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Chapter 6
Microbial
Nutrition and
Growth
Growth Requirements
Microbial
growth
 Increase in a population of microbes
Result
colony
of microbial growth is discrete
 An aggregation of cells arising from single
parent cell
Reproduction
© 2012 Pearson Education Inc.
results in growth
Growth Requirements
Organisms
use a variety of nutrients for
their energy needs and to build organic
molecules and cellular structures
Most common nutrients contain
necessary elements such as carbon,
oxygen, nitrogen, and hydrogen
Microbes obtain nutrients from variety
of sources
© 2012 Pearson Education Inc.
Growth Requirements
Nutrients:
Chemical and Energy
Requirements
 Sources of carbon, energy, and electrons
○ Two groups of organisms based on source of
carbon
 Autotrophs
 Heterotrophs
○ Two groups of organisms based on source of
energy
 Chemotrophs
 Phototrophs
© 2012 Pearson Education Inc.
Growth Requirements
 Nutrients:
Chemical and Energy Requirements
 Oxygen requirements
○ Oxygen is essential for obligate aerobes
○ Oxygen is deadly for obligate anaerobes
 How can this be true?
- O2 and oxygen in compounds are not toxic
- However, toxic forms of oxygen are highly reactive and
excellent oxidizing agents (same reason why oxygen is
excellent electron acceptor in electron transport chain)
- Resulting oxidation causes irreparable damage to cells
- Obligate anaerobes don’t have enzymes to deal with
these toxic forms of oxygen
© 2012 Pearson Education Inc.
Growth Requirements
 Nutrients:
Chemical and Energy Requirements
 Oxygen requirements
○ Four toxic forms of oxygen
 Singlet oxygen
- Can be formed during aerobic respiration, some
organisms have enzymes to deal with this
 Superoxide radicals
 Peroxide anion
 Hydroxyl radical
© 2012 Pearson Education Inc.
Growth Requirements

Nutrients: Chemical and Energy Requirements
 Oxygen requirements
○ Aerobes
○ Anaerobes
○ Facultative anaerobes
 Can grow without oxygen but growth is enhanced with it
○ Aerotolerant anaerobes
 Are not bothered by presence of oxygen
○ Microaerophiles
 Need lower % oxygen
© 2012 Pearson Education Inc.
Growth Requirements
Nutrients:
Chemical and Energy
Requirements
 Nitrogen requirements
○ Anabolism often ceases because of insufficient
nitrogen
○ Nitrogen acquired from organic and inorganic
nutrients
○ All cells recycle nitrogen from amino acids and
nucleotides
○ Nitrogen fixation by certain bacteria is essential
to life on Earth
© 2012 Pearson Education Inc.
Growth Requirements
Nutrients:
Chemical and Energy
Requirements
 Other chemical requirements
○ Phosphorus
○ Sulfur
○ Trace elements – vitamins, minerals, etc
 Required only in small amounts
○ Growth factors
 Necessary organic chemicals that cannot be
synthesized by certain organisms
© 2012 Pearson Education Inc.
Growth Requirements
Physical
Requirements
 Temperature
○ Effect of temperature on
proteins
○ Effect of temperature on
membranes of cells and
organelles
 If too low, membranes become
rigid and fragile
 If too high, membranes become
too fluid
© 2012 Pearson Education Inc.
Growth Requirements
Physical Requirements
 pH
○ Organisms are sensitive to changes in acidity
 H+ and OH– interfere with H bonding
○ Neutrophiles grow best in a narrow range
around neutral pH
○ Acidophiles grow best in acidic habitats
○ Alkalinophiles live in alkaline soils and water
© 2012 Pearson Education Inc.
Growth Requirements
 Physical
Requirements
 Physical effects of water
○ Microbes require water to dissolve enzymes and nutrients
○ Water is important reactant in many metabolic reactions
○ Most cells die in absence of water
 Some have cell walls that retain water
 Endospores and cysts cease most metabolic activity
○ Two physical effects of water
 Osmotic pressure
 Hydrostatic pressure
© 2012 Pearson Education Inc.
Growth Requirements

Physical Requirements
 Physical effects of water
○ Osmotic pressure
 Pressure exerted on a semipermeable membrane by a
solution containing solutes that cannot freely cross
membrane
 Hypotonic solutions have lower solute concentrations
 Hypertonic solutions have greater solute
concentrations
 Restricts organisms to certain environments
- Obligate and facultative halophiles
© 2012 Pearson Education Inc.
Growth Requirements

Physical Requirements
 Physical effects of water
○ Hydrostatic pressure
 Water exerts pressure in proportion to its
depth
 Barophiles live under extreme pressure
- Their membranes and enzymes depend on
pressure to maintain their shape
© 2012 Pearson Education Inc.
Growth Requirements
Associations
and Biofilms
 Organisms live in association with different
species
○ Antagonistic relationships
- One organism is harmed or killed
○ Synergistic relationships
- Organisms are benefited by relationship but can live fine
on their own
○ Symbiotic relationships
- Organisms are interdependent
© 2012 Pearson Education Inc.
Growth Requirements
Associations and Biofilms
 Biofilms
○ Complex relationships among numerous
microorganisms
○ Develop an extracellular matrix




Adheres cells to one another
Allows attachment to a substrate
Sequesters nutrients
May protect individuals in the biofilm
○ Form on surfaces often as a result of quorum
sensing
○ Many microorganisms more harmful as part of a
biofilm
© 2012 Pearson Education Inc.
Culturing Microorganisms
Inoculum
medium
(specimen) introduced into
 Environmental specimens
 Clinical specimens
 Stored specimens
Culture
 Act of cultivating microorganisms or the
microorganisms that are cultivated
© 2012 Pearson Education Inc.
Culturing Microorganisms
Figure 6.10 Pour plate method of isolation-overview

Obtaining Pure Cultures
 Cultures composed of cells arising from a single progenitor
○ Progenitor is termed a CFU (colony forming unit)
 COLONY – group of bacteria arising from a single bacteria (in other words – a
bunch of clones)
 Aseptic technique prevents contamination of sterile substances or
objects
 Two common isolation techniques:
○ Streak plates
○ Pour plates
© 2012 Pearson Education Inc.
Culturing Microorganisms
 Culture Media
 Majority of prokaryotes have not been grown in culture
medium
 Six types of general culture media
○ Defined media
- Synthetic, exact chemical composition is known
○ Complex media
- Contains nutrients that are released by partial digestion of
yeast, beef, soy
- Used to grow organisms whose exact nutritional needs are not
known
- TSA, MacConkey, nutrient broth, blood is sometimes added
○ Selective media
- Either favors growth of particular organism or inhibits
○ Differential media
- Differences in change in media or colony color can be seen
○ Anaerobic media
○ Transport media
- Depends on sample and suspected organism
© 2012 Pearson Education Inc.
Culturing Microorganisms
Special Culture Techniques
 Techniques developed for culturing
microorganisms
○ Animal and cell culture
○ Low-oxygen culture
 CO2 incubators
○ Enrichment culture
 Uses selective media to enhance #’s of a particular organism in
mixed sample
© 2012 Pearson Education Inc.
Culturing Microorganisms
Preserving
Cultures
 Refrigeration
○ Stores for short periods of time
 Deep-freezing
○ Stores for years
 Lyophilization - removing water from
frozen culture
○ Stores for decades
© 2012 Pearson Education Inc.
Growth of Microbial Populations
Generation
Time
 Time required for a bacterial cell to grow
and divide
 Dependent on chemical and physical
conditions
© 2012 Pearson Education Inc.
Number of live cells (log)
Stationary phase
Log
(exponential)
phase
Lag phase
Time
Death
(decline)
phase
Growth of Microbial Populations
Measuring
Microbial Reproduction
 Direct methods
○ Serial dilution and viable plate counts
○ Membrane filtration
○ Most probable number
○ Microscopic counts
○ Electronic counters
© 2012 Pearson Education Inc.
Growth of Microbial Populations
Measuring
Microbial Growth
 Indirect methods
○ Metabolic activity under standard
conditions
○ Dry weight
 Filamentous organisms difficult to count; dry and
weigh
○ Turbidity
© 2012 Pearson Education Inc.
Growth of Microbial Populations
Measuring
Microbial Reproduction
 Genetic methods
○ Isolate DNA sequences of unculturable
prokaryotes
 Majority of bacteria and archaea cannot be grown
in lab
 Used to estimate the number of these
microbes
© 2012 Pearson Education Inc.