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Abstract

In individual dogs, despite good quality of raw sperm, some parameters are significantly changed after thawing, which cannot be predicted. We therefore investigated whether motility parameters objectively obtained by CASA, membrane integrity (MI), cell morphology or a combination are suitable to improve the prediction of bad post-thaw quality. For this purpose 250 sperm analysis protocols from 141 healthy stud dogs, all patients introduced for sperm cryopreservation, were evaluated and a Classification and Regression Tree (CART) -analysis performed. The sperm was routinely collected, analysed, and frozen by using a modified Uppsala system. After thawing, data were routinely examined by using CASA, fluorescent microscopy for membrane integrity (MI) and Hancock’s fixation for evaluation of cell morphology. Samples were sorted by post-thaw progressive motility (P) in good (P > / = 50%, n=135) and bad freezers (P<50%, n=115). Among bad freezers, 73.9% showed in addition post-thaw total morphological abberations of >40% and/or MI <50%. Bad freezers were significantly older than good freezers (p<0.05). Progressive motility (P), velocity curvilinear (VCL), mean coefficient (STR), and linear coefficient (LIN) were potential predictors for post-thaw sperm quality since specifity was best (85.8%) and sensitivity (75.4 %) and accuracy (80.4 %) good. For these objectively measured raw sperm parameters, cut-off values were calculated allowing prediction of bad post-thaw results with high accuracy: P = 83.1 % VCL = 161.3 µm/sec, STR = 0.83 %, and LIN = 0.48 %. Raw sperm samples with values below these cut off values will have below average post-thaw quality with a probability of 85.8%. We conclude that VCL, P, STR and LIN are potential predictors of the outcome of sperm cryopreservation, when combined.
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Abstract

A negative energy balance is a common condition in high yielding dairy cows causing the production of ketone bodies (KB), including beta-hydroxybutyrate (BHB), defined as subclinical ketosis (SCK) if clinical signs are missing. The aim of the present study was to evaluate a handheld electronic device for the detection of SCK (BHB-concentration > 1.2 mmol/l), in capillary blood and venous whole blood in cows (WellionVet BELUA, MED TRUST Handels GmbH, Marz, Austria) as well as the feasibility of the puncture of the external vulva with a single use lancet. For this purpose, the blood BHB-concentration was tested in 250 venous and capillary blood samples and compared to the results of a certified laboratory. The majority (76.3%) of the animals displayed no signs of discomfort related to the puncture and in 74.2% the procedure was successful on the first attempt. The BHB-concentrations detected in capillary blood showed good agreement with the reference method, both in capillary (correlation coefficient 0.94 (p<0.001), Kappa-value 0.89) and venous whole blood (correlation coefficient of 0.95 (p<0.001), Kappa-value 0.89). Altogether, 98% of all the samples were correctly classified as SCK or non-SCK by the handheld device in capillary blood (sensitivity 0.96, specificity 0.98) and 97.4% in venous whole blood (sensitivity 0.889, specificity 0.991), respectively. An increase in the correlation by the adaptation of the cut off level could not be achieved for both sampling sites.
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