Standardisation of a coproantigen reduction test (CRT) protocol for the diagnosis of resistance to triclabendazole in Fasciola hepatica

A sheep trial was performed to standardise a coproantigen reduction test (CRT) protocol for the diagnosis of resistance to triclabendazole (TCBZ) in Fasciola hepatica). The CRT employs the BIO K201 Fasciola coproantigen ELISA (Bio-X Diagnostics, Jemelle, Belgium) to test for the presence of F. hepatica coproantigens in a faecal sample. If it is coproantigen-positive, the CRT protocol recommends that faecal samples are re-tested for coproantigens at 14 days post-treatment (dpt), with negative testing at this point indicating TCBZ success. Initial work aimed to confirm the sensitivity of the BIO K201 ELISA for Fasciola infection and investigate whether coproantigens represent a robust reduction marker of TCBZ efficacy. Thirty-eight, indoor-reared sheep were artificially infected with F. hepatica isolates known to be susceptible (Cullompton) and resistant (Sligo) to TCBZ action, respectively. Treatment was administered at 12 weeks post-infection (wpi), with 2 sheep groups, infected with each isolate, culled at 2 and 4 weeks post-treatment (wpt), respectively. Necropsy was performed to confirm treatment efficacy. Individual faecal samples were collected twice-weekly throughout the trial period. Additional work focused on the effect of temperature on faecal sample collection and storage. Faecal samples collected from sheep positive for F. hepatica infection were sub-sampled and left at room temperature. Individual sub-samples were tested by ELISA on consecutive days and these readings compared to the original test result on the day of collection. In addition, ELISA values were compared between faecal sub-samples prepared on the day of sampling and post storage at -20°C. Also, an immunocytochemical study was performed to determine the tissue site of origin of the coproantigen protein in the fluke. Results showed that the BIO K201 ELISA was sensitive for Fasciola coproantigens, with coproantigens detectable from 5 wpi onwards. The suitability of coproantigens as a diagnostic marker of TCBZ efficacy was supported by the absence and presence of coproantigens in TCBZ-treated Cullompton (TCBZ-susceptible) and Sligo (TCBZ-resistant) F. hepatica infections at 2 and 4 wpt, respectively. Study results suggest that low to moderate temperature has little, if any, impact on coproantigen stability in faecal samples, but that higher temperatures may have. Immunolabelling for the coproantigen showed that it was specific to the gastrodermal cells of both adult and juvenile flukes.     

POISONING OF CHICKENS AND DUCKS BY PYRROLIZIDINE ALKALOIDS OF HELIOTROPIUM EUROPAEUM

The disease produced by feeding chickens and ducks a commercial poultry feed containing heliotrine and lasiocarpine, pyrrolizidine alkaloids of Heliotropium europaeum, is described. Illthrift, ascites and degenerative lesions in the liver were the major findings. Similar lesions occurred in chickens fed a diet containing H. europaeum. The source of the alkaloids in commercial poultry feed was probably the seeds of H. europaeum harvested with wheat.     

Demonstration of regional differences in equine ventricular myocardial velocity in normal 2-year-old Thoroughbreds with Doppler tissue imaging

REASONS FOR PERFORMING STUDY: Doppler tissue imaging (DTI) is a novel noninvasive method by which myocardial velocity can be assessed directly and it allows regional, rather than global, cardiac function to be evaluated. HYPOTHESIS: That regional differences in myocardial velocities exist within the equine ventricle. OBJECTIVES: To develop a repeatable examination technique for DTI in horses, describe DTI findings in various regions of the normal equine ventricle, compare colour (CDTI) and spectral (SDTI) techniques of DTI, and document regional differences in myocardial velocity. METHODS: Five regions of the ventricles (right ventricular wall, interventricular septum and left, right and caudal regions of the left ventricle) were evaluated using SDTI and CDTI in 20 clinically normal Thoroughbreds age 2 years. Individual repeatability of the method was determined by examination of one 6-year-old Thoroughbred on 6 occasions. RESULTS: Three major movements were observed in the ventricular walls in systole, early diastole and late diastole. The interventricular septum had a complex pattern of movement. The left region of the left ventricle and interventricular septum had the most rapid movement. The individual repeatability of CDTI was poor, while in systole and early diastole, but not late diastole, SDTI produced repeatable estimates of maximal myocardial velocity. The different velocity estimates obtained with SDTI and CDTI are not interchangeable. Regional differences in the peak mean and maximal myocardial velocities were found in systole and early diastole (P<0.05), but were not identified in late diastole. CONCLUSIONS: The SDTI modality appears to produce the most repeatable data. There are regional differences in myocardial velocity within the equine ventricles for systole and early diastole. POTENTIAL RELEVANCE: DTI shows potential as a tool for studying regional myocardial movement both in clinical cases suspected of having myocardial dysfunction and in a research setting. In particular, SDTI offers potential as a direct and noninvasive means to study early diastolic function of the equine ventricles.     

Influence of temperature stress on the energy and protein metabolism and requirements of the working horse

In coping with the effects of high temperatures, the movement of performance horses from temperate to tropical and sub-tropical regions places major demands on their animals metabolic and cooling systems. Without acclimation, high temperatures increase body core temperature, sweating, metabolic rate, and alter the rate at which glucose and lipids are metabolized. These effects appear to be transitory, as the animal's metabolism returns to near temperate metabolism after 2-3 weeks of acclimatization. High temperatures also result in reduced feed intake. Feeding animals under high temperature conditions generally means keeping forage intake to a minimum and maximizing the energy density of the concentrate. A high fat concentrate is recommended because fat reduces the heat load on the animal, even though the fat may be metabolically less important than glucose. Increased sweat production will increase nitrogen loss, making additional protein intake necessary. Nevertheless, protein intake, above the animal's requirement, may have an adverse effect on body heat production. Therefore, minimizing protein intake, by the addition of appropriate amino acids, such as lysine and threonine, to the diet, may be beneficial.     

In vitro evaluation of cryopreserved bovine sperm and its relation to field fertility in fixed‐time artificial insemination

This study aimed to assess characteristics of bovine cryopreserved sperm and evaluate its relation to field fertility in fixed‐time artificial insemination (FTAI). Semen samples of 16 bulls were used to inseminate 811 Nellore cows, and four of these bulls were also used to inseminate 101 Nellore heifers. Samples of the same ejaculate used for FTAI from each bull were analysed in the laboratory after thawing. Sperm motility and vigour were subjectively assessed by light microscope, and integrity of the plasma and acrosome membranes, and H₂O₂ production were evaluated by flow cytometer. Relation among sperm characteristics and pregnancy rate of cows and heifers were evaluated by univariate and multivariate logistic regression. Subjective sperm motility and vigour did not affect the probability of pregnancy in cows or heifers. In univariate analysis for pregnancy in cows, sperm traits related to acrosome injury positively affected probability of pregnancy mainly when associated with plasma membrane integrity; H₂O₂ production seems to be less important than plasma membrane integrity in affecting probability of pregnancy. In multivariate analysis, sperm traits related to injured acrosome positively affected probability of cow and heifer pregnancies while intact acrosome was negatively related to cow pregnancy. Intact plasma membrane and high H₂O₂ production were positively related to cow pregnancy but negatively related to heifer pregnancy. Results suggest that a capacitation‐like status of the acrosome may benefit probability of pregnancy in cows.     

Changes in caecal microbiota and mucosal morphology of weaned pigs

An experiment was designed to monitor the changes in caecal microbiota associated with early weaning. Twelve piglets (20+/-2 days) from six different litters were selected from a commercial source. For the two experimental groups, one animal from each litter was weaned onto a post-weaning diet (W) and the other remained with the sow (S). After 1 week, animals were sacrificed and caecal samples taken. Microbial counts for total bacteria, enterobacteria and lactobacilli populations were determined by quantitative PCR using SYBR Green dye. Microbial profiles were assessed by terminal restriction fragment length polymorphism (t-RFLP). Weaning promoted an increase in the enterobacteria:lactobacilli ratio (0.27 versus 1.67 log/log 16S rRNA gene copy number, P=0.05). Total bacteria and richness of the caecal microbial ecosystem (number of peaks) were similar in both experimental groups (49.3 for S and 53.4 for W, respectively, P=0.22), although the band patterns were clearly grouped in two different clusters by dendogram analysis. Weaning was also associated with a decrease in crypt density, an increase in mytotic index and a decrease in the number of goblet cells. A reduced immunological response was also observed and was manifested by an increase in intraepithelial lymphocytes and lymphocyte density in the lamina propria. Weaning appears to be critical in the establishment of the caecal microbiota in pigs with important changes, particularly in microbial groups and in caecal mucosal architecture.