United States       Prevention, Pesticides     EPA712-C-96-294
          Environmental Protection    and Toxic Substances     February 1996
          Agency         (7101)
&EPA    Microbial Pesticide
          Test Guidelines
          OPPTS 885.1300
          Discussion of Formation
          of Unintentional
           ngredients

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                           INTRODUCTION
     This guideline is one  of a  series  of test  guidelines that have been
developed by the Office of Prevention, Pesticides and Toxic Substances,
United States Environmental  Protection Agency for use  in the testing of
pesticides and toxic substances, and the  development of test data that must
be submitted to the Agency  for review under Federal regulations.

     The Office of Prevention, Pesticides and Toxic Substances (OPPTS)
has  developed this guideline through  a process of harmonization that
blended the testing  guidance  and requirements that  existed in the Office
of Pollution Prevention and  Toxics  (OPPT) and appeared in Title  40,
Chapter I,  Subchapter R of the Code of Federal Regulations  (CFR),  the
Office of Pesticide Programs (OPP) which appeared in publications of the
National Technical  Information Service (NTIS) and the guidelines pub-
lished by the Organization  for Economic Cooperation and Development
(OECD).

     The purpose of harmonizing these  guidelines  into a single set of
OPPTS guidelines is to minimize  variations among the testing procedures
that must be performed to meet the data  requirements of the U. S. Environ-
mental Protection Agency  under  the Toxic  Substances  Control Act  (15
U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide Act
(7U.S.C. I36,etseq.).

     Final  Guideline Release: This guideline  is available from the U.S.
Government Printing Office, Washington, DC 20402 on The Federal Bul-
letin   Board.   By  modem  dial   202-512-1387,  telnet   and   ftp:
fedbbs.access.gpo.gov    (IP     162.140.64.19),    internet:     http://
fedbbs.access.gpo.gov, or call 202-512-0132 for disks  or paper copies.
This guideline is also available electronically in ASCII and PDF (portable
document format) from the EPA Public Access Gopher  (gopher.epa.gov)
under the heading "Environmental Test  Methods and Guidelines."

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OPPTS 0885.1300 Discussion of formation of unintentional ingredi-
ents.
     (a) Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of the  Federal Insecticide,  Fungicide, and  Rodenticide
Act (FIFRA) (7 U.S.C. 136, et seq.}.

     (2) Background. The source material used in developing this har-
monized OPPTS test guideline are OPP guidelines 151A-1  and  151A-
12.

     (b) Information needed. The product analysis data requirements for
microbial pest  control agents (MPCAs) parallel those for conventional
chemical pesticides in OPPTS Series  830. However,  due to  the  unique
nature, composition, and mode of action of the MPCAs,  there are some
important differences. For example, protozoa, bacteria, fungi, and viruses
should be identified to the extent possible by taxonomic position, serotype,
composition, and strain, or by any other appropriate specific means. This
information would take the place of chemical name and structure informa-
tion  for conventional chemical pesticides. In addition, the Agency must
be reasonably assured that the methods used and the  data submitted are
capable of demonstrating that the microbial pesticide used in  the field is
the same as that which was tested for safety.

     (c) Registration applications. Each registration application shall in-
clude the following information:

     (1) Theoretical discussion. The theoretical discussion concerns the
formation of each substance, aside from the control agents  and  inten-
tionally added,  chemically characterized active and inert ingredients, that
might reasonably be present in the pesticide product, as outlined OPPTS
830.1670. Examples of such extraneous materials are: Allergens, microbial
toxins, and other metabolic products; mutant strains; microbial contami-
nants with  particular reference to potentially infective  or  antagonistic
forms; side products from chemical reactions employed in the manufactur-
ing process, fermentation residues from the growth of bacteria or fungi;
extraneous host residues from viruses produced in cell cultures, whole ani-
mals, or other living forms; residues of contaminants that remain following
the purification  or extraction  process; and impurities  in  chemicals  used
in the manufacturing process. The  discussion  shall include the procedures
used to ensure the purity of the unformulated MPCA preparation; if purity
(within reasonable limits) cannot be achieved, then  the means of control-
ling contaminant levels to an acceptable limit must be delineated.

     (2) Toxic or sensitizing substances. If substances toxic or sensitizing
to humans or other nontarget mammalian species are known or suspected
to be present at any stage of the manufacturing process,  then data must
be submitted to show that the substances do not exist in the final product
or exist only in  quantities too small to pose any hazard.

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     (3) Human or animal pathogens. Human or other nontarget animal
pathogens such as (but not  limited to) Shigella,  Salmonella, and  Vibrio
must not be present at hazardous levels in the technical grade of the active
ingredient. If the production method can support the growth of human or
animal pathogens, each production batch must be tested for their presence.
Each application for registration of a manufacturing-use product or end-
use product should contain an analysis of all human or animal pathogens
that might be present at potentially hazardous levels in the product before
formulation. The  application should propose methodology for detecting
and/or eliminating these from the product. For  example,  the method pre-
scribed in 40 CFR 180.1011 to  monitor each production batch of Bacillus
thuringiensis  for the pathogen  Bacillus anthracis involves subcutaneous
injection of at least  1 million viable microorganisms or  spores into each
of 5 laboratory mice weighing  17 to 23 g. Such tests shall show no evi-
dence of infection or injury in the test animals when observed for 7 days
following injection.

     (4) Detection of hazardous contamination.  Methods should be pro-
posed for controlling any hazardous  contamination detected in the product.
Discarding of a contaminated production batch or treating the formulation
may be  equally  acceptable  methods for controlling the contaminating
microorganisms.

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