Coxiella burnetii serostatus in dromedary camels (Camelus dromedarius) is associated with the presence of C. burnetii DNA in attached ticks in Laikipia County, Kenya.

AIMS: Q fever is a globally distributed, neglected zoonotic disease of conservation and public health importance, caused by the bacterium Coxiella burnetii. Coxiella burnetii normally causes subclinical infections in livestock, but may also cause reproductive pathology and spontaneous abortions in artiodactyl species. One such artiodactyl, the dromedary camel (Camelus dromedarius), is an increasingly important livestock species in semi-arid landscapes. Ticks are naturally infected with C. burnetii worldwide and are frequently found on camels in Kenya. In this study, we assessed the relationship between dromedary camels' C. burnetii serostatus and whether the camels were carrying C. burnetii PCR-positive ticks in Kenya. We hypothesized that there would be a positive association between camel seropositivity and carrying C. burnetii PCR-positive ticks. METHODS AND RESULTS: Blood was collected from camels (N = 233) from three herds, and serum was analysed using commercial ELISA antibody test kits. Ticks were collected (N = 4354), divided into pools of the same species from the same camel (N = 397) and tested for C. burnetii and Coxiella-like endosymbionts. Descriptive statistics were used to summarize seroprevalence by camel demographic and clinical variables. Univariate logistic regression analyses were used to assess relationships between serostatus (outcome) and tick PCR status, camel demographic variables, and camel clinical variables (predictors). Camel C. burnetii seroprevalence was 52%. Across tick pools, the prevalence of C. burnetii was 15% and Coxiella-like endosymbionts was 27%. Camel seropositivity was significantly associated with the presence of a C. burnetii PCR-positive tick pool (OR: 2.58; 95% CI: 1.4-5.1; p = 0.0045), increasing age class, and increasing total solids. CONCLUSIONS: The role of ticks and camels in the epidemiology of Q fever warrants further research to better understand this zoonotic disease that has potential to cause illness and reproductive losses in humans, livestock, and wildlife.

Ranavirus (Frog Virus 3) Infection in Free-Living Three-Toed Box Turtles (Terrapene mexicana triunguis) in Missouri, USA.

Frog virus 3 (FV3) and related ranaviruses are emerging infectious disease threats to ectothermic vertebrate species globally. Although the impact of these viruses on amphibian health is relatively well studied, less is understood about their effects on reptile health. We report two cases of FV3 infection, 11 mo apart, in three-toed box turtles (Terrapene mexicana triunguis) from a wildlife rehabilitation center. Case 1 had upper respiratory signs upon intake but had no clinical signs at the time of euthanasia 1 mo later. Case 2 presented for vehicular trauma, had ulcerative pharyngitis and glossitis, and died overnight. In case 1, we detected FV3 nucleic acid with qPCR in oral swabs, kidney, liver, spleen, and tongue. In case 2, we detected FV3 in an oral swab, an oral plaque, heart, kidney, lung, liver, spleen, and tongue. We also detected FV3 nucleic acid with in situ hybridization for case 2. For both cases, FV3 was isolated in cell culture and identified with DNA sequencing. Histopathologic examination of postmortem tissue from case 1 was unremarkable, whereas acute hemorrhagic pneumonia and splenic necrosis were noted in case 2. The difference in clinical signs between the two cases may have been due to differences in the temporal course of FV3 disease at the time of necropsy. Failure to detect this infection previously in Missouri reptiles may be due to lack of surveillance, although cases may also represent a novel spillover to box turtles in Missouri. Our findings reiterate previous suggestions that the range of FV3 infection may be greater than previously documented and that infection may occur in host species yet to be tested.

REFERENCE VALUES AND COMPARISON OF BLOOD CHEMISTRY AND PLASMA PROTEIN VALUES BETWEEN GOLD STANDARD ANALYZERS AND FOUR POINT-OF-CARE DEVICES IN FREE-RANGING CANVASBACKS (AYTHYA VALISINERIA).

Accurate, timely, and cost-effective blood chemistry analysis is an essential tool for directing emergency treatment, monitoring the health status of captive and free-ranging individuals and flocks, and improving the efficacy of conservation actions. Blood samples were obtained from 52 canvasbacks (Aythya valisineria) that were captured on San Francisco Bay, California, during December 2017 as part of a long-term study. Reference values and clinical agreement were determined for blood chemistry and plasma protein parameters among four commonly used point-of-care devices (VetScan® VS2, i-STAT®, AlphaTRAK®2 glucometer, refractometer) and two gold standard laboratory analyzers (Roche cobas® c501, Helena SPIFE 3000 system). Canvasback reference values were generally within expected ranges for Anatidae species with the exception of higher upper limits for sodium and chloride. Creatine kinase and aspartate transaminase values exceeded a published threshold for diagnosis of capture myopathy even though study birds were captured using low-stress techniques and successfully released. With the exception of higher alkaline phosphatase in hatch-year canvasbacks, no age or sex differences were observed for any analyte in this population that was captured during a nonbreeding period. Analysis of analyzer agreement found raw VetScan aspartate transaminase, calcium, glucose, and uric acid values; corrected VetScan albumin, potassium, sodium, and total protein values; raw i-STAT glucose and potassium values; and corrected i-STAT sodium and chloride values were clinically interchangeable with Roche cobas values. Raw VetScan and i-STAT glucose values were also interchangeable. However, none of the Roche or point-of-care analyzer plasma protein values were in clinical agreement with gold standard electrophoresis values. The findings of this study highlight the need for analyzer- or technique-specific reference values and provide biologists and veterinarians quantitative reference values using currently available analyzers to better assess and respond to the health of individuals and populations.

Sublethal effects of parasitism on ruminants can have cascading consequences for ecosystems.

Parasitic infections are common, but how they shape ecosystem-level processes is understudied. Using a mathematical model and meta-analysis, we explored the potential for helminth parasites to trigger trophic cascades through lethal and sublethal effects imposed on herbivorous ruminant hosts after infection. First, using the model, we linked negative effects of parasitic infection on host survival, fecundity, and feeding rate to host and producer biomass. Our model, parameterized with data from a well-documented producer­caribou­helminth system, reveals that even moderate impacts of parasites on host survival, fecundity, or feeding rate can have cascading effects on ruminant host and producer biomass. Second, using meta-analysis, we investigated the links between helminth infections and traits of free-living ruminant hosts in nature. We found that helminth infections tend to exert negative but sublethal effects on ruminant hosts. Specifically, infection significantly reduces host feeding rates, body mass, and body condition but has weak and highly variable effects on survival and fecundity. Together, these findings suggest that while helminth parasites can trigger trophic cascades through multiple mechanisms, overlooked sublethal effects on nonreproductive traits likely dominate their impacts on ecosystems. In particular, by reducing ruminant herbivory, pervasive helminth infections may contribute to a greener world.

A novel herpesvirus detected in 3 species of chelonians.

Herpesviruses are found in free-living and captive chelonian populations, often in association with morbidity and mortality. To date, all known chelonian herpesviruses fall within the subfamily Alphaherpesvirinae. We detected a novel herpesvirus in 3 species of chelonians: a captive leopard tortoise (Stigmochelys pardalis) in western TX, USA; a steppe tortoise (Testudo [Agrionemys] horsfieldii) found near Fort Irwin, CA, USA; and 2 free-living, three-toed box turtles (Terrapene mexicana triunguis) found in Forest Park, St. Louis, MO. The leopard tortoise was coinfected with the tortoise intranuclear coccidian and had clinical signs of upper respiratory tract disease. The steppe tortoise had mucopurulent nasal discharge and lethargy. One of the three-toed box turtles had no clinical signs; the other was found dead with signs of trauma after being observed with blepharedema, tympanic membrane swelling, cervical edema, and other clinical signs several weeks prior to death. Generally, the branching order of the turtle herpesviruses mirrors the divergence patterns of their hosts, consistent with codivergence. Based on phylogenetic analysis, this novel herpesvirus clusters with a clade of viruses that infect emydid hosts and is likely of box turtle origin. Therefore, we suggest the name terrapene alphaherpesvirus 3 (TerAHV3) for the novel virus. This virus also has the ability to host-jump to tortoises, and previously documented herpesviral morbidity tends to be more common in aberrant hosts. The relationship between clinical signs and infection with TerAHV3 in these animals is unclear, and further investigation is merited.

Hematology and biochemistry of critically endangered radiated tortoises (Astrochelys radiata): Reference intervals in previously confiscated subadults and variability based on common techniques.

Madagascar's radiated tortoises (Astrochelys radiata) are critically endangered, threatened by illegal collection, and confiscated in alarming numbers in recent years. Robust population- and technique-specific hematology and biochemistry reference intervals are valuable yet heretofore missing tools for triage, rehabilitation, and reintroduction of confiscated radiated tortoises. We determined reference intervals in 120 previously confiscated, clinically healthy subadult radiated tortoises living under human care within their native habitat at the Tortoise Conservation Center (TCC). Specific analytes measured were manual packed cell volume, total solids, white blood cell (WBC) count and differentials, and biochemistry analytes using a point of care system. To evaluate the effects of different commonly used techniques on these analytes, we compared results between two venipuncture sites (subcarapacial sinus and brachial vein) and three different WBC quantification methods (Natt and Herrick, Leukopet, and slide estimate). Reference intervals were narrower for most analytes, and sodium and potassium were qualitatively higher in the TCC population compared to previously published values from radiated tortoises housed in North American institutions. Creatine kinase, aspartate aminotransferase, glucose and inorganic phosphorus were all significantly greater in brachial samples than in subcarapacial samples. There was poor agreement and evidence of constant and/or proportional bias between all WBC quantification methods. Differences based on time of sample collection were incidentally found in some analytes. These results highlight the need for considering technique, demographic, and environmental factors in creating and applying reference intervals, and contribute foundational knowledge for improving care of radiated tortoises throughout the confiscation-to-release pathway.

A Global Review of Causes of Morbidity and Mortality in Free-Living Vultures.

Vulture species worldwide play a key role in ecosystems as obligate scavengers, and several populations have had precipitous declines. Research on vulture health is critical to conservation efforts including free-living vultures and captive breeding programs, but is limited to date. In this systematic review, we determined the reported causes of free-living vulture species morbidity and mortality worldwide. The most commonly reported cause of mortality was from toxins (60%), especially lead and pesticides, followed by traumatic injury (49%), including collisions with urban infrastructure and gunshot. Neglected areas of research in free-living vulture health include infectious diseases (16%), endocrine and nutritional disorders (6%), and neoplasia (< 1%). Almost half of the studies included in the review were conducted in either Spain or the USA, with a paucity of studies conducted in South America and sub-Saharan Africa. The highest number of studies was on Griffon (Gyps fulvus) (24%) and Egyptian vultures (Neophron percnopterus) (19%), while half of all vulture species had five or fewer studies. Future investigations on free-living vulture health should focus on neglected areas of research, such as infectious diseases, and areas with gaps in the current literature, such as South America, sub-Saharan Africa, and under-studied vulture species.

Fur Color and Nutritional Status Predict Hair Cortisol Concentrations of Dogs in Nicaragua.

This study examined the relationships between hair cortisol concentrations (HCC) and sex, age, nutritional status (as determined by body condition scores, or BCS), and body mass (geometric mean calculated from morphometric measurements), as well as the potential influence of hair pigmentation (light, dark, or agouti/mixed) on HCC in dogs of the Bosawas Biosphere Reserve, Nicaragua. The dogs examined in this study live in a marginal environment where disease, malnutrition, and mortality rates are high. For fur color, HCC was significantly higher in light fur than in than dark and mixed fur (p < 0.001). In addition, BCS scores were found to have a negative effect on HCC (p < 0.001). Measures of sex and body size exhibited inconclusive effects on HCC, and when compared to adult dogs, juvenile dogs did not exhibit significantly different HCC. Repeated measures of dogs over time reveal a moderate intra-class correlation, suggesting that there are unmeasured sources of individual-level heterogeneity. These findings imply a need to account for fur color in studies of HCC in dogs, and the study suggests an overlooked relationship between cortisol and body condition scores in undernourished dogs in diverse settings.

Infectious Diseases, Livestock, and Climate: A Vicious Cycle?

Ruminant livestock are a significant contributor to global methane emissions. Infectious diseases have the potential to exacerbate these contributions by elevating methane outputs associated with animal production. With the increasing spread of many infectious diseases, the emergence of a vicious climate-livestock-disease cycle is a looming threat.

Mortality in Three-Toed Box Turtles (Terrapene mexicana triunguis) at Two Sites in Missouri.

Once ubiquitous, North American box turtles are experiencing reductions in abundance and range, but the magnitude of these losses is largely unknown. In Missouri, native box turtles (Terrapene mexicana triunguis and Terrapene ornata ornata) are declining across the state due to anthropogenic disturbances such as urbanization, habitat fragmentation, and vehicle collisions. Through radio-tracking over a period of 7 years, we documented the survival of adult three-toed box turtles at two sites in Missouri: Forest Park (urban park) and Tyson Research Center (TRC) (a protected rural forest). Estimated annual survival of adult turtles in Forest Park was 79% (95% CI: 0.68-0.87) while at TRC annual survival was 93% (95% CI: 0.83-0.97). The odds of annual survival for a turtle at TRC were 3.5 times that of a turtle living in Forest Park. "Winter kill," which refers to box turtles found dead on the surface during brumation or within 2 weeks of emergence, was the most frequently documented category of mortality in Forest Park. At TRC, winter kill was not documented; however, the reasons for most deaths were unknown. These data raise questions about the potential of large urban parks as refuges for box turtles, which we may answer by future studies that compare box turtles living in multiple urban and rural settings. Our preliminary data suggest that even the largest urban parks may not be able to sustain populations of box turtles which has severe implications as urbanization continues to degrade and eliminate box turtle habitat throughout their range.

Endemic Skunk amdoparvovirus in free-ranging striped skunks (Mephitis mephitis) in California.

The genus Amdoparvovirus includes the newly discovered skunk amdoparvovirus and the well-characterized Aleutian disease virus which causes significant health impacts in farmed mink worldwide. In 2010-2013, an outbreak of fatal amdoparvovirus-associated disease was documented in free-ranging striped skunks (Mephitis mephitis) from the San Francisco Bay Area of California. To characterize the geographic distribution, earliest occurrence and abundance of this virus, as well as possible impacts on sympatric mustelids of conservation concern, we tested blood samples from skunks throughout California and fishers (Pekania pennanti) from northern California for amdoparvovirus DNA. Amdoparvovirus DNA was detected in 64.8% of sampled skunks (140/216), and test-positive skunks were distributed widely throughout the state, from as far north as Humboldt County and south to San Diego County. The first test-positive skunks were detected from 2004, prior to the 2010-2013 outbreak. No significant spatial or temporal clustering of infection was detected. Although healthy and clinically ill animals tested positive for amdoparvovirus DNA, histopathologic evaluation of a subset from clinically ill skunks indicated that positive PCR results were associated with pneumonia as well as there being more than one inflammatory type lesion. None of 38 fishers were PCR-positive. Given the widespread geographic distribution and lack of a clear epizootic centre, our results suggest the presence of an endemic skunk-associated amdoparvovirus strain or species. However, if the virus is not host-specific, skunks' ubiquitous presence across rural and urban habitats may pose a risk to susceptible domestic and wild species including mustelids of conservation concern such as fishers and Pacific martens (Martes caurina).

Complete genome sequences of the first reported california h16 influenza a viruses.

Two reassortant H16 influenza A viruses were isolated from gulls in California. Seven of the eight segments were most closely related to H16 and H13 isolates from eastern North America and Iceland. Of note is a C-terminal truncation of the nonstructural 1 (NS1) protein in one of the isolates that is usually found in swine H1N1 virus.