RT-QuIC detection of chronic wasting disease prion in platelet samples of white-tailed deer.

BACKGROUND: Chronic wasting disease (CWD) is a prion disease of captive and free-ranging cervids. Currently, a definitive diagnosis of CWD relies on immunohistochemistry detection of PrPSc in the obex and retropharyngeal lymph node (RPLN) of the affected cervids. For high-throughput screening of CWD in wild cervids, RPLN samples are tested by ELISA followed by IHC confirmation of positive results. Recently, real-time quacking-induced conversion (RT-QuIC) has been used to detect CWD positivity in various types of samples. To develop a blood RT-QuIC assay suitable for CWD diagnosis, this study evaluated the assay sensitivity and specificity with and without ASR1-based preanalytical enrichment and NaI as the main ionic component in assay buffer. RESULTS: A total of 23 platelet samples derived from CWD-positive deer (ELISA + /IHC +) and 30 platelet samples from CWD-negative (ELISA-) deer were tested. The diagnostic sensitivity was 43.48% (NaCl), 65.22% (NaI), 60.87% (NaCl-ASR1) or 82.61% (NaI-ASR1). The diagnostic specificity was 96.67% (NaCl), 100% (NaI), 100% (NaCl-ASR1), or 96.67% (NaI-ASR1). The probability of detecting CWD prion in platelet samples derived from CWD-positive deer was 0.924 (95% CRI: 0.714, 0.989) under NaI-ASR1 experimental condition and 0.530 (95% CRI: 0.156, 0.890) under NaCl alone condition. The rate of amyloid formation (RFA) was greatest under the NaI-ASR1 condition at 10-2 (0.01491, 95% CRI: 0.00675, 0.03384) and 10-3 (0.00629, 95% CRI: 0.00283, 0.01410) sample dilution levels. CONCLUSIONS: Incorporation of ASR1-based preanalytical enrichment and NaI as the main ionic component significantly improved the sensitivity of CWD RT-QuIC on deer platelet samples. Blood test by the improved RT-QuIC assay may be used for antemortem and postmortem diagnosis of CWD.

Retrotransposon-Based Blood Meal Analysis of Nymphal Deer Ticks Demonstrates Spatiotemporal Diversity of Borrelia burgdorferi and Babesia microti Reservoirs.

Deer tick-transmitted Borrelia burgdorferisensu stricto (Lyme disease) and Babesia microti (babesiosis) increasingly burden public health across eastern North America. The white-footed mouse is considered the primary host for subadult deer ticks and the most important reservoir host for these and other disease agents. Local transmission is thought to be modulated by less reservoir-competent hosts, such as deer, diverting ticks from feeding on mice. We measured the proportion of mouse-fed or deer-fed host-seeking nymphs from 4 sites during 2 transmission seasons by blood meal remnant analysis using a new retrotransposon-based quantitative PCR (qPCR) assay. We then determined the host that was associated with the infection status of the tick. During the first year, the proportion of mouse-fed ticks ranged from 17% on mainland sites to 100% on an island, while deer-fed ticks ranged from 4% to 24%. The proportion of ticks feeding on mice and deer was greater from island sites than mainland sites (on average, 92% versus 43%). Mouse-fed ticks decreased significantly during year 2 in 3 of 4 sites (most were <20%), while deer-fed ticks increased for all sites (75% at one site). Overall, ticks were more likely to be infected when they had fed on mice (odds ratio [OR] of 2.4 and 1.6 for Borrelia and Babesia, respectively) and were less likely to be infected if they had fed on deer (OR, 0.8 and 0.4). We conclude that host utilization by deer ticks is characterized by significant spatiotemporal diversity, which may confound efficacy tests of interventions targeting reservoir hosts.IMPORTANCE White-footed mice are thought to be the most important reservoir host for the deer tick-transmitted pathogens that cause Lyme disease and human babesiosis because they are the primary host for immature ticks. Transmission would be reduced, however, if ticks feed on deer, which are not capable of infecting ticks with either pathogen. By directly measuring whether ticks had fed on either mice or deer using a new quantitative PCR (qPCR) assay to detect remnants of host DNA leftover from the larval blood meal, we demonstrate that host utilization by ticks varies significantly over time and space and that mice often feed fewer ticks than expected. This finding has implications for our understanding of the ecology of these diseases and for the efficacy of control measures.

Anti-Fatigue Peptides from the Enzymatic Hydrolysates of Cervus elaphus Blood.

Red deer (Cervus elaphus) blood is widely used as a health product. Mixed culture fermentation improves the flavor and bioavailability of deer blood (DB), and both DB and its enzymatic hydrolysates exhibit anti-fatigue activities in vivo. To elucidate the bioactive ingredients, enzymatic hydrolysates were fractioned into different peptide groups using reversed phase resin chromatography, and then evaluated using an exhaustive swimming mice model to assess swimming time and biochemical parameters. The structures of the bioactive peptides were elucidated by high performance liquid chromatography with tandem mass detection. Thirty-one compounds were identified as glutamine or branched-chain amino acids containing short peptides, of which Val-Ala-Asn, Val-Val-Ser-Ala, Leu(Ile)-Leu(Ile)-Val-Thr, Pro-His-Pro-Thr-Thr, Glu-Val-Ala-Phe and Val-Leu(Ile)-Asp-Ala-Phe are new peptides. The fractions containing glutamine or valine short peptides, Ala-Gln, Val-Gln, Val-Val-Ser-Ala, Val-Leu(Ile)-Ser improved exercise endurance by increasing hepatic glycogen (HG) storage. The peptides group containing Leu(Ile)-Leu(Ile), Asp-Gln, Phe- Leu(Ile), Val-Val-Tyr-Pro contributed to decreased muscle lactic acid (MLA)accumulation and to an increase in HG. The anti-fatigue activities of DB hydrolysates were attributed to the synergistic effects of different types of peptides.

In vitro detection of haematogenous prions in white-tailed deer orally dosed with low concentrations of chronic wasting disease.

Infectivity associated with prion disease has been demonstrated in blood throughout the course of disease, yet the ability to detect blood-borne prions by in vitro methods remains challenging. We capitalized on longitudinal pathogenesis studies of chronic wasting disease (CWD) conducted in the native host to examine haematogenous prion load by real-time quaking-induced conversion (RT-QuIC) and protein misfolding cyclic amplification. Our study demonstrated in vitro detection of amyloid seeding activity (prions) in buffy-coat cells harvested from deer orally dosed with low concentrations of CWD positive (+) brain (1 gr and 300 ng) or saliva (300 ng RT-QuIC equivalent). These findings make possible the longitudinal assessment of prion disease and deeper investigation of the role haematogenous prions play in prion pathogenesis.

Haematological and serum biochemical reference values in Chinese water deer (Hydropotes inermis): a preliminary study.

BACKGROUND: A selection of haematological and serum biochemical profile was first presented from the 81 samples of Chinese water deer (Hydropotes inermis). The deer health assessment database was initially established, especially in relation to determining potential effects associated with diseases diagnosis. RESULTS: Blood samples were analyzed for different haematological parameters viz. white blood cells (WBC), red blood cells (RBC), haemoglobin (HGB), packed-cell volume (PCV), platelet count (PLT), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean red blood cells distribution width coefficient of variation (RDW) and different hematological parameters viz. total protein (TP), albumin (ALB), globulin (GLB), albumin to globulin ratio (A/G), total bilirubin (TBIL), alkaline phosphatase (ALP), γ-glutamyl transferase (GGT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), AST/ALT, creatinine, urea (BUN), uric acid, total cholesterol (TC), triglyceride, creatine kinase (CK), lactate dehydrogenase (LDH) and cortisol. The adult females had higher values than adult males in albumin, mean corpuscular volume, packed-cell volume, and hemoglobin content values. The deer from Shanghai had higher urea nitrogen values than those from Zhoushan. CONCLUSION: To our knowledge this is the first report about the haematological and serum biochemical parameters in Chinese water deer. We had initially established a profile of Chinese water deer on haematological and serum biochemical parameters based on 81 samples we had collected. The findings can serve as a primary reference for health monitoring and disease prevention in this species.

Characterization of serotonin following exposure to antibiotics in white-tailed deer.

Developing baseline concentrations of serotonin in healthy white-tailed deer will allow for the development of a biomarker using non-invasive sample tissues in sick animals, for example, non-clinical cases of chronic wasting disease. It will also allow some further insight into whether the use of antibiotics as growth promoters (AGP), such as chlortetracycline, is affecting serotonin concentrations in white-tailed deer. Florfenicol and tulathromycin impacts on serotonin concentration changes were also investigated. An analytical method for the detection and confirmation of serotonin, 5-hydroxytryptamine (5-HT), in white-tailed deer tissues was developed and validated. Serum and urine samples were extracted with acetonitrile. Liquid chromatography separation was attained on a Phenomenex C18 column with a Security Guard ULTRA guard column with gradient elution using a mobile phase of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. This methodology was applied to baseline (control), chlortetracycline (CTC) treated, florfenicol treated and tulathromycin treated white-tailed deer serum and urine samples.

Pharmacokinetics of imidocarb dipropionate in white-tailed deer (Odocoileus virginianus) after single intramuscular administration.

This study was designed to investigate the pharmacokinetics of imidocarb, a carbanilide derivative, in white-tailed deer (Odocoileus virginianus). The pharmacokinetic properties of a single intramuscular (IM) dose of imidocarb were determined in 10 deer. A single IM injection of 3.0 mg/kg imidocarb dipropionate was administered, and blood samples were collected prior to, and up to 48 hr after imidocarb administration. Plasma imidocarb concentrations were determined by high-performance liquid chromatography with ultraviolet detection. The disposition of plasma imidocarb was best characterized by a two-compartment open model. The mean ± SE maximal imidocarb concentration in deer was 880.78 ± 81.12 ng/ml at 38.63 ± 5.30 min postinjection. The distribution phase had a half-life (t1/2α ) of 25.90 ± 10.21 min, and plasma imidocarb concentration declined with a terminal elimination half-life (t1/2ß ) of 464.06 ± 104.08 min (7.73 ± 1.73 hr). Apparent volume of distribution based on the terminal phase (VZ /F) was 9.20 ± 2.70 L/kg, and apparent total body clearance (Cl/F) was 15.97 ± 1.28 ml min-1  kg-1 .

Fascioloides magna in Moose ( Alces alces) From Elk Island National Park, Alberta.

Thirty-seven adult female moose ( Alces alces) from 2 distinct but adjacent populations in Elk Island National Park (EINP), Alberta, Canada (19 in north EINP and 18 in south EINP), were fitted with mortality-sensing VHF radio-collars, and radio signals were acquired daily to ascertain mortality status. At capture, serum, whole blood, and feces were collected; pregnancy was determined; teeth were aged by visual inspection; and a portion of liver was assessed by ultrasound examination. Postmortem examination was conducted on 20 suitable carcasses. Clinical pathological abnormalities, including eosinophilia, polycythemia, elevated levels of liver enzymes in serum, hemoglobin, hematocrit, and red blood cell distribution, and liver damage as seen in ultrasound images occurred only in moose from north EINP. Infected moose had 4.7 ± 4.8 Fascioloides magna flukes per liver (mean ± SD). The proportion of moose pregnant at capture was similar in both populations (74% in north EINP, 61% in south EINP). Proportional mortality was significantly higher in moose from the north (68%) than the south (32%). Fascioloides magna was associated as a cause of death in 7 of 14 (50%) moose in the north where cause of death was determined, while predation ( n = 1), acute toxemic syndrome ( n = 3), dystocia ( n = 1), and roadkill and undetermined causes ( n = 3) were additional causes of mortality. F. magna was associated with poor body condition and was a major cause of mortality in north EINP but not south EINP, despite very similar habitat and proximity, suggesting a significant role for these flukes in affecting health and viability of naturally infected moose populations.

Detection of persistent pestivirus infection in pudú (Pudu puda) in a captive population of artiodactyls in Chile.

BACKGROUND: Bovine Viral Diarrhea Virus (BVDV) is the viral agent causing the most important economic losses in livestock throughout the world. Infection of fetuses before their immunological maturity causes the birth of animals persistently infected with BVDV (PI), which are the main source of infection and maintenance of this pathogen in a herd. There is evidence of susceptibility to infection with BVDV in more than 50 species of the order Artiodactyla, and the ability to establish persistent infection in wild cervid species of South America could represent an important risk in control and eradication programs of BVDV in cattle, and a threat to conservation of these wild species. In this study, a serological and virological study was performed to detect BVDV infection in a captive population of non-bovine artiodactyl species in a Chilean zoo with antecedents of abortions whose pathology suggests an infectious etiology. RESULTS: Detection of neutralizing antibodies against BVDV was performed in 112 artiodactyl animals from a zoo in Chile. Three alpacas (Vicugna pacos), one guanaco (Lama guanicoe) and seven pudús (Pudu puda) resulted seropositive, and the only seronegative pudú was suspected to be persistently infected with BVDV. Then two blood samples nine months apart were analyzed by a viral neutralization test and RT-PCR. Non-cytopathogenic BVDVs were isolated in both samples. A phylogenetic analysis showed that the virus was highly related to BVDV-1b strains circulating among Chilean cattle. CONCLUSIONS: This is the first report of a South American deer persistently infected with Bovine Viral Diarrhea Virus. Further studies are needed to determine the possible role of BVDV as a pathogen in pudús and as a threat to their conservation.

Pharmacokinetics of articaine hydrochloride and its metabolite articainic acid after subcutaneous administration in red deer (Cervus elaphus).

AIM: To develop and validate a simple and sensitive method using liquid chromatography-mass spectrometry (LC-MS) for quantification of articaine, and its major metabolite articainic acid, in plasma of red deer (Cervus elaphus), and to investigate the pharmacokinetics of articaine hydrochloride and articainic acid in red deer following S/C administration of articaine hydrochloride as a complete ring block around the antler pedicle. METHODS: The LC-MS method was validated by determining linearity, sensitivity, recovery, carry-over and repeatability. Articaine hydrochloride (40 mg/mL) was administered S/C to six healthy male red deer, at a dose of 1 mL/cm of pedicle circumference, as a complete ring block around the base of each antler. Blood samples were collected at various times over the following 12 hours. Concentrations in plasma of articaine and articainic acid were quantified using the validated LC-MS method. Pharmacokinetic parameters of articaine and articainic acid were estimated using non-compartmental analysis. RESULTS: Calibration curves were linear for both articaine and articainic acid. The limits of quantifications for articaine and articainic acid were 5 and 10 ng/mL, respectively. Extraction recoveries were >72% for articaine and >68% for articainic acid. After S/C administration as a ring block around the base of each antler, mean maximum concentrations in plasma (Cmax) of articaine were 1,013.9 (SD 510.1) ng/mL, detected at 0.17 (SD 0.00) hours, and the Cmax for articainic acid was 762.6 (SD 95.4) ng/mL at 0.50 (SD 0.00) hours. The elimination half-lives of articaine hydrochloride and articainic acid were 1.12 (SD 0.17) and 0.90 (SD 0.07) hours, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The LC-MS method used for the quantification of articaine and its metabolite articainic acid in the plasma of red deer was simple, accurate and sensitive. Articaine hydrochloride was rapidly absorbed, hydrolysed to its inactive metabolite articainic acid, and eliminated following S/C administration as a ring block in red deer. These favourable pharmacokinetic properties suggest that articaine hydrochloride should be tested for efficacy as a local anaesthetic in red deer for removal of velvet antlers. Further studies to evaluate the safety and residues of articaine hydrochloride and articainic acid are required before articaine can be recommended for use as a local anaesthetic for this purpose.

Seroprevalence and herd-level risk factors for seroprevalence of Leptospira spp. in sheep, beef cattle and deer in New Zealand.

AIMS To determine seroprevalence of Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona in beef cattle, sheep and deer in New Zealand and the association between farm-level risk factors and seroprevalence. METHODS Between June 2009 and July 2010, 20 serum samples per flock or herd were collected from 162 sheep flocks, and 116 beef cattle and 99 deer herds from 238 farms, along with farm data by interview. Samples were tested for antibodies to serovars Hardjo and Pomona by microscopic agglutination testing, with a titre ≥48 being positive. Species-specific associations between herd-level seroprevalence (number of seropositive animals, for each serovar, divided by the number of animals tested) and herd-level risk factors were determined by multivariable logistic regression analysis. Vaccinated animals were excluded from seroprevalence estimates but included in multivariable analyses. RESULTS For sheep (n=3,339), animal-level seroprevalence was 43.6 (95% CI=41.9-45.3)% for serovar Hardjo and 14.1 (95% CI=12.9-15.3)% for serovar Pomona; for beef cattle(n=1,886), it was 45.6 (95% CI=43.3-47.9)% for Hardjo and 19.6 (95% CI=17.9-21.5)% for Pomona; and for deer (n=1,870), it was 26.3 (95% CI=24.3-28.4)% for Hardjo, 8.8 (95% CI=7.6-10.2)% for Pomona. In sheep flocks (n=161), flock-level prevalence for Hardjo varied from 77.9-91.3%, and for Pomona from 40.4-73.9%, when ≥1, ≥2 or ≥3 animals were seropositive. In beef herds (n=95), herd-level prevalence for Hardjo varied from 79.0-90.5%, and for Pomona from 42.1-68.4%. In deer herds (n=93), herd-level prevalence for Hardjo varied from 45.2-59.1%, and for Pomona from 22.6-48.4%. For sheep flocks, herd-level seroprevalence for Hardjo was associated with flock size (OR=1.56) and number of dogs (OR=0.75), and for Pomona, seroprevalence varied with region. For beef cattle, herd-level seroprevalence for Hardjo was associated with herd size (OR=1.4), presence of dams (OR=0.6) and vaccination (OR=2.9), and for Pomona, co-grazing with deer (OR=0.4), vaccination (OR=3.22), presence of dams (OR=0.2) and streams (OR=2.7). For deer herds, seroprevalence for Hardjo or Pomona was associated with herd size (OR=1.6 and 1.8) and varied with region, and for Pomona seroprevalence varied with season (summer vs. winter: OR=4.8). CONCLUSIONS Serovars Hardjo and Pomona were highly prevalent at herd and animal levels, with serovar Hardjo highest in all species. Larger herd size was the common risk factor for seroprevalence in all livestock species.

Detection of seroconversion to bovine herpesvirus 1 related alphaherpesvirus and bovine viral diarrhea virusin Polish free-living deer.

There are several infectious agents of domestic cattle that can also be present in free-living ruminant populations. These include bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) which are the causative agents of infectious bovine rhinotracheitis and bovine viral diarrhea, respectively. The study was conducted on serum samples from 59 red deer, 24 roe deer, and 3 fallow deer (86 in total), originating from two geographically separate areas of Poland. The samples were tested with commercially available ELISA tests for BoHV-1 and BVDV. The overall seroprevalence was 5.8% and 3.5%, respectively. All positive samples originated exclusively from red deer. Because of BoHV-1 ELISA cross reactivity with cervid herpesvirus 1 and 2 (CvHV-1 and -2) the nature of alphaherpesviruses infecting the sampled animals could not be assessed.

The influence of selenium and vitamin E supplementation on cytological assessment of red blood cell line of bone marrow in fallow deer kept in captivity.

Cytological evaluation of bone marrow smears stained by May-Grünwald Giemsa method was performed. The smears came from 20 fallow deer (Dama dama) 3 days old divided into 2 groups each consisting of 10 animals. The experimental group (E) received intramuscularly selenium and vitamin E at a dose of 3.0 ml (tocopherol acetate - 50 mg, sodium selenite - 0.5 mg, solvent - 1 ml) in the 3rd day of age. The control group (C) did not receive any supplementation or placebo. For hematological analyzes blood was collected three times: on 0, 15th and 25th day of the experiment. Serum concentration of selenium and vitamin E was determined using high perfor- mance liquid chromatography and glutathione peroxidase activity (GSH-Px) by kinetic method. On the 15th day after supplementation, a statistically significant increase in the percentage of erythroblastic cell line was observed in bone marrow smears. At that time, the increase in GSH-Px activity in the E group was also observed, reaching the value of 165.3 U/gHb, which was statisti- cally significant. The percentage of proerythroblasts (8.23% in group E and 5.02% in group C) differed significantly between groups at the 25th day after supplementation. This study revealed that supplementation of selenium and vitamin E resulted in an increase in the number of erythro- cytes to an average of 13.5 (˟ 10¹²/l) in the experimental group on 25th day with a significant increase in hemoglobin to 193 g/l in the experimental group.

Quantifying capture stress in free ranging European roe deer (Capreolus capreolus).

BACKGROUND: To understand and reduce the concomitant effects of trapping and handling procedures in wildlife species, it is essential to measure their physiological impact. Here, we examined individual variation in stress levels in non-anesthetized European roe deer (Capreolus capreolus), which were captured in box traps and physically restrained for tagging, biometrics and bio-sampling. In winter 2013, we collected venous blood samples from 28 individuals during 28 capture events and evaluated standard measurements for stress (heart rate, body temperature, neutrophil to lymphocyte ratio, lactate and total cortisol). Additionally, we assessed stress using the immunological tool, Leukocyte Coping Capacity (LCC), a real-time proxy for stress measuring oxygen radical production by leukocytes. Finally, the behavioral response to handling was recorded using a scoring system. RESULTS: LCC and therefore stress levels were negatively influenced by the time animals spent in the box trap with human presence at the capture site prior to handling. In contrast, none of the classical stress measures, including total cortisol, nor the behavioral assessment, were correlated with the stressor tested (time of human presence prior to handling) and thus did not provide a clear depiction regarding the extent of the animals short-term stress response. CONCLUSIONS: Overall our study verifies the LCC as a strong method to quantify short-term stress reactions in wildlife. Moreover, our results clearly show that human presence at the trapping site prior to handling should be kept to an absolute minimum in order to reduce stress levels.

High seroprevalence of Toxoplasma gondii in elk (Cervus canadensis) of the central Appalachians, USA.

Toxoplasma gondii is an important protozoan parasite of mammals that impacts animal health and behavior. Although this parasite has been documented in several cervid species, including red deer (Cervus elaphus) in Europe, little is known about T. gondii impacts on the closely related North American counterpart, the elk (wapiti, Cervus canadensis), which has increased in number and expanded in range during the past century. We assessed seroprevalence of T. gondii antibodies using a modified agglutination test (MAT) (1:25 titer) and blood collected from 142 free-ranging elk in Kentucky, USA, where the species was reintroduced during 1997-2002 after over a century of absence. Eighty of 142 (56.3%) elks were seropositive for T. gondii, but we found no infection or titer differences between sexes (U = 2146, P = 0.128). However, odds of T. gondii infection significantly increased with elk age (ß = 0.429, P = 0.001) by a factor of 1.54 (95% CI 1.19-1.99), and titer increased commensurate with age (JT = 3071, P < 0.001). High prevalence of T. gondii infection in elk of this region may be explained by sympatry with two primary hosts, the bobcat and domestic cat, as well as shed oocysts in the soil of this relatively wet and humid region. We suggest that wildlife agency personnel incorporate warnings about proper elk meat preparation into their hunter education outreach programs and literature to reduce the chances for human infection from consuming contaminated venison.

ANTIBODY RESPONSE TO EPSILON TOXIN OF CLOSTRIDIUM PERFRINGENS IN CAPTIVE RED DEER (CERVUS ELAPHUS) OVER A 13-MONTH PERIOD.

Deer are sensitive to clostridial diseases, and vaccination with clostridial toxoids is the method of choice to prevent these infections in ruminants. The purpose of this study was to evaluate the serologic responses in red deer (Cervus elaphus) over a 13-mo period after vaccination with a multivalent clostridial vaccine, containing an aluminium hydroxide adjuvant. Antibody production to the Clostridium perfringens type D epsilon toxin component of the vaccine was measured using an indirect enzyme-linked immunosorbent assay. Animals from group 1 (9 mo old; n = 6) were naïve and received an initial vaccination with a booster vaccine 4 wk apart and one annual booster. Animals from group 2 (21 mo old; n = 10) had been previously vaccinated 12 mo prior and received a first annual booster at the beginning of this study and a second annual booster 12 mo later. The multivalent clostridial vaccine induced a high antibody response that peaked after each injection and then slowly decreased with time. In group 1, a booster vaccine was required to obtain an initial high humoral response. The annual booster injection induced a strong, rapid, and consistent anamnestic response in both groups. The serologic responses persisted significantly over the baseline value for 9-12 mo in group 1, but more than 12 mo in group 2. It is unknown whether the measured humoral immune responses would have been protective as no challenge studies were performed. Further investigation is needed to determine the protective antibody titers to challenge and how long this immunity might persist after vaccination.

LABORATORY VALIDATION OF A POINT-OF-CARE CARDIAC TROPONIN I ASSAY FOR USE IN WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS).

The VetScan® i-STAT® 1 Handheld Analyzer and cardiac troponin I (cTnI) cartridges (i-STAT cTnI assay) measured greater median cTnI concentration [cTnI] in free-ranging white-tailed deer (Odocoileus virginianus) hand-injected with anesthetic drugs after physical restraint in Clover traps than in those ground-darted with the same drugs. This suggested that Clover trapping induces myocardial damage, bringing the use of this capture method under scrutiny. The purpose of this study was to confirm the validity of the i-STAT cTnI assay in deer before recommending changes in capture methods. Median [cTnI] measured by the i-STAT cTnI assay ([cTnI]i) in heparinized whole blood collected from 52 healthy, reproductively mature, female deer physically restrained in a chute was 0.01 ng/ml (10-90% percentiles: 0.00-0.03 ng/ml; minimum, maximum: 0.00, 0.07 ng/ml); [cTnI]i was 0.00 ng/ml in 42% of the deer. There was no association between [cTnI]i and either clotting or hemolytic index. [cTnI]i was 0.00 ng/ml when deer skeletal muscle homogenate was added to deer blood with [cTnI]i of 0.00 ng/ml, confirming the i-STAT cTnI assay does not detect skeletal muscle troponins. When deer cardiac muscle homogenate was serially diluted with 1) deer blood, 2) deer plasma, and 3) cow blood, [cTnI]i was directly proportional (Y intercept=-0.09, 0.7, and -0.08 ng/ml, respectively; r2≥0.97) to the fraction of homogenate in each sample. Deer cardiac muscle homogenate was diluted with deer blood to produce three samples with low, intermediate, and high [cTnI]i; serial measurements (n=10) performed on each sample yielded coefficients of variation (CVs) of 8, 20, and 11%, respectively. Corresponding CVs when plasma was used as diluent were 13, 9, and 7%, respectively. [cTnI]i increased when plasma with a low [cTnI]i was stored at 20-24°C for 9 days. Three freeze-thaw cycles caused no systematic change in plasma [cTnI]i.

DISPOSITION OF CEFTIOFUR AND ITS ACTIVE METABOLITES IN FALLOW DEER (DAMA DAMA) FOLLOWING SINGLE-DOSE INTRAVENOUS AND INTRAMUSCULAR ADMINISTRATION.

Septicemia and foot infections associated with Fusobacterium necrophorum , Pasturella multocida, and Streptococcus suis in captive fallow deer (Dama dama) are reasonably treated with ceftiofur hydrochloride. This study describes the disposition of ceftiofur after single-dose intravenous and intramuscular administration of 3.65±0.1678 mg/kg in six female adult fallow deer using a nonrandomized crossover design and a 7-day washout period. Serial blood samples were collected for 12 hr postdrug administration. Ceftiofur bioactivity, including its active metabolite desfuroylceftiofur, was quantitated in serum using a microbiologic assay. After i.v. administration, the extrapolated serum drug concentration reported as median (range) was 52.83 (43.32-57.49) µg/ml and elimination half-life was 178.36 (19.75-217.22) min. The volume of distribution at steady-state was 0.171 (0.101-0.229) L/kg and serum clearance was 0.97 (0.48-4.3) ml/min per kg. After i.m. administration, median peak plasma concentration (Cmax) was 14.37 (9.00-32.00) µg/ml at 54.5 (11.00-95.00) min. The median elimination half-life and mean residence time were 128.32 (38.03-242.40) and 203.65 (62.48-347.15) min, respectively. The median absorption time after i.m. administration was 14.77 (-57.74 to 94.79) min. Bioavailability of ceftiofur following i.m. administration was 78.00 (58.00-137.00) percent. Based on this study, a mean i.m. dose of ceftiofur of 3.65±0.1678 mg/kg every 12 hr is recommended for maintaining serum concentrations above MIC90 levels for infections associated with F. necrophorum, P. multocida, and S. suis, in addition to other susceptible infectious bacteria.

Circannual changes in serum concentrations of thyroxine, calcitonin and parathormone in immature and mature red deer females (Cervus elaphus).

The aim of this study was to determine circannual changes in the serum concentrations of thyroxine, calcitonin and parathormone in mature and immature red deer females. Blood samples from 8 hinds were collected monthly for 26 months. Secretions of thyroxine and calcitonin showed circannual rhythms with significantly higher levels in the immature hinds compared to the mature animals (p<0.05). For thyroxine, the concentration was higher in the winter/spring period than in summer/autumn (p<0.05), while for calcitonin the concentration profile was the opposite (p<0.05). The concentration of parathormone was significantly higher in summer/autumn that in the other months of the experiment (p<0.01). These results may indicate that the hormones investigated may be involved in the regulation of seasonal reproductive activity and in processes contributing to entering puberty in red deer females.

Haematological parameters do senesce in the wild: evidence from different populations of a long-lived mammal.

Increasing evidence of senescence has been reported from long-term studies of wild populations. However, most studies have focused on life-history traits like survival, reproduction or body mass, generally from a single intensively monitored population. However, variation in the intensity of senescence across populations, and to a lesser extent between sexes, is still poorly understood. In addition, the pattern of age-specific changes in haematological parameters remains virtually unknown to date for any population of vertebrate living in the wild. Using repeated blood samples collected from known-aged (2-15 years of age) roe deer (Capreolus capreolus) from two populations facing highly different environmental conditions, we filled the gap. In particular, we investigated age-specific changes in haematocrit, albumin and creatinine. We reported clear evidence of senescence in all haematological parameters. Moreover, senescence patterns differed between sexes and populations. The rate of senescence was higher in males than in females for haematocrit with no site difference. On the other hand, the rate of senescence in creatinine was higher at Trois Fontaines than at Chizé with no sex difference. Our findings provide a first demonstration of age-specific declines in haematological parameters in wild populations of large herbivores and show that the process of senescence in vertebrates is not restricted to body mass or fitness components. We also demonstrate that the senescence pattern of haematological parameters is context dependent and varies both between sexes and according to environmental conditions.