Guidelines CIB RC

t
F.N o. 19 -65 | 2022-CIR-I

qt-{f,g{fr-R
Government of India
g.R\nimqno-qurriTff,q
Ministry of Agriculture & Farmers Welfare
pB\i?ifrr{rlo-ilqftrrFI
Department of Agriculture & Farmers Welfare
qqffi ftqfur, €rn]q f:.i sq6 fraqT-dq
DIRECTORATE OF PLANT PROTECTION, QUARANTINE & STORAGE
+-flq offi frd qqq q-fi-*-{q qfrfr
Central Insecticides Board and Registration Committee
\r{. qq. +, q-ftqrdlE GftqTUD_l21001
N.H. IV. FARTDABAD (HARYANA)-I21001
******
Dated: o {\n"r" ber, 2022
PUBLIC NOTICE
Subject: Guidelines ldata requirement for grant of registration under various categories -
reg.
The Registrarion Committee in its 442"r meeting held on 18.11.2022 has confirmed the
guidelines approved in 440th RC meeting (Agenda item no. 10.58) with some minor coruections.
ihe finalized guidelines/data requirement for grant of registration under various categories vide
minutes of its 442"d meeting (Agenda Item No. 1.0) are enclosed for information of all
stakeholders. These guidelines shall be applicable from 03.08.2022 as per the decision of RC in
its 440tr'RC meeting.
This has the approval of Secretary (CIts&RC).
__2
Encl.: Asabove.
W
-\Govin'd Ram)
--;--
Sr. Administrative Officer
Copy to:
1. Chairman, Registration Comrnittee
2. PPS to JS (PP)/ PPS to PPA
3. IT Cell, HQ, Faridabad for uploading the same on the website.
Minutes of 442nd RC meeting held on 18.11.2022
Annexure 1.0.1
Guidelines/Data requirement for grant of registration under various categories:
1. GUIDANCE TO STAKEHOLDERS:
A. Stakeholders generating data and submitting the application for registration should ensure that the
tests are conducted and data generated in accordance with established scientific procedures
following the test guidelines and the principles of Good Laboratory Practices. The data should be
authentic, replicable, utilizable and of good quality. The complete study reports should be
submitted.
B. The requirement for registration usually includes data and information on proposed application; data
on identity of the insecticide (identity, composition, analysis and quality); data to assess risk to
humans and the environment; data to assess efficacy of the product; and the packaging and labeling
requirements.
C. The data requirement for registration of insecticides varies with the type of insecticides to be
registered (i.e. chemical or bio-pesticide and also type of bio-pesticide i.e. Microbial Pest Control
Agent (MCPA) or Botanical or Semiochemical/Pheromone); the type of material to be registered i.e.
Technical or Formulation or Manufacturing Use Product (MUP); the type of formulation; solid (WP,
granules, powder) etc. liquid (EC, EW, SC etc.) or Vapor (vaporizer, fumigants etc.); the category of
registration – i.e. provisional [u/s 9(3B)]; regular [u/s 9(3)] or subsequent “Me-too” [u/s 9(4)] as per
the provisions (Please refer section (9) of the Insecticides Act, 1968); and purpose of registration –
domestic use or export or for both (domestic use and export); the intended use of the pesticide to be
registered or its label claims etc. Hence, before starting data generation or submitting application for
registration, the applicant should ensure that the requirements are being complied correctly for the
type of pesticide to be registered under the desired category and for the intended purpose.
D. The data submitted by the applicant at the time of seeking registration under section 9(3b)
shall not be required to resubmit again at the time of submission of application for regular
registration under section 9(3) by the same applicant, if chemical composition and other claims
remain unchanged.
2. GUIDELINES FOR DATA REQUIREMENT FOR REGISTRATION OF PESTICIDES
These guidelines provide the guidance to the stakeholders to generate data and submit application with the
indicated requirements. The data requirements are divided into five segments, as envisaged below:
(A) Legal Requirements: The documents required from legal discipline are at Annexure I.
(B) The Data Requirements for Chemical Pesticides are at Annexure II.
(C) The Data Requirements for Microbial Pest Control Agents (MPCA) Pesticides are at Annexure III.
(D) The Data Requirements for Botanical/ Plant origin Pesticides are at Annexure IV.
(E) The Data Requirements for Pheromones/Semio-chemicals are at Annexure V.
Note:
1. THE REQUIREMENTS/GUIDELINES FOR REGISTRATION OF INSECTICIDES

EXCLUSIVELY FOR EXPORT REMAIN THE SAME AS APPROVED/RECOMMENDED
BY THE REGISTRATION COMMITTEE IN ITS 380TH RC AND ARE AVAILABLE ON THE
WEBSITE.
2. The abbreviations used in the guidelines are in APPENDIX- I
3. The terms used in the report are defined in APPENDIX-II
LEGAL REQUIREMENTS
ANNEXURE-I
A. Legal Requirements:
1. Form – I duly signed/digitally signed
2. Copy of BOD Resolution/Affidavit/Partnership deed (Notarized)
3. Affidavit for Chemical composition on NJSP (Notarized )
4. Certificate as per category of Industry/ Manufacturing license (Notarized)
5. PAN No. (Notarized)
6. Incorporation Certificate (Notarized) and other documents as per KYC requirement
7. Proof of ownership/lease agreement of manufacturing site purported to be used as manufacturing site

(Notarized)
8. Technical to be used in formulation should be duly registered (In case of import - Reference of RC
meeting in which it was approved). {Only deemed registration status without issuance of Certificate of
registration shall not be considered}.
9. Name of registrant of Formulation Import (Reference of Registration Committee Meeting in which

formulation import was approved).
10. Letter of consent, duly legalized from Indian Embassy/High Commission/ Consulate/ apostle
documents in the Country of origin /(Applicable in case of 9(4) TI/FI Applications).
11. List of products for which the registration has been given to the firm & Manufacturing License
obtained and products actually manufactured during the previous 3 (three) years (Applicable in case
of 9(4) TI/FI & FIM Applications.
Note: The RC has decided to implement the KYC as per decision taken in its 424 th meeting. As per
requirement the module has been brought to working condition. The applicant are supposed to feed
their documents are per requirement.
If, the “KYC” is activated, then the applicant needs not to submit the following documents
repeatedly:
1. Copy of BOD Resolution/Affidavit/Partnership deed (Notarized)
2. Certificate as per category of Industry/ Manufacturing license (Notarized)
3. PAN No. (Notarized)
4. Incorporation Certificate (Notarized) and other documents as per KYC requirement
5. Proof of ownership/lease agreement of manufacturing site purported to be used as manufacturing
site(Notarized)
6. List of products for which the registration has been given to the firm and Manufacturing License
obtained and products actually manufactured during the previous 3 (three) years.
.
ANNEXURE-II
DATA REQUIREMENTS FOR CHEMICAL PESTICIDES
Data Requirements for Chemical Pesticides:
A. Chemistry:
9(3B) 9(3) 9(4)
Sl.
Parameter
No. TI TI FIM TI TIM FI FIM TI TIM FI FIM
M
1 2 3 4 5 6 7 8 9 10 11 12 13
A. CHEMISTRY
1. Details of source of R NR R R NR R R R NR R R
supply of Technical
2. Chemical Composition R R R R R R R R R R R
(clearly showing claims
of purity of active
ingredient, impurities or
adjuvants, as the case
may be) in Form-I and
L/L
3. Physical and Chemical R R R R R R R NR R NR NR

Properties of the active
ingredient in Technical
and adjuvant in case of a
formulation
4. Technical Bulletin R NR NR R NR R NR NR NR NR NR
5. Product Specification in R R R R R R R NR R* NR NR
BIS format/BIS No. if
published
6. Method of Analysis R R R R R R R NR R* NR NR
7. Analytical Test Report R R R R R R R NR R NR NR

(ATR) from GLP/NABL
accredited laboratory
8. Characterization (Identity R R NR R R NR NR NR R NR NR
Test) of active ingredient
by UV-VIS, IR, MS and
NMR spectra)
9. Identification& R R NR R R NR NR NR R NR NR
Quantification of
Impurities
10. Shelf-life claim R R R R R R R R R R R
11. Storage Stability Data R R R R R R R NR NR NR NR

(samples stored in three
varied agro-climatic
conditions) for six
months in excess of
claimed shelf-life along
with meteorological data
for corresponding period
12. Establishment of NR NR NR NR R*/ R*/ R*/ R R R R

Chemical Equivalence
NR* NR* NR*
13. Detailed stepwise R R R R R R R NR R NR NR

manufacturing process
(provide chemical
reactions explained with
structural formulae, all
applicable reaction other
conditions in case of
technical grade
insecticides including
flow sheet diagram).
14. Information about Raw R R R R R R R NR R NR NR

Materials Used along
with their source of
supply
15. Effluent Treatment R R R R R R R NR R NR NR

method with complete
details
16. Legalized letter of R NR R R NR R R R NR R NR

consent that the
manufacturer is registered
for the Technical Grade
Pesticide/ Insecticide and
that he consents
supplying the Technical
Grade Pesticide/
Insecticide to the
applicant.
17. Registration Certificate R NR NR R NR R NR R NR R NR

from DNA of the
pesticides from source
country
18. In case the supplies are to R NR NR R NR R NR R NR R NR

be made through a
supplier, a duly legalized
certificate from the
exporting manufacturer
that the supplier is his
authorized agent and that
the invoice would

originate from the
approved source of
import (actual
manufacturer).
19. An In-Process sample to R R R R R R R NR R NR NR

be drawn from the
R&D/manufacturing
facility of applicant in
case of technical
indigenous manufacture
of the insecticide/sample
to be submitted in case of
technical import$; along
with certified reference
material (CRM) and
standard impurities
alongwith purity
certificate in case of
technical grade
insecticides for pre-
registration verification in
the Central Insecticides
Laboratory, Faridabad
(CIL). In case of FIM
category sample and
CRM to be submitted in
CIL Faridabad
20. Methodology for residue R R R R R R R NR NR NR NR

estimation in BIS format.
Remarks on chemistry parameters:
1. S. No. (6) & (7) R* wherever If BIS published; than not required, if not, published Registered product
specification is required.
2. S. No. 12. Not required (NR)- For first Registration, Required (R)- for subsequent registration after the first
registration.
3. Adjuvant(s) shall be mentioned by their common names(s) and not by code names or numbers and their
complete chemical identity shall be provided. No patented adjuvant shall be incorporated in the claimed
chemical composition.
4. Sample shall be drawn in case of 9(3) TI, TIM and 9(4) TIM as per approved procedure in 345 th RC or as
and when amended by RC , a proposal may be sent to DA&FW for their concurrence of drawl of In-Process
sample from the technical manufacturing site of importer, till then submission of sample in CIL in case of TI
shall be permitted.
5. In case of the insecticides for Seed Treatment, ‘Adhesion to Seed Test’ shall be invariably provided.
6. Same method of analysis should be used in generating ATR and shelf-life data.
7. Accelerated Storage Data can be considered for grant of provisional shelf-life. However, in such cases the
Certificate of Registration (CR) shall be issued with a validity of two years. Shelf-life claim of up to 2-years
or as the case may be (provisionally) be granted to the insecticides with a condition that applicant is required
to submit real time / actual storage stability study data in the proposed construct and container of sale for
duration of minimum 30 months, within two and half years of submission of application for granting the
registration, failing which Registration Certificate shall stand invalid.
8. Data requirement for registration of LongLasting Insecticide Impregnated/ Incorporated Mosquito Bed Nets
shall be for 9(3b) and 9(3) category only.
9. Data requirement for registration of Petroleum derived products like spray oilnatural mineral oil products
shall be for 9(3b) and 9(3) category only.
10. Data at S. No. 7-9 shall not be older than 5 years on the date of application in case of TI.
11. In case of FI-WRT (Formulation Import without registering Technical) OR FIM-WRT (Formulation
Indigenous Manufacture without registering Technical), in addition to data on formulation, complete
chemistry data on technical including shelf life and packaging data of technical as per the guidelines of
TI/TIM should also be submitted along with chemical composition on Rs 10/- Non-Judicial Stamp Paper
(NJSP). The applicant of FI-WRT are encouraged to seek registration for Technical also.
12. In case of 9(3b) TI, 9(3) TI/FI & 9(4) TI/FI category, a condition in CR to be incorporated as an
analytical test report by the manufacturer (exporting to India) about the quality of the insecticide/Pesticide
from a NABL or ISO 17025:2017 compliance /GLP certified laboratory. (Such analytical test report in
respect of the batch(s) shall accompany each & every consignment exported to India)
$
13. It was decided that the in-process sample also be drawn in case of TI/FI-WRT. A proposal shall be
submitted to DA&FW for in-principle approval. Till such approval is obtained existing policy should be
continued.
14. In case of formulation of pre-mix combination product having three active ingredients, a single method
of analysis must be used.
B. Bio-efficacy:(Insecticide/Fungicide/Herbicide/PGR):
Table-1
Sr. Parameters 9(3b) 9 (3) 9(4)

No.
TI TIM FIM TI TIM FI FIM Label TI/TIM/FI/F
expans IM or
ion endorsement
of already
approved
label
expansion.
Bio-efficacy
1. Bio-effectiveness R$ R$ R* R$ R$ R R** R** No data
**
requirement.
2. Phytotoxicity R$ R$ R* R$ R$ R R** R** The claim will
**
be granted as
$ $ * $ $
3. Effect on germination of R R R R R R R** R** per approved
** formulation
Seed (in case of seed
treatment of fungicide u/s 9(3)/
and insecticide) approval
4. Effect on parasitoids and N NR R* NR NR R R** R** claims of
** label
predators (Insecticides& R
PGR) expansion.
5. Effect on beneficial soil N NR R* NR NR R* R** R**
*
micro-organisms & R
physico-chemical
properties (Herbicide
only).
Effect on beneficial soil
micro-organisms in case
of Seed treatment and on
soil applied pesticides
(Fungicides, Insecticide
and PGRs).
6. Translocation in Plants R R NR R R N NR NR
R
7. Metabolism in Soil R R NR R R N NR NR
R
8. Metabolism in Water R R NR R R N NR NR
R
9. Metabolism in Plant R R NR R R N NR NR
R
10. Persistence in Soil R R R R R R R NR
11. Persistence in Water R R R R R R R NR
12. Persistence in Plant R R R R R R R R
13. Compatibility with N NR R NR NR R R R
other chemicals, if R
claimed
14. Residues in Plant N NR R# NR NR R# R# R
R
15. Residues in Soil N NR R# NR NR R# R# NR
R
16. Residue tolerance limits N NR R NR NR R R R
fixed by foreign R
countries
17. Cost Benefit Ratio/ Per N NR NR NR NR R R R
Rupee return (per ha) R
18. Registration status in N NR R R NR R R R
foreign countries R
19. MRL Performa along N NR R NR NR R R R
with Pen drive in R
duplicate (other than seed
treatment).
20. Label and Leaflets (as R R R R R R R R
per Insecticides Rule 18
and 19)
Remarks on Bioefficacy parameters:
a. At S. No. 5- Applicant may submit position paper and published data related to toxicological effects
on beneficial insects in case FI/FIM.
b. R*: Two seasons/years data generated at minimum two different agro-climatic zones
c. R**: Two seasons/years data generated at minimum three different agro-climatic zones
d. R#: One season / year data generated at minimum four different agro-climatic zones for fungicides,
insecticides and PGRs. Whereas, in case of herbicides, two seasons/years data generated at
minimum two agro-climatic zones in case of 9(3b) and two seasons/years data generated at
minimum three agro-climatic zones in case of 9 (3). In case of seed treatment one season / year data
generated at minimum three different agro-climatic zones will be required. In case of soil applied
Pesticides (Insecticide/ Fungicide/ Herbicide), if residue at harvest is above LOQ then 2 nd season data
will be required or else one season data will be sufficed.
Note: For commercial non edible crops ((like jute, jatropha, rubber, etc.,) only, the data on
residue and persistent is plant is not required.
e. R$: If Technical Import & Technical Indigenous Manufacture application which are submitted
together with Formulation Import & FormulationIndigenous Manufacture applications, no bio-efficacy
data required.
Note: In case of herbicides data on effect on soil Physico- chemical and biological properties and
effect on normally cultivated three succeeding crops is required along with residue studies in the
same plots of the field.
Example: for a herbicide intended to be registered for use in wheat crop data on effect on
succeeding crops of maize at location one, green gram at location two and sesamum at location three
may be generated along with residue studies. However, this is only an example and data on any other
normally cultivated succeeding crop may be generated.
1. For registration of Combination products of two / three registered pesticides (All sort of combination
should be addressed in this):data is required as per the guidelines of Formulation Import /
FormulationIndigenous Manufacture. However, data on Sr. No. 1, 2 (if any individualcomponent is
not registered as formulations, the two seasons comparable data are required to be generated against
target insects/ diseases/ weeds using stable / viable formulations. Already registered formulations of
individual component of the formulation at approved dosage shall also be included as check), 4,
(required only for insecticides and PGR)10, 11, 12, 13, 14, 15,16,17, 18 & 19 (Table – 1) if the a.i.
content of either component of the combination product is higher than the already registered same
formulations combinations, the applicant is required to submit the data.
Note: S. No. 14 & 15: Data on residue required for as many seasons and locations as required
in case of existing FIM u/s 9(3) guidelines. In case of herbicides data on effect on succeeding
crops and soil physico-chemical and biological properties are also required along with residue
studies as detailed in existing FIM under section 9(3) bioefficacy guidelines for herbicides.
S. No. 10,11,12,14 & 15: These data shall only be required when the concentration / a.i. dose
are higher than already registered and / or in case new compound is formed and / or any
adjuvants / diluents (other than water) / carrier is different that are in te registered
formulations of the individual components. Data on 12, 14, & 15 are also required for the
additional crop label claims to common of the individual registered.
2. If technical is already registered for import under section 9(3) and applicant want to apply for
indigenous manufacture of the same pesticide thenone season data on Sr. No. 1, & 2 (Table – 1) if
any, on two representative crops at two different agro-climatic zones is required and one season
residue data on two representative crops particularly on fruits and vegetables is required.
3. If formulation is already registered for import or for indigenous manufacture u/s 9(3) by any applicant
and the same registrant want to apply for technical indigenous manufacture, data on Sr. No. 1, 2 & 14
(Table – 1) not required. Whereas, if other applicants want to apply for technical indigenous
manufacture, one season data on Sr. No. 1 & 2 (Table – 1) on two representative crops at two different
agro-climatic zones is required and one season data on Sr. No. 14 (Table – 1) on two representative
crops particularly on fruits and vegetables is required.
4. In case of Technical Import from New Source,two seasons data on each crop mentioned in labels/
leaflets at least at two different agro-climatic Zones is required on Sr. No. 1 & 2 (Table – 1) and two
years or seasons data on Sr. No. 14 (Table – 1) on representative crops of each group on which
pesticide is approved.
Note: Data on Sr. No. 1 & 2 (Table – 1) is required on all registered formulations of same
technical on all approved crops at the time of Issue of import permit provided the application
for registration is received within 4 years of issue of import permit.
5. For registration of pesticide formulation for indigenous manufacture having the identical
chemical composition that of formulation already registered for import U/s 9(3).No data on
bioefficacy is required provided the technical of the source to be used in formulation is duly
registered as per guidelines of the Registration Committee and the label claims are same.
Whereas, for registration of pesticide formulation for indigenous manufacture having
different chemical composition that of formulation already registered for import u/s 9(3),
complete data on Bioefficacy to be submitted as per already existing guidelines for
formulation indigenous manufacture (FIM) U/s 9(3) provided the technical (source) to be
used for making formulation is duly registered as per guidelines of the Registration
Committee.
6. In case of Formulation Import without registering technical or Formulation Indigenous

Manufacturing without registering technical, in addition to data on formulation, complete
bioefficacy data on technical as per the guidelines of Technical Import or Technical
Indigenous Manufacture is required.
7. For Petroleum derived spray oil (PDSO), in case of 9(3) and 9(3b) data on Sr. No. 1, 2, 4, 13,
14, 15, 16 & 18 only as per Table – 1 is required.
8. For Manufacturing Use Product (MUP) of pesticide u/s 9(3) (Other than fruit ripening agent):
a. If the technical grade pesticide is not registered, complete data with respect to product
technical on Bio-efficacy as per guidelines for technical import u/s 9(3) to be submitted
along with data on MUP as listed below.
b. If the technical grade and source for import is duly registered as per guidelines of the
registration committee, no data on Bioefficacy will be required except Registration status
of MUP in foreign countries.
c. Rationale for import and registering the MUP to be submitted in Bioefficacy.
9. For use of Surfactant with registered pesticide Formulations, data ontwo seasons Sr. No. 1 & 2 (Table
– 1) from minimum three different agro-climatic zones are to be generated with surfactant (tank mix)
and without surfactant. The data on Sr. No. 10, 11, 12, 14 & 15 (Table – 1) are to be generated with
surfactant as per the requirement of the general guidelines u/s 9(3) of the Insecticides Act, 1968.
10. For registration of twin pack of two registered Herbicides and Surfactant, data onSr. No. 1, 2
& 13 (Table – 1) if proposed to mix, Sr. No. 10, 11 & 12, 14, 15, 18 & 19 (Table – 1) is required.
Note: Two season data on Sr. No. 1 & 2 (Table – 1) are to be generated on combination of the two
herbicides with & without surfactant and individual herbicide from minimum three different agro
climatic conditions. The bio-efficacy data on other parameters are to be generated on the combination
of the two herbicides with surfactant as per the requirement of general guidelines u/s 9(3) of the
Insecticide Act, 1968.
11. MRL fixation may be required for seed treatment products on those crops (such as
leafy vegetables, etc.) which are being consumed within one month of sowing.
Note:For Fungicides the efficacy trials should be conducted in areas where the claimed crop is the
main crop of that particular area and the area should be hot spot of the claimed disease i.e disease
pressure should be more than 30 % during the trial period and the fungicide should show minimum
70 % control. This must be certified by the SAU/ICAR institute where the study has been done. In
case of insecticide reduction over control should not be less than 70% and in case of herbicidesWeed
Control Efficiency (WCE) of individual weed species should be minimum 70%, which is duly
authenticated by ICAR/SAUs.
Data requirements for Registration for Post-Harvest Treatment (PGR) of crop produce:
a) Data requirements for Registration of Technical -
Note: The data should be submitted on Parameters for Bio-efficacy as per the guidelines
approved by the RC from time to time for registration of Chemical pesticides under TIM /
TI U/s 9(3) category, as the case may be. The specific data requirements / information for
registration of technical (TC/TK) are as under: -
(i) Fate and behavior in treated crop produce

(ii) Fate and behavior in air
(iii) Fate and behavior in water
(iv) Fate and behavior in soil & data on Sr. No. 16 & 18 (Table – 1) is required.
(v) If the product technical is not in gas form and/or also to be used to make formulation(s)
for use for other than fumigation of post-harvest crop produce (e.g. for use in field crops,
household purposes etc.), the additional data requirements on technical to be submitted
as per the guidelines approved by the RC from time to time for registration under TIM /
TI category U/s 9(3), as the case may be.
b) Data requirements for Registration of MUP (Manufacturing Use Product)(Not for Ripening agent)
–
Note: -
1) If technical grade of the pesticide is not registered, complete data with respect to product
technical on Bio-efficacy as per above guidelines under I. above for technical indigenous
manufacture (TIM) / technical Import (TI) U/S 9(3), as the case may be, to be submitted along
with data on MUP as listed below.
2) If the technical grade and source for import is duly registered as per guidelines of the
Registration Committee, data on MUP to be submitted as listed below:
(i) Registration status of MUP in other countries.
(ii) If the product MUP is not in gas form and/or also to be used to make formulation(s)
for use other than fumigation of post-harvest crop produce (e.g. for use in field crops,
household purposes etc.) the additional data requirements for registration of MUP to be
submitted as per the guidelines approved by the RC from time to time for registration of
Chemical Pesticides U/s 9(3).
c) Data requirements for Registration of formulation –
Note:
1) For registration of a formulation under FIM / FI U/s 9(3) category, the technical (TC/TK) and / or
MUP from which formulation to be manufactured, should be registered under the Insecticides Act,
1968.
2) An applicant seeking registration of a formulation without registering technical for

import/indigenous manufacture U/s 9(3), required to submit complete data as per the guidelines
for registration of formulation listed below, along with complete data as per above guidelines for
registration of technical for import/indigenous manufacture U/s 9(3) category, as the case may be.
3) The data parameters for Bio-efficacy shall be as per the guidelines approved by the RC from time
to time for registration of Chemical pesticides under FIM / FI U/s 9(3) category, as the case may
be. The specific data requirements / information for registration of formulation on post-harvest
crop produce (PGR) are as under: -
Table – 2
1. Data on bio-effectiveness For label claim of controlled atmospheric

Adverse effect on postharvest crop conditions data on 1. And 2. To be submitted
produce, if any (phytotoxicity, change in for threerepeated trials (same temperature and
appearance and flavour etc.) relative humidity)
OR
For label claim of ambient atmospheric
conditions data to be submitted for minimum
three repeated trials.
2. Residue and persistence at For label claim of controlled atmospheric
different interval (immediately after conditions (same temperature and relative
treatment, start from 0 hours till BDL or humidity) data to be submitted for three
acceptable level) in post-harvest crop repeated trials.
produce. OR
For label claim of ambient atmospheric
conditions (same temperature and relative
humidity) data to be submitted for minimum
three repeated trials.
3. Registration status of formulation in other Required
countries
4. MRL Performa and label leaflets Required
5. Label claims as per product Bio-efficacy information as per gazette notifications issued by Govt.
of India from time to time.
Note: Post harvest application of 1-MCP should be under expert supervision (PCO’s).
Remarks- Appropriate placement and comments from ICAR\NCIPM\SAU
1) Bio-efficacy data on target pests (diseases, insects, nematodes and weeds etc.) generated by
ICAR/SAUs and institutes under National Agriculture Research System and other institutes
approved by registration committee will only be acceptable.
12. Registration of Long Lasting Insecticide Impregnated mosquito bed nets and Long Lasting
Insecticide Incorporated mosquito net for registration U/S 9(3) :
1. Three years’ bio-efficacy trial in three locations. Out of three locations, two locations should be in
endemic areas. The bio-efficacy trial has to be conducted by adopting the protocol devised by the
Malaria Research Centre/VCRC (ICMR).
2. Baseline data on persistence of insecticides on the net and its analysis for comparison on yearly
basis.
3. Sustainability of fabrics.
13. Registration of insecticides for control of Ecto-parasites (Mites, Bedbugs, Ticks etc.) in poultry u/s 9(3)
& 9(3b):
a) Technical 9(3) & 9(3b) – Data on parameters on Table 1 at Sr. No. 7, 8, 10, 11, 16 & 18 are
required.
b) Formulation 9(3) & 9(3b) – Data on parameters on Table 1 at Sr. no. 13, 16 & 18 are required. In
addition to Table 1 at Sr. no. 13, 16 & 18 data onEffect on layers (duration 3 months, study to
commence preferably at the age of 40 weeks) – data generated in National/ ICAR/SAU
Laboratories [three for 9(3) and 2 for 9(3B) ] on [ (i) change in body weight, (ii) feed intake, water
intake, Feed Conversion Ratio (FCR) (iii) mortality and morbidity pattern, (iv) * Clinical
symptoms/morphological changes in organs, (v) * Blood profile and Enzymology,(vi) egg
production records for 15 days.], Persistence and residue on treated surface, Residues in various
organs of birds and edible products, Residue in birds excreta. (Rearrange)
Note: In case of import of formulation without registering technical, whole set of data on technical
shall be submitted along with the application.
14. Registration of insecticidal formulations for use in aircraft disinfection:
1. All insecticides for aircraft disinfection must be manufactured only from the technical grade
insecticides which are registered under the Insecticides Act, 1968
2. Data on insecticides formulation shall be considered along with the data on technical grade
insecticides and not in isolation.
Bioefficacy and residue data:Bioefficacy test on the proposed formulation should be conducted in
Indian conditions minimum 2 trials in each of 3 National Laboratories / NABL accredited
laboratories, recognized by Government of India.
Data on persistence of pesticides on commonly used surfaces in the aircrafts and concentration in
air, as applicable, should be generated in three National Laboratories/ NABL accredited laboratories.
15. Data requirement for registration of insecticides for use in public health programme u/s 9(3):
1. Bioefficacy data generated by ICMR / MOH&FW Institutes based on multi-centric three

years / seasons as per their protocol.
16. Data requirements for registration of house hold pesticides :
a. All household pesticides as defined must be manufactured only from the technical grade pesticides
which are registered under the Insecticides Act, 1968.
b. Data on household pesticides formulation shall be considered along with the data on technical
grade pesticides and not in isolation.
A. Bio-efficacy claims In case of Formulation:
a. Bio-efficacy claims to be given on the labels as under:
(i) A brief direction concerning the major usages of the pesticides should be given on the
labels.
(ii) Whenever the Registration Committee has approved the product for restricted use, this
should be indicated very clearly on the labels in capital letters. ‘For use only.
(iii) Instruction regarding Insecticide ‘Not to be used on any food crop to be given’.
b. Bio-efficacy claims to be given on the leaflets:

(i) Detailed information on the usages of insecticide indicating the name of insects, method
of application, dosage, places of treatment, PP equipments to be used etc. should be given
in paragraphs forms. Common name of the insects should be given.
(ii) Whenever the registration committee has approved the product for restricted use, this
fact should be indicated very clearly on the leaflets in bold letters.
(iii) Instructions regarding Insecticides ‘Not to be used on any food crop to be given’.
B. Bio-efficacy claims to be given on the labels and leaflets in case of technical grade material:The
purpose of import /manufacture of technical grade material is required to be given on the labels and
leaflets.
C. Data requirements on Bioefficacy and Residues for formulation of pesticides for provisional
registration of U/s 9(3B).
(I) The applicant should submit published / cited Indian data on bio-effectiveness in support
of the claims indicated on the labels / leaflets. The data should be produced from 2
National laboratories based on minimum 2 repeated trials. This should be further
supported with any published information available from elsewhere (overseas data).
(II) Information on secondary pests outbreaks particularly of ticks and mites should be
given where residual pyrethroids are being used.
(III) Data on Residues:Data on persistence, of the pesticides which should be on different
types of surfaces should be submitted / generated obtained under foreign / Indian
conditions from 2 laboratories. This may also be supported by data generated elsewhere.
(IV) Data on concentration of a.i. in Air – for Aerosols (e.g. Coil, mats, liquid vaporizer
etc.). Registration Status in foreign countries.
D. Data requirements on Bio-efficacy & Residues data requirement for regular registration
of formulations of pesticides U/s 9(3):
i. Bio-efficacy test on the proposed formulations should be conducted under Indian

conditions minimum on two trials in each two national laboratories with three
replications. In case major difference is reported then data from third National
laboratory is required.
ii. Data on persistence of pesticides on different types of surfaces should be generated in
three national laboratories wherever applicable.
iii. Information on secondary pests outbreaks particularly of ticks and mites should be given
where residual Pyrethroids are being used.
iv. Data on concentration of a.i. in Air – for Aerosols (e.g. Coil, mats, liquid vaporizer etc.).
(Added based on approval in 266th meeting of RC held on 20-07-2006)
v. Registration Status in foreign countries.
E. Data requirements for the registration of new formulations of the approved

pesticides:Data on bio-effectiveness and persistence on different surfaces as required in case of
Registration of formulations for regular registration under section 9(3).
F. Data required for combination products:Bio-efficacy data on combination products v/s
individual products. Data on persistence on different types of surfaces should also be submitted.
All above data should be generated as per the requirement indicated in case of pesticides
required for regular registration under section 9(3).
G. Methodology:
Flying/crawling insects:
Residual films of insecticides prepared by spraying insecticides on different types of surfaces,
such as Glass, Wood, Mud & Cement surfaces. Insects to be exposed for 30 minutes and then
shifted to recovery chambers for 24 hours after which the mortality count should be made and
the satisfactory mortality of insects would be more than 90%. The residual toxicity of
insecticides should also be studied at different intervals. Evaluation of space spray against
flying insects should be conducted in PEET GRADY Chamber as per standard ISI
specification 1824, mats/coils could also be evaluated inside the Peet Grady Chambers against
caged mosquitoes and the knock down effect is to be recorded at different intervals. Aerosols
are to be evaluated inside a standard room. The test is to be conducted as per WHO technical
reports series No. 206.
C. TOXICITY
Sl. Parameters Technical Formulation Technical/

No. Formulation
9(3b) 9(3) 9(3b) 9(3) 9(4) 9(4)
TI/TIM TI/TIM FIM FIM/FI TIM TI/FIM/FI

1. Acute oral Rat R R R R NR NR
2. Acute Dermal- Rat/ R R R R NR NR

Rabbit
3. Acute inhalation-Rat R R R R NR NR
4. Primary Skin R R R R NR NR
Irritation-Rabbit
5. Acute Eye Irritation- R R R R NR NR
Rabbit
6. Skin Sensitization R R R R NR NR
Test–Guinea Pig
7. Repeated dose range R R NR/R NR/R NR NR
finding oral toxicity
study (28 days)*-Rat
8. Repeated dose 90 R R NR/R NR/R NR NR
days oral (Rat)
9. Repeated dose 90 days R R NR NR NR NR

oral toxicity-Dog***
10. Repeated dose dermal R R NR/R NR/R NR NR

toxicity-Rat/Rabbit
11. Repeated dose R R NR/R NR/R NR NR
inhalation toxicity*#-
Rat
12. Acute Neuro-toxicity- R R NR NR/R NR NR
Rodent*
13. Repeated dose R R NR NR/R NR NR

Neurotoxicity-
Rodent*
14. Delayed R R NR NR/R NR NR
Neurotoxicity- OP
compound- Acute
exposure-Laying Hen
15. Delayed R R NR NR/R NR NR
neurotoxicity- OP
compound- Repeated
Administration-
Laying Hen
16. Developmental R R NR NR/R NR NR

Neurotoxicity*-
Rodent
17. Combined R R NR NR NR NR
carcinogenicity &
chronic toxicity-Rat
18. Carcinogenicity-Mice R R NR NR R NR
19. Developmental NR R NR NR NR NR
toxicity study-
a) Rat &
b) Rabbit
20. Two generation NR R NR NR NR NR

reproduction toxicity
in Rat
21. Mutagenicity ** NR R NR NR NR NR
22. Pharmacokinetics and R R R R NR NR

Metabolism in Rat. (Two (Two (One (One
species) species) species) species)
23. Feeding study R R NR NR NR NR
including Metabolism
in Livestock (Goat,
cow)/Poultry, Hen
24. Pharmacokinetics and R R NR NR NR NR
Metabolism in other
mammals and its
similarities or
differences from
humans.
25. Immunotoxicity R R R R NR NR
study*
26. Acute Avian toxicity * R R R R NR NR

27. Repeated dose Avian R R R R NR NR
toxicity (One species)
28. Avian Reproduction R R R R NR NR

toxicity (One species)
29. Acute toxicity to Fresh R R R R NR NR

Water Fish (One
species)
30. Acute Toxicity- R R R R NR NR
Honey Bee (Oral &
Contact)
31. Acute Toxicity- R R R R NR NR

Earthworm
32. Medical data R R R R NR NR
33. Human toxicity NR NR NR R NR NR

information from
foreign countries
34. Health records of NR R NR R NR NR

Industrial workers
35. International report on NR R/NR NR NR NR NR

Carcinogenicity &
Genotoxicity
Footnotes:
Toxicity study No.9- Repeated dose 90 days oral toxicity study (Dog): ***Peer reviewed
international published literature is also acceptable for dog study.
Toxicity study no.17,18
i. The Combined Carcinogenicity and Chronic Toxicity study in Rat and

ii. Separately carcinogenicity study in mice is required.
23)- No separate footnote is required because feeding and metabolism study can be done together
Some metabolites can be more harmful than parent compound even in very small quantity due to bio-
activation so it can be conditional requirement, depending on case to case basis.
1) Technical (TI and TIM) u/s 9(3b) and 9(3) :-In case of Technical, name and Percentage of relevant
and toxic impurities and/or metabolites should be indicated. Also data/information should be
provided about their toxicity.)
2) Technical u/s 9(3):-
a. TI u/s 9(3):- In case of Technical Import, data from Sl. No. 1 to 35 are required.
b. TIM u/s 9(3):- In case of Technical Indigenous Manufacture, data of Sl. No. 33 and S. No. 34
are not required.
3) Technical Import from new Source u/s 9(3):- Data on parameters 1 to 11;21, 32-33 and 35 is
required.
4) Technical Indigenous Manufacture u/s 9(3) in case same technical is registered for import or
formulation made from the same technical is registered for import or indigenous manufacture:
I. If impurities are identified, quantified and impurities are within limits (i.e. within
maximum of already registered technical),then data on parameters 1-6 and 21 is required.
II. In case impurities which are not toxic and not relevant and are not within maximum
limits of registered technical however are within +3%) and no new impurity is there,
then same data as above (Parameters 1 to 6 and 21) will be required.
III. If impurities are identified, quantified and are not within limits of registered technical or
if impurities are not within +3%) and/or any additional impurity is present then in
addition to the tests as indicated in (4) above, additional tests based on the nature and
quantity of impurity and QSAR alert will be taken on case to case basis.
5) Technical indigenous manufacture u/s 9(3) in case same technical is registered for import or
formulation made from the same technical is registered for import BY THE SAME
APPLICANT WITH SAME COMPOSITION, PROCESS OF MANUFACTURE ETC.:- Data
on Ames Test only is required.
6) 9(4) TIM:-In case of Technical indigenous Manufacture u/s 9(4), data on Sl. No. 21 data of @AMES
Test (Tier-I) is required.(Only AMES test is required)
7) Formulation u/s 9(3b):Data as per FIM u/s 9(3b) required as indicated in table above.
8) Formulation u/s 9(3):-For FIM :- as indicated in table except data of Sl. No. 33 and S. No. 34 are
not required.
9) In case of FI WRT (Formulation Import without registering Technical) OR FIM WRT
(Formulation Indigenous Manufacture without registering Technical):-in addition to data on
formulation, complete toxicity data on technical as per the guidelines of TI , orTIM ( as the case may
be) should also be submitted.
10) 9(4) TI/FIM:-No data is required, in case of Technical Import(TI) or Formulation Indigenous
Manufacture (FIM) u/s 9(4) application.
11) For LLIN:Acute toxicity studies (Six pack) i.e. S.No. 1 to 6 are required with Premix and health
monitoring studies in users as per approved protocol is required with final product (LLIN).
12) MUP:
a) Data on the parameters from S.No. 1 to 6, should be submitted.
b) If the Technical grade pesticide from which MUP is to be prepared is not registered, complete
data with respect to product Chemistry (along with sample and reference standard of technical
grade pesticide and impurities), Bioefficacy, Toxicity and Packaging as per the applicable
guidelines for registration of technical should be submitted.
13) Household Pesticide Formulations:
a) Data on the parameters from S.No. 1 to 6, should be submitted for pesticides in Solid &
Liquid form. In case of Pesticides in Vapour form or which emits vapour/fumes, in addition to
parameters from S.No. 1 to 6, Health monitoring study of the user by using the household
pesticides in its actual use should also be submitted. The study should be as per the protocol
approved by the RC.
b) Data on household pesticides formulation shall be considered along with the data on technical
grade pesticides and not in isolation.
14) Pesticide Formulation for use in Public Health :Data on the parameters from S. No. 1 to 6, 26, 27
& 29 should be submitted. Recommendations from National Vector Disease Control Program, M/o
Health & Farmers Welfare are also required.
15) Formulation for use for Aircraft Disinfection:If the technical grade is duly registered as per
guidelines of the Registration Committee, data on the parameters from S.No. 1 to 6 should be
submitted.
16) Herbicides in twin pack with no other herbicide or with surfactant:MSDS and Acute toxicity
information on surfactant should be submitted along with data on herbicide formulation.
* REFER TO GUIDANCE DOCUMENT ON TOXICOLOGY FOR REGISTRATION OF

CHEMICAL PESTICIDES IN INDIA.
** In Mutagenicity test; an Ames test, any two in-vitro and one in-vivo Mutagenicity tests are required.
#
Repeated Dose Inhalation toxicity Study for 90 days exposure would be required if there is likelihood
of significant repeated inhalation exposure as in case of gas, vapours , aerosols, fumigants or
likely duration of human exposure via inhalation is long viz. Mosquito coils; sprays used
repeatedly.
Note: The general recommendations as mentioned in the Guidance Document on Toxicology for
registration of Chemical Pesticides in India will be applicable on the basis of merit case to case
basis.
Waiver would be considered only when existing information provides robust and full scientifically
sound weight of evidence approach and read across/bridging from structurally and /or biologically
related similar pesticides specifically case to case basis on full merits.
The replacement alternatives not involving experiments on animals would not be normally
considered,except in exceptional and rare cases where in case of alternatives if available with full
and sound justification is provided specifically case to case basis on merit subject to full
satisfaction of the expert regarding full toxicity data provided for the alternatives.
D. PACKAGING
Sl. Parameter Section 9(3B) Section 9(3) Section 9(4)

No.
TIM FIM TI TIM FIM TI FI TIM FIM TI FI
1. Labels and Leaflets per R R R R R R R NR NR NR NR

IR-1971, all fields (as
applicable) and as
amended from time to
time
2. Manner of labeling and R R R R R R R R NR NR NR
Leaflet
3. Type of packaging R R R R R R R R NR NR NR
(Ultra small, small or
Big whichever is
applicable)
4. Manner of packaging R R R R R R R R NR NR NR
5. Specification for R R R R R R R R NR NR NR
primary, Secondary and
Transport packages
(whichever is
applicable)
6. Details of packaging R R R R R R R NR NR NR NR
material and its
compatibility with
content
7. Performance of R* R* R* R R R R NR NR NR NR
container with content
during storage stability
test(Shelf life Study)
8. Transport worthiness R* R* R* R R R R NR NR NR NR
test
Chapter V of the Insecticides Rules 1971 in the Insecticides Act, 1968, the rule 16 to 20 of the said
chapter deals with the Packaging and Labeling.
R*- Before Commercialization the data will be required.
Note:
1. In case of additional packaging endorsement applications (already approved packaging), the data at Sl.
No. 05, 06, 07& 08, are not required if similar packaging (material) and manner of packaging is being
sought by the applicant as has been granted to earlier 9(3) registrant.
2. Specification of Bureau of Indian Standard (BIS) must be followed for all the packaging requirements
(Wherever available and applicable).
3. All Packaging tests must be carried out with the product of same batch and in its commercial package
preferably in Indian condition.
4. The duration of the test and the conditions including geographical conditions must be mentioned.
5. Storage stability data should be generated keeping at least the following parameters in test protocol
such as test temperature, test duration, test packaging material, content of active ingredient (a.i.) in the
product during and after storage, test humidity, exposure to light, physical and chemical properties of
the product during and after storage etc.
6. The testing protocols must have their basis in the WHO/FAO/ CIPAC/ASTM recommendations or
other validated methodology of GLP/ NABL accredited laboratory having packaging testing (chemical
/ mechanical as applicable etc.) in the scope.
7. The Accelerated Storage Study (ASS) test must be conducted at 540C ± 2oC (wherever applicable)
containing corrosiveness study of the product in reference to packaging material for 14 days as per
FAO/ WHO manual for claiming appropriate shelf life of the product which can be maximum two
years, subject to the condition of providing the ambient storage stability study data of thirty months or
as the case may be within thirty months from the date of application for the registration.
ANNEXURE- III
DATA REQUIREMENTS FOR MICROBIAL PEST CONTROL AGENTS (MPCA)

PESTICIDES
Bio-pesticides/ Microbial Pest Control Agents (MPCA)
1. The applicant needs to submit MOU/license agreement between the applicant and the
inventor (either own R&D Laboratory or outsourced Research Institute/Facility) or
Authorization letter from the inventor of strain OR undertaking by the applicant about the
name of inventor/source of strain as perAnnexure-I.

2. Undertaking declaring that the product is free from Chemical pesticides/Botanicals
pesticides/Other Agro-Chemicals asAnnexure-II.
3. Undertaking on bio-pesticides composition asAnnexure-III.
4. Updated Stakeholder list for all members in Association/Organization claiming for
MOU/authorization for data/technology utilization for mass
multiplication/commercialization of the strain, ifapplicable.
5. Form-I dully filled and signed giving complete details along with requisite fee as
applicable.
6. Notarized copy of BOD Resolution/ affidavit in case of proprietor/ partnership deed in case
of partnershipfirms.
7. Correct composition as per earlier 9(3) / 9(3B) registrant of bio-pesticidestrain.
8. The applicant should also, submit notarized copy of the Permanent Account No. (PAN),
allotted by the Income TaxDepartment.
9. In case of company, the Certificate of Incorporation granted by the Registrar ofCompanies.
10. Fee receipt issued by NBAIM, Maunath Bhanjan, UP after submission of sample for DNA
verification.
11. Undertaking that the product is free from GMO asAnnexure-IV
The Data Requirements for Microbial Pest Control Agents (MPCA) Pesticides
i) Entomopathogenic/ EntomotoxicBacteria
ii) AntagonisticBacteria
iii) EntomopathogenicFungi
iv) AntagonisticFungi
v) Nuclear Polyhedrosis Virus (NPV) & Granulosis Virus (GV)
A. Chemistry:
Sl. Characteristics Microbial (Antagonistic bacteria,

No. Entomopathogenic/Entomotoxic
bacteria, Entomopathogenic fungi,
Antagonistic fungi, and Baculovirus
Primary/Mother Formulated product

Culture
9(3B) 9(3) 9(3B) 9(3)
1. Systematic name (Genus and species) R R R R
2. Strain name R R R R
3. Common name, if any R R R R

4. Source of origin R R R R
5. Specification of the product containing R R R R
Habitat, Physical appearance and
morphological description, pH, particle
size, suspensibility, miscibility etc
parameters
6. Isolation of strain &Manufacturing R R R R

process
7. Methods of analysis including R* R* R* R*

Quantitativeanalysis
8. Shelf life claims R R R R
9. Data on storage stability as per shelf NR R NR R

lifeclaims
10. Composition of the product R R R R
11. Potency of product by bioassay method NR# NR# NR# NR#

(LC 50 (Beta, Delta , Cry toxin
endotoxin content, classification(delta
endotoxin)
12. CFU/g or ml R R R R
13. POB/Capsule count per ml/g of the R% R% R% R%

product
14. Adjuvants NR NR R R
15. Human pathogens (culture method) R R R R
16. Percent content of the Bio-control R R R R

mass/organism in the formulation &
nature of biomass.
17. Percentage of carrier/filler, wetting/ R R R R

dispending agent, stabilizers/
emulsifiers, contaminants/ impurities
etc.
18. Moisture content R R R R

19. Contaminants:
R R R R
Pathogenic contaminants such as
Salmonella, Shigella, Vibrio and such
other microbial, not to exceed 1 x 104
count per ml or per g of formulation.
20. Undertaking for free from Chemical R R R R

and botanical pesticide contaminants
21. Natural occurrence of the organism R R R R
22. PCR / Immunology assays ELISA Test NR$ NR$ NR$ NR$
23. Separation and purification of crystals NR NR NR NR
24. A sample for verification (500 @ g

or
500@ mL as the case may be)
a. DNA fingerprinting for the strain R R R R
verification from NBAIM, Mau
Bhanjan.
b.Pre-registration verification at Central R R R R

Insecticide laboratory (CIL)
 *Test procedure and criteria used for identification – morphology, biochemistry, serology/
Immunology for Entomotoxicbacteria.
 # and $ these parameters are required for Entomotoxic bacteria.

 % these parameters are required for Virus.
 @ Two samples of same batch of 500 gm/ml each along with copy of the Fee receipt shall be
submitted to Central Insecticides Laboratory, Faridabad for PRV purpose by the applicant for
Entomotoxic bacteria.
 POB/Capsule count per ml/g of the product only forNPV.
 Viral unit: NPVs 1x109 POB/ml or gm. minimum, GVs: 5x10 9 capsules /ml or gm
minimum.
 Dual culture to attain at least 50% reduction in target organism (35% for antagonistic
bacteria). Bioassay: based on diseased severity and rootcolonization.
 Natural occurrence of the organism, Immunology assays: Elisa and Separation and
purification of crystals are required for Entomotoxicbacteria.
 Test procedure and criteria used for identification by DNA test (Restriction enzymes
analysis test).
 Biological assays for determining the LC50 / LD50 of the formulation for
Entomopathogenic Viruses. Production of Entomopathogenic Viruses at commercial-
scale was done exclusively in-vivo by culturing large number of larvae of host insect
and subsequently feeding them with semi-synthetic diet contaminated with virus
inoculums in laboratory. Viruses’ production in-vitro by culturing insect cells in
bioreactors was a substitute for labor intensive maintenance of the massive host-insect
colony.
 Manufacturing process including type of fermentation and biological end products.
The microbial cultures are multiplied by liquid solid fermentation. Information
pertaining to use of entire mycelia mats with spores separated must be provided in
terms of biomass.
Documents to be mandatorily furnished by applicant applying u/s 9 (3)/ 9(3b) for all
categories of bio pesticides
1. Verification of the Authorization letter submitted by the applicant via e mail by

Secretariat from the original inventor/source of the strain for data utilization for mass
multiplication.
2. MOU/license agreement between the applicant and the inventor (either own R&D
Laboratory or outsourced Research Institute/Facility) or Authorization letter from the
inventor of strain OR undertaking by the applicant about the name of inventor/source
of strain as per Annexure-I
3. Updated Stakeholder list for all members in Association/ Organization claiming for
MOU/authorization for data /technology utilization for mass
multiplication/commercialization of the strain.
4. Relevant Affidavit/Undertakings:-
a) Affidavit on bio-pesticide composition on NJSP duly notarized.
b) Notarized copy of depositing microbial bio-pesticides strain sample in
National Repository with reference code number.
c) Undertaking on NJSP duly notarized that product do not contain any
genetically modified organism in the prescribed format.
d) Undertaking on NJSP duly notarized that product is free from

chemical/botanical pesticides / other agro-chemicals.
e) Affidavit of strain Innovator or applicant
f) Copy of 9(3B) Registration certificate, if relevant
Note:
1. Bt products should be labeled with bio potency and (or) toxin content. In addition, the
labels will have to contain a measurement of toxin protein as percent protein, referring to
the Lepidopteran-active toxin(s) present in thecrystal.
2. The presently used Bt var. kurstaki standard is HD-1-S-1980 and its potency was calculated
at 16,000 IUs per milligram of powder (Beegle et al. 1986. Standardization of HD-1-S-
1980: US Standard for Lepidopterous-active Bacillus thuringiensis. Bulletin Ent. Soc.
America 32: 44-45.). This standard strain is now available with PDBC, Bangalore and
DOR,Hyderabad.
3. Defined potency and toxin concentration – Bioassay would require the use of an insect
species. Normally manufacturers could select Trichoplusia ni / Helicoverpa armigera for
Lepidopteran specific Bt formulations. Spodoptera Units (SPU), Leptinotarsa Units
(LTUs) or International Toxin Units (ITUs) are to be used for denoting a specificinsect.
4. No test for beta exotoxin is required for Bacillus sphaericus, because this species is not
known to produceexotoxins.
5. The biopotency of products based on B. thuringiensis subsp. israelensis (Bti) is compared
against a reference strain IPS82, 1884 using early fourth-instar larvae of Aedes aegypti
(strain Bora Bora). The toxicity of IPS82 has an arbitrarily assigned toxicity of 15,000
ITU/mgpowder.
6. The biopotency of products based on B. sphaericus (Bsh) is determined against a reference
standard SPH88, strain 2362 using early fourth-instar larvae of Culex pippins (strain
Montpellier). The toxicity of SPH88 has an arbitrarily assigned toxicity of 1,700 ITU/mg
of the powder (Guidelines for laboratory and field testing of mosquito larvicides, WHO
2005 pp45).
7. The use of alternative bacterial reference powders and / or strains must be approached
cautiously. Such alternatives must be the subject of careful cross- calibration against the
reference powders and should be conducted by recognized laboratories and should be made
available to anyone who wishes to use, or check, the test with the alternative
powders/strains.
8. Water content should not exceed 8 %, (12% in Pseudomonas spp) to preclude premature
degradation of the product.
Guideline for already registered formulation/Strain u/s 9(3) under the IA, 1968.
Applicant shall submit only one folder containing the following documents:
I. Form-I dully filled and signed giving complete details along with requisite fee as
applicable.
II. Notarized copy of BOD Resolution/ affidavit in case of proprietor/ partnership deed in
case of partnership firms.
III. Correct composition as per earlier 9(3) / 9(3B) registrant of bio-pesticide strain.
IV. The applicant should also, submit notarized copy of the Permanent Account No. (PAN),
allotted by the Income Tax Department.
V. In case of company, the Certificate of Incorporation granted by the Registrar of
Companies.
VI. Authorization letter from the inventor of strain OR undertaking by the applicant about
the name of inventor/source of strain as per Annexure-I.
VII. Requisite number of stamp and envelopes.
VIII. Copies of Label Leaflets of the product as approved by RC of already registered strain.
IX. Copy of letter of Accession No. of strain or information on Accession number of strain.
X. Undertaking declaring that the product is free from Chemical pesticides/Botanicals
pesticides/Other Agro-Chemicals as Annexure-II
XI. One sample of 500 gm/ml quantity shall be deposited to NBAIM, Maunath Bhanjan for
test relating to DNA fingerprinting particularly partial gene code sequencing of desired
strain and a fee may be paid directly to NBAIM through DD oronline.
XII. Original fee receipt issued by NBAIM, Maunath Bhanjan, UP.
XIII. Undertaking that the product is free from GMO asAnnexure-IV
XIV. Undertaking on bio-pesticides composition as Annexure-III
XV. Toxicology data shall be accepted from Non-GLP laboratory also for encouraging of
new strain registration/any new or repeat studies for old strain. This decision shall be
applicable to all categories of biopesticide registration, henceforth. The applications
under scrutiny in the Secretariat of CIB&RC are also covered under this decision.
XVI. A sample of 500 gm/ml shall be deposited in the Secretariat of CIB&RC along with
File/documents for PRV purposes.
XVII. The above folder shall be scrutinized by the Chemistry division of the Secretariat of
CIB&RC.
XVIII. No preliminary scrutiny is required for applications for already registered strain of
bio- pesticides.
XIX. A letter may also be written by the Secretariat of CIB&RC to the Director, NBAIM,
Mau Nath Bhanjan, UP for submitting the DNA finger print report directly to the
Secretariat of CIB&RC, certifying that the DNA of strain submitted by the applicant
(Strain No.) matches with original Strain or otherwise.
XX. RC also decided that any government laboratory willing to undertake such studies on
the terms and conditions as approved by the committee may request Secretariat of
CIB&RC so as to seek approval from RC.
Annexure-I
AFFIDAVIT ON BIO-PESTICIDE STRAIN BY INVENTOR OR APPLICANT
I, …………………….. S/o ……………………………………, aged …………….years, resident

of
………………………………………………..and Proprietor/Authorized person of the firm
M/s
………………………………………………………………….., having its office
at
……………………………………………………….. do hereby declare and solemnly affirms as
under:
That I am in the capacity of …………………………….. of firm M/s

……………………………… do hereby declare that the information furnished with respect to
composition in Form-I, Label/Leaflet and bonafide verification of the application fir registration
of (Name of the product)…………….., CFU/PBO…………….per gm or ml min; Strain No.
(Name and number of registered strain)…………. invented by M/s. …………………………is
registered under section 9(3b) or 9(3) of the Insecticides Act, 1968.
1. That a shelf life of the product shall be twelve/Six/four months.

2. The product shall be packed as per IS: 8190 (Part-I) 1988 for Solid Pesticide (Second
Revision).
3. That there will be no change in chemical composition, shelf-life, packaging requirement
and the product will have the quality and packaging as per the relevant IS or as per
specification approved by Registration Committee for 9(3b) & 9(3)registrant.
I, shall be responsible for adhering to the above composition and strain while manufacturing and
marketing the product for distribution or sale. In case of any violation of the above declaration
and also the conditions laid down on the Certificate of Registration of the said Bio-Pesticide,
interalia, Product Quality Speciation submitted by us and also to the specification as and when
the same are formulated and published by BIS amendments thereof, I am liable to be
prosecuted/rejection of application under the provisions of Insecticide Act, 1968 and the Rules
1971 and amendments thereof.
Deponent
VERIFICATION
I, ……………………………., the above deponent do hereby verify that what has been declared
above is true to the best of my knowledge and belief and nothing has been concealed there from.
Deponent
Annexure-II
UNDERTAKING FOR ABSENCES OF CHEMICAL/ BOTANICAL PESTICIDES/
CONTAMINATS/OTHER AGRO-CHEMICALS;

of ………………………………………………..and Proprietor/Authorised person of the firm
M/s ………………………………………………………………….., having its office at
……………………………………………………….. do hereby declare and solemnly affirms
asunder:
That the product, (Name of the product)……………………….. formulation containing

CFU/PBO/Delta endotoxin content.…………. per gm or ml min, Strain code, (if
any)…………..Strain No: ……………. manufactured by (Name of the applicant)………………
does not contain any Chemical/Botanical Pesticide/Contaminants/other Agro-Chemicals.
That I/we shall provide the samples of our product (Name of the product)
……………………………. as and when desired by the competent Authorities of Government of
India for verification.
That my/our above undertaking is true, and no portion is false and I have concealed nothing
relevant to the above matter.
Place & date: Deponent Signature:

Name Designation: Company Seal:
Annexure-III
AFFIDAVIT ON BIO-PESTICIDE COMPOSITION
of ………………………………………………..and Proprietor/Authorised person of the firm M/s
………………………………………………………………….., having its office at
……………………………………………………….. do hereby declare and solemnly affirms
asunder:
That I am in the capacity of …………………………….. of firm M/s
……………………………… do hereby declare that the information furnished with respect to
composition in Form-I, Label/Leaflet and bonafide verification of the application for registration
of (Name of the product) …………….., CFU/PBO…………….per gm or ml min; Strain No.
(Name and number of registered strain)…………. under section 9(3b) or 9(3) of the Insecticides
Act, 1968 is as under:-
1. COMPOSITION: (SPECIMEN FOR Pseudomonas fluorescence WP) Components:
A. Quantity : (%w/w)
a) Pseudomonas fluorescence : CFU 1x108 CFU/gm min1.0%
b) Carboxy methyl cellulose : 1.0%
c) TalcPowder : 98.0%
Total : 100.0%
2. That a shelf life of the product shall be twelve/Six/four months.
3. The product shall be packed as per IS:8190 (Part-I) 1988 for Solid Pesticide (Second
Revision).
4. That there will be no change in chemical composition, shelf-life, packaging requirement
and the product will have the quality and packaging as per the relevant IS or as per specification
approved by Registration Committee for 9(3b)registrant.
5. Bonafide declare that M/s ………………………………., manufacturing premises
proposed/located at ……………………………………………… having Registration Certificate
total no., if any or Nil and manufacturing license no. if any or Nil.
I shall be responsible for adhering to the above composition while manufacturing and marketing
the product for distribution or sale. In case of any violation of the above declaration and also the
conditions laid down on the Certificate of Registration of the said Bio-Pesticide, interalia,
Product
Quality Speciation submitted by us and also to the specification as and when the same are
formulated and published by BIS amendments thereof, I am liable to be prosecuted/rejection of
application under the provisions of Insecticide Act, 1968 and the Rules 1971 and amendments
thereof.
Deponent
VERIFICATION
I, ……………………………., the above deponent do hereby verify that what has been declared
above is true to the best of my knowledge and belief and nothing has been concealed therefrom.
Deponent
Annexure-IV
UNDERTAKING BY MANUFACTURERS OF MICROBIAL PESTICIDES
I,------------,aged-------years, s/o-----------, R/o-------------an------------of M/s.------------Registered

Office at-----------do hereby undertake as follows:
That the product--------------based on------Strain No.-------, manufactured by M/s.------and /or
imported by M/s…………..does not contains any genetically modified organism (GMO) .
a) That I/We shall abide by the provisions contained in the International Plant
Protection Convention with regard to the import of this product.
b) That I/We shall abide by the provisions in context of International Standards for
Phyto-Sanitary Measures-Code of Conduct for the import and release of exotic bio-
pesticidesoftheInternationalPlantProtectionConvention(IPPC),FAO,Rome/Plant
Quarantine (regulation of Import into India) order,2003.
c) That I/We shall provide the samples of our product as and when desired by the
competent authorities of Government of India for verification.
d) That I/We further undertake that in the event of the above product having proved
otherwise by any competent authority and resulting in environmental damage, I/We
shall inform to Plant Protection Adviser, Dte. of PPQ&S, Sectt. of Central
Insecticides Board and Registration Committee, and other relevant authorities for
Manufacturing Licensing, Pollution Control and of appropriate
District/State/National Level and shall comply with the directions from them.
e) That my/our above undertaking is true, and no portion is false and I have concealed
nothing relevant to the above matter.
Signature:----------------
Date------- Name-------------------
Place:---------------- Designation-&Seal of the Company-----
Bio-efficacy
Sr. Particulars Primary Formulated

No. culture/mother product
culture
9(3B) 9(3) 9(3B) 9(3)
1 Field studies: NR NR R** R***
Data on bio-effectiveness and phytotoxicity
generated at ICAR, SAUs, CSIR / ICMR
institutes. The data should be certified
either by the Director or Head of the
Institute.
2 Laboratory studies: R R R R
The product should be tested at a
laboratory under ICAR/ SAU/
CSIR/ICMR.
2.1) LC50 values for each insect species

under laboratory conditions should be
generated at least at two institutes of ICAR,
SAUs, CSIR and ICMR.
2.2) Data on LC50 values for each target

insect species should be generated at a
laboratory under ICAR/ SAU/CSIR/ICMR
3 Data on non-target organism: One NR NR R R

season/one year on effect on product against
natural parasites/ predators.
 R = Required, NR = Not Required

 R** - Two seasons/years data on bio-effectiveness from two agro-climatic Zones
 R *** - Two seasons/years data on bio-effectiveness from minimum three agro climatic Zones.
 2.1) - Applicable for Entomotoxic Bacteria

 – Applicable for NPV & GV.
 Sr. No. 3 - Required in case of Entomopathogenic fungi, Entomopathogenic Bacteria.
Note: No bio-efficacy data required for already registered strains of Bio-pesticides.

Certificate of Registration will be granted as per approved formulation u/s 9(3)
Toxicity
S. No. Parameters Microbial (Antagonistic bacteria,

Entomopathogenic/Entomotoxic bacteria,
Entomopathogenic fungi, Antagonistic fungi,
and Baculovirus
Primary culture/mother Formulated product

culture
9(3b) 9(3) 9(3b) 9(3)
1. SingleDose Oral – Rat R R R R
(Toxicity/Infectivity/Pathogenicity)
2. Single Dose Dermal – Rabbit R R R R
3. Acute Inhalation (a) R R R R
4. Single Dose Pulmonary –Rat (b) R R R R
5. Single Dose Intraperitoneal–Rat (c) R R R R
6. Single dose intravenous (d) R R R R
7. Primary Skin Irritation - Rabbit R R R R
8. Primary Eye Irritation - Rabbit R R R R
9. Skin Sensitization - Guinea pig R R R R
10. Cell culture (d) R R R R
11. Human Safety Records NR R NR R
(Effect/Lack of effects)
12. Toxicity to bird (1 species) NR NR NR R (Only 1
(Toxicity/Infectivity/Pathogenicity) Species)
13. Toxicity to Fresh water Fish NR NR NR R
14. Toxicity to Honey bees (e) NR NR NR R
15. Toxicity to Silkworm (f) NR NR NR R
16. Toxicity to Earthworm (g) NR NR NR R
Note:
a. Inhalation toxicity study required for registration of entomopathogenic/entomotoxic bacteria
b. Pulmonary toxicity study required for registration of antagonistic bacteria, antagonistic fungi,
entomopathogenic fungi,baculovirus
c. Intraperitoneal toxicity study required for registration of antagonistic fungi, entomopathogenic
fungi, antagonisticbacteria
d. Cell culture and Intravenous study required for registration of baculovirus. e and f – required for
all except antagonistic fungi
e. required for all except entomopathogenic/entomotoxicbacteria
Note: No data required for already registered strain from the same source with same
strain designation and accession number.
Note: If genome sequence of conserved region of the microbial strains/microbes used as
microbial pest control agent is identical with already registered strain then data is not
required from toxicity angle.
Formulations developed from similar already registered mother culture using similar
ingredient and process of manufacture then no data is required from toxicity.
Note: 1) The general recommendations as mentioned in the Guidance document on

Toxicology for registration of chemical pesticides in India will also be applicable for
registration of Microbial Pest Control Agents (MPCA) Pesticides on merit case to case
basis.
2) Waiver would be considered only when existing information provides robust and full
scientifically sound weight of evidence approach and read across/bridging from structurally and
/or biologically related similar pesticides specifically case to case basis on full merits.
3) The replacement alternatives not involving experiments on animals would not be normally
considered, except in exceptional and rare cases where in case of alternatives if available with
full and sound justification is provided specifically case to case basis on merit subject to full
satisfaction of the expert regarding full toxicity data provided for the alternatives.
PACKAGING
Chapter V of the Insecticides Rules 1971 in the Insecticides Act, 1968, the rule 16 to 20 of the
said chapter deals with the Packaging and Labelling.
Sl. Parameter Primary Formulated

No. culture/mother product
culture
9(3B) 9(3) 9(3B) 9(3)
1. Labels and Leaflets as per IR-1971, all fields (as R R R R
applicable) and as amended from time to time
2. Manner of labeling and Leaflet R R R R

3. Type of packaging R R R R
(Ultra small, small or Big whichever is applicable)
4. Manner of packaging R R R R
5. Specification for primary, Secondary and R R R R

Transport packages (whichever is applicable)
6. Details of packaging material and its compatibility R R R R

with content
7. Performance of container with content during R* R R* R
storage stability test(Shelf life Study)
8. Transport worthiness test R* R R* R

Note:
1. In case of additional packaging endorsement applications, the data at Sl. No. 05, 06,
07& 08, are not required if similar packaging (material) and manner of packaging is
being sought by the applicant as has been granted to earlier 9(3)registrant.
2. Specification of Bureau of Indian Standard (BIS) must be followed for all the packaging
requirements (Wherever available and applicable).
3. All Packaging tests must be carried out with the product of same batch and in its
commercial package preferably in Indian condition.
4. The duration of the test and the conditions including geographical conditions must be
mentioned.
5. Storage stability data must be generated keeping at least the following parameters in test
protocol viz., temperature, duration, test packaging material, content of active
ingredient in the product, test humidity, exposure to light (if applicable), physical and
chemical properties of the product during and after storage etc.
6. The testing protocols must have their basis in the WHO/FAO/ CIPAC/ASTM
recommendations or other validated methodology of GLP/ NABL accredited laboratory
having packaging testing chemical / mechanical as applicable etc.) in the scope.
7. The storage stability data for microorganisms can vary depending on the type of
microbes. For Fungi, maximum storage stability study data will be 12 months at
ambient temperature. For Gram negative bacteria like Pseudomonas fluorescens or
other Pseudomonas species the maximum storage stability study data should be 8
months at ambient temperature. For spore forming gram positive bacteria like Bacillus
species the maximum storage stability study data should be 18 months at ambient
temperature.
Note: Additional two months’ data for six months self-life claim / three months additional
data for one year and six months additional data for 18 months shelf-life claim at two/three
different agro climatic locations at ambient temperature along with meteorological data
should be submitted.
Besides this, the sub-committee also proposes the Guidelines on Consortium of Bio-
pesticides.
Guidelines on Consortium of Bio-pesticides.
Efficiency of biocontrol agents could be increased by the development of mixture of compatible
strains of different biocontrol organisms by considering the following norms. While developing
a consortia formulation, the following needs to beaddressed:
1. Compatible strains combination that differs in pattern of plant/site of colonization.
2. Compatible strains combination is broad spectrum of action against different plant
pathogens.
3. Compatible strains combination with different modes of action under similar conditions.
4. Compatible strains combination of genetically diverse group to adapt to different pH,
moisture, temperature and relative humidity.
The guidelines of Chemistry, Bio-efficacy, packaging for registration of consortia of Bio-
pesticides are similar with the guidelines of Bio-pesticides except the following points.
Guidelines of mother culture/Primary culture of already registered bio-pesticides u/s 9(3)
category are not required for registration of consortium Bio-pesticides. Only the guidelines of
formulated product (Consortium) will be required. Ratio of each strain in the formulation is
required.
1. Following toxicology guidelines for consortia of Bio –pesticides are required.
Note:
a. Inhalation toxicity study required for registration of entomopathogenic/entomotoxic bacteria
b. Pulmonary toxicity study required for registration of antagonistic bacteria, antagonistic fungi,
entomopathogenic fungi,baculovirus
c. Intraperitoneal toxicity study required for registration of antagonistic fungi, entomopathogenic
fungi, antagonisticbacteria
d. Cell culture and Intravenous study required for registration of baculovirus. e and f – required for
all except antagonistic fungi
e. Required for all except entomopathogenic/entomotoxic bacteria
Note:-
a. If genome sequence of conserved region of the microbial strains/microbes which are used in
consortia of Bio-pesticides to be used as microbial pest control agent is identical with already
registered strain, then data is not required for mother culture but data is required for
combination/consortia from toxicity angle.
b. If any new formulation of microbes is made by using new ingredients with different processes of
manufacture than data is required for the formulation.
c. If any new combination/consortia /Mixture of microbial strains/microbe developed from already
registered microbial strain than data is required only for the mixture and not for mother cultures
from toxicity angle.
List of Fungi, Bacteria and Viruses for Consortia

S. No. Name of the bio-pesticides Sr. no. in Schedule to the
IA, 1968
1. Trichoderma spp .(T. viride /T. asperellum, T. harzianum, 591
T. virensetc.)
2. Gliocladium virensT.virens 589
3. Ampyliomycesquisqualis 679
4. Coniotyriumminitans 683
5. Chaetomium globosum 692
6. Aspergillus niger(non-pathogenic/biotype) 693
7. Bacillussubtilis 588
8. Pseudomonas fluorescens, P. protegens, P.entomophila, 590
9. Streptomycesgriseoviridis 677
10. Streptomyceslidicus 678
11. Agrobacterium radiobacterK84 684
12. Beauveriabassiana 592
13. Metarrhiziumanisopliae 593
14. Nomuraea rileyi, (New name: Metarhiziumrileyi) 594
15. Verticillium lecanii, (New name: Lecanicilliumlecanii) 595
16. Verticilium chlamydosporium,(New name: 675
Pochonia chlamydosporium)
17. Paecilomyces lilacinus, (New name: 689
Purpureocilliumlilacinum)
18. Myrothecium verrucaria-nematicide 718
19. Bacillus species (includes Bacillus sphaericus (syn: 326
Lysinibacillus,Bacillus thuringiensis var. galleriae,
Bacillus thuringiensis var. israelensis, Bacillus
thuringiensis var. kurstaki, Bacillus thuringiensis var.
tenebrionsis, Bacillus thuringiensis var. sandiego,
Bacillus thuringiensis var. tolworthi and Bacillus albus.
20. Granulosis Viruses(GV) 596
21. Nuclear Polyhedrosis Viruses (NPV) (includes 597
Spodoptera litura NPV, Spodoptera frugiperda NPV,
Heicoverpa armigera NPV, Spodoptera mauritia NPV,
Mythimna separataNPV,
A. INDIAN STANDARDS ANTAGONISTIC FUNGI DRAFTSPECIFICATIONS
1. Form and appearance

2. pH
3. Composition
a. CFU/g of the product
b. Percent loading content of the Biocontrol organism in the formulation & nature of biomass.
c. Percentage of carrier/filler, wetting/ dispending agent,
d. Stabilizers/ emulsifiers, contaminants/ impurities etc.
e. Moisture content
f. CFU counts: Trichoderma 2x106CFU/ml or gm min. (Stability at 30oC and 65%RH).
4. Contaminants:
a. Biological Contaminants:
b. Pathogenic Contaminants: such as gramnegative bacteria Salmonella, Shigella, Vibrio
etc.: absent
c. Other contaminants should not exceed 1x104/ml org
d. Chemical/ botanical pesticides contaminants: absent.
5. Method of analysis:
a. CFU counts by serial dilution and examination under regular compound research
microscope with bright field optics.
b. Plating for contaminants on specific media
c. Antagonistic my colytic capability on target organism by bioassay on plants (Laboratory
test).
d. Bioassay procedure based on diseased severity and root colonization as detailed in
Appendix-I
Appendix-I
Bioassay for plant disease antagonists based on disease severity and root colonization.
The target pathogen to be tested against has to be grown in Sand maize medium. The
Sand-maize medium is prepared by adding sand 90g, maize 10g. and water 10ml in a saline or
any glass bottle of 300ml capacity and then autoclaved twice. Then 5 mycelial discs of the test
pathogen are transferred into the bottle and left for incubation for 15 days. Once the culture has
grown well, the sand maize medium is mixed along with the fungal growth and 1g from this
preparation is used as the inoculum after adjusting the CFU to 1 x10/g by addition of sand.
The plastic cups (5-6 cm diameter) filled with soil and FYM (3:1) have to be used. In
each cup the filling should be done upto ¾th level. The pathogen inoculum is mixed with sand
has to be applied upto 2cm depth in the plastic cups.
The bio-efficacy of the bio-agent shall be tested by both seed treatment and soil
application. For seed treatment, the recommended dose of the formulation has to be used (5 to
10g.). For soil application, the bio-agent is added at the rate of 1g of formulation (minimum CFU
should be the 2x106). The germination percentage, disease intensity and seedling vigour are to
be recorded.
Another set of plastic cups filled with sterile soil and sterile FYM has to be used to
confirm whether the bio-efficacy was due to the isolate of the bio-agent tested or due to the
native isolates of the bio-agent present in the soil.
The keys for grading the efficacy mentioned below shall be used (Srivastava et al.,
2002). However, for the registration purpose, the bio-agents that are Highly Efficient, Efficient
or Moderately Efficient in the plastic cup test under glass house condition (in the presence of
pathogen) can be allowed (i.e.) germination percentage of 70% or above, disease incidence of
30% or less can be considered for registration.
Disease Grading Key
Disease Description Rating of bio-

incidence efficacy of bio
(%) agents
0 Germination>90%, no seed rotting, seedling healthy, root Highly Efficient
and shoot portions well developed (HE)
1-15 Germination 80-90%, infection on main as well as lateral Efficient (E)
roots, seedlings are well developed
16-30 Germination 70-80%, development of roots restricted Moderately
and growth is less compared to Score 1. Infection Efficient (ME)
occurred on roots. Shoot portions developed but growth
retarded compared to Score 1.
31-45 Germination 60-70%, length of roots and shoots short Moderately
compared to Score 1. Germination of seeds inhibited. Inefficient
50% of root area infected. Shoot portions also showed (MI)
infection
46-60 Seed germination 50-60%. Development of roots and Efficient (I)
shoots greatly retarded. Shoot portions also showed
infection.
Above 60 Less than 50% germination and seed rotting Highly Inefficient
(HI)
For the root colonization assay, the rhizosphere region of the plants tested above have to
be collected and the soil adhering to the root surface has to be removed by gently tapping the
roots. The root bits have to be cut into 1 cm bits and randomly 25 bits should be selected for
each treatment. They have to be plated on (TSM) and the percentage of root bits colonized has to
be recorded. This has to be performed in the sterile soil and non-sterile soil. One control
treatment without the Biocontrol agent, being tested, should be kept for both the sterile and non-
sterile soil to rule out of the possibility of interference of native micro flora in the bioefficacy
assay.
B. INDIAN STANDARDS ANTAGONISTIC BACTERIA DRAFTSPECIFICATIONS
1. Form andappearance
2. pH
3. Composition
a. Percent content of the Biocontrol organism in the formulation & nature of biomass
b. CFU/g or ml of the product.
c. Percentage of other components: carrier /filler, wetting/ dispersing agent,
stabilizers/emulsifiers, contaminants/impurities etc.
d. Moisture content.
e. CFU counts: Minimum 1x108 CFU/ml or gm. (Stability at 30oC and65%RH).
4. Contaminants:
b. Pathogenic Contaminants: such as gram negative bacteria Salmonella, Shigella, Vibrio
etc.: absent
c. Other contaminants should not exceed 1x104/ml or g
d. Chemical/botanical pesticides contaminants: absent.
a. CFU counts on specific medium.
c. Antagonistic capability on target organism by bioassay.
d. Bioassay procedure based on diseased severity and root colonization as detailed in
Appendix-I
APPENDIX- I
Bio-efficacy assay for plant disease antagonists based on disease severity and root
colonization:
The pathogen to be tested against has to be grown in sand maize medium. The sand-maize
medium is prepared by adding sand 90g, maize 10g and water 10 ml in a saline or any glass
bottle of 300ml capacity and then autoclaved twice. Then 5 mycelial discs of the test pathogen
are transferred into the bottle and left for incubation for 15 days. Once the culture has grown
well, the sand maize medium is mixed along with the fungal growth and 1 g from this
preparation is used as the inoculum after adjusting the cfu to 1 x 10/g by addition of sand.
The plastic cups (5-6 cm diameter) filled with soil and FYM (3:1) have to be used. In each cup
the filling should be done upto ¾th level. The pathogen inoculum is mixed with sand has to be
applied upto 2 cm depth in the plastic cups.
The bio-efficacy of the bio-agent can be tested by both seed treatment and soil application. For
seed treatment, the recommended dose of the formulation has to be used (5 to 10g). For soil
application, the bio-agent is added at the rate of 1g of formulation (minimum cfu should be the 2
x 106, the CIB recommended dose). The germination percentage, disease intensity and seedling
vigour are to be recorded.
Another set of plastic cups filled with sterile soil and sterile FYM has to be used to confirm
whether the bio-efficacy was due to the isolate of the bio-agent tested or due to the native
isolates of the bio-agent present in the soil.
The keys for grading the efficiency mentioned below can be used here (Srivastava et al., 2002).
However, for the registration purpose, the bio-agents that are Highly Efficient, Efficient or
Moderately Efficient in the plastic cup test under glass house condition (in the presence of
pathogen) can be allowed (i.e.) germination percentage of 70% or above, disease incidence of
30% or less can be considered for registration.
Disease Grading Key
Disease Description Rating of bio-

incidence efficacy of bio-
(%) agents
0 Germination>90%, no seed rotting, seedling Highly Efficient
healthy, root and shoot portions well developed (HE)
1-15 Germination 80-90%, infection on main as well as Efficient (E)
lateral roots, seedlings are well developed
16-30 Germination 70-80%, development of roots Moderately
restricted and growth is less compared to Score I. Efficient (ME)
Infection occurred on roots. Shoot portions
developed but growth retarded compared to Score I.
31-45 Germination 60-70%, length of roots and shoots Moderately
short compared to Score I. Germination of seeds Inefficient
inhibited. 50% of root area infected. Shoot (MI)
portions also showed infection

46-60 Seed germination 50-60%. Development of roots Inefficient (I)
and shoots greatly retarded. Shoot portions also
showed infection.
Above 60 Less than 50% germination and seed rotting Highly
Inefficient (HI)
For the root colonization assay, the rhizosphere region of the plants tested above have to be
collected and the soil adhering to the root surface has to be removed by gently tapping the roots.
The root bits have to be cut into 1 cm bits and randomly 25 bits should be selected for each
treatment. They have to be plated on TSM and the percentage of root bits colonized has to be
recorded. This has to be performed in the sterile soil and not sterile soil. One control treatment
without the biocontrol agent being tested should be kept for both the sterile and non- sterile soil
to rule of the possibility of interference of native microflora in the bio-efficacy assay.
For the bacterial antagonists, the above bioassay procedure has to be followed where only the
% root colonization will be considered and other parameters are not required. The % root
colonization required is 80%.
c. INDIAN STANDARDS ENTOMOPATHOGENIC FUNGI DRAFT SPECIFICATIONS

1. Form and appearance
2. pH
3. Composition
a. CFU/g of the product
b. Percent content of the Biocontrol organism in the formulation & nature of
biomass.
c. Percentage of carrier/filler, wetting/ dispending agent
&stabilizers/ emulsifiers, contaminants/ impurities etc.
d. Moisture content
4. CFU counts: Minimum 1x108 CFU/ml or gm. (Stability at 30oC and65%RH).
5. Contaminants:
b. Pathogenic Contaminants: such as gram negative bacteria Salmonella, Shigella, Vibrio
etc: absent
c. Other contaminants should not exceed 1x104/ml org
d. Chemical/botanical pesticides contaminants: absent.
a. CFU counts by serial dilution and examination under regular compound research
microscope with bright field optics.
c. Entomopathogenic capability on target insects by bioassay.
Appendix-I
Laboratory bioassay procedures for screening fungal pathogens on Spodoptera litura and
Helicoverpa armigera
Insect pathogens:
Beauveria bassiana, Metarhizium anisopliae, Nomuraea rileyi
Preparation of Fungal inoculum for bioassays:
The fungus is grown on SDAY/SMAY medium for 10 days in slants and aqueous spore
suspensions of various concentrations are prepared using sterile water. The spore count is
estimated by Haemocytometer. (104 - 1010 spores/ml). Tween-80 is added @ 0.01% to get
uniform spore suspension.
Rearing insects:
H.armigera,S.litura - Artificial diet(Semi-syntheticdiet)
Stage of insect for bioassay
H.armigera, S.litura - II instar larvae to be used for bioassay protocols for lepidopteron pests
Method of inoculation
S. litura
1. Cut castor leaf discs of 3.0cm diameter, rinse in sterile distilled water and place each leaf
disc in a sterile Petri plate and allow it air dry in a laminar flow system
2.Apply ten micro liters of the spore suspension of each concentration on the leaf disc and
spread it uniformly on the leaf surface and allow it air dry in a laminar flow system. Treat the
other side of the disc similarly.
3. Release ten numbers of second instar larvae of S. litura on the leaf surface and incubate the
discs in an incubator at 250C and 90%RH
4. After 24hours, shift the larvae to the polypots containing the semi-synthetic diet and incubate
in an incubator at 250C and 90%RH
5. After 5 days of incubation, mortality of the larvae are recorded in each concentration tested
6. Lc-50 can be calculated using SPSS package
Standard for LC-50: Not more than 2.00X106 spores/ml (3.0X103 spores/mm2)
H. amigera:
Instead of castor leaves, soybean leaves can be used for H. amigera and the procedure is same as
above.
Standard for LC50: Not more than 4.00X106 spores/ml (6.0X103 spores/mm2)
Appendix-II
Bioassay procedure for Plutella xylostella
Various concentrations of Beauveria bassiana formulation ranging from 6 x 108 to 2 x 1010 are
to be screened to assess the mortality.
Fresh undamaged radish leaves free from pesticide application are to be collected and washed
thoroughly in sterile distilled water and air-dried. Individual leaves are dipped in respective
concentrations for 30 seconds. After complete drying of leaves ten late 2 nd instar larvae of
Plutella xylostella are released per treatment. A water dipped radish leaf is maintained
simultaneously as control.
To prevent desiccation of leaves, the petiole is covered with a moist cotton swab. Each treated
leaves are placed in a plastic container of dimension 12.5 x 10 cm containing moist filter paper,
Whatman No.41 to provide humidity.
Each treatment has to be replicated thrice. Fresh radish leaves were provided as feed at 24 hours
interval. This set up has to be maintained at 25+10C and 70-80% RH for 7 days. Observations
on larval mortality are to be made at 3, 5 and 7 days after treatment.
Standard for LC50 = Not more than 3 x 109 cfu/g

D. INDIAN STANDARDS, ENTOMOTOXIC BACTERIATECHNICAL /FORMULATION

DRAFT SPECIFICATIONS
S.No. Details
1.SCOPE
1.1 This Indian Standard prescribes the requirements and the method of sampling and
test for Entomotoxic bacteria technical and formulation. The product is a bio-
pesticide active against target insects. The product is not for human consumption.
2.REQUIREMENTS
2.1 Common name: i.e., Bacillus thuringiensis or B. sphaericus etc.
2.2 Systematic name (Genus, species, serotype, strain and Cry-toxin* along with cry
gene)
2.3 Physical specification
2.3.1 Form and appearance

2.3.2 Moisture content
2.3.3 pH
2.4 Composition
2.4.1 Delta endotoxin content (Minimum 2.0% ) – estimation as per
Appendix-V
2.4.2 Adjuvants
2.4.3 Beta Exotoin content – Negative through housefly bioassay test as per
Appendix-IV
2.4.4 Human pathogens (gram negative bacateria Salmonella, shigella &vibrio
etc) -Absent
2.4.5 Other microorganisms (not more than 104 /g)
2.4.6 Chemical/botanical pesticide contamination –Absent
2.5 Natural occurrence of the organism
2.5.1 Its relationship of the organisms
2.5.2 History (exotic or indigenous strain)
2.5.3 The isolate should not be genetically modified organism(GMO).
3.SAMPLING
3.1 Representative samples of the material shall be drawn in accordance with IS
10946:1984
4.TESTS
4.1 An appropriate test procedure and criteria used for identification, such as
morphology, biochemistry and / or serology / immunology
4.1.1 Morphology description, particle size

4.1.2 Immunology assays: ELISA / Dot blot assay test or any other sensitive
standard immunology test.
4.1.3 Method of analysis
4.1.4 Level of beta exotoxins to be identified if expressed by Housefly
bioassay method.
4.1.5 Potency of product by bioassay method(Appendix-II)
4.1.5.1 Bioassay method
a) LC 50 on target larvae and potency against a reference
using artificial diet or leaf disc method or in the water
for mosquito larvae(Appendix-I)
b) Housefly Bioassay method for Beta-exotoxin content
(Appendix-IV)
c) Determination of toxin content by ELISA / Dot Blot
Assay Method(Appendix-V)
4.1.5.2 A technique for the separation and purification of the crystals
(Appendix III) is to be used by the manufacturer and the antisera to be raised
using solubilized toxin. Toxin content (3.5 %) to be standardized in the
formulation using this antisera (ELISA /Dot blotassay).
2.2 Crytoxin* If H-Serotype is not known, it is mandatory to provide the details of Cry
toxin to confirm that it is Bacillus thuringiensis.
Appendix I
Bioassay Method
Diet incorporation
The following protocol is used for diet incorporation of oral toxicants to test their toxicity on
target insects. The example presented here is to bioassay Cry I Ac on H. Armigera (First instar
larva of other test insects are used for similar bioassay).
1. Pipette out 3 ml of the solution into a 40 ml plastic cup.

2. Pour lukewarm diet, approx. 600 C, into the cup to a total volume of 30 ml. Place the lid
and shake the cup vigorously for a minute to mix properly.
3. Pour the diet to 0.5 cm height, into wells of a 24-cell insect- rearing tray. Allow the diet
to cool in laminar airflow under UV lamps for I h to surface sterilize the diet.
4. If concentration of the toxicant in the stock solution was 2 µg/ml, the final concentration
in the diet would now be 0.2µg/ml diet. Thus the final concentration of toxin in diet was
diluted10-fold.
5. Release first instars into the diet rearing trays at the rate of one per well. Cover the diet
tray with semi-permeable wrap and close the lid.
6. It is recommended that the lid be tightly secured to the tray with rubber bands, to prevent
the larvae from escaping. Because the diet is unsuitable, larvae try constantly to escape
from the diet rearing trays.
7. Keep controls with larvae released on untreated diet, for all the experiments.
8. The unused rearing trays with diet can be stored in a refrigerator for a week.
9. Change the diet for the larvae every two or three days.
10. Record mortality observations at 8 hourly intervals until the end of seven days, for
median lethal time LT50 calculations. LT50 is the time at which 50 % of' the test
population is killed with the specific dose tested. A simple linear regression equation can
be worked out to calculate the LT50
11. Otherwise, record mortality at alternate days until the end of seven days, for median
lethal concentration LC50 calculations. LC50 is the concentration that kills half the test
population.
12. Record weights of surviving larvae at the end of seven days, for median effective
concentration EC50 and LC50 is the concentration that prevents half the test population
from reaching 50% of the weight attained by control larvae. For example, if the average
weight of larvae on the control diet (without toxin) was 80 mg, EC50 represents the
concentration at which 50% of the test population is unable to gain a weight more than 40
mg. LC50 is the concentration that inhibits half the test population from reaching the third
instar.
Diet incorporation for filter paper bioassays

1. For bioassays with bollworms, 10 ml toxin incorporated diet is poured over a16 sq cm
piece of filter paper. The filter papers layered with diet are cooled and cut into smaller
squares of 2 x 2 cm, and 10 first instar larvae are released in small plastic cups 3 x 3 cm
(d x h) cups containing a square. Change the strips every alternate day.
2. Record mortality observations until the seventh day.

Surface coating of semi-synthetic diet

1. Prepare the diet and pour it into the trays or the rearing plastic cups. Generally, 10 µl of the toxin
can be used to coal 1 sq cm surface area. Gently swirl the diet surface to ensure uniform and
complete spread of the solution over the diet surface.
2. Allow the surface to dry in a laminar airflow under UV light for 2-3 hours to surface sterilize.
3. Release one first instar H. armigera larva per well. Always maintain proper controls with
untreated diet.
4. Change the diet on alternate days and record mortality until the seventh day. Then, weight of
surviving larvae should be recorded on the final day of the bioassay.
The method has the advantage of obtaining constantly reliable results because the toxin is
unlikely to be affected by either improper mixing or heat as can occur in the diet-incorporation
method. Moreover, less amount of the toxin is required for the assay, compared to the diet-
incorporation method.
Calculation of results:
The potency of the sample (International Units – IUs)
LC50 Standard
IU/mg sample = -------------------- X IU/mg Standard
LC50 Sample
(IU/mg Standard, i.e., HD-1-S-1980 is 16,000 IUs/mg; the US standard is available with PDBC,
Bangalore; each registrant should prepare a “self reference” and should deposit it with the
Registering Authority. Each self reference will be expressed as IU/mg using International
standard)
Exotoxin determination by PCR studies
Methodology:
Some Bacillus thuringiensis strains secrete type I β- exotoxin, which is a non-specific insecticidal
and thermostable adenine nucleoside oligosaccharide. Toxicity bioassays and HPLC are
traditional methods for detecting β-exotoxin. For rapid approach for prediction of type I β-
exotoxin production, PCR-based method can be followed as per Diego H Sauko et al. (2014). One
of these ORFs encodes the Exo protein that was proved to be responsible for the phosphorylation
of a β-exotoxin precursor at the last step of their biosynthesis process (Liu et al., 2010). Primers
BEF (forward; 50- CGGCAGCCGTTTATTCAAA-30) and BER (reverse; 50-
CCCCTTCCCATGGAGAAACA-30) amplify a 406-bp DNA fragment of thuE between
nucleotides 373 and 778. All B. thuringiensis strains are grown on nutrient agar plates for 16h. A
loopful of cells is transferred to 100 µl H2O and boiled for 10 min to make DNA accessible for
PCR amplification. The lysate is centrifuged briefly (5 s at 20,000g), and 5 µl supernatant is used
as a DNA template in each polymerase chain reaction. This is performed with a final volume of
25 µl containing 2.5 µl 10x reaction buffer, 0.5 µl 50 mM MgCl2, 0.5 µl 100 mM
deoxynucleoside triphosphate mixture, 8 pmol each primer, and 1 U of Taq polymerase
(Invitrogen). The PCR amplification consisted of DNA denaturation at 94°C for 2 min followed
by 25 cycles of amplification with a gradient thermocycler. Each cycle consisted of a denaturation
step at 94°C for 1 min, an annealing step at 54°C for 1 min, and a chain elongation step at 72°C
for 1 min. The final elongation step was extended for an additional 5 min. Subsequently, 10 µl
PCR product is analysed by 1.0% agarose gel electrophoresis. A positive control can be also used
for better results.
75
Appendix-II
Dot Blot assay of Bacillus thuringiensis (B.t.) toxin protein as alternate of Bioassay.
1) B.t. grown till sporulation in shake flask or in fermenter vessel and let the cells lyse and
release spore/crystals into the medium
2) Cells are harvested by centrifugation at 10k for 15mins.
3) Wash the pellet with 1M NaCl to remove the B.t. associated seine/metallo proteases and
washed twice with sterile distilled water.
4) Pellet suspended in 50MM NaOH to solublize the toxin protein for 2 hours at R.T. with
slow shaking and centrifuged again at 10K for 15Mins.
5) Supernatant was adjusted to pH 8.0 with Tris HCL pH8.8
6) Protein contents estimated by Lowry’ sprotocol.
7) Two fold serial dilutions of test protein were made in PBS and known amount at
protein applied on NCP using S&S or Biorad Dot Blot manifold apparatus and applying
water vacuum for 30mins.
8) NCP was carefully removed from Dot Blot set and soaked in excess of 3% Skim milk in
PBS for blocking the remaining acetic sites on NCP for 2-3 hours at R.T/O/N at4oC.
9) Wash the NCP with excess PBS with 0.01% Tween 20, 3-4 times and then finally with
PBS
10) Polyclonal antiserum raised against total crystal protein was suitably diluted in PBS and
added to the ‘seal a meal’ containing NCP and incubated for 1-2 hours with shaking.
11) Remove the NCP from the bag and was several times (as mentioned instep.No.9)
12) Anti-rabbit antibodies conjugated with HRPO/alkaline Phosphate was diluted as per the
suppliers instruction and incubated NCP (as in step10)
13) Was as in step11
14) For HRPO:
a) Diaminobenzene (4mg/10ml PBS)/4-Chloro-1-Napthol (4mg/10ml 20%

Alcohol) were dissolved and 10ml of 30% of H2O2 per 10 ul substrate solution
was added and colour reaction developed in dark for 5-10 mins (DAB gives
brick red colour. 40N gives bluecolour).
b) For alkaline Phosphatase:
Alkaline Phosphatase Buffer:
1M TrispH8.8 -10ml/
4MNaCI - 2.5ml/ make up to100ml
1MMgc12 -0.5ml/
For 10ml of above buffer add NBT-66 ul and BCIP-33 ul and developed and colour reaction
15. Stop the reaction by removing the substrate and washing with PBS.
16. Keep on filter paper and dry.
DIFFERENT PROTEIN CONCENTRATION
10ug 5ug 2.5ug 1.25ug 512.5ng 256.25ng 128ng 64ng 32ng 16ng 8ng 4ng
Determination if cell dry weight
# Take a known volume of Bacterial culture spin down at 4R form in.

# Wash the pellet in minimal distilled water
# Transfer to a pre weighed container
# Incubate at 800C for 16-18 hours till become dry and weight become sconstant.
Appendix-III
PURIFICATION OF CRYSTALS BY GELATIN METHOD
Centrifuge the sporulated material and wash pallet twice with 1M Nacl. Add 200ml. of
0.5% Gelatin, stir and remove all froth completely. Dilute with sterile water and centrifuge. Take
debris and stir with 20ml. of 1.5M sucrose. Further add 50 ml of 1.5M sucrose, stir and centrifuge
at 3000 RPM for 2 hours. Remove supernatant and purified crystals are harvested.
E. INDIAN STANDARDS BACULORIVUS DRAFTSPECIFICATIONS
1.Form and composition of the product

1.1 Viral Unit: POB/Capsule count per ml/g of the product
1.2 Percent content of the bio-control organism in the formulation and nature of biomass
1.3 Percent of carrier/filler, wetting/dispersing agent, stabilizers/ emulsifiers, containments/
impurities etc.
2. Moisture content
3. pH
4. Viral Unit:
NPVs (Helicoverpa & Spodoptera) - 1x109 POB/ml or gm (minimum) (POB –Polyhedral

Occlusion Body)
GV (Chilo, Plutella & Acheae) - 5x109 Capsules/ml or g. (minimum).

5. Contaminants:
5.1 Biological contaminants:
5.1.1 Pathogenic contaminants: Pathogenic contaminants such as gram negative bacteria
Salmonella, Shigella, Vibrio etc. should be absent:
5.1.2 Other microbial contaminants: Other microbial contaminants should not exceed 1x10 4
/ml or g
5.2 Chemical/botanical pesticides contaminants should be absent.

6. Identification of Baculovirus by DNA test (Restriction enzyme analysis test).
7. An undertaking should be submitted that the strain is indigenous, naturally occurring and not
exotic and not genetically modified as perAnnexure-1.1
Viral Unit:
NPVs (Helicoverpa and Spodoptera) =1x109POB/mlor gm. minimum
GVs = 5x109 Capsules/ml or gm. minimum.

8.1 In case of NPVs/, POB/Capsule count should be taken with Haemocytometer using
shallow depth counting chamber as detailed in Appendix – I
8.2 Biological assay for determining the LC50 or LD50 of the formulation:
8.2.1 Bioassay for NPV by the Diet Surface Contamination Method as detailed in Appendix-II
OR
8.2.2 Bioassay for GV against Chilo infuscatellus asdetailedin Appendix-IIIOR
8.2.3 Bioassay for GV against Plutella xylostella as detailed in Appendix-IV.
8.2.4 Bioassay for GV against Acheae janta as detailed in Appendix-V.
8.3 Plating for contaminants on specified media.
Appendix-I
COUNTING OF NPV/GV (POB/CAPSULE) USING IMPROVED NEUBAUER

HAEMOCYTOMETER COUNTINGCHAMBER.
A haemocytometer is used for estimating of NPVs/GVs in a unit volume of the product.

The Improved Neubauer Haemocytometer comprised a thick glass slide with a shallow
depression in the central section divided into two halves (figure-1). Each side, the base of the
depression has a fine ruled grid of squares (figure-2) which is visible under a microscope. The
dimensions of this grid are defined. Place a standard cover slip placed over the depression and a
one half halves of the slide chamber using a micro pipette. The particles require 2-5 minutes to
sediment to the chamber floor.
Either dark field or a phase contrast microscope is used to identify and count polyhedral
occlusion bodies (POB) or capsule. With the counting chamber under the microscope, the
number of Polyhedra/capsule in a given number of grid squares can be counted. Each count
consists of a tally of the number of polyhedra completely contained within a big square plus the
number of touching the top and left sides. Polyhedra touching the bottom and right sides are not
counted. Since both the depth of the chamber and the grid dimensions are known. It is then a
straight forward calculation to determine the number of polyhedra /capsule per ml of test
suspension.
Number of NPV (POB) per ml/gm = D x X

NxK
Where:
D = Dilution factor
X = Total number of polyhedra
counted N = Number of squares
counted
K = Volume above one small square in cm3 =(2.5x10-7cm3)
Area of each small square is 1/400 mm2 = 0.0025 mm2. Depth of chamber is o.1mm. Volume of
liquid above a single small square is 0.0025 mm2 x 0.1mm= 0.00025 mm3. To covert to cm3
multiply by 1/1000 to get a volume of 2.5 x 10-7 cm3 above 1 small square. Hence,K=2.5x10-
7cm3
Worked example:
Suppose in a sample diluted by a factor of 1000 we count 535 polyhedra in 160 small squares
then:
D = 1000
X = 535
N = 160
K = 2.5 x 10-7 cm3
1000x535
Thus, POB count=-------------- =1.34x1010POB/ml of test sample
160x2.5x10-7
Note: (i) Usually,thisprocedureisrepeated3timesandanaveragetakentogetamore accurateestimate.

(ii)Same procedure will be used for GV also for counting the number of capsuleper unit
volume of the product.
Appendix-II
PROCEDURE FOR ESTIMATION OF LC50 OF NPV BY THE DIET SURFACE

CONTAMINATION METHOD.
i) Diet to be used: The standard chickpea-based diet without formaline.

ii) Bioassay bottles: 5ml. vials with a diameter of 18 mm (255 mm2 surface area)
iii) Doses of NPV to be tested:
Helicoverpa armigera Spodoptera litura
POB/ml POB/mm2 POB/mlPOB/mm2

--------- ---------- ------- -------
a)5x104 1.96 1x106 39.21

b)1x104 0.39 2x105 7.84
c) 2x103 0.078 4x104 1.57
d)4x102 0.016 8x103 0.31
e)0.8x102 0.003 16x102 0.062

f)1.6x10 0.0006 3.2x102 0.013
iv) Method of dosing: Dispense 10 Microlitre aliquots into each vial and spread
uniformly over the entire diet surface using a polished rounded lip of 4 mm glass
rod and allow to dry off under flow laminar hood for 10minutes.
v) No. of larvae/dose:50 (Maintain 50 healthy larvae without virus
inoculation for control)
vi) Stages of larvae: II instar larvae (Preferably 4daysold) Release one larva/vial
and plug mouth with sterile absorbent cotton. Incubate at 25 ± 1oC for 7days.
vii) Record mortality in different doses on the 7thday.
viii) Apply Abott’s formula for correction of mortality in control treatment.
ix) Subject the dose – mortality response to probit analysis using relevant statistical
software.
x) Express LC 50 as POB/mm2 of diet surface.
------------------------------------------------------------
Expected standards for NPV for II instar larvae
------------------------------------------------------------
Species LC 50POB//mm2
1. Heliocoverpaarmigera <0.5
2. Spodoptera litura <20.0
Appendix-III
Bioassay for GV against Chilo infuscatellus
Determination of LD50:
To determine the LD50 of the GVs, third instar larvae should be used. The larvae are to be
microfed (one micro litre per larva) with six different doses, viz. 1.1 x 101, 102, 103, 104,
105, and 106 IBs/larva. One hundred freshly moulted larvae have to be used for each
treatment. Larvae fed with equal quantity of distilled water serve as control. The mortality
has to be recorded daily. The LD50 of the virus is determined following the probit analysis
method (Finney,1962).
LD50 = < 1x103 OB for third instar larvae by micro-feeding.

Appendix-IV
Laboratory bioassay procedures for estimation of LC50 of Plutella xylostella (PxGV) by

leaf disc method:
1. Cut leaf discs of cauliflower (3.2cm). Soak it in 0.1N NaOCI for 5 min. and wash
thoroughly in distilled water. Air dry these leaf discs for 2-3 minutes. (Fifth leaf from
top to be used)
2. PxGV (containing 0.01per cent Triton X 100) of different concentrations 28000, 2800,
280, 28, 2.8 OB/mm2 on the leaf disc) is prepared

3. Aliquots of 12ul of each concentrating of GV is dispensed on the upper surface of the
leaf disc and spread uniformly with a blunt end glass rod (use separate tips and glass
rods for each treatment)
4. Air dry these leaf discs for 2-3minutes
5. Repeat the same on the lower surface of the leaf disc.
6. Control discs were treated with distilled water containing 0.01 per cent Triton X 100
only.
7. The leaf discs are placed in Petri dishes lined with wet filter paper discs and 35-second
instar larvae of P. xylostella (starved for 6 hours) are released on each leaf disc starting
from control treatment to highest concentration. This is replicated three times.
8. Incubate these larvae at25oC
9. After 24 hours remove the treated leaves (partially eaten) and provide the larvae with
fresh cauliflower leaves.
10. The leaves are changed daily and mortality data recorded everyday.
11. The dosage and time mortality responses are subjected to probit analysis.
12. If the mortality in the control excess 10% repeat the experiment.
LC50 = < 0.15 OB/mm2 for second instar larvae by disc method.
Appendix-V
Laboratory bioassay procedures for estimation of LC50 of Achaea janata Granulosis virus
(AjGV) by leaf disc method:
1. Cut leaf discs of castor (8cm dia) and wash in distilled water. Air dry these leaf discs
for 5minutes.
2. Treat the leaf disc on both the upper and lower surfaces with 200 μl suspension of
AjGV (containing 0.02% Tween-80) of different concentrations (5x108, 5x107 , 5x106
, 5x105 , 5x104 corresponding to 19884, 1988, 198, 19, 1.9 OB per mm2 on the leaf
disc)
3. Aliquots of 100 μl of each concentration of GV is first dispensed on the upper surface
of the leaf disc and spread uniformly with a blunt end of glass rod (use separate tips
and glass rods for each treatment)
4. Air dry these leaf discs for 5minutes
5. Repeat the same on the lower surface of leaf disc
6. Control leaf discs were treated with distilled water containing 0.02% Tween-80only
7. The leaf discs are placed in Petri dishes (9.0cm dia) line on wet filter paper discs and
35 second instar larvae (third day after hatching) of A. janata are released on each leaf
disc starting from control treatment to highest concentration. This is replicated three
times.
8. Incubate these larvae at25oC.
9. After 24-48 hours remove the treated leaves (partially eaten) and provide the larvae
with fresh castor leaves
10. The leaves are change daily and mortality data recorded everyday
11. The dosage and time mortality responses are subjected to probit analysis
12. If the mortality in the control exceeds 10% repeat the experiment.
Recommended LC50 GV (Achaea janata) – LC50 <4 OB/mm2 for second instar larvae by
the leaf disc method.
ANNEXURE IV
The Data Requirements for Botanical/ Plant origin Pesticides .
II: Plant Based Products
Botanical Pesticides/Pesticides of any Plant Origin like Eucalyptol, Cymbopogon

formulation, Neem Based Pesticides, Pyrethrum Extract & Rotenone for Pisci-culture
(Formulation)] & Essential Oils (formulations only).
General Guidance
1. It is clarified that traditional remedies or products prepared by the individuals for

their own-selfuse do not need registration if the produce is not sold. If the product
prepared or the treated produce is to be sold in the market, then the registration is
required under the provisions of the Act.
2. Sometimes the active substances to be used as Plant origin or botanical bio-
pesticides, are vastly studied and the information of good quality which is consistent
with current methodology, information/data is available, then waivers for submission
of certain data, if justified, can be granted. Further, in some cases, based on the
information and characteristics or the results of studies, additional data may be
required.
3. For registration of extract u/s 9(4): It shall be obtained from the same part of the plant
with same process of extraction and should have identical chemical composition as of
9(3) registrant.
4. Formulation to be registered u/s 9(4) should have identical chemical composition and
required to be prepared from the extract already registered under the Act for use in
the country.
Chemistry
Plant extract like Eucalyptol, Cymbopogon, Neem Based Pesticides, Concentrate Pyrethrum
Extract, Herbal/ Botanical plant growth regulator , Rotenone for Pisci-culture and Plant
essential oil etc.
S. Parameter Botanical
No Plant extract/ Formulation

concentrate
9(3b) 9(3) 9(4) 9(3b) 9(3) 9(4)
1 Name of the Part of the Plant(s) R R R R R R

to be used for extraction of the
active ingredients.
2 Source of supply of the technical R R R R R R

grade material
3 Outline of Process of R R R R R NR
Formulation.
4 Extract Contents active ingredient NR NR R R R R
5 Chemical composition of the R R R R R R

formulations
6 Physico-chemical properties of R*/ R*/ NR NR R*/ NR R*/ NR NR

a.i. & adjuvants NR
7 Product Specifications as BIS R R R R R NR

format
8 Method of analysis (a.i.) R R R R R NR
9 An undertaking that the product R R R R R R

does not contain any other
chemical pesticide except plant
extract
10 Analytical test report R R R R R NR
11 Shelf-life claim R R R R R R
12 Shelf-life data NR R NR NR R NR
1. R*: New Molecule which is being introduced for first time into the country.
2. Shelf life data: It should be generated at three agro-climatic zone.
Bio-efficacy
Parameters Technical Formulations

9(3B) 9(3) 9(4) 9(3B) 9(3) 9(4)
1 Bio-effectiveness R R No bio- R R No bio-
against target pest (Two (Two season efficacy (Two season (Two efficacy
species in specified season data from data data from season data
crops data from min. Three require min. Two data from required.
min. Two agro d. agro min.
agro climatic climatic Three Claim
climatic zones) Certific zones) agro will be
zones) ate of climatic granted
Registr zones) as per
2 Phytotoxicity (as R R ation R R approve
per standard tests) (same as (same as will be (same as (same as d
above 1) above 1) granted above 1) above 1) formulat
3 Compatibility with R R as per R R ion u/s
other agro- approve 9(3).
chemicals, if d
claimed technic
4 Stability of R R al u/s R R
formulation (Photo, 9(3)
Thermal etc.) in
aqueous dilution
(acidic, neutral &
basic)
5 Direction R R R R
concerning dosage
for each target pest
species
6 Stage of crop for R R R R
use and stage of
target pest
7 Waiting period R R R R
8 Application R R R R
equipment
9 Information R R R R
regarding
registration status
in other countries,
if any.
10 Time of R R R R
Application
11 Purpose of R R R R
Manufacture
13 Residue in Plant R R R R
14 Residue in Soli R R R R
15 Registration status R R R R
in foreign countries
16 Residue tolerance R R R R
limits fixed by
foreign countries)
R – Required NR – Not Required.
Note :
1. Sr. No. 4 & 6 not required in case of Cymbopogan formulation (Plant extract) & Neem Based
Pesticides
2. Sr. No. 1,2,3,,5,6,7,8,9,10 applicable for Essential oil for Formulation only.
Toxicology
Parameters Extract/concentrate Formulation

9(3b) 9(3) 9(4) 9(3b) 9(3) 9(4)
1. Acute oral toxicity – Rat R R No data R R No
2. R R required if R R data
Acute dermal toxicity –
chemical require
Rabbit
equivalence d if
3. Acute Inhalation - rat R R is R R chemi
4. Primary Skin Irritation – R R established R R cal
Rabbit with 9(3) equiva
5. Primary Eye Irritation – R R registrant. R R lence
Rabbit is
6. Skin Sensitization - R R R R establi
Guinea pig shed
7. NR NR/R* NR NR with
Sub- acute oral – rat
9(3)
8. NR R NR NR registr
Sub - acute oral – dog**
ant.
9. NR R NR NR
Sub- acute dermal
10. NR R NR NR
Sub- acute inhalation
11. NR R NR NR
Neuro-behavioral
toxicity
12. NR R NR NR
Teratogenicity
13. NR R NR NR
Effect on Reproduction
14. NR R NR NR
Carcinogenicity/chronic
toxicity/combined
toxicity study) *
15. NR R NR NR
Metabolism
16. NR R NR NR
Mutagenicity* (AMES +
2 in-vitro +1 in-vivo)
17. Toxicity to birds R (Two R (Two R (one R (one
species) species) species) specie
s)
18. Toxicity to fresh water R R R R
fish
19. R R R R
Toxicity to Honey bees
20. R R R R
Toxicity to Earthworm
21. R R R R
Medical data
22. NR/R* NR/R* NR/R* NR/R
Human toxicity
*
information
23. NR R NR R
Health record of
industrial workers
24. NR/R NR/R NR/R NR/R
International report on
carcinogenicity &
genotoxicity study
Note:
1. The requirement for sub-acute studies shall be determined on the basis of results of other
toxicity study reports.
2. *PLEASE REFER TO GUIDANCE DOCUMENT ON TOXICOLOGY FOR

REGISTRATION OF CHEMICAL PESTICIDES IN INDIA.
Toxicity study No.8-Sub acute oral toxicity study (Dog): **Peer reviewed international
published literature is also acceptable for dog study.
registration of botanical pesticides on merit case to case basis.
scientifically sound weight of evidence approach and read across/bridging from structurally
and /or biologically related similar pesticides specifically case to case basis on full merits.
3) The replacement alternatives not involving experiments on animals would not be
normally considered, except in exceptional and rare cases where in case of alternatives if
available with full and sound justification is provided specifically case to case basis on
merit subject to full satisfaction of the expert regarding full toxicity data provided for the
alternatives.
PACKAGING
Chapter V of the Insecticides Rules 1971 in the Insecticides Act, 1968, the rule 16 to 20 of the
said chapter deals with the Packaging and Labeling.
Sl. Parameter Extract/concentrate Formulation

No.
9(3B) 9(3) 9(4) 9(3B) 9(3) 9(4)
1. Labels and Leaflets per IR- R R NR R R NR
1971, all fields (as
applicable) and as amended
from time to time
2. Manner of labeling and R R NR R R NR
Leaflet
3. Type of packaging R R NR R R NR
(Ultra small, small or Big
whichever is applicable)
4. Manner of packaging R R NR R R NR
5. Specification for primary, R R NR R R NR

Secondary and Transport
packages (whichever is
applicable)
6. Details of packaging R R NR R R NR
material and its
compatibility with content
7. Performance of container R* R NR R* R NR
with content during storage
stability test(Shelf life
Study)
8. Transport worthiness test R* R NR R* R NR
Note:
1. In case of additional packaging endorsement applications, the data at Sl. No. 05, 06, 07& 08, are
not required if similar packaging (material) and manner of packaging is being sought by the
applicant as has been granted to earlier 9(3) registrant.
3. All Packaging tests must be carried out with the product of same batch and in its commercial
package preferably in Indian condition.
5. Storage stability data should be generated in consonance of BIS or product specification.
6. The testing protocols must have their basis in the WHO/FAO/ CIPAC/ASTM recommendations
or other validated methodology of GLP/ NABL accredited laboratory having packaging testing
(chemical / mechanical as applicable etc.) in the scope.
7. The Accelerated storage study (ASS) test must be conducted at 54± 2oC (wherever applicable) for
14 days as per FAO/ WHO manual for claiming appropriate shelf life of the product which can be
maximum two years, subject to the condition of providing the ambient storage stability study data
of thirty months or as the case may be within thirty months from the date of application for the
registration.
***
ANNEXURE V
THE DATA REQUIREMENTS FOR PHEROMONES/SEMIO-CHEMICALS
The Data Requirements for Pheromones/Semio-chemicals Chemistry
Technical for import/indigenous manufacture:
S Parameters 9 (3b) 9(3) 9(4)

No
1. Active ingredient/Tech conc. R R R
2. Chemical composition R R R
3. Chemical name (s) R R R
4. Common name R R R
5. Physical chemical properties R R R
6. Manufacturing process R R R
7. Analytical Test Report from R R R
GLP/ NABL accredited
laboratory
8. Storage condition with special R R R
reference to temperature
9. Source of import ( In case of R R R
import only)
10. Sample for PRV purpose R R R
11. Purpose of import/indigenous R R R
manufacture
1. TECHNICAL FOR IMPORT/INDIGENOUS MANUFACTURE:
1. Active ingredient
2. Laboratory test: Lure manufactured from the particular ingredient will be tested by using
wind tunnel
and should demonstrate minimum 50% attractancy.
LURE/DISPENSER FOR IMPORT/MANUFACTURE:
1. Laboratory test; The lure/dispenser should demonstrate at least 50% attractacy using
the wind tunnel.
2. Field test: The data on bio-efficacy based on two seasons field trials from two
different agro-climatic
conditions in the form of authentic/published report.
3. Compatibility: No data on compatibility are required unless the product is

recommended for use in
combination with pesticides or other agrochemicals.
4. Time and method of application: Information on timing, disruption is to be furnished.

5. Intended uses.
6. Mode of action and degree of specificity.
7. Target pest (s) and crops or premises to be protected.
8. Application rate.
9. Manner, rate and frequency of application.
10. Limitations of use.
Note: As per decision of 248th meeting of R.C. Pheromones used for monitoring and
mass trapping are not covered under the various provisions of the Insecticides Act,
1968.
Bio-efficacy
S. Particulars 9 (3b) 9(3) 9(4)

No.
1.
Active ingredients R R No bio-
Laboratory test (attractancy) : R
2. R efficacy data
Lure manufactured from the required.
particular ingredient will be
tested by using wind tunnel Claim will be
and should demonstrate granted as per
minimum 50% attractancy approved
LURE/DISPENSER FOR IMPORT/MANUFACTURE: formulation
3. Laboratory test: The R R u/s 9(3).
lure/dispenser should
demonstrate at least 50%
attractacy using the wind
tunnel.
4. Field test The data on bio- The data on

efficacy based on bio-efficacy
two seasons field based on two
trials from one agro- seasons field
climatic trials from
conditions in the two different
form of agro-climatic
authentic/published conditions in
report. the form of

authentic/pub
lished report.
5. Compatibility: No data on R R
compatibility are required
unless the product is
recommended for use in
combination with pesticides or
other agrochemicals.
6. Time and method of R R
application: Information on
timing, disruption is to be
furnished.
7. Intended uses. R R
8. Mode of action and degree of R R
specificity.
9. Target pest (s) and crops or R R
premises to be protected.
10. Application rate. R R
11. Manner, rate and frequency of R R
application.
12. Limitations of use. R R
Note: As per decision of 248th meeting of R.C. Pheromones used for monitoring and
mass
trapping are not covered under the various provisions of the Insecticides Act,
1968.
SPECIFICATIONS FOR PHEROMONE TRAPS AND LURES
The committee after detailed discussions on the existing specification of various models of
pheromones traps decided not to make any change in these specifications. However, these
specifications may further be examined by SAU and SDAS of various States:
1.Funnel shaped trap: This trap is generally used for trapping the moths of Hellcovarpa
armigera,
Spodoptera litura, Earisa spp. etc.
Colour: Any colour other than black.
Structure: The funnel trap may have three parts (1) canopy (2) funnel shapped “trap base”and
(3) a
collection device.
Canopy: Dia : 120-160mm
Thickness : 1.0-3.0mm
(There should be a provision for fixing the canopy to the “trap base” and also the (pheromone
lure)
Trap base:
Dia of the mouth : 75-120mm

Height of funnel : 45-190mm
Dia of the bottom hole : 20-30 mm
Should possess a “L” or “T” shaped handle or any other device by which the other device by
which the “trap” may be fixed to the support.
The “Trap base” may be provided with 2 to 4 stalks for fixing the canopy to the “trap base”. The
canopy should be firmly rest on stalks so that the canopy is not dialodged due to wind.
Collection device: It should be made of polythene or other suitable material. It should withstand
wind,
temperature and rain water.
Should be fixed to the“trap base” in such a way that the device remains attached to the trap under
field conditions.
II. Sticky trap (for pink boll worm etc.):
* Corrugated DVC, Plastic laminated card board, tin or any other suitable material that
should be water-proof.
* The sticky glue should be non- drying.
* The outer surface of trap should be water proof.
* The colour may be except black.
* There should be provision for fixing the trap for support.
III. Fly trap (For fruit/vegetable flies):
* Material construction as described in sticky/funnel trap.
* Any colour except black.

* Should withstand rainfall, heat/temperature and wind.
* Should be structured in such a way that the trap is escape proof.
Specification of Lures:
1. Lure made of sulphur free rubber/polypropylene/PVC, Impregnated with specific

pheromone blends.
2. Field efficacy should be minimum for 15 days after application.
3. Impregnated lures should be packed singly in individual trilaminated pouches with 30 M1

Aluminum foil.
4. Shelf-life of Lure in original pack should be minimum 6 months at room temperature.
5. Lures should attract insect species only, with 50% insect attractancy by
pheromone/lure/dispenser by using wind tunnel method.
Toxicology
S. Parameters Semiochemicals (pheromones)

No
9(3b) 9(3) 9(4)
1. R R No data required if
Acute oral toxicity- rat
chemical equivalence
2. Acute dermal toxicity- rat R R is established with
9(3) registrant
3. Acute Inhalation – rat R R
4. Primary Skin Irritation - R R
Rabbit
5. Primary Eye Irritation - Rabbit R R
6. Skin Sensitization - Guinea pig R R
7. NR R
Sub- acute oral- rat
8. NR R
Sub-acute dermal
9. NR R
Sub- acute inhalation
10. NR R
Neurotoxicity
11. R R
Teratogenicity
12. NR R
Effect on Reproduction
13. NR R
Carcinogenicity/chronic
toxicity/combined toxicity
study*
14. NR R
Metabolism
15. R R
Mutagenicity (AMES + 2 in-
vitro +1 in-vivo)*
16. R R
Cellular immune response
17. Toxicity to bird (2 species)* R R
18. Toxicity to fresh water fish R R
19. Toxicity to Honey bees NR R
Note: 1. LepidoptereanPheromones that are naturally occurring compound designated by an

unbranched aliphatic chain (between 9 and 18 carbons) ending in an alcohol, aldehyde or
acetate functional group and containing upto 3 double bonds in aliphatic backbones can
be exempted from the sub-acute toxicity carcinogenicity, effect on reproduction and
metabolism if their use rates do not exceed 150 gm/acre/year with Good Agricultural
Practices (GAP) and used in solid matrix dispensers.
2. For formulation products no toxicitydata are required unless it is added with some other
pesticides.
3. * PLEASE REFER TO GUIDANCE DOCUMENT ON TOXICOLOGY FOR

REGISTRATION OF CHEMICAL PESTICIDES IN INDIA.

registration of Semiochemicals / Pheromones on merit case to case basis.
scientifically sound weight of evidence approach and read across/bridging from structurally
and /or biologically related similar pesticides specifically case to case basis on full merits.
3) The replacement alternatives not involving experiments on animals would only be
considered in exceptional cases, in case of alternatives if available with full and sound
justification is provided specifically case to case basis on merits subject to the full
satisfaction of the expert regarding toxicity data provided for the alternatives.
PACKAGING
Chapter V of the Insecticides Rules 1971 in the Insecticides Act, 1968, the rule 16 to 20
of the said chapter deals with the Packaging and Labeling.
Sl. Parameter Semiochemicals

No. (pheromones)
9(3b 9(3) 9(4)

)
1. Labels and Leaflets per IR-1971, all fields (as R R R
applicable) and as amended from time to time
2. Manner of labeling and Leaflet R R R
3. Type of packaging R R R
(Ultra small, small or Big whichever is
applicable)
4. Manner of packaging R R R
5. Specification for primary, Secondary and R R R

Transport packages (whichever is applicable)
6. Details of packaging material and its R R R

compatibility with content
7. Performance of container with content during R* R R
storage stability test(Shelf life Study)
8. Transport worthiness test R* R R
Note:
1. In case of additional packaging endorsement applications, the data at Sl. No. 05, 06, 07& 08, are
not required if similar packaging (material) is being sought by the applicant as has been granted
to earlier 9(3) registrant.
3. All Packaging tests must be carried out with the product of same batch and in its commercial
package preferably in Indian condition.
5. Storage stability data should be generated keeping at least the following parameters in test
protocol such as test temperature, test duration, test packaging material, content of active
ingredient (a.i.) and relevant impurities in the product during and after storage, test humidity,
exposure to light, physical and chemical properties of the product during and after storage etc.
6. The testing protocols must have their basis in the WHO/FAO/ CIPAC/ASTM recommendations
or other validated methodology of GLP/ NABL accredited laboratory having packaging testing
(chemical / mechanical as applicable etc.) in the scope.
7. The Accelerated storage study (ASS) test must be conducted at 54± 2oC (wherever applicable) for
14 days as per FAO/ WHO manual for claiming appropriate shelf life of the product which can be
maximum two years, subject to the condition of providing the ambient storage stability study data
of thirty months or as the case may be within thirty months from the date of application for the
registration.
The abbreviations used in the guidelines are in APPENDIX- I
1. ASS: Accelerated Storage Stability

2. ASTM:American Society for Testing and Materials
3. ATR: Analytical Test Report
4. CFU: Colony Forming Unit
5. CIL: Central Insecticide Laboratory
6. CIPAC: Collaborative International Pesticides Analytical Council
7. CRM: Certified Reference Material
8. CR: Certificate of Registration
9. CSIR: Central Scientific Industry and Research
10. DNA: Designated National Authority
11. ELISA: Enzyme Linked Immunosorbate Assay
12. EPN: Entomopathogenic Nematode
13. FAO: Food and Agriculture Organization
14. FI: Formulation Import
15. FIM: Formulation Indigenous Manufacturing
16. FI-WRT: Formulation Import without registering Technical
17. FIM-WRT: Formulation Indigenous Manufacturing without registering Technical
18. GAP: Good Agriculture Practices
19. GLP: Good Laboratory Practices
20. GV: Granulosis Virus
21. ICAR: Indian Council of Agricultural Research
22. ICMR: Indian Council of Medical Research
23. IR: Insecticides Rules
24. ISI: Indian Standards Institutions
25. LC: Lethal Concentration
26. LD: Lethal Dose

27. LLIN: Long Lasting Impregnated Nets
28. MIC: Minimum Inhibitory Concentration
29. MOH&FW: Ministry of Health and Family Welfare
30. MOU: Memorandum of Understanding
31. MRL: Minimum Residue Limit
32. MUP: Manufacture Use Products
33. NABL: National Accreditation Board of Testing and Calibration Laboratories
34. NBAIR: National Bureau of Agriculture Insect Resources
35. NIBSM: National Institute of Biotic Stress Management
36. NPV: Nuclear Polyhedrosis Virus
37. NR: Not Required
38. PGR: Plant Growth Regulators
39. POB: Polyhedral Occlusion Bodies
40. PVC: Poly Vinyl Chloride
41. R: Required
42. RC: Registration Committee
43. SAU: State Agriculture University
44. SDAS: State Designated Agencies
45. TC/TK: Technical Grade
46. TI: Technical Import
47. TIM: Technical Indigenous Manufacturing
48. VCRC: Vector Control Research Centre
49. WHO: World Health Organization
The terms used in the report are defined in APPENDIX- II
1. Active ingredient: The part of the product that provides the pesticidal action.
2. Applicant: The party (manufacturer, importer or their representative) that makes an

application for registration of a pesticide to the responsible authority.(Please refer to
Form-1 foot note under the Insecticide Rule 1971)
3. Bio-pesticides : Biopesticides is a generic term generally applied to a substance derived

from nature, such as a microorganic or botanical or semiochemical that may be
formulated and applied to control the pest and diseases.
4. Contaminant or impurity in MPCA: Any microorganism or substances it produces that

are present in a product, other than the specified microorganism (or substances it
produces) of the microbial pest control agent (MPCA); an alternate/mutant form of the
MPCA is considered to be a microorganism impurity.
5. Equivalence : The determination of the identical similarity of the purity, impurity and
toxicological profile as well as of the physical and chemical properties, presented by
supposedly similar technical material originating from different manufacturers/place.
6. Formulation: The combination of various ingredients in order to render the product

useful and effective for the purpose claimed in the manner recommended.
7. Good Laboratory Practice (GLP): A quality system concerned with the organizational
process and the conditions under which non-clinical health and environmental safety
studies are planned, performed, monitored, recorded, archived and reported.
8. Herbicide, an agent, usually chemical, for killing or inhibiting the growth of unwanted
plants, such as residential or agricultural weeds and invasive species.
9. Household pesticides are commonly used indoors to control pests such as ants,
cockroaches, houseflies, mosquitoes, fleas, ticks, bedbugs, termites, rodents, mites and
microbes. ... Household pesticides may contain one or a combination of active ingredients
of synthetic (chemical) or natural (plant or microorganism) origin.
10. “Import” means bringing into any place within the territories to which this Act extends
from a place outside those territories;
11. Infective or Infectivity: The ability of a microorganism to invade and persist in a viable
state and to multiply within or on an organism, with or without disease manifestation. The
nature of an infection can vary widely with respect to severity, location and number of
organisms involved
12. “Insecticide”(Pesticide) means - -

i) Any substance specified in the Schedule; or
ii) Such other substances (including fungicides and weedicides) as the Central
Government may, after consultation with the Board, by notification in the
Official Gazette, include in the Schedule from time to time; or
iii) Any preparation containing any one or more of such substances.
13. “Label” means any written, printed or graphic matter on the immediate package and on
every other covering in which the package is placed or packed and includes any written,
printed or graphic matter accompanying the insecticide.
14. “Manufacture” in relation to any insecticide, include :-

i) Any process or part of a process for making, altering, finishing, packing labeling,
breaking up or otherwise treating or adopting any insecticide with a view to its
sale, distribution or use but does not include the packing or breaking up of any
insecticide in the ordinary course of retail business; and
ii) Any process by which a preparation containing an insecticide is formulated.
15. “Package” means a box, bottle, casket, tin, barrel, case, receptacle, sack, bag, wrapper or
other thing in which an insecticide is placed or packed.
16. Pathogenicity: The ability of a microorganism to cause disease and/or inflict damage on
the host. Many pathogens cause disease by a combination of (i) toxicity and invasiveness
or (ii) toxicity and colonizing ability. However, some invasive pathogens cause diseases
that result from an abnormal reaction of the host’s defense system.
17. Pesticide product: The formulated product (pesticide active ingredient(s) and co-
formulants) in the form in which it is packaged and sold.
18. Plant growth regulators (PGRs) are chemicals used to modify plant growth such as
increasing branching, suppressing shoot growth, increasing return bloom, removing excess
fruit, or altering fruit maturity.
19. Plant extract/concentrate: a botanical substance produced from the defined source(s) and
by the described manufacturing processes, and which is the “active substance”. For
botanical active substances, the extract will be in most cases a mixture of components
from the plant and in addition all components that result from the cultivation, harvest,
post- harvest storage and primary processing and manufacturing. It may be difficult to
identify and characterize all individual components. Some of these components may be
considered as components of concern which may be considered in the same way as
“relevant impurities” in chemical pesticide.
20. Post-harvest :In agriculture, postharvest handling is the stage of crop production
immediately following harvest, including cooling, cleaning, sorting and packing etc.
21. Public Health pesticides: Pesticides that are used in the control of pests of public health
significance under public health programs in the country.
22. Registration Dossier: The set of data that is submitted by applicants, in a structured
manner, in support of their application for registration. as per the requirements of the
registration committee.
23. Semiochemicals: Chemicals emitted by a plant or animal that evoke a behavioral or

physiological response in another organism. When the semiochemical affects an
individual of the same species, it is called a ‘pheromone’. When it affects an individual of
a different species, it is called ‘allelochemical’.
24. Technical material: Technical-grade materials and technical concentrates; also known as
technical-grade active ingredient (TGAI).
Technical grade of MPCA: Microbial material used for manufacture of microbial pest control
products. It is the purest preparation of the MPCA resulting from a typical production process,
and contains no additives except for purposes of MPCA growth or replication, or typical
purification and preparation. It may be commercially distributed to manufacturers of microbial
pest control products either in its pure form or augmented with preservatives, stabilizers, and
diluents; or it may be a hypothetical stage in the manufacture of the microbial pest control
product.

Guidelines CIB RC

Uploaded by

Copyright:

Available Formats

Guidelines CIB RC

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Guidelines CIB RC

Uploaded by

Copyright:

Available Formats

t

F.N o. 19 -65 | 2022-CIR-I

1. THE REQUIREMENTS/GUIDELINES FOR REGISTRATION OF INSECTICIDES

1. Form – I duly signed/digitally signed

2. Copy of BOD Resolution/Affidavit/Partnership deed (Notarized)

3. Affidavit for Chemical composition on NJSP (Notarized )

4. Certificate as per category of Industry/ Manufacturing license (Notarized)

5. PAN No. (Notarized)

6. Incorporation Certificate (Notarized) and other documents as per KYC requirement

7. Proof of ownership/lease agreement of manufacturing site purported to be used as manufacturing site

9. Name of registrant of Formulation Import (Reference of Registration Committee Meeting in which

DATA REQUIREMENTS FOR CHEMICAL PESTICIDES

Data Requirements for Chemical Pesticides:

3. Physical and Chemical R R R R R R R NR R NR NR

7. Analytical Test Report R R R R R R R NR R NR NR

10. Shelf-life claim R R R R R R R R R R R

11. Storage Stability Data R R R R R R R NR NR NR NR

12. Establishment of NR NR NR NR R*/ R*/ R*/ R R R R

13. Detailed stepwise R R R R R R R NR R NR NR

14. Information about Raw R R R R R R R NR R NR NR

15. Effluent Treatment R R R R R R R NR R NR NR

16. Legalized letter of R NR R R NR R R R NR R NR

17. Registration Certificate R NR NR R NR R NR R NR R NR

18. In case the supplies are to R NR NR R NR R NR R NR R NR

the invoice would

19. An In-Process sample to R R R R R R R NR R NR NR

20. Methodology for residue R R R R R R R NR NR NR NR

Remarks on chemistry parameters:

Sr. Parameters 9(3b) 9 (3) 9(4)

Remarks on Bioefficacy parameters:

6. In case of Formulation Import without registering technical or Formulation Indigenous

(i) Fate and behavior in treated crop produce

(i) Registration status of MUP in other countries.

c) Data requirements for Registration of formulation –

2) An applicant seeking registration of a formulation without registering technical for

1. Data on bio-effectiveness For label claim of controlled atmospheric

Remarks- Appropriate placement and comments from ICAR\NCIPM\SAU

1. Bioefficacy data generated by ICMR / MOH&FW Institutes based on multi-centric three

A. Bio-efficacy claims In case of Formulation:

a. Bio-efficacy claims to be given on the labels as under:

b. Bio-efficacy claims to be given on the leaflets:

i. Bio-efficacy test on the proposed formulations should be conducted under Indian

E. Data requirements for the registration of new formulations of the approved

Sl. Parameters Technical Formulation Technical/

TI/TIM TI/TIM FIM FIM/FI TIM TI/FIM/FI

2. Acute Dermal- Rat/ R R R R NR NR

9. Repeated dose 90 days R R NR NR NR NR

10. Repeated dose dermal R R NR/R NR/R NR NR

13. Repeated dose R R NR NR/R NR NR

16. Developmental R R NR NR/R NR NR

20. Two generation NR R NR NR NR NR

22. Pharmacokinetics and R R R R NR NR

26. Acute Avian toxicity * R R R R NR NR

28. Avian Reproduction R R R R NR NR

29. Acute toxicity to Fresh R R R R NR NR

31. Acute Toxicity- R R R R NR NR

32. Medical data R R R R NR NR

33. Human toxicity NR NR NR R NR NR

34. Health records of NR R NR R NR NR

12. Establishment of NR NR NR NR R/ R/ R*/ R R R R