Biomolluscicide microbial strains for the protection of plants from invasive snail and slug species.

Invasive species have caused more than $1 trillion in damage, with more than $420 billion annually and are considered one of the top contributors to the earth's significant and rapid decline in biodiversity, right behind land-use alteration. Invasive species are getting worse and we do not do enough to prevent and manage invasive species. Gastropods (e.g., slugs and snails) have been estimated to be responsible for at least $3 billion of this figure, with three gastropods listed among the worst 100 invasive species in the world: the golden apple snail ($1B annually in rice in Phillipines), the giant African snail, and the rosy wolf snail. Additionally, invasive slug species have been linked with up to $100 million in damage to Oregon's grass seed business and observers have seen entire crops of mint wiped out by slugs.Countries around the world are losing billions to crucial crops, such as rice, citrus, flowers and herbs, among at least 500 other plants.Solutions for preventing crop loss from invasive slugs and snails are currently limited to toxic chemicals and new, safer options are therefore required for plant and crop protection from invasive slug and snail species.In order to combat such a major agricultural pest, Invasive Species Corporation team plans to develop their existing base microbe-based technology into a commercially viable product using precision fermentation and formulation platform technologies. ISC team is energized by USDA team's acknowledgement of the problem for food and agriculture and awarding this grant to proceed with the much-needed biological solution. This Phase I proposal is highly responsive to USDA research priorities, including SBIR Topic 8.2 "Plant Production and Protection (Biology)", and is particularly relevant to the agency's interest in "innovative tools and management practices" and "alternative chemicals or biology- based strategies" for the protection of crop species. Ensuring that invasive snail and slug speciesare unable to damage US agriculture is clearly within the agency's remit and advances one of its core missions.The goal of this USDA Phase I SBIR is to evaluate the efficacy of a microbial molluscicide against invasive slug and snail species for the protection of high-value crop plants such as citrus trees, home gardeners and landscapers. Invasive Species Corporation (ISC) is developing a formulation of the Pseudomonas fluorescens strain CL145A (Pf CL145A), which is known to have high molluscicidal activity against invasive quagga and zebra mussels owing to a key membrane protein calledfitD.While previous studies have shown activity against these targets, a more potent and cost effective product/formulation is needed. ISC team plans to optimize current technologyto reach a cost that will allow commercialization of a product for slug and snail control. During Phase I, fermentation will be optimized to ensure high, bioactive titers of Pf CL145A, and a prototype bait formulation will be developed that enables long-lasting protection of crop species as well as high efficacy against the most damaging slug and snail species.Through this grant, ISC is addressing the development ofnovel agents capable of addressing the currently unmet need for safe, effective, and scalable solutions for invasive snail and slug infestations.Several chemical products exist viz. metadehyde andferric phosphate (Gusto IRON), however they either have human and animal toxicity associated with their use and/or are not environmentally safe. Both these products require hazardous warning labels by EPA. Currently, none of these solutions is manufactured using regenerative methods (such as bacterial fermentation to be used by ISC's product). Phosphate run-off has been associated with eutrophication in the waterways. ISC team believes thata product based on the molluscicidal activity of PfCL145A and its active ingredient FitD protein, would represent a safer, more efficient, effective protective agent, preventing agricultural losses and protecting yields while reducing environmental and health risks to the surrounding community and end-consumer. For home gardeners and landscapers, customers will have a more efficient product, a true biodegradable biological that is easier to use and based on what we know about the PfCL145A, without such label restrictions as iron phosphate.ThisPhase I program is designed toestablish the feasibility of using PfCL145A and/or its active agent FitD protein as a cost-effective and efficient molluscicide for the eradication of invasive slug and snail species infestations. Phase I plans to maximize the dose of FitD protein generated from PfCL145A manufactured using state-of-the-art fermentation technologies, as well as to verify the effect of the microbe and its active protein against gastropod species. These efforts will validate ISC's strategy and set up pre-commercialization efforts in Phase II. In Phase I, the studies will include·fermentation and formulation optimization to maximize the production of fitD protein and/or fermentation productivity of active biomass through cost-effective processing to maintain fitD activity during processing. Formulation efforts will focus on preserving the activity of the biomass containing active molecule while delivering the product to targets in a bait and slow release granules. The final prototype product will go through in-silico/in-vitro study to show its non-toxicity to non-target species. These studies will establish the feasibility of ISC's approach to snail and slug invasive species mitigation and lead to a Phase II program designed to complete safety and efficacy testing for future registration with EPA and subsequent commercialization. Successful efforts in Phase I will enablePhase II activities which would include 1)field trialswith private CROs, such as Syntech and Agmetrics, with university researchers and on farm grower demonstrations, 2)mammalian and ecotoxicology studiesas required by the EPA for use in agriculture, and 3) manufacturing scale-up and quality assurance testing.