Hello! We are excited to be writing our first blog! We are in an AP Biology class at the MMSTC. We are doing research and are solving if the body temperature affects how well Penicillin works at slowing down the growth of Escherichia Coli. We originally came up with this idea by thinking about how we get prescribed antibiotics for infections with a high fever but not when the body is at a regular temperature. Therefore, we came up with the idea of comparing what will happen with a fever compared to the body being at a regular temperature and compared to the body being in hypothermia.

Prior to starting the research, we looked into previous scientific research done on the topic. We found peer-reviewed sources provided by Google Scholar. These sources have provided us with the information that as the incubation temperature (body temp) increases, antibiotics work better on slowing the growth of bacteria. We then agreed and hypothesize that as the body temperature increases, the antibiotics will slow the growth better. Our plans for research are to really understand how body temperature affects the efficacy of Penicillin on E. Coli and why body temperature affects it.

Be sure to check back in to see updates on our research!!

We have officially completed pre-trials! There were some rough patches, but overall they went pretty smoothly. The first step in the process was to pour the nutrient agar into the petri dishes. While that was cooling, we poured 4 mL of water in a test tube. We then lit the Bunsen burner used to sterilize the transfer loop. You know this is working when the end glows red in the fire of the Bunsen burner. After being sterilized, the transfer loop was used to move a little of the E. coli from the starter plate to the 4 mL of water in the test tubes. Next we poured the E. coli water into the petri dish. We had to make sure to tilt the petri dish side to side in order to move the E. coli water around so the bottom is completely covered in bacteria. The excess water in the petri dish was then poured out in the sink. After that, we sterilized the tweezers using the Bunsen burner. The tweezers were then used to drop 3 penicillin disks in the petri dish. Light pressure was used to press the penicillin disks into the bottom of the petri dish to ensure it does not slide around. The petri dishes were then flipped over and placed into the correct incubator.

After 24 hours, it was time to collect the data. How we did this is by measuring the Zone of Inhibition around the penicillin disks. Basically what this means is we measure the circle around the penicillin disks that have no bacteria. A circle was drawn around the zone of inhibition. Inside this circle, 3 rays were drawn and measured. The average of the 3 rays was then found. This was repeated for every penicillin disk in the petri dish, and each penicillin disk counted for it's own trial (one petri dish counts as 3 trials not 1).

Many things went well and many did not. The first thing that went well is that we could see that the zone of inhibition is different among the different incubators. This was the first sign that this experiment might work. Another thing that went good was that we were able to get a rhythm down into how we were setting up the petri dishes. Megan has been transferring the E.coli to the petri dishes and Liz has been pouring the agar and placing the penicillin disks on the Petri dishes with E. coli. Some things that went bad were that if the petri dishes were not flipped over in the incubator, all the moisture in the petri dish went where the E. coli and penicillin disks were. This created a big mess and completely ruined the trials that we had done. Another was that a couple of times E. coli did not end up in the Petri dish. This could have happened in multiple ways. The first way was that the test tube was missed when putting E. coli in it. We could of also accidentally poured 2 test tubes with E. coli in the same petri dish.

In order to produce readable and accurate data, we have had to make some adjustments. Our first major adjustment was during pre-trials when we decided that using 40 degrees Celsius would be better than 39 for our highest incubation temperature. We also decided during pre-trials to use a set measurement of 4 mm of water for inoculating our Petri dishes with E. coli. In order to continue producing good data, we plan on keeping every day consistent, and frequently check each incubator temperature. Setting up our trials the same way every day will keep our data consistent, and making sure each incubator is at the correct temperature will keep our data accurate.

The first submission of the entire research paper has been submitted!! "That's CRAZY to me."-Estapa. Since you've been following along from the beginning, let's talk about what the experiment has found. After finishing the experiment, the data was analyzed. The overall average diameter of the zone of inhibitions (see above for a reminder of what this means) were measured as the following: the 35 degree incubator has an average of 10.91, the 37 degree incubator by the has an average of 21.89, and the 40 degree incubator has an average of 11.33. These can be seen below.

As you can be seen above, the 37 degree incubator mean lies well above the 35 and 40 degree ones. Therefore, 3 Two-sample t tests were completed. Don't worry we won't go into too many details, you will just have to wait to learn that next year. However, these tests confirmed that there WAS a significant difference between the 35 and 37 degree incubator, and another significant difference between the 37 and 40 degree incubators. The third test confirmed that there was NO significant difference between the 35 and 40 degree incubators. These significant results allowed us to conclude that the 37 degree incubator was significantly higher than the others. In the real world, this would mean the at a body temperature of 37 degrees Celsius, also commonly known as normal body temperature, is where antibiotics would work best at slowing the growth of bacteria.

These results contradict the work done previously by professionals. One was by Rashed Noor where he found that E. coli would grow best when incubated at a temperature of 35°C and 37°C compared to 40°C. This could be interpreted that the zone of inhibition around the penicillin disks from the 40°C incubator would be greatest. Another experiment tit contradicts is one from the School of Life and Health Sciences in Whiteland's College at the University of Roehampton in London. They tested the interaction of antibiotics mixing and temperature. They found that although the antibiotics showed increased cooperation, the overall temperature increases the antibiotic efficacy for single antibiotics.

Advice for you:

Research can be stressful and difficult and you may be dreading it for next semester, but you won't be alone, and you won't have to do it all at once. Because research is a semester long project, you will be able to take it step by step with the guidence of your teachers and peers. It's also nice that you have a partner for this project, but remember, you wont be able to have the same partner twice for all four years, and if you dont help them out and carry your weight throughout the semester, you will find it difficult to find a partner for the upcoming years. To make starting your research a little easier, try exploring topics early. They dont have to be super complex and confusing, and you dont need any prior knowlege to have a few ideas. Finding a topic early can be hard and stressful, so exploring early can lighten the load. As you complete your trials, try to remember that the results you get in your experiment may not match the information that you have researched. Many experiments end up like this and it is actually more common than you would think, so don't be worried if you get unexpected results. Make sure you also take the advice from you teachers! They have done this for years and know what would be best for your project.

We have reached the end of our research and blog! We just wanted to thank everyone for reading this and following along with our research and updates! A special shoutout to Mr. Estapa, Mrs. Gravel, and Mrs. Tallman for making this possible for us and for their guidence along the way. We also want to wish good luck for students reading this with research next semester and upcoming years!