Health protocol for translocation of free-ranging elk.

When considering an elk (Cervus elaphus) restoration program, wildlife managers must evaluate the positive and negative elements of translocation. We prepared this protocol to give an overview of health considerations associated with translocation of elk, with an emphasis on movement of free-ranging elk from western North America to the southeastern USA. We evaluated infectious agents and ectoparasites reported in elk from two perspectives. First, we made a qualitative estimate of the ability of the agent to be introduced and to become established. This was done using a selected set of epidemiologic factors. Second, if there was a good possibility that the organism could become established in the release area, the potential pathological consequences for elk and other wildlife, domestic animals, and humans were assessed via examination of the literature and consultation with other animal health specialists. The results of these evaluations were used to classify infectious agents and ectoparasites as low risk (n = 174), unknown risk (n = 10), and high risk (n = 9). We classified Anaplasma marginale, Anaplasma ovis, Mycobacterium paratuberculosis, Pasteurella multocida serotype 3, Elaphostrongylus cervi, Dicrocoelium dendriticum, Fascioloides magna, Echinococcus granulosus, Dermacentor albipictus, and Otobius megnini as unknown risks. High risk infectious agents and ectoparasites were the agent of chronic wasting disease, Brucella abortus, Mycobacterium bovis, Dermacentor andersoni, Ixodes pacificus, and Psoroptes sp. Parelaphostrongylus tennis, Elaeophora schneideri, and a Babesia sp. are parasites endemic in the southeastern USA that may present a "reverse risk" and adversely affect elk if released in some parts of the region. We developed a five-component protocol to reduce the risk of introduction of high risk infectious agents and ectoparasites that included: (1) evaluation of the health status of source populations, (2) quarantines, (3) physical examination and diagnostic testing, (4) restrictions on translocation of animals from certain geographic areas or populations, and (5) prophylactic treatment.

The effect of dietary aflatoxin on wild turkey poults.

Aflatoxins, toxic metabolites of Aspergillus flavus or Aspergillus parasiticus, cause poor feed utilization, decreased weight gains, depressed immune function, liver dysfunction, coagulation abnormalities, and death in a wide variety of species including humans. Conservationists have become concerned that increasingly popular wildlife feeding or baiting practices could expose wildlife to toxic amounts of aflatoxin-contaminated grains. In particular, the effects of aflatoxins on the wild turkey (Meleagris gallopova silvestris) are of concern because the conspecific domestic turkey is highly susceptible to aflatoxins. To evaluate the effect of dietary aflatoxin on wild turkeys, four groups of 4-mo-old wild turkeys were fed diets containing either 0, 100, 200, or 400 micrograms aflatoxin/kg feed for 2 wk in September and October 1996. Aflatoxin-fed poults had decreased feed consumption and weight gains as compared with control poults. Decreased liver-to-body weight ratios, liver enzyme alterations, slightly altered blood coagulation patterns, and mild histologic changes indicated low-level liver damage. Compromise of cell-mediated immunity was indicated by decreased lymphoblast transformation. The effects were apparent in all treatment groups to variable levels, but significant differences most often were found at 400 micrograms aflatoxin/kg feed. This study shows that short-term aflatoxin ingestion by wild turkeys can induce undesirable physiologic changes; therefore, exposure of wild turkeys to feeds containing aflatoxin levels of 100 micrograms aflatoxin/kg feed or more should be avoided.

Responses of red foxes to first and second infection with Sarcoptes scabiei.

The clinical response of red foxes (Vulpes vulpes) to the mange mite, Sarcoptes scabiei, was characterized by infection of five, 4-mo-old red foxes with S. scabiei originally isolated from a wild red fox. The infected foxes and three uninfected control foxes were monitored with weekly complete blood counts and biweekly serum chemistry profiles, hypersensitivity tests, and evaluation of skin biopsies. After 7 wk, the foxes were treated and held free of infection for 2 mo. Six foxes, three previously infected and three with no history of exposure, were then infected with the same isolate of S. scabiei and followed for another 7 wk; two additional previously infected foxes were held as treatment controls, and two foxes with no history of exposure as naive controls. All infected foxes developed significant immediate (Type I) hypersensitivity reactions to a S. scabiei mite extract within 2 wk of exposure and maintained this reaction as long as 4 mo after clearance of mites. Pronounced mast cell hyperplasia and infiltration with eosinophils were the earliest inflammatory cell responses noted in biopsy samples from infected foxes and were maintained throughout infection. Infected foxes also showed significant increases in white blood cell counts, due primarily to increases in numbers of circulating neutrophils and eosinophils. Clinical response, severity of disease, and relative numbers of mites per cm2 of skin of previously infected foxes and foxes undergoing their first infection did not differ. These results show that red foxes develop strong immediate hypersensitivity reactions to S. scabiei but, under our experimental conditions, did not exhibit resistance to reinfection.

Diseases diagnosed in red foxes from the southeastern United States.

Diagnostic findings on 51 red foxes (Vulpes vulpes) from the southeastern United States examined from 1967 to 1995 were reviewed. Etiologic diagnoses included sarcoptic manage (n = 33), traumatic injury and associated complications (n = 5), suspected canine distemper (n = 2), capture myopathy (n = 1), congenital absence of guard hairs (n = 1), intradermal tick infestation (n = 1), otodectic manage (n = 1), and toxicosis (n = 1). The cause of morbidity was not determined for three of the foxes, and three others were classified as normal animals. Sarcoptic manage was diagnosed in 65% of the red foxes, was found in foxes submitted from four of the eight southeastern states represented, and was seen in 19 of 29 yr covered by this study.

Host defense responses associated with experimental hemorrhagic disease in white-tailed deer.

Our objectives were to examine the immunity conferred by epizootic hemorrhagic disease virus serotype 2 (EHDV-2) infection in white-tailed deer (Odocoileus virginianus) and determine if this immunity was protective during challenge with homologous (EHDV-2) or heterologous (bluetongue virus serotype 10; BTV-10) virus. Trials were conducted in the fall of 1992 and 1993. In the first experiment, naive white-tailed deer were infected intradermally and subcutaneously with EHDV-2 and monitored via physical examinations, complete blood counts, alpha and beta interferon (IFN) assays, viral isolation, and serology. Infected deer had a wide range of clinical signs in response to infection. Eleven of the 16 deer had body temperature elevations > or = 0.5 C between post-infection day (PID) 4 and 8. Infected deer had decreased lymphocyte counts between PID 6 and 10 that returned to normal levels by PID 17. Severely lymphopenic animals had the most severe clinical signs; five of 10 deer with lymphocyte counts less than 1000 cells/microliters succumbed to the infection. Viremia was detected in all 16 EHDV-2 infected animals by PID 4, and peak viremias occurred between PID 4 and PID 10. Three deer remained viremic until PID 56, the study endpoint. Interferon was first detected between PID 2 and 6. Peak alpha and beta IFN levels coincided with peak viremia in 11 deer. Precipitating and neutralizing antibodies were detected in infected deer by PID 10. In the second experiment, convalescent deer were challenged subcutaneously and intradermally with either EHDV-2 or BTV-10 and similarly monitored. Virus was detected in the blood of all four deer challenged with BTV-10, but viremia was not detected in three EHDV-2-challenged deer. Temperature fluctuations, blood cell parameter changes, and IFN and antibody responses seen in BTV-10-challenged deer were similar to those seen in the initial experiment. Deer challenged with EHDV-2 had mildly increased temperatures, but minimal IFN response and lymphocyte alterations.

Shell disease in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) in a Georgia (USA) lake.

A disfiguring shell disease was detected in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) from Lake Blackshear, Georgia (USA). The turtles used were part of a mark-recapture study conducted from September 1991 to June 1993. Histologic changes on four turtles included acute segmental necrosis of the epidermis, followed by ulceration, necrosis of the underlying dermis and dermal bone, and exaggerated remodeling of bone. Additional findings included visceral inflammatory lesions and bacterial infection, sepsis and marked trematode ova granulomatosis. The cause of the shell lesions was not determined.

Evaluation of low-level aflatoxin in the diet of white-tailed deer.

We evaluated the response of white-tailed deer (WTD) (Odocoileus virginianus) to dietary aflatoxin. Fourteen 4-to-5-mo-old WTD were used in this 8-wk study, conducted between November 1993 and January 1994. Seven animals received a ration containing 800 parts per billion (ppb) total aflatoxin (AF). Seven control animals received the same ration without AF. At 0, 1, 3, 6 and 8 wk, feed consumption, feed conversion, liver enzymes, bile acid levels, and immune function via lymphocyte proliferation assays and delayed type hypersensitivity reactions were determined. At the conclusion of the 8-wk feeding trial, deer were euthanized and necropsied. Clinical illness was not evident in any of the animals, but by the end of the study, AF-fed deer had reduced feed consumption and body weight as compared to control deer; the differences were not statistically significant. The AF-exposed group had a significant increase (P = 0.03) in serum bile acid concentration as compared to control deer. Two AF-exposed deer had gross and histologic hepatic lesions indicative of a mild degenerative hepatopathy. Residues of an aflatoxin metabolite, aflatoxin M1, were found in the livers of all treated animals. No differences in immune function were detected between the two groups. We conclude that consumption of 800 ppb AF in the diet of young WTD over an 8-wk period can produce subclinical hepatic injury.

Potential consequences and problems with wildlife contraceptives.

This presentation reviews multiple wildlife health considerations associated with contraception of wildlife, at the level of both the individual animal and wildlife population. Review of the literature demonstrates that many contraceptives studied in the past have had potential adverse effects characterized as follows: harmful effects on pregnant animals, inhibition of parturition or dystocia, changes in ovarian structure or function, changes in sex ratio, changes in lactation or mammary glands, impact on fertility of young, changes in testicular structure or function, changes in secondary sex characteristics, changes in bodyweight, changes in behaviour, changes in annual breeding season, other physiologic and pathologic changes, abscesses or inflammatory reactions, toxicity, interference with diagnostic tests and ecological alterations. Concern is expressed that the use of immunocontraception could create genetic changes in the target population that would influence disease resistance. The use of infectious agents as vectors to deliver immunocontraceptives was not considered wise animal health management because the product will be a new reproductive disease that could be difficult to contain within the target population. Criteria that need to be fulfilled for the safe use of contraceptives in wildlife are offered.

Hemorrhagic disease in white-tailed deer in Texas: a case for enzootic stability.

Although antibodies to viruses in both the epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) sero-groups have been reported from white-tailed deer (Odocoileus virginianus) in Texas (USA), there are few reports of hemorrhagic disease (HD) in these populations. To understand the extent and diversity of exposure to the North American EHDV and BTV serotypes in these deer populations, we serologically tested 685 white-tailed deer collected from November 1991 through March 1992 throughout their range in Texas. Overall, 574 (84%) of deer had antibodies to EHDV or BTV. Prevalence estimates varied according to ecological region, from 57% in the Gulf Prairies to 100% in the northwest Edwards Plateau. Based on serum neutralization tests, the deer had evidence of previous exposures to multiple EHDV and BTV serotypes, with evidence of exposure to two to five serotypes detected in each ecological region. The apparent lack of HD in relation to this high antibody prevalence cannot be explained, but may be related to enzootic stability in which a near perfect host-virus relationship exists.

Potential impact of wildlife on the tropical bont tick eradication program in the Caribbean.

Wildlife are hosts for A. variegatum throughout its range in Africa, and have been demonstrated to serve as hosts for larvae and nymphs on several islands in the Caribbean. Studies conducted in the Caribbean have indicated that most larvae and nymphs and all adults of the tick feed on livestock; therefore, eradication can be approached through the treatment of livestock. However, since small numbers of animals such as the mongoose are infested with larvae and nymphs, these animals may represent a short-term alternative host system. Such infestations potentially could result in a presence of small numbers of ticks for up to 46 months after the onset of a treatment program and could serve as a source to reinfest livestock during the eradication process. Wildlife surveys have been conducted in St. Croix, Puerto Rico, Guadeloupe, and Antigua; but the host status of species present on other islands such as the white-tailed deer on St. Kitts has not been evaluated. As part of the eradication program, the species of wildlife present on each island and the association of these species with infested pastures should be determined. Surveys of wildlife can be used during the eradication to determine if wildlife are serving as an alternative host system. Cattle egrets may serve to disseminate the tick among islands in the region, and to areas outside the region. Cattle egrets have been shown to serve as hosts for the tick, and to move throughout the eastern Caribbean and to North America. The potential for interisland dissemination of ticks by cattle egrets, as well as by other means, substantiates the need for a region-wide eradication program.

Experimental infection of Culicoides lahillei (Diptera: Ceratopogonidae) with epizootic hemorrhagic disease virus serotype 2 (Orbivirus: Reoviridae).

To evaluate the vector competence of Culicoides lahillei Lutz for epizootic hemorrhagic disease (EHD) viruses, wild-caught females were allowed to feed on 4 viremic white-tailed deer, Odocoileus virginianus Zimmermann, experimentally infected with EHD virus serotype 2 (EHDV-2). Colonized C. variipennis sonorensis (Coquillett) were included as a positive control. Virus was not isolated from 13 C. lahillei tested 4-15 d after feeding on deer with viremias of 3.0 or <2.1 log10 tissue culture infectious dose50 (TCID50) virus per milliliter of blood. Virus was isolated from 6 of 69 (8.7%) C. lahillei tested 4-15 d after feeding on deer with viremias of 5.3 or 6.0 TCID50 virus per milliliter of blood. The average amount of EHDV-2 ingested in the blood meal ranged from

Incompetence of white-tailed deer as amplifying hosts of vesicular stomatitis virus for Lutzomyia shannoni (Diptera: Psychodidae).

Sand flies, Lutzomyia shannoni Dyar, were allowed to feed on 3 white-tailed deer, Odocoileus virginiana Zimmermann, that previously had been infected with the New Jersey serotype of vesicular stomatitis (VSNJ) virus. Flies fed in the lower abdominal area of each deer on days 1-5 postinfection. A blood sample, nasal swab, and throat swab were taken during each feeding trial and examined for virus. Blood-fed flies were held for 4-5 d following the bloodmeal and tested for VSNJ virus infection. VSNJ virus was never detected in blood or from swabs taken from infected deer nor from any of the sand flies that fed on deer. The findings suggest that white-tailed deer do not fulfill the traditional concept of amplifying hosts of VSNJ virus.

Incompetence of domestic pigs as amplifying hosts of vesicular stomatitis virus for Lutzomyia shannoni (Diptera: Psychodidae).

Seven domestic pigs, Sus scrofa L., were infected by intradermal inoculation at 3 different sites with the New Jersey serotype of vesicular stomatitis (VSNJ) virus. Laboratory-reared Lutzomyia shannoni Dyar sand flies, a suspected biological vector of VSNJ virus, were allowed to feed on pigs at the lower abdomen or at sites of their own selection on days 1-7 and on day 10 postinfection. Blood samples were taken from infected swine concomitant with most feeding trials and tested for the presence of virus. Sand flies were held for up to 5 d following ingestion of blood and tested for VSNJ virus infection. Virus was not recovered from the blood of infected pigs or from any of the flies that fed on these pigs. The findings suggest that domestic pigs do not fulfill the traditional concept of amplifying hosts of VSNJ virus.

Epizootic hemorrhagic disease virus and bluetongue virus serotype distribution in white-tailed deer in Georgia.

Serum samples collected from 1,396 white-tailed deer (Odocoileus virginianus) in five areas of Georgia (USA) from 1989 to 1991 were tested for precipitating and serum neutralizing (SN) antibodies to the enzootic North American epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) serotypes. Precipitating antibodies to the EHDV or BTV serogroups, as detected by agar gel immunodiffusion (AGID) tests, were present in 35%, 29%, and 39% of deer sampled in 1989, 1990, and 1991, respectively. Significant differences (P < 0.05) in precipitating antibody prevalence were detected between physiographic regions during all years. Antibody prevalence consistently was highest in deer sampled from the Coastal Plain (77%), followed by the Piedmont (33%), Ridge and Valley (29%), Barrier Island (5%), and Blue Ridge (2%) regions. All AGID-positive samples were tested by SN tests for antibodies against all North American EHDV and BTV serotypes (EHDV serotypes 1 and 2, BTV serotypes 2, 10, 11, 13, and 17). Criteria for previous exposure to a specific serotype were either detection of monospecific results or clusters of positive results against that serotype. Serologic evidence of previous exposure to EHDV serotypes 1 and 2, and BTV serotypes 11 and 13 was detected during all years. Predominant serotypes varied among years. In general, evidence of exposure to EHDV serotype 2 appeared annually while exposure to BTV serotype 13 and EHDV serotype 1 decreased and increased, respectively. To determine serotype diversity prior to 1989, 134 AGID-positive white-tailed deer serum samples collected from 1967 to 1988 also were tested by SN. Evidence of exposure to EHDV serotypes 1 and 2 and BTV serotypes 11, 13, and 17 was detected.

Disinfection and wildlife.

Capture, handling or transport of wildlife for purposes such as research, disease monitoring, wildlife damage control, relocation, and collection of zoological specimens can create risks of disease spread. Cleaning and disinfection procedures for equipment used in these activities must be routine and designed to eliminate the spread of pathogens to either animals or humans. General methods and materials for cleaning and disinfection apply to wildlife studies. Concepts involved in preparing a protocol specific to a wildlife investigation are discussed. The control of the spread of livestock and poultry pathogens via free-ranging mammals and birds prior to disinfection of contaminated premises is approached through an accurate assessment of the problem and, where necessary, the selection of appropriate wildlife control measures. The authors discuss the development of a problem assessment, and review potential methods for use in the control of wildlife. For an accurate problem assessment, information is needed on the presence of wild mammals and birds at the site, exposure of wild mammals and birds to the pathogen, and the potential for further transmission. When wildlife control is deemed necessary, techniques may be selected to disperse or exclude animals from premises or to depopulate the site. Dispersal or exclusion from premises is appropriate when movement of animals within or away from the contaminated premises would not result in further transmission of the pathogen. Depopulation is necessary when the continued presence or dispersal of wild mammals or birds would potentially result in further spread of the disease.

Susceptibility of white-tailed deer (Odocoileus virginianus) to infection with Ehrlichia chaffeensis, the etiologic agent of human ehrlichiosis.

Although more than 320 cases of human ehrlichiosis have been diagnosed in 27 states since 1986, the reservoir host or hosts remain unknown. Since antibodies reactive to Ehrlichia chaffeensis, the etiologic agent of human ehrlichiosis, have been found in white-tailed deer (Odocoileus virginianus), we experimentally evaluated the susceptibilities of four white-tailed deer to infection with E. chaffeensis and Ehrlichia canis, a closely related species. A fifth deer served as a negative control. Isolation and nested PCR amplification results from peripheral blood indicated that E. chaffeensis circulated for at least 2 weeks. The deer developed antibodies to E. chaffeensis by day 10 after inoculation, but there was no indication of clinical disease. Immunohistochemical staining identified E. chaffeensis within macrophage-type cells in lymph nodes. The deer inoculated with E. canis did not become infected and did not seroconvert. These results indicate that white-tailed deer can support an E. chaffeensis infection with resulting rickettsemia of at least 2 weeks. The resistance to infection and the absence of seroconversion upon exposure to E. canis indicate that antibody responses previously detected among wild deer are not E. canis cross-reactions. The role of deer as competent reservoirs in the life cycle of E. chaffeensis remains to be explored with suspected tick vectors.

Field evaluation of baits and baiting strategies for delivering oral vaccine to mongooses in Antigua, West Indies.

A field study was conducted on Antigua, West Indies, to determine the feasibility of delivering an oral rabies vaccine or population control agent to free-ranging mongooses (Herpestes javanicus). Two biomarkers (tetracycline hydrochloride [THCL] and DuPont Oil Blue A dye) and two bait types (DuPont polymer fish meal and polyurethane foam) were used to bait three study sites. Four hundred polymer baits containing both biomarkers were distributed at 36 central point bait stations (11 baits/station) on an 80 ha study site (5 baits/ha); 69% of the mongoose population consumed one or more baits. Two thousand baits containing THCL and 400 baits containing DuPont dye were distributed on two additional 100 ha study sites (24 baits/ha). Polymer fish meal baits were used on the first site and polyurethane baits on the second site. Based on the presence of biomarkers in bone or soft tissue, 96 to 97% of the mongooses at both sites consumed at least one bait. We conclude that oral baiting of mongooses is a feasible method for delivery of vaccines for the control of rabies in this species.

Potential role of cattle egrets, Bubulcus ibis (Ciconiformes: Ardeidae), in the dissemination of Amblyomma variegatum (Acari: Ixodidae) in the eastern Caribbean.

The potential role of cattle egrets, Bubulcus ibis (L.), in the dissemination of Amblyomma variegatum (F.) in the eastern Caribbean was evaluated. The status of cattle egrets as hosts for A. variegatum in Antigua was determined during seven survey periods between September 1988 and May 1991. The mean prevalences of infestation by larvae and nymphs were 2.3 and 0.5%, respectively. The mean intensity and relative density of infestation by larvae were 8.8 (SD = 9.80) and 0.2 (SD = 0.28), respectively. Cattle egrets examined in Guadeloupe during February-March 1991 were not infested but 5.9% were infested by larvae during June-July 1991. Interisland movement of cattle egrets was evaluated relative to emigration of birds captured and marked in Antigua and Guadeloupe. During this aspect of the study, 1,129 cattle egrets were captured, marked, and released. Of 195 sighting reports received, 56 were determined to be independent sightings. Emigration of cattle egrets included movement of birds marked in Antigua or Guadeloupe to 14 Caribbean islands and the Florida Keys. Interisland movement occurred in each of the discrete observation periods during the 3-yr study period. The rate of emigration per period ranged from 1.2 to 12.9%. That cattle egrets served as hosts for immature A. variegatum in the eastern Caribbean and moved between islands in the region demonstrates that these birds could serve as disseminators of the tick. Estimates of the numbers of infested cattle egrets emigrating from Antigua and Guadeloupe ranged from 0 to 0.24% of the current populations.

Feral swine as a potential amplifying host for vesicular stomatitis virus New Jersey serotype on Ossabaw Island, Georgia.

Sentinel feral swine (Sus scrofa) on Ossabaw Island, Georgia (USA), were serologically monitored for antibodies to vesicular stomatitis New Jersey serotype (VSNJ) virus from 17 April to 27 August 1990. Seroconversions to VSNJ virus were detected in 24% of swine island-wide. Differences in the incidence of seroconversion were detected between swine sampled in the Pleistocene and Holocene formations of the island suggesting that the presence of virus is forest type dependent. Based on the consistency in onset and spatial distribution of seroconversions with data from 1981 to 1985, this is a very stable host-parasite system. Sequential virus isolation attempts from nasal swabs, tonsil swabs, and blood were made on a subsample of 54 sentinel swine from 9 May to 4 July 1990. The VSNJ virus was isolated from five swine from 16 May to 20 June. Vesicular lesions were detected on only two of these animals. Although infections in these feral swine were short-lived (< 7 days) and were followed by a strong neutralizing antibody response, VSNJ virus was detected in a single group of swine for a period exceeding 1 month. From these data, it appears that feral swine could provide a source of virus to feeding arthropods for extended periods of time. The failure to detect a viremia in these animals, however, indicates that a source other than blood may be required for transmission to occur.