Agency Response Letter GRAS Notice No. GRN 000445

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CFSAN/Office of Food Additive Safety

April 10, 2013

Robert H. Sindt
Attorney at Law
Suite 110G
1025 Thomas Jefferson Street, NW
Washington, DC 20007

Re: GRAS Notice No. GRN 000445

Dear Mr. Sindt:

The Food and Drug Administration (FDA) is responding to the notice, dated September 21, 2012, that you submitted on behalf of Danisco USA, Inc. (Danisco) 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 September 25, 2012, filed it on October 12, 2012, and designated it as GRAS Notice No. GRN 000445.

The subjects of the notice are Bifidobacterium animalis subsp. lactis strains HN019, Bi-07, B1-04, and B420 (B. animalis subsp. lactis strains). The notice informs FDA of the view of Danisco USA, Inc. that B. animalis subsp. lactis strains are GRAS, through scientific procedures, for use as ingredients in ready-to-eat breakfast cereals, bars, cheeses, milk drinks and milk products, bottled water and teas, fruit juices, fruit nectars, fruit ‘ades’ and fruit drinks, chewing gum, and confections at a maximum level of 2 x 1011 colony forming units (cfu) per serving.

As part of its notice, Danisco includes the report of a panel of individuals (Danisco’s GRAS panel) that evaluated the data and information that are the basis for Danisco’s GRAS determination. Danisco considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Danisco’s GRAS panel discusses the identity and characterization, manufacturing process and specifications, intended conditions of use and exposure levels, published and unpublished safety studies, and the history of use in foods of B. animalis subsp. lactis strains. Based on this review, Danisco’s GRAS panel concluded that B. animalis subsp. lactis strains produced consistent with current good manufacturing practices (cGMP) and meeting appropriate food grade specifications are GRAS under the conditions of their intended use.

Danisco describes the identity of B. animalis subsp. lactis strains, noting that bifidobacteria are Gram-positive, nonsporeforming, anaerobic, pleomorphic bacilli that are predominant among the colonic microflora. Danisco notes that strains Bl-04 and Bi-07 were of human origin, while strains HN019 and B420 were isolated from fermented milk products.

Danisco discusses the manufacture of B. animalis subsp. lactis strains, noting that the company maintains frozen seed vials of each strain and verifies the identity of each strain before fermentation. Each strain is fermented under time- and temperature-controlled conditions. After fermentation, the cultures are pH adjusted and concentrated via centrifugation. Cryoprotectants are added to the concentrated cultures before freezing and freeze-drying. The dried cultures are then packaged and stored under cool and dry conditions. Danisco states that B. animalis subsp. lactis strains are produced in compliance with cGMP. Danisco states that components of the fermentation media are not derived from major food allergens, and the production processes use safe and suitable food grade materials. Danisco reports that freeze-dried samples from production lots of the four strains are stable at 4° C and 25° C for up to 24 months. For each B. animalis subsp. lactis strain, Danisco provides specifications for identity and cell counts, as well as limits for microbial contaminants.

Danisco states that B. animalis subsp. lactis strains are to be used individually in the intended food applications at a maximum level of 2 x 1011 cfu per serving of food. The uses are limited to those foods that can sustain the living organisms for the shelf life of the food. Danisco estimates that consumers will typically ingest a single serving of a food containing one of the B. animalis subsp. lactis strains per day, resulting in exposure to 2 x 1011 cfu per day of the strain. Danisco calculates that higher consumption of 10 servings of these foods per day results in a total consumption of 2 x 1012 cfu of B. animalis subsp. lactis strain(s) per day.

Danisco discusses publicly available safety information about bifidobacteria and B. animalis subsp. lactis. ATCC lists bifidobacteria as Biosafety Level 1 organisms, which the Centers for Disease Control and Prevention defines as organisms not known to cause disease in healthy human adults. B. animalis subsp. lactis is described as nonpathogenic and nontoxigenic in the published literature. Danisco discusses the history of safe use of B. animalis subsp. lactis in food, noting that it is listed in the International Dairy Federation’s “Inventory of Microorganisms with a Documented History of Use in Food” (2002). Further, B. animalis subsp. lactis meets the European Food Safety Authority’s criteria for “Qualified Presumption of Safety” status.

Danisco also describes published information supporting the safe use of strains of B. animalis subsp. lactis. The published information includes animal and human (including infants) studies on HN019, Bi-07, and B1-04. These strains were found to be well-tolerated up to the highest level of ingestion, 2.5 x1012 cfu per kilogram of body weight per day (equivalent to approximately 1.5 x 1014 cfu per day for a 60 kilogram individual). Furthermore, Danisco cites a published in vitro study confirming that strain B420 does not contain virulence factors associated with pathogenicity.

Danisco discusses the potential for transmissible antibiotic resistance from B. animalis subsp. lactis strains. Danisco provides unpublished bioinformatic analyses showing that HN019, Bi-07, B1-04, and B420 each contain a tetracycline-resistance gene (tet(W)) with the same chromosomal structure described in the literature for other B. animalis subsp. lactis strains. Danisco cites a published study that found no evidence of transfer of tet(W) to other bacteria. Based on this information, Danisco concludes that the tet(W) gene found in B. animalis subsp. lactis strains poses minimal risk for transmission of antibiotic resistance to other bacteria. Danisco also describes unpublished in vitro studies showing that B. animalis subsp. lactis strains are sensitive to clinically relevant antibiotics other than tetracycline.

Based on the totality of the data and information described above, Danisco concludes that B. animalis subsp. lactis strains are GRAS for the intended uses in food.

Standards of Identity

In the notice, Danisco states its intention to use B. animalis subsp. lactis strains in several food categories, including foods for which standards of identity exist, located in Title 21 of the Code of Federal Regulations. We note that an ingredient that is lawfully added to food products may be used in a standardized food only if it is permitted by the applicable standard of identity.

Potential Labeling Issues

In describing the human studies conducted using B. animalis subsp. lactis strains, Danisco includes studies that mention potential health benefits. These studies raise a potential issue under the labeling provisions of the Federal Food, Drug, and Cosmetic Act (FD&C Act). Under section 403(a) of the FD&C Act, a food is misbranded if its labeling is false or misleading in any particular. Section 403(r) of the FD&C Act lays out the statutory framework for the use of labeling claims that characterize the level of a nutrient in a food or that characterize the relationship of a nutrient to a disease or health-related condition. If products that contain B. animalis subsp. lactis strains bear any claims on the label or in labeling, such claims are the purview of the Office of Nutrition, Labeling, and Dietary Supplements (ONLDS) in the Center for Food Safety and Applied Nutrition. The Office of Food Additive Safety neither consulted with ONLDS on this labeling issue nor evaluated the information in your notice to determine whether it would support any claims made about B. animalis subsp. lactis strains on the label or in labeling.

Section 301(ll) of the FD&C Act

The Food and Drug Administration Amendments Act of 2007, which was signed into law on September 27, 2007, amends the FD&C Act to, among other things, add section 301(ll). Section 301(ll) of the FD&C Act prohibits the introduction or delivery for introduction into interstate commerce of any food that contains a drug approved under section 505 of the FD&C Act, a biological product licensed under section 351 of the Public Health Service Act, or a drug or a biological product for which substantial clinical investigations have been instituted and their existence made public, unless one of the exemptions in section 301(ll)(1)-(4) applies. In its review of Danisco’s notice that B. animalis subsp. lactis strains are GRAS for the intended uses, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing B. animalis subsp. lactis strains. Accordingly, this response should not be construed to be a statement that foods that contain B. animalis subsp. lactis strains, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).


Based on the information provided by Danisco, as well as other information available to FDA, the agency has no questions at this time regarding Danisco’s conclusion that B. animalis subsp. lactis strains are GRAS under the intended conditions of use. The agency has not, however, made its own determination regarding the GRAS status of the subject use of B. animalis subsp. lactis strains. As always, it is the continuing responsibility of Danisco 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 responding to GRN 000445, as well as a copy of the information in this notice that conforms to the information in the GRAS exemption claim (proposed 21 CFR 170.36(c)(1)), is available for public review and copying at


Dennis M. Keefe, Ph.D.
Office of Food Additive Safety
Center for Food Safety and Applied Nutrition

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