Alterations in the size and functions of porcine alveolar macrophages exposed to sublytic amounts of heat-labile, hemolytic cytotoxin produced by Actinobacillus pleuropneumoniae (App) serotype 1, strain HS54 into the culture medium were studied in vitro. Alveolar macrophages were sensitive to the cytotoxin; treatment of the macrophages with low concentrations of cytotoxin (0.016 hemolytic unit) resulted in severe, irreversible cell swelling. However, high doses of cytotoxin (2.0 hemolytic units) were required to cause substantial cell death, as indicated by the influx of propidium iodide into and release of lactate dehydrogenase from cells. Macrophages exposed to low, sublytic doses of cytotoxin failed to migrate toward chemoattractant, were unable to attach to glass, and failed to phagocytize optimally opsonized erythrocytes. Macrophages already attached to glass surfaces detached when exposed to sublytic doses of cytotoxin. The swelling and impairment of functions of alveolar macrophages observed in this study could not be attributed to endotoxic effects, because heat treatment of the cytotoxin preparation for 60 minutes at 60 C resulted in complete loss of cytotoxicity. We conclude that sublytic doses of heat-labile, hemolytic cytotoxic substances produced by App depress alveolar macrophage function at concentrations likely to develop in association with acute pulmonary infection with App. The Apx (A pleuropneumoniae Rtx toxins) exotoxins secreted by the bacteria into culture medium were considered responsible for the toxic activity of the cytotoxin preparation. The Apx of the App field strain used in this study were likely to be similar to those of serotype-1 reference strain (S4707). Analysis by use of DNA-DNA hybridization indicated that genomic DNA of the field strain contained sequences similar to those encoding structural protein of ApxI (apxIA) and ApxII (apxIIA) of the serotype-1 reference strain. Therefore, Apx produced by the field strain of App used in this study are likely to be of similar pathogenic importance worldwide.

Force platform analysis of gait provides ground reaction force information that can be used to study limbs with normal or abnormal function. When combined, the interrelated variables of ground reaction forces give a more thorough description of gait than when used individually. To describe the pattern of ground reaction forces in clinically normal, conditioned, mesomorphic dogs, we studied the data from platform gait analyses of 43 dogs. Mediolateral (Fx), craniocaudal (Fy), and vertical (Fz) forces were measured and recorded. Torque (Tz) around the vertical axis also was calculated. Mean stance times for forelimbs and hind limbs were 0.278 and 0.261 second, respectively. Among dogs, ground reaction forces were normalized and expressed as percentage of body weight (%bw). The vertical (Fz) peak, average force during stance phase, and force vs time impulses were 106.68, 60.82, and 17.2 %bw in forelimbs, and were 65.11, 35.3, and 9.33 %bw in hind limbs. The forelimb braking/ propulsive (Fy) peaks were -16.74 and +6,73 %bw. In hind limbs, these peaks were -3.76 and +7.69 %bw. The usual mediolateral force (Fx) pattern found in forelimbs was laterally directed, with average peak magnitude of 6.69 %bw, whereas the hind limb patterns were variable.

Although healthy and diseased bovine respiratory tracts have been intensively studied during the last years, to the authors' knowledge, there have been no attempts to objectively examine the inspiratory drive from the brain to the nerves and muscles and its transformation in pressure. Such technique would be useful in assessing the possibility of altered ventilatory drive or inspiratory muscle fatigue in the context of an animal with ventilatory failure. The relation among ventilation, airway opening occlusion pressure generated 100 milliseconds after onset of inspiration (Pawo100ms) and 6 indexes describing diaphragmatic electromyographic activity (EMGdi) recorded via implanted fishhooks was evaluated during free and impeded CO2, rebreathing in 6 young bulls. The best significant linear correlations (r > 0.8) with inspiratory center afferent stimulation, as judged by end-tidal CO2 concentration in expired air, were found for Pawo100ms, peak moving time average or variance EMGdi, and mean integrated EMGdi, whatever had been the respiratory impedance. However, with an inspiratory load, Pawo100ms responses systematically had greater increase for a given change in the driving EMGdi, implying dependence of the former not only on neural input, but also on configurational factors that determine inspiratory muscle excitation-pressure generation couplings. The reproducibility of EMGdi absolute values and changes was satisfactory up to 10 hours, but could not be repeated from one day to the other. It was concluded that, provided the constancy of the electrical coupling of the recording system to the tissue being studied is ensured, specific EMGdi and Pawo100ms values correlate reliably with amount of CO2 during free and loaded breathing. Simultaneous collection of both values during experimentally induced pulmonary disease in calves could, therefore, produce information to help answer questions about the role of CNS and inspiratory muscle dysfunction in case of ventilatory failure. Careful interpretation, however, requires additional measurements, such as end-expiratory lung volume, and some familiarity with the underlying physiologic processes that link phrenic nerve discharge to generation of negative pressure at the airway opening.

An ELISA was developed to detect antibodies to the 41-kd flagellin (P41) of Borrelia burgdorferi in serum obtained from cattle. Absorption studies, immunoblot analysis, immunoelectron microscopy, and correlation of results of the P41-ELISA and the P39-ELISA as well as measurement of the antibody to P41 in calves challenge-exposed with Borrelia theileri were used to assess the specificity of the P41-ELISA. Antigens derived from Escherichia coli, Leptospira interrogans serovar hardjo, and B burgdorferi were used for absorption studies and immunoblot analysis. Antibodies to P41 of B burgdorferi cross-reacted with antigens of E coli, but were not cross-reactive with L hardjo. A value 3 SD higher than the mean of the negative-control population of cattle was defined as the minimum value cutoff value) for a positive result by the P41-ELISA. Use of this value for classification of test results reduced the predicted rate of false-positive results attributable to E coli cross-reactivity to 1%. Immunoblot analysis revealed that test-positive serum from cattle reacted mainly with 41-, 39-, 34-, and 31-kd proteins of B burgdorferi, as well as several smaller proteins. Immunoelectron microscopy revealed that serum from cattle that was test-positive by the P41-ELISA bound to the flagellin and outer membrane of B burgdorferi. Results of absorption studies, immunoblot analysis, and immunoelectron microscopy were correlated and indicated that serum from cattle that was test-positive by P41-ELISA had stronger reactivity to B burgdorferi antigens than to antigens of E coli or L hardjo. The concentrations of antibodies measured by P41-ELISA and P39-ELISA testing were highly correlated [R(2)=0.78]. Calves challenge-exposed with B theileri also had test-positive results by the P-41-ELISA as early as 2 weeks after exposure, but serum antibody concentrations decreased to prechallenge-exposure concentrations by 9 weeks after exposure. We concluded that the P41-ELISA was useful as a screening method to detect B burgdorferi infections in cattle.

Serum C-reactive protein (CRP) concentration was measured, using an automated immunoturbidimetric assay, in 44 clinically normal dogs and 67 dogs with band neutrophil count greater than or equal to 10(9) cells/L, and values were found to be significantly (P less than or equal to 0.05) different. Correlation of serum CRP concentration and band neutrophil count in the 67 dogs with greater than or equal to 10(9) band neutrophils/L resulted in a statistically significant P less than or equal to 0.05), but low correlation coefficient of 0.34. Serum CRP concentration and CBC values were determined for 6 clinically normal dogs undergoing anesthesia (controls) and 6 clinically normal dogs undergoing anesthesia and ovariohysterectomy. Significant alterations in CBC results and serum CRP concentration, compared with baseline values, were lacking in dogs of the control group. Serum CRP concentration was significantly (P less than or equal to 0.05) increased above baseline values in dogs undergoing surgery and was significantly (P less than or equal to 0.05) increased, compared with values in control dogs by 12 hours after surgery. In dogs undergoing surgery, serum CRP concentration was also significantly (P less than or equal to 0.05) different from values in control dogs at 28 and 36 hours, but not at the 76- and 124-hour sample collection times. Alterations in CBC values compatible with possible or convincing inflammation were detected in 83% of the dogs undergoing surgery at the 8- and 12-hour postsurgery sample collection times, 100% of dogs at 16, 22, 28, and 36 hours after surgery, 83% of dogs at 52 and 76 hours after surgery, 67% of dogs at 100 hours after surgery, and 0% of dogs at 124 hours after surgery. It was concluded that significant increases in CRP, concentration in dogs with surgical trauma were not detected earlier than CBC alterations compatible with possible or convincing inflammation.

To provide long-term gastric fistulas for collection of third-compartment gastric contents, Janeway mucosal tube gastrostomy was performed, using a gastrointestinal stapling instrument, in 6 castrated adult male llamas. Mean operative time (+/- SEM) was 65 +/- 4.16 minutes. All llamas survived the 6-week study period. Of the 6 llamas, 5 did not have signs of abdominal pain and returned to preoperative food consumption amounts within 36 hours. One llama had mild intermittent signs of abdominal pain daily for 7 days before returning to preoperative amount of food consumption. All gastrostomies leaked small amounts of gastric contents around indwelling 6- to 8-mm cannulas at the skin surface. Gastric contents did not leak when cannulas were dislodged from gastrostomy stomas. Replacement of cannulas was rapid and easy. Gravity-flow sample collection was best accomplished through 8-mm cannulas. Mean (+/- SEM) weight loss was detected in all llamas (15 +/- 3 kg) and was associated with frequent nonfeeding and stress of sample collection. Gross necropsy findings were unremarkable in 5 of 6 llamas. All mucosal tube gastrostomies were patent, and there was no evidence of peritonitis. One llama had a single fibrous adhesion connecting the operative site with the ascending colon. Histologically , small (2.5- to 15-mm diameter) partial-thickness mucosal erosions identified at the tube gastrostomy-gastric wall junctions may have been associated with indwelling gastric cannulas. The Janeway gastrostomy was generally well tolerated in the llamas and should be considered as a useful long-term fistulation technique.

Bovine immunoglobulin preparations from normal serum and from sera containing antibodies against Brucella abortus interfered with the brucellacidal action of bovine serum, whereas unfractionated normal serum and antisera were not inhibitory. The inhibitory property of immunoglobulin appeared to be attributable to some anticomplementary property because it also interfered with serum-mediated hemolysis of antibody-coated erythrocytes. The supernatant phase obtained after ultracentrifugation of bovine anti-B abortus immunoglobulin did not inhibit brucellacidal activity of normal bovine serum. Results of this study indicate that bovine anti-B abortus immunoglobulin preparations contain microaggregates of protein that can inhibit the ability of bovine serum to kill B abortus. The most likely mechanism is nonspecific activation of complement by microaggregated immunoglobulin, which consumes complement and makes it unavailable for bactericidal activity.

Influence of age, breed, and stage of pregnancy on hepatic ultrasonographic findings of cows was determined. In addition, the relation between body weight, height at the withers, milk production, and the measurements determined via ultrasonography was investigated. The liver of 186 cows was examined ultrasonographically. The cows comprised Swiss Braunvieh, Simmental, and Holstein breeds, and age ranged from 2.5 to 11.5 years. The ultrasonographic findings of the liver, gallbladder, caudal vena cava, and portal vein were described, and the position, size, thickness, and distal angle of the liver were determined. In addition, the position and diameter of the caudal vena cava and portal vein were determined. There was no significant difference between any of the variables determined and breed or age. Therefore, measurements for the 3 breeds and for the various ages were summarized into 1 group. There were significant correlations between body weight, milk production, and size and thickness of the liver. In 3 pregnant cows, the liver was examined ultrasonographically 8 times during the course of pregnancy. Positive correlation was detected between stage of pregnancy and diameter of the caudal vena cava. There was a negative correlation between stage of pregnancy and diameter of the portal vein. In 23 cows, the ultrasonographically determined measurements of the liver were compared with those determined at slaughter. Weight of the liver correlated well to thickness of the liver determined via ultrasonography.

Maximal conduction velocities of compound action potentials evoked by stimuli of 2 times threshold in the caudal cutaneous sural (CCSN) and medial cutaneous antebrachial (MCAN) nerves were determined by averaging potentials evoked and recorded through percutaneous needle electrodes. Mean maximal conduction velocities of compound action potentials were: CCSN = 61.3 +/- 2.0 meters/second (m/s) and MCAN = 56.4 +/- 2.8 m/s. To confirm accuracy of our percutaneous recordings, compound action potentials were recorded through bipolar chlorided silver electrodes from the exposed surfaces of fascicles of the CCSN and the MCAN. The maximal conduction velocities of these potentials were in agreement with the conduction velocities of compound action potentials that were evoked and recorded through percutaneous needle electrodes. The specificity of stimulating and recording sites was verified by recording before and after section of the nerves. Stimuli from 3 to 5 times threshold evoked a second, longer latency, compound action potential that consisted of a variable number of components in the CCSN and MCAN. The configurations and conduction velocities of the shorter latency potentials were the same as those of the single compound action potentials evoked by stimuli of 2 times threshold. Mean conduction velocities of the longer latency potentials were: CCSN = 24.4 +/- 2.6 m/s and MCAN = 24.5 +/- 2.2 m/s. Needle electrode and direct stimulation of either the CCSN or the MCAN at 3 to 5 times threshold failed to evoke contractions of limb muscles. Therefore, action potentials that contributed to the evoked compound potentials recorded in these horses arose, most likely, from afferent nerve fibers.

Polymyxin B and an antiserum against an Re mutant Salmonella typhimurium were evaluated for protective effect in an equine model of endotoxemia. Six 3- to 5-month-old foals were given endotoxin (0.25 micrograms/kg of body weight) IV after no pretreatment, or pretreatment with polymyxin B (6,000 U/kg, IV) or S typhimurium antiserum (1.5 ml/kg, IV). When given without pretreatment, endotoxin caused transient recumbency and increases in rectal temperature, and heart and respiratory rates. In addition, leukopenia and increases in circulating tumor necrosis factor (TNF) and interleukin 6 (IL-6) activities were detected. Compared with results obtained when endotoxin was given alone, pretreatment with polymyxin B resulted in significantly (P < 0.05) lower maximal plasma TNF and IL-6 activities, and significantly lower rectal temperature and respiratory rate. In contrast, compared with effects of endotoxin given without pretreatment, use of antiserum was associated with significantly (P < 0.05) higher respiratory rate, maximal plasma IL-6 activity, and total TNF response (as determined by areas under curves of plasma TNF vs time). These results indicate that polymyxin B may have potential as a treatment for equine endotoxemia. Salmonella typhimurium antiserum had no positive effect in this model, and, under certain conditions, may exacerbate the actions of endotoxin.