Talaro−Talaro: Foundations 7.
Elements of Microbial Text
in Microbiology, Fourth Nutrition, Ecology, and
Edition Growth
400ⴛ Oil droplet
Pseudopod
Vacuole
Cell
Phagocytosis
FIGURE 7.8
Endocytosis (phagocytosis and pinocytosis). Solid particles are phagocytosed by large cell extensions called pseudopods, and they are
pinocytosed into vesicles by very fine cell protrusions called microvilli. Oil droplets fuse with the membrane and are released directly into the cell.
ously forming a vacuole and engulfing it (figure 7.8). Amebas and
certain white blood cells ingest whole cells or large solid matter by
phagocytosis.* Liquids, such as oils or large molecules in solution,
enter the cell through pinocytosis.*
*phagocytosis (fag-oh-cy-toh-sis) Gr. phagein, to eat.
*pinocytosis (pin-oh-cy-toh-sis) Gr. pino, to drink.
CHAPTER CHECKPOINTS
Nutrition is a process by which all living organisms obtain substances
from their environment to convert to metabolic uses.
Although the chemical form of nutrients varies widely, all organisms
require six bioelements—carbon, hydrogen, oxygen, nitrogen,
phosphorus, and sulfur—to survive, grow, and reproduce.
Nutrients are categorized by the amount required (macronutrients or
micronutrients), by chemical structure (organic or inorganic), and by
their importance to the organism’s survival (essential or nonessential).
Microorganisms are classified both by the chemical form of their nutrients
and the energy sources they utilize.
Nutrient requirements of microorganisms determine their respective
niches in the food webs of major ecosystems.
Nutrients are transported into microorganisms by two kinds of processes:
active transport that expends energy and passive transport that occurs
independently of energy input.
The molecular size and concentration of a nutrient determine which method
of transport is used.
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Environmental Factors That Influence Microbes 199
1,000ⴛ
Microvillus
Enclosure
of liquid
between
microvilli
Pinocytosis
Liquid
in vesicle
Environmental Factors That
Influence Microbes
Microbes are exposed to a wide variety of environmental factors in
addition to nutrients. Microbial ecology focuses on ways that mi-
croorganisms deal with or adapt to such factors as heat, cold, gases,
acid, radiation, osmotic and hydrostatic pressures, and even other
microbes. Adaptation is a complex adjustment in biochemistry or
genetics that enables long-term survival and growth. For most mi-
crobes, environmental factors fundamentally affect the function of
metabolic enzymes. Thus, survival in a changing environment is
largely a matter of whether the enzyme systems of microorganisms
can adapt to alterations in their habitat. Incidentally, one must be
careful to differentiate between growth in a given condition and
tolerance, which implies survival without growth.
TEMPERATURE ADAPTATIONS
Microbial cells are unable to control their temperature and therefore
assume the ambient temperature of their natural habitats. Their sur-
vival is dependent on adapting to whatever temperature variations
are encountered in that habitat. The range of temperatures for mi-
crobial growth can be expressed as three cardinal temperatures. The
minimum temperature is the lowest temperature that permits a mi-
crobe’s continued growth and metabolism; below this temperature,
its activities are inhibited. The maximum temperature is the high-
est temperature at which growth and metabolism can proceed. If the
temperature rises slightly above maximum, growth will stop, but if
 Talaro−Talaro: Foundations 7. Elements of Microbial Text © The McGraw−Hill
in Microbiology, Fourth Nutrition, Ecology, and Companies, 2002
Edition Growth

200 CHAPTER 7 Elements of Microbial Nutrition, Ecology, and Growth

it continues to rise beyond that point, the enzymes and nucleic acids Psychrophile
will eventually become permanently inactivated and the cell will Mesophile
Optimum
die. This is why heat works so well as an agent in microbial control. Thermophile
The optimum temperature covers a small range, intermediate be-

Rate of Growth
tween the minimum and maximum, which promotes the fastest rate
of growth and metabolism (rarely is the optimum a single point).
Depending on their natural habitats, some microbes have a
narrow cardinal range, others a broad one. Some strict parasites will
not grow if the temperature varies more than a few degrees below or
Minimum Maximum
above the host’s body temperature. For instance, the typhus rick-
ettsia multiplies only in the range of 32°–38°C, and rhinoviruses
(one cause of the common cold) multiply successfully only in tis-
sues that are slightly below normal body temperature (33°–35°C). -15 -10 -5 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90
Other parasites are not so limited. Strains of Staphylococcus aureus Temperature °C
grow within the range of 6°–46°C, and the intestinal bacterium
FIGURE 7.9
Enterococcus faecalis grows within the range of 0°–44°C.
Another way to express temperature adaptation is to describe Ecological groups by temperature of adaptation. Psychrophiles
can grow at or near 0°C and have an optimum below 15°C. As a
whether an organism grows optimally in a cold, moderate, or hot group, mesophiles can grow between 10°C and 50°C, but their optima
temperature range. The terms used for these ecological groups are usually fall between 20°C and 40°C. Generally speaking, thermophiles
psychrophile, mesophile, and thermophile (figure 7.9), respectively. require temperatures above 45°C and grow optimally between this
A psychrophile (sy-kroh-fyl) is a microorganism that has an temperature and 80°C. The cardinal temperatures are labeled for
optimum temperature below 15°C and is capable of growth at 0°C. mesophiles. Note that the extremes of the ranges can overlap to an
extent.
It is obligate with respect to cold and generally cannot grow above
20°C. Laboratory work with true psychrophiles can be a real chal-
lenge. Inoculations have to be done in a cold room because room
temperature can be lethal to the organisms. Unlike most laboratory
cultures, storage in the refrigerator incubates, rather than inhibits,
them. As one might predict, the habitats of psychrophilic bacteria,
fungi, and algae are snowfields (figure 7.10), polar ice, and the deep
ocean. Rarely, if ever, are they pathogenic. True psychrophiles must
be distinguished from psychrotrophs or facultative psychrophiles
that grow slowly in cold but have an optimum temperature above
20°C. Bacteria such as Staphylococcus aureus and Listeria mono-
cytogenes are a concern because they can grow in refrigerated food
and cause food-borne illness.
The majority of medically significant microorganisms are
mesophiles (mez-oh-fylz), organisms that grow at intermediate (a)
temperatures. Although an individual species can grow at the ex-
tremes of 10°C or 50°C, the optimum growth temperatures (op-
tima) of most mesophiles fall into the range of 20°–40°C. Organ-
isms in this group inhabit animals and plants as well as soil and
water in temperate, subtropical, and tropical regions. Most human
pathogens have optima somewhere between 30°C and 40°C (hu-
man body temperature is 37°C). Thermoduric microbes, which can
survive short exposure to high temperatures but are normally
mesophiles, are common contaminants of heated or pasteurized
foods (see chapter 11). Examples include heat-resistant cysts such
as Giardia or sporeformers such as Bacillus and Clostridium.
A thermophile (thur-moh-fyl) is a microbe that grows opti-
mally at temperatures greater than 45°C. Such heat-loving mi-
crobes live in soil and water associated with volcanic activity and
in habitats directly exposed to the sun. Thermophiles vary in heat (b)
requirements, with a general range of growth of 45°–80°C. Most
FIGURE 7.10
eucaryotic forms cannot survive above 60°C, but a few ther-
Red snow. (a) An early summer snowbank provides a perfect habitat
mophilic bacteria called hyperthermophiles, grow between 80°C for psychrophilic photosynthetic organisms like Chlamydomonas nivalis.
and 110°C (currently thought to be the temperature limit endured (b) Microscopic view of this snow alga (actually classified as a “green”
by enzymes and cell structures). Strict thermophiles are so heat- alga).
 Talaro−Talaro: Foundations 7. Elements of Microbial Text
in Microbiology, Fourth Nutrition, Ecology, and
Edition Growth
SPOTLIGHT ON MICROBIOLOGY
Cashing in on “Hot” Microbes
The smoldering thermal springs in Yellowstone National Park are more
than just one of the geologic wonders of the world. They are also a hotbed
of some of the most unusual microorganisms in the world. The ther-
mophiles thriving at temperatures near the boiling point are the focus of
serious interest from the scientific community. For many years, biologists
have been intrigued that any living thing could function at such high tem-
peratures. Such questions as these come to mind: Why don’t they melt
and disintegrate, why don’t their proteins coagulate, and how can their
DNA possibly remain intact?
One of the earliest thermophiles to be isolated was Thermus aquati-
cus. It was discovered by Thomas Brock in Yellowstone’s Mushroom Pool
in 1965 and was registered with the American Type Culture Collection. In-
terested researchers studied this species and discovered that it has ex-
tremely heat-stable proteins and nucleic acids, and its cell membrane does
not break down readily at high temperatures. Later, an extremely heat-
stable DNA-replicating enzyme was isolated from the species.
What followed is a riveting example of how pure research for the
sake of understanding and discovery also offered up a key ingredient in a
multimillion-dollar process. Developers of the polymerase chain reaction
(PCR), a versatile tool for making multiple copies of DNA fragments,
found that the technique would work only if they performed the test at
temperatures around 65°–72°C. The mesophilic enzymes they tested
were destroyed at such high temperatures, but the Thermus DNA-
copying enzyme (Taq polymerase) functioned splendidly. The PCR tech-
nique became the basis for a variety of test procedures in forensics and
gene detection and analysis.
Spurred by this remarkable success story, biotechnology compa-
nies have descended on Yellowstone, which contains over 10,000 hot
springs, geysers, and hot habitats. These industries are looking to unusual
bacteria and archaea as a source of “extremozymes,” enzymes that oper-
tolerant that researchers may use an autoclave to isolate them in
culture. Currently, there is intense interest in thermal microorgan-
isms by biotechnology companies (Spotlight on Microbiology 7.4).
GAS REQUIREMENTS
The atmospheric gases that most influence microbial growth are
O2 and CO2. Of these, oxygen gas has the greatest impact on mi-
crobial adaptation. Not only is it an important respiratory gas, but
it is also a powerful oxidizing agent that exists in many toxic
forms. In general, microbes fall into one of three categories: those
that use oxygen and can detoxify it; those that can neither use oxy-
gen nor detoxify it; and those that do not use oxygen but can
detoxify it.
How Microbes Process Oxygen
As oxygen enters into cellular reactions, it is transformed into sev-
eral toxic products. Singlet oxygen (1O2) is an extremely reactive
molecule produced by both living and nonliving processes. No-
© The McGraw−Hill
Companies, 2002
Environmental Factors That Influence Microbes 201
7.4
Biotechnology researchers harvesting samples in Yellowstone
National Park.
ate under high temperatures and acidity. So far, about a dozen other
organisms with useful enzymes have been discovered. They may have
applications in developing high-temperature fermentations, cleaning up
toxic wastes, and organic syntheses.
This quest has also brought attention to questions such as: Who
owns these microbes, and can their enzymes be patented? As of the year
2000, the Park Service has gotten a legal ruling that allows them to share
in the profits from companies and to add that money to their operating
budget. The U.S. Supreme Court has also ruled that a microbe isolated
from natural habitats cannot be patented. Only the technology that uses
the microbe can be patented.
tably, it is one of the substances produced by phagocytes to kill
invading bacteria (see chapter 14). The buildup of singlet oxygen
and the oxidation of membrane lipids and other molecules can
damage and destroy a cell. The highly reactive superoxide ion
(O2), peroxides (H2O2), and hydroxyls (OH) are other destruc-
tive metabolic by-products of oxygen. To protect themselves
against damage, most cells have developed enzymes that go about
the business of scavenging and neutralizing these chemicals. The
complete conversion of superoxide ion into harmless oxygen
requires a two-step process and at least two enzymes:
Superoxide
Step 1. O2
O2
2H
dismutase
H2O2 (hydrogen peroxide)
O2
Step 2. H2O2
H2O2 Catalase
2H2O
O2
In this series of reactions (essential for aerobic organisms),
the superoxide ion is first converted to hydrogen peroxide and
normal oxygen by the action of an enzyme called superoxide
dismutase. Because hydrogen peroxide is also toxic to cells (it is a
 Talaro−Talaro: Foundations 7. Elements of Microbial Text © The McGraw−Hill
in Microbiology, Fourth Nutrition, Ecology, and Companies, 2002
Edition Growth

202 CHAPTER 7 Elements of Microbial Nutrition, Ecology, and Growth

disinfectant and antiseptic), it will be degraded by the enzyme

catalase into water and oxygen. If a microbe is not capable of
dealing with toxic oxygen by these or similar mechanisms, it is
forced to live in habitats free of oxygen.
With respect to oxygen requirements, several general cate-
gories are recognized. An aerobe* (aerobic organism) can use
gaseous oxygen in its metabolism and possesses the enzymes
needed to process toxic oxygen products. An organism that cannot
grow without oxygen is an obligate aerobe. Most fungi and proto-
zoa, as well as many bacteria (genera Micrococcus and Bacillus),
have strict requirements for oxygen in their metabolism.
A facultative anaerobe is an aerobe that does not require
oxygen for its metabolism and is capable of growth in the absence
of oxygen. This type of organism metabolizes by aerobic respira-
tion when oxygen is present, but in its absence, it adopts an anaer-
obic mode of metabolism such as fermentation. Facultative anaer- (a)
obes usually possess catalase and superoxide dismutase. A large Lockscrew
number of bacterial pathogens fall into this group (for example,
Outer lid Inner lid Catalyst chamber contains
gram-negative enteric* bacteria and staphylococci). A microaero- palladium pellets.
phile (myk-roh-air-oh-fyl) does not grow at normal atmospheric
tensions of oxygen but requires a small amount of it in metabolism.
2H2 + O2 2H2O
Most organisms in this category live in a habitat (soil, water, or the Rubber gasket
CO2 H2
human body) that provides small amounts of oxygen but is not di- provides air-tight seal.
rectly exposed to the atmosphere.
An anaerobe (anaerobic microorganism) lacks the metabolic Gas Pack

enzyme systems for using oxygen in respiration. Because strict, or Gas generator envelope
obligate, anaerobes also lack the enzymes for processing toxic oxy- (10 ml of water is added to
chemicals in envelope to
gen, they cannot tolerate any free oxygen in the immediate environ- generate H2 and CO2.
ment and will die if exposed to it. Strict anaerobes live in highly re- BBL Carbon dioxide promotes
duced habitats, such as deep muds, lakes, oceans, and soil. Even more rapid growth of organisms.)
though human cells use oxygen and oxygen is found in the blood and Anaerobic indicator strip Reaction
(Methylene blue becomes colorless (Oxygen is removed from chamber
tissues, some body sites present anaerobic pockets or microhabitats by combining with hydrogen on
in absence of O2.)
where colonization or infection can occur. One region that is an im- surface of palladium pellets.)
(b)
portant site for anaerobic infections is the oral cavity. Dental caries
are partly due to the complex actions of aerobic and anaerobic bacte- FIGURE 7.11
ria, and most gingival infections consist of similar mixtures of oral Culturing techniques for anaerobes. (a) A special anaerobic
bacteria that have invaded damaged gum tissues. Another common environmental chamber makes it possible to handle strict anaerobes
site for anaerobic infections is the large intestine, a relatively oxygen- without exposing them to air. It also has provisions for incubation and
inspection in a completely O2-free system. (b) The anaerobic jar, or CO2
free habitat that harbors a rich assortment of strictly anaerobic bacte-
incubator system. To create an anaerobic environment, a packet is
ria. Anaerobic infections can accompany abdominal surgery and activated to produce hydrogen gas, and the chamber is sealed tightly.
traumatic injuries (gas gangrene and tetanus). Growing anaerobic The gas reacts with available oxygen to produce water. Carbon dioxide
bacteria usually requires special media, methods of incubation, and can also be added to the system for growth of capnophiles.
handling chambers that exclude oxygen (figure 7.11a).
Aerotolerant anaerobes do not utilize oxygen but can sur-
vive and grow to a limited extent in its presence. These anaerobes (those that contain an oxygen-absorbing chemical). One such tech-
are not harmed by oxygen, mainly because they possess alternate nique demonstrates oxygen requirements by the location of growth
mechanisms for breaking down peroxides and superoxide. Certain in a tube of fluid thioglycollate (figure 7.12).
lactobacilli and streptococci use manganese ions or peroxidases to Although all microbes require some carbon dioxide in their
perform this task. metabolism, capnophiles* grow best at a higher CO2 tension than is
Determining the oxygen requirements of a microbe from a normally present in the atmosphere. This becomes important in the
biochemical standpoint can be a very time-consuming process. Of- initial isolation of some pathogens from clinical specimens, notably
ten it is illuminating to perform culture tests with reducing media Neisseria (gonorrhea, meningitis), Brucella (undulant fever), and
Streptococcus pneumoniae. Incubation is carried out in a CO2 incu-
*aerobe (air-ohb) Although the prefix means air, it is used in the sense of oxygen. bator that provides 3% to 10% CO2 (see figure 7.11b).
*enteric (en-terr-ik) Gr. enteron, intestine. A family of bacteria that live in the large
intestines of animals. *capnophile (kap´-noh-fyl) Gr. kapnos, smoke.
 Talaro−Talaro: Foundations 7. Elements of Microbial Text
in Microbiology, Fourth Nutrition, Ecology, and
Edition Growth
Demonstration of Oxygen Requirements
High
O2
tension
Low
Aerobic Microaerophilic Facultative Aerotolerant Anaerobic
(top growth) (growth just anaerobic anaerobic (bottom
below surface) (growth (some growth growth)
throughout) in O2)
FIGURE 7.12
Use of thioglycollate broth to demonstrate oxygen requirements.
Thioglycollate is a chemical that absorbs O2 gas from the air. Oxygen at
the top of the tube is dissolved in the medium and absorbed; its
presence is indicated by the red dye resazurin. When a series of tubes
is inoculated with bacteria that differ in O2 requirements, the relative
position of growth provides some indication of their adaptations to
oxygen use.
EFFECTS OF pH
Microbial growth and survival are also influenced by the pH of the
habitat. The pH was defined in chapter 2 as the degree of acidity or
alkalinity (basicity) of a solution. It is expressed by the pH scale, a
series of numbers ranging from 0 to 14. The pH of pure water (7.0)
is neutral, neither acidic nor basic. As the pH value decreases to-
ward 0, the acidity increases, and as the pH increases toward 14, the
alkalinity increases. The majority of organisms live or grow in
habitats between pH 6 and 8 because strong acids and bases can be
highly damaging to enzymes and other cellular substances.
A few microorganisms live at pH extremes. Obligate acid-
ophiles include Euglena mutabilis, an alga that grows in acid pools
between 0 and 1.0 pH, and Thermoplasma, an archaea that lacks a
cell wall, lives in hot coal piles at a pH of 1 to 2, and will lyse if ex-
posed to pH 7. Because many molds and yeasts tolerate moderate
acid, they are the most common spoilage agents of pickled foods.
Alkalinophiles live in hot pools and soils that contain high levels of
basic minerals (up to pH 10.0). Bacteria that decompose urine cre-
ate alkaline conditions, since ammonium (NH4
, an alkaline ion)
can be produced when urea (a component of urine) is digested. Me-
tabolism of urea is one way that the ulcer bacterium Helicobacter
pylori can neutralize the acidity of the stomach.
OSMOTIC PRESSURE
Although most microbes exist under hypotonic or isotonic condi-
tions, a few, called halophiles (hay-loh-fylz), live in habitats
with a high solute concentration. Obligate halophiles such as
Halobacterium and Halococcus inhabit salt lakes, ponds, and
other hypersaline habitats. They grow optimally in solutions of
© The McGraw−Hill
Companies, 2002
Environmental Factors That Influence Microbes 203
25% NaCl but require at least 9% NaCl (combined with other
salts) for growth. These archaea have significant modifications in
their cell walls and membranes and will lyse in hypotonic habi-
tats. Facultative halophiles are remarkably resistant to salt, even
though they do not normally reside in high-salt environments.
For example, Staphylococcus aureus can grow on NaCl media
ranging from 0.1% up to 20%. Although it is common to use high
concentrations of salt and sugar to preserve food ( jellies, syrups,
and brines), many bacteria and fungi actually thrive under these
conditions and are common spoilage agents. The term to describe
microbes that withstand and grow at high osmotic pressures is
osmophile.
MISCELLANEOUS ENVIRONMENTAL FACTORS
Various forms of electromagnetic radiation (ultraviolet, infrared,
visible light) stream constantly onto the earth from the sun. Some
microbes (phototrophs) can use visible light rays as an energy
source, but non-photosynthetic microbes tend to be damaged by the
toxic oxygen products produced by contact with light. Some mi-
crobial species 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 (X rays and cosmic rays). In chap-
ter 11, we will see just how these types of energy are applied in mi-
crobial control.
Descent into the ocean depths subjects organisms to increas-
ing hydrostatic pressure. Deep-sea microbes called barophiles ex-
ist under pressures that range from a few times to over 1,000 times
the pressure of the atmosphere. These bacteria are so strictly
adapted to high pressures that they will rupture when exposed to
normal atmospheric pressure.
Because of the high water content of cytoplasm, all cells re-
quire water from their environment to sustain growth and metabo-
lism. Water is the solvent for cell chemicals, and it is needed for en-
zyme function and digestion of macromolecules. A certain amount
of water on the external surface of the cell is required for the diffu-
sion of nutrients and wastes. Even in apparently dry habitats, such
as sand or dry soil, the particles retain a thin layer of water usable
by microorganisms. Dormant, dehydrated cell stages (for example,
spores and cysts) tolerate extreme drying because of the inactivity
of their enzymes.
ECOLOGICAL ASSOCIATIONS AMONG
MICROORGANISMS
Up to now, we have considered the importance of nonliving envi-
ronmental influences on the growth of microorganisms. Another
profound influence comes from other organisms that share (or
sometimes are) their habitats. In all but the rarest instances,
microbes live in shared habitats, which give rise to complex and
fascinating associations. Some associations are between similar or
dissimilar types of microbes; others involve multicellular organ-
isms such as animals or plants. Interactions can have beneficial,
harmful, or no particular effects on the organisms involved; they
can be obligatory or nonobligatory to the members; and they often