Iron status, as determined by hematologic values, serum iron concentration, total iron-binding capacity, and zinc protoporphyrin concentration, was determined in 2 groups of 6 nonpregnant monkeys. Monkeys of groups 1 and 2 had 10 and 5%, respectively, of their blood volume withdrawn per week for up to 10 weeks or until blood hemoglobin concentration was less than or equal to 10 g/dl. A third group of 6 monkeys served as controls. The majority (8/12) of the monkeys became anemic (hemoglobin concentration, less than or equal to 10 g/dl) after approximately 30 to 70% (mean, 49%) of their blood volume was removed. Anemia was accompanied by decrease in serum iron concentration and percentage of transferrin saturation. Microcytosis, hypochromasia, and increased zinc protoporphyrin concentration, all hematologic characteristics of iron deficiency, developed later. The calculated iron stores ranged from 1 to 133 mg, with mean value of 51 mg. Iron-depleted monkeys had mean calculated available iron store of 20.8 mg, whereas iron-replete monkeys had mean available iron store of 114.0 mg. Changes were not observed in monkeys of the control group during the study period. None of the baseline hematologic or biochemical analytes measured were good predictors of iron stores. The diet used at the research center did not provide sufficient iron to prevent iron deficiency in most of the monkeys from which a total amount of 30 to 70% of blood volume at 5 or 10%/week was withdrawn. Studies requiring that much blood may need to be modified to include iron supplementation, reduction of sample volume, or iron replacement after termination of projects.

Hematologic reference values were determined for a captive population of 11 Mexican wolves (Canis lupus baileyi). Wolf pups from 4 to 24 weeks old had progressive age-related increases in PCV, hemoglobin concentration, mean cell volume, and RBC counts similar to those seen in domestic dog pups (C familiaris). Hematologic indices in wolves older than 24 weeks were comparable to those of the adult domestic dog; however, PCV, hemoglobin concentration, and RBC counts were higher.

Three studies were conducted to determine the efficacy of milbemycin oxime in the prevention of Dirofilaria immitis infection in dogs. Dogs were given single or multiple experimental inoculations with infective third-stage D immitis larvae and were treated with milbemycin oxime at a target dosage of 0.5 mg/kg of body weight either once or at monthly intervals at various times after inoculation. The compound was effective in preventing infection when 1 dose was administered 30 or 45 days after inoculation. Significant, but incomplete, protection was achieved when single treatments were administered 60 or 90 days after inoculation. Multiple monthly treatments beginning 60 days after inoculation appeared to provide additive effects that resulted in restoration of complete efficacy.

One hundred four heartworm-free Beagles < 1 year old were studied to determine the efficacy of ivermectin chewable tablets and of 2 other ivermectin tablet formulations against heartworm larvae. At 30 days after SC inoculation of dogs with infective Dirofilaria immitis larvae, all ivermectin formulations were given orally at dosage of 6 microgram/kg of body weight. The ivermectin chewable tablets also were given orally at dosage of 2 and 6 microgram/kg at 30 and 45 days, respectively, after injection of larvae. Replicates of 6 or 8 dogs in each study were formed on the basis of gender and body weight and, within replicates, were randomly allocated to treatment groups. At 30 days after injection of larvae, the additional dogs (in replicates of 8) were assigned to the control group and to the group given ivermectin chewable tablets at dosage of 6 microgram/kg. All dogs were housed individually. Necropsy was performed approximately 5 or 6 months after larvae were administered. In both trials, all control dogs had heartworms at necropsy (University of Illinois-geometric mean, 35.0; Florida-geometric mean, 26.1). In both trials, the ivermectin chewable tablet (6 microgram/kg) and both tablet formulations (6 microgram/kg) given at 30 days after larval injection, and the chewable formulation (6 microgram/kg) given at 45 days after larval injection were 100% effective (P < 0.01) in preventing development of induced infection with D immitis. Of 8 dogs at the University of Illinois that were given ivermectin chewable tablets (2 microgram/kg) at 30 days after larval injection, 6 had heartworms (geometric mean, 2.25; efficacy, 93.6%; P < 0.01) and 5 of 7 dogs treated similarly in Florida had heartworms (geometric mean, 4.4; efficacy, 83.3%; P < 0.05). Drug-related adverse reactions were not observed in either trial.

The effects of IV administered amiodarone, a class-III antiarrhythmic agent, on myocardial contractility, early myocardial relaxation, and hemodynamic variables were evaluated in normal canine hearts and those with infarcts. In the normal canine heart, amiodarone had important, but relatively mild, depressant effects on left ventricular contractility (assessed by maximal positive first derivative of left ventricular pressure (+dP/dt(max)) and maximal elastance (Emax)) and heart rate when given IV at a dose of 10 mg/kg of body weight. An effect on contractility or active relaxation (assessed by maximal negative first derivative of left ventricular pressure(-dP/dt(max)) and the time constant of isovolumic pressure decrease) was not identified with smaller doses. Myocardial infarction itself caused a predictable and marked depressant effect on myocardial contractility, as indicated by decreases in +dP/dt(max) ejection fraction, Emax, and-dP/dt(max), and elevation in end diastolic pressure. Additional depressive effects on contractility and active relaxation resulted when 10 mg of amiodarone/kg was administered to dogs with myocardial infarction and these effects were sufficient to worsen acute myocardial infarction-induced heart failure. Significant changes attributable to heart rate alone could not be identified. On the basis of our findings, we suggest that amiodarone administered IV should be used with caution in dogs with compromised ventricular function.

An oral vitamin E absorption test used in human beings was modified for use in horses. The most appropriate techniques with which to measure gastrointestinal tract absorption of vitamin E (alpha-tocopherol) in horses weredeveloped. Vitamin E was administered orally, and serum values of alpha-tocopherol were measured by use ofhigh-performance liquid chromatography at 0, 3, 6, 9, 12, and 24 hours after vitamin E administration. Variables included comparison of 2 dosages (45 and 90 IU/kg of body weight), routes of administration, and absorption dynamics of 3 preparations of dl-alpha-tocopherol. Absorption of the 2 doses of dl-alpha-tocopherol acetate indicated a dose response; the area under the curve at 24 hours (AUC24) was 4.3 micrograms.h/ml for the 45-IU/kg dose and 32.2 micrograms.h/ml (P < 0.01) for the 90-IU/kg dose. Maximal absorption was apparent when vitamin E was naturally consumed in grain, compared with administration of identical preparations by stomach tube or paste. In the same horses, dl-alpha-tocopherol and dl-alpha-tocopherol acetate plus polyethylene glycol had statistically similar absorption curves and both had significantly greater AUC24, compared with dl-alpha-tocopherol acetate; values for the 3 compounds were 23.6, 25.8, and 12.6 micrograms.h/ml, respectively. The AUC24 varied betweenindividual horses, but time of peak value was consistently observed between 6 and 9 hours. On the basis of the data from this study, the recommended technique for performing the oral vitamin E absorption test in horses would be administration of 90 IU of the free form of dl-alpha-tocopherol/kg, mixed in 1 L of grain to horses from which food has been withheld for 12 hours, followed by allowing the horses ad libitum access to hay immediately after administration of the vitamin E. Three baseline serum alpha-tocopherol values should be obtained within 24 hours prior to the test, with the last sample being obtained just prior to administration of the test dose of vitamin E. Heparinized plasma also may be used for this testing procedure. alpha-Tocopherol concentration should be measured at 3, 6, 9, 12, and 24 hours after vitamin E administration.

A flow cytometric procedure was evaluated to measure the oxidative burst activity (hydrogen peroxide formation) of bovine neutrophils. The method involves measuring the oxidation of intracellular dichlorofluorescin to fluorescent dichlorofluorescein (DCF). Phorbol myristate acetate (PMA) was used to perturb the neutrophil plasma membrane. The sources of variation introduced into the DCF assay were also examined. The sources of variation were attributable to the isolation of neutrophils from blood, variation between duplicate assays and duplicate flow cytometric determinations of oxidative product formation, variation in neutrophil oxidative product formation among cows, and the variation (over repeated daily and weekly neutrophil isolations) in neutrophil oxidative product formation. A final objective was to determine effects of dexamethasone on oxidative product formation, and whether differences existed between blood and mammary neutrophils in oxidative product formation. There was an increasing trend in the formation of DCF with increasing time of incubation and with increasing PMA concentration. Increasing the concentration of PMA decreased lag time and increased the rate of oxidative product formation. The increase in DCF formation was statistically significant up to a PMA concentration of 10 ng/ml. This concentration was considered optimal for bovine neutrophils. Examination of the sources of variation indicated that (i) the neutrophil isolation technique was a major source of variation (17.2 to 28.4% of the total variation), and that more than one neutrophil isolation within a cow would be required to obtain an accurate estimation of DCF formation in neutrophils; (ii) duplicate assays and duplicate readings on the flow cytometer accounted for < 0.05% of the total variation and would not be necessary when performing the DCF assay; (iii) large variation (62.4 to 70.8%) existed among cows in neutrophil oxidative product formation, indicating that any treatment being compared should be done either within or preferably repeated across a large number of cows; and (iv) the variation over repeated daily (0.3%), but not weekly (19.6%) determinations of neutrophil oxidative product formation, were small enough to allow for the evaluation of major physiologic and environmental effects. Intramuscular administration of dexamethasone (50 microgram/ kg of body weight) resulted in an approximate 80% decrease in neutrophil oxidative product formation. Oxidative product formation was 75% less for neutrophils isolated from mammary secretions when compared with neutrophils from blood. These results indicated that the DCF procedure was responsive to factors known to interfere with oxidative metabolism of bovine neutrophils.

Samples of sera were obtained from 5,725 cows in a semiclosed herd. In each of the preceding 7 years, the herd was vaccinated against bovine viral diarrhea (BVD) with killed virus. Neutralizing antibody tests were done on all samples of sera, using cytopathic virus, BVD-TGAC virus, that was antigenically distinct from the vaccine virus. Most samples of sera had high titers of neutralizing antibodies against BVD-TGAC virus. In 48 samples of sera, neutralizing antibodies were not detected against BVD-TGAC virus, but were detected against the vaccine virus. Neutralizing antibodies against selected noncytopathic BVD viruses were not detected in several samples of serum that had neutralizing antibodies against the vaccine virus and BVD-TGAC virus. Noncytopathic BVD virus was isolated from sera obtained from 3 cows < 4 years old. Two cows were available for further testing, and persistent infection with BVD virus was confirmed in both cows. The BVD viruses isolated from those cows were not neutralized by several samples of sera. Immunoprecipitation of polypeptides induced by the vaccine virus was done with selected samples of serum. Two patterns of immunoprecipitated viral-induced polypeptides were identified. One pattern was consistent with exposure of cows with live virus. The other pattern was consistent with exposure of cows with only the killed virus vaccine.

The purpose of this study was to document the effect of calfhood vaccination for Mycobacterium paratuberculosis on a serologic ELISA. Fifteen calves vaccinated with a killed paratuberculosis vaccine and 5 unvaccinated control calves were tested from the first through the fifteenth month of life. Age of vaccination ranged from 5 to 40 days. Blood samples were collected prior to vaccination and periodically thereafter. Serum antibody was analyzed by use of the ELISA. All calves were Elisa-negative prior to vaccination. Thirteen of 15 vaccinated calves became ELISA-positive between 2 and 6 months after vaccination. The unvaccinated cohort remained Elisa-negative. Widespread use of vaccine may interfere with diagnosis of paratuberculosis and with control programs that are based on serologic tests that measure humoral antibody.

Pathophysiologic effects of Ostertagia ostertagi infection and their prevention by strategic anthelmintic treatments were studied in 3 groups each of 6 steer calves. Group-1 calves were noninfected controls. Group-2 calves were inoculated with 100,000 third-stage larvae on the 1st and 28th days of the experiment and grazed on pasture initially free of contamination. Group-3 calves were on a similar regimen as those in group 2, but were also treated with ivermectin 9 days after each larval inoculation. Group-2 calves had increased plasma pepsinogen and gastrin values and decreased weight gains, and total serum protein and albumin concentrations from the 2nd week of infection onward. They were anemic at 10 to 12 weeks and had lower carcass and meat quality at slaughter. Strategic anthelmintic treatments were effective in preventing these effects and calves in groups 1 and 3 had similar performances. On the basis of our findings, high pepsinogen values were related to worm burdens, whereas high gastrin concentrations were related to gastric lesions.