The effects of subcutaneous administration of a commercially available estradiol 17 beta implant on hematologic values and the chemiluminescence response of neutrophils were evaluated in 14 steers. Chemiluminescence and hematologic values were measured in treated (n = 8) and nontreated (n = 6) steers on days -14, -7, and -1 prior to implantation. Estradiol 17 beta was implanted into the treated group of steers on day 0, and blood samples were obtained from all steers on days 1, 2, 3, 4, 8, 15, 22, 29, 36, 43, and 50. The concentration of estrogen in serum was significantly (P = 0.0120) higher following implantation. Chemiluminescence and hematologic indices were not significantly affected by either implant status or serum concentrations of estrogen. The results of this study suggested that the use of implants containing estradiol 17 beta for promotion of weight gain in steers will not result in alterations of hematologic values or the neutrophil respiratory burst.

Although cats are less susceptible to infection with Dirofilaria immitis than are dogs, the possibility of severe consequences from infection or adulticidal treatment renders preventive treatment a desirable alternative in endemic areas. To evaluate the efficacy of milbemycin oxime as a chemoprophylactic agent in cats, 48 cats were inoculated with infective D immitis larvae. Single oral treatment with 2.3 mg of milbemycin oxime (0.5 to 0.9 mg/kg of body weight) at 30 or 60 days after inoculation with infective larvae gave strong but incomplete protection. Treatment at 60, as well as 90, days after inoculation with infective larvae was completely effective in preventing development of infection. A control group of inoculated, but untreated, cats was monitored biweekly for hematologic changes and for changes in parasite-specific serum antigen and antibody concentrations. Pronounced increases in total leukocyte counts and eosinophil numbers were associated with the estimated time of in vivo molting from fourth- to fifth-stage larvae. Antibody reactivity correlated with infection status, but serum antigen concentrations through 161 days after inoculation were undetectable.

A murine hybridoma monoclonal antibody (MAB), IBF9, was generated by fusing myeloma cells (P3X63Ag8.653) with spleen cells from a BALB/c mouse immunized with the canine melanoma cell line CML-10c7. Initial screening of hybridoma antibodies was performed by use of an indirect immunoperomidase assay on formalin-fixed CML-10c7 cells. The isotype of MAB IBF9 was IgG1 as determined by radial gel immunodiffusion. The antibody was tested for reactivity against a panel of formalin-fixed, paraffin-embedded normal and neoplastic canine tissues, using immunoperoxidase staining. Immunostaining was observed in melanomas (24 of 38), a few carcinomas, basal cell tumors, and cutaneous lymphosarcomas. Immunostaining was not observed in fibrosarcomas, hemangiosarcomas, hemangiopericytomas, or histiocytomas. Staining of normal adult canine tissues was limited to a few epithehal tissues and a small percentage of lymphocytes. Fetal tissues were not reactive with MAB IBF9. There were statistically significant differences in frequency of reactivity among melanomas with regard to oral vs non-oral, malignant vs benign, and mitotic indices greater than or equal to 1 vs mitotic indices < 1. Differences were not significant when tumors were compared for degree of pigmentation or histologic type. On the basis of these findings, we suggest that MAB IBF9 may be of assistance in diagnosis of nonpigmented melanomas and in assessing the malignant potential of melanomas.

Five studies were performed to determine factors affecting progesterone concentration in skim milk. Results of the first study indicated that progesterone concentration was higher in skim milk of samples kept 16 hours in an ice bath (0 C) than of those left at room temperature (21 C). In the second study, this temperature effect was found to be reversible, with skim milk progesterone concentration increasing when whole milk samples were cooled prior to centrifugation. In the third study, [3H]-labeled progesterone was used to determine the relationship between fat content of foremilk (the first milk obtained from the teats), midmilk (milk obtained through milking), and strippings (milk obtained immediately after milking machines have been removed) samples and temperature (4 C and 21 C) on the percentage of progesterone in the skim milk fraction. The relationship between percentage of butterfat and percentage of progesterone in skim milk was linear when the log of these variables was used for calculations. In the fourth study, assayable progesterone in the skim milk fraction of foremilk, midmilk, and strippings was affected by temperature. In the fifth study, a multiple-regression procedure was used to determine the amount of variation in percentage of radioactive progesterone in the skim milk fraction. Independent variables (whole milk butterfat and temperature of incubation [1, 3, 13, 22, 37, and 50 C]) and the natural log of each variable, were entered into a stepwise multiple-regression analysis. The log of the temperature and percentage of butterfat of whole milk at the time of centrifugation accounted for 89.2% (r2 = 0.892) of the variation in the log of the progesterone concentration in the skim milk fractions. The equation describing this relationship was: log percentage of progesterone in the skim milk fraction = 4.046 - 0.144 X (log of temperature of whole milk sample) - 0.688 X (log percentage of butterfat in whole milk sample). The loss of progesterone from skim milk fractions of warm whole milk samples is possibly a physical phenomenon dependent on the temperature of the sample and its percentage of butterfat. A nomograph was created to allow others to use these variables in making adjustments in progesterone concentrations.

The study reported here was part of a long-term investigation of the effects of genotype on growth, reproduction, and metabolism in cattle grazing common Bermuda grass and endophyte-infected fescue pastures. In June 1990, blood samples were collected from the tail vein of yearling heifers and steers (Angus [AA], Brahman [BB], and their reciprocal crosses [AB, BA], n = 97). Serum amylase activity was assayed enzymatically; serum Ca and Mg concentrations were determined by atomic absorption spectrophotometry. The effects of endophyte-infected fescue depended on genotype (P < 0.001). In yearlings having at least 1 Angus parent (AA, AB, BA), grazing endophyteinfected fescue was associated with higher serum amylase activity than was grazing Bermuda grass. But serum amylase activities of BB yearlings consuming either forage were similar. Moreover, for either forage, substantial differences were related to genotype (P < 0.007) and gender (P < 0.05). Angus yearlings had higher serum amylase activity than did Brahman yearlings; AB and RA yearlings had intermediate values. Heifers had higher amylase activity than did steers. The relationship among serum values of amylase, Ca, and Mg depended on forage. Yearlings consuming endophyte-infected fescue and having at least 1 Angus parent had a moderate negative correlation between serum amylase activity and Ca concentration (r = -0.53; P < 0.0005); that is, in calves of genotypes with increased amylase activity while consuming endophyte-infected fescue (AA, AB, BA), the higher the amylase activity, the lower the serum Ca concentration. However, in yearlings consuming Bermuda grass, serum amylase and Ca values were not correlated. Conversely, grazing Bermuda grass was associated with moderate positive correlation between Ca and Mg concentrations (r = 0.46; P < 0.0003), but in yearlings grazing endophyte-infected fescue, Ca and Mg concentrations were independent. The cause, pathophysiologic mechanism, and clinical importance of these effects remain to be determined. In conclusion, serum amylase activity in yearling cattle was influenced by genotype, gender, and consumption of endophyte-infected fescue. We speculate that yearlings having at least 1 Angus parent may develop a persistent subclinical derangement of the exocrine portion of the pancreas when exposed to common environmental toxins associated with endophyte-infected fescue grass, and that purebred Brahman yearlings can resist this aspect of fescue toxicosis.

Several pharmaceutical compounds were evaluated for their ability to selectively inhibit activated coagulation factor-XIII-like enzyme activity (eg, XIIIa) in pooled equine plasma. Presence of coagulation factor-XIIIa -like enzyme activity in plasma was established by assay procedures involving incorporation of the fluorescent amine compound, monodansylcadaverine, into purified casein, which served as a protein substrate. Pharmaceuticals inhibitory to coagulation factor-XIIIa -like enzyme activity were recognized by plasma gel formation of high spectrophotometric transmittance (transparency), solubility of transparent fibrin gels in concentrated urea solution, in conjunction with simultaneous depletion of native fibrinogen fractions, and production of fibrin monomer. Compounds acting primarily as anticoagulants were recognized by lack of plasma gel formation, but retaining high spectrophotometric transmittance and no detectable depletion of native fibrinogen fractions. Compounds failing to inhibit either thrombin-mediated fibrinogen-fibrin transformation (ie, coagulation) or coagulation factor-XIIIa -like enzyme activity were recognized by opaque plasma gels caused by fibrin polymerization, low spectrophotometric transmittance values, and coinciding with depletion of native fibrinogen fractions. Pharmaceuticals capable of exerting selective inhibition of coagulation factor-XIIIa -like enzyme activity were further classified as competitive inhibitors of phase 1 (carbamide) or phase 2 (terminal amine) of the transglutamination process.

The mean area and minimal diameter of 3 histochemically determined myofiber types (1, 2A, and 2B; myosin ATPase in acid buffer) were calculated in middle gluteal muscle biopsy specimens from 62 stallions, 47 Andalusians and 15 Arabians, ranging in age from 6 to 12 years. Fourteen Andalusians and 7 Arabians were untrained, and the remainder were actively endurance-trained. The 6-month training schedules involved walking, slow trotting, and cantering. Fourteen Andalusians were moderately endurance-trained, whereas the other 19 Andalusians and 8 Arabians were strongly endurance-trained. Significant differences were not recorded between untrained and endurance-trained Arabians with respect to the area (type 1, 3,194 +/- 869 micrometers(2) and 3,150 +/- 370 micrometers(2); type 2A, 3,819 +/- 890 micrometers(2) and 3,380 +/- 356 micrometers(2); and type 2B, 4,872 +/- 962 micrometers(2) and 4,417 +/- 646 micrometers(2) or minimal diameter (type 1, 52.2 +/- 7.4 micrometers and 52.8 +/- 3.1 micrometers; type 2A, 58.1 +/- 6.7 micrometers and 55.0 +/- 2.8 micrometers; and type 2B, 65.3 +/- 6.4 micrometers and 63.4 +/- 4.3 micrometers) of the 3 fiber types, nor between untrained and endurance-trained Andalusians with respect to the area (untrained, 3,990 +/- 690 micrometers(2) moderately endurance-trained, 3,882 +/- 347 micrometers(2); and strongly endurance-trained, 3,758 +/- 510 micrometers(2)) and minimal diameter (untrained, 58.1 +/- 4.7 micrometers; moderately endurance-trained, 59.7 +/- 2.7 micrometers; and strongly endurance-trained, 58.7 +/- 4.5 micrometers) of 2A fibers. Moderately endurance-trained Andalusians had larger 2B fibers (area 18.6%; minimal diameter 14.3%) than untrained Andalusians, and the minimal diameter of type-1 and type-2B fibers was greater in strongly endurance-trained Andalusians than in untrained Andalusians (9.5% for type 1; 10.0% for type 2B). These results suggest minimal effects of endurance training on the myofiber size of adult Andalusians and Arabians, but a tendency toward increased type-1 and -2B fiber size was found for Andalusians.

Epidermal growth factor (EGF)-like activity was measured in mares' colostrum and milk by radioreceptor assay. Milk samples were collected from 22 mares 1 or more times during early lactation. Samples of colostrum were taken after parturition and before the foal first suckled (presuckle), within 6 hours after the foal first suckled (postsuckle), and on days 1, 2, 4, and 8 of lactation. In the 5 mares from which milk samples were obtained at each sampling time, presuckle colostral mean EGF-like activity (17.8 ng/ml) was greatest (P < 0.05). The mean values for EGF-like activity at all other sampling times were not significantly different from each other (postsuckle colostrum, 9.7 ng/ml; day 1, 9.6 ng/ml; day 2, 8.5 ng/ ml; day 4, 8.0 ng/ml; day 8, 7.8 ng/ml).

Plasma cortisol and immunoreactive (IR)-ACTH responses to 125 microg of tetracosactrin and cosyntropin--the formulation of synthetic ACTH available in Europe and the United States, respectively--were compared in 10 clinically normal cats. After administration of tetracosactrin or cosyntropin, mean plasma cortisol concentration reached a peak and plateaued between 60 and 120 minutes, then gradually decreased to values not significantly different from baseline concentration by 5 hours. Mean plasma IR-ACTH concentration reached a maximal value at 15 minutes after administration of tetracosactrin or cosyntropin and was still higher than baseline concentration at 6 hours. Difference between mean plasma cortisol and IR-ACTH concentrations for the tetracosactrin or cosyntropin trials was not significant at any of the sample collection times. Individual cats had some variation in the time of peak cortisol response after administration of either ACTH preparation. About half the cats had peak cortisol concentration at 60 to 90 minutes, whereas the remainder had the peak response at 2 to 4 hours. In general, however, peak cortisol concentration in the cats with delayed response was not much higher than the cortisol concentration at 60 to 90 minutes. Overall, these results indicate that tetracosactrin or cosyntropin induce a comparable, if not identical, pattern of adrenocortical responses when administered to healthy cats.

The scavenging of superoxide radicals by endogenous and therapeutically administered superoxide dismutases may prevent superoxide-mediated oxidative stress leading to lipid peroxidation, membrane lysis, and cell death in a wide variety of normal and pathologic states. Simple inorganic manganous salts such as MnCl2 also have superoxide dismutase-like activity and are extremely inexpensive, compared with enzymatic superoxide dismutase preparations. In this study, we explored the use of Mn salts as antioxidant drugs. We used the percentage of inhibition of nitroblue tetrazolium reduction by superoxide as a measure of the amount of superoxide dismutase-like activity. We found concentration-related increases in superoxide scavenging activity in simple buffer solutions upon addition of 1.25, 2.5, and 5.0 microM MnSO4. To determine whether Mn salts can inhibit oxidative damage in tissues, we used an in vitro model of lipid peroxidation in ischemic and reoxygenated rat liver slices. Concentrations of 10, 100, and 1000 micromoles MnCl2/L of buffer significantly decreased indicators of lipid peroxidation believed to be initiated by intracellular superoxide. We then determined the effectiveness of MnCl2 as a superoxide scavenger in conscious horses by measuring the superoxide scavenging ability of equine plasma before and during intravenous infusions of 1.0 L volumes of 0.9% saline solution containing 0, 12.5, or 25 mM MnCl2. Plasma Mn concentrations, which were determined by atomic absorption spectrophotometry, increased as a function of time and dose. Intravenously administered MnCl2 concomitantly produced dose-related increases in superoxide scavenging ability of equine plasma at 15, 30, 45, and 60 minutes after the onset of infusion, compared with preinfusion control values. Heart rate and blood pressure of the treated horses, which were monitored to measure toxicity of MnCl2, gradually increased in both treatment groups. Clinical adverse effects of MnCl2 administration included defecation, pawing, hyperexcitability, flank watching, and sweating. The results of this study indicate that simple Mn salts may scavenge superoxide radicals in vivo with minimal adverse reactions and at a trivial cost.