Objectives: The objective is to set up the experimental conditions to isolate and to test the biological properties (antimicrobial and enzymatic properties) individually and in synergy of a protein isolated from the hemolymph (blood) of honey bees subjected to individual and combined pathogens / stressors.
Abstract: Domesticated bees have been subject to much research into a broad range of complex wellbeing issues for the last 20 years, but this has failed to lead to the development of concrete solutions for beekeepers. Up to today, the research and methodologies deployed against new challenges (such as climate and flora changes, increased use of pesticides, prevalence of viruses, Nosema spp, bacteria and parasites like Varroa destructor), have focused on evaluating and attempting to separately prevent and fight each factor. Complex colony population dynamics and poor understanding of the immune response of bees have so far made it impossible to identify acceptable tolerance thresholds vs. combinations of stress agents. Since several years our laboratory is working on this theme, with aim to address the above limitations by establishing robust, effective, sensitive and hyphenated analytical technologies for profiling & deciphering bee immune-proteomes with regards to the host-pathogens-symbionts-environment interactions. We have recently isolated a protein from the hemolymph of the honey bee that have similarities with a family of protease inhibitors. As this protein may represent an interesting marker to follow during infection, we would like to quantify its activities in the context of different infectious conditions.

Methods: The isolation of protein of interest from hemolymph of honey bees will be performed by HPLC and its identity confirmed by mass spectrometry. The biological properties will be evaluated with antimicrobial assays (antibacterial, antifungal) and enzymatique activity measurement.
Relevant publications of the team:
Pisani C, Voisin S, Arafah K, Durand P, Perrard MH, Guichaoua MR, Bulet P, Prat O. (2016) Ex vivo assessment of testicular toxicity induced by carbendazim and iprodione, alone or in a mixture. ALTEX. 2016;33(4):393-413.
Herren JK, Paredes JC, Schüpfer F, Arafah K, Bulet P, Lemaitre B. (2014) Insect endosymbiont proliferation is limited by lipid availability. Elife. 2014 Jul 15;3:e02964.
Bulet P & Arafah K (2014) Proteomics in Methods in Immunology, from theory to practice Collège des enseignants d’immunologie (Assim) Coordinators: Marie-Christine Béné, Christian Drouet, Sylvain Fisson et Estelle Seillès

Requested domains of expertise (few keywords): Biochemistry, mass spectrometry, high performance liquide chromatography, microbiology.

Laboratory: UGA-IAB Inserm 1209, CNRS 5309 Director: Pierre Hainaut
Team: Equipe 8 Head of team: Patrice MARCHE
Name and status of scientist in charge of the project: Philippe BULET, DR CNRS, HDR yes

Address and person of contact:
Plateforme BioPark d’Archamps, Bâtiment le Forum, Archamps Technopole, 74160 Archamps
Phone: 0450432521, e-mail: [email protected], [email protected]

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