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GROWTH AND CULTURING OF BACTERIA

GROWTH AND CULTURING OF BACTERIA. CHAPTER 6. Growth and Cell Division. Growth Binary fission. Cell Division. Budding. Bacterial Growth Curve. Lag phase Log phase Stationary phase Death phase. Measuring Bacterial Growth. Serial dilution/ Standard plate counts.

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GROWTH AND CULTURING OF BACTERIA

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  1. GROWTH AND CULTURING OF BACTERIA CHAPTER 6

  2. Growth and Cell Division • Growth • Binary fission

  3. Cell Division • Budding

  4. Bacterial Growth Curve • Lag phase • Log phase • Stationary phase • Death phase

  5. Measuring Bacterial Growth • Serial dilution/ Standard plate counts

  6. Measuring Bacterial Growth • Direct microscopic counts

  7. Measuring Bacterial Growth • Most probable number

  8. Measuring Bacterial Growth • Filtration • Turbidity (Spectrophotometric)

  9. Filtration

  10. Turbidity

  11. Physical Factors Affecting Bacterial Growth • pH • Temperature

  12. Physical Factors Affecting Bacterial Growth • Oxygen • Moisture

  13. Physical Factors Affecting Bacterial Growth • Hydrostatic pressure • Osmotic pressure • Radiation

  14. Radiation: Ionizing (gamma radiation) Ultraviolet radiation: DNA absorbs 260 nm light “Thymine dimers”

  15. Nutritional Factors Affecting Bacterial Growth • Carbon source • Nitrogen source • NH4 best, but NO3 and NO2 can work too. • Sulfur/ phosphorus C6H12O6 best but many sugars CO2 for autotrophs SO42- PO43-

  16. Nutritional Factors Affecting Bacterial Growth • Trace elements • Vitamins • Nutritional complexity

  17. Locations of Enzymes • Extracellular • Periplasmic • kinases and phosphatases • Cytoplasmic Proteases and cellulases (hydrolases) Foldases , dehydrogenases

  18. Sporulation • Endospores • Other sporelike bacterial structures

  19. Bacillus subtilis spore stain

  20. Culturing Bacteria — Streak Plates • Streak plate method

  21. Culturing Bacteria — Pour Plates • Pour plate method

  22. Culture Media • Types • Defined • Complex LB (Luria-Bertani) TSA

  23. Culture Media Purposes • Selective media • Differential media • Enrichment media McConkey’s

  24. Detection of Carbon source fermenters McConkey’s agar

  25. Culture Conditions — O2 • Controlling oxygen content

  26. Culture conditions • Maintaining cultures • Preserved cultures • Diagnostic tests • Limitations

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