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Preclinical diagnosis of chronic wasting disease in captive mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus) using tonsillar biopsy.

The usefulness of tonsillar biopsy on live deer for preclinical diagnosis of the transmissible spongiform encephalopathy chronic wasting disease (CWD) was evaluated. Disease was tracked in a CWD-endemic herd using serial tonsillar biopsies collected at 6 to 9 month intervals from 34 captive mule deer (Odocoileus hemionus) and five white-tailed deer (O. virginianus). Tonsillar biopsies were examined for accumulation of PrP(CWD), the protein marker for infection, using immunohistochemical (IHC) staining. 26/34 (76%) mule deer and 4/5 (80%) white-tailed deer had PrP(CWD) accumulation in tonsillar biopsies; CWD was subsequently confirmed by post-mortem examination in all 30 of these tonsillar-positive deer. Six mule deer with IHC-negative tonsillar biopsies had positive brain and tonsillar IHC staining upon death 12 to 40 months following the last biopsy. PrP(CWD) accumulation in tonsillar biopsy was observed 2 to 20 months before CWD-related death and up to 14 months before onset of clinical signs of CWD. Tonsillar biopsies from 3-month-old mule deer (n=6) were IHC negative, but PrP(CWD) accumulation was detected in tonsillar biopsies from 7/10 mule deer by 19 months of age. Tonsillar biopsy evaluated with IHC staining is a useful technique for the preclinical diagnosis of CWD in live mule deer and white-tailed deer when intensive management approaches are possible.

Animals↗

Experimental infection of mule deer with Parelaphostrongylus tenuis.

Six adult and three fawn mule deer (Odocoileus hemionus) were experimentally infected with a range of 75-100 infective larvae of Parelaphostrongylus tenuis. Five of the six adult deer developed clinical signs of neurologic disease that terminated in paralysis between 35 and 80 days. The sixth deer developed slight signs of neurologic disease for 10 days, but recovered. All three mule deer fawns developed neurologic disease. Adult meningeal worms were recovered from the subdural space of the spinal cord of two fawns. Eggs were observed on the cranial dura mater of one of these fawns, indicating that P. tenuis can complete its life cycle provided mule deer can survive the damage resulting from the infection. Neither eggs nor larvae of P. tenuis were recovered from the feces or lungs of infected mule deer. Clinical signs and histologic lesions observed in experimentally infected mule deer resembled those reported in infected moose (Alces alces americana). Two critical periods were apparent in mule deer infected with P. tenuis: nematode migration through the spinal neural parenchyma, and penetration of the adult nematodes into the cranial neural parenchyma. While most adult deer were unable to survive the first critical period, fawns survived the first but succumbed to infection during the second critical period.

Animals↗

Isolation of bovine viral diarrhea virus from a free-ranging mule deer in Wyoming.

A noncytopathic type 1a bovine viral diarrhea virus (BVDV) was isolated from a free-ranging yearling female mule deer (Odocoileus hemionus) from northwestern Wyoming (USA). The mule deer was emaciated, weak, and salivating, and Arcanobacterium pyogenes was cultured from lung abscesses. Bovine viral diarrhea virus was isolated from lung, however, BVDV antigen was not detected by immunohistochemistry. The BVDV genotype was determined by reverse transcriptase polymerase chain reaction and the RNA sequences from the 5'UTR and E2 genes compared with sequences of a type 1a BVDV isolated from cattle from the same area as the deer. The sequences from the deer BVDV were distinct from those of the bovine type 1a BVDV, but similar to other bovine type 1a BVDVs. Seventy-four (60%) of 124 sera collected from mule deer in this area had serum neutralizing antibody titers to type 1a BVDV of > or = 1:32. The high prevalence of seropositive mule deer and isolation of BVDV suggests that this virus circulates in the mule deer population. The isolate described in this report is the second reported BVDV isolate from free-ranging deer in North America and the first from a mule deer.

Animal Diseases↗

Epidemiology of chronic wasting disease in captive white-tailed and mule deer.

The natural occurrence of chronic wasting disease (CWD) in a 1993 cohort of captive white-tailed deer (Odocoileus virginianus) afforded the opportunity to describe epidemic dynamics in this species and to compare dynamics with those seen in contemporary cohorts of captive mule deer (O. hemionus) also infected with CWD. The overall incidence of clinical CWD in white-tailed deer was 82% (nine of 11) among individuals that survived >15 mo. Affected white-tailed deer died or were killed because of terminal CWD at age 49-76 mo (x = 59.6 mo, SE = 3.9 mo). Epidemic dynamics of CWD in captive white-tailed deer were similar to dynamics in mule deer cohorts. Incidence of clinical CWD was 57% (4/7) among hand-raised (HR) and 67% (4/6) among dam-raised (DR) mule deer; affected HR mule deer succumbed at 64-86 mo of age (x = 72 mo; SE = 5 mo), and affected DR mule deer died at age 31-58 mo (x = 41.3 mo; SE = 6.1 mo). Sustained horizontal transmission of CWD most plausibly explained epidemic dynamics, but the original source of exposures could not be determined. Apparent differences in mean age at CWD-caused death among these cohorts may be attributable to differences in the timing or intensity of exposure to CWD, and these factors appear to be more likely to influence epidemic dynamics than species differences. It follows that CWD epidemic dynamics in sympatric, free-ranging white-tailed and mule deer sharing habitats in western North American ranges also may be similar.

Age Factors↗

Topographic distribution of scrapie amyloid-immunoreactive plaques in chronic wasting disease in captive mule deer (Odocoileus hemionus hemionus).

Chronic wasting disease (CWD), a progressive neurological disorder of captive mule deer, black-tailed deer, hybrids of mule deer and white-tailed deer and Rocky Mountain elk, is characterized neuropathologically by widespread spongiform change of the neuropil, intracytoplasmic vacuolation in neuronal perikarya and astrocytic hypertrophy and hyperplasia. We report the topographic distribution of amyloid plaques reactive to antibodies prepared against scrapie amyloid in CWD-affected captive mule deer (Odocoileus hemionus hemionus). Scrapie amyloid-immunoreactive plaques were found in the cerebral gray and white matter, in deep subcortical nuclei, in isolation or in clusters in areas of vacuolation, and perivascularly, in subpial and subependymal regions. In the cerebellum, immunoreactive amyloid plaques were observed in the molecular, pyramidal and granular layers. Scrapie amyloid-immunoreactive deposits were also seen in neuronal perikarya. Furthermore, amyloid plaques in CWD-affected captive mule deer were alcianophilic at 0.3 M magnesium chloride indicating the presence of weakly to moderately sulfated glycosaminoglycans. Our data corroborate that CWD in captive mule deer belongs to the subacute virus spongiform encephalopathies.

Alcian Blue↗

A cross-sectional study of epizootic lymphangitis in cart-mules in western Ethiopia.

A cross-sectional study was conducted to determine the prevalence of epizootic lymphangitis (EL) in 309 cart-mules (cart-pulling mules) in Bako and Ejaji towns, Western Ethiopia using clinical and microbiological examinations, between November 2002 and April 2003. The overall prevalence was 21% (CI=16.6-26%). The clinical, histological and mycological characteristics of EL in a cart-mule were similar with those in a horse. There was significant (chi2=133.5, P=0.001) association between tick infestation and EL lesions in study cart-mules. Amblyoma coherence and Boophilus genera were the ticks collected from lesions of cases of EL, and thus played a predisposing role. In conclusion, our results showed that EL has high prevalence in cart-mules in the two towns.

Animals↗

Testosterone concentrations, testes weight and morphology of mule drakes (Muscovy drake x Khaki Campbell).

1. Mule ducks were produced by naturally mating Muscovy drakes and Khaki Campbell ducks. 2. Semen was collected from 6-month-old mule drakes via an artificial vagina. The fluid was clear without any spermatozoa or spermatids. 3. Testes from 27-week-old mule drakes were smaller in size than those of Khaki Campbell drakes but heavier than Muscovy males of the same age. Histological sections of these testes revealed that spermatogenesis was not complete. 4. Testosterone concentration in the mule drakes was higher than in Muscovy males but similar to Khaki Campbell drakes. 5. Mule drakes have strong sexual drives as a result of high concentrations of testosterone, but, because spermatogenesis is incomplete, their semen had no sperm.

Aging↗

Characterization of a red blood cell antigen in donkeys and mules associated with neonatal isoerythrolysis.

A red cell antigen of donkeys and mules was identified using antibodies in serum from a mare which produced a mule foal affected with neonatal isoerythrolysis (NI). Subsequently antibodies with similar activity were identified in the sera of other mares which had produced mule foals and were produced by immunization of horses with blood from donkeys. The antigen detected by these antibodies does not correspond to any recognized horse red cell alloantigen. This may be a xenoantigen since all donkeys (and mules) tested have shared this antigen and all horses tested have lacked the antigen. The results suggest that all mule pregnancies (donkey sire x horse dam) are incompatible with regard to this factor and a potential for neonatal isoerythrolysis exists in all cases.

Animals↗

A fertile mule and hinny in China.

Anecdotal reports of fertility in female mules (jack donkey x mare) and hinnies (stallion x jenny donkey) have appeared in the literature over the years, but scientists have generally regarded them with scepticism. The fact that some of these hybrids can come into estrous and ovulate makes fertility conceivable, given that opportunity for mating arises. In China, where mules are bred extensively for work on the farms, a fertile female mule and a fertile female hinny have now been verified by chromosomal investigation. Each had mated with a donkey and produced a filly foal. The foals show unique hybrid karyotypes different from the mule's or hinny's and different from each other's. The studies make it clear that mule and hinny fertility, at least for the female hybrid, is a real possibility.

Animals↗

Prevalence of agglutinating antibodies to Toxoplasma gondii in adult and fetal mule deer (Odocoileus hemionus) from Nebraska.

Toxoplasma gondii is an apicomplexan parasite of mammals and birds. Herbivores acquire postnatal infection by ingesting oocysts from contaminated food or water. Toxoplasma gondii infection is common in white-tailed deer, Odocoileus virginianus, but little is known about the prevalence of infection in mule deer, O. hemionus. We examined sera from 89 mule deer from Nebraska for agglutinating antibodies to T. gondii using the modified direct agglutination test (MAT) with formalin-fixed tachyzoites as antigen. Thirty-one (35%) of the samples were positive at dilutions of > or = 1:25. Samples were examined from 29 fetuses from these mule deer and none were positive in the MAT. Sera from 14 white-tailed deer from Nebraska were also examined and 6 (43%) were positive for T. gondii. Samples were examined from 5 fetuses from these white-tailed deer and none was positive in the MAT. Our results in both deer species from Nebraska are similar to studies conducted in white-tailed deer from other regions of the United States. Our findings indicate that mule deer are frequently infected with T. gondii and that mule-deer meat may be a source of human infection.

Agglutination Tests↗

Sarcocystis hemionilatrantis (Sp. N.) life cycle in mule deer and coyotes.

Fifteen coyotes (Canis latrans) shed sporulated sporocysts in their feces after eating freshly ground skeletal muscles from a mule deer (Odocoileus hemionus hemionus) infected with microscopic-sized cysts of Sarcocystis. Sporocysts were shed intermittently from 12 to 36 days after ingestion of the infected meat. Sporocyst size averaged 14.4 X 9.3 mum. Eleven mule deer fawns orally inoculated with these sporocysts became infected and 9 of 11 died between post-inoculation days (PID) 27 and 63. Clinical signs of anorexia, weight loss, pyrexia and weakness were evident prior to death. A calf (Bos taurus) and two lambs (Ovis aries) orally inoculated with these sporocysts did not become infected and remained healthy throughout the experiments. Similarly, uninoculated control animals consisting of three mule deer fawns, two lambs and one calf remained healthy during the experiment. Preliminary histologic examinations conducted on selected tissues from all animals revealed microscopic-sized schizogonous stages in macrophages, between muscle fibers and near blood vessels in the esophagus, heart, biceps femoris, semi-membranosus, diaphragm and tongue from seven of eight fawns which died between PID 27 and 39. Developing or mature muscle cysts were not found in fawn tissue until PID 60. Sarcocysts were found in the three infected fawns examined after this time. Muscle cysts or earlier schizont stages were not found in tissues from the inoculated or uninoculated calves and lambs. A single muscle cyst was found in one control fawn; the other two control fawns were negative for both muscle cysts and other schizogonous stages. These results established that the life cycle of this species of Sarcocystis can be completed with coyotes as the definitive host and mule deer as the intermediate host. Based on the demonstrated host specificity and earlier findings, the name Sarcocystis hemionilatrantis is proposed for this parasite of mule deer and coyotes.

Animals↗

Experimental Fascioloides magna infections of mule deer (Odocoileus hemionus hemionus).

Six mule deer (Odocoileus hemionus hemionus) and one white-tailed deer (Odocoileus virginianus), approximately 5-mo-old, each were inoculated orally with 500 metacercariae of Fascioloides magna. All mule deer died from liver fluke infection between 69 and 134 days (mean = 114, SE = 9.9) after inoculation. Between 38 and 326 immature F. magna (mean = 102, SE = 45.5) were recovered from each deer at necropsy. Flukes were present in livers, lungs, and free in pleural and peritoneal spaces. Infection was characterized by necrotizing hepatitis, fibrosing peritonitis and pleuritis, and hematin pigment accumulation in liver, lung, and many other internal organs. Eggs of F. magna first were detected in feces of the white-tailed deer 28 wk after inoculation, and weekly thereafter until the healthy deer was euthanized at 31 wk. At necropsy, 205 F. magna, including 12 encapsulated mature and 193 nonencapsulated immature flukes were recovered from liver, lungs, and free in abdominal and thoracic spaces of the white-tailed deer. Based on these results, F. magna may be fatal to mule deer within 5 mo of infection. Like domestic sheep and goats, mule deer may be highly susceptible to infection, and it is unlikely mule deer can survive infection with large numbers of F. magna.

Animals↗

Mule deer (Odocoileus hemionus) and elk (Cervus elaphus) as experimental definitive hosts for Fascioloides magna.

In August 1992, six mule deer (Odocoileus hemionus hemionus) fawns and four elk (Cervus elaphus) calves (n = 2) or yearlings (n = 2) each were inoculated orally with 50, 250, or 2,000 metacercariae of the liver fluke Fascioloides magna to evaluate their potential to serve as definitive hosts. Animals were maintained for up to 403 days. Three mule deer each inoculated with 50 metacercariae survived the infection and shed eggs in feces; thus mule deer can function as definitive hosts for F. magna. The other three mule deer inoculated with 50 (n = 1) or 250 (n = 2) metacercariae died from fluke infection on days 91, 150, and 162 days postinoculation, respectively, and only immature F. magna were recovered. One elk calf inoculated with 2,000 metacercariae died from fluke infection 44 days after inoculation. The remaining three elk, each inoculated with 250 metacercariae, survived infection, and two of the three shed eggs in feces. The third elk contained only one immature F. magna at necropsy. The prepatent period in mule deer and elk was approximately 6 to 7 months.

Animals↗

Pathogen exposure patterns among sympatric populations of bighorn sheep, mule deer and cattle.

We sampled sympatric bighorn sheep (Ovis canadensis, n = 31), mule deer (Odocoileus hemionus, n = 38), and domestic cattle (n = 26) in the San Bernadino Mountains of southern California (USA) for the presence of Psoroptes spp. mites and for serologic evidence of exposure to bluetongue virus (BTV) and Babesia spp. From 1991 through 1994, Psoroptes spp. infestations were found on 12 (44%) of 27 bighorn sheep. No mites were found on mule deer or cattle. The BTV serum antibody prevalence in a cohort of 26 cattle ranged from 17 to 89%. There was no evidence of exposure to BTV in the bighorn sheep or mule deer. The cumulative serum antibody prevalence of Babesia spp. during the study was 35% in 26 bighorn sheep and 85% in 20 mule deer, while antibodies were not detected in a cohort of cattle when they were sampled in May (n = 23) and December (n = 22) of 1992. Based on these results, we concluded that infestation with Psoroptes spp. and exposure to BTV was limited to bighorn sheep and cattle, respectively. In contrast, Babesia spp. infections appeared to be common in both mule deer and bighorn sheep while there was no evidence of exposure in cattle.

Animals↗

Association of Moraxella ovis with keratoconjunctivitis in mule deer and moose in Wyoming.

Six cases of infectious keratoconjunctivitis (IKC) in mule deer (Odocoileus hemionus) and moose (Alces alces) in Wyoming (USA) were investigated during fall and winter of 1995 and 1996. Excessive lacrimation, mucopurulent conjunctivitis, keratitis, and corneal opacity were observed in mule deer. Moose had severe mucopurulent conjunctivitis, keratitis, and corneal ulceration. Hemolytic, non-piliated Moraxella ovis was isolated from two mule deer and two moose. We attempted to reproduce IKC in three mule deer fawns using an isolate of M. ovis from a clinically affected mule deer. These fawns did not develop clinical signs of infection and the bacterium was not reisolated from inoculated deer. Inoculated deer may not have developed clinical signs because deer were not exposed to ultraviolet light or mechanical insult before inoculation. In addition, the isolate used for inoculation may have lost virulence factors through passage, or M. ovis may not have been the primary pathogen responsible for clinical disease in the natural cases of IKC we investigated. The etiology of IKC in free-ranging wild ruminants remains poorly understood.

Animals↗

Relative vulnerability of chronic wasting disease infected mule deer to vehicle collisions.

We estimated chronic wasting disease (CWD) prevalence among vehicle-killed mule deer (Odocoileus hemionus) in select data analysis units (DAUs) in northern Colorado, USA, and compared these with estimated CWD prevalence among mule deer of the same sex sampled in the vicinity of collision sites to assess relative vulnerability of CWD-infected individuals to vehicle collisions. Twenty-five of 171 vehicle-killed mule deer tested positive for CWD (overall prevalence=0.146, 95% confidence interval [CI]=0.097-0.208); 173 of 2,317 deer sampled in the vicinity of these vehicle-killed deer tested positive (overall prevalence=0.075, 95% CI=0.064-0.085). In nine of ten DAU x sex comparisons, relative risk of CWD infection tended to be higher among vehicle-killed deer (range of estimated relative risks=1.6-15.9). Spongiform encephalopathy was detected in 12 of 20 (60%; 95% CI=39-81%) CWD-positive deer killed by vehicles and in 79 of 180 (44%; 95% CI=37-52%) CWD-positive deer detected via random sampling (relative risk=1.37; 95% CI=0.92-2.03), suggesting that infected deer killed by vehicles tended to be in later stages of disease than those killed by hunters. Our data offer evidence that CWD-infected mule deer may be relatively vulnerable to vehicle collisions. It follows that sampling of vehicle-killed mule deer may be exploited to increase efficiency of surveillance programs designed to detect new foci of CWD infection; moreover, evidence of increased susceptibility to vehicle collisions may aid in understanding vulnerability of CWD-infected individuals to other forms of death, particularly predation.

Accidents, Traffic↗

Cytogenetical analysis of the germ cell in the domestic drake and mule drake.

Experiments were conducted to study the germ cell of testes in the domestic drake (Tsaiya drake) and the mule drake. The histological sections of testes of both drakes were compared in this study. The results showed that rapid testicular growth did not occur until 7 weeks of age in domestic drakes and 15 weeks of age in the mule drakes. The rate of growth of the testes and the increase in the ratio of testes weight to body weight in domestic drakes at different ages were much more rapid than in the mule drakes. However, the opposite results were found in the body weights. In the mule drakes, spermatogonia and primary spermatocytes could be observed in the seminiferous tubules, but no secondary spermatocytes, spermatids or spermatozoa in seminiferous tubules were found up to 52 weeks of age. This showed that the meiotic division had not occurred in the mule drakes. In the domestic drakes, spermatogonia in the seminiferous tubules were observed at 1 week of age. Primary spermatocytes of some males appeared as early as the third week of age. Secondary spermatocytes and spermatids in the seminiferous tubules of some drakes began to appear at 5 weeks of age as a result of the meiotic division of the spermatocytes. At 7 weeks of age, spermatids were in process of metamorphosis, and spermatozoa were first observed in the seminiferous tubules of the drake. All testes of drakes 9 weeks of age contained spermatozoa. The percentage of the seminiferous tubules containing spermatozoa increased gradually with age, and approached a plateau at 15 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Serological and virological responses in mules and donkeys following inoculation with African horse sickness virus serotype 4.

Two groups, comprising 4 donkeys and 4 mules (group 1) and 4 donkeys and 3 mules (group 2), were used to determine the duration of viraemia and to monitor the development of antibodies following inoculation with African horse sickness virus (AHSV). One group of animals was given a single dose of attenuated AHSV serotype 4 (AHSV 4) vaccine. The second group was inoculated with a virulent field strain of AHSV 4. Both groups were subsequently challenged with the virulent field strain of AHSV 4, 51 and 58 days, respectively, after their primary inoculation. Blood and serum samples, collected on alternate days after the primary inoculations and also after subsequent challenge, were assayed for virus and antibodies. Seven of the 8 AHSV vaccinated (group 1) and 7 of the 7 AHSV inoculated (group 2) animals showed humoral antibody responses after primary inoculation. Although no infectious virus could be isolated from either group for the duration of the study, reverse transcription-PCR data obtained for the second group did show the presence of AHSV viral RNA from as early as day 5 in mules and day 9 in donkeys after the primary inoculation. Viral RNA was detected consistently up to day 47 in some animals and intermittently thereafter. There was no evidence of a second viraemia in any of the animals after challenge. The detection of specific antibodies, against AHSV 4 NS3 protein, in all animals confirmed that both donkeys and mules were infected and that the virus had replicated.

African Horse Sickness↗