Performing Department
(N/A)
Non Technical Summary
Humans use fresh water for drinking, irrigation, producing energy, and industrial activity. Globally, over 70% of freshwater is used for agriculture to produce crops, which provide 30-40% of the world's gross food output. With the increasing growth of the global human population, freshwater usage is projected to grow to meet the needs of growing demand for crops. It is inevitable to seek alternative sources of water, such as recycled water and agricultural return flows, to guarantee food and water security. However, poorly monitored recycled water and return flows can transmit waterborne diseases to consumers. If the presence of pathogen(s) can be identified, specialized corrective countermeasures could be executed to increase the quality of return flows and recycled water for irrigation, thereby quenching the increasing need for freshwater usage. To mitigate such a large-scale problem, technologies are needed to address the following question: How can we assure the quality of return flow and reclaimed water?In this proposal, we will address this question by pairing C60 probes that target 16S rRNA sequence for identification with microchips that multiplex the tests. Biospecimens collected from farms and water treatment plants will also be tested. We aim to (i) develop C60 probes for waterborne pathogens, (ii) develop and validate microchips for multiplexing waterborne pathogens, and (iii) validate novel efficient methods for sterilization. This proposed technology will be able to profile multiple pathogens simultaneously in 15 minutes from end-to-end cost-effectively.
Animal Health Component
50%
Research Effort Categories
Basic
10%
Applied
50%
Developmental
40%
Goals / Objectives
Humans use fresh water for drinking, irrigation, producing energy, and industrial activity. Globally, over 70% of freshwater is used for agriculture to produce crops, which provide 30-40% of the world's gross food output. With the increasing growth of the global human population, freshwater usage is projected to grow to meet the needs of growing demand for crops. Therefore, it is inevitable to seek alternative sources of water, such as recycled water and agricultural return flows, to guarantee food and water security. However, poorly managed recycled water and return flows can transmit waterborne diseases to consumers, currently, we do not have a cheap, efficient, and easy-to-use solution package to identify if there is ongoing contamination, what is the source of contamination if there is one, what is the best solution for the confirmed contamination, and how can we measure the efficacy of the treatment? Therefore, our overarching goal is to address the fundamental yet significant question: "How can we assure pathogen-free recycled water and return flows?" To accomplish this overarching goal, we have a list of specific objectives to accomplish this goal. The objectives include1. Use bioinformatic analyses to determine the signature sequences for a list of ten waterborne bacteria: including Shigella, Yersinia enterocolitica, Campylobacter, Pseudomonas syringae, Pseudomonas cepacian, Leptospira, Vibrio cholerae, Ralstonia solanacearum, Xanthomonas campestris, Xanthomonas axonopodis.2. Design detector, mismatch, and reporter sequences based on bioinformatic analyses.3. Synthesis C60 probe and validate the synthesized probes with chemical characterizations.4. Determine the efficacy of each C60 probe with corresponding bacteria.5. Design and synthesize microfluidic that concentrates bacteria by syringe.6. Design and synthesize microfluidics for multiplexing bacteria with verified C60 probes.7. Test and validate microfluidics' efficacy in identifying multiple bacteria simultaneously.8. Investigate the effect of PDI with PdC14 against waterborne pathogens.9. Determine the optimized PDI with PdC14 in reclaimed water.
Project Methods
Our efforts are:Training graduate students in identifying and mitigating waterborne pathogens.Training undergraduate students by offering a new elective topic: water-transmitted diseases.Changing the knowledge of genetic copies of a positive detection does not necessarily translate to viable bacteria contamination for a particular case.The PI and Co-PIs should follow each one's milestones and deliverables. The milestones are:Expand cell culture by Month 6.Design detector, mismatch, and reporter sequences by Month 12.Synthesize and validate C60 probes using designed sequences by Month18.Determine the efficacy of all ten C60 probes with corresponding bacteria by Month 24.Evaluate the stabilities of all C60 probes in different liquid conditions by Month 24.Fabricate and test microfluidics with different designs by Month 12.Test the selected microfluidics in the laboratory by Month 18.Test the selected microfluidics with wastewater by Month 24.Test the combination of C60 probes with microfluidics in the lab with wastewater by Month 36.Test the Efficacy of PDI in the lab by Month 24.Test the efficacy of PDI with wastewater effluents by Month 36.