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To evaluate the relationship between douching and bacterial vaginosis (BV) among women with and without prior abnormal vaginal flora.1193 women had vaginal swabs obtained for Gram stain for BV, culture for vaginal microflora, and DNA amplification for Neisseria gonorrhoeae and Chlamydia trachomatis at baseline and 6, 12, 24, and 36 months. Self-reported douching was queried every 6 months.In cross-sectional analyses, douching at least once per month was associated with BV among women who had BV at the immediately preceding visit but not among women with normal or intermediate flora. In prospective analyses, douching increased the risk of acquisition for BV among women with intermediate flora (adj. HR 1.5, 95% CI 1.1–2.4) but not among women with normal flora.Douching appeared to be associated with BV among women with already imbalanced flora but not among women with normal flora.