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. 1998 Jun;64(6):2192-9.
doi: 10.1128/AEM.64.6.2192-2199.1998.

Development of a probiotic cheddar cheese containing human-derived Lactobacillus paracasei strains

G Gardiner  1 , R P Ross, J K Collins, G Fitzgerald, C Stanton
Affiliations

Development of a probiotic cheddar cheese containing human-derived Lactobacillus paracasei strains

G Gardiner et al. Appl Environ Microbiol. 1998 Jun.

Abstract

Cheddar cheese was manufactured with either Lactobacillus salivarius NFBC 310, NFBC 321, or NFBC 348 or L. paracasei NFBC 338 or NFBC 364 as the dairy starter adjunct. These five strains had previously been isolated from the human small intestine and have been characterized extensively with respect to their probiotic potential. Enumeration of these strains in mature Cheddar cheese, however, was complicated by the presence of high numbers (>10(7) CFU/g of cheese) of nonstarter lactic acid bacteria, principally composed of lactobacilli which proliferate as the cheese ripens. Attempts to differentiate the adjunct lactobacilli from the nonstarter lactobacilli based on bile tolerance and growth temperature were unsuccessful. In contrast, the randomly amplified polymorphic DNA method allowed the generation of discrete DNA fingerprints for each strain which were clearly distinguishable from those generated from the natural flora of the cheeses. Using this approach, it was found that both L. paracasei strains grew and sustained high viability in cheese during ripening, while each of the L. salivarius species declined over the ripening period. These data demonstrate that Cheddar cheese can be an effective vehicle for delivery of some probiotic organisms to the consumer.

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Figures

FIG. 1
FIG. 1
RAPD-PCR-generated DNA fingerprints for probiotic Lactobacillus strains L. salivarius NFBC 310 (lanes 2 and 3), L. salivarius NFBC 321 (lanes 4 and 5), L. paracasei NFBC 338 (lanes 6 and 7), L. salivarius NFBC 348 (lanes 8 and 9), and L. paracasei NFBC 364 (lanes 10 and 11). Lanes 1 and 12 contain 100-bp ladders.
FIG. 2
FIG. 2
(A) Survival of lactobacilli and starter during cheese ripening in trial 1 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to D) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C and D) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lane 19 (B) or 11 (C and D) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 3
FIG. 3
(A) Survival of lactobacilli and starter during cheese ripening in trial 2 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to E) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C to E) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lanes 11 (B to E) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 3
FIG. 3
(A) Survival of lactobacilli and starter during cheese ripening in trial 2 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to E) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C to E) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lanes 11 (B to E) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 3
FIG. 3
(A) Survival of lactobacilli and starter during cheese ripening in trial 2 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to E) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C to E) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lanes 11 (B to E) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 3
FIG. 3
(A) Survival of lactobacilli and starter during cheese ripening in trial 2 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to E) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C to E) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lanes 11 (B to E) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 3
FIG. 3
(A) Survival of lactobacilli and starter during cheese ripening in trial 2 laboratory-scale Cheddar cheeses manufactured under microbiologically controlled conditions. (B to E) RAPD-PCR profiles of a representative number of Lactobacillus isolates from each of the cheeses. Lanes 1 (C to E) show the RAPD profile of each probiotic Lactobacillus strain added to the cheese at manufacture, while a 100-bp ladder is shown in lanes 11 (B to E) and all other lanes show RAPD profiles of Lactobacillus isolates from 6-month (180-day)-ripened cheeses.
FIG. 4
FIG. 4
(A) Survival of lactobacilli and starter during cheese ripening in trial 3 pilot-scale Cheddar cheeses. (B) RAPD-PCR profiles of a representative number of Lactobacillus isolates from vat 2 cheese to which L. paracasei NFBC 364 was added during manufacture. Lane 1 shows the RAPD profile of the added strain. A 100-bp ladder is shown in lane 10, while lanes 2 to 9 and 11 to 20 show RAPD profiles of Lactobacillus isolates from the 6-month (180-day)-ripened cheese.
FIG. 4
FIG. 4
(A) Survival of lactobacilli and starter during cheese ripening in trial 3 pilot-scale Cheddar cheeses. (B) RAPD-PCR profiles of a representative number of Lactobacillus isolates from vat 2 cheese to which L. paracasei NFBC 364 was added during manufacture. Lane 1 shows the RAPD profile of the added strain. A 100-bp ladder is shown in lane 10, while lanes 2 to 9 and 11 to 20 show RAPD profiles of Lactobacillus isolates from the 6-month (180-day)-ripened cheese.
FIG. 5
FIG. 5
Concentration of individual FAA in water-soluble extracts of 6-month-old control and experimental Cheddar cheeses of trial 1 (A) and trial 2 (B).
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