Progress 07/01/23 to 06/30/24

Outputs
Target Audience: Nothing Reported Changes/Problems:During this reporting period, we encountered several delays and challenges that affected the progress of the project. One of the most significant issues involved hardware and software problems with the Opentrons OT-2 liquid handler, which slowed down our high-throughput screening process. These technical difficulties resulted in a reduced number of experiments being conducted, with only about 300 completed thus far. Additionally, a lab shutdown at the Melotto Lab at UC Davis, one of our key partners, due to renovations, further delayed our ability to send samples for analysis. This shutdown impacted our timeline for collecting and processing critical data. As a result, the rate of expenditure has been slower than anticipated, since key phases of the project, such as optimization, have not yet begun. Despite these delays, no changes have been made to the approved Data Management Plan. Our redesign of the database structure, which ensures compatibility with machine learning models, remains within the original project scope. However, the research schedule has experienced deviations due to the technical challenges and the UC Davis lab shutdown. The screening phase, which was expected to be further along, is still in progress, and this has postponed the optimization phase for improving the mechanical and antimicrobial properties of the candidate materials. Nonetheless, we remain confident that we will be able to recover the lost time in future periods with the help of our upgraded equipment and improved software. So far, there have been no unexpected scientific outcomes. While no standout antimicrobial candidates have been identified yet, this is consistent with the early stage of the screening process, and we anticipate identifying more promising results as the experiments continue. Additionally, there have been no changes to the approved protocols for the use or care of animals, human subjects, or biohazards during this reporting period. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, AEMS Corp will take several actions to overcome the delays encountered in the current phase and continue advancing the project goals. First, we are addressing the challenges in our high-throughput screening process by upgrading our hardware and enhancing our software. We are in the process of upgrading our OT-2 liquid handler to a Flex, which will eliminate failed experiments caused by incorrect liquid level detection, thus increasing reliability. Additionally, the software we are developing integrates design of experiments into the scripting, which will allow us to design and execute experiments more efficiently. These improvements will enable us to make up for the lost time and significantly accelerate the pace of data collection. Once sufficient data is collected, we plan to train machine learning algorithms to identify molecular trends that can improve the mechanical properties of the antimicrobial coatings. This insight will allow us to refine formulations by doping the most promising compound mixtures to enhance their durability and efficacy. The machine learning-driven approach will provide an efficient path to optimizing the coatings, making them more robust for practical applications in food processing environments. Since we have already redesigned our database structure to be fully compatible with machine learning models, we anticipate no issues in implementing this next phase. We are also addressing the challenges posed by external factors. The Melotto Lab at UC Davis, which is a key partner for sample processing, was closed for several months due to renovations, delaying our ability to send samples for validation. However, to mitigate this, we have relied on data from a related project involving organic sanitizers to identify a preliminary formulation that shows efficacy against E. coli O157. This allows us to move forward with collecting evidence of our coating's effectiveness, albeit using a suboptimal organic acid formulation. By limiting the number of tests performed during this period, we will still be able to collect the data necessary to demonstrate that the coating works as intended. Overall, the combination of software enhancements, hardware upgrades, and strategic adjustments in our experimental approach will allow us to address the issues from the current period while maintaining progress towards achieving our goals. We expect to complete the necessary screenings and begin optimization in the next reporting period, positioning us to further validate the antimicrobial efficacy of our coatings and continue developing a commercially viable solution for the food processing industry.

Impacts
What was accomplished under these goals? This project addresses the urgent need to reduce cross-contamination of deadly pathogens, particularly E. coli O157, in food processing facilities. These pathogens contribute to widespread foodborne illnesses, resulting in public health crises and costly product recalls. To address this issue, we are developing novel, food-grade antimicrobial coatings made from Generally Recognized As Safe (GRAS) organic acids. These coatings will offer continuous protection during food processing, thereby improving food safety and reducing contamination risks. Our work directly benefits food processing facilities by providing them with a sustainable solution to contamination issues, potentially lowering their operational costs associated with recalls and sanitation. Additionally, food safety regulators will gain new tools to enhance their safety enforcement protocols, while consumers will enjoy greater confidence in the safety of their food products. The development of coatings that can be applied during active food processing operations ensures an ongoing shield against contamination, making our solution particularly effective in high-risk environments. During this reporting period, we made progress toward two major goals. First, we initiated high-throughput screening of GRAS-based antimicrobial materials. We purchased and implemented the Opentrons OT-2 liquid handler and developed custom software to run the screenings. However, technical challenges related to hardware and software delayed our progress, limiting us to about 300 experiments so far. The screening is still in its preliminary stages, and no standout candidates for antimicrobial activity have been identified. Despite these challenges, we have established a strong foundation for future screenings and anticipate accelerating the pace of this work in the coming months. Our second goal, the optimization of the mechanical and antimicrobial properties of candidate materials, is on hold until we complete more comprehensive screenings. While this delay has postponed optimization efforts, it will not affect the overall project schedule. Once we identify viable candidates, we will begin refining their mechanical properties to ensure the coatings are both durable and effective in preventing cross-contamination. Key accomplishments during this reporting period include the successful establishment of a high-throughput screening system and the completion of 300 initial tests on GRAS ingredient combinations. While we have not yet realized any major breakthroughs in antimicrobial activity, the project remains on track. The insights gained from overcoming early challenges have refined our approach, positioning us to make significant progress in the next phase. As we continue, we anticipate identifying promising candidate materials and moving forward with optimization and validation in simulated industrial conditions.

Publications