Prevalence of antibodies against Toxoplasma gondii and Neospora caninum in moose (Alces alces) and roe deer (Capreolus capreolus) in Sweden

Toxoplasma gondii and Neospora caninum are two coccidian parasites with a worldwide distribution. T. gondii is one of the more common parasitic zoonoses in the world and in young children and immunocompromised persons, infection can lead to severe disease and death. N. caninum is an important cause of abortions in cattle. Wildlife have been identified as reservoirs and transmitters for both parasites. The purpose of this study was to investigate the seroprevalences of T. gondii, and N. caninum in moose (Alces alces), and roe deer (Capreolus capreolus) in Sweden. Blood samples were collected from 417 moose during 2000-2005 and from 199 roe deer during 1990-2007. The samples were investigated for presence of antibodies by a T. gondii direct agglutination test and a N. caninum iscom ELISA. Because the iscom ELISA has not been validated for moose or roe deer, sera that gave a positive result were further investigated by immunoblot analysis to verify presence of antibodies. Antibodies to T. gondii were detected in 85 (20%) and 68 (34%) moose and roe deer sera, respectively. In moose the seroprevalence was higher in south and central Sweden than in the north, whereas there was no difference between the regions for roe deer. Adult moose and roe deer had higher odds of being seropositive than young animals but there were no difference in seroprevalence between males and females. One roe deer was positive by immunoblotting and was regarded as N. caninum positive, whereas all moose sera were negative. The results show that T. gondii infection is widely spread in the Swedish moose and roe deer populations. Precautions should therefore be taken when handling internal organs and carcasses of harvested cervids. Proper handling and cooking of game meat also is important to prevent toxoplasmosis in humans.     

Prevalence of Coxiella burnetti infection in wild and farmed ungulates

The aim of this study was to evaluate by serology and PCR analyses the prevalence of Coxiella burnetti infection in ungulates in Spain. Sera were collected from red deer (Cervus elaphus; n=116), roe deer (Capreolus capreolus; n=39), fallow deer (Dama dama; n=13) and cattle (n=79). Sera were tested for anti-C. burnetii antibody detection by means of an immunofluorescence antibody assay (IFA) and C. burnetii DNA was amplified by PCR in samples from ungulates that had antibodies to phase II antigens. Twenty-nine, 15 and 39 percent of the red deer, roe deer and cattle had antibodies against C. burnetii, respectively. None of the fallow deer sera tested positive. Seroprevalence was statistically higher in farmed than in wild red deer and higher in northern than in southern populations, whereas an inverse pattern was observed for the roe deer. Most of the seropositive animals had only anti-C. burnetii phase II antibodies, thus showing the acute nature of infections in the sampled ungulates. These results show that C. burnetii circulates in wild ungulates in Spain and suggest that they can act as pathogen reservoirs for both domestic animals and humans.     

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.     

Evidence of Anaplasma infections in European roe deer (Capreolus capreolus) from southern Spain

Anaplasma spp. (Rickettsiales: Anaplasmataceae) are tick-borne pathogens of veterinary and human importance. The wildlife hosts for these pathogens are not well characterized and may play an important role in the epidemiology of the disease. The objective of this research was to study the infection with A. marginale, A. ovis and A. phagocytophilum in free-ranging European roe deer (Capreolus capreolus) from Cádiz, Andalucía, Spain. Of 17 roe deer tested, 14 (82%) and 5 (29%) had antibodies reactive to Anaplasma spp. and A. phagocytophilum by competitive ELISA and indirect immunofluorescent antibody testing, respectively. Polymerase chain reaction and sequence analysis of Anaplasma major surface protein 4 (msp4) gene was conducted on blood samples from all roe deer examined. Nine (53%) animals had evidence of infection with A. ovis and 3 (18%) were positive for A. phagocytophilum. Concurrent infections were not detected. Despite the presence of A. marginale infections in cattle in the study site (36% msp4 PCR-positive animals), none of the msp4 amplicons from roe deer corresponded to A. marginale sequences. A. ovis msp4 sequences were identical to a genotype previously identified in sheep in Sicily, Italy. Two different A. phagocytophilum genotypes were identified in infected roe deer. This is the first report of roe deer naturally infected with A. ovis. These results demonstrate that roe deer are infected with A. ovis and A. phagocytophilum in Spain and suggest that this species may be involved in the natural cycle of these pathogens in this region, thus acting as potential reservoir for transmission to domestic and wild animals.     

Serum biochemical values of rusa deer (Cervus timorensis russa) in New Caledonia

Blood samples were collected from 91 rusa deer (Cervus timorensis russa), immediately after being shot. Serum mean biochemical values from shot deer are presented for blood urea nitrogen, creatinine, creatine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, total protein, albumin, calcium, and phosphorus. Mean total protein and albumin increased with age. There was an age-associated increase of gamma globulins. Mean creatine kinase activity and creatinine, albumin and phosphorus concentrations were higher in stags than in hinds. Pregnant hinds had lower mean creatine kinase activity and phosphorus and higher mean alanine aminotransferase and total protein than non-pregnant hinds. Mean calcium concentration increased when deer were agitated before bleeding.     

Prevalence of Cryptosporidium and Giardia in roe deer (Capreolus capreolus) and wild boars (Sus scrofa) in Galicia (NW, Spain)

Faecal samples from 224 roe deer (Capreolus capreolus) and 381 wild boars (Sus scrofa) shot during the 2008-2009 hunting season (August-January) in Galicia (NW Spain) were examined to determine the presence and intensity of infection by Cryptosporidium and Giardia. Analysis of a single sample from each of the roe deer revealed that the prevalence of cryptosporidiosis and giardiosis was 1.3% and 5.3% respectively. The prevalence of Giardia infection was significantly higher in juvenile female roe deer than in adult females, but no other significant differences were found in relation to age and sex. In wild boars, the prevalence of cryptosporidiosis and giardiosis was 7.6% and 1.3% respectively. The prevalence of Cryptosporidium infection was significantly higher in juvenile male wild boars than in adult males, but no other significant differences were found in relation to age or sex. In both groups of wild animals, the number of Cryptosporidium oocysts per gram of faeces (OPG) ranged from 5 to 200 and the number of Giardia cysts per gram of faeces (CPG) was between 5 and 47; there were no significant differences between the two groups with respect to number of infections. This is the first large study of Cryptosporidium and Giardia in roe deer and wild boars in hunting areas in Spain and the results demonstrate a low, but widespread prevalence of Cryptosporidium and Giardia in these animals.     

Effect of acepromazine on the signs of capture stress in captive and free-ranging roe deer (Capreolus capreolus)

The differences between the capture stress responses of captive and free-ranging roe deer (Capreolus capreolus) and their modulation by acepromazine, during a period of three hours' physical restraint after capture in drive-nets, were examined in 16 free-ranging and 16 captive roe deer. Eight of the free-ranging and eight of the captive animals received acepromazine intramuscularly, and the other eight free-ranging and eight captive deer received the same volume of saline. Heart rate, body temperature and haematological and serum biochemical parameters were analysed. In the groups treated with acepromazine, the heart rate stabilised sooner, and the red blood cell (RBC) count, haemoglobin concentration, packed-cell volume, the serum activities of creatine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) and the concentrations of creatinine and lactate were significantly lower, and serum glucose started to decrease earlier, than in the untreated groups. Serum potassium levels decreased over time only in the untreated groups. The body temperature stabilised earlier, and the RBC count, haemoglobin concentration, serum CK, AST, ALT and LDH activities, and serum creatinine, lactate, cholesterol and glucose concentrations were significantly lower in the free-ranging roe deer than in the captive deer.     

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.     

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.     

Prevalence of antibodies against Toxoplasma gondii in roe deer from Spain

Roe deer (Capreolus capreolus) is an important game animal in Spain. Sera from 278 roe deer from eight areas in mainland Spain were assayed for antibodies to Toxoplasma gondii by modified agglutination test (MAT). Titers of 1:25 or higher were found in 109 (39.2%) of 278 deer. No significant differences in antibody prevalence were found between sex or age categories. In contrast, significant differences in seroprevalence between locations were evident. Roe deer from the Northern coastal habitats (high humidity and roe deer density) had the highest prevalence, compared with low prevalence in Central Spain (arid areas and low roe deer density). There was a positive correlation between antibody prevalence and mean annual rainfall (r(s)=0.85, n=8, P<0.01). These findings have environmental and/or public health implications because venison can be an important meat source of T. gondii infections for humans and feral cats.     

Prevalence of Cryptosporidium and Giardia in free-ranging wild cervids in Norway

Faecal samples were collected from 1,190 wild cervids in Norway and analyzed for cysts/oocysts of the protozoan parasites Giardia and Cryptosporidium. Samples were from calves, yearlings and adults of moose (Alces alces), red deer (Cervus elaphus), roe deer (Capreolus capreolus) and reindeer (Rangifer tarandus) shot during the hunting season. Cryptosporidium was found in 15 (3.3%) of 455 moose, 1 (0.3%) of 289 red deer, 18 (6.2%) of 291 roe deer, but was not found in any of 155 reindeer. Giardia was found in 56 (12.3%) moose, 5 (1.7%) red deer, 45 (15.5%) roe deer and 11 (7.1%) reindeer. The calves had the highest prevalence of infection, but this was only statistically significant for Giardia in moose and for Cryptosporidium and Giardia in roe deer. Calves generally had the highest intensity of infection, but this difference was only statistically significant for calves with Giarda and the highest intensity of infection. Both Giardia and Cryptosporidium were found in samples from several geographical areas, indicating that these parasites are distributed among the cervid population in all parts of Norway, especially in moose and roe deer. This is the first published report of Cryptosporidium in moose and of Giardia in reindeer.     

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.     

不同季节日粮组成对狍瘤胃内环境的影响

选用8只健康成年狍,采用瘤胃穿刺法收集瘤胃液。研究不同季节日粮组成对狍瘤胃pH、挥发性脂肪酸（VFAs）、氨态氮（NH3-N）、茵体蛋白（BCP）、内毒素以及组织胺浓度等瘤胃内环境指标的影响。结果显示：夏季日粮与冬季日粮相比较,瘤胃中NH3-N和BCP浓度显著升高（P〈0．05）;瘤胃pH和乙酸水平显著降低（P〈0．05）;丙酸水平显著提升（P〈0．05）;而内毒素和组织胺浓度无显著变化（P〉0．05）。试验结果表明,冬、夏日粮组成均能够满足狍的营养需要以及稳定狍的瘤胃内环境,但夏季日粮组成对狍瘤胃发酵和营养需要更为适宜。     

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.

     

人工养殖西伯利亚狍肠道微生物组成的雌雄差异

肠道微生物是一个复杂的生态系统,与宿主的健康、疾病以及其他因素有着密切的关系。目前,关于肠道菌群结构和功能的研究较多,但是对西伯利亚狍肠道菌群的研究还未见报道。西伯利亚狍(Capreolus pygargus)属于反刍动物,有独特的消化特性和微生物群来帮助它们适应高纤维的食物。本研究利用高通量测序技术在Illumina MiSeq平台上对14只狍肠道微生物进行16S rRNA基因分析。结果表明:个体样本中门类组成的差异,雌性个体JY3和雄性个体JY9除Firmicutes,Bacteroidetes,Proteobacteria这3类菌群外,其他门类菌群所占比例都很低。在科水平上,雌性个体中Ruminococcaceae在JY7和JY13肠道内含量相对较少,Bacteroidaceae在JY7,JY10和JY13中含量较高。雄性个体中Ruminococcaceae在JY11和JY14肠道内含量较少,Prevotellaceae在JY4中含量较少。在属水平上,雌性个体特有的菌属为Roseburia,Alloprevotella,Lachnospiraceae_incertae_sedis和Coprococcus;雄性个体特有的菌属为Fusobacterium,Succiniclasticum,Streptococcus和Clostridium sensu stricto。Alpha多样性分析表明,Ace指数范围2128.93—772.98,Chao1指数范围2128.89—785.27,Shannon指数范围5.44—3.78,Simpson指数范围0.07—0.01。在不同物种分类水平上,雌雄狍肠道内的优势菌群有些不同。PCoA(principal co-ordinates analysis)分析显示,狍样本几乎混在一起,没有明显差异。     

The incidence of bacterial CNS infections in roe deer (Capreolus capreolus), red deer (Cervus elaphus) and chamois (Rupicapra rupicapra) in Bavaria

Brain samples of 849 wild ruminants (654 roe deer, 189 red deer and 6 chamois) from Bavaria were examined for the occurrence of encephalopathies caused by bacteria, using cultural, serological and genetic methods. In addition, 87 brain samples were investigated histologically for clarification of the pathogenetic relevance of specific microorganisms. Using conventional bacteriological methods, 464 different bacteria were isolated. 229 of them could be differentiated to the genus level and 235 to the species level. Totally, 35 different bacteria species were isolated, most frequently Micrococcus spp., Bacillus spp. and E. coli. Listeria spp. were detected in 43 brain samples (37 from roe deer, 5 from red deer and 1 from chamois). Sixteen strains were identified as L. innocua, 14 as L. monocytogenes, 9 as L. seeligeri and 4 as L. grayi. Serological investigations of L. monocytogenes showed that 9 strains belong to serotype 1/2a and five to 4b. Analysis of the geographical distribution of the Listeria findings indicate a statistically significant (p<0.011) regional aggregation in Unterfranken (prevalence for roe deer: 12.2%, versus 4.5% in Oberbayern-Schwaben, 6.1% in Niederbayern-Oberpfalz and 0% in Oberfranken-Mittelfranken). The histological investigation (HE staining) of 87 tissue samples contaminated with encephalitis relevant bacteria showed inflammation of different severity (mild meningitis and choroiditis (n = 26) to moderate (meningo)encephalitis (n = 13)) in 41 cases.     

Serum proteins of normal goats and goats with caseous lymphadenitis.

Values for total serum proteins and relative percentages of albumin, alpha 1-globulin, alpha 2-globulin, beta-globulin, and gamma-globulin were determined for the goat. These normal values were compared with those obtained for goats infected with Corynebacterium pseudotuberculosis. Goats chronically infected with C pseudotuberculosis show significantly higher total serum protein values than normal goats, apparently due to increased gamma-globulins. This higher protein value is also associated with a decrease in serum alpha 2- and beta-globulins.     

The occurrence of transmissible spongiform encephalopathies in roe deer (Capreolus capreolus), red deer (Cervus elaphus) and chamois (Rupicapra rupicapra) in Bavaria

Brain samples of 849 free-ranging ruminants (654 roe deer, 189 red deer and 6 chamois) from Bavaria were examined for TSE. In this respect, detections of PrPres were carried out by ELISA technique (BioRad) and immunohistochemistry using the monoclonal antibody (mAb) L42. Validation experiments of the ELISA technique for the detection of prionprotein from free-ranging ruminants showed that the examination of brains of wild ruminants lead to lower extinctions than that of bovine brain samples. Due to the fact that brain samples of 2 known CWD positive wapitis showed definite positive results, all samples were analysed by this test. There were no indications on the occurrence of TSE in any of the sample. In addition, immunohistochemical investigations of 10% of all samples did not show PrPres. Due to the large number of samples, the prevalence of TSE in Bavaria can be quoted under 0.5% for roe deer and under 1.5% for red deer with a certainty of 95%.     

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.     

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.