Objective-To evaluate tendon injuries in horses over a 16-week period by use of ultrasonography and low-field magnetic resonance imaging (MRI). Sample-Tendons of 8 young adult horses. Procedures-The percentage of experimentally induced tendon injury was evaluated in cross section at the maximal area of injury by use of ultrasonography and MRI at 3, 4, 6, 8, and 16 weeks after collagenase injection. The MRI signal intensities and histologic characteristics of each tendon were determined at the same time points. Results-At 4 weeks after collagenase injection, the area of maximal injury assessed on cross section was similar between ultrasonography and MRI. In lesions of > 4 weeks' duration, ultrasonography underestimated the area of maximal cross-sectional injury by approximately 18%, compared with results for MRI. Signal intensity of lesions on T1-weighted images was the most hyperintense of all the sequences, lesions on short tau inversion recovery images were slightly less hyperintense, and T2-weighted images were the most hypointense. Signal intensity of tendon lesions was significantly higher than the signal intensity for the unaltered deep digital flexor tendon. Histologically, there was a decrease in proteoglycan content, an increase in collagen content, and minimal change in fiber alignment during the 16 weeks of the study. Conclusions and Clinical Relevance-Ultrasonography may underestimate the extent of tendon damage in tendons with long-term injury. Low-field MRI provided a more sensitive technique for evaluation of tendon injury and should be considered in horses with tendinitis of > 4 weeks' duration.

Objective-To investigate shedding of chlamydiae from conjunctiva and genital tracts of cats without clinical signs of conjunctivitis or other infectious disease in relation to their titers of serum antibodies against chlamydiae and to serum amyloid A (SAA) and serum alpha1-acid glycoprotein (AGP) concentrations. Animals-62 healthy cats. Procedures-Serum from each cat was analyzed for antibodies against chlamydiae and for SAA and AGP concentrations. Swab samples from the conjunctival sac and genital tract were analyzed with a real-time PCR assay for Chlamydiaceae. Results-4 of 8 of cats with high antibody titers (ie, 1,600) shed chlamydiae, but only from the conjunctiva. Chlamydiae could not be detected in samples from cats with lower antibody titers nor from any genital tract samples. In cats with antibody titers of 1,600, mean +/- SD SAA concentration was significantly higher when chlamydiae were detected in conjunctival swab samples (3.9 +/- 1.0 mg/L) than when no chlamydiae were detected (1.4 +/- 1.0 mg/L). However, SAA concentration was greater than the limit for an acute-phase response in only one of those cats. There was no significant difference in serum AGP concentrations between cats with high titers that were or were not shedding chlamydiae. Nine of 30 (30%) cats (5 with and 4 without detectable serum antibodies against chlamydiae) that had been mated developed reproductive disorders. Conclusions and Clinical Relevance-Clinically normal cats with high chlamydiae-specific antibody titers can shed and thus transmit chlamydiae. Venereal spread from cats without clinical signs of infection is likely not common.

Objective—To evaluate molecular and histologic characteristics of the superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFT), and suspensory ligament (SL) and assess trace-mineral concentrations in serum, liver, and hair of juvenile llamas with metacarpophalangeal and metatarsophalangeal joint hyperextension. Animals—12 juvenile llamas (6 with bilateral hyperextension of metacarpophalangeal joints, metatarsophalangeal joints, or both and 6 clinically normal control llamas). Procedures—Radiography and ultrasonography of metacarpophalangeal and metatarsophalangeal regions were performed. Llamas were euthanized, and SDFT, DDFT, and SL samples were collected for histologic evaluation of collagen and elastin content and orientation, proteoglycan content, and collagen type III immunohistochemistry. Total RNA was isolated from SL tissue, and gene expression of collagen types I and III, lysyl oxidase, and matrix metalloproteinase-13 was evaluated via real-time quantitative reverse transcriptase PCR assay. Liver, serum, and hair samples were evaluated for trace mineral content. Results—Collagen type III gene expression and proteoglycan content were significantly increased in SL samples of affected juvenile llamas, compared with those of control llamas. No difference was detected in collagen and elastin content and orientation or in gene expression of collagen type I, lysyl oxidase, or matrix metalloproteinase-13 between groups. Affected llamas had significantly increased serum molybdenum and decreased liver cobalt concentrations, compared with values for control llamas. Conclusions and Clinical Relevance—Increased collagen type III gene expression and proteoglycan content in SL samples of affected juvenile llamas provided evidence of ongoing SL matrix repair. Trace mineral differences may have been attributable to dietary imbalances in affected llamas.

Objective—To evaluate effects of cyclosporine, dexamethasone, and the immunosuppressive agent human CTLA4-Ig on cytokine production by feline lymphocytes in vitro and to assess patterns of cytokine production for 5 immunosuppressed renal transplant recipient cats. Animals—21 clinically normal cats and 5 immunosupressed renal transplant recipient cats. Procedures--Peripheral blood mononuclear cells were isolated from clinically normal cats and stimulated with concanavalin A (Con A; 10 μg/mL) alone or Con A with cyclosporine (0.05 μg/mL), dexamethasone (1 × 10−7M), a combination of cyclosporine-dexamethasone, or human CTLA4-Ig (10 g/mL). Cells from transplant recipients were stimulated with Con A alone. An ELISA was performed to measure production of interferon (IFN)-γ, granulocyte macrophage–colony stimulating factor (GM-CSF), interleukin (IL)-2, IL-4, and IL-10. Proliferation of CD4+ and CD8+T cells from immunosuppressed cats were also evaluated. Pairwise comparisons were performed via a Wilcoxon signed rank test or Wilcoxon rank sum test. Results—Cyclosporine, dexamethasone, cyclosporine-dexamethasone combination, and CTLA4-Ig caused a significant decrease in IL-2, IFN-γ, and GM-CSF production. Cyclosporine and cyclosporine-dexamethasone, but not human CTLA4-Ig, caused a significant decrease in IL-10 production. High basal concentrations of IL-2 and IL-10 were identified in transplant recipients, and IL-10 was significantly increased in stimulated cultures. In immunosuppressed cats, there was a decrease in frequency of responders and proliferative capacity of CD4+ and CD8+T cells. Conclusions and Clinical Relevance—CTLA4-Ig successfully inhibited proinflammatory cytokines while sparing cytokines critical for allograft tolerance. These data may be useful for developing better strategies to prevent rejection while sparing other immune functions

Objective—To evaluate whether an equine-derived canine H3N8 influenza A virus was capable of infecting and transmitting disease to ponies. Animals—20 influenza virus-seronegative 12- to 24-month-old ponies. Procedures—5 ponies were inoculated via aerosol exposure with 10(7) TCID50 of A/Canine/Wyoming/86033/07 virus (Ca/WY)/pony. A second group of 5 ponies (positive control group) was inoculated via aerosol exposure with a contemporary A/Eq/Colorado/10/07 virus (Eq/CO), and 4 sham-inoculated ponies served as a negative control group. To evaluate the potential for virus transmission, ponies (3/inoculation group) were introduced 2 days after aerosol exposure and housed with Ca/WY- and Eq/CO-inoculated ponies to serve as sentinel animals. Clinical signs, nasal virus shedding, and serologic responses to inoculation were monitored in all ponies for up to 21 days after viral inoculation. Growth and infection characteristics of viruses were examined by use of Madin-Darby canine kidney cells and primary equine and canine respiratory epithelial cells. Results—Ponies inoculated with Ca/WY had mild changes in clinical appearance, compared with results for Eq/CO-inoculated ponies. Additionally, Ca/WY inoculation induced significantly lower numbers for copies of the matrix gene in nasal secretions and lower systemic antibody responses in ponies than did Eq/CO inoculation. The Ca/WY isolate was not transmitted to sentinel ponies. Conclusions and Clinical Relevance—Inoculation of ponies with the canine H3N8 isolate resulted in mild clinical disease, minimal nasal virus shedding, and weak systemic antibody responses, compared with responses after inoculation with the equine H3N8 influenza isolate. These results suggested that Ca/WY has not maintained infectivity for ponies.

Objective—To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. Animals—5 healthy adult horses. Procedures—Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20% of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. Results—COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were β-actin and hypoxanthine phosphoribosyltransferase, whereas glyceraldehyde 3-phosphate dehydrogenase and β-2 microglobulin were the least stably expressed. Conclusions and Clinical Relevance—Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.

Objective—To compare recovery of epithelial lining fluid (ELF) in bronchoalveolar lavage fluid (BALF) by use of weight-adjusted or fixed-amount volumes of lavage fluid in dogs. Animals—13 healthy Beagles. Procedures—Dogs were allocated to 2 groups. In 1 group, the right caudal lung lobe was lavaged on the basis of each dog's weight (2 mL/kg, divided into 2 aliquots) and the left caudal lung lobe was lavaged with a fixed amount of fluid (50 mL/dog, divided into 2 aliquots). In the second group, the right and left caudal lung lobes were lavaged by use of the fixed-amount and weight-adjusted techniques, respectively. The BALF was collected by use of bronchoscopy. A recovery percentage ≥ 40% was required. The proportion of ELF was calculated by use of the following equation: (concentration of urea in BALF/concentration of urea in serum) × 100. Results—Mean ± SD proportion of ELF in BALF was 2.28 ± 0.39% for the weight-adjusted technique and 2.89 ± 0.89% for the fixed-amount technique. The SDs between these 2 techniques differed significantly (calculated by comparing 2 covariance structures [unstructured and compound symmetry] in a repeated-measures mixed ANOVA). Conclusions and Clinical Relevance—The findings strongly suggested that use of a weight-adjusted bronchoalveolar lavage technique provided a more uniform ELF recovery, compared with that for a fixed-amount bronchoalveolar lavage technique, when urea was used as a marker of dilution. A constant ELF fraction can facilitate more accurate comparisons of cellular and noncellular constituents in BALF among patients of various sizes.

Objective—To assess validity and reliability for a visual analogue scale (VAS) used by owners to measure chronic pain in their osteoarthritic dogs. Sample—68, 61, and 34 owners who completed a questionnaire. Procedures—Owners answered questionnaires at 5 time points. Criterion validity of the VAS was evaluated for all dogs in the intended-to-treat population by correlating scores for the VAS with scores for the validated Helsinki Chronic Pain Index (HCPI) and a relative quality-of-life scale. Intraclass correlation was used to assess repeatability of the pain VAS at 2 baseline evaluations. To determine sensitivity to change and face validity of the VAS, 2 blinded, randomized control groups (17 dogs receiving carprofen and 17 receiving a placebo) were analyzed over time. Results—Significant correlations existed between the VAS score and the quality-of-life scale and HCPI scores. Intraclass coefficient (r = 0.72; 95% confidence interval, 0.57 to 0.82) for the VAS indicated good repeatability. In the carprofen and placebo groups, there was poor correlation between the 2 pain evaluation methods (VAS and HCPI items) at the baseline evaluation, but the correlation improved in the carprofen group over time. No correlation was detected for the placebo group over time. Conclusions and Clinical Relevance—Although valid and reliable, the pain VAS was a poor tool for untrained owners because of poor face validity (ie, owners could not recognize their dogs' behavior as signs of pain). Only after owners had seen pain diminish and then return (after starting and discontinuing NSAID use) did the VAS have face validity.

Objective—To compare results for 3 commercially available microbiological media plates with those for standard bacteriologic testing of bovine milk. Sample—Milk samples from postpartum cows and cows with a high somatic cell count (SCC) or clinical mastitis (CM). Procedures—Sample-ready Staphylococcus culture medium (SRSC) plates were used to detect Staphylococcus aureus in milk samples obtained from postpartum cows and cows with a high SCC or CM. Rapid coliform count (RCC) plates were used to detect coliforms in milk samples obtained from cows with CM. Aerobic count (AC) plates were used to detect streptococci in CM samples. Fresh mastitic milk samples were frozen and then thawed to evaluate the effects of freezing for the SRSC and RCC plates. The effects of dilution (1:10) of samples were determined. Agreement of results between the commercially available plates and standard bacteriologic testing was evaluated. Results—The ability of SRSC plates to detect S aureus in milk samples was highest with diluted samples from postpartum cows and cows with a high SCC or CM. Sensitivity of the RCC plate for detection of coliforms was highest with diluted mastitic milk samples. The AC plates had a poor positive predictive value for detection of streptococci in mastitic milk samples. Freezing increased S aureus detection. Conclusions and Clinical Relevance—Overall, the SRSC and RCC plates were accurate, were easy to use, and yielded results comparable to those of standard bacteriologic testing for the detection of S aureus and coliforms in bovine milk.

Objective—To investigate the effect of opsonization of Rhodococcus equi with R equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Sample—Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Procedures—Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophage tumor necrosis factor (TNF)-α production was measured via an ELISA. Extracellular and intracellular bacterial viability was measured with a novel R equi-luciferase construct that used a luminometer. Results—Opsonized bacteria increased oxidative burst activity in adult horse phagocytes, and neutrophil activity was dependent on the concentration of specific antibody. Secretion of TNF-α was higher in macrophages infected with opsonized bacteria. Opsonization had no significant effect on bacterial viability in macrophages; however, extracellular bacterial viability was decreased in broth containing plasma with R equi-specific antibodies, compared with viability in broth alone. Conclusions and Clinical Relevance—The use of plasma enriched with specific antibodies for the opsonization of R equi increased the activation of phagocytes and decreased bacterial viability in the extracellular space. Although opsonized R equi increased TNF-α secretion and oxidative burst in macrophages, additional factors may be necessary for effective intracellular bacterial killing. These data have suggested a possible role of plasma antibody in protection of foals from R equi pneumonia.