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.     

Association between in-transit loss, internal trailer temperature, and distance traveled by Ontario market hogs

An observational study was conducted from July to October 2004 to determine the association between in-transit losses of swine and internal trailer temperature after controlling for loading density, trip distance, herd size, and random trip effect. A convenience sample of 3 trucking companies was used to collect temperature, relative humidity, and global positioning data for 104 trips that delivered 21 834 pigs from 371 producers to Ontario abattoirs. The association between in-transit loss and trailer temperature was determined using the 90th percentiles of internal temperature for each trip. Average loading density was 0.36 m²/100 kg pig (range 0.28 to 0.50 m²/100 kg pig). Average in-transit loss was 0.12%; however, 94% of producers experienced no losses. As the 90th percentile of internal trailer temperature increased from a range of 8.6°C to 23.3°C to a range of 23.4°C to 26.1°C, average in-transit loss ratio increased approximately 3-fold, with an additional 2-fold increase as the range increased from 26.2°C to 28.9°C to 29.0°C to 30.5°C. As the 90th percentile of temperature increased by 1°C over the full range of temperatures in this study, in-transit loss was expected to increase 1.26 times. The in-transit loss was expected to decrease 0.81 times for each 50-km increase in distance traveled between the farm and the abattoir.     

Subacute atropine toxicity in a pygmy sperm whale, Kogia breviceps.

Atropine, an anticholinergic agent commonly used in human and veterinary medicine, is reported to cause toxicity associated with its antimuscarinic action. A juvenile pygmy sperm whale, Kogia breviceps, was treated with atropine in an attempt to relieve symptoms similar to pyloric stenosis, as has been used in humans. Two doses of 0.01 mg/kg were given i.m., 12 hr apart, followed by three doses of 0.005 mg/kg i.m. s.i.d. over the next 3 days. Symptoms associated with atropine toxicity developed gradually and included hyperexcitability, a generalized ascending paralysis of body musculature, shallow, rapid respiration, vomiting, aspiration of seawater, and pulmonary edema. Treatment with physostigmine salicylate (two doses of 2 mg i.m., I hr apart) was effective in counteracting the paralysis, as well as other symptoms, beginning in as little as 17 min after the first dose, and the whale was back to swimming on its own after 8 hr. All overt symptoms of atropine toxicity were gone in about 24 hr, but there were other possible sequella that lasted much longer.     

Detomidine-Propofol Anesthesia for Abdominal Surgery in Horses

OBJECTIVE: To evaluate propofol for induction and maintenance of anesthesia, after detomidine premedication, in horses undergoing abdominal surgery for creation of an experimental intestinal adhesion model. STUDY DESIGN: Prospective study. ANIMALS: Twelve horses (424 +/- 81 kg) from 1 to 20 years of age (5 females, 7 males). METHODS: Horses were premedicated with detomidine (0.015 mg/kg i.v.) 20 to 25 minutes before induction, and a propofol bolus (2 mg/kg i.v.) was administered for induction. Propofol infusion (0.2 mg/kg/min i.v.) was used to maintain anesthesia. The infusion rate was adjusted to maintain an acceptable anesthetic plane as determined by muscle relaxation, occular signs, response to surgery, and cardiopulmonary responses. Oxygen (15 L/min) was insufflated through an endotracheal tube as necessary to maintain the SpO2 greater than 90%. Systolic (SAP), mean (MAP), and diastolic (DAP) arterial pressures, heart rate (HR), electrocardiogram (ECG), respiratory rate (RR), SpO2 (via pulse oximetry), and nasal temperature were recorded at 15 minute intervals, before premedication and after induction of anesthesia. Arterial blood gas samples were collected at the same times. Objective data are reported as mean (+/-SD); subjective data are reported as medians (range). RESULTS: Propofol (2.0 mg/kg i.v.) induced anesthesia (mean bolus time, 85 sec) within 24 sec (+/-22 sec) after the bolus was completed. Induction was good in 10 horses; 2 horses showed signs of excitement and these two inductions were not smooth. Propofol infusion (0.18 mg/kg/min +/- 0.04) was used to maintain anesthesia for 61 +/- 19 minutes with the horses in dorsal recumbency. Mean SAP, DAP, and MAP increased significantly over time from 131 to 148, 89 to 101, and 105 to 121 mm Hg, respectively. Mean HR varied over time from 43 to 45 beats/min, whereas mean RR increased significantly over anesthesia time from 4 to 6 breaths/min. Mean arterial pH decreased from a baseline of 7.41 +/- 0.07 to 7.30 +/- 0.05 at 15 minutes of anesthesia, then increased towards baseline values. Mean PaCO2 values increased during anesthesia, ranging from 47 to 61 mm Hg whereas PaO2 values decreased from baseline (97 +/- 20 mm Hg), ranging from 42 to 57 mm Hg. Muscle relaxation was good and no horses moved during surgery: Recovery was good in 9 horses and acceptable in 3; mean recovery time was 67 +/- 29 minutes with 2.4 +/- 2.4 attempts necessary for the horses to stand. CONCLUSIONS: Detomidine-propofol anesthesia in horses in dorsal recumbency was associated with little cardiovascular depression, but hypoxemia and respiratory depression occurred and some excitement was seen on induction. CLINICAL RELEVANCE: Detomidine-propofol anesthesia is not recommended for surgical procedures in horses if dorsal recumbency is necessary and supplemental oxygen is not available (eg, field anesthesia).     

Participatory plant breeding approach for host plant resistance to bean fly in common bean under semi-arid Kenya conditions

Common bean (Phaseolus vulgaris L.) is the most important legume crop in Kenya. It is cultivated across a wide range of agro-ecologies which include high potential and marginal areas. Eastern Kenya alone, largely semi-arid, accounts for 35% of the country’s total bean production. Bean farmers mainly small-scale have limited access to quality seed, chemical pesticides and fertilizer. Therefore, bean yield under on-farm conditions still remains below 500 kg ha⁻¹ while the potential is about 1,200 kg ha⁻¹ under semi-arid conditions. To asses the farmers’ views on bean varieties and a key insect pest and associated constraints contributing to yield loss, research was undertaken. The research included survey to quantify the yield loss and Participatory Rural Appraisal to determine the level of adoption and criteria for variety choice in semi-arid eastern Kenya (SAEK). The results show that farmers consider drought and insect pest problems as main causes for low yields. The adoption rate for improved varieties is high but self sufficiency in beans stands at 23% in the dry transitional (DT) agro-ecology and at 18% in the dry mid-altitude (DM) agro-ecology, respectively. This could be attributed to low adaptability since most of the improved varieties grown were selected for high potential areas but now found in marginal areas. Drought, earliness, yield stability, and insect pest resistance are the main reasons for choice of varieties by farmers. Bean fly (Ophiomyia sp.) was identified as one of the key crop pests of beans limiting yield. Besides, African bollworm (Helicoverpa armigera) and bean aphid (Aphis fabae) were ranked higher. Due to limitations of the conventional breeding approach, a participatory plant breeding approach is suggested so as to provide an opportunity to develop insect pest resistant varieties adapted to the SAEK region.     

Evaluation of a radioimmunoassay for type III procollagen peptide in the dog

Type III procollagen peptide (P-3-P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P-3-P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P-3-P, followed by Western immunoblotting with rabbit aniti-P-3-P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P-3-P and sera from two dogs suspected of having elevated P-3-P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross-reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P-3-P. All tested sera from dogs had minimal competitive binding with radiolabeled P-3-P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P-3-P concentrations in any of the groups of dogs studied. It was concluded that currently available radioimmunoassay kits for the measurement of P-3-P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P-3-P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.     

HYPERTROPHIC OSTEOPATHY ASSOCIATED WITH PULMONARY BLASTOMYCOSIS IN A DOG

Hypertrophic osteopathy (HO) associated with pulmonary blastomycosis was diagnosed in a 5–year-old male mixed-breed dog. One year prior to referral, increased pulmonary opacity had been identified on radiographs made during an examination for a chronic cough. Although serologic tests for blastomycosis were negative, the dog was treated with oral ketoconozole on the basis of suspicious lesions seen on radiographs and clinical signs. Ten months after completing the ketoconozole therapy, the dog was presented for a persistent cough and lameness. Intrapulmonary masses and periosteat proliferation were observed radiographically. A biopsy of the pulmonary masses was done, and Blastomyces dermatitidis was identified. Amphotericin B and ketoconozole administration resulted in clinical improvement and partial resolution of the HO lesions five months after initiation of therapy.     

Mechanisms conferring resistance of Western flower thrips to bendiocarb

Mechanisms associated with bendiocarb resistance were examined in two strains of western flower thrips, Frankliniella occidentalis (Pergande), that differed in their susceptibility to this carbamate by 13.6-fold. No appreciable differences between the two strains in [¹⁴C] bendiocarb penetration and excretion were detected; however, bendiocarb was metabolised substantially faster by the resistant KCM thrips than by the more susceptible UMC thrips. No appreciable difference between the two strains was found in the sensitivity of acetylcholinesterase activity to inhibition by bendiocarb. It was concluded that bendiocarb resistance in KCM western flower thrips was due to enhanced metabolism that probably was mainly oxidative in nature.