BACKGROUND: Women who have sex with women (WSW) and women who have sex with women and men (WSWM) are frequently perceived to be at low risk for sexually transmitted infections (STIs), although data show that their STI rates are similar to heterosexual women. Little research has examined sexual behaviors, perceptions of STI risk, and practice of safe sex among African American WSW/WSWM living in the Southern United States, a population of women likely to be at high risk for STIs. METHODS: Focus group discussions were conducted with African American WSW/WSWM living in Birmingham, Alabama, to explore their sexual behaviors with women, perceptions of STI risk from female (and male) sexual partners, and practice of safe sex. Digital audio-recordings were transcribed and analyzed using HyperRESEARCH software. RESULTS: Seven focus groups were conducted between August 2011 and March 2012, with 29 total participants. Women reported a broad range of sexual behaviors with female partners. They were more aware of their risk for STI acquisition from male partners than from female partners and felt that their best options for safe sex in their relationships with women were practicing good hygiene and requiring proof of STI testing results. CONCLUSIONS: African American WSW/WSWM in this study were aware of their STI risk, more so with regard to men, and desired accurate information on safer sex options in their sexual relationships with women. Health care providers can assist these women by helping them apply their existing knowledge of heterosexual STI transmission to their female sexual partnerships. Copyright © 2013 American Sexually Transmitted Diseases.