Table 1.
Enumeration of cell types of 10 Clostridium botulinum strains by phase contrast microscopy. The table shows the average and standard deviation of the percentage of cells belonging to three replicates except for UN1/10-7B strain (only two replicates).
| Group | Strain | Hours | Non-Sporulating Cells | Sporulating Cells | Free Spores | Free Phase-Dark Spores | Total Cell Number |
|---|---|---|---|---|---|---|---|
| I | ATCC 3502 | 5 | 100 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 1009 |
| 24 | 96 ± 2.0 | 4.0 ± 2.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 1108 | ||
| 48 | 84 ± 7.1 | 16 ± 7.1 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 2484 | ||
| 72 | 79 ± 5.7 | 19 ± 5.5 | 1.4 ± 0.6 | 0.0 ± 0.0 | n = 2829 | ||
| ATCC 19397 | 5 | 100 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 798 | |
| 24 | 98 ± 0.3 | 1.2 ± 0.3 | 0.1 ± 0.2 | 0.0 ± 0.0 | n = 1127 | ||
| 48 | 95 ± 1.4 | 4.0 ± 1.2 | 0.4 ± 0.2 | 0.0 ± 0.0 | n = 1044 | ||
| 72 | 90 ± 0.7 | 6.6 ± 1.2 | 2.9 ± 0.8 | 0.0 ± 0.0 | n = 1975 | ||
| UN1/10-7B | 5 | 100 | 0.0 | 0.0 | 0.0 | n = 692 | |
| 24 | 98 | 1.8 | 0.0 | 0.0 | n = 853 | ||
| 48 | 64 | 35 | 0.4 | 0.0 | n = 716 | ||
| 72 | 50 | 48 | 0.2 | 0.0 | n = 757 | ||
| ATCC 17841 | 5 | 100 ± 0.2 | 0.0 ± 0.0 | 0.2 ± 0.2 | 0.0 ± 0.0 | n = 1636 | |
| 24 | 98 ± 1.6 | 2.1 ± 1.5 | 0.1 ± 0.1 | 0.0 ± 0.0 | n = 1281 | ||
| 48 | 75 ± 11 | 25 ± 12 | 0.2 ± 0.2 | 0.0 ± 0.0 | n = 1721 | ||
| 72 | 54 ± 21 | 42 ± 21 | 2.9 ± 1.7 | 0.3 ± 0.2 | n = 1116 | ||
| UN5/11-8 | 5 | 100 ± 0.1 | 0.0 ± 0.1 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 1553 | |
| 24 | 99 ± 0.4 | 1.0 ± 0.4 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 3510 | ||
| 48 | 88 ± 2.4 | 11 ± 2.2 | 1.3 ± 0.3 | 0.0 ± 0.0 | n = 1083 | ||
| 72 | 90 ± 1.3 | 6.8 ± 1.6 | 3.4 ± 0.4 | 0.0 ± 0.0 | n = 1219 | ||
| II | Eklund 17B | 8 | Low cell density, no pellet was obtained | ||||
| 24 | 100 ± 0.2 | 0.1 ± 0.2 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 2166 | ||
| 48 | 75 ± 4.2 | 6.8 ± 1.8 | 18 ± 5.9 | 0.1 ± 0.2 | n = 1036 | ||
| 72 | 76 ± 4.3 | 0.3 ± 0.1 | 22 ± 4.1 | 0.9 ± 0.8 | n = 1208 | ||
| Beluga | 8 | 98 ± 1.3 | 2.2 ± 1.3 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 3390 | |
| 24 | 80 ± 3.8 | 4.8 ± 1.4 | 15 ± 4.2 | 0.0 ± 0.0 | n = 831 | ||
| 48 | 38 ± 1.1 | 5.6 ± 2.2 | 56 ± 1.9 | 0.0 ± 0.0 | n = 370 | ||
| 72 | 7.7 ± 4.5 | 0.5 ± 0.9 | 75 ± 4.6 | 17 ± 7.4 | n = 550 | ||
| CB11/1-1 | 8 | Low cell density, no pellet was obtained | |||||
| 24 | 100 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 1346 | ||
| 48 | 85 ± 1.0 | 3.8 ± 1.4 | 9.0 ± 1.5 | 2.5 ± 0.6 | n = 1138 | ||
| 72 | 60 ± 18 | 1.9 ± 0.9 | 23 ± 12 | 15 ± 7.8 | n = 1130 | ||
| III | Stockholm C | 8 | 100 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | n= § |
| 24 | 94 ± 0.5 | 3.1 ± 1.4 | 3.0 ± 1.4 | 0.0 ± 0.0 | n = 829 | ||
| 48 | 82 ± 2.6 | 7.4 ± 1.5 | 8.9 ± 1.2 | 1.5 ± 1.0 | n = 788 | ||
| 72 | 60 ± 2.2 | 11 ± 4.4 | 27 ± 3.1 | 1.2 ± 1.1 | n = 457 | ||
| BKT015925 | 8 | 100 ± 0.1 | 0.2 ± 0.1 | 0.0 ± 0.0 | 0.0 ± 0.0 | n = 2603 | |
| 24 | 94 ± 1.3 | 1.2 ± 0.8 | 4.9 ± 1.2 | 0.0 ± 0.0 | n = 899 | ||
| 48 | 65 ± 21 | 1.7 ± 0.7 | 33 ± 21 | 0.0 ± 0.0 | n = 302 | ||
| 72 | 46 ± 5.0 | 1.1 ± 2.0 | 45 ± 6.4 | 7.5 ± 6.2 | n = 427 | ||
§ In early time points, Stockholm C formed extremely long cells that divided into normal-sized cells. It proved to be difficult to determine which cells were individual cells and which cells were still part of an elongated cell. However, no cells showed signs of sporulation.