OBJECTIVE To evaluate the eosinophilic response in intestinal mucosa of horses with intestinal ischemia and reperfusion or with strangulation of the jejunum or colon. SAMPLE Mucosal samples from horses with naturally occurring strangulation (n = 24 horses) or distention (n = 6) of the jejunum or colon (11), with experimentally induced ischemia and reperfusion of the jejunum (6) or colon (15), or that were euthanized for reasons other than gastrointestinal tract disease (13). PROCEDURES Mucosal samples were collected and grouped by type of intestinal injury. Slides were stained with Luna eosinophil stain and histologically examined to determine eosinophil accumulation and distribution. Number of eosinophils per mm2 of mucosa was calculated as a measure of eosinophil accumulation. Additionally, mucosa was categorized into 5 regions; the percentage of eosinophils in each of the 5 regions, relative to the total eosinophil count in all regions, was determined. RESULTS Eosinophil migration toward and onto the luminal surface was evident in tissues after ischemia and reperfusion and after naturally occurring strangulating disease of the jejunum and colon, as indicated by a decrease in the number of eosinophils near the muscularis mucosa and an increase in the number of eosinophils on or near the luminal surface. Ischemia alone did not change eosinophil distribution in the jejunum or colon. CONCLUSIONS AND CLINICAL RELEVANCE Eosinophils responded to mucosal damage evoked by ischemia and reperfusion by migration toward and onto the luminal surface. This migration could represent an important component of the inflammatory response to injury in equine gastrointestinal mucosa.

OBJECTIVE To determine effects of in ovo administration of a probiotic on development of the intestinal microbiota of 2 genetic lineages (modern and heritage) of chickens. SAMPLE 10 newly hatched chicks and 40 fertile eggs to determine intestinal microbiota at hatch, 900 fertile eggs to determine effects of probiotic on hatchability, and 1,560 chicks from treated or control eggs. PROCEDURES A probiotic competitive-exclusion product derived from adult microbiota was administered in ovo to fertile eggs of both genetic lineages. Cecal contents and tissues were collected from embryos, newly hatched chicks, and chicks. A PCR assay was used to detect bacteria present within the cecum of newly hatched chicks. Fluorescence in situ hybridization and vitality staining were used to detect viable bacteria within intestines of embryos. The intestinal microbiota was assessed by use of 16S pyrosequencing. RESULTS Microscopic evaluation of embryonic cecal contents and tissues subjected to differential staining techniques revealed viable bacteria in low numbers. Development of the intestinal microbiota of broiler chicks of both genetic lineages was enhanced by in ovo administration of adult microbiota. Although the treatment increased diversity and affected composition of the microbiota of chicks, most bacterial species present in the probiotic were transient colonizers. However, the treatment decreased the abundance of undesirable bacterial species within heritage lineage chicks. CONCLUSIONS AND CLINICAL RELEVANCE In ovo inoculation of a probiotic competitive-exclusion product derived from adult microbiota may be a viable method of managing development of the microbiota and reducing the prevalence of pathogenic bacteria in chickens.

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 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.

OBJECTIVE To determine values for tear production, horizontal palpebral fissure length (HPFL), eye blink frequency, and intraocular pressure (IOP) in healthy Syrian hamsters (Mesocricetus auratus). ANIMALS 40 healthy adult Syrian hamsters (80 eyes). PROCEDURES Tear production was measured with the phenol red thread test (PRTT), modified Schirmer tear test (mSTT), and endodontic absorbent paper points tear test (EAPPTT). The IOP was measured by use of rebound tonometry. Correlations between test results and body weight were evaluated. RESULTS Mean ± SD values for the IOP, PRTT, EAPPTT, mSTT, HPFL, and blink frequency for all 80 eyes were 4.55 ± 1.33 mm Hg, 5.57 ± 1.51 mm/15 s, 4.52 ± 1.55 mm/min, 2.07 ± 0.97 mm/min, 5.84 ± 0.45 mm, and 1.68 ± 0.43 blinks/min, respectively. For all variables, values did not differ significantly between the right and left eyes or between males and females. There was no correlation between measured variables and body weight.CONCLUSIONS AND CLINICAL RELEVANCE Results for this study provided information on values for the IOP, PRTT, mSTT, EAPPTT, HPFL, and eye blink frequency in healthy Syrian hamsters. It was important to determine reference intervals for this species because they commonly are kept as pets or used as research animals.

OBJECTIVE To evaluate contrast agents for their ability to improve visualization of the colon wall and lumen during CT and ultrasonography. ANIMALS 10 healthy adult Beagles. PROCEDURES Food was withheld from dogs for 36 hours, after which dogs consumed 250 mL of polyethylene glycol solution. Dogs were then anesthetized, a contrast agent (tap water, diluted barium, or air; order randomly assigned) was administered rectally, iodine contrast medium (880 mg of I/kg) was administered IV, and CT and ultrasonography of the colon were performed. After a 1-week washout period, this process was repeated with a different contrast agent until all agents had been evaluated. Two investigators reviewed the CT and ultrasonographic images for colon wall thickness, conspicuity, artifacts, wall layering, and degree of lumen dilation at 4 sites. RESULTS Thickness of the colon wall was greatest in CT and ultrasonographic images with water used as contrast agent, followed by barium and then air. The CT images obtained after water administration had a smooth appearance that outlined the colonic mucosa and had the highest score of the 3 contrast agents for wall conspicuity. Although no substantial artifacts related to any of the contrast agents were identified on CT images, barium- and gas-induced shadowing and reverberation artifacts hindered wall evaluation during ultrasonography. For ultrasonography, the degree of conspicuity was highest with barium in the near-field wall and with water in the far-field wall. In contrast to CT, ultrasonography could be used to distinguish wall layering, and the mucosal and muscular layers were distinct with all contrast agents. CONCLUSIONS AND CLINICAL RELEVANCE Use of water as a contrast agent for both CT and ultrasonography of the colon in dogs compensated for each imaging modality's disadvantages and could be beneficial in the diagnosis of colon disease.

Adipose-derived stem cells (ADSCs) are multipotent, and can be differentiated into many cell types in vitro. In this study, tissues from pigs were chosen to identify and characterize ADSCs. Primary ADSCs were sub-cultured to passage 28. The surface markers of ADSCs: CD29, CD71, CD73, CD90, and CD166 were detected by reverse-transcription polymerase chain reaction assays and the markers CD29, CD44, CD105, and vimentin were detected by immunofluorescence. Growth curves and the capacity of clone-forming were performed to test the proliferation of ADSCs. Karyotype analysis showed that ADSCs cultured in vitro were genetically stable. To assess the differentiation capacity of the ADSCs, cells were induced to differentiate into osteoblasts, adipocytes, epithelial cells, neural cells, and hepatocyte-like cells. The results suggest that ADSCs from pigs showed similar biological characteristics with those separated from other species, and their multi-lineage differentiation shows potential as an application for cellular therapy in an animal model.

OBJECTIVE To characterize a population of Brazilian minipigs with naturally occurring syndactyly by use of plain radiographs and CT images and to evaluate kinetic and temporospatial variables by use of a pressure-sensing walkway. ANIMALS 10 Brazilian minipigs from 6 to 8 months of age (group 1, 5 healthy pigs [body weight, 10.5 to 18.5 kg]; group 2, 5 pigs with syndactyly [body weight, 7.5 to 18.0 kg]). PROCEDURES Forelimbs and hind limbs of all pigs were assessed by use of radiography and CT. Gait was analyzed by use of a pressure-sensing walkway. RESULTS All limbs of all pigs of group 2 had syndactyly. Two forelimbs had complex-1 syndactyly, and 8 forelimbs had complex-2 syndactyly. Four hind limbs had simple syndactyly, 1 hind limb had complex-1 syndactyly, and 5 hind limbs had complex-2 syndactyly. Kinetic and temporospatial values and symmetry indices did not differ between groups. Plantar and palmar surfaces of healthy pigs had 2 areas of maximum pressure, whereas plantar and palmar surfaces of pigs with syndactyly had only 1 area of maximum pressure. CONCLUSIONS AND CLINICAL RELEVANCE In this population of pigs, the most common type of syndactyly was complex-2, and comparison with the healthy group revealed no alteration in kinetic and temporospatial variables. Therefore, results suggested that syndactyly in young minipigs did not cause locomotor disturbances.

OBJECTIVE To measure the minimal joint space width (mJSW) in caudocranial radiographic views of orthopedically normal femorotibial joints of horses, to compare the accuracy of measurements with those of a software program designed for humans, and to identify the ideal caudocranial radiographic projection angle for mJSW measurement. ANIMALS 12 healthy mares (22 femorotibial joints) and 3 equine cadavers (6 stifle joints). PROCEDURES Caudocranial views of femorotibial joints were acquired in the proximodistal plane at 5°, 10°, and 15° (caudo-5°-proximal-craniodistal oblique, 10°, and 15°) and lateromedial plane (caudo-10°-proximo-5°-lateral-craniodistomedial oblique and caudo-10°-proximo-5°-medial-craniodistolateral oblique). The mJSWs of medial and lateral femorotibial joint compartments were measured manually by 2 evaluators and automatically by a digital analysis software program. Interevaluator reproducibility was assessed. Post hoc tests were used to identify the projection angle that provided the largest measurements. Validation of mJSW measurements was performed by evaluation of 6 stifle joints ex vivo. RESULTS Excellent agreement was achieved between the 2 evaluators and between the veterinary radiologist and the analysis software for the medial and lateral compartments of femorotibial joints. Angle of caudocranial view in the proximodistal but not lateromedial plane had a significant effect on the medial compartment mJSW measurements. Mean mJSW for the medial compartment was significantly higher for the caudoproximal-craniodistal oblique projection made at 10° from the horizontal than for other angles. Angle had no significant effect on mean mJSW for the lateral compartment. Agreement between automated measurements of mJSW in the medial compartment and thickness of nonmineralized cartilage in histologic preparations of associated tissues was excellent. CONCLUSIONS AND CLINICAL RELEVANCE Measurements of mJSW in the medial compartment of femorotibial joints, the most common site of osteoarthritis in horses, were reproducible and optimal with a caudoproximal-craniodistal oblique radiographic projection made at 10° from the horizontal.