The gene expression profiles of Escherichia coli strains grown anaerobically with or without Acacia mearnsii (black wattle) extract were compared to identify tannin resistance strategies. The cell envelope stress protein gene spy and the multidrug transporter-encoding operon mdtABCD, both under the control of the BaeSR two-component regulatory system, were significantly up-regulated in the presence of tannins. BaeSR mutants were more tannin sensitive than their wild-type counterparts.Condensed tannins, or proanthocyanidins, are common secondary metabolites of plants, consisting of flavonol polymers (17). High concentrations of tannins in fodder plants inhibit gastrointestinal bacteria (6,9,10,20) and reduce animal performance (9,18,21,23). Numerous mechanisms of how tannins may inhibit bacteria have been proposed, including tannin-polymer complexation, tannin-induced membrane disruption, and metal ion chelation (22). Despite the antimicrobial activities of tannins, many tannin-resistant bacteria have been isolated. However, the mechanisms behind this resistance are still unknown (22). Gram-negative bacteria may be less sensitive to the inhibitory effects of tannins, as gastrointestinal bacterial populations shifted toward gram-negative Enterobacteriaceae and Bacteroides species in rats fed diets containing 0.7 to 2.0% condensed tannins (20). There was a corresponding decrease in the gram-positive Clostridium leptum group. Escherichia coli is resistant to concentrations of up to 1% Acacia mearnsii (black wattle) tannin extract (WTE) under anoxic conditions. However, it was demonstrated to be sensitive to 0.1% under oxic conditions due to auto-oxidation of tannins resulting in hydrogen peroxide generation (19). To get insight into the mechanisms responsible for resistance to condensed tannins under anoxic conditions, we conducted a gene expression study to measure the total transcriptional responses of E. coli in the presence of WTE.The E. coli strains used in this study are described in Table 1. All growth experiments were performed with MOPS (3-morpholinopropane-1-sulfonic acid) medium (12), with glucose (0.4%) as the carbon source, and iron and trace elements were provided by trace element solution SL-10 (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany). MOPS medium was prepared, gassed with N 2 , and dispensed in an anaerobic chamber (Coy Laboratory Products, Ann Arbor, MI) under an atmosphere of 95% N 2 and 5% H 2 . Cysteinesulfide solution as a reducing agent was added to anaerobic medium (the final concentrations of L-cysteine ⅐ HCl and Na 2 S ⅐ 9H 2 O were 0.025%). WTE, an aqueous extract from Acacia mearnsii (black wattle) bark, was added to the MOPS medium at a final concentration of 1% (wt/vol). WTE contained 65.6% phenols as tannic acid equivalents and was donated by Wickett and Craig of America, Inc. (Curwensville, PA). WTE was filter sterilized before addition to growth medium. In order to identify specific transcriptional responses to the presence of tannins, as op...