Aseptic Technique and Bacterial Anatomy and Morphology

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Lab, Week

ASEPTIC TECHNIQUE AND BACTERIAL ANATOMY AND MORPHOLOGY

Introduction

Part I: Aseptic Technique

The purpose of this experiment is to become familiar with the specific microbiological technique known as the aseptic technique, which is used to avoid contaminating cultures.

In this case a pure culture of an unknown organism was introduced to a sterile medium of Phenol Red Glucose Broth Durham. The culture was obtained from a 52-year old male truck driver who is complaining to his doctor about pain in his lower abdominal and back. With a family history of prostate cancer, he worried that his sudden need to urinate during the night and infrequent appearance of blood in his urine might be bad signs. An ultrasound scan determined that the prostate gland was swollen. The physician sent a urine specimen to the lab for analysis. This organism was detected in large numbers in the urine.

After performing the aseptic techniques, it was found that if the proper techniques are not performed within a certain time frame the specimen will become contaminated.

Part II: Bacterial Anatomy

The purpose of this experiment is to become familiar and gain an understanding of how morphology and the arrangement of bacteria relate to staining techniques.

Sample 1

In this study a female patient who had a problem that had arisen suddenly after lunch, had allergic reaction to something that she ate or drank had sent her to the bathroom with cramps and copious diarrhea. She knew she was violently allergic to wheat gluten, but the waiter had assured her that the meal was wheat gluten-free. Her lawyer-husband had his investigators at the restaurant before they closed that night, testing for any evidence that would uncover the cause. This is one of the organisms that the investigators found in the dishes she had that day. A sample of the unknown organism was used to perform a gram stain procedure.

Sample 2

In this study a dead goat was found in the municipal swimming pool in the morning, the lifeguards were asked to remove it and clean the pool. Unfortunately, before they could add the chemicals, the city health department arrived to perform its weekly inspection. An organism was found in the water samples. This sample of the unknown organism was used to perform a gram stain procedure.

After performing the two bacterial anatomy studies, it was found that depending on the cell wall type, some cells will stain purple and others will stain pink. In gram staining if the results are gram positive cells, the cell walls are thick and will stain purple and if the results are gram negative. The cells can be found in different shapes and arrangements.

Procedure

Part I: Aseptic Technique

Open the VirtualUnknown Software.

Click the New button and select Aseptic Technique from the Select Exercise Topic or Subgroup dropdown box.

Enter Sample 1 Lab #5 in the Enter Unknown Identifier field.

Do not click the box allowing autoinoculation.

Observe the Gram reaction of the unknown and indicate the results of the gram stain by selecting the result that pertains to the observation and record the result by clicking record gram reaction.

All of the following will be needed for the experiments: watch or clock with a second hand, test tubes, Bunsen burner, inoculation tools, and an incubator.

From the media dropdown menu, select a Phenol Red Glucose Broth Durham Tube and label it prgb. The tube will then appear on the desktop along with a tube of the unknown organism, the sterile phenol red glucose broth on the right and the unknown of the left.

In order to determine the amount of time it would take for contamination to occur, you will need to have the watch or clock handy. Begin by removing the cap from the test tubes and start timing. You will need to observe the Inoculation/Contamination traffic lights, when the contamination traffic light turns red you will stop timing and recap...
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