Bacterial Vesicle Production Protocols

Lactobacillus reuteri DSM 17938 [42 (link)], BioGaia AB, Stockholm, Sweden], and Helicobacter pylori ATCC 43629, were used for this study. The choice of these two microorganisms linked to the fact that in previous works we have demonstrated that they produce vesicles containing DNA, which are released both from the planktonic and the biofilm phenotypes [15 (link),16 (link)]. L. reuteri DSM 17938 was spread on deMan, Rogosa, Sharpe Agar (MRS) (Oxoid Limited, Hampshire, UK), and incubated at 37 °C for 24 h in an anaerobic atmosphere (Anaerogen Pak Jar, Oxoid Ltd., Basingstoke, UK), while H. pylori ATCC43629 was plated on Chocolate Agar (Oxoid Ltd.) supplemented with 1% (v/v) of IsoVitaleX (Becton Dickinson, Franklin Lakes, NewJersey, NJ, USA) and 10% (v/v) of defibrinated horse sterile blood (Oxoid Ltd.), and finally incubated at 37 °C for 3 days in a microaerophilic atmosphere (Campy PakJar; Oxoid Ltd.).