1. Anaerobic Culture Methods Reducing media –Contain chemicals (thioglycollate or oxyrase) that combine O 2 –Heated to drive off O 2

2. Anaerobic Culture Methods Anaerobic jar Figure 6.5

3. Anaerobic Culture Methods Anaerobic chamber Figure 6.6

4. Capnophiles Require High CO 2 Candle jar CO 2 -packet Figure 6.7

5. Selective Media Suppress unwanted microbes and encourage desired microbes. Figure 6.9b–c

6. Differential Media Make it easy to distinguish colonies of different microbes. Figure 6.9a

7. 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 –Only phenol-metabolizing bacteria will be growing

8. 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).

9. Streak Plate Figure 6.10a–b

10. Preserving Bacteria Cultures Deep-freezing: –50°to –95°C Lyophilization (freeze-drying): Frozen (– 54° to –72°C) and dehydrated in a vacuum

11. Reproduction in Prokaryotes Binary fission Budding Conidiospores (actinomycetes) Fragmentation of filaments

12. Binary Fission Figure 6.11

13. Figure 6.12b

14. If 100 cells growing for 5 hours produced 1,720,320 cells:

15. Figure 6.13

16. Figure 6.14

17. Direct Measurements of Microbial Growth Plate counts: Perform serial dilutions of a sample Figure 6.15, step 1

18. Plate Count Inoculate Petri plates from serial dilutions Figure 6.16

19. Plate Count After incubation, count colonies on plates that have 25-250 colonies (CFUs) Figure 6.15

20. Direct Measurements of Microbial Growth Filtration Figure 6.17

21. Direct Measurements of Microbial Growth Multiple tube MPN test. Count positive tubes and compare to statistical MPN table. Figure 6.18b

22. Direct Measurements of Microbial Growth Direct microscopic count

23. Direct Measurements of Microbial Growth Figure 6.19, steps 1, 3

24. Estimating Bacterial Numbers by Indirect Methods Turbidity Figure 6.20

25. Measuring Microbial Growth Direct methods Plate counts Filtration MPN Direct microscopic count Dry weight Indirect methods Turbidity Metabolic activity Dry weight