Objective—To determine whether canine tumor cell lines express functional tissue factor and shed tissue factor-containing microparticles. Sample—Cell lines derived from tumors of the canine mammary gland (CMT12 and CMT25), pancreas (P404), lung (BACA), prostate gland (Ace-1), bone (HMPOS, D-17, and OS2.4), and soft tissue (A72); from normal canine renal epithelium (MDCK); and from a malignant human mammary tumor (MDA-MB-231). Procedures—Tissue factor mRNA and antigen expression were evaluated in cells by use of canine-specific primers in a reverse transcriptase PCR assay and a rabbit polyclonal anti-human tissue factor antibody in flow cytometric and immunofluorescent microscopic assays, respectively. Tissue factor procoagulant activity on cell surfaces, in whole cell lysates, and in microparticle pellets was measured by use of an activated factor X-dependent chromogenic assay. Results—Canine tissue factor mRNA was identified in all canine tumor cells. All canine tumor cells expressed intracellular tissue factor; however, the HMPOS and D-17 osteosarcoma cells lacked surface tissue factor expression and activity. The highest tissue factor expression and activity were observed in canine mammary tumor cells and pulmonary carcinoma cells (BACA). These 3 tumors also shed tissue factor-bearing microparticles into tissue culture supernatants. Conclusions and Clinical Relevance—Tissue factor was constitutively highly expressed in canine tumor cell lines, particularly those derived from epithelial tumors. Because tumor-associated tissue factor can promote tumor growth and metastasis in human patients, high tissue factor expression could affect the in vivo biological behavior of these tumors in dogs.

Objective—To investigate the effects of twice-daily oral administration of hydrocortisone on the bile acids composition of gallbladder bile in dogs. Animals—6 placebo-treated control dogs and 6 hydrocortisone-treated dogs. Procedures—Dogs received hydrocortisone (median dose, 8.5 mg/kg) or a gelatin capsule (control group) orally every 12 hours for 84 days. Gallbladder bile samples were obtained via percutaneous ultrasound-guided cholecystocentesis from each dog before (day 0 [baseline]), during (days 28, 56, and 84), and after (days 28p, 56p, and 84p) treatment for differentiated quantification of unconjugated bile acids and taurine-conjugated and glycine-conjugated bile acids via high-performance liquid chromatography–tandem mass spectrometry. Results—Treatment with hydrocortisone for 84 days resulted in significant and reversible increases in the concentrations of unconjugated bile acids (ie, cholic, chenodeoxycholic, and deoxycholic acids) and a significant and reversible decrease in the concentration of total taurine-conjugated bile acids, compared with baseline or control group values. Treatment with hydrocortisone had no effect on bile concentrations of glycine-conjugated bile acids. Conclusions and Clinical Relevance—In dogs, hydrocortisone administration caused reversible shifts toward higher concentrations of the more hydrophobic unconjugated bile acids (chenodeoxycholic acid and deoxycholic acid) and toward lower concentrations of the amphipathic taurine-conjugated bile acids in gallbladder bile. These data suggest that similar bile acids changes could cause major alterations in gallbladder structure or function over time in hypercortisolemic dogs.

Objective—To investigate plasma disposition, concentration in the hair, and anthelmintic efficacy of eprinomectin after topical administration in donkeys. Animals—12 donkeys naturally infected with strongyle nematodes. Procedures—The pour-on formulation of eprinomectin approved for use in cattle was administered topically to donkeys at a dosage of 0.5 mg/kg. Heparinized blood samples and hair samples were collected at various times between 1 hour and 40 days after administration. Samples were analyzed via high-performance liquid chromatography with fluorescence detection. Fecal strongyle egg counts were performed by use of a modified McMaster technique before and at weekly intervals for 8 weeks after treatment. Results—Plasma concentration and systemic availability of eprinomectin were relatively higher in donkeys, compared with values reported for other animal species. Concerning the anthelmintic efficacy against strongyle nematodes, eprinomectin was completely effective (100%) on days 7 and 14 and highly effective (> 99%) until the end of the study at 56 days after treatment. No abnormal clinical signs or adverse reactions were observed for any donkeys after treatment. Conclusions and Clinical Relevance—Eprinomectin had excellent safety. The relatively high plasma concentration after topical administration could result in use of eprinomectin for the control and treatment of parasitic diseases in donkeys.

Objective—To evaluate a commercially available modified-live Streptococcus equi subsp equi vaccine for safety and persistence in vaccinated ponies and to detect recombination or reversion events in the vaccine strain. Animals—5 ponies that were 1.5 to 8 years old (group 1) and 4 ponies that were 6 months old (group 2). Procedures—Ponies were vaccinated, with a subsequent booster vaccination 2 to 3 weeks later, and monitored for 50 days. At booster vaccination, an equal amount of a tracycline-resistant wild-type strain of S equi was administered. Recovery of all strains was performed by use of bacteriologic culture and PCR assays. Results—Ponies in group 1 had background antibody titers against S equi antigen before vaccination despite the lack of known exposure to S equi. Ponies in group 2 were immunologically naïve. Increases in anti-S equi antibody titers were detected in both groups. Ponies in group 1 did not have clinical signs of disease caused by S equi. In group 2, all ponies developed abscesses in retropharyngeal lymph nodes; 1 pony developed severe clinical disease and was euthanized. The vaccine strain was recovered from ponies in group 2 for up to 24 days after vaccination. Conclusions and Clinical Significance—Although the vaccine was successful in inducing IgG antibodies against S equi in all ponies, findings suggested that the vaccine may have caused substantial morbidity and some deaths in the young ponies. In young ponies, the vaccine strain persisted in tissues for weeks; however, no evidence of recombination was detected.

Objective—To determine the influence of stifle joint flexion angle, cranial cruciate ligament (CrCL) integrity, tibial plateau leveling osteotomy (TPLO), and cranial tibial subluxation on the distance between the location of the origin and insertion of the CrCL (CrCLd) in dogs. Samples—4 pairs of pelvic limbs from adult dog cadavers weighing 23 to 34 kg. Procedures—Mediolateral projection radiographs of each stifle joint were obtained with the joint flexed at 90°, 105°, 120°, 135°, and 150°. Radiopaque markers were then placed at the sites of origin and insertion of the CrCL. Afterward, radiography was repeated in the same manner, before and after CrCL transection, with and without TPLO. Following CrCL transection, radiographs were obtained before and after inducing overt cranial tibial subluxation. Interobserver variation in measuring the CrCLd without fiduciary markers was assessed. The effect of CrCL integrity, cranial tibial subluxation, flexion angle, and TPLO on CrCLd was also determined. Results—Interobserver agreement was strong, with an intraclass correlation coefficient of 0.859. The CrCLd was significantly shorter (< 1 mm) at 90° of flexion; otherwise, flexion angle had no effect on CrCLd. Cranial tibial subluxation caused a 25% to 40% increase in CrCLd. No effect of TPLO on CrCLd was found, regardless of CrCL integrity, forced stifle joint subluxation, or flexion angle. Conclusions and Clinical Relevance—Overt cranial tibial subluxation in CrCL-deficient stifle joints can be detected on mediolateral projection radiographs by comparing CrCLd on neutral and stressed joint radiographs at joint angles between 105° and 150°, regardless of whether a TPLO has been performed.

Objective: To determine accuracy of the use of triaxial accelerometry for measuring daily activity as a predictor of maintenance energy requirement (MER) in healthy adult Labrador Retrievers. Animals—10 healthy adult Labrador Retrievers. Procedures: Dogs wore an accelerometer for two 2-week periods, with data on daily activity successfully collected for 24 to 26 days. These data, along with body weight, were used as independent variables in a multiple linear regression model to predict the dependent variable of daily MER. The predictive accuracy of the model was compared with that of a model that excluded activity. Dietary energy intake at a stated amount of body weight stability was used as an equivalent measure of MER in these analyses. Results: The multiple linear regression model that included body weight and daily activity as independent variables could be used to predict observed MER with a mean absolute error of 63.5 kcal and an SE of estimation of 94.3 kcal. Removing activity from the model reduced the predictive accuracy to a mean absolute error of 129.8 kcal and an SE of estimation of 165.4 kcal. Conclusions and Clinical Relevance: Use of triaxial accelerometers to provide an independent variable of daily activity yielded a marked improvement in predictive accuracy of the regression model, compared with that for a model that used only body weight. Improved accuracy in estimations of MER could be made for each dog if an accelerometer was used to record its daily activity.

Objective—To describe the effects of prednisone and acetylsalicylic acid (ASA) on results of thromboelastography in healthy dogs. Animals—16 male mixed-breed dogs. Procedures—Dogs were randomly assigned to 3 treatment groups (4 dogs/group) that received prednisone (median dose, 2.07 mg/kg), ASA (median dose, 0.51 mg/kg), or both drugs, PO, every 24 hours from days 0 through 6. Another group received no treatment (control dogs; n = 4). Thromboelastography variables (reaction time, clotting time, α-angle, maximum amplitude [MA], global clot strength, coagulation index, and percentage of clot lysis at 60 minutes [CL60]) were evaluated in blood samples collected (prior to drug administration in treated dogs) on days 0 (baseline), 2, 4, and 6. Results—Administration of ASA alone did not alter TEG variables. For treatment effect, mean global clot strength was increased in the prednisone and drug combination groups, compared with values for control dogs; MA was also increased in the prednisone and drug combination groups, compared with that of controls. For treatment-by-time effect, median CL60 was increased in the prednisone group on day 6, compared with baseline value in the same dogs and with median CL60 of the control group on day 6. Median CL60 was also increased in the drug combination group on day 6, compared with the baseline value and with that of the control group on day 6. Conclusions and Clinical Relevance—Prednisone administered at approximately 2 mg/kg/d, PO, for 7 days with or without concurrently administered ASA increased clot strength and decreased clot lysis in healthy dogs.

This study evaluated the analgesia effects of the epidural administration of 0.1 mg/kg bodyweight (BW) of morphine or 5 micrograms/kg BW of buprenorphine in ponies with radiocarpal joint synovitis. Six ponies were submitted to 3 epidural treatments: the control group (C) received 0.15 mL/kg BW of a 0.9% sodium chloride (NaCl) solution; group M was administered 0.1 mg/kg BW of morphine; and group B was administered 5 micrograms/kg BW of buprenorphine, both diluted in 0.9% NaCl to a total volume of 0.15 mL/kg BW administered epidurally at 10 s/mL. The synovitis model was induced by injecting 0.5 ng of lipopolysaccharide (LPS) in the left or right radiocarpal joint. An epidural catheter was later introduced in the lumbosacral space and advanced up to the thoracolumbar level. The treatment started 6 h after synovitis induction. Lameness, maximum angle of carpal flexion, heart rate, systolic arterial pressure, respiratory rate, temperature, and intestinal motility were evaluated before LPS injection (baseline), 6 h after LPS injection (time 0), and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after treatments. Although the model of synovitis produced clear clinical signs of inflammation, the lameness scores in group C were different from the baseline for only up to 12 h. Both morphine and buprenorphine showed a reduction in the degree of lameness starting at 0.5 and 6 h, respectively. Reduced intestinal motility was observed at 0.5 h in group M and at 0.5 to 1 h in group B. Epidural morphine was a more effective analgesic that lasted for more than 12 h and without side effects. It was concluded that morphine would be a valuable analgesic option to alleviate joint pain in the thoracic limbs in ponies.

In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class Ⅰ, lower internalization than that of B. abortus 544; class Ⅱ, similar internalization to that of that of B. abortus 544; class Ⅲ, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class Ⅲ showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

Lack of hygiene and puerperal mastitis are common causes of bacterial diseases in nursing neonates. The aim of this study was to isolate microorganisms from milk samples of healthy female Jindo dogs with suckling puppies and to investigate antimicrobial susceptibility against the isolated bacteria. Milk samples were collected from 120 udders of 12 lactating Jindo dogs that were 2~4 years old without any clinical diseases including mastitis. Bacteria were isolated from 64 milk samples (53.3%), either singly (76.6%) or in combination (23.4%). Staphylococcus (S.) spp. was the most common microorganisms (74.7%) isolated from canine milk, followed by Haemophillus spp. (10.9%), Streptococcus spp. (9.6%), Gardnerella spp. (2.4%) and Moraxella spp. (2.4%). The most frequently isolated organism was S. warneri (31.3%). Antimicrobial susceptibility of these bacteria was tested with 17 antimicrobial agents by Kirbyand Bauer standardized disc diffusion method. Results indicated that bacteria isolated from healthy canine milk were mostly susceptible to amoxicillin-clavulanic acid, cephalothin and ceftiofur, but were resistant to erythromycin, neomycin and tetracycline.