lab 3 culture media 2012
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Second year Lab Medicine (Females)1433- 1434 A.H. / 2012- 2013
Microbiology Practical (Course I)
Mrs. Hibah Abdul-Wahab I. Abu-Sulaiman
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Who will be her groom??!
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Royal Dining Royal Atmosphere
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Bacterial nutritional requirements. Composition of culture media. Categories of culture media. Solidifying agents. Types of media. Media Quality control.
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1. Carbon Source:◦ Heterotrphs: from organic compounds such as:
a.a., polysaccharides, CHO, proteins and peptides.
◦ Autotrophs: from fixing inorganic carbon dioxide.
2. Energy Source:◦ Phototrophs: from sunlight.◦ Chemotrophs: from oxidation of chemicals.
3. Nitrogen: essential for a.a., nucleotide and vitamins synthesis.
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4. Minerals: as cofactors in enzymatic reactions.◦ E.g., sodium, potassium, calcium, magnesium, iron…
etc.5. Growth factors:
◦ Amino acids, a.a.: bacteria possess proteinases.◦ Purines and pyrimidines: to be converted to
nucleotides.◦ Bacterial vitamins: for co-enzyme production.
6. Water: ◦ All metabolic functions of a bacterial cell are
performed in aqueous environment.◦ Distilled water
Tap water minerals and their reaction with peptones and meat extracts.
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1. Water.2. Peptone: product of protein hydrolysis.
◦ From animal proteins: source of nitrogen.◦ From soya bean (plant) protein: source of
carbohydrates, CHO.
3. Meat extracts: as source of a.a. and other essential vitamins and elements.
4. Yeast extract: as a stimulant for bacterial growth in the media.
5. Mineral salts: for bacterial enzymes activity.6. Carbohydrates: as source of carbon and
energy.
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Agar:◦ Is a complex polysaccharide (polymer of
galactose) that is extracted from the cell wall of seaweed (red algae).
◦ It melts at 100oC and at 37oC it is solid.◦ It has no nutritional value in agar cultures.◦ It is added in a concentration of 1.5% to solidify
media and 0.4% to make semisoft media. Gelatin:
◦ A protein which is derived from collagen of skin and bone.
◦ Some bacteria are able to liquefy gelatin at 37oC.
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Without agar
With agar
In the form of Can be in the form of
Primarily used for Primarily used for
Consist of
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Agar Slant Agar Deep Agar Plates
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In broths, bacterial growth is indicated by a change in the appearance from clear to turbid (cloudy).
Turbidity of the broth is due to light deflected by bacteria present in the culture.
The more turbid, the….
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Basal Media:◦ Simple media that will support the growth of most bacteria
that do not require special nutrient. E.g., Nutrient broth.◦ Nutrient broth+ agar= Nutrient agar
Enriched media:◦ Culture media that are enriched with whole or lysed blood,
serum, special extracts or nutrient to support the growth of those bacteria that cannot grow on the basal media.
◦ Nutrient agar+ blood= Blood agar◦ Blood agar+ heat= chocolate agar
Selective media:◦ Solid media which contain substances that inhibit the growth
of some bacteria and letting the pathogenic ones grow.
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Enrichment media:◦ These are liquid media that are similar in function
to ...◦ The only difference is that…
Differential media:◦ Contains indicators that will differentiate one
organism from another Transport media:
◦ Contain substances that can prevent overgrowth of commensals and prevent bacteria from dying as a result of change in pH or enzyme action.
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Media should be tested for:1. Sterility.2. Each medium must be tested with organisms
expected to grow or give a positive reaction as well as with organisms expected not to grow.
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In a table, define each of the following media sugar, inhibitors, indicators and other additives:1. Bile Esculin Agar (BEA).2. Blood Agar.3. Chocolate Agar.4. Cysteine-tellurite Agar.5. MacConkey Agar.6. Mannitol Salt Agar.7. Salmonella-Shigella Agar.8. Selenite broth.9. Thayer-Martin Agar.10.Thiosulfate Citrate-bile salts (TCBS) Agar.11.Xylose lysine deoxycholate (XLD) agar.
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