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Growth Media and Parameters 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 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 Selective Media • Contains compounds which inhibit or kill the unwanted organisms – Ex. Medium containing penicillin only allows the growth of penicillin resistant microorganisms 4 Differential Media • Allows the discrimination of different species • Often contain pH indicators – Allows the discrimination of different metabolisms Production of alkaline products turns the medium red Production of acid products turns the medium yellow 5 Nutrition • Macronutrients – C,H,N,O,P,S Carbon • Required for the synthesis of all organic compounds : – – – – Carbohydrates Lipids Proteins Nucleic acids Carbon Sources • Organic – – – – – – – Monosaccharides Disaccharides Polysaccharides Proteins Lipids Nucleic acids Phenols, Etc. • Inorganic – CO2 – CO 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 Nitrogen • Required for the synthesis of: – Amino acids – Nucleic acids – Peptidoglycan • Sources: – Organic: Amino acids – Inorganic: NH3, NO3, & N2 Sulfur • Required for the synthesis: – Amino acids (Cysteine/Methionine) – Vitamins (thiamine and biotin) • Sources: – Organic: Amino acids • Cystein and methionine – Inorganic: • S, SO4 Hydrogen and Oxygen • Required for the synthesis of organics!! – – – – Carbohydrates Lipids Proteins Nucleic acids • Sources: – Organic: • Organic carbon – Inorganic: • H2 (Methanogens only) Nutritional Classification • Carbon sources – Heterotrophs: • Preformed organic compounds – Autotrophs: • Inorganic molecules – CO2 and CO 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 Nutritional Types • • • • Autotrophs photolithotrophs Heterotrophs photoorganotrophes Autotrophs chemolithotrophs Heterotrophs chemoorganotrophs 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? Ex. MacConkey Agar • • • • Sources of C,H,N,O,P,S? • Type of media? • Indicators? • Selective agents? Allow growth of what bacteria? • • • • • • • Possible reactions? 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 Environmental Parameters • Oxygen availability • pH • Temperature 18 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 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