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Chapter 6, part A

Chapter 6, part A. Microbial Growth. Microbial Growth. Microbial growth = increase in number of cells, not cell size. The Requirements for Growth: Physical Requirements. Temperature Minimum growth temperature Optimum growth temperature Maximum growth temperature. Temperature. Figure 6.1.

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Chapter 6, part A

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  1. Chapter 6, part A Microbial Growth

  2. Microbial Growth • Microbial growth = increase in number of cells, not cell size

  3. The Requirements for Growth: Physical Requirements • Temperature • Minimum growth temperature • Optimum growth temperature • Maximum growth temperature

  4. Temperature Figure 6.1

  5. Psychrotrophs • Grow between 0°C and 20-30°C • Cause food spoilage

  6. Psychrotrophs Figure 6.2

  7. The Requirements for Growth: Physical Requirements • pH • Most bacteria grow between pH 6.5 and 7.5 • Molds and yeasts grow between pH 5 and 6 • Acidophiles grow in acidic environments

  8. The Requirements for Growth: Physical Requirements • Osmotic Pressure • Hypertonic environments, increase salt or sugar, cause plasmolysis • Extreme or obligate halophiles require high osmotic pressure • Facultative halophiles tolerate high osmotic pressure

  9. The Requirements for Growth: Physical Requirements Figure 6.4

  10. The Requirements for Growth: Chemical Requirements • Carbon • Structural organic molecules, energy source • Chemoheterotrophs use organic carbon sources • Autotrophs use CO2

  11. The Requirements for Growth: Chemical Requirements • Nitrogen • In amino acids, proteins • Most bacteria decompose proteins • Some bacteria use NH4+ or NO3 • A few bacteria use N2 in nitrogen fixation • Sulfur • In amino acids, thiamine, biotin • Most bacteria decompose proteins • Some bacteria use SO42 or H2S • Phosphorus • In DNA, RNA, ATP, and membranes • PO43is a source of phosphorus

  12. The Requirements for Growth: Chemical Requirements • Trace Elements • Inorganic elements required in small amounts • Usually as enzyme cofactors

  13. The Requirements for Growth: Chemical Requirements • Oxygen (O2)

  14. Toxic Forms of Oxygen • Singlet oxygen: O2 boosted to a higher-energy state • Superoxide free radicals: O2 • Peroxide anion: O22 • Hydroxyl radical (OH)

  15. The Requirements for Growth: Chemical Requirements • Organic Growth Factors • Organic compounds obtained from the environment • Vitamins, amino acids, purines, pyrimidines

  16. Culture Media • Culture Medium: Nutrients prepared for microbial growth • Sterile: No living microbes • Inoculum: Introduction of microbes into medium • Culture: Microbes growing in/on culture medium

  17. Agar • Complex polysaccharide • Used as solidifying agent for culture media in Petri plates, slants, and deeps • Generally not metabolized by microbes • Liquefies at 100°C • Solidifies ~40°C

  18. Culture Media • Chemically Defined Media: Exact chemical composition is known • Complex Media: Extracts and digests of yeasts, meat, or plants • Nutrient broth • Nutrient agar

  19. Culture Media Table 6.2 & 6.4

  20. Anaerobic Culture Methods • Reducing media • Contain chemicals (thioglycollate or oxyrase) that combine O2 • Heated to drive off O2

  21. Anaerobic Culture Methods • Anaerobic jar Figure 6.5

  22. Anaerobic Culture Methods • Anaerobic chamber Figure 6.6

  23. Capnophiles require high CO2 • Candle jar • CO2-packet Figure 6.7

  24. Selective Media • Suppress unwanted microbes and encourage desired microbes. Figure 6.9b, c

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

  26. 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

  27. 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)

  28. Streak Plate Figure 6.10a, b

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

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

  31. Binary Fission Figure 6.11

  32. Figure 6.12b

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

  34. Figure 6.13

  35. Figure 6.14

  36. Direct Measurements of Microbial Growth • Plate Counts: Perform serial dilutions of a sample Figure 6.15, top portion

  37. Plate Count • Inoculate Petri plates from serial dilutions Figure 6.16

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

  39. Direct Measurements of Microbial Growth • Filtration Figure 6.17a, b

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

  41. Direct Measurements of Microbial Growth • Direct Microscopic Count

  42. Direct Measurements of Microbial Growth Figure 6.19

  43. Estimating Bacterial Numbers by Indirect Methods • Turbidity Figure 620

  44. Estimating Bacterial Numbers by Indirect methods • Metabolic activity • Dry weight

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