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Antibodies!

Today is our last week in lab, my oh my how the time flew! Today we did antibody testing, which was really cool! 

First we labeled our well. There were 3 positive wells, 3 negative, 3 sample #22 and 3 sample #37. 

 We then proceeded to do the experiment. We first put the purified antigen into each of the wells, and then waited 5 minutes for it to bind to the wells. Then we washed the wells by using a buffer. We then put the positive control in to the three "+" wells, put the negative control into the three "-" wells, put sample "22" into the wells labeled 22, and the sample "37" into the well labeled 37. We then waited 5 minutes. 

We then proceeded to wash the wells again with the buffer. After that we added the secondary antibody to all 12 wells, then waited 5 minutes. We washed the wells two times with the buffer, and then proceeded to add the enzyme substrate to all 12 wells. Our results indicated that the positive and the sample #22 turned blue! This means the secondary antibody is attached to an enzyme (HRP) that chemically changes the enzyme substrate. 

Here's our results!:

We also prepared another experiment today. This one involved a technique that has to do with immunology. We made little wells in the petri dish containing agar. We added one solution to each well. These solutions were Bovine Albumin, Goat Anti-horse Albumin, Goat Anti-Bovine Albumin, and Goat anti-swine Albumin. We then placed our dish in a room temperature incubator. We'll see our results on thursday! 

And Our Unknown Bacteria is......

I can't believe it!! Today we worked out what our bacteria is. It's so hard to believe that the semester is almost over. I'll tell you what unknown H is a little later.

As we looked at our antibiotic results we had to measure the radius around the disk to see how well that particular antibiotic worked against our unknown bacteria. Once we know the radius then you get the diameter. With the diameter we looked at a list of diameters for the antibiotics to see how far away the bacterial growth had to be in order for the bacteria to be considered effected by it. 

The most effective against bacterial growth was Erythromycin, Penicillin, and Novobiocin. This would mean that we would probably give penicillin as a first choice drug and it would attack the cell wall. If our patient was allergic though to Penicillin and the other semi-synthetic product then we would like to give either Erythromycin which attacks the function of the ribosomes within the cell or Novobiocin which attacks the nucleic acid synthesis. 

Next we tried to figure out this mysterious bacteria. Dr. P. had us do our gram staining again because we were really unsure if the bacteria was positive of negative. This time he said our stain was beautiful!! And we confirmed that our sample is gram-positive cocci! 

Sorry, I don't have a better picture. Anyway, after following many charts we came up with the name of our bacteria. And the name is...

**dramatic music**

MICROCOCCUS LUTEUS

We were both really happy to get it right. It took lots of help from Dr. P. but we made it!

The yogurt Dr. P. made was also done. The kind in which he boiled the milk first tasted the best.

The blue marked cup is the non-heated milk and the red is the heated milk

Next week is Thanksgiving so we can't run any test on Tuesday. I think we are going to watch a video for lab. I can't wait for break!! 

HAPPY

THANKSGIVING!!

Antibiotics Here We Come

Today we applied five different antibiotics to a spread plate of our unknown H bacteria. We used all four from the first picture and we choose to do Penicillin as our fifth one. We are trying to see if our bacteria is resistant to any of them. making the plate is actually pretty fun. :)

All the antibiotics are on small pads that we place on the spread plate in different quadrants so that we know which antibiotics work and which ones don't.

As an extension from last Thursday we swabbed the same cell phone and tested to see if 70% alcohol  would inhibit the growth of the bacteria better then the 68% alcohol. Well, as usual with all our tests it was uninteresting and neither the 68% or the 70% worked against the growth of the bacteria.

70% is on the bottom half and 60% is on the top

Also at the end of class Dr. P. showed us how to make yogurt! First, he filled two cups with milk. Then in one he put a small spoonful of greek yogurt. In the second cup he first heated it up, let it cool down and then added the yogurt. He then put it in a small incubator and we will see the result next class.
We are nearing the end so the pressure is on to figure out what our unknown bacteria is! Maybe we can do it Thursday.

It's Not as Clean as You Think!

Today we looked at our plate of the bacteria we collected from our classmate's phone. We found that the best means of prevention of bacteria were bleach,        hand soap and Lysol wipes. The LOL hand antibacterial, Lysol spray and mouthwash were not as effective.  

Here's another picture of our test results from the phone's bacteria: gross! 

We also got looked at the test results from the water treatment via the UV light. These results were suprossing to us becasue the water that was treated only once with the UV ligt looked like it had the most bactiera killed. The water that was treated a second time actually had more bacteria than the water that was only treated once. 

Here's a picture from the bacteria in the water: 

 Starting from right to left: Water after 2 treatments, water after 1 treatment, water before treatment.

After we looked at the water tests, we took our bacteria out of the anaerobic chamber. 

We discovered our unknown bacteria is an aerobic bacteria because it did not grow in the anaerobic chamber

Towards the end of lab we did one last experiment. When thinking about the bacteria that grew on the phone and realized that the antibacterial gel LOL only contained %68 ethanol. We proposed that the bacteria would have been killed if it was treated with 70% Ethanol. We inoculated the bacteria in a nutrient agar and then separated the dish into quadrants. One quadrant contained the LOL gel and the other contained 70% Ethanol. We then incubated the bacteria at 37*C. We'll se what happens next week! 

Did an independent experiment to check and see if there was a difference between 68 or 70% Ethanol in the growth of the phone bacteria 

Nose Goes

The first thing we did today was get our bacteria that we had swabbed from our nose. We looked at it under the light, it was pretty gross! 

After we spent some time looking at our bacteria, we did some disinfecting tests. The first thing we did was swab our classmate's cell phone and inoculated it in a nutrient agar plate.

We then took samples of different disinfectants and placed it on our new sample to see if it would grow. We used a bunsen burner to sterilize our forceps each time we obtained a new disinfectant.

We divided our nutrient agar into four quadrants and placed a different type of disinfectant in each one. 

Our disinfectants were tooth paste, mouthwash, bleach, and hand sanitizer. We incubated our sample at room temperature (25*C)

 After that we made a streak plate of our unknown bacteria. We are testing to determine if it is an obligate anaerobe bacteria. We took the bacteria to an anaerobic chamber. We incubated the chamber at 37*

We all then took a sample of bacteria that we had obtained over the semester, and placed them in a beaker of water. The idea behind this was to mirror a pond or body of water that has a lot of bacteria in it. Our goal was to make this water drinkable. We did this by using UV light to purify the water.

We started this experiment with Juliette swabbing a sample of the bacteria water on an agar plate. We then used a UV light to kill the bacteria. 

After this Juliette swabbed the water once again on an agar plate to test and see if the water had been successfully purified. We then treated the water with UV light and took another sample of the water. 

We put the plates in the incubator. We'll see what grows by Thursday!

Nursing Practice!!

We are now moving into more clinic applications of microbiology. Today we practiced doing nose swabs! Yeah, I know it doesn't sound all the fun to some people, but we all had a blast. I swabbed my partner's nose and we did a spread plate of the sample. We will see what grew on Tuesday.

The process was simple. First, we had to wet the swab in a saline solution so that bacteria would stick to the swab

The next step was swabbing. It wasn't as awkward as it looks. Not my best picture. Oh the things we do for school. Well, we'll see you next week!! :)

Field trip!!!

Tuesday was our field trip day. Dr. P. took us to the waste water treatment plant where we got to see and learn how waste water is safely treated before entering the river. The facility manager gave us all the tour. He took us through the facility along the path the water travels from where it enters to where it exists into the river. He told us all about the types of microbes that are use in the process (All the microbes are aerobic). It was amazing to see the details. It has definitely given me a great appreciation for the job that these men do. They are so smart!! He was telling us all about the chemical reactions that these microbes have and why this one is better than that one because it can be used up in the reaction so it never is released back into the environment. He completely lost me at some points, but it was really amazing. These men are so smart and I can thank them enough for the job they do.

I forgot my camera when we went so I'm sorry that there's no pictures.

Interesting...

So we have more results and possibly a contradictory one. Lets start with our Thioglycollate test which shows us the type of environment our bacteria can grow in. In our test tube, the bacteria grew all throughout the tube! This told us that our bacteria H has facilitated growth meaning it can grow with or without oxygen in the environment.

So it's hard to see, but there is bacteria all throughout the tube

The school photographer also came to our class in order to take some picture. It was a little awkward at times to say the least.

Anyway, moving on to other things.

Next we looked at our last enzyme test which was on DNA agar. We had to add hydrochloric acid to the plate in order to see the result.

Our bacteria was as you can guess.........negative. For each of these tests we split the plates in half so our bacteria is the yellow one and it should be growing in the shape of J and M. With this test being negative, it tells us that our bacteria does not have the enzyme used to break down DNA.

Now we will look at our last five tests. These were all on selective or differential media. They will tell us where our bacteria can grow.



First is blood agar. Our bacteria grew, but did not have the ability to lyse red blood cells (in other word break them down).

The next one was EMB (eosin methylene blue). This one was negative as it was lacking any growth what so ever. Some of these tests we don't know yet what the results mean, but Dr. P. assured us that we will be learning about it in lecture soon.

The Mannitol Salt agar was then under scrutiny.  Since this agar is a salt most bacteria won't grow here because of the hypertonic state. The salt will draw all the water out of the bacteria and it will die. This is why pickled foods last so long because lots os bacteria cannot grow in the salty environment. Our bacteria is the one above the line and it did not survive.

Our fourth one is the MacConkey agar. This one is suppose to inhibit gram-positive bacteria. Other then that all we know it that we had no growth. What this result means, again we do not know yet.

The fifth and final test to look at was the PEA test (Phenylethyl Alcohol Agar). This one was interesting because this agar inhibits or slows gram-negative growth. We beleaved that from our gram stain that our bacteria was gram-negative. The problem though is that we has growth on this plate!!

The amount is very small so it could be that our mystery H bacteria was still able in some way to grow just a tiny bit, but it still makes us question if our original thoughts was wrong. Next week we may repeat the gram stain and see if we still come up with the same results. Until then, have a safe weekend!!

More samples and Strep throat swabbing practice

We are now half way through our tests for our mystery bacteria H. Today we looked at the streak plate we did last week. The bacteria grew on the surface so it must be aerobic. To see if bacteria H has any peroxisomes we poured hydrogen peroxide on the surface of the plate. Our's was slow to bubble, but after a few seconds it started to react. This told us that our bacteria is definitely aerobic because the catalyst needs to be present in order for there to be oxygen for the bacteria to grow.

We shared our plate with another group so our bacteria is the yellow one.

 We also inoculated seven more tests. Five of them are called differential media tests, one was to test for the hydrolytic enzymes for DNA, and one was to test the anaerobic or aerobic type of bacteria.

Anaerobic or Aerobic?
After inoculating 

Anaerobic or Aerobic?
Before inoculating 

The five test we started on different media were:
EMB, Blood, PE, Mannitol Salt Agar, and MacConkey Agar

We got to be all official and use the disposable tips to inoculate!!
So the last thing we did was practice swabbing each other's throats for strep. We didn't test them all, but we got to practice the technique. It was super fun. hardly anyone gagged so I hope we did it right! Anyway, on Thursday we will see if our bacteria has decided to be interesting or not. We shall see.

Urea! Things are looking up.

Thursday, October 17th

WE FINALLY HAVE A POSITIVE TEST!!!!

Our test for Urea came back positive. Because of this we can conclude that our bacteria can hydrolyze urea. 

Not surprising however, the rest of our tests came back negative. Here's a couple of examples of our negative tests:

                   Nitrate:                                                                                                 Indole:

After we checked our test results we completed an oxidase test. 

Our results came back negative: