Knowledge of baseline cutaneous bacterial microbiota may be useful in interpreting diagnostic cultures from captive sick frogs and as part of quarantine or pretranslocation disease screening. Bacteria may also be an important part of innate immunity against chytridiomycosis, a fungal skin disease caused by Batrachochytrium dendrobatidis (Bd). In February 2009, 92 distinct bacterial isolates from the ventral skin of 64 apparently healthy Leiopelma archeyi and Leiopelma hochstetteri native frogs from the Coromandel and Whareorino regions in New Zealand were identified using molecular techniques. The most-common isolates identified in L. archeyi were Pseudomonas spp. and the most common in L. hochstetteri were Flavobacterium spp. To investigate the possible role of bacteria in innate immunity, a New Zealand strain of Bd (Kaikorai Valley-Lewingii-2008-SDS1) was isolated and used in an in vitro challenge assay to test for inhibition by bacteria. One bacterial isolate, a Flavobacterium sp., inhibited growth of Bd. These results imply that diverse cutaneous bacteria are present and may play a role in the innate defense in Leiopelma against pathogens, including Bd, and are a starting point for further investigation.

Upper respiratory tract disease (URTD) in the gopher tortoise (Gopherus polyphemus) is highly contagious and has been implicated in the reduction of populations throughout the range. With the exception of a few limited studies, the prevalence of URTD in Georgia, USA tortoise populations is poorly known. We found that exposure to Mycoplasma agassizii and Mycoplasma testudineum, associated with URTD, varied geographically among 11 Georgia tortoise populations. The prevalence of antibodies to M. agassizii in individual populations was either very low (0–3%, n = 7 populations) or very high (96–100%, n = 4 populations), whereas there was variation in the prevalence of antibodies to M. testudineum among populations (20–61%, n = 10) with only one site being negative. Five sites had tortoises with antibodies to both pathogens, and these were the only sites where we observed tortoises with clinical signs consistent with URTD. We did not find tortoises with clinical signs of URTD at sites with tortoises with antibodies only to M. testudineum, which provides evidence that this organism may be of limited pathogenicity for gopher tortoises. Collectively, these data indicate that both M. agassizii and M. testudineum are present in Georgia populations of gopher tortoises and that clinical disease is apparent in populations where both pathogens are present. Additional research is needed to better understand the role of these two pathogens, and other potential pathogens, in the overall health of tortoise populations, especially if future conservation efforts involve translocation of tortoises.

From 2011–12, we studied a gopher tortoise (Gopherus polyphemus) population with a historically high prevalence of antibodies to Mycoplasma agassizii to assess long-term effects of upper-respiratory-tract disease (URTD) on tortoise behavior. We radiotracked 30 adult tortoises (16 males, 14 females) from a long-term study site with the use of mark–recapture methods to determine site fidelity and to compare home-range size to that of a study in 1997. An additional 10 tortoises (six males, four females) with severe clinical signs of URTD from elsewhere in the study area were radiotracked and compared to tortoises that were asymptomatic or had only mild clinical signs. We also monitored thermoregulatory behavior of tortoises with the use of data loggers affixed to the carapace. There was no significant difference in home-range size between the asymptomatic tortoises and those with mild symptoms. Home ranges of tortoises with severe URTD were significantly larger than asymptomatic or mildly affected tortoises. Tortoises with severe clinical signs moved long distances over short periods, contradicting a hypothesis that chronically infected tortoises are less likely to emigrate. Prevalence of M. agassizii antibodies was similar among the three groups (98% overall), but prevalence of antibodies to a second pathogen associated with URTD, Mycoplasma testudineum, was lower in the asymptomatic (n = 14, 7%) and mild-symptoms (n = 7, 14%) groups than the severe-symptoms group (n = 8, 50%). Variation in the average carapacial temperatures of tortoises with severe URTD was significantly different from carapacial temperatures of mild and asymptomatic tortoises, suggesting differences in thermoregulatory behavior of severely ill tortoises. Our 15-yr recapture data suggest that, despite high prevalence of M. agassizii, population density has not decreased over time. However, emigration, especially of tortoises with severe clinical disease, may play an important role in dispersal and persistence of pathogens.

Mycoplasmas are an important cause of upper respiratory tract disease (URTD) in desert tortoises (Gopherus agassizii) and have been a main focus in attempts to mitigate disease-based population declines. Infection risk can vary with an animal's population of origin, making screening tests popular tools for determining infection status in individuals and populations. To provide additional methods for investigating URTD we developed quantitative PCR (qPCR) assays specific for agents causing clinical signs of URTD: Mycoplasma agassizii, Mycoplasma testudineum, and Testudinid herpesvirus 2 (TeHV2) and tested necropsied desert tortoises housed at the Desert Tortoise Conservation Center in Las Vegas, Nevada, USA, as well as wild desert tortoises (n = 3), during 2010. Findings were compared with M. agassizii enzyme-linked immunosorbent assay (ELISA) data. Based on qPCR, the prevalence of M. agassizii was 75% (33/44) and the prevalence of TeHV2 was 48% (20/42) in the evaluated population. Both agents were also present in the wild tortoises. Mycoplasma testudineum was not detected. The M. agassizii ELISA and qPCR results did not always agree. More tortoises were positive for M. agassizii by nasal mucosa testing than by nasal flush. Our findings suggest that mycoplasmas are not the only agents of concern and that a single M. agassizii ELISA or nasal flush qPCR alone failed to identify all potentially infected animals in a population. Caution should be exercised in using these tests for disposition decisions.

We surveyed pathogens and ectoparasites among federally endangered Amargosa voles (Microtus californicus scirpensis) and sympatric rodents in Tecopa Hot Springs, Inyo County, California, December 2011–November 2012. We aimed to assess disease and detect possible spillover from or connectivity with other hosts within and outside the Amargosa ecosystem. We assessed 71 individual voles and 38 individual sympatric rodents for current infection with seven vector-borne zoonotic pathogens and past exposure to five pathogens. Thirteen percent of Amargosa voles were PCR positive for Toxoplasma gondii, a zoonotic protozoan that may alter host behavior or cause mortality. Additionally, we found antibodies against Borrelia burgdorferi sensu lato (SL) spp. in 21% of voles, against Anaplasma phagocytophilum in 2.6%, Rickettsia spp. in 13%, relapsing fever Borrelia (3.9%), and T. gondii (7.9%). Sympatric rodents also had active infections with Borrelia SL spp. (15%). Of the ectoparasites collected, the tick Ixodes minor is of particular interest because the study area is well outside of the species' reported range and because I. minor ticks infest migratory birds as well as rodents, showing a potential mechanism for pathogens to be imported from outside the Amargosa ecosystem.

Baylisascaris procyonis is a common roundworm of raccoons that causes severe clinical disease in many vertebrates, including humans. The distribution of B. procyonis in the US is poorly documented in portions of its range and has not been reported from Wyoming. Our objectives were to determine the statewide distribution and prevalence of this parasite in raccoons in Wyoming, using intestinal and fecal examinations. We examined 363 raccoons from 23 Wyoming counties in 2009–11, testing the reliability of two methods (intestinal extrusion and incision) to determine worm burdens. We found 163 raccoons (45%) positive for B. procyonis. The two methods of examination did not differ, although extrusion missed some infections. Neither age nor sex affected apparent prevalence or worm burdens. Prevalence did not differ with land use, yet burden was highest among rural raccoons. Fecal examination revealed that juvenile raccoons had a higher proportion of patent infections than adults, but neither sex nor location were indicators of prevalence. Egg density (eggs per gram of feces) did not differ by sex or age; however, rural raccoons had higher egg densities than urban/suburban animals. Understanding the distribution and prevalence of B. procyonis in Wyoming, especially in and around highly populated areas, is an important step in educating the general public and medical community on the potential risks of raccoon roundworm infection.

We assessed the impact of land use, demographic factors, and season on the prevalence and intensity of Baylisascaris procyonis infections in raccoons (Procyon lotor) in Ontario, Canada. From March to October 2012, we recorded the number of B. procyonis in the intestinal tracts of raccoons submitted to the Canadian Cooperative Wildlife Health Centre for necropsy. Logistic regression models were used to examine associations between the presence of B. procyonis and age (adult, juvenile), sex, land use (suburban/urban, rural), and season (March–June and July–October); negative binomial regression models were used to examine associations between the number of worms and the same variables. We detected B. procyonis in 38% (95% confidence interval 30–47%) of raccoons examined (n = 128). In univariable models, the presence of B. procyonis was significantly associated with age, land use, and season (P<0.05). Age was not retained in the multivariable model, and the impact of sex on the presence of B. procyonis varied with land use and season. For example, from March to June, suburban/urban male raccoons were significantly more likely to be infected with B. procyonis than suburban/urban female raccoons. However, later in the summer (July–October), the opposite was true. The median number of worms in the intestinal tracts of infected raccoons was 3 (range 1–116). Worm number was significantly associated with age and season in univariable models; in the multivariable model, juvenile raccoons had significantly more worms than adults, and the impact of season on the number of worms varied with land use and sex. A better understanding of the epidemiology of B. procyonis in raccoons is important for developing appropriate strategies to reduce the risk of human exposure to B. procyonis from the environment.

Although peridomestic passerine species have been involved in influenza A virus (IAV) outbreaks in poultry, there is little evidence to indicate they serve as reservoirs for these viruses under natural conditions. Recent molecular-based detections of IAV in terrestrial wild birds have challenged this paradigm, and it has been suggested that additional research is warranted to better define the role of these birds as IAV hosts. To address this need, we reviewed the published literature reporting results from IAV surveillance of passerines. We also conducted prospective virologic and serologic surveillance of North American passerines for IAVs. The literature review included 60 publications from 1975–2013 that reported results from 829 species of passerines and other terrestrial birds. In our prospective study during 2010 and 2011, 3,868 serum samples and 900 swab samples were collected and tested from 102 terrestrial wild bird species from Georgia, New Jersey, Delaware, and Minnesota, USA. Antibodies to the nucleoprotein of IAV were detected with a commercial blocking enzyme-linked immunosorbent assay in 4/3,868 serum samples (0.1%); all positive samples were from Minnesota. No virus was detected in 900 swab samples by virus isolation in embryonated chicken eggs or matrix real-time reverse transcriptase PCR. Our results are consistent with historic literature; although passerines and terrestrial wild birds may have a limited role in the epidemiology of IAV when associated with infected domestic poultry or other aberrant hosts, there is no evidence supporting their involvement as natural reservoirs for IAV.

Globally, Avipoxvirus species affect over 230 species of wild birds and can significantly impair survival. During banding of nine grassland songbird species (n = 346 individuals) in southwestern Wisconsin, USA, we noted species with a 2–6% prevalence of pox-like lesions (possible evidence of current infection) and 4–10% missing digits (potential evidence of past infection). These prevalences approach those recorded among island endemic birds (4–9% and 9–20% for the Galapagos and Hawaii, respectively) for which Avipoxvirus species have been implicated as contributing to dramatic population declines. Henslow's Sparrow Ammodramus henslowii (n = 165 individuals) had the highest prevalence of lesions (6.1%) and missing digits (9.7%). Among a subset of 26 Henslow's Sparrows from which blood samples were obtained, none had detectable antibody reactive to fowlpox virus antigen. However, four samples (18%) had antibody to canarypox virus antigen with test sample and negative control ratios (P/N values) ranging from 2.4 to 6.5 (median 4.3). Of four antibody-positive birds, two had lesions recorded (one was also missing a digit), one had digits missing, and one had no signs. Additionally, the birds with lesions or missing digits had higher P/N values than did the antibody-positive bird without missing digits or recorded lesions. This study represents an impetus for considering the impacts and dynamics of disease caused by Avipoxvirus among North American grassland bird species.

The lowland tapir (Tapirus terrestris) is found in South America and is listed as Vulnerable to Extinction by the International Union for Conservation of Nature, Red List of Threatened Species. Health issues, particularly infectious diseases, are potential threats for the species. Health information from 65 wild tapirs from two Brazilian biomes, Atlantic Forest (AF) and Pantanal (PA), were collected during a long-term study (1996–2012). The study included physic, hematologic and biochemical evaluations, microbiologic cultures, urinalysis, and serologic analyses for antibodies against 13 infectious agents (viral and bacterial). The AF and PA tapirs were significantly different for several hematologic and biochemical parameters. Ten bacteria taxa were identified in the AF and 26 in the PA. Antibodies against five viruses were detected: Bluetongue virus, eastern equine encephalitis virus, western equine encephalitis virus, infectious bovine rhinotracheitis virus, and porcine parvovirus. A high prevalence of exposure to Leptospira interrogans (10 serovars: Autumnalis, Bratislava, Canicola, Copenhageni, Grippotyphosa, Hardjo, Hebdomadis, Icterohaemorrhagiae, Pomona, and Pyrogenes) was detected in both the AF and PA sites. A greater diversity of serovars and higher antibody titers were found in the PA. Statistically significant differences between sites were found for L. interrogans, equine encephalitis virus, and porcine parvovirus. Based on physical evaluations, both AF and PA populations were healthy. The differences in the overall health profile of the AF and PA tapir populations appear to be associated with environmental factors and infectious diseases ecology. The extensive datasets on hematology, biochemistry, urinalysis, and microbiology results from this paper can be used as reference values for wild tapirs.

American black bears (Ursus americanus) in Maryland, USA, live in forested areas in close proximity to humans and their domestic pets. From 1999 to 2011, we collected 84 serum samples from 63 black bears (18 males; 45 females) in five Maryland counties and tested them for exposure to infectious, including zoonotic, pathogens. A large portion of the bears had antibody to canine distemper virus and Toxoplasma gondii, many at high titers. Prevalences of antibodies to zoonotic agents such as rabies virus and to infectious agents of carnivores including canine adenovirus and canine parvovirus were lower. Bears also had antibodies to vector-borne pathogens common to bears and humans such as West Nile virus, Borrelia burgdorferi, Rickettsia rickettsii, and Anaplasma phagocytophilum. Antibodies were detected to Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Canicola, Grippotyphosa, and Bratislava. We did not detect antibodies to Brucella canis or Ehrlichia canis. Although this population of Maryland black bears demonstrated exposure to multiple pathogens of concern for humans and domesticated animals, the low levels of clinical disease in this and other free-ranging black bear populations indicate the black bear is likely a spillover host for the majority of pathogens studied. Nevertheless, bear populations living at the human–domestic-wildlife interface with increasing human and domestic animal exposure should continue to be monitored because this population likely serves as a useful sentinel of ecosystem health.

Parasites play a major role in ecosystems, and understanding of host–parasite interactions is important for predicting parasite transmission dynamics and epidemiology. However, there is still a lack of knowledge about the distribution, diversity, and impact of parasites in wildlife, especially from remote areas. Hepatozoon is a genus of apicomplexan parasites that is transmitted by ingestion of infected arthropod vectors. However, alternative modes of transmission have been identified such as trophic transmission. Using the 18S rRNA gene as a marker, we provide an assessment of Hepatozoon prevalence in six wild canid and two rodent species collected between 2003 and 2012 from remote areas in North Africa. By combining this with other predator–prey systems in a phylogenetic framework, we investigate Hepatozoon transmission dynamics in distinct host taxa. Prevalence was high overall among host species (African jerboa Jaculus jaculus [17/47, 36%], greater Egyptian jerboa Jaculus orientalis [5/7, 71%], side-striped jackal Canis adustus [1/2, 50%], golden jackal Canis aureus [6/32, 18%], pale fox Vulpes pallida [14/28, 50%], Rüppell's fox Vulpes rueppellii [6/11, 55%], red fox Vulpes vulpes [8/16, 50%], and fennec fox Vulpes zerda [7/11, 42%]). Phylogenetic analysis showed further evidence of occasional transmission of Hepatozoon lineages from prey to canid predators, which seems to occur less frequently than in other predator–prey systems such as between snakes and lizards. Due to the complex nature of the Hepatozoon lifecycle (heteroxenous and vector-borne), future studies on these wild host species need to clarify the dynamics of alternative modes of Hepatozoon transmission and identify reservoir and definitive hosts in natural populations. We also detected putative Babesia spp. (Apicomplexa: Piroplasmida) infections in two canid species from this region, V. pallida (1/28) and V. zerda (1/11).

We immobilized 47 white rhinoceroses (Ceratotherium simum) for dehorning with 1–4 mg of etorphine HCl, 10–40 mg of azaperone, and 7,500 IU of hyaluronidase, at a game ranch in South Africa in November 2012. Forty-four received butorphanol intravenously 5 min after recumbency, at the rate of 10 mg of butorphanol per 1 mg of etorphine, and three animals did not. When possible, blood gas and physiologic parameters were measured immediately before butorphanol administration and 10 min later. Statistically significant improvements were observed, with a reduction in pH, partial pressure of carbon dioxide in arterial blood, heart rate, systolic blood pressure, and diastolic blood pressure, and with an increase in arterial partial pressure of oxygen, arterial hemoglobin oxygen saturation, and respiratory rate in animals administered butorphanol. In the three animals that did not receive butorphanol, no improvement was apparent. Butorphanol given to recumbent white rhinoceroses immediately after immobilization was associated with improved blood gas values and cardiopulmonary function for at least 10 min. Studies on the sustainability of these effects are necessary.

We anesthetized 301 bridled nailtail wallabies (Onychogalea fraenata), captured within Scotia Sanctuary, New South Wales, Australia over four nights in October 2009 to perform health assessments before their release into a predator-proof exclosure. We tested two anesthetic dose-rate combinations, protocol 1 (0.08 mg/kg medetomidine–4.5 mg/kg ketamine), and protocol 2 (0.1 mg/kg medetomidine–5 mg/kg ketamine), each given intramuscularly. Median time from injection to recumbency for protocol 1 was 10 min (1–27 min) and for protocol 2 was 12 min (2–28) (P = 0.12). Five animals died during the induction with protocol 2; none died with protocol 1 (P = 0.06). Physiologic parameters were recorded during recumbency, with no significant abnormalities with protocol 1. Protocol 1 was an effective, efficient regime to anesthetize large numbers of bridled nailtail wallabies under field conditions.

We evaluated the effectiveness of a ketamine-medetomidine-midazolam drug combination administered intramuscularly by remote injection followed by isoflurane anesthesia in free-living aardvarks (Orycteropus afer). Seven aardvarks weighing 33–45 kg were immobilized to perform surgical implantation of temperature loggers using 3.8 mg/kg ketamine, 0.1 mg/kg medetomidine, and 0.25 mg/kg midazolam. Immobilized aardvarks were transported to a surgical theater and received 0.5–1% isoflurane in oxygen after tracheal intubation. After surgery, medetomidine was antagonized with 0.5 mg/kg atipamezole, and aardvarks were released at the site of capture. We recorded induction and recovery times, clinical and physiologic parameters, and conducted blood gas analyses before and during isoflurane administration. Aardvarks showed initial effects within 3 min and reached lateral recumbency within 7 min after drug administration. Heart rate (50–67 beats/min), respiratory rate (10–15 breaths/min), oxygen hemoglobin saturation (SaO₂; 90–97%), and rectal temperature (34.0–37.5 C) were within acceptable physiologic ranges. Mean arterial blood pressure was initially high (146±12 mmHg), but the hypertension resolved over time. Rectal temperature dropped significantly during anesthesia. Four animals had to be treated to relieve apnea. Blood gas analyses revealed mild to moderate hypercapnia but no hypoxaemia. The ketamine-medetomidine-midazolam combination provided effective immobilization. Combined with a low concentration of isoflurane, it can be used for prolonged surgical procedures in wild aardvarks. However, caution is needed, and monitoring of clinical parameters is required.

We report the clinical course and physiologic and anesthetic data for a case series of 76 free-ranging dromedary camels (Camelus dromedarius) chemically restrained, by remote injection from a helicopter, in the rangelands of Western Australia and South Australia, 2008–11, to attach satellite-tracking collars. Fifty-five camels were successfully anesthetized using medetomidine-ketamine (MK, n = 27) and medetomidine-ketamine-butorphanol (MKB, n = 28); the induction of anesthesia in 21 animals was considered unsuccessful. To produce reliable anesthesia for MK, medetomidine was administered at 0.22 mg/kg (±SD = 0.05) and ketamine at 2.54 mg/kg (±0.56), and for MKB, medetomidine was administered at 0.12 mg/kg (±0.05), ketamine at 2.3 mg/kg (±0.39), and butorphanol at 0.05 mg/kg (±0.02). Median time-to-recumbency for MKB (8.5 min) was 2.5 min shorter than for MK (11 min) (P = 0.13). For MK, the reversal atipamezole was administered at 0.24 mg/kg (±0.10), and for MKB, atipamezole was administered at 0.23 mg/kg (±0.13) and naltrexone at 0.17 mg/kg (±0.16). Median time-to-recovery was 1 min shorter for MK (5 min) than MKB (6 min; P = 0.02). Physiologic parameters during recumbency were not clinically different between the two regimes. Both regimes were suitable to safely anesthetize free-ranging camels; however, further investigation is required to find the safest, most consistent, and logistically practical combination.

We describe the pathologic and immunohistochemical findings in five cases of rabies in the insectivorous Pallas's mastiff bat (Molossus molossus) in the city of Patos, state of Paraíba, northeastern Brazil. All cases were found during the day, in different neighborhoods, prostrate and unable to fly. No significant lesions were observed at necropsy or on histologic examination. The brains of the five bats showed strong immunohistochemical labeling for rabies virus in the form of large corpuscles or multiple clusters of granules within the perikarya of neurons in different areas of the brain, mainly in the cerebral cortex and cerebellar Purkinje cells. Three bats tested by direct immunofluorescence and mouse inoculation were also positive for rabies. These data demonstrate that M. molossus may be infected with the rabies virus and develop rabies, even without histologic lesions in the central nervous system, and can be a source of infection for humans and domestic animals.

Leptin is a hormone that plays a key role in regulating energy intake, appetite, and metabolism. In some mammals, leptin has been shown to circulate at levels proportional to body fat, which could make it useful for nonlethal evaluation of body condition. Leptin's usefulness for estimating fat levels (i.e., body condition) of white-tailed deer (Odocoileus virginianus) is unknown. We quantified serum leptin concentrations in a sample of free-ranging, female deer collected in July 2008 and March 2009 from coastal North Carolina, USA. We compared leptin concentrations with kidney fat index, femur marrow fat index, and kidney fat mass. Additionally, we assessed differences in leptin concentrations between the two seasons, lactating and nonlactating females, and gestating and nongestating females. Leptin concentrations were similar between seasons but were lower in lactating and gestating females. We did not detect significant relationships between leptin and the body fat metrics, indicating that leptin may have limited value for estimating fat reserves in white-tailed deer.

Hemochromatosis (iron storage disease) has been reported in diverse mammals including bottlenose dolphins (Tursiops truncatus). The primary cause of excessive iron storage in humans is hereditary hemochromatosis. Most human hereditary hemochromatosis cases (up to 90%) are caused by a point mutation in the hfe gene, resulting in a C282Y substitution leading to iron accumulation. To evaluate the possibility of a hereditary hemochromatosis-like genetic predisposition in dolphins, we sequenced the bottlenose dolphin hfe gene, using reverse transcriptase–PCR and hfe primers designed from the dolphin genome, from liver of affected and healthy control dolphins. Sample size included two case animals and five control animals. Although isotype diversity was evident, no coding differences were identified in the hfe gene between any of the animals examined. Because our sample size was small, we cannot exclude the possibility that hemochromatosis in dolphins is due to a coding mutation in the hfe gene. Other potential causes of hemochromatosis, including mutations in different genes, diet, primary liver disease, and insulin resistance, should be evaluated.

We examined 31 free-ranging coyotes (Canis latrans) from central Georgia, USA, for select parasites and viral agents. Sixteen coyotes had adult heartworms (Dirofilaria immitis). Serum samples from 27 animals revealed antibodies against canine parvovirus (100%), canine distemper virus (48%), canine adenovirus (37%), and Trypanosoma cruzi (7%); none were detected against Leishmania spp. Twenty-two of 24 (92%) coyotes were positive for Toxoplasma gondii. Real-time PCR of feces revealed 32% of coyotes were shedding canine parvovirus, and sequencing revealed type 2b and 2c. Because coyotes could be a spillover host of domestic dog (Canis lupus familiaris) pathogens, studies of the transmission of pathogens between coyotes and domestic dogs are warranted.

Toxoplasma gondii is an obligate intracellular protozoan parasite and a commonly encountered pathogen in humans and animals. The wild boar (Sus scrofa coreanus) is considered a good indicator when monitoring environmental contamination by T. gondii. We surveyed the prevalence of antibodies against T. gondii in wild boars from South Korea. Blood samples were collected from 426 wild boars captured in eight provinces of South Korea during the hunting seasons in 2008–12. Antibodies against T. gondii were detected using an indirect enzyme-linked immunosorbent assay in samples from 152 of boars, indicating an overall antibody prevalence of 36% (95% confidence interval = 31–40%).

We observed predation by river otters (Lontra canadensis) on large numbers of Florida cooter (Pseudemys floridana) and Florida softshell turtles (Apalone ferox) in two small lakes in North Central Florida, USA during a period of unusually low water levels. Carcasses were strewn on the shoreline and accumulated around floating boat docks, where some residents observed turtles being killed. We found 76 carcasses, including predominantly skeletons, and two live, severely injured turtles from one lake; however, numerous remains undoubtedly were unrecovered. The otters frequently eviscerated the turtles and removed the head and one or more appendages, including the phallus of mature males. In skeletal remains, injuries inflicted by otters were nonspecific, indistinguishable from damage caused by scavengers, or easily missed in incomplete carcasses. This report of large-scale mortality of freshwater turtles in Florida suggests that otters could have a significant impact on local turtle populations.

Wildlife is the most important reservoir of Trichinella spp. worldwide. Although the Balkans are a recognized European endemic region for Trichinella infections, data on wildlife are scarce. To monitor the circulation of these zoonotic parasites in the Central Balkan region, the wolf (Canis lupus) was selected because of its abundance (>2,000 individuals) and because it is at the top of the food chain. A total of 116 carcasses of wolves were collected in Serbia and in the neighboring areas of Macedonia and Bosnia and Herzegovina (B&H) between 2006 and 2013. Trichinella spp. larvae were found in 54 (46.5%) wolves. The great majority (90.5%) originated from Serbia, where 52 of the 105 examined animals were Trichinella positive (49.5%; 95% confidence interval  = 39.9–59.1). One positive animal each was found in B&H and Macedonia. All larvae were identified as Trichinella britovi. The high prevalence of Trichinella infection in wolves suggests that this carnivore can be a good indicator species for the risk assessment of the sylvatic Trichinella cycle in the Central Balkans.

Seasonal dynamics of influenza A viruses (IAVs) are driven by host density and population immunity. Through an analysis of subtypic data for IAVs isolated from Blue-winged Teal (Anas discors), we present evidence for seasonal patterns in the relative abundance of viral subtypes in spring and summer/autumn.

In autumn of 2010 we collected fecal samples from the rectums of 89 trapped or road-killed Pennsylvania raccoons (Procyon lotor). Similar samples were collected in the summer and autumn of 2011 from 383 raccoons. Fecal samples were stored in 10% formalin until examined. Using saturated sugar flotation and a direct smear, we found Baylisascaris procyonis eggs in 38% of 2010 samples and 32.9% of 2011 samples. Prevalence in raccoons was greater in autumn than in summer and greater in juveniles than in adults; there was not a statistically significant difference between sexes. Infected raccoons were found in 54 of the 65 counties from which samples were recovered (a mean of 5.9 [range 1–12] raccoons were examined per county). The prevalences were similar in all regions of the state.

In 2011, the Minnesota Rehabilitation Center submitted four dead Black-capped Chickadees (Poecile atricapillus) for necropsy to the Minnesota Veterinary Diagnostic Laboratory. All four chickadees were underweight and dehydrated and their intestinal contents were watery and yellowish. No significant lesions were observed upon histopathologic examination. Viral particles of the family Reoviridae were detected after negative-contrast electron microscopic examination of intestinal contents. Analysis by reverse transcriptase PCR and sequencing confirmed the presence of a reovirus. Based on phylogenetic analysis, the chickadee reovirus had 97.1% to 98.3% and 89.4% to 97.8% nucleotide identity with turkey enteric reoviruses from apparently healthy and enteritis-affected turkey poults, respectively. The chickadee reovirus had only 56.5% and 58.5% nucleotide and 54.5% and 56.8% amino acid identity with psittacine reovirus and Tvarminne avian virus, respectively. These results indicate that the chickadee reovirus is closely related to turkey reoviruses.

Paralytic shellfish poisoning (PSP) is an acute toxic illness in humans resulting from ingestion of shellfish contaminated with a suite of neurotoxins (saxitoxins) produced by marine dinoflagellates, most commonly in the genus Alexandrium. Poisoning also has been sporadically suspected and, less often, documented in marine wildlife, often in association with an outbreak in humans. Kittlitz's Murrelet (Brachyramphus brevirostris) is a small, rare seabird of the Northern Pacific with a declining population. From 2008 to 2012, as part of a breeding ecology study, multiple Kittlitz's Murrelet nests on Kodiak Island, Alaska, were monitored by remote cameras. During the 2011 and 2012 breeding seasons, nestlings from several sites died during mild weather conditions. Remote camera observations revealed that the nestlings died shortly after consuming sand lance (Ammodytes hexapterus), a fish species known to biomagnify saxitoxin. High levels of saxitoxin were subsequently documented in crop content in 87% of nestling carcasses. Marine bird deaths from PSP may be underreported.

Sporadic and geographically widespread reports of parasites affecting the Belted Kingfisher (Megaceryle alcyon) have been published but few have described details of the pathology. A female, adult kingfisher was found dead in a heavily wooded area of a zoo in Maryland, USA. At necropsy, numerous sexually dimorphic, 4.4–40.5-cm adult Monopetalonema alcedinis nematodes were found tightly wound within the coelomic cavity between organs and completely filling the caudal thoracic and abdominal air sacs. Abundant, 30–60-µm diameter, larvated, thick-walled ova were found in the bronchi and parabronchi, within the mesentery, and in the serosa of multiple coelomic organs. Monopetalonema alcedinis is a characteristic member of the superfamily Diplotriaenoidea, a group of nematodes occurring in birds and reptiles. Infective larvae within an invertebrate intermediate host are ingested and penetrate the intestine, traveling to the lungs and then into the air sacs, where the adult females release eggs. The ova are coughed up by the avian host and passed in feces. Specimens of M. alcedinis have been found in the Belted Kingfisher, although typically intensity of infection is low and infections remain asymptomatic. In contrast, we report the second documented case of high numbers of M. alcedinis resulting in pathologic changes in which parasitism contributed to host mortality.

Clostridium perfringens is an important pathogen causing sudden death syndrome, necrotic enteritis, and gas gangrene in ruminants, especially some deer species. Père David's deer (Elaphurus davidianus) is one of the world's rare species and is an endangered and protected species in China. Some Père David's deer in the Chinese Shishou Père David's Deer Preserve died due to C. perfringens infection. We investigated the toxin types and C. perfringens enterotoxin-positive (cpe ) strains of isolated C. perfringens in Père David's deer in China. We collected 155 fecal samples from the Beijing Nanhaizi Père David's Deer Park and the Jiangsu Dafeng Père David's Deer National Nature Reserve between July 2010 and July 2011. Bacteria isolated using blood agar and mannitol agar plates were identified by Gram staining and nested PCR for 16S rRNA. We isolated C. perfringens from 41 fecal samples and used PCR amplification of five toxin genes to identify the toxinotypes and the cpe strains of C. perfringens. Twenty-one isolates were type A, 15 were type E, and five were type D. Fifteen isolates were cpe strains, including eight that were type A and seven that were type E.

As coyotes (Canis latrans) adapt to living in urban environments, the opportunity for cross-species transmission of pathogens may increase. We investigated the prevalence of antibodies to pathogens that are either zoonotic or affect multiple animal species in urban coyotes in the Denver metropolitan area, Colorado, USA, in 2012. We assayed for antibodies to canine parvovirus-2, canine distemper virus, rabies virus, Toxoplasma gondii, Yersinia pestis, and serotypes of Leptospira interrogans. Overall, 84% of the animals had antibodies to canine parvovirus-2, 44% for canine distemper virus, 20% for T. gondii (IgG), 28% for Y. pestis, and 4% for L. interrogans serotype Grippotyphosa. No neutralizing antibodies were detected to rabies virus, T. gondii (IgM), or L. interrogans serotypes other than Grippotyphosa. With 88% of animals exposed to at least one pathogen, our results suggest that coyotes may serve as important reservoirs and sentinels for etiologic agents.

We formulated novel drug combinations of nalbuphine HCl and medetomidine HCl (NalMed), with or without azaperone tartrate, for use in immobilizing Rocky Mountain elk (Cervus elaphus nelsoni) and potentially for other wildlife species. Using the lowest tested nalbuphine dose (0.3 mg/kg) that produced sedation in elk, we initially evaluated a combination of nalbuphine, medetomidine, and azaperone (NalMed-A) for immobilizing adult elk. Based on initial success, we then conducted follow-up trials to assess alternative NalMed formulations successively modified to improve field usability, striving to shorten induction within a dose volume that accommodated practical remote delivery. All NalMed formulations immobilized adult elk; however, combinations with dose volumes that included about 80 mg nalbuphine tended to yield the shortest inductions (mean 6.8 min with, and 7.7 min without, azaperone). Our findings demonstrate that nalbuphine and medetomidine can be combined to yield effective, low-volume (≤2 mL), lightly regulated, reversible drug combinations. Based on results to date, we recommend NalMed-A (40 mg/mL nalbuphine, 10 mg/mL medetomidine, and 10 mg/mL azaperone) dosed at 1.8–2.0 mL for immobilizing adult elk; for recovery we recommend 50 mg naltrexone and 600 mg tolazoline administered intramuscularly (IM) about 5 min in advance of 100 mg atipamezole (divided 25 mg intravenously and 75 mg IM). Further work is under way to explore efficacy in other large mammal species.

The presence of extended-spectrum β-lactamase– and Klebsiella pneumoniae carbapenemase–producing Enterobacteriaceae was investigated in feces and in choanal and cloacal swabs of birds in rescue centers. Ceftiofur-resistant Escherichia coli and Citrobacter freundii were isolated from feces of birds of prey (12% positive) and a group of gulls. The genes bla_CTX-M-1, bla_CTX-M-15, and bla_CTX-M-32 coded for resistance.

Two anesthesia protocols using ketamine plus medetomidine and reversed with atipamezole were assessed for anesthesia of 148 wood mice (Apodemus sylvaticus) sampled under field conditions in northeastern Spain. In both protocols, induction was rapid (mean 76 sec), but recovery was slow (100 min), and mortality was relatively high (9%).

We examined the role of wild birds in the epidemiology of avian coronaviruses by studying oropharyngeal swabs from 32 wild bird species. The 14 avian coronaviruses detected belonged to the gamma-coronaviruses and shared high nucleotide sequence identity with some previously identified strains in wild waterfowl, but not with infectious bronchitis viruses.

No coronavirus was detected by PCR in lung and intestine samples of 100 bats, mostly common pipistrelles (Pipistrellus pipistrellus), collected dead between 2008 and 2013 for rabies surveillance in Belgium. The negative results contrast with the high prevalence of coronaviruses detected in fecal pellets from live-captured bats in some European countries.

We analyzed 32 specimens from nine species of Mustelidae for Trichinella; six infections from two Trichinella species were observed from three host species. This provides documentation of Trichinella in Mustela erminea and Martes foina in Romania and Trichinella spiralis in a mustelid host from Europe.

Trichinella spiralis continues to be a public challenge characterized by a wide host range and geographical distribution (Pozio 2007). During the past 20 yr, Romania has had the most reported human cases of trichinellosis in the world (Blaga et al. 2007). Transmission occurs among domesticated swine, rats, and wild mammals that feed by scavenging or predation (Pozio 2000). Trichinella transmission to humans may occur by consumption of meat of livestock infected after exposure to wildlife (Pozio et al. 2009).

We describe the isolation of West Nile virus (WNV; Flaviviridae, Flavivirus) from blood of a Virginia opossum (Didelphis virginiana) collected in northwestern Missouri, USA in August 2012. Sequencing determined that the virus was related to lineage 1a WNV02 strains. We discuss the role of wildlife in WNV disease epidemiology.

We opportunistically evaluated a combination of acepromazine maleate and medetomidine HCl for use in sedating Rocky Mountain elk (Cervus elaphus nelsoni) and black bears (Ursus americanus) as an alternative to scheduled drug combinations. This combination was safe and effective with limitations inherent in its sedative rather than anesthetic properties.

Anaplasma bovis is one of the most important tick-borne pathogens in China. In the first report of A. bovis in China, we describe infection in eight of 17 wild Reeves' muntjac (Muntiacus reevesi) from Guangxi, southwest China.

A European Robin (Erithacus rubecula) found dead in England had marked blepharitis and periocular alopecia associated with Aprocta cylindrica (Nematoda: Aproctidae) and concurrent mixed fungal infections. Aprocta cylindrica should be considered a differential diagnosis in periocular abnormalities of robins and other insectivorous, migratory passerines in Western Europe.

Hematology, biochemical analyses, and body condition indices are useful tools for describing animal health, especially when making management decisions for species of conservation concern. We report hematologic, biochemical, and body condition index data for 13 free-ranging timber rattlesnakes (Crotalus horridus) sampled repeatedly over an active season in Indiana, USA.

We used fine needle aspiration cytology (FNAC) to diagnose Sertoli cell-only pattern and hypospermatogenesis in an Iberian red deer (Cervus elaphus hispanicus). Cytologic diagnosis was confirmed by histology and epididymal sperm analysis. We conclude that FNAC can be an important diagnostic tool in testicular diseases of wildlife.

We evaluated the duration of infectivity of two isolates of avian paramyxovirus-1 using a model distilled water system. Both viruses remained infective for extended periods with 1-log₁₀ reduction times ranging from 370 (4 C) to 6 (37 C) days. Minimal effects related to pH (5.4–8.4 at 28 C) were observed.

Poland is officially tuberculosis free and bovine tuberculosis (BTB) cases are rarely found except in bovids. We found BTB in a wild boar (Sus scrofa) in the Bieszczady Mountains, southeastern Poland. Studies suggest possible transmission of infection between free-living European bison (Bison bonasus caucasicus) and wild boar in this area.