Use of a flow-mediated vasodilation technique to assess endothelial function in dogs

OBJECTIVE: To develop and assess the reproducibility of a protocol to noninvasively test endothelial function in dogs on the basis of the flow-mediated vasodilation (FMD) procedure used in humans. ANIMALS: 5 healthy spayed female dogs. PROCEDURES: Luminal arterial diameter and blood flow velocity in the brachial and femoral arteries were measured with ultrasonography. The within-dog reproducibility of these ultrasonographic measurements was tested. An occlusion period of 1, 3, or 5 minutes with an inflatable cuff was used to create the FMD response. Measurements made at 15, 30, and 60 seconds following release of the occlusion were compared with measurements made immediately prior to each occlusion to assess the FMD response. RESULTS: Within-dog reproducibility of measurements revealed moderate to high correlations. Change from baseline in luminal arterial diameter was most substantial when measured at 30 seconds following release of occlusion, whereas blood flow velocity changes were maximal when measured at 15 seconds following release. The brachial imaging site provided a larger number of significant FMD responses than the femoral site. The 3-minute occlusion period provided equal or better responses than the 5-minute occlusion period. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasonographic measurement of the FMD responses was a feasible and reproducible technique and significant changes from baseline were detected. The FMD responses in dogs were most substantial when performed at the brachial artery with blood flow velocity and luminal arterial diameter changes from baseline measured at 15 and 30 seconds, respectively, following release of a 3-minute occlusion period.     

Identification and Quantification of Pathogenic Pythium spp. from Soils in Eastern Washington Using Real-Time Polymerase Chain Reaction

ABSTRACT Traditional methods of quantifying Pythium spp. rely on the use of selective media and dilution plating. However, high variability is inherent in this type of enumeration and counts may not be representative of the pathogenic population of Pythium spp. Variable regions of the internal transcribed spacer of the rDNA were used to design species-specific primers for detection and quantification of nine Pythium spp. from soils in eastern Washington. Primer pairs were designed for Pythium abappressorium, P. attrantheridium, P. heterothallicum, P. irregulare group I, P. irregulare group IV, P. paroecandrum, P. rostratifingens, P. sylvaticum, and P. ultimum and used with real-time polymerase chain reaction. Standard curves were generated for each of the species using SYBR Green I fluorescent dye for detection of amplification. Seventy-seven isolates of Pythium were screened to confirm specificity of each primer set. DNA was extracted from soil and standard curves were generated for P. irregulare group I, P. irregulare group IV, and P. ultimum to correlate populations of each species in the soil with quantities of DNA amplified from the same soil. Examination of raw field soils revealed results similar to those observed in previous studies. This new technique for the quantification of Pythium spp. is rapid and accurate, and will be a useful tool in the future study of these pathogenic Pythium spp.     

Effect of cereal type and exogenous enzyme supplementation in pig diets on odour and ammonia emissions

A 2 × 2 factorial experiment was conducted to investigate the interaction between cereal type (wheat vs. barley) and exogenous enzyme supplementation (with or without) on odour concentration and ammonia emissions in grower–finisher pigs. The enzyme supplement used contained Endo-1, 3 (4)-β-glucanase (EC 3.2.1.6) and Endo-1, 4-β-xylanase (E.C 3.2.1.8). The diets were formulated to contain similar levels of digestible energy and lysine. The experimental treatments were as follows: (1) wheat-based diet, (2) wheat-based diet containing a β-glucanase and β-xylanase mixed enzyme supplement, (3) barley-based diet and (4) barley-based diet containing a β-glucanase and β-xylanase mixed enzyme supplement. The diets were offered to the pigs for 4 weeks and this was repeated 4 times (n = 4). Odour and ammonia emissions were measured on days 9, 11, 14, 16, 21 and 23 of each experimental period. Odour samples were collected in 20-litre Nalophan bags and analysed for odour concentration using an ECOMA Yes/No Olfactometer. Ammonia concentrations were measured using Dräger tubes. There was a cereal × enzyme interaction in odour emission rates (P < 0.05) and in ammonia emissions (P < 0.01). The addition of an enzyme supplement increased odour emissions and had no effect on ammonia emissions in the barley-based diet. The enzyme supplementation had no effect on odour but decreased ammonia emissions in the wheat based diet.