably pregnant reindeer were allocated to one of two groups. Two animals were inoculated with CvHV2 intratracheally, and two animals intravaginally, with one control animal in each group receiving sterile water. Mild hyperthermia and serous discharges from the vagina and nose were observed. No abortions were recorded, but one calf died shortly after birth. Inoculated animals seroconverted and had neutralizing antibodies after days 7 to 10 postinfection. CvHV2 was detected by PCR in nasal and vaginal swabs from animals in both groups but could be isolated only from nasal swabs in the respiratory group and from vaginal swabs in the genital group. CvHV2 was detected by PCR in various organs and tissues postmortem. In control animals, the virus could not be isolated in spite of PCR-positive nasal and vaginal swab samples and some degree of positive immunostaining. One of the animals that were inoculated intratracheally developed a hemorrhagic, necrotizing bronchopneumonia, which was CvHV2 positive by PCR and immunohistochemistry. We conclude that CvHV2 can cause systemic infection, that both genital and respiratory inoculations can lead to virus shedding, and that the virus can infect the fetus in utero.

Cervid herpesvirus 2, the primary agent in an outbreak of infectious keratoconjunctivitis in semidomesticated reindeer.

An outbreak of infectious keratoconjunctivitis (IKC) occurred in semidomesticated reindeer (Rangifer tarandus tarandus) in Troms County, Norway, in February 2009. Twenty-eight animals with clinical symptoms and 12 apparently healthy animals were investigated. They ranged in age from calves of the year to 4-year-old animals (mean, 1.9 years; standard deviation, +/-0.9). The seroprevalence of antibodies against cervid herpesvirus 2 (CvHV2) was 86% in animals with IKC and 42% in unaffected animals. For the 28 clinically affected animals, CvHV2 was detected by PCR in swabs obtained from the eye (82%), nose (64%), and vagina (24%), and CvHV2 was isolated from eye swabs from 8 animals. Virus was not isolated from clinically unaffected animals but was detected by PCR in eye swab samples from five of them. The viral activity, assessed by the ability to cause a cytopathic effect in cell culture, increased with the severity of clinical symptoms, but in severe clinical cases, virus was absent and secondary bacterial infections were dominant. Moraxella sp. isolates were obtained from seven animals, and those from two animals were identified as Moraxella bovoculi. Staphylococcus aureus, Streptococcus sp., and Arcanobacterium pyogenes were also isolated. It is concluded that CvHV2, which is endemic in reindeer, can cause IKC, probably most commonly as a primary infection of calves. This can be a very painful and devastating disease of economic importance for reindeer herders. This is the first report of CvHV2 as the primary agent of IKC in reindeer. This is also the first isolation of this virus in reindeer under natural herding conditions.

Cervid herpesvirus 2 causes respiratory and fetal infections in semidomesticated reindeer.

Members of the viral subfamily Alphaherpesvirinae establish latency from which they can be reactivated. Bovine herpesvirus 1 causes infectious bovine rhinotracheitis and infectious pustular vulvovaginitis in cattle, as well as abortion and weak calves. Serological evidence of alphaherpesvirus infection has been reported for wild and semidomesticated reindeer (Rangifer tarandus tarandus) in Norway. To address the possibility that reindeer alphaherpesvirus (cervid herpesvirus 2 [CvHV-2]) infection might affect the respiratory system and in part explain the relatively high mortality of reindeer calves during their first year, tissue samples were obtained from reindeer and reindeer fetuses at slaughterhouses in Finnmark County, Norway. A nested pan-alphaherpesvirus PCR amplification targeting the highly conserved UL27 gene (encoding glycoprotein B) was used. Sequencing of amplicons revealed the presence of CvHV-2 DNA. The detection of CvHV-2 DNA in trigeminal ganglia (27 of 143 samples), nasal swabs (5 of 75 samples), and fetal tissues (12 of 48 samples) indicates that CvHV-2 infection is endemic in this reindeer population. Moreover, the virus is transmitted horizontally by the respiratory route, establishing latency in the trigeminal ganglion, and vertically to the fetus through the placenta. Further studies should focus on the reproductive impact of CvHV-2 infection in reindeer.

Evaluation of three commercial bovine ELISA kits for detection of antibodies against Alphaherpesviruses in reindeer (Rangifer tarandus tarandus).

BACKGROUND: The genus Varicellovirus (family Herpesviridae subfamily Alphaherpesvirinae) includes a group of viruses genetically and antigenically related to bovine herpesvirus 1 (BoHV-1) among which cervid herpesvirus 2 (CvHV-2) can be of importance in reindeer. These viruses are known to be responsible for different diseases in both wild and domestic animals. Reindeer are a keystone in the indigenous Saami culture and previous studies have reported the presence of antibodies against alphaherpesviruses in semi-domesticated reindeer in northern Norway. Mortality rates, especially in calves, can be very high in some herds and the abortion potential of alphaherpesvirus in reindeer, unlike in bovines, remains unknown. ELISA kits are the most used screening method in domestic ruminants and given the close genetic relationship between viruses within this genus, it might be possible to use such kits to screen cervids for different alphaherpesviruses. We have compared three different commercial ELISA kits in order to validate its use for reindeer and CvHV-2. METHODS: Three commercial bovine ELISA kits (A, B and C), using either indirect (A) or blocking (B and C) ELISA techniques to detect antibodies against BoHV-1 were tested with sera from 154 reindeer in order to detect antibodies against CvHV-2. A Spearman's rank-based coefficient of correlation (rho) was calculated. A dilution trial was performed for all kits. A virus neutralization test using both BoHV-1 and CvHV-2 was carried out. RESULTS: Seroprevalence was almost the same with all kits (40-41%). Despite a similar qualitative score, quantitatively kits classified samples differently and a strong correlation was only identified between Kits B and C. Blocking kits performed better in both repeatability and in the dilution trial. The virus neutralization results confirmed the ELISA results to a very high degree. Neutralizing titres ranged from 1:2 to 1:256 and from 0 to 1:16 against CvHV-2 and BoHV-1 respectively. CONCLUSION: Results show that the genetic and antigenic similarity between BoHV-1 and CvHV-2 enables the use of a bovine gB blocking ELISA kit to screen reindeer. The use of an ELISA kit is both cheaper and time saving, allowing screening of large populations. This study revealed a high number of positive animals against CvHV-2 and its impact and distribution in the general population should be further evaluated.

Serological evidence of hepatitis E virus infection in an indigenous North American population.

BACKGROUND: Hepatitis E virus (HEV) infections are thought to be uncommon in North America. Recently, HEV transmission has been reported following the consumption of deer meat. Because deer are closely related to caribou and caribou meat is a staple of the Canadian Inuit and the American Eskimo diet, the present study explored the seroprevalence of HEV infection in an isolated Canadian Inuit community. METHODS: Stored sera were thawed and tested for immunoglobulin (Ig) G and IgM anti-HEV by ELISA, and tested for HEV-RNA by reverse transcriptase polymerase chain reaction. RESULTS: The study consisted of 393 sera (representing approximately 50% of the community's inhabitants). Eleven samples (3%) were IgG anti-HEV-positive. Their mean age was 29+/-8 years and three were male. Two of 11 (18%) were also IgM anti-HEV-positive. All IgG anti-HEV-positive individuals were HEV-RNA-negative. Liver biochemistry was normal in all. Seven of 11 (64%) were also positive for anti-hepatitis A virus, five (46%) were hepatitis B virus seropositive and none (0%) were positive for anti-hepatitis C virus. There were no associations between infections with HEV and other hepatropic viruses. Serological testing was negative for HEV infection in 25 caribou from an adjacent region. CONCLUSION: The results of the present study showed that serological evidence of HEV infection was present in 3% of the observed Canadian Inuit population; the presence of IgM anti-HEV suggested recent infection and HEV did not appear to coinfect with other common hepatotropic viruses. The source of HEV infection in the population remains unclear. These findings are interesting but preliminary. Additional data are required to determine whether HEV infections are responsible for otherwise unexplained acute hepatitis in the Canadian Inuit population and visitors returning from northern North American communities.