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Team: Moscato for Mosquitoes

Team Lead: E. Fox

Team Members: G. Nigonawoe | K. Anderson | Dr. V. Dougnon | C. Brew,

Sustainable Mosquito Control – Safe Biopesticide Production

We plan to utilize local agricultural waste products to produce viable biopesticides in mosquito infested regions via rudimentary fermentation technology.

Define the problem:

Current methods of mosquito control can be costly, potentially dangerous, and inaccessible to regions most in need. Biopesticides, such as Bacillus thuringiensis subsp. Israelensis (Bt/Bti), have been used to effectively control mosquito populations. However, commercial Bt/Bti production is considered expensive. Overall, cost-effective use of biopesticides would lessen the nuisance factor imposed by invading mosquito populations.

Describe your big idea:

The use of the bacteria Bacillus thuringiensis subsp. Israelensis, as a biopesticide is a proven and safe concept. This bacterium produces proteins which are particularly toxic to the larvae of Aedes and Anopheles mosquitoes (known contributors to vector-borne human diseases). Ease of mosquito control is essential in reducing the number of humans affected by disease. Developing sustainable production of a Bt/Bti biopesticide would allow us to globally combat mosquito larvae and reduce disease occurrence. While the technology to ferment Bt/Bti is commercially available, it is generally expensive and inaccessible.

Overall, effective application of rudimentary fermentation technology to economically/viably produce Bt/Bti biopesticide which would lessen disease occurrence and the nuisance factor imposed by invading mosquito populations.

Describe/illustrate proposed solution:

  • Examine cassava/yam waste as culture media for Bt/Bti

  • Construct rudimentary fermentation units

    • Feed with locally sourced cassava/yam waste (rinsed/mashed/sterilized)
    • Augment with locally available sugar
  • Introduce pure Bt/Bti culture to fermentation units

    • Monitor and manually agitate fermentation units for consistent growth and sporulation
    • Utilize microscopy and polymerase chain reaction (PCR) analyses to confirm purity
    • Produce cost-effective/environmentally friendly biopesticide which reduces mosquito populations
    • Final product could be distributed to mosquito infested waters to reduce mosquito larvae populations.

Simplified Overview:

Process

The core foundation of your research or solution:

7,500+ scientific manuscripts explore the use of Bt/Bti to control mosquitoes. Previous research/commercial application of Bt/Bti has proven its successful use in controlling mosquitoes. Bt/Bti is not harmful to humans, vertebrates, or ecosystems, has few impacts on non-target organisms, and is naturally biodegradable. Due to expensive culture media, current commercially available fermentation technologies cannot cost-effectively produce Bt/Bti. Less than 1,000 manuscripts have explored alternative growth media for Bt/Bti. Exploration of less expensive culture media demonstrates that potatoes can be used as a cheap alternative for culture media in Bt/Bti production. Traditional fermentation units/bioreactors are expensive and require the attention of a trained technician. Without sacrificing gross product efficacy, we propose the use of commonly available materials to construct rudimentary fermentation units. We hypothesize that large water containers such as 208-liter barrels will suffice. Using black containers in warm climates should allow sunlight to bring the internal temperature up to the necessary 27-37-degree C temperature for bacterial sporulation (acting biopesticide component). Utilization of widely available cassava or yam waste has not yet been applied in rudimentary fermentation technology to produce Bt/Bti.

Describe your end-user and/or paying customer:

The primary end-users will be landowners and/or those affected by mosquito infestation. Payment sought from local municipalities for regional product distribution.
This project will be tested and implemented in rural/developing regions. Local students/community members will be trained in microscopy/molecular skills. These are transferable academic/research/industrial skills. In the long-term, this project has scalability potential for regional economic development. The produced product may be sold to regional communities/municipalities.

Describe Technology Readiness/Research Literature Levels.

Please see details above. Optimal technology exists, as does ample opportunity to improve the overall process. We're at a TRL of 2-3.

Sampled related manuscripts:

DOI: 10.1016/j.toxicon.2006.11.022, DOI: 10.1002/jobm.200510585, DOI: 10.1023/A:1014937311839

Describe three critical hypotheses:

H1. Locally sourced agricultural waste, such as cassava/yam, will prove to be a viable growth medium for Bt/Bti.

H2. Rudimentary fermentation units constructed from widely available materials (i.e. 208-liter barrels) adequately function to grow bacteria.

H3. Cassava/yam waste, as measured in H1, when implemented in rudimentary fermentation units constructed in H2, will adequately grow Bt/Bti and produce a biopesticide product.

Hypotheses Testing/Technology/Phases:

H1 examined via laboratory experiments. Previously unexplored root vegetable products will be mashed, emulsified with a sugar, and autoclaved before plating/filling petri dishes/tubes. Bt/Bti bacteria introduced to the plates/tubes and growth compared to control samples (standard nutrient agar/broth). Growth viability measured, reported, and results will dictate application in H3.

H2 examined via laboratory environment. Previously unexplored application of rudimentary fermentation units will utilize at least three 208-liter black plastic barrels will be procured and sterilized. Standard nutrient broth will be introduced to 2 of the barrels, and 1 will remain a control unit. A cosmopolitan bacterium will be introduced into barrels 1 and 2. Barrels placed in sunlight, temperature/conditions monitored, liquid samples will be retrieved and analyzed via microscopy techniques for bacterial growth. Monitored conditions and growth results dictate application of rudimentary fermentation in H3.

H3 utilizes results from H1 and H2. Bt/Bti will be grown, monitored for purity (pathogen free), and sporulation via microscopy. Parameters for optimal sporulation will be developed before material is harvested and utilized as a biopesticide. Biopesticide can be delivered to local municipalities and/or landowners for application.
Future studies will need to evaluate the use of the produced biopesticide in field application. Additional metrics related to student training and the potential use of this technology to promote local economic growth needs to be explored.

Describe how would use funding to progress your hypotheses:

Budget Breakdown

Project Gantt Chart

What you’ve done to date:

Progress To Date

Why your idea is an unconventional or creative approach to the problem.

As indicated, significant research has been conducted on Bt/Bti . Limited manuscripts have explored the use of agricultural waste as growth support media for Bt/Bti. Our project will build on current knowledge, exploring the use of cassava/yam as a growth medium. We’ve hypothesized previously unexplored rudimentary fermentation technology which will produce a cost effective Bt/Bti biopesticide.

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