[Federal Register Volume 66, Number 159 (Thursday, August 16, 2001)]
[Rules and Regulations]
[Pages 42957-42962]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 01-20665]


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ENVIRONMENTAL PROTECTION AGENCY

40 CFR Part 180

[OPP-301129; FRL-6782-8]
RIN 2070-AB78


B-D-Glucuronidase from E. coli and the Genetic Material Necessary 
for its Production As a Plant Pesticide Inert Ingredient; Exemption 
from the Requirement of a Tolerance

AGENCY: Environmental Protection Agency (EPA).

ACTION: Final rule.

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SUMMARY: This regulation establishes an exemption from the requirement 
of a tolerance for residues of B-D-glucuronidase from Escherichia coli 
and the genetic material necessary for its production in or on all food 
commodities when applied/used as a plant pesticide inert ingredient. 
Monsanto submitted a petition to EPA under the Federal Food, Drug, and 
Cosmetic Act, as amended by the Food Quality Protection Act of 1996, 
requesting an exemption from the requirement of a tolerance. This 
regulation eliminates the need to establish a maximum permissible level 
for residues of B-D-glucuronidase derived from E. coli and the genetic 
material necessary for its production.

DATES: This regulation is effective August 16, 2001. Objections and 
requests for hearings, identified by docket control number (OPP-
301129), must be received by EPA, on or before October 15, 2001.

ADDRESSES: Written objections and hearing requests may be submitted by 
mail, electronically, or in person. Please follow the detailed 
instructions for each method as provided in Unit IX. of the 
SUPPLEMENTARY INFORMATION. To ensure proper receipt by EPA, your 
objections and hearing requests must identify docket control number 
(OPP-301129) in the subject line on the first page of your response.

FOR FURTHER INFORMATION CONTACT: By mail: Linda Hollis, c/o Product 
Manager (PM) 90, Biopesticides and Pollution Prevention Division 
(7511C), Environmental Protection Agency, 1200 Pennsylvania Ave., NW., 
Washington, DC 20460; telephone number: (703-308-8733); and e-mail 
address: [email protected].

SUPPLEMENTARY INFORMATION:

I. General Information

A. Does this Action Apply to Me?

    You may be affected by this action if you are an agricultural 
producer, food manufacturer, or pesticide manufacturer. Potentially 
affected categories and entities may include, but are not limited to:

------------------------------------------------------------------------
                                                          Examples of
           Categories                    NAICS            Potentially
                                                       Affected Entities
------------------------------------------------------------------------
Industry                          111                 Crop production
                                  112                 Animal production
                                  311                 Food manufacturing
                                  32532               Pesticide
                                                       manufacturing
------------------------------------------------------------------------

    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 the table could also be 
affected. The North American Industrial Classification System (NAICS) 
codes have been provided to assist you and others in determining 
whether or not this action might apply to certain entities. If you have 
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 Additional Information, Including Copies of this 
Document and Other Related Documents?

    1. Electronically. You may obtain electronic copies of this 
document, and certain other related documents that might be available 
electronically, from the EPA Internet Home Page at http://www.epa.gov/. 
To access this document, on the Home Page select ``Laws and 
Regulations,'' ``Regulations and Proposed Rules,'' and then look up the 
entry for this document under the ``Federal Register --Environmental 
Documents.'' You can also go directly to the Federal Register listings 
at http://www.epa.gov/fedrgstr/. A frequently updated electronic 
version of 40 CFR part 180 is available at http://www.access.gpo.gov/nara/cfr/cfrhtml_00/Title_40/40cfr180_00.html, a beta site currently 
under development.
    2. In person. The Agency has established an official record for 
this action under docket control number OPP-301129. The official record 
consists of the documents specifically referenced in this action, and 
other information related to this action, including any information 
claimed as Confidential Business Information (CBI). This official 
record includes the documents that are physically located in the 
docket, as well as the documents that are referenced in those 
documents. The public version of the official record does not include 
any information claimed as CBI. The public version of the official 
record, which includes printed, paper versions of any electronic 
comments submitted during an applicable comment period is available for 
inspection in the Public Information and Records Integrity Branch 
(PIRIB), Rm. 119, Crystal Mall #2, 1921 Jefferson Davis Hwy., 
Arlington, VA, from 8:30 a.m. to 4 p.m., Monday through Friday, 
excluding legal holidays. The PIRIB telephone number is (703) 305-5805.

II. Background and Statutory Findings

    In the Federal Register of May 3, 2000 (65 FR 25719) (FRL-6553-2), 
EPA issued a notice pursuant to section 408

[[Page 42958]]

of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), 
as amended by the Food Quality Protection Act (FQPA) (Public Law 104-
170) announcing the filing of a pesticide tolerance petition (PP) 
OE6066 by Monsanto. This notice included a summary of the petition 
prepared by the petitioner Monsanto. There were no comments received in 
response to the notice of filing.
    The petition requested that 40 CFR 180.1216 be amended by 
establishing an exemption from the requirement of a tolerance for 
residues of B-D-glucuronidase derived from E. coli and the genetic 
material necessary for its production.

III. Risk Assessment

    Section 408(c)(2)(A)(i) of the FFDCA allows EPA to establish an 
exemption from the requirement for a tolerance (the legal limit for a 
pesticide chemical residue in or on a food) only if EPA determines that 
the tolerance is ``safe.'' Section 408(c)(2)(A)(ii) defines ``safe'' to 
mean that ``there is a reasonable certainty that no harm will result 
from aggregate exposure to the pesticide chemical residue, including 
all anticipated dietary exposures and all other exposures for which 
there is reliable information.'' This includes exposure through 
drinking water and in residential settings, but does not include 
occupational exposure. Section 408(b)(2)(C) requires EPA to give 
special consideration to exposure of infants and children to the 
pesticide chemical residue in establishing a tolerance and to ``ensure 
that there is a reasonable certainty that no harm will result to 
infants and children from aggregate exposure to the pesticide chemical 
residue....'' Additionally, section 408(b)(2)(D) requires that the 
Agency consider ``available information'' concerning the cumulative 
effects of a particular pesticide's residues and ``other substances 
that have a common mechanism of toxicity. ''
    EPA performs a number of analyses to determine the risks from 
aggregate exposure to pesticide residues. First, EPA determines the 
toxicity of pesticides. Second, EPA examines exposure to the pesticide 
through food, drinking water, and through other exposures that occur as 
a result of pesticide use in residential settings.

IV. Toxicological Profile

    Consistent with section 408(b)(2)(D) of FFDCA, EPA has reviewed the 
available scientific data and other relevant information in support of 
this action and considered its validity, completeness, and reliability 
and the relationship of this information to human risk. EPA has also 
considered available information concerning the variability of the 
sensitivities of major identifiable subgroups of consumers, including 
infants and children.
    Several types of data are required for proteinaceous plant-
pesticide ingredients to provide a reasonable certainty that no harm 
will result from the aggregate exposure to these proteins. The 
information is intended to show that a proteinaceous plant-pesticide 
behaves as would be expected of a typical dietary protein, is not 
structurally related to any known food allergen or protein toxin, and 
does not display any oral toxicity when administered at high doses. 
These data consist of an in vitro digestion assay, amino acid sequence 
homology comparisons and an acute oral toxicity test. The acute oral 
toxicity test is done at a maximum hazard dose using purified protein 
of the plant-pesticide as a test substance.
    EPA believes that protein instability in digestive fluids and the 
lack of adverse effects using the maximum hazard dose approach in 
general eliminate the need for longer-term testing of Bt protein plant-
pesticide ingredients. Dosing of these animals with the maximum hazard 
dose, along with the product characterization data should identify 
potential toxins and allergens, and provide an effective means to 
determine the safety of these protein. The adequacy of the current 
testing requirements was discussed at the June 7, 200 Scientific 
Advisory Panel (SAP) meeting.
    Toxicity studies submitted in support of this tolerance exemption 
included the following:
    Acute oral toxicity (44988-01). One hundred CD-1 albino mice 
divided into groups of 10 male and 10 female each were treated with 1.0 
milligrams/kilograms (mg/kg), 10.0 mg/kg or 100.0 mg/kg of the test 
substance (GUS protein) or 100.0 mg/kg control substance (bovine serum 
albumin). Treatment was administered by oral gavage at 33 ml/kg body 
weight. Three unscheduled deaths occurred on day 5. These animals were 
in each of the following groups: 100.0 mg control substance, 10.0 mg or 
100.0 mg test substance. Since the deaths appeared in both control and 
test substance groups, there were no abnormal observations upon gross 
necropsy of the animals that died prematurely, the deaths were not dose 
related and only three deaths were seen in the one hundred animals of 
the study, it appears that the deaths were not related to the test 
substance. Minor weight loss was recorded in all groups and routine, 
minor organ abnormalities were also seen in both the treated and 
control groups during gross necropsy at schedules sacrifice. Based upon 
the data, there were no significant adverse effects reported upon 
dosing mice with up to 100.0 mg/kg body weight GUS protein.
    In vitro digestibility (449394-07). This study was performed on B-
D-glucuronidase purified from E. coli strain K12 engineered to express 
the GUS protein to determine the digestive fate of the protein in 
simulated gastric and intestinal fluid. The data submitted indicate 
that the GUS protein is broken down rapidly with incubation in 
simulated gastric fluids but is relatively stable in simulated 
intestinal fluids. GUS protein enzymatic activity rapidly disappears 
after incubation in simulated gastric fluid (2 minutes, the first 
timepoint examined). GUS protein also disappears when examined 
immunologically by western blot as quickly as 15 seconds after 
incubation in simulated gastric fluid. Although still degraded, GUS 
protein is more stable to intestinal digestion disappearing by 240 
minutes by western blot yet still being detected by enzymatic activity 
(decreased about 90%) at this same time point. These results suggest 
that the protein breaks down in the human digestive tract as expected 
of a dietary protein and will not likely pose a risk in foods as part 
of the human diet.
    The B-D-glucuronidase which is the subject of this rule is a 
protein originally isolated from E. coli and introduced into plants to 
serve as a transformation marker. GUS (B-D-glucuronide 
glucouronosohydrolase; E.C.3.2.1.31) from E. coli is a homotetrameric 
enzyme with an individual monomeric weight of 68kD. Individual subunits 
do not have enzymatic activity. GUS has a pH optima near 7.0 and 
maintains enzymatic activity for approximately 2 hours at 55 degrees C 
but is rapidly degraded at 60 degrees C. This bacterial enzymatic 
activity is ubiquitous in the digestive system of humans and other 
vertebrates. As a protein component of the normal microflora of the 
intestinal tract, there will be no change in exposure from the presence 
of this protein as a transformation marker. In addition, other types of 
GUS enzyme are present in the liver, spleen, kidneys, salivary glands, 
breast milk and a variety of other tissues in humans, other vertebrates 
and a number of invertebrates. While these proteins have similar 
activity, the mammalian safety data generated to date has been 
specifically for the E. coli derived GUS so the present tolerance is 
limited to that form.

[[Page 42959]]

    Allergenic responses are very unlikely to occur and the Agency is 
currently unaware of any allergic reactions to this protein. The 
highest activity of the mammalian GUS protein is found primarily in the 
liver and kidneys. However, activity has also been seen in the spleen, 
breast milk, adrenal gland and alimentary tract. GUS protein is also 
found in many other bacteria besides E. coli and is also present in the 
environment (Ref. 1). The GUS protein is used as a marker gene during 
the plant transformation process in the development of genetically 
modified plants. During the plant transformation process, the GUS 
protein serves as an identifier which enables the separation of 
transformed plant cells containing an added gene from those plant cells 
that have failed to take up or maintain the additional gene of 
interest. Thus, the GUS protein allows a cell expressing that marker 
gene to be readily identified.
    The mammalian health and safety of the GUS protein is based on its 
ubiquitous presence in the digestive system of humans and other tissues 
(Refs. 2 and 6), as well as its presence in anaerobic bacteria (Ref. 
3), and other environmental bacteria (Ref. 7). Further, the mammalian 
health and safety of the GUS protein is based on the long history of 
safe consumption of the protein in the human and domestic animal food 
supply (Ref. 2), and in the tissue of various plant species from which 
foods such as potatoes, apples, almonds, rye, sugar beets, etc., are 
derived (Refs. 4, 8, and 9). There have been no reports of adverse 
effects to humans or domestic animals from the consumption of the GUS 
protein. Toxicity studies conducted in support of this tolerance 
exemption indicated a lack of toxicity of the E. coli derived GUS 
protein in mice. Moreover, in vitro digestibility studies further 
validate earlier findings that the E. coli derived GUS protein is 
broken down under conditions in mammalian digestive fluids. Therefore, 
the Agency concludes that the risk to humans when consuming foods 
containing the GUS protein is minimal to non-existent. The lack of heat 
stability of the GUS protein suggests that cooking foods would 
eliminate the protein activity (Ref. 5). Further, the data submitted 
suggest that upon ingestion, the GUS protein rapidly degrades in the 
digestive tract thus posing no risks of adverse effects to humans.
    The genetic material necessary for the production of the plant 
pesticide inert ingredient are the nucleic acids (DNA) which comprise 
genetic material encoding the protein and their regulatory regions. 
Regulatory regions are the genetic material that control the expression 
of the genetic material encoding the proteins, such as promoters, 
terminators, and enhancers. DNA is common to all forms of plant and 
animal life and the Agency knows of no adverse effects related to their 
consumption as a component of food. These ubiquitous nucleic acids as 
they appear in the subject plant pesticide inert ingredient have been 
adequately characterized by the applicant. The EPA concludes that no 
mammalian toxicity is anticipated from dietary exposure to the genetic 
material necessary for the production of the GUS protein.

V. Aggregate Exposures

    In examining aggregate exposure, FFDCA section 408 directs EPA to 
consider available information concerning exposures from the pesticide 
residue in food and all other non-occupational exposures, including 
drinking water from ground water or surface water and exposure through 
pesticide use in gardens, lawns, or buildings (residential and other 
indoor uses).

A. Dietary Exposure

    1. Food. The use of B-D-glucuronidase from E. coli and the genetic 
material necessary for its production as a plant pesticide inert 
ingredient will not result in new dietary exposure to human health or 
to the environment. The GUS protein is ubiquitous in the digestive 
tract of humans, other vertebrate, invertebrates, bacteria, in the 
environment and in foods in the human and domestic animal diet. 
Exposure from this protein has not posed any unreasonable risk or 
health concerns. The lack of mammalian toxicity and the rapid 
degradation of the protein by the stomach digestion of the GUS protein 
provide a scientific rationale for exempting from the requirement of a 
tolerance B-D-glucuronidase and the genetic material necessary for its 
production when used as a plant pesticide inert ingredient (Ref. 10). 
Moreover, there is no evidence indicating adverse effects due to 
aggregate exposure of the GUS protein through dietary, non-food oral, 
dermal and inhalation.
    2. Drinking water exposure. Potential exposures in drinking water 
is negligible. Because GUS protein is contained within the cells of the 
plant and is naturally degraded upon plant senescence, the likely 
transfer of the GUS protein to drinking water is minimal to non-
existent.

B. Other Non-Occupational Exposure

    Other non-occupational exposure of B-D-glucuronidase via 
residential and indoor uses e.g., uses around homes, parks, recreation 
areas, athletic fields and golf courses will be minimal to non-existent 
as the GUS protein is contained within the plant cells.
    1. Dermal exposure. Due to the nature of the GUS protein contained 
within the plant cells as part of the plant pesticide inert ingredient, 
exposure through any route (i.e., dermal, respiratory) other than 
dietary is unlikely to occur. Physical contact with the plant or raw 
agricultural food from the plant may present some limited opportunity 
for dermal exposure. However, on a per person basis, the potential 
amounts involved in this exposure is negligible in comparison to 
exposure through the dietary route.
    2. Inhalation exposure. The occurrence of respiratory exposure of 
the GUS protein contained within the plant cells is negligible in 
comparison to potential exposure through the dietary route.

VI. Cumulative Effects

    E. coli derived B-D-glucuronidase enzyme and its gene is present as 
part of a ubiquitous organism in the digestive systems of humans and 
other vertebrates. Based on the lack of mammalian toxicity and the long 
history of safe consumption of the protein in the human and domestic 
animal food supply and the rapid degradation of the protein in the 
digestive tract, no cumulative effects with other substances are 
expected.

VII. Determination of Safety for U.S. Population, Infants and 
Children

    For the U.S. population, including infants and children, B-D-
glucuronidase from E. coli and the genetic material necessary for its 
production as a plant pesticide inert ingredient has no known or 
reported adverse effects. The Agency's conclusions are based on the 
extensive use and experience with the GUS protein including the long 
history of safe consumption of the protein in the human and domestic 
animal food supply, the lack of mammalian toxicity associated with the 
protein, the rapid degradation of the protein by the stomach digestion 
prior to passage to the intestinal tract, along with no reported 
adverse effects due to aggregate exposure through the dietary, non-food 
oral, dermal and inhalation routes. Therefore, based on all available 
information, the EPA concludes that there is reasonable certainty that 
no harm will result from aggregate

[[Page 42960]]

exposure of the U.S. population, including infants and children, to the 
GUS protein when used as a plant pesticide inert ingredient, as 
expressed in plants in or on all food commodities.

VIII. Other Considerations

A. Endocrine Disruptors

    EPA is required under the FFDCA, as amended by FQPA, to develop a 
screening program to determine whether certain substances (including 
all pesticide active and other ingredient) ``may have an effect in 
humans that is similar to an effect produced by a naturally occurring 
estrogen, or other such endocrine effects as the Administrator may 
designate.'' Following the recommendations of the Endocrine Disruptor 
Screening and Testing Advisory Committee (EDSTAC), EPA determined that 
there was scientific basis for including, as part of the program, the 
androgen-and thyroid hormone systems, in addition to the estrogen 
hormone system. EPA also adopted EDSTAC's recommendation that the 
program include evaluations of potential effects in wildlife. For 
pesticide chemicals, EPA will use FIFRA and to the extent that FFDCA 
authority to require the wildlife evaluations. As the science develops 
and resources allow, screening of additional hormone systems may be 
added to the Endocrine Disruptor Screening Program (EDSP).
    When the appropriate screening and or testing protocols being 
considered under the Agency's Endocrine Disruptor Screening Program 
have been developed, B-D-glucuronidase from E. coli and the genetic 
material necessary for its production may be subjected to additional 
screening and/or testing to better characterize effects related to 
endocrine disruption. Based on the weight of the evidence of available 
data, no endocrine system-related effects have been identified.

B. Analytical Method(s)

    The Agency proposes to establish an exemption from the requirement 
of a tolerance without any numerical limitation for the reasons stated 
above. For the same reasons, the Agency has concluded that an 
analytical method is not required for enforcement purposes for B-D-
glucuronidase from E. coli and the genetic material necessary for its 
production.

C. Codex Maximum Residue Level

    There are no Codex Maximum Residue Levels (MRL's) established for 
residues of B-D-glucuronidase from E. coli and the genetic material 
necessary for its production.

IX. Objections and Hearing Requests

    Under section 408(g) of the FFDCA, as amended by the FQPA, any 
person may file an objection to any aspect of this regulation and may 
also request a hearing on those objections. The EPA procedural 
regulations which govern the submission of objections and requests for 
hearings appear in 40 CFR part 178. Although the procedures in those 
regulations require some modification to reflect the amendments made to 
the FFDCA by the FQPA of 1996, EPA will continue to use those 
procedures, with appropriate adjustments, until the necessary 
modifications can be made. The new section 408(g) provides essentially 
the same process for persons to ``object'' to a regulation for an 
exemption from the requirement of a tolerance issued by EPA under new 
section 408(d), as was provided in the old FFDCA sections 408 and 409. 
However, the period for filing objections is now 60 days, rather than 
30 days.

A. What Do I Need to Do to File an Objection or Request a Hearing?

    You must file your objection or request a hearing on this 
regulation in accordance with the instructions provided in this unit 
and in 40 CFR part 178. To ensure proper receipt by EPA, you must 
identify docket control number OPP-301140 in the subject line on the 
first page of your submission. All requests must be in writing, and 
must be mailed or delivered to the Hearing Clerk on or before October 
15, 2001.
    1. Filing the request. Your objection must specify the specific 
provisions in the regulation that you object to, and the grounds for 
the objections (40 CFR 178.25). If a hearing is requested, the 
objections must include a statement of the factual issues(s) on which a 
hearing is requested, the requestor's contentions on such issues, and a 
summary of any evidence relied upon by the objector (40 CFR 178.27). 
Information submitted in connection with an objection or hearing 
request may be claimed confidential by marking any part or all of that 
information as CBI. Information so marked will not be disclosed except 
in accordance with procedures set forth in 40 CFR part 2. A copy of the 
information that does not contain CBI must be submitted for inclusion 
in the public record. Information not marked confidential may be 
disclosed publicly by EPA without prior notice.
    Mail your written request to: Office of the Hearing Clerk (1900), 
Environmental Protection Agency, Ariel Rios Bldg., 1200 Pennsylvania 
Ave., NW., Washington, DC 20460. You may also deliver your request to 
the Office of the Hearing Clerk in Rm. C400, Waterside Mall, 401 M St., 
SW., Washington, DC 20460. The Office of the Hearing Clerk is open from 
8 a.m. to 4 p.m., Monday through Friday, excluding legal holidays. The 
telephone number for the Office of the Hearing Clerk is (202) 260-4865.
    2. Tolerance fee payment. If you file an objection or request a 
hearing, you must also pay the fee prescribed by 40 CFR 180.33(i) or 
request a waiver of that fee pursuant to 40 CFR 180.33(m). You must 
mail the fee to: EPA Headquarters Accounting Operations Branch, Office 
of Pesticide Programs, P.O. Box 360277M, Pittsburgh, PA 15251. Please 
identify the fee submission by labeling it ``Tolerance Petition Fees.''
    EPA is authorized to waive any fee requirement ``when in the 
judgement of the Administrator such a waiver or refund is equitable and 
not contrary to the purpose of this subsection.'' For additional 
information regarding the waiver of these fees, you may contact James 
Tompkins by phone at (703) 305-5697, by e-mail at [email protected], 
or by mailing a request for information to Mr. Tompkins at Registration 
Division (7505C), Office of Pesticide Programs, Environmental 
Protection Agency, 1200 Pennsylvania Ave., NW., Washington, DC 20460.
    If you would like to request a waiver of the tolerance objection 
fees, you must mail your request for such a waiver to: James Hollins, 
Information Resources and Services Division (7502C), Office of 
Pesticide Programs, Environmental Protection Agency, 1200 Pennsylvania 
Ave., NW., Washington, DC 20460.
    3. Copies for the Docket. In addition to filing an objection or 
hearing request with the Hearing Clerk as described in Unit IX.A., you 
should also send a copy of your request to the PIRIB for its inclusion 
in the official record that is described in Unit I.B.2. Mail your 
copies, identified by docket number OPP-301140, to: Public Information 
and Records Integrity Branch, Information Resources and Services 
Division (7502C), Office of Pesticide Programs, Environmental 
Protection Agency, 1200 Pennsylvania Ave., NW., Washington, DC 20460. 
In person or by courier, bring a copy to the location of the PIRIB 
described in Unit I.B.2. You may also send an electronic copy of your 
request via e-mail to: [email protected]. Please use an ASCII file 
format and avoid the use of special characters and any form of 
encryption. Copies of electronic objections and hearing requests will 
also be accepted on disks in WordPerfect 6.1/8.0 or ASCII file

[[Page 42961]]

format. Do not include any CBI in your electronic copy. You may also 
submit an electronic copy of your request at many Federal Depository 
Libraries.

B. When Will the Agency Grant a Request for a Hearing?

    A request for a hearing will be granted if the Administrator 
determines that the material submitted shows the following: There is a 
genuine and substantial issue of fact; there is a reasonable 
possibility that available evidence identified by the requestor would, 
if established resolve one or more of such issues in favor of the 
requestor, taking into account uncontested claims or facts to the 
contrary; and resolution of the factual issues(s) in the manner sought 
by the requestor would be adequate to justify the action requested (40 
CFR 178.32).

X. References

    1. Fuchs, R.L. and Astwood, J.D. 1996. Allergenicity assessment of 
foods derived from genetically modified plants. Food Technology 50: 83-
88.
    2. Gilissen, L. J. W., P. L. J. Metz, W. J. Stiekema and J. P. Nap. 
1998. Biosafety of E. coli b-glucuronidase (GUS) in plants. Trans. Res. 
7:157-163.
    3. Hawkesworth, G., B.S. Draser and M.J. Hill. 1971. Intestinal 
bacteria and the hydrolysis of glycosidic bonds. J. Med. Microbiol. 
4:451-9.
    4. Hodal, L. A. Bochardt, J.E. Nielsen, O. Mattsson, and F.T. Okk. 
1992. Detection, expression and specific elimination of endogenous b-
glucuronidase activity in transgenic and non-transgenic plants. Plant 
Science. 87:115-22.
    5. Jefferson, R.A. and K.J. Wilson. 1991. The GUS gene fusion 
system. Plant Mol. Biol. Manual B14:1-33.
    6. Jefferson, R. A., T. A. Kavanagh, and M. W. Bevan. 1986. B-
Glucuronidase from Escherichia coli as a gene-fusion marker. Proc. 
Natl. Acad. Sci. USA 83: 8447-8451.
    7. Levvy, G.A. and C.A. Marsh. 1959. Preparation and properties of 
b-glucuronidase. Adv. Carbohydrate Chem. 14:381-428.
    8. Schulz, M. and G. Weissenbock. 1987. Dynamics of the tissue-
specific metabolism of lutolin glucuronides in the mesophyll of rye 
primary leaves (Secale cereale) Z. Naturforsh. 43c: 187-93.
    9. Wozniak, C.A. and L.D. Owens. 1994. Native B-glucuronidase 
activity in sugarbeet (Beta vulgaris). Physiol. Plant. 90: 763-771.
    10. Kough, J. 2001. Note to file: Safety assessment of B-
glucuronidase as a worker gene/inert ingredient.

XI. Regulatory Assessment Requirements

    This final rule establishes an exemption from the tolerance 
requirement under FFDCA section 408(d) in response to a petition 
submitted to the Agency. The Office of Management and Budget (OMB) has 
exempted these types of actions from review under Executive Order 
12866, entitled Regulatory Planning and Review (58 FR 51735, October 4, 
1993). This final rule does not contain any information collections 
subject to OMB approval under the Paperwork Reduction Act (PRA), 44 
U.S.C. 3501 et seq., or impose any enforceable duty or contain any 
unfunded mandate as described under Title II of the Unfunded Mandates 
Reform Act of 1995 (UMRA) (Public Law 104 -4). Nor does it require any 
prior consultation as specified by Executive Order 13084, entitled 
Consultation and Coordination with Indian Tribal Governments (63 FR 
27655, May 19, 1998); special considerations as required by Executive 
Order 12898, entitled Federal Actions to Address Environmental Justice 
in Minority Populations and Low-Income Populations (59 FR 7629, 
February 16, 1994); or require OMB review or any Agency action under 
Executive Order 13045, entitled Protection of Children from 
Environmental Health Risks and Safety Risks (62 FR 19885, April 23, 
1997). This action does not involve any technical standards that would 
require Agency consideration of voluntary consensus standards pursuant 
to section 12(d) of the National Technology Transfer and Advancement 
Act of 1995 (NTTAA), Public Law 104-113, section 12(d) (15 U.S.C. 272 
note). Since tolerances and exemptions that are established on the 
basis of a petition under FFDCA section 408(d), such as the exemption 
in this final rule, do not require the issuance of a proposed rule, the 
requirements of the Regulatory Flexibility Act (RFA) (5 U.S.C. 601 et 
seq.) do not apply. In addition, the Agency has determined that this 
action will not have a substantial direct effect on States, on the 
relationship between the national government and the States, or on the 
distribution of power and responsibilities among the various levels of 
government, as specified in Executive Order 13132, entitled Federalism 
(64 FR 43255, August 10, 1999). Executive Order 13132 requires EPA to 
develop an accountable process to ensure ``meaningful and timely input 
by State and local officials in the development of regulatory policies 
that have federalism implications.'' ``Policies that have federalism 
implications'' is defined in the Executive Order to include regulations 
that have ``substantial direct effects on the States, on the 
relationship between the national government and the States, or on the 
distribution of power and responsibilities among the various levels of 
government.'' This final rule directly regulates growers, food 
processors, food handlers and food retailers, not States. This action 
does not alter the relationships or distribution of power and 
responsibilities established by Congress in the preemption provisions 
of FFDCA section 408(n)(4).

XII. Submission to Congress and the Comptroller General

    The Congressional Review Act, 5 U.S.C. 801 et seq., as added by the 
Small Business Regulatory Enforcement Fairness Act of 1996, generally 
provides that before a rule may take effect, the agency promulgating 
the rule must submit a rule report, which includes a copy of the rule, 
to each House of the Congress and to the Comptroller General of the 
United States. EPA will submit a report containing this rule and other 
required information to the U.S. Senate, the U.S. House of 
Representatives, and the Comptroller General of the United States prior 
to publication of this final rule in the Federal Register. This final 
rule is not a ``major rule'' as defined by 5 U.S.C. 804(2).

List of Subjects in 40 CFR Part 180

    Environmental protection, Administrative practice and procedure, 
Agricultural commodities, Pesticides and pests, Reporting and 
recordkeeping requirements.


    Dated: July 11, 2001.

Marcia E. Mulkey,
Director, Office of Pesticide Programs.

    Therefore, 40 CFR chapter I is amended as follows:

PART 180--[AMENDED]

    1. The authority citation for part 180 continues to read as 
follows:

    Authority: 21 U.S.C. 321(q), 346(a) and 371.

    2. Section 180.1216 is added to subpart D to read as follows:


Sec. 180.1216  B-D-glucuronidase from E. coli and the genetic material 
necessary for its production as a plant-pesticide inert ingredient; 
exemption from the requirement of a tolerance.

    An exemption from the requirement of a tolerance is established for 
residues of B-D-glucuronidase from E. coli and the genetic material 
necessary for its production when used as a plant-

[[Page 42962]]

pesticide inert ingredient in or on all food commodities. Genetic 
material necessary for the production means both: Genetic material that 
encodes a substance or leads to the production of a substance; and 
regulatory regions. It does not include non-coding, non-expressed 
nucleotide sequences. Regulatory region means genetic material that 
controls the expression of the genetic material that encodes a 
pesticidal substance or leads to the production of a pesticidal 
substance. Examples of regulatory regions include, but are not limited 
to, promoters, enhancers, and terminators.

[FR Doc. 01-20665 Filed 8-15-01; 8:45 a.m.]
BILLING CODE 6560-50-S