Background: Some plants produce toxic chemicals or antimicrobial agents to protect themselves; these could be synthesised to create medicines. Much current research is dedicated to attempting to find cures or medicinal properties through these plants.
Objective/Purpose: To test if certain plants have antimicrobial functions or properties.
Materials: Balance, weight boat, lab scoops, LB broth base, Media bottles 250 mL, Sterilizer/autoclave, Water bath 37*C shaking, Sterile LB agar, Laminar flow hood and disinfectant, Glasses safety plastic, Bunsen burner and gas lighter, Inoculating loop, Ni/Cr wire, Petri dishes 60x15mm sterile, E. coli JM109 (stock plate), Plant specimen, Mortar and pestle, Pipet 10 mL and pump, Plastic funnels short stemmed, Filter paper disks 5 mm diameter, Beakers 100 mL, Syringe 10 mL and filter 0.2um, Reaction tubes and rack 1.7mL, Methanol absolute, Pipet 1 mL and pump, Dry block heater/heat block, Forceps fine-tipped, Ampicillin, Glass spreader, Incubator oven 37*C
Procedure: Using a mortar and pestle, grind up two grams of plant tissue with 10 mL of deionized water. Let it sit for 3 minutes. Filter the sample through a filter paper/funnel. Label the sample. Repeat the first steps, using methanol instead of water. Attach the prefilter to the syringe and rinse with water. Take these to the hood, and use the pipette to transfer the sample to the syringe. Filter the solution through the syringe into a microfuge tube, close the tube, and label. Label agar plates with a cross on the bottom, label quadrants as well.
Results:
24 Hours: A small ring has appeared around the pieces of paper, most are not very noticeable. No signs of contamination and the bacterial lawn is thick and off white. The negative control has no ring but the positive control has a significant clearing.
48 Hours: The rings on the methanol test have widened slightly, but the water test papers have not changed. Controls have not changed.
72 Hours: Nothing has changed.
Analysis/Conclusion: The positive control disk (the ampicilin, located in quadrant 4 in pictures 1 and 3 above) was the only disk to have any major clearance around it, as it was most likely the only compound in the petri dish to have any antimicrobial properties. However, the bacterial lawn was thin and difficult to see; it did not grow well over the agar gel and the clearings were difficult to discern. The negative control (the water disk, located in quadrant 4 in pictures 2 and 4) had no clearing, and our weak bacterial lawn grew right up to the filter paper. Obviously, water would not have antimicrobial properties because it had no chemical or biological compounds that could deter, harm, or disable any known bacteria. Our plates were most likely not contaminated, and we saw slight clearings around our plants extracts that had been created with methanol but none that had been create with water. This could mean that the plants were extracted better with this compound, or that the disks had some extra methanol in them. Our plants would not have held up to a stronger bacteria, but perhaps could have prevented the growth of a weaker one. Our results go to show that our plant (wild rose) had very weak antimicrobial properties.
Objective/Purpose: To test if certain plants have antimicrobial functions or properties.
Materials: Balance, weight boat, lab scoops, LB broth base, Media bottles 250 mL, Sterilizer/autoclave, Water bath 37*C shaking, Sterile LB agar, Laminar flow hood and disinfectant, Glasses safety plastic, Bunsen burner and gas lighter, Inoculating loop, Ni/Cr wire, Petri dishes 60x15mm sterile, E. coli JM109 (stock plate), Plant specimen, Mortar and pestle, Pipet 10 mL and pump, Plastic funnels short stemmed, Filter paper disks 5 mm diameter, Beakers 100 mL, Syringe 10 mL and filter 0.2um, Reaction tubes and rack 1.7mL, Methanol absolute, Pipet 1 mL and pump, Dry block heater/heat block, Forceps fine-tipped, Ampicillin, Glass spreader, Incubator oven 37*C
Procedure: Using a mortar and pestle, grind up two grams of plant tissue with 10 mL of deionized water. Let it sit for 3 minutes. Filter the sample through a filter paper/funnel. Label the sample. Repeat the first steps, using methanol instead of water. Attach the prefilter to the syringe and rinse with water. Take these to the hood, and use the pipette to transfer the sample to the syringe. Filter the solution through the syringe into a microfuge tube, close the tube, and label. Label agar plates with a cross on the bottom, label quadrants as well.
Results:
24 Hours: A small ring has appeared around the pieces of paper, most are not very noticeable. No signs of contamination and the bacterial lawn is thick and off white. The negative control has no ring but the positive control has a significant clearing.
48 Hours: The rings on the methanol test have widened slightly, but the water test papers have not changed. Controls have not changed.
72 Hours: Nothing has changed.
Analysis/Conclusion: The positive control disk (the ampicilin, located in quadrant 4 in pictures 1 and 3 above) was the only disk to have any major clearance around it, as it was most likely the only compound in the petri dish to have any antimicrobial properties. However, the bacterial lawn was thin and difficult to see; it did not grow well over the agar gel and the clearings were difficult to discern. The negative control (the water disk, located in quadrant 4 in pictures 2 and 4) had no clearing, and our weak bacterial lawn grew right up to the filter paper. Obviously, water would not have antimicrobial properties because it had no chemical or biological compounds that could deter, harm, or disable any known bacteria. Our plates were most likely not contaminated, and we saw slight clearings around our plants extracts that had been created with methanol but none that had been create with water. This could mean that the plants were extracted better with this compound, or that the disks had some extra methanol in them. Our plants would not have held up to a stronger bacteria, but perhaps could have prevented the growth of a weaker one. Our results go to show that our plant (wild rose) had very weak antimicrobial properties.