Co-aggregation of Lactobacillus and Candida

Co-aggregation study was evaluated as follows: 129AX-1 tubes with slides (Sterilin^TM, Thermo Fisher Scientific Inc., Rodano (MI), Italy) were added with 500 µL of an overnight culture of Lactobacillus strains grown at 37 °C in MRS broth (Oxoid, Thermo Fisher Scientific Inc., Rodano (MI), Italy, CM0359) under micro-aerobic conditions and 500 µL of an overnight culture of Candida spp. grown at 33 °C in DMEM (Dulbecco’s MEM—Biochrom GmbH, Berlin, Germany). Tubes were incubated at 37 °C for 4 h. Then, Gram staining was used to visualize the co-aggregates under oil immersion microscopy (1000×) [30 (link)]. Scoring was done according to Reid et al. [31 (link)]. Moreover, the co-aggregation were also evaluated on Hep-2 and Caco-2 cells. Tubes with monolayer cells were incubated at 37 °C for 4 h after inoculation with Candida and Lactobacillus strains, then Giemsa staining was used to visualize co-aggregates and adhesion on cells.