The seminar on Problem Formulation for the Risk Assessment of Biopesticides stemmed from a previous CRP-sponsored event on Innovating Microbial Pesticide Testing that identified the need for an overarching guidance document to determine when in vivo tests are necessary. Problem Formulation, a common practice in pesticide risk assessment, was highlighted as a useful approach for addressing uncertainties in data requirements for biopesticides. The seminar featured presentations from various perspectives, including industry, regulatory bodies, and academia. Topics included the history and principles of Problem Formulation, industry perspectives on Problem Formulation and how it is applied internally for microbial pesticides, regulatory approaches, and specific case studies. The seminar provided an overview of the challenges, considerations, and potential solutions in harmonising Problem Formulation for biopesticide risk assessment. It emphasised the need for collaboration and discussion to develop Problem Formulation guidance for biopesticides.

1. Part 2:
Case Study: Peptides-based Plant
Protection Product (harpin proteins*)
Rosa Criollo (FMC Corporation)
on behalf of the CLE Biopesticides Expert Group
Emmanuelle Bonneris (Bayer)
on behalf of the IBMA Natural Substances Professional Group
*Disclaimer: public information from regulatory bodies as US EPA has been used as reference information

2. Introduction
1. EC Reg 283/2013 and 284/2013-Part B were developed for microorganisms; however, any other biopesticides
technology fall by default in Part A of EC regulations 283 & 284.
2. The EU industry associations (CLE & IBMA) have developed specific tools to address data requirements suitable for
novel technologies with the aim to support the availability of such new technologies to farmers.
a) IBMA has developed a “Data decision tree for identifying potential risks for natural substances when used in
plant protection”, to support the regulatory data (guideline) development and then data evaluation by
highlighting the relevant potential risk areas for natural substances. IBMA presented the Natural Substances
Decision Tree including 3 case studies in DG SANTE’s Biopesticides Working Group (6 June 2023).
b) CLE drafted a “Contribution document on the assessment of innovative biochemicals”, with the objective to
identify what data and risk assessments requirements are suitable for those technologies. CLE presented the
document in DG SANTE’s Biopesticides Working Group (Nov-22 and Nov-23). It compiles lessons learnt from
the data requirements data package from EC Reg 283/2013- and 284/2013, and from other regulatory
bodies to address the human health and environmental safety of any novel technology.
3. In this exercise, one of the CLE case studies (e.g., “Peptides based technology") was chosen for presentation and
compared to the IBMA Decision Tree to explore its applicability.
4. From the IBMA SURVEY of 26 member companies completed in March 2023, 9 peptides both identical to nature (5)
or modified (4) were communicated as ready for submission in the EU between 2023 and 2028.
5. End of 2023, CLE has launched an anonymous SURVEY of 6 questions to map out the potential number and type of
"biochemicals" (e.g., proteins, enzymes, RNA, etc.) that could enter in the EU market in the coming years. The
objective is to communicate aggregated figures to the EU COM and Member States to encourage development of
a fit for purpose regulatory framework for such novel technologies.

3. Case study: Harpin protein
• In a nutshell the reference case:
• “Harpin protein” is a microbially produced peptide. Commercially produced, is identical to
the protein that occurs in nature and produced under high quality-controlled conditions.
• Registered active ingredient: Three harpin protein are registered in USA as PPP Ag
use since the year 2000 [PC code 006477, 006506, 005200 and PDHP 25279]
• Registered in Europe under national law (e.g. France as fertilizer)
• Mode of Action (MoA) and biological properties: It acts through the ability to reach and bind
with its target site/organism with high specificity, and the MoA is non-toxic. It initiates a
complex set of metabolic responses in the treated plant, causing systemic gene expression
and eliciting a plant’s natural growth and defense system → the plant is an elevated
resistance to bacterial and fungal infection, suppression of nematode egg production as well
as increased growth and higher yield. This protective response makes the plant resistant to a
wide range of fungal, bacterial, and viral diseases during growth and post-harvest.
• Formulation: 1% Harpinαβprotein formulated as a wettable granule
• Application:
- Foliar spray; seed treatment; irrigation/chemigation; pre-plant dip; greenhouse soil
drench.
- Use rates are generally 0.14-3.69 grams of active ingredient per acre at approximately
14-day intervals.

4. Data Decision Tree Approach for
natural substances - basis for a
framework and concept
Data decision tree for identifying potential risks for natural substances (NS) when
used in plant protection, published in Taylor and Francis Biocontrol Science and
Technology, 6 June 2023:
https://www.tandfonline.com/doi/full/10.1080/09583157.2023.2210268
Use as Plant Protection Product => Possible Exposure
❑humans (workers, operators, bystanders, residents, consumers)
❑crops and harvested products
❑organisms inside and outside the treated areas
Risk dependent on:
❑substance identity and character
❑background levels
NS should have minimal to no adverse effects to nontarget organisms, humans, ecosystems
and the environment.
NS: In general, complete degradation and/or decline to existing background level

5. Data Decision Tree Approach for
natural substances - basis for
assessment of relevant risk areas
• Branched Data Decision tree i.e., 'branches from 1 to 5", developed to focus on relevant
risk areas to address, with relevant data requirements for natural substances
• Depend on 'group-definition' adapted from guidance documents submitted by FAO/WHO
(2017)* and OECD (2017)**
• Efficacy (EU requirements): see EPPO, 2017
• Every decision taken in Data Decision Tree must be supported by good quality data
(published literature or well-documented studies).
(*https://www.fao.org/3/i8091en/i8091en.pdf; **https://doi.org/10.1787/31f295f3-en)

6. e.g., Human Toxicology
2.1 Group 1 Harpin protein Notes
2.1 Does the NS belong to
Group 1*?
No
Proceed to step 2.2
Yes
to step 2.1.1
Yes
*Group 1 categorization: ‘NS that
are, with current knowledge, known
to have no established effects on
humans, animals and the
environment with recognized
specifications.
2.1.1 Data/published literature to address acute toxicity
2.1.1 Is data/published
literature available to
properly address acute
toxicity?
No or only partly
Address acute toxicity.
Proceed to step 2.1.2
Yes
No further studies needed.
to step 2.1.2
Yes
Human Tox data package already
available (6 pack)
2.1.2 Food/feed use
2.1.2 Does the NS have a
food/feed use or any other
registered use that is
based on a risk assessment
(e.g., biocide use)?
No
Conduct risk assessment.
To Branch 3
Yes
Proceed to step 2.1.3
No
Already registered in US.
Registered as fertilizer in France
2.1.3 Exposure similar/lower than via food consumption
2.1.3 Is the exposure following
PPP use similar to, or
lower than, food
consumption or is it non-
existent?
No, exposure following PPP use
is significantly higher than via
food consumption.
Conduct risk assessment.
To Branch 3
Yes
No further studies nor risk
assessment needed.
to Branch 3 Data Decision
Tree
No
Food diet consumption unknown ->
need a qualitative risk assessment
(lit. search)
Legend: NS: Natural Substances ; "Branch 3": Residues

7. Recommendations:
- Tailored data requirements/guidance document(s) specific for novel technologies in the
OECD work program (e.g. Guidance Document on peptides).
- The continuation of ongoing initiatives to adapt the risk assessment framework for existing
technologies such as botanicals to ensure it is fit for purpose (RATION, EFSA Grant).
Conclusions on the tool developed
by IBMA for the CLE peptide case
1. The IBMA data decision tree is a valuable tool for identifying potential areas with the use of risk-
based flow chart. The guidance on how to address these endpoints or how to provide the
required data need further development.
2. The CLE approach focuses on identifying what studies (data package) would be the most
suitable scientific data to address the data requirements in the EU for biochemical pesticides in
general, and per individual novel technologies (peptides, RNAi, etc).
3. These two tools are complementary and provide a good starting point for authorities to further
developed guidance documents. Both CLE and IBMA initiatives are under the "need to know"
principle.

8. Thank you very much
for your attention
*Disclaimer: public information from regulatory bodies as US EPA has been used as reference information