direction of Dr. William Flowers).Four experiments were conducted to determine the effects of time; dilution; slide location; and technician on common sperm quality estimates recorded using a computer-assisted sperm analysis (CASA) system. For each experiment, the split ejaculate technique was used with 18 boars being collected weekly for 5 to 6 weeks (n=100 to 130 observations per treatment per experiment). For analysis time, CASA measurements were obtained at 0, 30, 60, 90 and 120 s after loading the sample onto the evaluation chamber. Motility, progressive motility, curvilinear velocity (VCL), straight line velocity (VSL) and cell beat frequency (BCF) all decreased (p<0.05) over time with their highest measurements recorded at either 0 or 30 s. Amplitude of lateral head displacement (ALH) remained constant (p=0.23). For dilution, the neat sample was analyzed along with subsamples diluted with BTS extender in ratios of 1:1, 1:2, and 1:3.Increasing the dilution of neat semen had variable effects on CASA parameters. However, measurements for the 1:1 rate were consistently equal to or higher (p<0.05) than those recorded for neat semen and their counterparts with higher dilutions. For slide location, CASA measurements were recorded from 5 locations on the slide: lower central; middle central; upper central; middle right; and upper right. The location within the chamber at which measurements were taken had no effect on VCL (p=0.32); VSL (p=0.45); and BCF (p=0.14). For motility, progressive motility, and ALH all locations were similar (p>0.05) with the exception of the middle central in which the lowest (p<0.05) readings were recorded. For the effect of technician, 5 individuals with different levels of expertise with microscopes loaded and recorded CASA measurements from the same slide according to a randomized schedule. No differences (p>0.67) among technicians were observed for any of the CASA variables. Collectively, results from these studies indicate that neat samples should be diluted 1:1 and readings should be taken from any location except middle central slide area within 30 s of loading the sample in order to obtain the highest measurements. Prior experience of the technician performing CASA does not appear to have a significant effect on mobility estimates
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