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