Effects of autochthonous microbial community on the die-off of fecal indicators in tropical beach sand.

The recently observed high levels of fecal indicators in beach sand confound beach water monitoring efforts. The high levels of fecal indicators may be caused by the loss or the reduced activities of common environmental stresses controlling die-off in the sand. Microcosm experiments were conducted to compare the effects of biotic stresses from autochthonous sand bacteria, protozoa, and viruses on Escherichia coli and Enterococcus faecalis in two tropical beach sands. The inhibition of protozoan activities by cycloheximide did not significantly affect the die-off of E. coli, indicating that protozoan predation played a limited role in beach sand. The contribution from phage infection to E. coli die-off was also negligible. Consequently, autochthonous bacteria were identified as the predominant biotic stress to the die-off of E. coli in beach sand. Subsequent experiments demonstrated that the beach sand had a very low protozoan concentration and low protozoan growth potential when compared with various environmental samples. Co-culturing of E. coli with autochthonous sand bacterial isolates significantly enhanced E. coli die-off. PCR-denaturing gradient gel electrophoresis analysis revealed a complex sand bacterial community, suggesting that bacterial antagonistic effects may be widespread. The study also found that E. faecalis exhibited a much longer survival in beach sand compared with E. coli.