Epidemiology of an outbreak of chronic wasting disease on elk farms in Saskatchewan

An outbreak of chronic wasting disease (CWD) in farmed elk in Saskatchewan from 1996 to 2002 was reviewed to 1, determine the progression of CWD from infection to death in farmed elk; 2, assess animal risk factors for CWD infection in farmed elk; 3, assess farm management and exposure risk factors for within herd CWD transmission; and 4, assess the suitability of the Canadian Food Inspection Agency's (CFIA) current disease control policy for CWD in light of the findings. The results from animal movement tracing, animal testing, and a farm management questionnaire were used. The duration of CWD (time from exposure to death of a CWD test-positive animal) was between a mean minimum of 19 months and a mean maximum of 40 months. Age and sex were not associated with CWD infection, except that adult elk (> or = 2 y) were more likely to be infected than young elk (< 18 mo) (RR = 2.3, 95% CI 1.6-3.5). Elk calves born in the last 18 mo prior to the death or diagnosis of their dam were at higher risk if their dams died of CWD (RR = 4.1, 95% CI 1.5-11.4) or exhibited clinical signs of CWD (RR = 8.3, 95% CI 2.7-25.7). Significant risk factors for transmission of CWD on elk farms were the introduction from an infected farm of trace-in elk that died of CWD (RR = 13.5, 95% CI 2.0-91) or developed clinical signs of CWD (RR = 7.1, 95% CI 0.93-54) and the elapsed time in years since the incursion of CWD (OR = 5.6, 95% CI 1.8-17.4). The assumptions on which CFIA's disease control policies were based were validated, but based on this new information, quarantine in cases where exposure to preclinical elk has occurred could be considered as an alternative to whole herd eradication.     

The first Canadian indigenous case of bovine spongiform encephalopathy (BSE) has molecular characteristics for prion protein that are similar to those of BSE in the United Kingdom but differ from those of chronic wasting disease in captive elk and deer

Brain tissue from a case of bovine spongiform encephalopathy (BSE) from Alberta was subjected to a Western immunoblotting technique to ascertain the molecular profile of any disease-specific, abnormal prion protein, that is, prion protein that is protease-resistant (PrP(res)). This technique can discriminate between isolates from BSE, ovine scrapie, and sheep experimentally infected with BSE. Isolates of brain tissue from the BSE case in Alberta, 3 farmed elk with chronic wasting disease (CWD) from different parts of Saskatchewan, and 1 farmed white-tailed deer with CWD from Edmonton, Alberta, were examined alongside isolates of brain tissue from BSE, ovine scrapie, and sheep experimentally infected with BSE from the United Kingdom (UK). The molecular weights of PrP(res) and the cross reactions to 2 specific monoclonal antibodies (mAbs) were determined for each sample. The BSE isolates from Canada and the UK had very similar PrP(res) molecular weights and reacted with only 1 of the 2 mAbs. The PrP(res) isolated from both elk and white-tailed deer with CWD had a higher molecular weight profile than did the corresponding PrP(res) from the scrapie and BSE isolates. The PrP(res) from CWD cases cross reacted with both mAbs, a property shared with PrP(res) in isolates from scrapie but not with PrP(res) isolates from BSE or sheep experimentally infected with BSE. The results from this study seem to confirm that the PrP(res) isolated from the BSE case in Alberta has similar molecular properties to the PrP(res) isolated from a BSE case in the UK, and that it differs in its molecular and immunological characteristics from the CWD and scrapie cases studied.     

Chronic wasting disease in deer and elk: scientific facts and findings

Chronic wasting disease (CWD) is a prion disease of cervids such as deer and elk in North America. Unlike other transmissible spongiform encephalopathy (TSE) such as scrapie, CWD occurs in both captive and wild ranging animals, but not in domestic ruminants such as sheep and cattle. In this paper, the history of the disease, pathogenesis of CWD, susceptibility of animals, its transmission mechanisms, potential origins of the disease, diagnostic methods in the field and laboratory tests, surveillance and survey systems in the USA and Canada, control strategies, economic impact of the disease, food and feed safety, and the risks in human and animals are reviewed and summarized. Although there is no evidence that CWD has been transmitted to humans, it may have the potential to infect humans.     

A case of chronic wasting disease in an elk imported to Korea from Canada

A seven-year-old male elk (Cervus elaphus nelsoni) was euthanized and necropsied after having a 3-week history of body weight loss, emaciation, excessive salivation, teeth grinding, fever, anorexia, and respiratory distress. The elk was imported into Korea from Canada on March 9, 1997. Gross pathologic findings were restricted to a diffuse fibrinous pneumonia. Microscopic lesions included mild neuronal vacuolation and spongiform change in the neuropil of selected brain stem nuclei and generalized astrocytosis. Immunohistochemistry for protease-resistant prion protein (PrP(res)) was positive in all brain sections but more pronounced in the section of the obex of the medulla. And the PrP(res) was also detected by western immunoblotting in the brain and spinal cord. All the remaining elk and deer that had been in contact with this elk were destroyed and negative for chronic wasting disease (CWD). To our knowledge, this is the first case of CWD occurring outside of the U.S.A. and Canada.     

Pathological characterization of TgElk mice injected with brain homogenate from elk with chronic wasting disease

Chronic wasting disease (CWD) is classified as a transmissible spongiform encephalopathy or prion disease that affects cervids. CWD has been reported in 15 US states, two Canadian provinces, and in imported elk on several farms in Korea. This study was conducted to examine the molecular biological and pathogenic characteristics of a CWD-associated prion isolated in Korea. The epidemiological origin of this pathogen was also determined. Homozygous TgElk mice were infected with a CWD-affected elk brain pool prepared from the brain of an imported Canadian elk. We measured the incubation time of the pathogen, neuropathological changes by immunohistochemical staining, the pattern(s) of scrapie prion protein (PrP^Sc) deposition, and PrP^Sc protein profiles by Western blotting. We found that TgElk mice infected with brain homogenate from the elk suffering from CWD showed incubation times, vacuolar degeneration, and PrP^Sc accumulation similar to those previously reported in the literature. Our results suggest that homozygous TgElk mice efficiently transmit CWD with short incubation times and that this animal can serve a valuable research model and reliable in vivo diagnostic tool.     

First chronic wasting disease (CWD) surveillance of roe deer (Capreolus capreolus) in the northern part of Belgium

Cases of chronic wasting disease (CWD) in wild cervids have yet not been reported in Europe, whereas the disease is considered enzootic in free-ranging mule deer, Rocky mountain elk and white-tailed deer in the area of Colorado, Wyoming, and Nebraska. New foci of CWD continue to be detected in other parts of the United States. However, no large-scale active epidemiosurveillance of European wild cervids is yet installed in Europe. In accordance with the opinion of the European Scientific Steering Committee, a preliminary (active) surveillance scheme was installed, in order to improve the knowledge of the CWD status of wild cervids (roe deer) in the Northern part of Belgium. Spleen samples (n=206) and brain samples (n=222) of roe deer collected in the Northern part of Belgium, were examined for CWD using the antigen-capture enzyme-linked immunoassay (EIA) of IDEXX. Afterwards, the EIA was systematically confirmed by immunohistochemistry using three antibodies, namely R524, 2G11 and 12F10. There were no indications on the occurrence of TSE in any of the samples. A Bayesian framework was used for the estimation of the true prevalence of CWD in the Northern part of Belgium that was estimated to have a median value of zero with a 95th percentile value of 0.0049 and 0.0045 for spleen and brain samples respectively.     

Immunohistochemical diagnosis of chronic wasting disease in preclinically affected elk from a captive herd.

An immunohistochemical (IHC) method was used to test brain tissues from 17 elk in a captive herd in which chronic wasting disease (CWD) had previously occurred. The IHC technique detects the protease-resistant prion protein (PrP-res), which is considered a disease-specific marker for transmissible spongiform encephalopathies (TSE), regardless of the species affected. Of the 17 elk tested, 10 were positive by IHC. Only 2 of these 10 animals had shown clinical signs and histologic lesions of CWD, and an additional animal had histologic lesions only. The most consistently IHC-positive tissue was medulla oblongata, especially the obex. These results show that the PrP-res IHC test on brain tissue, specifically medulla oblongata at the obex, should be considered an essential component of any surveillance study intended to determine the incidence of CWD in captive or free-ranging cervids.     

Variable patterns of distribution of PrP(CWD) in the obex and cranial lymphoid tissues of Rocky Mountain elk (Cervus elaphus nelsoni) with subclinical chronic wasting disease

Sections of medulla oblongata, taken at the level of the obex, palatine tonsil and medial retropharyngeal lymph node from 10,269 captive Rocky Mountain elk (Cervus elaphus nelsoni), were examined by immunohistochemical staining with monoclonal antibody for the prion protein associated with the transmissible spongiform encephalopathy of cervids, chronic wasting disease (PrP(CWD)). The protein was detected in 226 of them. On the basis of the anatomical location of the deposits in the brainstem of 183 elk, four distinct patterns of distribution of PrP(CWD) within the parasympathetic region of the dorsal motor nucleus of the vagus nerve and the adjacent nuclei were observed. Mild gross lesions of chronic wasting disease (serous atrophy of fat) were observed in only three elk, all with spongiform degeneration; the other elk were considered to be in the preclinical stage of the disease. In contrast with the relatively predictable distribution of prion protein (PrP) in the brain and cranial nodes of sheep and mule deer, the distribution of PrP(CWD) in the brain and nodes of the elk was more variable and unrelated to their PrP genotype. One hundred and fifty-five of the 226 positive elk had deposits of PrP(CWD) in the brainstem and lymphoid tissues, 43 had deposits only in the lymphoid tissue and 28 had deposits only in the brainstem.     

Increased risk of chronic wasting disease in Rocky Mountain elk associated with decreased magnesium and increased manganese in brain tissue

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) of Rocky Mountain elk in North America. Recent studies suggest that tissue and blood mineral levels may be valuable in assessing TSE infection in sheep and cattle. The objectives of this study were to examine baseline levels of copper, manganese, magnesium, zinc, selenium, and molybdenum in the brains of Rocky Mountain elk with differing prion genotypes and to assess the association of mineral levels with CWD infection. Elk with leucine at prion position 132 had significantly lower magnesium levels than elk with 2 copies of methionine. Chronic wasting disease-positive elk had significantly lower magnesium than control elk. The incorporation of manganese levels in addition to magnesium significantly refined explanatory ability, even though manganese alone was not significantly associated with CWD. This study demonstrated that mineral analysis may provide an additional disease correlate for assessing CWD risk, particularly in conjunction with genotype.     

Chronic wasting disease in a Rocky Mountain elk

A 24-month-old Rocky Mountain elk was presented because of suspected chronic wasting disease (CWD). The animal was emaciated, had retained its winter hair, and had abnormal behavior patterns suggestive of CWD, including bruxism, ptyalism, and diminished flight zone size. Immunohistochemical analysis of the brain confirmed the diagnosis of CWD.     

北美地区鹿的慢性消耗性疾病

鹿慢性消耗性疾病(CWD)是鹿类动物的传染性海绵状脑病(TSE)。它是由朊病毒(PrP)引起的,临床主要表现为慢性型消耗,体重逐渐下降,行为异常,最后致死。该病主要感染北美地区的黑尾鹿、白尾鹿和美洲马鹿,一些野生和家养的反刍动物如牛、绵羊和山羊与染病鹿直接或间接接触也可被感染。目前还不能确定CWD与人和其它动物的TSE类疾病的关系。它的起源、发病机理、传播机制和途径尚不清楚。尽管还没有证据证明CWD可传染给人类,但是它对人类有潜在威胁。     

Detection of PrP(CWD) in postmortem rectal lymphoid tissues in Rocky Mountain elk (Cervus elaphus nelsoni) infected with chronic wasting disease.

Preclinical diagnostic tests for transmissible spongiform encephalopathies have been described for mule deer (Odocoileus hemionus), using biopsy tissues of palatine tonsil, and for sheep, using lymphoid tissues from palatine tonsil, third eyelid, and rectal mucosa. The utility of examining the rectal mucosal lymphoid tissues to detect chronic wasting disease (CWD) was investigated in Rocky Mountain elk (Cervus elaphus nelsoni), a species for which there is not a live-animal diagnostic test. Postmortem rectal mucosal sections were examined from 308 elk from two privately owned herds that were depopulated. The results of the postmortem rectal mucosal sections were compared to immunohistochemical staining of the brainstem, retropharyngeal lymph nodes, and palatine tonsil. Seven elk were found positive using the brainstem (dorsal motor nucleus of the vagus nerve), retropharyngeal lymph nodes, and palatine tonsil. Six of these elk were also found positive using postmortem rectal mucosal sections. The remaining 301 elk in which CWD-associated abnormal isoform of the prion protein (PrP(CWD)) was not detected in the brainstem and cranial lymphoid tissues were also found to be free of PrP(CWD) when postmortem rectal mucosal sections were examined. The use of rectal mucosal lymphoid tissues may be suitable for a live-animal diagnostic test as part of an integrated management strategy to limit CWD in elk.     

Development of monoclonal antibodies against the abnormal prion protein isoform (PrPres) associated with chronic wasting disease (CWD)

Monoclonal antibodies (mAbs) specific for the abnormal prion protein isoform (PrP^res) are indispensable for diagnosing chronic wasting disease (CWD). In this study, eight mAbs were developed by immunizing PrP knockout mice with recombinant elk PrP and an immunogenic PrP peptide. The reactivity of the mAbs to recombinant PrP and the PrP peptide was measured, and their isotypes were subsequently determined. Among them, four mAbs (B85-05, B85-08, B85-12, and B77-75) were shown by Western blotting to recognize proteinase K-treated brain homogenate derived from an elk suffering from CWD.     

A prion disease of cervids: chronic wasting disease

Chronic wasting disease (CWD) is a prion disease of deer, elk, and moose, initially recognized in Colorado mule deer. The discovery of CWD beyond the borders of Colorado and Wyoming, in Canada and as far east as New York, has led to its emergence as a prion disease of international importance. Epidemiological studies indicate that CWD is horizontally transmitted among free-ranging animals, potentially indirectly by prion-containing secreta or excreta contaminating the environment. Experimental CWD transmission attempts to other wild and domestic mammals and to transgenic mice expressing the prion protein of cattle, sheep, and humans have shed light on CWD species barriers. Transgenic mice expressing the cervid prion protein have proven useful for assessing the genetic influences of Prnp polymorphisms on CWD susceptibility. Accumulating evidence of CWD pathogenesis indicates that the misfolded prion protein or prion infectivity seems to be widely disseminated in many nonneural organs and in blood. This review highlights contemporary research findings in this prion disease of free-ranging wildlife.     

Preliminary observations of genetic susceptibility of elk (Cervus elaphus nelsoni) to chronic wasting disease by experimental oral inoculation.

To compare the genetic susceptibility of elk (Cervus elaphus nelsoni) with various alleles of the PRNP gene, which encodes the normal cellular prion protein, to chronic wasting disease (CWD), eight 8-month-old elk calves of 3 genotypes (2 132MM, 2 132LM, and 4 132LL) were orally dosed with CWD-infected brain material from elk. During postinoculation (PI) month 23, both 132MM elk had lost appetite, developed clinical signs of weight loss and central nervous system (CNS) dysfunction, and were euthanized. Two other elk (both 132LM) developed similar clinical signs of disease and were euthanized during PI month 40. All 4 affected elk had microscopic lesions of spongiform encephalopathy (SE), and PrPres, the disease-associated form of the prion protein, was detected in their CNS and lymphoid tissues by use of immunohistochemical (IHC) and Western blot (WB) techniques. These findings indicate that elk with MM and LM at codon 132 are susceptible to orally inoculated CWD. All 4 LL elk are alive at PI year 4 and are clinically normal, which suggests that 132LL elk may have reduced susceptibility to oral infection with CWD-infected material or may have prolonged incubation time.     

麋鹿重组朊蛋白单克隆抗体的制备及初步鉴定

为研制鹿慢性消耗性疾病(chronic wasting disease,CWD)单克隆抗体,本研究以原核表达后经纯化的麋鹿重组成熟朊蛋白(PrP^c)为免疫原免疫PrP^c基因敲除小鼠(PrP^c-null mice)。两次加强免疫后,利用淋巴细胞杂交瘤技术,取脾细胞与SP2/0骨髓瘤细胞进行细胞融合,间接ELISA方法筛选阳性杂交瘤细胞,采用3次有限稀释法实现杂交瘤细胞的亚克隆,筛选出5株能稳定分泌针对麋鹿重组成熟朊蛋白特异性单克隆杭体(McAbs)的杂交瘤细胞株5A5、3B2、6D12、5E3、1F5,其腹水ELISA效价均达到1∶10000以上。经鉴定,5株单抗均为IgG抗体,5A5、6D12、5E3株为IgG1亚类,3B2、1F5株为IgG2a亚类。Western blotting鉴定结果表明,获得的McAbs均能特异性识别麋鹿重组成熟朊蛋白和健康麋鹿脑组织匀浆中的PrP^c。本研究制备了CWD腹水McAbs,同时也为CWD的研究及其诊断方法的建立奠定了基础。     

Evaluation of Western blotting methods using samples with or without sodium phosphotungstic acid precipitation for diagnosis of scrapie and chronic wasting disease

The purpose of this study was to enhance the sensitivity of the Western blot (WB) test for use as an alternative and confirmatory method for the diagnosis of scrapie and chronic wasting disease (CWD) in Canada by comparing 2 sample preparation procedures: an abnormal prion protein (PrPSc) concentration procedure using sodium phosphotungstic acid (PTA) precipitation and a procedure using crude sample without precipitation. A total of 100 cerebrum samples (52 sheep and 48 elk), including 66 negative (31 sheep, 35 elk) and 34 positive (21 scrapie and 13 CWD positive) samples diagnosed by using immunohistochemistry (IHC) on retropharyngeal lymph node (RPLN) and medulla oblongata at obex, were tested by using WB with the 2 sample preparation procedures. The WB using non-PTA enriched sample (crude extract) detected, on average, only 71.7% (9 of 15, 60.0% for scrapie, 5 of 6, 83.3% for CWD) of the samples that tested positive by using WB with PTA enriched samples. No case was positive by WB using crude extract but negative by WB using PTA enriched sample. No false positive was found. Serial dilution of PTA precipitated samples demonstrated that the technique increases the detection limit approximately 100 fold. Additionally, the comparison of the WB and IHC on cerebrum from all the positive cases demonstrated that WB following PTA precipitation and IHC had 100% agreement by detecting 6 positive for CWD on cerebrum; while IHC detected scrapie in only 14 out of 15 positive cerebrum samples by using WB following PTA precipitation. Phosphotungstic acid precipitation is therefore a useful adjunct to WB analysis of scrapie and CWD and tissues.     

Risk Sensitivity and Hunter Perceptions of Chronic Wasting Disease Risk and Other Hunting,Wildlife, and Health Risks

This article examined relationships among hunter perceptions of personal health risks from chronic wasting disease (CWD), knowledge and information about CWD, and perceptions of other hunting, wildlife, and health risks. Data were obtained from surveys of 2,725 deer and elk hunters in Colorado. Cluster analysis grouped hunters into no (42%), slight (44%), and moderate (14%) risk groups based on perceptions of personal health risks from CWD (e.g., concern about health, become ill from CWD). There were minimal differences among groups in demographics, information sources, and knowledge about CWD. Hunters who perceived higher health risks from CWD (i.e., moderate risk), however, perceived greater risks associated with CWD to other humans, CWD to wildlife, hunting to personal health, other diseases to health, and the future of hunting. These findings illustrated the concept of risk sensitivity where hunters who perceived higher risks from CWD were predisposed to rate all other risks as large.     

Retrospective investigation of chronic wasting disease of cervids at the Toronto Zoo, 1973-2003

The occurrence of chronic wasting disease (CWD) at the Toronto Zoo was investigated retrospectively, based on an examination of management, animal health, and postmortem records, and immunohistochemical studies. Records of animal movements, clinical signs, and postmortem findings were examined for all cervids 1973-2003. All available samples of fixed, wax-embedded lymphoid or central nervous system tissue from cervids that died at the Toronto Zoo from 1973 to 2003, > 12 months of age, were tested, using prion protein immunostaining. Chronic wasting disease prion antigen was detected in 8 of 105 animals tested: 7 mule deer and 1 black-tailed deer. The most likely method of introduction was the importation of CWD-infected animals from a zoo in the United States. Animal-to-animal contact and environmental contamination were the most likely methods of spread of CWD at the zoo. No mule deer left the Toronto Zoo site, and the last animal with CWD died in 1981. Historic findings and ongoing testing of cervids indicate that the Toronto Zoo collection has very low risk of currently being infected with CWD.     

A retrospective study of the causes of morbidity and mortality in farmed elk (Cervus elaphus)

A survey of North American farmed elk (Cervus elaphus) producers was performed to determine the causes of sickness and mortality in farmed elk and to estimate mortality rates. Records over a 10-year period from 8 North American veterinary diagnostic pathology laboratories were also examined and summarized. The primary diagnosis for each record was used to classify diseases into categories such as parasitic, infectious, toxicological, and neoplastic. Nonspecific trauma was the most frequently reported known cause of mortality in both sexes and all age classes by elk producers. Ranked on perceived economic importance, producers cited trauma, chronic wasting disease, calf scours, dystocia, pneumonia, winter tick, tuberculosis, and grain overload. One-year mortality rates for adults and yearlings were 2.6% and 2.7%, respectively. Mortality rates for male and female adult animals were 2.4% and 2.7%, respectively. In general, the major findings of the survey matched reported causes for mortality provided by elk producers.