[Federal Register Volume 62, Number 122 (Wednesday, June 25, 1997)]
[Notices]
[Pages 34271-34276]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 97-16659]
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ENVIRONMENTAL PROTECTION AGENCY
[PF-739; FRL-5721-7]
Notice of Filing of Pesticide Petitions
AGENCY: Environmental Protection Agency (EPA).
ACTION: Notice.
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SUMMARY: This notice announces the initial filing of pesticide
petitions proposing the establishment of regulations for residues of
certain pesticide chemicals in or on various food commodities.
DATES: Comments, identified by the docket control number PF-739, must
be received on or before July 25, 1997.
ADDRESSES: By mail submit written comments to: Public Information and
Records Integrity Branch, Information Resources and Services Division
(7506C), Office of Pesticides Programs, Environmental Protection
Agency, 401 M St., SW., Washington, DC 20460. In person bring comments
to: Rm. 1132, CM #2, 1921 Jefferson Davis Highway, Arlington, VA.
Comments and data may also be submitted electronically by following
the instructions under ``SUPPLEMENTARY INFORMATION.'' No confidential
business information should be submitted through e-mail.
Information submitted as a comment concerning this document may be
claimed confidential by marking any part or all of that information as
``Confidential Business Information'' (CBI). CBI should not be
submitted through e-mail. Information marked as CBI will not be
disclosed except in accordance with procedures set forth in 40 CFR part
2. A copy of the comment 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. All written
comments will be available for public inspection in Rm. 1132 at the
address given above, from 8:30 a.m. to 4 p.m., Monday through Friday,
excluding legal holidays.
FOR FURTHER INFORMATION CONTACT: The regulatory action leaders listed
in the table below:
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Office location/
Product Manager telephone number Address
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Sheryl Reilly (PM 90)......... Rm. 5-W29, 5th Floor, 2800 Jefferson
CS-1, 703-308-8265 e- Davis Hwy.,
mail: Arlington, VA
[email protected] 22202
.epa.gov
Mike Mendelsohn (PM 90)....... Rm. 5-W44, 5th Floor, Do.
CS-1, 703-308-8715 e-
mail:
mendelsohn.mike@epama
Linda Hollis (PM 90).......... Rm 5-J, 5th Floor, CS- Do.
1, 703-308-8733 e-
mail:
hollis.linda@epamail.
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SUPPLEMENTARY INFORMATION: EPA has received pesticide petitions as
follows proposing the establishment and/or amendment of regulations for
residues of certain pesticide chemicals in or on various food
commodities under section 408 of the Federal Food, Drug, and Comestic
Act (FFDCA), 21 U.S.C. 346a. EPA has determined that these petitions
contain data or information regarding the elements set forth in section
408(d)(2); however, EPA has not fully evaluated the sufficiency of the
submitted data at this time or whether the data supports granting of
the petition. Additional data may be needed before EPA rules on the
petition.
The official record for this notice of filing, as well as the
public version, has been established for this notice of filing under
docket control number [PF-739] (including comments and data submitted
electronically as described below). A public version of this record,
including printed, paper versions of electronic comments, which does
not include any information claimed as CBI, is available for inspection
from 8:30 a.m. to 4 p.m., Monday through Friday, excluding legal
holidays. The official record is located at the address in
``ADDRESSES'' at the beginning of this document.
Electronic comments can be sent directly to EPA at:
opp-docket@epamail.epa.gov
Electronic comments must be submitted as an ASCII file avoiding the
use of special characters and any form of encryption. Comment and data
will also be accepted on disks in
[[Page 34272]]
Wordperfect 5.1 file format or ASCII file format. All comments and data
in electronic form must be identified by the docket number (insert
docket number) and appropriate petition number. Electronic comments on
notice may be filed online at many Federal Depository Libraries.
List of Subjects
Environmental protection, Agricultural commodities, Food additives,
Feed additives, Pesticides and pests, Reporting and recordkeeping
requirements.
Dated: June 12, 1997.
Janet L. Andersen,
Director, Biopesticides and Pollution Prevention Division, Office of
Pesticide Programs.
Summaries of Petitions
Petitioner summaries of the pesticide petitions are printed below
as required by section 408(d)(3) of the FFDCA. The summaries of the
petitions were prepared by the petitioners and represent the views of
the petitioners. EPA is publishing the petition summaries verbatim
without editing them in any way. 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.
1. Kemira Agro Oy
PP 7F4137
A. Proposed Use Practices
Registration of PRIMASTOP containing Gliocladium catenulatum Strain
J1446 is being proposed for the following sites: Vegetables, herbs and
spices, ornamentals, tree and shrub seedlings, turf, home and garden.
PRIMASTOP is used for the control of damping-off, seed rot, root
and stem rot, and wilt diseases caused by Rhizoctonia, Pythium,
Phytophthora, Fusarium, Didymella, Botrytis, Verticillium, Alternaria,
Cladosporium, Helminthosporium, Penicillium and Plicaria on vegetables,
herbs, ornamentals and tree and forest seedlings grown in greenhouse or
outdoors.
PRIMASTOP can be incorporated in the growth substrate as a dry
powder or as an aqueous suspension or applied by drenching, spraying or
dipping.
1. Incorporation into potting media. The recommended rate for
incorporation of PRIMASTOP in potting media is 5 to 30 oz/
yd3 (0.2 to 1 g/L) of growing media. If the incorporation is
done with the aqueous suspension, mix 3.5 oz (100 g) of PRIMASTOP
powder in 1.0 gallon (or 4 L) of water and carefully mix the suspension
with the growth substrate (1.5-8.5 gal/yd3). Incorporation
of PRIMASTOP can be followed with a drench application within 2 to 6
weeks.
2. Drench application. Drenching treatment can be done using a 0.2-
0.5% suspension. Seedling trays or beds can be drenched with PRIMASTOP
at the recommended rate of 2 to 10 oz./100 ft2 (5-25 g/
m2). Drenching at sowing is recommended for vegetables
(except for tomato and leek), herbs and ornamentals (except pansy)
grown in peat or soil mixture. Drenching after emergence is recommended
for tomato, leek, pansy and all seedlings grown in rockwool, such as
cucumbers. Repeat treatment at transplanting.
3. Foliar spray. PRIMASTOP can be sprayed or spread on the plant
stems or foliar parts for control of Didymella or Botrytis with an
aqueous suspension. Recommended concentration is up to 5%.
4. Treatment of cuttings, bulbs or tubers. Cuttings, bulbs or
tubers can be dipped in or sprayed with PRIMASTOP suspensionn before
planting or storage. The product can also be incorporated in the
storage mixture, such as sand or peat at a rate up to 1 g/L.
B. Product Identity/Chemistry
1. Identity of pesticide and corresponding residues. The active
ingredient in Primastop is Gliocladium catenulatum Strain J1446. The
mechanism by which Gliocladium catenulatum Strain J1446 controls
diseases appears to be enzymatic. Gliocladium catenulatum Strain J1446
does not produce toxins or antibiotics. Further, Gliocladium
catenulatum Strain J1446 is a naturally occurring microorganism.
Gliocladium catenulatum is widespread in the environment.
2. Magnitude of residue anticipated at the time of harvest and
method used to determine the residue. No residues of Gliocladium
catenulatum Strain J1446 are anticipated in treated crops at harvest.
Subdivision M - Series 153A-3(a) indicates that ``if Tier I toxicology
tests indicate no toxic or other harmful properties, then no residue
data would be indicated.'' Studies with Gliocladium catenulatum Strain
J1446 demonstrated low mammalian toxicity. No pathogenicity or
infectivity was observed in any of the tests conducted with Gliocladium
catenulatum Strain J1446. Further, Gliocladium catenulatum Strain J1446
is a naturally occurring microorganism. Gliocladium catenulatum is
widespread in the environment.
3. Statement of why an analytical method for detecting and
measuring the levels of the pesticide residue are not needed.
Subdivision M - Series 153A-3(a) indicates that ``if Tier I toxicology
tests indicate no toxic or other harmful properties , then no residue
data would be indicated and thus a recommendation for an exemption from
the requirement of a tolerance can be made.'' Studies with Gliocladium
catenulatum Strain J1446 demonstrated low mammalian toxicity. No
pathogenicity or infectivity was observed in any of the tests conducted
with Gliocladium catenulatum Strain J1446. Further, Gliocladium
catenulatum Strain J1446 is a naturally occurring microorganism.
Gliocladium catenulatum is widespread in the environment.
C. Health and Safety
Kemira Agro Oy conducted the required toxicology studies to support
its petition for an exemption from the requirement of tolerance and
associated registrations. The studies conducted indicate a low
mammalian toxicity for Gliocladium catenulatum Strain J1446. No
pathogenicity or infectivity was observed in any of the tests conducted
with Gliocladium catenulatum Strain J1446. The mechanism by which
Gliocladium catenulatum Strain J1446 controls diseases appears to be
enzymatic. Gliocladium catenulatum Strain J1446 does not produce toxins
or antibiotics.
Toxicology data in support of the exemption from the requirement of
a tolerance for Gliocladium catenulatum Strain J1446 included studies
with the cell mass (technical) and with the formulated product as
follows:
1. Acute toxicity and/or pathogenicity.-- a. Gliocladium
catenulatum Strain J1446 Cell Mass (Technical). i. Acute oral toxicity
and pathogenicity in rats (acute oral LD50 > 4.04 to 5.86
x 108 cfu/kg, clearance: < 3="" days).="" ii.="" acute="" pulmonary="" toxicity/pathogenicity="" in="" rats="" (acute="" pulmonary="">50 > 6.60 to 7.98 x 108 cfu/kg, clearance: < 7="" days).="" iii.="" acute="" intraperitoneal="" toxicity/pathogenicity="" in="" rats="" (acute="" intraperitoneal="">50 > 4.2 x 108 cfu/kg, clearance: < 7="" days).="" b.="" gliocladium="" catenulatum="" strain="" j1446="" formulation="" (primastop).="" i.="" acute="" oral="">50 toxicity in rats (> 2,000 mg/kg, EPA
toxicity category III).
ii. Acute dermal LD50 toxicity in rats (>2,000 mg/kg,
EPA toxicity category III).
[[Page 34273]]
iii. Acute dermal irritation in rabbits (minimal dermal irritant,
EPA toxicity category IV).
iv. Acute inhalation LC50 toxicity in rats (> 5.57 mg/L,
EPA toxicity category V).
v. Primary eye irritation (minimal eye irritant, EPA toxicity
category IV).
vi. Skin sensitization (sensitizer).
vii. No hypersensitivity effects have been observed.
c. The inert ingredients contained in the Gliocladium catenulatum
Strain J1446 formulation, Primastop, are all minimal risk (List 4)(59
FR 49400, September 28, 1994).
2. Genotoxicity. Subdivision M Guidelines do not require the
conduct of genotoxicity studies to support the registration of a
microbial pest control agent, such as Gliocladium catenulatum Strain
J1446.
3. Reproductive and developmental toxicity. Subdivision M
Guidelines do not require the conduct of reproductive and developmental
toxicity studies to support the registration of a microbial pest
control agent, such as Gliocladium catenulatum Strain J1446.
4. Subchronic toxicity. Subdivision M Guidelines do not require the
conduct of subchronic toxicity studies to support the registration of a
microbial pest control agent, such as Gliocladium catenulatum Strain
J1446.
5. Chronic toxicity. Subdivision M Guidelines do not require the
conduct of chronic toxicity studies to support the registration of a
microbial pest control agent, such as Gliocladium catenulatum Strain
J1446.
Sufficient data exist to assess the hazards of Gliocladium
catenulatum Strain J1446 and to make a determination on aggregate
exposure, consistent with section 408(c)(2), for the exemptions from
the requirement of a tolerance. The exposures, including dietary
exposure, and risks associated with establishing the requested
exemption from the requirement of a tolerance follows.
D. Threshold Effects
Gliocladium catenulatum is a naturally occurring microorganism that
is widespread in the environment. Both the cell mass (technical) and
the formulation of Gliocladium catenulatum Strain J1446 demonstrated
low toxicity. No pathogenicity or infectivity was observed in any of
the tests conducted with Gliocladium catenulatum Strain J1446. No
threshold effects were observed or are anticipated for Gliocladium
catenulatum Strain J1446.
E. Non-threshold Effects
Gliocladium catenulatum is a naturally occurring microorganism that
is widespread in the environment. Both the cell mass (technical) and
formulation of Gliocladium catenulatum Strain J1446 demonstrated low
toxicity. No pathogenicity or infectivity was observed in any of the
tests conducted with Gliocladium catenulatum Strain J1446. Non-
threshold effects were not observed nor are any anticipated for
Gliocladium catenulatum Strain J1446.
F. Aggregate Exposure
Gliocladium catenulatum is naturally occurring and widespread in
the environment. The low toxicity and non-pathogenicity/infectivity of
Gliocladium catenulatum Strain J1446 is demonstrated by the data
summarized above. The product will be applied by incorporation into
growing media and/or by drenching at seeding or in the early growing
stages of the treated plants.
1. Dietary exposure-- a. food. It is not anticipated that residues
of Gliocladium catenulatum Strain J1446 will occur in treated raw
agricultural commodities.
b. Drinking water. It is not anticipated that residues of
Gliocladium catenulatum Strain J1446 will occur in drinking water.
2. Non-dietary exposure. The potential for non-occupational, non-
dietary exposure to the general population is not expected to be
significant.
G. Cumulative Exposure
There is no anticipated potential for cumulative effects of
Gliocladium catenulatum Strain J1446 and other substances that have a
common mechanism of toxicity. Clearance of Gliocladium catenulatum
Strain J1446 from test species was < 3="" days="" in="" two="" studies="" and="">< 7="" days="" in="" a="" third="" study.="" toxic="" effects="" produced="" by="" gliocladium="" catenulatum="" strain="" j1446="" should="" not="" be="" cumulative="" with="" those="" of="" any="" other="" chemical="" compounds.="" h.="" determination="" of="" safety="" for="" u.s.="" population="" gliocladium="" catenulatum="" strain="" j1446="" is="" a="" naturally="" occurring="" microorganism.="" gliocladium="" catenulatum="" is="" widespread="" in="" the="" environment.="" the="" low="" toxicity="" of="" gliocladium="" catenulatum="" strain="" j1446="" is="" demonstrated="" by="" the="" data="" summarized="" above.="" based="" on="" this="" information,="" the="" aggregate="" exposure="" to="" gliocladium="" catenulatum="" strain="" j1446="" over="" a="" lifetime="" should="" not="" pose="" appreciable="" risks="" to="" human="" health.="" there="" is="" a="" reasonable="" certainty="" that="" no="" harm="" will="" result="" from="" aggregate="" exposure="" to="" gliocladium="" catenulatum="" strain="" j1446="" residues.="" exempting="" gliocladium="" catenulatum="" strain="" j1446="" from="" the="" requirement="" of="" a="" tolerance="" should="" be="" considered="" safe="" and="" pose="" insignificant="" risk.="" i.="" determination="" of="" safety="" for="" infants="" and="" children="" the="" toxicity="" and="" exposure="" data="" are="" sufficiently="" complete="" to="" adequately="" address="" the="" potential="" for="" additional="" sensitivity="" of="" infants="" and="" children="" to="" residues="" of="" gliocladium="" catenulatum="" strain="" j1446.="" there="" is="" a="" reasonable="" certainty="" that="" no="" harm="" will="" result="" to="" infants="" and="" children="" from="" aggregate="" exposure="" to="" gliocladium="" catenulatum="" strain="" j1446="" residues.="" j.="" estrogenic="" effects="" no="" specific="" tests="" have="" been="" conducted="" with="" gliocladium="" catenulatum="" strain="" j1446="" to="" determine="" whether="" it="" may="" have="" an="" effect="" in="" humans="" that="" is="" similar="" to="" an="" effect="" produced="" by="" a="" naturally="" occurring="" estrogen="" or="" other="" endocrine="" effects.="" however,="" it="" is="" not="" likely="" that="" gliocladium="" catenulatum="" strain="" j1446="" would="" have="" estrogen="" or="" endocrine="" effects="" because:="" 1.="" it="" is="" a="" naturally="" occurring="" microorganism.="" gliocladium="" catenulatum="" is="" widespread="" in="" the="" environment.="" 2.="" it="" has="" demonstrated="" low="" mammalian="" toxicity.="" 3.="" no="" pathogenicity="" or="" infectivity="" was="" observed="" in="" any="" of="" the="" tests="" conducted="" with="" gliocladium="" catenulatum="" strain="" j1446.="" 4.="" the="" mechanism="" by="" which="" gliocladium="" catenulatum="" strain="" j1446="" controls="" diseases="" appears="" to="" be="" enzymatic.="" 5.="" gliocladium="" catenulatum="" strain="" j1446="" does="" not="" produce="" toxins="" or="" antibiotics.="" k.="" existing="" tolerances="" no="" tolerances="" or="" exemptions="" from="" the="" requirement="" of="" tolerance="" have="" been="" established="" or="" applied="" for="" domestically="" or="" internationally="" other="" that="" subject="" petition.="" l.="" environmental="" fate="" environmental="" fate="" data="" are="" not="" required="" to="" support="" the="" registration="" of="" a="" biopesticide="" unless="" results="" from="" tier="" i="" studies="" indicate="" that="" risks="" would="" be="" expected="" from="" use="" of="" the="" product.="" gliocladium="" catenulatum="" gstrain="" j1446="" is="" a="" naturally="" occurring="" microorganism.="" gliocladium="" catenulatum="" is="" widespread="" in="" the="" environment.="" extensive="" literature="" searches="" revealed="" an="" absence="" of="" any="" ecological="" effects="" or="" environmental="" fate="" [[page="" 34274]]="" data="" from="" gliocladium="" catenulatum.="" (sheryl="" reilly)="" 2.="" monsanto="" company="" pp="" 6e4657="" a.="" background="" information="" and="" use="" profile="" the="" development="" of="" plant="" varieties="" containing="" useful="" new="" traits="" introduced="" by="" plant="" genetic="" engineering="" such="" as="" insect="" protection="" depends="" upon="" an="" effective="" means="" to="" select="" for="" the="" rare="" transformed="" plant="" cells="" containing="" the="" added="" gene.="" for="" example,="" a="" gene="" required="" for="" the="" production="" of="" an="" insecticidal="" protein="" in="" the="" plant="" tissue="" cannot="" be="" efficiently="" selected="" for="" several="" weeks="" after="" the="" transformation="" event="" as="" it="" does="" not,="" itself,="" provide="" a="" readily="" selectable="" property="" to="" the="" cell="" which="" carries="" it.="" regenerating="" each="" cell="" from="" that="" transformation="" experiment="" to="" test="" for="" the="" presence="" of="" the="" gene="" would="" be="" both="" impractical="" and="" prohibitory,="" as="" the="" frequency="" that="" transformed="" cells="" are="" obtained="" is="" often="" as="" low="" as="" 1="" in="" 10,000="" or="" 1="" in="" 100,000="" of="" the="" cells="" treated="" (fraley="" et="" al.,="" 1984).="" therefore,="" a="" selectable="" marker="" gene="" and="" a="" selective="" agent="" are="" used="" to="" identify="" the="" rare="" transformed="" cells="" for="" regeneration.="" a="" selectable="" marker="" gene="" allows="" a="" cell="" expressing="" that="" marker="" gene="" to="" grow="" in="" the="" presence="" of="" a="" selective="" agent="" by="" inactivating="" or="" neutralizing="" the="" agent="" which="" would="" otherwise="" inhibit="" the="" growth="" or="" kill="" the="" cell.="" the="" marker="" gene="" also="" permits="" tracking="" of="" linked="" traits="" that="" are="" difficult="" to="" identify="" at="" the="" cellular="" or="" whole="" plant="" level.="" for="" insect-protected="" corn="" plants,="" the="" gox="" gene="" was="" used="" as="" a="" selectable="" marker="" gene="" conferring="" tolerance="" to="" glyphosate.="" the="" glyphosate="" oxidoreductase="" (gox)="" gene="" from="" achromobacter="" sp.="" strain="" lbaa="" (new="" genus="" ochrobactrum="" anthropi)="" produces="" a="" protein="" (gox)="" which="" degrades="" glyphosate.="" the="" gox="" protein="" confers="" tolerance="" to="" glyphosate="" and="" provides="" a="" selectable="" marker="" used="" in="" initial="" laboratory="" stages="" of="" plant="" cell="" selection="" to="" identify="" plant="" cells="" containing="" agronomic="" genes="" of="" interest="" such="" as="" the="" cryia(b)="" gene="" which="" imparts="" protection="" from="" certain="" lepidopteran="" insect="" pests.="" b.="" risk="" assessment="" and="" statutory="" findings="" toxicology="" profile="" 1.="" data="" summary.="" monsanto="" company="" has="" requested="" an="" exemption="" from="" the="" requirement="" of="" a="" tolerance="" for="" glyphosate="" oxidoreductase="" (gox)="" as="" a="" plant="" pesticide="" formulation="" inert="" ingredient.="" included="" in="" the="" monsanto="" submission="" to="" the="" epa="" were="" several="" toxicology="" studies="" in="" support="" of="" the="" gox="" protein="" as="" a="" pesticide="" formulation="" inert="" ingredient.="" the="" gox="" protein="" used="" in="" these="" studies="" was="" produced="" in="" an="" e.="" coli="" over-expression="" system="" and="" partially="" purified.="" the="" gox="" protein="" expressed="" in="" e.="" coli="" was="" characterized="" and="" shown="" to="" be="" substantially="" equivalent="" to="" the="" gox="" expressed="" in="" insect-protected="" corn="" where="" it="" was="" utilized="" as="" a="" selectable="" marker="" protein.="" the="" following="" mammalian="" toxicity="" studies="" have="" been="" conducted="" to="" support="" this="" exemption="" from="" the="" requirement="" of="" a="" tolerance:="" a.="" a="" mouse="" acute="" oral="" gavage="" study="" was="" performed="" in="" which="" the="" no-="" observed-effect-level="" (noel)="" for="" toxicity="" of="" gox="" protein="" administered="" as="" a="" single="" dose="" was="" considered="" to="" be="" 100="" milligrams="" per="" kilogram="" (mg/="" kg)="" (the="" highest="" tested="" target="" dose).="" the="" protein="" was="" administered="" by="" gavage="" to="" three="" groups="" of="" male="" and="" female="" mice="" at="" target="" dose="" levels="" of="" 1,="" 10,="" and="" 100="" mg/kg="" body="" weight.="" appropriate="" hollow="" vector="" and="" vehicle="" controls="" were="" used.="" mice="" were="" observed="" twice="" daily="" for="" signs="" of="" toxicity="" and="" food="" consumption="" was="" recorded="" daily.="" food="" and="" water="" were="" provided="" ad="" libitum.="" all="" animals="" were="" sacrificed="" on="" post-dosing="" day="" seven="" and="" subjected="" to="" gross="" necropsy.="" approximately="" 40="" tissues="" were="" collected="" and="" saved="" for="" each="" animal="" in="" the="" test.="" there="" were="" no="" statistically="" significant="" differences="" in="" body="" weight,="" cumulative="" body="" weight="" or="" food="" consumption="" between="" the="" controls="" or="" gox="" protein="" treated="" groups.="" no="" grossly="" observable="" pathologic="" changes="" were="" observed="" in="" mice="" at="" necropsy="" that="" were="" related="" to="" treatment.="" when="" proteins="" are="" toxic,="" they="" are="" known="" to="" act="" via="" acute="" mechanisms="" and="" at="" very="" low="" dose="" levels="" (sjoblad,="" et="" al.,="" 1992).="" the="" acute="" oral="" toxicity="" data="" submitted="" support="" the="" prediction="" that="" the="" gox="" protein="" is="" non-toxic="" to="" humans.="" b.="" an="" in="" vitro="" digestive="" fate="" study="" of="" the="" gox="" protein="" in="" simulated="" gastric="" and="" intestinal="" fluids="" demonstrated="" rapid="" protein="" degradation.="" in="" gastric="" fluid,="" the="" gox="" protein="" degraded="" extremely="" rapidly;="" more="" than="" 90%="" of="" the="" initially="" added="" gox="" protein="" degraded="" after="" 15="" seconds="" incubation="" as="" detected="" by="" western="" blot="" analysis.="" gox="" enzymatic="" activity="" also="" dissipated="" readily;="" more="" than="" 96%="" of="" the="" added="" gox="" activity="" dissipated="" after="" one="" minute="" incubation,="" the="" earliest="" time="" point="" measured.="" in="" intestinal="" fluid,="" gox="" protein="" degraded="" rapidly;="" more="" than="" 90%="" of="" the="" initially="" added="" gox="" protein="" degraded="" after="" 30="" seconds="" incubation="" as="" detected="" by="" western="" blot="" analysis.="" gox="" enzymatic="" activity="" also="" dissipated="" readily;="" more="" than="" 95%="" of="" the="" added="" gox="" enzymatic="" activity="" dissipated="" after="" 60="" minute="" incubation.="" the="" difference="" in="" dissipation="" of="" the="" enzymatic="" activity="" of="" gox="" when="" compared="" to="" detection="" by="" western="" blot="" analysis="" suggests="" the="" antigenic="" sites="" on="" the="" gox="" protein="" for="" the="" particular="" antibody="" used="" in="" this="" study="" were="" more="" sensitive="" to="" proteolytic="" degradation="" in="" simulated="" intestinal="" fluid="" under="" these="" conditions="" than="" is="" the="" functional="" active="" site="" of="" the="" gox="" protein.="" the="" gox="" protein="" degraded="" readily,="" though,="" as="" assessed="" by="" both="" western="" blot="" analysis="" and="" enzymatic="" activity.="" the="" results="" of="" this="" study="" established="" that="" the="" gox="" protein="" and="" its="" associated="" functional="" activity="" will="" be="" efficiently="" degraded="" upon="" exposure="" to="" gastric="" and="" intestinal="" fluids="" in="" the="" mammalian="" digestive="" tract.="" known="" protein="" allergens="" are="" often="" resistant="" to="" digestion.="" c.="" a="" homology="" assessment="" of="" the="" amino="" acid="" sequence="" of="" the="" gox="" protein="" has="" been="" performed="" comparing="" this="" protein="" to="" known="" allergens="" or="" gliadin="" proteins.="" monsanto="" has="" searched="" the="" amino="" acid="" sequences="" of="" the="" 219="" allergens="" present="" in="" public="" domain="" genetic="" databases="" (genbank,="" embl,="" pir,="" and="" swissprot)="" for="" similarity="" to="" the="" amino="" acid="" sequences="" of="" the="" gox="" protein="" using="" the="" fasta="" computer="" program="" (pearson="" and="" lipman,="" 1988).="" monsanto="" concludes="" (i)="" that="" the="" gox="" gene="" introduced="" does="" not="" encode="" a="" known="" allergen,="" and="" (ii)="" that="" the="" introduced="" gox="" protein="" does="" not="" share="" immunologically="" significant="" sequences="" with="" known="" allergens.="" the="" gox="" protein="" is="" produced="" at="" low="" levels="" (ppm)="" by="" insect-protected="" corn="" plants="" and="" is="" contained="" within="" the="" cells="" of="" the="" corn="" plant.="" in="" documentation="" provided="" to="" the="" agency,="" the="" range="" of="" gox="" protein="" levels="" in="" insect-protected="" corn="" line="" samples="" as="" assessed="" using="" a="" validated="" elisa="" specific="" to="" the="" gox="" protein="" ranged="" from="">< 1.8="" to="" 19.32="">g/g fresh weight (fwt) in leaf tissue, < 1.5="" to="" 11.7="">g/g fwt in grain, and < 0.6="" to="" 2.46="">g/g fwt in whole
plant tissue. Western blot analysis indicated that the GOX protein was
not present in corn pollen.
The genetic material encoding the GOX protein and the regulatory
regions associated with the gene have been well characterized. Nucleic
acids (DNA) is common to all forms of plant and animal life and there
are no known instances of toxic effects related to their consumption.
No mammalian toxicity is expected from dietary exposure to the genetic
material necessary for the production of the GOX protein in corn.
In summary, the safety of the GOX protein to mammals, including
humans, was confirmed by demonstrating the rapid degradation of the GOX
protein
[[Page 34275]]
under conditions which simulate mammalian digestion, by establishing
the lack of toxicity to rodents in an acute gavage study and by
establishing the lack of allergenic concerns.
2. Acute toxicity. An acute mouse gavage study with GOX protein was
performed to directly assess potential acute toxicity associated with
this protein. No adverse effects were observed in mice dosed with GOX
protein. Based on this study, in which the No-Observed-Effect-Level
(NOEL) for toxicity of the protein administered as a single dose was
considered to be 100 mg/kg (the highest tested target dose), no acute
dietary risks are posed for infants, children or adults.
3. Developmental/reproductive effects. No instances of reported
adverse reproductive or developmental effects to humans, domestic
animals or wildlife as a result of exposure to the GOX protein or the
microbial source of the gox gene, Achromobacter, are known. Enzyme
proteins have not been reported in literature to produce teratogenic
effects or reproductive deficiency when fed to animals or man (Pareza
and Foster, 1983).
4. Chronic effects. In an in-vitro digestive fate study, the GOX
protein was rapidly degraded in simulated gastric and intestinal fluid
with more than 90% of the initially added GOX protein degraded after 15
and 30 seconds incubation, respectively, as detected by western blot
analysis. Consequently, no chronic effects are expected for infants,
children or adults.
5. Carcinogenicity. Protein enzymes are not considered to be
carcinogenic (Pareza and Foster, 1983) and consequently, there is no
carcinogenic risk associated with the GOX protein for infants, children
or adults.
6. Endocrine effects. Not applicable. Enzyme proteins are not known
to interact or bind directly with the estrogen receptor to produce
endocrine effects. Further, there is little opportunity for systematic
absorption of the GOX protein due to rapid degradation by digestive
enzymes. Therefore, no adverse effects to the endocrine system is
known.
C. Aggregate Exposure
1. Dietary exposure. Oral exposure to the GOX protein at very low
levels may occur from ingestion of processed corn products; however,
the lack of mammalian toxicity and the digestibility of the protein
have been demonstrated as cited above.
2. Drinking water exposure. Transfer of the GOX protein to drinking
water is highly unlikely given containment of the protein in plant
cells and natural degradation upon plant senescence. Western blot
analysis has indicated that the GOX protein was not present in corn
pollen.
3. Non-occupational exposure. The GOX protein is produced at low
levels as a selectable marker and is contained within the cells of the
plant. Consequently, negligible exposure to the protein is expected
from handling corn seed, leaf or other plant tissue at planting, during
growth, or at harvest. In addition, negligible exposure to the GOX
protein would be expected during storage, transportation, or disposal
of insect-protected corn seed as the protein cannot drift or volatilize
from the plant and its bioactivity is rapidly lost upon decomposition
of the plant tissue.
D. Cumulative Risk
The GOX enzyme was isolated from an Achromobacter species and
catyalyzes the degradation of glyphosate to aminomethylphosphonic acid
(AMPA) and glyoxylate. This conversion of glyphosate to AMPA and
glyoxylate is the primary route for the degradation of glyphosate. This
degradation inactivates the herbicide and allows the plant cell
expressing the GOX protein to grow in the presence of glyphosate. This
mechanism is not shared by other known selectable markers used in
initial laboratory stages of plant cell selection. Consideration of a
common mode of toxicity is not appropriate given that there is no
indication of mammalian toxicity of the GOX protein and no information
that indicates that toxic effects would be cumulative with any other
compounds.
E. Safety Determinations
1. U.S. general population. The toxicity profile for the GOX
protein indicates no risk from exposure to the GOX protein by the
overall U.S. population. Monsanto believes that the lack of acute
toxicity, rapid digestibility of the GOX protein and lack of homology
to known proteinaceous allergens or toxins provide evidence for the
lack of toxicity and allergenicity and support an exemption from the
requirement of a tolerance for the GOX protein.
2. Infants and children. Monsanto considers the acute toxicity
data, the rapid degradation of the GOX protein in the mammalian
digestive system and the lack of homology to known proteinaceous
allergens as evidence to support the safety of this protein to infants
and children. Based upon this evidence, it is not expected that infants
and children would be more more susceptible to this protein than is the
adult population.
F. Residue Chemistry Data Summary
As a plant pesticide formulation inert ingredient, the gox gene was
used to produce the GOX protein which confers tolerance to glyphosate
as the selectable marker. The GOX protein is produced in plant tissues
including grain and forage at low levels as documented above. Mammalian
safety of the protein has been demonstrated in acute oral toxicity test
of the GOX protein. Analytical methods for the detection and
measurement of the GOX protein are not needed as Monsanto is
petitioning for an exemption from the requirement of a tolerance on the
basis of mammalian safety. The GOX protein is not on the Food and Drug
Administration's Generally Recognized as Safe (GRAS) list.
G. Environmental Fate Data Summary
The GOX gene was cloned from an Achromobacter species, reported to
be one of the most frequently occuring bacteria in the rhizosphere
(Joos et al., 1988). The GOX protein is produced at low levels within
the cells of the plant and expected to degrade at plant senescence and
exposure to physical, chemical and microbial processes in the
environment. (Mike Mendelsohn)
3. Seminis
PP 4E4310
A. Proposed Use Practices
Recommended application method and rate(s), frequency of
application, and timing of application. The inserted genes are under
the control of a constitutive promoter. Therefore, the viral coat
proteins will be produced within the tissues of the genetically
engineered plant and will not be applied externally. In information
provided to commercial growers, the resistance of the engineered plants
to specific plant viruses will be described. However, Seminis states
that no special instructions for use will be necessary. Appropriate
cultural practices for growing seed with genetically engineered virus
resistance will be determined by individual growers, as such practices
are for all other plant varieties.
B. Product Identity/Chemistry
1. Identity of the pesticide and corresponding residues. The
pesticide consists of a pair of viral coat proteins that are produced
by the genetically engineered plant. One protein consists of a fusion
of 16 amino acids of the cucumber mosaic virus coat protein and 281
amino acids of the watermelon mosaic virus 2 coat protein. The
molecular weight of the chimeric coat
[[Page 34276]]
protein is approximately 33,203. The second protein consists of 279
amino acids of the zucchini yellow mosaic virus coat protein with a
molecular weight of approximately 31,458.
2. Magnitude of residue anticipated at the time of harvest and
method used to determine the residue. The viral coat proteins are
expressed in plant tissues and, therefore, are not residues in the same
manner as a pesticide applied externally to growing crop plants.
Seminis believes that little concern exists for the presence of viral
coat proteins remaining on or in genetically engineered plants as they
are ubiquitous in nature, found in soil, water, terrestrial plants and
raw produce.
3. A statement of why an analytical method for detecting and
measuring the levels of the pesticide residue are not needed. The
Enzyme-Linked Immunoabsorbent Assay (ELISA) test can be used to
determine expression levels of viral coat proteins in transformed
plants, fruits and leaves. However, the available scientific literature
indicates that viral coat proteins do not pose a threat to human health
or the environment at any level. Therefore, Seminis states that an
analytical method for detecting and measuring the level of engineered
viral coat proteins is not needed.
C. Mammalian Toxicological Profile
Viral coat proteins are substances that viruses produce during a
plant infection to encapsulate and protect their genetic material. When
the genetic material encoding the coat protein for a plant virus is
introduced into a plant's genome, the plant is able to resist
subsequent infections by that same virus as well as strains closely
related to the donor virus. Virus-infected plants currently are and
have been a part of both the human and domestic animal food supply, and
Seminis agrees with EPA's finding that plant viruses are not known to
be harmful to humans (59 FR 60519-60535, November 23, 1994)(FRL-4755-
3). All available data from the scientific literature indicates that
plant viruses are not toxic to humans or other vertebrates.
Additionally, plant viruses are unable to replicate in mammals or other
vertebrates, eliminating the possibility of human infection. This has
been shown by injections of purified whole virus into laboratory
animals to develop antibodies for ELISA tests.
More importantly, however, this tolerance exemption will apply to
that portion of the viral genome coding for the whole coat protein and
any sub-component of the coat protein expressed in the plant. This coat
protein component alone is incapable of forming infectious particles.
Because whole intact plant viruses are not known to cause deleterious
human health effects, Seminis believes that it is reasonable to assume
that a subunit of these viruses likewise will not cause adverse human
health effects.
D. Aggregate Exposure
1. Dietary exposure. a. Food. Seminis states that the use of viral
coat protein-mediated resistance will not result in significant new
dietary exposure to plant viruses. Entire infectious particles of
zucchini yellow mosaic virus and watermelon mosaic virus, including the
coat protein component, are already found in the fruit and tissues of
many plants. Virus-infected food plants are and have been a part of the
human and domestic animal food supply. Such food plants and food
derived from them have been consumed, including by children and
infants, with no detectable or observed adverse effects to human
health.
b. Drinking water. Seminis states that the use of viral coat
protein-mediated resistance will not result in significant new levels
of viral coat proteins from engineered plants in drinking water. Plant
viruses are already present in soil and water. Viral coat proteins
expressed in genetically engineered plants are limited to plant
tissues. Upon plant senescence, viral coat proteins are believed to
degrade in the soil in the same manner as other proteins. Consequently,
Seminis believes that viral coat proteins produced as plant-pesticides
would represent a negligible addition to those existing in drinking
water.
2. Non-dietary exposure (lawn care, topical insect repellents,
etc.). The use of the genetically engineered viral coat proteins is for
improved virus disease resistance in agricultural crops. Therefore,
Seminis believes that non-dietary exposure to genetically engineered
viral coat proteins will be minimal to non-existent.
E. Cumulative Exposure
Exposure through other pesticides and substances with the common
mode of toxicity as this pesticide. Seminis believes that due to the
lack of toxicity associated with plant viruses and plant viral coat
proteins, cumulative effects with other pesticides and substances will
be non-existent.
F. Safety Determination
1. U.S. population. There is no known toxicity associated with coat
proteins from plant viruses. Consequently, a safety assessment is not
needed for these proteins. Given the long history of mammalian
consumption of the entire plant virus particle in foods, without any
adverse human health effects, Seminis reasonably believes that
consumption of a non-infectious component of the WMV plant virus is
safe. There are no known data that indicate aggregate exposure to plant
viral coat proteins under normal conditions will result in harm to any
person.
2. Infants and children. Viral coat proteins are ubiquitous in
foods, including those foods consumed by infants and children.
Moreover, there is not reason to believe that plant viral coat proteins
are likely to occur in different amounts in foods consumed by children
and infants. Further, there is no scientific evidence that viral coat
proteins used as plant pesticides would have a different effect on
children than on adults. Viral coat proteins are not toxic and,
therefore, Seminis believes with reasonable certainty that no harm will
result to Infants and Children from aggregate exposure to coat proteins
from plant viruses.
G. Existing Tolerances
An exemption from tolerance was granted for watermelon mosaic
virus-2 and zucchini yellow mosaic virus coat proteins as expressed in
Asgrow line ZW20 of Cucurbita pepo L. in November 1994 (59 FR 54824,
November 2, 1994)(FRL-4908-1).
H. International Tolerance
No known international tolerance or exemption from tolerance has
been granted for watermelon mosaic virus-2 and zucchini yellow mosaic
virus coat proteins. Seminis Vegetable Seeds, Inc. concludes that plant
viruses, including watermelon mosaic virus-2 and zucchini yellow mosaic
virus coat proteins, are not harmful to humans, and that there is a
reasonable certainty that no harm will result from aggregate exposure
to coat proteins of watermelon mosaic virus-2 and zucchini yellow
mosaic virus, and the genetic material necessary for production,
including all anticipated dietary exposures and all other non-
occupational exposures. Accordingly, Seminis believes that watermelon
mosaic virus-2 and zucchini yellow mosaic virus coat proteins qualify
for an exemption from the requirement of a tolerance in or on all raw
agricultural commodities. (Linda Hollis)
[FR Doc. 97-16659 Filed 6-24-97; 8:45 am]
BILLING CODE 6560-50-F
