Differentiation of ejaculates showing reactive oxygen species production by spermatozoa or leukocytes

Differences between subgroups and correlations between reactive oxygen species (ROS), sperm motility, concentration of leukocytes and viability in semen samples from 143 men were investigated. Patients with azoospermia or leukocytospermia were excluded from the study. Spermatozoa were separated by means of glass wool filtration. Reactive oxygen species were determined by means of luminol chemiluminescence before and after sperm separation; thereafter, normozoospermic and oligozoospermic patients were divided into three subgroups using the mean of all patients investigated (17462 count 10(-7) viable spermatozoa) as cut-off value as follows: G1--high reactive oxygen species production in native semen and after glass wool filtration; G2--high production of reactive oxygen species only in native semen; G3--low levels of reactive oxygen species before and after glass wool filtration. In general, reactive oxygen species were significantly higher in oligozoospermic samples than in normozoospermic samples. In men with normal sperm count, ROS production correlated significantly with the number of leukocytes in the ejaculate before glass wool filtration, but not thereafter. Glass wool filtration is useful to distinguish between sperm samples in which reactive oxygen species are generated by leukocytes and those in which reactive oxygen species are excessively generated by the spermatozoa themselves.