Objective: To determine the effects of raw meat–based diets with and without inulin or yeast cell-wall (YCW) extract on macronutrient digestibility, blood cell counts, serum metabolite concentrations, and fecal fermentative end-product concentrations in healthy adult dogs. Animals: 6 healthy adult spayed female dogs (mean ± SD age, 5.5 ± 0.5 years; mean body weight, 8.5 ± 0.5 kg). Procedures: Dogs were fed each of the following 6 diets for 21 days, the order of which was randomly assigned in a Latin square design: beef control, beef and 1.4% inulin, beef and 1.4% YCW extract, chicken control, chicken and 1.4% inulin, and chicken and 1.4% YCW extract. Each diet trial consisted of a phase for diet adaptation (days 0 to 14) and a phase for measurement of urine and fecal output and content (days 15 to 20). On day 21, food was withheld for blood sample collection. Afterward, the next diet trial began immediately. Results: All dogs maintained desirable fecal quality characteristics and produced low fecal volume. All diets were highly digestible (protein digestibility > 88%; fat digestibility > 97%). Differences in fermentative end-product concentrations among all diets were minor, but a significant increase in fecal short-chain fatty acid concentrations was evident when dogs were fed beef-based diets with inulin and YCW extract. Fecal spermine concentrations were higher with diets containing inulin and YCW extract than with control diets. Blood cell counts and serum metabolite values were within reference limits after each trial. All diets resulted in maintenance of nitrogen balance. Conclusions and Clinical Relevance: Results suggested the raw meat–based diets evaluated were highly digestible in dogs. The increase in fecal short-chain fatty acid concentrations achieved when inulin and YCW extract were included may be beneficial to canine health.

Objective: To determine the oncolytic efficacy of an attenuated form of myxoma virus lacking the serp2 gene in canine tumor cells. Sample: Primary cells were isolated from tumors that were surgically removed from dogs and from connective tissue obtained from the cadaver of a dog. Cells of various established cell lines from tumors and nontumorous tissues were obtained. Procedures: Experiments were performed with cells in monolayer culture. Cell cultures were inoculated with wild-type myxoma viruses or myxoma viruses lacking the serp2 gene, and measures of cytopathic effects, viral growth kinetics, and cell death and apoptosis were determined. Results: Myxoma viruses replicated in cells of many of the primary and established canine tumor cell lines. Canine tumor cells in which expression of activated protein kinase B was upregulated were more permissive to myxoma virus infection than were cells in which expression of activated protein kinase B was not upregulated. Myxoma viruses lacking the serp2 gene caused more cytopathic effects in canine tumor cells because of apoptosis than did wild-type myxoma viruses. Conclusions and Clinical Relevance: Results of the present study indicated myxoma viruses lacking the serp2 gene may be useful for treatment of cancer in dogs. Impact for Human Medicine: Results of the present study may be useful for development of novel oncolytic treatments for tumors in humans.

Objective: To evaluate a human radioimmunoassay (RIA) and equine and high-range porcine (hrp) species-specific ELISAs for the measurement of high serum insulin concentrations in ponies. Samples: Serum samples from 12 healthy nonobese ponies (7 clinically normal and 5 laminitis prone; 13 to 26 years of age; 11 mares and 1 gelding) before and after glucose, insulin, and dexamethasone administration. Procedures: Intra-and interassay repeatability, freeze-thaw stability, dilutional parallelism, and assay agreement were assessed. Results: Assay detection limits were as follows: RIA, < 389 μU/mL; equine ELISA, < 175 μU/mL; and hrp ELISA, 293 to 8,775 μU/mL. Mean ± SD intra- and interassay repeatability were respectively as follows: RIA, 6.5 ± 5.1 % and 74 ± 3.4%; equine ELISA, 10.6 ± 11.0% and 9.0 ± 4.6%; and hrp ELISA, 19.9 ± 172% and 173 ± 16.6%. Freezing and thawing affected measured concentrations. Dilutional parallelism in the RIA was only evident when insulin-depleted equine serum was used as a diluent (percentage recovery, 95.7 ± 274%); in the ELISAs, dilutional parallelism was observed when a zero calibrator was used. Agreement between RIA and equine ELISA results was good for samples containing concentrations < 175 μU of insulin/mL (bias, −18.5 ± 25.5 μU/mL; higher in RIA). At higher concentrations, assay agreement was poor between RIA and equine ELISA results (bias, −185.3 ± 98.7 μU/mL) and between RIA and hrp ELISA results (bias, 25.3 ± 183.0 μU/mL). Conclusions and Clinical Relevance: Agreement among results of the 3 assays was variable, and dilutional parallelism was only evident with the RIA when insulin-depleted equine serum was tested. Caution is recommended when evaluating high insulin concentrations measured with the RIA or ELISAs.

Objective: To identify suitable reference genes for normalization of real-time quantitative PCR (RT-qPCR) assay data for common tumors of dogs. Sample: Malignant lymph node (n = 8), appendicular osteosarcoma (9), and histiocytic sarcoma (12) samples and control samples of various nonneoplastic canine tissues. Procedures: Array-based comparative genomic hybridization (aCGH) data were used to guide selection of 9 candidate reference genes. Expression stability of candidate reference genes and 4 commonly used reference genes was determined for tumor samples with RT-qPCR assays and 3 software programs. Results: LOC611555 was the candidate reference gene with the highest expression stability among the 3 tumor types. Of the commonly used reference genes, expression stability of HPRT was high in histiocytic sarcoma samples, and expression stability of Ubi and RPL32 was high in osteosarcoma samples. Some of the candidate reference genes had higher expression stability than did the commonly used reference genes. Conclusions and Clinical Relevance: Data for constitutively expressed genes with high expression stability are required for normalization of RT-qPCR assay results. Without such data, accurate quantification of gene expression in tumor tissue samples is difficult. Results of the present study indicated LOC611555 may be a useful RT-qPCR assay reference gene for multiple tissue types. Some commonly used reference genes may be suitable for normalization of gene expression data for tumors of dogs, such as lymphomas, osteosarcomas, or histiocytic sarcomas.

Objective: To determine various measurements of medial retropharyngeal lymph nodes (MRPLNs) in healthy cats via ultrasonography and CT. Animals: 45 cats (age range, 2 to 8 years). Procedures: Cats underwent CT of the head and ultrasonography of the cervical region. Various measurements of MRPLNs were obtained, and parenchymal heterogeneity, presence of a hilus, appearance of margins, and attenuation of MRPLNs were determined. Results: Data for 7 cats were excluded because they did not meet inclusion criteria; data for 38 cats were evaluated. Measurements of left and right MRPLNs were not significantly different. Mean length × rostral height × rostral width dimensions of MRPLNs were 20.7 × 12.4 × 3.7 mm and 20.7 × 13.1 × 4.7 mm in ultrasonographic and CT images, respectively. Maximum MRPLN dimensions were approximately 32 × 20 × 7 mm. Mean attenuation of MRPLNs was 40.2 Hounsfield units. Parenchyma of MRPLNs was mildly (via CT) to moderately (via ultrasonography) heterogeneous. A hilus was identified in 95% (via ultrasonography) and 24% or 92% (via CT [depending on criteria used to define a hilus]) of MPRLNs. Lymph node margins were smooth in CT images and mildly irregular in ultrasonographic images. A negative linear correlation was detected between age of cat and MRPLN volume. Conclusions and Clinical Relevance: MRPLNs in cats were easily imaged via ultrasonography and CT. Left and right MRPLNs were symmetric, and MRPLNs were larger in young adult cats versus old cats. Data were intended to serve as references for evaluation of MRPLNs in healthy cats.

Objective: To determine mechanical properties of various prosthetic materials secured to cadaveric canine femurs via various methods and to compare results with those for isolated loops of prosthetic material. Sample: 80 femurs obtained from cadavers of skeletally mature large-breed dogs. Procedures: 10 femoral constructs in each of 8 groups (single circumfabellar loop of polyethylene cord, double loop of polyethylene tape secured via a bone anchor [BAPT], single or double circumfabellar loops of nylon leader material [CNL] or polyethylene tape [CPT], or single or double loops of polyethylene tape secured via a toggle placed through a bone tunnel [BTPT]) and 10 isolated loops of prosthetic material in each corresponding configuration were tested. Stress relaxation, creep, elongation, load at 3 mm of displacement, stiffness, and peak load at failure were determined. Results: 5 single CNL constructs failed before completion of testing. Double CNL and single circumfabellar polyethylene cord constructs had the lowest loads at 3 mm of displacement. Single and double CPT constructs had the highest stiffness. Double BTPT and CPT constructs had the highest peak loads at failure. Double BTPT, double CPT, and BAPT constructs were mechanically superior on the basis of lower creep and stress relaxation and higher stiffness and load at 3 mm of displacement versus other constructs. Stiffness of femoral constructs was 28% to 69% that of corresponding isolated prosthetic loops. Conclusions and Clinical Relevance: Double BTPT, double CPT, and BAPT constructs were mechanically superior to other constucts. Mechanical properties and methods of anchorage and securing of free ends of prostheses contributed to mechanical properties of constructs.

Objective: To determine concentration-dependent effects of tiludronate on cartilage explants incubated with or without recombinant equine interleukin-1β (rEq IL-1). Sample: Articular cartilage explants from the femorotibial joints of 3 young adult horses. Procedures: Cartilage explants were incubated with 1 of 6 concentrations (0, 0.19, 1.9, 19, 190, or 1,900 mg/L) of tiludronate and with or without rEq IL-1 (0.01 ng/mL) for 96 hours. Prostaglandin E2 (PGE2) concentrations in culture medium and explant digests were analyzed via PGE2 enzyme immunoassay. Sulfated glycosaminoglycan (sGAG) concentrations in culture medium were quantified via 1,9-dimethylmethylene blue assay. Chondrocyte apoptosis in paraffin embedded explant sections was measured via terminal deoxynucleotidyl transferase-mediated dUTP nick end–labeling assay. Relative gene expression of matrix metalloproteinases (MMPs), interleukin (IL)-6, and IL-8 was determined via the comparative cycle threshold method. Results: rEq IL-1 increased PGE2 concentration, sGAG release from explants, chondrocyte apoptosis, and MMP gene expression. Lower tiludronate concentrations reduced rEq IL-1–induced sGAG release and chondrocyte apoptosis, whereas the higher tiludronate concentrations increased sGAG release and chondrocyte apoptosis. At the highest tiludronate concentration evaluated, IL-8 gene expression was increased independent of whether rEq IL-1 was present. Conclusions and Clinical Relevance: Tiludronate had biphasic concentration-dependent effects on cartilage explants that were independent of PGE2 secretion or MMP gene expression. Low tiludronate concentrations had some chondroprotective effects, whereas high tiludronate concentrations were detrimental to equine articular cartilage. Administration of tiludronate intra-articularly to horses may be detrimental, dependent on the dose used. In vivo studies are needed before intra-articular tiludronate administration to horses can be recommended.

Objective: To assess joint kinematics in dogs with osteoarthritis of the hip joints during walking up an incline or down a decline and over low obstacles and to compare findings with data for nonlame dogs. Animals: 10 dogs with osteoarthritis of the hip joints (mean ± SD age, 6.95 ± 3.17 years; mean body weight, 34.33 ± 13.58 kg) and 8 nonlame dogs (3.4 ± 2.0 years; 23.6 ± 4.6 kg). Procedures: Reflective markers located on the limbs and high-speed cameras were used to record joint kinematics during walking up an incline or down a decline and over low obstacles. Maximal flexion, extension, and range of motion of the hip joints were calculated. Results: Osteoarthritis of the hip joints reduced extension of both hip joints and flexion of the contralateral hind limb, compared with flexion of the lame hind limb, during walking down a decline. Walking up an incline resulted in decreased extension of the stifle joint in both hind limbs of osteoarthritic dogs; extension was significantly decreased for the lame hind limb. During walking over low obstacles, maximal flexion of the stifle joint was increased significantly for the contralateral hind limb. Maximal flexion was increased in both tarsal joints. Conclusions and Clinical Relevance: Osteoarthritis of the hip joints led to complex changes in the gait of dogs, which involved more joints than the affected hip joint alone. Each exercise had specific effects on joint kinematics that must be considered when planning a rehabilitation program.

Objective: To determine the influence of intensified training and subsequent reduced training on glucose metabolism rate and peripheral insulin sensitivity in horses and identify potential markers indicative of early overtraining. Animals: 12 Standardbred geldings. Procedures: Horses underwent 4 phases of treadmill-based training. In phase 1, horses were habituated to the treadmill. In phase 2, endurance training was alternated with high-intensity exercise training. In phase 3, horses were divided into control and intensified training groups. In the intensified training group, training intensity, duration, and frequency were further increased via a protocol to induce overtraining; in the control group, these factors remained unaltered. In phase 4, training intensity was reduced. Standardized exercise tests were performed after each phase and hyperinsulinemic euglycemic clamp (HEC) tests were performed after phases 2, 3, and 4. Results: 10 of 12 horses completed the study. Dissociation between mean glucose metabolism rate and mean glucose metabolism rate-to-plasma insulin concentration ratio (M:I) was evident in the intensified training group during steady state of HEC testing after phases 3 and 4. After phase 4, mean glucose metabolism rate was significantly decreased (from 31.1 ± 6.8 μmol/kg/min to 18.1 ± 3.4 μmol/kg/min), as was M:I (from 1.05 ± 0.31 to 0.62 ± 0.17) during steady state in the intensified training group, compared with phase 3 values for the same horses. Conclusions and Clinical Relevance: Dissociation between the glucose metabolism rate and M:I in horses that underwent intensified training may reflect non-insulin–dependent increases in glucose metabolism.

Objective: To characterize acute inflammatory and hemostatic surgical stress responses following castration in cats and to evaluate whether the addition of local anesthesia to the anesthetic protocol attenuates these responses. Animals: 39 male cats. Procedures: Cats undergoing castration were randomly assigned to 2 groups: both groups underwent surgery with general anesthesia, and 1 group additionally received a local anesthetic (lidocaine [2.0 mg/kg in total, divided intratesticularly and SC]) prior to incision. Blood samples were collected after anesthetic induction (baseline) and 1, 5, and 24 hours later. Thromboelastography and coagulation variables (activated partial thromboplastin time [aPTT] and prothrombin time [PT]) were analyzed; fibrinolysis was assessed with plasma D-dimer concentrations. The acute-phase response was evaluated via measurement of plasma fibrinogen and serum amyloid A (last time point, 28 hours) concentrations. Hematologic variables were analyzed at baseline and 1, 5, and 24 hours later. Results: Evidence of hemostatic and inflammatory activation after surgery was detected in both groups. Maximum amplitude and G (global clot strength) were significantly increased at 24 hours, and significant, but not clinically relevant, decreases were detected in aPTT at 5 and 24 hours and in PT at 24 hours, compared with baseline values. Serum amyloid A concentrations were significantly higher at 24 and 28 hours than at baseline, and plasma fibrinogen concentration was significantly increased at 24 hours; WBC and RBC counts and Hct were significantly increased at multiple time points. No differences between groups were detected for any variables. Conclusions and Clinical Relevance: Castration appeared to induce hypercoagulability and an acute-phase inflammatory response in cats. Local anesthesia with lidocaine did not attenuate this response.