Effects of the endophyte Acremonium coenophialum in tall fescue on pregnant mares and foal viability were evaluated. Twenty-two mature pregnant mares were randomly chosen to graze either Kentucky-31 tall fescue that was free from A coenophialum (endophyte-free, EF) or tall fescue infected with A coenophialum (endophyte-present, EP) after the first 90 days of pregnancy through parturition. Concentrations of pyrrolizidine and ergopeptine alkaloids were significantly greater in EP grass, compared with EF pasture. Ten of 11 mares grazing EP pasture had obvious dystocia. Mean duration of gestation was significantly greater for the EP group, compared with the EF group. Foal survivability was severely reduced among mares grazing Ep fescue with only 1 foal surviving the natal period. Udder development and lactation were low in mares grazing EP grass. The absence of clinical problems in mares grazing EF grass implicated the endophyte as the causative agent of reproductive problems and perinatal foal mortality in pregnant mares grazing endophyte-infected fescue grass. Caution should be exercised in allowing pregnant mares to graze pastures infected with the endophyte A coenophialum.

The purpose of the study was to determine whether there were circadian variations in serum osteocalcin in normal horses and to determine whether it was important to regulate the time of blood sampling in clinical investigations. Osteocalcin or bone Gla-protein (BGP), alkaline phosphatase, total calcium, phosphate and total protein were studied over a 24 h period. Blood samples were taken every 60 min from nine adult Standardbred horses. There was a correlation between serum levels of alkaline phosphatase (r = 0.3, p < 0.01), phosphate (r = 0.42, p < 0.01) and serum osteocalcin levels. There was a very marked individual effect on serum levels of osteocalcin and alkaline phosphatase (p < 0.01). This effect was present for phosphate levels but not significant for total calcium. The individual effect was lower and time effect was higher for serum osteocalcin if the subjects were divided into two age groups, one of horses of five years or less (n = 4) and a second group older than five years (n = 5). In both groups a circadian rhythmicity was observed. Serum osteocalcin showed a biphasic pattern. Levels were constant during daytime (light period) and underwent significant variations during the night (dark period), going through a nadir at 2000 h and through a maximum peak at 0500 h. It was concluded that in normal horses the blood osteocalcin level follows a circadian variation. Also daytime (light period) seems to be the more appropriate period for blood sampling.

Analysis of hepatic enzyme activities in serum samples from 1- to 3-day-old pups revealed alkaline phosphatase (ALP) activities that were 30 times higher and gamma-glutamyltransferase (GGT) activities that were 100 times higher than activities in clinically normal adult dogs. A study was conducted to investigate high enzyme activity in pups and to determine whether there is any association between serum enzyme activity and colostrum ingestion, passive transfer of maternal serum enzyme (in colostrum or in utero), or excessive renal or hepatic tissue enzymes. Serum enzyme activity was quantified in 15 neonatal pups before and after ingestion of colostrum and in 3 colostrum-deprived neonates fed a milk substitute. Serum samples were collected on postpartum days 0, 1, 10, 15, and 30. Enzyme activity was also quantified in serum from pregnant and lactating bitches (collected on days - 2, 0, 1, 10, 30), hepatic and renal tissue from clinically normal adult dogs and 1-day-old pups, colostrum, milk (collected on days 10 and 30), and milk replacer. Significant (P < 0.01) differences in serum GGT and ALP activities between colostrum-deprived and suckling pups did not exist before initial feeding. Significant (P < 0.001) increases in serum GGT and ALP activities developed within 24 hours in suckling pups, but not in the colostrum-deprived pups. At 10 and 30 days after birth, serum GGT and ALP activities were less than values before suckling in all pups. Enzyme activities in bitches' serum remained within the normal range for adult dogs throughout whelping and lactation. Renal GGT and ALP activities were substantially greater than hepatic enzyme activities in neonates and adults. Renal tissue from adults contained 3 times greater GGT and 2 times greater ALP activities than that from neonates. Hepatic tissue from neonates contained 5 times more GGT activity than did hepatic tissues from clinically normal adults; however, hepatic ALP activity was similar in adults and neonates. Colostrum and milk bad substantially higher enzyme activities than did bitches' serum. Activities of GGT and ALP in milk were 100 times and 10 times greater, respectively, than activities in serum through day 10. By day 30, GGT and ALP activities in milk were less than before suckling. Enzyme activity was not detected in the milk substitute. These studies reveal an association between colostrum ingestion by suckling and acute, profound increases in serum GGT and ALP in 1- to 3-day-old pups. Although this phenomenon might be useful as an indicator of colostrum ingestion, it precludes the diagnostic use of either enzyme as an indicator of hepatobiliary disease in 3-day-old pups.

The efficacy of netobimin against digestive tract strongyles and the small lungworm Muellerius capillaris was tested in a dairy goat herd. The drenches were given orally at the rate of 20 mg/kg of body weight once, 10 mg/kg on 2 successive days, or 7.5 mg/kg on 3 successive days. Fecal egg counts and larval counts were done 8 days before and on the day of drenching; further counts were performed on postdrenching days 8 and 18. Two goats were necropsied, 1 on postdrenching day 5 and 1 on day 10 in the group treated on 3 successive days. The fecal egg counts were reduced by 44 to 79% depending on dosage on postdrenching day 18; the remaining worms were Trichostrongylus colubriformis. The larval counts of M capillaris were reduced by 72 to 92%, depending on dose. The reduction was significant in all the treated groups, compared with that in the control group. The use of netobimin at the dosage of 10 mg/kg given on 2 successive days or 7.5 mg/kg given on 3 successive days might be recommended to treat goats infected with Muellerius spp and digestive tract strongyles.

Ceruloplasmin (Cp) was isolated from fresh equine plasma by precipitation, cellulose chromatography, and improved ion-exchange chromatography. Purified equine Cp is a glycoprotein having a molecular weight of approximately 115,000. In electrophoresis, equine Cp migrated to the alpha 1-globulin region, its isoelectric point was about 4.15 and consisted of about 890 amino acid residues. Serum Cp concentration was measured by use of the single radial immunodiffusion method. In clinically normal horses, the mean (+/- SD) serum Cp concentration of newborn foals was 2.87 +/- 0.40 mg/ml and that of 3-month-old foals was 5.02 +/- 0.92 mg/ml, which was similar to the adult value. It reached a peak of 6.06 +/- 0.74 mg/ml in 2-year-old horses. The Cp concentration in mares was not statistically different for the perinatal period, but it decreased immediately before and after delivery. Concentration of Cp increased at 6 days after IM administration of turpentine oil, castration, or jejunojejunostomy in adult horses, and increased to peak values twice as high as baseline values at 7 to 14 days, returning to baseline values at 28 days after treatment. We concluded that equine serum Cp is an acute-phase reactive protein increased in the intermediary or later phase of acute inflammation.

Veterinary diagnostic endocrinology laboratories frequently receive hemolyzed plasma, serum, or blood samples for hormone analyses. However, except for the previously reported harm done by hemolysis to canine insulin, effects of hemolysis on quantification of other clinically important hormones are unknown. Therefore, these studies were designed to evaluate effects of hemolysis on radioimmunoassay of thyroxine, 3,5,3'-triiodothyronine, progesterone, testosterone, estradiol, cortisol, and insulin in equine, bovine, and canine plasma. In the first experiment, hormones were measured in plasma obtained from hemolyzed blood that had been stored for 18 hours. Blood samples were drawn from pregnant cows, male and diestrous female dogs, and male and pregnant female horses. Each sample was divided into 2 equal portions. One portion was ejected 4 times with a syringe through a 20-gauge (dogs, horses) or 22-gauge (cows) hypodermic needle to induce variable degrees of hemolysis. Two subsamples of the blood were taken before the first and after the first, second, and fourth ejections. One subsample of each pair was stored at 2 to 4 C and the other was stored at 20 to 22 C for 18 to 22 hours before plasma was recovered and stored at -20 C. The second portion of blood from each animal was centrifuged after collection; plasma was recovered and treated similarly as was blood. Concentrations of thyroxine in equine plasma, of 3,5,3'-triiodothyronine, estradiol, and testosterone in equine and canine plasma, and of cortisol in equine plasma were not affected by hemolysis. Storage of bovine blood at either temperature and equine blood at 20 to 22 C caused progesterone concentrations to decrease (P < 0.05); the effect was not enhanced or diminished by hemolysis. Insulin concentration in equine blood decreased (P < 0.05) at both temperatures; this effect was exacerbated by hemolysis. In the second experiment, blood samples from horses and dogs were hemolyzed and plasma was immediately recovered and stored for 18 to 22 hours at 2 to 4 C or 20 to 22 C. Storage of hemolyzed equine plasma did not affect concentrations of progesterone, insulin, or thyroxine at either temperature. Whereas progesterone concentration was not affected in hemolyzed canine plasma, hemolysis decreased (P < 0.05) insulin concentration when plasma was stored at 20 to 22 C. These results emphasize the importance of examining effects of sample collection and handling procedures on hormone stability and the danger of extrapolating results of such studies from one species to another and from one hormone to another.

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.

The effect of age on the pharmacokinetics of chloramphenicol was determined after IV administration of chloramphenicol sodium succinate (25 mg/kg of body weight) to 6 foals at 1 day and 3, 7, 14, and 42 days of age. The disposition of chloramphenicol was best described, using a two-compartment open model in all foals at all ages evaluated. Significant age-related changes were observed in values for the major kinetic terms describing the disposition of chloramphenicol in foals; the greatest changes were observed between 1 day and 3 days of age. The mean +/- SD value for elimination rate constant (beta) for chloramphenicol in 1-day-old foals (0.131 +/- 0.06 h-1) was significantly (P < 0.005) lower than the value in 3-day-old foals (0.514 +/- 0.156 h-1), and both values were significantly (P < 0.005) lower than values for beta in 7-, 14-, and 42-day-old foals. With increasing age, the increase in the mean value for beta resulted in decrease in the harmonic mean elimination half-time (t1/2 beta) for chloramphenicol, from 5.29 hours in 1-day-old foals to: 1.35 hours in 3-day-old foals; 0.61 hour in 7-day-old foals; 0.51 hour in 14-day-old foals; and 0.34 hour in 42-day-old foals. At 1, 3, and 7 days of age, values for t1/2 beta of chloramphenicol in a premature foal born after parturition was induced with oxytocin, were considerably longer than comparable t1/2 beta values for term foals born naturally. The mean body clearance (ClB) of chloramphenicol in 1-day-old foals (2.25 +/- 0.67 ml/min.kg of body weight) was significantly lower than values in: 3-day-old (6.23 +/- 2.22 ml/min.kg; P < 0.05); 7-day-old (8.86 +/- 1.90 ml/min.kg; P < 0.0005); 14-day-old (9.63 +/- 1.63 ml/min.kg; P < 0.0005); and 42-day-old (9.68 +/- 2.76 ml/min.kg; P < 0.0001) foals. In foals of all ages, ClB of chloramphenicol in the parturition-induced premature foal was lower than the mean value for term foals born naturally. The volume of distribution (V'd[area]) of chloramphenicol decreased progressively with increasing age between day 1 and day 42, so that the mean value for 42-day-old foals (362 +/- 163 ml/kg) was less than a third the mean value for 1-day-old foals (1,101 +/- 284 ml/kg). The mean value for V'd(area) in 1-day-old foals was significantly greater than values for: 7-day-old (491 +/- 158 ml/kg; P < 0.01); 14-day-old (426 +/- 65 ml/kg; P < 0.005); and 42-day-old (362 +/- 162; P < 0.0005) foals, and the mean value for V'd(area) on day 3 was significantly (P < 0.05) greater than the mean value for V'd(area) on days 7, 14, and 42. Using dosage calculations based on mean values for the pharmacokinetic terms derived for each age group, it was predicted that to maintain plasma chloramphenicol concentration > 8 microgram/ml, chloramphenicol sodium succinate (25 mg/kg) would have to be administered at dose intervals of 10, 3, 1.5, 1.5, and 1 hours in clinically normal foals 1 day and 3, 7, 14, and 42 days, of age, respectively. It was concluded that the marked changes in the disposition of chloramphenicol detectable during the first few days of life, the variation between individuals, the potentially major effect of prematurity, and the potential for compromised liver function in septicemic foals indicate that use of drugs, such as chloramphenicol, which rely heavily on hepatic metabolic processes for elimination, should be avoided whenever possible during the early neonatal period, unless plasma concentration is monitored.

The Cooper isolate of bovine herpesvirus-1, which causes abortion in cattle, was used to construct a thymidine kinase-negative (TK-) deletion mutant virus. Twelve heifers were inoculated IV at 25 to 29 weeks of pregnancy with either TK- or thymidine kinase-positive (TK+) Cooper virus. All heifers developed fevers of 1 to 2 C during the first week after inoculation. Temperatures of TK+ inoculates were slightly higher and remained above normal a few days longer than in TK- inoculates. Viremia was detected in 5 of 6 TK+ inoculates and in all 6 TK- inoculates. More virus isolations were made from nasal and vaginal swab specimens of TK+ inoculates than from swab specimens of TK- inoculates. All heifers developed virus neutralizing antibody within 14 days after inoculation and antibody titers were similar between the 2 groups. None of the TK- inoculated heifers aborted and their calves did not have neutralizing antibody at birth. Abortion occurred in 5 of 6 heifers given TK+ virus. All aborted fetuses were infected with bovine herpesvirus-1, as demonstrated by virus isolation or detection of viral antigen in fetal tissues. These results indicate that inactivation of the TK gene reduces abortifacient activity of bovine herpesvirus-1.

Immunoglobulin values were determined in fetal and kitten sera. In the fetal and precolostral kitten sera, only IgG was detected, except in 1 case in which IgM was detected. The IgG, IgA, and IgM were transferred to the kittens through colostrum ingestion with some selectivity. Concentration of the transferred IgG, IgA, and IgM decreased significantly with half-lives of 4.15 +/- 1.29 days, 2.03 +/- 0.33 days, and 2.2 +/- 1.2 days, respectively. As a result of this decrease and increase of de novo immunoglobulin synthesis, IgG, IgA, and IgM were at their lowest values when kittens were 20 to 25 days, 14 to 20 days, and 8 to 10 days old, respectively. After their nadir was reached, IgG values increased gradually, IgA slowly, and IgM rapidly, as a result of de novo immunoglobulin synthesis. When the kittens were 90 days old, their immunoglobulin values were 80% (IgG), 7% (IgA), and 100% (IgM), compared with those of adult cats. These findings suggest that kittens that receive inadequate colostrum from their mothers will be particularly susceptible to infection after they are 5 weeks old.