The anti-nociceptive and cardiopulmonary effects of extradural fentanyl–xylazine in sheep

Objective To compare the anti‐nociceptive effects of extradural xylazine, fentanyl and a xylazine–fentanyl combination in sheep, and to measure the cardiopulmonary effects of the xylazine–fentanyl combination. Study design Prospective, randomized study. Animals Twenty‐five half‐merino ewes 2–4 years of age and body mass 54.2 ± 1.1 kg. Methods Six sheep in group 1 received 0.2 mg kg^?1 xylazine by extradural injection, six in group 2 received fentanyl 1.5 µg kg^?1 and 13 in group 3 received the combination of both treatments. In all groups, drugs were mixed with saline (0.15 mL kg^?1 before injection). Pulmonary and carotid arterial catheters were placed in seven sheep of group 3 which were used to evaluate cardiopulmonary effects. Anti‐nociception was determined by the response to electrical stimulation (40 V for 1.5 milliseconds) of the left flank and by superficial and deep muscular ‘pinpricking’ stimulation of the pelvic and thoracic limbs and thoracolumbar region. Results Lack of response to electrical stimulation at the left flank was present in 10 ± 1.1 minutes (mean ± SEM) (group 1) and in 4.5 ± 0.5 minutes in group 3. The duration of lack of response to electrical stimulation at the left flank was 96 ± 6 minutes in group 1 and 315 ± 6 minutes in group 3. Responses persisted in group 3. Significant decreases (p < 0.05) in cardiac output 30, 45, 60 and 90 minutes after injection, and in cardiac work at 30 and 45 minutes were observed in the seven animals of group 3. Arterial blood pH was lowest at 90 minutes, arterial bicarbonate was lowest at 60 minutes and values for both arterial and mixed venous base excess increased significantly at 60 and 90 minutes. There was no significant change from baseline values in heart rate, mean arterial blood pressure, respiratory rate, body temperature, systemic vascular resistance, arterial and mixed venous PO₂, PCO₂, oxygen saturation, blood oxygen content, haemoglobin concentration, mixed venous blood bicarbonate and pH. Conclusions Fentanyl decreases the onset time and prolongs the duration of anti‐nociception produced by xylazine. The combination decreases cardiac output but is without significant respiratory effects. Clinical relevance Further studies are required to show that surgery is possible in sheep after extradural xylazine–fentanyl injection.     

Serum pharmacokinetics of oxytetracycline in sheep and calves and tissue residues in sheep following a single intramuscular injection of a long-acting preparation

The pharmacokinetics of a long‐acting oxytetracycline (OTC) formulation (Liquamycin® LA‐200®) injected intramuscularly (i.m.) at a dose of 20 mg/kg were determined in four calves and 24 sheep to determine if the approved label dose for cattle provided a similar serum time/concentration profile in sheep. The AUC for the calves was 168±14.6 (μg ? h/mL) and was significantly less than the AUC for sheep (209±43 μg ? h/mL). Using the standard two‐stage approach and a one‐compartment model, the mean Cmax for the calves was 5.2±0.8 μg/mL, and for the sheep was 6.1±1.3 μg/mL. The mean terminal phase rate constants were 0.031 and 0.033 h, and the Vdss were 3.3 and 3.08 L/kg for the calves and sheep respectively. Analysis of the data using the standard two‐stage approach, the naive pooled‐data approach and a population model gave very similar results for both the cattle and sheep data. Sheep tissue residues of OTC in serum, liver, kidney, fat, muscle and injection site were measured at 1, 2, 3, 5, 7 and 14 days after a single i.m. injection of 20 mg/kg OTC. Half‐lives of OTC residues in the tissues were 38.6, 33.4, 28.6, 25.4, 21.3, and 19.9 h for injection site, kidney, muscle, liver, mesenteric fat and renal fat, respectively. The ratio of tissue to serum concentration was fairly consistent at all slaughter times, except for the fat and injection sites. The mean ratios were 1.72, 4.19, 0.11, 0.061, 0.84 and 827 for the liver, kidney, renal fat, mesenteric fat, muscle and injection sites, respectively. The tissue concentrations of OTC residues were below the established cattle tolerances for OTC in liver (6 p.p.m.), muscle (2 p.p.m.) and kidney (12 p.p.m.) by 48 h, and in injection site muscle by 14 days after the single i.m. injection of 20 mg/kg.     

Effects of central administration of glucagon on feed intake and endocrine responses in sheep

This study was conducted to investigate effects of glucagon intracerebroventricularly administered on feed intake and endocrine changes in sheep. Four male sheep (48–55 kg BW) were used. The animals were acclimatized to be fed alfalfa hay cubes at 12.00 hour. Human glucagon (40 and 80 µg/0.5 mL) was injected into the lateral ventricle at 12.00 hour. Blood samples were taken every 10 min from 30 min before to 180 min after the glucagon injection. Soon after the injection, the animals were given alfalfa hay cubes, and the amounts of the feed eaten within 2 h were measured. Feed intakes were significantly (P < 0.05) suppressed by 80 µg of glucagon. Plasma glucose levels in control animals were gradually decreased after the feeding, whilst those in glucagon‐treated animals were temporarily elevated just after the feeding and then kept higher than control levels. Plasma insulin was abruptly elevated after the feeding and was maintained at higher levels than before the feeding in all treatments. Plasma NEFA concentrations were decreased after the feeding in all treatments. A tendency of increase in plasma cortisol levels occurred in glucagon‐injected animals. The present study provides the first evidence that glucagon directly acts on the brain, then inhibiting feeding behavior and inducing endocrine responses in ruminants.     

The effect of high ambient temperature on Ca,P and Mg balance and bone turnover in high‐yielding dairy cows

We investigated the effect of heat stress on Ca, P and Mg balance and bone turnover in lactating cows. In a 2 × 2 crossover design, four multiparous lactating Holstein cows were kept in a chamber and subjected to a constant moderate (18°C) ambient temperature (MT) or high (28°C) ambient temperature (HT). The cows were fed total mixed ration (Ca, 0.7%; P, 0.4%; Mg, 0.2%) ad libitum. The milk yield under HT (35.4 kg/day) tended to be lower (P < 0.10) than that under MT (43.2 kg/day). The concentrations of milk P (P < 0.05) and Mg (P < 0.01) were significantly lower under HT than MT. The Ca, P and Mg intake (P < 0.10); Ca (P < 0.10), P, and Mg (P < 0.05) secretion into milk; and Ca (P < 0.05), P (P < 0.01), and Mg (P < 0.05) absorption in the intestine were lower under HT than MT. The plasma osteocalcin, a marker of bone turnover, was significantly lower (P < 0.05) under HT than MT. Heat stress did not affect plasma C‐telopeptide of collagen type I, a bone resorption marker, and plasma parathyroid hormone concentration.     

Antinociceptive activity of midazolam in sheep

The purpose of this study was to examine the effects of midazolam on the nociceptive threshold responses in sheep. The intravenous administration of midazolam (0. 1–0.3 mg/kg) produced a significant dose-dependent elevation of the mechanical and thermal nociceptive thresholds. The intravenous administration of flumazenil (20 μg/kg) markedly attenuated the antinociceptive activity of midazolam in the mechanical nociceptive test, whereas intravenous naloxone (0.2 mg/kg) had no significant effect on midazolam-mediated analgesia. The intrathecal administration of midazolam (1 mg), via chronically implanted cervical subarachnoid catheters, produced a significant elevation in the mechanical threshold responses. These results indicate that midazolam has antinociceptive actions in the sheep and suggest that this effect is, at least partially, mediated at the spinal level.     

Kisspeptin‐10 stimulates the release of luteinizing hormone and testosterone in pre‐ and post‐pubertal male goats

The aims of the present study were to clarify the effect of kisspeptin‐10 (Kp10) on the secretion of luteinizing hormone (LH) and testosterone (T) in pre‐pubertal and post‐pubertal male ruminants. Four male goats (Shiba goats) were given an intravenous (i.v.) injection of Kp10 (5 µg/kg body weight (b.w.)), gonadotoropin‐releasing hormone (GnRH, 1 µg/kg b.w.), or 2 mL of saline as a control at the ages of 3 (pre‐pubertal) and 6 (post‐pubertal) months. A single i.v. injection of Kp10 significantly stimulated the release of LH and T in both groups. The area under the response curve (AUC) of LH for a 60‐min period after the i.v. injection of Kp10 was significantly greater in the pre‐pubertal goats (P < 0.05). The AUC of T for a 120 min period post‐injection did not differ between the two age groups. A single i.v. injection of GnRH also significantly stimulated the release of LH and T in both groups (P < 0.05). The secretory pattern of LH and T in response to GnRH resembled that in response to Kp10. These results show that the LH‐releasing response to Kp10 is greater in pre‐pubertal than post‐pubertal male goats. They also show that Kp10, as well as GnRH, is able to stimulate the release of T in male goats.     

Effect of supportive therapy on the pharmacokinetics of intravenous marbofloxacin in endotoxemic sheep

The purpose of this study was to determine the influences of supportive therapy (ST) on the pharmacokinetics (PK) of marbofloxacin in lipopolysaccharide (LPS)-induced endotoxemic sheep. Furthermore, minimum inhibitory concentration (MIC) of marbofloxacin against Escherichia coli, Mannheimia haemolytica, Pasteurella multocida, Klebsiella pneumoniae, Salmonella spp., and Staphylococcus aureus was determined. The study was performed using a three-period cross PK design following a 15-day washout period. In the first period, marbofloxacin (10 mg/kg) was administered by an intravenous (IV) injection. In the second and third periods, marbofloxacin was co-administered with ST (lactated ringer + 5% dextrose + 0.45% sodium chloride, IV, 20 ml/kg, dexamethasone 0.5 mg/kg, SC) and ST + LPS (E. coli O55:B5, 10 µg/kg), respectively. Plasma marbofloxacin concentration was measured using HPLC-UV. Following IV administration of marbofloxacin alone, the , AUC_0–∞, Cl_T, and V_dss were 2.87 hr, 34.73 hr × µg/ml, 0.29 L hr⁻¹ kg⁻¹, and 0.87 L/kg, respectively. While no change was found in the MBX + ST group in terms of the PK parameters of marbofloxacin, it was determined that the Cl_T of marbofloxacin decreased, AUC_0–∞ increased, and and MRT prolonged in the MBX + ST + LPS group. MIC values of marbofloxacin were 0.031 to >16 µg/ml for E. coli, 0.016 to >16 µg/ml for M. haemolytica, 0.016–1 µg/ml for P. multocida, 0.016–0.25 µg/ml for K. pneumoniae, 0.031–0.063 µg/ml for Salmonella spp., and 0.031–1 µg/ml for S. aureus. The study results show the necessity to make a dose adjustment of marbofloxacin following concomitant administration of ST in endotoxemic sheep. Also, the PK and pharmacodynamic effect of marbofloxacin needs to be determined in naturally infected septicemic sheep following concomitant administration of single and ST.     

Chemerin analog regulates energy metabolism in sheep

Accumulating data suggest a relationship between chemerin and energy metabolism. Our group previously described gene cloning, expression analysis and the regulatory mechanism of chemerin and its own receptor in mice and cattle. The objective of the present study was to investigate the physiological effect of chemerin on endocrine changes and energy metabolism in sheep using a biologically stable chemerin analog. The chemerin analog was intravenously administrated (100 or 500 µg/head) to sheep, and plasma insulin and metabolites (glucose, nonesterified fatty acids (NEFA), triglyceride, total cholesterol and high‐density lipoprotein (HDL) cholesterol) were analyzed. The chemerin analog dramatically increased the insulin levels, and glucose levels were decreased. NEFA levels were slightly decreased at 20 min but then increased gradually from 60 to 180 min after analog administration. In addition, injection of the chemerin analog immediately increased triglyceride and total cholesterol but not HDL levels. These results suggested that chemerin analog regulated insulin secretion related to glucose metabolism and the release of triglycerides in sheep in vivo. This study provides new information about endocrine and metabolic changes in response to chemerin in sheep.     

Expression of melatonin and its related synthase and membrane receptors in the oestrous corpus luteum and corpus luteum verum of sheep

Melatonin is an important factor involved in regulating reproduction; it is synthesized enzymatically by the sequential action of melatonin‐synthesizing enzymes, arylalkylamine N‐acetyltransferase (AANAT) and hydroxyindole‐O‐methyltransferase (HIOMT), and exerts its biological functions mainly through receptor‐mediated action. To evaluate the expression of melatonin, two melatonin‐synthesizing enzymes (HIOMT and AANAT), and membrane receptors (MT1 and MT2) in oestrous corpus luteum (CL) and CL verum of sheep (Ovis aries), we performed ELISA, qRT‐PCR, western blotting and immunohistochemistry. The quantitative results showed that melatonin, HIOMT and AANAT levels in the CL verum were significantly higher than those in oestrous CL (p < 0.05), whereas MT1 and MT2 exhibited no change between the oestrous CL and CL verum (p > 0.05); moreover, the localization results showed that HIOMT, AANAT, MT1 and MT2 were mainly expressed in large luteal cells (LLCs). In summary, the above results suggested that sheep CL has potential for the synthesis of melatonin; meanwhile, they also suggested that CL is one of the targets of melatonin. These results provide not only a basis for whether sheep CL can synthesize melatonin but also provide a reference for further study on the mechanism of melatonin in the CL.     

Effects of intracerebroventricular infusions of corticotropin‐releasing hormone in sheep

To provide new insights into the neural mechanisms of physiological and behavioral responses to stressors in sheep, acute changes in endocrine, autonomic and behavioral functions following 30 min infusions of ovine‐corticotropin‐releasing hormone (oCRH; 0, 0.5, 5 or 50 µg/0.5 mL of artificial cerebrospinal fluid/30 min) into the third ventricle of sheep (n = 7–8) were examined. Serial blood samples were collected through indwelling jugular catheters to determine plasma cortisol concentrations (CORT). Heart rate (HR) and rectal temperature (RT) were obtained via telemetry systems. The behaviors of the animal were monitored simultaneously. Intracerebroventricular infusions of CRH dose‐dependently induced an increase in CORT; there was a time–treatment interaction in CORT (P < 0.001). There was not a time–treatment interaction either in HR (P = 0.29) or in RT (P = 0.28). That RT showed a tendency to decrease with higher doses of CRH in sheep was in contradiction to previous reports in rats and pigs. As to changes in behavioral function, only the induction of bleating was marked. These results suggest that in physiological and behavioral responses of sheep to stressors, CRH regulates the increase in CORT and the induction of bleating. However, CRH might have little function in sympathetic nervous activation during physiological responses to stressors in sheep.     

Species Differences in the Susceptibility of Erythrocytes Exposed to Free Radicals In Vitro

The susceptibility of human, cow, pig, sheep and rabbit erythrocytes to free radicals (peroxyl radicals) generated in vitro by 2,2-azo-bis(2-amidinopropane) hydrochloride (AAPH) was evaluated by means of a haemolysis test and expressed as the time to 50% of maximal haemolysis (HT₅₀). The most sensitive to damage by free radicals appeared to be the erythrocytes of pigs and sheep, their HT₅₀ values being (mean±SEM) 85.1±1.28 min and 89.0±1.31 min, respectively. Human erythrocytes and those of cows and rabbits were about twice as resistant, their HT₅₀ values being (mean±SEM) 174.3±1.53 min, 181.2±1.22 min and 183.4±2.54 min, respectively. Pig and sheep erythrocytes used in the haemolysis test provided an indication of the antioxidant status in a shorter time (2.5 h versus 4.5 h) than those of the other species studied. The results indicate that the HT₅₀ test may be a convenient alternative to the osmotic resistance test for defining the antioxidant resistance of erythrocytes.     

Effect of intracerebroventricular injections of prolactin‐releasing peptide on prolactin release and stress‐related responses in steers

Some evidence suggests that there might be a species difference in the effect of intracerebroventricularly administered (ICV) prolactin‐releasing peptide (PrRP) between rodents and sheep. We compared the levels of cortisol (CORT) and prolactin (PRL), rectal temperature (RT) and behavioral responses to ICV bovine PrRP (bPrRP) in steers. ICV bPrRP (0.2, 2 and 20 nmol/200 µL) tended to evoke a dose‐related increase in CORT concentrations and 0.2 nmol of bPrRP induced transient increase in PRL concentrations. A significant time–treatment interaction was observed for the percent change of CORT (P < 0.05) and PRL (P < 0.05) from pre‐injection value. The time–treatment interaction for changes in RT was not significant (P = 0.50). There tended to be a difference among the four treatments in terms of maximum change in RT from the pre‐injection value between 0 and 90 min (P < 0.1). Stress‐related behavioral signs were not observed in the present experiment. These findings indicate that ICV bPrRP increased CORT and PRL levels, suggesting that central PrRP might participate in controlling the hypothalamo‐pituitary‐adrenal axis and PRL release in cattle, unlike sheep. In contrast, central PrRP is unlikely to be involved in controlling the behavior of this species because ICV bPrRP did not induce marked changes in their behavior.     

Immunomodulatory effects of Alliums and Ipomoea batata extracts on lymphocytes and macrophages functions in White Leghorn chickens: In vitro study

We previously described that supplementary garlic, onion and purple sweet potato (PSP) enhance humoral immune response in White Leghorn chickens. In the present in vitro study, we investigated the effects of garlic (GE), onion (OE) and PSP (PSPE) extracts on proliferation, interleukin (IL)‐2 and interferon (INF)‐γ gene expression of stimulated lymphocytes. The effects on microbicidal activity, reactive oxygen species (ROS) and nitric oxide (NO) productions of stimulated peritoneal macrophages were studied as well. The results showed that GE augmented Concanavalin A (ConA)‐induced splenocytes (4, 8 and 16 µg/mL) and thymocytes (2, 4 and 8 µg/mL) proliferations, and gene expression of IL‐2 (8 and 16 µg/mL) and INF‐γ (16 µg/mL). None of the examined extracts had mitogenic effect nor stimulated bursacytes response to phorbol 12‐myristate 13‐acetate (PMA). Macrophages exhibited superior microbicidal activity and ROS production with GE at 4 and 8 µg/mL and with OE at 25.6 µg/mL. None of the extracts showed stimulatory effects on NO production. The extracts showed concentration‐dependent inhibitory effects on all measured parameters at higher concentrations. Taken together, it is likely that garlic has direct stimulatory effects on immune cell functions, whereas the in vitro inhibitory effects of onion and PSP were likely attributed to high flavonoid contents.     

The effects of maternal plasma dobutamine levels on fetal oxygenation in anaesthetized sheep

Objective To measure the effects of dobutamine infusion on fetal oxygenation during isoflurane anaesthesia in pregnant ewes. Study design Prospective randomized experimental study. Animals Seven clinically normal adult pregnant Rambouillet‐Dorset cross ewes with fetuses of 117–122 days gestational age. Methods The ewes were anaesthetized with ketamine (2 mg kg^?1) IM, and isoflurane (F_E′ISO 2.0%) in oxygen. After instrumentation and stabilization, dobutamine was infused at 4 µg kg^?1minute^?1 for 60 minutes and 10 µg kg^?1minute^?1 for 60 minutes in random order, separated by a 20‐minute washout period. Catheters were placed in the maternal and fetal carotid arteries; these were used for continuous blood pressure measurement and intermittent blood sampling. Results Maternal mean systemic carotid arterial pressure was 60 mm Hg prior to dobutamine infusion. After 5 minutes of dobutamine infusion, fetal oxygen saturation increased (p < 0.05) from 0.62 (0.17–0.71, minimum–maximum) to 0.72 (0.28–0.78) at a dose of 4 µg kg^?1minute^?1 and to 0.70 (0.20–0.73) at a dose of 10 µg kg^?1minute^?1. These increases were maintained during the infusion and were not significantly different between doses. Maternal oxygen saturation remained constant at 1.0 before and during all infusions. Although maternal heart rate and blood pressure increased (p < 0.05) by 90% and 25%, respectively, with dobutamine, this stimulant effect was not evident in the corresponding fetal variables. Maternal haemoglobin concentration increased 30% (p < 0.05) with each infusion. Conclusions Dobutamine at 4 µg kg^?1minute^?1 increases fetal oxygenation that is not improved by a dose of 10 µg kg^?1minute^?1. This increase is largely due to an increase in maternal haemoglobin concentration that, in turn, increases oxygen delivery to the placenta. Clinical relevance The use of dobutamine to treat hypotension in pregnant sheep during isoflurane anaesthesia improves fetal oxygenation. This may be true in other species.     

Effects of acepromazine, butorphanol and buprenorphine on thermal and mechanical nociceptive thresholds in horses

Reasons for performing study: To investigate the antinociceptive effects of buprenorphine administered in combination with acepromazine in horses and to establish an effective dose for use in a clinical environment. Objectives: To evaluate the responses to thermal and mechanical stimulation following administration of 3 doses of buprenorphine compared to positive (butorphanol) and negative (glucose) controls. Methods: Observer blinded, randomised, crossover design using 6 Thoroughbred geldings (3–10 years, 500–560 kg). Thermal and mechanical nociceptive thresholds were measured 3 times at 15 min intervals. Horses then received acepromazine 0.05 mg/kg bwt with one of 5 treatments i.v.: 5% glucose (Glu), butorphanol 100 µg/kg bwt (But) buprenorphine 5 µg/kg bwt (Bup5), buprenorphine 7.5 µg/kg bwt (Bup7.5) and buprenorphine 10 µg/kg bwt (Bup10). Thresholds were measured 15, 30, 45, 60, 90, 120, 150, 180, 230 min, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 24 h post treatment administration. The 95% confidence intervals for threshold temperature (ΔT) for each horse were calculated and an antinociceptive effect defined as ΔT, which was higher than the upper limit of the confidence interval. Duration of thermal antinociception was analysed using a within‐subjects ANOVA and peak mechanical thresholds with a general linear model with post hoc Tukey tests. Significance was set at P<0.05. Results: Mean (± s.d.) durations of thermal antinociception following treatment administration were: Glu 0.5 (1.1), But 2.9 (2.0), Bup5 7.4 (2.3), Bup7.5 7.8 (2.7) and Bup10 9.4 (1.1) h. B5, B7.5 and B10 were significantly different from Glu and But. No serious adverse effects occurred, although determination of mechanical thresholds was confounded by locomotor stimulation. Conclusions: Administration of acepromazine and all doses of buprenorphine produced antinociception to a thermal stimulus for significantly longer than acepromazine and either butorphanol or glucose. Potential relevance: This study suggests that buprenorphine has considerable potential as an analgesic in horses and should be examined further under clinical conditions and by investigation of the pharmacokinetic/pharmacodynamic profile.     

Perineal analgesic actions of epidural clonidine in cattle

Objective To determine the analgesic, sedative, motor, cardiac and respiratory effects of epidural clonidine in cattle. Study design Prospective randomized study. Animal population Six healthy male cattle weighing between 236 and 365 kg. Methods To investigate the effect of epidural clonidine, the animals received 2 and 3 µg kg^?1 of clonidine diluted to 8 mL with 0.9% saline. Two treatments were utilized as controls. The animals from the first control treatment received 2% lidocaine (0.4 mg kg^?1) and those from the second received an equal volume of 0.9% saline. Each animal received each treatment in random order. Evaluations of analgesia, sedation, muscle relaxation, heart rate, respiratory rate and rectal temperature were obtained at 0 (basal), 2, 5, 10, 15 and 30 minutes after epidural injection, and then at 30‐minute intervals until loss of analgesia occurred. All the animals received a standard noxious stimulus consisting of needle insertion into the skin and deep muscle; a 4‐point scale was used to score the response. A second scale was used to score sedation and a third for muscle relaxation. Results Both doses of clonidine were effective in producing analgesia of the tail, perineum, and upper hindlimb. Complete analgesia was present before (mean ± SE = 9 ± 4 vs. 19 ± 9 minutes) and lasted longer (311 ± 33 vs. 192 ± 27 minutes) for the 3 µg kg^?1versus the 2 µg kg^?1 dose, respectively. A dose‐dependent sedative effect of clonidine was also observed, with a peak effect between 60 and 180 minutes. No effects on heart or respiratory rates were observed with either dose of clonidine. Conclusions Epidural administration of 2 and 3 µg kg^?1 of clonidine in cattle in this study provided bilateral perineal analgesia/anesthesia with a dose‐dependent onset and duration of action. Clinical relevance Further studies are required to determine whether the analgesia is sufficient for surgery.     

Measurement of carbonic anhydrase isozyme VI (CA‐VI) in swine sera,colostrums, saliva,bile, seminal plasma and tissues

Swine secretory carbonic anhydrase VI (CA‐VI) was purified from swine saliva and an antibody to CA‐VI was generated. A specific and sensitive enzyme‐linked immunosorbent assay (ELISA) has been developed for the measurement of swine CA‐VI. The assay can detect as little as 5 ng/mL of swine CA‐VI. Typical standard curves were determined for a range of CA‐VI solutions (7.8 to 500 ng/mL). The coefficients of variation for these solutions were less than 5%. When 500, 250 or 100 ng/mL of swine CA‐VI was added to swine sera, the recoveries were 102.0%, 109.7% and 100.2%, respectively. The concentrations of CA‐VI in the saliva (26.2 ± 30.4 µg/mL), sera (3.3 ± 4.9 ng/mL), bile (153.0 ± 114.0 ng/mL), seminal plasma (124.0 ± 39.0 ng/mL) and parotid gland (441.3 ± 90.0 µg/g wet tissue), submaxillary gland (88.1 ± 124.4 µg/g wet tissue), sublingual gland (58.6 ± 24.6 µg/g wet tissue) and gallbladder (2.4 ± 1.3 µg/1g wet tissue) were determined by ELISA. The concentration of CA‐VI in colostrum was 163.3 ± 101.4 ng/mL and did not decrease within 10 days following parturition. An immunohistochemical reaction to anti‐CA‐VI antiserum was observed in the columnar epithelial cells lining the gallbladder. These data suggest that secretory CA‐VI plays various roles in pH regulation and the maintenance of ion and fluid balance.     

Antinociceptive effect of intrathecally applied alpha2 agonists (xylazine and detomidine) in sheep and the response to atipamezole

This study evaluated the antinociceptive and physiologic effects of xylazine (X) and detomidine (D) administered intrathecally (IT) at the lumbosacral space, before and after the injection of atipamezole (A) IV. The study was approved by the National Animal Protection Authorities. Five adult healthy female sheep were anaesthetized with propofol on four occasions to inject the following treatments IT: groups 1 and 2, 0.05 mg kg^?1 X (2 mg mL^?1 saline) IT; groups 3 and 4, 0.01 mg kg^?1 D (0.5 mg mL^?1 saline) IT ( Waterman et al. 1988 ). Nociceptive threshold (TH) was tested by applying pulsed and stepwise enhanced direct current ( Ludbrook et al. 1995 ) at one hind leg pastern and noting the current at the moment of foot lift. Maximum current applied was 40 mA. Baseline TH was measured twice before anaesthesia and every 10 minutes when the sheep regained consciousness. Atipamezole was given IV immediately after reaching maximum analgesic action of X and D as defined by two equal or decreasing TH values and measurements were continued for 90 minutes. The dose of A for groups 1 and 3 was 0.005 mg kg^?1 (0.25 mg mL^?1 saline) IV, and for groups 2 and 4 was 0.0025 mg kg^?1 A (0.25 mg mL^?1 saline) IV. Heart rate (HR), mean direct arterial pressure (MAP), PaO₂ and PaCO₂ were measured. The differences between measurements recorded before and after treatment were analysed using a paired t‐test for the drug effects and a nonparametric Wilcoxon's rank sum test for the comparison between groups. A p‐value < 0.05 was considered significant. All sheep were able to stand before A IV. Threshold baseline value was 4.5 ± 1.7 (mean ± SD) mA for all animals. Xylazine caused a significantly higher TH rise (35.2 ± 1.8 mA), faster onset (21.1 ± 16.0 minutes) and longer duration of the TH enhancement (104.1 ± 8.6 minutes) than D (TH: 16.3 ± 7.8 mA, onset: 49.5 ± 28.4 minutes, duration: 59.3 ± 27.3 minutes). A significant increase in PaCO₂ was observed in the X and D treated animals, 0.39 ± 0.21 kPa (2.9 ± 1.6 mm Hg) and 0.39 ± 0.29 kPa (2.9 ± 2.2 mm Hg), respectively. Heart rate was significantly decreased by ?21 ± 17 beats minute^?1 for X animals and ?13 ± 13 beats minute^?1 for D. Mean arterial pressure (?9 ± 13 mm Hg for X and ?1 ± 11 mm Hg for D animals) and PaO₂ 0.65 ± 1.32 kPa (4.9 ± 9.9 mm Hg) for X and 1.45 ± 4.19 kPa (10.9 ± 31.4 mm Hg) for D animals) did not change significantly. The nociceptive threshold was not affected by A in any group. Threshold values of all X treated animals before A was 39.3 ± 1.4 mA and after was 37.2 ± 6.3 (group 1) and 40 ± 0 (group 2). Threshold values of all D treated animals before A was 21.0 ± 8.3 and after was 19.4 ± 7.3 (group 3) and 24.8 ± 8.0 (group 4). At the dosages administered intrathecally in this study, X and to a lower degree D induce antinociception without major physiologic changes. Atipamezole up to 0.005 mg kg^?1 IV does not affect the resulting antinociception as assessed by electrical stimulation.     

The relationships between the myocardial bridge and ramus interventricularis paraconalis characteristics in lamb and sheep

The myocardial bridge (MB) is an anomaly that the myocardial fibres cover on a segment of the subepicardial coronary arteries or their branches in domestic animals and humans. The aim of the present study was to determine the relationships between the characteristics of the MB and ramus interventricularis paraconalis at three levels in lambs and adult sheep. Thirty-three hearts (16 lambs and 17 sheep) were used to determine the MB (length, angle and thickness) and vessel (vessel diameter and thicknesses of tunica intima et media of ramus interventricularis paraconalis) characteristics. Independent-samples t test was applied to compare variables between lambs and sheep. Spearman's correlation analysis was conducted to evaluate the relationships between bridge and vessel characteristics at three bridge levels. Length, angle and thickness of myocardial bridges were not significantly different between the lambs and sheep (p > .05). The mean length, angle and thickness were 24.9 ± 16.1 mm, 113.7 ± 11.2° and 1,098 ± 555 µm in 33 hearts, respectively. In lambs, the mean vessel diameters were 1,930 ± 742 µm (1,534–2,325 µm), 1,247 ± 665 µm (893–1,601 µm) and 865 ± 172 µm (774–957 µm) at the pre-bridge, bridge and post-bridge levels, respectively. In sheep, the mean vessel diameters in the same order were 1,861 ± 1,068 µm, 1,337 ± 308 µm and 1,287 ± 549 µm. The bridge prevalence was 100% in the samples examined. In conclusion, coronary arterial diseases related to myocardial bridge should not be expected in sheep for veterinary cardiology practice. It may also be concluded that the cross-breeds of the Awassi and Chios sheep may be useful in experimental studies related to myocardial bridge surgery.     

Thermal and mechanical nociceptive threshold testing in pregnant sheep

ObjectiveAnalgesic regimes were compared in pregnant ewes after laparotomy by measuring thermal (TT) and mechanical (MT) nociceptive thresholds.Study designProspective randomised experimental study.AnimalsPregnant ewes at 121 days gestation underwent laparotomy as part of another research project.MethodsThermal and mechanical thresholds were measured before, and 2, 6, 24 and 48 hours after surgery. Thermal stimuli were delivered to the lateral aspect of the metatarsus via a skin-mounted probe, and mechanical stimuli to the contralateral site via a pneumatically driven 1.5 mm diameter pin. Each test was performed five times, alternating thermal and mechanical stimuli, with ten minutes between thermal stimuli. At the end of surgery ewes received either: 75 μg hour^?1 transdermal fentanyl patch (medial thigh) (group FP) (n = 8), or 3 μg kg^?1hour^?1 intra-peritoneal medetomidine via an osmotic pump (group IPM) (n = 8) inserted immediately prior to closure. Data were analysed using the Kruskal–Wallis RS Test (p < 0.05). Once a significant effect was identified, pairwise comparisons were performed using paired Wilcoxon RS tests. To compensate for multiple hypotheses testing, p < 0.005 was considered significant.ResultsPrior to surgery mean ± SD TT was 56.1 ± 5.0 °C (FP) and 55.6 ± 5.0 °C (IPM); MT was 5.3 ± 2.6 N (FP) and 8.0 ± 5.0 N (IPM). In FP there was no significant change in either TT or MT over time. In IPM there was no significant change in MT over time but TT increased at two hours to 59.2 ± 3.0 °C (p = 0.003). Skin temperature (ST) ranged from 33.0 to 34.7 °C and did not change over time. There were no significant differences between groups in TT, MT or ST.Conclusions and clinical relevanceAdministration of intra-peritoneal medetomidine (3 μg kg^?1hour^?1) by an osmotic pump increases the thermal nociceptive threshold in the immediate post operative period in pregnant sheep, suggesting that this agent may have a role in providing post-operative analgesia.