Over the past couple weeks we have all been working to narrowed down our research topics and ideas into a feasible project. Hannah, Bryce and I have decided to work with the deadly brain eating amoeba, Naegleria fowleri. This summer there was a reported case of this amoeba infecting someone at the Whitewater Center in Charlotte, North Carolina, proving this amoeba lives in North Carolina. Although It is extremely rare for this amoeba to infect humans (under forty cases in the US over the past ten years) it is very fatal and causes death between four and seven days after symptoms appear. The presence of this amoeba in Western North Carolina has not been previously reported. The purpose of our project is to determine whether or not this amoeba is living in our local waters. We will be doing this two different ways, one of which is the screening for antibodies to the genus Naegleria in beaver blood. This will be done by adding extracted blood from locally trapped beavers on the French Broad to a culture of the amoeba N. lovaniensis, which is closely related to N. fowleri. If the amoeba is lysed (broken apart) when the blood is added then we can conclude that there are antibodies to the amoeba present. Beaver blood will be donated to the program and no beavers will be harmed for the purpose of this project.
The next step in our project will be to test the bioremediation of our fungi T. asperellum and M. fragilis. If you remember from my last blog we conducted a preliminary test by measuring the precipitate left but this is not the most accurate and its main purpose was to compare it to the other fungi we tested. This new way of measuring will allow us to use smaller amounts of metals and it will hopefully be very accurate. The florescent in presence of light will glow green and in the presence of a few heavy metals the green glow will become less strong. Using a spectrophotometer, which measures fluorescence at a certain wavelength, we can measure the intensity fo the fluorescence, therefore telling us the concentration of heavy metals present in the water.
In our project, we are testing the bioabsorption ability of four fungi. Biosorption is the use of biomass, in this case fungi, to absorb toxins from a solution. In our study, the toxin will be heavy metals which include lead and iron. Before we started our actual project we needed to see if our fungi would absorb the toxin while it was dead, so we used a precipitate test to determine this. We added lead to water then adding the dead fungi. Nest we let it sit and hopefully absorb the lead dissolved in the water. After this, we took a small sample of water and added Potassium Iodide because it will react with the lead, form a precipitate, and turn a bright yellow. As soon as we added the potassium iodide we could clearly see that the fungi had absorbed a majority of the lead.
Ryu and I have been working hard to align the final steps of our project. We made twenty four containers containing sterile dirt. We are then going to put different measurements of lead in each of them. After that we are going to introduce the two species of fungi that we found at Duke Energy. If the fungi grow we will then add earthworms into the soil to help them bioremediate the lead out of the soil.
The worm part of our project is just getting started. The red earthworms just arrived and are ready to be subjected to coal ash filled with heavy metals. The specific earthworm (Eisenia fetida) Ryu and I chose is known to be able to withstand a wide range of environment conditions and fluctuation. The worms will be used for vermiremediation, the process in which earthworms remove toxins from the soil through bioacumulation, of the heavy metals in the coal ash.