OBJECTIVE To compare the anticollagenase efficacy of fresh feline, canine, and equine serum and plasma on in vitro corneal degradation. SAMPLE Grossly normal corneas from recently euthanized dogs, cats, and horses and fresh serum and plasma from healthy dogs, cats, and horses. PROCEDURES Serum and plasma were pooled by species and used for in vitro experiments. Corneas were collected and stored at −80°C. Sections of cornea were dried, weighed, and incubated in saline (0.9% NaCl) solution with clostridial collagenase and homologous fresh serum or plasma. Corneal degradation was assessed as the percentage of corneal weight loss and hydroxyproline concentration, compared with results for positive and negative control samples. RESULTS Homologous fresh serum and plasma significantly reduced the percentage of corneal weight loss, compared with results for positive control samples. No significant difference was found in percentage of corneal weight loss between incubation with serum or plasma for feline, canine, and equine corneas. Canine serum and plasma significantly reduced hydroxyproline concentrations, whereas inclusion of feline and equine serum or plasma did not, compared with results for positive control samples. Hydroxyproline concentrations were moderately correlated with percentage of corneal weight loss for feline samples and weakly correlated for equine samples, but they were not correlated for canine samples. CONCLUSIONS AND CLINICAL RELEVANCE In this study, the anticollagenase efficacy of fresh feline, canine, and equine serum was not different from that of plasma. Plasma should be an acceptable substitute for serum in the topical treatment of keratomalacia.

OBJECTIVE To assess the effect of anesthetic induction with a benzodiazepine plus ketamine or propofol on hypothermia in dogs undergoing ovariohysterectomy without heat support. ANIMALS 23 adult sexually intact female dogs undergoing ovariohysterectomy. PROCEDURES Baseline rectal temperature, heart rate, and respiratory rate were recorded prior to premedication with buprenorphine (0.02 mg/kg, IM) and acepromazine (0.05 mg/kg, IM). Anesthesia was induced with midazolam or diazepam (0.25 mg/kg, IV) plus ketamine (5 mg/kg, IV; n = 11) or propofol (4 mg/kg, IV; 12) and maintained with isoflurane in oxygen. Rectal temperature was measured at hospital intake, prior to premedication, immediately after anesthetic induction, and every 5 minutes after anesthetic induction. Esophageal temperature was measured every 5 minutes during anesthesia, beginning 30 minutes after anesthetic induction. After anesthesia, dogs were covered with a warm-air blanket and rectal temperature was measured every 10 minutes until normothermia (37°C) was achieved. RESULTS Dogs in both treatment groups had lower rectal temperatures within 5 minutes after anesthetic induction and throughout anesthesia. Compared with dogs that received a benzodiazepine plus ketamine, dogs that received a benzodiazepine plus propofol had significantly lower rectal temperatures and the interval from discontinuation of anesthesia to achievement of normothermia was significantly longer. CONCLUSIONS AND CLINICAL RELEVANCE Dogs in which anesthesia was induced with a benzodiazepine plus propofol or ketamine became hypothermic; the extent of hypothermia was more profound for the propofol combination. Dogs should be provided with adequate heat support after induction of anesthesia, particularly when a propofol-benzodiazepine combination is administered.

OBJECTIVE To evaluate the biochemical and biomechanical properties of native and decellularized superficial digital flexor tendons (SDFTs) and deep digital flexor tendons (DDFTs) harvested from the pelvic limbs of orthopedically normal dogs. SAMPLE 22 commercially supplied tendon specimens (10 SDFT and 12 DDFT) harvested from the pelvic limbs of 13 canine cadavers. PROCEDURES DNA, glycosaminoglycan, collagen, and protein content were measured to biochemically compare native and decellularized SDFT and DDFT specimens. Mechanical testing was performed on 4 groups consisting of native tendons (5 SDFTs and 6 DDFTs) and decellularized tendons (5 SDFTs and 6 DDFTs). All tendons were preconditioned, and tension was applied to failure at 0.5 mm/s. Failure mode was video recorded for each tendon. Load-deformation and stress-strain curves were generated; calculations were performed to determine the Young modulus and stiffness. Biochemical and biomechanical data were statistically compared by use of the Wilcoxon rank sum test. RESULTS Decellularized SDFT and DDFT specimens had significantly less DNA content than did native tendons. No significant differences were identified between native and decellularized specimens with respect to glycosaminoglycan, collagen, or protein content. Biomechanical comparison yielded no significant intra- or intergroup differences. All DDFT constructs failed at the tendon-clamp interface, whereas nearly half (4/10) of the SDFT constructs failed at midsubstance. CONCLUSIONS AND CLINICAL RELEVANCE Decellularized commercial canine SDFT and DDFT specimens had similar biomechanical properties, compared with each other and with native tendons. The decellularization process significantly decreased DNA content while minimizing loss of extracellular matrix components. Decellularized canine flexor tendons may provide suitable, biocompatible graft scaffolds for bioengineering applications such as tendon or ligament repair.

The aim of the present work was to develop reagents to set up a chicken interferon-γ (ChIFN-γ) assay. Four monoclonal antibodies (mAbs) specific for ChIFN-γ were generated to establish sandwich ELISA based on 2 different mAbs. To improve the detection sensitivity of ChIFN-γ, a double-monoclonal antibody sandwich ELISA was developed using mAb 3E5 as capture antibody and biotinylated mAb 3E3 as a detection reagent. The results revealed that this ELISA has high sensitivity, allowing for the detection of 125 to 500 pg/mL of recombinant ChIFN-γ, and also has an excellent capacity for detecting native ChIFN-γ. This ELISA was then used to detect ChIFN-γ level in chickens immunized with a Newcastle disease virus (NDV) vaccine, the immunized chicken splenocytes were stimulated by NDV F protein as recall antigen. From our results, it appears that the sensitivity range of this sandwich ELISA test is adequate to measure the ex vivo release of ChIFN-γ.

OBJECTIVE To examine ultrasonographic predictors of ovarian development in European eels (Anguilla anguilla) undergoing hormonal treatment for assisted reproduction. ANIMALS 83 female European eels. PROCEDURES Eels received weekly IM injections of salmon pituitary extract (first injection = week 1). Ultrasonography of the ovaries was performed twice during hormonal treatment (weeks 7 and 11). Eels were identified on the basis of body weight as having an adequate response by weeks 14 to 20 or an inadequate response after injections for 21 weeks. Eels were euthanized at the end of the experiment and classified by use of ovarian histologic examination. Ovarian cross-sectional area and size of eel (ie, length3) were used to classify eels (fast responder, slow responder, or nonresponder) and to calculate an ultrasonographic-derived gonadosomatic index. Gray-level co-occurrence matrices were calculated from ovarian images, and 22 texture features were calculated from these matrices. RESULTS The ultrasonographic-derived gonadosomatic index differed significantly between fast responders and slow responders or nonresponders at both weeks 7 and 11. Principal component analysis revealed a pattern of separation between the groups, and partial least squares discriminant analysis revealed signals in the ovarian texture that discriminated females that responded to treatment from those that did not. CONCLUSIONS AND CLINICAL RELEVANCE Ovarian texture information in addition to morphometric variables can enhance ultrasonographic applications for assisted reproduction of eels and potentially other fish species. This was a novel, nonlethal method for classifying reproductive response of eels and the first objective texture analysis performed on ultrasonographic images of the gonads of fish.

OBJECTIVE To describe the perfusion and diffusion characteristics of the liver in healthy dogs as determined by morphological, perfusion-weighted, and diffusion-weighted MRI. ANIMALS 11 healthy adult Beagles. PROCEDURES Each dog was anesthetized and underwent morphological, perfusion-weighted, and diffusion-weighted MRI of the cranial aspect of the abdomen. On the MRI images, a region of interest (ROI) was established for each of 6 structures (aorta, caudal vena cava, portal vein, hepatic parenchyma, splenic parenchyma, and skeletal [epaxial] muscle). The signal intensity was determined, and a time-intensity curve was generated for each ROI. The apparent diffusion coefficient (ADC) was calculated for the hepatic and splenic parenchyma in diffusion-weighted MRI images, and the normalized ADC for the liver was calculated as the ratio of the ADC for the hepatic parenchyma to the ADC for the splenic parenchyma. Dogs also underwent abdominal ultrasonography, and ultrasound-guided fine-needle aspirate samples and biopsy specimens were obtained from the liver for cytologic and histologic examination. RESULTS Cytologic and histologic results suggested that the liver was clinically normal in all dogs. Perfusion-weighted MRI parameters varied among the 6 ROIs. The mean ± SD ADC of the hepatic parenchyma was 0.84 × 10(−3) mm2/s ± 0.17 × 10(−3) mm2/s, and the mean normalized ADC for the liver was 1.8 ± 0.4. CONCLUSIONS AND CLINICAL RELEVANCE Results provided preliminary baseline information about the diffusion and perfusion characteristics of the liver in healthy dogs. Additional studies on dogs of various breeds with and without hepatopathies are necessary to validate and refine these findings.

OBJECTIVE To determine a treatment protocol for SC administration of dalteparin to cats on the basis of currently available detailed pharmacokinetic data and to assess the effect of SC administration of dalteparin to cats on coagulation variables such as activated partial thromboplastin time (aPTT), thrombin time, and results for thromboelastometry, compared with effects on anti–activated coagulation factor X (anti-Xa) activity. ANIMALS 6 healthy domestic shorthair cats. PROCEDURES Cats received 14 injections of dalteparin (75 anti-Xa U/kg, SC) at 6-hour intervals. Blood samples were collected before and 2 hours after the first and second injections on days 1, 2, and 4. Anti-Xa activity was measured by use of a chromogenic substrate assay, aPTT and thrombin time were measured by use of an automated coagulometer, and viscoelastic measurements were obtained with thromboelastrometry. RESULTS 2 hours after the second injection, the target peak anti-Xa activity range of 0.5 to 1.0 U/mL was achieved in all cats, whereas median trough values remained below this range. Peak anti-Xa activity had only minimal effects on coagulation variables; the maximum median ratio for aPTT (in relationship to the value before the first dalteparin injection) was 1.23. CONCLUSIONS AND CLINICAL RELEVANCE Results of this study indicated that this treatment protocol resulted in reproducible anti-Xa activity in cats that was mostly within the targeted peak range of anti-Xa activity recommended for humans. Treatment in accordance with this protocol may not require routine coagulation monitoring of cats, but this must be confirmed in feline patients.

OBJECTIVE To compare the orthogonal diameter (visual metric) method against a manual perimeter tracing (planimetry) method to measure volume of brain tumors in dogs by use of MRI scans. SAMPLE 22 sets of MRI brain scans pertaining to 22 client-owned dogs with histologically confirmed glioma. PROCEDURES MRI scans were reviewed by 2 operators, and scans revealing tumors with a degree of gadolinium enhancement that allowed discrimination between tumor tissue and healthy parenchyma were used. Each operator calculated tumor volume for each set of scans twice by use of visual metric and planimetry methods. Inter- and intraoperator variability were assessed by calculation of an agreement index (AI). RESULTS Mean ± SD intraoperator AIs were 0.79 ± 0.24 for the visual metric method and 0.89 ± 0.17 for the planimetry method. Intraoperator variability for both operators was significantly less when the planimetry method was used than when the visual metric method was used. No significant differences were identified in mean interoperator AI between visual metric (0.68 ± 0.28) and planimetry (0.67 ± 0.31) methods. CONCLUSIONS AND CLINICAL RELEVANCE The lower intraoperator variability achieved with the planimetry versus visual metric method should result in more precise volume assessments when the same operator performs multiple volume measurements of brain tumors in dogs. Equivocal results for interoperator variability may have been due to method variance or inadequate preliminary training. Additional studies are needed to evaluate the suitability of planimetry for assessing response to treatment.

OBJECTIVE To identify cardiac tissue genes and gene pathways differentially expressed between dogs with and without dilated cardiomyopathy (DCM). ANIMALS 8 dogs with and 5 dogs without DCM. PROCEDURES Following euthanasia, samples of left ventricular myocardium were collected from each dog. Total RNA was extracted from tissue samples, and RNA sequencing was performed on each sample. Samples from dogs with and without DCM were grouped to identify genes that were differentially regulated between the 2 populations. Overrepresentation analysis was performed on upregulated and downregulated gene sets to identify altered molecular pathways in dogs with DCM. RESULTS Genes involved in cellular energy metabolism, especially metabolism of carbohydrates and fats, were significantly downregulated in dogs with DCM. Expression of cardiac structural proteins was also altered in affected dogs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that RNA sequencing may provide important insights into the pathogenesis of DCM in dogs and highlight pathways that should be explored to identify causative mutations and develop novel therapeutic interventions.

The objective of this study was to compare the efficacy of 2 different commercial Mycoplasma hyopneumoniae vaccines and porcine reproductive and respiratory syndrome virus (PRRSV) vaccines in regard to growth performance, microbiological and immunological analyses, and pathological observation from wean to finish (175 d of age). Pigs were administered M. hyopneumoniae and PRRSV vaccines at 7 and 21 d of age, respectively, or both at 21 d old and then challenged with both M. hyopneumoniae and PRRSV at 49 d old. Significant (P < 0.05) differences were observed between the 2 vaccinated challenged groups in average daily weight gain, nasal shedding of M. hyopneumoniae, M. hyopneumoniae-specific interferon-γ secreting cells, and macroscopic and microscopic lung lesions. Induction of interleukin-10 following PRRSV vaccination does not interfere with the immune responses induced by M. hyopneumoniae vaccine. The present study demonstrated that the single-dose vaccination regimen for M. hyopneumoniae and PRRSV vaccine is efficacious for controlling coinfection with M. hyopneumoniae and PRRSV based on clinical, microbiological, immunological, and pathological evaluation.