[Federal Register Volume 68, Number 112 (Wednesday, June 11, 2003)]
[Notices]
[Pages 34959-34962]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 03-14327]



[[Page 34959]]

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ENVIRONMENTAL PROTECTION AGENCY

[OPP-2003-0165; FRL-7306-4]


Notice of Filing a Pesticide Petition to Establish a Tolerance 
for a Certain Pesticide Chemical in or on Food

AGENCY: Environmental Protection Agency (EPA).

ACTION: Notice.

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SUMMARY: This notice announces the initial filing of a pesticide 
petition proposing the establishment of regulations for residues of an 
inert ingredient hygromycin B phosphotransferase (APH4) marker protein 
and the genetic material necessary for its production in or on various 
food commodities.

DATES: Comments, identified by docket ID number OPP-2003-0165, must be 
received on or before July 11, 2003.

ADDRESSES:  Comments may be submitted electronically, by mail, or 
through hand delivery/courier. Follow the detailed instructions as 
provided in Unit I. of the SUPPLEMENTARY INFORMATION.

FOR FURTHER INFORMATION CONTACT: Leonard Cole, Biopesticides and 
Pollution Prevention Division (7511C), Office of Pesticide Programs, 
Environmental Protection Agency, 1200 Pennsylvania Ave., NW., 
Washington, DC 20460-0001; telephone number: (703) 305-5412; e-mail 
address: [email protected].

SUPPLEMENTARY INFORMATION:

I. General Information

A. Does this Action Apply to Me?

    You may be potentially affected by this action if you are an 
agricultural producer, food manufacturer, or pesticide manufacturer. 
Potentially affected entities may include, but are not limited to:
    [sbull] Crop production (NAICS 111)
    [sbull] Animal production (NAICS 112)
    [sbull] Food manufacturing (NAICS 311)
    [sbull] Pesticide manufacturing (NAICS 32532)
    This listing is not intended to be exhaustive, but rather provides 
a guide for readers regarding entities likely to be affected by this 
action. Other types of entities not listed in this unit could also be 
affected. The North American Industrial Classification System (NAICS) 
codes have been provided to assist you and others in determining 
whether this action might apply to certain entities. If you have any 
questions regarding the applicability of this action to a particular 
entity, consult the person listed under FOR FURTHER INFORMATION 
CONTACT.

B. How Can I Get Copies of this Document and Other Related Information?

    1. Docket. EPA has established an official public docket for this 
action under docket identification (ID) number OPP-2003-0165. The 
official public docket consists of the documents specifically 
referenced in this action, any public comments received, and other 
information related to this action. Although a part of the official 
docket, the public docket does not include Confidential Business 
Information (CBI) or other information whose disclosure is restricted 
by statute. The official public docket is the collection of materials 
that is available for public viewing at the Public Information and 
Records Integrity Branch (PIRIB), Rm. 119, Crystal Mall 2, 
1921 Jefferson Davis Hwy., Arlington, VA. This docket facility is open 
from 8:30 a.m. to 4 p.m., Monday through Friday, excluding legal 
holidays. The docket telephone number is (703) 305-5805.
    2. Electronic access. You may access this Federal Register document 
electronically through the EPA Internet under the ``Federal Register'' 
listings at http://www.epa.gov/fedrgstr/.
    An electronic version of the public docket is available through 
EPA's electronic public docket and comment system, EPA Dockets. You may 
use EPA Dockets at http://www.epa.gov/edocket/ to submit or view public 
comments, access the index listing of the contents of the official 
public docket, and to access those documents in the public docket that 
are available electronically. Although not all docket materials may be 
available electronically, you may still access any of the publicly 
available docket materials through the docket facility identified in 
Unit I.B.1. Once in the system, select ``search,'' then key in the 
appropriate docket ID number.
    Certain types of information will not be placed in the EPA Dockets. 
Information claimed as CBI and other information whose disclosure is 
restricted by statute, which is not included in the official public 
docket, will not be available for public viewing in EPA's electronic 
public docket. EPA's policy is that copyrighted material will not be 
placed in EPA's electronic public docket but will be available only in 
printed, paper form in the official public docket. To the extent 
feasible, publicly available docket materials will be made available in 
EPA's electronic public docket. When a document is selected from the 
index list in EPA Dockets, the system will identify whether the 
document is available for viewing in EPA's electronic public docket. 
Although not all docket materials may be available electronically, you 
may still access any of the publicly available docket materials through 
the docket facility identified in Unit I.B.1. EPA intends to work 
towards providing electronic access to all of the publicly available 
docket materials through EPA's electronic public docket.
    For public commenters, it is important to note that EPA's policy is 
that public comments, whether submitted electronically or in paper, 
will be made available for public viewing in EPA's electronic public 
docket as EPA receives them and without change, unless the comment 
contains copyrighted material, CBI, or other information whose 
disclosure is restricted by statute. When EPA identifies a comment 
containing copyrighted material, EPA will provide a reference to that 
material in the version of the comment that is placed in EPA's 
electronic public docket. The entire printed comment, including the 
copyrighted material, will be available in the public docket.
    Public comments submitted on computer disks that are mailed or 
delivered to the docket will be transferred to EPA's electronic public 
docket. Public comments that are mailed or delivered to the docket will 
be scanned and placed in EPA's electronic public docket. Where 
practical, physical objects will be photographed, and the photograph 
will be placed in EPA's electronic public docket along with a brief 
description written by the docket staff.

C. How and to Whom Do I Submit Comments?

    You may submit comments electronically, by mail, or through hand 
delivery/courier. To ensure proper receipt by EPA, identify the 
appropriate docket ID number in the subject line on the first page of 
your comment. Please ensure that your comments are submitted within the 
specified comment period. Comments received after the close of the 
comment period will be marked ``late.'' EPA is not required to consider 
these late comments. If you wish to submit CBI or information that is 
otherwise protected by statute, please follow the instructions in Unit 
I.D. Do not use EPA Dockets or e-mail to submit CBI or information 
protected by statute.
    1. Electronically. If you submit an electronic comment as 
prescribed in this unit, EPA recommends that you include your name, 
mailing address, and an e-mail address or other contact information in 
the body of your

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comment. Also include this contact information on the outside of any 
disk or CD ROM you submit, and in any cover letter accompanying the 
disk or CD ROM. This ensures that you can be identified as the 
submitter of the comment and allows EPA to contact you in case EPA 
cannot read your comment due to technical difficulties or needs further 
information on the substance of your comment. EPA's policy is that EPA 
will not edit your comment, and any identifying or contact information 
provided in the body of a comment will be included as part of the 
comment that is placed in the official public docket, and made 
available in EPA's electronic public docket. If EPA cannot read your 
comment due to technical difficulties and cannot contact you for 
clarification, EPA may not be able to consider your comment.
    i. EPA Dockets. Your use of EPA's electronic public docket to 
submit comments to EPA electronically is EPA's preferred method for 
receiving comments. Go directly to EPA Dockets at http://www.epa.gov/edocket, and follow the online instructions for submitting comments. 
Once in the system, select ``search,'' and then key in docket ID number 
OPP-2003-0165. The system is an ``anonymous access'' system, which 
means EPA will not know your identity, e-mail address, or other contact 
information unless you provide it in the body of your comment.
    ii. E-mail. Comments may be sent by e-mail to [email protected], 
Attention: Docket ID Number OPP-2003-0165. In contrast to EPA's 
electronic public docket, EPA's e-mail system is not an ``anonymous 
access'' system. If you send an e-mail comment directly to the docket 
without going through EPA's electronic public docket, EPA's e-mail 
system automatically captures your e-mail address. E-mail addresses 
that are automatically captured by EPA's e-mail system are included as 
part of the comment that is placed in the official public docket, and 
made available in EPA's electronic public docket.
    iii. Disk or CD ROM. You may submit comments on a disk or CD ROM 
that you mail to the mailing address identified in Unit I.C.2. These 
electronic submissions will be accepted in WordPerfect or ASCII file 
format. Avoid the use of special characters and any form of encryption.
    2. By mail. Send your comments to: Public Information and Records 
Integrity Branch (PIRIB) (7502C), Office of Pesticide Programs (OPP), 
Environmental Protection Agency, 1200 Pennsylvania Ave., NW., 
Washington, DC 20460-0001, Attention: Docket ID Number OPP-2003-0165.
    3. By hand delivery or courier. Deliver your comments to: Public 
Information and Records Integrity Branch (PIRIB), Office of Pesticide 
Programs (OPP), Environmental Protection Agency, Rm. 119, Crystal Mall 
2, 1921 Jefferson Davis Hwy., Arlington, VA, Attention: Docket 
ID Number OPP-2003-0165. Such deliveries are only accepted during the 
docket's normal hours of operation as identified in Unit I.B.1.

D. How Should I Submit CBI to the Agency?

    Do not submit information that you consider to be CBI 
electronically through EPA's electronic public docket or by e-mail. You 
may claim information that you submit to EPA as CBI by marking any part 
or all of that information as CBI (if you submit CBI on disk or CD ROM, 
mark the outside of the disk or CD ROM as CBI and then identify 
electronically within the disk or CD ROM the specific information that 
is CBI). Information so marked will not be disclosed except in 
accordance with procedures set forth in 40 CFR part 2.
    In addition to one complete version of the comment that includes 
any information claimed as CBI, a copy of the comment that does not 
contain the information claimed as CBI must be submitted for inclusion 
in the public docket and EPA's electronic public docket. If you submit 
the copy that does not contain CBI on disk or CD ROM, mark the outside 
of the disk or CD ROM clearly that it does not contain CBI. Information 
not marked as CBI will be included in the public docket and EPA's 
electronic public docket without prior notice. If you have any 
questions about CBI or the procedures for claiming CBI, please consult 
the person listed under FOR FURTHER INFORMATION CONTACT.

E. What Should I Consider as I Prepare My Comments for EPA?

    You may find the following suggestions helpful for preparing your 
comments:
    1. Explain your views as clearly as possible.
    2. Describe any assumptions that you used.
    3. Provide copies of any technical information and/or data you used 
that support your views.
    4. If you estimate potential burden or costs, explain how you 
arrived at the estimate that you provide.
    5. Provide specific examples to illustrate your concerns.
    6. Make sure to submit your comments by the deadline in this 
notice.
    7. To ensure proper receipt by EPA, be sure to identify the docket 
ID number assigned to this action in the subject line on the first page 
of your response. You may also provide the name, date, and Federal 
Register citation.

II. What Action is the Agency Taking?

    EPA has received a pesticide petition as follows proposing the 
establishment and/or amendment of regulations for residues of a certain 
pesticide chemical in or on various food commodities under section 408 
of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a. 
EPA has determined that this petition contains data or information 
regarding the elements set forth in FFDCA section 408(d)(2); however, 
EPA has not fully evaluated the sufficiency of the submitted data at 
this time or whether the data support granting of the petition. 
Additional data may be needed before EPA rules on the petition.

List of Subjects

    Environmental protection, Agricultural commodities, Feed additives, 
Food additives, Pesticides and pests, Reporting and recordkeeping 
requirements.

    Dated: May 29, 2003.
Janet L. Andersen,
Director, Biopesticides and Pollution Prevention Division, Office of 
Pesticide Programs.

Summary of Petition

    The petitioner summary of the pesticide petition is printed below 
as required by FFDCA section 408(d)(3). The summary of the petition was 
prepared by the petitioner and represents the view of the petitioner. 
The petition summary announces the availability of a description of the 
analytical methods available to EPA for the detection and measurement 
of the pesticide chemical residues or an explanation of why no such 
method is needed.

Syngenta Seeds, Inc.

PP 3G6590

    EPA has received pesticide petition (PP 3G6590) from Syngenta 
Seeds, Inc., P.O. Box 12257, 3054 Cornwallis Road, Research Triangle 
Park, NC 27709-2257, proposing pursuant to section 408(d) of the 
Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to 
amend 40 CFR part 180 to establish an exemption from the requirement of 
a tolerance for the plant-pesticide inert ingredient hygromycin B 
phosphotransferase (APH4) marker protein and the genetic material

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necessary for its production in or on cotton.
    Pursuant to section 408(d)(2)(A)(i) of the FFDCA, as amended, 
Syngenta Seeds, Inc. has submitted the following summary of 
information, data, and arguments in support of their pesticide 
petition. This summary was prepared by Syngenta Seeds, Inc. and EPA has 
not fully evaluated the merits of the pesticide petition. The summary 
may have been edited by EPA if the terminology used was unclear, the 
summary contained extraneous material, or the summary unintentionally 
made the reader conclude that the findings reflected EPA's position and 
not the position of the petitioner.

A. Product Name and Proposed Use Practices

    Hygromycin B phosphotransferase (APH4) marker protein is proposed 
for use as a plant-incorporated protectant formulation inert 
ingredient. APH4 protein is an aminocyclitol phosphotransferase that 
catalyzes the phosphorylation of hygromycin and closely related 
aminoglycoside antibiotics. Expression of the APH4 gene in plant cells 
allows for growth and selection of transformed cells in the presence of 
hygromycin B. APH4 has no insecticidal activity.

B. Product Identity/Chemistry

    1. Identity of the pesticide and corresponding residues. The aph4 
gene in event COT102 cotton plants was derived from a plasmid harbored 
by a hygromycin-resistant isolate of E. coli, and encodes a 341 amino-
acid enzyme, hygromycin B phosphotransferase (APH4). Hygromycin B 
phosphotransferases with significant homology to the APH4 protein in 
event COT102 plants have also been identified in other microbes 
including Streptomyces hygroscopicus, the source of hygromycin B.
    APH4 has a molecular weight of ca. 42,000 and catalyzes the 
phosphorylation of the 4-hydroxyl group on the hyosamine moiety of 
hygromycin B, thereby inactivating it. The enzyme has a narrow range of 
substrates, in that it phosphorylates hygromycin B, hygromycin 
B2 and the closely related antibiotics destomycin A and 
destomycin B, but does not phosphorylate other aminocyclitol or 
aminoglycoside antibiotics including neomycin, streptomycin, 
gentamicin, kanamycin, spectinomycin, tobramycin, and amikacin. 
Hygromycin B is not used in human clinical therapy, but is principally 
used as an antihelminthic agent in swine and poultry feeds.
    2. Magnitude of residue at the time of harvest and method used to 
determine the residue. A determination of the magnitude of residue at 
harvest is not required for residues exempt from tolerances. However, 
the petitioner has provided data on the quantity of APH4 protein 
measured in various plant parts including seeds of VIP3A cotton, as 
measured by enzyme linked immunosorbent assay (ELISA). APH4 was either 
not detectable in most COT102 plant tissues or the levels were too low 
to quantify. Pollen was the only tissue in which quantifiable levels 
were measured.
    3. A statement of why an analytical method for detecting and 
measuring the levels of the pesticide residue are not needed. An 
analytical method is not required because this petition requests an 
exemption from tolerances. However, the petitioner has submitted an 
analytical method for detection of the APH4 protein by ELISA analysis.

C. Mammalian Toxicological Profile

    Syngenta Seeds is providing the results of a mammalian toxicology 
study, in vitro digestibility study, and bioinformatics evaluations 
conducted on the selectable marker protein APH4. These studies, 
summarized herein, demonstrate the lack of toxicity of the APH4 protein 
following acute oral exposure to mice, rapid degradation of APH4 upon 
exposure to simulated gastric and intestinal fluids, and the lack of 
amino acid sequence similarity of the APH4 protein to proteins known to 
be mammalian toxins or human allergens.
    When proteins are toxic, they are known to act via acute mechanisms 
and at very low doses (Ref. 1). Therefore, when a protein demonstrates 
no acute oral toxicity in high-dose testing using a standard laboratory 
mammalian test species, this supports the determination that the 
protein will be non-toxic to humans and other mammals, and will not 
present a hazard under any realistic exposure scenario, including long-
term exposures.
    Because it is not possible to extract sufficient APH4 protein from 
transformed plants for toxicology studies, APH4 protein was produced in 
recombinant E. coli by over-expressing the same aph4 gene that was 
introduced into VIP3A cotton event COT102. The aph4 gene was cloned 
into the inducible, over-expression pET-3a[reg] vector (Novagen, 
Madison, WI) in E. coli BL21DE3pLysS. The APH4 protein, as encoded in 
this vector, was identical in amino acid sequence to that encoded by 
the plant transformation vector, pCOT1, except for an additional 11 
amino acids from the T7 TagTM and three amino acids from the 
vector polylinker. Following purification from E. coli, dialysis and 
lyophilization, the resulting sample, designated Test Substance APH4-
0102, was estimated by ELISA to contain ca. 42.6% APH4 protein by 
weight. The test material was confirmed to be enzymatically active.
    An acute mouse oral toxicity study was conducted at the Syngenta 
Central Toxicology Laboratory (Alderley Park, Macclesfield, Cheshire, 
UK) according to OPPTS Harmonized Guideline 870.1100. Test substance 
APH4-0102 (see above description of test substance) was administered to 
five male and five female mice strain Alderley Park albino mouse 
(APfCD-1); 8-9 weeks old via a gavage dose of 1,828 milligrams/kilogram 
(mg/kg) body weight. The test substance contained ca. 42.6% APH4 
protein by weight. Therefore, the mice received ca. 779 mg APH4/kg body 
weight. A negative control group (5 mice/sex) concurrently received the 
dosing vehicle alone, a suspension of 1% methylcellulose, at the same 
dosing volume as used for the test material mixture. Food was provided 
ad libitum, except during the ca. 1-hour prior to dosing, when the 
animals were fasted. Water was provided ad libitum throughout the 
study. Observations for mortality and clinical/behavioral signs of 
toxicity were made at least twice on the day of dosing, and at least 
once daily thereafter for 14 days. Detailed clinical observations were 
made for each animal at each observation time. Body weights were 
recorded daily and food consumption was recorded weekly. Surviving 
animals were euthanized 14 days post dosing and subjected to gross 
necropsy. Organ weights (brain, liver with gall bladder, kidneys and 
spleen) were recorded and principal tissues were processed for 
microscopic examination.
    No mortalities occurred during the study, and no clinical signs of 
toxicity were observed in either the test or control groups. There were 
no treatment-related effects on body weight, food consumption, or organ 
weights, nor were any treatment-related effects observed following 
macroscopic or microscopic examination. APH4-0102 is not acutely toxic 
to mice. There is no evidence of toxicity of the test substance at 
1,828 mg APH4-0102/kg body weight, representing ca. 779 mg APH4 
protein/kg body weight. The estimated LD50 value for pure 
APH4 protein in male and female mice is >779 mg/kg body weight, the 
single dose tested.
    The APH4 protein shows no homology to proteins known to be

[[Page 34962]]

mammalian toxins or human allergens; is not derived from a source known 
to produce allergens; is not targeted to a cellular pathway for 
glycosylation in the plant; and is rapidly degraded upon exposure to 
simulated gastric and intestinal.
    The genetic material necessary for the production of APH4 as an 
inert ingredient are the nucleic acids (DNA) which comprise genetic 
material encoding this protein and its regulatory regions. ``Regulatory 
regions'' are the genetic material that control the expression of the 
genetic material encoding the protein, such as promoters, terminators, 
and enhancers. DNA is common to all forms of plant and animal life and 
the Agency has previously stated that they are not aware of an instance 
where these nucleic acids have been associated with toxic effects 
related to their consumption as a component of food. These ubiquitous 
nucleic acids, as they appear in the subject inert ingredient, have 
been adequately characterized. Therefore, no mammalian toxicity is 
anticipated from dietary exposure to the genetic material necessary for 
the production of the subject inert plant pesticidal ingredient.

D. Aggregate Exposure

    1. Dietary exposure--i. Food. Derivatives of cottonseed (e.g., 
refined cottonseed oil) and fiber (e.g., linters, which are essentially 
100% cellulose) are used in some food products. However, APH4 was not 
detected in most of the samples of COT102-derived cottonseed analyzed 
or any of the cotton fiber samples analyzed. In the few cottonseed 
samples in which APH4 was detectable, the quantities were below the 
limit of quantification (<137 ng APH4/g fresh wt; <150 ng APH4/g dry 
wt). It is expected that any trace quantities of APH4 in cottonseed 
will be eliminated by standard seed processing methods. As demonstrated 
by the analysis of cottonseed products for VIP3A protein, no VIP3A was 
detected in refined cottonseed oil from COT102-derived plants, despite 
the presence of ca. 3 micrograms VIP3A/g seed (fresh or dry wt.). 
Additionally, no protein of any kind was detected in the same sample of 
refined cottonseed oil. It can be concluded that APH4, as produced in 
COT102-derived cotton plants, does not pose a risk of becoming 
allergenic via food, because there will be no exposure via food. 
Additionally, the APH4 protein shows no amino acid sequence homology to 
known allergens; is not derived from a source known to produce 
allergens; is not targeted to a cellular pathway for glycosylation in 
the plant; and is rapidly degraded upon exposure to simulated gastric 
and intestinal.
    ii. Drinking water. No exposure to the APH4 and the genetic 
material necessary for its production as an inert ingredient via 
drinking water are expected. The protein is incorporated into the plant 
and will therefore not be available to drinking water sources.
    2. Non-dietary exposure. Non-dietary exposure is not anticipated, 
due to the proposed use pattern of the product. Exposure via dermal or 
inhalation routes is unlikely because the inert ingredient is contained 
within plant cells. However, if exposure were to occur by non-dietary 
routes, no risk would be expected because the APH4 protein is not toxic 
to mammals.

E. Cumulative Exposure

    Because there is no indication of mammalian toxicity to the APH4 
protein, it is reasonable to conclude that there are no cumulative 
effects for this inert ingredient.

F. Safety Determination

    1.  U.S. population. The lack of mammalian toxicity at high levels 
of exposure to the APH4 protein demonstrates the safety of the product 
at levels well above possible maximum exposure levels anticipated via 
consumption of processed food products produced from VIP3A cotton. 
Moreover, little to no human dietary exposure to APH4 protein is 
expected to occur via VIP3A cotton. Due to the lack of toxicity of the 
APH4 protein and its very low potential for allergenicity, dietary 
exposure is not anticipated to pose any harm for the U.S. population. 
No special safety provisions are applicable for consumption patterns or 
for any population sub-groups.
    2. Infants and children. Syngenta has evaluated the acute toxicity 
data generated on APH4, the lack of homology to known allergens or 
toxins, and the limited exposure to this protein based on the residue 
profile and limited number of food/feed products resulting from cotton 
and has determined that there is ample evidence to indicate a 
reasonable certainty of no harm to infants and children as a result of 
the use of this product.

G. Effects on the Immune and Endocrine Systems

    The inert ingredient APH4 is a protein, derived from sources that 
are not known to exert an influence on the endocrine or immune systems.

H. Existing Tolerances

    The registrant is not aware of any known existing tolerances or 
exemptions for APH4 and the genetic material necessary for its 
production as an inert ingredient.

I. International Tolerances

    The registrant is not aware that any Codex maximum residue levels 
exist for the APH4 protein and the genetic material necessary for its 
production.

J. Reference

    1. Sjoblad, R.D., J.T. McClintock and R. Engler (1992) 
Toxicological considerations for protein components of biological 
pesticide products. Regulatory Toxicol. Pharmacol. 15: 3-9.

[FR Doc. 03-14327 Filed 6-10-03; 8:45 am]
BILLING CODE 6560-50-S