U. S. Food and Drug Administration
Center for Food Safety and Applied Nutrition
Office of Food Additive Safety
March 19, 2002

Fred Degnan
King and Spalding
1730 Pennsylvania Avenue, N.W.
Washington, DC 20006-4706

Re: GRAS Notice No. GRN 000049

Dear Mr. Degnan:

The Food and Drug Administration (FDA) is responding to the notice, dated June 9, 2000, that you submitted on behalf of Nestlé USA (Nestlé) in accordance with the agency’s proposed regulation, proposed 21 CFR 170.36 (62 FR 18938; April 17, 1997; Substances Generally Recognized as Safe (GRAS); the GRAS proposal). FDA received the notice on June 14, 2000, and designated it as GRAS Notice No. GRN 000049.

The subjects of the notice are Bifidobacterium lactis strain Bb12 and Streptococcus thermophilus strain Th4.⁽¹⁾ This notice informs FDA of the view of Nestlé that B. lactis strain Bb12 and S. thermophilus strain Th4 are GRAS, through scientific procedures, for use as ingredients in milk-based infant formula that is intended for consumption by infants four months and older, at levels not to exceed good manufacturing practice.⁽²⁾ In its GRAS Exemption Claim, Nestlé notes that the intended use of S. thermophilus strain Th4 and B. lactis strain Bb12 does not encompass use by immunocompromised infants.

As part of its notice, Nestlé includes the report of a panel of individuals (Nestlé’s GRAS Panel) who evaluated the data and information that are the basis for Nestlé’s GRAS determination. Nestlé considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food.

Nestlé describes published information about the identity and characteristic properties of B. lactis strain Bb12 derives from a human isolate. In the past, Bifidobacterium was classified within the genus Lactobacillus;⁽³⁾ more recently, analyses of 16S ribosomal RNA gene sequences have been used to establish Bifidobacterium as a distinct genus. Bifidobacteria are Gram-positive, non-motile and non-spore-forming rods indigenous to the intestinal tract of infants, adults, and various animals. Bergey’s Manual of Systemic Bacteriology describes bifidobacteria as anaerobic microorganisms (i.e., colonies of bifidobacteria do not grow on culture plates maintained under aerobic conditions) (Ref. 2). Different species and strains of bifidobacteria exhibit different sensitivity to oxygen.⁽⁴⁾ While strain Bb12 was originally classified at the species level as Bifidobacterium bifidum, it is now classified as B. lactis.

Nestlé describes published information about the identity and characteristic properties of S. thermophilus strain Th4. S. thermophilus strain Th4 derives from a mozzarella cheese isolate. The genus Streptococcus is characterized by Gram-positive spherical or ovoid bacteria that are typically arranged in chains or pairs. Streptococci are non-sporing, catalase-negative, and homofermentative. S. thermophilus strain Th4 is a lactic acid bacterium that can survive and grow in the presence of oxygen, but exclusively generates ATP by fermentation, which is an anaerobic process (Ref. 3).⁽⁵⁾ S. thermophilus is not associated with any pathogenic traits.

Given the public health implications of antibiotic resistant strains of bacteria in the food supply, Nestlé provides information about the antibiotic resistance of B. lactis strain Bb12 and S. thermophilus strain Th4. Nestlé states that B. lactis strain Bb12 is susceptible to various antibiotics. Nestlé states that strains of S. thermophilus that have been tested for antibiotic resistance usually are sensitive to a broad range of antibiotics and that Nestlé does not expect S. thermophilus strain Th4 to present an adverse public health risk.

Nestlé provides information about the method of manufacture for B. lactis strain Bb12 and S. thermophilus strain Th4, including release criteria. During the manufacturing process, Nestlé assesses strain quality, genetic stability, carbohydrate fermentation profile, enzyme profile, and the number of organisms. Nestlé provides release criteria for strain quality. Nestlé also describes its good manufacturing practice/quality assurance program to validate the identity of the organisms in each production lot. This program includes DNA fingerprinting as well as traditional biochemical and morphological techniques.

Nestlé considers that a key purpose for adding these ingredients to infant formula is to help improve the overall wholesomeness of food for infants and children. Nestlé expects that infant formula containing these microbial ingredients will help maintain, in formula-fed infants, an intestinal flora composition resembling that in infants fed human breast milk and that this, in turn will help digestive function.

Nestlé provides information about the level of microorganisms per gram of powdered product and per eating occasion. Nestlé intends to add freeze-dried B. lactis strain Bb12 to the infant formula to give counts of 10⁷-10⁸ colony forming units per gram (cfu/g) final powdered product. Nestlé intends to add freeze-dried S. thermophilus strain Th4 to the infant formula to give counts of 10⁷-10⁸ cfu/g final product. In addition, Nestlé intends to specify that the level of resuscitated organisms would be within the range of 10⁶ to 10⁸ cfu of each organism per gram of finished formula as the target for each batch.

Nestlé describes its marketing experience, in Europe and Asia, with infant formula containing B. lactis strain Bb12, S. thermophilus, or with infant formula containing related ingredients. Nestlé notes that these formulas meet the infant formula recommendations, guidelines or requirements of various organizations.

Nestlé notes that foods containing lactic acid bacteria are broadly consumed throughout the world, and provides information about the prior use of Bifidobacterium spp. or S. thermophilus in food. Bifidobacteria spp. are found frequently in various fermented foods worldwide. The most common food use of S. thermophilus is in yogurt. Nestlé’s GRAS panel reviewed and discussed the practice in Europe of feeding yogurt to very young children. Nestlé reports that several members of its GRAS panel also mentioned that the practice of feeding yogurt to very young children is becoming increasingly common in the United States.

Nestlé describes two clinical studies in which infants consumed B. lactis strain Bb12 and S. thermophilus strain Th4 in a formula. Study endpoints for the first study included diarrhea and rotavirus shedding. Study endpoints for the second study included anthropometrics, product usage, morbidity, and stooling profile. Nestlé notes that no adverse effects were reported for either study

Nestlé also summarizes more than 50 clinical studies in which more than 2,500 infants (including term and preterm infants) or children consumed infant formula or other foods that contained microbial ingredients. These ingredients derived from more than a dozen different cultures or strains (including Bifidobacterium spp. or S. thermophilus). Typical study endpoints included diarrheal episodes, fecal enzymes, fecal flora composition, stool characteristics, gastrointestinal disturbances, feeding tolerance, systemic immune responses and traditional anthropometrics. None of the investigators reported any adverse events during these studies. Infant subjects presented normal growth. Stool characteristics and stooling patterns of infants who consumed infant formula containing microbial ingredients were similar to those of breast fed infants.

In Nestlé’s view, the fact that these microorganisms may be safely used in formula is further supported by data from over 100 adult clinical trials in which lactic acid bacteria were orally administered. These bacteria were used either as single or mixed cultures, ranging from 10⁷ to 10¹¹ cfu/dose, live or heat-killed organisms. The studies investigated a total of 11 Lactobacillus species, 2 Streptococcus species, 5 Bifidobacterium species and 3 groups of unidentified “lactic acid bacteria cultures.” All of the investigated strains were well-tolerated. In the 40 years of study, using data from over 100 adult clinical studies, not one single adverse event was reported by any of more than 8000 subjects.

Based on the information provided by Nestlé, as well as other information available to FDA, the agency has no questions at this time regarding Nestlé’s conclusion that B. lactis strain Bb12 and S. thermophilus strain Th4 are GRAS for their intended use as ingredients in milk-based infant formula that is intended for consumption by infants four months and older at levels not to exceed good manufacturing practice. The agency has not, however, made its own determination regarding the GRAS status of this use of B. lactis strain Bb12 and S. thermophilus strain Th4. As always, it is the continuing responsibility of Nestlé to ensure that food ingredients that the firm markets are safe, and are otherwise in compliance with all applicable legal and regulatory requirements.

In accordance with proposed 21 CFR 170.36(f), a copy of the text of this letter, as well as a copy of the information in Nestlé’s notice that conforms to the information in proposed 21 CFR 170.36(c)(1), is available for public review and copying on the homepage of the Office of Food Additive Safety (on the Internet at http://www.cfsan.fda.gov/~lrd/foodadd.html).

Intended use in infant formula

Under section 412 of the Federal Food, Drug, and Cosmetic Act (FFDCA), a manufacturer of a new infant formula must make a submission to FDA, providing required assurances about the formula, at least 90 days before the formula is marketed. Nestlé should be aware that FDA’s response to Nestlé’s GRAS notice does not alleviate the responsibility of any infant formula manufacturer who intends to market an infant formula that contains B. lactis strain Bb12 and S. thermophilus strain Th4 to make the submission required by section 412.

For your information, FDA recently contracted with the National Academy of Sciences to address broad issues related to the addition of ingredients to infant formula. We expect to make significant findings of any final report issued under that contract available to the public.

Potential label claims

Under section 403(a) of the FFDCA, a food is misbranded if its labeling is false or misleading in any particular. In describing the intended use of B. lactis strain Bb12 and S. thermophilus strain Th4 and in describing the information that Nestlé relies on to conclude that B. lactis strain Bb12 and S. thermophilus strain Th4 are GRAS under the conditions of their intended use, Nestlé raises a number of issues under the labeling provisions of the FFDCA. These issues are the purview of the Office of Nutritional Products, Labeling, and Dietary Supplements (ONPLDS) in the Center for Food Safety and Applied Nutrition. The Office of Food Additive Safety neither consulted with ONPLDS on these labeling issues nor evaluated the information in Nestlé’s notice to determine whether it would support any claims made about B. lactis strain Bb12 and S. thermophilus strain Th4 on the label or in labeling.

References

1. Stiles, M.E. and W.H. Holzapfel, Int. J. Food Microbiol., 1997; 36(1):1-29.

2. Sneath, P. H. A., N. S. Mair, M. E. Sharpe, and J. G. Holt, eds. Bergey’s Manual of Systemic Bacteriology, vol. 2; pp. 1418-1434. Williams and Wilkins, Baltimore, 1986.

3. Neidhardt, F. C., J. L. Ingraham, and M. Schaechter. Physiology of the bacterial cell: a molecular approach; pp.160-161. Sinauer Associates, Inc., Sunderland, MA, 1990.

4. Holt, J. G., N. R. Krieg, P. H. A. Sneath, J. T. Staley, and S. T. Williams, eds. Bergey’s Manual of Determinative Bacteriology, 9th edition; p.532. Williams and Wilkins, Baltimore, 1994.

⁽¹⁾The notice of June 9, 2000 was for the use of Bifidobacterium, Lactobacillus, and S. thermophilus. By a letter of November 14, 2000, Nestlé narrowed the scope of GRN 000049 to B. lactis strain Bb12 and S. thermophilus strain Th4.

⁽²⁾Nestlé states that it is aware of no reason why these ingredients, if GRAS for use in infant formula, would not be GRAS in other foods for infants and children.

⁽³⁾FDA notes that the “lactic acid bacteria” were a group set aside by Orla-Jensen in the 1920's whose major fermentation product is lactic acid. FDA further notes that Bifidobacteria are considered to be part of the lactic acid bacteria on a “physiological” basis as Bifidobacteria produce lactic acid as a major fermentation product. However, as the rRNA profiles became available, it became more clear by taxonomic, morphological, and genetic criteria, that Bifidobacteria are distinct from the Lactobacilli, Streptococci, Leuconostocs, Lactococci and other related organisms (Ref. 1).

⁽⁴⁾Nestlé uses the phrase “facultative anaerobe” to describe this differential sensitivity to oxygen.

⁽⁵⁾Both the notice and Bergey’s Manual of Determinative Bacteriology, 9th edition (Ref. 4) use the phrase “facultatively anaerobic” to describe S. thermophilus.

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