Yeast Busters: Stopping Fungus in its Tracks with Antifungal Medicines Project HELP

[santha123]

Hi Madeline,

The procedure gave an example for a concentration greater than 1% (e.g. 25%) to dilute it to make it a 1% solution and then to follow steps 9b and 9c. But are the rest of my understanding of the procedure correct?

The Oil that I am using does not specify the active antifungal ingredients with percent, but it does say that it's a "fungus fighter."

These are the ingredients listed:
Soybean Oil, Tea Tree Oil, Safflower Oil, Vitamin E, Carrot Extract, Aloe Vera Extract, Rose Hips Oil, Peanut Oil, Sweet Almng Oil, Methylparaben (And) Fragrance, D And C Red #17. D And C Yellow 311

Methylparaben is a fungicide so I think this is the main antifungal ingredient?

and here is the link for the product (TTO):
https://www.walmart.com/ip/Hollywood-Be ... z/10313104

Thanks,
Santha

[MadelineB]

Hello Santha,

Thank you for the link to the Walmart information for your bottle of Tea Tree oil.

When I checked the link that you gave to me, I see they also have "Humco 100% Pure Australian Tea Tree Oil."

I suggest that you use a product which tells you the concentration of the Tea Tree oil. Then, you will need to do a literature search to find out the concentrations that researchers have used, so you know which concentrations to test as the high and which to test as the low concentration. The judges who review your project will want to know that information. Also, remember that you will be putting in a lot of work to measure the outcome, so it would be nice to know what the concentration is that yields the result!

Let us know if you have more questions!

[santha123]

Hi Madeline,

I found a really good scholar page :https://www.google.com/patents/US8333981.
It states how useful tea tree oil is and gives examples of the amount used, but not the dilution. (.06 and .10)

Since the tea tree oil is 100%, I was wondering if I could add 12 tsp to 1/8 of the oil? (25% = 3 tsp)
Then I would do the steps for the low and high concentration.

Just making sure, as far as we have discussed, my understanding of the project procedure is correct?

Thank you so much again for helping all this time. Now I can start my project with a great understanding and no confusion!!!


Thanks,
Santha

[MadelineB]

Hello Santha,
It sounds like you now have a bottle of 100% Tea Tree oil. That will make things easier for you!

However, you will still need to look at the scientific literature to find the starting dilutions for your high dose.

Try googling using a search something like "pub med antifungal effect of tea tree oil."

This should show you open access scientific papers in Pub Med Central so you can read the actual journal papers. You want to find the concentrations of Tea Tree oil reported as "minimum fungicidal concentration."

This is information that you should include in your research report to justify your choice of concentrations to test.

Your patience and persistence will pay off, since the judges review your written report with as much interest as your results!

Let us know if you have more questions.

[santha123]

Hi Madeline,

I found a website (http://www.ncbi.nlm.nih.gov/pubmed/15140856) which clearly states the amount used (1%), so doesn't that mean that I need to dilute it to a 1% concentration( 1/8 tsp oil + 12tsp water)??



Thanks,
Santha

[santha123]

Hi Madeline,

So as far as we have discussed, my understanding of the procedure is good right??? I'm just making sure because I don't want any complications when conducting my experiment.

Thanks,
Santha

[MadelineB]

Hello Santha,
Again, congratulations on your persistence in working through the subtle points of this experiment.

Also, thank you for the link to the journal article on the antifungal effects of Tea Tree oil. Those authors report that Tea Tree oil altered the permeability and membrane fluidity of the fungus Candida albicans. However, those authors did not study the antifungal activity of Tea Tree oil. That is, they did not measure how effective Tea Tree oil was in stopping the growth of fungus. This might seem like a subtle difference, but your project is studying the effect of Tea Tree oil in stopping the growth of yeast. So you need to find a journal article that reports the antifungal activity of Tea Tree oil so you will be able to use the reported minimum concentration of Tea Tree oil which killed their target fungus ("minimum lethal concentration (MLC)" or "minimum inhibitory concentration (MIC)."

To answer the second part of your question - assuming that you want to get a 1% concentration from the 100% concentration, you would want to do two ten-fold dilutions (sometimes called "serial dilutions"), use step 9 (b) (i) for the first ten-fold, and then repeat that step, using 1/8 tsp of the ten-fold dilution.

But remember, that you may find the journal articles report a lower concentration for the MLC or MIC. You will want to use the lower concentration, which might be as low as 0.25%, or maybe even lower!

Let us know what you find and if you have more questions.

[santha123]

Hi Madeline,

I found this website (http://www.webmd.com/vitamins-supplemen ... ientid=113). It only states the concentration, but not the amount to use.

This website says to use 4-5 drops of the oil (http://www.goodhealthacademy.com/how-to ... il-fungus/)

This website says 1/2 tsp oil + 1/2 cup water (http://homeremediesforlife.com/tea-tree ... infection/)

This website says "the MICs of tea tree oil is between 0.125 and 2%" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3264233/)

This website says "Reported MICs range from 0.12% to 2%" (https://www.drugs.com/npp/tea-tree-oil.html)

This website also says that "fungicidal concentrations generally range from 0.12 to 2%" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1360273/)


***Overall 1% is in the range (which will keep all of my concentrations the same.) So, to make the tea tree oil a 1% concentration, I should add 1/4 tsp oil and 18/8 tsp water. Then follow steps 9b and so on.

Thank you so much in advance,
Santha

[MadelineB]

Hooray Santha,
Congratulations on your persistence and patience!

Since the reported MICs are in a range like .1% to 2%, you might want to pick a high concentration and also a low concentration. I realize that you started out thinking to just test one concentration of the TTO, but test a low and a high fits in with the concentrations of the other antifungals that you are testing. Makes for a more complete experiment!

Best of luck!

[santha123]

Hi Madeline,

So I finally finished my experiment. I calculated how much the yeast rose by measuring it in terms of water. I measured how much water it is equivalent to, but I'm not sure what the numbers mean. I'm unable to send you the picture but here are my data:

I also have an outlier (OH Trial 1), but other than that the amount of water for all 3 trials seem relatively the same. I'm not sure what these numbers means in terms of if the experiment worked or not? Also when I am doing a t-test, I am comparing each other experiment (TH...) with the control and for the test, I'm not sure which sign to use (greater or less)? The numbers for the most part, are greater but I'm not sure in this case if greater is better?

Trial 1(mL)
Control 155
TH 232.5
TL 235
UH 242.5
UL 190
OH 397.5
OL 167.5


Trial 2(mL)
Control 227.5
TH 260
TL 240
UH 215
UL 240
OH 210
OL 145


Trial 3(mL)
Control 230
TH 255
TL 250
UH 235
UL 235
OH 135
OL 135

Thank you,
Santha

[MadelineB]

Hello Santha,
Thank you for posting your data!

Before you start thinking about testing the differences between the different concentrations, I would suggest that you look at the data as 3 replicates for the control, and for each antifungal.

That is, first, look at the 3 replicates for the control: Rep 1: 155, Rep 2: 227.5, Rep 3: 230. There seems to be a bit too much variability across your 3 replicates just for the control. With that much variability in the control, it can be difficult to make sensible comparisons between the antifungal agents and concentrations.

I wonder if the water temperature was the same for each of the replicates? Did you collect the CO2 at the same time for each replicate? I know the project gives a range, but you need to select a time, and use the same time and temperature for each replicate. Did you record the time and the temperature in your lab book for each replicate? Did you make sure that all of the air was out of the water before you began collecting the CO2?

I know that it can be frustrating trying to do each of the steps exactly the same for each of the trials, but do not despair!

I think it would be very useful to repeat the measurements for the control for 3 more times, trying very hard to keep the temperature the same, record your measurements at exactly the same time.

I will see if another expert here at Science Buddies has experience with this project and can offer some more advice.

So don't get discouraged! Science is doing lots of picky details!!

[santha123]

Hi Madeline,

I kept the timer for 30 min for every trial and 110F for every trial. I do see that trial 1 is off but 2 and 3 are relatively similar. The thing that I observed for each trial is that at least 2 are very similar and one is kinda off. It could be that since I have to make the mix for each experiment every time, during that process the water might have cooled down, but even if it did, I reheated it to make sure it was 110F before combining everything.


-Santha

[MadelineB]

Hello Santha,
Thank you for your patience and persistence.

I'm a bit confused when you say "for each trial is that at least 2 are very similar and one is kinda off" ...

Are you looking at the results for each trial? Or the results for each treatment group for the 3 trials?

If you haven't already done this, I suggest you make a table with a row for each treatment group (think of "control" as its own treatment group) and three columns, one column for each trial. You could also make a graph, with the horizontal axis indicating the treatment group and the vertical axis showing the values observed for the 3 trials for each treatment group.

Now think about which results would be expected to be most similar across the 3 trials.

And which group of 3 results will be your baseline, to be used for comparing the effects of treatments?

Here's a link to the section on Data Analysis and Graphs in the Science Buddies Project Guide.
https://www.sciencebuddies.org/science- ... l#overview

That might help you examine your results.

Be sure to let us know if you have more questions.

Madeline

[santha123]

Hi Madeline,

The deadline for my project is approaching very soon and I don't know what the numbers mean. I am doing more trials as you have suggested. I have to write a junior reseach paper and for that I need things like results, conculsion, analysis and ect. Is it better if there is more CO2 in the control or the antifungal treatments?


Thank you,
Santha Rani

[MadelineB]

Hi Santha,
I think that the background and procedures sections of this project will help you to understand the role of CO2 in measuring the antifungal activity. See if you can answer the list of questions given in the background section.

https://www.sciencebuddies.org/science- ... background

https://www.sciencebuddies.org/science- ... #procedure

Remember that your written report is an important part of your science project. You might find ideas for organizing your report from the Science Buddies Project Guide.

https://www.sciencebuddies.org/science- ... ndex.shtml

Let us know if you have more questions.