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Bacterial Requirements

This article covers the measurement of microbial growth, temperature requirements, oxygen requirements, and metabolic classifications of bacteria. It also includes a temperature experiment and anaerobic culture methods.

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Bacterial Requirements

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  1. Bacterial Requirements Temperature and Oxygen

  2. Measurement of Microbial Growth • CFU • Direct • Viable Plate Counts • Serial Dilutions • Pour Plate • Spread Plate • Membrane Filtration • Counting Chamber • MPN (fecal, H20, food) • Indirect • Turbidity • Metabolic activity

  3. Metabolic Classifications • Temperature • Oxygen requirements • pH • Pressure • Radiation

  4. Growth Curve Labeled Phases • Lag phase • Adaptive • Start metabolism • 1-3 days • Log phase • Generation time doubles • Most metabolically active • Stationary phase • Growth = death • Death phase • Requirements decrease • Possible spore formation

  5. Temperature Requirements

  6. Temperature Experiment • 4 TSA plates • Label • Temperature • 4 different settings • Wheel spoke pattern • 5 Bacteria • 4 SAB broth/Durham tube • Temperature • 4 different settings • S. cerevisiae

  7. Anaerobic Culture Methods

  8. Oxygen Requirements • Obligate Aerobes • Microaerophiles • Aerotolerant anaerobes • Obligate Anaerobes • Facultative Anaerobes • Capnophiles

  9. BHI deeps • Brain/Heart infusion • Melt • Label • BHI • Organism • Inoculate • Pipette or loop • Roll tube to distribute • Ice Bath • Incubate @ 37°C

  10. Fluid Thioglycollate Broth • Sodium thyioglycollate • Reduction (redox) • Pink if oxidized • Label tubes • Thio • Organism • Inoculate with loop • Incubate @ 37°C

  11. Gas Pack Jar: CO2 anaerobic System TSA agar plate 4 quadrants Label bacteria Label O2 H2O added to envelope to Produce H2 and CO2 NaHCO3, NaBH4 H2 + O2 => H2O

  12. Aerobic / Anaerobic Lab Test Results aerobe FA aerotolerant anaerobe

  13. Physical Requirements: Temperature • Psychrophile • Unsaturated FA in cell membrane • Cold temperatures • Psychrotroph (facultative psychrophiles) • Refrigeration • Room temperature • Mesophile • Warm Blooded Animals • Thermophile/Hyper- • Saturated FA in cell membrane

  14. Bacterial Examples: Temperature psychrophile psychrotroph mold Vibrio mesophile thermophile Archaea E coli

  15. Oxygen Requirement Classification

  16. BHI results

  17. Thio Tubes

  18. Bacterial Examples of O2 Groups • Obligate Aerobes • Pseudomonas • Microaerophiles • H. pylori • Aerotolerant aerobes • Streptococcus • Lactobacillus • Obligate Anaerobes • Clostridium • Facultative Anaerobes • E. coli • Stapylococcus • Capnophilic • Campylobacter E coli Staph

  19. pH Groups • Acidophile • Bacillus acidocaldarius • Lactobacillus acidophilus • Neutrophile • E. coli • Staphylococcus aureus • Akaliphile • Streptococcus pneumoniae • Nitrobacter sp.

  20. Questions?

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