Stable fluoride-resistant mutants of Streptococcus mutans GS-5 were isolated on Todd-Hewitt agar by a step-wise selection procedure. Resistance is defined here as the ability to form colonies in 48 hr. First-step mutants demonstrated six different levels of resistance, ranging from 400 to 1000 μg/mL sodium fluoride. Second-step mutants demonstrated two levels of resistance, one at 1600 and the highest at 3000 μg/mL sodium fluoride. All mutants (originally isolated at pH 7.2) were tested for fluoride resistance at pH 5 and 6. At these lower pH values, all of the mutants demonstrated resistance to fluoride when compared with the parent strain, but at much-reduced levels.
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