Microbiology Module

Lesson 5. Microbial Growth

The greatest contributor to methods of cultivating bacteria was a German physician,

Robert Koch (1843-1910). Koch was interested in identifying disease-causing bacteria and
experimented with growing bacteria on the cut surfaces of potatoes. He recognized that a single
bacterial cell could multiply in a solid medium in a limited area to form a distinct visible mass of
descendants.

Lesson Outcomes:
1. Describe the stages of growth curve.
2. Describe the general characteristics of bacterial growth.
3. Identify the factors affecting growth and metabolism of microorganisms.

Microorganisms can be found growing even in the harshest climate and

most severe conditions. Each species, however has a limited set of environmental
conditions in which it can grow; even then, it will grow only if specific nutrients
are available. Scientists must be able to grow microorganisms in a culture because
of its medical importance as well as the nutritional and industrial uses. Thus, it is
important to understand the basic principles involved in microbial growth.

Principles of Microbial Growth

Bacteria and archaea generally multiply by the process of binary fission, a process in
which a cell increases its size and then divides. One cell divides into two, those two divides to
become four, those four become eight, and so on. In other words, the increase in cell numbers is
exponential.

Microbial growth is defined as an increased in the number of cells in a population. The

time it takes for a population to double in number is the generation time. This varies greatly from
species to species and is influenced by the conditions in which the cells are grown. For example,
the generation time of a given bacterium may be 30 minutes under ideal conditions, but when the
conditions are not ideal, the organism will grow much more slowly, if at all.

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Fig. 1 Binary fission in bacteria

Source: https://socratic.org/questions/why-is-binary-fission-so-effective-for-bacteria

To calculate how many bacterial cells will be present in a product after a certain amount
of time, two factors must be known initially; the number of cells in the original population and
the number of times the cells will divide during the stated period.

Figure 2 This table shows how many bacteria are in a population that doubles every 20
minutes. The graph is another way to show the same data.
Source: https://bodell.mtchs.org/OnlineBio/BIOCD/text/chapter35/concept35.2.html

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The Growth Curve

Many microorganisms in the laboratory are grown in closed systems, or batch cultures,
because nutrients are not renewed, nor are wastes removed. The growth curve is characterized by
five distinct stages; lag phase, exponential or log phase, stationary phase, death phase, and phase
of prolonged decline.

Fig. 3 Growth Curve.

Source: https://orbitbiotech.com/bacterial-growth-curve-generation-time-lag-phase-log-
phase-exponential-phase-decline-phase/

The lag phase –This is the period of adjustment for the microbes in a new
environment. There is an increase in the size of the microbe but no increase in
number or no cell division during this lag phase. The phase can last from an hour
to several days. The microbes are not dormant but is undergoing extreme
metabolic activity in which enzymes and various molecules are synthesizing.

The log phase- The cells begin to divide and grow or logarithmically increase,
hence the name log phase, or exponential phase. During this period, cellular
activity is occurring as the cell begins to reproduce and generation time is a
constant minimum, thus, the logarithmic plot of growth is a straight line. This
phase is the same in which the cells are most active and production is efficient.
Microorganisms in this phase are particularly sensitive to adverse conditions, such
as radiation and many antimicrobial drugs that can interfere with the growth
process and can harm the microbes.

The stationary phase - Growth rate slows down and microbial deaths balance out
with new cells, stabilizing the population. The metabolic activities of individual
living cells also slow in a period of equilibrium, called the stationary phase. A
number of possible causes to stop the exponential growth may be exhaustion of
nutrients and harmful changes in pH.

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The death phase –This is the period when the total number of viable cells in the
population decreases as cells die off at a constant rate. Deaths eventually exceed
the formation of new cells and the population enters the death phase or the
logarithmic decline phase, where there is the continuous depletion of nutrients
and accumulation of waste materials.

The phase of prolonged decline - A fraction of the cell population survives in the
death phase. These cells have adapted and tolerated the worst conditions, and are
able to multiply for at least a short time, using the nutrients released from the dead
cells. As the conditions continue to deteriorate during the phase of prolonged
decline, most of these survivors then die. However, the few progenies better
equipped for survival can grow, which generates a successive slightly modified
populations, more fit to survive than the previous ones.

What to Do: Answer the following questions

Bacteria multiplication- with Eco the bacterium

This is Eco
Eco is a bacterium
He is going to show you how quickly bacteria can multiply

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1. Plot the information below as a graph. Plot time on the X-axis; the no. of
bacteria on the Y-axis. (5 points).

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2. Indicate on the graph (in no.1) the stages of population cell growth (5
points).

The graph below is the growth curve of your bacteria grown in the media
supplemented with lactose. Use the graph below to answer question no.3. Write
the corresponding letter only.

3.
Your Answers
A. The net increase in population
growth is zero
B.The population growth rate is
constant
C. The cells are not actively dividing
D. The rate of cell growth equals the
rate of cell death
E. The rate of cell death is higher than
the cell growth
F. Essential nutrients are limited
G. Transcription and translation are
actively occurring within the cell
H. Cells are dividing by binary fission
I. DNA replication is occuring

Environmental Factors That Influence Microbial Growth

Major environmental factors that affect microbial growth are temperature,

atmosphere, pH, and water availability.

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Temperature

Each microbial species has a well-defined temperature in which it grows.

Within this range is the optimum growth temperature at which the organism
multiplies most rapidly.
 Psychrophiles – (psychro means “cold”) optimum temperature between -
5°C and 15°C. These organisms grow in the cold Arctic and Antarctic
regions and in lakes fed by glaciers.
 Psychrotrophs – (troph means “nourishment”) optimum temperature
between 15°C and 30°C, but grows well at refrigeration temperature or
lower temperature. They are an important cause of spoilage in refrigerated
foods.
 Mesophiles – (meso means “middle”) optimum temperature between 25°C
and about 45°C. Most medically important bacteria are mesophiles.
 Thermophiles – (thermos means “heat”) optimum temperature between
45°C and 70°C. These organisms commonly live in hot springs and
compost heaps.
 Hyperthermophiles – (hyper means “excessive”) optimum temperature of
70°C or greater. These are usually archaea that are found on the
hydrothermal vent deep in the ocean.

Figure 4. The graph shows growth rate of bacteria as a function of temperature.

Notice that the curves are skewed toward the optimum temperature. The skewing of the growth
curve is thought to reflect the rapid denaturation of proteins as the temperature rises past the
optimum for growth of the microorganism. Source: https://ecampusontario.pressbooks.pub/

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Temperature and Food Preservation

Refrigeration temperatures approximately 4°C slows spoilage because they

limit the multiplication of fast-growing mesophiles. Psychrophiles and
psychrotrophs can still grow at these temperatures, and refrigerated foods will still
spoil, but more slowly.
Foods and other perishable products that withstand below-freezing
temperatures can be frozen for long term storage, though freezing is not an
effective means of destroying microbes. Freezing is routinely used to preserve
stock cultures.

Temperature and Disease

The temperatures of different parts of the human body vary significantly.

For example, the brain, heart, and gastrointestinal tract are near 37°C, but the
temperature of the extremities is lower. For this reason, some microbes can cause
disease more readily in certain parts of the body.

Oxygen Requirements

Oxygen requirement/tolerance reflects the organism’s energy-harvesting

mechanisms and its ability to inactivate reactive oxygen species. Aerobic, means
microorganisms require oxygen. Anaerobic, means microorganisms thrive in
environments that have little or no oxygen present.
 Obligate aerobes – have an absolute requirement for
oxygen. Ex: Micrococcus luteus, which is common
in the environment.
 Facultative anaerobes – grow better if oxygen is present, but can also
grow without it. The term “facultative” means that the organism is flexible,
in its requirements for O2. Ex: E. coli, grows faster when O2 is present and
most commonly found in the large intestine.
 Obligate anaerobes – cannot grow in the presence of O 2, they are often
killed by even brief exposure to air. Ex: Clostridium botulinum, that causes
botulism.
 Microaerophiles – require small amount of O2 (2% to 10%). Ex: Helicobacter
pylori, which causes gastric and duodenal ulcers.
 Aerotolerant anaerobes – indifferent to O2 ,also called obligate
fermenters because fermentation is their only metabolic option. Ex:
Streptococcus pyogenes, which causes strep throat.

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pH (Acidity)

Each microbial species can survive within a range of pH values, which is

pH optimum. Despite the pH of the external environment, cells maintain a
constant internal pH, typically near neutral. Many prokaryotes that grow in acidic
environments quickly pump out protons that enter the cell.
 Neutrophiles - Live and multiply within the range of pH 5 (acidic) to pH 8
(basic), and have a pH optimum near neutral (pH 7). Food preservation
methods such as pickling inhibit bacterial growth by increasing the acidity
of the food.
 Acidophiles - grow optimally at a pH below 5.5. Ex: Picrophilus oshimae
(Archaea), has an optimum pH of less than 1.
 Alkaliphiles – grow optimally at a pH above 8.5. They often live in
alkaline lakes and soils.

Growth of any microorganism depends not only on a suitable physical environment, but
also on the availability of nutrients, which the cell used for biosynthesis. There are two
categories of essential nutrients: macro-nutrients (which are needed in large amounts) and micro-
nutrients (which are needed in trace or small amounts). Macro-nutrients usually help maintain
the cell structure and metabolism. Micro-nutrients help enzyme function and maintain protein
structure.

Essential Nutrients

The sources of common essential nutrients are carbon, hydrogen, nitrogen, oxygen, phosphorus,
and sulfur. Organisms usually absorb carbon when it is in its organic form.

 Carbon in its organic form is usually a product of living things.

 Another essential nutrient, nitrogen, is part of the structure of

protein, DNA, RNA, and ATP. Nitrogen is important for heterotroph survival, but it must

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first be degraded into basic building blocks, such as amino acids, in order to be used.
Some microorganisms use nitrogen gas (N2) as a nitrogen source, converting it to
ammonia and then incorporating that into cellular material.

 Oxygen is an important component of both organic and inorganic

compounds. It is essential to the metabolism of many organisms.

 Hydrogen has many important jobs including maintaining the pH of

solutions and providing free energy in reactions of respiration.

 Phosphate is an important player in making nucleic acids and cellular

energy transfers. Without sufficient phosphate, an organism will cease to grow.

 Lastly, sulfur is found in rocks and sediments and is found widely in mineral
form. Many microbes use Inorganic sulfur sources such as sulfate, but others require
organic sources such as sulfur-containing amino acids.

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What to Do: Answer the following questions

Who am I?

1. A left -over food from the birthday celebration of Rhieselle was forgotten in the
refrigerator and shows contamination. The contaminants are probably which of the
following?
a. Thermophiles
b. Acidophiles
c. Psychotrophs
d. Mesophiles

2. Bacteria isolated from a hot tub at 39’C are probably which of the following?
a. Thermophiles
b. Mesophiles
c. Psychotrophs
d. Hyperthermophiles

3. In which environment are you most likely to encounter a hyperthemophile?

a. Hot tub
b. Human body
c. Hydrothermal vent at the bottom of the ocean
d. Warm ocean water in Dubai

4. Which of the following environments would harbor psychrophiles?

a. Contaminated glass left in a 35’C incubator
b. Mountain river with a water temperature of 12’C.
c. Yoghurt cultured at room temperature
d. Salt pond in a desert with a daytime temperature of 34’C.

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5. Thermus aquaticus and Geobacillus can grow well in temperature range of 80’C to a
maximum of 110’C. What type of microbes are they?
a. Mesophiles
b. Hyperthermophiles
c. Psychrophiles
d. Acidophiles
6. Pathogenic microbes are:
a. Mesophiles
b. Psychrophiles
c. Psychrotrophs
d. Thermophiles

References:
Anderson, D.G., S.N. Salm, D.P. Allen and E.W. Nester. (2016). Nester’s Microbiology. 8 th ed.
Mc-Graw Hill Education, USA
https://www.britannica.com/science/bacteria/Growth-of-bacterial-populations
https://bodell.mtchs.org/OnlineBio/BIOCD/text/chapter35/concept35.2.html
https://courses.lumenlearning.com/boundless-microbiology/chapter/microbial-nutrition/
https://courses.lumenlearning.com/microbiology/chapter/temperature-and-microbial-growth/
https://ecampusontario.pressbooks.pub/microbio/chapter/temperature-and-microbial-growth/

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