Many Thanks!!

The past few weeks have been a bit stressful as the semester is coming to a rapid close. My partner and I have officially been able to complete our slides for our presentation. What a wonderful feeling it is. I want to also give many thanks to our fellow S.T.E.M. mates, as they helped us a lot by giving us necessary feedback on our presentation. I welcome the constructive criticism. I am very grateful to have the ability to work with such an awesome group of people. I cannot wait to see the results of everyone's hard work this semester.

Adrenaline Rush

Yikes! Nothing like an adrenaline rush when you forget to utilize a test that’s crucial for your project. I am so glad we had a fellow STEM member to guide us in the right direction. I’m also glad that we kept our culture plates. If we didn’t I’m not entirely sure what would happen. We probably would've had an extremely hard time identifying the bacteria. We ended up scrambling to get the results for FTM. This test is important because it will tell us whether or not our bacteria is aerobic, anaerobic, or facultative.

Name that Bacteria

As this semester is rapidly coming to a close so is our research project. We have officially finished all of the tests that needed to be completed. The only thing left for us to do is take that information and start identifying our bacteria. I'm realizing that this may take awhile as we have about 30 or more specimens. Next week we will start to talk about our plans for our poster. I've never done one of these before so it will be interesting. Wish us luck!

Dreamy Draw

This has been a very busy week in the lab. I went hiking at Dreamy Draw on Sunday to gather more samples for testing. I have officially isolated 24 colonies this week. Meaning next week is going to be even busier as we will have to perform various tests on all of the individual colonies. I decided I would start to share some interesting facts about Creosote as I learn myself. Creosote bushes form symbiotic relationships (mutualism) with bacteria, fungi and algae. These microbial communities allow the Creosote to gather more phosphorous and nitrogen during rainfall. Making this desert plant more advanced than other desert plants.

The black parts of the Creosote are where the microbial communities thrive. (Bacteria, algae, fungi)

Dreamy Draw

Positivity!

Today while I was checking the tests we performed the other day, we finally had a positive Simmons Citrate test. This test can be used to determine certain coliforms (e.g. Enterobacter aerogenes). This bacteria can naturally occur in soil. This test helps us find out if a bacteria uses sodium citrate as it's sole carbon source. We also may have identified one bacteria so far for another one of our samples. It's called deinococcus radiodurans. This bacteria can survive in the most extreme conditions (cold, dehydration, etc.). It made The Guinness Book of World Records for the toughest bacteria.

Right Track

I finally figured out a system for testing the bacteria. After a few weeks of trial and error I am on the right track. Considering that I have so many tests to perform for one bacteria I changed my hours throughout the week to ensure consistency. This will also help me with prepping for my work the following week. Time flies when you're extremely busy. I can hardly believe that this semester is already halfway over. That being said I have some work to catch up on. I look forward to smoother weeks ahead.

Moving On

After a rocky start last week we are finally getting back on track. We had so much testing to catch up on. Some of these tests I’ve never performed before so it was awesome to see them in action. For instance, the catalase test, we took slides and placed our bacteria on them then put hydrogen peroxide on top of the bacteria. Only 3 of the bacteria possessed the catalase enzyme (positive). I’m looking forward to next week. We are going to learn how to make our own TSA plates and a few other things we will need for our testing.

Next weeks bacteria we have to isolate

Next weeks bacteria we have to isolate

Bumps in the Road

This week has not gone as planned. I have had a few setbacks with my project. Last week Mario and I were finally getting things together for our project which required a lot of research. So this week I finally thought I had everything down as far as the testing methods and such. When we came in this Tuesday to start making gram stains and testing the bacteria, we got a notice that the school was closing at noon due to the thunderstorms. The back of my house also got flooded the same day. Talk about adventures. I decided to not stress myself out and plan on starting anew next week. On the bright side,  I think we have finally found the perfect method of removing bacteria from the phyllosphere. 

Tween (used to remove peripheral proteins)

Creosote Week #1

I'm officially moving on to my new project. I will be working on the Creosote project with Mario. So far the only thing we have done is a lot of reading and research. We are trying to figure out what is the best method for extracting the bacteria from Creosote leaves. One approach we will be working with is smearing the leaves directly onto the agar plate. The second procedure is the one we’ve been researching in depth. It is a solution called Tween 20, which is a detergent that will substantially help remove the bacteria from the leaves without disrupting (cell lysis) the plant. This coming week we will be testing to see which is the best method. Wish us luck!

Creosote Bush

Unknown Bacteria Week 3

Week 3: Unknown Bacteria

The last phase of testing on my unknown bacteria is officially complete.. The gelatin was placed in the incubator at 37℃ for five days. After this period I then placed the gelatin in the refrigerator to cool it for 6 hours. The gelatin never solidified. Meaning, the organism was able to break it down. Through the gelatin testing I found the bacteria to be positive for hydrolysis. So through these various methods of testing, the bacteria finally revealed itself, Serratia marcescens. This organism is known to cause common illnesses including respiratory infections and urinary tract infections.

Serratia marcescens

Unknown Bacteria Week 2

Week 2: Unknown Bacteria S8772274699M

Materials and Methods

The next step was to perform the oxidase test. Initially, the bacteria sample was inconclusive. The oxidase check was fulfilled by using the pure culture plate along with using the broth. The same process was conducted on 9/11/18 to retest, and the results came back oxidase negative. I proceeded to the sugar fermentation test. By using a loop to obtain a small number of bacteria and added it to lactose and glucose. These were then placed in the incubator at 37.9℃ for five days. Results showed that the bacteria was negative for carbohydrate fermentation (lactose) but showed positive for gas and fermentation (glucose). The first phase of the SIM test was carried out. The isolated bacterial colonies were positioned into the SIM medium.  Then placed into the incubator set at 37.3℃ for 24 hours. The SIM test showed positive for motility. I will be continuing to the next phase of the project which is gelatin testing. Those results will be known next week and should tell me what type of unknown bacteria S8772274699M is.

Oxidase Test	Sugar Fermentation	SIM Test
Swab Oxidase	Lactose Glucose Durham tube	SIM Needle

Lactose Fermentation

Glucose Fermentation

SIM Test

Unknown Bacteria Week 1

Introduction

The purpose of this lab was to be able to establish proper TSA culture techniques. Learn how to Gram stain slides. Along with being able to identify the unknown bacteria using methods of testing, evaluation, and process of elimination.

Materials and methods

The instructor provided a test tube labeled S8772274699M for testing of the unknown bacteria. The first procedure was to isolate the bacteria and gain a pure culture.  Obtained by using a Trypticase Soy Agar (TSA) plate. Two culture plates were produced to isolate bacterial colonies. The first streak was in a “T” method the other was done using the Lawn technique. Leaving the TSA plates for five days at room temperature allowed the bacterial colonies to grow. Upon retrieving the bacteria, the colony morphology observations were as follows: punctiform, margin entire, elevation flat, surface smooth, translucent, and red coloring. The next step was making gram staining glass slides to be able to view bacteria under a microscope. The unknown bacteria were Gram-negative, as the organisms under the microscope were pink. Continuation of this experiment to be conducted next week.  

Materials

Plates	Smear Preparation	Gram Staining
Trypticase Soy Agar (TSA) plates Inoculating loop S8772274699M Broth	Glass slide Distilled water Inoculating needle Pure bacteria culture	Crystal violet Gram iodine Alcohol decolorizer Safranin Distilled water Bibulous paper