Hepatitis E Virus Antibody Prevalence in Wildlife in Poland

Hepatitis E is an important public health problem mostly in developing but occasionally also in industrialized countries. Domestic and wildlife animals are considered reservoirs of the hepatitis E virus (HEV). Since no information on the prevalence of autochthonous HEV infections in human and animal in Poland is available, the aim of the study was to investigate the HEV seroprevalence of different wildlife species as potential virus reservoirs in the country. No HEV antibodies were found in any of the sera collected from the red deer (Cervus elaphus), European bison (Bison bonasus), roe deer (Capreolus capreolus), elk (Alces alces), fallow deer (Dama dama), sika deer (Cervus nippon), Tatra chamois (Rupicapra rupicapra tatrica) or brown bear (Ursus arctos). HEV‐specific antibodies were detected in 44.4% (95% CI 38.3–50.7) serum samples originated only from wild boars. The percentage of seropositive wild boars differed significantly between the provinces and was positively correlated with the wild boar density and rurality of the area. This study showed that HEV circulates among wild boar population in Poland, and this species should be considered as an important reservoir of the virus.     

Immunoglobulin G Class Identification from Wild Ungulates by Cross‐reactivity with Antisera to Domestic Animals

Seven species of Spanish ungulates were tested for the presence of homologous immunoglobulin G (IgG) with a gel‐diffusion test using bovine, ovine, caprine and porcine IgG antisera. Homologous ovine and caprine IgG were detected in sera from chamois (Rupicapra pyrenaica), Spanish ibex (Capra pyrenaica hispanica), mouflon (Ovis orientalis musimon), red deer (Cervus elaphus), fallow deer (Dama dama) and roe deer (Capreolus capreolus). Homologous porcine IgG was detected in wild boar (Sus scrofa) serum. Immunoelectrophoretic assays were performed to compare the electrophoretic mobility of IgG from domestic and wild species.     

Babesia spp. in European wild ruminant species: parasite diversity and risk factors for infection

Babesia are tick-borne parasites that are increasingly considered as a threat to animal and public health. We aimed to assess the role of European free-ranging wild ruminants as maintenance mammalian hosts for Babesia species and to determine risk factors for infection. EDTA blood was collected from 222 roe deer (Capreolus c. capreolus), 231 red deer (Cervus e. elaphus), 267 Alpine chamois (Rupicapra r. rupicapra) and 264 Alpine ibex (Capra i. ibex) from all over Switzerland and analysed by PCR with pan-Babesia primers targeting the 18S rRNA gene, primers specific for B. capreoli and Babesia sp. EU1, and by sequencing. Babesia species, including B. divergens, B. capreoli, Babesia sp. EU1, Babesia sp. CH1 and B. motasi, were detected in 10.7% of all samples. Five individuals were co-infected with two Babesia species. Infection with specific Babesia varied widely between host species. Cervidae were significantly more infected with Babesia spp. than Caprinae. Babesia capreoli and Babesia sp. EU1 were mostly found in roe deer (prevalences 17.1% and 7.7%, respectively) and B. divergens and Babesia sp. CH1 only in red deer. Factors significantly associated with infection were low altitude and young age. Identification of Babesia sp. CH1 in red deer, co-infection with multiple Babesia species and infection of wild Caprinae with B. motasi and Babesia sp. EU1 are novel findings. We propose wild Caprinae as spillover or accidental hosts for Babesia species but wild Cervidae as mammalian reservoir hosts for B. capreoli, possibly Babesia sp. EU1 and Babesia sp. CH1, whereas their role regarding B. divergens is more elusive.     

High incidence of single nucleotide polymorphisms in the prion protein gene of native Brazilian Caracu cattle

Different alleles of the human and ovine prion protein gene correlate with a varying susceptibility to transmissible spongiform encephalopathies. However, the pathogenic implications of specific polymorphisms in the bovine prion protein gene (PRNP) are only poorly understood. Previous studies on the bovine PRNP gene investigated common European and North American cattle breeds. As a consequence of decades of intensive breeding for specific traits, these modern breeds represent only a small fraction of the bovine gene pool. In this study, we analysed PRNP polymorphisms in the native Brazilian Caracu breed, which developed in geographical isolation since the 16th century. A total of 10 single nucleotide polymorphisms (SNPs) were discovered in the coding region of the Caracu PRNP gene. Eight of the SNPs occurred at high frequencies in Caracu cattle (variant allele frequencies = 0.10–0.76), but were absent or only rarely observed in European and North American breeds. One of the Caracu SNPs was associated with an amino acid exchange from serine to asparagine (f = 0.17). This SNP was not detected in Holstein–Friesian, Simmental and German Gelbvieh and was only rarely detected in beef cattle (f = 0.01). We found 17 haplotypes for PRNP in the Caracu breed.     

Management of Wild Ungulate Populations in Italy: Captive-Breeding,Hybridisation and Genetic Consequences of Translocations

Captive-reproduced stocks of some species of ungulates (Artiodactyla), and particularly the red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus) and the wildboar (Sus scrofa) are more or less extensively translocated in Italy, mainly for local reintroductions or restocking of exploited wild populations. However, captive breeding often involves the reproduction of non-indigenous individuals or the production of artificial hybrids. Consequently, translocations of captive-reproduced ungulates are of concern for the conservation of indigenous populations and gene pools. The impact of translocations should be evaluated within the background of the growing knowledge on population genetic and phylogeographic structure of ungulates. Molecular genetic markers are being used to map geographic genetic diversity, and reconstruct the phylogeographic history of natural populations (i.e., in the roe deer). Molecular makers are also used to detect the consequences of domestication and identify hybrids between wild and domesticated populations (i.e., in the wildboar), or to detect inter-specific hybridisation (i.e., between the red deer and wapiti). Hybridisation of wild and domestic pigs, and diffusion of hybrids in nature is widespread in Italy. Admixture of indigenous and non-indigenous roe deer stocks is also widespread. Therefore, conservation and management of indigenous ungulates calls for careful evaluation of captive-reproduced stocks.     

Seroprevalence of pestivirus in four species of alpine wild ungulates in the High Valley of Susa, Italy

Wildlife, once infected, can serve as a reservoir of infectious diseases that form a constant threat to domestic livestock. To make control and eradication programs successful in the long-term, presence of pestivirus in wildlife populations should be monitored. The goal of this study was to investigate seroprevalence of pestivirus in four alpine wild ungulates in the High Valley of Susa, north-west Italy. Species studied were: red deer (Cervus elaphus), roe deer (Capreolus capreolus), wild boar (Sus scrofa) and chamois (Rupicapra rupicapra). A further goal was using virus neutralisation tests (VNT) for four strains of pestivirus in chamois and wild boar. Three hundred and seventy-five serum samples collected during the hunting season of 1999 were tested for pestivirus specific antibodies. Positive sera of chamois and wild boar were subsequently tested in a VNT with four major subtypes of pestivirus, and virus isolation was performed. No antibodies were found in the 73 samples of roe deer, while 7 (12.5%), 8 (5.9%) and 28 (25.5%) of 56, 136 and 110 samples of wild boar, red deer and chamois were ELISA-positive, respectively. Different ranges of titers were found in the VNT and no pestivirus was isolated in the ELISA-positive wild boar and chamois samples. Several possibilities, which might explain the high seroprevalence in chamois are discussed. Pestivirus antibodies were found in three out of four large alpine ungulates in the High Valley of Susa. Seroprevalence was particularly high in chamois. Further investigation is needed to characterise the pestiviruses that circulate in these animals.     

Management of Wild Ungulate Populations in Italy: Captive-Breeding,Hybridisation and Genetic Consequences of Translocations

Captive-reproduced stocks of some species of ungulates (Artiodactyla), and particularly the red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus) and the wildboar (Sus scrofa) are more or less extensively translocated in Italy, mainly for local reintroductions or restocking of exploited wild populations. However, captive breeding often involves the reproduction of non-indigenous individuals or the production of artificial hybrids. Consequently, translocations of captive-reproduced ungulates are of concern for the conservation of indigenous populations and gene pools. The impact of translocations should be evaluated within the background of the growing knowledge on population genetic and phylogeographic structure of ungulates. Molecular genetic markers are being used to map geographic genetic diversity, and reconstruct the phylogeographic history of natural populations (i.e., in the roe deer). Molecular makers are also used to detect the consequences of domestication and identify hybrids between wild and domesticated populations (i.e., in the wildboar), or to detect inter-specific hybridisation (i.e., between the red deer and wapiti). Hybridisation of wild and domestic pigs, and diffusion of hybrids in nature is widespread in Italy. Admixture of indigenous and non-indigenous roe deer stocks is also widespread. Therefore, conservation and management of indigenous ungulates calls for careful evaluation of captive-reproduced stocks.

     

Serological surveillance for Tahyna virus (California encephalitis orthobunyavirus,Peribunyaviridae) neutralizing antibodies in wild ungulates in Austria,Hungary and Romania

A serosurvey for Tahyna virus (TAHV), a mosquito‐borne California encephalitis orthobunyavirus (Peribunyaviridae) endemic to Europe, was performed to estimate the activity of TAHV on a broad geographic scale. Sera from wild boar (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) were collected from Austria, Hungary and Romania. Samples were tested for neutralizing antibodies against TAHV using a virus microneutralization assay. The results demonstrate that TAHV transmission to mammals is widespread in Europe, particularly in the wild boar population where the mean rate of seroconversion is 15.2%.     

Faecal bacteria of wild ruminants and the alpine marmot

Faecal samples from 60 red deer (Cervus elaphus), 13 roe deer (Capreolus capreolus), 7 chamois (Rupicapra rupicapra), 41 alpine marmot (Marmota marmota) and soils mixed with deer faeces from the Stelvio National Park were examined forCampylobacter sp. andSalmonella sp. with negative results.The same material, especially deer faeces, was a habitat highly suitable forYersinia sp.:Y. enterocolitica (two biotypes) was isolated twice,Y. kristen-senii (two serotypes) was isolated 19 times,Y. frederiksenii andY. intermedia were isolated once.Antibiotic-resistantEscherichia coli were isolated from 16 specimens from wild ruminants, one from marmot and two from feeding places.This article was presented as a paper at the 8th International Symposium Enteric Infections and their Control of the World Association of Veterinary Microbiologists, Immunologists and Specialists in Infectious Diseases (Perth, Western Australia, 20 August 1983).     

Immunoglobulin G class identification from wild ungulates by cross-reactivity with antisera to domestic animals

Seven species of Spanish ungulates were tested for the presence of homologous immunoglobulin G (IgG) with a gel-diffusion test using bovine, ovine, caprine and porcine IgG antisera. Homologous ovine and caprine IgG were detected in sera from chamois (Rupicapra pyrenaica), Spanish ibex (Capra pyrenaica hispanica), mouflon (Ovis orientalis musimon), red deer (Cervus elaphus), fallow deer (Dama dama) and roe deer (Capreolus capreolus). Homologous porcine IgG was detected in wild boar (Sus scrofa) serum. Immunoelectrophoretic assays were performed to compare the electrophoretic mobility of IgG from domestic and wild species.     

Retrospective study of pestivirus infection in Pyrenean chamois (Rupicapra pyrenaica) and other ungulates in the Pyrenees (NE Spain)

In 2001 a new Pestivirus (Family Flaviviridae) was associated with an outbreak of a previously unreported disease in Pyrenean chamois (Rupicapra pyrenaica) in the Pyrenees (NE Spain). Molecular characterization assigned this virus to the Border Disease Virus (BDV) cluster, BDV-4 genotype. A retrospective study was performed in archived sera and spleen of 74 Pyrenean chamois and in archived sera of 28 mouflon (Ovis ammon), 56 red deer (Cervus elaphus), 43 roe deer (Capreolus capreolus) and 29 fallow deer (Dama dama) from the Pyrenees between the years 1990 and 2000. Thirty six of 74 (48.6%) sera of Pyrenean chamois, one of mouflon and one of red deer were positive by an ELISA antibody test. Comparative virus neutralization tests were performed on 26 seropositive chamois, one mouflon and one red deer, using five pestivirus strains. An ELISA antigen test was performed on 37 seronegative chamois and yielded positive results in one chamois and inconclusive result in two. RT-PCR and virus isolation performed on spleen samples from these three animals gave positive results in the positive and one inconclusive animal. Sequence analysis in the 5' unstranslated region revealed that they were grouped into the BDV-4 genotype. Virological and serological data of the present study indicate that BDV infection has been present in the chamois population since at least 1990, 11 years before the first outbreak of disease. Therefore, the emergence of the disease in 2001 is apparently due to other factors rather than the introduction of a new virus in the chamois population.     

Antimicrobial resistant bacteria in wild mammals and birds: a coincidence or cause for concern?

Background

The emergence and dissemination of antimicrobial resistance (AMR) is a growing concern to public and animal health. The contribution attributable to wildlife remains unclear. In this study two unrelated wildlife species herring gulls (Larus argentatus) and a hybrid deer (Cervus elaphus x Cervus nippon) were investigated for the presence of Escherichia coli expressing an AMR phenotype.

Findings

Bacterial isolates resistant to β-lactam compounds were identified in both animal species and the production of functional β-lactamase was confirmed using nitrocefin. The prevalence of resistant isolates was higher in herring gulls (87%) compared to deer (31%). Resistance to this class of antibiotic was found only in non-pathogenic E. coli in herring gulls and in both pathogenic and non-pathogenic E. coli strains in deer.

Conclusions

The presence of AMR in wildlife has implications for public health, food safety and potable water source protection among others.     

Gastrointestinal parasites in an isolated Norwegian population of wild red deer (Cervus elaphus)

Background

Thirteen red deer (Cervus elaphus), culled from the isolated population at the Mongstad Oil Refinery, Norway, were investigated for gastrointestinal helminths. These animals, enclosed by the refinery fence, do not have contact with other ruminants and have a high population density considering the available browsing area (1 km²) within the refinery site (3 km²). The population was estimated to be 110-130 at the time of culling.

Results

The helminth fauna among these sampled red deer was enumerated and species were identified based on morphology. Ostertagia leptospicularis/O. kolchida was detected in 83% [CI 55 - 95%], Spiculopteragia spiculoptera/S. mathevossiani in 92% [CI 65 - 99%] and Trichostrongylus axei in 42%, [CI 19 - 68%] of the abomasa examined. Characterisation of the intestinal parasite fauna revealed Capillaria bovis, Cooperia oncophora, Oesophagostomum venulosum, Trichuris globulosa and tapeworm fragments (presumed anoplocephalids) in seven individuals. Only one calf had an infection with more than one intestinal helminth (tapeworm fragment and Trichuris globulosa). The remaining six deer had single species intestinal infections.No significant age related trends were seen, with the exception of higher intensity of infection of T. axei in yearlings relative to other age classes. Assessment of abomasal parasite burden and body condition revealed no significant trends. In calves, statistically non-significant correlation was seen between increased parasite burden and decreased slaughter weight, whilst the opposite was seen in adults with the heaviest adults exhibiting the higher burdens. Given the small sample size the trends that were seen need further investigation. The parasite burden was aggregated with three adult red deer harbouring 75% of the total abomasal parasite count.

Conclusion

This isolated population was parasitised by a reduced subset of gastrointestinal nematodes typical of this cervid across an extensive geographic range in Eurasia. The intensity and abundance of abomasal nematodes was higher in this isolated population than reported in similar studies of red deer populations across Europe.     

Faecal bacteria of wild ruminants and the alpine marmot

Faecal samples from 60 red deer (Cervus elaphus), 13 roe deer (Capreolus capreolus), 7 chamois (Rupicapra rupicapra), 41 alpine marmot (Marmota marmota) and soils mixed with deer faeces from the Stelvio National Park were examined for Campylobacter sp. and Salmonella sp. with negative results. The same material, especially deer faeces, was a habitat highly suitable for Yersinia sp.: Y. enterocolitica (two biotypes) was isolated twice, Y. kristensenii (two serotypes) was isolated 19 times, Y. frederiksenii and Y. intermedia were isolated once. Antibiotic-resistant Escherichia coli were isolated from 16 specimens from wild ruminants, one from marmot and two from feeding places.     

Serologic and virologic investigations into pestivirus infection in wild and domestic ruminants in the Pyrenees (NE Spain)

An outbreak of disease associated to a border disease virus was described in the Southern chamois (Rupicapra pyrenaica) in Spain in 2002. Sera and/or spleen samples from 57 mouflon, 15 red deer, 21 roe deer, 3 fallow deer, 55 sheep, 32 cattle, and 68 goats sharing the chamois habitat were studied. An antibody ELISA test yielded an inconclusive result in 2 mouflon and positive results in 5 goat sera. Comparative virus neutralization tests were performed on the 2 inconclusive mouflons, 3 of the 5 seropositive goats, 55 sheep and 32 cattle, using 6 pestivirus strains. Positive results were obtained in 1 mouflon, 2 goats, 69% of sheep and 78% of cattle. Virological investigations performed with an antigen ELISA test yielded negative results in 21 goats and 39 mouflons, the result in 1 mouflon being inconclusive. PCR performed on 12 goats and the inconclusive mouflon gave negative results. These results suggested that it is unlikely that chamois BDV is infecting wild and domestic ruminants.     

Toxoplasma gondii and Neospora caninum in wildlife: common parasites in Belgian foxes and Cervidae?

Sera from Cervidae were tested for the presence of antibodies against Neospora caninum using ELISA; and against Toxoplasma gondii using SAG1-ELISA and a commercially available agglutination test. The T. gondii seroprevalence was 52% (38/73) in roe deer (Capreolus capreolus), 0% in bred fallow deer (0/4) (Dama dama) and red deer (0/7) (Cervus elaphus). We found 2.7% of the roe deer samples and none of the bred deer samples positive for N. caninum. Brain samples from wild roe deer, red deer and red foxes (Vulpes vulpes) were tested for the presence of T. gondii and N. caninum DNA using multiplex real-time PCR. We detected T. gondii in 18.8% (57/304) of the red foxes and in 1 of the 33 deer samples. N. caninum was found in 6.6% of the red foxes and in 2 roe deer samples. Twenty-six of the T. gondii positive DNA extracts from the red fox samples were genotyped. Twenty-five were type II and only one was found to be type III.     

Seroprevalence of Neospora caninum in non-carnivorous wildlife from Spain

Serum samples from 1034 non-carnivorous wildlife from Spain were tested for antibodies to Neospora caninum by competitive screening enzyme linked immunosorbent assay (ELISA) and confirmed by an indirect fluorescent antibody test (IFAT). High agreement was observed between results in both techniques (kappa value higher than 0.9). Prevalences of N. caninum antibodies positive by both techniques were 11.8% of 237 red deer (Cervus elaphus), 7.7% of 13 barbary sheep (Ammotragus lervia), 6.1% of 33 roe deer (Capreolus capreolus) and 0.3% of 298 wild boar (Sus scrofa). In one of 53 hares (Lepus granatensis), antibodies were found in the ELISA but could not be confirmed by IFAT due to lack of sample. Antibodies to N. caninum were not found in any of 251 wild rabbits (Oryctolagus cuniculus), 79 fallow deer (Dama dama), 27 mouflon (Ovis ammon), 40 chamois (Rupicapra pyrenaica) and three Spanish ibex (Capra pyrenaica). Statistically significant differences were observed between N. caninum seroprevalence in red deer and management of hunting estates (open versus fenced) with higher prevalence in fenced estates, and among sampling sites. Seroprevalence was particularly high in some areas (MO estate in South-Central Spain or some estates of Catalonia, North-East Spain), while no contact with N. caninum was observed in others. Results indicate that in certain areas of Spain, N. caninum is present in wildlife, especially in red deer. These results have important implications in both sylvatic cycles and may influence the prevalence of infection in cattle farms in those areas. To our knowledge, this is the first report of antibodies to N. caninum in wildlife from Spain and the first report of N. caninum antibodies in barbary sheep and wild boar.     

Sequence variations of the bovine prion protein gene (PRNP) in native Korean Hanwoo cattle

Bovine spongiform encephalopathy (BSE) is one of the fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs) caused by infectious prion proteins. Genetic variations correlated with susceptibility or resistance to TSE in humans and sheep have not been reported for bovine strains including those from Holstein, Jersey, and Japanese Black cattle. Here, we investigated bovine prion protein gene (PRNP) variations in Hanwoo cattle [Bos (B.) taurus coreanae], a native breed in Korea. We identified mutations and polymorphisms in the coding region of PRNP, determined their frequency, and evaluated their significance. We identified four synonymous polymorphisms and two non-synonymous mutations in PRNP, but found no novel polymorphisms. The sequence and number of octapeptide repeats were completely conserved, and the haplotype frequency of the coding region was similar to that of other B. taurus strains. When we examined the 23-bp and 12-bp insertion/deletion (indel) polymorphisms in the non-coding region of PRNP, Hanwoo cattle had a lower deletion allele and 23-bp del/12-bp del haplotype frequency than healthy and BSE-affected animals of other strains. Thus, Hanwoo are seemingly less susceptible to BSE than other strains due to the 23-bp and 12-bp indel polymorphisms.     

Prevalence of Toxoplasma gondii antibodies in wild red deer, roe deer, moose, and reindeer from Norway

Serum samples from 4339 wild cervids collected in Norway were tested for antibodies against Toxoplasma gondii using the direct agglutination test. The association between seroprevalence and species, sex, age, and geographic region was investigated. Positive titers (> or =1:40) were found in 33.9% of 760 roe deer (Capreolus capreolus); 12.6% of 2142 moose (Alces alces); 7.7% of 571 red deer (Cervus elaphus); and 1.0% of 866 reindeer (Rangifer tarandus). The seroprevalences were significantly different between the species. There was a significant increase in prevalence with age in roe deer, moose, and red deer, except from yearling to adult in red deer. A significant age-sex interaction was found in moose, and the effect of age was most distinct for females. No association between seropositive animals and sex was found for roe deer and red deer. There were significant differences in prevalence between geographic regions in roe deer and male moose. A widespread exposure to T. gondii in Norwegian cervids is documented, and meat from Norwegian cervids, particularly roe deer, should be regarded a potential source of infection for humans.     

Experimental in vitro transmission of Babesia sp. (EU1) by Ixodes ricinus

Babesia sp. (EU1), first characterized in 2003, has been implicated in human cases of babesiosis in Italy, Austria and Germany. It has been identified in roe deer and in its suspected tick vector, Ixodes ricinus, in several European countries. The aim of the present study was to validate the competence of I. ricinus as a vector of Babesia sp. (EU1) via experimental infections. For this purpose, a parasite strain isolated from roe deer was cloned in sheep erythrocytes. After experimental infections, parasite DNA was successfully amplified by PCR in both eggs and larvae originating from infected I. ricinus females and in the salivary glands of females exposed to Babesia sp. (EU1) as nymphs. We also demonstrate that infected females were able to transmit parasite DNA during a new blood meal. Together with previous epidemiological studies, these results validate I. ricinus as a competent vector for Babesia sp. (EU1).