Amoeba on your mind? We hope not!


hannah
After the recent media attention Naegleria fowleri, better known as “the brain eating amoeba” has become a serious concern for people around the world. N. fowleri is an amoeba that is found in soil and waterways (natural and man made) around the globe. The amoeba enters the body through the nasal cavity when water is forced into the nose and then moves into the brain. When the amoeba does not find bacteria to feed on it begins eating the brain. N. fowleri are thermophilic amoeba, which means they thrive in warmer waters. Since the amoeba prefer warmer waters previous scientists have investigated the presence of N. fowleri in thermally polluted lakes. In our project we are going to investigate local lakes (2 thermally polluted, 2 not) for the presence of N. fowleri. The two thermally polluted lakes we will test are Lake Keowee and Lake Julian. The two non-thermally polluted lakes we will test will be Lake Jocassee and Fawn Lake. Right now we are working on perfecting PCR and gel electrophoresis (we will test for the presence of N. fowleri DNA) methods before we take our samples from the lakes. We are also waiting on a positive control, already identified N. Fowleri DNA, for our experiment which has proven to be a lot harder and more expensive than we thought. We are also still evaluating the risks of our projects. We believe we have eliminated all of them by not culturing the N. fowleri and just work with it’s DNA. We are all excited and also nervous to see if N. fowleri will be found in our local lakes!hannah1b

Testing Kudzu bug attractions to plant volatiles

Kudzu covers over 7,400,000 acres in the southeastern United States. It is covering most of our state, and national forests, and its destroying wanted vegetation. Because of this problem, we need to get rid of Kudzu.
Our project is to test the levels of E-2- Hexenal of different ages of the kudzu and also test the preferences between kudzu and soybeans of the Megacopta Cribraria (kudzu bug). We think that the bug will be more attracted to the older kudzu and the older soybeans.
By testing the amounts of E-2- Hexenalwe hope to reduce the amount of E-2 hexenal in the soybeans over all to make the bugs less attracted to them. This will save farmers money, so they do not lose as many crops. Forgetting rid of the Kudzu we hope to increaselevels of E-2- Hexenal so they will be more attracted to it and decrease the amount of kudzu in the US.lilly

Rippin’ it in the River

img_3121It has been a couple of weeks since my partner and I last collected T. tubifex samples from the Davidson River. After a “fishy” attempt to collect these oligochaetes from various sections of the Davidson River via the “Coon Tree Picnic Area”, we found that we just weren’t having any luck. So, after we decided that we couldn’t feel our legs from the cold water anymore, we drove down to the local Pisgah Fish Hatchery — in the hopes that someone would be able to give us an idea of where we might find a good sampling site to collect some T. tubifex. Luckily for us, we managed to find an employee, who very kindly described which access points the hatchery typically finds T. tubifex worms on a daily basis. So my partner and I went down through a small patch of woods, and found one of the drainage pipes that connects from the racing wells (where the hatchery fish are kept), to the Davidson River. As expected, we found numerous tubifex worms! Unfortunately, we must not have planned on any success, seeing as though we did not have a tool that could be used to efficiently collect the worms to take back to the lab. Luckily for John and I, we absolutely love digging through fish waste (who doesn’t?), so we just dove right mattin…using sticks that we broke off of trees to pick up some clusters of the worms. They were everywhere! Thousands — likely millions of worms were all gathered in one small area of the Davidson. By 5:45, my partner and I decided it was time to go home, but we did not leave unsuccessful. John took the worms back to his place to sort through them, and the next day I worked on some DNA extraction.

Bean Weevils + Kudzu = Oil?

nicoleThis year BAJONI is down to 2 members. Bain and I are screening the B. ademptus (a bean weevil specific to kudzu seeds) and fungus isolated from the outside of the kudzu seeds for the presence of lipases and proteases. Lipases are enzymes that break down oil and proteases are enzymes that break down proteins. If the fungus or bean weevils test positive for lipases, proteases, or both, then they could later be used in enzymatic oil extraction and produce better, more efficient oil to be used as a biodiesel.

Beaver eight Beaver nine STOP!! its Beaver TIME

aidanOver 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.

Teamwork Makes the Dream Work!

Lauren 2One of the best things about working in the TIME lab is being able to witness a variety of science projects firsthand. Instead of only learning more about your area of research, you have the opportunity to learn about nine other drastically different areas of science. While these projects are very different, they often have overlap which creates a cohesiveness within the classroom.

Brella experienced this firsthand in the recent weeks. As we read more literature on consortium culturing in relation to biofuel, we kept coming across a common fungus- Trichoderma. As we began discussing this more in class, our neighboring team informed us that they had running cultures of this fungus from last year. We were so excited that they were willing to share their cultures with us so we could pursue our project in a new direction!

The competition is always fierce in the TIME classroom, but we also look out for each other. The collaboration that results from working side by side is irreplaceable experience for us and results in great additions to our projects.

And you may….. Begin!

DSCN0087Sarah and I have finally started our testing on the effects of music on cognitive processes and it has been going somewhat smoothly. We have had plenty of bumps in the road thus far, but it is getting easier as we go along. I will say, the three minutes each test takes is the longest part of the process for us and the hardest. The grading is a close second, and getting forms back from people is in third place.  The faces of our participants when we say to put their pencils down is probably the best part, or at least my favorite. Their faces are full of worry, relief, or panic as the timer stops and papers are collected. The reactions of the participants to the music is great to see, whether it is the neutral song or their favorite. Some of the participants love the music and smile and move their head to the beat, and others hate it and frown or get restless in their seat, clearly wanting to make it stop. The relief of being able to say that they’re done with all of the tests is amazing. Well, for us it is. Some of the participants get nervous or worried about how they did even though no one will know who did how well on each test.

I found a BUGGG

I have been “scarifing” Kudzu seeds for several days now. Scarifing is another word for cutting open the seed hulls so that they will germinate. Every other seed or so would be very fat and brittle. The insides would also be juicy and yellow. Carver 2It occurred to me that the yellow juicy stuff was guts of some sort of critter! CarverYes, there is a little yellow larvae of sorts, within the majority of seeds harvested from local Kudzu. So far, I have not found any information on what these little fellers are and why they are in my seeds! Next week I will send a PCR to the gene bank and hopefully Identify what these larvae are.

Lead removing fungi??

AidanIn 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.

DNS (All the kids are doing it these days!)

Don’t worry; this is not some new illegal substance!  Rather, it is an experiment that will be key to the BRELLA project.  As my partners Bryce and Lauren already explained to you, our project took a little longer to get cranking than usual.  Now, after several, SEVERAL attempts at project designs, we are ready to start our preliminary experiments.

DNS stands for dinitrosalicylic acid, a fairly common chemical which is also known to produce a color change when it bonds with glucose.  (This is important because, if you recall from Bryce’s blog, when you are making ethanol, the enzymes break the substrate into glucose molecules.  These are then fermented into ethanol.)  We will be performing an assay using DNS to see if enzymes are being produced by both our consortium and the individual microbes.

Enzymes will be isolated from the broth in which the organisms are growing in.  These isolates will be placed on a piece of filter paper and given time to degrade the paper.  Then DNS will be added and will result in a color change when Elizait bonds with the glucose molecules.  This color change will indicate that the proteins have broken the bonds of the filter paper into straight glucose.  This assay screens for “reducing sugars,” basically meaning that if glucose is present, enzymes had to have been present.  Tomorrow, we will be using performing a positive control test in which there are no enzymes, but instead we just add the DNS to straight glucose.  This will aid us in measuring the next tests.  Stay tuned for data!