DOSE RESPONSE CURVES OF ESSENTIAL OIL COMPOUNDS IN POSTHARVEST SPOILAGE PREVENTION OF FRESH PRODUCE

WP 1: Method development, validation, and test planningThe objective for WP1 is to define, develop and validate the experimental set up and techniques. This WP will involve development of the test chambers, methods to control and measure the EOC concentrations in the gas phase, methods to monitor and measure the growth and spread of B. cinerea infections on fruits as well as various quality attributes of the fruits.Thus, the tasks included in this WP are:Definition of the EOCs and selection of their target gas-phase concentrations to be tested. This portion of the plan will be informed by the EOC screening and characterization experiments that are being carried out separately by our bioactive formulation development team within StixFresh. The EOCs will be chosen from a subset that are shown to be effective against B. cinerea and we are most likely to commercialize as determined at the time of selection.Detailed definition of the use case to be simulated and definition of the initial conditions of the produce. For example, in an effort to standardize the starting conditions, produce may first be sanitized in a bleach solution and then dipped in solutions containing known concentrations of the target spoilage organism. In Phase I, we plan to test the effectiveness of EOCs against Botrytis cinereaDefinition and development of the experimental techniques that will be required to support the testing phase in WP2.Development and validation of techniques for "volatilizing" the target EOCs quantitatively so as to approximate the target gas-phase concentrations in the test environments. This effort will be informed by the thermodynamic and physicochemical properties of the EOCs (for example vapor pressure as a function of temperature) when possible. Whenever possible, estalished or previously published techniques such as quantitative injection with a syringe combined with heat evaporation will be employed.Development and/or adaptation of techniques measuring the head-space EOC concentrations in the gas phase with GC-MSMicrobiological methods to characterize the in-vitro growth, and extent of infection and spoilage of produce in-vivoMethods for characterizing the relevant quality attributes of the produce such as %Brix or total soluble solids and firmness.Design and build of any test chambers, fixtures etcPreliminary experiments as needed to validate the methods chosen and verify the workflows.Development, review, and approval of the detailed test plan.WP 2: Detailed testingThe objective for WP2 is to obtain the gas-phase dose response curves (DRC) of candidate EOCs under the select use case conditions both in-vitro and on fruits by executing the test plan developed in WP1. This objective will involve testing the effect of EOCs across a range of gas phase concentrations at 5 concentration levels on the suppression of growth and spread of B. cinerea inoculated in growth media (in-vitro), as well as on fruits (in-vivo). Potential target produces of interest include bananas, strawberries, blueberries, cherries, grapes, kiwi fruit, mangos, peaches, oranges, and avocados. The test systems chosen include a selection from this set, to be determined at that time based on the most relevant fruit(s) we are likely to target with our first product to market.WP 3: Results analysis, reporting, and recommendationsThe objective for WP3 is to analyze, synthesize and capture the learnings from the project. This work package will focus on detailed analysis and interpretations of the results obtained in WP 2. Results will culminate in a narrative for recommendations for further R&D as well as design guidelines for the follow-on product development activities in Phase II.