United States       Prevention, Pesticides     EPA712-C-96-296
          Environmental Protection    and Toxic Substances     February 1996
          Agency         (7101)
&EPA    Microbialst Pesticide
           Test Guidelines
           OPPTS 885.1500
           Certification of Limits

-------
                           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."

-------
OPPTS 885.1500 Certification of limits.

     (a) Scope— (1) Applicability. This guideline is intended to meet test-
ing requirments of the Federal Insecticide, Fungicide, and Rodenticide Act
(FIFRA) (7 U.S.C. 163, et seq.\

     (2) Background. The source material used in  developing this har-
monized OPPTS  test guideline are OPP guidelines  151A-1  and 151A-
15.

     (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) Certification of ingredient limits.  (1) As required by  40 CFR
158.740, each registration must be supported by a certification of ingredi-
ent limits. Refer to OPPTS 830.1750 and 830.1800 regarding certification
of limits and analytical  methods to verify certified limits. The limits  for
MPCAs and contaminants should be expressed as:

     (i) MPCA units per unit weight or volume which may determined
using biological,  genetic, biochemical,  serological or other appropriate
tests.

     (ii) International units of potency per unit weight which may deter-
mined using  biological,  genetic, biochemical, serological or other appro-
priate tests.

     (iii) Weight percent of product.

     (2) Note that two  or more methods may  be required to verify  the
certified  limits of the MPCAs and any microbial impurities. Plate counts
(colony-  or plaque-forming units per unit weight or volume) or infectivity
assays may be necessary for quantitation and to allow preliminary identi-
fication. Antibiotic resistance bioassays may eliminate  some contamination
as well as provide additional identification support. In some cases,  espe-
cially in the  case of genetically  altered MPCAs, absolute identification
can  be achieved  only through the use  of one or more  of the various
immunological methods (such as enzyme-linked immunosorbent assay) or
molecular probe methods (such as  the  Southern  hybridization procedure

-------
or restriction endonuclease mapping). Methods must be specific enough
to determine the MPCA in the presence of revertants/mutants and contami-
nants that may have formed or been introduced during the replication/man-
ufacturing process.

-------