Probiotic bacteria survive in Cheddar cheese and modify populations of other lactic acid bacteria. 2014

B Ganesan, and B C Weimer, and J Pinzon, and N Dao Kong, and G Rompato, and C Brothersen, and D J McMahon
Dairy Technology and Innovation Laboratory, Western Dairy Center, Utah State University, Logan, UT, USA; Department of Nutrition, Dietetics, and Food Sciences, Utah State University, Logan, UT, USA.

OBJECTIVE Starter lactic acid bacteria in Cheddar cheese face physico-chemical stresses during manufacture and ageing that alter their abilities to survive and to interact with other bacterial populations. Nonstarter bacteria are derived from milk handling, cheese equipment and human contact during manufacture. Probiotic bacteria are added to foods for human health benefits that also encounter physiological stresses and microbial competition that may mitigate their survival during ageing. We added probiotic Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei and Bifidobacterium animalis subsp. lactis to full-fat, reduced-fat and low-fat Cheddar cheeses, aiming to study their survival over 270 days of ageing and to determine the role of the cheese matrix in their survival. RESULTS Probiotic and other lactic acid bacterial populations were enumerated by quantitative PCR using primers specifically targeting the different bacterial genera or species of interest. Bifidobacteria were initially added at 10(6) CFU g(-1) cheese and survived variably in the different cheeses over the 270-day ageing process. Probiotic lactobacilli that were added at 10(7) CFU g(-1) cheese and incident nonstarter lactobacilli (initially at 10(8) CFU g(-1) cheese) increased by 10- to 100-fold over 270 days. Viable bacterial populations were differentiated using propidium monoazide followed by species-specific qPCR assays, which demonstrated that the starter and probiotic microbes survived over ageing, independent of cheese type. Addition of probiotic bacteria, at levels 100-fold below that of starter bacteria, modified starter and nonstarter bacterial levels. CONCLUSIONS We demonstrated that starter lactococci, nonstarter lactobacilli and probiotic bacteria are capable of surviving throughout the cheesemaking and ageing process, indicating that delivery via hard cheeses is possible. Probiotic addition at lower levels may also alter starter and nonstarter bacterial survival. CONCLUSIONS We applied qPCR to study multispecies survival and viability and distinctly enumerated bacterial species in commercial-scale Cheddar cheese manufacture.

UI MeSH Term Description Entries
D007778 Lactobacillus A genus of gram-positive, microaerophilic, rod-shaped bacteria occurring widely in nature. Its species are also part of the many normal flora of the mouth, intestinal tract, and vagina of many mammals, including humans. Lactobacillus species are homofermentative and ferment a broad spectrum of carbohydrates often host-adapted but do not ferment PENTOSES. Most members were previously assigned to the Lactobacillus delbrueckii group. Pathogenicity from this genus is rare.
D008892 Milk The off-white liquid secreted by the mammary glands of humans and other mammals. It contains proteins, sugar, lipids, vitamins, and minerals. Cow Milk,Cow's Milk,Milk, Cow,Milk, Cow's
D002611 Cheese A nutritious food consisting primarily of the curd or the semisolid substance formed when milk coagulates. Cheeses
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001644 Bifidobacterium A rod-shaped, gram-positive, non-acid-fast, non-spore-forming, non-motile bacterium that is a genus of the family Bifidobacteriaceae, order Bifidobacteriales, class ACTINOBACTERIA. It inhabits the intestines and feces of humans as well as the human vagina.
D015169 Colony Count, Microbial Enumeration by direct count of viable, isolated bacterial, archaeal, or fungal CELLS or SPORES capable of growth on solid CULTURE MEDIA. The method is used routinely by environmental microbiologists for quantifying organisms in AIR; FOOD; and WATER; by clinicians for measuring patients' microbial load; and in antimicrobial drug testing. Agar Dilution Count,Colony-Forming Units Assay, Microbial,Fungal Count,Pour Plate Count,Spore Count,Spread Plate Count,Streak Plate Count,Colony Forming Units Assay, Microbial,Colony Forming Units Assays, Microbial,Agar Dilution Counts,Colony Counts, Microbial,Count, Agar Dilution,Count, Fungal,Count, Microbial Colony,Count, Pour Plate,Count, Spore,Count, Spread Plate,Count, Streak Plate,Counts, Agar Dilution,Counts, Fungal,Counts, Microbial Colony,Counts, Pour Plate,Counts, Spore,Counts, Spread Plate,Counts, Streak Plate,Dilution Count, Agar,Dilution Counts, Agar,Fungal Counts,Microbial Colony Count,Microbial Colony Counts,Pour Plate Counts,Spore Counts,Spread Plate Counts,Streak Plate Counts
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D050296 Microbial Viability Ability of a microbe to survive under given conditions. This can also be related to a colony's ability to replicate. Bacterial Viability,Virus Viability,Bacteria Viability,Microbial Inactivation,Inactivation, Microbial,Viability, Bacteria,Viability, Bacterial,Viability, Microbial,Viability, Virus
D019344 Lactic Acid A normal intermediate in the fermentation (oxidation, metabolism) of sugar. The concentrated form is used internally to prevent gastrointestinal fermentation. (From Stedman, 26th ed) Lactate,2-Hydroxypropanoic Acid,2-Hydroxypropionic Acid,Ammonium Lactate,D-Lactic Acid,L-Lactic Acid,Propanoic Acid, 2-Hydroxy-, (2R)-,Propanoic Acid, 2-Hydroxy-, (2S)-,Sarcolactic Acid,2 Hydroxypropanoic Acid,2 Hydroxypropionic Acid,D Lactic Acid,L Lactic Acid,Lactate, Ammonium
D019936 Probiotics Live microbial DIETARY SUPPLEMENTS which beneficially affect the host animal by improving its intestinal microbial balance. Antibiotics and other related compounds are not included in this definition. In humans, lactobacilli are commonly used as probiotics, either as single species or in mixed culture with other bacteria. Other genera that have been used are bifidobacteria and streptococci. (J. Nutr. 1995;125:1401-12) Probiotic

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