An aliquot of the extracted CW was used to isolate autoch- thonous LAB microflora from the coconut. A 200 ml aliquot of CW was incubated at 37 °C for 48 h in aerobic conditions. In order to isolate colony-forming units (CFU) of cultivable popu- lations, 1 ml of naturally fermented CW was serially diluted tenfold in 0.05 mole/l sodium citrate (Sigma) buffer, pH 7.5, and
100 l of each dilution were spread on plates of MRS (de Man Rogosa Sharpe) agar (Acumedia, Baltimore, MD, USA) pH 5.4 in triplicate and incubated at 37 °C for 48 h as described by De Dea Lindner et al. (2008). More than 100 CFUs were isolated and
purified in MRS broth (Acumedia). To identify LAB within the isolated strains, preliminary tests including microscopic mor- phology characteristics, Gram staining, catalase production, CO2 production from glucose and organic acid (lactic and acetic) production were performed. Only seven wild coconut LAB strains were detected and subsequently tested for viability after a freeze challenge. These strains were cultured onto MRS broth and stored for 48 h at −80 °C.