Enumeration and survival studies of free and encapsulated Lactobacillus Acidophilus and Bifidobacterium Lactis in Cheddar cheese

The regulatory standards set by food authorities globally for probiotic foods such as Cheddar cheese makes it essential to have reliable enumeration media that will accurately monitor the survival of probiotic bacteria over the shelf life of Cheddar cheese. This study therefore investigated various selective and differential media for reliable enumeration of Lactobacillus acidophilus, Bifidobacterium spp., starter lactic acid bacteria (SLAB) and non-starter lactic acid bacteria (NSALB) from Cheddar cheese using pure cultures and commercial probiotic Cheddar cheese. All media showed variation in counts and selectivity. Some reported selective media failed to inhibit SLAB and NSLAB. The media that were reliable and also gave good recovery were, Reinforced Clostridium Agar with Bromocresol green and Clindamycin (RCABC), which was selective for L. acidophilus spp. and Reinforced Clostridium Agar with Aniline blue and Dicloxacillin (RCAAD), which was differential for Bifidobacterium spp. and SLAB. Reinforced Clostridium Agar with Bromocresol green and Vancomycin (RCABV) was found suitable for NSLAB. Additionally, an enzyme based colorimetric assay was modified successfully and used as a confirmatory test to check the presence of bifidobacterial colonies on enumeration media. Six batches of probiotic Cheddar cheese were manufactured with the incorporation of LAFTI L10 (L. acidophilus) and LAFTI B94 (B. lactis). The survival of probiotic bacteria, SLAB and NSLAB were monitored over a six-month ripening period using the selected media. The survival of free probiotic bacteria throughout the ripening process decreased consistently in all the six batches. In order to enhance the survival of probiotic bacteria, the effect of microencapsulation on the viability of LAFTI L10 and LAFTI B94 in Cheddar cheese was studied. Six batches of Cheddar cheese were manufactured with the incorporation of alginate-starch encapsulated and free cells of LAFTI L10 and LAFTI B94. The survival of both the encapsulated and free probiotic bacteria was studied over a six month ripening period. The survival of encapsulated LAFTI L10 and LAFTI B94 (107 cfu/g) was found to be significantly better than that of free bacteria (105 cfu/g) at the end of six months of ripening period in Cheddar cheese