Research Proposal

Susceptibility of Antibacterial Chemicals on

Pseudomonas aeruginosa  

 

Researched by: Clayton Brady

Biology Department of Tennessee Technological University

Cookeville, TN 38501

Keywords

Pseudomonas aeruginosa

Antibacterial agents/chemicals

Bacteria

Susceptibility

Resistance

Table of Contents

  1. Title

  2. Keywords

  3. Table of Contents

  4. Project Summary

  5. Introduction

  6. Methods and Materials

  7. Expected Results and Benefits

  8. Project Timeline 

  9. Literature Cited

  10. Budget

Project Summary

    Antimicrobial agents are used in in our homes as well as hospitals to kill or hinder dangerous bacteria.  Bacteria can be resistant to many of these agents.  Pseudomonas aeruginosa will be used as a test subject for this experiment for the sole reason of its resistance to many different agents.  This subject is difficult to isolate and will prove to be a more than formidable foe for these antibacterial agents.  The objective of my research is to determine if the antibacterial sprays we use so abundantly today can in fact render potentially hazardous bacteria, such as Pseudomonas aeruginosa, non-viable.

Introduction

        Many diseases arise from bacteria that grow resistant to the agents that are made to combat them.  New research is consistently being done combat these bacterium.  Through new research, scientists can develop new antimicrobial agents and antibiotics to ensure the elimination of a bacterium for a limited period of time.  Much research has been done on Pseudomonas aeruginosa.  Pseudomonas  aeruginosa is responsible for 16% of nosocomial pneumonia cases , 12% of hospital-acquired urinary tract infections , 8% of surgical wound infections , and 10% of bloodstream infections. (Delden & Iglewski, Volume 4, Number 4)

          It has been proven that Pseudomonas aeruginosa develops plasmids that are that are able to transfer their genes through the processes of transduction and conjugation.  Transduction is the process of transferring genetic information from cell to cell through the mediation of a virus (phage) particle.  Conjugation is a unidirectional transfer of genetic material between cells by cell-to-cell contact.   By transferring their genetic material from cell to cell by this means, Pseudomonas aeruginosa is able to elude antimicrobial agents and antibiotics and survive. Thus making it a perfect "super subject" to test chemicals that claim to kill harmful bacteria.

         I will disprove the Null hypothesis by proving that Pseudomonas aeruginosa is susceptible to certain antibacterial chemicals.  Strains of Pseudomonas aeruginosa from a synthetic stock will be tested against individual tests commonly used in microbiology laboratories, as well as chemicals commonly used to disinfect out homes and hospitals.  All mediums will be collected and examined to determine if Pseudomonas aeruginosa is susceptible to this test.  The objective of my research is to determine if the antibacterial sprays we use so abundantly today can in fact render potentially hazardous bacteria, such as Pseudomonas aeruginosa, non-viable.  By doing this I hope to find how safe we are using chemicals and sprays that we trust in our homes to keep us healthy.

Methods and Materials

 The sample used in this experiment came from a synthetic stock.

      Test includes the following:

 Mueller Hinton / Pseudomonas Agar Disc Diffusion Assay Tests (This technique allows you to          test discs on top of the agar to see if they are   resistant or susceptible.)   (Goss 62-66)

Other materials that are needed during the experiment include:

  Using aseptic techniques, the experiment began with taking an inoculating loop, sterilizing it by way of heating it, and obtaining a sample of the bacteria being used.  (It is important to sterilize the loop in order to prevent any type of contamination of the bacteria from outside sources.)  After heating the loop and allowing the loop to cool, the bacterium was then streaked onto an agar surface inside a Petri dish.  As soon as this process was finished, the loop was sterilized again to kill off any excess bacteria that was left on the loop.  The bacteria was then exposed to discs dipped in each of the three chemicals.  The dishes were separated into quadrants for each of the three chemicals being used with one quadrant being used as a control with only a sterile disc dropped on the agar.

Expected Results and Benefits

I expect that, after careful aseptic experimentation, I will be able to kill Pseudomonas aeruginosa with a the antibacterial chemicals that will be used in the experimental process.

Benefits of this experiment will include a better understanding of this bacteria as well as knowledge of bacterial protection for home and workplace use.

Project Timeline

 The entire experimentation will take approximately 3 days to complete.  

1st Day:  Preparation of all agar plates and apparatuses.  Streaking of plates.  Incubation.

2nd Day:  This day will be used to allow the bacteria to grow.

3rd Day:  Determination of susceptibility and resistance of the bacteria to individual chemicals as well as pictures of the apparatuses.

 

Literature Cited

Delden, C.V. , Iglewski, B.H.  1998.  Cell-to-Cell Signaling and Pseudomonas aeruginosa Infections.  Emerging Infectious Diseases Volume 4, Number 4.  www.cdc.gov/ncidod/eid/vol4no4/vandelden.htm

Goss, S.  2001-2002. Control of Microbial Growth.  Health Science Microbiology Laboratory Manual.  62-66

Budget

          I am currently working to have the entire project funded by Dr. Susan Goss of the Tennessee Tech Biology Department.

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