Objective-To evaluate the effect of controlled exposure to inhaled lipopolysaccharides (LPS) on the pulmonary inflammatory response of anesthetized pigs. Animals-Forty-seven 8- to 12-week-old domestic pigs. Procedure-Pigs were anesthetized with pentobarbital, instrumented for measurement of cardiopulmonary function, and randomly assigned to receive saline (0.9% NaCl) solution or 0.25, 0.5, or 1.0 µg of LPS/kg/h for 2 or 6 hours via nebulization through the endotracheal tube. Cardiopulmonary variables were measured, ex vivo neutrophil superoxide production determined, and postmortem assessment for pulmonary neutrophil influx and modulation of adhesion molecule (E-selectin) expression was done. Results-Mild changes in cardiopulmonary function were observed in response to inhaled LPS in the 2- and 6-hour groups. In pigs inhaling LPS (0.5 or 1.0 µg/kg/h) for 6 hours, there was significant pulmonary neutrophil influx observed postmortem. An increase in expression of E-selectin on pulmonary endothelial cells after 6 hours of LPS inhalation (0.5 µg/kg/h) was also observed. In contrast, there was no significant influx of neutrophils or expression of E-selectin in lungs from pigs inhaling LPS for 2 hours. Conclusion and Clinical Relevance—Inhalation of LPS resulted in localized pulmonary inflammation characterized by neutrophil influx and increased expression of the endothelial cell adhesion molecule, E-selectin. It may be possible to relate our experimental findings to the clinical consequences of airborne LPS exposure in swine confinement facilities.

Objective-To determine the effect of caprine arthritis-encephalitis virus (CAEV) infection on expression of interleukin-16 (IL-16). Animals-6 goats experimentally infected with CAEV and 6 age-matched healthy uninfected control goats. Procedure-Peripheral blood mononuclear cells (PBMCs) and synovial membrane cells from infected and control goats cultured with or without phytohemagglutinin were analyzed for IL-16 mRNA by use of a reverse transcriptase-polymerase chain reaction assay with goat-specific primers, after cloning and sequencing of a 384-bp fragment of the goat IL-16 gene. Synovial fluid, serum, and culture supernatants of PBMCs and synovial cells of control and CAEV-infected goats were analyzed for IL-16 by use of an ELISA. Results-The 384-bp product was 86% homologous to the corresponding human IL-16 nucleotide sequence. Higher expression of IL-16 mRNA in PBMCs (unstimulated or stimulated with phytohemagglutinin) was detected in samples from CAEV-infected goats, compared with control goats, but the difference was not significant. Synovial membrane cells infected in vitro had higher expression than uninfected control cells. Higher IL-16 concentration was detected in synovial fluid, serum, and culture supernatants of PBMCs of infected goats than in samples from control goats. Conclusions and Clinical Relevance-Infection with CAEV increases expression of IL-16, a proinflammatory and chemotactic cytokine. This cytokine appears to be constitutively expressed at low concentrations in normal uninfected PBMCs and synovial membrane cells. Increased production of IL-16 in CAEV infection may partly be responsible for increased lymphoid cell infiltrations observed in arthritic joints and other tissues of CAEV-infected goats.

Objective-To purify and partially characterize various isoforms of canine pepsinogen (PG) from gastric mucosa. Sample Population-Stomachs obtained from 6 euthanatized dogs. Procedure-Mucosa was scraped from canine stomachs, and a crude mucosal extract was prepared and further purified by use of weak anion-exchange chromatography, hydroxyapatite chromatography, size exclusion chromatography, and strong anionexchange chromatography. Pepsinogens were characterized by estimation of molecular weights, estimation of their isoelectric points (IEPs), and N-terminal amino acid sequencing. Results-Two different groups of canine PG were identified after the final strong anion-exchange chromatography: PG A and PG B. Pepsinogens differed in their molecular weights and IEP. Pepsinogen B appeared to be a dimer with a molecular weight of approximately 34,100 and an IEP of 4.9. Pepsinogen A separated into several isoforms. Molecular weights for the various isoforms of PG A ranged from 34,200 to 42,100, and their IEPs ranged from 4.0 to < 3.0. The N-terminal amino acid sequence for the first 25 amino acid residues for PG A and B had good homology with the amino acid sequences for these proteins in other species. Conclusions and Clinical Relevance-Canine PG B and several isoforms of canine PG A have been purified. Availability of these PGs will facilitate development of immunoassays to measure PG in canine serum as a potential diagnostic marker for gastric disorders in dogs.

Objectives-To evoke and measure the nociceptive withdrawal reflex (NWR) by use of electromyographic recordings and to score the behavioral nociceptive responses to electrical pulses in standing nonsedated horses. Animals-10 adult horses. Procedure-The lateral palmar digital nerve of the forelimb was transcutaneously stimulated, and surface electromyographic responses were recorded from the ulnaris lateralis, extensor carpi radialis, and common digital extensor muscles. Stimuli consisted of a 25-millisecond train of 5 constant-current pulses delivered by a computer-controlled stimulator. The 80- to 250-milliseconds poststimulation interval was analyzed to detect the NWR. The current intensity was increased in steps of 0.5 mA until the NWR threshold intensity (It) was reached. The stimulus at It was repeated twice. Latency and amplitude of the NWR, together with the behavioral reaction of horses, were analyzed. The latter was scored according to a scale from 0 (no reaction) to 5 (vigorous reaction). Finally, 3 suprathreshold stimuli at 1.2 × It were analyzed. Results-The median It to elicit NWR was 2.5 mA. Median onset latency of the NWR was 96.0 milliseconds at It and 89.6 milliseconds for suprathreshold stimuli. The amplitude of the reflexes was higher for suprathreshold stimulations, and behavioral reactions were slightly stronger when stimulus intensity increased. Conclusions and Clinical Relevance-Results of our study indicate that it is possible to record NWR in conscious standing horses, to define a reflex threshold, and to measure reflexes in response to increasing stimulus intensity.

Objective-To evaluate the effect of 2 oxytocin products administered to sows at the onset of fetal expulsion on the integrity of umbilical cords, meconium staining, and piglet mortality. Animals-2099 neonatal pigs. Procedure-180 parturient sows were randomly assigned to 3 stratified groups of 60 sows each. Two groups of sows were injected IM at the onset of fetal expulsion with 1 of 2 oxytocin commercial products (20, 40, or 50 U for sows weighing 120 to 150 kg, 151 to 250 kg, or ≥ 251 kg, respectively). Control sows were treated IM with saline (0.9% NaCl) solution. Farrowing time, expulsion intervals, and numbers of stillborn and liveborn piglets were recorded for each sow. Piglets were evaluated for inspiratory effort, heart rates, and degree of meconium staining of skin (nonstained, and moderately or severely stained). Umbilical cords were classified as normal in appearance, edematous, congested, hemorrhagic, or ruptured. Results-Oxytocin-treated sows had a significant decrease in farrowing time and expulsion intervals and also had a significantly higher number of stillborn piglets per litter, compared with control sows. The number of piglets per litter with ruptured and hemorrhagic umbilical cords was significantly greater in oxytocin- treated sows, compared with control sows. In near-death stillborn piglets, oxytocin treatment significantly decreased inspiratory efforts at birth and increased the rate and severity of meconium staining, compared with saline treatment. Conclusions and Clinical Relevance-Oxytocin given to sows at the onset of fetal expulsion significantly increases the rate of fetal distress, anoxia, and intrapartum death in piglets.

Objective-To determine the ratio of ventral-to-dorsal transverse diameters between the wings of the sacrum on ventrodorsal radiographic views of the pelvis in large dogs and to validate the reliability of this morphometric analysis for functional interpretation. Sample Population-Pelvic specimens from 40 large-breed dogs and radiographs of 113 large-breed dogs. Procedure-In an anatomic and radiographic evaluation, the transverse dorsal diameter (TVDS) and transverse ventral diameter (TVV) between the wings of the sacrum were evaluated in sacrum specimens and on corresponding radiographs of the pelvis and sacrum. The ratio between TVV and TVDS (VD ratio) was calculated. Intraobserver reliability was determined by calculation of the coefficient of variation. In a retrospective radiographic evaluation, the VD ratio was determined in Rottweilers, Golden Retrievers, and German Shepherd Dogs. Correlations between VD ratio and breed, age, and sex were tested. Results-The VD ratio was significantly higher in Rottweilers than in Golden Retrievers and German Shepherd Dogs, denoting an oblique alignment of the sacral wings in Rottweilers (ie, the dorsal aspects of the sacral wings were located more medially than the ventral aspects) and an almost sagittal alignment in the other breeds. The VD ratio was significantly associated with age but not with sex. Conclusion and Clinical Relevance-Sagittal alignment of the wings of the sacrum is considered to be biomechanically less efficient. These results provide a basis for further studies to evaluate radiographic assessment of the sacroiliac joints similar to the evaluation for hip dysplasia.

Objective-To compare cardiopulmonary responses during anesthesia maintained with halothane and responses during anesthesia maintained by use of a total intravenous anesthetic (TIVA) regimen in horses. Animals-7 healthy adult horses (1 female, 6 geldings). Procedure-Each horse was anesthetized twice. Romifidine was administered IV, and anesthesia was induced by IV administration of ketamine. Anesthesia was maintained for 75 minutes by administration of halothane (HA) or IV infusion of romifidine, guaifenesin, and ketamine (TIVA). The order for TIVA or HA was randomized. Cardiopulmonary variables were measured 40, 60, and 75 minutes after the start of HA or TIVA. Results-Systolic, diastolic, and mean carotid arterial pressures, velocity time integral, and peak acceleration of aortic blood flow were greater, and systolic, diastolic, and mean pulmonary arterial pressure were lower at all time points for TIVA than for HA. Pre-ejection period was shorter and ejection time was longer for TIVA than for HA. Heart rate was greater for HA at 60 minutes. Minute ventilation and alveolar ventilation were greater and inspiratory time was longer for TIVA than for HA at 75 minutes. The PaCO2 was higher at 60 and 75 minutes for HA than for TIVA. Conclusions and Clinical relevance-Horses receiving a constant-rate infusion of romifidine, guaifenesin, and ketamine maintained higher arterial blood pressures than when they were administered HA. There was some indication that left ventricular function may be better during TIVA, but influences of preload and afterload on measured variables could account for some of these differences.

Objective-To compare experimentally induced concurrent infection with bovine viral diarrhea virus (BVDV) and bovine rotavirus (BRV) with infection of either virus alone in calves. Animals-Seventeen 1-day-old gnotobiotic calves. Procedure-Calves were allotted to 8 treatments as follows: group 1, mock-infected control calves (n = 2); group 2, inoculated with BVDV on day 1 (2); groups 3, 5, and 7, inoculated with BRV on days 1 (2), 4 (1), or 7 (2), respectively; and groups 4, 6, and 8, inoculated with BVDV on day 1 and with BRV on days 1 (2), 4 (2), or 7 (4), respectively. Concentrations of BVDV in serum and ileal tissues were measured, and BRV shedding in feces was determined. Histologic examination and immunohistochemical analysis were conducted to detect lesions and viral antigens. Results-Neonatal calves inoculated with BVDV alone or with BVDV on day 1 and BRV on day 7 developed villus atrophy and submucosal inflammation of the intestines. Concurrent BVDV and BRV infections acted synergistically in the intestinal tract, causing more severe enteric disease than infection with either virus alone. Severe lymphoid depletion was associated with BVDV infection in calves regardlesss of concurrent BRV infection. Conclusions and Clinical Relevance-Infection with BVDV played direct and indirect roles in enteritis in neonatal calves, causing villus atrophy in the duodenum and submucosal inflammation of the intestines. Also, BVDV potentiated effects of BRV. Concurrent infection with BVDV and BRV resulted in more severe enteric disease in neonatal calves than infection with BRV or BVDV alone.

Objective-To determine the prevalence of activating internal tandem duplications (ITDs) in exons 11 and 12 of c-kit in mast cell tumors (MCTs) of dogs and to correlate these mutations with prognosis. Sample Population-157 formalin-fixed, paraffinembedded MCTs from dogs in the pathology database of the Veterinary Medical Teaching Hospital at the University of California, Davis. Procedure-Genomic DNA was isolated from tumor specimens and a polymerase chain reaction procedure was performed to determine whether there were ITDs in exons 11 and 12. Results-We identified ITDs in 1 of 12 (8%) grade-I, 42 of 119 (35%) grade-II, and 9 of 26 (35%) grade-III tumors (overall prevalence, 52 of 157 [33%]). Logistic regression analysis revealed that the odds of grade-II and -III tumors possessing an ITD were approximately 5 times greater than that for grade-I tumors, although these odds did not differ significantly. Although MCTs possessing an ITD were twice as likely to recur after excision and twice as likely to result in metastasis as those without an ITD, these values also did not differ significantly. Conclusions and Clinical Relevance-These results provide evidence that ITDs in c-kit occur frequently in MCTs of dogs. The high prevalence of c-kit activating mutations in MCTs of dogs combined with the relative abundance of mast cell disease in dogs provide an ideal naturally developing tumor in which to test the safety and efficacy of novel small-molecule kinase inhibitors such as imatinib mesylate.

Objective-To determine the effect of experimental intraluminal distention on microvascular perfusion of the small colon in horses. Animals-6 mixed-breed healthy horses (mean age [+/- SD], 9.1 +/- 2 years). Procedure-Under general anesthesia, the small colon was exposed by celiotomy and 3 segments were demarcated. In 1 of these segments, intraluminal obstruction was created by placement of a latex balloon inflated to a pressure of 40 mm Hg (obstructed segment). The other segments were the sham-operated segment and the control segment. Microvascular perfusion was evaluated in the mucosal, submucosal, muscular, and serosal layers by injection of 15--µm-diameter colored microspheres into branches of the caudal mesenteric artery. Recovery of microspheres was performed by tissue digestion, washing, and centrifugation. Distribution of microspheres in the intestinal layers was evaluated by direct observation of stained frozen sections by light microscopy. Results-A significant reduction was observed in total microvascular perfusion of obstructed segments, which was 26.4% of that of control segments. This reduction was not evident in the mucosal layer. Conclusion and Clinical Relevance-Intraluminal distention of the equine small colon wall can promote ischemia by a reduction in microvascular perfusion in the intestinal wall. Intestinal layers do not seem to be affected to the same extent, because the absolute value for mucosal perfusion did not decrease in the obstructed segment.