Optimizing the Conditions for In Vitro Maturation and Artificial Activation of Sika Deer (Cervus nippon hortulorum) Oocytes

With the goal of establishing experimental protocols for cloning sika deer, various conditions for in vitro maturation (IVM) and artificial activation of sika deer oocytes were examined. In vitro maturation was evaluated in seven different culture media. The highest rate of oocyte maturation was 75.4% in 10 μg/ml follicle‐stimulating hormone (FSH), 1 μg/ml LH, 0.2 mm cysteamine and 50 ng/ml epidermal growth factor (EGF) after 24 h of IVM. The efficiency after 24 h of IVM did not differ significantly (p > 0.05) from that observed after 20 h. Cysteamine (0.2 mm ) significantly increased the maturation rates after 20 h (from 59.1% to 67.2%, p < 0.05) and after 24 h (from 63.2% to 71.6%, p < 0.05) of IVM. The IVM rates of oocytes collected during the oestrous season (75.4%) and the anoestrous season (23.3%) were significantly different at 24 h. The 20 μg/ml FSH, 2 μg/ml LH, 0.4 mm cysteamine and 100 ng/ml EGF significantly increased the maturation rates (from 23.3% to 54.2%, p < 0.01) at 24 h during the anoestrous season. For the activation experiments, the most effective method was chemical activation [ionomycin + 6‐dimethylaminopurine (6‐DMAP)], which promoted the development of sika deer oocytes to the blastocyst stage (32.4%). Our results indicate that in vitro matured sika deer oocytes are good candidates for parthenogenetic activation and that chemical treatment is needed for relatively efficient activation of the oocytes. These optimized conditions for IVM and parthenogenetic activation may be useful for efforts to restore populations of the endangered sika deer using the somatic cell nuclear transfer technique.     

P‐glycoprotein in sheep liver and small intestine: gene expression and transport efflux activity

The role of the transporter P‐glycoprotein (P‐gp) in the disposition kinetics of different drugs therapeutically used in veterinary medicine has been demonstrated. Considering the anatomo‐physiological features of the ruminant species, the constitutive expression of P‐gp (ABCB1) along the sheep gastrointestinal tract was studied. Additionally, the effect of repeated dexamethasone (DEX) administrations on the ABCB1 gene expression in the liver and small intestine was also assessed. The ABCB1 mRNA expression was determined by real‐time quantitative PCR. P‐gp activity was evaluated in diffusion chambers to determine the efflux of rhodamine 123 (Rho 123) in the ileum from experimental sheep. The constitutive ABCB1 expression was 65‐fold higher in the liver than in the intestine (ileum). The highest ABCB1 mRNA expression along the small intestine was observed in the ileum (between 6‐ and 120‐fold higher). The treatment with DEX did not elicit a significant effect on the P‐gp gene expression levels in any of the investigated gastrointestinal tissues. Consistently, no significant differences were observed in the intestinal secretion of Rho 123, between untreated control (P_eff S‐M = 3.99 × 10^?6 ± 2.07 × 10^?6) and DEX‐treated animals (P_eff S‐M = 6.00 × 10^?6 ± 2.5 × 10^?6). The understanding of the efflux transporters expression and activity along the digestive tract may help to elucidate clinical implications emerging from drug interactions in livestock.     

A multicenter, matched case-control study of risk factors for equine laminitis

Risk factors for equine laminitis were examined in a prospective case-control study of the 258 cases seen at six collaborating veterinary teaching hospitals over a 32-month period. Case-control pairs were matched on institution, clinician, and season of diagnosis. The 90% of case-control pairs (78 acute, 155 chronic) that had complete data for age, gender, and breed were used in separate conditional logistic-regression models for acute and chronic laminitis. There was an increase in risk for horses with acute laminitis from 5 to 7 years of age (OR 4.7, 95% CI 1.3–16) and from 13 to 31 years of age (OR 3.9, 95% CI 1.3–12) (both compared to <5 years); risk was increased for chronic laminitis from 10 to 14 years (OR 3, 95% CI 1.4–6.8) and from 15 to 38 years (OR 2.9, 95% CI 1.4–6.1) (both compared to <6 years). Mares — but not stallions — were more likely than geldings to develop acute laminitis (OR 2.6, 95% CI 1.1–6.2) and chronic laminitis (OR 2.0, 95% CI 1.1–3.6). In the small acute-laminitis data set, the breed variable was collapsed into three categories: Thoroughbred (THB, reference), the Quarter Horse (QH), and other (non-QH-THB). The non-QH-THB group was at increased risk of acute laminitis (OR 3.8, 95% CI 1.2–11.8). For the seven breed-group categories used in the chronic-laminitis model, however, all non-THB breed groups appeared significantly at risk as compared to the THB, with odds ratios ranging from 3.3 (95% CI 1.3–8.30) for the QH to 9.1 (95% CI 2.1–39.3) for ponies.     

Effect of hydroxyethyl starch infusion on colloid oncotic pressure in hypoproteinemic horses

OBJECTIVE: To determine the effect of hydroxyethyl starch (HES) on colloid oncotic pressure (pi) during fluid resuscitation of hypoproteinemic horses and to evaluate the clinical usefulness of direct and indirect methods for determination of pi before and after infusion of a synthetic colloid. DESIGN: Prospective clinical study. ANIMALS: 11 hypoproteinemic horses. PROCEDURE: Horses received IV infusions of 8 to 10 ml of a 6% solution of HES/kg (3.6 to 4.5 ml/lb) of body weight during fluid resuscitation. Blood samples were obtained for determination of plasma measured colloid oncotic pressure (pi meas) and plasma total protein and albumin (A) concentrations. Plasma globulin concentration (G) was calculated as the difference between plasma total protein and albumin concentrations. Calculated values for colloid oncotic pressure (piA + G) were determined by use of a predictive nomogram previously developed for horses. RESULTS: There was no significant difference between the means of pi meas and piA + G at the beginning of HES infusion. After HES infusion, the mean of pi meas was increased significantly from baseline for 6 hours. Mean plasma total protein and albumin concentrations and piA + G were decreased significantly from baseline for 24 hours. Differences between mean pi meas and piA + G after HES infusion were significant for 24 hours. CONCLUSIONS AND CLINICAL RELEVANCE: There was good agreement between plasma pi meas and piA + G in blood samples obtained from hypoproteinemic horses immediately before infusion of HES. Use of a predictive nomogram did not, however, account for the oncotic effect of HES. Results of comparison of pi meas to piA + G after HES infusion suggest that a significant oncotic effect was maintained for 24 hours in the study horses.     

Morphine, pethidine and buprenorphine disposition in the cat

Pharmacokinetics of morphine, buprenorphine and pethidine were determined in 10 cats. Six cats received morphine (0.2 mg/kg) intravenously and four intramuscularly. Five received buprenorphine (0.01 mg/kg) intravenously and six intramuscularly. Six received pethidine (5 mg/kg) intramuscularly. Jugular venous blood samples were collected at time points to 24 h, and plasma morphine concentrations were measured by high performance liquid chromatograpy (HPLC), buprenorphine by radioimmunoassay (RIA) and pethidine by gas chromatography. Our data for morphine show elimination half-life (t1/2el) 76.3 min intravenous (i.v.) and 93.6 min intramuscular (i.m.); mean residence time (MRT) 105.0 and 120.5 min; clearance (Clp) 24.1 and 13.9 mL/kg/min; and volume of distribution (V(dss)) 2.6 and 1.7 L/kg, respectively. Comparable data for buprenorphine are t1/2el 416.8 and 380.2 min; MRT 417.6 and 409.8 min; Clp 16.7 and 23.7 mL/kg/min; and V(dss) 7.1 and 8.9 L/kg. For i.m. pethidine, t1/2el 216.4 min; MRT 307.5 min; Clp 20.8 mL/kg/min and V(dss) 5.2 L/kg. For i.m. dosing, the tmax for morphine, buprenorphine and pethidine were 15, 3 and 10 min, respectively. The pharmacokinetics of the three opioids in cats are broadly comparable with those of the dog, although there is a suggestion that the cat may clear morphine more slowly.     

Cytogenetic alterations in four feline soft-tissue tumours

Four mesenchymal neoplasms (two malignant fibroblastic histiocytomas, one fibrosarcoma, one fibroma) were investigated cytogenetically. Although no specific alterations were present, the frequent nonrandom involvement of chromosome E1 became clearly evident. Karyotypic alterations involving unrelated clonal changes were also present in the fibroma.