Spirochetes were isolated from the tick, Ixodes tanuki, as well as wood mice (Apodemus speciosus) and voles (Clethrionomys rufocanus and Eothenomys smithii), caught in Fukui, Tokushima, and Hokkaido, Japan, from 1991 to 1993. Spirochetes were characterized on the basis of protein profiles, reactivities with monoclonal antibodies (mAbs), Outer surface protein A gene (ospA) and Outer surface protein B gene (ospB) amplification analysis, rRNA gene and flagellin gene restriction fragment length polymorphism (RFLP) analysis, and DNA homology values. Protein profiles of all isolates were homologous and reacted with mAb to OspA, OspB, OspC, flagellin, and heat shock protein 60. The primer reactivity to ospA and ospB were different from those of Borrelia burgdorferi sensu stricto, B. afzelii, B. japonica, and B. garinii. Based on the DNA/DNA homology value and RFLP analysis of rRNA and flagellin gene, these Borrelia sp. isolates are a new group of B. burgdorferi sensu lato. The isolates from ticks and the host rodents were identical in these assays. We propose that these Borrelia sp. are adapted to I. tanuki and are maintained in these field rodents.

Prevalence of leptospirosis was determined in California sea lions (Zalophtis californianus) stranded live along the central California (USA) coast between January 1981 and December 1994. Clinical signs of renal disease were seen in 764 (33%) of 2338 animals examined; 545 (71%) of these 764 animals died, with similar gross lesions of nephritis. In silver impregnation stains of sections of formalin-fixed kidney, numerous loosely coiled spiral organisms were observed. Leptospira pomona kenniwicki was cultured from four kidney samples in 1991. Epizootics of leptospirosis occurred in 1984, 1988, 1991, and 1994, and were more common in the autumn, typically affecting juvenile males. In 1991 and 1994, 47 animals sampled had antibody titers to L. pomona greater than 1:3200. In 1992, 20 animals sampled were seronegative, and in 1993 three of 20 animals sampled had low titers to L. pomona.

The pathogenicity of Brucella suis biovar 4 for bison (Bison bison) was evaluated by inoculation of 2.1 × 10⁷ colony forming units (CFU) in 0.1 ml saline into the conjunctival sac of six pregnant cows. Six pregnant bison were inoculated with 1.27 × 10⁷ CFU of Brucella abortus strain 2308 as a positive control. Bison were inoculated on 23 January 1992, and observed until calving or abortion after which they were euthanized, and necropsied. Bacteriological and histological examinations were conducted on lymph nodes, reproductive tract, mammary gland, and internal organs. Terminal serum samples from calves and cows were evaluated by card, rivanol precipitation, standard tube agglutination, cold complement fixation tube, indirect bison conjugated enzyme linked immunosorbent assay (ELISA), competitive ELISA, and particle-concentration fluorescence immunoassay. No clinical signs of brucellosis were seen in bison inoculated with B. suis biovar 4, and infection was found only in lymph nodes of two animals. There was no evidence of metastasis of this organism to the mammary gland or the reproductive tract. There were no detectable levels of antibodies to Brucella spp. in terminal blood samples taken from B. suis biovar 4-challenged bison. Brucella abortus was isolated from several tissues in all control bison. All B. abortus-challenged animals developed uterine infection and five developed mam-mary gland infection. Reproductive disease resulted in abortions in five B. abortus-challenged bison and neonatal death in the remaining calf. Brucella suis biovar 4 does not appear to be pathogenic for bison.

We evaluated susceptibility of Dall sheep (Ovis dalli dalli) to bacterial pneumonia induced by two strains of Pasteurella haemolytica of domestic sheep origin by evaluating the sensitivity of blood neutrophils of eight Dall sheep to lysis by cytotoxins of P haemolytica, and by intratracheal inoculation of three Dall sheep, two bighorn sheep (Ovis canadensis), and two domestic sheep with 3.7 × 10⁶ or 2.5 × 10⁷ colony forming units of P haemolytica. Neutrophils from the Dall sheep were more sensitive to lysis by cytotoxins from supernatants of a P. haemolytica, biotype A, serotype 2 (A2), of domestic sheep origin, than were neutrophils from six bighorn sheep. This cytotoxic bacterium was the same isolate that was used for intratracheal inoculation of two Dall sheep and two domestic sheep. Inoculation of this cytotoxic P. haemolytica A2 resulted in fatal fibrinopurulent pleuropneumonia in the first Dall sheep within 24 hr of inoculation, and pneumonic lesions in the second Dall sheep before it was euthanized 52 hr after inoculation. This strain of P. haemolytica A2 did not cause respiratory disease when inoculated into two domestic sheep. A noncytotoxic strain of P. haemolytica, biotype T, serotype 3,4,10 of domestic sheep origin did not result in pneumonia in the third Dall sheep or two bighorn sheep. Prior to inoculation, P. haemolytica, biotype T isolates were obtained from all three Dall sheep, but none of these isolates was cytotoxic. At necropsy, cytotoxic P. haemolytica A2 was isolated from lungs and other tissues of the two pneumonic Dall sheep. Based on these results, we conclude that Dall sheep appear to be at least as sensitive as bighorn sheep to pneumonia caused by P. haemolytica A2 of domestic sheep origin. Because in vitro and in vivo results appear closely correlated in this and other studies, we believe with additional evaluation and standardization, neutrophil cytotoxicity tests may serve as a substitute for live animal challenges in future studies of pathogenic P. haemolytica in wild sheep.

Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 P. haemolytica biotype A isolates were cytotoxic (>50% neutrophil death in vitro) to bighorn sheep neutrophils, and four of 10 P. haemolytica biotype A isolates were cytotoxic to neutrophils of cattle; P. haemolytica biotype T (= Pasteurella trehelosi) isolates were noncytotoxic to neutrophils of bighorn sheep and cattle. When six bighorn sheep were pastured with three horses, only P. haemolytica biotype T isolates were recovered from the bighorn sheep throughout the study; Pasteurella spp. organisms were not isolated from the three horses. At initiation of a study where five bighorn sheep were pastured with three cattle, P. haemolytica biotype A, serotype 1, 2 was isolated from all three cattle, and only P. haemolytica biotype T isolates were recovered from the bighorn sheep. One bighorn sheep died in each of the horse and cattle copasturing experiments. Pasteurella haemolytica was not isolated from the bighorn sheep which died in the horse copasturing experiment. A noncytotoxic P. haemolytica biotype A, serotype 2 was isolated at necropsy from the bighorn which died in the cattle contact experiment. Based on these experiments, we believe bighorn sheep and horse association would not be detrimental to bighorns due to P. haemolytica induced pneumonia.

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.

We collected 396 Gobius bucchichii, Steindachner, 1870 (Teleostei, Gobiidae) in and around the marine reserve of Cerbère-Banyuls, in the southeast of France, between March and July 1994. Five species of adult parasites were found: one acanthocephalan, Acanthocephaloides propinquus Dujardin, 1845 (Acanthocephala, Arhythmacanthidae); one nematode, Cucullanus sp. (Nematoda, Cucullanidae); and three species of digenetic trematodes, Helicometra sp. (Digenea, Opeccelidae), Derogenes sp. (Digenea, Hemiuridae) and Deretrema scorpaenicola Bartoli, 1990 (Digenea, Zoogonidae). Fishes collected in a protected area were on average, larger, older, had a higher percentage of regenerated scales, and harbored more parasites.

In the first (July 1989) of two experiments, each of three bighorn sheep (Ovis canadensis) and three domestic sheep, respectively, was exposed to 25, 150, or 300 infective third-stage larvae (L3) of the meningeal worm, Parelaphostrongylus tenuis. Two bighorn sheep had temporary mild paresis and lumbar weakness; one developed paralysis and died suddenly 32 days after exposure. Adult P. tenuis were found deep within the brain and spinal cord of the one latter sheep. A generalized inflammatory response, characterized by subdural lymphoid aggregations adjacent to spinal nerve roots, was seen in the spinal cord of most domestic and bighorn sheep. In the second experiment (September 1990), each of six domestic sheep lambs and five white-tailed deer (Odocoileus virginianus) fawns was exposed to a single dose of 15 to 125 L3 of meningeal worm. Clinical signs were seen in only one sheep; it was dull and depressed. No worms were found in this sheep. One dead adult meningeal worm was found on the brain of another sheep. First-stage larvae and adult meningeal worms were found in all deer.

In August 1994, cryptosporidiosis was diagnosed in a diarrheic fawn from a captive white-tailed deer (Odocoileus virginianus) herd maintained for research purposes at The University of Georgia's Warnell School of Forest Resources in Athens, Georgia (USA). From June through August 1995, 11 captive female white-tailed deer were housed in individual barn stalls where they gave birth to 18 fawns. Feces collected at 2 or 3 day intervals from the 18 neonatal fawns for at least 21 days and from 11 adult females once from 1 to 30 days before fawns were born and on three to 12 occasions after their birth were examined for oocysts of Cryptosporidium spp. Feces from all animals appeared normal throughout the period of examination. Oocysts morphologically indistinguishable from those of Cryptosporidium parvum were detected intermittently in the feces of one adult female from 1 to 25 days after parturition and in the feces of her fawn from 11 to 22 days of age. Oocysts also were detected intermittently in feces from twin fawns from 9 to 20 days of age, but not from their mother. Oocysts from deer were infectious for neonatal mice as determined histologically, and for calves as determined by clinical signs and excretion of oocysts.

Sera from 150 eastern barred bandicoots (Perameles gunnii) were collected from two study sites in southern Tasmania between 1992 and 1995. Samples were tested for antibodies to the protozoan parasite, Toxoplasma gondii, using formalin-treated tachyzoites as the antigen in direct (DAT) and modified agglutination tests (MAT). Cut-off titers were set based on confirmed cases of toxoplasmosis in this species. A total of 133 animals (89%) were classified as negative, seven (4.6%) had suspicious reactions, and 10 (6.7%) were diagnosed as positive. Five of the 10 positive animals were not retrapped after initial seroconversion; another three animals recorded high MAT titers on two consecutive bleedings, three months apart. Of the remaining two seropositive bandicoots, one was found dead in a trap and generalized toxoplasmosis was diagnosed at necropsy, while the other animal had central nervous system disabilities consistent with toxoplasmosis but was accidently released and never recaptured. Based on these findings we propose that eastern barred bandicoots are likely to be highly susceptible to primary T. gondii infection.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) were monitored at a Georgia (USA) site where epizootic hemorrhagic disease (EHD) and bluetongue (BT) viruses are enzootic among white-tailed deer (Odocoileus virginianus). Collections were made using a captive white-tailed deer and light traps from June 1993 through November 1994. We collected 210,482 females from the captive deer during morning and evening periods. Predominant species were C. lahillei (73%), C. stellifer (16%), C. biguttatus (6%), C. niger (3%), C. spinosus (2%), and C. paraensis (0.2%). Other species were C. venustus, C. obsoletus/sanguisuga, C. haematopotus, C. guttipennis, and C. arboricola, which together represented <0.1% of the specimens collected. No C. variipennis, a known vector of EHD and BT viruses, were collected from the deer. An estimated 953,299 females were collected in 695 light-trap nights. The most common species in light-trap collections were C. spinosus (45%), C. biguttatus (27%) and C. stellifer (24%). Culicoides variipennis was rare in the light-trap samples, representing <0.01% of the total collections. There was serological evidence from hunter-killed deer that local deer were infected with EHD and BT viruses during the study, particularly during 1994. A primary suspect vector was C. lahillei, which attacked the bait deer in large numbers during the summer and early fall of both 1993 and 1994. Based on their seasonality, relative abundance, and host-seeking activity, C. stellifer and C. spinosus also were considered as possible vectors. However, virus isolation attempts on 113,716 Culicoides, including 62,530 C. lahillei and 32,769 C. stellifer, were negative.

I established reference values for weight, hematology, and serum chemistry for seven species of free-ranging Hawaiian tropical pelagic seabirds comprising three orders (Procellariiformes, Pelecaniformes, Charadriiformes) and six families (Procellariidae, Phaethontidae, Diomedeidae, Sulidae, Fregatidae, and Laridae). Species examined included 84 Hawaiian dark-rumped petrels (Pterodoma phaeopygia), 90 wedge-tailed shearwaters (Puffinus pacificus), 151 Laysan albatrosses (Diomedea immutabilis), 69 red-footed boobies (Sula sula), 154 red-tailed tropicbirds (Phaeton rubricauda), 90 great frigatebirds (Fregata minor), and 72 sooty terns (Sterna fuscata). Hematocrit, total plasma solids, total and differential white cell counts, serum glucose, calcium, phosphorus, uric acid, total protein, albumin, globulin, aspartate aminotransferase and creatinine phosphokinase were analyzed. Among and within species, hematology and chemistry values varied with age, sex, season, and island of collection. Despite this variation, order-wide trends were observed.

An adult greater white-toothed shrew (Crocidura russula) had both a quadricuspid aortic valve and a single coronary artery arising from the aorta. The shrew was caught on 10 May 1994 in the environs of Málaga, southern Spain. Both congenital anomalies may be potential causes of cardiac dysfunction, but apparently produced no significant cardiac complication in the shrew. This is the first report of a quadricuspid aortic valve in a wild-living mammal.

Retained testes were found in 11 (16%) of 71 black bears (Ursus americanus) examined over a 3-year period in Florida (USA). Four of the 11 bears were older than one year and weighed more than 32 kg; therefore, they were considered to be cryptorchid. The remaining seven bears may have had delayed testicular descent due to their apparent normal immature development. This is the first known published report of the prevalence of cryptorchidism and apparently normal delayed testicular descent in a black bear population.

Between February 1992 and March 1994, four species of rodent from the Snake River Birds of Prey Area near Boise, Idaho (USA) were necropsied. Hemorrhagic gastritis was observed in 16 of 131 Townsend's ground squirrels (Spermophilus townsendii), one of 11 Ord's kangaroo rats (Dipodomys ordii) and the one Great Basin pocket mouse (Perognathus parvus) evaluated. No lesions were observed in 14 white-footed deer mice (Peromyscus maniculatus). Tissue from one Townsend's ground squirrel was negative for Helicobacter sp.-like bacteria.

A case of lymphosarcoma in a captive adult female raccoon (Procyon lotor) from northeastern Pennsylvania (USA) was observed in 1991. Prior to its death the raccoon had lost weight. At necropsy the carcass was in poor body condition and had pale mucous membranes. The thoracic and abdominal lymph nodes were enlarged, soft, and pale tan. Microscopically, there was effacement of normal lymph node architecture by sheets of mononuclear cells. These were well-differentiated small lymphocytes with distinct cell borders. Nuclei of these cells were darkly stained and mitotic figures were frequently seen. Similar but lesser numbers of neoplastic cells were seen in the parenchyma of liver, spleen, and the pancreas. Since the neoplasm involved several organs, we propose that the condition was of multicentric origin. Gross lesions, histopathologic findings and the organs involved differed from a previously described case of lymphosarcoma in a raccoon.

During March 1988, a case of bilateral polycystic kidney disease (PKD) occurred in an aged raccoon (Procyon lotor) at a zoo in Wilmington, Delaware (USA). Prior to its death, the raccoon had no clinical signs. On necropsy there was bilateral enlargement of kidneys which, on cut sections, had many variable sized fluid-filled cystic cavities. Endometrial hyperplasia with presence of multiple variable-sized cysts were also seen in the uterus of this raccoon. Microscopical examinations were characteristic of an end-stage renal failure due to PKD. Neither PKD nor cystic endometerial hyperplasia appears to have been previously described in this species.

To investigate a possible link of malignant hyperthermia to capture myopathy, between June 1990 and July 1993 we anesthetized four black-tailed deer (Odocoileus hemionus columbianus) and challenged them with halothane and succinylcholine. Halothane had no significant effect on oxygen consumption. Succinylcholine significantly (P < 0.05) increased cardiac output (mean ± SD), from 2.94 ± 1.05 1/min to 5.26 ± 1.79 1/min, and oxygen consumption, from 5.5 ±2.1 ml/kg/min to 10.1 ± 2.9 ml/kg/min. Muscle biopsy specimens tested for malignant hyperthermia susceptibility responded normally to halothane and caffeine. We conclude that these deer did not experience malignant hyperthermia, suggesting no link to capture myopathy.

A free-living adult male gopher tortoise (Gopherus polyphemus) was found on Sanibel Island, Florida (USA), on 18 February 1992 with signs of upper respiratory disease. On necropsy after euthanasia on 27 February 1992, severe, extensive necrotizing ulcerative tracheitis, multifocal necrotizing pneumonia, and multifocal necrotizing ulcerative pharyngitis and esophagitis were observed. Large ovoid to round intracytoplasmic basophilic inclusions, which appeared to displace the nucleus to the cell periphery, occurred within degenerate and necrotic epithelial cells of the above tissues. On transmission electron microscopy of formalin-fixed trachea and lung, intracytoplasmic viral particles were observed within necrotic cells in the tracheal lumen and epithelial cells of the lung. Most infected cells also had a roughly spherical granular cytoplasmic inclusion that contained clusters of viral particles. Viral particles had an electron dense spherical to icosahedral core surrounded by a less electron dense icosahedral capsid. Mature extracellular virions were surrounded by an envelope and were 150 to 220 nm in diameter. Virions and cytoplasmic inclusions were morphologically similar to those of the Family Iridoviridae.

Muskrat (Ondatra zibethicus) immunoglobulin fraction was separated from whole serum by Protein A Sepharose chromatography. In serum electrophoresis, this fraction had a gamma motility; when electrophoresed on a polyacrylamide gel with sodium dodecyl sulfate it resolved into two protein bands of approximately 52 and 25 kilodaltons, respectively These bands were consistent with molecular weights of known heavy and light chains of immunoglobulin G (IgG) in other closely related species. Furthermore, the putative muskrat immunoglobulins had a strong cross-reactivity with mouse IgG₁, IgG₃, and kappa chain in an enzyme-linked immunosorbent assay. We propose, that the proteins bound to the Protein A Sepharose represent muskrat immunoglobulins of the IgG class.

We report cytologic observations and isolation of Mycoplasma felis in September 1992 from the lower respiratory tract of a 3-week-old captive serval (Felis serval) cub with pneumonia, in Florida (USA). Septic, neutrophilic inflammation with a large, monomor-phic population of unique, pleomorphic, intracellular and extracellular rods was diagnosed from a transtracheal wash. Mycoplasma felis was the only bacterium isolated in significant numbers from the transtracheal wash.

Although antibodies to viruses in both the epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) serogroups 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.

Meningeal worms (Parelaphostrongylus tenuis) were found in each of five white-tailed deer (Odocoileus virginianus) examined from Wassaw Island, Chatham County, Georgia, in September 1993. This represents the first reported occurrence of the parasite on a southeastern barrier island and extends its geographic distribution approximately 140 km beyond the nearest known infected mainland deer population. According to an anecdotal account, six white-tailed deer were imported from Pennsylvania and released on Wassaw Island in 1905 or shortly thereafter. Based on its absence elsewhere along the southeastern coast from North Carolina to Louisiana and its high prevalence in Pennsylvania, the enzootic focus of P. tenuis on Wassaw Island was attributed to translocation of infected deer.

During necropsy of a Burton's snake lizard (Lialis burtonis Gray 1835) that was imported from Australia in January 1994, juvenile acanthocephalans were recovered from the mesentery and small bowel serosa which were identified as Porrorchis hylae (Johnston, 1914). This find represents a new paratenic host record for the species, and is recorded in a reptile native to Australia for the first time.

Coccidioidomycosis is described in seven California sea lions (Zalophus californianus) admitted to The Marine Mammal Center, Sausalito, California (USA), between January 1986 and December 1994. Diagnoses were confirmed by histology in all seven cases, culture in three cases, and serology in one case. These are believed to be the first published cases of coccidioidomycosis in free-ranging California sea lions.

We determined that antibody responses to Psoroptes sp. mites were not present in 403 of 407 sera samples collected opportunistically from 1979 through 1991 from Dall sheep (Ovis dalli) from five locations in Alaska, USA (Eastern Arctic, n = 61; Central Arctic, n = 15; Western Interior, n = 122; Central Interior, n = 63; Eastern Interior, n = 146). Test values for four samples exceeded the positive cutoff value for the immunoassay, but exposure to mites could not be confirmed since the 95% confidence interval for true prevalence ranged from 0 to 2.3%. Therefore, we concluded that these were probably false positive results. Our analysis, coupled with the lack of previous reports of mites or lesions in Dall sheep or other Alaskan ungulates, provided indirect evidence that Psoroptes sp. are not enzootic in Dall sheep in Alaska. In contrast, Psoroptes sp. have been reported in bighorn sheep (Ovis canadensis) and other wild ungulate populations from southern Canada to Mexico. These findings are compatible with the hypothesis that Psoroptes sp. were introduced into North America with imported domestic sheep and were not introduced by ancestral wild sheep.

We tested the null hypothesis that anesthetizing meadow voles (Microtus californicus) and brushing them vigorously for ectoparasites would have no effect on the later recapture of these voles. Voles were trapped from 6 December 1993 to 10 January 1994 at Faye Slough Wildlife Area near Eureka, Humboldt County, California (USA). Alternate trapped voles were anesthetized with ethyl ether and brushed vigorously for ectoparasites. There was no significant difference in the frequency of recapture nor in the time to first recapture between those voles anesthetized and brushed, compared to control animals.