Two heterotrophic, thermophilic, sulfur-reducing archaea were isolated from the Guaymas Basin hydrothermal vent. The fermentation of proteinaceous and carbohydrate substrates was examined at 85-degrees-C for each isolate in the presence and absence of elemental sulfur and in coculture with a thermophilic methanogen. The heterotrophic isolates differed with respect to their requirement for sulfur. Both heterotrophic isolates exhibited a mixed organic acid fermentation from proteinaceous substrates; however, acetate was the sole organic acid produced from carbohydrate fermentation. In coculture fermentations with a thermophilic methanogen, the heterotrophic isolates exhibited enhanced growth and fermentation. Interspecies hydrogen transfer and elemental sulfur-reduction may be important microbial processes in deep-sea hydrothermal vent community metabolism.