Explants were prepared from skeletal muscle tissue from 5 nondystrophic pups and from 5 pups with X-linked muscular dystrophy; pups were 2 to 17 weeks old. A serial reexplant method resulted in optimal cell density with minimal fibroblast growth. Cultures were examined daily by use of phase-contrast microscopy; differentiated (postfusion) cultures were examined by electron microscopy. Moderate nuclear pleomorphism and cell clustering were observed in cultures of normal and dystrophic muscle cells. Cultures were maintained to 27 days after plating. Minimal myofilament synthesis was observed in multinucleate cells from nondystrophic and dystrophic pups, but spontaneous contraction of myotubes was not observed during this period. Differences in growth, fusion, or differentiation of myogenic cells into multinucleate cells and myotubes were not found between dystrophic and normal muscle.

Twenty-six mixed-breed (14 males, 12 females) dogswere used in a double-blind study to evaluate the effect of milbemycin oxime against naturally acquired infection with Ancylostoma caninum. Dogs were ranked and paired, on the basis of number of hookworm eggs/g of feces, and treatment was randomly assigned. Each dog was given either the study drug or placebo (1 tablet/11.4 kg [0.5 mg/kg] of body weight). Eggs per gram of feces enumeration was done on days 3 and 7 after treatment, and dogs were euthanatized on day 7. On day 3, 5 of the 13 dogs in the milbemycin-treated group had hookworm eggs in the feces (results of the McMaster test). In these dogs, mean number of eggs per gram of feces had decreased markedly (from 5,289 to 452) and, by day 7, was 114. At necropsy, 16 A caninum adults were recovered from 2 of the milbemycin-treated dogs. On day 3, 12 of the 13 dogs in the placebo-treated group had hookworm eggs in the feces. Mean number of eggs per gram of feces in these dogs decreased slightly (from 5,243 to 2,646), but did not decrease further by day 7. A mean number of 54.4 A caninum adults was recovered from 12 of the 13 placebo-treated dogs at necropsy. Milbemycin oxime had 97.8% efficacy against A caninum. Results also indicated that milbemycin oxime may be effective against Trichuris vulpis, but not against Dipylidium caninum.

The relationship of muscle and bone structure to limb weakness was examined in 60 Duroc pigs from 3 lines divergently selected for thoracic limb weakness. The lines were designated high, control, or low, with the low line having inferior thoracic limb structure. At approximately 100 kg, 10 pigs of each line and gender were scored for thoracic limb structure and movement. Right and left thoracic limbs were collected at slaughter. A computerized morphometric image analysis system was used to determine cross-sectional areas of muscles, bones, and soft tissues at levels through the brachium, antebrachium, metacarpus, and digits. The statistical model that was used to analyze the data included the effects of line, sire, gender, and side (left vs right), with weight as a covariate. Total bone area was similar for all 3 lines of pigs at all cross-sectional levels, but significant differences in muscle and other soft tissue areas were observed, including significantly greater extensor area for the antebrachium (P less than 0.001) in low-line pigs than in control- and high-line pigs, smaller total area (P less than 0.05) of the metacarpus in low-line pigs than in control and high-line pigs, and less total area of the medial digit (P less than 0.01) in low-line pigs than in control- or high-line pigs. Total area of bone and soft tissue for each cross-sectional region was significantly greater (P less than 0.05) in boars than in gilts. Side differences also were observed in total cross-sectional areas of bone and soft tissue of the antebrachium, metacarpus, and digits. Pigs selected for inferior thoracic limb structure had less total soft tissue cross-sectional areas in distal limb regions than did control- or high-line pigs. Limb weakness may be related to altered distribution of soft tissue supporting structures of the thoracic limb.

Four consecutive 6-hour urine sample collections were performed on 7 healthy adult Holstein cows fed a diet of coastal Bermuda hay with ad libitum water consumption. Urine (via indwelling urinary catheter) and venous blood samples were collected at 6, 12, 18, and 24 hours. Total 24-hour urine production for the 7 cows ranged from 4,515 to 7,130 ml/d (mean +/- SD, 5,633 +/- 946 ml/d) or 0.02 to 0.04 ml/kg of body weight/d (mean +/- SD, 0.03 +/- 0.007 ml/kg/d). Renal clearance (C) of creatinine (Cr), sodium (Na), calcium (Ca), and magnesium (Mg) varied significantly (P less than 0.05) among individuals, but did not vary significantly among the four 6-hour collection periods. Clearance of chloride (Cl) and phosphorous (P) did not vary significantly either among individuals or among the four 6-hour periods. Clearance of potassium (K) varied significantly (P less than 0.05) among individuals and among the four 6-hour periods. Creatinine clearance was significantly (P less than 0.01) correlated with CCl, CCa, CP, and CMg when all data were considered. Significant (P less than 0.05) correlations were also found between CCl, and CK, CCa, CP, and CMg; between CCa and CP and CMg; and between CP and CMg. Fractional excretion (FE) of Na, K, Cl, Ca, P, and Mg did not vary significantly among the four 6-hour periods. Fractional excretion of Na, Ca, and Mg (P less than 0.01) and K and P (P less than 0.05) varied significantly within individuals among the 6-hour periods. Mean FE values, calculated by averaging values for each of the 4 collection periods for all 7 cows, ranged from 0.05 to 0.78% for FENa; 129.33 to 670.40% for FEK; 1.23 to 6.23% for FECl; 0.17 to 4.44% for FECa; 0.36 to 1.14 for FEP; and 4.96 to 11.73% for FEMg. Linear relationships between the clearance and fractional excretion of electrolytes were observed on base-10 logarithmically transformed data for Na, Ca, P, and Mg. Linear relationship was not found between CK and FEK or between CCl and FECl.

Using a heat and sonicated Mycobacteriumparatuberculosis Cordoba antigen (COA1) and the commercial protoplasmic-antigen (PPA-3) as antigens, an ELISA for detecting goat antibodies was standardized. When 2 reference populations, 1 positive (17 goats) and the other negative (63 goats) to disease, were used, this test showed 87.5% sensitivity and 93.6% specificity for COA1, and 88.2 and 95.2%, respectively for PPA-3. Absorption with M phlei was performed; no significant differences were found for COA1, but a lower sensitivity was found with PPA-3. This test was not especially affected by cross-reactivity with other mycobacterial disease because when 9 goats with M bovis infection were included in the M paratuberculosis control group, the specificity was only slightly different for absorbed (94.4%) and nonabsorbed sera (91.7%) for COA1, and (93.1 and 94.4%, respectively) for PPA-3. This test was used to study the percentage of seropositive goats for M paratuberculosis in 3 herds with different prevalences. Among 251 goats in southern Spain (Huelva), 40% were found positive for COA1, and 41% for PPA-3. Among 242 goats studied in southern Spain (Cordoba), 10.0% were positive for COA1 and 13.0% for PPA-3. In the Canary Island population of 176 goats, 3% were positive for COA1 and 0.5% for PPA-3. According to the accuracies of both positive and negative predictions, our test could be applied to populations with high prevalence to prevent additions to the herd and to cull infected animals (with 40% prevalence, the positive and negative predictive values are 90%), and to prevent adding infected animals to populations with moderate or low prevalence.

The role of L-asparaginase (L-ASP) in limiting signs of methotrexate (MTX) toxicosis was studied. Eight dogs were randomly allotted to 2 groups of 4 dogs. All dogs were given 400 IU of L-ASP/kg of body weight IM, on day 1. On day 10, group-1 dogs were given 3 mg of MTX/kg, IV, and group-2 dogs were given 6 mg of MTX/kg, IV. All dogs were given 400 IU of L-ASP/kg, IM, 24 hours later (on day 11). One group-2 dog was euthanatized on day 16 because of severe gastrointestinal signs that were unresponsive to treatment. A second dose of MTX, identical to that given on day 10, was given on day 20 to each surviving dog, followed by L-ASP on day 21. On day 67, the 7 surviving dogs were given 3 mg gf MTK/kg, IV. Adverse reactions observed were vomiting diarrhea, and weight loss. Gastrointestinal side effects of MTX were not attenuated with L-ASP and would be a serious limitation to use of MTX administered at an intermediate dose in the treatment of lymphoma in dogs.

We determined that the protein profiles of 14 isolates from animals with hemorrhagic septicemia were relatively homogeneous and could be placed in 2 distinct groups on the basis of their country of origin. Such differences correlated with the serotypic properties of the individual isolates; hemorrhagic septicemia isolates of Asian and North American origin (Carter B) had a major protein band with an apparent molecular mass of 32 kDa, whereas those of African origins (Carter E) had a major protein band with an apparent molecular mass of 37 kDa. The possession of a major 32-kDa protein band appeared to be unique to Carter B isolates, suggesting that electrophoresis may be a useful nonserologic technique for the identification of organisms of this serotype. Other major bands with apparent molecular masses of 27, 45, and 47 kDa were shared by all strains, regardless of their serotype. The lipopolysaccharides were of low molecular mass and relatively uniform from 1 isolate to the next.

Hepatic abscesses were induced experimentally in 5 steers by inoculating Fusobacterium necrophorum via ultrasonography-guided, percutaneous catheterization of the portal vein. Hepatic ultrasonography was performed to determine the onset and progression of abscessation. Blood samples were collected before and after inoculation for performing leukocyte counts and hepatic function tests. Ultrasonographic evidence of liver abscesses was observed as early as 3 days after inoculation. Abscesses appeared as hyperechoic centers (cellular debris and pus) surrounded by hypoechoic or anechoic areas (fluid). Increases in rectal temperature, leukocyte counts, fibrinogen, globulin, bilirubin, gamma-glutamyltransferase, and sorbitol dehydrogenase concentrations were detected. Hepatic dysfunction was evidenced by decrease in serum albumin concentration and low sulfobromophthalein clearance. The ultrasonographic diagnosis of abscesses correlated well with necropsy findings.

During this investigation, the use of iohexol was compared with iotrolan for canine cisternal myelography. Iohexol and iotrolan myelography was done in 6 dogs by cisternal puncture with a 6-week interval between both procedures; each dog served as its own control. Cerebrospinal fluid (CSF) was collected for baseline analysis from each dog immediately before the contrast agent was injected. Cerebrospinal fluid samples were obtained at 1, 3, 7, and 14 days after injection of each contrast medium for cytologic and chemical analysis. Total CSF leucocyte count and glucose concentration did not change significantly in comparison with baseline data in any of the samples. After the injection of iohexol, protein concentration increased significantly in the 24-hour sample, and lactate dehydrogenase activity increased significantly in the 3-day sample. Significant difference was not found between the different samples collected at 1, 3, 7, and 14 days, compared with both contrast media. None of the dogs had seizure activity during a 5-hour postmyelographic observation period. Pathologic changes were not found by gross or microscopic examination of the spinal cord. Although a degradation in time of radiographic quality of all myelograms took place, the average radiographic score decreased more rapidly with iohexol. The average score at 90 minutes with iotrolan was comparable with the score at 45 minutes with iohexol, and the average score at 150 minutes with iotrolan was better than the score at 90 minutes with iohexol. At 5 and 10 minutes after cisternal injection, no significant difference wasobservable between the myelograms, but from 45 minutes onward, myelograms with iotrolan were superior.

An indirect immunoperoxidase staining technique (IP) is described for the detection of bovine viral diarrhea virus (BVDV) in bovine semen. The performance of the IP was compared to the reference immunofluorescent staining test in its ability to detect BVDV in 23 coded field semen samples. The IP assay which can be applied with ease to a large number of samples and does not require expensive fluorescence microscope equipment, appears to be an alternative method for BVDV detection. The IP assay can be strongly recommended for certification of BVDV-free bovine semen for artificial insemination and trading purposes and for laboratories which are not equipped for performing the immunofluorescent test.