Mutant Bacteria Strain?? Antibiotic Test Results and Bacteria J Revealed!!

Mutant Bacteria Strain?? Antibiotic Test Results and Bacteria J Revealed!!

Class: November 21, 2013

A few class ago, Dr. P demonstrated UV radiation use in killing bacteria.  (Remember the light saber??) It supposedly kills EVERYTHING (even spores!), but we wanted to know just how effective the UV light, so we grew three spread plates, one before the UV treatment, one after the first treatment, and another after the second treatment.  You'd think it would be pretty effective after TWO treatments, right? WRONG.  The pre-treatment plate was of course the worst, but there was a pretty gruesome amount of bacteria growing on the first post-treatment plate, and even more on the second!! This made no sense, so we pursued it further and infected some DiH water with this particular bacteria, did the UV light treatment again, and made another spread plate.  Before treating the water, we cleaned the SteriPen tip with alcohol and did a spread plate of that, just to be sure it wasn't an alcohol resistant bacteria on the tip of the pen.  We have the results today, and they are pretty interesting.  There were zero bacterial colonies on the alcohol-sterilized-tip spread plate...

It is a little difficult to see, but take my word for it
there are nada bacteria on this spread plate!

However, there were 3 very small colonies growing after treating the water (it was treated one time).  So, it would seem that we have a bacteria on our hands that is resistant to UV radiation. Rare for sure.  

Last week we created a spread plate of Bacteria J and placed various antibiotic discs on it, which we sectioned off and labeled:

When assessing bacterial effectiveness, we measure the circumference of clear space around the antibiotic disc.  The greater the circumference, the more effective that particular antibiotic is.  Here's our line up:

• Novobiocin- 5.3 cm
• Erythromiocin- 4.6 cm
• Neomyocin- 1.4 cm
• Tetracyclin- 3.5cm 
• Penacillin- 2cm

The winner clearly is Novobiocin, the least effective being Neomyocin.  We conducted this test to further support our current findings about Bacteria J, namely that it is a Gram-positive bacteria.  We know that Gram positive bacteria have peptidoglycin in their cell membrane in large quantities, so different antibiotics are more effective then others, depending on the bacterial structure they target.  Novobiocin is a DNA synthesis inhibitor, which is greatly effective across a broad spectrum of bacteria.  Erythromiocin and Tetracyclin are both cell wall-targeting antibiotics, so it makes sense that they were both fairly effective in killing Bacteria J.  It's awesome when your test results support the rest of your research :)

ATTENTION PLEASE. IMPORTANT ANNOUNCEMENT: MYSTERY BACTERIA REVEALED!

After a semester of careful testing, staining, spreading, microscopic viewing, and being disappointed by almost all the results, we finally nailed down who Bacteria J is! World, meet Micrococcus Roseus: 

Around here, we like to call her Rosie.  Especially Matt.  He's really fond of her.  In order to track down exactly what species of bacteria this was, Dr. P. gave us a few flow charts with various classifications of bacteria on it, that compiled also the test results.  We followed the flow chart for gram-positive coccus bacteria, and almost chose Micrococcus Leteus, but upon further research we found that Micrococcus Roseus was nearly identical in reaction and morphology, except that it is pink, the exact description of our very own Bacteria J :) Team, well done. 

Let's celebrate with a little lab-made yogurt shall we?

It was probably the best yogurt I've ever had.  No joke.  

Mission Impossible Dos: Did we kill the bacteria?Isourbacteriaobligateanaeorobe? What about that lightsaber?

Mission Impossible Dos: Did we kill the bacteria? Is our bacteria obligate anaeorobe? What about that lightsaber?

Class: Thursday, November 14, 2013

Today was exciting! We checked the results of our last lab. In that lab we prepared several tests. In one test we used my phone, swabbed it, and spread it on an agar plate. On the agar plate we placed four disinfectants/antibacterial cleaning agents--2% Lysol spray, Lysol disinfectant spray (in Mandarin orange), foaming antibacterial hand soap, and Lysol disinfectant wipes. The purpose of this test was to prove [or disprove] the antibacterial properties of the cleaning agents.

Did the cleaning agents kill the bacteria that was on my phone? Let's look!

Do you see any bacteria?!

We saw a little bit of bacteria in two of the four quadrants. Do you see them? There's bacterial growth in Lysol (Mandarin orange) spray and the antibacterial hand soap! So now we know neither of those two cleaning agents would be too effective with cleaning my phone.

Next, we checked on our mystery bacteria we put in the air-lock chamber or the anoxalte chamber during our last lab. We were testing our bacteria for growth properties [obligate aerobe/anaeorbe]. Today we revealed that property! What will it be? Can our bacteria grow in an environment without oxygen or does it need oxygen to grow? Let's open up that chamber and see!

We took it out of the chamber and revealed our bacteria! We shared the agar plate with Mary and Jess, so I must clarify, our bacteria is in the top half of the plate. As we can see, there is a little bit of bacterial growth! There we go bacteria J, I knew you had it in you!

Last lab, we made a concoction of all of our bacterium in water and zapped it with  a steripen to test its effectiveness against bacteria. The Steripen uses UV light to kill all bacteria in water making it safe to drink. We swabbed the bacteria infested water before UV exposure, after UV exposure, and after a second round of UV. Let's see if it actually works or not!

Left-Right: BeforeUV, 1st UV, 2nd UV

Well, this didn't make much sense to us! Sure, the "before" plate does, just look at all that bacteria! Gross! The second plate shows a significant difference in the amount of bacteria. The third plate [after 2nd time of UV exposure] shows more bacteria. In theory, there should have been less. We think we may have done something wrong, so we went ahead and prepared a second trial test with the Steripen. Hopefully this time we do it, the results are what they are supposed to be!

2nd Trial-Before UV

2nd Trial-UV exposure

2nd Trial-2nd UV exposure

2nd Trial-1st UV exposure

NOTHING gets in the way of Dr. P from teaching!

We are The Pursuers

We are The Pursuers

The Jolly Truth, as portrayed in the follow pictures:

Professional microscope usage

Keen observation skills

Superior staining and bacterial growth

We are: The Pursuers

What is going on?

While other lab groups slack, ask questions, and talk...

Unsure!

Talking!

Focused.

...The Pursuers are hard at work, aware of the task before us.  We work efficiently, with precision and skill.

Intelligence.

Welcome to Medical Microbiology Lab. 2013.

The Matts

Murses

The Legend

3D Time

We are family.

Mission Impossible: Kill the Bacteria & Obligate Anaerobe Test Results...oh and DON'T pick that nose!!

Mission Impossible: Kill the Bacteria & Obligate Anaerobe Test Results...oh and DON'T pick that nose!!

Class: Tuesday November 12, 2013

Today we put our investigation into Mystery Bacteria J on hold to embark on a new and exciting mission: killing bacteria!! This little monsters are everywhere, so it's good to know how to nab 'em before they strike.  We decided to swab Matt's phone because, let's face it, it's probably pretty gross.  I mean we touch those things every day, carry them with us everywhere, set them down in God-knows-what, and even hold them up to our faces....see? Nasty!

 So, can we kill them? Let's find out! We worked with Jess and Mary Magree because there were 8 disinfectants/antibacterial cleaning agents to use, so we each tested four, using a swab from Matt's phone.  We tested...

2% Lysol spray (this is what we use to disinfect the counters in lab before and after class)

Lysol disinfectant spray (in Mandarin Orange) 

Foaming Antibacterial hand soap in Wildberry Freesia scent.  yum.

And Lysol disinfectant wipes.  This product as surfactants, quinines, as well as the disinfectant chemicals which rid the surface of organic material that would inhibit the ability of the disinfectant agents to kill the bacteria. 

Let us see if these products really do what they are telling us they do...

In the meantime, check THIS out!  Is it a light saber? Is it nuclear? What is it???

 That would be some UV radiation baby!!! UV light is supposedly very effective at killing all types of bacteria, including endospores.  So of course, we put it to the test by taking a small bit of each bacterial sample, mixing it into a nasty cocktail of bacterial floaters, and then zapped it with the SteriPen, which emanates concentrated UVB light waves.

Good work, Matt, you kill those bacteria! 

 We did a little bit of work today with Bacteria J, just to check the obligate anaerobe test.  This is the anoxalte chamber which, once it is sealed on, uses up all the oxygen so the bacteria have to grow in an anaerobic environment....















...or not.  Ours is not obligate anaerobe, which is supportive of our other findings thus far, that Bacteria J is a facultative anaerobe.  It needs at least a little bit of oxygen to grow, but is able to grow sparingly in low oxygen environment, as shown by the thioglycollate test.














Listen up people! Your mothers were right!! Don't pick your nose...wanna nose why? It's filled with Staphylococcus Aureus! Yikes!  But fear not, this particular strain is not necessarily dangerous.  Staph is actually all over the body, but in individualize with healthy immune systems it not colonize.  Still, if you are going to pick your nose, at the very least PLEASE don't eat it!

Swabbing Noses

Swabbing Noses

Class: Thursday, November 7

To continue with our practice as future nurses swabbing various parts of the body, we practiced swabbing noses today! 

We would grow the swab results in a blood culture and grow them, to check for Staphylococcus aureus infections. 

Blood Agar plate

Swabbing noses

Our results!

To test positive for a Staph infection, there would be lyses (holes) surround the bacteria growth into the blood agar, as well as a yellowish color of the bacteria.  Both students tested negative for Staph infections.

Fieldtrip to the Sewage Treatment Plant!

Fieldtrip to the Sewage Treatment Plant

Class: Tuesday, November 5

Today we learned visited the Sewage Treatment Plant! We were given a tour of the plant and we learned the important role that bacteria plays in the breakdown of waste.

First, we talked about how the water is treated.  The BOD (biochemical oxygen demand) levels are low because the water is dilute.  Why is it dilute?  This is because of underground water coming into the system as well as the high levels of rainfall we experience in the town.  Also, the population density is much lower than in earlier times when the treatment plant was built.   This is why the treatment plant switches right into secondary treatment of water and waste.

 Also to combat the influx of clean water into the waste treatment system, streams have been redirected into the river.

This is the anaerobic generator.  The purpose of this is to grow bacteria, to burn the sludge created, and then to use the charred sludge as fertilizer.  

Different class levels of pathogens exist.  It is possible to use thermophiles (bacteria that thrive in the heat) to grow the right class of sludge.

 We also discussed how phosphorous is becoming a precious element - that one idea is to begin extracting phosphorous during treatment.

 Peracetic acid is a disinfectant that is currently being used at this treatment center.  It is cheaper and more effective.  Not only that, but it is good for the water and the environment.  Many treatment centers that use other harsh chemicals need biochemical teams to come in and test the waters to makes sure that the levels are safe.  However, when peracetic acid reaches the river, it dissolves.

Your Dream Team: