Temporal changes, as well as differences in distribution, in concentrations of 24 amino acids in plasma and whole blood of neonatal foals were determined from birth to 2 days of age. In addition, differences in concentrations of amino acids in plasma between mare and foal pairs were determined at birth. Significant (P < 0.05) hypoaminoacidemia existed for 15 amino acids in plasma of foals at birth, compared with mares (paired t-test). Concentrations of 7 amino acids (aspartate, glutamate, glutamine, glycine, hydroxyproline, phenylalanine, proline) in plasma of foals were higher (P < 0.05) at birth than in mares, and concentrations of 2 (taurine, tryptophan) were not different (P > 0.05). Significant (P < 0.05) temporal changes for concentrations of 19 of 24 amino acids in plasma were observed during the 48-hour period. Concentrations of 13 of the 19 amino acids in plasma that had significant changes were higher (P < 0.05) at 48 hours. Significant (P > 0.05) effect of time on concentration of 5 amino acids (alanine, methionine, phenylalanine, taurine, threonine) in plasma was not found after birth. Temporal changes in concentrations of 7 amino acids (alanine, asparagine, glutamine, histidine, hydroxyproline, methionine, and threonine) in whole blood were not significantly (P > 0.05) different from those in plasma. Temporal changes for concentrations of the remaining 17 amino acids in whole blood were significantly (P < 0.05) different, compared with plasma. Distribution of the concentrations of 18 amino acids between whole blood and plasma was significantly (P < 0.05) different. Concentrations of 5 amino acids (citrulline, cystine, glutamine, methionine, tryptophan) were significantly (P < 0.05) lower in whole blood than in plasma, whereas concentrations of 13 amino acids were significantly (P < 0.05) higher in whole blood vs plasma. Concentrations of 6 amino acids (asparagine, isoleucine, leucine, proline, serine, valine) in whole blood were not significantly different from concentrations in plasma. Significant differences in temporal patterns of concentrations of amino acids in plasma and whole blood may be attributable to nutritional or physiologic changes associated with parturition. Significant differences between concentrations of amino acids in whole blood and plasma may be attributable to ontogeny or specificity of transport systems across cell membranes.

Dexmedetomidine (Dex), an alpha 2-receptor agonist, is the pharmacologically active d-isomer of medetomidine, a compound used as a sedative in veterinary medicine. Isoflurane anesthetic requirement (minimum alveolar concentration; MAC), rectal temperature, and cardiorespiratory variables were studied in chronically instrumented Yucatan miniature swine during DEX (20 micrograms/kg of body weight)-induced changes in body temperature. All studies were performed at room temperature of 22 C. The DEX was given as a 2-minute infusion into the left atrium. Each pig was studied twice. For protocol 1, the core temperature of the pigs was maintained at (mean +/- SD) 38.2 +/- 0.5 C by use of a thermostatically controlled water blanket and a heating lamp. For protocol 2, the core temperature was not externally manipulated and it decreased from 38.2 +/- 0.4 C to 32.2 +/- 1.2 C during the more than 3 hours of the protocol. Control isoflurane MAC was 1.66 +/- 0.2% and was 1.74 +/- 0.3% for protocols 1 and 2, respectively; DEX decreased MAC by 34 and 44%, respectively. For protocol 1, reduction in MAC after DEX administration returned by 50 and 80% at 84 and 138 minutes, respectively. If rectal temperature was not maintained (eg, allowed to decrease), MAC was reduced by 57% at the same time as the return to 80% in the swine with maintained body temperature. Respiratory rate and minute ventilation were significantly higher in swine with maintained temperature. The PaCO2 was lower and, accordingly, pH was higher in these swine. Blood pressure and heart rate were not affected by temperature changes.

Generation of free radicals and the ability of various antioxidants to attenuate radical production in freshly procured cancellous bone specimens was investigated, using spin-trapping and electron spin resonance (ESR) techniques. Seven core cancellous bone specimens, 10 mm long and 7.9 mm in diameter, were obtained using aseptic technique, from the proximal portion of the humerus of 9 adult mixed-breed dogs. One core cancellous bone specimen from each dog was incubated in spin trap alpha-phenyl-N-tert-butylnitrone in Eagle's minimum essential medium and served as a control. The other 6 specimens from each dog were incubated in alpha-phenyl-N-tert-butylnitrone/Eagle's minimum essential medium plus 1 of the following antioxidants: superoxide dismutase, catalase, superoxide dismutase/catalase, indomethacin, allopurinol, or deferoxamine mesylate. All specimens were incubated at 26 C for 90 minutes, then frozen at -20 C until they were prepared for analysis by ESR spectroscopy. Each specimen was thawed, homogenized, and extracted in a low-dielectric organic solvent prior to obtaining an ESR spectrum which was analyzed for hyperfine splitting constants to identify radicals. Each first-derivative spectrum was digitally double-integrated to obtain an area: these areas were used to compare intensities of the spin. For each treatment group, the areas from the treated specimens were compared with the areas from the control specimens, using a paired t-test. Significance was accepted at P less than or equal to 0.05. Spin adducts were detected in all cancellous bone specimens. Specimens incubated in deferoxamine (P = 0.0017) and superoxide dismutase/catalase (P = 0.0452) had significantly smaller areas than did control specimens. The areas for the other treatment groups did not differ significantly from controls. Our results substantiate free radical production in freshly procured cancellous bone specimens and that radical formation is attenuated by in vitro incubation with deferoxamine or superoxide dismutase/catalase.

Bronchoalveolar lavage (BAL) was performed on 16 horses to determine whether it caused local or diffuse inflammation in the lungs. In 7 horses, BAL was performed in both lungs twice, 48 hours apart. Although total cell counts of the BAL samples did not change significantly, there were increased numbers and percentage of neutrophils in the second lavage fluid samples. In 5 horses, BAL was performed in 1 lung and repeated 48 hours later in the same lung and in the corresponding airway in the contralateral lung. The absolute cell count and percentage of neutrophils were significantly (P = < 0.05) increased in the second sample from the lung that was lavaged twice. In 4 horses, BAL was performed in 1 lung and 48 hours later, repeated in an adjacent airway to the first BAL site, and in the corresponding airway in the contralateral lung. Significant differences were not detected in the total or differential cell counts of the BAL fluid recovered at any time, except for an increase in neutrophil percentage in the second sample in the contralateral lung. The increased neutrophil percentage values were within the range of normal for healthy horses. Results of the study suggested that, in horses, BAL induces a localized pulmonary neutrophil influx that persists at least 48 hours and is characterized by a relative and absolute increase in the number of neutrophils in the lavage fluids.

We used monoclonal antibodies and immunohistologic examination of lymph nodes, to elucidate the pathogenesis of lymphosarcoma induced by, infection with bovine leukemia virus (BLV). The superficial cervical lymph nodes from 3 BLV-infected but apparently healthy sheep and 5 sheep with full-blown lymphosarcoma were examined. We also investigated the integration of bovine leukemia provirus by use of Southern blotting. In lymph nodes from sheep lacking clinical signs of infection, in which the provirus had been integrated at multiple sites in the genome, many large hypertrophic follicles were observed in the cortex. These follicles had germinal centers consisting of CD4+T cells and B cells that expressed surface IgM (sIgM) and major histocompatibility complex (MHC) class-II antigens, but not B cell-specific B2 molecule. The percentage of CD4+T cells in the cortex was significantly (P < 0.05) higher than that of the controls and sheep with lymphosarcoma. In all sheep with lymphosarcoma, the lymph nodes were completely destroyed by proliferating neoplastic cells, and in addition, small atrophic follicles, which consisted of normal B-cell marker-positive cells, were seen near the trabecula and the subcapsule. In these instances, neoplastic cells appeared to be a monoclonal population derived from a single CD5- B-cell lineage and to be classified as 2 types, CD5-CD4-CD8-B2+MHC class-II+sIgM+ and CD5-CD4-CD8-B2+MHC class-II+sIgM-. Moreover, CD8+T cells infiltrated diffusely throughout the tumorous lymph nodes apart from the atrophic follicles, and CD4+T cells were observed around atrophic follicles. Both types of T cells were small-size, normal lymphocytes with round and noncleaved nuclei, and were apparently non-neoplastic cells. In fact, after separation by use of a panning method, it seems that, in blood mononuclear cells from BLV-infected sheep without clinical signs of infection, but in lymphosarcomatous stages, the proviral genome was integrated only in B cells and not in T cells. Thus, we conclude that the host's immune response may be still maintained at a lymphosarcomatous stage.

To differentiate Mycoplasma hyopneumoniae, the cause of mycoplasmal pneumonia in pigs, from M flocculare and M hyorbinis, an assay, using the polymerase chain reaction to amplify a segment of the 16S rRNA gene sequence, was developed. The assay was found to be useful for identification of field isolates, as well as for identification of laboratory-adapted strains. Amplification of DNA from M hyopneumoniae and M flocculare resulted in products of 200 and 400 base pairs, respectively. The DNA from M hyorbinis was not amplified. The assay was sensitive enough to detect as little as 1,000 genome equivalents of M hyopneumoniae and M flocculare DNA. Sensitivity was increased 100-fold by increasing the concentration of magnesium ion in the reaction buffer from 2 to 4 mM; however, DNA from M hyorbinis was also amplified under these conditions. The DNA from several walled bacteria and from other mycoplasmas was also tested, but none of these DNA samples was amplified, suggesting that the assay was specific for porcine mycoplasmas.

Pharmacologically induced splenic contraction might be useful during certain medical or surgical procedures in horses. The effects of phenylephrine, an alpha 1-adrenergic receptor agonist, on hemodynamic function and splenic dimensions were examined in 6 healthy adult horses. Phenylephrine infusion (1, 3, or 6 microgram/kg of body weight/min for 15 minutes) resulted in a dose-related increase in mean pulmonary artery pressure; right atrial pressure; systolic, mean, and diastolic arterial pressures; and packed cell volume (P = 0.0001). Concurrent decreases in heart rate and specific cardiac output (P = 0.0001) were detected, but stroke volume did not vary significantly. The rate-pressure product was increased only at the highest phenylephrine dosage (P = 0.012). Bradycardia was observed at all dosages during drug infusion, and second-degree atrioventricular block was detected in 88% of horses during infusion. Phenylephrine administration caused dose-dependent splenic contraction, as detected by ultrasonographic measurements of splenic area and thickness (P = 0.0001). At the 3- and 6-microgram/kg/min infusion rates, splenic area was reduced to 28 and 17% of baseline measurement, respectively. Splenic dimensions had returned to baseline values by 35 minutes after the end of infusion. Infusion of phenylephrine at a dosage of 3 microgram/kg/min for 15 minutes can be used to induce splenic contraction in horses.

Growth of Pasteurella haemolytica A1 in RPMI 1640 medium containing 0.5% bovine serum albumin (BSA) for 2.5 hours enhanced culture supernatant leukotoxic activity [30,700 +/- 12,900 toxic units/ml, compared with leukotoxic activity of culture supernatants produced in RPMI 1640 medium alone (120 +/- 40 toxic units/ml)]. Gel filtration chromatography of the leukotoxic activity from RPMI 1640 medium supernatants in buffer containing 50 mM NaCl indicated a single leukotoxic activity peak (peak I) eluting near the gel resin molecular mass exclusion limit (estimated molecular mass of approx 8,000 kd). In contrast, culture supernatants produced in RPMI 1640 plus bovine serum albumin medium (RPMI + BSA) had peak I and 2 additional leukotoxic activity peaks (peaks II and III) with estimated molecular mass of approximately 80 and < 30 kd, respectively. All leukotoxic activity peaks were composed of approximately 100-kd molecular mass leukotoxin protomer, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with a monoclonal antibody against leukotoxin. Subjecting culture supernatant leukotoxic activity produced in RPMI + BSA to gel filtration chromatography in buffer containing 500 mM NaCl or 6M urea resulted in detection of only a single leukotoxic activity peak with estimated approximate molecular mass of 250 and 800 kd, respectively. These findings suggest that P haemolytica exists as a high molecular mass aggregate with low leukotoxic activity which, in the presence of BSA, partially disaggregates to multiple toxin forms with enhanced leukotoxic activity. Some of these leukotoxin forms interact with dextran-based gel resins at low ionic strength.

Effects of 2 drugs commonly used for chemical restraint of cattle were evaluated for their effect on laryngeal and pharyngeal anatomy, function, and response to stimuli. Eighteen adult Jersey cows, free of respiratory tract disease, were studied. Cows were assigned at random to 1 of 3 treatment groups. Endoscopic evaluations were performed before and at a predetermined time interval after administration of each drug. Responses to stimuli were evaluated by stimulating 7 preselected sites (epiglottis, left and right arytenoid cartilages, left and right vocal folds, and left and right dorsolateral pharyngeal walls) with a closed, transendoscopic biopsy probe. Xylazine HCl (0.05 mg/kg of body weight, IV) was administered to group-1 cows (n = 6), and endoscopy was repeated 5 minutes after administration of the drug. Xylazine (0.07 mg/kg, IV) was administered to group-2 cows (n = 6), and endoscopy was repeated 5 minutes after administration of the drug. Acepromazine maleate (0.035 mg/kg, iv) was administered to group-3 cows (n = 6), and endoscopy was repeated 10 minutes after administration of the drug. Responses to stimuli were scored as brisk (0), moderate (1), slow (2), and absent (3). Scores for responses to stimuli were compared, using the Wilcoxon signed-rank test for data within groups, and a general linear models procedure, using the Kruskal-Wallis test between groups. Interobserver agreement rates were generated for each group. A value of P<0.05 was considered significant. Xylazine profoundly changed laryngeal sensitivity and function at both dosages. The corniculate processes of the arytenoid cartilages were observed to be in a markedly adducted position after sedation. Response to stimuli was significantly (P = 0.03) slower than normal after sedation, using both dosages. Displacement of the soft palate dorsal to the epiglottis was persistent in 50% of the cows after stimulation tests subsequent to sedation with xylazine. Acepromazine had a mild effect on laryngeal sensitivity and function. The corniculate processes of the arytenoid cartilages were observed in paramedian position after sedation. Acepromazine did not significantly affect responses to stimuli. Effects of sedation on responses to stimuli were not significantly different for groups 1 and 2. However, effects for group 3 were significantly different from those for groups 1 and 2 (P = 0.006 and 0.004, respectively). Endoscopic evaluation of the proximal portion of the respiratory tract of cattle should be performed without sedation, when possible. If sedation is required to facilitate restraint for endoscopy, acepromazine maleate is recommended over xylazine on the basis of results of this study.

A commercial rapid-absorbed ELISA developed to detect antibodies to Mycobacterium paratuberculosis in bovine serum was modified for use with goat serum. Diagnostic sensitivity was evaluated, using a group of 163 goats from a herd with endemic paratuberculosis. Blood and fecal samples were obtained simultaneously, and prevalence of shedding of M paratuberculosis in the feces was estimated by detection of DNA of the mycobacterial insertion sequence, IS900, using a commercial test kit. Diagnostic specificity was evaluated, using blood samples from a total of 123 goats in 10 herds that were considered clinically free of paratuberculosis. The IS900 DNA was detected in 35 of the 163 goats (21%) from the infected herd. Serum antibody to M paratuberculosis was detected in 19 of the 35 IS900 DNA-positive goats, for apparent sensitivity of 54%. Serum antibody was detected in 18 of the 128 IS900 DNA-negative goats from the infected herd. Negative results for serum antibody to M paratuberculosis were obtained for all 123 goats from the herds that were considered clinically free of paratuberculosis.