Streak a Bacterial Culture Anticeptics

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Introduction
The working hypothesis being tested is if any of the three products; Scope Mouthwash (Scope), LISTERINE® Antiseptic Mouthwash (Listerine), Bactine Original First Aid Liquid (Bactine), and Povidone-Iodine Solution, 10% Topical Microbicide Antiseptic (Iodine) are an appropriate antiseptic to can kill a sample bacterial contaminant found in the local environment, and if so, which is the most effective? The purpose of this experiment is to determine the antibacterial effectiveness of Scope, Listerine, Bactine, and Iodine in relation to each other. Scope, Listerine, Bactine, and Iodine each are believed to be effective antiseptics: * Scope Mouthwash - Kills 99% of bad breath germs (in lab tests) http://www.scopemouthwash.com/scope-products/scope-mouthwash-original-mint.aspx * LISTERINE® Antiseptic Mouthwash - This effective rinse kills germs in virtually 100% of the mouth, penetrating to kill plaque biofilm. http://www.jjdentalprofessional.com/deepestclean * Bactine Original First Aid Liquid has provided soothing infection protection for over 50 years. http://www.bactine.com/original.htm * Iodine - A commonly used antimicrobial agent is Povidone-iodine (Betadine), a complex of iodine, the bactericidal component, with polyvinylpyrrolidone (Povidone), a synthetic polymer. http://www.ncbi.nlm.nih.gov/pubmed/9474112

Materials and Methods
1) Step 1 – Collecting and growing a sample bacterial contaminant found in the local environment. a) Materials:
i) Sterile cotton swabs.
ii) Tube of sterile distilled water.
iii) Petri dishes with nutrient agar
iv) China Marker
v) Object to be tested (all are from the front of the class room): (1) Teachers Chair
(2) Computer Keyboard
(3) Computer Mouse
(4) Teachers Desktop
b) Method:
vi) draw, with a china marker, on the outside of the agar portion, not the lid, of a Petri dish divided it into 4 sections labeled 1-4 to indicate where sample materials were taken from. vii) Use a sterile cotton swab moistened in sterile water rotate the side of the swap across the object to be tested. viii) Transfer the contaminants by smearing and rotating the swap over the surface of the agar on the Petri dish in the quadrant corresponding to the assigned sample number. ix) The inoculate dish was Incubated upside down for more than a week to produce a healthy growth. 1) Step 2 – Gram Staining

x) Materials:
(5) Microscope slide
(6) Gram stain reagents
(a) crystal violet
(b) Gram's iodine
(c) 95% ethyl alcohol
(d) Safranin
(7) Sample of bacterial contaminant found in the local environment (Culture from Step 1) c) Method:
xi) Label the slides
xii) A droplet of distilled water is placed on the slide xiii) Bacteria is obtained from the agar using a sterile loop and mixed with the water on the slide xiv) Air dry and heat fix

xv) Cover with Crystal Violet (Primary Stain) for 20 seconds. xvi) Gently rinse off the stain with water and shake off the excess xvii) Cover with Gram's Iodine for one minute. (Mordant) xviii) Pour off the Gram's iodine.

xix) Run 95% Ethyl Alcohol (Decolorizing Agent) down the slide until the solvent runs clear, about 10-20 seconds xx) Rinse with water to stop the action of the alcohol xxi) Cover with Safranin (Counter Stain) for 20 seconds xxii) Gently rinse off the stain with water

xxiii) Blot with paper towel
2) Step 3 – Antiseptics Plate Streak Method for Colony Isolation 1- draw, with a china marker, on the outside of the agar portion, not the lid, of a Petri dish divided it into 4 sections labeled 1-4 to the areas that will be tested with antiseptic. 2-...
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