Growth & Culture of Bacteria. Chapter 4, 5

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1 Growth & Culture of Bacteria Chapter 4, 5

2 Key Words Obligate aerobe Obligate anaerobe Facultative anaerobe Microaerophilic Generation time Growth curve Mesophile Thermophile Psychrophile 2

3 Growth of bacteria Bacterial Multiplication - Binary Division 1 to 2 to 4 to 8 to N Log Growth? Chapter 4

4 Chapter 4 Fig. 4-2 Phase Lag Exponential Maximum stationary Decline Growth Rate Zero Constant Zero Negative (death)

5 Phases of bacteria growth (in vitro) Growth curve Lag Adapt to nutrients Log Active growth Stationary Death = Growth rate Death Nutrients consumed ph too low (why?) Optimize curves in production

6 Note the drop-off as you get closer to the Optimum!

7 Bacterial Growth Curve Is it the same as in human body? Why?

8 Nutritional Factors Carbon Source Hetero- or Auto- Organic compounds or CO 2 Nitrogen Source NO 3 - ==> NO 2 ==> NH 3 to Amino Acids ==> Proteins Sulfur Phosphorus Trace Elements Fe, some required for growth or virulent Host sideophores bind Fe

9 All pathogenic bacteria are heterotrophic -obtain energy by oxidizing organic molecules (Carbohydrates, lipids and protein) Metabolism- yield ATP as an energy source -Aerobic respiration, 1 glucose produce 38 ATP - enzymes -Anaerobic respiration, 1 glucose produce 2 ATP (fermentation) -enzymes Requirement for oxygen in respiration -obligate -facultative Cultivation of bacteria Chapter 5 p.657

10 Requirements for bacterial growth Nutrients H 2 O, C-source(organic), N-source (organic) Inorganic salts Growth factors Temperature ph Gas Pathogenic bacteria culture condition? Incubator Temperature, gas Culture medium Nutrients, ph

11 Moisture Salt concentration 0.9% Nacl PBS(Phosphate buffer saline)=? Isotonic Osmotic pressure Pseudomonas grows in distilled water Jelly- Jams (high sugar) Salt pork (high salt)

12 Temperature Requirements Mesophile 37 (Pathogenic bacteria) Thermophiles Hot spring, hot tubs-public Heat Resistant Enzymes in PCR (Tag DNA polymerase) Psychrophiles Cold food (growth in transit) Listeria grows in hot dogs

13 What effect would have a fever on Mesophiles?

14 Hydrogen Ion concentration (ph) Neutralophiles (ph ) Acidophiles (< ph 3.0) (Helicobacter pylori) Alkaliphiles (ph10.5) (bacteria from salina, Vibrio. cholera) Microorganismis regulate their internal ph over a wide range of external ph values by pumping protons in or out of the cell Chapter 6

15 Oxygen Requirements Classification Aerobic Anaerobic Falcultative Anaerobic Microaerophilic O 2 is reactive-toxic (binds e- s) Oxidization of compounds Loss of energy from compounds DNA molecule breaks

16 Why can some organisms grow in the presence of oxygen? Toxic forms of oxygen need to be neutralized by enzymes -Superoxide dismutase -Catalase -Peroxidase If microbe does not produce these enzymes must have anaerobic conditions

17 . Culture medium -the mixture of various nutrients that is suitable for the growth of microorganisms Types of Culture Media -based on the function and chemical components -based on the physical state

18 Based on the function and the chemical components Basic Medium -contains the basic nutrients for the most bacterial growth; -the base of other kind of media. e.g. broth. Nutrient Medium/Enriched Medium Additional or special nutrients (e.g., serum, growth factors, trace elements) are added to support some fastidious bacterial growth. e.g. blood agar. 18

19 Selective Medium the medium that can prevent the certain bacterial growth while permitting others. e.g. SS agar Differential Medium Some special substrates and indicators are added into the media in order to produce a visual differentiation when several bacteria grow on the same kind of medium. e.g. EMB agar (Eosin-methylene blue agar). 19

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22 Review questions (chapter 2) 1. p41:1, 2, 3, 5, 6, 7, 8, 9 and think about explainations. 2. Summary the differences between the cell wall of G + bacteria and G - bacteria (related with clinical practice) 3. Why penicillin and lysozyme have less effect on G- bacteria? 4. Structures of bacteria associated with clinical practice. Explain which bacterial structures can be as an antibiotics target.

23 Review questions(chapter 4 & 5) 1. In p65: question 1, is it right? Why? 2. In p65: question 2, 4, 5 3. Why a doctor should know the growth curve of bacteria? 4. In p76: 1, 3, 5, 10 Ref. Medical microbiology Published by Mosby ed. MIMS, PLAYFAIR, ROITT WAKELIN, WILLIAMS Review of Medical Microbiology and Immunology -AccessMedicine:

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