1. Growth Media and Parameters

2. Complex Media Made from complex and rich ingredients –Ex. Soya protein extracts –Milk protein extracts –Blood products –Tomato juice, etc. Exact chemical composition unknown Can be selective and/or differential 2

3. Defined Media Known chemical composition – May contain up to 80 different ingredients – May be quite simple – Allows the growth of a restricted number of microorganisms – Highly variable composition as a function of the microorganism May be selective and/or differential 3

4. Selective Media Contains compounds which inhibit or kill the unwanted organisms –Ex. Medium containing penicillin only allows the growth of penicillin resistant microorganisms 4

5. Differential Media 5 Allows the discrimination of different species Often contain pH indicators –Allows the discrimination of different metabolisms Production of acid products turns the medium yellow Production of alkaline products turns the medium red

6. Nutrition Macronutrients –C,H,N,O,P,S

7. Carbon Required for the synthesis of all organic compounds : –Carbohydrates –Lipids –Proteins –Nucleic acids

8. Organic –Monosaccharides –Disaccharides –Polysaccharides –Proteins –Lipids –Nucleic acids –Phenols, Etc. Inorganic –CO 2 –CO Carbon Sources

9. Phosphorous Required for the synthesis of : –Nucleic acids –Phospholipids –ATP –Used as a buffer; control of pH Sources: –Organic and inorganic The inorganic form is the most used

10. Nitrogen Required for the synthesis of: –Amino acids –Nucleic acids –Peptidoglycan Sources: –Organic: Amino acids –Inorganic: NH 3, NO 3, & N 2

11. Sulfur Required for the synthesis: –Amino acids (Cysteine/Methionine) –Vitamins (thiamine and biotin) Sources: –Organic: Amino acids Cystein and methionine –Inorganic: S, SO 4

12. Hydrogen and Oxygen Required for the synthesis of organics!! –Carbohydrates –Lipids –Proteins –Nucleic acids Sources: –Organic: Organic carbon –Inorganic: H 2 (Methanogens only)

13. Nutritional Classification Carbon sources –Heterotrophs: Preformed organic compounds –Autotrophs: Inorganic molecules –CO 2 and CO

14. Nutritional Classification (Cont’d) Energy sources –Phototrophs: Light –Chemotrophs: Oxidation of organic and inorganic compounds Source of e- –Organotrophs: Reduced organic molecules –Lithotrophs: Reduced inorganic molecules

15. Nutritional Types Autotrophs photolithotrophs Heterotrophs photoorganotrophes Autotrophs chemolithotrophs Heterotrophs chemoorganotrophs

16. What you have to know about the media What are the sources of C,H,N,O,P,S? What type of media is it? What are the indicators? What are the selective agents? They allow the growth of what bacteria? What are the possible reactions?

17. Ex. MacConkey Agar Peptone - 17 g Proteose peptone - 3 g Lactose - 10 g Bile salts - 1.5 g Sodium chloride - 5 g Neutral red - 0.03 g Crystal Violet - 0.001 g Agar - 13.5 g Sources of C,H,N,O,P,S? Type of media? Indicators? Selective agents? Allow growth of what bacteria? Possible reactions?

18. Environmental Parameters Oxygen availability pH Temperature 18

19. Oxygen Requirements Aerobic: –Absolute need of oxygen to survive –Used as a final electron acceptor –Used by bacteria that carry out an oxidative or aerobic respiratory metabolism Microaerophilic: –Absolute need for low concentrations of oxygen –High concentrations are detrimental

20. Oxygen Requirements (Cont’d) Anaerobic/Aerotolerant: –Oxygen is not required for growth or survival but is tolerated Facultative anaerobes: – Facultative oxygen requirement – May use oxygen or not – Possesses an oxygen dependant and oxygen independent metabolism Strict or obligate anaerobic : –Oxygen is neither used nor tolerated; cannot survive in the presence of oxygen