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Microbiology: A Systems

Approach, 2nd ed.


Chapter 7: Microbial
Nutrition, Ecology, and
Growth
7.1 Microbial Nutrition
• Nutrition: a process by which chemical substances
(nutrients) are acquired from the environment and
used in cellular activities
• All living things require a source of elements such as
C, H, O, P, K, N, S, Ca, Fe, Na, Cl, Mg- but the relative
amounts vary depending on the microbe
• Essential Nutrient: any substances that must be
provided to an organism
– Macronutrients: Required in relatively large
quantities, play principal roles in cell structure and
metabolism (ex. C, H, O)
– Micronutrients: aka trace elements, present in smaller
amounts and involved in enzyme function and
maintenance of protein structure (ex. Mn, Zn, Ni)
• Nutrients are processed and transformed into the
chemicals of the cell after absorption
• Can also categorize nutrients according to C content
– Inorganic nutrients: A combination of atoms other than
C and H
– Organic nutrients: Contain C and H, usually the products
Chemical Analysis of Microbial
Cytoplasm
Sources of Essential
Nutrients
• Carbon sources
• Nitrogen sources
• Oxygen sources
• Hydrogen sources
• Phosphorus sources
• Sulfur sources
• Others
Carbon Sources
• The majority of C compounds
involved in normal structure and
metabolism of all cells are organic
• Heterotroph: Must obtain C in
organic form (nutritionally dependent
on other living things)
• Autotroph: Uses inorganic CO2 as
its carbon source (not nutritionally
dependent on other living things)
Nitrogen Sources
• Main reservoir- N2
• Primary nitrogen source for
heterotrophs- proteins, DNA, RNA
• Some bacteria and algae utilize
inorganic nitrogenous nutrients
• Small number can transform N2 into
usable compounds through nitrogen
fixation
• Regardless of the initial form, must be
converted to NH3 (the only form that
can be directly combined with C to
synthesize amino acids and other
compounds)
Oxygen Sources
• O is a major component of organic
compounds
• Also a common component of
inorganic salts
• O2 makes up 20% of the atmosphere
Hydrogen Sources
• H is a major element in all organic
and several inorganic compounds
• Performs overlapping roles in the
biochemistry of cells:
– Maintaining pH
– Forming hydrogen bonds between
molecules
– Serving as the source of free energy in
oxidation-reduction reactions of
respiration
Phosphorus (Phosphate)
Sources
• Main inorganic source of phosphorus
is phosphate (PO43-)
– Derived from phosphoric acid
– Found in rocks and oceanic mineral
deposits
• Key component in nucleic acids
• Also found in ATP
• Phospholipids in cell membranes and
coenzymes
Sulfur Sources
• Widely distributed throughout the
environment in mineral form
• Essential component of some
vitamins
• Amino acids- methionine and
cysteine
Other Nutrients Important in
microbial Metabolism
• Potassium- protein synthesis and membrane
function
• Sodium- certain types of cell transport
• Calcium- stabilizer of cell walls and
endospores
• Magnesium- component of chlorophyll and
stabilizer of membranes and ribosomes
• Iron- important component of cytochrome
proteins
• Zinc- essential regulatory element for
eukaryotic genetics, and binding factors for
enzymes
• Cooper, cobalt, nickel, molybdenum,
manganese, silicon, iodine, and boron-
Growth Factors: Essential
Organic Nutrients
• Growth factor: An organic
compound such as an amino acid,
nitrogenous base, or vitamin that
cannot be synthesized by an
organism and must be provided as a
nutrient
• For example, many cells cannot
synthesize all 20 amino acids so they
must obtain them from food
(essential amino acids)
How Microbes Feed:
Nutritional Types
Main Determinants of
Nutritional Type
• Sources of carbon and energy
• Phototrophs- Microbes that
photosynthesize
• Chemotrophs- Microbes that gain
energy from chemical compounds
Autotrophs and Their Energy
Sources
• Photoautotrophs
– Photosynthetic
– Form the basis for most food webs
• Chemoautotrophs
– Chemoorganic autotrophs- use organic
compounds for energy and inorganic
compounds as a carbon source
– Lithoautotrophs- rely totally on inorganic
minerals
– Methanogens- produce methane from
hydrogen gas and carbon dioxide
• Archae
• Some live in extreme habitats
Figure 7.1
Heterotrophs and Their Energy
Sources
– Majority are chemoheterotrophs that
derive both carbon and energy from
organic compounds
• Saprobes
– Free-living microorganisms
– Feed primarily on organic detritus from dead
organisms
– Decomposers of plant litter, animal matter, and
dead microbes
– Most have rigid cell wall, so they release
enzymes to the extracellular environment and
digest food particles into smaller molecules
» Obligate saprobes- exist strictly on dead organic
matter in soil and water
» Facultative parasite- when a saprobe infects a host,
usually when the host is compromised (opportunistic
pathogen)
Figure 7.2
Other Chemoheterotrophs
• Parasites
– Derive nutrients from the cells or tissues
of a host
– Also called pathogens because they
cause damage to tissues or even death
– Ectoparasites- live on the body
– Endoparasites- live in organs and tissues
– Intracellular parasites- live within cells
– Obligate parasites- unable to grow
outside of a living host
Transport Mechanisms for
Nutrient Absorption
• Cells must take nutrients in and
transport waste out
• Transport occurs across the cell
membrane, even in organisms with
cell walls
The Movement of Water:
Osmosis
• Osmosis: Diffusion of water through a
selectively permeable membrane
• The membrane is selectively
permeable- having passageways that
allow free diffusion of water but can
block certain other dissolved molecules
• When the membrane is between
solutions of differing concentrations and
the solute is not diffusible, water will
diffuse at a fast rate from the side that
has more water to the side that has less
water
Figure 7.3
Osmotic Relationships
• The osmotic relationship between cells and their
environment is determined by the relative concentrations
of the solutions on either side of the cell membrane
• Isotonic: The environment is equal in solute concentration
to the cell’s internal environment
– No net change in cell volume
– Generally the most stable environment for cells
• Hypotonic: The solute concentration of the external
environment is lower than that of the cell’s internal
environment
– Net direction of osmosis is from the hypotonic solution into the
cell
– Cells without cell walls swell and can burst
• Hypertonic: The environment has a higher solute
concentration than the cytoplasm
– Will force water to diffuse out of a cell
– Said to have high osmotic pressure
Figure 7.4
Adaptations to Osmotic
Variations in the Environment
• Example: fresh pond water-
hypotonic conditions
– Bacteria- cell wall protects them from
bursting
– Amoeba- a water (or contractile)
vacuole that moves excess water out of
the cell
• Example: high-salt environment-
hypertonic conditions
– Halobacteria living in the Great Salt
The Movement of Molecules:
Diffusion and Transport
• Diffusion: When atoms or molecules move in a
gradient from an area of higher density or
concentration to an area of lower density or
concentration
– Random thermal movement of molecules will
eventually distribute the molecules from an area
of higher concentration to an area of lower
concentration
– Evenly distributes the molecules
– Diffusion of molecules across the cell membrane
is largely determined by the concentration
gradient and permeability of the substance
– Simple or passive diffusion is limited to small
nonpolar molecules or lipid soluble molecules
Figure 7.5
Facilitated Diffusion
• Utilizes a carrier protein that binds a
specific substance, changes the
conformation of the carrier protein, and the
substance is moved across the membrane
• Once the substance is transported, the
carrier protein resumes its original shape
• Carrier proteins exhibit specificity
• Saturation: The rate of a substance is
limited by the number of binding sites on
the transport proteins
• Competition: When two molecules of
similar shape can bind to the same binding
site on a carrier protein
Figure 7.6
Active Transport
• Nutrients are transported against the diffusion
gradient or in the same direction as the natural
gradient but at a rate faster than by diffusion
alone
• Requires the presence of specific membrane
proteins (permeases and pumps)
• Requires the expenditure of energy
• Items that require active transport:
monosaccharides, amino acids, organic acids,
phosphates, and metal ions
• Specialized pumps- an important type of active
transport
• Group translocation: couples the transport of
a nutrient with its conversion to a substance
that is immediately useful inside the cell
Figure 7.7
Endocytosis: Eating and
Drinking by Cells
• A form of active transport
• Transporting large molecules, particles,
lipids, or other cells
• Occurs in some eukaryotic cells
• The cell encloses the substance in its cell
membrane, simultaneously forming a
vacuole and engulfing it
• Phagocytosis- amoebas and certain white
blood cells; ingesting whole cells or large
solid matter
• Pinocytosis- Transport of liquids such as
oils or molecules in solution
7.2 Environmental Factors
that Influence Microbes
• Temperature Adaptations
– Microbial cells cannot control their temperature, so they
assume the ambient temperature of their natural habitat
– The range of temperatures for the growth of a given
microbial species can be expressed as three cardinal
temperatures:
• Minimum temperature: the lowest temperature that
permits a microbe’s continued growth and metabolism
• Maximum temperature: The highest temperature at which
growth and metabolism can proceed
• Optimum temperature: A small range, intermediate
between the minimum and maximum, which promotes the fast
rate of growth and metabolism
– Some microbes have a narrow cardinal range while
others have a broad one
– Another way to express temperature adaptation- to
describe whether an organism grows optimally in a cold,
moderate, or hot temperature range
Psychophile
• A microorganism that has an
optimum temperature below 15°C
and is capable of growth at 0°C.
• True psychrophiles are obligate with
respect to cold and cannot grow
above 20°C.
• Psychrotrophs or facultative
psychrophiles- grow slowly in cold
but have an optimum temperature
above 20°C.
Figure 7.8
Figure 7.9
Mesophile
• An organism that grows at
intermediate temperatures
• Optimum growth temperature of
most: 20°C to 40°C
• Temperate, subtropical, and tropical
regions
• Most human pathogens have optima
between 30°C and 40°C
Thermophile
• A microbe that grows optimally at
temperatures greater than 45°C
• Vary in heat requirements
• General range of growth of 45°C to
80°C
• Hyperthermophiles- grow between
80°C and 120°C
Gas Requirements
• Atmospheric gases that most influence
microbial growth- O2 and CO2
• Oxygen gas has the greatest impact on
microbial growth
• As oxygen enters into cellular reactions,
it is transformed into several toxic
products
– Most cells have developed enzymes that go
about scavenging and neutralizing these
chemicals
• Superoxide dismutase
• Catalase
– Essential for aerobic organisms
Several General Categories of
Oxygen Requirements
• Aerobe: can use gaseous oxygen in its metabolism
and possesses the enzymes needed to process toxic
oxygen products
• Obligate aerobe: cannot grow without oxygen
• Facultative anaerobe: an aerobe that does not
require oxygen for its metabolism and is capable of
growth in the absence of it
• Microaerophile: does not grow at normal
atmospheric concentrations of oxygen but requires a
small amount of it in metabolism
• Anaerobe: lacks the metabolic enzyme systems for
using oxygen in respiration
• Strict, or obligate, anaerobes: also lack the
enzymes for processing toxic oxygen and cannot
tolerate any free oxygen in the immediate
environment and will die if exposed to it.
• Aerotolerant anaerobes: do not utilize oxygen but
can survive and grow to a limited extent in its
Figure 7.10
Figure 7.11
Carbon Dioxide
• All microbes require some carbon
dioxide in their metabolism
• Capnophiles grow best at a higher
CO2 tension than is normally present
in the atmosphere
Effects of pH
• Majority of organisms live or grow in
habitats between pH 6 and 8
• Obligate acidophiles
– Euglena mutabilis- alga that grows
between 0 and 1.0 pH
– Thermoplasma- archae that lives in hot
coal piles at a pH of 1 to 2, and would
lyse if exposed to pH 7
Osmotic Pressure
• Most microbes live either under
hypotonic or isotonic conditions
• Osmophiles- live in habitats with a
high solute concentration
• Halophiles- prefer high
concentrations of salt
• Obligate halophiles- grow optimally
in solutions of 25% NaCl but require
at least 9% NaCl for growth
Miscellaneous Environmental
Factors
• Nonphotosynthetic microbes tend to be
damaged by the toxic oxygen products
produced by contact with light
– Some produce yellow carotenoid pigments to
protect against the damaging effects of light by
absorbing and dismantling toxic oxygen
• Other types of radiation that can damage
microbes are ultraviolet and ionizing rays
• Barophiles: deep-sea microbes that exist
under hydrostatic pressures ranging from a few
times to over 1,000 times the pressure of the
atmosphere
• All cells require water- only dormant,
dehydrated cell stages tolerate extreme drying
Ecological Associations Among
Microorganisms
• Most microbes live in shared habitats
• Interactions can have beneficial,
harmful, or no particular effects on
the organisms involved
• They can be obligatory or
nonobligatory to the members
• They often involve nutritional
interactions
Symbiosis
• A general term used to denote a situation in
which two organisms live together in a close
partnership
– Members are termed symbionts
– Three main types of symbionts
• Mutualism: when organisms live in an obligatory but
mutually beneficial relationship
• Commensalism: the member called the commensal
receives benefits, while its coinhabitant is neither
harmed nor benefited
– Satellitism: when one member provides nutritional or
protective factors needed by the other
• Parasitism: a relationship in which the host
organism provides the parasitic microbe with
nutrients and a habitat
Figure 7.12
Nonsymbiotic Relationship
• Organisms are free-living and
relationships are not required for
survival
– Synergism: an interrelationship
between two or more free-living
organisms that benefits them but is not
necessary for their survival
– Antagonism: an association between
free-living species that arrises when
members of a community compete
• One microbe secretes chemical substances
into the surrounding environment that
inhibit or destroy another microbe in the
Interrelationships Between
microbes and Humans
• Normal microbiotia: microbes that
normally live on the skin, in the
alimentary tract, and in other sites in
humans
• Can be commensal, parasitic, and
synergistic relationships
7.3 The Study of Microbial
Growth
• Growth takes place on two levels
– Cell synthesizes new cell components
and increases in size
– The numer of cells in the population
increases
• The Basis of Population Growth:
Binary Fission
Figure 7.13
Figure 7.14
The Rate of Population
Growth
– Generation or doubling time: The time
required for a complete fission cycle
– Each new fission cycle or generation
increases the population by a factor of 2
– As long as the environment is favorable, the
doubling effect continues at a constant rate
– The length of the generation time- a
measure of the growth rate of an organism
• Average generation time- 30 to 60 minutes
under optimum conditions
• Can be as short as 10 to 12 minutes
– This growth pattern is termed exponential
Graphing Bacterial Growth
• The data from growing bacterial
populations are graphed by plotting the
number of cells as a function of time
– If plotted logarithmically- a straight line
– If plotted arithmetically- a constantly
curved slope
• To calculate thesize of a population
over time: Nf = (Ni)2n
– Nf is the total number of cells in the
population at some point in the growth
phase
– Ni is the starting number
– N denotes the generation number
The Population Growth
Curve
• A population of bacteria does not maintain
its potential growth rate and double
endlessly
• A population displays a predictable pattern
called a growth curve
• The method to observe the population
growth pattern:
– Place a tiny number of cells in a sterile liquid
medium
– Incubate this culture over a period of several
hours
– Sampling the browth at regulat intervals during
incubation
Stages in the Normal
Growth Curve
• Data from an entire growth period
typically produce a curve with a
series of phases
• Lag Phase
• Exponential Growth Phase
• Stationary Growth Phase
• Death Phase
Lag Phase
• Relatively “flat” period
• Newly inoculated cells require a
period of adjustment, enlargement,
and synthesis
• The cells are not yet multiplying at
their maximum rate
• The population of cells is so sparse
that the sampling misses them
• Length of lag period varies from one
population to another
Exponential Growth
(Logarithmic or log) Phase
• When the growth curve increases
geometrically
• Cells reach the maximum rate of cell
division
• Will continue as long as cells have
adequate nutrients and the
environment is favorable
Stationary Growth Phase
• The population enters a survival
mode in which cells stop growing or
grow slowly
– The rate of cell inhibition or death
balances out the rate of multiplication
– Depleted nutrients and oxygen
– Excretion of organic acids and other
biochemical pollutants into the growth
medium
Death Phase
• The curve dips downward
• Cells begin to die at an exponential
rate
Figure 7.15
Potential Importance of the
Growth Curve
• Implications in microbial control,
infection, food microbiology, and
culture technology
• Growth patterns in microorganisms
can account for the stages of
infection
• Understanding the stages of cell
growth is crucial for working with
cultures
• In some applications, closed batch
culturing is inefficient, and instead,
Other Methods of Analyzing
Population Growth
– Turbidometry- a tube of clear nutrient
solution becomes turbid as microbes
grow in it
Figure 7.16
Enumeration of Bacteria
• Direct or total cell count- counting
the number of cells in a sample
microscopically
– Uses a special microscope slide
(cytometer)
– Used to estimate the total number of
cells in a larger sample
Figure 7.17
Automated Counting
• Coulter counter- electronically scans
a culture as it passes through a tiny
pipette
• Flow cytometer also measures cell
size and differentiates between live
and dead cells
• Real-time PCR allows scientists to
quantify bacteria and other
microorganisms that are present in
environmental or tissue samples
Figure 7.18