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.

Two commercially available synthetic adjuvant systems, trehalose dimycolate (TDM) and TDM + monophosphoryl lipid A (MPL), were compared with Freund complete adjuvant (FCA) for the ability to stimulate antibody production in New Zealand White rabbits (Oryctolagus cuniculus). In addition, each animal was evaluated for adverse reactions. The antigen, rat liver microsomal epoxide hydrolase, was administered sc emulsified with FCA, TDM, or TDM + MPL. Serum antibody titers were stimulated with all 3 adjuvant-antigen combinations. The highest titer was produced by use of FCA; TDM + MPL produced an intermediate response, and TDM produced the lowest titer. All of the rabbits immunized with FCA developed sterile subcutaneous abscesses. Rabbits immunized with TDM or TDM + MPL developed no abscesses, and only slight reactions at the injection sites. The synthetic adjuvant system TDM + MPL is recommended for use in rabbits, considering its adequate stimulation of antibody production with minimal adverse reactions.

Progressive changes in serum enzyme activity and liver histologic features were monitored in calves fed tansy ragwort (Senecio jacobaea)-contaminated pellets. The experiments were designed to simulate natural intoxicant ingestion conditions in relationship to the dose and duration of exposure to the toxic plant to correlate early laboratory diagnostic changes with the natural progression of the disease, thereby facilitating early diagnosis and intervention by veterinary clinicians. Eight calves were fed tansy ragwort and 4 additional calves served as controls. In group 1, 4 calves were continuously fed dried tansy ragwort mixed in a pelleted feed at a 5% concentration by dry weight until terminal liver disease developed. Serum liver enzyme (alkaline phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase) activities were monitored at weekly intervals in these calves and in the 2 controls. In group 2, 4 calves were fed the same contaminated feed for only 60 days, with return to normal feed for the duration of the trial. Two additional calves served as controls. Their liver enzyme activities were monitored every other week in conjunction with percutaneous liver biopsies. All 8 calves fed tansy ragwort-contaminated pellets developed terminal hepatopathy in either a chronic pattern (n = 6) or a chronic-delayed pattern (n = 2), with the onset of a moribund state or sudden death at 11 to 17 weeks and 27 to 51 weeks, respectively. The calves were euthanatized when classic terminal signs of hepatic encephalopathy first became evident. The clinicopathologic patterns of chronic and chronic-delayed toxicoses were typical of over 5,000 cases of field tansy toxicosis diagnosed at the diagnostic laboratory. Serum glutamate dehydrogenase was the first enzyme to increase in most animals, with a short-term increase to peak values followed by a rapid return to normal. This enzyme change was followed by increases in alkaline phosphatase and gamma-glutamyltransferase. Serum enzyme changes preceded development of recognizable histologic lesions. Vacuolar changes in hepatocyte nuclei, biliary hyperplasia, and fibrosis sequentially developed in liver biopsy specimens from each animal, whereas megalocytosis was not a predominant feature until necropsy. On the basis of our finding we suggest that the optimal tests for diagnosis of pyrrolizidine alkaloid intoxication should consist of liver biopsy and determination of concurrent serum liver-enzyme activities.

The effect of various confinement conditions on physical fitness in dogs was evaluated. Eighteen 9.5- to 10-month-old female purpose-bred Beagles were maintained individually for 3 months at a time in 1 of 6 confinement conditions: Condition A--an outdoor housing area with a conventional dog house and free access to a 6.1 X 9.1-m pen; condition B--outdoor kennel with a conventional dog house and free access to a 1.8 X 6.1-m run; condition C--indoor environmentally controlled 1.2 X 3.66-m run; condition D--0.9 X 1.2 X 0.84-m conventional laboratory cage in an indoor environmentally controlled room; condition E--0.9 X 1.2 X 0.84-m conventional laboratory cage in an indoor environmentally controlled room with treadmill exercise (7 km/h at a 10% grade) for 30 min/d, 5 d/wk; condition F--0.71 X 0.86 X 0.69-m conventional laboratory cage in an indoor environmentally controlled room. During the final week of each 3-month interval, muscle succinate dehydrogenase enzyme activities and submaximal exercise heart rates (during treadmill exercise) were determined to estimate physical fitness. Also, 5 days after being moved into a different housing condition, blood samples were collected for plasma cortisol determination. The type of confinement condition for dogs had little effect on muscle succinate dehydrogenase activity, but had a modest effect on submaximal exercise heart rates of dogs. At the fifth and tenth minutes of the treadmill exercise period, heart rates of dogs maintained in the smallest cages (condition F) were higher than those of dogs maintained in outside pens and runs (conditions A and B), indicating decreased fitness in the dogs maintained in the smallest cages. Differences in heart rates were not detected among dogs in other conditions. The confinement conditions used in this study had no detectable effect on plasma cortisol concentrations. We concluded that neither cage or pen size nor a regular mandatory exercise program substantially impacted on physical fitness of laboratory confined dogs, as long as the cages complied with federal standards and guidelines. Dogs maintained in substandard cages did have modest decreases in fitness.

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.

Milk samples were collected at onset of 508 episodes of clinical mastitis on a 1,700-cow dairy farm in Michigan. Daily milk production and disease events were recorded for all cows in the herd. Despite statistical association with severity of mastitis, this association was too weak for N-acetyl-beta-D-glucosaminidase (NAGase) activity to be of great value as a prognostic test for clinical mastitis. High milk NAGase activity was significantly (P < 0.0001) associated with: increased duration of treatment; increased duration of clinical signs of mastitis; decreased daily milk production; and increased risk of the cow being culled because of mastitis. The NAGase value was combined with days in milk production, baseline milk production before mastitis onset, parity, and season of onset to predict the outcome of clinical cases as measured by the first 3 aforementioned variables. Statistical models explained little of the variability among cows in duration of treatment (R2 = 0.11), duration of clinical signs of infection (R2 = 0.11), and milk production change (R2 = 0.09).

The purpose of this study was to evaluate the effect of withdrawal of lactose from the diet for 72 hours on lactase activity in the jejunal mucosa of conventionally raised calves. The descending portion of the duodenum of six Holstein calves < 24 hours old was cannulated. The calves were fed milk except on days 5, 6 and 7 when they were given the same volume of an electrolyte solution. Sequential biopsy specimens of the proximal jejunal mucosa were obtained for three weeks and the lactase activity determined. Lactase activity was highest on day 1 and a trend toward decreased lactase activity from birth until three weeks was observed. Mean lactase activity was significantly (p < 0.05) higher for days 1, and 3 compared to days 9, 13 and 17. The withdrawal of milk and replacement by an electrolyte solution during three days had no significant effect on jejunal mucosal lactase activity in neonatal calves.

Recent studies indicate that in animals with marked cardiac hypertrophy, there is depressed function of Ca2+ sequestration by myocardial sarcoplasmic reticulum (SR) because of down regulation of the Ca2+-ATPase gene. However, in several animal models we have observed enhancement of myocardial Ca2+ sequestration in response to chronic cardiac stimulation. We tested the hypothesis that in animals with mild cardiac hypertrophy, there is enhanced Ca2+ -cycling activity by the SR Ca2+ pump and Ca2+ -release channel. Because creatine kinase activity is consistently decreased in cardiomyopathy, we also determined whether enhanced Ca2+ cycling was accompanied by down regulation or inhibition of the creatine kinase system. Mild cardiac hypertrophy was induced by volume overload; 2% salt was added to the diet of 2-week-old turkey poults for 4 weeks. Compared with age-matched controls, volume overload resulted in 14.3% increase in heart weight and 21.5% increase in heart-to-body weight ratios. The hypertrophied heart had approximately 20% increased activities of the SR Ca2+ pump and the SR Ca2+ channel. Net Ca2+ transport was increased by 16.5%. Compared with controls and in contrast to several other myocardial enzymes, creatine kinase activity was diminished in the hypertrophied hearts by 23% and creatine content was decreased by 8%. Differences between groups were not detected for lactate dehydrogenase, aspartate transaminase, and alanine transaminase. We concluded that an early adaptation of the myocardium undergoing hypertrophy in compensatory response to functional overload is an enhancement of Ca2+ Cycling activity by the Ca2+ pump and Ca2+ channel of the SR. In contrast to late-stage hypertrophy, there is no evidence for down regulation of the Ca2+-ATPase gene. However, creatine kinase activity and creatine content are diminished by mild cardiac hypertrophy.

Tear samples were collected from 1 eye of each of 40 cows, 27 sheep, 5 goats, and 5 human beings. Additionally, 10 bovine tear samples were pooled and concentrated. Spectrophotometric assays, using Micrococcus lysodeikticus, were performed on each sample to detect lysozyme activity expressed in hen egg lysozyme (HEL) equivalents. Lysozyme activity was not detected in tears of cows, but 158.8 +/- 159.3 mg of HEL/ml was detected in tears of sheep, 220.7 +/- 37.5 mg of HEL/ml in tears of goats, and 216.3 +/- 86.2 mg of HEL/ml in tears of human beings. In pooled bovine tear samples, lysozyme activity was not detected on plate assay and lysozyme protein was not detected on polyacrylamide gel electrophoresis, column chromatography, or immunoelectrophoresis with rabbit anti-bovine tear antibodies. On the basis of these observations, we concluded that the basic ocular protective mechanism in bovine tears is not lysozyme. Other antibacterial proteins such as lactoferrin, transferrin, complement, or beta-lysin may, therefore, be of primary importance in protecting the bovine eye.

The vaccine efficacy of a genetically engineered deletion mutant strain of pseudorabies virus, strain 783, was compared with that of the conventionally attenuated Bartha strain. Strain 783 has deletions in the genes coding for glycoprotein I and thymidine kinase. In experiment 1, which had a 3-month interval between vaccination and challenge exposure, strain 783 protected pigs significantly (P < 0.05) better against virulent virus challenge exposure than did the Bartha strain. The growth of pigs vaccinated with strain 783 was not arrested, whereas that of pigs vaccinated with the Bartha strain was arrested for 7 days. Of 8 pigs given strain 783, 4 were fully protected against challenge exposure; none of the pigs given strain Bartha was fully protected. In experiment 2, which had a 3-week interval between vaccination and challenge exposure, the growth of pigs vaccinated with strain 783 was arrested for 3.5 days, whereas that of pigs vaccinated with the Bartha strain was arrested for 6 days. In experiment 3, pigs with moderate titer of maternal antibodies were vaccinated twice IM or once intranasally with either strain 783 or Bartha and were challenge-exposed 3 months after vaccination. Pigs given strain 783 twice IM were significantly (P < 0.05) better protected than were the other pigs. They had growth arrest of only 6 days, compared with 9 days for pigs of other groups, and shed less virus after challenge exposure. Results of this study indicate that the vaccine based on the deletion mutant strain 783 is more efficacious than is the Bartha strain of pseudorabies virus.