Sensitivity of Escherichia coli to Seaweed (Ascophyllum nodosum) Phlorotannins and Terrestrial Tannins

Pure culture experiments were conducted to assess the bacteriostatic and bactericidal effects of phlorotannins (PT) isolated from Ascophyllum nodosum (brown seaweed) on Escherichia coli O157:H7. In Exp. 1, one non-O157:H7 strain (25922) and three strains of E. coli O157:H7 (3081, EDL933 and E318N) were cultured in M9 medium with PT included at 0 (control), 25, 50 or $100{\mu}g/ml$ (n = 3). Bacterial growth was monitored by $OD_{600}$ at 0, 4, 6, 12 and 24 h, and by dilution plating at 0, 4, 6 and 24 h. All strains were inhibited (p<0.001) by PT to varying degrees. At 50 or $100{\mu}g/ml$, PT prevented growth of all four strains. At $25{\mu}g\;PT/ml$, growth of 25922, 3081, E318N and EDL933 was inhibited for 6, 12 and 24 h, respectively, but 25922 and 3081 resumed growth by 12 and 24 h. Direct plating confirmed bactericidal effects of PT on all four strains at $100{\mu}g/ml$, and on EDL933 and E318N at $50{\mu}g/ml$. In Exp. 2, strains 25922 and 3081 were incubated with no tannins or with $50{\mu}g/ml$ of PT, purified condensed tannins (CT) from Quebracho (Schinopsis balansaei), or purified tannic acid from Rhus semialata (Anacardiaceae) as hydrolysable tannins (HT). Strain 3081 was unaffected by HT or CT, but was completely inhibited (p<0.001) by PT at 4, 6 and 24 h. Strain 25922 was unaffected by HT, slightly inhibited by CT, and almost eradicated by PT at 4 and 6 h. Transmission electron microscopy revealed tannin-mediated alterations to bacterial cell walls. Phlorotannins from A. nodosum exhibit growth-inhibiting and bactericidal effects in vitro against the strains of E. coli O157:H7 investigated. Anti-E. coli efficacy of A. nodosum PT is superior to that of terrestrial tannins purified from Quebracho and from Rhus semialata.