Download - Anaerobic Culture Methods
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Anaerobic Culture Methods
• Reducing media– Contain chemicals (thioglycollate or oxyrase)
that combine O2
– Heated to drive off O2
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Anaerobic Culture Methods
• Anaerobic jar
Figure 6.5
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Anaerobic Culture Methods
• Anaerobic chamber
Figure 6.6
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Capnophiles Require High CO2• Candle jar
• CO2-packet
Figure 6.7
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Selective Media
• Suppress unwanted microbes and encourage desired microbes.
Figure 6.9b–c
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Differential Media
• Make it easy to distinguish colonies of different microbes.
Figure 6.9a
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Enrichment Media
• Encourages growth of desired microbe
• Assume a soil sample contains a few phenol-degrading bacteria and
thousands of other bacteria
– Inoculate phenol-containing culture medium with the soil and
incubate
– Transfer 1 ml to another flask of the phenol medium and incubate
– Transfer 1 ml to another flask of the phenol medium and incubate
– Only phenol-metabolizing bacteria will be growing
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• A pure culture contains only one species or strain.
• A colony is a population of cells arising from a single cell or spore or from a group of attached cells.
• A colony is often called a colony-forming unit (CFU).
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Streak Plate
Figure 6.10a–b
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Preserving Bacteria Cultures
• Deep-freezing: –50°to –95°C
• Lyophilization (freeze-drying): Frozen (–54° to –72°C) and dehydrated in a vacuum
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Reproduction in Prokaryotes
• Binary fission
• Budding
• Conidiospores (actinomycetes)
• Fragmentation of filaments
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Binary Fission
Figure 6.11
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Figure 6.12b
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• If 100 cells growing for 5 hours produced 1,720,320 cells:
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Figure 6.13
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Figure 6.14
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Direct Measurements of Microbial Growth
• Plate counts: Perform serial dilutions of a sample
Figure 6.15, step 1
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Plate Count
• Inoculate Petri plates from serial dilutions
Figure 6.16
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Plate Count• After incubation, count colonies on plates that have
25-250 colonies (CFUs)
Figure 6.15
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Direct Measurements of Microbial Growth
• Filtration
Figure 6.17
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Direct Measurements of Microbial Growth
• Multiple tube MPN test.
• Count positive tubes and compare to statistical MPN table.
Figure 6.18b
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Direct Measurements of Microbial Growth
• Direct microscopic count
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Direct Measurements of Microbial Growth
Figure 6.19, steps 1, 3
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Estimating Bacterial Numbers by Indirect Methods
• Turbidity
Figure 6.20
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Measuring Microbial GrowthDirect methods
• Plate counts
• Filtration
• MPN
• Direct microscopic count
• Dry weight
Indirect methods
• Turbidity
• Metabolic activity
• Dry weight