Why do you think you saw such a large effect on the growth in Figure 2 at a 10 µM concentration but not at the max dose in water concentration in Figure 1?
This is unclear. We were not able to replicate the effect from the figure 2 dilution series, with either the max dose experiment or the agar plate. The result from figure 2 was anomalous. However, we can be pretty sure that thymoquinone would not be a good antibiotic. Even if the figure 2 result is taken at face value, the lower doses (ie. ones which could actually be used in a clinical setting) are not effective.
You mentioned problems with the solvents on the plate that didn’t show results — what kind of problems are these? Why do you think these solvents didn’t work well and what were the major differences between this plate and your successful plate?
We were not having solvency problems on the plates. We actually started using agar plates because we wanted to get away from the solubility issues we were having in the other experiments. The issue was basically that thymoquinone was not dissolving (or dissolving very slowly) in the two solvents we used, water and DMSO. It is still unclear if thymoquinone is truly soluble in either substance.
It’s interesting that low doses of Thymoquionone seemed to actually INCREASE the growth of S. Typhimurium compared to the negative control. I am not familiar with the measuring method used; is it possible that the Thymoquionone itself could be absorbing some light and skewing the results? Could there be a control of Thymoquionone with NO bacteria to make sure of that.
We did not do a control to see the absorbance of a thymoquinone solution without bacteria. I would be very surprised if thymoquinone by itself had a significant effect on the absorbance. Thymoquinone is a very small chemical, and even at higher doses it would probably not block enough light to effect the absorbance. However, if I were to redo the experiment, I would probably check the absorbance on a straight thymoquinone solution just to be sure.
The most important question we had at the end of this experiment was whether longifolene would be effective against salmonella in solvents such as water and DMSO, not just in an agar plate.
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Why do you think you saw such a large effect on the growth in Figure 2 at a 10 µM concentration but not at the max dose in water concentration in Figure 1?
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This is unclear. We were not able to replicate the effect from the figure 2 dilution series, with either the max dose experiment or the agar plate. The result from figure 2 was anomalous. However, we can be pretty sure that thymoquinone would not be a good antibiotic. Even if the figure 2 result is taken at face value, the lower doses (ie. ones which could actually be used in a clinical setting) are not effective.
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You mentioned problems with the solvents on the plate that didn’t show results — what kind of problems are these? Why do you think these solvents didn’t work well and what were the major differences between this plate and your successful plate?
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We were not having solvency problems on the plates. We actually started using agar plates because we wanted to get away from the solubility issues we were having in the other experiments. The issue was basically that thymoquinone was not dissolving (or dissolving very slowly) in the two solvents we used, water and DMSO. It is still unclear if thymoquinone is truly soluble in either substance.
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It’s interesting that low doses of Thymoquionone seemed to actually INCREASE the growth of S. Typhimurium compared to the negative control. I am not familiar with the measuring method used; is it possible that the Thymoquionone itself could be absorbing some light and skewing the results? Could there be a control of Thymoquionone with NO bacteria to make sure of that.
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We did not do a control to see the absorbance of a thymoquinone solution without bacteria. I would be very surprised if thymoquinone by itself had a significant effect on the absorbance. Thymoquinone is a very small chemical, and even at higher doses it would probably not block enough light to effect the absorbance. However, if I were to redo the experiment, I would probably check the absorbance on a straight thymoquinone solution just to be sure.
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What kind of future research do you think would be most effective in advancing your research?
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The most important question we had at the end of this experiment was whether longifolene would be effective against salmonella in solvents such as water and DMSO, not just in an agar plate.
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