Hello. I want to ask if anyone knows methods for survivability in simulated GIT fluids and also aggregation (auto- and co-aggregation) using well plates (24 or 96 wells) for lactic acid bacteria? We want to use an absorbance microplate reader to hold the experiments.
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Screening and characterization of lactic acid bacterial strains that produce fermented milk and reduce cholesterol levels Authors: Xuefang Guan, Qingxian Xu, Yi Zheng, Lei Qian, Bin Lin ∗
Cultures of HLX37 and Escherichia coli DH5 were grown in MRS and Luria-Bertani medium (LB, Canspec, Inc., China) overnight, respectively, and then centrifuged at 3000 × g for 10 min at 4 ◦C. The cells were washed twice, resuspended in phosphate-buffered solution (PBS), and adjusted to an initial OD600 of 0.4 ± 0.1 (A0). The cell suspensions were then incubated at room temperature at 25 ◦C for 24 h to allow phase separation, and the final OD600 value (A24) of the aqueous phase was measured. The percentage of auto-aggregation was expressed as follows: Aauto% = (A0 - A24)/A0 × 100.15 Similarly, equal volumes of LAB and E. coli DH5 were mixed, adjusted to an initial OD600 of 0.6 ± 0.1 (A0) in PBS and incubated at room temperature for 24 h without agitation. The OD600 value (A24) of the aqueous phase was measured. The co-aggregation percentage was expressed as follows: Aco% = (A0 - A24)/A0 × 100.
I have used the protocol in this paper in the past to determine auto and co-aggregation. It worked well. but was for a small study (not published).
Article Safety Evaluation and Colonisation Abilities of Four Lactic ...
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