Brucellosis is a zoonotic disease with terrestrial or marine wildlife animals as potential reservoirs for the disease in livestock and human populations. The primary aim of this study was to assess the presence of Brucella pinnipedialis in marine mammals living along the Dutch coast and to observe a possible correlation between the presence of B. pinnipedialis and accompanying pathology found in infected animals. The overall prevalence of Brucella spp. antibodies in sera from healthy wild grey seals (Halichoerus grypus; n=11) and harbor seals (Phoca vitulina; n=40), collected between 2007 and 2013 ranged from 25% to 43%. Additionally, tissue samples of harbor seals collected along the Dutch shores between 2009 and 2012, were tested for the presence of Brucella spp. In total, 77% (30/39) seals were found to be positive for Brucella by IS711 real-time PCR in one or more tissue samples, including pulmonary nematodes. Viable Brucella was cultured from 40% (12/30) real-time PCR-positive seals, and was isolated from liver, lung, pulmonary lymph node, pulmonary nematode, or spleen, but not from any PCR-negative seals. Tissue samples from lung and pulmonary lymph nodes were the main source of viable Brucella bacteria. All isolates were typed as B. pinnipedialis by multiple-locus variable number of tandem repeats analysis-16 clustering and matrix-assisted laser desorption ionization-time of flight mass spectrometry, and of sequence type ST25 by multilocus sequence typing analysis. No correlation was observed between Brucella infection and pathology. This report displays the isolation and identification of B. pinnipedialis in marine mammals in the Dutch part of the Atlantic Ocean.

From 2011 to 2017, 4,534 serum samples from 13 wildlife species collected across the US and in one territory (US Virgin Islands) were tested for exposure to Leptospira serovars Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, and Pomona. Of 1,759 canids, 1,043 cervids, 23 small Indian mongooses (Herpestes auropunctatus), 1,704 raccoons (Procyon lotor), and five striped skunks (Mephitis mephitis), 27.0, 44.4, 30.4, 40.8, and 60%, respectively, were antibody positive for any of the six serovars. The most commonly detected serovars across all species were Bratislava and Grippotyphosa. Our results indicate that Leptospira titers are very common in a wide variety of wildlife species. These species may act as important reservoirs in the epidemiological cycle of the pathogen. Additional studies to determine the relationship between serologic evidence and shedding of the pathogen by wildlife are necessary to better understand the risk.

We conducted a 10-yr study to establish whether chronic wasting disease (CWD) was readily transmissible to domestic cattle (Bos taurus) following oral inoculation or by cohousing cattle with captive cervids in outdoor research facilities where CWD was enzootic. Calves (n=12) were challenged orally on one occasion using brain homogenate derived from CWD-infected mule deer (Odocoileus hemionus). Five uninoculated cattle served as unchallenged controls. Two other groups of cattle (n=10–11/group) were housed outdoors for 10 yr in captive cervid research facilities. The environmentally challenged cattle were exposed to CWD-associated prions through common paddocks, feed, and water and via direct daily contact with known and potentially infected mule deer or wapiti (Cervus canadensis) throughout the decade-long study period. None of the exposed cattle developed neurologic disease during the study. We euthanized cattle surviving to 10 yr postchallenge and examined all for lesions or disease-associated prion protein (PrP^d) by histopathology, immunohistochemistry, and western immunoblot analysis of central nervous system and lymphoid tissue. None had evidence of PrP^d accumulation. We conclude that the risks of CWD transmission to cattle following oral inoculation or after prolonged exposure to contaminated environments are low.

Three mite species (Demodex ursi, Ursicoptes americanus, and Sarcoptes scabiei) have been associated with mange in black bears (Ursus americanus). Since the early 1990s, the number and geographic distribution of mange cases in black bears in Pennsylvania, US has increased; however, the causative mites have yet to be completely defined. We evaluated several diagnostic approaches for detection and identification of mites in 72 black bears with severe lesions consistent with mange. Sarcoptes scabiei was morphologically identified in skin scrapes from 66 of the bears; no mites were identified in the remaining six. Histopathologic lesions consistent with sarcoptic mange were observed in 39 of 40 bear skin samples examined, and intralesional mites were observed in samples from 38 of these bears. Samples were collected from a subset of the 72 bears for PCR testing targeting both the internal transcribed spacer (ITS)-2 region and cytochrome c oxidase I (cox1) gene including 69 skin scrapes (ITS-2 only), 56 skin biopsies (ITS-2 and cox1), and 36 fecal samples (ITS-2 and cox1). Skin scrapes were a more sensitive sample for PCR detection than either skin biopsies or fecal samples, and the ITS-2 primers proved more sensitive than cox1. Using a commercial indirect enzyme-linked immunosorbent assay, antibodies to S. scabiei were detected in 45/49 (92%) black bears with confirmed mange and 0/62 (0%) cubs with no gross lesions suggestive of mange and which were born to seronegative sows. Sarcoptes scabiei was the predominant mite associated with mange in black bears in Pennsylvania. Diagnostically, cytologic examination of skin scrapes was the most effective approach for diagnosing active mite infestations in black bears. The evaluated serologic assay accurately detected antibodies to S. scabiei in most bears with confirmed S. scabiei infestations. Additional research is needed to determine the usefulness of this approach for larger scale surveys and for asymptomatic bears.

Host responses to infection with novel pathogens are costly and require trade-offs among physiologic systems. One such pathogen is the fungus Pseudogymnoascus destructans (Pd) that causes white-nose syndrome (WNS) and has led to mass mortality of hibernating bats in eastern North America. Although infection with Pd does not always result in death, we hypothesized that bats that survive infection suffer significant consequences that negatively impact the ability of females to reproduce. To understand the physiologic consequences of surviving infection with Pd, we assessed differences in wing damage, mass-specific resting metabolic rate, and reproductive rate between little brown myotis (Myotis lucifugus) that survived a winter in captivity after inoculation with Pd (WNS survivors) and comparable, uninfected bats. Survivors of WNS had significantly more damaged wing tissue and displayed elevated mass-specific metabolic rates compared with Pd-uninfected bats after emergence from hibernation. The WNS survivors and Pd-uninfected bats did not significantly differ in their reproductive capacity, at least in captivity. However, our metabolic data demonstrated greater energetic costs during spring in WNS survivors compared with uninfected bats, which may have led to other consequences for postpartum fitness. We suggest that, after surviving the energetic constraints of winter, temperate hibernating bats infected with Pd faced a second energetic bottleneck after emerging from hibernation.

We describe the clinicopathologic features of a mortality event characterized by blindness and dermatitis affecting eastern grey kangaroos (Macropus giganteus), secondary to hepatogenous photosensitization. Affected animals exhibited photophobic behavior, blindness, ataxia, recumbency, lethargy, ear shaking, and behavior consistent with distress or depression. The photophobia manifested as abnormal shade-seeking during the day, including finding refuge under or in structures used frequently by people. Severely affected kangaroos were jaundiced and had markedly elevated serum bilirubin and gamma glutamyl-transpeptidase concentrations. Blindness in affected animals was attributed to moderate to severe corneal opacity due to corneal edema and inflammation. Skin lesions were typically subtle on gross examination even in cases which had severe necrotizing dermatitis histologically. Histologic lesions in the liver of affected animals included the presence of acicular clefts typical of steroidal saponins. The outbreak was associated with pasture dominated by the invasive grass, Panicum gilvum, which is a recognized source of saponin-induced photosensitization in livestock.

Wildlife anesthetic protocols must offer rapid inductions and recoveries, be physiologically safe, and be minimally regulated. With this in mind, we evaluated differences in induction and recovery times and physiological parameters in 33 American black bears (Ursus americanus) anesthetized with ketamine-xylazine (KX) or immobilized with a commercial drug combination of butorphanol, azaperone, and medetomidine (BAM). Dose was based on mass estimated from field observations. Bears were housed at Appalachian Bear Rescue, Townsend, Tennessee, US, or free-ranging within the Great Smoky Mountains National Park (Tennessee and North Carolina, US) and chemically immobilized for management purposes. From 11 April to 29 June 2016, we immobilized bears with injection via pole syringe or disposable dart projected from an air-powered dart rifle. Once immobilized, we measured each bear's temperature, respiration (breaths/min), heart rate (beats/min), hemoglobin oxygen saturation (via pulse oximetry), arterial blood gases, and mass (kg). We found no differences in the induction parameters, partial pressures of CO₂, and rectal temperatures. The BAM-treated bears had lower heart and respiratory rates that led to lower hemoglobin oxygen saturation levels (from blood gas analysis, SaO₂). The SaO₂ after treatment with BAM (91.1±0.8%) was lower than with KX (93.4±0.9%). After handling, we reversed KX-treated bears with a x̄=0.2±0.02 mg/kg yohimbine and BAM-treated bears with x̄=1.5±0.1 mg/kg atipamezole and 0.8±0.1 mg/kg naltrexone. We found no differences in the recovery times to increased respiration and to the bear assuming a head-up position. The BAM-treated bears stood and recovered quicker than did KX-treated animals. Based on our observations, BAM appears to offer safe, predictable immobilizations with fewer drawbacks and faster recovery times than KX-treated bears.

We evaluated a test and cull strategy for lowering chronic wasting disease (CWD) prevalence in a naturally-infected, free-ranging mule deer (Odocoileus hemionus) herd wintering in the town of Estes Park, Colorado, US and in nearby Rocky Mountain National Park. We tested 48−68% of the estimated number of adult (≥1 yr old) deer annually for 5 yr via tonsil biopsy immunohistochemistry (IHC), collecting 1,251 samples from >700 individuals and removing IHC-positive deer. Among males, CWD prevalence during the last 3 yr of selective culling was lower (one-sided Fisher's exact test P=0.014) than in the period prior. In contrast, CWD prevalence among females before culling and after culling were equivalent (P=0.777). Relatively higher annual testing of males (mean 77%) compared to females (mean 51%) might have contributed to differences seen in responses to management. A more intensive and sustained effort or modified spatial approach might have reduced prevalence more consistently in both sexes. Limitations of this technique in wider management application include cost and labor as well as property access and animal tolerance to repeated capture. However, elements of this approach could potentially be used to augment harvest-based disease management.

Oral rabies vaccination (ORV) is an effective tactic for wildlife rabies control, particularly for containment of disease spread along epizootic fronts. As part of the continuing evaluation of the ORV program in free-ranging raccoons (Procyon lotor) in the US, 37 raccoons from ORV-baited areas in Pennsylvania were live-trapped and transferred to captivity to evaluate protection against rabies in animals with varying levels of existing neutralizing antibodies, expressed in international units per milliliter (IU/mL). Among the 37 raccoons at the date of capture, 24% (9/37) of raccoons were seronegative (<0.05 IU/mL), 22% (8/37) were low positive (≥0.05–0.11 IU/mL), 27% (10/37) were medium positive (>0.11–<0.5 IU/mL), and 27% (10/37) were high positive (≥0.5 IU/mL). Raccoons were held for 86–199 d between the date of capture and rabies virus challenge. At challenge, 68% (25/37) raccoons were seronegative. The overall survival rate among challenged animals was 46% (17/37). Based on the antibody titers at the time of challenge, survivorship was 24% (6/25) among seronegative animals, 100% (4/4) among low positive animals, 83% (5/6) among medium positive animals, and 100% (2/2) among high positive animals. Evidence of high-titer seroconversion after vaccination is a good surrogate indicator of rabies survival; however, survival rates of approximately 45% (15/35) were found among raccoons with detectable titers below 0.5 IU/mL. In contrast, any detectable titer at the time of challenge (>3 mo after vaccination) appeared to be a surrogate indicator of survival. Overall, we illustrated significant differences in the value of specific titers as surrogates for survival based on the timing of measurement relative to vaccination. However, survivorship was generally greater than 45% among animals with any detectable titer regardless of the timing of measurement. These findings suggest that lower titer cutoffs may represent a valid approach to measuring immunization coverage within ORV management zones, balancing both sensitivity and specificity for estimating herd immunity.

Raptors are a target sentinel species for West Nile virus (WNV) because many are susceptible to WNV disease, they are easily sighted because of their large size, and they often occupy territories near human settlements. Sick and dead raptors accumulate at raptor and wildlife rehabilitation clinics. However, investigations into species selection and specimen type for efficient detection of WNV are lacking. Accordingly, we evaluated dead raptors from north-central Colorado, US and southeast Wyoming, US over a 4-yr period. Nonvascular mature feathers (“quill”), vascular immature feathers (“pulp”), oropharyngeal swabs, cloacal swabs, and kidney samples were collected from raptor carcasses at the Rocky Mountain Raptor Program in Colorado from 2013 through 2016. We tested the samples using real-time reverse transcriptase-PCR. We found that 11% (53/482) of raptor carcasses tested positive for WNV infection. We consistently detected positive specimens during a 12-wk span between the second week of July and the third week of September across all years of the study. We detected WNV RNA most frequently in vascular feather pulp from Cooper's Hawk (Accipiter cooperii). North American avian mortality surveillance for WNV using raptors can obviate necropsies by selecting Cooper's Hawk and Red-tailed Hawk (Buteo jamaicensis) as sentinels and targeting feather pulp as a substrate for viral detection.

Canine distemper virus (CDV) is a highly contagious pathogen that principally infects wildlife and domestic carnivores. Peridomestic species such as raccoons (Procyon lotor) experience outbreaks with high mortality. Clinical signs of infection include anorexia, fever, respiratory infection, and neurologic complications. Although not zoonotic, CDV poses a high risk to unvaccinated domestic animals and the conservation of endangered species. During 2013–16, we opportunistically collected wild and domestic carnivore specimens through a rabies surveillance program in northern Colorado, US. Brainstem and cerebellar tissue samples were independently tested for rabies and CDV by fluorescent antibody test. We tested a total of 478 animals for CDV, comprised of 10 wild and domestic carnivore species. A total of 15% (72/478) of all animals sampled tested positive for CDV, consisting of 24% (71/300) of raccoons and 4% (1/26) of coyotes (Canis latrans), but coinfection with rabies virus was not observed among CDV-positive animals. We extracted RNA from positive tissues, and a reverse-transcription PCR was used to create complementary DNA. We amplified and sequenced the hemagglutinin gene from 60 CDV-positive tissues, and a median joining network and maximum likelihood phylogenetic tree revealed two major lineages among samples. Phylogenetic analysis indicated that our sequences were most similar to the America-2 (n=55) and the America-3 (n=5) CDV lineages circulating in North America. Our results indicated two distinct and distantly related clades of CDV overlapping geographically and temporally among raccoon populations in northern Colorado.

Digital media and digital search tools offer simple and effective means to monitor for pathogens and disease outbreaks in target organisms. Using tools such as Rich Site Summary feeds, and Google News and Google Scholar specific key word searches, international digital media were actively monitored from 2012 to 2016 for pathogens and disease outbreaks in the taxonomic order Lagomorpha, with a specific focus on the European rabbit (Oryctolagus cuniculus). The primary objective was identifying pathogens for assessment as potential new biocontrol agents for Australia's pest populations of the European rabbit. A number of pathogens were detected in digital media reports. Additional benefits arose in the regular provision of case reports and research on myxomatosis and rabbit haemorrhagic disease virus that assisted with current research.

Aeromonas hydrophila is a Gram-negative bacterium ubiquitous to freshwater and brackish aquatic environments that can cause disease in fish, humans, reptiles, and birds. Recent severe outbreaks of disease in commercial channel catfish (Ictalurus punctatus) aquaculture ponds have been associated with a hypervirulent Aeromonas hydrophila strain (VAH) that is genetically distinct from less virulent strains. The epidemiology of this disease has not been determined. Given that research has shown that Great Egrets (Ardea alba) can shed viable hypervirulent A. hydrophila after consuming diseased fish, we hypothesized that Double-crested Cormorants (Phalacrocorax auritus), American White Pelicans (Pelecanus erythrorhynchos), and Wood Storks (Mycteria americana) could also serve as a reservoir for VAH and spread the pathogen during predation of fish in uninfected catfish ponds. All three species, when fed VAH-infected catfish, shed viable VAH in their feces, demonstrating their potential to spread VAH.

The alligator snapping turtle (Macrochelys temminckii) is a freshwater apex predator that has experienced severe population declines throughout its range due to historical overharvesting and habitat degradation. Because of its long lifespan, high trophic level, and limited home range, it is a suitable sentinel species for monitoring environmental contaminants. In Louisiana, US a pilot program aims to augment free-ranging populations by releasing captive-reared individuals. Baseline values of potential environmental contaminants were determined as part of an overall health assessment to evaluate captive-reared alligator snapping turtles for release. Blood samples from 3-yr-old (n=23) and 4-yr-old (n=11) captive-reared alligator snapping turtles were tested for lead (Pb), mercury (Hg), and zinc (Zn) levels by atomic absorption spectrophotometry and cholinesterase (ChE) activity (as a biomarker for organophosphate and carbamate exposure) by the modified Ellman method. Reference intervals were determined for Zn (34 to 295 μg/dL), Hg (0 to 4.8 μg/dL), and ChE (0.17 to 1.65 μmole acetylthiocholine/mL per minute). Elevations of Pb, Zn, or Hg, or decreases in ChE activity levels of this cohort during recapture sampling may indicate point-source intoxications or bioaccumulation, both ultimately attributable to environmental contamination. The released animals may serve as sentinels for biomonitoring of their new habitat for the evaluated toxicants.

Eleven avian paramyxovirus type 6 (APMV-6) isolates from Eurasian Wigeon (n=5; Anas penelope), Mallards (n=2; Anas platyrhynchos), and unknown species of wild ducks (n=4) from Korea were analyzed based on the nucleotide (nt) and deduced amino acid sequences of the fusion (F) gene. Fecal samples were collected in 2010–14. Genotypes were assigned based on phylogenetic analyses. Our results revealed that APMV-6 could be classified into at least two distinct genotypes, G1 and G2. The open reading frame (ORF) of the G1 genotype was 1,668 nt in length, and the putative F0 cleavage site sequence was ¹¹³PAPEPRL¹¹⁹. The G2 genotype viruses included five isolates from Eurasian wigeons and four isolates from unknown waterfowl species, together with two reference APMV-6 strains from the Red-necked Stint (Calidris ruficollis) from Japan and an unknown duck from Italy. There was an N-truncated ORF (1,638 nt), due to an N-terminal truncation of 30 nt in the signal peptide region of the F gene, and the putative F0 cleavage site sequence was ¹⁰³SIREPRL¹⁰⁹. The genetic diversity and ecology of APMV-6 are discussed.

There is considerable temporal and spatial variability in the reproductive rates of Hawaiian monk seals (HMS; Neomonachus schauinslandi). Poor reproductive performance limits the recovery of this endangered species; however, causal factors are not fully understood. There is serologic evidence that HMS are exposed to pathogens that can impact reproductive success, but the prevalence of placental infections in HMS has not been evaluated. Placental tissues (n=50), including tissues from 25% of known HMS births, were opportunistically collected in 2011 from six Northwestern Hawaiian Islands and three main Hawaiian Islands. Reproductive histories of the sampled females were representative of the breeding population, as determined through comparisons in age of primiparity and mature reproductive rate. Placental tissues were examined histologically and screened by PCR for Coxiella burnetii, Brucella spp., Chlamydia spp., Leptospira spp., herpesviruses, and Toxoplasma gondii. There was no histologic evidence of placental pathology, and molecular analyses were negative. These negative results can be used to estimate pathogen prevalence in the nonsampled population. For an approximate population size of 1,300 HMS, we can estimate with 99% confidence that the prevalence of each pathogen tested is 9% or less. This is low relative to other pinnipeds and indicates that factors other than reproductive pathology, such as resource limitation, may drive variability in HMS reproductive rates. Further investigation into the cumulative impacts of resource limitation and other stressors on HMS reproduction is warranted.

From August through December 2015, beachcast bird survey programs reported increased deposition of Common Murres (Uria aalge) on central and northern California beaches, but not on southern California beaches. Coastal wildlife rehabilitation centers received more than 1,000 live, stranded, and debilitated murres from Sonoma County to San Luis Obispo County during August–October. Approximately two-thirds of admitted birds were after-hatch-year birds in emaciated body condition and in various stages of molt, with extremely worn plumage. Necropsies were done on a sample (n=35) of birds to determine the probable cause of death of beachcast carcasses. Most birds examined during necropsy were emaciated, with starvation the most likely cause of death. Birds were also tested for underlying infectious diseases at the US Geological Survey National Wildlife Health Center and harmful algal bloom toxins at the University of California at Santa Cruz and the National Oceanographic and Atmospheric Administration's Northwest Fisheries Science Center. Twenty-four out of 29 tested birds had detectable levels of domoic acid, and no indication of infectious disease was found. Emaciation is thought to be the cause of death for these birds, with a large warm water anomaly and harmful algal bloom playing a secondary detrimental role.

Physiological characteristics, such as blood chemistry values, are valuable for evaluating the health of the animals. To our knowledge, these values have never been reported for the free-ranging Asiatic black bear (Ursus thibetanus; ABB). Thus, 28 blood chemistry values from 50 free-ranging ABBs captured in Jirisan National Park, Republic of Korea, from 2005 to 2016 were evaluated. The aim of this study was to establish blood chemistry reference values for the free-ranging ABBs during both the hibernating and nonhibernating seasons. During hibernation, mean values of creatinine (CRE), total cholesterol, total protein (TP), albumin (ALB), triglycerides, and magnesium were significantly higher than those during nonhibernation; however, mean values of blood urea nitrogen, urea nitrogen to creatinine (U/C) ratio, inorganic phosphorous (IP), aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, lactate dehydrogenase (LDH), high-density lipoprotein cholesterol, and alkaline phosphatase (ALP) were significantly lower. Age differences (young vs. adult) were found in IP, LDH, TP, and ALB values during hibernation and in the U/C ratio, calcium, IP, ALP, creatine kinase myocardial band, CRE, total bilirubin, and uric acid values during nonhibernation. However, there were no sex differences (male vs. female).

Urogenital carcinoma is common in wild California sea lions (Zalophus californianus) along the west coast of the US. From 1979 to 1994, this cancer was observed in 18% (66/370) of necropsied subadult and adult sea lions at The Marine Mammal Center in Sausalito, California. A retrospective review of records from 1 January 2005 to 31 December 2015 was performed to characterize prevalence and characteristics of cancer over this decade. Fourteen percent (263/1917) of necropsied sea lions had cancer, of which 90% (237/263) were urogenital carcinoma. The prevalence of urogenital carcinoma was significantly higher in adults compared to juveniles and subadults. Advanced-stage disease with metastases was identified histologically in 78% (182/232) of cases and was the cause of death in 95% (172/182) of these cases. Metastases were most common in lung and lymph nodes, and hydronephrosis, secondary to ureter obstruction by metastases, was identified in 62% (114/185) of animals with advanced disease. No significant temporal change in prevalence was detected over the decade, and this highly aggressive, fatal cancer remains common in stranded California sea lions.

Blood biochemical and hematology analyses are helpful indicators of the physiologic health of animals, particularly when making conservation and management decisions for threatened species. In this study, we 1) established blood biochemical reference intervals for two populations of threatened, free-ranging ornate box turtles (Terrapene ornata) in northern Illinois during their active season and 2) examined the effects of individual carapace temperature (T_c) on blood biochemical variables by using a Bayesian hierarchical framework. Individual blood variables differed throughout the active season (May–September 2015), but there were few distinct patterns in concentrations over time. When controlling for individual variability, blood biochemical variables potassium, sodium, chloride, ionized calcium, hematocrit (percentage of packed cell volume), and osmolality showed no effect of T_c (i.e., slope estimates for these variables were not credibly different from zero) and had little individual variation. Glucose and urea nitrogen were found to have slopes credibly different from zero, with glucose having an estimated positive slope and urea nitrogen having an estimated negative slope, suggesting different relationships in response to T_c when controlling for individual variability. These physiologic blood data will serve as important baseline reference values for the clinical evaluation of wild ornate box turtles presented for veterinary care or for comparison to other studies of wild populations. Further, this study highlights the importance of considering individual-level effects (e.g., T_c) on physiologic health variables.

There are few studies on pathogens affecting free-ranging pinnipeds from South America. We employed molecular techniques to identify a gammaherpesvirus infection by two putative novel herpesvirus species: Otariid herpesvirus 5 (OtHV-5), possibly associated with ulcerative cutaneous lesions, and Otariid herpesvirus 6 (OtHV-6) in a wild South American fur seal (Arctocephalus australis) that stranded alive in Santa Catarina state, southern Brazil. Here we provide new information regarding pinniped herpesviruses, important for the design of future disease surveillance studies.

Midichloria mitochondrii is a tick-borne intracellular bacterium of the order Rickettsiales, found with high prevalence in the sheep tick (Ixodes ricinus). Midichloria mitochondrii is capable of vertical transmission in the tick, but recently evidence of potential horizontal transmission to the tick hosts through the blood meal has been reported. We investigated the presence of the bacterium in the blood of roe deer (Capreolus capreolus) collected from an area known to be highly infested with I. ricinus ticks. We collected blood and sera samples for 3 yr in Gardouch (Haute Garonne, France) and subjected them to molecular screening through PCR and to serological investigation using enzyme-linked immunosorbent assay and western blot. Bacterial DNA was detected in the blood of four of seven animals, but only at one or two points in time, whereas all sera were positive for M. mitochondrii antigens at all times. Our results indicated that the presence of the bacterium in the blood is transient, but the antibody response appeared to be long-lasting, possibly due to constant exposure to tick bites, and thus to repeated injection of bacteria. The role of M. mitochondrii in the mammalian host, and its interaction with other tick-borne bacteria, remains unknown.

We generated the complete sequence of the fusion (F) protein gene from six pigeon paramyxoviruses type 1 (PPMV-1) isolated from Mourning Collared-doves (Streptopelia decipiens) in Namibia, Africa between 2016 and 2017. All of the isolates had an F gene cleavage site motif of ¹¹²RRQKRF¹¹⁷ characteristic of virulent viruses. A phylogenetic analysis using the full F gene sequence revealed that the viruses belonged to genotype VIa and were epidemiologically related to PPMV-1s from Asia, Europe, and North America.

Hummingbirds are specialized nectarivores and important ecological pollinators that are the focus of conservation efforts as well as scientific investigations of metabolism and flight dynamics. Despite their importance, basic information is lacking about hummingbird blood cells. We aimed to establish reference intervals for total and differential leukocyte counts from healthy free-ranging hummingbirds in northern California. Hummingbirds were captured in four counties in spring and summer of 2012. A drop of blood was used to prepare smears for total white blood cell estimate and 200-cell differential leukocyte counts. Reference Value Advisor was used for descriptive statistics and calculation of reference intervals. Blood smears from 42 Anna's Hummingbirds (Calypte anna) and 33 Black-chinned Hummingbirds (Archilochus alexandri) were included. The only significant differences in leukocyte counts were due to age, and juvenile hummingbirds had significantly higher lymphocyte counts than adult hummingbirds (P<0.0001). These reference intervals provide robust baseline data to evaluate health status and disease in free-ranging hummingbirds.

Mesocestoides spp. have a cosmopolitan distribution with zoonotic potential. Mesocestoides tetrathyridia were found under the pericardial sac, on the surface of the crop, and in the peritoneal cavity of a hunter-harvested Northern Bobwhite (Colinus virginianus) and a Scaled Quail (Callipepla squamata) collected during the 2016–17 quail hunting season in northwest and southern Texas, US respectively. Molecular analysis indicated that the tetrathyridia from the birds likely belonged to an undescribed species and are identical to pretetrathyridium stages recently found in Scincella lateralis skinks in Oklahoma. This is the first report of Mesocestoides from North American birds.

A total of 58 American beavers (Castor canadensis) was immobilized with butorphanol, azaperone, and medetomidine (BAM) for the purpose of health assessments, sex determination, and placement of very high-frequency tail transmitters in a subset of animals. Isoflurane gas anesthesia was available to aid with induction when needed, and all animals received supplementary oxygen. Thirty-one beavers immobilized with a mean (SD) dose of 0.65 (0.15) mg/kg butorphanol, 0.22 (0.05) mg/kg azaperone, and 0.26 (0.06) mg/kg medetomidine did not require supplemental isoflurane during induction and the mean induction time was 8 min (range: 3–21 min). This dose was equivalent to 0.024 (0.005) mL of BAM per kilogram. A total of 29 beavers that were immobilized with a mean (SD) of 0.51 (0.07) mg/kg butorphanol, 0.17 (0.02) mg/kg azaperone, and 0.2 (0.03) mg/kg medetomidine needed supplementary isoflurane at 5% and 5 L/min for <1 min to induce full anesthesia. In none of the beavers did BAM alone provide sufficient depth of anesthesia to drill a hole in the tail for transmitter placement, and supplementary isoflurane was administered to reach a sufficient level of analgesia for the procedure. The beavers were reversed with 5 mg of atipamezole per milligram of medetomidine and 1 mg of naltrexone per milligram of butorphanol. No adverse effects or mortalities were observed. Butorphanol-azaperone-medetomidine can be considered safe for use in American beavers for minor procedures.

Rabies and canine distemper virus infections in wildlife share similar presenting signs. Canine distemper virus was detected using real-time PCR of conjunctival swabs in rabies positive raccoons (22/32) and skunks (7/34) during a concurrent rabies and canine distemper outbreak in Ontario, Canada in 2015–16. Coinfections with both viruses should be considered, particularly in distemper endemic areas that are at risk of rabies incursion.

Multi-drug-resistant Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis were isolated from the tissue samples of a giant panda. The E. coli strain SH-YH-DH that we isolated carried a self-transferable plasmid associated with multi-drug-resistant genes (bla_CTX-M-55, bla_TEM-1, sul1, floR, strB, aac(6′)-Ib, and tetA/R). This strain exhibited phenotypic resistance to gentamicin, cefotaxime, ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, and tetracycline. We report the participation of multi-drug-resistant E. coli in a extraintestinal polyinfection in giant pandas and that further research is warranted to pay attention to the threat of multi-drug-resistant bacteria in the panda population.

Aspergillus fumigatus is one of the most common Aspergillus species causing disease in individual animals but it rarely affects deer species. We report a case of A. fumigatus infection causing mycotic pneumonia and hypertrophic osteopathy in a roe deer (Capreolus capreolus).

We tested the urine and saliva of 137 wild rats (Rattus norvegicus) from Vancouver, Canada, for the presence of Leptospira spp. Only one saliva sample was found positive and two were suspect, all from urine-positive rats (n=81), indicating that active shedding of leptospires in saliva is unlikely to occur.

A free-ranging, male, yearling Guadalupe fur seal (Arctocephalus philippii townsendi) died due to multifocal verminous vasculitis with thrombosis and several embolic infarcts in liver, kidney, and brain. Nematodes extracted from lung blood vessels were identified as Parafilaroides decorus, a parasite normally found in alveoli of California sea lions (Zalophus californianus).

We diagnosed infectious canine hepatitis in a free-ranging brown bear (Ursus arctos horribilis) cub from Alaska, US, found dead in October 2015. Intranuclear inclusion bodies were present in hepatocytes, and immunohistochemistry showed reactivity to adenoviral antigens. Sequencing of the hexon protein of adenovirus showed 100% identity to canine adenovirus 1.

A small number of koalas (Phascolarctos cinereus) presented to wildlife hospitals in Queensland, Australia, with signs of arthritis in one or more joints. Molecular analysis identified Chlamydia pecorum in the tarsal tissue and synovial fluid of an affected joint of a koala, suggesting that in addition to livestock, C. pecorum has the potential to cause arthritis in the koala.

A combination of butorphanol-azaperone-medetomidine was used to immobilize four captive caribou (Rangifer tarandus granti) in Palmer, Alaska, US. The average induction time for this combination was 5:17±2:06 min. Inductions were smooth, and recoveries were excellent. This drug combination may provide an alternative to the use of potent opioids for immobilizing caribou.