Bovine haptoglobin: single radial immunodiffusion assay of its polymeric forms and dramatic rise in acute-phase sera.

Using purified bovine haptoglobin (Hp) and specific antisera, a single radial immunodiffusion (SRID) assay method has been developed to measure the serum Hp level in cattle. Bovine Hp is a highly polymerized protein showing heterogeneous molecular forms in serum. After treatment with cysteine or glutathione, Hp was partially reduced to a homogeneous form, suitable for SRID assay. This method gives values comparable to those obtained by hemoglobin-binding capacity assay, and has the advantage of being simple and convenient. Although serum Hp was not detectable in healthy cattle, it was found more than 50-fold after invasive surgery, indicating that Hp is a characteristic acute-phase protein in cattle.     

Evaluation of an enzyme-linked immunosorbent assay for diagnosis of trichinellosis in swine.

Experimental and field trials were conducted to evaluate an ELISA for its ability to detect Trichinella-infected domestic swine and to compare ELISA results with muscle-digestion test results. The ELISA used was a commercial double-antibody kit, containing an excretory-secretory antigen, and was evaluated principally for epidemiologic use. Experimentally induced infection in swine (4 groups of 3 pigs each; inoculated with 0, 50, 500 or 5,000 larvae) was detected as early as postinoculation week 4, with seroconversion of all inoculated swine by postinoculation week 8. The rate of seroconversion appeared to be affected by initial larval dose, time after inoculation, and immunocompetence of the individual host. Determination of antibody kinetics generally revealed rapidly increasing antibody titer, followed by its steady decrease in most pigs. Once seropositive, however, all pigs remained seropositive for the duration of the 10-week study. Presence of muscle larvae was confirmed in all infected pigs at termination of the study. We recognize that the experimental conditions may not be truly representative of those under which natural infection develops in pigs; however, the ELISA detected an infected pig with muscle larval density of 0.87 larvae/g of tissue. Results of a field trial (n = 310) indicated no muscle digestion test-positive pigs (35 g of diaphragm muscle digested/pig), but 3 samples tested positive by ELISA for specificity of 99.0%.     

Skeletal muscle characteristics in young trained and untrained standardbred trotters.

Muscle biopsies were taken from the middle gluteal muscle of 28 Standardbred trotters, 3-4 years of age. The 13 horses in Group T were trained consistently from 18 months of age, whereas the 15 horses in Group UT were not exposed to any systematic training before 3 years of age. Group T horses had a lower percentage of Type IIB fibres (31%) than did Group UT horses (39%). Citrate synthase (CS) activity, representing oxidative capacity, was higher in Group T (72 mmol kg-1 min-1) than in Group UT (47 mmol kg-1 min-1). Biopsies were taken from 4 horses in each group when they were foals and then annually until 3-4 years of age. Results from this study indicate that regular training of Standardbreds from 18 months of age resulted in increased CS activity and a decrease in the percentage of Type IIB fibres. This study shows that training, not growth, is the main factor that induces a high oxidative capacity and a high Type IIA/IIB fibre ratio in muscle of Standardbred trotters.     

Chronic partial obstructive urolithiasis causing hydronephrosis and chronic renal failure in a steer.

A 13-month-old Angus steer was examined with a 6-week history of lethargy, malaise and dribbling urine. Laboratory exam revealed crystalluria and poor renal function. Ultrasound revealed hydronephrosis and hydroureter. Euthanasia was chosen because of a poor prognosis for economic recovery. Necropsy demonstrated numerous calculi causing partial urethral obstruction approximately 25 cm from the end of the penis. Secondary renal changes were confirmed. Urolithiasis occurs commonly in ruminants. Secondary obstruction is usually complete with severe consequences. This is the first report of chronic partial obstructive urolithiasis resulting in endstage renal disease.     

Ruminal biohydrogenation of fatty acids from high-oleate sunflower seeds.

The objective of these experiments was to examine methods of modifying the fatty acid composition of bovine tissues. In the first experiment, four steers were fitted with duodenal fistulas and were assigned to four diets in a Latin square design. The steers were fed a control diet or the same diet containing 10% high-oleate partially crushed sunflower seeds, serum-coated sunflower seeds, and heat-treated, serum-coated sunflower seeds for 5 d. Samples of digesta and feces were collected on d 5. The inclusion of sunflower seeds (plain or serum-coated) in the diet increased (P less than .05) the digesta concentration of stearate. The percentage of stearate in the digesta and feces was increased (P less than .05) from 51 to 67% and from 64 to 74%, respectively, when steers were fed the untreated sunflower seed. The fecal concentration of oleate was increased (P less than .05) by dietary sunflower seeds in steers that were fed the serum-coated, unheated sunflower seeds. In a second experiment, heifers (four per group) were fed a corn-based control diet or diets containing 10% of high-oleate sunflower oil encapsulated with calcium alginate, either plain, coated with blood meal, or with blood meal integrated into the pellet. After 50 d on treatment, samples of perianal adipose tissue were obtained by biopsy. The fatty acid composition of the adipose tissue was not modified by the inclusion of the encapsulated oleate in the diet. In summary, limited ruminal bypass of sunflower seed oleate was accomplished with sunflower seed but not with encapsulated oleate.     

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.     

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.     

Acute selenium toxicosis in a dog

A toxic dose of selenium administered by IM injection to a 3-year-old Chihuahua resulted in pulmonary edema and death. The compound had been dispensed to the owner inadvertently in combination with a vitamin E preparation. Vitamin and mineral products often are considered safe for use in megadoses by the uninformed public. The potential danger of selenium overdosage should not be underestimated.     

Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.