Hydrated sodium calcium aluminosilicate (HSCAS), an anticaking agent for mixed feed, was added to the diets of growing wethers (mean body weight, 34.0 kg) and was evaluated for its ability to diminish the clinical signs of aflatoxicosis. The experimental design consisted of 4 treatment groups of 5 wethers each, consuming concentrations of 0 g of HSCAS and 0 g of aflatoxin (AF)/kg of feed (control; group 1); 20 g of HSCAS/kg (2.0%; group 2), 2.6 mg of AF/kg (group 3); or 20 g of HSCAS (2.0%) plus 2.6 mg of AF/kg (group 4). Wethers were maintained in indoor pens, with feed and water available ad libitum for 42 days. Lambs were observed twice daily and weighed weekly, and blood samples were obtained every 2 weeks for hematologic and serum biochemical analyses and for measurement of mitogen-induced lymphocyte-stimulation index. At the termination of the study, wethers were euthanatized and necropsied. Body weight gain was diminished significantly (P less than 0.05) by consumption of 2.6 mg of AF/kg of feed, whereas body weight of lambs consuming HSCAS plus AF did not differ from that of control wethers. The AF-alone treatment increased serum aspartate transaminase and gamma-glutamyltransferase activities, prothrombin time, and cholesterol, uric acid, and triglyceride values and decreased albumin, glucose, and urea nitrogen values, and urea-to-creatine ratio. A 27% decrease in lymphocyte stimulation index, increased spleen weight (as a percentage of body weight), and decreased liver weight were induced by AF-alone treatment. Results indicate that HSCAS may be a high-affinity sorbent for AF, that 2.6 mg of AF/kg of feed induces signs of aflatoxicosis in growing wethers, that lambs may not be as resistant to the effects of AF as previously thought, that 2.0% HSCAS can substantially reduce the toxic effects of 2.6 mg of AF/kg, and that sorbent compounds may offer a novel approach to the preventive management of aflatoxicosis in livestock.

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.

Pharmacokinetic determinants of spiramycin and its distribution into the respiratory tract were studied in 2 groups of calves, 4 to 10 weeks old. Group-A calves (n = 4) were used to determine pharmacokinetic variables of spiramycin after IV (15 and 30 mg/kg of body weight) and oral administrations of the drug (30 mg/kg) and to measure distribution of spiramycin into nasal and bronchial secretions. Group-B calves (n = 4) were used to determine distribution of spiramycin into lung tissue and bronchial mucosa. Spiramycin disposition was best described by use of an open 3-compartment model. Mean (+/- SD) elimination half-life was 28.7 +/- 12.3 hours, and steady-state volume of distribution was 23.5 +/- 6.0 L/kg. Bioavailability after oral administration was 4 +/- 3%. High and persistent concentrations of spiramycin were achieved in the respiratory tract tissues and fluids. Tissue-to-plasma concentration ratio was 58 for lung tissue and 18 for bronchial mucosa at 3 hours after spiramycin administration and 137 and 49, respectively at 24 hours. Secretion-toplasma concentration ratio was 4 for nasal secretions and 7 for bronchial secretions, and remained almost constant with time. Thus, spiramycin penetrates well into the respiratory tract, although the value in bronchial secretions is lower than that in lung tissues and bronchial mucosa. Calculations indicate that a loading dose of 45 mg/kg, administered IV, followed by a maintenance dose of 20 mg/kg, IV, once daily is required to maintain active concentrations of spiramycin against bovine pathogens in bronchial secretions.

A comparison of immune variables following lung sensitization with live Pasteurella haemolytica serotype 1 (Ph1)-impregnated agar beads was done in 2 separate trials. The Ph1 immune variables studied were blood bactericidal activity, serum bacteriolysis, total classical complement, and indirect hemagglutination antibody. Each trial had 16 male weanling goats: 6 controls and 10 principals. In trial 1, each goat was surgically catheterized through the trachea, then the material was deposited in a bronchus. The controls received only agar beads and the principals received agar beads impregnated with live Ph1. These goats were studied for 32 days, euthanatized, and necropsied. In trial 2, the controls were each transthoracically injected with agar beads into the left lung and the principals were similarly injected with agar beads impregnated with live Ph1. These goats were studied for 35 days, then challenge exposed transthoracically by injection of Ph1 in saline solution (1.2 X 10(7) CFU/ml) into the right lung. Four days later, they were euthanatized and necropsied. The volume of lung consolidated tissue was an excellent measure of Ph1 immunity. Principal goats generated solid protective immunity to subsequent challenge exposure because minimal or no lung consolidation was observed, whereas large volumes of lung consolidation were seen in the controls. The principal goats in trial 1 gave a weak serum indirect hemagglutination Ph1 antibody response, which was attributed to the bronchial method of depositing the Ph1. The corresponding response of the control group remained negative. The Ph1 agar beads (1 X 10(6) CFU in 0.5 ml) protected the bacteria from immediate phagocytosis and lysis as indicated by the induced pneumonic deaths of 2 principals 5 days later. Also, live Ph1 were isolated on day 32 during necropsy of respiratory tracts of 3 principals. At necropsy, no Ph1 isolates were found in the controls. Bacteriolytic activity was not induced against Ph1 in either control or principal groups in this trial. the study, but antibody titers of the principals increased to a geometric mean of 1:250 seven days after lung injection (1 X 10(5) CFU in 0.5 ml). Serum bacteriolytic titers on day 0 indicated that both principals and controls could be subgrouped to high or low subgroups on the basis of their bacteriolytic activity. The bacteriolytic activities of the controls remained unchanged during the experiment, and neither control subgroup was protected from Ph1 challenge exposure. Bacteriolytic activities of the high and low principal subgroups responded differently to Ph1 agar bead lung injection, but both principal subgroups were protected from lung challenge exposure. The low principal subgroup generated high titers of indirect hemagglutination Ph1 antibody, whereas, the high principal subgroup generated lower antibody titers. Total complement, serum bacteriolytic, and blood bactericidal profiles were similar in the principal group with high bacteriolytic activity. The immune factors that protected 2 principal subgroups did not appear to be associated with Ph1 serum bacteriolysis.

Suspensory ligaments (SL) from 32 Thoroughbreds and 32 Standardbreds were collected to evaluate the variation in muscle content with respect to age, breed, sex, limb, and use. Six transverse sections, each 3 to 5 mm thick, were obtained from each SL. Four sections were taken from the body of the SL and 1 from the midportion of each branch. Sections were stained with van Gieson picric acid-fuchsin solution, then photographed, and black-and-white slides were made from the processed negatives. The transverse-sectional area of the SL and the contained muscle were determined by use of a computer with a color monitor and a digitizing device with its associated software. The percentage of muscle was then calculated for each section, for the entire ligament, and for each horse. Results were analyzed by multiple-regression analysis and Duncan multiple-range test, using the General Linear Model of SAS. Standardbreds had 40% more muscle in their SL than did Thoroughbreds. There was no significant difference in the percentage of SL muscle among sex, age, use, individual limb, or forelimb vs hind limb. For Standardbred horses, females had significantly greater muscle area content than intact males. Also, hind limb muscle area content was significantly greater than forelimb muscle content. Thoroughbred horses between 2 and 10 years of age not in training had significantly more muscle content than horses of the same age not in training. The reasons for these differences remain unclear.

Use of microangiography is now essential for the study of microcirculation in various organs. Renal microangiographic studies have been reported in rats, rabbits, dogs, human beings, and mice. However, we could not find any report on use of the technique in cattle, despite high incidence of renal disease in that species. The perfusion technique used in mice was improved over that of our previous report, and was applied to normal and diseased bovine kidneys. For the microangiographic technique, composition of the contrast medium, pressure of the injection, duration of perfusion, and washing of kidneys with heparinized saline solution before perfusion are important. In cattle, 1- to 2-mm-thick sections of the kidneys were generally necessary to observe renal vasculature: arcuate and interlobular arteries, afferent arterioles, and glomerular capillaries. In normal bovine kidneys, the angiographic and microangiographic findings were easily recognized as normal, compared with those of normal mice. In affected bovine kidneys, which histologically represented glomerulonephritis and pyelonephritis, angiography and microangiography revealed corresponding findings.

Quantitative electroencephalography was assessed in dogs under controlled, 2% end-tidal isoflurane anesthetic conditions, and each variable at each electrode site was tested for normal distribution. With the quantitative electroencephalographic system used, 16 values for each of 21 electrode sites were evaluated. Absolute power ratios also were evaluated. The methods for quantitative electroencephalographic recording and analysis appear to be readily adaptable to the dog. Most of the data do not conform to a normal distribution. Therefore, distribution-free nonparametric statistics should be used when looking for differences under experimental or clinical conditions. Quantitative electroencephalography appears to be a sensitive noninvasive method that could be used to evaluate brain function under anesthetic, clinical, and experimental settings.

Oral keratinocytes from dogs were cultured on either collagen gels or artificial matrices at the air-liquid interface, and the expression of keratinocyte antigens and basement membrane components was determined, using various monoclonal and polyclonal antibodies. Keratinocytes grown on collagen gels expressed pemphigus vulgaris, pemphigus foliaceous, and bullous pemphigoid antigens. Diffuse, suprabasal, and superficial keratinocyte membrane differentiation antigens identified by various monoclonal antibodies also were expressed in a pattern identical to that observed in the native tissue. Laminin and type-IV collagen were deposited at the keratinocyte-collagen interface in a patchy distribution. When synthetic matrices were used, the oral keratinocytes differentiated, but to a lesser extent than cells grown on collagen gels. Antigen expression for cells grown on synthetic matrices was similar to that for cells on collagen, except for failure of the keratinocytes on synthetic membranes to express superficial cell antigens and pemphigus foliaceous antigens.

A flow cytometric method was developed to perform differential leukocyte counts on bovine blood. Blood specimens from 50 healthy Holstein cows were analyzed by use of a flow cytometer. The method entailed diluting blood with phosphate-buffered, hypotonic saline solution containing acridine orange, and performing a step-wise, 3-parameter analysis on the bases of cell size, cellular granularity, and granulocyte fluorescence. Initially, proportions of monocytes, granulocytes, and lymphocytes were determined by creating appropriate windows on dot plots of cell size (determined by forward light scatter) vs cellular granularity (determined by the logarithm of side light scatter). Eosinophils were resolved by analysis of granulocytes as dot plots of logarithms of green vs red fluorescence ascribed to acridine orange. Proportions of eosinophils and neutrophils were computed from data so generated. Microclumps of platelets spuriously affected counts of some granulocytes, particularly eosinophils. Differential leukocyte counts determined by flow cytometry generally compared favorably with those obtained by use of the conventional microscopic method, using Wright-stained blood films. Mean neutrophil and eosinophil counts determined by the 2 methods did not differ significantly, but lymphocyte counts determined by flow cytometry were significantly higher than those determined by microscopy (P < 0.01). Correlation coefficients for counts of neutrophils, eosinophils, and lymphocytes determined by the 2 methods ranged from 0.519 to 0.833. Correlation between monocyte counts was low (r = 0.147), although mean monocyte counts determined by the 2 methods did not differ significantly. Total leukocyte counts determined by flow cytometry were significantly lower (P < 0.01) than counts determined by use of an automated cell counter; correlation between the 2 counts was low (r = 0.350).

Pneumonic pasteurellosis was experimentally induced in calves by inoculation of 5 X 10(8) Pasteurella haemolytica organisms into the right diaphragmatic lung lobe. Blood and bronchoalveolar lavage fluid samples were obtained prior to inoculation and at postinoculation hour (PIH) 2, 4, and 6. Calves developed acute lung injury, characteristic of pneumonic pasteurellosis. Lesions were found only in the right diaphragmatic lobe. By PIH 4, significant (P < 0.01) increases were detected in lavage fluid total cell count, neutrophil count, total protein and albumin concentrations, and alkaline phosphatase (ALP) and lactic dehydrogenase (LD) activities. Myeloperoxidase and elastase activities did not increase. Neutrophil depletion ameliorated the lung lesions and prevented the increase in lavage fluid cell count, total protein, and albumin concentrations and ALP and LD activities. Treatment with the iron chelator, deferoxamine mesylatehydroxyethyl starch, attenuated the increase in total protein and albumin concentrations and ALP and LD activities at PID 4, but not PIH 6. Treatment with a neutrophil function inhibitor, pentoxifylline, prevented the increase in lavage fluid neutrophil numbers, but accentuated the increase in total protein and albumin concentrations, and ALP, LD, myeloperoxidase, and elastase activities.