Survival of Lactobacillus leichmannii in Relation to Vitamin B 12 Assays

Since washed cells ofLactobacillus leichmannii ATCC 7830 frequently showed erratic results in vitamin B12 assays, a depletion technique was used to stabilize the inoculum. The method consisted of incubating the twice-washed cells at 37 C for 45 min in distilled water. Survival and growth studies indicated that the stabilizing procedure (i) did not affect cell numbers, (ii) was optimal for depleting possible carry-over of vitamin B12 or other nutrilites and reserves, and (iii) brought about a phasing of the cells of the inoculum which underwent logarithmic growth immediately on inoculation. This minimal period (45 min) was the same when the cells were incubated in dextrose-water. Survival of the cells in physiological saline was greater than in distilled water, but a longer time was required for stabilizing the inoculum in saline, which precluded its use in routine work. The prepared inoculum showed an improvement over the washed inoculum in that readings between duplicate and triplicate tubes agreed closely and the standard curves were reproducible whether acidimetric or turbidimetric methods were followed. No erratic results were noted in more than 1 year of testing.