What is the significance of using Salmonella Typhimurium as a model organism? And how applicable is this to other types of bacteria as far as the effectiveness of the compounds as a therapy method?
Salmonella Typhimurium is relatively cheap and safe and it has a short incubation period. It is also a good model in rodents because it models a more serious infection that the more dangerous bacteria Salmonella Typhi causes in humans.
Results are somewhat applicable to other bacteria, but most especially gram negative and rod shaped bacteria. This is because the tested compounds would use the same mechanism for killing one type of bacteria as another.
It is much more complicated to determine the effectiveness of compounds as a therapy method. With Salmonella Typhimurium in particular, inside of an organism, the bacteria invades macrophages and survives within them by matching their pH with that of the macrophage. The tests performed in these initial experiments simply test Salmonella in M9 media, but do not show whether the compounds tested would be effective in a macrophage, much less an entire organism.
More testing would have to be done to determine this. First, it would be important to perform more max dose experiments to obtain a more valid statistical result to affirm this. Since it was first only determined whether the compound worked, this result could be due to many factors such as experimental error or any unreliable mechanism of the compound.
This was done, but various errors occurred in the process of doing so, which led us to decide to switch to solid media in testing Alpha Pinene. However, only concentrations lower than the 10% concentration used in the max dose experiment could be used because the human body cannot absorb any compound at more than 10% concentration. With this, it would be unlikely for the Alpha Pinene to be more effective at lower doses, but this is not completely impossible. Furthermore, since it was only effective about 50% of the time in the max dose experiments, it would be better to continue testing such as this in the more effective 1 ,8 Cineole.
DMSO models the conditions of the body well and is known to not be antibacterial or antimicrobial. It could then be assumed to not kill the Salmonella and used as a negative control. However, it was not a good negative control for the dilution series with the 1 ,8 Cineole because the 1, 8 Cineole was not soluble in it, so no data could be collected, leading us to use the Mueller-Hinton agar plates. In the agar plates, it was placed into the well alone, so there were no conflicts with solubility therefore allowing it to still be used.
Could you elaborate more on why the results of the Dilution series were excluded, how was Alpha Pinene incorrectly diluted and do you think that this component had any significant effect on your final results? Also, do you think that the problems with solubility that you mentioned will effect future research?
What is the significance of using Salmonella Typhimurium as a model organism? And how applicable is this to other types of bacteria as far as the effectiveness of the compounds as a therapy method?
Salmonella Typhimurium is relatively cheap and safe and it has a short incubation period. It is also a good model in rodents because it models a more serious infection that the more dangerous bacteria Salmonella Typhi causes in humans.
Results are somewhat applicable to other bacteria, but most especially gram negative and rod shaped bacteria. This is because the tested compounds would use the same mechanism for killing one type of bacteria as another.
It is much more complicated to determine the effectiveness of compounds as a therapy method. With Salmonella Typhimurium in particular, inside of an organism, the bacteria invades macrophages and survives within them by matching their pH with that of the macrophage. The tests performed in these initial experiments simply test Salmonella in M9 media, but do not show whether the compounds tested would be effective in a macrophage, much less an entire organism.
Why was the effectiveness of Alpha Pinene 50% but not 100%. What limited its effectiveness to 50%.
More testing would have to be done to determine this. First, it would be important to perform more max dose experiments to obtain a more valid statistical result to affirm this. Since it was first only determined whether the compound worked, this result could be due to many factors such as experimental error or any unreliable mechanism of the compound.
Could Alpha Pinene be tested at differing concentrations to increase it’s affect
This was done, but various errors occurred in the process of doing so, which led us to decide to switch to solid media in testing Alpha Pinene. However, only concentrations lower than the 10% concentration used in the max dose experiment could be used because the human body cannot absorb any compound at more than 10% concentration. With this, it would be unlikely for the Alpha Pinene to be more effective at lower doses, but this is not completely impossible. Furthermore, since it was only effective about 50% of the time in the max dose experiments, it would be better to continue testing such as this in the more effective 1 ,8 Cineole.
Why did you choose DMSO as your negative control?
DMSO models the conditions of the body well and is known to not be antibacterial or antimicrobial. It could then be assumed to not kill the Salmonella and used as a negative control. However, it was not a good negative control for the dilution series with the 1 ,8 Cineole because the 1, 8 Cineole was not soluble in it, so no data could be collected, leading us to use the Mueller-Hinton agar plates. In the agar plates, it was placed into the well alone, so there were no conflicts with solubility therefore allowing it to still be used.
Could you elaborate more on why the results of the Dilution series were excluded, how was Alpha Pinene incorrectly diluted and do you think that this component had any significant effect on your final results? Also, do you think that the problems with solubility that you mentioned will effect future research?