Development of an assay to determine single nucleotide polymorphisms in the prion gene for the genetic diagnosis of relative susceptibility to classical scrapie in sheep.

The objective of this study was to develop a reliable Taqman 5' Nuclease Assay for genotyping sheep for scrapie susceptibility. The sheep prion gene contains 2 single nucleotide polymorphisms (SNPs) that may mediate resistance to classical scrapie, one at codon 136, alanine (A) or valine (V), and another at codon 171, arginine (R) or glutamine (Q). The R allele appears to confer resistance to classical scrapie, with the AA(136) RR(171) genotype the most resistant to scrapie and QR(171) only rarely infected in the US sheep population. The Assays by Design protocol was used for development of probes and primers for codon 136 and Primer Express for codon 171. Commercially available kits were used to isolate genomic DNA from blood or muscle. For validation, 70 SNP determinations for each codon were compared to commercial testing with an error rate of less than 1%. Then, 935 samples from blood (n = 818) and muscle (n = 117) were tested for both codons with 928 successful determinations and only 7 samples (<1% of total samples) that needed repeating. Genotypes were AA QQ (n = 102; 11.0%), AV QQ (n = 28; 3.0%), AA QR (n = 396; 42.7%), AV QR (n = 54; 5.8%), and AA RR (n = 348; 37.5%). Thus, 86% of the sheep tested (n = 798) contained R at codon 171 and were expected to be scrapie-resistant. This new Taqman 5' Nuclease SNP genotyping assay is accurate, easy to perform, and useful in the study of classical scrapie in sheep and its prevention through selective breeding programs to eliminate highly susceptible animals.     

Microchip-associated leiomyosarcoma in an Egyptian fruit bat (Rousettus aegyptiacus).

Microchips are commonly used in companion, research, and zoo animal medicine for easy, reliable, and cost-effective identification with relatively low risk of side effects. This report describes development of a metastatic leiomyosarcoma associated with a microchip in an Egyptian fruit bat (Rousettus aegyptiacus).     

Hyperthermia and Delayed-Onset Myopathy after Recovery from Anesthesia in a Horse

A 13-year old Thoroughbred cross gelding (528 kg) underwent lameness investigation. Because of his temperament, general anesthesia was required to facilitate ultrasound of the left fore fetlock and intra-articular medication of three joints. Anesthesia was induced with ketamine/diazepam after acepromazine/detomidine premedication. Anesthesia was maintained for 40 minutes with a guaifenesin/detomidine/ketamine intravenous infusion. Recovery from anesthesia was initially uneventful, although of a moderate duration (70 minutes). Once standing, the horse proceeded to box walk in an agitated state and became recumbent on two occasions. The horse was manually restrained, at which time its rectal temperature was 41.8°C. Cooling measures were employed (fans, ice-water enemas, wet rugs, intravenous fluid therapy (IVFT), and topical application of surgical spirit) until rectal temperature reached 38.7°C. IVFT was continued for a further 16 hours. Four days after recovery from anesthesia, bilateral triceps, deltoideus, trapezius, and rhomboideus muscle swelling was observed. Blood creatinine kinase was elevated (24,898 IU/L). Treatment for postanesthetic myopathy was initiated (hot packing of the muscle groups, topical dimethylsulfoxide [DMSO] cream application, and oral phenylbutazone). Myoglobinuria was not observed at any time. Muscle swelling decreased over the following 3 days. The horse was discharged on day 11 and has since returned to work.     

Relative Flow-Time Relationships in Single Breaths Recorded After Treadmill Exercise in Thoroughbred Horses

Analysis of individual breaths after exercise has potential for pulmonary function testing. The aims of this study were to investigate the dependence of measurements of pulmonary function in single breaths on time postexercise and tidal volume (V_T) after treadmill exercise. Five Thoroughbred horses without evidence of airway disease were used. Horses had been previously acclimated to treadmill exercise and to wearing a face mask. A Quadflow spirometer recorded airflow rates continuously during 90 seconds after intense treadmill exercise to fatigue. Indices of function were based on ratios of times within each breath and analyses of the shape of relative flow–time curves within inspiration and expiration. Restricted maximum likelihood, general linear regression, repeated-measures one-way analysis of variance (ANOVA), and two sample t-tests were used, with statistical significance at P < .05. Time postexercise had no effect on several ratios based on time for inspiration (T_I) and expiration (T_E), and times to peak flows (T_I/T_T, T_E/T_T, T_E/T_I, Tpef [peak expiratory flow]/T_E, and Tpif [peak inspiratory flow]/T_I). Many variables were significantly dependent on V_T. Occasional big respiratory cycles with V_T more than 10% greater than in the previous breath had significantly different means for relative flow (Rf)/(T_E/T_I), epz_50% (50% of the time from Tpef to end of expiration), epz_75% (75% of the time from Tpef to end of expiration), and ipz_75% (75% of the time from Tpif to end of inspiration). Predicted means for these variables differed by 10–20%. This study establishes guidelines for the selection of breaths after exercise, and describes a new approach to measurement of relative flow and time relationships. It was concluded that several time-based ratios have potential for measuring pulmonary function. However, care is needed when selecting breaths for calculation of most of the new relative flow–time variables.     

Associations between yearling body measurements and career racing performance in Thoroughbred racehorses

Yearling morphometric measurements and variables related to career racing performance were collected on 260 Thoroughbreds to determine correlations between and among these traits. Morphometric measurements were adjusted to 365 days based on interpolation from adjacent age points and included wither and hip height, body length, distal limb length, cannon circumference, heartgirth circumference, and chest width. Measures of career racing ability included total earnings, win percentage, and Standard Starts Index (SSI). Wither height and hip height were favorably correlated with lifetime earnings, SSI, and win percentage. Body length and heartgirth were positively correlated with SSI and win percentage. Leg traits were not correlated with any performance parameters. Horses that had placed in or won a stakes race tended to be taller at the wither and hip as yearlings. Horses categorized as preferring turf tended to be taller at the wither and hip at 365 days than those that preferred to run on dirt. No significant difference in yearling morphometric measurements between sprinters and routers was detected. In addition to phenotype and pedigree information, morphometric measurements taken in the yearling year may allow for more accurate estimation of future performance in the Thoroughbred racehorse.     

Comparison of Plasma Fentanyl Concentrations by Using Three Transdermal Fentanyl Patch Sizes in Dogs

Objective—To compare plasma fentanyl concentrations attained after the application of three transdermal fentanyl patch sizes (50, 75, and 100 μg/hour) in dogs. Design—Repeated Latin square controlled study. Animals—Six intact, mixed-breed adult dogs (2 males, 4 females) weighing 19.9 ± 3.4 kg. Methods—Each dog was randomly assigned to receive each of three treatments: 50 (P50), 75 (P75), or 100 (P100) μg/hour transdermal patches. Patches were left in place for 72 hours. Jugular venous blood was collected at 1,2, 4, 8, 12, 24, 36, 48, 60, and 72 hours after patch application and for 1, 2, 4, 8, and 12 hours after patch removal. Plasma fentanyl concentrations were measured using a radioimmunoassay technique. After a 96-hour washout period, each dog was moved to another treatment group and received a different patch size. Results—The following results were obtained (mean ± SD): average plasma fentanyl concentration from 24 to 72 hours, 0.7 ± 0.2 ng/mL (P50), 1.4 ± 0.5 ng/mL (P75), 1.2 ± 0.5 ng/mL (P100); the total area under the concentration versus time curve (0 hours to infinity), 46 ± 12.2 ng/h/mL (P50), 101.2 ± 41.4 ng/h/mL (P75), 80.4 ± 38.3 ng/h/mL (P100); and the apparent elimination half-life, 3.6 ± 1.2 hours (P50), 3.4 ± 2.7 hours (P75), and 2.5 ± 2.0 hours (P100). There was a high degree of variability in plasma fentanyl concentrations achieved. Plasma fentanyl concentrations declined rapidly after patch removal. Conclusions—The attainment of steady-state plasma concentrations takes up to 24 hours, and there is a great deal of variability in the final concentrations reached in different individuals. In this study, the 100 μg/hour patches did not provide statistically increased plasma concentrations when compared with the 50 μg/hour patches. Clinical Relevance—Because of the interindividual and intraindividual variation in plasma fentanyl concentrations, patches should be applied 24 hours before the anticipated time that analgesia will be required. Adequacy of analgesia and potentially deleterious side effects, such as sedation and respiratory depression, should be monitored while the patches are in place. Skin reactions may occur, and the patches should be removed if such skin irritation is seen. After the patch is removed, it is expected that analgesia will wane rapidly because of the brief elimination half-life.     

DETECTION OF PORTAL BLOOD FLOW USING PER-RECTAL 99mTc-PERTECHNETATE SCINTIGRAPHY IN NORMAL CATS

This study reports data obtained from per-rectal ^99mTc-pertechnetate portal scintigraphy in normal cats. It examines the effects of chemical restraint and the methods employed in defining regions of interest (ROIs) on the shunt index derived from this data. Six normal cats were used for the study; all six were chemically restrained for imaging using propofol and later four of them were manually restrained for comparison. Portal blood flow was studied and the mean shunt index was found to be 5.9%± 3.9 when ROIs were operator defined and 9.2%± 4.4 when ROIs were defined using an isocontour program. In cats that were restrained using propofol and operator defined ROIs, the mean value for the time between detection of radioactivity in the liver and in the heart was 14 ± 1 seconds.     

Esophageal Replacement in the Dog by Microvascular Colon Transfer

An orthotopic colon graft based on the middle colic artery and vein was implanted with microvascular technique and a stapling instrument in five dogs. The grafts were successful in four dogs. A similar colon autograft was used to replace the entire thoracic esophagus in five dogs. The recipient vessels were the left carotid artery and left external jugular vein. Four of the grafts failed because of kinking and thrombosis of the arterial supply (2 dogs) or the venous outflow (2 dogs). One graft, which had a viable vascular supply, developed a severe leak at the colon-to-stomach anastomosis, and the dog was euthanatized on day 3. The recipient vascular pedicle was modified and used successfully to replace a portion of the cervical esophagus in three dogs. The grafts survived, the dogs could swallow liquids and semisolid food well, and, at necropsy after 4 weeks, the anastomotic sites were well healed. The graft sites contained essentially normal colon mucosa.     

Holding Power of Orthopedic Screws in Equine Third Metacarpal and Metatarsal Bones: Part II. Adult Horse Bone

Comparison was made of the holding power of 5.5 and 4.5 mm cortical orthopedic screws inserted into third metacarpal and metatarsal cadaver bones from 3- and 8-year-old horses. The tensile strength of these screws was tested mechanically. In nine comparative trials of these screws, 5.5 mm screws pulled out of bone in five trials at an average of 116.0 kg tensile force and broke in four trials at an average of 1383.2 kg. A 4.5 mm screw pulled out of bone at 834.5 kg in one trial, and screws broke at an average of 849.2 kg in eight trials. The larger 5.5 mm screw required a significantly greater (p = 0.022) pullout force than the mean force at 4.5 mm screw breakage. Fixation failure was due to screw breakage or bone shear, with 5.5 mm screws occasionally creating bone fragmentation during pullout. The average tensile breaking strengths of the 5.5 mm screws (1391.4 kg) and 4.5 mm screws (832.7 kg) determined mechanically were similar to forces at screw breakage during pullout testing in bone. Since the 5.5 mm screws have greater holding power and tensile strength than 4.5 mm screws, the use of the 5.5 mm screw in fracture repair in adult horses is recommended.     

Variation in crushing strength and some related properties of a red pine

Summary Below-crown radial and height variations in maximum crushing strength parallel to grain, relative density and tracheid length of an immature, 55 year old, red pine were studied. All properties showed pronounced trends with radial age and increased from pith to bark. A radial age of 16 years was estimated as the age from which mature stem wood is formed. All properties studied varied with height above ground. Strong correlations were observed among properties. The study tree exhibited noticeably lower average crushing strength than is reported in the literature for wood from mature red pines.This work was undertaken with financial support from Karadeniz Technical University, Trabzon, Turkey, and the Natural Sciences and Engineering Research Council of Canada, Operating Grant 0GP0004417