Sixteen-bird groups (sexes equal) of adult mallards (Anas platyrhynchos) were orally dosed with eight #4 steel shot, eight #4 lead shot, eight BB-size tungsten-iron shot, eight BB-size tungsten-polymer shot, or were sham-dosed and maintained for 30 days (16 January 1996 to 15 February 1996). Half of the lead-dosed ducks (five males, three females) died during the study, whereas no ducks died in the other dosage groups. For lead-dosed ducks, hematocrit and hemoglobin concentration were decreased on day 15 of the trial, but not on day 30. Delta aminolevulinic acid dehydratase activity in lead-dosed ducks was lower when compared to steel-dosed ducks only. Plasma activities of selected enzymes were elevated in lead-dosed ducks when compared to enzyme activities of ducks in the other groups. For lead-dosed ducks, relative heart, liver, and kidney weights increased in comparison to relative weights of those organs of ducks in other groups. Histology of tissues indicated that renal nephrosis accompanied by biliary stasis was present in the eight lead-dosed ducks that died. For the eight lead-dosed ducks that survived, six had mild to severe biliary stasis. Mild biliary stasis was noted in five tungsten-iron dosed ducks and three tungsten-polymer dosed ducks. Amounts of lead in the femur, liver, and kidneys were higher in lead-dosed ducks than in ducks of the other four groups. Small amounts of tungsten were detected in the femur and kidneys of two tungsten-polymer dosed ducks. Higher concentrations of tungsten were detected in the femur, liver, and kidneys of all tungsten-iron dosed ducks. The rate of shot erosion was highest (80%) for the tungsten-polymer shot, followed by tungsten-iron (55%), lead (50%), and steel shot (33%). Results indicated that tungsten-iron or tungsten-polymer shot (8 shot/duck) orally administered to mallards did not adversely affect them during a 30-day trial.

Effects of five lead (Pb), iron (Fe), or bismuth (Bi)/tin (Sn) alloy shot embedded in the breast muscles of game-farm mallards (Anas platyrhynchos) were studied from 28 March 1994 through 27 March 1995. We detected no differences in the mean survival times, mean hematocrits, or mean body weights among the three shot types. Connective tissue encapsulated Pb and Bi/Sn shot but only slight changes occurred in tissues surrounding the shot. Recovered Pb and Bi/Sn shot were essentially unchanged in appearance and weight. A thin zone of “oxide” surrounded Fe shot with a slight inflammatory response and a small amount of scarring adjacent to the embedded shot. Fe shot decreased slightly in weight while embedded. Bacterial infections were absent in all dosed ducks. Mean weights of kidneys, livers, and gonads did not vary by type of shot. Kidneys and livers of Bi-dosed ducks had higher concentrations of Bi than in Pb- and Fe-dosed ducks. Muscle and blood showed no differences in Bi concentrations among doses. We found no histological dose-related effects in kidneys, liver, and gonads from the embedded shot.

Captive white-winged doves (Zenaida asiatica) were exposed to various levels of methyl parathion (MP) in drinking water to determine effects on brain and blood cholinesterase (ChE) activity. We conducted two experiments to test the influence of MP dose (the amount of MP actually ingested), MP concentration (the amount of MP per unit water), and exposure duration (number of days exposed to a constant MP concentration) on ChE activity. Plasma ChE activity was not useful in predicting brain ChE activity. Methyl parathion concentration had a greater influence on plasma and brain ChE activity levels than dose or time of exposure. These results contribute to the evaluation of irrigation water as a route of exposure of wildlife to pesticides.

The connection between bald eagles (Haliaeetus leucocephalus) and the consumption of waterfowl, lead shotshell pellet ingestion, and subsequent lead exposure is well documented in the United States and is presumed also to be widespread in Canada. We compared blood lead concentrations in samples from bald eagles ranging in age from 0.5- to 1.5-yr-old at Galloway Bay, Saskatchewan, Canada (n = 97) during October–November, 1992–95 and Hauser Lake, Montana, USA (n = 81) during October-December, 1990–94, within the Great Plains region of North America. Abundant prey are available in the form of hunter-injured waterfowl in Saskatchewan and spawning salmon in Montana; both areas attract large numbers of mid-continent bald eagles during fall migration. Blood lead concentrations suggestive of recent lead exposure (>0.201 μg/ml) were found in 32% and 8% of eagles at Hauser Lake and Galloway Bay, respectively, when samples from each study area were analyzed independently at two laboratories. To determine if this difference was an artifact of interlaboratory variation, we determined a correction factor by reanalyzing 14 Saskatchewan blood samples at each laboratory and predicted blood lead concentrations from Hauser Lake had the samples been analyzed at the Canadian laboratory. Adjusted blood lead concentrations of samples from Hauser Lake indicated that 21% of eagles were recently exposed to lead, a proportion not significantly different from the proportion of the same exposure category at Galloway Bay. Our data do not support the supposition that a large proportion of bald eagles feeding on waterfowl in areas of high hunting pressure will be exposed to lead via consumption of lead shotshell pellets in waterfowl.

Florida sandhill cranes (Grus canadensis pratensis) were conditioned to confinement in an enclosure for 7 days, 6 hr a day. On day 8, cranes were catheterized and then confined in an enclosure. Venous blood (2 ml) was collected through the catheter and an attached IV line immediately before (−60 min) and 60 min after (0 min) confinement. Using a randomization table and a restricted cross-over experimental design, cranes were injected intravenously with either saline (control) or adrenocorticotropic hormone (ACTH; cosyntropin, Cortrosyn®; 0.25 mg). At 30, 60, 120, 180, 240 and 300 min after injection, blood samples were collected and assayed for corticosterone. The cranes receiving ACTH increased their serum corticosterone concentrations as much as fivefold above baseline concentrations. Serum corticosterone concentrations remained significantly elevated for approximately 60 min after ACTH stimulation. Physical restraint and catheterization caused an increase in serum corticosterone almost comparable to that induced by ACTH stimulation. In cranes injected with saline, serum corticosterone decreased within 1 hr after physical restraint and catheterization, and remained at lower levels throughout the remaining 5 hr of confinement.

A model was derived for disease transmission in dynamic host populations and its application was demonstrated in forecasting possible outcomes of a bovine tuberculosis (Mycobacterium bovis) epidemic in a white-tailed deer (Odocoileus virginianus) population. The approach was mechanistic, based disease transmission on the probability of each susceptible individual becoming infected per unit time, and afforded the flexibility necessary to model epidemics in dynamic wildlife populations. This approach was applied to a sex- and age-structured deer population model. This model predicted that tuberculosis prevalence in a white-tailed deer population could rise from approximately 3% to about 21% over 25 yr, and that neither lowered deer survival nor lowered transmission would be completely effective in eliminating disease from the population. Maternal transmission appeared unimportant to modeled tuberculosis dynamics; in contrast, disease was not maintained for >15 yr in models lacking lateral transmission.

Blood samples from six mule deer (Odocoileus hemionus hemionus), 15 black-tailed deer (O. hemionus columbianus), and 29 elk (Cervus elaphus nannodes) were assayed for human monocytic and human granulocytic ehrlichiosis (HGE) by polymerase chain reaction (PCR), DNA sequencing, and serology to determine whether or not cervids are involved in the maintenance of these potential human pathogens in California (USA). The deer were sampled in August to October 1992–95. The 29 tule elk from Point Reyes National Seashore were sampled in August 1997. All deer were seronegative for antibodies to HGE/Ehrlichia equi, while the E. equi seroprevalence among elk was 17%. The 16S rDNA PCR prevalence in deer was 38% (in mule deer and black-tailed deer) for Ehrlichia-like sp. of white-tailed deer, 5% (one black-tailed deer only) for E. equi, and 0% for E. chaffeensis. The PCR prevalence in elk was 0% for Ehrlichia-like sp. of white-tailed deer, 31% for E. equi, and 0% for E. chaffeensis. The E. equi from two positive elk samples was successfully propagated in HL-60 cell cultures. DNA sequencing confirmed that the Ehrlichia-like sp. sequences from deer in California were closely related to sequences reported from white-tailed deer from Oklahoma and Georgia. The E. equi strain from deer and elk resembled other E. equi strains from California. These results suggest that cervids may be important in the natural maintenance of E. equi in California.

The sera of 195 hunter-killed feral pigs (Sus scrofa), collected in New South Wales (Australia) from April to November 1995, were screened against a reference panel of 14 Leptospira interrogans serovars using a microscopic agglutination test (MAT). The panel represented those serovars previously isolated from wild and domestic mammals in mainland Australia. Anti-leptospiral agglutinins were detected in 20% of the sera tested and included nine L. interrogans serovars. The majority of serological reactors (63%) were to L. interrogans serovar pomona. Sera from 26% of immunoreactors cross reacted with antigens from one or more serovars. No differences were noted in the prevalence of L. interrogans antibodies between the sexes, or between pigs from areas of low and high rainfall. The implications of leptospirosis in feral pigs on the transmission of leptospires to wildlife, livestock, and humans are discussed.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of type C botulinum toxin (Clostridium botulinum) in wild birds. This simple, antigen-capture ELISA utilizes polystyrene immunosticks as the solid substrate, chicken antitoxin (IgY) as the coating antibody, rabbit antitoxin as the primary antibody, and peroxidase-labeled goat-anti-rabbit as the secondary antibody. To evaluate the immunostick ELISA as a diagnostic test for avian botulism, known concentrations of toxin were added to heparinized blood collected from healthy birds and tested by both the ELISA and mouse bioassay. Also, blood samples from 236 bird carcasses submitted to the National Wildlife Health Center (NWHC) for cause of death determinations were tested by both procedures. Using ≤ 0.5 ml as the test volume for both procedures, the ELISA was less sensitive, detecting 0.25 ng/ml of toxin compared to 0.12 ng/ml for the mouse bioassay. Using the same volume of test sample for diagnostic submissions (≤ 0.5 ml), the ELISA was positive for 60% of the 149 clinically-diagnosed cases of botulism, whereas the mouse bioassay was positive for 79%. However, we demonstrated that with larger sample volumes (≥ 1.0 ml), the sensitivity of the ELISA may be equivalent or better than the mouse test due to the concentrating effect of the ELISA procedure. These preliminary results suggest that when adequate sample volumes are available, the immunostick ELISA can replace the mouse test for the diagnosis of botulism in wild birds.

A field trial to evaluate the efficacy of an oral vaccinia-rabies glycoprotein recombinant virus vaccine in controlling epidemic raccoon (Procyon lotor) rabies was conducted by distributing 180,816 doses (10^8.2TCID₅₀/ml) of vaccine in wax ampules within fish-meal polymer baits at a rate of 64 doses/km²/treatment throughout a 552 km² area, forming an 18 km wide band across the northern Cape May Peninsula of New Jersey (USA). Vaccination treatments were conducted in the spring and fall between May 1992 and October 1994 from a helicopter along ecotones and from motor vehicles along roads. Vaccine-laden baits were removed by animals from tracking stations within 3 wk and 61% of the identifiable tracks were those of raccoons. Tetracycline incorporated in the baits as a biomarker was detected in 155 (73%) of the vaccination area raccoons following the fall 1993 and spring 1994 vaccinations. Eleven (61%) of the raccoons sampled in the same time period seroconverted (≥0.5 IU) in response to rabies virus glycoprotein. A raccoon diagnosed with rabies from the northern border of the vaccination area on 30 April 1993 provided the first evidence that the barrier was being challenged by the rabies epidemic. The prevalence of rabies in raccoons from the vaccination area for the first year (10%, n = 96) and second year (8%, n = 61) of challenge was reduced more than six-fold by vaccination compared to unvaccinated raccoons from northern adjacent surveillance areas during the corresponding first (65%, n = 189) and second years (53%, n = 43). Vaccination also effectively reduced by three-fold the rate at which the epidemic moved through the raccoon population (15 km/yr). The breach of the vaccination area resulted in a resumption of the high rate (43 km/yr) of epidemic movement and a significant nine-fold increase in rabies prevalence (77%, n = 47). The maximum linear movement (12.9 km) among five ear-tagged rabid raccoons in the study area was significantly greater than that of 19 normal radio-collared raccoons (2.58 km) in the area. These large movements of rabid raccoons, together with relocation of nuisance raccoons, spillover of raccoon rabies in skunks (Mephitis mephitis) and other species, insufficient funding and a decision to discontinue the program in 1994 (which could have resulted in insufficient population immunity among raccoons in the vaccination area) may have contributed to the eventual breach of the barrier.

Rabies is a widespread zoonosis that recently reached epidemic proportions in gray foxes (Urocyon cinereoargenteus) in central Texas. The objectives of this study were to determine bait and attractant preferences among captive gray foxes, to determine the behavioral responses of gray foxes to selected bait-attractant combinations, and to evaluate baits as a delivery mechanism of oral rabies vaccines. Trials were conducted to determine bait preferences of captive gray foxes to selected baits and attractants. Tested baits consisted of a polymer-bound cube made of either dog food meal or fish meal, a polymer-bound cylinder made of dog food meal, and a wax-lard cake that was enhanced with marshmallow or chicken flavoring. Attractants were additives to baits that exuded sweet, sulfurous, fruity, fatty, cheesy, honey, and fishy odors and flavors. Captive gray foxes (n = 31) exhibited a preference for marshmallow wax cakes and polymer dog food baits with a lard interior and granulated sugar exterior. However, gray foxes exhibited chewing behaviors consistent with ingesting an oral vaccine only with the wax cake baits.

From August 1995 to August 1997, six of 18 common dolphins (Delphinus delphis) that stranded along beaches of southern California (USA) tested antibody positive for dolphin morbillivirus (DMV). Titers ascertained by virus neutralization ranged from 1:50 to 1:910 while those determined by ELISA ranged from 1:80 to 1:195. The first individual to strand survived and was released back into the Pacific Ocean 14 mo later. Histopathologic examination of tissues from the other five dolphins did not reveal lesions characteristic of morbilliviral disease; however, morbilliviral RNA was detected in three of the five by reverse transcriptase-polymerase chain reaction testing. This is the first report of morbilliviral infection in any marine mammal species in the northern hemisphere of the Pacific Ocean. These data indicate that DMV, or a closely related morbillivirus, is present in the Pacific Ocean and infection of common dolphins may not be associated with morbillivirus disease.

A total of 486 serum samples collected from several species of both free-living and captive cervids in Germany was examined for antibodies against malignant catarrhal fever (MCF)-associated viruses (MCFV) by a competitive-inhibition enzyme-linked immunosorbent assay (CI-ELISA). Eleven (2%) of these samples were positive for antibodies against MCFV. Among 157 serum samples collected from 16 different species of captive deer including four (7%) of 54 fallow deer and one (7%) of 14 sika deer (Cervus nippon) were seropositive. Among 329 samples from three species of free-ranging deer, including 253 roe deer (Capreolus capreolus), 22 red deer (Cervus elaphus) and 54 fallow deer (Cervus dama), only fallow deer were antibody-positive. Of the 25 fallow deer samples collected between 1990 and 1993, four (16%) were seropositive. Among 29 free-ranging fallow deer samples collected in the hunting period 1996–1997, antibodies to MCFV were detected in two (7%) of these sera. All of these fallow deer samples were collected from a circumscribed area in northern Germany. In the same area a high seroprevalence (72%) to MCFV was observed in domestic sheep (n = 50). Among 20 sheep samples (buffy coat) and 15 fallow deer samples (spleen or lymph nodes) examined for ovine herpesvirus 2 (OvHV-2) by PCR, all 20 sheep samples examined were OvHV-2 positive, but all of the 15 fallow deer samples, including seven seropositive deer, were OvHV-2 negative.

Reticuloendotheliosis in captive greater (Tympanuchus cupido pinnatus) and Attwater's (T. cupido attwateri) prairie chickens is reported for the first time. Between September 1993 and August 1994, two adult female wild-caught greater prairie chickens housed at Texas A&M University (College Station, Texas, USA) were observed with multiple subcutaneous nodules. Both birds were euthanatized. Complete necropsy examinations revealed lesions limited to the skin of each bird. Histopathologic examination of lesions revealed pleomorphic lymphoreticular cells suggestive of reticuloendotheliosis and reticuloendotheliosis virus (REV) was demonstrated in tumor tissue by polymerase chain reaction and virus isolation. Between September 1994 and June 1995, five additional greater prairie chickens and two Attwater's prairie chickens were euthanatized or found dead with evidence of lymphoreticular neoplasia in multiple organ systems. Initial testing of the captive flock in December 1994 for evidence of viremia and antibody to reticuloendotheliosis virus revealed over 50% of the tested birds were viremic, but none developed antibodies. Subsequent testing between January 1995 and January 1996 indicated that once infected with reticuloendotheliosis virus, Attwater's prairie chickens tended to remain outwardly healthy despite persistent viremia compared to infected greater prairie chickens which had higher morbidity and mortality rates within 60 to 90 days after initial detection of viremia and did not usually develop persistent viremia. Antibodies to REV were detected in only three captive greater prairie chickens and only in 1995. Six of the nine birds that were euthanatized or found dead due to reticuloendotheliosis developed viremia prior to death; three birds were not tested prior to death. Testing of free-ranging greater and Attwater's prairie chickens for reticuloendotheliosis is recommended prior to translocation or release.

During research on polar bears (Ursus maritimus) at Svalbard in April 1996, we captured two yearlings with a normal vaginal opening and a 20 mm penis containing a baculum. The penis was located caudal to the location in a normal male and was concealed within the vaginal opening by a single pair of labia. The urethral opening was situated laterally about 5 mm from the distal end of the penis. Neither of the yearlings showed signs of a Y chromosome, so both bears were regarded as female pseudohermaphrodites. On separate occasions in two bears, we recorded aberrant genitalia morphology with a high degree of clitoral hypertrophy in Svalbard, which we also classified as female pseudohermaphroditism. The observed rate of female pseudohermaphroditism in this area was 1.5% (4/269). Pseudohermaphroditism in this polar bear population could result from excessive androgen excretion by the mother caused by a tumor, or it could be a result of endocrine disruption from environmental pollutants.

Post-mortem examinations performed during May and August of 1997 on three free-living green turtles (Chelonia mydas) in the United Arab Emirates revealed that all had stomachs full of fresh seagrass (approximately 99% of the total ingesta) and presented with a duodenal volvulus involving a length of approximately 100 cm. Duodeni appeared empty and necrotic with diffuse purple-black mucosa. No apparent signs of obstruction by foreign objects, acute endoparasitism, or other disorders were observed. In all cases, duodenal volvulus was diagnosed, the cause of which may have been dietary in origin. The rise in water temperature, with an associated rise in the temperature of seagrass, thus enhancing the over-fermentation of ingesta with the subsequent liberation of excessive amounts of gas was the probable cause of volvulus formation. Ingestion of foreign bodies and physical movement also are discussed as causes of digestive disorders. It is recommended that handlers should avoid 360° rotation when overturning turtles onto their carapaces.

Two captive adult female African hedgehogs (Atelerix albiventris) had inappetance and bloody diarrhea for several days prior to death. Both hedgehogs had ulceration of the small intestine and hepatic lipidosis. Histopathology revealed small intestinal lymphosarcoma with metastasis to the liver. Extracellular particles that had characteristics of retroviruses were observed associated with the surface of some neoplastic lymphoid cells by transmission electron microscopy. These are the first reported cases of intestinal lymphosarcoma in African hedgehogs.

An adult male frilled lizard (Chlamydosaurus kingi) was found dead in Northern Territory (Australia). On physical examination it was found to be severely emaciated. At necropsy the stomach was found to be firm with mild thickening of the gastric mucosa. Gastric rugae were prominent and the mucosa was red with a thin layer of dark red mucus on its surface. Light microscopic examination revealed a mild diffuse gastritis with infiltration of the lamina propria by lymphocytes, plasma cells, and heterophils. Large numbers of small, round amphophilic to basophilic (2 to 4 μm in diameter) organisms morphologically consistent with Cryptosporidia were seen on the surface of the mucosal epithelium and free in the gastric lumen. The gastric lesions seen in this frilled lizard did not involve atrophy, as previously described in lizards with gastric cryptosporidiosis, and were similar to those described in snakes. The possibility that more than one species of Cryptosporidium parasitizes reptiles could explain the different lesions. This is the first report of cryptosporidiosis in a frilled lizard.

Serum samples from raccoons (Procyon lotor), striped skunks (Mephitis mephitis), Virginia opossums (Didelphis virginiana), and free-ranging house cats trapped in Iowa between 1984 and 1988 were tested for antibodies against Toxoplasma gondii using the modified direct agglutination test (MAT). Antibody titers ≥1:32 were considered indicative of infection. Prevalence rates by species were estimated for raccoons at 134/885 (15%), skunks at 38/81 (47%), opossums at 12/53 (23%), and cats at 16/20 (80%).

Because of their constant exposure to normal rodent definitive hosts, least weasels (Mustela nivalis) were trapped in southern Finland in late fall 1994 and examined for lung parasites. Histological examination showed that 46% of the weasels (n = 46) were infected with adiaspores identified as Chrysosporium sp. Granulomas surrounding the adiaspores consisted of mostly unorganized layers of mononuclear cells. The adiaspores from least weasels were much smaller than those reported from their prey animals. Infection with Pneumocystis carinii also was found in two weasels.

A survey of naso-pharyngeal myiasis affecting red deer (Cervus elaphus) in southern Spain was conducted. The parasites involved were the larvae of Pharyngomyia picta and Cephenemyia auribarbis (Diptera:Oestridae), which coexist sympatrically within this host. Males and older animals had higher prevalences and intensities of fly larvae. Differences in behaviour and habitat use by male and female deer, and the increase of head size in older males are possibly responsible for this. There were low densities of C. auribarbis while P. picta was the species most frequently observed, although both oestrids were located in the same host cavities. The earlier larviposition by C. auribarbis, and its faster larval development may reflect asynchronous life-cycles of both oestrids; this may decrease inter-specific competition between these sympatric species.

To determine the safety of Brucella abortus strain RB51 (SRB51) vaccine in American bison (Bison bison), 31 animals from a herd with brucellosis were used. In October 1996, 10 adult bison males and seven calves were vaccinated with the standard calfhood cattle dose of 1.8 × 10¹⁰ colony forming units (CFU) of SRB51 subcutaneously while the adult females received the standard adult cattle dose of 1 × 10⁹ CFU. Western immunoblot indicated the presence of SRB51 antibodies following vaccination. To evaluate prolonged bacterial colonization of tissues, the adult males, calves, and three adult females were divided into two groups which were slaughtered at either 13 or 16 wk post-vaccination. At necropsy, tissue samples were obtained for B. abortus culture from the liver, spleen, lymph nodes, and reproductive tract of each animal. While B. abortus field strain was cultured from one adult bull, no SRB51 was isolated from any of the animals. Seven pregnant females were monitored until parturition for signs of abortions and fetal lesions. Six cows delivered healthy calves and one delivered a dead full-term calf that was brucellae negative. Based on these results, administration of SRB51 to bison did not cause prolonged bacterial colonization of tissues in calves, adult males, or adult females. Furthermore, SRB51 did not induce abortions following vaccination in the second month of gestation.

Bacteria with inhibitory activity against Clostridium botulinum type C were isolated from 32% of sediment samples (n = 1600) collected from 10 marshes in a northern California wetland over a 12 mo period. Aerobic and anaerobic bacteria with inhibitory activity were isolated from 12% and 23% of the samples, respectively. Bacteria with inhibitory activity were isolated from all 10 study sites and throughout the year. This study demonstrates that bacteria with inhibitory activity against C. botulinum type C occur naturally in wetland sediments.

Blood samples collected from 120 wild swine (Sus scrofa) in thirteen Oklahoma (USA) counties during 1996 were tested for antibodies against six viral and two bacterial diseases. No antibodies to swine brucellosis, pseudorabies, transmissible gastroenteritis, and vesicular stomatitis were detected. Antibody titers to one or more leptospiral serovars were found in 44% of the samples, the two most frequent serovars being Leptospira interrogans serovars bratislava (29%) and pomona (27%). Antibody against porcine parvovirus and swine influenza virus was detected in 17% and 11% of the swine, respectively. Two samples (2%) were positive for antibody to the recently emerged porcine reproductive and respiratory syndrome virus.