The aim of this pilot study was to establish a reactive oxygen species (ROS) evaluation method as a step in the routine diagnosis of men from infertile couples, which attend the Centre of Assisted Reproduction at the Teaching Hospital in Olomouc. Standard semen analyses were performed manually according to WHO guidelines. The number of peroxidase-positive leukocytes in the semen was determined using the Endtz test. The levels of ROS were estimated by chemiluminescence assay using luminol (5-amino-2,3 dihydro-1,4 phthalazinedione) as a probe. The semen samples were collected from 68 patients. Normospermia was found in 15 patients (22.1 %). The semen samples of 3 normospermic patients were classified as ROS-positive. Elevated ROS production was recorded in all subgroups of patients irrespective of any pathology found. We confirmed that spermatozoa might be the source of ROS as well as the seminal leukocytes. Apart from the leukocytes, sperm cells with residual cytoplasm and immature spermatozoa are considered to be a major source of ROS. Thus it is suggested that sperm morphology abnormalities should be evaluated more carefully.

Institute of Biology Faculty of Medicine Hnevotínská 3 Palacký University 775 15 Olomouc Czech Republic

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