Aeromonas hydrophila, a gastrointestinal pathogen of humans, was shown to exhibit a significant adaptive acid tolerance response (ATR) capable of protecting cells from severe acid at a pH of 3.5. The ATR was induced by exposure to a relatively mild pH level of 5.0 for 20 min. Adaptation required protein synthesis since treatment with chloramphenicol during adaptation to pH 5.0 prevented the development of acid tolerance. The adaptation to acid environment was found to be a non-transient phenomenon. Also, iron was not required for acid adaptation in A. hydrophila. Two-dimensional protein analyses revealed an increased production of 28 proteins and decreased synthesis of 10 following pH shifts from 7.2 to 5.0. The mild pH treatment must act as a signal to A. hydrophila to adapt and survive in acid environments by producing 'protective' proteins. The adaptation and survival of this pathogen in low pH may provide valuable information about its ability to withstand acid environments in nature and in the human gastrointestinal tract.