Lab 6 – Selective and Differential Media Ninoska Garcia-Ortiz 063 053 2 Objective: • • To learn the difference between

selective and differential media. To learn how to use both types of media

Theory: Selective media – one or several chemical inhibiting growth of most types of microorganisms, exception select few Differential media – recognition (differentiation) between 2 or more types of microorganisms. Some media can be both selective and differential. Four types of agar: Mannitol Salts Agar (MSA) – used for Staphylococcus. Differentiates between S. aureus and S. Epidermis. High concentration (7.5%) of sodium chloride selects for micrococci staphylococci. Mannitol acidifiers (eg. S. aureus) have yellow zones around their colonies due to change of the phenol red indicator by the acid. Macconkey Agar (MA) – used for Gram negative enteric bacteria. Differentiates between lactose fermenters and nonfermenters. Gram – enteric bacilli grow. Gram + bacteria growth inhibited by crystal violet. Differentiates between brick red lactose fermenting (lac+) and transparent lactose nonfermenting (lac-) bacilli. Pathogenic enteric bacteria tend to be lac- bacilli (eg. Shigella and Salmonella) Tellurite Glycine Agar (TGA) – used for coagulase-positive staphylococci. Will grow as black colonies. Coagulase-negative and other microorganisms are inhibited. Blood Agar (BA) – used to grow nutritionally demanding organisms. Differentiates between those that lyse red blood and those that do not. Alpha-hemolytic organisms produce green, olive zone around colonies due to oxidative effect of peroxide waste on heme. Beta-hemolytic organisms produce clear zone around their colonies, due to lysis of red blood cells by bacterial exoenzymes known as hemolysins. Gamma-hemolytic organisms will exhibit no detectable change around colonies.

Questions: 1– Mannitol Salts Agar – selective media. • Specifically selects for micrococci and staphylococci by way of the nutrients that it contains. Macconkey Agar - both selective and differential. • Selective due to the fact that it allows Gram negative enteric bacilli to grow and inhibits the growth of Gram positive bacteria. • Differential because it allows for differentiation of lactose fermenting and non lactose fermenting Gram negative enteric bacilli such as E.coli and P.vulgaris Tellurite Glycine Agar - selective media. • specifically tests for coagulase-positive staphylococci (which will grow after 24hrs.) Coagulase-negative staphylococci and other organisms are inhibited. Blood Agar – differential media. • Enriched medium which will allow for the growth of most micro-organisms. • Serves to differentiate between alpha-hemolytic organisms that produce a green olive zone, beta-hemolytic organisms that will produce a clear zone and gamma haemolytic organisms that will exhibit no detectable change. 2– Alpha hemolysis - caused by oxidative effect of peroxide waste on heme Beta-hemolysis – caused by lysis of the red blood cells by the bacterial exoenzymes known as hemolysis. 3– Macconkey Agar, which is both a selective and a differential media, could be used to determine the presence of E. coli. This medium allows for the differentiation of lactose fermenting and non lactose fermenting Gram negative enteric bacilli such as E.coli and P.vulgaris. 4– The results for all the plates were as expected, except the growth on the Tellurite Glycerine Agar plates. The plate with the S. eareus gave the expected results, however; the plate with the S. epidermis had similar results. There was not supposed to be any growth in this plate. The growth can perhaps be explained by the fact that the plate was not inspected in the recommended 24 hour time frame, but instead 1 week later. This theory is further enforced by the evident “slower” growth on the plate that should not have had any growth.

Conclusion: The objective of the lab, • • To learn the difference between selective and differential media, and To learn how to use both types of media,

was achieved. The use of four different types of Agar were used to attain the objective. In the case of the Tellurite Glycerine Agar, we saw the importance of analyzing results of time sensitive experiments. The assumption that the growth was due to prolonged exposure is based on the fact that we were already told that coagulase-positive staphylococci would grow black colonies within 24 hours. The fact that there was growth in both plates, is an indication that the experiment was not done correctly. In this case, we are able to identify the infraction, analysis one week later, instead of 24 hours later.

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