Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (UTEC)-conditioned medium, and direct culturing of spermatozoa with UTEC (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with UTEC-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P < 0.05) capacitation-like changes than did the control, a modified Tyrode's medium. More (P < 0.05) spermatozoa were viable after 24 hours of UTEC coculturing than in the control incubation.

Ultrasound-guided brain biopsy of the cingulate gyrus and the head of the caudate nucleus was performed in clinically normal dogs. Dogs survived the surgery, and neurologic deficits were not detected in the 14-day postoperative period. Magnetic resonance imaging detected changes in the brain associated with biopsy in 9 dogs (90%) immediately after surgery and in 6 dogs (60%) 14 days after surgery. Fourteen days after surgery, sonography of the brain, performed through the skin overlying the burr hole, detected changes associated with biopsy in 9 dogs (90%). Histopathologic changes evident in the brain 14 days after surgery consisted of focal malacia and hemorrhage with associated subacute encephalomeningitis. Postmortem examination indicated that the biopsy specimen was accurately obtained from the desired site in 9 dogs (90%). Tissue specimens suitable for histologic examination were obtained from 10 dogs (100%). Accuracy and low morbidity of ultrasound-guided biopsy indicate that this may be a useful technique for diagnosis of focal brain disease in dogs.

Plasma catecholamine concentrations in response to onychectomy were examined in 27 cats receiving different anesthetic regimens. Each cat was anesthetized with a dissociative-tranquilizer combination, and onychectomy was performed on 1 forefoot. One week later, each cat was anesthetized with the same dissociative-tranquilizer combination plus either butorphanol or oxymorphone, and onychectomy was performed on the other forefoot. Four treatment groups were studied: tiletamine-zolazepam and tiletamine-zolazepam-butorphanol combinations were administered to group-1 cats, ketamine-acepromazine and ketamine-acepromazine-butorphanol combinations were administered to group-2 cats, tiletamine-zolazepam and tiletamine-zolazepam-oxymorphone combinations were administered to group-3 cats, and ketamine-acepromazine and ketamine-acepromazine-oxymorphone combinations were administered to group-4 cats. All drug combinations were administered IM. Central venous blood samples were drawn for catecholamine analysis after injection of drug(s), after onychectomy, and 1, 2, and 4 hours after injection. Tiletamine-zolazepam alone or tiletamine-zolazepam-butorphanol prevented epinephrine release for 2 hours after injection of drug(s). Norepinephrine concentration increased significantly (P < 0.05) from baseline after onychectomy for tiletimine-zolazepam-butorphanol and at 4 hours for tiletamine-zolazepam and tiletamine-zolazepambutorphanol. After onychectomy, there was no difference in epinephrine values between tfletamine-zolazepam and tiletamine-zolazepam-oxymorphone. Ketamine-acepromazine prevented increases in norepinephrine and epinephrine concentrations for up to 2 hours after surgery. Addition of butorphanol to ketamine-acepromazine decreased norepinephrine values immediately after onychectomy. Addition of oxymorphone to ketamine-acepromazine resulted in lower epinephrine values 4 hours after surgery.

Concentration of sulfamethazine was measured in plasma and tissues (fat, liver, kidney, spleen, lungs, and skeletal muscle) of pigs given the drug IV and PC. The plasma concentration vs time curve was best described by a 2-compartment model, with a distribution half-life of 0.46 hour and an elimination half-life of 16.9 hours. Bioavailability after oral administration was 85.8 +/- 5.3%. The tissue and plasma sulfamethazine concentration vs time data ,ere used to develop a multicompartment pharmacokinetic model of sulfamethazine disposition in pigs. Plasma and tissue concentrations of sulfamethazine in pigs were measured at various intervals after multiple oral doses of sulfamethazine, and were compared to concentrations predicted by the model. Model predictions for tissue concentrations of sulfamethazine after addition of the drug to feed (110 micrograms/g of feed for 98 days; 550 micrograms/g for 30 days) were compared to results from other studies. The model accurately predicted the number of days for sulfamethazine concentration to fall below 0.1 Kg of tissue/g (0.1 ppm. the tolerated concentration) in various tissues.

The effect of hypercapnia on the arrhythmogenic dose of epinephrine (ADE) was investigated in 14 horses. Anesthesia was induced with guaifenesin and thiamylal sodium and was maintained at an end-tidal halothane concentration between 0.86 and 0.92%. Base-apex ECG, cardiac output, and facial artery blood pressure were measured and recorded. The ADE was determined at normocapnia (arterial partial pressure of carbon dioxide [Pa(CO2)] = 35 to 45 mm of Hg), at hypercapnia (Pa(CO2) = 70 to 80 mm of Hg), and after return to normocapnia. Epinephrine was infused at arithmetically spaced increasing rates (initial rate = 0.25 micrograms/kg of body weight/min) for a maximum of 10 minutes. The ADE was defined as the lowest epinephrine infusion rate, to the nearest 0.25 micrograms/kg/min, at which 4 premature ventricular complexes occurred in a 15-second period. The ADE (mean +/- SD) during hypercapnia (1.04 +/- 0.23 micrograms/kg/min) was significantly (P < 0.05) less than the ADE at normocapnia (1.35 +/- 0.38 micrograms/kg/min), whereas the ADE after return to normocapnia (1.17 +/- 0.22 micrograms/kg/min) was not significantly different from those during normocapnia or hypercapnia. Baseline systolic and diastolic arterial pressures and cardiac output decreased after return to normocapnia. Significant differences were not found in arterial partial pressure of O2 (Pa(O2)) or in base excess during the experiment. Two horses developed ventricular fibrillation and died during normocapnic determinations of ADE. Hypercapnia was associated with an increased risk of developing ventricular arrhythmias in horses anesthetized with guaifenesin, thiamylal sodium, and halothane.

Spontaneous variations in ECG and continuous Holter monitor recordings of a colony of 31 male and 31 female cynomoigus monkeys were characterized. Electrocardiograms recorded for approximately 1 minute on 2 occasions in nonsedated monkeys were analyzed, and intervals (PR, QRS, and QT), amplitudes (P, Q, F, and T), and heart rate were determined from lead II of these tracings. In addition, Holter monitor recorders were placed on monkeys by use of carrying jackets for 16 to 24 hours of continuous recording twice during the study, and tapes were analyzed. Mean heart rate and intervals and amplitudes were similar for males and females on the first and the second recordings, Mean heart rate for males and females was 232 and 226 beats/min (bpm), respectively. The PR, QRS, and QT interval measurements, 77, 29, and 165 milliseconds, respectively, were recorded for males and 81, 30, and 162 milliseconds, respectively, were recorded for females. The P, Q, R, and T wave amplitudes were 0.16, 0.11, 0.64, and 0.28, mV respectively, for males and were 0.17, 0.10, 0.79 and 0.24 mV, respectively, for females. In addition, ventricular ectopic beats were observed in ECG from 5 females, but not in ECG from the males. Single ventricular ectopic beats were observed in 3 females for either the first or second tracing. One monkey had ectopic beats in both tracings, but in both instances, the number of ectopic beats was low (3 singles in the first and 1 in the second tracing). One monkey had runs of pairs and bigeminal beats in only the first tracing. One monkey had sporadic beats indicative of right bundle branch block morphology in both tracings. In Holter recordings, ventricular ectopic beats were identified in 47 monkeys. Ventricular ectopic beats were observed in only 1 of the 2 Holter monitor tapes for 53% of these monkeys. Most ventricular ectopic beats occurred as single beats, but pairs, ventricular tachycardia, and bigeminy also were observed. Ectopic beats were of a single morphology in 60% of the monkeys, but as many as 4 different morphologies were observed in a single tracing. Sinus arrhythmia or arrest was observed in 66% of the monkeys. Ventricular ectopic beats and sinus arrhythmia can occur without apparent cause in clinically normal monkeys. Higher prevalences of these abnormalities are identified by Holter monitoring relative to routine ECG procedures. These variables should be cautiously evaluated, because the lack of proper characterization of monkeys on test may mislead investigators as to the real importance of these findings.

We evaluated the biochemical and hemodynamic response to hypertonic saline solution plus dextran in isoflurane-anesthetized pigs infused IV with Escherichia coli endotoxin (5 micrograms/kg of body weight for 0 to 1 hour + 2 micrograms/kg for 1 to 4 hours). After 120 minutes of endotoxemia, pigs were treated with a bolus (4 ml/kg over 3 minutes) of either normal saline solution (NSS; 0.9% NaCl), or hypertonic saline solution plus dextran (HSSD; 7.5% NaCl + 6% dextran-70). Administration of HSSD significantly (P < 0.05) increased serum osmolality and concentrations of sodium and chloride for approximately 2 hours during endotoxemia. Plasma total protein concentration decreased significantly (P < 0.05) for 2 hours after treatment with HSSD, indicating hemodilution and increased plasma volume. Although HSSD transiently increased cardiac index (CI) for approximately 15 minutes, this effect was not sustained; however, the endotoxin-induced decrease in CI was ameliorated from 120 to 180 minutes. In pigs of the endotoxin + NSS group from 180 to 240 minutes, CI decreased significantly (P < 0.05), compared with baseline and control values. The endotoxin-induced increases in mean pulmonary arterial pressure and pulmonary vascular resistance were not attenuated by HSSD. At 135 minutes, total peripheral vascular resistance was transiently lower (for approx 15 minutes) in pigs treated with HSSD, compared with control pigs. The endotoxin-induced increase in plasma lactate concentration was not attenuated by HSSD, indicating continued peripheral O2 debt. We conclude that, despite sustained increases in serum osmolality and concentrations of sodium and chloride, HSSD has only transiently beneficial cardiopulmonary effects during endotoxemia in pigs.

A sensitive and specific DNA probe for detection and identification of bovine herpesvirus 4 (BHV-4) was developed. Cloned fragments from a library of HindIII fragments of the BHV-4 (DN-599) genome were labeled with 32P or digoxigenin and were tested for sentitivity and specificity in detecting viral DNA by dot-blot hybridization. Two probes were identified that detected 10 pg of purified viral DNA, and detected viral DNA in 0.001 microgram of total DNA extracted from BHV-4-infected cells. Both probes labeled with 32P and 1 labeled with digoxigenin detected viral DNA in samples prepared from cells infected with 2 prototype strains (DN-599 and Movar 33/63) and 4 field isolates of BHV-4. The DNA probes did not hybridize to total DNA prepared from uninfected bovine cells or from cells infected with BHV-1, BHV-2, alcelaphine herpesvirus 1, pseudorabies virus, or equine herpesvirus 1. One probe, labeled with digoxigenin, was tested further by dot-blot hybridization with infected cell lysates that were simply treated with sodium dodecyl sulfate and proteinase K prior to application to the membrane, avoiding extensive DNA purification procedures. This simplified procedure also resulted in specific detection of field isolates of BHV-4 and prototype strains of BHV-4.

After removal of 1 metatarsal pad and formation of a granulation tissue bed, free segmental 6- X 8-mm grafts from digital pads were sutured into recessed same-size recipient sites in the granulation tissue. In 5 dogs, the grafted area had been denervated by excision of a segment of the tibial nerve at the level of the tarsus. The grafted area was not denervated in the remaining 5 dogs. In both groups of dogs, the grafts placed around the periphery of the wound healed, blocked ingrowth of delicate epithelium from the surrounding skin, and provided a tough keratinized epithelium that covered the wound's center. As healing progressed, the grafts coalesced as the wounds contracted. Weight bearing resulted in graft expansion to provide functional weight-bearing tissue. Dogs of the denervated group had clinical and histologic evidence of collateral sensory reinnervation of the denervated area. However, with the exception of 1 dog, results of sensory nerve action potential tests indicated that reinnervation may not have been by way of regeneration across the excisional gap in the nerve. Evaluation of reinnervation of the tibial autonomous zone in 2 additional dogs revealed clinical evidence that collateral reinnervation began between 19 and 28 days after nerve excision and progressed proximad to distad. Results of sensory nerve action potential tests indicated that reinnervation may not have been via regeneration across the excision site. Results of fluorescent tracer studies did not have positive findings regarding the route of collateral reinnervation. Segmental paw pad grafts can be used effectively to provide weight-bearing tissue on a dog's limb. With local nerve damage on the distal portion of the limb, collateral innervation can grow into the area to reinnervate tissues, including pad grafts.

Existence of ultradian variation in serum progesterone concentration and the relation between progesterone and luteinizing hormone (LH) secretory patterns were investigated in nonpregnant and pregnant mares. Blood samples were taken every 15 minutes for a 24-hour period on day 8 of the estrous cycle and day 18 of pregnancy, respectively. Progesterone and LH concentrations were determined by radioimmunoassay. Progesterone was secreted in pulsatile manner in nonpregnant and pregnant mares. Luteinizing hormone also was secreted in a pulsatile manner in both groups of mares. There was little temporal relation between LH and progesterone pulses in either pregnant or nonpregnant mares.