Differential & Selective Media in Microbiology

Every microbiologist eventually has to grow cultures of bacteria for their experiments. In this lesson, we will
investigate selective and di erential media, two techniques to make culturing bacterial species a little more
e cient.

Selective and Differential Foods?

I bet that every one of us knows someone who is allergic to peanuts. Standing in a crowded room,
it would be impossible to identify the individuals with this allergy simply by looking at them -
assuming no one is wearing a medical alert bracelet. But what if we really wanted to know who was
allergic to peanuts and who was not? What could we do?

One option, and I'm not suggesting you try this at home, would be to give everyone in the room a
cookie laced with peanuts. Very quickly, the allergic people in the room would start showing
symptoms. They might start having trouble breathing, swell up or start vomiting. In any case, it
would be very easy to di erentiate the mixed population into allergic and non-allergic groups. The
peanuts would serve as a di erential food.

Let's take this a little further. Now that you know who is allergic to peanuts, suppose you want to
encourage the survival of peanut-resistant people while inhibiting the survival of the peanut-
allergic. All you would have to do is provide these people only peanut-based foods. People that
could eat peanuts would be ne. People that could not would either starve or die of an allergic
reaction. You have just selected for the peanut-resistant people. Here, the peanuts would serve as
a selective food.

Culture Media
These experiments might seem a bit cruel and out of place in the human world, but they are
occurring in the bacterial world daily in microbiology labs, hospitals and classrooms. Learning how
to grow and manipulate bacteria is a very useful skill for a microbiologist. Let's quickly review a few
key concepts before we get to the meat of this lesson. Bacterial culturing is the process of
growing bacteria in a medium. A culture can contain only one species of bacteria, called a pure
culture, or can contain many di erent species, called a mixed culture.

A medium is a liquid or gel designed to support the growth of a bacterial culture. The medium has
to contain everything that the bacteria needs for growth and survival. There are countless media
variations depending on what bacterial species you are attempting to grow. And, just like the
peanuts in the introduction, you can even include ingredients to di erentiate one species from
another or select for the growth of a certain type of bacteria while inhibiting others.

Differential Media
The rst type of media we'll investigate is di erential media. A di erential media contains speci c
ingredients or chemicals that help us to visually distinguish which species do or don't carry out a
speci c biochemical process. To clarify this de nition, let's look at an example of a di erential
medium.

Blood agar is a di erential medium that distinguishes bacterial species by their ability to break
down the red blood cells included in the media. Blood agar is often used to distinguish between
the di erent species of pathogenic Streptococcus bacteria. The di erent types of Strep each have a
predictable pattern of hemolysis, which is simply the breakdown of red blood cells. Alpha-
hemolytic Streptococcus, like those that cause pneumonia, produce a narrow band of slimy
discoloration around the colony during the partial breakdown of the red blood cells.

Beta-hemolytic Streptococcus, like those that cause potentially deadly infections in newborns,
produce a completely clear zone around the colony during the complete breakdown of the red
blood cells. Gamma-hemolytic Enterococcus, which used to be called Group D Streptococcus, don't
change the media appearance at all since they can't break down the red blood cells. In these
pictures, you can see how simply growing these bacteria on blood agar can help determine what
type of Streptococcus you're dealing with.

Selective Media
The next type of media is selective media. A selective media is composed of speci c ingredients to
inhibit the growth of certain species of microbes in a mixed culture while allowing others to grow.
Again, in order to fully grasp this de nition, let's look at an example of a selective medium.

MacConkey agar is a selective medium containing bile salts and crystal violet that inhibits the
growth of Gram-positive bacteria while allowing for the growth of Gram-negative bacteria.
MacConkey agar is often used when investigating and culturing fecal bacteria. Many disease-
causing bacteria found in human feces are Gram-negative, like E. coli, a common cause of food
poisoning. Gram-positive species are unable to grow in the presence of bile salts and crystal violet,
ensuring that more colonies will be the Gram-negative pathogens of interest.

Selective and Differential Media

At this point, you should have a good handle on why microbiologists would use selective and
di erential media. But in the intro scenario, we used peanuts as both a di erential and a selective
agent. Today, many media recipes have been developed to be both selective and di erential.
Ingredients have been included that inhibit the growth of certain bacterial species while also
distinguishing between the di erent species that are able to grow. The combination eliminates the
need for culturing twice: once to select for a speci c group and the second time to di erentiate
between members of that group.

The MacConkey agar we talked about as an example of a selective media is also a di erential
media. In addition to the selective bile salts and crystal violet, MacConkey agar contains a pH
indicator and the sugar lactose. Some Gram-negative bacteria are able to ferment lactose,
producing an acidic byproduct. The pH indicator makes these colonies appear red. Non-lactose
fermenters will be colorless. So, the MacConkey agar selects for Gram-negative bacteria and
di erentiates between lactose fermenters and non-fermenters.

There are countless examples of selective and di erential media. The two presented here are just
the tip of the iceberg. Some select by including very speci c energy sources or antibiotics in the
media. Others are able to di erentiate through gas production, metabolic waste production or the
presence of genetically modi ed DNA. The possibilities are endless.

Lesson Summary
Let's review. In order to study bacteria, microbiologists need to grow the cells in the lab; this is
called culturing. Sometimes a bacterial sample is mixed, meaning it contains multiple species of
bacteria, and sometimes only one or a few species are of interest. In order to only culture the
desired bacteria, a scientist can change the components of the media used to grow the bacteria.

A selective media is composed of speci c ingredients to inhibit the growth of certain species of
microbes in a mixed culture while allowing others to grow. MacConkey agar is a selective medium
containing bile salts and crystal violet to inhibit the growth of Gram-positive bacteria while allowing
for the growth of Gram-negative species.

A di erential media contains speci c ingredients or chemicals that allow the observer to visually
distinguish which species possess and which species lack a speci c biochemical process. Blood
agar is a di erential medium that distinguishes bacterial species by their ability to break down red
blood cells.

The ability to break down the cells will cause a change in the color of the blood agar. Some media
are both di erential and selective. Combining the two can often eliminate having to perform
multiple cultures, saving time and money for the microbiologist. MacConkey agar can both select
for Gram-negative bacteria and di erentiate lactose fermenting and non-fermenting bacteria.

Learning Outcomes
When this lesson is done, you should be able to:

De ne what di erential and selective media are

Di erentiate between MacConkey agar and blood agar