12 well plates

These assays were performed independently co-culturing each one of the H. pylori strains with each one of the different Candida strains, following the protocol described by Sánchez-Alonzo et al. [33 (link)]. Suspensions of each yeast and bacterial strain were adjusted to an O.D. of 0.1 at 600 nm in 0.89% saline solution (SS). Then, 500 µL of H. pylori strain suspension and 500 µL of Candida strain suspension were placed in the well of 12-well plates (Thomas Scientific, Swedesboro, NJ, USA) containing 4 mL BB-5%FBS. The plates were kept at 37 °C for 48 h under the aforementioned conditions. The microaerobic and anaerobic conditions were also achieved using CampyGen or Anaerogen sachets, respectively, as described in the section above. Each co-culture was performed in triplicate. The methodology of this section is shown in Figure S2.

Each H. pylori strain was co-cultured with each Candida spp. strain. A suspension of each strain of each kind of microorganism was independently prepared in 0.89% saline solution (SS) and adjusted to an OD of 0.1 at 600 nm. Then, 500 µL of each suspension of the bacterial strains were placed in wells of 12-well plates (Thomas Scientific, Swedesboro, NJ, USA) previously loaded with 4 mL de BB-5%FBS. Then, 500 µL of yeast cells from each strain were independently added to wells previously inoculated with H. pylori. Four wells were required for each bacterial strain in order to be independently co-cultured with each of the yeast strains. Plates containing the co-cultures were incubated under microaerobic conditions for 48 h at the following temperatures: 4 °C, 20 °C, 37 °C or 40 °C. Microaerobiosis atmosphere was achieved using CampyGen envelopes (Thermo Scientific, Waltham, MA, USA). All assays were repeated thrice. As controls for this assay, pure inocula of all H. pylori and Candida strains were used.

Individual suspensions of Candida and H. pylori strains were prepared in SS at an O.D. of 0.1 at a wavelength of 600 nm. Four mL of BB supplemented with 1%, 5% or 20% FBS or 4 mL of SS was placed in wells of 12-well plates (Thomas Scientific, Swedesboro, NJ, USA), and then 500 µL of each strain was added to each well to obtain the co-cultures. The co-cultures were incubated at 37 °C for 48 h under microaerobic conditions in an incubator (Thermo Scientific, Waltham, MA, USA). Each co-culture was repeated thrice.