Tissues unsuitable for standard immunohistochemical and histopathological examinations for chronic wasting disease (CWD) in cervids and for scrapie in sheep are frequently submitted for testing. This study investigated the effects of experimental autolysis on the detection of abnormal prion protein (PrPsc) in lymphoid and central nervous system (CNS) tissues from elk and sheep. The PrPsc was detected using a Western blotting (WB) test following PrPsc enrichment using sodium phosphotungstic acid (PTA) precipitation (PTA-WB). A commercial enzyme-linked immunosorbent assay (ELISA) was used as a reference test for quantitative measurement. This study showed that the amount of PrPsc in lymphoid and CNS tssues from elk and sheep decreased gradually as a result of autolysis, but PrPsc was still detectable after 5 and 15 d incubation at 37°C by PTA-WB for all lymphoid and CNS samples. The results of the ELISA supported those of PTA-WB, particularly for CNS tissues. In conclusion, autolysis at 37°C for 15 d would not significantly affect the detection of PrPsc in lymphoid and CNS tissues by WB and ELISA and, particularly, PTA-WB is a valuable and alternative confirmatory test to detect PrPsc in autolyzed lymphoid and CNS samples.

Objective—To investigate the density of the primary epidermal lamellae (PEL) around the solar circumference of the forefeet of near-term fetal feral and nonferal (ie, domesticated) horses. Sample—Left forefeet from near-term Australian feral (n = 14) and domesticated (4) horse fetuses. Procedures—Near-term feral horse fetuses were obtained from culled mares within 10 minutes of death; fetuses that had died in utero 2 weeks prior to anticipated birth date and were delivered from live Thoroughbred mares were also obtained. Following disarticulation at the carpus, the left forefoot of each fetus was frozen during dissection and data collection. In a standard section of each hoof, the stratum internum PEL density was calculated at the midline center (12 o'clock) and the medial and lateral break-over points (11 and 1 o'clock), toe quarters (10 and 2 o'clock), and quarters (4 and 6 o'clock). Values for matching lateral and medial zones were averaged and expressed as 1 density. Density differences at the 4 locations between the feral and domesticated horse feet were assessed by use of imaging software analysis. Results—In fetal domesticated horse feet, PEL density did not differ among the 4 locations. In fetal feral horse feet, PEL density differed significantly among locations, with a pattern of gradual reduction from the dorsal to the palmar aspect of the foot. The PEL density distribution differed significantly between fetal domesticated and feral horse feet. Conclusions and Clinical Relevance—Results indicated that PEL density distribution differs between fetal feral and domesticated horse feet, suggestive of an adaptation of feral horses to environment challenges.

Objective—To evaluate the impact of oxytetracycline exposure on horizontal transfer of an antimicrobial resistance plasmid. Sample—Populations of Salmonella enterica subsp enterica serovar Typhimurium and Escherichia coli. Procedures—Mixed populations of plasmid donor (Salmonella Typhimurium) and recipient (E coli) bacteria were assigned to 1 of 2 simulated oxytetracycline dosing regimens (high peak concentration-short elimination half-life [HC-SHL] or low peak concentration—long elimination half-life [LC-LHL]) or served as untreated control replicates. Donor, recipient, and transconjugant (E coli that acquired the plasmid) bacteria populations were quantified at 12, 24, and 36 hours after oxytetracycline administration by use of culture on selective bacterial growth media. Results—The ratio of transconjugant to donor bacteria was significantly reduced in the oxytetracycline-exposed replicates, compared with the ratio for the control replicates, at 12 hours. At 24 and 36 hours, results for the HC-SHL regimens were not significantly different from results for the respective control replicates, and results for the LC-LHL regimens also were not significantly different from results for the respective control replicates. The oxytetracycline concentration at these time points (12 hours in the HC-SHL regimen and all 3 time points in the LC-LHL regimen) were in excess of the minimum inhibitory concentration of the recipient bacteria. Conclusions and Clinical Relevance—Transfer of antimicrobial resistance plasmids may be suppressed in vitro by oxytetracycline exposure at concentrations greater than the minimum inhibitory concentration of the recipient bacteria.

Objective: To determine the effect of Hct on blood glucose readings of dogs obtained by use of 2 point-of-care (POC) blood glucometers and a laboratory analyzer. Animals: 184 dogs, including 139 Greyhounds. Procedures: Venous blood samples collected from 184 dogs with a range of Hcts (measured in EDTA-anticoagulated blood) were immediately analyzed with a handheld glucometer specifically developed for veterinary use and a glucometer developed for use in humans. The remainder of each blood sample was placed in fluoride oxalate tubes, and plasma glucose concentration was measured with a laboratory analyzer. Agreement between results for the POC glucometers and laboratory analyzer and effect of Hct on glucometer accuracy was assessed via regression analysis. Results: Significant differences were detected between results of the glucometers and the reference laboratory analyzer. The Hct affected the correlation between results for the glucometers and the laboratory analyzer. Deviations of the glucometers from the reference interval varied with Hct. The glucometer for veterinary use more closely correlated with the glucose concentration when Hct was within or above its reference interval. The glucometer for use in humans more closely approximated laboratory reference glucose concentrations in anemic dogs. Conclusions and Clinical Relevance: Hct had a relevant impact on the correlation between whole blood and plasma glucose concentrations in dogs. Significant variations between results obtained with the 2 glucometers could be critical when interpreting blood glucose measurements or selecting a POC glucometer for an intensive care setting and precise glycemic control in critically ill dogs.

Objective: To compare echocardiographic variables of dogs with postmortem anatomic measurements and histologic characteristics of the mitral valve (MV). Animals: 21 cardiologically normal dogs. Procedures: The MV was measured echocardiographically by use of the right parasternal 5-chamber long-axis view. Dogs were euthanized, and anatomic measurements of the MV annulus (MVa) were performed at the level of the left circumflex coronary artery. Mitral valve leaflets (MVLs) and chordae tendineae were measured. Structure of the MVLs was histologically evaluated in 3 segments (proximal, middle, and distal). Results: Echocardiographic measurements of MVL length did not differ significantly from anatomic measurements. A positive correlation was detected between body weight and MVa area. There was a negative correlation between MVa area and the percentage by which the MVL area exceeded the MVa area. Anterior MVLs had a significantly higher number of chordae tendineae than did posterior MVLs. Histologically, layering of MVLs was less preserved in the distal segment, whereas the muscular component and adipose tissue were significantly more diffuse in the proximal and middle segments. Conclusions and Clinical Relevance: The MV in cardiologically normal dogs had wide anatomic variability. Anatomic measurements of MVL length were correlated with echocardiographic measurements.

Objective—To evaluate effects of transplantation of bone marrow stromal cells (BMSCs) into the CSF for the treatment of chronic spinal cord injury in dogs that had not responded by 1 month after decompressive surgery. Animals—23 dogs. Procedures—Dogs with paraplegia and loss of nociception in the pelvic limbs for at least 1 month after decompressive surgery were assigned to transplantation or control groups. Dogs in the transplantation group received BMSCs injected into the CSF 1 to 3 months after decompressive surgery. Dogs in the control group did not receive additional treatments. Improvements in gait, proprioceptive positioning, and nociception were evaluated by use of the Texas Spinal Cord Injury Scale for ≥ 6 months after BMSC transplantation. Results—6 of 10 dogs in the transplantation group regained the ability to walk, whereas only 2 of 13 dogs in the control group regained the ability to walk. Scores for the Texas Spinal Cord Injury Scale in the transplantation group were significantly higher than scores in the control group at the endpoint of the study (6 months after BMSC transplantation or after decompressive surgery for the transplantation and control groups, respectively). Only 1 dog (transplantation group) recovered nociception. All dogs from both groups had fecal and urinary incontinence. No complications were observed in relation to BMSC transplantation. Conclusions and Clinical Relevance—Injection of BMSCs into the CSF caused no complications and could have beneficial effects on pelvic limb locomotion in dogs with chronic spinal cord injuries.

Objective: To assess the effect of computed tomography (CT) scan protocols (radiation amounts) and fabrication methods on biomodel accuracy and variability. Sample: Cadaveric femur of a Basset Hound. Procedures: Retroreconstructions (n = 158) were performed of 16 original scans and were visually inspected to select 17 scans to be used for biomodel fabrication. Biomodels of the 17 scans were made in triplicate by use of 3 freeform fabrication processes (stereolithography, fused deposition modeling, and 3-D printing) for 153 models. The biomodels and original bone were measured by use of a coordinate measurement machine. Results: Differences among fabrication methods accounted for 2% to 29% of the total observed variation in inaccuracy and differences among method-specific radiation configurations accounted for 4% to 44%. Biomodels underestimated bone length and width and femoral head diameter and overestimated cortical thickness. There was no evidence of a linear association between thresholding adjustments and biomodel accuracy. Higher measured radiation dose led to a decrease in absolute relative error for biomodel diameter and for 4 of 8 cortical thickness measurements. Conclusions and Clinical Relevance: The outside dimensions of biomodels have a clinically acceptable accuracy. The cortical thickness of biomodels may overestimate cortical thickness. Variability among biomodels was caused by model fabrication reproducibility and, to a lesser extent, by the radiation settings of the CT scan and differences among fabrication methods.

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 imaging plane, flexion and extension, patient weight, and observer on computed tomographic (CT) image measurements of the area of the lumbosacral (L7-S1) intervertebral foramen (LSIF) in dogs. Sample—12 dog cadavers (2 were excluded because of foraminal stenosis). Procedures—In each cadaver, sagittal, sagittal oblique, transverse oblique, and double oblique CT images were obtained at 3 zones (entrance, middle, and exit zones) of the region of the lateral lumbar spinal canal that comprises the LSIF while the lumbosacral junction (LSJ) was positioned in flexion or extension. Barium-impregnated polymethylmethacrylate was used to fill the intervertebral foramina to aid boundary detection. Measurements of interest were obtained. Results—Among the dog cadavers, there was large variability in LSIF cross-sectional areas (range, 0.12 to 0.44 cm2; SD, 0.1 cm2) and in foraminal angles required to obtain a double oblique plane in LSJ extension (SD, 8° to 9°). For LSIF area measurements in standard sagittal CT images, interobserver variability was 23% to 44% and intraobserver variability was 4% to 5%. Sagittal oblique images obtained during LSJ extension yielded smaller mean LSIF areas (0.30 cm2), compared with findings in sagittal images (0.37 to 0.52 cm2). The exit and middle zone areas were smaller than the entrance zone area in sagittal images obtained during LSJ extension. Conclusions and Clinical Relevance—Repeated measurements of the LSIF area in images obtained during LSJ extension may be unreliable as a result of interobserver variability and the effects of dog positioning and CT slice orientation.

Objective—To compare effects of 4 types of stimulation devices attached to the hind feet on hoof flight, joint angles, and net joint powers of trotting horses. Animals—8 clinically normal horses. Procedures—Horses were evaluated under 5 conditions in random order: no stimulators, loose straps (10 g), lightweight tactile stimulators (55 g), limb weights (700 g), and limb weights with tactile stimulators (700 g). Reflective markers on the hind limbs were tracked during the swing phase of 6 trotting trials performed at consistent speed to determine peak hoof heights and flexion angles of the hip, stifle, tarsal, and metatarsophalangeal joints. Inverse dynamic analysis was used to calculate net joint energies. Comparisons among stimulators were made. Results—Peak hoof height was lowest for no stimulators (mean ± SD, 5.42 ± 1.38 cm) and loose straps (6.72 ± 2.19 cm), intermediate for tactile stimulators (14.13 ± 7.33 cm) and limb weights (16.86 ± 15.93 cm), and highest for limb weights plus tactile stimulators (24.35 ± 13.06 cm). Compared with no stimulators, net tarsal energy generation increased for tactile stimulators, limb weights, and limb weights plus tactile stimulators, but only the weighted conditions increased net energy generation across the hip joint. Conclusions and Clinical Relevance—The type and weight of foot stimulators affected the magnitude of the kinematic and kinetic responses and the joints affected. These findings suggest that different types of foot stimulators are appropriate for rehabilitation of specific hind limb gait deficits, such as toe dragging and a short stride.