Three methods of preservation of bacteria were studied observing the viability and stability of some morphological and biochemical characteristics of Bacteroides fragilis, Clostridium perfringens, C. difficile, Fusobacterium nucleatum, Peptostreptococcus sp and Prevotella melaninogenica (B. melaninogenicus) strains. After periods of storage, B. fragilis showed the best viability in lyophilization and freezing methods, whereas Clostridium strains were the most resistant bacteria, surviving up to one year after lyophilization, freezing or subculturing. Peptostreptococcus and P. melaninogenica lost the viability at the begining of storage, in all methods studied. The stability of colonial, morphological and physiologic-biochemical aspects of all strains seemed have not been altered throughout this study.