Fractionation of cock spermatozoa

An account is given of the methodology for fractionation of cock spermatozoa into head and tail fractions by ultrasonication, followed by sucrose density gradient centrifugation. Quantitative estimates of DNA attested to 89.4% purity of the head fraction and low contamination of tails with heads. Recovery of protein and malic dehydrogenase (MDH) activity, following sperm fractionation, averaged 94.3% and 95.7%, respectively. Contamination of the head fraction with tails, as assessed by MDH assay, was only 4.65%, and the purity of the tail fraction was 91%. Intensive succinic dehydrogenase (SDH) activity was histochemically localised in the separated tail fraction and in the tail portion of intact spermatozoa. However, SDH activity was discernible neither in the head fraction nor in the head of intact spermatozoa.     

Isolation of Mycobacterium paratuberculosis after oral inoculation in uninfected cattle.

Feces from cows naturally infected with Mycobacterium paratuberculosis was given to 6 uninfected heifers by orogastric intubation, to determine whether ingested organisms could be passively excreted and detected by bacteriologic culture of feces (ie, false-positive result). Heifers were paired, and each pair received a different dose of feces on days 1 and 2. Fecal samples were collected from the heifers 3 times daily. Mycobacterium paratuberculosis was detected in fecal samples of all heifers within 18 hours of being given the first dose of feces. The number of colony-forming units peaked on days 3 or 4, and organisms were no longer detected by day 7. The number of colony-forming units in fecal samples from the heifers was approximately proportional to the dose given. On days 15 and 16, the experiment was repeated with feces from a second infected cow. Results were similar to those in the first experiment. All heifers remained seronegative (agar-gel immunodiffusion test and ELISA) and had negative results to the intradermal johnin test throughout the experiment. Lymph node and intestinal tissues were obtained from all 6 heifers at slaughter on day 28. Mycobacterium paratuberculosis was not isolated from mesenteric lymph nodes from the ileocecal valve region, but was isolated from ileal mucosal samples from each heifer.     

Dual-Energy X-Ray Absorptiometry and Force-Plate Analysis of Gait in Dogs With Healed Femora After Leg-Lengthening Plate Fixation

Dual-energy x-ray absorptiometry was used to measure bone mineral density of four regions in healed femora of nine dogs after fracture fixation with a leg-lengthening plate. Six to 85 months (mean, 46 months) after surgery, the bone mineral density of healed femora was not significantly different from the contralateral uninjured femora ( P >.05; power = 0.8 at Δ= 15%). Radio-lucencies around the proximal screws, apparently associated with screw loosening, were seen on radiographic views of the healed femora of three dogs. In one of these dogs, one screw in the proximal metaphysis had broken. Force-plate analysis of gait was also performed on dogs at the time of bone mineral density measurement. Peak vertical force was decreased in the pelvic limb with the healed fracture compared with the contralateral unoperated limb ( P < 0.05). Clinically apparent lameness in three dogs did not appear to be associated with altered bone mineral density and may have been caused by hip osteoarthritis, a nondisplaced hairline diaphyseal fracture, and screw loosening in conjunction with extensive post-traumatic soft tissue injury.     

Evaluation of Tissue Adhesive to Contain Axonal Regeneration in Horses

Bilateral palmar and plantar digital neurectomies were completed in 10 horses (a total of 80 neurectomies) using one of three methods: (1) simple transection (guillotine method); (2) epineural capping; (3) n-butyl cyanoacrylate injected into the epineural sheath to act as a nerve sealant. Horses were regularly evaluated clinically for tenderness in and around the surgical site, as well as skin sensation at the coronary band in the heel region, during the 12-week course of the study. None of the surgical sites exhibited any signs of drainage or infection. Horses were then euthanatized, the nerve stumps were dissected from surrounding tissues, and the length and width of the tissue mass that had formed on the end of the nerve was recorded. Longitudinal and transverse sections of the nerve endings were examined histologically for numbers of proliferating axon sprouts (neuroma formation); whether the axons had penetrated the epineurium; degree of Schwann cell proliferation; degree of chronic inflammation; extent of foreign body reaction; extent of retrograde degeneration of the nerve bundles; and amount of fibro vascular proliferation. The proportion of legs exhibiting tenderness or heel sensation did not differ significantly between the three different treatments at any of the six different times they were examined. There was no difference between the three treatments in the length or width of the fibrous tissue scar on the ends of the nerves or in the number of sprouting axons from the ends of the nerves. Of 80 nerves examined, only two nerves were not confined to the epineurium. Both these nerves had been treated by simple transection. Statistically there was more chronic inflammation and foreign body reaction in the acrylic treated nerves, but no difference in Schwann cell proliferation or retrograde degeneration between the three treatments. There was slightly less fibrovascular proliferation in the transected nerves than in those subjected to epineural capping or acrylic, but the difference was not statistically significant. The use of the tissue adhesive n-butyl cyanoacrylate to prevent the continuous growth of axons after digital neurectomy seems to offer little advantage over more traditional methods of neurectomy.     

Use of 64Copper Measurements to Diagnose Canine Copper Toxicosis

Inherited canine copper toxicosis is a serious problem in Bedlington terriers and West Highland White terriers, and may also be a problem in other less-studied breeds. Affected dogs become ill at midlife with progressive and ultimately fatal liver disease. Treatments for removal of copper and prevention of copper accumulation are available, but are most effective if begun before the dog becomes ill. Until recently diagnosis has not been available until the dog is 1 year of age, and then only by an invasive liver biopsy with determination of liver copper concentration. The authors studied the use of 64copper for early diagnosis of canine copper toxicosis. Two procedures were evaluated. The first involved measuring the concentration of 64copper in blood 24 hours after oral administration of the radioisotope. At this time, 64copper was associated primarily with ceruloplasmin secreted into the blood by the liver. This procedure is useful in the diagnosis of the human counterpart, Wilson's disease. However, the authors found it to be nondiscriminatory between affected and unaffected dogs. In contrast, the second procedure, which involved measuring 64copper excreted in stool during 48 hours after an intravenous dose of radioisotope, yielded results that differentiated most affected and unaffected dogs.     

Effects of dopamine administration on cecal mechanical activity and cecal blood flow in conscious healthy horses

Lateral cecal arterial blood flow, carotid arterial pressure, heart rate, and mechanical activity of the circular and longitudinal muscle layers of the cecal body were measured in 7 conscious healthy horses during IV infusion of physiologic saline solution for 60 minutes (control), during a 60-minute IV infusion of dopamine (at dosages of 1, 2.5, and 5 micrograms/kg/min), and for 60 minutes after IV infusion of dopamine. The mean values for lateral cecal arterial blood flow during IV infusion of dopamine at a dosage of either 1 or 2.5 micrograms/kg/min were not significantly different from the mean values for lateral cecal arterial blood flow during IV infusion of saline solution. The mean values for lateral cecal arterial blood flow, however, were significantly greater during IV infusion of dopamine at a dosage of 5 micrograms/kg/min than the mean values for lateral cecal arterial blood flow during IV infusion of saline solution. Intravenous infusion of dopamine at 1 and 2.5 micrograms/kg/min did not significantly change the mean values for carotid arterial pressure. In contrast, the mean values for carotid arterial pressure were significantly less during IV infusion of dopamine at dosages of 2.5 and 5 micrograms/kg/min than during infusion of saline solution. The mean values for heart rate were not significantly altered by infusion of dopamine at a dosage of either 1 or 2.5 micrograms/kg/min, but infusion of dopamine at a dosage of 5 micrograms/kg/min significantly increased heart rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

ULTRASONOGRAPHIC LOCALIZATION OF A CAUDAL VENA CAVA THROMBUS IN A DOG WITH LEISHMANIASIS

A dog with visceral leishmaniasis developed rear limb edema, and distension of the caudal epigastric veins. Glomerular disease with nephrotic syndrome and hypercoagulable state was diagnosed. Sonographically there was massive thrombosis of the caudal vena cava.     

Treatment of Humeroulnar Subluxation with a Dynamic Proximal Ulnar Osteotomy A Review of 13 Cases

Humeroulnar subluxation was treated surgically in 13 dogs with 18 affected elbows using a proximal osteotomy of the ulna that allowed the ulna to elongate dynamically. Distal humeroulnar subluxation was secondary to premature closure of the distal ulnar physis in 16 elbows. One distal subluxation was secondary to a radioulnar synostosis, and one proximal subluxation developed after premature closure of the distal radial physis. The mean follow-up time was 22 months. Twenty-eight percent of the elbows were judged to have excellent results, 22% good results, 50% fair results, and none was judged to have a poor outcome. The presenting lameness grade and the severity of preoperative and postoperative humeroulnar subluxation had significant correlations with the prognosis. Associated orthopedic abnormalities and complications of concurrent surgical procedures affected the outcome in several dogs. Overall, the dynamic proximal ulnar osteotomy was a simple and effective technique for the treatment of uncomplicated humeroulnar subluxation.