Effects of dibromochloropropane and ethylene dibromide on biochemical events and ultramorphology of ejaculated ram spermatozoa in vitro

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Abstract
The effects of dibromochloropropane (DBCP) and ethylene dibromide (EDB) on the function and ultrastructure of freshly ejaculated washed ram spermatozoa were assessed. These two compounds inhibited the collective motility of the sperm in a dose‐dependent fashion when the sperm cells generated their energy either by mitochondrial respiration (2‐deoxyglucose‐treated sperm cells) or by the intact energy production system. DBCP and EDB inhibited the oxygen uptake by the sperm in a dose‐dependent manner. No change in lactic acid accumulation and glucose utilization by the sperm cells was noted following DBCP and EDB addition. No change in the collective motility of the sperm was noted when DBCP or EDB were added to spermatozoa treated with the electron transfer inhibitor, antimycin A. Electron microscopy studies of sperm cells treated with DBCP revealed lesions in the plasma membrane adjacent to the acrosome and in the acrosomal membrane forming vesiculations. The inner membrane and the matrix space of the mitochondria were condensed following DBCP treatment, leaving a large mitochondrial peripheral space, compared with the control. EDB, at the concentration studied, caused no change in the ultramorphological structure of the sperm. DBCP was more potent, at least 4‐fold, compared with EDB. An in vitro direct effect of DBCP and EDB on ram spermatozoa was established. It is suggested that quantitative measurements of sperm collective motility derived by different metabolic pathways can be used as an in vitro toxicological model for evaluation of toxicological and environmental factors affecting biological systems.