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.