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Does ejaculation decrease testosterone levels?

Masturbation is very unlikely to decrease testosterone levels. Semen is not the source of testosterone so ejaculation will not cause levels to change.

All other things being equal, increased sexual activity - including masturbation - tends to increase testosterone levels. But these levels depend upon many factors including age, time of day, overall level of physical activity, and general health.

Psychological factors can cause changes in testosterone levels: levels go up in men who believe they are going to become more sexually active, levels can be affected by whether a person knows in advance that he will be test for testosterone, and so forth.