3/27/12

Today was not the greatest. I had to measure the width of the stomata for all of my plants and finishing up the density. My powerpoint is almost complete and also a work in progress. (Mostly done)

3/21-22/12

These days were focused and dedicated on working on my powerpoint and paper for our presentation. Not much exciting details for here but my presentation will look great as it's coming together nicely!

Back from Spring Break

Return from spring break. It's been a while since I have been in the lab because of a week off. However, as soon I returned I immediately continued on an experiment that I left off on, and I also started a new experiment today. 

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Bioremediation Tests 

Before the break, I had set up 13 test tubes with 13 different species of bacteria and Crisco oil. I didn't need to include the color indicator tetrazolium because the point of the test was to see if bacteria truly did do something with the oil, and also see if there would be some kind of cloudy appearance. I noticed small spots at the bottom of the test tubes that had the bacteria:

• E. coli
• Pseudomonos fluoroscens
• Salmonella
• Serratia marcescens
• Proteus vulgaris

I wondered why these specific species of bacteria had those results but the other results didn't. I showed Matt and Josh my results and I was told to repeat the experiment but this time add 1 drop of the bacteria rather than a loop, and also try a new oil -- motor oil. I may have to continue with the additional procedures another day because I needed more material like test tubes, and the room I usually work on my experiments was busy. 

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Bacteria Chart
Today I also worked on a chart, and the data that I used for the chart was a bioremediation experiment weeks ago using different species of bacteria, corn oil, and tetrazolium to see which ones turned red/dark pink. The chart consisted of four things:

1. The specie of the bacteria.
2. If there was color change or not. 
3. Gram positive or negative. 
4. Cell shape. 

(Click for better view)

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2/29-3/1-3/7-3/8

Wow, first of all, I apologize for this very late post. I did my work but forgot to post. These days were all about working on my abstract for the Estrella Mountain Community College Student Conference. I had Matt revise it, I re-worded again then worked through it with Matt and submitted it.

I also started working on a table for my project on stomata. I listed all the species of plants by common name and scientific name. I have to add the different densities.

Caffeinated

Today went by pretty fast. I was surprised how quickly time went by today and I did not even get to the second part of my newest experiment yet.

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First, 

I checked the incubator today to see how the DI water and seawater with Crisco oil and motor oil resulted with the bacteria. As of now, there are no obvious change and I recorded "no change". Change probably did occur, but if it did then it was not visible because I did not include tetrazolium (color indicator that determines if bacteria degrades oil). The only new observation that I did notice were very small bubbles (about 4-5 in each plate), but other than that nothing. I may need to wait longer than a day to see what the results were. I returned everything in an incubator and plan to take observations within a lengthy period to compare a day to about a week (more of less). 

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More soda and exercising!

Esperanza asked if she could run a test and I would be a subject which I agreed, but what I had to do was drink a can of soda in about 5 minutes and then wait 20 minutes after drinking it. I couldn't drink it down in 5 minutes, but I did my best to make sure that I finished it and then set the timer of 20 minutes. After, I went to Esperanza and what I had to do was click a button (w/ headphones on) every time I hear a sound. After, I was told to run up to the highest floor of the building using the stairs, and then run back down. I then needed to repeat the hearing test and the results came better than my first attempt. 

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Working with more bacteria and oil

Matt gave me a new experiment which was to apply 2 mL of Crisco oil in 13 tubes, and then apply 13 different species of bacteria. I was curious to see how this would work, or how I would know if the bacteria is working without the addition of tetrazolium (the color indicator). After inoculating the species of bacter to the tubes with oil, I let it set in a test rack at room temperature. 

Petri Dish Experiment

On Tuesday (yesterday) and today, Josh introduced me to a new experiment. For weeks I have been testing to see how bacteria reacted, if possible, to corn oil. After several studies, I found that for the most part all the species of bacteria that I chose ranging from E. coli to Bacillus subtilis. I immediately noticed a sudden change of pink color in the tubes compared to Enterobacter aerogenes. However, it becomes strange that a particular bacteria took days or even a week to show results while these other species of bacteria had five-second effects.

This is when a new experiment arose. This time, I would use six blank petri dishes and then add certain key ingredients.The purpose of this experiment was to apply two different species of bacteria to four petri dishes with distilled water or salt water and oil applied. I had to do some background research to find how to make seawater (my original task), but we did not have the necessary ingredients to make it at the moment. Instead, I found how to make 'seawater'.

Preparation

1. To make table 'seawater', have 35 grams of Kosher salt and apply it to a contained 965g of distilled water.
2. Stir the water and the salt until there are no salt particles present. 
3. Next, apply distilled water into three petri dishes, enough so that there are no open gaps. 
4. Repeat step 3 with seawater. 
5. Apply 5 microlitre of Crisco oil to petri dishes (2 labeled with seawater and 2 labeled distilled). [One petri dish will only contain DI H20 and Seawater to be used as control plates.]
6. Wait for 20 minutes for any spreading of the oil.
7. With a thin ruler, measure the diameter of the round oil shape for each petri dish. 
8. Apply one drop of designated bacteria to the designated plates. 
9. Carefully placed the plates in an incubator and wait for 24 hours. 

*I also replaced Cisco oil with motor oil, but did the exact same steps. 

Results

The ones with Cisco oil: I noticed that the plates with distilled water formed a small circle of only .5 centimeter while the plates containing seawater had more of a spread with the oil and the measure of the diameter was 1 centimeter.

The ones with motor oil: I saw immediate dispersion of the oil into the water. I waited for 20 minutes to see how it would form and then apply results.Since the oil was still segregated, I measured the default length of the diameter of the petri dish which is 14 cm. 

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It has been two days with this experiment. There were some flaws and little success, but I will see tomorrow if the newly added plates containing a couple of bacteria have shown results.

*I also submitted my abstract, but before that I had to continuously check for errors and make sure that it was solid. I was grateful for the feedback and help that I received to make it turn out as it is now.

Today's Color: Pink

Recap: Yesterday I incubated a total of 24 petri dishes. There was a total of 12 different species of bacteria (and a duplicate for each bacteria). I needed to wait 24 hours for the bacteria to incubate to begin the bioremedation experiments. One experiment (experiment 1) would consist of a control tube (just 2 mL of tetrazoilium and 10 drops of corn oil), and other tubes with the same substances but the unique bacteria added in each tube. Another experiment (experiment 2) would consist of just the 2 mL of tetrazolium, and tetrazolium with the unique species of bacteria. 

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Results from incubation of each bacteria

Bacillus Subtilis that has been streaked and incubated for 24 hours. It looks like thick circles that form some kind of worm-like appearance. 

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Experiment 1 and 2

Bacteria: Salmonella enterica 

There was immediate result when adding Salmonella to the tetrazolium and corn oil. Since there are obvious signs of a pink color, it shows that Salmonella is capable of degrading the oil. 

With experiment 1, I tested 10 tubes and one control tube which was just 2mL of tetrazolium and 10 drops of corn oil. 


The types of bacteria used:

• Bacillus subtilis
• E. coli 
• Serratia marcescens 
• Salmonella enterica
• Proteus vulgaris
• Enterococcus faecalis
• Pseudomonas aeroginosa
• Staphylococcus epidermidis 
• Proteus mirabilis  

For the most part, the results showed many of the species of bacteria to take immediate or gradual, but quick effect of changing into a pinkish to dark pink color. Usually, it takes a day or two to take effect (Enterobacter aerogenes), but many of the bacteria tested were much quicker than that. Experiment 2 also was the same, but I excluded corn oil and left it only with tetrazolium and added the same species of bacteria. All same results except P. aeroginosa, E. faecalis and S. epidermidis showed no effect.

[Bacillus subtilis (gram positive) takes immediate effect on just tetrazolium]