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Cette seconde culture est incubée une nuit à 30 °C. Enfin, 100 mL de cette seconde culture servent à ensemencer 2 L de lait. Cette dernière culture est incubée à 30 °C pendant 19 h pour atteindre un pH de 4,84 et une acidité de 55 ° Dornic. La culture est stockée une nuit à 4 °C avant utilisation pour la fabrication fromagère ,