Reactive oxygen species (ROS) production from spermatozoa has been mea
sured by chemiluminescence in the two fractions of a Percoll gradient
column (47 and 90%), Chemiluminescent signals were recorded in each fr
action after the addition of luminol and horse-radish peroxidase (basa
l state), and after stimulation with formyl-methionyl-leucyl-phenylala
nine and phorbol ester (PMA), Oligozoospermic samples show a higher ra
te of ROS production than the normozoospermic samples in both fraction
s of Percoll. Also, ROS were generated at a higher rate by asthenozoos
permic samples in the 90% Percoll fraction than by normal samples afte
r stimulation with PMA, Our data confirm the fact that white blood cel
ls play a major role in the production of ROS, even after purification
on a Percoll gradient, Immunological cases were also found to be asso
ciated with an increased production of ROS, which may be caused by the
same underlying pathological condition responsible for the production
of the antibodies. Repeated centrifugation of the samples triggers a
burst of ROS in excess of that produced after Percoll preparation. In
addition, superoxide dismutase activity was found to be significantly
increased in cases with an elevated production of ROS, It is concluded
that measuring the ROS generation by semen may yield useful informati
on on the functional capacity of spermatozoa, which may be used to imp
rove the success of male infertility management.