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3. safety evaluation > (4) Ames Test

Ames Test of earthplusTM with bacteria
[Test Reference: B010756]
Consigner: Shinshu Ceramics Co., Ltd.
Tested by: Japan Oil Assessment Association Foundation Total Analysis Center
Experiment Results and Observation
The experiment results are as shown in table 1 and 2, and diagram 1 through 5.
In this experiment, weather with or without S9mix, the number of reversely mutated colonies induced by the test substance was less than the number of such colonies in experimental control.
This result is repeatable because the same results were acquired in dose-finding test as well.
Contamination of saprophyte was not found in sterilization test.
No inhibition of bacteria growth was shown in the strain growth test.
Deposition was found on plate, so a cross “+” was drawn to show the amount of the deposition.

Total viable bacteria count in the used strain was as following:
Strain Base pair substitution Frameshift
TA100 TA1535 WP2uvrA- TA98 TA1537
Tvb.count Dose-FindingTest 3.7 3.7 5.5 3.9 2.0
(X 109/ml) This experiment 3.8 4.3 4.7 3.9 2.6

Whether if it was with or without S9mix, the number of reversely mutated colonies induced by the test substance in positive control exceeded twice the number of such colonies in the experimental control, the result was positive,
and the average values of experimental control and positive control were within the range defined in the background data as provided by Analysis Institute , which proves that the test was executed properly.
Based on the results in this experiment, earthplusTM does not cause reverse mutation.