In both batch and continuous culture, Peptostreptococcus anaerobius was able to grow in vaginal defined medium with Prevotella bivia, but not in pure culture. Growth of P. anaerobius was increased by 238% ( P<0.001) in peptone-supplemented vaginal defined medium conditioned by prior growth of P. bivia. Analysis of P. bivia culture supernatants showed a net accumulation of amino acids and subsequent growth of P. anaerobius in the conditioned supernatants resulted generally in amino acid utilization. Supplementation of peptone-supplemented vaginal defined medium with amino acids in concentrations similar to those available after prior growth with P. bivia were growth-stimulatory (246%, P=0.006) for P. anaerobius. Increased availability of amino acids by P. bivia is proposed as a mechanism to support the observed in vitro commensal symbiosis between P. bivia and P. anaerobius.