We studied the pathology, epidemiology, and clinical significance of genital herpesvirus infection in a zoo collection of bottlenose dolphins (Tursiops truncatus). Samples from the genital mucosa of male (n=21) and female (n=15) dolphins were tested by nested polymerase chain reaction (PCR) targeting the DNA polymerase of herpesvirus. Herpesvirus infection was significantly associated with the occurrence of mucosal plaques on penis (n=3) or vulva (n=4). Biopsies from a penile plaque showed epithelial hyperplasia by histology, contained herpesvirus-like particles by electron microscopy, and tested positive for herpesvirus by PCR. Herpesvirus was successfully cultivated from penile plaque samples and identified as a member of the Gammaherpesvirinae by DNA sequencing and phylogenetic analysis. We used the newly cultivated bottlenose dolphin herpesvirus (TTHV) to develop a direct enzyme-linked immuno-sorbent assay for anti-TTHV antibodies in banked sera of these dolphins. The percentage of positive samples was higher in adults (20/21, 95%) than in juveniles (7/15, 47%). Seroconversion occurred around the age of onset of sexual behavior. Although herpesvirus infection has been associated with abortion, perinatal mortality, and urogenital neoplasia in other species, we found no evidence of herpesvirus infection by PCR in tissues from six cases of abortion and perinatal mortality, and no diagnoses of urogenital tumors in 24 bottlenose dolphins from this zoo collection that died since 1990. Together, we here report the first successful cultivation from bottlenose dolphins of a herpesvirus that probably causes benign genital plaques, is endemic in this group of dolphins, and is likely transmitted by sexual contact.

Madagascar has long been recognized for its unique and diverse biota. In particular, significant effort has been made to establish baseline population data to better conserve the endemic avifauna. During field expeditions between 1993 and 2004, birds were mist-netted at 11 different sites, at elevations from 60 m to 2,050 m above sea level. Data on endemic status, forest type, and habitat preference were recorded. Thin blood films from 947 birds, belonging to 26 families and 64 species, were examined by light microscopy to determine the prevalence of blood parasites. Of these 947 birds, 30.7% were infected by at least one species of blood parasite, 26.8% of which were infected by more than one species. Species of Haemoproteus were the most prevalent (17.4%), followed by microfilariae (11.0%), Leucocytozoon spp. (9.4%), Plasmodium spp. (1.9%), Trypanosoma spp. (0.9%), and Babesia spp. (0.2%). Species level identifications confirmed the presence of 47 species of hemosporidians and trypanosomes, which is notably high and mirrors the diversity of their avian hosts. Eleven (23.4%) of these parasite species were new to science and thought to be endemic to the island. Significant differences in prevalence were observed by sample site, forest type (humid vs. dry), and habitat preference. Birds from all elevational zones sampled were infected, although not all parasite genera were present in each zone. Four of the six endemic avian families or subfamilies (Bernieridae, Brachypteraciidae, Philepittinae [Eurylaimidae], and Vangidae) were sampled and found to be parasitized. Of the families with the largest sample sizes, the Zosteropidae and Ploceidae had the highest prevalence of infection (65.6% and 49.3%, respectively). The vectors of hematozoan parasites in Madagascar are currently unknown. These results add to the current knowledge of avian parasitism in Madagascar and are of particular interest for the conservation of endemic species, as well as threatened or endangered populations.

Spargana of Spirometra erinacei infect many vertebrate species, but severe disease from sparganosis has been reported from few host species. Information on the effects of this common, introduced tapeworm of cats on Australian frogs is lacking. Our survey to detect significant diseases in free-ranging amphibians in eastern Australia between 1993 and 2000 revealed that infection with spargana (plerocercoids) of S. erinacei occurred in 12/243 (4.9%) sick frogs. Infections occurred in skeletal muscle and subcutis, especially the thighs, of large adults of Litoria caerulea, Litoria aurea, Litoria gracilenta, and Litoria peronii. Three frogs were also infected in the coelomic cavity. Heavy burdens in seven frogs were associated with poor body condition and debilitating lesions, whereas lighter infections in five sick frogs were considered likely to be incidental to other diseases. In severe infections, a large proportion of thigh muscle was replaced with spargana and various amounts of fibrosis, and some frogs also had myonecrosis, granulomatous inflammation, hemorrhage, and skin ulceration. Concurrent infections were common. Our findings suggest sparganosis is one of a few currently recognized serious diseases affecting free-ranging frogs in Australia.

The prevalence and phenotypic variability of Pasteurella and Mannheimia isolates from Sierra Nevada bighorn sheep (Ovis canadensis sierrae), White Mountain bighorn sheep (Ovis canadensis nelsoni), and domestic sheep (Ovis aries) from California, USA, were compared. The White Mountain bighorn sheep population had a recent history of pneumonia-associated mortality, whereas the Sierra Nevada bighorn sheep population had no recent history of pneumonia-associated mortality. The domestic sheep flocks were pastured in areas geographically near both populations but were not known to have direct contact with either bighorn sheep population. Oropharyngeal swab samples were collected from healthy domestic and bighorn sheep and cultured to characterize bacterial species, hemolysis, biogroups, and biovariants. Pasteurella trehalosi and Mannheimia haemolytica were detected in all of the study populations, but the relative proportion of each bacterial species differed among sheep populations. Pasteurella trehalosi was more common than M. haemolytica in the bighorn sheep populations, whereas the opposite was true in domestic sheep. Mannheimia haemolytica was separated into 11 biogroups, and P. trehalosi was characterized into two biogroups. Biogroup distributions for M. haemolytica and P. trehalosi differed among the three populations; however, no difference was detected for the distribution of P. trehalosi biogroups between the Sierra Nevada bighorn sheep and domestic sheep. The prevalence odds ratios (pOR) for the distribution of M. haemolytica biogroups suggested little difference between White Mountain bighorn sheep and domestic sheep compared with Sierra Nevada bighorn sheep and domestic sheep, although these comparisons had relatively large confidence intervals for the point estimates. Hemolytic activity of the isolates was not different among the sheep populations for M. haemolytica but was different for P. trehalosi. No clear evidence of association was found in the Pasteurellaceae distribution between the White Mountains bighorn sheep and domestic sheep.

The javelina, or collared peccary (Pecari tajacu), is indigenous to Arizona, New Mexico, and Texas in the United States and ranges throughout Latin America. From June 2004 to April 2005, an estimated 105 javelinas died in a mortality event that occurred in Tucson, Arizona, and neighboring areas. Clinical signs observed in sick animals included emaciation, dehydration, lethargy, and diarrhea. In addition, some animals showed labored breathing and hind limb weakness. We necropsied 34 animals, and enteritis was the most frequent clinical sign, followed by colitis, pulmonary congestion, and pneumonia. The only consistent findings were isolations of Clostridium perfringens type A and multiple Salmonella serotypes. Although it is likely that these javelinas ultimately succumbed to salmonellosis, it is unclear whether other unidentified underlying factors were involved. This is the first reported case of widespread salmonellosis in free-ranging javelinas.

Bats are one of the most widely distributed mammals in the world, and they are reservoirs or carriers of several zoonoses. Bats were trapped in 27 geographic locations across Trinidad and Tobago, and following euthanasia, gastrointestinal tracts were aseptically removed. Contents were subjected to bacteriologic analysis to detect Salmonella spp., Escherichia coli, and Campylobacter spp. Isolates of Salmonella were serotyped, and E. coli isolates were screened for O157 strains and antimicrobial sensitivity to eight antimicrobial agents; phenotypic characteristics also were determined. Of 377 tested bats, representing 12 species, four bats (1.1%) were positive for Samonella spp, 49 (13.0%) were positive for E. coli, and no bats were positive for E. coli O157 strain or Campylobacter spp. Isolated serotypes of Salmonella included Rubislaw and Molade, both from Noctilio leporinus, a fish-eating bat, Caracas recovered from Molossus major, and Salmonella Group I from Molossus ater, both insect-eating bats. Of the 49 isolates of E. coli tested, 40 (82%) exhibited resistance to one or more antimicrobial agents, and the prevalence of resistant strains was comparatively high to erythromycin (61%) and streptomycin (27%) but lower to gentamycin (0%) and sulphamethozaxole/trimethoprim (2%).

In an attempt to identify critical health issues affecting the survival of endangered leatherback sea turtles (Dermochelys coriacea), a prospective study was conducted in several dead-in-nest hatchlings and captive posthatchlings to examine pathologic changes and presence of pathogenic microorganisms. Numerous histopathologic changes were identified. Although bacterial etiologies were suspected in deaths of captive individuals, a single causative organism was not identified but rather, a mixed population of bacterial flora was cultured. Muscle degeneration observed in most samples implicates a potential environmental factor in species survival and needs future investigation.

This study evaluated the health status and established hematologic and serum biochemistry parameters for free-living nestlings of the Hyacinth Macaw (Anodorhynchus hyacinthinus) from the Brazilian Pantanal (19°51′–19°58′S; 56°17′–56°24′W), for four consecutive years (from December 2003 through December 2006). Physical examinations indicated that all the birds were in good health. Endoparasites and blood parasites were not detected in any of the nestlings, and ectoparasites seemed to be limited to Philornis sp. (Diptera: Muscidae). Significantly higher levels of total white blood cells and heterophils, glucose, total protein, triglycerides, and phosphorus were observed in females. In females, higher cholesterol levels and packed cell volumes were observed in older birds, and total white blood cell and heterophil counts were higher in young animals. In males, uric acid levels were higher in older individuals. Wild Pantanal Hyacinth Macaws feed on only two species of palm nuts (Acrocomia totai and Scheelea phalerta). This limited food habit has a strong impact on population size and may alter the clinical pathology parameters of these birds. Therefore, knowledge of blood levels in normal individuals is essential to assess the physiologic and pathologic condition of wild macaws, to assess the effects of environmental changes on their health, and to contribute to conservation strategies of this endangered species.

We necropsied a moribund, wild adult male desert tortoise (Gopherus agassizii) with clinical signs of respiratory disease and elevated plasma biochemical analytes indicative of renal disease (blood urea nitrogen [415 mg/dl], uric acid [11.8 mg/dl], sodium >180 mmol/l] and chloride [139 mmol/l]). Moderate numbers of birefringent oxalate crystals, based on infrared and electron microscopy, were present within renal tubules; small numbers were seen in colloid within thyroid follicles. A retrospective analysis of 66 additional cases of wild desert tortoises was conducted to determine whether similar crystals were present in thyroid and kidney. The tortoises, from the Mojave and Sonoran deserts, were necropsied between 1992 and 2003 and included juveniles and adults. Tortoises were classified as healthy (those that died due to trauma and where no disease was identified after necropsy and evaluation by standard laboratory tests used for other tortoises) or not healthy (having one or more diseases or lesions). For all 67 necropsied tortoises, small numbers of crystals of similar appearance were present in thyroid glands from 44 of 54 cases (81%) and in kidneys from three of 65 cases (5%). Presence of oxalates did not differ significantly between healthy and unhealthy tortoises, between age classes, or between desert region, and their presence was considered an incidental finding. Small numbers of oxalate crystals seen within the kidney of two additional tortoises also were considered an incidental finding. Although the source of the calcium oxalate could not be determined, desert tortoises are herbivores, and a plant origin seems most likely. Studies are needed to evaluate the oxalate content of plants consumed by desert tortoises, and particularly those in the area where the tortoise in renal failure was found.

Chronic wasting disease (CWD), a prion disease affecting North American cervids, has been discovered in at least 12 states and provinces throughout the continent. Since 2002, a number of states and provinces have initiated surveillance programs to detect CWD in native cervid populations. However, many questions remain about the appropriate methods, geographic scope, and number of samples required for an effective CWD surveillance program. We provide an improved statistical method to calculate the probability of detecting CWD in primary sample units (e.g., county or deer management unit) that also considers deer abundance and the nonrandom distribution of CWD and hunter harvests. We used this method to analyze data from a statewide CWD detection program conducted in Wisconsin during the autumns of 2002 and 2003 to determine the distribution of CWD in white-tailed deer (Odocoileus virginianus). Deer heads were collected at hunter registration stations, and brainstem (obex) and retropharyngeal lymph nodes were removed for disease testing. Our analysis includes samples from >35,000 deer collected outside the known affected area. The probability of detecting chronic wasting disease at a prevalence of 1% varied from 0.89 to ≥0.99 among the 56 primary sample units. Detection probabilities for 1% CWD prevalence were >0.9 in 55 primary sample units, and >0.99 in 10. Detection probabilities will be higher in areas where CWD prevalence exceeds 1%. CWD-positive deer were detected in eight primary sample units surrounding the known affected area during surveillance activities. Our approach provides a novel statistical technique to accommodate nonrandom sampling in wildlife disease surveillance programs.

We examined seasonal dispersal patterns and timing of new infections of Sin Nombre virus (SNV), as determined by recent acquisition of antibodies (seroconversion), in deer mice (Peromyscus maniculatus) at two Montana rangeland study sites over three years, 2004–2007. One study site was located in grassland habitat, and the other was located in shrub-steppe. In Montana, both of these habitats are commonly associated with peridomestic environments (in and around buildings). Peridomestic environments are where most reported human cases of hantavirus pulmonary syndrome (HPS) likely originate. Furthermore, deer mice dispersing from sylvan habitats colonize peridomestic environments. Thus, a thorough understanding of deer mouse dispersal is needed to help predict when humans are most at risk for exposure to SNV. We trapped mice at each study site twice a month, accumulating 85,200 trap nights of effort and capturing 6,185 individual deer mice a total of 22,654 times. We documented 980 dispersing individuals over 3 yr. We found positive correlations between the number of dispersing mice and number captured at each site, but there were no statistically significant seasonal differences in the number of dispersing mice. However, we did find a spring/summer bias in mice that seroconverted and dispersed, suggesting that recently infected deer mice are most likely to enter settings where humans may be exposed to SNV during spring and summer.

Severe population declines have resulted from the spillover of canine distemper virus (CDV) into susceptible wildlife, with both domestic and wild canids being involved in the maintenance and transmission of the virus. This study (March 2001 to October 2003) collated case data, serologic, pathologic, and molecular data to describe the spillover of CDV from domestic dogs (Canis familiaris) to black-backed jackals (Canis mesomelas) during an epidemic on the Namibian coast. Antibody prevalence in jackals peaked at 74.1%, and the clinical signs and histopathologic observations closely resembled those observed in domestic dog cases. Viral RNA was isolated from the brain of a domestic dog from the outbreak area. Sequence data from the phosphoprotein (P) gene and the hemagglutinin (H) genes were used for phylogenetic analyses. The P gene sequence from the domestic dog shared 98% identity with the sequence data available for other CDV isolates of African carnivores. For the H gene, the two sequences available from the outbreak that decimated the lion population in Tanzania in 1994 were the closest match with the Namibian sample, being 94% identical across 1,122 base pairs (bp). Phylogenetic analyses based on this region clustered the Namibian sample with the CDV that is within the morbilliviruses. This is the first description of an epidemic involving black-backed jackals in Namibia, demonstrating that this species has the capacity for rapid and large-scale dissemination of CDV. This work highlights the threat posed to endangered wildlife in Namibia by the spillover of CDV from domestic dog populations. Very few sequence data are currently available for CDV isolates from African carnivores, and this work provides the first sequence data from a Namibian CDV isolate.

Bison (Bison bison) and elk (Cervus elaphus nelsoni) in the Greater Yellowstone Area (GYA), USA, are infected with Brucella abortus, the causative agent of bovine brucellosis, and they serve as a wildlife reservoir for the disease. Bovine brucellosis recently has been transmitted from infected elk to cattle in Montana, Wyoming, and Idaho and has resulted in their loss of brucellosis-free status. An efficacious Brucella vaccine with a delivery system suitable for wildlife would be a valuable tool in a disease prevention and control program. We evaluated Strain 19 (S19) in a sustained release vehicle consisting of alginate microspheres containing live vaccine. In a challenge study using red deer (Cervus elaphus elaphus) as a model for elk, alginate, a naturally occurring polymer combined with a protein of Fasciola hepatica vitelline protein B was used to microencapsulate S19. Red deer were orally or subcutaneously immunized with 1.5×10¹⁰ colony-forming units (CFUs) using microencapsulated S19. Humoral and cellular profiles were analyzed bimonthly throughout the study. The vaccinated red deer and nonvaccinated controls were challenged 1 yr postimmunization conjunctivally with 1×10⁹ CFUs of B. abortus strain 2308. Red deer vaccinated with oral microencapsulated S19 had a statistically significant lower bacterial tissue load compared with controls. These data indicate for the first time that protection against Brucella-challenge can be achieved by combining a commonly used vaccine with a novel oral delivery system such as alginate-vitelline protein B microencapsulation. This system is a potential improvement for efficacious Brucella-vaccine delivery to wildlife in the GYA.

In 2003–06, targeted (active) surveillance for European bat lyssaviruses (EBLVs) was undertaken throughout England, focusing on two species most likely to host these viruses, Myotis daubentonii and Eptesicus serotinus. Blood was sampled for the detection of EBLV-specific neutralizing antibodies and oropharyngeal swabs were taken for the detection of viral RNA or infectious virus in saliva. Between 2003 and 2006, 273 E. serotinus and 363 M. daubentonii blood samples were tested by the EBLV-1 or EBLV-2 specific modified fluorescent antibody neutralization test. The EBLV-2 antibody prevalence estimate was 1.0–4.1% (95% confidence interval [CI]; mean=2.2%) for M. daubentonii. European bat lyssavirus type 1–specific antibodies were detected only in a single E. serotinus. Other nontarget species (n=5) were sampled in small numbers (n=24), with no EBLV-specific antibody detected. No viral RNA or live virus was detected in any of the oropharyngeal swabs analyzed. Host RNA was detected from 83% of the oropharyngeal swabs analyzed (total swabs 2003–06: n=766). These data show that EBLV-2 is present in M. daubentonii in England. In contrast, there is insufficient evidence to suggest that EBLV-1 is present in E. serotinus in England, although further research is warranted.

The eradication of tuberculosis (Mycobacterium bovis infection) from cattle herds may be compromised if infected wildlife species, such as European badgers (Meles meles), share the same environment and contribute to transfer of infection. Options for dealing with tuberculosis in this wild reservoir host are limited by conservation and social concerns, despite a clear implication that infected badgers are involved with the initiation of tuberculosis in cattle herds. Vaccination of badgers against M. bovis, if successfully employed, would directly facilitate the completion of bovine tuberculosis eradication in affected areas. Vaccine trials in captive badgers have established that the M. bovis bacille Calmette-Guérin (BCG) vaccine can induce a protective response that limits the distribution and severity of tuberculosis disease following experimental challenge. The protective effect of the vaccine has been demonstrated when the vaccine was delivered by subcutaneous injection, deposited on mucous membranes, and given orally in a lipid formulation. A large-scale field trial of oral BCG vaccine has been designed to measure the protection generated in wild badgers subjected to natural transmission of infection and to estimate vaccine efficacy. These parameters will be estimated by comparing the prevalence of M. bovis infection in vaccinated and nonvaccinated badgers. The results will provide a framework for the development and implementation of a national strategy to eliminate the disease in badger populations and if successful will remove this major impediment to bovine tuberculosis eradication.

Tuberculosis has been diagnosed in wild boar (Sus scrofa) in several European countries during the last decade; however, almost no information has been reported to date for Portugal. This study aimed to investigate tuberculosis in wild boar in Portugal through characterization of Mycobacterium bovis infection and identification of disease risk factors. Tissue samples were obtained from hunted wild boar during the 2005 and 2006 hunting seasons. Samples were inspected for gross lesions and processed for culture. Acid-fast bacterial isolates were identified by polymerase chain reaction and spoligotyping. Associations between tuberculosis in wild boar and several variables linked to wild ungulate diversity and relative abundance, livestock density, and cattle tuberculosis incidence were investigated. Mycobacterium bovis isolates were identified in 18 of 162 wild boars from three of eight study areas. Infection rates ranged from 6% (95% confidence interval [CI_P95%]=1–21%) to 46% (CI_P95%=27–67%) in the three infected study areas; females in our sample were at greater risk of being infected than males (odds ratio=4.33; CI_P95%=3.31–5.68). Spoligotyping grouped the M. bovis isolates in three clusters and one isolate was a novel spoligotype not previously reported in international databases. Detection of M. bovis was most consistently associated with variables linked to wild ungulate relative abundance, suggesting that these species, particularly the wild boar, might act as maintenance hosts in Portugal.

Bovine tuberculosis (TB) is an important economic disease. Badgers (Meles meles) are the wildlife source implicated in many cattle outbreaks of TB in Britain, and extensive badger control is a controversial option to reduce the disease. A badger and cattle population model was developed, simulating TB epidemiology; badger ecology, including postcull social perturbation; and TB-related farm management. An economic cost-benefit module was integrated into the model to assess whether badger control offers economic benefits. Model results strongly indicate that although, if perturbation were restricted, extensive badger culling could reduce rates in cattle, overall an economic loss would be more likely than a benefit. Perturbation of the badger population was a key factor determining success or failure of control. The model highlighted some important knowledge gaps regarding both the spatial and temporal characteristics of perturbation that warrant further research.

Surveillance approaches for wildlife diseases often are based on strategies devised for livestock diseases. Following standard protocols, surveillance sometimes continues after apparent disease elimination. However, in the case of recurrent wildlife diseases that cause decisive morbidity and mortality, efficient and effective surveillance strategies might need to be more dynamic and adaptable to the actual epidemic situation. Here, we evaluated existing surveillance schemes by reanalyzing historic data on three wildlife diseases in Europe: rabies, classical swine fever, and avian influenza. We analyzed the aims of different surveillance activities and the way in which they were performed. Our analyses revealed that static, nonadaptive surveillance was a suboptimal approach. Consequently, we propose and discuss a more adaptive alternative scheme of situation-based surveillance for recurrent wildlife diseases that cause readily recognizable morbidity and mortality.

Wild deer populations in Great Britain are expanding in range and probably in numbers, and relatively high prevalence of bovine tuberculosis (bTB, caused by infection with Mycobacterium bovis) in deer occurs locally in parts of southwest England. To evaluate the M. bovis exposure risk posed to cattle by wild deer relative to badgers in England and Wales, we constructed and parameterized a quantitative risk model with the use of information from the literature (on deer densities, activity patterns, bTB epidemiology, and pathology) and contemporary data on deer, cattle, and badger (Meles meles) distribution and abundance. The median relative risk score for each of the four deer species studied—red (Cervus elaphus), fallow (Dama dama), and roe (Capreolus capreolus) deer, and muntjac (Muntiacus reevesi)—was lower than unity (the relative risk set for badgers, the putative main wildlife reservoir of M. bovis in England and Wales). However, the 95th percentiles associated with risk estimates were large, and the upper limits for all four deer species exceeded unity. Although M. bovis exposure risks to cattle from deer at pasture are likely to be lower than those from badgers across most areas of England and Wales where cattle are affected by bTB because these areas coincide with high-density badger populations but not high-density deer populations, we predict the presence of localized areas where relative risks posed by deer may be considerable. Moreover, wherever deer are infected, risks to cattle may be additive to those posed by badgers. There are considerable knowledge gaps associated with bTB in deer, badgers, and cattle, and data available for model parameterization were generally of low quality and high variability, and consequently model output were subject to some uncertainty. Improved estimates of the proportion of time that deer of each species spend at pasture, the likelihood and magnitude of M. bovis excretion, and local badger and deer densities appear most important for improving estimates of relative risk in this system.

The red fox (Vulpes vulpes) is the main terrestrial wildlife rabies vector in Europe. However, recently the raccoon dog (Nyctereutes procyonoides)—an invasive species originating from East Asia—has become increasingly important as secondary host, especially in the Baltic States. This imposes problems on neighboring rabies-free countries (such as Finland), where the density of each of the two vector species on its own might be too low to sustain a long-term rabies epizootic, but the community of vectors could be large enough to support a rabies epizootic. In this modeling study, we analyzed rabies epizootics in a community of foxes and raccoon dogs. We focused on the impact of density and behavioral differences (hibernation) between the two vector species. We found that rabies could persist in the community, even if the disease would not spread in the single vector species because its density was too low. Epizootics in the community were stronger than expected for single species, and raccoon dogs were usually the major rabies host. If raccoon dog territory density was high, invasive raccoon dogs could even outcompete native foxes because of apparent competition via the rabies virus. The enhancement in disease risk and disease intensity caused by raccoon dogs suggests that current strategies to control wildlife rabies in Europe should be reviewed, and that oral rabies vaccination also should target raccoon dogs after they emerge from hibernation.

Estimates of chronic disease prevalence are needed to improve our understanding of marine disease epizootiology, which is poorly known for marine megafauna such as marine turtles. An emerging worldwide threat to green sea turtles (Chelonia mydas) is fibropapillomatosis (FP), which is a pandemic tumor-forming disease associated with herpesviruses. We report on a 26-yr FP epidemic in the Hawaiian Archipelago and show that apparent disease prevalence in the world's main endemic hot spot increased rapidly following a late 1980s outbreak, peaked during the mid-1990s, and then declined steadily ever since. While this disease is a major cause of sea turtle stranding in Hawaiian waters and can be fatal, we also show that long-term tumor regression can occur even for turtles with advanced FP. The endemic Hawaiian green turtle stock was severely depleted by overexploitation prior to protection under the US Endangered Species Act in 1978. This stock has increased significantly ever since, despite exposure to a major chronic disease epidemic that is currently declining.

There are two recognized poxviruses that are associated with disease in tree squirrels: squirrel fibroma virus (SQFV), Leporipoxvirus, which affects eastern grey squirrels (Sciurus carolinensis) in eastern North America, and squirrelpox virus (SQPV), a member of a newly identified poxvirus genus, which affects European red squirrels (Sciurus vulgaris) in the United Kingdom. In August 2008, a cutaneous poxvirus-associated disease was identified in a North American red squirrel (Tamiasciurus hudsonicus) from the Yukon Territory, Canada. The gross and microscopic appearance of the skin lesions was more consistent with SQPV than SQFV, and electron microscopy revealed poxvirions only within epithelial cells. Polymerase chain reaction (PCR) was used to identify poxvirus core protein encoding DNA in skin samples, and phylogenetic analysis showed that the inferred amino acid sequence was distinct from all other poxvirus species for which the core protein gene has been sequenced, including those of the genus Leporipoxvirus. Although the core protein sequence of SQPV was not available, comparison of the constructed phylogenetic tree to other published trees, based on major outer envelope proteins, revealed that the identified sequence occupies a position similar to SQPV in terms of its relationship to other poxviruses. However, PCR primers designed to amplify gene sequences encoding the SQPV major envelope protein and RNA polymerase did not amplify any sequences from infected tissues. These findings suggest that the virus present in this squirrel is a novel poxvirus of North American red squirrels. To our knowledge, this is the first case of poxvirus infection in Canadian squirrels outside of Ontario.

Four free-ranging mink, Neovison vison, collected between June and September 2004 in the Fakahatchee Strand Preserve State Park (FSPSP, Florida, USA), were examined for canine distemper virus (CDV) infection. Microscopic lesions and viral inclusions consistent with CDV infection were observed in three mink. Virus isolation and reverse transcription-polymerase chain reaction performed on all mink were positive for CDV. Anecdotal records of mink observations in FSPSP suggest a postepizootic decline in the mink population followed by an apparent recovery. We recommend further research to assess the status of the Everglades mink and the impact of CDV on this and other American mink populations in Florida.

Canine distemper virus (CDV) has been reported worldwide in wild carnivores and has been cited as the cause of population declines in some species. Crab-eating foxes (Cerdocyon thous) from El Palmar National Park, Argentina, were observed either dead or exhibiting clinical neurologic signs. Samples were taken from two individuals that were later found to be positivefor CDV by direct immunofluorescence in brain tissue. Based on molecular studies, the CDV strain had a highpercentage of identity compared to CDV strains affecting dogs in Argentina.Thisis the first report of CDV infection in wild carnivores in Argentina.

Surveillance for evidence of West Nile virus (WNV) infection in small- and medium-sized wild mammals was conducted in Iowa, USA, from May 2005 to June 2007. Sera were collected from 325 mammals belonging to nine species and tested for antibodies to WNV and other flaviviruses by epitope-blocking enzyme-linked immunosorbent assay (ELISA). All sera that had antibodies to flaviviruses by blocking ELISA were further examined by plaque reduction neutralization test (PRNT). Thirteen mammals were seropositive for WNV by PRNT, including 10 raccoons (Procyon lotor). The seroprevalence for WNV in raccoons was 34%. Although a moderately high seroprevalence for WNV has been detected in raccoons in other surveillance studies in the United States, this has not been reported previously in Iowa or most bordering states. Together, these data indicate that raccoons are exposed to WNV at high rates throughout the United States. Two Virginia opossums (Didelphis virginiana) and one fox squirrel (Sciurus niger) were also seropositive for WNV. Nineteen mammals had antibodies to an undetermined flavivirus(es). In summary, we provide serologic evidence that raccoons in Iowa are commonly exposed to WNV.

During rabies outbreaks in cattle (paralytic rabies) in Argentina associated with the common vampire bat Desmodus rotundus, rabies was observed in marsh deer (Blastocerus dichotomus), red brocket deer (Mazama americana), capybara (Hydrochoerus hydrochaeris), savanna fox (Cerdocyon thous), and great fruit-eating bat (Artibeus lituratus). Rabies could constitute a threat to the survival of marsh deer in places where they live in small groups, and infection of both great fruit-eating bats and savanna fox represent a risk for humans; both species exhibit aggressiveness and fury when infected.

Identifying the source of infectious disease outbreaks is difficult, especially for pathogens that infect multiple wildlife species. Brucella spp. are among the most problematic zoonotic agents worldwide, and they are notoriously difficult to detect and identify. We genotyped 10 variable number of tandem repeat (VNTR) DNA loci in 56 Brucella abortus isolates from bison (Bos bison), elk (Cervus elaphus), and cattle (Bos taurus) to test the wildlife species most likely to be the origin of recent outbreaks of brucellosis in cattle in the Greater Yellowstone Area. Isolates from cattle and elk were nearly identical but highly divergent from bison isolates. These data suggest elk, not bison, are the reservoir species of origin for these cattle infections. This study illustrates the potential power of VNTR genotyping to assess the origin of disease outbreaks, which are increasing worldwide following habitat fragmentation, climate change, and expansion of human and livestock populations.

In 2001, antibodies to Brucella spp. were detected in the wild boar (Sus scrofa) population of Regional Park in the Piedmont, northwest Italy. This was the first report of swine brucellosis in Italian wildlife, and in response, we conducted a survey on wild boars culled in the park to evaluate the presence and epidemiologic pattern of Brucella. In total, 2,267 serum samples and 1,841 tissue samples were collected and tested from 2001 to 2007. Differences in antibody and infection prevalence among sampling years were statistically significant (χ²=28.2, P<0.0001 and χ²=27.8, P<0.0001, respectively). Serologic and culture results also differed between gender and age groups, and for serology, a positive age trend was observed in both male and female age classes (χ^2(trend)=90.9, P<0.00005 OR: 1, 2.5, 4.9, 7.5; χ^2(trend)=43.1, P<0.00002, OR: 1, 2.5, 2.9, 4.8, respectively).

An emaciated, free-ranging, subadult, male beaver (Castor canadensis) was found dead and was necropsied. Microscopically, the beaver had acute necrotizing hepatitis and splenitis with florid lobulated colonies of extracellular coccobacilli. Intravascular septic emboli were identified in lung, small intestine, and kidney, and discrete ulcers with scattered superficial extracellular accumulation of coccobacilli were noted on tail margins and plantar surfaces of the hind feet. Yersinia pseudotuberculosis was cultured on Columbia blood and MacConkey agar and identified by API 20E. Based on the pathology and acute mortality described in this case, as well as historical reports of Y. pseudotuberculosis related mortality in other beavers, this species could serve as a public health sentinel for localized occurrences of this bacterium.

We report a case of tuberculosis due to infection with Mycobacterium bovis in an elderly male black rhinoceros (Diceros bicornis minor) from the Limpopo Province in South Africa. The animal was euthanized due to very poor condition, old age, and dental attrition. Necropsy examination revealed two small nonencapsulated granulomas (∼40-mm diameter) in the dorsocaudal lobe of the left lung. Sequencing of isolated crude lung tissue PCR product and boiled lung culture samples confirmed that the causative organism was M. bovis. Genotyping revealed limited similarities with M. bovis strains isolated thus far from South African cattle or wildlife. The source of the infection could not be determined. This case illustrates that M. bovis could impact conservation of free-ranging rare and endangered species. Effective diagnostics are urgently needed for different animal species, such as white or black rhinoceroses, to certify with a reasonable degree of certainty that these animals are free of tuberculosis in natural habitats.

Eleven species of wild waterfowl (Anseriformes and Charadriiformes) were found dead in the Hongjian Nur Lake in the Ordos wetland, Inner Mongolia of northern China, in 2007. Pasteurella multocida was isolated from tissue samples of dead and sick birds and identified as P. multocida subsp. multocida, serotype A1, using serologic and molecular techniques. Eight bird species in this outbreak had never been previously reported with P. m. multocida infection. This was also the first report of fowl cholera in wild waterfowl in China.

Knowledge of the environmental constraints on a pathogen is critical to predicting its dynamics and effects on populations. Batrachochytrium dendrobatidis (Bd), an aquatic fungus that has been linked with widespread amphibian declines, is ubiquitous in the Rocky Mountains. As part of assessing the distribution limits of Bd in our study area, we sampled the water column and sediments for Bd zoospores in 30 high-elevation water bodies that lacked amphibians. All water bodies were in areas where Bd has been documented from neighboring, lower-elevation areas. We targeted areas lacking amphibians because existence of Bd independent of amphibians would have both ecologic and management implications. We did not detect Bd, which supports the hypothesis that it does not live independently of amphibians. However, assuming a detection sensitivity of 59.5% (based on sampling of water where amphibians tested positive for Bd), we only had 95% confidence of detecting Bd if it was in ≥16% of our sites. Further investigation into potential abiotic reservoirs is needed, but our results provide a strategic step in determining the distributional and environmental limitations of Bd in our study region.

It is unclear whether developmental trends in total leukocyte (WBC) and differential lymphocyte (PropL) counts in peripheral blood of altricial birds typically mirror the known ontogenetic increase in immunocompetence. We documented the development of leukocyte and lymphocyte numbers in peripheral blood of wild, altricial Welcome Swallow (Hirundo neoxena) nestlings. Nestlings had a mass-overshoot–recession growth profile. Hatchlings' mean WBC (7.94×10⁹ cells/l) and PropL (0.65) were respectively 4× and ∼1.7× the mean adult value. Both variables declined at a steady rate throughout nestling development and were 1.3× the mean adult value at fledging. Hatching WBC values that substantially exceeded those of adults could have reflected the parasite- and pathogen-rich nest environment of this species. The developmental declines in peripheral blood WBC and PropL were not inconsistent with an ontogenetic increase in specific immunocompetence; they are likely to have resulted mainly from an increase in the rate of leukocyte trafficking to vulnerable tissues and organs.

We evaluated the small intestines of 123 gray wolves (Canis lupus) that were collected from Idaho, USA (n=63), and Montana, USA (n=60), between 2006 and 2008 for the tapeworm Echinococcus granulosus. The tapeworm was detected in 39 of 63 wolves (62%) in Idaho, USA, and 38 of 60 wolves (63%) in Montana, USA. The detection of thousands of tapeworms per wolf was a common finding. In Idaho, USA, hydatid cysts, the intermediate form of E. granulosus, were detected in elk (Cervus elaphus), mule deer (Odocoileus hemionus), and a mountain goat (Oreamnos americanus). In Montana, USA, hydatid cysts were detected in elk. To our knowledge, this is the first report of adult E. granulosus in Idaho, USA, or Montana, USA. It is unknown whether the parasite was introduced into Idaho, USA, and southwestern Montana, USA, with the importation of wolves from Alberta, Canada, or British Columbia, Canada, into Yellowstone National Park, Wyoming, USA, and central Idaho, USA, in 1995 and 1996, or whether the parasite has always been present in other carnivore hosts, and wolves became a new definitive host. Based on our results, the parasite is now well established in wolves in these states and is documented in elk, mule deer, and a mountain goat as intermediate hosts.

Two cases of Theileria sp. infection in wild Chinese water deer (Hydropotes inermis argyropus) were diagnosed by microscopic examination of thin blood smears and by molecular typing. Two wild Chinese water deer were referred to the Veterinary Medical Center of Chungbuk National University in South Korea, presumably the result of traffic accidents. Thin blood smears, which were stained with Giemsa, revealed intraerythrocytic parasites; however, the deer had no remarkable findings on physical examination. Polymerase chain reaction and direct sequencing indicated that the parasite was nearly identical to the unidentified Theileria sp. previously found in China. Phylogenetic analysis showed that the Theileria sp. identified in these cases are closest to the group of unidentified Theileria spp. previously found in China and Japan.

Nine Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were each inoculated orally with 250 metacercariae of Fasciola hepatica. Blood and fecal samples were collected at the time of inoculation and at 5, 10, 14, and 20 wk after inoculation. Numbers of fluke eggs in feces, hemoglobin, packed cell volume, and eosinophil values were determined. Five of the bighorn sheep were treated with triclabendazole at 40 mg/kg of body weight 14 wk after inoculation. Based on fecal evaluations, all bighorns developed patent infections. Six weeks after treatment, fluke eggs were not detected in feces from the five treated animals but were present in two of four untreated animals. One untreated bighorn sheep was euthanized 20 wk after inoculation, and 57 adult F. hepatica were recovered from the liver. Results from this experiment indicated that bighorn sheep are efficient hosts for F. hepatica. Triclabendazole at 40 mg/kg of body weight was safe and, based on fluke egg recovery in feces, apparently an effective treatment. To my knowledge, this is the first published report of F. hepatica in bighorn sheep.

This study provides the first data on the helminth fauna of the coyote (Canis latrans) in insular Newfoundland. Sixty-nine coyotes were collected between 2001 and 2003 and examined for helminths. A total of 10 helminth species were recorded: the cestodes Taenia ovis krabbei (9%), Taenia hydatigena (4%), Taenia pisiformis (1%), and Mesocestoides spp. (5%); and the nematodes Toxocara canis (19%), Toxascaris leonina (1%), Crenosoma vulpis (19%), Physaloptera rara (6%), Uncinaria stenocephala (3%), and Angiostrongylus vasorum (1%). No significant differences (P≤0.05) were detected between sexes. Mesocestoides spp., T. canis, and C. vulpis were more prevalent in juveniles than adults. Angiostrongylus vasorum is reported in coyotes for the second time in Newfoundland, Canada.

Postmortem examination of seven wild-caught southern giraffes (Giraffa camelopardalis giraffa) from Namibia demonstrated focal discoloration, biliary thickening, and peribiliary fibrosis affecting mainly the left liver lobe. The giraffes were infected with Monodontella giraffae, previously associated with lethal infections in captive okapis (Okapia johnstoni) and giraffes. Contrary to this, all seven giraffes investigated in the present study were clinically healthy. Based on these findings, it is suggested that the nematode M. giraffae may not be an unusual parasite of the giraffe and that it does not necessarily cause detrimental liver disease.

Raccoon (Procyon lotor) carcasses (n=118) were collected from July through December 2007 throughout eastern Tennessee. Necropsies were performed, and Baylisascaris procyonis was collected from the gastrointestinal tract of infected carcasses. Prevalence rates were determined for the overall sample population, males and females, and adults and juveniles. The sample population had a B. procyonis prevalence of 12.7%. Males and females had a prevalence of 15% and 11%, respectively; prevalence in adults and juvenile was 13% and 12.6%, respectively. There were no significant differences in prevalence rates between the different groups. Baylisascaris procyonis is an ascarid infection of raccoons that can infect humans and over 100 species of other animals. The presence of infection in raccoons, paired with the expansion of human populations in eastern Tennessee, is likely to lead to increased interactions between humans and raccoons and therefore an increased risk of human and domestic animal exposure to B. procyonis.

Two species of Phthiraptera, Bovicola caprae (Ischnocera: Bovicolidae) and Solenopotes binipilosus (Anoplura: Linognathi-dae), and one species of mite, Psoroptes ovis (Acari: Psoroptidae), were recorded on huemul (Hippocamelus bisulcus) in Chile. The sucking louse S. binipilosus and the mite P. ovis are reported for the first time from this host.