BME.02 – High Throughput Anaerobic Intestinal Microbiota Screening Tool

• Abhi Krishan
• Ruby Martinez
• Sharon Roberts
• Michelle Sanchez
• Miles Woodcock Girard

Microbiologists studying the human intestinal microbiota encounter the challenge of individually testing numerous bacterial strains, which is crucial for advancing research into patient-tailored, microbiota-based therapies. Current solutions, such as the BioLector bioreactor, automate the culturing process to a degree, but are limited by short culturing times and low throughput. To address this, we have developed a high-throughput, small-scale anaerobic chemostat system capable of culturing and performing multiple simultaneous pharmacological screening trials on intestinal microbiota strains. This system consists of a fluidic lid that is designed to be compatible with standard 24 well culture plates, with the ability to run 6 experiments in parallel, while guaranteeing precise experimentation conditions. It integrates a series of microcontrolled valves and pumps to meticulously facilitate the transport of nutrients, metabolites, and prospective therapeutic agents, controlling inflow and outflow necessary for screening experiments. Critical requirements were identified, focusing on maintaining precise flow rates within a specified range of 3.0 to 4.0 mL/sec, with a deviation tolerance of ±10% for both inflow and outflow of medium/nutrient supply channels. Additionally, requirements were outlined to uphold intra- and inter-cyclic sterility, emphasizing the importance of autoclavable system components to mitigate contamination risks. These specifications are crucial for guaranteeing the system’s dependability and efficiency in facilitating microbiota-based therapies. Verification testing was conducted to assess flow rate consistency via a terminal-based command interface. The device’s pump was tested over time intervals ranging from 1.5 to 10 seconds and incrementing by 0.5 seconds, with 25 trials for each of the 17 intervals, and the volume of water pumped during each interval was measured. A line of best fit was then obtained for these data, the slope of which represented the average flow rate over all trials. Results indicated an average flow rate of 3.56 mL/s, with an average standard deviation of 0.004 mL/s. With a deviation of 0.112%, the device demonstrated highly consistent flow rates well with the acceptable range outlined by the pass criterion. Additionally, the results exhibited an R^2 value of 0.999, signifying the near-linearity of the verification data and emphasizing the system’s consistent delivery of flow rates. With this high-throughput bioreactor device, we hope to further enable microbiologists in their research towards advancing personalized treatments for patients. By providing consistent and reliable flow rates within the specified range to supply the bioreactor, our system holds the potential to significantly streamline the pharmacological testing of multiple bacterial strains, ultimately paving the way for more effective and personalized management of gastrointestinal conditions.