United States Environmental Protection Agency
Prevention, Pesticides and Toxic Substances (7101)
EPA 712-C-96-296   February 1996


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 
published 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. 
Environmental Protection Agency under the Toxic Substances Control Act 
(15 U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide 
Act (7 U.S.C. 136, et seq.).


     Final Guideline Release:  This guideline is available from the U.S. Government Printing Office, Washington, DC 20402 on The Federal Bulletin Board. By modem dial 202ÿ09512ÿ091387, telnet and ftp: fedbbs.access.gpo.gov (IP 162.140.64.19), internet: http://fedbbs.access.gpo.gov, or call 202ÿ09512ÿ090132 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 testing 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 
harmonized 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 information 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 
ingredient 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 
determined using biological, genetic, biochemical, serological or other 
appropriate 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 identification. Antibiotic resistance bioassays may 
eliminate some contamination as well as provide additional 
identification support. In some cases, especially 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 contaminants that may 
have formed or been introduced during the replication/manufacturing 
process.