Periodic Reporting for period 1 - IPM-4-Citrus (IPM-4-Citrus, Citrus desease Integrated Pest Management: from Research to Market) Reporting period: 2017-04-01 to 2019-03-31 Summary of the context and overall objectives of the project IPM-4-CITRUS aims to develop two new bio-pesticides active against citrus pests and scale them up from lab to market. It is considering the potential of 2 alternative endemic strains (Bt kurstaki BLB1 and LIP) to produce a biopesticide (as alternative to chemical treatments) and to protect citrus from 2 targeted pests (Phyllocnistic citrella, Prays citri). This project strengthen collaborations between academic and non-academic partners based in 3 European Member States (France, Germany and Italy), 2 Associated Countries (Turkey and Tunisia) and 1 Third Country (Lebanon),The project’s research and innovation activities are based on a multidisciplinary approach, which aims at understanding and sensitizing stakeholders about the health risk factors related to citrus pests and developing an alternative Integrated Pest Management (IPM) approach based on biological control. In conjunction with validation through field tests, the project will pave the way for future commercial exploitation of these new biopesticide products by drawing up a feasibility study for future spin-off activities and/or new production lines in partner SMEs. This project will then give a competitive advantage to the consortium partners as well as Europe and enhance the chances of job creation.Staff secondments and inter-sector and international mobilities between complementary partners will represent a unique opportunity to optimize bioproduction processes and obtain high added-value bioproducts, while building up the partners’ skills and reinforcing the training of early-stage researchers through knowledge sharing and networking. The project will also adopt a concrete RRI (favouring public engagement, informal education). Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far In WP2 (Bioproduction), our consortium conduct an exhaustive investigation of literature and experimentation based on rational cultivation strategy (WP2) using competencies from LISBP, CBS, FS-USJ, TWB, MEDIS. From M0 to M24, the following actions have been conducted: • Identify reference and compare all investigated culture media and associated performances,• Development and harmonisation of Material and Methods, • Screening of bioproduction condition at lab scale. Identification of the optimal substrate (wheat bran vs granulometry), comparison of 3 strains under standardized culture conditions in flask• Report bio-kinetics, titer and yield for delta-endotoxin production in flask,• Transposition to bioreactor (3 and 5L) • Investigation of alternative metrology to monitor cell growth and endo-toxin production, • Validation of a semi-synthetic medium for comparison of bioperformances in bioreactor, • Model the sporulation mechanism with Bt kurstaki which is oriented to parasporal crystal (final product).In WP3 (Bioactivity and Formulation), the optimised process protocol will allow the production of pilot lots whose formulation formula, approved at lab and field levels, will be validated using JKI skills (formulation), and BBU and CTA facilities (biopesticide efficiency test). WP3 was just initiated at M18, preliminary results are: • Simple and low-cost downstream process to extract/separate endotoxin from low cost raw material in suspension in broth, • Formulation in dry-way after cultivation with semi-synthetic medium, • Investigation of the activity of the formulated product (with standard formulation conditions) on both target pests, in labs and in the fieldIn WP4, the analysis of the product’s value chain will sustain the project’s pertinence from lab to field and will be finalised by an economic maturation and technology transfer approach (from research to market). The core of this maturation activity (WP4) is driven by joint collaboration between academic and private and leaded by BIPCA. WP4 run throughout the project. At present time, a market, technological and regulatory assessment has been conducted. Four potential innovations have already been identified that ensure the IP value of the project and could increase the chances of investor interest. Several results were shared within consortium and disseminated through public engagement and informal education through the different outreach activities. Various events open to a larger audience have been conducted: • TP1 Training (20th April 2018, CBS, Sfax – M12): Biocatalyst improvement & bioreactor cultivation, from basic concept up to intensified bioproduction• TP2 Training : Industrial microbiology, fermentation and scale-up - Cell cultivation in bioreactors (M14 and M17) @ CBS• TP2 Training: Industrial microbiology, fermentation and scale-up - Cell cultivation in bioreactors (M23) @ USJ (repetition)• RT1 Round table: Scientific roundtable including external scientific experts: state-of-the-art of Bt culture; comparing & sharing experiences (8th March 2019, CBS, Sfax – M22)• RT3 Roundtable (20th April 2018, CBS, Sfax – M12)• WS1: Workshop with external expert (7th March 2019, Leban)• Visit of Biopesticide Start-up (Taanayel, Lebanon)• PE1: participation in ESOF 2018 (Toulouse, 11th July 2018) • Contribution to other events : o Rencontre5plus5-IPM-4-Citrus-, 7April2017 (Tunis, Tunisie) managed by IPTo Turkish Consumer Dissemination-Byians-seminar, April2018 (Ankara, Turkey)o European Researcher's Night-IPT-Sep2018, (IPT, Tunisia)o Foire des sciences, Bierut-LEB, March2018 (USJ, Lebon)o 17th International Days of Biotechnology IDB 208, December 20-23,2018 (Sousse, Tunisia)o 2nd Mediterranean Congress on Biotechnology (MCB2), 16-20 March 2019 (CBS, Hammamet) Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) Beyond the state-of-the-art and first results, we intend to demonstrate that an alternative and low-cost biopesticide can be produced considering Bioproduction, Formulation-Bioactivity and Tranfer steps. IPM-4-Citrus consortium leads to identify 4 dimensions of innovation: 1. Robust fermentation process with low cost raw material (associated with simple bioprocess without pH regulation, and cheap formulation after a simple decantation) by using 2 alternative strains (Bt kurstaki BLB1 and LIP respectively Tunisian and Lebanon endemic strains), 2. Alternative configuration of instrumentation for real time process monitoring (objective : process monitoring and control) 3. Standards & norms for Bt production process for the MENA & Sub-Sahara African countries (control and regulation issue specification/methodology guided regulation cost reduction to access to the market) 4. Innovative application strategy for controlling lifeminer insects through epiderm (formulation)Through the development and transfer of innovations from the lab to the market, IPM-4-CITRUS aims to facilitate knowledge sharing, help young scientists to acquire new skills and develop their career owing to the recommendations of the MSCA-RISE-2016 call. It will contribute to expand private sector opportunities for SMEs but also for young graduates by giving them a chance to explore opportunities in entrepreneurship, master business planning, market analysis, business modelling, marketing, market entry. The Responsible Research and Innovation aspects of the project are demonstrated through different organized round tables & workshops, specialised events attended and websites. These communication and outreach activities tools have a wide impact since they target a large audience composed of scientific stakeholders but more importantly citizens at large who are interested in the topic of biopesticides.