INTRODUCTION: Risky sexual behavior has been explored in women living with HIV (WLHIV) internationally but is not well studied in WLHIV in the United States (U.S.). This merits further investigation due to the negative reproductive and HIV health outcomes associated with risky sexual behavior, such as the increased risk for HIV transmission and infertility from sexually transmitted infections (STIs). The aims of this study are to (1) describe sexual behaviors in a cohort of WLHIV in Florida, (2) determine whether demographic characteristics, substance use, and mental health symptoms are associated with risky sexual behavior in a cohort of WLHIV in Florida, and (3) explore whether the relationship between substance use and mental health symptoms and risky sexual behavior differs in reproductive-age (age 18-49), compared to non-reproductive-age WLHIV (age 50+). DESIGN: This was a cross-sectional analysis of data from a multisite cohort study done in Florida. METHODS: Data were collected from a sample of 304 participants who were recruited into the Florida Cohort Study from 9 clinical and community sites in Florida between 2014 to 2017. The predictor variables of interest were mental health symptoms, substance use, and demographic variables. The outcome variable of interest was risky sexual behavior which was defined as reporting ≥1 of the following: (1) at least one STI diagnosis in the past 12 months, (2) two or more sexual partners in the past 12 months, or (3) any inconsistent condom use in the past 12 months. Descriptive statistics, bivariate analysis, and logistic regression (p < 0.1) were conducted on the variables of interest. RESULTS: The mean age of the sample was 47.8 years, and approximately half (51.6%) of the sample was of reproductive-age. Reporting ≥1 risky sexual behavior was reported in over half (51.6%) of the reproductive-age WLHIV in the sample and 32% of the non-reproductive-age WLHIV in the sample. Binge drinking, alcohol-related problems, marijuana use, and age were all significantly associated with self-reporting ≥1 risky sexual behaviors in all WLHIV. Self-reporting binge drinking, marijuana use, and a high alcohol-related problems score respectively were associated with increased odds of self-reporting ≥1 risky sexual behavior in all WLHIV. Neither mental health symptoms nor race/ethnicity or education were significantly associated with self-reporting ≥1 risky sexual behavior in all WLHIV. Self-reporting severe anxiety symptoms and high alcohol-related problems scores respectively were associated with increased odds of self-reporting ≥1 risky sexual behavior only in reproductive-age WLHIV from the sample. CONCLUSION: Marijuana use, binge drinking, and alcohol-related problems appear to have a relationship with risky sexual behavior in WLHIV regardless of age. Reproductive-age also appears to influence risky sexual behavior in WLHIV, and specific reporting severe anxiety symptoms and high alcohol-related problems in reproductive-age WLHIV increases the odds of engaging in risky sexual behavior. CLINICAL SIGNIFICANCE: This study holds clinical significance for nurses and other clinicians working in reproductive health settings and clinics with WLHIV. Results indicate that it could be beneficial to do more screening for mental health symptoms (particularly anxiety) and alcohol use in younger reproductive-age WLHIV.