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2 protocols using blood agar plates

1

Listeria monocytogenes Strain Cultivation

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D-(+)-glucose, sodium polyacrylate, acetic acid 99–100% (glacial), nitric acid (≥65%), hydrogen peroxide 30% (w/w), Triton X-100, sodium tartrate dehydrate, sodium hydrogen carbonate, ascorbic acid and phosphate buffered saline powder were obtained from Sigma-Aldrich (Steinheim, Germany). Copper (II) sulfate-pentahydrate and ethanol were purchased from Merck (Darmstadt, Germany); bicinchoninc acid disodium salt and sodium carbonate decahydrate from Alfa Aesar (Haverhill, MA, USA); and 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) from Enzo Life Science (Farmingdale, NY, USA). The olive oil (a mixture of refined and virgin olive oils) was purchased in a supermarket in Italy. Ultrapure water was obtained from a Milli-Q Plus system (Millipore, Bedford, MA, USA) and was always used freshly prepared.
Brain heart infusion broth (BHI), listeria selective agarb (LSM), and blood agar plates were produced by Biolife, while plate count agar (PCA) was produced by Biokar diagnostics. The reference strain for experimental trials was the American Type Culture Collection (ATCC 19117), which came from a Listeria monocytogenes (serotype 4d) third line culture to prevent the risk of phenotypic alteration.
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2

Culturing Oral Pathogenic Bacteria

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Reference strains Streptococcus mutans ATCC 25175, Streptococcus oralis ATCC 6249, Streptococcus salivarius K12 ATCC BAA-1024, and Aggregatibacter actinomycetemcomitans ATCC 29522 (Microbiologics, St Cloud, MN, USA) were used. Bacteria were grown on blood agar plates (Biolife, Milan, Italy) supplemented with 5% sheep blood (Biognost, Zagreb, Croatia) in anaerobic conditions at 37 °C for 24–48 h.
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