Tiny Earth: Essential Microbes as Antibiotics for Model Pathogens
Session Number
Project ID: BIO 11
Advisor(s)
Dr. Tanya Crum, Illinois Mathematics and Science Academy
Discipline
Business
Start Date
20-4-2022 10:05 AM
End Date
20-4-2022 10:20 AM
Abstract
Antibiotic resistance is growing globally, making it necessary for new antibiotics to be discovered. Major pharmaceutical companies have stopped antibiotic discovery, claiming a lack of profitability, so academic institutions are crowd-sourcing this work. We started our research by collecting soil from environments stressful for bacterial survival. We performed a serial dilution of the soil in water to dislodge bacteria from other organic matter, plating the dilutions to obtain soil isolates. Unique bacterial colonies were identified and patched onto master patch plates. Using a modified Kirby-Bauer technique, bacteria were transferred from patch plates onto plates spread with each of the 10 ESKAPE pathogen relatives. If the patched bacteria produced antibiotics, we would see clear zones around each path, indicating that the bacteria can inhibit ESKAPE pathogen growth. The bacteria producing a large zone of inhibition were identified by 16S rRNA sequencing. A region of the 16S rRNA gene was amplified by PCR and sent for Sanger sequencing. We use BLAST to identify each bacteria to at least the Genus level. We also used the gram-staining technique to differentiate and identify the bacteria. Our next steps include possible Eukaryotic toxicity testing and organic extraction and testing of the antibiotic.
Tiny Earth: Essential Microbes as Antibiotics for Model Pathogens
Antibiotic resistance is growing globally, making it necessary for new antibiotics to be discovered. Major pharmaceutical companies have stopped antibiotic discovery, claiming a lack of profitability, so academic institutions are crowd-sourcing this work. We started our research by collecting soil from environments stressful for bacterial survival. We performed a serial dilution of the soil in water to dislodge bacteria from other organic matter, plating the dilutions to obtain soil isolates. Unique bacterial colonies were identified and patched onto master patch plates. Using a modified Kirby-Bauer technique, bacteria were transferred from patch plates onto plates spread with each of the 10 ESKAPE pathogen relatives. If the patched bacteria produced antibiotics, we would see clear zones around each path, indicating that the bacteria can inhibit ESKAPE pathogen growth. The bacteria producing a large zone of inhibition were identified by 16S rRNA sequencing. A region of the 16S rRNA gene was amplified by PCR and sent for Sanger sequencing. We use BLAST to identify each bacteria to at least the Genus level. We also used the gram-staining technique to differentiate and identify the bacteria. Our next steps include possible Eukaryotic toxicity testing and organic extraction and testing of the antibiotic.