[Federal Register Volume 63, Number 67 (Wednesday, April 8, 1998)]
[Notices]
[Pages 17174-17176]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 98-8659]


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ENVIRONMENTAL PROTECTION AGENCY

[PF-800;FRL-5781-1]


Notice of Filing of Pesticide Petition

AGENCY: Environmental Protection Agency (EPA).
ACTION: Notice.

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SUMMARY: This notice announces the initial filing of petition (PP 
7F4822), submitted by Monsanto Company, proposing the establishment of 
a regulation for an exemption from the requirement of a tolerance for 
residues of the plant pesticide, active ingredient, Bacillus 
thuringiensis variety kurstaki (B.t.k.) insect control protein 
(CryIIA), when used in or on all food and feed crops.

DATES: Comments, identified by the docket control number PF-800, must 
be received on or before May 8, 1998.
ADDRESSEES: By mail submit written comments to: Public Information and 
Records Integrity Branch (7502C), Information Resources and Services 
Division, Office of Pesticides Programs, Environmental Protection 
Agency, 401 M St., SW., Washington, DC 20460, In person bring comments 
to: Rm. 119, CM #2, 1921 Jefferson Davis Highway, Arlington, VA, 22202.
    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 of the 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. 119 at the 
address given above, from 8:30 a.m. to 4 p.m., Monday through Friday, 
excluding legal holidays.

FOR FURTHER INFORMATION CONTACT: By mail: Willie H. Nelson, 
Biopesticides and Pollution Prevention Division (7511W), Office of 
Pesticides Programs, Environmental Protection Agency, 2800 Crystal 
Drive, Arlington, VA 22202, (703) 308-8682; e-
mail:[email protected].

SUPPLEMENTARY INFORMATION: EPA has received petitions as follows 
proposing the establishment and/or amendment of regulations for 
residues of certain pesticide chemicals/microbials in or on various 
food commodities under section 408 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 
petitions.
    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-800] (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 included any information claimed as CBI, is available for 
inspection from 8:30 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:
[email protected]



[[Page 17175]]


    Electronic comments must be submitted as an ASCII file avoiding the 
use of special characters and any form of encryption. Comments and data 
will also be accepted on disks in Wordperfect 5.1/6.1 file format or 
ASCII file format. All comments and data in electronic form must be 
identified by the docket number [PF-800] and appropriate petition 
number. Electronic comments on this 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: March 20, 1998

Janet L. Andersen,
Director, Biopesticides and Pollution Prevention Division, Office of 
Pesticides Programs.

Summary of Petition

    Below a summary of the pesticide petition is printed. The summary 
of the petition was prepared by the petitioner. This 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.

Monsanto Company

PP 7F4822

    1. Plant-pesticide uses. Cotton, Gossypium hirsutum, has been 
genetically engineered to be resistant to selected insect pests of the 
taxonomic order Lepidoptera. Insect protection was accomplished by the 
insertion of the cryIIA gene from Bacillus thuringiensis subsp. 
kurstaki (B.t.k.) which encodes for the production of a protein 
specifically insecticidal to Lepidopteran larvae in cotton but safe to 
nontarget organisms such as mammals, birds, fish and beneficial 
insects. Larvae of Lepidopteran pests are the most important insect 
pests impacting successful cotton production and numerous chemical 
insecticide treatments are typically applied for their control. The 
production of cotton varieties containing the CryIIA gene from B.t.k. 
is expected to significantly reduce chemical insecticide use in cotton 
and; therefore, provide a major benefit to cotton growers and the 
environment.
    2. Safety. The CryIIA protein produced in BollgardTM 
Cotton is >99.9% identical to the protein produced by the B.t.k. HD-1 
bacterial strain found in nature and in commercial B.t.k. formulations 
registered with the EPA. These microbial B.t.k. formulations have been 
commercially available for the last 30 years. This strain controls 
insect pests by the production of crystalline insecticidal proteins 
known as delta-endotoxins. To be active against the target insect, the 
protein must be ingested. In the insect gut, the protein binds to 
specific receptors on the insect mid-gut, inserts into the membrane and 
forms ion-specific pores. These events disrupt the digestive processes 
and cause the death of the insect.
    There are no receptors for the protein delta-endotoxins of B. 
thuringiensis subspecies on the surface of mammalian intestinal cells; 
therefore, humans are not susceptible to these proteins. This has been 
confirmed in numerous safety studies carried out in laboratory animals 
which are traditionally experimental surrogates for humans. The results 
of some of these studies have been published in scientific reviews 
(Ignoffo,1973; Shadduck et al., 1983; Siegel and Shadduck, 1990). 
Results of unpublished safety studies generated by registrants of B. 
thuringiensis commercial preparations have also been summarized in a 
recently issued EPA Registration Standard for Bt Formulations (EPA, 
1988). In published reviews and the EPA document, studies are 
referenced in which large doses (5,000 mg/kg) of B. thuringiensis 
formulations were administered as single or multiple oral doses (up to 
2 years) to different laboratory animals, with no adverse effects.
    Avian and aquatic organisms have also been fed B. thuringiensis 
formulations, with no adverse effects. A typical formulation is 
composed of Bt spores and Bt protein endotoxin, the latter compromising 
up to one-third of the weight of the spores. While target insects are 
susceptible to oral doses of B.t.k. proteins, there was no evidence of 
any toxic effects observed in non-target laboratory mammals, fish or 
birds given the equivalent of up to 106 g of protein per 
gram of body weight. No deleterious effects were observed on non-target 
insects at doses over 100 fold higher than needed to control target 
insects (EPA 1988).
    In addition to the lack of receptors for the B.t.k. proteins, the 
absence of adverse effects in non-target animals is further supported 
by the poor solubility and stability of the B.t.k. proteins in the acid 
milieu of the stomach. The acid conditions in the stomach and the 
presence of bile acids denature the B.t.k. proteins facilitating their 
rapid degradation by pepsin. In vitro enzymatically activated delta-
endotoxins are also non-toxic when administered orally to laboratory 
animals (Nishitsutsuji-Uwo et al. 1980). Even if activated B.t.k. 
protein toxins could enter the mammalian gastrointestinal tract, there 
are no receptors on the surface of gastrointestinal tissues to permit 
binding of the protein toxin to the cell surface. These scientific 
considerations are experientially support by the history of completely 
safe use of B. thuringiensis preparations. Based on the available 
scientific data, EPA and other regulatory scientists worldwide have 
determined that use of registered B. thuringiensis products pose no 
risks to human health or non-target organisms.
    Monsanto Company has also submitted several toxicology studies in 
support of the CryIIA protein as a plant pesticide. According to 
Monsanto Company, there is no acute toxicity of the CryIIA protein. In 
addition, the CryIIA protein is also produced at low levels by Bollgard 
cotton plants and is contained within the cells of the cotton plant. 
Consequently, there would be negligible exposure to the protein from 
handling cottonseed, leaf tissue or lint at planting, during growth, or 
at harvest. In addition, there would be no potential hazard during 
storage, transportation, or disposal of Bollgard cottonseed as the 
protein cannot drift or volatilize from the plant and its bioactivity 
is rapidly lost upon decomposition of the plant tissue.
    The following mammalian toxicity studies have been conducted to 
support this exemption from the requirement of a tolerance:
    i. A mouse acute oral gavage study in which the No-Observed-Effect-
Level (NOEL) for toxicity of the CryIIA protein administered as a 
single dose was considered to be 4,000 mg/kg (the highest tested dose).
    ii. In vitro digestive fate of the CryIIA protein in simulated 
gastric and intestinal fluids. The results of this study established 
that the CryIIA protein and its associated functional activity will be 
efficiently degraded upon exposure to gastric and intestinal fluids in 
the mammalian digestive tract. A lack of stability to digestion is a 
characteristic of proteins which are non-allergens.
    iii. Amino acid sequence homology assessment of the CryIIA protein 
to known allergens and toxins. The results of this analysis establish 
that the CryIIA protein expressed in Bollgard cotton shares no 
significant sequence similarity with known toxins, allergens or gliadin 
proteins. In addition, the CryIIA protein

[[Page 17176]]

appears to contain no sequences relevant to allergy or coeliac disease.
    3. Threshold effects-- i. Acute toxicity. Based on the available 
acute toxicity data for the CryIIA protein and on the safe use of 
microbial Bacillus thuringiensis foliar formulations containing the 
same protein and registered with the EPA and used commercially for 30 
years, no acute dietary risks are posed.
    ii. Chronic effects. The CryIIA protein is degraded upon exposure 
to gastric and intestinal fluids in the mammalian digestive tract. 
Consequently, no chronic effects are expected. In addition, in 
published reviews and the EPA Registration Standard for Bt Formulations 
(EPA, 1988) studies are referenced where large doses (5,000 mg/kg) of 
B. thuringiensis formulations were administered as single or multiple 
oral doses (up to 2 years) to different laboratory animals, with no 
adverse effects.
    4. Non-threshold effects. Carcinogenicity: Proteins are not 
considered to be carcinogenic (Pareza and Foster, 1983) and 
consequently, there is no carcinogenic risk associated with the CryIIA 
protein.
    5. Aggregate exposure. Cottonseed meal is not currently used for 
human consumption in the United States (Morgan, 1990; Cottonseed Oil, 
1990). The presence of gossypol and cyclopropenoid fatty acids in 
cottonseed also limits its use as a protein supplement in animal feed 
except for cattle, which are unaffected by these components. 
Inactivation or removal of these components during processing, which 
entails heating and chemical treatment, enables the use of some 
cottonseed meal for catfish, poultry and swine. However, as the CryIIA 
protein is heat labile, the biological activity of the protein is 
expected to be lost upon processing as demonstrated by Sims and 
Berberich with other B.t. proteins (1996).
    Refined cottonseed oil and cottonseed linters (the fiber remaining 
after ginning seed cotton) are also highly processed and are the only 
cotton products consumed as food by humans. Cottonseed oil is typically 
removed from the meal by direct solvent extraction with hexane and is 
further processed and refined by exposure to extreme heat and alkaline 
pH. Processed cottonseed oil contains no detectable protein (Fuchs, 
1994; Fuchs et. al., 1993). Cotton linters are essentially comprised 
only of cellulose (>99.9%) and Sims et. al. (1996) have demonstrated 
that processed linters, which also undergo exposure to temperatures 
exceeding 100 deg.C and alkaline treatment do not contain detectable 
levels of transgenic proteins such as CryIIA.
    Based on these results, aggregate exposure to the CryIIA protein 
through ingestion of cottonseed oil and linters derived from bollgard 
cotton would be negligible.
    6. Determination of safety for U.S. population. The toxicity data 
support an exemption from the requirement of a tolerance for the CryIIA 
protein expressed in Bollgard cotton indicate that there would be no 
risk from exposure to the CryIIA protein by the overall U.S. 
population. In addition, the CryIIA protein expressed in Bollgard is 
more than 99.9% identical to the natural protein, which is component of 
microbial Bacillus thuringiensis subsp. kurstaki formulations that have 
been registered with the EPA and available commercially for the last 30 
years. The EPA and other regulatory scientists worldwide have 
determined that use of registered B. thuringiensis products pose no 
significant risks to human health or non-target organisms (EPA, 1988).
    7. Determination of safety for infants and children. Monsanto 
considers the acute toxicity data, the rapid degradation of the CryIIA 
protein in the mammalian digestive system, the lack of homology to 
known proteinaceous allergens or toxins and a 30 year history of safe 
use of microbial B. thuringiensis  containing the near identical CryIIA 
protein as ample evidence to support the safety of this protein to 
neonatal infants, infants and children.
    8. Estrogenic effects Not applicable. Proteins are not capable of 
direct estrogenic activity as they are incapable of binding to an 
estrogen receptor.
    9. Chemical residue. Not applicable. In the United States, only 
refined cottonseed oil and cottonseed linters (the fiber remaining 
after ginning seed cotton), which are highly processed, are the only 
cotton products consumed as food by humans. Cottonseed oil is typically 
removed from the meal by direct solvent extraction with hexane and is 
further processed and refined by exposure to extreme heat and alkaline 
pH. Processed cottonseed oil contains no detectable protein (Fuchs, 
1994; Fuchs et. al., 1993). Cotton linters are essentially comprised 
only of cellulose (>99.9%) and Sims et. al. (1996) have demonstrated 
that processed linters, which also undergo exposure to temperatures 
exceeding 100 deg.C and alkaline treatment do not contain detectable 
levels of transgenic proteins such as CryIIA.
    10. Environmental fate. The CryIIA protein expressed in Bollgard 
cotton plant tissue was evaluated over 120 d in both a laboratory 
microcosm and under field conditions. DT50 values were 15.5 
d and 31.7 d for the laboratory and field respectively. These results 
demonstrate that CryIIA protein, as a component of post-harvest 
Bollgard cotton plants, will dissipate when cultivated into soil.

Literature Cited

    1. Cottonseed Oil. 1990. eds. L.A. Jones and C.C. King. National 
Cottonseed Products Association, Inc. and The Cotton Foundation, 
Memphis.
    2. EPA, 1988. Guidance for the Reregistration of Pesticide Products 
Containing Bacillus thuringiensis as the Active Ingredient. NTIS PB 89-
164198.
    3. Fuchs, R.L., Berberich, S.A. and Serdy, F.S. 1993. Safety 
evaluation of genetically engineered plants and plant products: insect-
resistant cotton. In ``Biotechnology and Safety Assessment.'' J.A. 
Thomas and L.A. Myers, eds. Raven Press Ltd., New York, pp. 199-212.
    4. Fuchs, R.L., 1994. Gene Expression and Compositional Analysis 
from Field-Grown Insect Resistant Cotton Tissues, Study Number 92-01-
36-07, an unpublished study conducted by Monsanto Company. EPA MRID# 
43168701
    5. Ignoffo, C.M. 1973. Effects of Entomopathogens on Vertebrates. 
Ann. N.Y. Acad. Sci. 217:144-172.
    6. Morgan, S.E. 1990. Gossypol Residues in Organ Meats vs 
Thresholds of Toxicity. Vet. Hum. Toxicol. 32S:76.
    7. Nishitsutsuji-Uwo and Yasuhisa Endo et. al. 1980. Mode of Action 
of Bacillus thuringiensis -Endotoxin: Effect on TN-368 Cells. Appl. 
Ent. Zool. 15:133-139.
    8. Pareza, M.W. and Foster, E.M. 1983. Determining the stability of 
enzymes used in food processing. J. Food. Prot. 46:453-468.
    9. Siegel and Shadduck, 1989, Safety of Microbial Insecticides to 
Vertebrates and Humans, In Safety of Microbial Insecticides. CRC Press, 
Inc., FL. pp 101-113.
    10. Sims, S.R. and Berberich, S.A. 1996. Bacillus thuringiensis 
CryIA protein levels in raw and processed cottonseed of transgenic 
cotton: determination using insect bioassay and ELISA. J. Econ. 
Entomol. 89:247-251.
    11. Sims, S.R., Berberich, S.A., Nida, D.L., Segalini, L.L., Leach, 
J.N., Ebert, C.C. and Fuchs, R.L. 1996. Analysis of expressed proteins 
in fiber fractions from insect-protected and glyphosate-tolerant cotton 
varieties. Crop Physiol. Metabol.36:1212-1216.

[FR Doc. 98-8659 FIled 4-7-98; 8:45 a.m.]
BILLING CODE 6560-50-F