Testing whether adrenal steroids mediate phenotypic and physiologic effects of elevated salinity on larval tiger salamanders

Salinity (sodium chloride, NaCl) from anthropogenic sources is a persistent contaminant that negatively affects freshwater taxa. Amphibians can be susceptible to salinity, but some species are innately or adaptively tolerant. Physiological mechanisms mediating tolerance to salinity are still unclear, but changes in osmoregulatory hormones such as corticosterone (CORT) and aldosterone (ALDO) are prime candidates. We exposed larval barred tiger salamanders (Ambystoma mavortium) to environmentally relevant NaCl treatments (<32–4000 mg·L⁻¹) for 24 days to test effects on growth, survival, and waterborne CORT responses. Of those sampled, we also quantified waterborne ALDO from a subset. Using a glucocorticoid antagonist (RU486), we also experimentally suppressed CORT signaling of some larvae to determine if CORT mediates effects of salinity. There were no strong differences in survival among salinity treatments, but salinity reduced dry mass, snout–vent length, and body condition while increasing water content of larvae. High survival and sublethal effects demonstrated that salamanders were physiologically challenged but could tolerate the experimental concentrations. CORT signaling did not attenuate sublethal effects of salinity. Baseline and stress-induced (after an acute stressor, shaking) CORT were not influenced by salinity. ALDO was correlated with baseline CORT, suggesting it could be difficult to decouple the roles of CORT and ALDO. Future studies comparing ALDO and CORT responses of adaptively tolerant and previously unexposed populations could be beneficial to understand the roles of these hormones in tolerance to salinity. Nevertheless, our study enhances our understanding of the roles of corticosteroid hormones in mediating effects of a prominent anthropogenic stressor.     

Blood culture and stimulation conditions for the diagnosis of tuberculosis in cervids by the Cervigam assay

Mitogen- and antigen-induced interferon-gamma (IFN-gamma) responses of peripheral blood leucocytes from cervids were evaluated by a commercial whole-blood assay. The assay was applied to Mycobacterium bovis-infected white-tailed deer and reindeer, M bovis BCG-vaccinated white-tailed deer and elk, and unvaccinated, uninfected white-tailed deer, fallow deer, elk and reindeer. The responses of the M bovis-infected white-tailed deer to pokeweed mitogen (PWM) varied with time and between individuals. The responses of the M bovis-infected reindeer to PWM and M bovis purified protein derivative (PPD) were positively associated. Samples from tuberculosis-free captive herds in various parts of the USA were also evaluated. Four per cent of fallow deer, 20 per cent of elk, 44 per cent of white-tailed deer, and 91 per cent of reindeer had responses to PWM exceeding 0.25 Delta optical density, that is, PWM stimulation minus no stimulation. The specificity of the responses to M bovis PPD and a Mycobacterium tuberculosis complex-specific antigen rESAT-6:CFP-10, excluding animals not responding to PWM, ranged from 78 per cent to 100 per cent and was dependent upon the species and the positive response cut-off value. The results show that the commercial assay is valid for the detection of TB in reindeer; however, further development of the assay will be required before it is used in surveillance programmes for white-tailed deer, fallow deer, and elk.     

Evaluation of historical factors influencing the occurrence and distribution of Mycobacterium bovis infection among wildlife in Michigan

OBJECTIVES: To determine historical events leading to establishment of bovine tuberculosis in the white-tailed deer population in the northeastern corner of the lower peninsula (NELP) of Michigan and describe factors relevant to the present outbreak of bovine tuberculosis in Michigan. SAMPLE POPULATION: Cattle and white-tailed deer in Michigan from 1920 to 1990. PROCEDURES: A search of extant historical documents (eg, scientific journals, books, public reports, and correspondence and internal reports from governmental agencies) was conducted. Factors investigated included the number of cattle and prevalence of tuberculosis, deer population and density levels, and changes in regional environments affecting the population and management of cattle and wild deer. RESULTS: High deer numbers and severe winter feed shortages resulting from habitat destruction in the NELP in 1930 contributed to the transmission of tuberculosis from cattle to deer. Starvation increased the susceptibility of deer to infection and modified behavior such that exposure to infected cattle was increased. Relocation of deer resulted in spread of infection to other sites, including locations at which spatial clusters of tuberculosis presently exist. Ribotyping of Mycobacterium bovis from a human patient suggests that the strain of M. bovis presently infecting white-tailed deer in the region is the same strain that affected cattle farms at that time. CONCLUSIONS AND CLINICAL RELEVANCE: Feeding deer to maintain numbers above the normal carrying capacity of the NELP led to deer depending on consumption of livestock feed for survival during winter and increased contact with domestic cattle. This practice should be avoided.     

Spatial analysis of Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus) in Michigan, USA

The wild white-tailed deer (Odocoileus virginianus) population in Michigan, USA, has endemic Mycobacterium bovis. We determined whether there were spatial clusters of retrospective TB cases in white-tailed deer in northeastern Michigan and identified specific factors associated with the spatial clusters. Data from hunter-harvested deer (age, gender, TB status, and geographic section) were collected by the Michigan Department of Natural Resources (MDNR) during TB surveillance from 1995 to 2002. Land cover (vegetation, land-use) and land type (soil types and drainage characteristics, landforms) described potential deer habitats. Specific locations of large-scale supplemental feeding sites were collected from the MDNR aerial surveillance program from 1997 to 2002. Analyses were conducted using principal components derived from environmental data (and other risk factors) on spatial clusters of disease (identified by the spatial scan statistic). Spatial effects were incorporated into the multivariable analyses by using a neighborhood approach. A total of 420 deer with M. bovis infection were identified from 1995 to 2002, out of 39,451 harvested deer from 3216 TRS units, and spatial clusters of cases were identified. A total of seven principal components of environmental data were generated. Clusters were associated with the presence of large expanses of deciduous forests on moraine ridges separated by low areas of forested wetlands, and the presence of many small lakes. Factors that promoted congregation of deer for extended periods of time (natural cover, access to water, and less human contact) appeared to be associated with increased odds of TB positivity. This suggests that there are specific areas where interventions can be implemented to reduce congregation of animals and disrupt the cycle of infection transmission.     

Targeting the detection of chronic wasting disease using the hunter harvest during early phases of an outbreak in Saskatchewan, Canada

Chronic wasting disease (CWD) is a fatal disease of North American cervids that was first detected in a wild, hunter-shot deer in Saskatchewan along the border with Alberta in Canada in 2000. Spatially explicit models for assessing factors affecting disease detection are needed to guide surveillance and control programs. Spatio-temporal patterns in CWD prevalence can be complicated by variation in individual infection probability and sampling biases. We assessed hunter harvest data of mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus) during the early phases of an outbreak in Saskatchewan (i.e., 2002-2007) for targeting the detection of CWD by defining (1) where to look, and (2) how much effort to use. First, we accounted for known demographic heterogeneities in infection to model the probability, P(E), that a harvested deer was infected with CWD given characteristics of the harvest location. Second, in areas where infected deer were harvested we modelled the probability, P(D), of the hunter harvest re-detecting CWD within sample units of varying size (9-54 km(2)) given the demographics of harvested deer and time since first detection in the study area. Heterogeneities in host infection were consistent with those reported elsewhere: mule deer 3.7 times >white-tailed deer, males 1.8 times>females, and quadratically related to age in both sexes. P(E) increased with number of years since the first detection in our study area (2002) and proximity to known disease sources, and also varied with distance to the South Saskatchewan River and small creek drainages, terrain ruggedness, and extent of agriculture lands within a 3 km radius of the harvest. The majority (75%) of new CWD-positive deer from our sample were found within 20 km of infected deer harvested in the previous year, while approximately 10% were greater than 40 km. P(D) modelled at 18 km(2) was best supported, but for all scales, P(D) depended on the number of harvested deer and time since the first infected deer was harvested. Within an 18 km(2) sampling unit, there was an 80% probability of detecting a CWD-positive deer with 16 harvested deer five years after the initial infected harvest. Identifying where and how much to sample to detect CWD can improve targeted surveillance programs early in the outbreak of the disease when based on hunter harvest.     

Evaluating use of cattle winter feeding areas by elk and white-tailed deer: Implications for managing bovine tuberculosis transmission risk from the ground up

Transmission of bovine tuberculosis (Mycobacterium bovis) among wildlife and livestock has created important risks for conservation and agriculture. Management strategies aimed at controlling TB have typically been top-down, regionally focused, and government-led programs that were at best only partially successful. The purpose of this study was to quantify co-mingling of elk and white-tailed deer (WTD) with cattle at multiple spatial scales (i.e., the regional farm scale and winter cattle feeding area patch) in southwestern Manitoba, Canada, to assess the potential for bovine tuberculosis transmission and identify alternative management strategies. For each spatial scale we quantified use of cattle farms by elk and white-tailed deer. We mailed questionnaires to rural households and then conducted personal interviews with 86 cattle farmers to map the spatial distribution of their cattle winter feeding areas at a fine scale. We deployed Global Positioning System (GPS) collars on 48 wild elk and 16 wild white-tailed deer from 2003 to 2011. Elk were observed on farms by 66% of cattle producers, including 5% and 20% who observed direct and indirect contact, respectively, between elk and cattle. Cattle producers consistently (≈100%) observed white-tailed deer on their farms, including 11% and 47% whom observed direct and indirect contact, respectively, between white-tailed deer and cattle. A higher probability of white-tailed deer–cattle contact at the regional scale occurs on farms that (1) left crop residues specifically for wildlife, (2) had larger cattle herds, (3) used round bale feeders, and (4) were farther away from protected areas. None of the GPS-collared elk locations overlapped with cattle winter feeding areas. In contrast, 21% of GPS-collared white-tailed deer locations overlapped with winter cattle winter feeding areas (22% of these were from male WTD and 78% were from female WTD). White-tailed deer selected cattle winter feeding areas with higher (1) forage crop, (2) grassland/rangeland, and (3) forest cover around the cattle feeding area. Farmers overall expressed strongly negative attitudes toward eradicating the elk population or fencing the park to eradicate TB, but were generally supportive of less invasive and farm-based approaches. Our results suggested that management efforts to prevent TB transmission at the wildlife-agriculture interface can be effectively implemented using a ‘bottom-up’ approach that focuses on practical, farm-based mitigation strategies. This approach can be implemented by individual farm operators, is relatively low cost, and is generally well supported by farmers relative to other more extreme and controversial measures like wildlife eradication.     

Epizootic hemorrhagic disease virus in Montana: isolation and serologic survey

Epizootic hemorrhagic disease virus (EHDV) was isolated in Vero cell culture from the spleen and whole blood of a white-tailed deer (Odocoileus virginianus). A 10% spleen suspension caused acute hemorrhagic disease (HD) when inoculated into an experimental white-tailed deer and resulted in the recovery of EHDV from the blood of the experimental animal at 5 days after inoculation. The virus was identified as EHDV serotype 2 through indirect fluorescent antibody tests, electron microscopy, and reciprocal cross-neutralization tests. Approximately 73% (36/49) of the mule deer, 5% (2/42) of the white-tailed deer, and 79% (249/314) of the cattle samples tested from areas where HD had been reported were EHDV seropositive. Although none of the white-tailed deer was bluetongue virus seropositive, 29% of the mule deer and 3% of the cattle tested from "active" HD areas possessed bluetongue virus precipitating antibody.     

Management of indigenous North American deer at the end of the 20th century in relation to large predators and primary production

Five deer species occupy North America: caribou (3.6 x 10(6) individuals), moose (1.1 x 10(6)), white-tailed deer (28.5 x 10(6)), mule deer (5.0 x 10(6)) and wapiti (1.1 x 10(6)). Caribou characterise the north of the boreal forest and the tundra, whereas moose dominate in coniferous and mixed forests growing further south. White-tailed deer are typical of the deciduous forests of the east while mule deer replace them in the mountainous terrain of the west. Wapiti possess the smallest range, mostly adjacent to the prairies to the west. The two large obligate carnivores preying on deer show a reduced distribution: wolves are almost restricted to Canada, and cougar to the mule deer range. We determined the current status of each species with the help of a questionnaire mailed to all jurisdictions harbouring deer. Most reports of threatened populations concerned caribou whereas many jurisdictions declared overabundance of white-tailed deer and wapiti. Hunting was allowed for all species when they abounded in a jurisdiction. Hunters harvested annually 7.0 x 10(6) deer on the continent, 87% being white-tailed deer. The two species that caused most conflicts with humans had the highest harvest rate: 16-17%. In terms of biomass, white-tailed deer and wapiti yielded the highest harvests, with 55 and 39 kg x km-2 of range, respectively. The average standing biomass of deer in winter ranged between 28 kg x km-2 in Nevada to 901 kg x km-2 in Indiana. The lowest standing biomasses occurred in the boreal forest (predators), in the prairies (agriculture) and in the south-west (aridity), and the highest ones in the south-east, where only white-tailed deer is present. The current abundance of deer in North America parallels, in general, the primary production of the landscape (r2 = 0.38; P < 0.0001), but predators and human activity modify this pattern.     

A cross-sectional study of reproductive indices and fawn mortality in farmed white-tailed deer

Data were obtained from a questionnaire administered to a random sample of Canadian and United States white-tailed deer (WTD) farmers. Reproductive indices and survival of fawns from birth until 1 y of age were examined. Major factors in limiting herd increase were a low reproductive rate (88 fawns per 100 does exposed to bucks) and a 30% mortality of fawns from birth until 1 y of age. The latter figure differs from reported mortality rates in fallow deer and red deer/wapiti. The unacceptably high neonatal mortality on WTD farms was determined to be as important to herd productivity as failure to produce a live fawn. Industry wide, "benchmark" estimates of reproductive performance, mortality rates, and productivity are provided, allowing farmers to compare their herds against these "benchmarks" to identify areas needing improvement.     

Novel hemotropic Mycoplasma species in white-tailed deer (Odocoileus virginianus)

Globally, hemotropic Mycoplasma spp. are emerging or re-emerging zoonotic pathogens that affect livestock, wildlife, companion animals, and humans, potentially causing serious and economically important disease problems. Little is known about hemotropic Mycoplasma spp. prevalence, host-specificity, or route of transmission in most species, including wildlife. DNA amplification by PCR targeting the 16SrRNA and the RNaseP genes was used to establish the presence and prevalence of hemotropic Mycoplasma spp. in a white-tailed deer (O. virginianus) population in eastern North Carolina. Sixty-five deer (89%) tested positive for hemotropic Mycoplasma spp. where sequence analysis of the 16SsRNA and the RNaseP genes indicated the presence of at least three distinct species. This study represents the first detection of three distinct hemotropic Mycoplasma species in white-tailed deer and the first report of two novel hemotropic Mycoplasma species.     

Use of adrenocorticotropin challenges to indicate chronic stress responses of laying hens in several housing alternatives

The use of adrenocorticotropin (ACTH) challenges was tested as an indicator of the level of physiological stress response of laying hens in several housing alternatives. Single Comb White Leghorn (SCWL) hens were challenged with intravenous ACTH injections (either .33, 1.0 or 3.0 IU/kg body weight). Plasma corticosterone (CORT) concentrations taken at intervals from 10 to 180 min following ACTH challenges revealed CORT response curves which peaked within 30 min at 34-, 54- and 60-fold increases over pre-challenge CORT levels with doses of .33, 1.0 and 3.0 IU/kg ACTH, respectively. Peak CORT levels for 1.0 and 3.0 IU/kg of ACTH did not differ; however, CORT levels for 1.0 IU/kg ACTH were significantly elevated above the peak for .33 IU/kg ACTH. Laying hens housed in cage and floor management alternatives were challenged with .33 and 1.0 IU/kg of ACTH after 10 months of lay. Cage management treatments were 3, 4 or 5 hens per cage and floor treatments with hens kept at 2 densities. Significant differences in the ACTH-induced CORT response between management alternatives could be detected following injection of .33 IU/kg ACTH, but not with the higher ACTH dose (1.0 IU/kg). These studies demonstrated that the CORT response to the lower dose of ACTH seemed to be more effective in assessing management differences, but further clarification may be necessary before ACTH challenges can be accurately used to evaluate physiological stress responses of laying hens housed in different conditions.     

Naturally occurring tuberculosis in white-tailed deer

OBJECTIVE: To determine the distribution of lesions and extent of tissues infected with Mycobacterium bovis in a captive population of white-tailed deer. DESIGN: Cross-sectional study. ANIMALS: 116 captive white-tailed deer. PROCEDURE: Deer were euthanatized, and postmortem examinations were performed. Tissues with gross lesions suggestive of tuberculosis were collected for microscopic analysis and bacteriologic culture. Tissues from the head, thorax, and abdomen of deer with no gross lesions were pooled for bacteriologic culture. Tonsillar, nasal, oral, and rectal swab specimens, fecal samples, and samples of hay and pelleted feed, soil around feeding sites, and water from 2 natural ponds were collected for bacteriologic culture. RESULTS: Mycobacterium bovis was isolated from 14 of 116 (12%) deer; however, only 9 of 14 had lesions consistent with tuberculosis. Most commonly affected tissues included the medial retropharyngeal lymph node and lung. Five of 14 tuberculous deer had no gross lesions; however, M bovis was isolated from pooled tissue specimens from the heads of each of these deer. Bacteriologic culture of tonsillar swab specimens from 2 of the infected deer yielded M bovis. Mean (+/- SEM) age of tuberculous deer was 2.5 +/- 0.3 years (range, 0.5 to 6 years). Mycobacterium bovis was not isolated from feed, soil, water, or fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Examination of hunter-killed white-tailed deer for tuberculosis commonly includes only the lymph nodes of the head. Results of such examinations may underestimate disease prevalence by as much as 57%. Such discrepancy should be considered when estimating disease prevalence.     

Effects of supplemental safflower and vitamin E during late gestation on lamb growth, serum metabolites, and thermogenesis

Twin-bearing Targhee ewes (Exp. 1, 1 yr, n = 42) and 1,182 single- and twin-bearing whiteface range ewes (Exp. 2, n = 8 experimental units over 2 yr) were used in a 2 x 2 factorial arrangement of treatments to determine the effect of supplemental energy source and level of vitamin E supplement on lamb serum metabolites and thermogenesis (Exp. 1) and on lamb growth (Exp. 2). During late gestation, ewes were individually fed (Exp. 1) or group-fed (Exp. 2) a daily supplement. Supplements were 226 g/ewe of daily safflower seed (DM basis; SS) with either 350 IU/ewe daily (VE) or no added supplemental (VC) vitamin E; or 340 g/ewe daily of a barley-based grain supplement (DM basis; GC) and either VE or VC. One hour postpartum in Exp. 1, twin-born lambs were placed in a 0 degrees C dry cold chamber for 30 min. Lamb rectal temperature was recorded every 60 s and blood samples were taken immediately before and after cold exposure. In Exp. 2, lambs were weighed at birth, at turnout from confinement to spring range (32 d of age +/- 7; turnout), and at weaning (120 d of age +/- 7). Ewes were weighed at turnout and weaning. In Exp. 1, a level of vitamin E x energy source interaction was detected (P < 0.10) for body temperature and change in NEFA and glucose concentrations. Lambs from SSVC ewes had the lowest (P = 0.01) body temperature and had decreased (P = 0.08) NEFA concentration. The SS lambs tended to have decreased (P < 0.11) concentrations of blood urea N (BUN) and thyroxine at 0 min than did lambs born to GC ewes. After 30 min of cold exposure, SS lambs had increased and GC lambs had decreased BUN, triiodothyronine, and triiodothyronine:thyroxine concentrations (P < 0.10). In Exp. 2, kilograms of lamb per ewe at turnout and weaning and lamb survival at weaning were greater (P < 0.07) for GC than SS lambs. Based on the decreased body temperature in SSVC lambs at birth, the greater change in BUN during the cold exposure for SS than GC lambs, and the decreased survival rate for SS than GC lambs, SSVC-supplemented ewes appeared to give birth to lambs with an apparently decreased energetic capacity. This may compromise the ability of the newborn lamb to adapt to extreme environmental conditions.     

Investigation of the transmission of Mycobacterium bovis from deer to cattle through indirect contact

OBJECTIVE: To investigate the infection of calves with Mycobacterium bovis through oral exposure and transmission of M. bovis from experimentally infected white-tailed deer to uninfected cattle through indirect contact. ANIMALS: 24 11-month-old, white-tailed deer and 28 6-month-old, crossbred calves. PROCEDURE: In the oral exposure experiment, doses of 4.3 x 10(6) CFUs (high dose) or 5 x 10(3) CFUs (low dose) of M. bovis were each administered orally to 4 calves; as positive controls, 2 calves received M. bovis (1.7 x 10(5) CFUs) via tonsillar instillation. Calves were euthanatized and examined 133 days after exposure. Deer-to-cattle transmission was assessed in 2 phases (involving 9 uninfected calves and 12 deer each); deer were inoculated with 4 x 10(5) CFUs (phase I) or 7 x 10(5) CFUs (phase II) of M. Bovis. Calves and deer exchanged pens (phase I; 90 days' duration) or calves received uneaten feed from deer pens (phase II; 140 days' duration) daily. At completion, animals were euthanatized and tissues were collected for bacteriologic culture and histologic examination. RESULTS: In the low- and high-dose groups, 3 of 4 calves and 1 of 4 calves developed tuberculosis, respectively. In phases I and II, 9 of 9 calves and 4 of 9 calves developed tuberculosis, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that experimentally infected deer can transmit M. bovis to cattle through sharing of feed. In areas where tuberculosis is endemic in free-ranging white-tailed deer, management practices to prevent access of wildlife to feed intended for livestock should be implemented.     

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.     

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.     

Experimental persistent infection with bovine viral diarrhea virus in white-tailed deer

Bovine viral diarrhea virus (BVDV) infections cause substantial economic losses to the cattle industries. Persistently infected (PI) cattle are the most important reservoir for BVDV. White-tailed deer (Odocoileus virginianus) are the most abundant species of wild ruminants in the United States and contact between cattle and deer is common. If the outcome of fetal infection of white-tailed deer is similar to cattle, PI white-tailed deer may pose a threat to BVDV control programs. The objective of this study was to determine if experimental infection of pregnant white-tailed deer with BVDV would result in the birth of PI offspring. Nine female and one male white-tailed deer were captured and housed at a captive deer isolation facility. After natural mating had occurred, all does were inoculated intranasally at approximately 50 days of pregnancy with 10(6) CCID(50) each of a BVDV 1 (BJ) and BVDV 2 (PA131) strain. Although no clinical signs of BVDV infection were observed or abortions detected, only one pregnancy advanced to term. On day 167 post-inoculation, one doe delivered a live fawn and a mummified fetus. The fawn was translocated to an isolation facility to be hand-raised. The fawn was determined to be PI with BVDV 2 by serial virus isolation from serum and white blood cells, immunohistochemistry on skin biopsy, and RT-PCR. This is the first report of persistent infection of white-tailed deer with BVDV. Further research is needed to assess the impact of PI white-tailed deer on BVDV control programs in cattle.     

Light and electron microscopic comparisons of cutaneous fibromas in white-tailed and mule deer

Cutaneous fibromas of white-tailed deer (Odocoileus virginianus), when compared with normal skin of the same species, had a thinner basement membrane; thickened stratum spinosum with numerous melanocytes, desmosomes, polyribosomes, and tonofilaments; focal hyperplasia of the stratum granulosum containing numerous large, electron-dense keratohyalin granules with irregular borders and containing occasional cells with diffuse intranuclear virus particles; and a moderately thickened stratum corneum with (although rarely) small crystalline arrays of virus particles. Normal mule deer (Odocoileus hemionus) skin was structurally similar to that of the white-tailed deer. Mule deer fibromas were similar to those in white-tailed deer, except for diffuse thickening of the stratum granulosum (the cells of which contained large keratohyalin granules of various electron densities with occasional composite granules) and except for a markedly thickened stratum corneum that contained numerous intranuclear viral inclusions. In negatively stained homogenates of tumors from both deer species, viral particles resembled papillomaviruses.     

MHC class II-restricted, CD4(+) T-cell proliferative responses of peripheral blood mononuclear cells from Mycobacterium bovis-infected white-tailed deer

White-tailed deer are significant wildlife reservoirs of Mycobacterium bovis for cattle, predators, and, potentially, humans. Infection of cattle with M. bovis stimulates an antigen-specific T-cell response, with both CD4(+) and CD8(+) cells implicated in protective immunity. Few studies, however, have examined lymphocyte subset responses to experimental M. bovis infection of white-tailed deer. In this study, a flow cytometric proliferation assay was used to determine the relative contribution of individual peripheral blood mononuclear cell subsets of M. bovis-infected white-tailed deer in the recall response to M. bovis antigen. Naive deer were challenged with M. bovis by cohabitation with infected deer. These M. bovis-challenged deer developed significant in vivo (delayed-type hypersensitivity) and in vitro (proliferative) responses to M. bovis purified protein derivative (PPD). At necropsy, typical tuberculous lesions containing M. bovis were detected within lungs and lung-associated lymph nodes of infected deer. The predominant subset of lymphocytes that proliferated in response to in vitro stimulation with PPD was the CD4(+) subset. Minimal proliferative responses were detected from CD8(+), gamma delta TCR(+), and B-cells. Addition of monoclonal antibodies specific for MHC II antigens, but not MHC I or CD1 antigens, abrogated the proliferative response. Together, these findings indicate that while CD4(+) cells from infected deer proliferate in the recall response to M. bovis antigens, this response is not sufficient to clear M. bovis and immunologic intervention may require stimulation of alternate subsets of lymphocytes.