Final Report Summary - INNOVA (Innovative bio-based pesticides to minimize chemical residue risk on food) CONTEXT, GENERAL AIM AND SPECIFIC OBJECTIVES OF THE PROJECT INNOVACrop protection measures, which include chemical pesticides, have significantly contributed to productivity and quality of produce worldwide. Integrated Pest Management (IPM) programs have become the major strategies in modern crop protection since many years in Europe. However they are mostly based on empirical approaches addressing the exploitation of short term solutions to reduce pesticide residues on food. The minimum residue level is fixed in Europe for each type of pesticide and food or feed (Regulation (EC) No. 396/2005; Regulation (EC) No 178/2006; Regulation (EC) No 299/2008 and following amendments) and there should be no risk of exceeding these limits if the good agricultural practices are fulfilled and pre-harvest interval is respected. However, several chemical pesticides have a long pre-harvest interval and re-entry period and long residual effect, therefore protecting crops close to harvest is often challenging. In particular, several crops are harvested for long periods of time (i.e. tomatoes, peppers, strawberries, vegetable crops, etc.) with frequent picking (often daily) and most of chemical pesticides cannot be applied. Therefore, there is a strong need for innovative products from natural origin, safe for humans and the environment, renewable and with a fast biodegradation on the crops in order to minimize residues on food. These products are especially important in those crops where the harvest time is long and plant protection is needed. One of the main bottlenecks in developing these products is represented by the poor overlapping and integration between public research institutes and private companies, the first mainly focusing on the production of fundamental scientific knowledge, the latter specialized in the production, formulation and selling of products. Therefore the main aim of this proposal was to reach a better integration between public research and private sector by a close collaboration specifically oriented to industrial and social needs. In this context the specific objectives of the project INNOVA (logo) were:1. To identify innovative biopesticides to help growers to produce pesticide residues-free products focusing of the most critical crops and growing stages in term of risk of pesticide residues on final food and to select the potential biopesticides according their intrinsic technological qualities.2. To identify the suitable formulations of the innovative biopesticides, in term of shelf life and efficacy against pests and to scale-up the production.3. To deepen the research on some specific aspects requested for the registration of the new bioproducts. 4. To identify the best application strategies in IPM and the suitable market strategies.WORK CARRIED OUT IN THE PROJECT INNOVAThe first two years of the project have been devoted to identify innovative biopesticides 1) to help growers to produce pesticide residues-free products, focusing on the most critical crops and growing stages, in term of risk of pesticide residues on final food and 2) to select the potential biopesticides according their intrinsic technological qualities. The crops with the higher risk of chemical residues (number of residues and their concentration, exceeding/not exceeding the MRL) and the target pests/diseases have been identified based on public reports (EFSA Journal) and database (FAO stat, EU pesticide database) and with the input of the expertise present in the companies. A large number of potential biopesticide candidates, both of the Partners of the project and identified by an ‘active search’ in scientific literature and by contacting technology transfer offices in the Academic sector. The new innovative biopesticide candidates have been listed and classified in term of potential efficacy against the selected pests/diseases based on the existing knowledge on mechanism of action, genetics, behavior in the environment. A total of 47 candidates have been evaluated in this first step. The candidates have been then classified in term of way of production, efficiency and cost of production and formulation. In two cases, being them new technologies to be used in combination with the bioproducts, in term of ability to improve their efficacy. A total of 19 candidates have been then selected to be further developed, considering the intrinsic characteristics of the active ingredient and the integration with chemical in strategies to reduce the risk of chemical residues on food. These candidates were: microbial strains and/or their metabolites, botanical extracts, compounds of natural origin and attractant/activators.Preliminary small-scale trials were out both in greenhouse and open field in order to confirm efficacy of the new innovative biopesticides against the selected target pests/pathogens. Ten active ingredients (Fig 1 and Fig 2) were selected for the following development. For each of the ten candidate biopesticides, the most important aspects to be addressed with the formulation (persistence/survival, UV protection, desiccation, temperature degradation, rainfastness, etc.), according to the nature of the biopesticide and the use on the crop, were then identified. In particular, the most important aspects for the microbial agents when applied on the phyllosphere were increasing survival, and decreasing the damages due to UV, desiccation and unsuitable temperature, while increasing survival and multiplication was the most important aspect when applied in soil. Increase of rainfastness and decrease volatilization were the most import aspect for the botanical extracts. To increase and speed up conidia germination the addition of activators was identified as crucial point. For the selected natural molecules, the work focused on co-formulants that can increase the uptake by the pathogens’ cells. A specific carrier to increase the persistency and efficacy in soil of a selected microbial fungicide was designed. Nutritional compounds to increase survival and colonization of pruning wounds and leaves by microbial biocontrol agents were also selected. The possibility to develop a slow-release encapsulation for volatile bio-based botanical extract was explored. Possible lures to increase attractiveness of a candidate insecticide were assessed. The various prototype formulations for each of the ten potential new active substances were compared in term of increase efficacy of the compound against the selected pests and pathogens, in lab trials. A database of suitable products and related formulations in term of industrial production was completed and six candidates were selected for the pilot production (Fig 3). The quality of each batch was assessed in order to check consistency of production. The cost of production of the various prototype formulated products was assessed and compared, in order to select the best production process and formulation.The guidelines for registration of pesticides (Regulation 1107/2009) were followed to verify the feasibility of registration for the six selected candidates. In particular the expected metabolic profile of the selected microbial active ingredients was assessed based on the genome sequencing of the strains and the chemical composition of the plant extracts in the technical grade was analyzed. For each of the six selected products an extensive literature search was carried out, especially in term of possible risk for humans and environment. The mechanism of action of the selected biopesticides was investigated in lab trials. Some small-scale tests (Fig 4) to check eco-toxicology of biopesticides (possible side effects against: microbial populations in soil and phyllosphere with metagenomics approaches, some microbial biocontrol agents with dual culture tests, aquatic organism (Daphnia magna), predators (Aphidius rhopalosiphi, Typhlodromus pyri) were carried out. The fate in the environment was tested for a two microbial biopesticides (colony forming units and quantitative PCR) and the residues in the environment was tested for a natural rare molecule (chemical analysis). Possible side effect on vinification of a microbial biopesticide and a natural rare molecule were also assessed. Five candidates were selected for the small scale trials to define dosage and timing of application and the integration with chemicals. Based on the small scale trials the five candidates were tested with a suitable experimental design following EPPO standards and protocols on strawberry, cucurbits, basil, grapevine, apple, bean and wheat against powdery mildew, downy mildew, grey mold, trunk diseases, Rosellinia root rot and Fusarium head blight, mites, aphids, white flies, trips, leaf miners, Drosophila suzukii (Fig 5) and as desiccant/herbicide. All trials were carried out at least twice, in at least two locations or two seasons/environmental conditions (Fig 6). The results in term of efficacy and reduction of residues in the food were compared with conventional standard pest management strategies (Fig 7). FINAL RESULTS AND THEIR POTENTIAL IMPACT AND USEFour biopesticides (a microbial biofungicide, one plant extracts and two natural molecules) were developed and the registration process as plant protection products (two biofungicides, one bioinsecticide and one bioherbicide) is already started. The registration of the first commercial plant protection products is expected from 2017. It should be completed by the end of 2021 for all the four candidates. Their integration into strategies will allow a significant reduction of residues in food on the most problematic crops. All products are also expected to be allowed for the use in organic agriculture, thus contributing to the competitiveness of this sector.The immediate benefit we achieved is a better coordination of the activities in a strongly fragmented sector like the development of biopesticides, especially between academia and industry. The INNOVA project is also increasing, broadening and enriching the expertise of the researchers involved in the program in all the research areas of developing bio-based plant protection products in a scientific multidisciplinary context. Several results of the project have been already published on peer review journals and presented at scientific conferences. The results related to the four final products have been also disseminated at technical conferences and open-days and in technical journals targeting a public of agronomists, field advisors and growers. The project was presents to students (high school and university) as a successful example of integration of research between Academia and Industry. The planned four-year cooperation also laid down the foundation for a long-term collaboration, to create a virtual platform for the development of new bio-based plant protection products.In Europe the future approach in the agricultural production should be probably radically changed. The bio-economy is a key element also in agriculture for sustainable and green growth in Europe. Concerns on chemical pesticides are continuously increasing. Advancements and innovation in bio-based pesticides research will allow Europe to improve the use of safe, low impact and renewable biological plant protection products besides widening a promising market. The increased replacement of chemical with biopesticides holds a great potential: it can help reducing the use of chemical in agriculture while improving the economic and environmental sustainability of pesticide industries. The development of innovative bio-based pesticides will ensure sustainable resource use and alleviating stress on the environment. The new bio-based plant protection products developed in INNOVA project will be valuable tools to achieve a sustainable use of pesticides in agriculture.The results of this project not are only bringing in-depth scientific understanding of the production of bio-based plant protection products, but also introducing new advanced bio-based active ingredients for possible industrial implementation and production. The main results of the project, namely the four innovative biopesticides, will be important for agriculture and environment, representing safe tools against some of the most relevant pests and pathogens (as white flies, trips, grey mold, powdery and downy mildews, etc.) on important crops as pome fruits, grapevine, soft fruits, Solanaceae, cucurbits, lettuce, etc. and to control weeds.Website: http://www.project-innova.euProject coordinator: ilaria.pertot@fmach.itFigures to be inserted and related caption:LogoFig. 1 Biocontrol agent developed in the project INNOVAFig. 2 Biocontrol agent developed in the project INNOVAFig. 3 Formulated biocontrol agent (wettable granules) developed in the project INNOVAFig. 4 Small scale trials on strawberryFig. 5 Lab trials with a bioinsecticide against Drosophila suzukii Fig. 6 Efficacy trial on tomatoFig. 7 Efficacy against powdery mildew of strawberry of experimental products (1-7 experimental products; 8 sulphur; 9 untreated control)
