Types of culture media Flashcards by Coleen Trixie Simpliciano

Culture

Are growth of microorganism on a culture medium

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Is a liquid, semi-solid, or solid medium utilized to observe growth patterns of microorganism as well as for transport and storage

Culture medium

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Types of culture (3)

Pure
Mixed
Stock

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Pure culture

Composed only one specie

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Mixed culture

Composed of more than one specie

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Composed of several species contained in a separate culture medium
Should be grown in large volume of broth
Used for academic and industrial purposes

Stock culture

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Classification of culture medium according to consistency (3)

-Liquid medium
-Semi-solid medium
-Solid medium

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Contains 0% agar
Allows growth of aerobes, anaerobes and facultative anaerobes

Liquid medium

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3 Examples of liquid medium

-Brain heart infusion
-Trypticase Soy Broth
-Thioglycollate

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Semi-solid medium

Contains 0.5-1% agar

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Example of semi-solid medium

Sulfide Indole Motility

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Solid medium

Contains 2-3% agar

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4 Examples of solid medium

-Triple Sugar Iron (TSI) agar
-Mac Conkey
-Blood Agar Plate
-Chocolate Agar Plate

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Is a sulfated polymer containing D-galactose, D-glucuronic acid; derived from red algae

Agar

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Agar will melt if the temperature reaches

80-90

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Agar will solidify if the temperature reaches

40-50

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Cooling temperature for distribution of agar into Petri dish

55-60

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Classification of culture medium according to composition (3)

-Synthetic/defined Medium
-Non-synthetic/complex medium
-Tissue Culture Medium

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Medium where all components are known to the user
Used for research purposes

Synthetic/Defined medium

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Non-synthetic/Complex medium

-Composed of some unknown substances
-Useful for isolation of bacteria

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3 Examples of non-synthetic medium

-Nutrient Broth (NB)
-Trypticase Soy Broth (TSB)
-Mac Conkey agar

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Used for obligate intracellular bacteria (Rickettsia & Chlamydia)

Tissue culture medium

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Classification of culture medium according to how the medium is dispensed/distributed (2)

-Plated medium
-Tube medium

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Tube medium (4

Liquid
Butt
Buttslant
Slant

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Classification of culture medium according to use (6)

-Simple Media/General Purpose ---Media/Supportive Media -Enrichment media -Enriched media -Differential media -Selective media Special media

Routinely used in the lab and w/o added supplement Media that support the growth of most nonfastidious bateria

-Simple Media/General Purpose -Media/Supportive Media

Composition of Simple Media/General Purpose Media/Supportive Media

Meat & soybean extracts

2 Examples of Simple Media/General Purpose Media/Supportive Media

-Nutrient agar (NA) and broth (NB) -Trypticase Soy Broth

Used to propagate growth of certain group of bacteria from a mixture of organisms Contains specific type of nutrients Liquid-type media

Enrichment media

Media with added supplements necessary for the growth of fastidious organisms Solid-type media

Enriched/non-selective media

Standard nonselective medium

Blood Agar Plate (BAP)

Is with horse blood; used for isolation of hemophilus

Chocolate Agar Plate (CAP)

Media that allow visualization of metabolic differences between groups of bacteria

Differential media

Differentiates lactose fermenters (pink colonies) from nonlactose fermenters (colorless colonies)

Mac Conkey

Differentiates hemolytic pattern & what hemolytic pattern?

-BAP -Streptococci

Hemolytic patterns (3)

-Alpha -Beta -Gamma

Greenish zone Partial/increased hemolysis

Alpha

Clear zone Complete lysis of RBCs around colony

Beta

No hemolysis No change in agar

Gamma

Media incorporated with antibiotics, dyes or chemicals to inhibit the growth of undesired organisms while promoting the growth of desired organism

Selective media

6 Examples of selective media

-Hektoen enteric (HE) agar -Mac Conkey agar -Xylose Lysine Desoxycholate (XLD) agar -Bismuth sulfite agar (BSA) -Mannitol salt agar -Thayer Martin agar (TMA)

Gentamicin Blood Agar

Streptococci

Bacitracin Chocolate Agar

Hemophilus

Blood Agar Plate with Ampicillin

Aeromonas

Phenol Ethyl Alchohol

Gram-positive cocci

Columbia agar with CNA

Gram-positive bacteria

Gram-negative (GN) Broth

-Salmonella spp. -Shigella spp.

Thayer Martin Agar (TMA)

Neisseria gonorrheae

Special media

Used to isolate bacteria with specific growth requirements

2 Examples of special media

-Lowenstein Jensen (LJ) medium -Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar

Red with black centers

Salmonella

Red or clear colonies without black centers

Shigella

Used as a filler for the remaining percentage of the anaerobic atmosphere

Nitrogen gas

Remove residual oxygen from the chamber by combining with hydrogen to form water

Palladium pellets

Used to absorb the water formed when hydrogen combines with free oxygen in the presence of the catalyst

Silica gel (dessicant)

An oxygen reduction indicator; it becomes colorless in the absence of oxygen

Methylene blue or reazurin

Inoculation techniques (5)

-Streaking -Placement if fluid specimens or swabs into culture broth -Stabbing the medium -Stab and streak technique -Pour plate technique

Most commonly performed inoculatin technique

Streaking

Streaking (5)

-Radial streak -Overlap streak -Multiple streak -Multiple interrupted -Interrupted

The microorganisms present in the specimen are successively diluted out as each quadrant is streaked until finally each morphotype is present as a single colony

Streaking fir isolation

This technique is applied to semi-solid medium like SIM medium Also used to inoculate streptococci into BAP

Stabbing

Used to inoculate organism into the butt slant tube medium such as the TSI agar and LIA

Stab and streak

Is a technique where the organism is added to a molten agar, mixed and then allows the medium to solidify

Pour plate technique

pH indicators (4)

Phenol red Bromthymol blue Neutral red Bromcresol purple

Bromthymol blue

HEA SCA TCBS Malonate broth

Phenol red

TSI XLD MSI Christensen's urea sugar

Neutral red

Mac Conkey SSA

Bromcresol purple

LIA

Neutral Red + Acid

Red

Neutral Red + Alkaline

Yellow

Phenol Red + Acid

Yellow

Phenol Red + Alkaline

Red

Bromthymol Blue + Acid

Orange to Yellow

Bromthymol Blue + Alkaline

Green to blue

Thosulfate-Citrate-Bile salt Sucrose (TCBS) inhibitor, Ph Indicator, and Differentiating factor

Bile salt Bromthymol blue Sucrose

Salmonella Shigella (SS) Agar inhibitor, Ph Indicator, and Differentiating factor

Bile salt Neutral red Lactose, H2S

MacConkey (MAC) Agar inhibitor, Ph Indicator, and Differentiating factor

Crystal violet Neutral red Lactose

Hektoen Enteric Agar (HEK, HE or HEA) inhibitor, Ph Indicator, and Differentiating factor

Bile salt Bromthymol blue Lactose, sucrose, salicin, H2S

Xylose Lysine Deoxycholate Agar (XLD) inhibitor, Ph Indicator, and Differentiating factor

Bile salt Phenol red Lactose, sucrose, xylose, H2S

Eosin Methylene Blue Agar (EMB) inhibitor, Ph Indicator, and Differentiating factor

Eosin Y/ MB Eosin Y/ MB Lactose

Small zone of intact RBCs surrounded by a zone of Beta hemolysis

Inner zone of beta hemolysis surrounded by a zone of alpha

Double