Controlo dos efeitos da Paenibacillus larvae em colmeias: efeito combinado de enzimas de fagos com probióticos

The overarching aim of this proposal is to leverage microbial technology to integrate the beneficial effects of probiotics and endolysins for the prevention and sustainable control of American Foulbrood (AFB) in beehives. To achieve this goal, a series of complementary objectives have been outlined: Objective 1 (O1): Conduct an in silico analysis of P. larvae phage genomes to identify endolysins (PLY) with potential antibacterial activity against the causative agent of AFB. O2: Express, characterize, and select the 3 most effective endolysin(s) PLY(s) capable of reducing the load of P. larvae in vitro under various hive conditions. O3: Engineer L. plantarum strains (up to 3) capable of producing the identified PLYs, exogenously (Lpl_PLY) for application in beehives. O4: Obtain a transformant Lpl_PLY widely reducing P.larvae in vitro. O5: Evaluate the dynamics of Lpl_PLY together with Lpa and Ak (Lpl_PLY/PB) in healthy hives following spray administration to confirm its ability to reach larvae and actively combat P. larvae infection. Determine the average concentration of Lpl, Lpa, Ak and PLY reaching larvar in hives. O6: Validate the efficacy of Lpl_PLY/PB, PLY and PBb(Lpl+ Lpa+ Ak) in preventing and/or controlling P. larvae infection, in lab-reared larvae. O7: Demonstrate that the formulated Lpl_PLY/PB positively impacts the immune system and longevity of larval bees in the event of P. larvae infection. O8: Based on O5+O6, extrapolate on the efficacy of the formulation in infected hives, to request that the competent authorities test the product in naturally contaminated hives. Addressing Important Challenges: This proposed research directly tackles the significant challenge of AFB, a devastating disease affecting honeybee colonies worldwide. By integrating probiotics and endolysins, it offers a novel and promising approach for AFB prevention and control, addressing the urgent need for sustainable solutions in beekeeping. Ambitious Objectives Beyond the State of the Art: The objectives outlined in this proposal are ambitious and innovative, incorporating cutting-edge techniques such as in silico analysis, genetic manipulation of microbial strains, and the synergistic use of probiotics and endolysins. This interdisciplinary approach represents a significant advancement beyond current anti- P.larvae strategies, offering novel concepts and approaches to tackle this persistent threat to bee health. By pursuing these objectives, this project aims to revolutionize AFB control in beekeeping, paving the way for more sustainable and environmentally friendly practices while safeguarding the health and vitality of honeybee colonies.