MacConkey Agar

MacConkey agar is both selective and differential. It contains bile salts and the dye
crystal violet, which inhibit the growth of gram-positive bacteria and select for
gram-negative bacteria. It also contains the carbohydrate lactose, which allows
differentiation of gram-negative bacteria based on their ability to ferment lactose.
Organisms which ferment lactose produce acid end-products which react with the pH
indicator neutral red, and produce a pink color.

Quadrant 1: Growth on the plate indicates the organism, Enterobacter aerogenes, is

not inhibited by bile salts and crystal violet and is a gram-negative bacterium. The
pink color of the bacterial growth indicates E. aerogenes is able to ferment lactose.

Quadrant 2: Growth on the plate indicates the organism, Escherichia coli, is not
inhibited by bile salts and crystal violet and is a gram-negative bacterium. The pink
color of the bacterial growth indicates E. coli is able to ferment lactose.

Quadrant 3: Absence of growth indicates the organism, Staphylococcus epidermidis,

is inhibited by bile salts and crystal violet and is a gram-positive bacterium.

Quadrant 4: Growth on the plate indicates the organism, Salmonella typhimurium, is

not inhibited by bile salts and crystal violet and is a gram-negative bacterium. The
absence of color in the bacterial growth indicates S. typhimurium is unable to ferment
lactose.
Back to Tutorial

File:MacConkey agar with LF and LF

colonies.jpg
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Summary[edit]
Description English: MacConkey's agar showing both lactose and non-lactose fermenting
 colonies. Lactose fermenting colonies are pink whereas non-lactose fermenting ones
are colourless or appear same as the medium.
Date 28 June 2007
Source Own work
Author Medimicro

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{{Information |
Description={{en|
1=MacConkey's agar showing
both lactose and non-lactose
2,304 × Medimicr fermenting colonies. Lactose
curren 16:21, 6 August
1,728 (757 o (talk | fermenting colonies are pink
t 2008
KB) contribs) whereas non-lactose
fermenting ones are colourless
or appear same as the
medium.}} |Source=Own work
by upload
{{Information |
Description={{en|
1=MacConkey's agar showing
both lactose and non-lactose
2,304 × Medimicr fermenting colonies. Lactose
16:13, 6 August
1,728 (757 o (talk | fermenting colonies are pink
2008
KB) contribs) whereas non-lactose
fermenting ones are colourless
or appear same as the
medium.}} |Source=Own work
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Cultivation Media for Bacteria
Isolation of bacteria is accomplished by growing ("culturing") them on the surface of solid
nutrient media. Such a medium normally consists of a mixture of protein digests (peptone,
tryptone) and inorganic salts, hardened by the addition of 1.5% agar. Examples of standard
general purpose media that will support the growth of a wide variety of bacteria include nutrient
agar, tryptic soy agar, and brain heart infusion agar. A medium may be enriched, by the addition
of blood or serum. Examples of enriched media include sheep blood agar and chocolate (heated
blood) agar.

Selective media contain ingredients that inhibit the growth of some organisms but allow others to
grow. For example, mannitol salt agar contains a high concentration of sodium chloride that
inhibits the growth of most organisms but permits staphylococci to grow.

Differential media contain compounds that allow groups of microorganisms to be visually

distinguished by the appearance of the colony or the surrounding media, usually on the basis of
some biochemical difference between the two groups. Blood agar is one type of differential
medium, allowing bacteria to be distinguished by the type of hemolysis produced. Some
differential media are also selective, for example, standard enteric agars such as MacConkey and
EMB agars, which are selective for gram-negative coliforms and can differentiate lactose-
fermenting and non-lactose-fermenting bacteria.
Several examples of commonly used bacteriological media, as well as examples with one or
more types of bacteria cultured on them are shown below. Carefully examine the plates and
observe the colony morphology, colors, and patterns of growth (or no growth) that occurs. This
information can be valuable in the preliminary identification of pathogens in case studies.

Common Bacteriologic Media

Tryptic Soy Agar (TSA)

Tryptic Soy Agar - uninoculated

Type: General

Purpose: Cultivation of non-fastidious bacteria

Interpretation: Growth indicates non-fastidious bacteria present

Examples

Chocolate Agar

Chocolate Agar - uninoculated

Type: Enriched

Purpose: Cultivation of fastidious organisms such as Neisseria or Haemophilus sp.

Interpretation: Some organisms grow on Chocolate that do not grow on standard media

Examples

Thayer-Martin Agar
Thayer-Martin Agar - uninoculated

Type: Enriched and selective; contains antibiotics colistin (kills gram-negative coliforms), vancomycin
(kills gram-positives), nystatin (kills fungi)

Purpose: To select for fastidious organisms, such as N. gonorrhoeae, in patient samples containing large
numbers of normal flora, such as in the female genital tract

Examples

MacConkey (lactose) Agar

MacConkey Agar - uninoculated

Type: Selective and differential

Purpose: Contains bile salts and crystal violet which selects for gram-negative enterics, differentiates
lactose-fermenters from non-fermenters. Can include sugars other than lactose for further
differentiation (for example, to differentiate enterohemorrhagic E. coli (EHEC), which does not ferment
sorbitol, from other E. coli types which do.)

Interpretation: Selects for non-fastidious gram-negatives; red colonies indicate fermentation of lactose,
white indicates no fermentation of lactose

Examples

Eosin-methylene Blue Agar (EMB)

Eosin Methylene Blue Agar - uninoculated

Type: Differential (lactose) and selective (dye inhibition and precipitation at acid pH)

Purpose: Differentiates lactose fermenters (E. coli) from non-fermenters (Salmonella, Shigella)

Interpretation: Lactose fermenters blue/black; non-fermenters colorless or light purple

Examples

Hektoen Agar

Hektoen - uninoculated

Type: Selective and differential

Purpose: Detects lactose fermentation, H2S production, inhibits non-enterics

Interpretation: Lactose fermenters yellow or salmon, non-fermenters colorless; H 2S production produces

black precipitate

Examples

Mannitol Salt Agar

Mannitol Salt Agar - uninoculated

Type: Selective and differential

Purpose: Selects for Staphylococci, which grow at high salt concentrations; differentiates
Staphylococcus aureus from other Staphylococci

Interpretation: Staphylococcus aureus is yellow (ferments mannitol), other staphylococci are white

Examples

Triple Sugar Iron Agar (TSI)

Triple Sugar Iron Agar - uninoculated

Type: Multi-purpose, differential

Purpose: Detects glucose, lactose, sucrose fermentation; gas and H2S production. (E. coli → A/AG;
Salmonella → K/A+G; Shigella → K/A; Ps. aeruginosa → K/K)

Interpretation: Yellow butt, red slant (K/A) = ferments glucose only; yellow butt and slant (A/A) =
ferments glucose + lactose and/or sucrose; red but and slant (K/K) = non-fermenter of all 3 sugars; black
(+) = H2S; bubbles (G) = gas production

Examples

Hemolytic Reactions Observed on Blood Agar

Observation of the hemolytic reactions on sheep blood agar is a very useful tool in the
preliminary identification of bacteria, particularly streptococci. The types of hemolysis are
defined as follows:

alpha (α) hemolysis: An indistinct zone of partial destruction of red blood cells (RBCs) appears
around the colony, often accompanied by a greenish to brownish discoloration of the medium.
Streptococcus pneumoniae and many oral streptococci are α hemolytic.

beta (β) hemolysis: A clear, colorless zone appears around the colonies, in which the RBCs have
undergone complete lysis. Streptococcus pyogenes, S. agalactiae, and several other species of
streptococci are β hemolytic. Many other bacteria besides streptococci can be β hemolytic,
including Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogenes, etc., and
hemolytic reactions can also be a useful diagnostic tool for these organisms.
no (γ) hemolysis: No apparent hemolytic activity or discoloration is produced (also called
gamma hemolysis).

Sheep Blood Agar

Sheep Blood Agar - uninoculated

Type: Differential and enriched

Purpose: Determine types of hemolysis (i.e., α, β, γ)

Interpretation: α: partial clearing, green or brownish ring; β: wide zone of clearing; γ: non-hemolytic

Examples

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