Tolazoline-induced apnea in mule deer (Odocoileus hemionus)

Eighteen mule deer (Odocoileus hemionus) and six Columbia black-tailed deer (Odocoileus hemionus columbianus) were held in pens and repeatedly anesthetized from April 2004 through June 2005 as part of an external parasite study. Deer were anesthetized using a combination of Telazol and xylazine hydrochloride (HCL) administered intramuscularly. Tolazoline HCL was slowly administered at 4 mg/kg intravenously to reverse the effects of xylazine with good results. For 17 of the 19 mule deer anesthesias in the fall of 2004, a mean dose of 7.3 mg/kg of intravenous tolazoline (range 6.1-8.4 mg/kg) was given by mistake. This paper describes clinical signs of apnea, muscle tensing, and fasciculations immediately following intravenous administration of tolazoline HCL in mule deer (O. hemionus) at 1.5-3 times the recommended dose. Mean dose for black-tailed deer during this time was 8.1 mg/kg (range 5.5-12.4 mg/kg) with no clinical signs as seen in the mule deer. Based on these findings, intravenous tolazoline use in mule deer is recommended at < or = 4 mg/kg.     

Nonlethal experimental inoculation of Columbia black-tailed deer (Odocoileus hemionus columbianus) with virus of epizootic hemorrhagic deer disease.

Intramuscular or intravenous inoculation of 5 Columbia black-tailed deer (Odocoileus hemionus columbianus) with virus of epizootic hemorrhagic deer disease (EHD) did not produce overt clinical disease. Two white-tailed deer (Odocoileus virginianus) exposed identically died in 5 to 6 days. There were no significant lesions in 1 black-tailed deer euthanatized on postinoculation day 5. The EHd virus was not isolated from the spleen of that deer. Seroconversion occurred in black-tailed deer, from zero EHD virus antibody titer before inoculation to titers of 1:128 to 1:256 after inoculation.     

Mycobacteriosis in a black-tailed deer (Odocoileus hemionus columbianus) caused by Mycobacterium kansasii.

An eviscerated hunter-harvested female black-tailed deer (Odocoileus hemionus columbianus) was submitted to the Washington Department of Fish and Wildlife. The deer was emaciated, devoid of adipose tissue, and the parietal surface of the thoracic cavity contained multiple granulomas. Acid-fast bacteria were detected histologically from the granulomas and were isolated and identified as Mycobacterium kansasii, a nontuberculous mycobacterium sporadically reported to cause tuberculosis-like disease in a variety of vertebrates. This was the first report of symptomatic disease caused by M. kansasii in free-ranging deer. This case indicates that atypical mycobacteria can cause tuberculosis-like disease in free-ranging deer and illustrates the importance of identifying causative agents of tuberculosis-like disease in wildlife.     

An epizootic of adenovirus-induced hemorrhagic disease in captive black-tailed deer (Odocoileus hemionus).

Ten fawns and four adult black-tailed deer (Odocoileus hemionus) in a captive herd died as a result of adenovirus-induced hemorrhagic disease. Acute, systemic infections were characterized by hemorrhagic enteropathy, pulmonary edema, and occasional ulceration of the upper alimentary tract. Localized infections were limited to the upper alimentary tract and included stomatitis, pharyngitis, mandibular osteomyelitis, and rumenitis. In deer with acute, systemic infections, a diagnosis was made by histopathology and immunohistochemistry. The serum neutralization test was useful for confirming a diagnosis in animals with prolonged, localized infections. Deer originating from herds with a history of adenovirus infection should not be transferred to other captive herds or released into free-ranging populations because they may serve as carriers of adenovirus.     

Neospora caninum antibodies detected in Midwestern white-tailed deer (Odocoileus virginianus) by Western blot and ELISA

White-tailed deer (Odocoileus virginianus) serve to maintain the Neospora caninum life cycle in the wild. Sera from white-tailed deer from south central Wisconsin and southeastern Missouri, USA were tested for antibodies to N. caninum by Western blot analyses and two indirect ELISAs. Seroreactivity against N. caninum surface antigens was observed in 30 of 147 (20%) of WI deer and 11 of 23 (48%) of MO deer using Western blot analysis. Compared to Western blot, the two indirect ELISAs were found to be uninformative due to degradation of the field-collected samples. The results indicate the existence of N. caninum antibodies in MO and WI deer, and that Western blot is superior to ELISA for serologic testing when using degraded blood samples collected from deer carcasses.     

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.     

Serosurvey of antibodies against Toxoplasma gondii and Neospora caninum in white-tailed deer from Northern Mexico

The objective of this study was to determine the seroprevalence of Toxoplasma gondii and Neospora caninum in white-tailed deer from Northern Mexico. Sera from 532 white-tailed deer (Odocoileus virginianus) from three Northern states of Mexico were assayed for antibodies to T. gondii by ELISA and western blot. From these samples, 368 were available to test for N. caninum antibodies by ELISA. The overall prevalence for T. gondii antibodies was 13.9% (74/532; CI(95) 11-17) and for N. caninum 8.4% (31/368; CI(95) 6-12). There was a significant association between positive ELISA results for T. gondii, with management factors within ranches, such number of deer per hectare and geographic location of deer, but none for N. caninum. T. gondii infection in the deer from Guerrero, Coahuila had an increased risk than those from Nuevo Laredo, Tamaulipas (OR, 8.3; CI(95) 1.9-35.4; P<0.05) and ranches with one deer in 15ha had increased risk of positive association (OR, 2.61; CI(95) 1.5-4.4; P<0.05). These findings may have environmental or public health implications because venison can be an important meat source of T. gondii infections for humans and feral cats.     

Isolation of Neospora caninum from naturally infected white-tailed deer (Odocoileus virginianus)

Attempts were made to isolate Neospora caninum from naturally infected white-tailed deer (Odocoileus virginianus). A total of 110 deer killed during the 2003 hunting season in Virginia region were used for the isolation of N. caninum. Of these, brains from 28 deer that had NAT titer of 1:200 were inoculated into interferon-gamma gene knock out (KO) mice. N. caninum was isolated from the tissues of three deer and all three isolates were mildly virulent to KO mice. Only one of the isolates could be adapted to in vitro growth. Protozoa in the tissues of KO mice reacted with N. caninum-specific polyclonal antibodies and N. caninum DNA was demonstrated in infected tissues by PCR assays; sequences of portions of the ITS-1 and gene 5 loci were identical to those in the public database. This is the first record of in vitro isolation of N. caninum from white-tailed deer and lends credence to the white-tailed deer as an intermediate host for this parasite.     

Validation of monoclonal antibody F99/97.6.1 for immunohistochemical staining of brain and tonsil in mule deer (Odocoileus hemionus) with chronic wasting disease.

A new monoclonal antibody (MAb), F99/97.6.1, that has been used to demonstrate scrapie-associated prion protein PrP(Sc) in brain and lymphoid tissues of domestic sheep with scrapie was used in an immunohistochemistry assay for diagnosis of chronic wasting disease (CWD) in mule deer (Odocoileus hemionus). The MAb F99/97.6.1 immunohistochemistry assay was evaluated in brain and tonsil tissue from 100 mule deer that had spongiform encephalopathy compatible with CWD and from 1,050 mule deer outside the CWD-endemic area. This MAb demonstrated abnormal protease-resistant prion protein (PrP(res)) in brains of all of the 100 mule deer and in 99 of the 100 tonsil samples. No immunostaining was seen in samples collected from deer outside the endemic area. MAb F99/97.6.1 demonstrated excellent properties for detection of PrP(res) in fresh, frozen, or mildly to moderately autolytic samples of brain and tonsil. This immunohistochemistry assay is a sensitive, specific, readily standardized diagnostic test for CWD in deer.     

Comparison of histological lesions and immunohistochemical staining of proteinase-resistant prion protein in a naturally occurring spongiform encephalopathy of free-ranging mule deer (Odocoileus hemionus) with those of chronic wasting disease of captive mule deer

In this investigation, the nature and distribution of histologic lesions and immunohistochemical staining (IHC) of a proteinase-resistant prion protein were compared in free-ranging mule deer (Odocoileus hemionus) dying of a naturally occurring spongiform encephalopathy (SE) and captive mule deer dying of chronic wasting disease (CWD). Sixteen free-ranging deer with SE, 12 free-ranging deer without SE, and 10 captive deer with CWD were examined at necropsy. Tissue sections were stained with hematoxylin and eosin, and duplicate sections were stained with a monoclonal antibody (F89/160.1.5). Histological lesions in the free-ranging deer with SE and captive deer with CWD were found throughout the brain and spinal cord but were especially prominent in the myelencephalon, diencephalon, and rhinencephalon. The lesions were characterized by spongiform degeneration of gray matter neuropil, intracytoplasmic vacuolation and degeneration of neurons, and astrocytosis. IHC was found throughout the brain and retina of deer with SE and CWD. Positive IHC was found in lymphoid tissue of deer with SE and CWD. Histologic lesions and IHC were not found in multiple sections of integument, digestive, respiratory, cardiovascular, endocrine, musculoskeletal, and urogenital systems of deer with SE or CWD. Comparison of histologic lesions and IHC in tissues of free-ranging deer with those of captive deer provides strong evidence that these two diseases are indistinguishable morphologically.     

Effects of starch and fibre in pelleted diets on nutritional status of mule deer (Odocoileus hemionus) fawns

To compare the effects of a low-starch, high-fibre diet [LSHF; 51.6% neutral detergent fibre (NDF), 3.0% starch, 14.8% crude protein (CP)] and a high-starch, low-fibre diet (HSLF; 33.3% NDF, 20.0% starch, 19.6% CP) on the nutritional status of captive exotic ruminants, 16 mule deer (Odocoileus hemionus) were fed one of these two diets ad libitum with ≤25% alfalfa hay cubes from 10 days to 68 weeks of age. During five sampling periods beginning in November and spaced 6-12 weeks apart thereafter, feed intake, rumen and blood chemistry, faecal scores, growth and body condition were measured. Dry matter intake, digestible energy intake, time spent ruminating and feeding and blood acetate concentration were greater for deer fed LSHF (p < 0.05 for all). Lower dietary CP led to reduced blood urea nitrogen for deer consuming LSHF (p = 0.004). Deer had the same faecal scores, growth and body fat among treatments (all p > 0.05). These findings show pelleted diets with less starch, more fibre, and reduced protein met the energy and protein requirements of growing mule deer, a medium-sized browsing ruminant, as well as traditional grain-based diets while more closely mimicking natural forages and stimulating a more natural feeding behaviour.     

Serum biochemical and electrophoretic values from four deer species and from pronghorn antelope.

Serums from 4 species of deer and 1 species of antelope were analyzed for various components in order to define an animal disease model for sickle cell disease in people. Animal species included black-tailed deer (Odocoileus hemionus columbianus), mule deer (Odocoileus hemionus), sika deer (Cervus nippon nippon), fallow deer (Dama dama), and pronghorn antelope (Antilocapra americana). The mean serum values for total bilirubin, total protein, albumin, creatinine, urea nitrogen, and electrolytes were similar in all species and were in the normal range for human beings. Cholesterol and uric acid values for all animals were lower than those for people. Alkaline phosphatase values in the 4 cervid species were higher than in the pronghorn antelope. Values for glutamic oxalacetic transaminase were lower in the cervids than in the pronghorn antelope. Lactic dehydrogenase values were similar in the 5 species. High activities for glutamic oxalacetic transaminase and lactic dehydrogenase in the 5 species probably related to muscle mass and great muscular activity.     

Field validation and assessment of an enzyme-linked immunosorbent assay for detecting chronic wasting disease in mule deer (Odocoileus hemionus), white-tailed deer (Odocoileus virginianus), and Rocky Mountain elk (Cervus elaphus nelsoni).

Tissue samples (n = 25,050 total) from 23,256 mule deer (Odocoileus hemionus), Rocky Mountain elk (Cervus elaphus nelsoni), and white-tailed deer (Odocoileus virginianus) collected statewide in Colorado were examined for chronic wasting disease (CWD) using an enzyme-linked immunosorbent assay developed by Bio-Rad Laboratories, Inc. (brELISA), in a 2-phase study. In the validation phase of this study, a total of 4,175 retropharyngeal lymph nodes (RLN) or obex (OB) tissue samples were examined independently by brELISA and immunohistochemistry (IHC). There were 137 IHC-positive samples and 4,038 IHC-negative samples. Optical density (OD) values from brELISA were classified as "not detected" or "suspect" based on recommended cutoff values during the validation phase. Using IHC-positive cases as known CWD-infected individuals and assuming IHC-negative cases as uninfected, the relative sensitivity of brELISA depending on species ranged from 98.3% to 100% for RLN samples and 92.1% to 93.3% for OB samples; the relative specificity of brELISA depending on species ranged from 99.9% to 100% for RLN samples and was 100% for OB samples. Overall agreement between brELISA and IHC was > or = 97.6% in RLN samples and > or = 95.7% in OB samples of all species where values could be calculated; moreover, mean brELISA OD values were > or = 46X higher in IHC-positive samples than in IHC-negative samples. Discrepancies were observed only in early-stage cases of CWD. Based on the validation phase data, only RLN samples were collected for the field application phase of this study and only samples with brELISA OD values > 0.1 were examined by IHC. Among 20,875 RLN samples screened with brELISA during this second testing phase, 155 of 8,877 mule deer, 33 of 11,731 elk, and 9 of 267 white-tailed deer samples (197 total) had OD values > 0.1 and were further evaluated by IHC to confirm evidence of CWD infection. Of cases flagged for IHC follow-up, 143 of 155 mule deer, 29 of 33 elk, and all 9 white-tailed deer were confirmed positive. Mean (+/- SE) OD values for IHC-positive cases detected during the field application phase were comparable with those measured in RLN tissues during the validation phase. Based on these data, brELISA was determined to be an excellent rapid test for screening large numbers of samples in surveys designed to detect CWD infections in deer and elk populations.     

Lesions caused by Parelaphostrongylus odocoilei (Nematoda: Metastronglyloidea) in two cervid hosts

Pathologic effects and host response were evaluated in seven white-tailed deer (Odocoileus virginianus) and six mule deer (O. hemionus hemionus) each exposed per os to 300 or 1000 third-stage larvae of Parelaphostrongylus odocoilei. Pathologic effects in mule deer consisted of hemorrhagic myositis throughout skeletal muscles, severe verminous pneumonia, and moderate lymphadenitis. The major host response was a granulomatous inflammation associated with nematode eggs and larvae. Granulomas obliterated the normal architecture of affected tissues. Pathologic effects and host response were minimal in white-tailed deer. P. odocoilei is considered a potential direct or indirect pathogen in mule deer but an insignificant parasite in white-tailed deer.     

Distribution of protease-resistant prion protein and spongiform encephalopathy in free-ranging mule deer (Odocoileus hemionus) with chronic wasting disease

Serial sections of brain and palatine tonsil were examined by immunohistochemical staining (IHC) using monoclonal antibody F89/160.1.5 for detecting protease-resistant prion protein (PrP(res)) in 35 hunter-killed mule deer (Odocoileus hemionus) with chronic wasting disease. Serial sections of brain were stained with hematoxylin and eosin and examined for spongiform encephalopathy (SE). Clinical signs of disease were not observed in any of these deer. On the basis of the location and abundance of IHC and the location and severity of SE, deer were placed into four categories. Category 1 (n = 8) was characterized by IHC in the palatine tonsil with no evidence of IHC or SE in the brain. Category 2 (n = 13) was characterized by IHC in the palatine tonsil and IHC with or without SE in the dorsal motor nucleus of the vagus nerve (DMNV). Category 3 (n = 2) was characterized by IHC in the palatine tonsil, IHC with SE in the myelencephalon, and IHC without SE in the hypothalamus. Category 4 (n = 12) was characterized by IHC in the palatine tonsil and IHC with SE throughout the brain. Category I may represent early lymphoid tissue localization of PrP(res). The DMNV appears to be the most consistent single neuroanatomic site of detectable PrP(res). Categories 2-4 may represent a progression of spread of PrP(res) and SE throughout the brain. IHC in tonsil and brain and SE in brain were not detected in 208 control deer.     

Dermatomycosis (Ringworm) in Mule Deer (Odocoileus hemionus)

Six mule deer (Odocoileus hemionus) with dermatomycosis are described. Trichophyton verrucosum was isolated from four. All infections were mild and were not debilitating. The lesions involved the legs in five animals and the face in two. This is the second report of ringworm in a wild ungulate in North America.     

Lesions and transmission of experimental adenovirus hemorrhagic disease in black-tailed deer fawns

Adenovirus infection was the cause of an epizootic of hemorrhagic disease that is believed to have killed thousands of mule deer (Odocoileus hemionus) in California during the latter half of 1993. A systemic vasculitis with pulmonary edema and hemorrhagic enteropathy or a localized vasculitis associated with necrotizing stomatitis/pharyngitis/glossitis or osteomyelitis of the jaw were common necropsy findings in animals that died during this epizootic. To study transmission of adenovirus infection in deer and susceptibility of black-tailed deer (Odocoileus hemionus columbianus) fawns to adenovirus infection, six 3-6-month-old black-tailed fawns were divided into two treatment groups. One group was inoculated intravenously and the other group was inoculated through the mucous membranes of the eyes, nose and mouth with purified adenovirus. Each treatment group also included two additional fawns (four total) that were not inoculated but were exposed to inoculated animals (contact animals). One fawn served as a negative control. Between 4 and 16 days postinoculation, 8/10 fawns developed systemic or localized infection with lesions identical to lesions seen in animals with natural disease that died during the epizootic. Transmission was by direct contact, and the route of inoculation did not affect the incubation period or the distribution of the virus (systemic or the localized infection). Immunohistochemical analysis using polyclonal antiserum against bovine adenovirus type 5 demonstrated staining in endothelial cells of vessels in numerous tissues in animals with systemic infection and endothelial staining only in vessels subtending necrotic foci in the upper alimentary tract in animals with the localized form of the disease. All inoculated or exposed animals had staining in the tonsillar epithelium. Transmission electron microscopic examination of lung and ileum from two fawns with pulmonary edema and hemorrhagic enteropathy demonstrated endothelial necrosis and adenovirus virions in endothelial cell nuclei. Adenovirus was reisolated in black-tailed deer pulmonary artery endothelial cells using lung homogenate of the first fawn that developed systemic adenovirus infection. Serum virus neutralization test results suggest that this deer adenovirus is a new serotype.     

Seroepidemiology of Neospora caninum and Toxoplasma gondii infection in yaks (Bos grunniens) in Qinghai, China

Neospora caninum is a protozoan parasite and is closely related to Toxoplasma gondii, but they are antigenically different. N. caninum and T. gondii infection in a variety of animals such as cattle, dogs, and cats has been reported, but there is little information on the infection of these parasites in domestic yaks. Seroprevalence of antibodies to T. gondii and N. caninum in yaks (Bos grunniens) from eight regions of Qinghai, China were investigated by the indirect agglutination test (IAT) and ELISA, respectively. A total of 112 (11.8%) of 946 serum samples were positive for antibodies to T. gondii, and 21 samples (2.2%) were positive to N. caninum. Two of the yaks had antibodies to both parasites. There was no apparent association of T. gondii infection with age of the animals. The results indicate that T. gondii infection is prevalent in Chinese yaks in most parts of Qinghai province and N. caninum infection rate in the same species is relatively low. This is the first large study showing the infection of T. gondii and N. caninum in domestic yaks.     

Serologic profile of exotic deer at Point Reyes National Seashore

Serotests were conducted on axis (Axis axis) and fallow (Dama dama) deer at Point Reyes National Seashore to determine their status with respect to nine diseases enzootic to the native black-tailed deer (Odocoileus hemionus columbianus) or to resident dairy cattle. In the exotic deer, the proportion of animals that were seropositive included: anaplasmosis, 35%; bluetongue, 48%; brucellosis, 0%; bovine viral diarrhea, 2%; infectious bovine rhinotracheitis, 3%; leptospirosis, 7%; parainfluenza-3, 49%; toxoplasmosis, 8%; and Q fever, 51%. The prevalence of antibodies among a small sample of the black-tailed deer included anaplasmosis, 100%; toxoplasmosis, 29%; and Q fever, 57%. The antibody prevalences in a sample of dairy cattle in the area included anaplasmosis, 19%; toxoplasmosis, 8%; and Q fever, 100%.     

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.