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Bacterial Requirements

Bacterial Requirements. Temperature and Oxygen. 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. Metabolic Classifications.

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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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