Kevin Lance Lotharp

10/8/10

The Development of Antibiotic resistance in E. coli Summary

APA-format citation:
A, J. The Development of Antibiotic Resistance in E. coli. Retrieved from
http://share2.esd105.wednet.edu/mcmillend/02SciProj/jonatha/jonathana.html

Statement of reason for reading this book or selection:

This source was invaluable because it was generated by a student with a similar research problem to the one at hand.

Questions to be answered by this text:

1) How can the Kirby Bauer Method be applied to this type of project?
2) For how long is the culture of E. coli supposed to be incubated?
3) What is to be expected after this incubation period?
4) What is the basic set-up for any project of this sort?
5) How is the development of resistance determined?
6) What other materials not previously listed in the lab notebook for this project have been listed in the literature?
7) How can turbidity be determined and adjusted?

Answers to questions:
The Kirby Bauer Method is carried out in the same method that it is record in the lab notebook. The literature called for a colorimeter,
tests, Mueller Hinton agar plates, cotton swabs, normal saline at 0.9%, an incubator, calipers, BHI agar slants, E. coli, antibiotic
disks(in this case, ampicillin saturated disks), tryptic soy broth, antibiotic dilution(in this case, ampicillin dilution)1mg to 1mL, and
pipettes. First, take a sample of E. coli by dabbing the cotton swab on the provided culture. Smear this on the inside of one of the
given test tubes and add saline to the tube after doing so. Cap the tube and shake at a reasonable rate then place the tube in the
colorimeter. There were questions about turbidity and this is the solution to it. The calorimeter should be adjust so that the
transmission of the solution is 80% (which would compare to the turbidity of the 0.5 McFarland Standard). After creating this
solution, take another cotton swab and swab the E. coli solution all over the agar plate coated witht the MH agar. Place the antibiotic
disks on the agar plate equidistant from each other. Then the tryptic soy broth (TSB) must be configured. Add 2.1 ML of TSB, 200 mg
of ampicillin dilution, and then 200 mg of E. coli supspension to an empty test tube. Incubate the MH agar plate for a minimum of 24
hours or longer. Afterwards, measure the inhibition zones and record these diameters with calipers. For the following weeks of the
experiment, the bacterial suspensions were then made from the previous week’s TSB solutions.The development of resistance is
determined by the NCCLS interpretive standards.

Further questions not found in the text and/or things I am confused about:

1) What other antibiotics could be used in a manner similar to the way the ampicillin was used in the literature?
2) How long would this process take when conducted with more than one antibiotic?
3) Would a different culture of E. coli be needed for every different antibiotic or could more than one antibiotic be applied onto
the same plate?
4) For what is the BHI agar slant needed?
5) Where can TSB be found and how much would it cost?
6) What is the purpose of the saline solution?
7) How does one autoclave?
How this reading is related to my study:

This reading basically outlines the steps that are necessary to make the first part of the project feasible and a success. It provides both
help and instruction and also provides a new focus for the future of the project.

What don’t I understand from this source and/or what do I want to know more about after reading this
source?
Could the student that completed the experiment as described in the literature use his research for future problems and with other
applications?