We tested the capacity of the AviMate sperm quality analyzer to predict sperm mobility phenotype. A preliminary experiment was performed to determine a sperm concentration that afforded maximal sperm quality index (SQI) values with sperm from males with high sperm mobility. In order to facilitate comparison of sperm motility indexes, semen was diluted to aconstant concentration of 1.25 million sperm/mL rather than a constant ratio of 1:75 as recommended for the sperm quality analyzer. Thereafter, sperm mobility, motile concentration, and SQI were measured from a single ejaculate collected from each of 70 broiler breeders. Coefficients of variation were 60, 37, and 30%, respectively. Neither sperm mobility (r = 0.01) nor motile concentration (r = 0.14) was highly correlated with SQI. Neither motile concentration nor straight line velocity (VSL) differed (P > or = 0.05) among roosters whose SQI values were below or above one standard deviation from the population mean (n = 10 per group). In contrast, both motile concentration (P < or = 0.01) and VSL (P < or = 0.05) differed between roosters whose sperm mobility values were below or above a standard deviation from the population mean (n = 10 per group). Sperm mobility was a function of the size of a highly motile subpopulation of sperm. A replicate experiment was performed with a flock of New Hampshire roosters (n = 126). Neither the properties of motile sperm nor fertility differed among males characterized by extreme SQI scores. We concluded that variation in SQI scores was enigmatic because sperm concentration was controlled in our experiments, sperm viability was invariant, and motile concentration did not differ among roosters characterized by extreme SQI scores. In summary, the AviMate sperm quality analyzer did not predict sperm mobility phenotype.