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Background: Hygiene-based practices of lubrication, genital cleansing, postcoital urination, and rectal douching are common behaviors among populations at higher risk of human immunodeficiency virus (HIV)/sexually transmitted infections (STI). Yet, the role these behaviors have on HIV/STI risk has not been well elucidated, especially among transgender women (TW) and gay, bisexual, and other men who have sex with men (GBMSM). Additionally, advances in biomedical strategies have heralded a new era of HIV/AIDS prevention that may be accompanied by behavioral changes that lead to decreases in condom usage and subsequent changes to STI sequelae. Nevertheless, many people at higher risk are not benefiting equally from these options, strengthening the need for more sustainable, evidence-based methods., Objectives: This study explored the knowledge, attitudes, and behaviors of hygiene-based practices, proven preventative methods, and preferred learning methods among Peruvian TW and GBMSM., Methods: 30 interviews and 50 questionnaires were conducted with TW (N=35), GBMSM (N=35), healthcare providers (N=5), and key community informants (N=5)., Results: Most participants perceived hygiene-based practices to be common behaviors and a significant aspect of sexual wellbeing. Educational materials utilizing social media and hosting in-person events were also viewed favorably, with value to enhancing HIV/STI knowledge., Conclusions: Several barriers to autonomy surfaced in the data, including systemic disparities to adequate HIV/STI services, top vs. bottom social dynamics, and PrEP accessibility issues. Continued work is needed to address the barriers to the acceptability, feasibility, and potential efficacy of hygiene-based practices, biomedical/barrier strategies, and L 3 +., Competing Interests: COMPETING INTERESTS The authors have no competing interests.