TABLE 1.
Bacterial reverse mutation test with pasteurized A. muciniphila
| Concentration (μg/plate) | Revertant colonies per plate (mean ± standard deviation) | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Without metabolic activation (–S9) | With metabolic activation (+S9) | |||||||||
| S. typhimurium | S. typhimurium | |||||||||
| TA98 | TA100 | TA102 | TA1535 | TA1537 | TA98 | TA100 | TA102 | TA1535 | TA1537 | |
| Plate incorporation assay (experiment 1) | ||||||||||
| 0 (water) | 14.3 ± 2.5 | 130.7 ± 15.6 | 455.7 ± 31.8 | 9.0 ± 3.6 | 7.7 ± 1.5 | 24.3 ± 5.5 | 114.3 ± 9.1 | 520.3 ± 56.1 | 11.0 ± 4.6 | 8.3 ± 3.8 |
| 2.29 | – | – | – | – | – | 20.0 ± 2.6 | – | – | – | – |
| 6.86 | – | – | – | – | – | 31.0 ± 5.0 | – | – | – | – |
| 20.6 | 16.3 ± 3.8 | 129.3 ± 11.7 | 501.0 ± 39.0 | 17.0 ± 0.0 | 4.0 ± 1.0 | 30.3 ± 8.1 | 116.0 ± 23.6 | 482.3 ± 34.8 | 11.0 ± 4.6 | 6.7 ± 2.3 |
| 61.7 | 17.3 ± 4.6 | 131.0 ± 14.7 | 396.3 ± 162.3 | 12.7 ± 3.2 | 6.3 ± 1.5 | 18.7 ± 4.5 | 115.3 ± 17.0 | 558.3 ± 60.7 | 12.3 ± 4.2 | 6.7 ± 2.3 |
| 185.2 | 13.0 ± 1.0 | 128.3 ± 17.2 | 354.0 ± 27.0 | 17.3 ± 2.1 | 10.0 ± 1.0 | 22.7 ± 2.3 | 117.7 ± 11.7 | 530.0 ± 25.4 | 10.3 ± 2.1 | 7.7 ± 3.5 |
| 555.6 | 19.0 ± 5.6 | 131.7 ± 11.1 | 479.3 ± 73.9 | 13.0 ± 5.0 | 9.3 ± 0.6 | 24.7 ± 4.0 | 132.3 ± 3.1 | 468.3 ± 60.3 | 13.3 ± 1.2 | 7.7 ± 2.3 |
| 1666.7 | 18.3 ± 4.0 | 119.0 ± 10.0 | 386.7 ± 46.3 | 14.7 ± 0.6 | 5.3 ± 4.0 | – | 118.0 ± 12.1 | 513.3 ± 28.9 | 12.3 ± 5.0 | 12.0 ± 7.2 |
| 5000 | 20.7 ± 2.1 | 169.0 ± 4.4 | 397.0 ± 75.4 | 17.7 ± 5.0 | 8.7 ± 0.6 | – | 153.0 ± 24.2 | 500.7 ± 60.0 | 16.0 ± 4.6 | 9.7 ± 3.5 |
| Positive control a | 109.3 ± 7.6 | 693.3 ± 27.3 | 1215.3 ± 77.9 | 607.3 ± 60.0 | 83.0 ± 11.3 | 1071.3 ± 30.7 | 913.3 ± 27.1 | 1060.7 ± 30.2 | 113.0 ± 25.5 | 93.0 ± 3.6 |
| Plate incorporation assay (experiment 2) | ||||||||||
| 0 (water) | 18.7 ± 4.5 | 101.0 ± 8.7 | 458.3 ± 34.6 | 6.0 ± 2.6 | 6.3 ± 1.2 | – | – | – | – | – |
| 61.7 | 18.0 ± 3.5 | 95.0 ± 6.0 | 444.3 ± 24.3 | 7.0 ± 2.6 | 8.7 ± 3.1 | – | ||||
| 185.2 | 18.3 ± 9.5 | 122.0 ± 15.4 | 439.0 ± 36.5 | 12.3 ± 3.2 | 6.3 ± 4.5 | – | – | – | – | – |
| 555.6 | 17.7 ± 2.9 | 117.7 ± 25.0 | 510.0 ± 38.0 | 12.3 ± 1.2 | 9.7 ± 2.5 | – | – | – | – | – |
| 1666.7 | 15.7 ± 2.9 | 142.0 ± 25.1 | 453.0 ± 20.9 | 13.7 ± 6.4 | 7.3 ± 3.1 | – | – | – | – | – |
| 5000 | 41.3 ± 4.7 | 136.7 ± 7.5 | 543.0 ± 80.9 | 11.3 ± 2.3 | 10.0 ± 1.7 | – | – | – | – | – |
| Positive control b | 95.7 ± 11.7 | 623.0 ± 34.4 | 1245.7 ± 78.8 | 677.3 ± 28.7 | 160.0 ± 60.9 | – | – | – | – | – |
| Plate incorporation assay (experiment 3) | ||||||||||
| 0 (water) | 14.3 ± 5.1 | – | – | – | – | – | – | – | – | – |
| 312.5 | 15.0 ± 6.2 | |||||||||
| 625 | 17.0 ± 5.0 | – | – | – | – | – | – | – | – | – |
| 1250 | 17.7 ± 2.5 | – | – | – | – | – | – | – | – | – |
| 2500 | 15.7 ± 0.6 | – | – | – | – | – | – | – | – | – |
| 3750 | 13.7 ± 3.2 | – | – | – | – | – | – | – | – | – |
| 5000 | 19.7 ± 8.7 | – | – | – | – | – | – | – | – | – |
| Positive control c | 79.3 ± 9.0 | – | – | – | – | – | – | – | – | – |
| Preincubation assay | ||||||||||
| 0 (water) | – | – | – | – | – | 24.3 ± 4.5 | 153.3 ± 25.0 | 413.7 ± 12.3 | 9.7 ± 0.6 | 5.7 ± 1.5 |
| 2.29 | – | – | – | – | – | – | – | – | – | – |
| 6.86 | – | – | – | – | – | 29.0 ± 3.5 | – | – | – | – |
| 20.6 | – | – | – | – | – | 23.3 ± 4.2 | – | – | – | – |
| 61.7 | – | – | – | – | – | 35.0 ± 2.0 | 161.0 ± 21.6 | 492.3 ± 23.6 | 9.0 ± 2.6 | 5.7 ± 2.1 |
| 185.2 | – | – | – | – | – | 28.7 ± 9.3 | 172.3 ± 13.3 | 451.7 ± 56.1 | 12.0 ± 5.2 | 12.7 ± 5.7 |
| 555.6 | – | – | – | – | – | 26.3 ± 2.3 | 181.7 ± 6.0 | 421.7 ± 22.3 | 9.3 ± 4.5 | 14.3 ± 4.2 |
| 1666.7 | – | – | – | – | – | 24.7 ± 5.1 | 173.3 ± 20.1 | 505.3 ± 36.2 | 13.7 ± 0.6 | 7.7 ± 2.1 |
| 5000 | – | – | – | – | – | – | 199.3 ± 11.0 | 446.3 ± 75.8 | 14.7 ± 5.0 | 12.7 ± 6.0 |
| Positive control d | – | – | – | – | 1453.7 ± 43.7 | 615.3 ± 38.6 | 918.7 ± 45.5 | 149.7 ± 10.4 | 75.7 ± 4.2 | |
–S9, without metabolic activation; +S9, with metabolic activation; 2‐AM, 2‐anthramine; 9‐AA, 9‐aminoacridine, 2‐NF, 2‐nitrofluorene; 4NQO, 4‐nitroquinoline‐1‐oxide; B[a]P, benzo[a]pyrene; MMC: mitomycin C; NaN3, sodium azide; S. typhimurium, Salmonella typhimurium.
Positive controls: –S9: TA98 = 0.5 μg/plate 2‐NF; TA100 and TA1535 = 1 μg/plate NaN3; TA1537 = 50 μg/plate 9‐AA; TA102 = 0.5 μg/plate MMC; +S9: TA98, TA1535 and TA1537 = 2 μg/plate 2‐AM; TA102 = 20 μg/plate 2‐AM; TA100 = 5 μg/plate B[a]P.
Positive controls –S9: TA98 = 0.5 μg/plate 2‐NF; TA100 and TA1535 = 1 μg/plate NaN3; TA1537 = 50 μg/plate 9‐AA; TA102 = 0.5 μg/plate MMC.
Positive controls –S9: TA98 = 0.5 μg/plate 2‐NF.
Positive controls +S9: TA98, TA1535 and TA1537 = 2 μg/plate 2‐AM; TA102 = 20 μg/plate 2‐AM; TA100 = 5 μg/plate B[a]P.