[Federal Register Volume 71, Number 160 (Friday, August 18, 2006)]
[Rules and Regulations]
[Pages 47729-47732]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: E6-13621]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
21 CFR Part 172
[Docket No. 2002F-0316 (formerly 02F-0316)]
Food Additives Permitted for Direct Addition to Food for Human
Consumption; Bacteriophage Preparation
AGENCY: Food and Drug Administration, HHS.
ACTION: Final rule.
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SUMMARY: The Food and Drug Administration (FDA) is amending the food
additive regulations to provide for the safe use of a bacteriophage
preparation on ready-to-eat meat and poultry products as an
antimicrobial agent against Listeria monocytogenes. This action is in
response to a petition filed by Intralytix, Inc.
DATES: This rule is effective August 18, 2006. Submit written or
electronic objections and requests for a hearing by September 18, 2006.
See section VII of this document for information on the filing of
objections. The Director of the Office of the Federal Register approves
the incorporation by reference in accordance with 5 U.S.C. 552(a) and 1
CFR part 51 of certain publications in new 21 CFR 172.785 as of August
18, 2006.
ADDRESSES: You may submit objections and requests for a hearing,
identified by Docket No. 2002F-0316 (formerly 02F-0316), by any of the
following methods:
Electronic Submissions
Submit electronic objections in the following ways:
Federal eRulemaking Portal: http://www.regulations.gov.
Follow the instructions for submitting comments.
Agency Web site: http://www.fda.gov/dockets/ecomments.
Follow the instructions for submitting comments on the agency Web site.
Written Submissions
Submit written objections in the following ways:
FAX: 301-827-6870.
Mail/Hand delivery/Courier [For paper, disk, or CD-ROM
submissions]: Division of Dockets Management (HFA-305), Food and Drug
Administration, 5630 Fishers Lane, rm. 1061, Rockville, MD 20852.
To ensure more timely processing of objections, FDA is no longer
accepting objections submitted to the agency by e-mail. FDA encourages
you to continue to submit electronic objections by using the Federal
eRulemaking Portal or the agency Web site, as described in the
Electronic Submissions portion of this paragraph.
Instructions: All submissions received must include the agency name
and Docket No(s). and Regulatory Information Number (RIN) (if a RIN
number has been assigned) for this rulemaking. All objections received
will be posted without change to http://www.fda.gov/ohrms/dockets/default.htm, including any personal information provided. For detailed
instructions on submitting objections, see the ``Objections'' heading
of the SUPPLEMENTARY INFORMATION section of this document.
Docket: For access to the docket to read background documents or
objections received, go to http://www.fda.gov/ohrms/dockets/default.htm
and insert the docket number, found in brackets in the heading of this
document, into the ``Search'' box and follow the prompts and/or go to
the Division of Dockets Management, 5630 Fishers Lane, rm. 1061,
Rockville, MD 20852.
FOR FURTHER INFORMATION CONTACT: Raphael A. Davy, Center for Food
Safety and Applied Nutrition (HFS-265), Food and Drug Administration,
5100 Paint Branch Pkwy., College Park, MD 20740, 301-436-1272.
SUPPLEMENTARY INFORMATION:
I. Background
In a notice published in the Federal Register of July 22, 2002 (67
FR 47823), FDA announced that a food additive petition (FAP 2A4738) had
been filed by Intralytix, Inc., c/o Lewis & Harrison, 122 C St. NW.,
suite 740, Washington, DC 20001, now represented by Keller & Heckman
LLP, 1001 G St., NW., suite 500 West, Washington, DC 20001. The
petition proposed to amend the food additive regulations to provide for
the safe use of a mixture of bacteriophages\1\ (phages) as an
antimicrobial agent against Listeria monocytogenes (L. monocytogenes)
on foods, including fresh meat, meat products, fresh poultry, and
poultry products. On December 18, 2003, the petitioner amended the
petition to limit the petitioned use to ready-to-eat (RTE) meat and
poultry products only.\2\
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\1\ Bacteriophages are viruses that infect bacteria only.
\2\ Ready-to-eat products, as used in this final rule, are
defined in 9 CFR 430.1.
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The food additive consists of a mixture of equal proportions of six
individually purified phages. The petitioner's rationale for
incorporating multiple phages in one formulation is to minimize the
possibility of L. monocytogenes developing a resistance to the
additive. Each phage in the additive is specific against various L.
monocytogenes strains, including those strains known to be associated
with foodborne illness (e.g., L. monocytogenes strains, serotypes 1/2a,
4b and 1/2b). The phages are lytic\3\ double-stranded DNA phages. The
petitioner has characterized each phage with respect to physical
properties and other appropriate identifying factors (e.g., host range,
structural protein profile, and DNA sequence of complete genome\4\).
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\3\ Lytic bacteriophages lyse (destroy) their host bacteria as a
normal part of their life cycle without integrating into the host
genome.
\4\ Genome means the genetic content of a cell or virus.
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In the manufacturing process, each phage contained in the additive
is separately produced using a strain of L. monocytogenes that can
serve as a host to the specific phage. The host L. monocytogenes strain
is first cultured in microbiological media and the specific phage is
added to the culture when a specified cell density is achieved. After
phage multiplication, which results in lysis (destruction) of host
cells, the phage is purified by use of multiple filtration steps (to
remove bacteria and their components). The six phages produced by this
process are then
[[Page 47730]]
blended in phosphate buffered saline solution to formulate the
additive. The six phages contained in the additive have been deposited
with the American Type Culture Collection\5\ (ATCC).
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\5\ ATCC is a nonprofit bioresource center that maintains
deposits of bacteria and bacteriophages among other biological
materials. Their primary mission is to acquire, authenticate,
preserve, develop, and distribute biological material.
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The phage preparation will be used as an antimicrobial agent to
control L. monocytogenes in the production of RTE meat and poultry
products. The phage preparation is directly sprayed on the surface of
the RTE food articles at a level of approximately 1 milliliter (mL) of
the preparation per 500 square centimeters (cm\2\) of food surface area
just prior to packaging.
II. Determination of Safety
Under the general safety standard in section 409 of the Federal
Food, Drug, and Cosmetic Act (the act) (21 U.S.C. 348), a food additive
cannot be approved for a particular use unless a fair evaluation of the
data available to FDA establishes that the additive is safe for that
use. FDA's food additive regulations (21 CFR 170.3(i)) define safe as
``a reasonable certainty in the minds of competent scientists that the
substance is not harmful under the intended conditions of use.''
In evaluating the safety of the petitioned substance, FDA
considered the following factors in determining the safety of the
proposed food additive use: (1) The safety of the six phages
constituting the food additive; (2) the safety of potential residues
from L. monocytogenes used in the manufacture of the food additive and
the need for limits related to their levels; (3) whether undesirable
genes are potentially carried by the food additive; and (4) the need
for additional identity and safety specifications.
A. Safety of the Petitioned Use of the Phage Preparation
Phages infect only bacteria, rather than mammalian or plant
cells.\6\ Moreover, phages are ubiquitous and humans are routinely
exposed to them at high levels through food, water, and the environment
without adverse effect.\7\ Phages also are a part of the normal
microbial population of the humangut.\8\ However, the petitioner's
bacteriophages are specific to L. monocytogenes only. Therefore, FDA
concludes that the food additive under consideration does not present a
toxicological concern for use in food as proposed by the petitioner
based upon the explanations provided in the following sections (Refs. 1
and 3).
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\6\ T.D. Brock and M.T. Madigan, 1998, Biology of
Microorganisms, 5th edition; Prentice-Hall, Inc., Inglewood Cliffs,
NJ.
\7\ Bergh, O., K.Y. Borsheim, G. Bratbak, and M. Heldal, 1989,
High abundance of viruses found in aquatic environments, Nature,
vol. 340 (10): 467-468.
\8\ Breitbart et al., 2003; Journal of Bacteriology 185 (20):
6220-6223.
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B. Safety Evaluation of Potential Residue Components From L.
monocytogenes
FDA considered the possibility that the proposed food additive may
contain L. monocytogenes components as residues from use of the
organism as host for phage multiplication in the manufacturing process.
Such residues may include the toxin Listeriolysin O (LLO). Potential
residues of L. monocytogenes other than LLO do not present a safety
concern (Ref. 1). Based on our review of scientific literature on the
pathogenicity of L. monocytogenes (Ref. 1), FDA finds that LLO is the
only substance known to be toxic that may potentially be present as a
residue in this food additive after the manufacturing process.
LLO was not detected in the finished food additive within the assay
limits of detection of 5 hemolytic units\9\ (HU)/ml, and the petitioner
provided information on the purification process used in the production
of the food additive as additional assurance that LLO would not be
present at detectable levels in the finished food additive.
Nevertheless, the agency has calculated a worst-case exposure to LLO
from consumption of food products treated with the phage preparation.
Assuming LLO is present at a maximum level of 5 HU/ml in the additive,
the worst-case exposure to LLO for males aged 20 years or more that
consume RTE foods treated with the additive at the maximum intended use
level is 52 HU/person/day (HU/p/d) at the mean and 104 HU/p/d at the
90th percentile. Males aged 20 years or more represent the worst-case
scenario because this population group consumes the highest amount of
food intended to be treated with the additive (Ref. 2). In this safety
evaluation, FDA reviewed all available information on the identity,
toxicity, and the stability of LLO. Even if LLO were present at the
level of 5 HU/ml, this level does not present a toxicological concern
for the following reasons (Ref. 1):
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\9\ 1 HU of LLO is equal to one nanogram of protein (as reported
in Geoffroy, C. et al. 1987, Purification, Characterization, and
Toxicity of the Sulfhydryl-Activated Hemolysin Listeriolysin O from
Listeria monocytogenes, Infection and Immunity, vol. 55(7): pp.
1641-1646).
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1. Inactivation of LLO by Cholesterol
The toxicity of LLO has been shown to be significantly reduced (by
as much as 200- to 2000-fold) following pre-incubation of LLO with
added cholesterol in vitro (Ref. 1). Since the phage preparation will
be used on meat and poultry products and these products normally
contain significant (milligram) amounts of cholesterol, then any
residual amounts of LLO at levels no greater than 5 HU/ml that may be
present in the additive are likely to be inactivated by the
cholesterol.
2. pH and LLO Activity
Studies show that LLO activity is lost or significantly decreased
in acidic (low pH of less than 4) environments (Ref. 1). Residual
amounts of LLO, if present, are likely to be inactivated by the low pH
(less than 4) within the human stomach.
3. Inactivation of Orally Consumed LLO by Human Defense Mechanisms
In vivo studies demonstrate that both normal intestinal microflora
and cell-mediated immunity reactions in the intestines inhibit LLO
(Ref. 1). These defense mechanisms provide some protection against low
incidental oral exposures to LLO (no greater than 5 HU/ml).
Additionally, at these levels, LLO is expected to be rapidly and
irreversibly degraded by proteolytic enzymes that may be presented in
the diet or in the stomach. Thus, LLO at these residual levels would
not pose a toxic threat to humans.
Considering all of the above factors, FDA concludes that potential
residues of LLO that may be found in the food additive are negligible
(5 HU/ml or less) and do not pose a safety concern for the use of the
additive as an antimicrobial agent on RTE meat and poultry products.
Although LLO was not detected in the food additive, the agency
concludes that a specification is necessary to ensure that LLO is not
present in detectable amounts to ensure the purity and safe use of the
petitioned food additive. Thus, the agency is including in this
regulation a specification of not more than 5 HU/ml for LLO (the limit
of detection for the method).
C. Undesirable Genes (Bacterial Toxin Genes) Potentially Carried by
Phages
Lysogenic phages, as opposed to those that are lytic, have the
capacity to integrate into the host genome and may facilitate transfer
of toxin or drug resistance genes between bacterial cells. FDA has
determined that the phages contained in the petitioned food additive
are lytic based on the petitioner's information on host lysis
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characteristics and on genomic analysis of each phage (Ref. 4).
Therefore, FDA concludes that the use of this food additive would not
result in the spread of toxin or drug genes.
D. The Need for Other Specifications
We are also including specifications for potency, absence of
undesirable genes, phage titer\10\, absence of L. monocytogenes and
other microbiological pathogens, and total organic carbon (Ref. 2).
These specifications ensure the identity and safe use of the additive.
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\10\ A term that refers to the number of phage particles per
milliliter of phage solution.
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III. Other Considerations
FDA recognizes that while this rule is issued under the authority
of the Federal Food, Drug, and Cosmetic Act, use of the ingredient must
also comply with the Federal Meat Inspection Act or the Poultry
Products Inspection Act, which are administered by the U.S. Department
of Agriculture (USDA). In particular, those statutes provide that the
ingredient must be suitable for its intended use. FDA recognizes that
there may be meat or poultry products considered RTE for which use of
the additive may not be suitable within the meaning of those statutes.
This regulation addresses only the safety standard under section 409 of
the Federal Food, Drug, and Cosmetic Act and does not address
requirements for suitability administered by the USDA.
IV. Conclusion
FDA reviewed data in the petition and other available relevant
material to evaluate the safety of the use of a phage preparation as an
antimicrobial agent against L. monocytogenes on RTE meat and poultry
products. Based on this information, the agency concludes that the
proposed use of the additive is safe. Therefore, the regulations in
part 172 (21 CFR part 172) should be amended as set forth in this
document.
In accordance with Sec. 171.1(h) (21 CFR 171.1(h)), the petition
and the documents that FDA considered and relied upon in reaching its
decision to approve the petition are available for inspection at the
Center for Food Safety and Applied Nutrition by appointment with the
information contact person (see FOR FURTHER INFORMATION CONTACT). As
provided in Sec. 171.1(h), the agency will delete from the documents
any materials that are not available for public disclosure before
making the documents available for inspection.
V. Environmental Impact
The agency has previously considered the environmental effects of
this rule as announced in the notice of filing for FAP 2A4738 (67 FR
47823). No new information or comments have been received that would
affect the agency's previous determination that there is no significant
impact on the human environment and that an environmental impact
statement is not required.
VI. Paperwork Reduction Act of 1995
This final rule contains no collection of information. Therefore,
clearance by the Office of Management and Budget under the Paperwork
Reduction Act of 1995 is not required.
VII. Objections
Any person who will be adversely affected by this regulation may
file with the Division of Dockets Management (see ADDRESSES) written or
electronic objections. Each objection shall be separately numbered, and
each numbered objection shall specify with particularity the provisions
of the regulation to which objection is made and the grounds for the
objection. Each numbered objection on which a hearing is requested
shall specifically so state. Failure to request a hearing for any
particular objection shall constitute a waiver of the right to a
hearing on that objection. Each numbered objection for which a hearing
is requested shall include a detailed description and analysis of the
specific factual information intended to be presented in support of the
objection in the event that a hearing is held. Failure to include such
a description and analysis for any particular objection shall
constitute a waiver of the right to a hearing on the objection. Three
copies of all documents are to be submitted and are to be identified
with the docket number found in brackets in the heading of this
document. Any objections received in response to the regulation may be
seen in the Division of Dockets Management between 9 a.m. and 4 p.m.,
Monday through Friday.
VIII. References
The following references have been placed on display in the
Division of Dockets Management (see ADDRESSES) and may be seen by
interested persons between 9 a.m. and 4 p.m., Monday through Friday.
1. Memorandum dated June 3, 2005, from Division of Petition
Review, Toxicology Group I, Tina Walker, to Raphael Davy, DPR,
entitled ``Safety Review of LMP-102\TM\ as an antimicrobial agent in
ready-to-eat foods, fresh meat, meat products, fresh poultry, and
poultry products.''
2. Memorandum dated April 11, 2005, from Division of Petition
Review, Chemistry Review Group, Hyoung Lee, to Regulatory Group II,
R. Davy, entitled ``FAP 2A4738 (MATS1137 M 2.3), Petition
for the use of LMP-102\TM\--a mixture of several monoclonal
bacteriophages as an antimicrobial agent in ready-to-eat meat and
poultry, Submissions of 10/25/04, 1/18/05, 1/25/05, and 2/18/05.''
3. Memorandum dated February 1, 2006, from Division of Petition
Review, Toxicology Group I, Tina Walker, to Raphael Davy, DPR,
entitled ``Addendum to the June 3, 2005 Final Toxicology Memorandum:
Additional toxicological evaluation of the potential allergenicity/
immunotoxicity of the Listera bacteriophage, LMP-102\TM\.''
4. Memorandum dated June 1, 2005, from Division of Biotechnology
and GRAS Notice Review, Negash Belay, to Raphael A. Davy, Division
of Petition Review, entitled ``Revised FAP 2A4738.''
List of Subjects in 21 CFR Part 172
Food additives, Incorporation by reference, Reporting and
recordkeeping requirements.
0
Therefore, under the Federal Food, Drug, and Cosmetic Act and under
authority delegated to the Commissioner of Food and Drugs, 21 CFR part
172 is amended as follows:
PART 172--FOOD ADDITIVES PERMITTED FOR DIRECT ADDITION TO FOOD FOR
HUMAN CONSUMPTION
0
1. The authority citation for 21 CFR part 172 continues to read as
follows:
Authority: 21 U.S.C. 321, 341, 342, 348, 371, 379e.
0
2. Section 172.785 is added to subpart H to read as follows:
Sec. 172.785 Listeria-specific bacteriophage preparation.
The additive may be safely used as an antimicrobial agent specific
for Listeria monocytogenes (L. monocytogenes) in accordance with the
following conditions:
(a) Identity. (1) The additive consists of a mixture of equal
proportions of six different individually purified lytic-type (lacking
lysogenic activity) bacteriophages (phages) specific against L.
monocytogenes.
(2) Each phage is deposited at, and assigned an identifying code
by, a scientifically-recognized culture collection center, and is made
available to FDA upon request.
(3) The additive is produced from one or more cell cultures of L.
monocytogenes in a safe and suitable nutrient medium.
(b) Specifications.
(1) The additive achieves a positive lytic result (OD600
<= 0.06) when tested
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against any of the following L. monocytogenes isolates available from
American Type Culture Collection (ATCC): ATCC 35152 (serogroup 1/2a),
ATCC 19118 (serogroup 4b), and ATCC 15313 (serogroup 1/2b). The
analytical method for determining the potency of the additive entitled
``Determination of Potency of LMP-102\TM\,'' dated October 9, 2003, and
printed by Intralytix, Inc., is incorporated by reference. The Director
of the Office of the Federal Register approves this incorporation by
reference in accordance with 5 U.S.C. 552(a) and 1 CFR part 51. You may
obtain a copy from the Office of Food Additive Safety (HFS-200), Center
for Food Safety and Applied Nutrition, Food and Drug Administration,
5100 Paint Branch Pkwy., College Park, MD 20740, or you may examine a
copy at the Center for Food Safety and Applied Nutrition's Library,
5100 Paint Branch Pkwy., College Park, MD 20740, or at the National
Archives and Records Administration (NARA). For information on the
availability of this material at NARA, call 202-741-6030, or go to:
http://www.archives.gov/federal_register/code_of_federal_regulations/ibr--locations.html.
(2) The mean phage titer of each monophage in the additive is 1 x
10\9\ plaque forming units (PFU)/ml. The analytical method for
determining phage titer entitled ``Method to Determine Lytic Activity/
Phage Titer,'' dated November 6, 2001, and printed by Intralytix, Inc.,
is incorporated by reference. Copies are available at locations cited
in paragraph (b)(1) of this section.
(3) The phages present in the preparation must not contain a
functional portion of any of the toxin-encoding sequences described in
40 CFR 725.421(d). No sequences derived from genes encoding bacterial
16S ribosomal RNA are present in the complete genomic sequence of the
phages.
(4) L. monocytogenes toxin, listeriolysin O (LLO), is not greater
than 5 hemolytic units (HU)/ml. The analytical method for determining
LLO entitled ``Quantitation of Listeriolysin O Levels in LMP-102\TM\,''
dated September 27, 2004, and printed by Intralytix, Inc., is
incorporated by reference. Copies are available at locations cited in
paragraph (b)(1) of this section.
(5) The additive is negative for L. monocytogenes. The modified
version of the U.S. Department of Agriculture's method for determining
L. monocytogenes entitled ``LMP-102\TM\ Listeria monocytogenes
Sterility Testing,'' dated May 24, 2004, and printed by Intralytix,
Inc., is incorporated by reference. Copies are available at locations
cited in paragraph (b)(1) of this section.
(6) The additive is negative for gram-positive and gram-negative
bacteria capable of growing in commonly used microbiological media
(e.g., Luria-Bertani (LB) medium), including Escherichia coli,
Salmonella species and coagulase-positive Staphylococci, as determined
by the ``Method to Determine Microbial Contamination,'' dated July 11,
2003, and printed by Intralytix, Inc., is incorporated by reference.
Copies are available at locations cited in paragraph (b)(1) of this
section.
(7) Total organic carbon (TOC) is less than or equal to 36 mg/kg.
The analytical method for determining TOC entitled ``Determination of
Total Organic Carbon by Automated Analyzer,'' dated March 30, 2001, and
printed by Intralytix, Inc., is incorporated by reference. Copies are
available at locations cited in paragraph (b)(1) of this section.
(c) Conditions of use. The additive is used in accordance with
current good manufacturing practice to control L. monocytogenes by
direct application to meat and poultry products that comply with the
ready-to-eat definition in 9 CFR 430.1. Current good manufacturing
practice is consistent with direct spray application of the additive at
a rate of approximately 1 mL of the additive per 500 cm\2\ product
surface area.
Dated: August 3, 2006.
Jeffrey Shuren,
Assistant Commissioner for Policy.
[FR Doc. E6-13621 Filed 8-17-06; 8:45 am]
BILLING CODE 4160-01-S