Pre-Presentation Jitter's

I am not completely sure if my title is grammatically correct, but then again that is why I am a science major and not English. Never really was a strong subject for me. Well, the conference is getting closer (Monday the 4th to be exact) and I am going to be completely honest, I am getting more and more nervous. I have officially completed my poster COMPLETELY today. Matt and I figured out a way to fit everything on the poster, even with the very limited space we were given. I was even able to fit a few, small, pictures in between the madness of a poster I created.    

Below is the completed poster that is going to be presented at the conference. I am actually very excited to see the different projects that are going to be presented. I was able to present last year at the conference held at Estrella Mountain Community College, and I have a feeling that this conference at ASU is going to be a lot more intriguing. At least I am hoping. 

As you can tell there was a lot of information squished into a 3x3 poster. It was somewhat difficult but it is done and I am happy to say I am ready for the printer :)
After a long semester of tests, bacteria and more tests, it is quite the accomplishment to say that I have completed my project and I am ready to demonstrate my accomplishments too the many people attending this conference.

I don’t know if we are going to practice presentation before Monday, like we were able to do last year, but no matter what I have been putting together little notes for myself that will help me when presenting. I know my project like the back of my hand, which makes me feel a little bit more comfortable with presentation. Well, I guess that is it for now.

Oops!... But on the brighter side...

So I regret to admit that it COMPLETELY forgot to post last Thursday. It was a mistake that I don't intend to let happen again. Although I was a little disappointed in my forgetful mind I am very pleased and, I have to admit pretty excited, to inform you guys that I have completed my poster with both dichotomous key's. I finished with enough time to edit and practice it to a point were it can be flawless. Okay, maybe not "flawless" but pretty close.

While I was doing the poster I didn't realize how difficult it was going to be due to the size of the poster. Well I should clarify that it was more frustrating rather than difficult. The size requirements for the poster is a 3x3 finished poster. My dichotomous key's alone were 3x3 (not really but kind of close) and I had to figure out a way to fit both dichotomous key's plus my data, an abstract and so much more. It was like playing Tetris while creating a poster all at the same time.

I am not completely satisfied with my poster as is but since I am done ahead of schedule I will have the ability to adjust as I feel necessary. The wonderful perk of finishing early.

    This is the gram negative dichotomous key which is only on here curtsy of Josh. I have two dichotomous key's as the end product of my project and it is quite similar to the one above. Well, I guess that's all I really have at the moment to inform you guys about.  

Hmmmm....

Well to start off this weeks post, let me tell you how close I am getting to completing my second dichotomous key. I did run into another little problem yesterday with two of my bacteria's results. With the results I have, currently, it looks like Salmonella and S. Marcescens have pretty much all the same results. That is a problem for me because I don't know how to show identification between the two. 

What we decided to do was, attempt growing on a media that I couldn't do at first (because we did not have the media until this week) which is blood agar. Blood agar basically shows the bacteria's capability to breakdown hemoglobin in blood. 

I inoculated all 15 bacteria's yesterday and tried to read them as soon as I got here today.

 

 To sum it all up, my results were unreadable because I was not aware that they had to be inoculated in a candle jar. The picture on the left was one of my more clear results that we were able to read, somewhat, but the others just turned completely brown. Which, I learned, means the blood in the plate had began to oxidase, or had too much exposure to oxygen. 

An example of that could be; the color of blood before it hits the surface (oxygen), then it turns a darker, almost black, color. Basically the same idea. 

The picture to the right is the correct way I should have inoculated the plates. The plates are put in a jar with an ignited candle on the inside as well (thus the name "candle jar"). This is to remove the oxygen present in the jar.

Then, as extra security, the spaced between the lid (of the jar) and the lip, is parafilmed.   

For some of you that have read my blog, you will see that this is not my first problem or mistake. So, just like the mistakes before I will continue until I find the results I am looking for. I am not too worried about it now because I have a little time to fix LITTLE errors. I do want to get started on my poster for the presentation as soon as possible so it can be done without flaws and I can have enough time to practice what I am going to say. But it looks like once I work through this little problem I can get started on my poster. =)

Ahhhh!!

Hello, well let's see what is new to talk about.
First off, I did get a response back from the ASU Conference, so it looks like I will be presenting my project on March 4^th. I am going to admit, I am somewhat scared and excited to have the opportunity to present at ASU. Let’s see how it goes.

As for my project I have hit a sort of wall. I realized that some of my test might have been misread and so I have decided to take the safe route and just redo some of the test in question.

What I found pretty interesting was the results of Strept. Mutans, which was one of the bacteria’s, in a glucose fermentation broth. Glucose FB is a broth that when inoculated with certain bacteria’s, will determine if an acid or gas (or both) is produced.

In the image to your right, you will see how the glucose broth originally looks like.

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If you can tell the broth is a sort of orange color and there is a small test tube in the larger test tube. Those are called duram test tubes and the purpose of those tests tubes is the read whether gas was produced from the bacteria. If the bacterium is positive for gas production then a bubble will appear on the top of the duram tube. The reading that I found interesting was of this image below.

  

If you can see there is a growth, or build up, in the duram tube where the gas bubble should be. This is something that I have never seen even with the other 14 bacteria’s that went through the same test. I did manage to ask several people about the growth but nobody knew what it could be. So what I then proceeded to do was basically redo the glucose test for Strept. Mutans. This will help me determine if this is usual for Strept. Mutans in glucose fermentation broths or if it could possibly be a contamination.

Ohh and just cause I want to share a little of my insanity, not really but almost there, I have uploaded another picture of the choas that surounds me currently. Thus the reason for the title Ahhh...

Update 01/23/2013

Well I am still working on my bacteria identification project. I am currently in the process of creating a dichotomous key that will make the identification of bacteria simpler, but I have also been able to determine what tests give the best and clearest results in the end. I have encountered a few bumps along the road but all in all I feel my dichotomous key is coming along just well. I am aiming to have a completed one by next week so I can have Matt edit it and I can move on from there. I have already entered my application for the MGE Conference at ASU, now all there is to do is wait for a response. The next step after the completion of the dichotomous key would be to begin on my poster. I have only done one other poster before this one but I am confident that I will be able to produce a more than excellent poster. I don’t have many pictures of what I am doing but I did take the liberty to take a picture of the mess I have created in the process of working.  

As messy as my area is beginning to look, I can only imagine how my brain is starting to look like =(

Guess who's back?..

Hello my fellow peers. I am back from a horrible week of sickness. Now that I am over that its back to the real life messing with all my bacterias. Well I think I last left off when I was beginning the new set of bacteria. To start, I looked to see what other bacteria we had in our lab that I still hadn't done. That kind of gave me a rough list of the rest that needed to be tested. This list, I must say, was a little frighting at first. It was a long list consisting of 19 bacteria all together (or 10 more than what I already have completed). I began to make a whole new stock for my use only with the remaining 10 bacteria.

My stock bacteria, which is just the concentrated bacteria grown in TSB and THI solutions.

 I also remade the previous nine that I had already done most of the tests on. I began all the test by first growing my stock on TSA plates so I can have a active bacteria when doing these reaction tests.

This is one of my plates that I grew on TSA. I choose this one just because it looked the coolest. This is what I grew all of my bacterias on and with the colonies grown on these plates I was able to run multiple test from them.

The first problem I ran into was how 5 of my TSA plates just refused to grow. That is a big problem for me because that was how I was going to do all of my tests. Not trying to throw in the towel quite so early, I then proceed to try and grow these 5 different plates in candle jars. From talking with the pro of micro (Jenn & Matt) told me this was probably the only way these bacterias were going to be able to grow on TSA. I honestly didn't think it was going to be that hard, I mean I have watched Jenn do this whole process about 50 times, how hard could it be, right?

WRONG! Ohh so wrong..

A process that seem so easy became to be one of the more difficult. Okay, maybe I am exaggerating just a bit, but it was more difficult then I had first pictured in my head. In order to do these you streak your TSA plates as normal but instead of incubating them bare you put them in these big pickle jar looking things. Once they are in there, you must add a candle that has been ignited, out on the cap and parafilm around the cap. Seems easy right? No, I would light the candle and try and drop it on top of the plates (in the jar) as carefully as I possibly could but it kept either falling to the side or turning off before I could even get to the "putting on the top" part. With frustrations beginning to over take my patience I tried again and got it.

To my disappointment, the whole candle jar only helped one of my bacterias grow. So with time running thin with this project, I decided to move on with the bacterias I did have (which was a good 6 other bacterias). Which seemed to be so much easy to do the second time around, now that I knew what I was doing, oh the joy of doing this several times before. I have gotten through with the majority of my tests and I could have completely finished last week (if I hadn't got sick) but I was able to complete the last 4 test today and will get the results by tomorrow. So do expect a follow up post to this one, soon.

So Many Test Tubes, So Little Time...

I am not getting close to the end of my project. I have completed another four test to the same 9 bacterias, but I have to redo one of the test. Which was caused by, me letting it inoculate for too long. We also decided that the o/f medium (or oxidation fermentation medium) is very unreliable and difficult to read. This has given us the idea of finding a more efficient way of preforming this test. What this test detects is if acid is given off after fermentation or oxidation. This is suppose to allow for a color change to happen in the media, changing it to a yellow rather than the dark green that it began. This is due to a lowered pH after oxidation occurs in the process of inoculation. From my practice doing the test on all the bacterias, I realized that it was very difficult to get a significant amount of bacteria in the test tube, while still trying to follow the directions of how to correctly "stab" the media. This could have been one of the issues because its not given enough bacteria to react to.
The one test I really found interesting was the T.S.I Slants (Triple Sugar Iron Agar). This is another acid test that detects presence of acid and Sodium Thiosulfate (reduced to H2S gas). If positive for the gas the media would change colors from yellow to black (or a very dark red, in my test). The reaction occurs because H2S reacts with ferrous sulfide in order to create the black precipitate that grows mainly from the "butt" of the media in the test tube.

This is an example of a positive reaction for acid from sugar fermentation in the bacteria. What was so interesting about this tube in particular is the fact the the actual media moved that high in the tube. They all started off at the same height and as you can see in the image above, the media completely shifted upward in the tube.

 

These images above are examples of positive reactions for H2S gas. The very first one was the one that I found most fascinating mainly because of the intense color change. Remember, they all started off a yellow color, most similar to the media shown on the far top. 
Well that's pretty much where I am at now, and I just found out that I will doing all these test AGAIN, with 12 different bacteria's. So, wish me luck! :[