Field and laboratory evidence shows that deeply buried (90–888 m) fine-grained sediments of the Atlantic Coastal Plain contain viable acetogenic microorganisms, and that these microorganisms actively produce organic acids. Concentrations of formate, acetate, and propionate in pore waters extracted from fine-grained sediments ranged from 50 μM to 5 mM and were much higher than in adjacent pore waters associated with sandy sediments (<2 μM). Laboratory studies showed that asceptically cored fine-grained sediments incubated under a H2 atmosphere produced formate and acetate, and that H14CO3− was converted to 14C-acetate and 14C-formate over time. An enrichment culture of these acetogenic microorganisms was recovered from one long-term incubation that showed the presence of several morphologically distinct gram-positive, rod-shaped bacteria. These microorganisms were capable of growth under autotrophic (H2 + CO2), heterotrophic (syringate), and mixotrophic (H2 + CO2 + syringate) conditions. These results suggest that microbial acetogenesis, rather than abiotic processes, is the most important organic acid-producing mechanism during low-temperature (∼ 30 °C) diagenesis of Atlantic Coastal Plain sediments.