blood agar

Agar plate

An agar plate streaked with microorganisms isolated from a deep-water sponge. Individual colonies may be seen center right.
Uses	Culturing
Related items	Petri dish Growth medium

An agar plate is a sterile Petri dish that contains a growth medium (typically agar plus nutrients) used to culture microorganisms or small plants like the moss Physcomitrella patens.

Selective growth compounds may also be added to the media, such as antibiotics.^[1]

Individual microorganisms placed on the plate will grow into individual colonies, each a clone genetically identical to the individual ancestor organism (except for the low, unavoidable rate of mutation). Thus, the plate can be used either to estimate the concentration of organisms in a liquid culture or a suitable dilution of that culture using a colony counter, or to generate genetically pure cultures from a mixed culture of genetically different organisms, using a technique known as "streaking". In this technique, a drop of the culture on the end of a thin, sterile loop of wire, sometimes known as an inoculator, is streaked across the surface of the agar leaving organisms behind, a higher number at the beginning of the streak and a lower number at the end. At some point during a successful "streak", the number of organisms deposited will be such that distinct individual colonies will grow in that area which may be removed for further culturing, using another sterile loop.^[1]

Types of agar plates

An agar plate being viewed in an electronic colony counter.

Like other growth media, the formulations of agar used in plates may be classified as either "defined" or "undefined"; defined medium is synthesized from individual chemicals required by the organism so that the exact molecular composition is known, whereas undefined medium is made from natural products such as yeast extract, where the precise composition is unknown.^[2]

Agar plates may be formulated as either permissive, with the intent of allowing the growth of whatever organisms are present, or restrictive or selective, with the intent of only allowing growth a particular subset of those organisms.^[3] This may take the form of a nutritional requirement, for instance providing a particular compound such as lactose as the only source of carbon and thereby selecting only organisms which can metabolize that compound, or by including a particular antibiotic or other substance in order to select only organisms which are resistant to that substance. This correlates to some degree with defined and undefined media; undefined media, made from natural products and containing an unknown combination of very many organic molecules, is typically more permissive in terms of supplying the needs of a wider variety of organisms, while defined media can be precisely tailored to select organisms with specific properties.

Agar plates may also be indicator plates, in which the organisms are not selected on the basis of growth, but are instead distinguished by a color change in some colonies, typically caused by the action of an enzyme on some compound added to the media.

Some commonly used agar plate types are:

Red blood cells on an agar plate are used to diagnose infection. On the left is a positive Staphylococcus infection, on the right a positive Streptococcus culture.

Blood agar types

• Blood agar plate (BAP)

Contains mammalian blood (usually sheep or horse), typically at a concentration of 5–10%. BAP are an enriched, differential media used to isolate fastidious organisms and detect hemolytic activity. β-hemolytic activity will show complete lysis of red blood cells surrounding colony. Examples include Streptococcus haemolyticus. α-hemolysis will only partially lyse hemoglobin and will appear green. An example of this would be Streptococcus viridans. γ-hemolysis (or non-hemolytic) is the term referring to a lack of hemolytic activity.

Contains meat extract, tryptone, sodium chloride, and agar.

• Chocolate agar (CHOC)

A type of blood agar plate in which the blood cells have been lysed by heating the cells to 56 °C. Chocolate agar is used for growing fastidious (fussy) respiratory bacteria, such as Haemophilus influenzae. No chocolate is actually contained in the plate; it is named for the coloration only.

• Thayer-Martin agar (TM)

Chocolate agar designed to isolate Neisseria gonorrhoeae.

• TCBS

TCBS (thiosulfate citrate bile salts sucrose)-enriched agar enhances growth of Vibrio cholerae ^[4]

General bacterial media

Four types of agar plates demonstrating differential growth depending on bacterial metabolism.

• Bile esculin agar (BEA)

BEA is used for the isolation of Enterococci as well as Group D Streptococci

• CLED agar (Cysteine Lactose Electrolyte Deficient agar)

CLED agar is used to isolate and differentiate urinary tract bacteria, since it inhibits Proteus species swarming and can differentiate between lactose fermenters and non-fermenters.

• Hektoen enteric agar (HEA)

HE agar is designed to isolate and recover fecal bacteria belonging to the Enterobacteriaceae family. HE is particularly useful in isolating Salmonella and Shigella.

• Lysogeny Broth (LB)
• McConkey agar (MAC)

A selective and differential media used to differentiate between Gram negative bacteria while inhibiting the growth of Gram positive bacteria. The addition of bile salts and crystal violet to the agar inhibits the growth of most Gram positive bacteria, making MacConkey agar selective. Lactose and neutral red are added to differentiate the lactose fermenters, which form pink colonies, from lactose nonfermenters that form clear colonies. An alternative media, eosin methylene blue (EMB) serves a similar purpose.

• Mannitol salt agar (MSA)

MSA is also a selective and differential media. The mannitol indicates organisms that ferment mannitol: mannitol fermentation produces lactic acid, lowering the pH and turning the plate yellow. The salt is to select for halophiles; organisms that cannot withstand a high salt content will be unable to grow well.

• Mueller-Hinton agar (MHA)

MHA contains beef infusion, peptone, and starch and is used primarily for antibiotic susceptibility testing. It can be in a form of blood agar.

Fungi growing in axenic culture (ascomycetes)

• Nutrient agar

Nutrient agar is usually used for growth of non-fastidious organisms and observation of pigment production. It is safe to use in school science laboratories because it does not selectively grow pathogenic bacteria.

• Önöz agar

Önöz agar allows more rapid bacteriological diagnosis as Salmonella and Shigella colonies can be clearly and reliably differentiated from other Enterobacteriaceae. The yields of Salmonella from stool samples obtained, when using this medium, are higher than those obtained with LEIFSON Agar or Salmonella–Shigella agar (SSA).

• Phenylethyl alcohol agar (PEA)

PEA selects for Staphylococcus species while inhibiting Gram-negative bacilli (e.g. Escherichia coli, Shigella, Proteus, etc.).

Aspergillus niger growing in potato dextrose agar

• R2A Agar (R2A)

A non-specific agar that imitates the medium of water. Used for water analysis.

• Tryptic (Trypticase) Soy Agar (TSA)

TSA is a general purpose media produced via enzymatic digestion of soybean meal and casein. TSA is frequently the base media of other agar types; for example, blood agar plates are made by enriching TSA plates with blood (see above).

TSA plates support growth of many semi-fastidious bacteria, including some species of Brucella, Corynebacterium, Listeria, Neisseria, and Vibrio.

• Xylose-Lysine-Deoxycholate agar (XLD)

XLD is used for the culture of stool samples and contains two indicators. It is formulated to inhibit Gram-positive bacteria, while the growth of Gram-negative bacilli is encouraged. The colonies of lactose fermenters appear yellow.

It is also used to culture possible Salmonella that may be present in a food sample. Salmonella colonies will show a black halo on XLD.

• Cetrimide agar

Cetrimide agar is a type of agar used for the selective isolation of the gram-negative bacteria, Pseudomonas aeruginosa.

• Tinsdale agar contains potassium tellurite, which can isolate Corynebacterium diphteriae. ^[4]

Fungal media

• Sabouraud agar

Sabouraud agar is used to culture fungi and has a low pH that inhibits the growth of most bacteria; it also contains the antibiotic gentamicin to specifically inhibit the growth of Gram-negative bacteria.

• Hay infusion agar

Specific for the culturing of slime moulds (which are not fungi).

• Potato dextrose agar

PDA is used to culture certain types of fungi.

• Malt extract agar

Malt extract agar has a high content of peptone and is acidic. It is essentially used in the isolation of fungal microorganisms.

Moss media

Physcomitrella patens plants growing axenically on an agar plate (Petri dish, 9 cm diameter).

• Knop agar

Knop agar is used to culture axenically protonema and whole moss plants, e.g. those of Physcomitrella patens, a model organism. ^[5]

History

In 1881, Fannie Hesse, the wife of a technician in the laboratory of Robert Koch, suggested agar as an effective setting agent since it had been common place in jam making for some time.^[6]

References

See also

Wikimedia Commons has media related to: Agar plate

• Growth medium
• MRS agar
• Petri dish
• R2a agar
• Replica plating
• Viral plaque

External links

• Video of sheep blood agar preparation
• Science Buddies - All About Agar