General Microbiology Lab Report

Name: Loan Do.

Date:

03/14/2012

Title of Exercise: Selective and Differential Media Question: Is my unknown #590 gram positive? Experimental rationale: Selective media can completely inhibit and/or slowly down the growth of certain species of bacteria. There are some species of bacteria that are unaffected by the toxic compounds that are contained within the selective media and thus can grown within the environment without difficulty. For example, Eosin Methylene Blue Agar (EMB) contains two dyes that inhibit the growth of gram positive bacteria, however, allow the growth of gram negative bacteria. EMB is said to be selective for growth of gram negative bacteria. The color of growth of the bacteria within the media can be identified as dark pink or purple in color. Phenylethyl Alcohol Agar (PEA) is a media that tests for the growth of gram positive bacteria. The media contains alcohol which aids in the denature of the outer membrane of gram negative cell walls, thus allowing gram positive bacteria to grow and inhibiting the growth of gram negative bacteria. The color of the media is yellow/tan in color. EMB is also a differential media in the sense that it contains chemical compounds that affects the appearance of the colony of the bacteria of different species. If a bacteria ferments lactose using EMB media, it will produce either dark purple colonies or a metallic green sheen.[1] A summary of the procedure is: 1) 2) 3) 4) Divide 2 plates of EMB and 1 plate of PEA into quadrants. Decide on one positive control and one negative control for each side of one EMB and one PEA media. Make a 2 cm streak on the plate on the EMB and PEA. Incubate plates for 48 hours at 35 degrees Celsius.

Controls: My unknown #590; negative control for the PEA selective media was Escherichia coli because E. coli is gram negative and therefore will not grow on PEA selective media, which selects for growth of gram positive bacteria; and positive control for the PEA selective media was

Enterococcus faecalis because this bacteria is gram positive and PEA selects for growth of gram positive bacteria.

Expected Results: If my unknown bacteria is gram positive, then there will be growth on the PEA media and no there will be growth on the EMB media. Actual Results: My unknown #590 showed growth on the PEA media and showed no growth on the EMB media.

EMB Selective Conclusion:

PEA Selective

Based on the results, my unknown bacteria #590 is gram positive. According to the EMB media above, which selects for growth of gram negative bacteria, E. coli can grow in this media, as evidenced by my experiment. The lack of growth of my unknown #590 on my EMB plate is a negative result and I compared this to my positive control. This revealed substantial growth difference between the two controls. Additionally, PEA selects for gram positive bacteria and my unknown #590 grew on PEA. This is a positive result and I compared this to my negative control, E. coli, which did not grow on this media. Therefore, based on my experiment, it can be concluded that my unknown #590 is gram positive. Next step: To identify my unknowns oxygen and temperature requirements for optimal bacterial growth, I will perform a physical requirements exercise for growth (oxygen requirements) using BHI molten agar shakes, TSA plates and unknown cultures (in broth) along with TSB tubes

(temperature requirements).2 References: Gutierrez P. Microbiology Exercises. Coastline Community College; 2012. 67 p.
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Dichotomous key