Quantification of eumelanin and pheomelanin: stereologic image analysis method

Chemical antioxidants and catalase have been shown to be ineffective in improving the motility of ram spermatozoa in a chemically-defined diluent (RSD-1). In an attempt to identify the biochemical basis of this observation, the activity of aromatic amino acid oxidase (AAAO), the enzyme responsible for generation of hydrogen peroxide in ram and bull spermatozoa, has been investigated. Ram spermatozoa contained higher levels of AAAO activity than bull spermatozoa, although the physico-chemical properties of the enzyme were generally similar in both species. Components of the medium had a marked effect upon AAAO activity. In the presence of glutamate and 3-(N-morpholino)propanesulfonic acid (MOPS), AAAO activity was decreased. Pyruvate appeared to increase AAAO activity. This was due to the ability of this substance to destroy hydrogen peroxide. Pyruvate in RSD-1 works as an effective antioxidant and therefore eliminates the need for other antioxidants in the semen diluent.