Analysis of mitogen-stimulated lymphocyte subset proliferation and nitric oxide production by peripheral blood mononuclear cells of captive elk (Cervus elaphus).

Elk (Cervus claphus) are reservoirs for Brucella abortus, Mycobacterium bovis, and Mycobacterium avium subsp. paratuberculosis, each a serious pathogen of domestic livestock. An understanding of the basic immune responsiveness of elk would aid efforts to develop methods to diagnose and prevent these diseases of elk. Peripheral blood mononuclear cells (PBMC) isolated from captive elk were examined for phenotype, lymphocyte subset proliferative capacity, and ability to produce nitric oxide (NO) upon pokeweed mitogen (PWM) stimulation. Although gamma delta TCR+ cells represented a high percentage of the peripheral blood lymphocyte pool, these cells responded poorly to PWM stimulation. B cells (i.e., sIgM+ cells), conversely, were responsive to PWM stimulation. Addition of PWM to PBMC cultures also resulted in a significant production of nitrite, the stable oxidation product of NO. Similar to other ruminant species, the majority of elk peripheral blood sIgM+ cells co-expressed MHC class II and B-B4, a B cell lineage marker that varies in expression during B cell development. Findings from the present study provide basic information on several parameters of cellular immunity of elk.

Epizootiology of chronic wasting disease in free-ranging cervids in Colorado and Wyoming.

Surveillance and epidemic modeling were used to study chronic wasting disease (CWD), a transmissible spongiform encephalopathy that occurs naturally among sympatric, free-ranging deer (Odocoileus spp.) and Rocky Mountain elk (Cervus elaphus nelsoni) populations in contiguous portions of northeastern Colorado and southeastern Wyoming (USA). We used clinical case submissions to identify endemic areas, then used immunohistochemistry to detect CWD-infected individuals among 5,513 deer and elk sampled via geographically-focused random surveys. Estimated overall prevalence (prevalence, 95% confidence interval) in mule deer (4.9%, 4.1 to 5.7%) was higher than in white-tailed deer (2.1%, 0.5 to 3.4%) or elk (0.5%, 0.001 to 1%) in endemic areas; CWD was not detected in outlying portions of either state. Within species, CWD prevalence varied widely among biologically- or geographically-segregated subpopulations within the 38,137 km2 endemic area but appeared stable over a 3-yr period. The number of clinical CWD cases submitted from an area was a poor predictor of local CWD prevalence, and prevalence was typically > or =1% before clinical cases were first detected in most areas. Under plausible transmission assumptions that mimicked field data, prevalence in epidemic models reached about 1% in 15 to 20 yr and about 15% in 37 to 50 yr. Models forecast population declines once prevalence exceeded about 5%. Both field and model data supported the importance of lateral transmission in CWD dynamics. Based on prevalence, spatial distribution, and modeling, we suggest CWD has been occurring in northeastern Colorado and southeastern Wyoming for >30 yr, and may be best represented as an epizootic with a protracted time-scale.

Safety of Brucella abortus strain RB51 in bull elk.

Some of the elk (Cervus elaphus nelsoni) of the Greater Yellowstone Area (Wyoming, Idaho, Montana; USA) are infected with Brucella abortus, the bacterium that causes bovine brucellosis. Brucella abortus strain RB51 vaccine is being considered as a means to control B. abortus induced abortions in cow elk. However, the most probable vaccination strategies for use in free-ranging elk might also result in some bull elk being inoculated, thus, it is important to insure that the vaccine is safe in these animals. In the winter of 1995, 10 free-ranging bull elk calves were captured, tested for B. abortus antibodies, and intramuscularly inoculated with 1.0 x 10(9) colony forming units (CFU) of B. abortus strain RB51. Blood was collected for hemoculture and serology every 2 wk after inoculation for 14 wk. Beginning 4 mo postinoculation and continuing until 10 mo postinoculation elk were serially euthanized, necropsied, and tissues collected for culture and histopathology. These elk cleared the organism from the blood within 6 wk and from all tissues within 10 mo. No lesions attributable to B. abortus were found grossly and only minimal to mild lymphoplasmacytic epididymitis was found in a few elk on histologic examination. In a separate study, six adult bull elk from Wind Cave National Park (South Dakota, USA) were taken to a ranch near Carrington (North Dakota, USA). Three were orally inoculated with approximately 1.0 x 10(10) CFU of RB51 and three were inoculated with corn syrup and saline. Ninety days post-inoculation semen was examined and cultured from these bulls. Strain RB51 was not cultured from their semen at that time. There were no palpable abnormalities in the genital tract and all elk produced viable sperm. Although they contain small sample sizes, these studies suggest that B. abortus strain RB51 is safe in bull elk.

Safety and efficacy of Brucella abortus strain RB51 vaccine in captive pregnant elk.

Brucella abortus strain RB51 is a laboratory-derived rough mutant of virulent B. abortus strain 2308 used as a vaccine because it induces antibodies that do not react on standard brucellosis serologic tests. Strain RB51 vaccine was evaluated in pregnant captive elk (Cervus elaphus) to determine (1) if it induced abortion and (2) if it protected against abortion following subsequent challenge. The time period of this study (February-June, 1998) was similar to field conditions where elk are vaccinated and possibly exposed to B. abortus. Fourteen elk were randomly and equally divided into vaccinated and control groups. The vaccinated group was vaccinated intramuscularly with 1.03 x 10(10) colony-forming units (CFU) of strain RB51 and seroconverted postvaccination. Antibodies to strain RB51 were detected by a modification of an existing dot-blot assay. Both groups were challenged 40 days postvaccination with 9.8 x 10(6) CFU of B. abortus strain 2308 administered intraconjunctivally. The first abortion occurred 38 days postchallenge. Abortion occurred in all control elk and in five of seven vaccinated elk 5 to 12 wk postchallenge (P = 0.23). Mixed strain RB51 and 2308 infections were present in fetuses and vaginas from the vaccinated group whereas only strain 2308 was cultured from control group fetuses and vaginal swabs. Further evaluation of strain RB51 will be necessary to determine if it will be safe and efficacious in free-ranging pregnant elk.

Influence of hypothalamic-pituitary-adrenocortical hormones on reproductive hormones in gray wolves (Canis lupus).

The release of hypothalamic-pituitary-adrenocortical hormones was studied in intact and neutered gray wolves (Canis lupus) to determine how these hormones interact and affect reproductive hormones. Experiments were performed on adult wolves anesthetized with 400 mg ketamine and 50 mg promazine. Intravenous (i.v.) injections with 50 micrograms ovine corticotropin releasing factor (oCRF) significantly increased adrenocorticotropin (ACTH; P < or = 0.01), cortisol (CORT; P < or = 0.004), and progesterone (P < or = 0.036), but not beta-endorphin (P > or = 0.52). Since neutered wolves demonstrated dose-dependent elevations in response to ACTH, it was concluded that the progesterone was secreted from the adrenal gland. Basal luteinizing hormone (LH) concentrations in neutered wolves were similar before and 60 min after i.v. injection of 1, 5, or 25 IU ACTH (P > or = 0.36) or 2.2 mg/kg cortisol (P = 0.42). Neither 25 IU ACTH (P = 0.55) nor 0.22 mg/kg dexamethasone (P = 0.49) altered the LH response to injection of LH releasing hormone in neutered wolves. Chronic administration of 0.22 mg/kg/day dexamethasone for 3 d did not alter baseline LH concentrations (P = 0.75). Injection of 1.0 mg/kg naloxone (NAL), however, increased LH concentrations relative to baseline values in both intact (P = 0.032) and neutered (P = 0.0005) female wolves, but not in intact (P = 0.19) or neutered males (P = 0.07). These results indicated that in gray wolves (1) oCRF stimulated the release of pituitary and adrenal hormones in a fashion similar to that of other mammals; (2) the adrenal cortex was capable of secreting progesterone into the systemic circulation; (3) exogenous glucocorticoids did not alter LH concentrations; and (4) endogenous opioids may modulate LH secretion in female wolves.

Circannual prolactin rhythm in intact dogs housed outdoors.

Prolactin (PRL) and progesterone (P4) values were collected from eight intact (4 female, 4 male) mixed-breed dogs housed outdoors for a two-year span. A circannual component was significant for PRL for each dog (P less than 0.01) and the rhythm was validated for the population (P less than 0.004). Females had two estrus periods a year demonstrated by a P4 frequency of 23.5 +/- 0.47 wk. A 6-month component was statistically significant for P4 for three dogs and of borderline statistical significance for the fourth dog. Circannual acrophases yielded a statistically significant population rhythm (P = 0.012), whereas the 6-month component was of borderline statistical significance (P = 0.056). No time-macroscopic relationship between PRL cycles and P4 cycles was seen. These findings are consistent with similar PRL rhythms in non-domestic canids and emphasize the importance of considering seasonal effects when interpreting PRL data.

Diazepam-induced feeding in captive gray wolves (Canis lupus).

Diazepam doses of 0.2, 0.4, and 0.8 mg/kg induced feeding in sated gray wolves in a dose-dependent manner (p less than 0.001). Neither 0.8 mg/kg of the benzodiazepine antagonist, beta-CCP (p = 0.36), nor 0.8 mg/kg of the benzodiazepine inverse agonist, beta-CCE (p = 0.85), decreased the diazepam-induced hyperphagia. Five of 6 naive wolves (p = 0.003) ate dry dog food within 15.4 +/- 1.9 min of being given 0.4 mg/kg diazepam and freely chose dog food after the single diazepam administration.

Immobilization of gray wolves (Canis lupus) with sufentanil citrate.

Gray wolves (Canis lupus) were immobilized with 0.5 mg/kg xylazine plus 7.5 micrograms/kg of either sufentanil (n = 8), etorphine (n = 8), or carfentanil (n = 2). Drug doses used in this study were selected to provide consistency for comparison and are not recommended doses for effective immobilization of wolves. Induction times were similar among groups (11.9 +/- 1.0 min). Thirty min after induction, wolves were given either 0.5 mg/kg naloxone hydrochloride plus 0.15 mg/kg yohimbine hydrochloride or saline only intravenously. Arousal times for wolves given naloxone and yohimbine (1.2 +/- 0.1 min) were shorter than wolves given saline (35.5 +/- 6.4 min). Respiratory rates were similar among the three drug groups (6.9 +/- 1.0 breaths/min). One animal given sufentanil then saline was found dead 108 min after induction. Presumptive diagnosis was renarcotization and hypothermia. Results indicated that sufentanil is an effective opioid immobilizing agent for gray wolves.

Physiological responses of red foxes (Vulpes vulpes) to surgery.

Radio transmitters were surgically implanted into the abdomens of red foxes (Vulpes vulpes). Blood samples were taken before, immediately after, and 8 hr after surgery and analyzed for hormonal, biochemical, electrolyte and hematologic changes. Samples were taken at the same times from control foxes. Adrenocorticotropin increased after surgery (P less than 0.05), but returned to pre-surgery values after 8 hr. Cortisol increased and remained elevated in the surgery group relative to pre-surgery values or to control values (P less than 0.05); Triiodothyronine and thyroxine both decreased from post-surgery values 8 hr later (P less than 0.05). Creatine kinase, total bilirubin and aspartate aminotransferase increased after 8 hr in both surgery and control groups (P less than 0.05). Carbon dioxide increased under anesthesia in both groups, but returned to initial values after 8 hr (P less than 0.05). The white blood cell count increased after 8 hr only in the surgery group (P less than 0.05). There were no differences between the groups for any value obtained from the initial blood sample. These data indicate that abdominal surgery results in prolonged adrenocortical activity and decreased thyroid hormone levels, but otherwise has minimal systemic effects in red foxes.

Physiological and behavioral responses of gray wolves (Canis lupus) to immobilization with tiletamine and zolazepam.

We conducted a series of experiments to examine the efficacy of Telazol (TEL) for immobilization of captive gray wolves (Canis lupus). Ten wolves were immobilized with either 5 or 10 mg/kg TEL. There was no difference in induction time (6.5 +/- 0.8 versus 5.8 +/- 1.2 min; P = 0.63) between the two doses, but the time to initial arousal was longer for the higher dose (P = 0.0008). Wolves were again immobilized with 10 mg/kg TEL and upon initial arousal were given additional doses of either 5.0 mg/kg TEL or 2.5 mg/kg ketamine (KET) to maintain immobilization. Wolves given boosters of TEL had longer second recovery times than wolves given KET (P = 0.01). There were no differences in induction times or arousal times for wolves immobilized with TEL that had been reconstituted with sterile water and stored at 20 C for 30 days (P greater than or equal to 0.11) or 60 days (P greater than or equal to 0.27) when compared to immobilization times using fresh solution. Induction times for wolves given TEL reconstituted with water and propylene glycol and stored for 60 days at -9 C were longer (P less than 0.05) than such times for wolves given standard TEL, but time to initial arousal was unchanged (P greater than or equal to 0.44). There were no differences in heart rates (P = 0.36), blood pressures (P = 0.32), respiratory rates (P = 0.91), and rectal temperatures (P = 0.62) between the two TEL doses. Telazol was shown to be an effective and safe immobilizing agent for gray wolves.

Chemical immobilization of red foxes (Vulpes vulpes).

Red foxes (Vulpes vulpes) were immobilized with one of the following drug combinations: ketamine/xylazine (n = 22), ketamine/promazine (n = 35), ketamine/midazolam (n = 13), or tiletamine/zolazepam (n = 22). Foxes given ketamine/xylazine had the shortest induction and longest recovery times relative to other drug combinations, whereas foxes given ketamine/midazolam had the longest induction times. Recommended doses for the various combinations are given. Foxes given ketamine/xylazine were given either 0.1, 0.2, 0.4 mg/kg yohimbine, or saline 40 min after anesthetic induction. Administration of yohimbine significantly shortened arousal and recovery times relative to control values (P less than 0.001).

Physiological response of gray wolves to butorphanol-xylazine immobilization and antagonism by naloxone and yohimbine.

Captive gray wolves (Canis lupus) were immobilized (loss of consciousness) with 2.0 mg/kg xylazine hydrochloride (XYL) and 0.4 mg/kg butorphanol tartrate (BUT) administered intramuscularly. Induction time was 11.8 +/- 0.8 min (mean +/- SE). Immobilization resulted in bradycardia, respiratory depression, and normotension. Fifteen min after induction, six wolves were given either 0.05 mg/kg naloxone hydrochloride (NAL) and 0.125 or 0.250 mg/kg yohimbine hydrochloride (YOH), or an equal volume of saline (control) intravenously. Antagonism resulted in shortened recovery times compared to control animals (P less than 0.03); there was no difference in recovery times between the YOH doses (P greater than 0.05). Antagonism caused increases in heart rate (HR) and respiratory rate (RR), but no changes in MABP. Eight other wolves were similarly immobilized, but given only NAL. This resulted in partial antagonism with the animals appearing to be sedated with XYL only. Three wolves given only 0.4 mg/kg BUT assumed a state described as "apathetic sedation." Three other wolves sedated with only 2.0 mg/kg XYL showed a profound sedation characterized by recumbency, bradycardia and shallow, but regular, respiration. This study demonstrated that (1) BUT and XYL together, but not separately, can completely immobilize wolves, (2) this combination can be rapidly antagonized by NAL and YOH, and (3) there appeared to be no adverse cardiopulmonary reactions to any of the drugs used.

Use of xylazine sedation with yohimbine antagonism in captive gray wolves.

Captive gray wolves (Canis lupus) were given 2.2 mg/kg xylazine hydrochloride intramuscularly resulting in profound sedation in 9.1 +/- 0.6 min (mean +/- SE). Heart rate was 42.0 +/- 1.0 beats per minute and respiratory rate was 20.1 +/- 1.6 respirations per minute during sedation. A variety of manipulations could be performed on sedated animals in relative safety. Thirty min after xylazine administration, the animals were given either 0.15 mg/kg yohimbine hydrochloride or 5% dextrose solution intravenously causing recovery in 5.3 +/- 1.0 and 97.1 +/- 14.0 min, respectively (P less than 0.001).

Xylazine and ketamine-induced glycosuria in white-tailed deer.

This study documents glycosuria effects of xylazine and ketamine in eight captive and 19 free-ranging white-tailed deer (Odocoileus virginianus) from January to April 1985. Mean urinary glucose:creatinine ratios in two groups of deer fed high protein-high energy and low protein-low energy diets and in free-ranging deer were 1,000, 719, and 259, respectively. Glucose did not occur in urine of deer immobilized by physical restraint. Glucose:creatinine increased with the time interval between xylazine injection and urine collection in the two groups of captive deer.

Ketamine-xylazine anesthesia in red-tailed hawks with antagonism by yohimbine.

Five red-tailed hawks (Buteo jamaicensis) were anesthetized at weekly intervals with intravenous ketamine hydrochloride (KET, 4.4 mg/kg) and xylazine hydrochloride (XYL, 2.2 mg/kg). Twenty min after anesthesia, yohimbine hydrochloride (YOH, 0.05, 0.10, 0.20 and 0.40 mg/kg) or a control was administered. All doses of YOH significantly reduced the head-up times (F = 20.84, df = 1,24, P less than 0.0001) and the standing times (F = 12.30, df = 1,24, P less than 0.0001), compared to the control group. The heart and respiratory rates following YOH (all doses) were significantly greater (P less than 0.01) than the anesthetized rates, but were comparable to the rates observed in restrained, unanesthetized hawks. Yohimbine did not appear to have any significant effect on body temperature. Based upon administration of 4.4 mg/kg KET and 2.2 mg/kg XYL, a dose of 0.10 mg/kg YOH was recommended to achieve antagonism without causing profound cardiovascular or respiratory changes.

Immobilization of white-tailed deer by etorphine and xylazine and its antagonism by nalmefene and yohimbine.

White-tailed deer (Odocoileus virginianus) were immobilized with either 4.0 mg etorphine hydrochloride (ETOR) or 3.5 mg ETOR and 50.0 mg xylazine (XYL). Deer immobilized with ETOR only were given 4.0 mg nalmefene hydrochloride (NAL), a new opioid antagonist, 20 min after induction. Deer immobilized with ETOR and XYL received 3.5 mg NAL and 0.125 mg/kg yohimbine hydrochloride (YOH). The dose of 4.0 mg ETOR did not provide acceptable immobilization and was discontinued. A NAL:ETOR ratio of 1:1 was insufficient for complete and sustained antagonism of ETOR. Subsequently, deer were immobilized with ETOR and XYL as before which was then antagonized with 35.0 mg NAL and 0.125 mg/kg YOH. The 10:1 ratio of NAL:ETOR appeared to provide complete antagonism with no evidence of renarcotization. Although more study is required, NAL could become a useful antagonist for opioid-induced immobilizations.

Cardiovascular and behavioral responses of gray wolves to ketamine-xylazine immobilization and antagonism by yohimbine.

Adult wolves (Canis lupus) were immobilized with 6.6 mg/kg ketamine hydrochloride (KET) and 2.2 mg/kg xylazine hydrochloride (XYL) administered intramuscularly. Induction time was 4.6 +/- 0.3 min (mean +/- SE). Immobilization resulted in significant bradycardia and hypertension (P less than 0.05). Twenty min after induction, the wolves were given 0.05-0.60 mg/kg yohimbine hydrochloride (YOH). Yohimbine given intravenously produced dose-related increases in heart rate (HR) with doses greater than 0.15 mg/kg resulting in extreme tachycardia (greater than 300 bpm). All doses of YOH caused a temporary decrease in mean arterial blood pressure (MABP) with some individual animals manifesting profound hypotension (less than 30 torr) at doses greater than 0.15 mg/kg. Increasing the dose of YOH above 0.15 mg/kg did not significantly decrease either arousal or ambulation times. Administering YOH at 40 or 60 min after induction resulted in decreased arousal and ambulation times. Stimulation by weighing and taking repeated blood samples during anesthesia did not shorten arousal times. We recommend that wolves immobilized with XYL-KET be antagonized with doses of YOH less than 0.15 mg/kg.

Xylazine hydrochloride-ketamine hydrochloride immobilization of wolves and its antagonism by tolazoline hydrochloride.

Fourteen wolves (Canis lupus L.) were singularly or repeatedly immobilized with 30 mg xylazine hydrochloride (HCl) and 400 mg ketamine HCl. Mean induction time was 5.3 +/- 4.6 min (mean +/- SD). Administration of 8.0 mg/kg tolazoline HCl as an antagonist significantly reduced immobilization times from 148.0 +/- 52.7 to 47.9 +/- 8.9 min (F = 63.69, df = 1,17, P less than 0.05). The average times from injection to ambulation for 2.0, 4.0, and 8.0 mg/kg tolazoline HCl were 35.2 +/- 31.8, 18.5 +/- 11.7, and 10.2 +/- 9.1 min. Tolazoline HCl increased heart rates significantly (P less than 0.001) from 75 +/- 14 to 120 +/- 23 beats/min, reversing a xylazine HCl-induced bradycardia. Respiratory rates also increased significantly (P less than 0.01) after tolazoline HCl injection from 19 +/- 7 to 28 +/- 8 breaths/min. Immobilization resulted in an initial hypertension which was normalized after tolazoline HCl administration. One female wolf had a single sinoatrial block within 1 min of receiving tolazoline HCl. Tolazoline HCl appears to be an effective antagonist for xylazine HCl-ketamine HCl immobilization of wolves.