Growth may be defined as an increase in cellular constituents.

It leads to a rise in
cell number when microorganisms reproduce by processes like budding or binary
fission.

Growth also results when cells simply become longer or larger. If the
microorganism is coenocytic—that is, a multinucleate organism in which nuclear
divisions are not accompanied by cell divisions— growth results in an increase in
cell size but not cell number.

When studying growth, microbiologists normally follow changes in the total

population number.

Population growth is studied by analyzing the growth curve of a microbial culture

The growth of microorganisms reproducing by binary fission can be plotted as the

logarithm of the number of viable cells versus the incubation time. The resulting
curve has four distinct phases.
This phase may be quite long if the inoculum is from an old culture or one that has
been refrigerated.

Inoculation of a culture into a chemically different medium also results in a longer

lag phase.

On the other hand, when a young, vigorously growing exponential phase culture is
transferred to fresh medium of the same composition, the lag phase will be.

Exponential Phase

During the exponential or log phase, microorganisms are growing and dividing at the
maximal rate possible given their genetic potential, the nature of the medium, and
the conditions under which they are growing.

Their rate of growth is constant during the exponential phase; that is, the
microorganisms are dividing and doubling in number at regular intervals. Because
each individual divides at a slightly different moment, the growth curve rises
smoothly rather than in discrete jumps.
The population is most uniform in terms of chemical and physiological properties during
this phase; therefore exponential phase cultures are usually used in biochemical and
physiological studies.

Exponential growth is balanced growth. That is, all cellular constituents are
manufactured at constant rates relative to each other.

If nutrient levels or other environmental conditions change, unbalanced growth results.

This is growth during which the rates of synthesis of cell components vary relative to
one another until a new balanced state is reached. This response is readily observed in a
shift-up experiment in which bacteria are transferred from a nutritionally poor medium
to a richer one.

Unbalanced growth also results when a bacterial population is shifted down from a rich
medium to a poor one. The organisms may previously have been able to obtain many
cell components directly from the medium.

When shifted to a nutritionally inadequate medium, they need time to make the
enzymes required for the biosynthesis of unavailable nutrients. Consequently cell
division and DNA replication continue after the shift-down, but net protein and RNA
synthesis slow.
Stationary Phase

Eventually population growth ceases and the growth curve becomes horizontal

This stationary phase usually is attained by bacteria at a population level of around

109 cells per ml.

Other microorganisms normally do not reach such high population densities,

protozoan and algal cultures often having maximum concentrations of about 106 cells
per ml.

Final population size depends on nutrient availability and other factors, as well as the
type of microorganism being culture.

In the stationary phase the total number of viable microorganisms remains constant.
This may result from a balance between cell division and cell death, or the population
may simply cease to divide though remaining metabolically active.
Reasons

Nutrient limitation, if an essential nutrient is severely depleted, population growth

will slow.

Aerobic organisms often are limited by O2 availability. Oxygen is not very soluble and
may be depleted so quickly that only the surface of a culture will have an O2
concentration adequate for growth.

The cells beneath the surface will not be able to grow unless the culture is shaken or
aerated in another way.

Population growth also may cease due to the accumulation of toxic waste products.
This factor seems to limit the growth of many anaerobic cultures.

For example, streptococci can produce so much lactic acid and other organic acids
from sugar fermentation that their medium becomes acidic.
Death Phase

Detrimental environmental changes like nutrient deprivation and the buildup of toxic
wastes lead to the decline in the number of viable cells characteristic of the death phase.
Influence of Environmental Factors on Growth
 The growth of microorganisms also is greatly affected by the chemical and
physical nature of their surroundings.

 An understanding of environmental influences aids in the control of microbial

growth and the study of the ecological distribution of microorganisms.

 The most important environmental factors affect microbial growth are solutes
and water activity, pH, temperature, oxygen level, pressure, and radiation.

Solutes and Water Activity

 A selectively permeable plasma membrane separates microorganisms from

their environment, they can be affected by changes in the osmotic concentration
of their surroundings.

 A microorganism is placed in a hypotonic solution (one with a lower osmotic

concentration), water will enter the cell and cause it to burst unless something is
done to prevent the influx.
 The osmotic concentration of the cytoplasm can be reduced by use of inclusion
bodies .

 Prokaryotes also can contain pressure-sensitive channels that open to allow

solute escape when the osmolarity of the environment becomes much lower than
that of the cytoplasm.

 When microorganisms with rigid cell walls are placed in a hypertonic

environment, water leaves and the plasma membrane shrinks away from the wall,
a process known as plasmolysis. This dehydrates the cell and may damage the
plasma membrane; the cell usually becomes metabolically inactive and ceases to
grow.

 Many microorganisms keep the osmotic concentration of their protoplasm

somewhat above that of the habitat by the use of compatible solutes.

 Compatible solutes are solutes that are compatible with metabolism and growth
when at high intracellular concentrations.

 Most prokaryotes increase their internal osmotic concentration in a hypertonic

environment through the synthesis or uptake of choline, betaine, proline,
glutamic acid, and other amino acids elevated levels of potassium ions are also
involved to some extent.
Ex: Algae and fungi employ sucrose and polyols.

Halobacterium salinarium raise their osmotic concentration with potassium ions.

 Protozoa do not have a cell wall, they must use contractile vacuoles to eliminate
excess water when living in hypotonic environments.

 Microorganisms differ greatly in their ability to adapt to habitats with low water
activity.

 A microorganism must expend extra effort to grow in a habitat with a low aw

value because it must maintain a high internal solute concentration to retain
water.

Ex: Staphylococcus aureus can be cultured in media containing any sodium chloride
concentration up to about 3 M.

yeast Saccharomyces rouxii will grow in sugar solutions with aw values as low as
0.6. The alga Dunaliella viridis tolerates sodium chloride concentrations from 1.7
M to a saturated solution.
pH

 pH dramatically affects microbial growth. Each species has a definite pH

growth range and pH growth optimum.

 Acidophiles have their growth optimum between pH 0 and 5.5; neutrophiles,

between pH 5.5 and 8.0; and alkalophiles prefer the pH range of 8.5 to 11.5.
Extreme alkalophiles have growth optima at pH 10 or higher.

 Most bacteria and protozoa are neutrophiles. Most fungi prefer slightly acid
surroundings, about pH 4 to 6; algae also seem to favor slight acidity.

 The alga Cyanidium caldarium and the archaeon Sulfolobus acidocaldarius are
common inhabitants of acidic hot springs; both grow well around pH 1 to 3
and at high temperatures.
 Several mechanisms for the maintenance of a neutral cytoplasmic pH have been
proposed. The plasma membrane may be relatively impermeable to protons.
Neutrophiles appear to exchange potassium for protons using an antiport transport
system.

Ex: Bacillus alkalophilus maintain their internal pH closer to neutrality by exchanging

internal sodium ions for external protons.

 Microorganisms often must adapt to environmental pH changes to survive. In

bacteria, potassium/proton and sodium/proton antiport systems probably correct
small variations in pH.

 If the pH becomes too acidic, other mechanisms come into play. When the pH
drops below about 5.5 to 6.0, Salmonella typhimurium and E. coli synthesize an
array of new proteins as part of what has been called their acidic tolerance
response.

 A proton-translocating ATPase contributes to this protective response, either by

making more ATP or by pumping protons out of the cell. If the external pH
decreases to 4.5 or lower, chaperones such as acid shock proteins and heat shock
proteins are synthesized
Temperature
 Environmental temperature profoundly affects microorganisms, like all other
organisms. Indeed, microorganisms are particularly susceptible because they are
usually unicellular and their temperature varies with that of the external
environment.

 For these reasons, microbial cell temperature directly reflects that of the cell’s
surroundings.

 A most important factor influencing the effect of temperature on growth is the

temperature sensitivity of enzyme catalyzed reactions.

 At low temperatures a temperature rise increases the growth rate because the
velocity of an enzyme-catalyzed reaction, like that of any chemical reaction, will
roughly double for every 10°C rise in temperature.

 Beyond a certain point further increases actually slow growth, and sufficiently
high temperatures are lethal. High temperatures damage microorganisms by
denaturing enzymes, transport carriers, and other proteins. Microbial membranes
are also disrupted by temperature extremes; the lipid bilayer simply melts and
disintegrates.
 Microbial growth has a fairly characteristic temperature dependence with distinct
cardinal temperatures—minimum, optimum, and maximum growth
temperatures.

 One of five classes based on their cardinal temperature ranges for growth.

 Psychrophiles grow well at 0°C and have an optimum growth temperature of 15°C
or lower; the maximum is around 20°C. They are readily isolated from Arctic and
Antarctic habitats.

Ex: The psychrophilic alga Chlamydomonas nivalis with red spores.

 Psychrophiles are widespread among bacterial taxa and found in such genera as
Pseudomonas, Vibrio, Alcaligenes, Bacillus, Arthrobacter, Moritella,
Photobacterium, and Shewanella. The psychrophilic archaeon Methanogenium
has recently been isolated from Ace Lake in Antarctica.

 Psychrophilic microorganisms have adapted to their environment in several ways.

Their enzymes, transport systems, and protein synthetic mechanisms function
well at low temperatures..
The cell membranes of psychrophilic microorganisms have high levels of
unsaturated fatty acids and remain semifluid when cold. Indeed, many
psychrophiles begin to leak cellular constituents at temperatures higher than 20°C
because of cell membrane disruption.

Psychrotrophs or facultative psychrophile

Many species can grow at 0 to 7°C even though they have optima between 20 and
30°C, and maxima at about 35°C.These are called psychrotrophs or facultative
psychrophiles. Psychrotrophic bacteria and fungi are major factors in the spoilage of
refrigerated food.

Mesophiles

Mesophiles are microorganisms with growth optima around 20 to 45°C; they often
have a temperature minimum of 15 to 20°C. Their maximum is about 45°C or lower.
Most microorganisms probably fall within this category. Almost all human
pathogens are mesophiles, as might be expected since their environment is a fairly
constant 37°C.
Thermophiles

 Some microorganisms are thermophiles; they can grow at temperatures of 55°C or

higher. Their growth minimum is usually around 45°C and they often have optima
between 55 and 65°C.

 The vast majority are prokaryotes although a few algae and fungi are thermophilic .
These organisms flourish in many habitats including composts, self-heating hay
stacks, hot water lines, and hot springs.

 Thermophiles differ from mesophiles in having much more heat-stable enzymes

and protein synthesis systems able to function at high temperatures. Their
membrane lipids are also more saturated than those of mesophiles and have
higher melting points; therefore thermophile membranes remain intact at higher
temperatures.

Hyperthermophiles

A few thermophiles can grow at 90°C or above and some have maxima above 100°C.
Procaryotes that have growth optima between 80°C and about 113°C are called
hyperthermophiles.
They usually do not grow well below 55°C. Pyrococcus abyssi and Pyrodictium occultum
are examples of marine hyperthermophiles found in hot areas of the seafloor.

Oxygen Concentration
 Oxygen serves as the terminal electron acceptor for the electron-transport chain in
aerobic respiration. In addition, aerobic eukaryotes employ O2 in the synthesis of
sterols and unsaturated fatty acids.

Micro organisms can be classified into six types based on oxygen need

Aerobe

An organism able to grow in the presence of atmospheric O2

Anaerobe

An organism able to grow in the absence of atmospheric O2

Obligate aerobes
Almost all multicellular organisms are completely dependent on atmospheric O2
for growth.
Facultative anaerobes

Do not require O2 for growth but do grow better in its presence. In the presence
of oxygen they will use aerobic respiration.

Aerotolerant anaerobes

Simply ignore O2 and grow equally well whether it is present or not.

Ex: Enterococcus faecalis

Strict or obligate anaerobes

Do not tolerate O2 at all and die in its presence.

 Bacteroides gingivalis lives in the mouth where it grows in the anaerobic

crevices around the teeth.

 Aerotolerant and strict anaerobes cannot generate energy through respiration

and must employ fermentation or anaerobic respiration pathways for this
purpose.

.
 There are aerobes such as Campylobacter, called microaerophiles, that are
damaged by the normal atmospheric level of O2 (20%) and require O2 levels below
the range of 2 to 10% for growth.

 These different relationships with O2 appear due to several factors, including the
inactivation of proteins and the effect of toxic O2 derivatives. Enzymes can be
inactivated when sensitive groups like sulfhydryls are oxidized. A notable example
is the nitrogen-fixation enzyme nitrogenase which is very oxygen sensitive.

 Oxygen accepts electrons and is readily reduced because its two outer orbital
electrons are unpaired. Flavoproteins several other cell constituents, and radiation
promote oxygen reduction. The result is usually some combination of the
reduction products superoxide radical, hydrogen peroxide, and hydroxyl radical.

 Many microorganisms possess enzymes that afford protection against toxic O2

products. Obligate aerobes and facultative anaerobes usually contain the enzymes
superoxide dismutase (SOD) and catalase, which catalyze the destruction of
superox- ide radical and hydrogen peroxide, respectively. Peroxidase also can be
used to destroy hydrogen peroxide.
 Aerotolerant microorganisms may lack catalase but almost always have
superoxide dismutase. The aerotolerant Lactobacillus plantarum uses
manganous ions instead of superoxide dismutase to destroy the superoxide
radical. All strict anaerobes lack both enzymes or have them in very low
concentrations and therefore cannot tolerate O2.

Pressure

 Most organisms spend their lives on land or on the surface of water, always
subjected to a pressure of 1 atmosphere (atm), and are never affected
significantly by pressure.

 Yet the deep sea (ocean of 1,000 m or more in depth) is 75% of the total ocean
volume. The hydrostatic pressure can reach 600 to 1,100 atm in the deep sea,
while the temperature is about 2 to 3°C.

 Despite these extremes, bacteria survive and adapt. Many are barotolerant:
increased pressure does adversely affect them but not as much as it does
nontolerant bacteria.

 Some bacteria in the gut of deep-sea invertebrates such as amphipods and

holothurians are truly barophilic—they grow more rapidly at high pressures.
 These gut bacteria may play an important role in nutrient recycling in the deep sea.

 One barophile has been recovered from the Mariana trench near the Philippines
(depth about 10,500 m) that is actually unable to grow at pressures below about 400
to 500 atm when incubated at 2°C.

 Thus far, barophiles have been found among several bacterial genera (e.g.,
Photobacterium, Shewanella, Colwellia). Some members of the Archaea are
thermobarophiles (e.g., Pyrococcus spp., Methanococcus jannaschii).

Radiation

 Many forms of electromagnetic radiation are very harmful to microorganisms. This is

particularly true of ionizing radiation, radiation of very short wavelength or high
energy, which can cause atoms to lose electrons or ionize.

 Two major forms of ionizing radiation are (1) X rays, which are artificially produced,
and (2) gamma rays, which are emitted during radioisotope decay.

 Low levels of ionizing radiation will produce mutations and may indirectly result in
death, whereas higher levels are directly lethal. Although microorganisms are more
resistant to ionizing radiation than larger organisms, they will still be destroyed by a
sufficiently large dose.

 Ionizing radiation can be used to sterilize items. Some prokaryotes (e.g., Deinococcus
radiodurans) and bacterial endospores can survive large doses of ionizing radiation.

 A variety of changes in cells are due to ionizing radiation like, it breaks hydrogen
bonds, oxidizes double bonds, destroys ring structures, and polymerizes some
molecules. Oxygen enhances these destructive effects, probably through the
generation of hydroxyl radicals (OH·). Although many types of constituents can be
affected, it is reasonable to suppose that destruction of DNA is the most important
cause of death.

 Ultraviolet (UV) radiation, mentioned earlier, kills all kinds of microorganisms due to
its short wavelength (approximately from 10 to 400 nm) and high energy.

 The most lethal UV radiation has a wavelength of 260 nm, the wavelength most
effectively absorbed by DNA. The primary mechanism of UV damage is the formation
of thymine dimers in DNA . Two adjacent thymines in a DNA strand are covalently
joined to inhibit DNA replication and function.
This damage is repaired in several ways. In photoreactivation, blue light is used by a
photoreactivating enzyme to split thymine dimers. A short sequence containing the
thymine dimer can also be excised and replaced. This process occurs in the absence of
light and is called dark reactivation.

Damage also can be repaired by the recA protein in recombination repair and SOS
repair. When UV exposure is too heavy, the damage is so extensive that repair is
impossible.

Although very little UV radiation below 290 to 300 nm reaches the earth’s surface,
near-UV radiation between 325 and 400 nm can harm microorganisms.

Exposure to near-UV radiation induces tryptophan breakdown to toxic photoproducts.

It appears that these toxic tryptophan photoproducts plus the near-UV radiation itself
produce breaks in DNA strands. The precise mechanism is not known, although it is
different from that seen with 260 nm UV.
Visible light is immensely beneficial because it is the source of energy for
photosynthesis. Yet even visible light, when present in sufficient intensity, can damage
or kill microbial cells. Usually pigments called photosensitizers and O2 are required.

All microorganisms possess pigments like chlorophyll, bacteriochlorophyll,

cytochromes, and flavins, which can absorb light energy, become excited or activated,
and act as photosensitizers. The excited photosensitizer (P) transfers its energy to O2
generating singlet oxygen ( 1 O2).

Singlet oxygen is a very reactive, powerful oxidizing agent that will quickly destroy a
cell. It is probably the major agent employed by phagocytes to destroy engulfed
bacteria.

Many microorganisms that are airborne or live on exposed surfaces use carotenoid
pigments for protection against photooxidation. Carotenoids effectively quench singlet
oxygen—that is, they absorb energy from singlet oxygen and convert it back into the
unexcited ground state. Both photosynthetic and nonphotosynthetic microorganisms
employ pigments in this way