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Microbial Growth

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

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The Requirements for Growth

• Physical requirements
– Temperature
– pH
– Osmotic pressure
• Chemical requirements
– Carbon
– Nitrogen, sulfur, and phosphorous
– Trace elements
– Oxygen
– Organic growth factor

The Requirements for Growth:

Physical Requirements

• Temperature
– Minimum growth temperature
– Optimum growth temperature
– Maximum growth temperature

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Temperature

Figure 6.1

Growth Temperature

Figure 6.2

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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

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

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The Requirements for Growth:

Physical Requirements

Figure 6.4

The Requirements for Growth:

Chemical Requirements

• Carbon
– Structural organic molecules, energy source
– Chemoheterotrophs use organic carbon sources
– Autotrophs use CO2

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• 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

• Trace Elements
– Inorganic elements required in small amounts
– Usually as enzyme cofactors

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• Oxygen (O2)
Obligate Faultative Obligate Aerotolerant
Microaerophiles
aerobes anaerobes anaerobes anaerobes

Biofilms

• Microbial
communities
• Form slime or
hydrogels
– Bacteria attracted
by chemicals via
quorum sensing

Figure 6.5

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The Requirements for Growth:

Chemical Requirements

• Organic Growth Factors

– Organic compounds obtained from the
environment
– Vitamins, amino acids, purines, pyrimidines

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

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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

Culture Media

• Chemically Defined Media: Exact chemical

composition is known
• Complex Media: Extracts and digests of yeasts,
meat, or plants
– Nutrient broth
– Nutrient agar

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Culture Media

Table 6.2 & 6.4

Anaerobic Culture Methods

• Reducing media
– Contain chemicals (thioglycollate or oxyrase) that
combine O2
– Heated to drive off O2

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An Anaerobic Chamber

Figure 6.7

Anaerobic Culture Methods

• Anaerobic jar

Figure 6.5

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Capnophiles require high CO2

• Candle jar

• CO2-packet

Figure 6.7

Biosafety Levels

• 1: No special precautions
• 2: Lab coat, gloves, eye protection
• 3: Biosafety cabinets to prevent airborne
transmission
• 4: Sealed, negative pressure
– Exhaust air is filtered twice

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Biosafety Level 4 (BSL-4) Laboratory

Figure 6.8

Selective Media

• To chemically (or
physically) suppress
unwanted microbes
and encourage
desired microbes.

Figure 6.9b, c

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Selective Media
Mannitol Salt Agar : selective for MSA
halophiles with 7% salt (osmotic
challenge) and differential for mannitol
fermenters; good for skin bacterial
cultures.

EMB Agar: kills Gram positives with eosin EMB

and methylene blue, selective for Gram
negatives. Differential for lactose
fermenters. Good for growing enterics.

McConkey Agar: supresses Gram MA

positives with crystal violet and bile salts

Differential Media
• Make it easy to distinguish colonies of different
microbes.
• Distinguish between different species based on
a metabolic ability.

Mannitol salt agar

Blood agar (sheep’s blood) reveals if hemolytic contains the pH sensitive
dye phenol red (yellow
when acidic)

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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
– Only phenol-metabolizing bacteria will be growing

• 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

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

Binary Fission

Figure 6.11

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Figure 6.12b

If 100 cells growing for 5 hours produced

1,720,320 cells:

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Figure 6.13

Phases of Growth

ANIMATION Bacterial Growth Curve

Figure 6.15

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Direct Measurements of
Microbial Growth
• Plate Counts: Perform serial dilutions of a
sample

Figure 6.15, top portion

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 or 30-300 colonies (CFUs)

Figure 6.15

Direct Measurements of
Microbial Growth
• Filtration

Figure 6.17a, b

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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

Direct Measurements of
Microbial Growth
• Direct Microscopic Count

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Direct Measurements of Microbial

Growth

Figure 6.19

Estimating Bacterial Numbers by

Indirect Methods

• Turbidity

Figure 620

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Estimating Bacterial Numbers by

Indirect methods

• Metabolic activity
• Dry weight

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