S salivarius

The strains used in the study came from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cariogenic strains used were Streptococcus mutans (ATCC 25175), S. mitis (ATCC 49456), S. sanguinis (ATCC 10556), S. sobrinus (ATCC 33478), Lactobacillus paracasei (ATCC 11578), Enterococcus faecalis (ATCC 4082), and S. salivarius (ATCC 25975). For all assays performed the bacteria were incubated in Brain Heart Infusion agar (BHI), added with defibrinated sheep blood (5%) in a microaerophilia incubator for 24 h at 37 °C with 10% CO₂, except for E. faecalis and S. salivarius which were incubated aerobically at 37 °C for 24 h.

Authentic P. elliottii (batch number 21811-09) and P. tropicalis (batch number 20511-09) resin samples were obtained from the Brazilian Association of Resinators (ARESB), located in the city of Avaré – SP, Brazil. DHA was isolated from the P. elliottii resin and was obtained by the method previously described by Leandro et al. (2014) (link) (Supplementary Figures S1, S2).
The following bacteria from the American Type Culture Collection (ATCC) collection were used to conduct the antibacterial assays: S. mutans (ATCC 25175), S. mitis (ATCC 49456), S. sanguinis (ATCC 10556), S. sobrinus (ATCC 33478), S. salivarius (ATCC 25975), Lactobacillus casei (ATCC 11578), and Enterococcus faecalis (ATCC 4082). These microorganisms were maintained in a freezer at −80°C in 20% glycerol solution in the Laboratory of Applied Microbiology Research (LaPeMA) – University of Franca (Unifran).

Anaerobic bacteria T. denticola (ATCC 35405), P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) were purchased from ATCC. T. denticola, P. gingivalis and F. nucleatum were grown as described previously at 37°C under oxygen deprived anaerobic conditions in New Oral Spirochete Medium or in Brain-heart Infusion (BHI) broth supplemented with hemin (5 μg ml^-1) and vitamin K (1 μg ml^-1) [143 , 144 (link)]. Purity of spirochete cultures was confirmed by dark field microscopy prior to use in experiments. Oral commensal bacteria S. gordonii (ATCC 10558), S. salivarius (ATCC 13419) and V. parvula (ATCC 10790) were grown overnight at 37°C under anaerobic conditions in BHI broth then harvested for subsequent assays. Gram staining followed by microscopic evaluation and colony morphology on Mitis Salivarius plates were used to confirm purity of cultures. T.denticola lipoologosaccharide (Td-LOS) was a gift from Daniel Grenier, Universite Laval, Quebec, Canada, P.gingivalis lipopolysaccharide (Pg-LPS) and F.nucleatum lipopolysaccharide (Fn-LPS) were provided by Richard P. Darveau, University of Washington, Seattle and V. parvula lipopolysaccharide (Vp-LPS) was provided by Christopher Fenno, University of Michigan, Ann Arbor, MI.