Serologic survey for antibodies against Mycobacterium a vium subsp. paratuberculosis in free-ranging cervids from Norway

Affinity between protein-G and immunoglobulins from red deer (Cervus elaphus), moose (Alces alces), roe deer (Capreolus capreolus), and reindeer (Rangifer tarandus tarandus) was tested in a competition binding assay. Sera from red deer, reindeer, and moose inhibited the assay less than sera from cattle (less affinity), whereas sera from roe deer showed a slightly higher affinity to protein-G than did sera from cattle. The conclusion was made that protein-G could be used instead of anti-species antibodies for these cervid species, where the aim of the screening was to look for exposure or lack of exposure to mycobacteria in the tested populations. Serologic screening of 1,373 free-ranging cervids for antibodies against Mycobacterium avium subsp. paratuberculosis was conducted. All sera were tested by a protein-G-based antigen-absorbed enzyme-linked immunosorbent assay (ELISA). Seropositive moose (10/537; 1.9%), red deer (14/371; 3.8%), roe deer (6/49; 12.2%), and semidomesticated reindeer (11/325; 3.4%) were found, whereas wild reindeer (n = 91) were seronegative. In addition, the red deer sera were tested with a commercial ELISA, by which two animals tested positive and nine were suspicious of having M. avium subsp. paratuberculosis antibodies. Tissue samples and feces from 10 moose originating from a population with a clustering of seropositive animals were investigated by histology and bacteriology with negative results. Paratuberculosis has never been diagnosed in free-ranging or farmed cervid species in Norway. Thus, further studies are indicated to prove that the present findings reflect an infection with M. avium subsp. paratuberculosis.     

Serological survey of herpesvirus infections in wild ruminants of France and Belgium

The presence of antibodies against bovine herpesvirus 1 (BHV-1), bovid herpesvirus 6 (BHV-6), herpesvirus of Cervidae type 1 (HVC-1), reindeer herpesvirus, bovine herpesvirus 2 (BHV-2) and bovid herpesvirus 4 (BHV-4) was investigated in wild ruminants of France and Belgium between 1981 and 1986. There were no animals serologically positive for BHV-4. Antibodies against BHV-2 were demonstrated in roe deer (Cervus capreolus) (less than 1%) and chamois (Rupicapra rupicapra) (1%) in France. Animals seropositive to the four related viruses (BHV-1, BHV-6, HVC-1, reindeer herpesvirus) were detected in red deer (Cervus elaphus) in France and Belgium (1% and 11%, respectively), in roe deer (less than 1%) from France, in chamois (4%) in France and in ibex (Capra ibex) (4%) from France. The presence of antibodies against HVC-1, especially in red deer from Belgium, may suggest that wild ruminants in continental Europe are now infected with this virus, which previously has been isolated only in Scotland.     

Malignant catarrhal fever in free-ranging cervids associated with OvHV-2 and CpHV-2 DNA

Pathologic lesions were summarized in 18 free-ranging cervids (15 moose [Alces alces], two roe deer [Capreolus capreolus], and one red deer [Cervus elaphus]) diagnosed with malignant catarrhal fever (MCF) after examination at the National Veterinary Institute, Oslo 1982-2005. Eye lesions (conjunctivitis, corneal opacity, fibrin clots in the anterior eye chamber) were the most frequent gross finding. Erosive-ulcerative mucosal lesions in the nose and mouth were also commonly found. Histopathology revealed a nonpurulent vasculitis and perivasculitis in the central nervous system (CNS) typical of MCF in 16 of the cases. The diagnosis in the remaining two animals was based upon histologic eye lesions consistent with MCF (CNS not available for examination). Polymerase chain reaction was run on samples from 15 individuals for evidence of MCF-virus DNA, and ovine herpesvirus-2 (OvHV-2) DNA was detected in five moose, one roe deer, and one red deer, and caprine herpesvirus-2 (CpHV-2) DNA was detected in two moose and one roe deer. Sera from 1,000 free-ranging cervids were tested for specific antibodies to MCF-associated viruses (MCFV) by competitive inhibition enzyme-linked immunosorbent assay. The seroprevalences were: red deer 5%, reindeer (Rangifer tarandus) 4%, roe deer 2%, and moose 0.4% (n = 250 for all four species). The results indicate that sheep and goat MCFV may cause serious disease in wild moose, roe deer, and red deer. The seropositive cervids most likely represent individuals infected with either OvHV-2 or CpHV-2, but may also reflect infections with other related MCFV.     

Molecular identification and prevalence of Dictyocaulus spp. (Trichostrongyloidea: Dictyocaulidae) in Swedish semi-domestic and free-living cervids

Lungs of 102 roe deer (Capreolus capreolus), 136 moose (Alces alces), 68 fallow deer (Dama dama), and six red deer (Cervus elaphus) were examined during hunting seasons from 16 September 1997 to 1 March 2000. The aim was to determine the species composition and prevalence of Dictyocaulus lungworms in these hosts in Sweden. Worms were identified following polymerase chain reaction (PCR) amplification of the internal transcribed spacer of ribosomal DNA (ITS2), followed by hybridization with four species-specific oligonucleotides. In addition, 50 lungworms from five reindeer (Rangifer tarandus) from Norway were similarly analyzed. A total of 399 worms were recovered and analyzed representing a range of 29-128 worms per host species. All specimens from roe deer were identified as Dictyocaulus capreolus, whereas those from red deer and reindeer were identical with D. eckerti. From moose, 73 (81.1%) of the worms were identified as D. capreolus whereas 17 (18.9%) were D. eckerti. The ITS2 sequence of fallow deer lungworms differed significantly when compared with the ITS2 of D. viviparus, D. capreolus, and D. eckerti. This indicated that fallow deer in Sweden may be infected with a new genotype of Dictyocaulus spp. Consequently, a specific probe designed for the ITS2 from this Dictyocaulus sp. hybridized exclusively with samples from lungworms of fallow deer. Interestingly, no D. viviparus were found in any of these hosts. The prevalence of infection in each host was as follows: D. capreolus in roe deer (14.7%) and moose (10.6%); D. eckerti in moose (0.7%) and red deer (33.3%); and Dictyocaulus sp. in fallow deer (10.3%). Regardless of lungworm species, the overall prevalence of Dictyocaulus spp. in these hosts was 12.2%. Prevalence between male and female animals and among the different age groups did not differ significantly. Finally an enzyme linked immunosorbent assay (ELISA) specific for patent D. viviparus infection in cattle was utilized to analyze lung tissue fluids from infected animals. All samples from roe deer, red deer, and fallow deer were negative in the ELISA. However, three out of twelve (25%) samples from moose and 17 of 40 (43%) samples from cattle were positive. This indicated that moose anti-D. capreolus antibodies recognized the D. viviparus antigen and that anti-cattle immunoglobulin cross-reacted with moose antibodies.     

A comparative study of hepatic trace element levels in wild moose, roe deer, and reindeer from Norway

Liver samples from 422 wild moose (Alces alces), 280 roe deer (Capreolus capreolus), and 73 reindeer (Rangifer tarandus) collected by hunters in various localities in Norway, 2002-2003, were analyzed for the essential trace elements cobalt, copper (Cu), manganese (Mn), molybdenum, selenium (Se), and zinc. Significant differences in hepatic concentrations among species were found for all elements except for Mn, and considerable individual and geographic variations were seen. Roe deer had statistically significant lower Se levels (median: 0.51 μg Se/g dry weight) than did moose (0.77 μg Se/g) and reindeer (0.85 μg Se/g). Moose from two coastal municipalities with high precipitation had considerably higher Se concentrations than those from the other localities studied. Seventy-nine roe deer (28%) and 36 moose (9%) had Se concentrations below that regarded as deficient in domestic ruminants. The Se status in roe deer was lower than previously reported in Europe. Moose had a significantly higher Cu (222 μg Cu/g dw) than did roe deer (112 μg Cu/g) and reindeer (105 μg Cu/g). The Cu status of moose and roe deer in Norway are among the highest reported in Europe. However, a suboptimal Se and Cu status was found in moose from Tvedestrand, a population which has suffered from a reduced condition and productivity. The variability in trace element status among hunted cervids, with no apparent signs of deficiency or toxicity, probably reflects adaptations in these wild ruminant species to cope with this. However, subtle clinical signs and lesions are difficult to detect and further research is needed.     

Serological survey for potential disease agents of free-ranging cervids in six selected national parks from Germany

A total of 164 blood samples, collected from free-ranging red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in six German national parks (NP) between 2000 and 2002, were assayed for antibodies against nine viral disease agents. Antibodies were only detected against the alpha-herpesviruses; specifically, bovine herpesvirus-1 (BHV-1) (22 of 157, 14%), cervid herpesvirus-1 (17 of 157, 10.8%), and caprine herpesvirus-1 (11 of 159, 6.9%). Titers ranged from 4 to 102. Most of the seropositive sera, and those with the highest antibody titers, were from red and roe deer in the Harz and Hochharz NP, which are connected and allow migration between the two. The distribution and specificity of antibodies detected in individual deer suggests that the three alpha-herpesviruses are circulating in these deer populations. No antibodies were detected against bovine viral diarrhea virus, epizootic hemorrhagic disease virus, bovine leukemia virus, bluetongue virus, foot-and-mouth disease virus, or sheep and goat poxvirus.     

Experimental Narthecium ossifragum nephrotoxicity in cervids from Norway

One moose (Alces alces), two red deer (Cervus elaphus), two reindeer (Rangifer tarandus) and two fallow deer (Dama dama) were dosed intraruminally with an aqueous extract made from 30 g of bog asphodel (Narthecium ossifragum) (wet weight) per kg live weight. The moose and one of the two reindeer were mildly depressed and had reduced appetite 3 to 7 days and 1 to 4 days after dosing, respectively. The serum creatinine and urea concentrations increased markedly in the moose and red deer, and moderately in the reindeer. No increase in serum creatinine and urea was observed in the fallow deer. Histopathological examination of the kidneys of the animals, killed 8 to 10 days after dosing, revealed tubular epithelial cell degeneration, necrosis, and regeneration in the moose, red deer and reindeer. The renal lesions were severe in the moose, moderate in the red deer and mild in the reindeer. No histopathological lesions were seen in the kidneys of the fallow deer.     

The melanocyte-stimulating hormone receptor (MC1-R) gene as a tool in evolutionary studies of artiodactyles

The complete coding region of the melanocyte-stimulating hormone receptor (MC1-R) gene was characterized in species belonging to the two families Bovidae and Cervidae; cattle (Bos taurus), sheep (Ovis aries), goat (Capra hircus), muskox (Ovibos moschatus), roe deer (Capreolus capreolus), reindeer (Rangifer tarandus), moose (Alces alces), red deer (Cervus elaphus) and fallow deer (Dama dama). This well conserved gene is a central regulator of mammalian coat colour. Examination of the interspecies variability revealed a 5.3-6.8% divergence between the Cervidae and Bovidae families, whereas the divergence within the families were 1.0-3.1% and 1.2-4.6%, respectively. Complete identity was found when two subspecies of reindeer, Eurasian tundra reindeer (R.t. tarandus) and Svalbard reindeer (R.t. platvrhynehus), were analyzed. An rooted phylogenetic tree based on Bovidae and Cervidae MC1-R DNA sequences was in complete agreement with current taxonomy, and was supported by bootstrapping analysis. Due to different frequencies of silent vs. replacement mutations, the amino acid based phylogenetic tree contains several dissimilarities when compared to the DNA based phylogenetic tree.     

<Emphasis Type="Italic">Campylobacter</Emphasis> spp., <Emphasis Type="Italic">Salmonella</Emphasis> spp., Verocytotoxic <Emphasis Type="Italic">Escherichia coli</Emphasis>, and Antibiotic Resistance in Indicator Organisms in Wild Cervids

Faecal samples were collected, as part of the National Health Surveillance Program for Cervids (HOP) in Norway, from wild red deer, roe deer, moose and reindeer during ordinary hunting seasons from 2001 to 2003. Samples from a total of 618 animals were examined for verocytotoxic E. coli (VTEC); 611 animals for Salmonella and 324 animals for Campylobacter. A total of 50 samples were cultivated from each cervid species in order to isolate the indicator bacterial species E. coli and Enterococcus faecalis/E. faecium for antibiotic resistance pattern studies. Salmonella and the potentially human pathogenic verocytotoxic E. coli were not isolated, while Campylobacter jejuni jejuni was found in one roe deer sample only. Antibiotic resistance was found in 13 (7.3%) of the 179 E. coli isolates tested, eight of these being resistant against one type of antibiotic only. The proportion of resistant E. coli isolates was higher in wild reindeer (24%) than in the other cervids (2.2%). E. faecalis or E. faecium were isolated from 19 of the samples, none of these being reindeer. All the strains isolated were resistant against one (84%) or more (16%) antibiotics. A total of 14 E. faecalis -strains were resistant to virginiamycin only. The results indicate that the cervid species studied do not constitute an important infectious reservoir for either the human pathogens or the antibiotic resistant microorganisms included in the study.     

Febrile response and decrease in circulating lymphocytes following acute infection of white-tailed deer fawns with either a BVDV1 or a BVDV2 strain

Although commonly associated with infection in cattle, bovine viral diarrhea viruses (BVDV) also replicate in many domestic and wildlife species, including cervids. Bovine viral diarrhea viruses have been isolated from a number of cervids, including mule deer (Odocoileus hemionus), European roe deer (Capreolus capreolus), red deer (Cervus elaphus), white-tailed deer (Odocoileus virginianus), and mouse deer (Tragulus javanicus), but little information is available regarding clinical presentation and progression of infection in these species. In preliminary studies of experimental infection of deer with BVDV, researchers noted seroconversion but no clinical signs. In this study, we infected white-tailed deer fawns that were negative for BVDV and for antibodies against BVDV, with either a type 1 or a type 2 BVDV that had been isolated from white-tailed deer. Fawns were monitored for changes in basal temperature, circulating lymphocytes, and platelets. The clinical progression following inoculation in these fawns was similar to that seen with BVDV infections in cattle and included fever and depletion of circulating lymphocytes. Because free-ranging cervid populations are frequently in contact with domestic cattle in the United States, possible transfer of BVDV between cattle and cervids has significant implications for proposed BVDV control programs.     

Antibodies to granulocytic Ehrlichia in moose, red deer, and roe deer in Norway

Serum samples from 104 moose (Alces alces), 124 red deer (Cervus elaphus) and 114 roe deer (Capreolus capreolus), collected from different counties in southern Norway from 1994 to 2000, were analysed by an indirect immunofluorescent antibody staining method for antibodies to Ehrlichia equi. The overall seroprevalences for granulocytic Ehrlichia spp. in moose, red deer, and roe deer from Ixodes ricinus infested counties were 43%, 55%, and 96%, respectively. Antibody prevalence was significantly higher in roe deer than in moose and red deer (P < 0.001). Mean antibody titers (log10 +/- SD) to E. equi in sera from moose, red deer, and roe deer were 1:1,497 (3.17 +/- 0.646), 1:234 (2.37 +/- 0.424) and 1:676 (2.83 +/- 0.404), respectively. The present work indicates that all these wild ruminant species are exposed to granulocytic Ehrlichia in Norway.     

Ultrastructure of the cysts of Sarcocystis grueneri from cardiac muscle of reindeer (Rangifer tarandus tarandus)

Cysts of Sarcocystis grueneri from cardiac muscle of reindeer (Rangifer tarandus) in Norway were examined by transmission electron microscopy. The limiting unit membrane of the cyst proper formed regularly spaced invaginations into the cyst at numerous sites coinciding with interruptions in the underlying osmiophilic layer. The primary cyst wall formed numerous strip-like, sinuous protrusions, which were 30-40 nm thick, 150-300 nm wide and up to 4.5 microns long, and were running in parallel with the surface of the cyst. Generally the protrusions were arranged in several closely spaced layers compressed against the cyst. The nature and arrangement of the protrusions render them undetectable by light microscopy. Cyst ground substance divided the interior of the cyst into compartments containing typical sarcosporidian metrocytes and cystozoites. The cysts of S. grueneri from reindeer were ultrastructurally similar to cysts reported from red deer, roe deer and moose by other workers. The possibility that these cervids are hosts for a common Sarcocystis species is discussed.     

Prevalence of Neospora caninum and Toxoplasma gondii antibodies in wild ruminants from the countryside or captivity in the Czech Republic

In the Czech Republic, sera from 720 wild ruminants were examined for antibodies to Neospora caninum by screening competitive-inhibition enzyme-linked immunosorbent assay and confirmed by indirect fluorescence antibody test (IFAT); the same sera were also examined for antibodies to Toxoplasma gondii by IFAT. Neospora caninum antibodies were found in 14% (11 positive/79 tested) roe deer (Capreolus capreolus), 14% (2/14) sika deer (Cervus nippon), 6% (24/ 377) red deer (Cervus elaphus), 1% (2/143) fallow deer (Dama dama), 3% (3/105) mouflon (Ovis musimon), and none of 2 reindeer (Rangifer tarandus). Toxoplasma gondii antibodies were found in 50% (7/14) sika deer, 45% (169/377) red deer, 24% (19/79) roe deer, 17% (24/143) fallow deer, 9% (9/105) mouflon, and 1 of 2 reindeer. In 42 samples of wild ruminants that tested positive for N. caninum antibodies, 28 (67% of the positive N. caninum samples) reacted solely to N. caninum. This is the first evidence of N. caninum infection in mouflon, the first N. caninum seroprevalence study in farmed red deer, and the first survey of N. caninum in wild ruminants from the Czech Republic.     

黑龙江省胜山林场冬季驼鹿、马鹿和狍的种间关系

1987—1989年的两个冬季在黑龙江省黑河市胜山林场,对驼鹿、马鹿和狍的食性、采食高度和栖息地进行了研究。驼鹿的主要食物是柳(22.4％)、榛(16.0％)、红松(14.3％)和桦(13.8％);马鹿为杨(36.0％)、桦(22.7％)和红松(17.3％);狍为桦(26.5％)、柳(19.0％)、杨(12.7％)。三种动物采食高度的频次为正态分布。驼鹿的采食高度(标准差)为137.4(±39.0)厘米;马鹿为136.2(±37.0)厘米;狍为84.2(±29.2)厘米。驼鹿和马鹿对红松林的利用程度最高,而狍主要利用沼泽植物和红松林。驼鹿与狍在食物维和栖息地维上的生态位重叠最大,驼鹿与马鹿在采食高度维上的重叠最大。三种动物在食物、采食高度和栖息地三维生态位上的重叠大小的顺序为:驼鹿和马鹿>驼鹿和狍>马鹿和狍。     

Serosurvey of roe deer, chamois and domestic sheep in the central Italian Alps

Roe deer (Capreolus capreolus), chamois (Rupicapra rupricapra rupicapra), and domestic sheep in the Orobie Alps, Italy, were serologically tested for antibodies to selected pathogens that may be transmitted across species. Antibodies against Brucella spp. and bovine herpesvirus 1 (roe deer and chamois only) were not detected in any species. In roe deer, antibodies were detected against Toxoplasma gondii (13%) and Neospora caninum (3%). Chamois tested positive for antibodies to T. gondii (5%), N. caninum (21%), bovine respiratory syncytial virus (BRSV) (41%), bovine parainfluenza type-3 virus (17%), pestiviruses (18%), and Mycoplasma conjunctivae (17%). In the sheep, particularly high antibody prevalence rates were found for T. gondii (78%), Chlamydophila spp. (20%), pestiviruses (90%), BRSV (82%), and M. conjunctivae (81%).     

PCR detection of bovine herpesviruses from nonbovine ruminants in Hungary

Polymerase chain reaction (PCR) was used to test six different nonbovine ruminant species for five bovine herpesviruses including infectious bovine rhinotracheitis virus (BoHV-1), bovine herpes mammillitis virus (BoHV-2), Movar-type herpesvirus (BoHV-4), bovine herpesvirus type 5 (BoHV-5), and alcelaphine herpesvirus 1 (AlHV-1). Species tested included 56 roe deer (Capreolus capreolus), 66 red deer (Cervus elaphus), 20 fallow deer (Dama dama), 16 mouflon (Ovis musimon), 34 domestic sheep, and 44 domestic goats, which were sampled in Hungary in 2003. Tracheal and popliteal lymph nodes collected from these animals were tested for the presence of the five bovine herpesviruses using three nested (two of which were duplex) PCR assays. Three bovine herpesviruses (BoHV-1, -2, and -4) were detected, whereas no evidence of AlHV-1 or BoHV-5 was observed. Prevalence of BoHV-1 ranged from 12% to 47%, and PCR-positive results were observed in all species tested. BoHV-2 was detected from roe deer, red deer, fallow deer, mouflon, and domestic sheep, and the prevalence in these species ranged from 3% to 50%. BoHV-4 was detected in all species, with prevalence ranging from 12% to 69%. Sequenced PCR products were 99-100% identical to bovine herpesviral sequences deposited in the GenBank.     

Polymorphisms and variants in the prion protein sequence of European moose (Alces alces), reindeer (Rangifer tarandus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in Scandinavia

The prion protein (PrP) sequence of European moose, reindeer, roe deer and fallow deer in Scandinavia has high homology to the PrP sequence of North American cervids. Variants in the European moose PrP sequence were found at amino acid position 109 as K or Q. The 109Q variant is unique in the PrP sequence of vertebrates. During the 1980s a wasting syndrome in Swedish moose, Moose Wasting Syndrome (MWS), was described. SNP analysis demonstrated a difference in the observed genotype proportions of the heterozygous Q/K and homozygous Q/Q variants in the MWS animals compared with the healthy animals. In MWS moose the allele frequencies for 109K and 109Q were 0.73 and 0.27, respectively, and for healthy animals 0.69 and 0.31. Both alleles were seen as heterozygotes and homozygotes. In reindeer, PrP sequence variation was demonstrated at codon 176 as D or N and codon 225 as S or Y. The PrP sequences in roe deer and fallow deer were identical with published GenBank sequences.     

Polymorphisms and variants in the prion protein sequence of European moose (Alces alces), reindeer (Rangifer tarandus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in Scandinavia

The prion protein (PrP) sequence of European moose, reindeer, roe deer and fallow deer in Scandinavia has high homology to the PrP sequence of North American cervids. Variants in the European moose PrP sequence were found at amino acid position 109 as K or Q. The 109Q variant is unique in the PrP sequence of vertebrates. During the 1980s a wasting syndrome in Swedish moose, Moose Wasting Syndrome (MWS), was described. SNP analysis demonstrated a difference in the observed genotype proportions of the heterozygous Q/K and homozygous Q/Q variants in the MWS animals compared with the healthy animals. In MWS moose the allele frequencies for 109K and 109Q were 0.73 and 0.27, respectively, and for healthy animals 0.69 and 0.31. Both alleles were seen as heterozygotes and homozygotes. In reindeer, PrP sequence variation was demonstrated at codon 176 as D or N and codon 225 as S or Y. The PrP sequences in roe deer and fallow deer were identical with published GenBank sequences.