Design and evaluation of primer pairs for efficient detection of avian rotavirus

The use of molecular methods for rotavirus characterisation provides increased sensitivity for typing and allows the identification of putative reassortant strains. Reagents and methods for genotyping the virus need constant modification because of the reassortant nature of the virus. This study was aimed at designing and evaluating new oligonucleotide degenerate primer pairs that provide increased sensitivity and specificity for detecting avian rotavirus. Gene-specific primer pairs were designed by analysing different rotavirus strains isolated during the last decade by downloading them from the GenBank. The alignments were generated using clustal analysis from the BioEdit program. Degenerate nucleotides were included due to the reassortant nature of rotavirus. The consensus sequences were aligned using the BioEdit program and then treated with the Fast PCR software to derive the primers. The derived primer sequences were submitted for a BLAST search to ensure alignment was exclusive to the desired target genes. The designed primers had specific bands and were efficient in detecting rotavirus in faecal samples than previously published primers. Thus, a successful surveillance of rotaviruses requires that primer pairs be updated regularly in order to detect the emergence of novel or “unusual types”, which have occurred by genetic drift causing nucleotide changes at the primer binding sites that result in typing failures. We recommend the use of the proposed primers in molecular surveillance studies for efficient detection of avian rotavirus.     

Alanine aminotransferase apoenzyme in dogs

Unusually low serum activity of alanine aminotransferase (ALT) was detected in a Rottweiler dog with gastric dilatation-volvulus. Activity of ALT in the same sample was found to be much higher (estimated increase of 14,225%) when measured by methods adding the cofactor pyridoxal-5'-phosphate (P5P), indicating that nearly all serum ALT was in the apoenzyme form. An investigation was undertaken to determine the frequency of high serum aminotransferase apoenzyme levels in dogs. Eighty canine serum samples submitted to the Clinical Pathology Laboratory at Cornell University were assayed for ALT and aspartate aminotransferase (AST) in the presence or absence of exogenous P5P. In 79 dogs, inclusion of P5P in the LAT assay resulted in a median decrease in AST activity of -6.3% (range -33.3% to 25.0%) in all 80 dogs. One dog had an increase of 336% in ALT activity after inclusion of P5P in the assay, but lacked a similar increase in AST activity. The reason for the high levels of ALT apoenzyme in the 2 dogs was not determined.     

Pre-operative fibrous osteodystrophy and severe,refractory, post-operative hypocalcemia following parathyroidectomy in a dog

A 13-year-old dog exhibited dramatic, radiographic osteopenia consistent with fibrous osteodystrophy secondary to primary hyperparathyroidism. Following parathyroidectomy, the dog developed severe, prolonged hypocalcemia, but was successfully treated and discharged 32 d after surgery. A variety of factors may have contributed to this dog’s hypocalcemia including hypoparathyroidism and hungry bone syndrome.     

Population pharmacokinetics of methadone hydrochloride after a single intramuscular administration in adult Japanese sika deer (Cervus nippon nippon)

ObjectiveTo assess the population pharmacokinetics of methadone in deer.Study designProspective non-randomized experimental trial.AnimalsTwelve healthy adult sika deer (nine males and three females).MethodsDeer received intramuscular administration of racemic methadone hydrochloride at 0.5 mg kg⁻¹ or 1 mg kg⁻¹. Plasma methadone and its metabolite 2-Ethylidene-1,5-Dimethyl-3,3-Diphenyl-Pyrolidine (EDDP) concentrations were determined by validated liquid chromatography coupled to tandem mass spectrometry methods, at times 0, 30 minutes, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours. Population pharmacokinetics analysis was undertaken using a non-linear mixed effects modelling (NONMEM).ResultsA two-compartment linear disposition model best described observed time-concentration profiles of methadone and EDDP. Population parameter estimates of methadone were elimination clearance (17.3 L hour⁻¹), metabolic clearance (34.6 L hour⁻¹), volume of distribution of compartment 1 (216.0 L) and volume of distribution of compartment 2 (384.0 L). Population parameter estimates of EDDP were elimination clearance (121.0 L hour⁻¹), volume of distribution of compartment 3 (1.08 L) and volume of distribution of compartment 4 (499.5 L). The total clearance and total volume of distribution of methadone and EDDP were 51.9 L hour⁻¹, 121.0 L hour ⁻¹, 600.0 L and 500.6 L, respectively. The methadone terminal elimination half-life was 8.19 hours. No adverse effects were observed after methadone administration.Conclusions and Clinical relevanceFollowing intramuscular injection, methadone was characterized by a large total volume of distribution, high systemic clearance and intermediate terminal half-life in sika deer.     

Effects of two different anesthetic protocols on 64‐MDCT coronary angiography in dogs

Heart rate is a major factor influencing diagnostic image quality in computed tomographic coronary artery angiography (MDCT‐CA), with an ideal heart rate of 60–65 beats/min in humans. The purpose of this prospective study was to compare effects of two different clinically applicable anesthetic protocols on cardiovascular parameters and 64‐MDCT‐CA quality in 10 healthy dogs. Scan protocols and bolus volumes were standardized. Image evaluations were performed in random order by a board‐certified veterinary radiologist who was unaware of anesthetic protocols used. Heart rate during image acquisition did not differ between protocols (P = 1), with 80.6 ± 7.5 bpm for protocol A and 79.2 ± 14.2 bpm for protocol B. Mean blood pressure was significantly higher (P > 0.05) using protocol B (protocol A 62.9 ± 9.1 vs. protocol B 72.4 ± 15.9 mmHg). The R‐R intervals allowing for best depiction of individual coronary artery segments were found in the end diastolic period and varied between the 70% and 95% interval. Diagnostic quality was rated excellent, good, and moderate in the majority of the segments evaluated, with higher scores given for more proximal segments and lower for more distal segments, respectively. Blur was the most commonly observed artifact and mainly affected the distal segments. No significant differences were identified between the two protocols for optimal reconstruction interval, diagnostic quality and measured length individual segments, or proximal diameter of the coronary arteries (P = 1). Findings indicated that, when used with a standardized bolus volume, both of these anesthetic protocols yielded diagnostic quality coronary 64‐MDCT‐CA exams in healthy dogs.     

Multicenter, randomized controlled trial of pain-related behaviors following routine neutering in dogs

OBJECTIVE: To evaluate the degree of postoperative pain in dogs undergoing elective castration or ovariohysterectomy (OHE); determine whether an association exists between surgeon experience, incision length, or surgery duration and degree of postoperative pain; and determine whether analgesic treatment decreases expression of postoperative pain behaviors. DESIGN: Randomized controlled clinical trial. ANIMALS: 426 client-owned dogs undergoing OHE or castration. PROCEDURES: Dogs underwent OHE or castration performed by an experienced veterinarian or a fourth-year veterinary student. Dogs were randomly assigned to 1 of 4 treatment groups: no perioperative analgesic treatment (n = 44), preoperative administration of morphine (144), preoperative administration of nalbuphine (119), and postoperative administration of ketoprofen (119). Dogs were evaluated while in the hospital before anesthesia and for 4 hours after surgery and once a day at home for 3 days after surgery. RESULTS: Dogs in all 4 groups had significant increases in overall pain scores after surgery, compared with baseline scores. There were significant differences among groups, with control dogs having significantly higher increases in overall pain scores than dogs in the other groups. Factors that did not influence the frequency or severity of pain-related behaviors included breed, individual hospital, anesthetic induction protocol, surgeon experience, and duration of surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that dogs expressed behaviors suggestive of pain following OHE and castration, that analgesic treatment mitigated the expression of pain-related behaviors, and that surgeon experience and surgery duration did not have any effect on expression of pain-related behaviors.     

COMPARATIVE ORGAN IMAGING: THE GASTROINTESTINAL TRACT

Diagnostic radiology of the esophagus, stomach, small and large intestine, mesentery, and pancreas is reviewed. Survey and contrast radiography remain the principal imaging methods used to evaluate gastrointestinal diseases in animals. Of the other imaging techniques, scintigraphy has potential in assessment of esophageal and gastric motility disorders, and detection of intestinal hemorrhage. Ultrasonography is useful for imaging the pancreas and gastrointestinal mass lesions.     

Evaluation of the safety of ivermectin-praziquantel administered orally to pregnant mares

OBJECTIVE: To evaluate the safety of an orally administered ivermectin and praziquantel paste with regard to variables associated with clinical findings, parturition, lactation, maternal care, and neonate viability in pregnant mares. ANIMALS: 40 pregnant mares. PROCEDURE: Mares were randomly allocated into treatment (n = 20) and control (20) groups and administered a placebo or 3 times the therapeutic dosage of ivermectin (0.6 mg/kg) and praziquantel (4.5 mg/kg) at 14-day intervals until parturition. Physical examinations were performed on mares and their foals after parturition (on postpartum days 30, 60, and 90) to identify any drug-related effects. As an aid in assessing general health, hematologic and serum biochemical analyses were performed monthly on the mares. RESULTS: In blood constituents, minor alterations that were not biologically important were observed. Reproductive performance was not affected by the unusual treatment duration or high dosage, although the drugs were administered during a crucial period of equine embryonic development (30 to 60 days). Neither adverse effects on mares nor abortions occurred. Follow-up evaluations of the foals for a 3-month period did not detect any abnormalities. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of the ivermectin-praziquantel paste appears to be safe in pregnant mares and their foals.