5/20/2016

MICROBIAL METABOLISM AND

MICROBIAL GROWTH RATE

BIO149 Prof. UREAH THEA A. SEVILLA

Metabolism
 Metabolism is the intricately regulated system of
energy-producing and energy-utilizing chemical
reactions in an organism.

Two Processes:
- ANABOLISM

- CATABOLISM

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Metabolism

Anabolism Catabolism
- biosynthesis - degradation
- diverging process - converging process
- reductive process - oxidative process
- endergonic process - exergonic process

Degradative and Biosynthetic Pathways

large energy-rich
molecules

ADP
+ Pi BIOSYNTHETIC
DEGRADATIVE PATHWAYS
PATHWAYS (ANABOLIC)
(CATABOLIC)
ATP
simple organic
energy-poor compounds
products
ENERGY INPUT

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Three Principles in Microbial Metabolism

1. How the organism obtains carbon for synthesizing

cell mass:
 Autotrophic – carbon obtained from CO2
 Heterotrophic – carbon obtained from carbon
compounds
 Mixotrophic – carbon obtained from CO2 and carbon
compounds

Three Principles in Microbial Metabolism

2. How the organism obtains reducing equivalents

used either in energy conservation or biosynthetic
reactions:
 Lithotrophic – reducing equivalents obtained from
inorganic compounds
 Organotrophic – reducing equivalents obtained from
organic compounds

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Three Principles in Microbial Metabolism

3. How the organism obtains energy for living and

growing
 Chemotrophic – energy is obtained from external
chemical compounds
 Phototrophic – energy is obtained from light

Microbial Growth
 Refers to an increase in cell number, not in cell size.
 Bacteria grow and divide by binary fission, a
rapid and relatively simple process.

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Microbial Growth
4 Phases:
• Lag Phase

• Log Phase

• Stationary Phase

• Death

Microbial Growth
LAG PHASE
 Period of adjustment to
new conditions.
 Little or no cell division
occurs, population size
 doesn’t increase.
 Phase of intense metabolic LAG

activity, in which PHASE

 individual organisms grow

in size.
 May last from one hour to
several days.

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Microbial Growth
LOG PHASE
 Cells begin to divide and
generation time reaches
a constant minimum.
 Period of most rapid
growth. LOGARITHMIC
PHASE

 Cells are at highest

metabolic activity.
 Cells are most susceptible
to adverse environmental
factors at this stage.
 • Radiation
 • Antibiotics Number of cells produced > Number of cells dying

Microbial Growth
STATIONARY PHASE
STATIONARY PHASE
 Population size begins to
stabilize.
 Overall cell number does not
increase.
 Cell division begins to slow
down.
 Factors that slow down
microbial growth:
• Accumulation of toxic waste
materials
• Acidic pH of media
• Limited nutrients Number of cells produced = Number of cells dying
• Insufficient oxygen supply

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Generation Time
 The rate of exponential growth of a bacterial culture is
expressed as Generation Time, also the Doubling time
of the bacterial population.

Generation time (G) is defined as the time (t) per

generation (n = number of generations)
G= t / n

It is the time interval required for the cells (or population)

to divide.

Generation Times of Common Bacteria

Generation times for some common bacteria under optimal conditions of
growth.
Generation Time
Bacterium Medium
(minutes)
Escherichia coli Glucose-salts 17
Bacillus megaterium Sucrose-salts 25
Streptococcus lactis Milk 26
Streptococcus lactis Lactose broth 48
Staphylococcus aureus Heart infusion broth 27-30
Lactobacillus acidophilus Milk 66-87
Mannitol-salts-yeast
Rhizobium japonicum 344-461
extract
Mycobacterium
Synthetic 792-932
tuberculosis
Treponema pallidum Rabbit testes 1980

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Expression of Bacterial Growth

This equation is an expression of growth by binary
fission: b = B x 2n

where
b = number of bacteria at the end of time interval
B = number of bacteria at the start of the time interval
n = number of generations (number of times the cell
population doubles during the time interval)

Sample Problem
What is the generation time of a bacterial
population that increases from 10,000 cells to
10,000,000 cells in four hours of growth?

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Microbial Growth
DEATH OR DECLINE PHASE:
 Population size begins to
decrease. DEATH
 Cell number decreases at
a logarithmic rate.
 Cells lose their ability to
divide.
 A few cells may remain
alive for a long period of
time.
Number of cells produced < Number of cells dying

Environmental Factors Affecting

Microbial Growth

 Temperature
 pH
 Water and Osmotic Pressure
 Oxygen Concentration

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Growth Rate of Bacteria Groups at Different

Temperature

Microbes are loosely classified into several

groups based on their preferred temperature
ranges.

Growth Rate of Bacteria Groups at Different

Temperature

Psychrophiles: “Cold-loving”. Can grow at 0C. Two

groups:
True Psychrophiles: Sensitive to temperatures
over 20C. Optimum growth at 15C or below.
Found in very cold environments (North pole,
ocean depths).
 Seldom cause disease or food spoilage.
Psychrotrophs: Optimum growth at 20 to 30C.
 Responsible for most low temperature food
spoilage.

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Growth Rate of Bacteria Groups at Different

Temperature

Psychrophiles: “Cold-loving”. Can grow at 0C. Two

groups:
True Psychrophiles: Sensitive to temperatures
over 20C. Optimum growth at 15C or below.
Found in very cold environments (North pole,
ocean depths).
 Seldom cause disease or food spoilage.
Psychrotrophs: Optimum growth at 20 to 30C.
 Responsible for most low temperature food
spoilage.

Growth Rate of Bacteria Groups at Different

Temperature

Mesophiles: “Middle loving”. Most

bacteria.
 Include most pathogens and common
spoilage organisms.
 Best growth between 25 to 40C.

 Optimum temperature commonly

37C.
 Many have adapted to live in the
bodies of animals.

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Growth Rate of Bacteria Groups at Different

Temperature

Thermophiles: “Heat loving”.

 Optimum growth between 50 to

60

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Bacterial Groups at Different pH

Organisms can be classified as:
A. Acidophiles: “Acid loving”.
 Grow at very low pH (0.1 to 5.5)
 Lactobacillus produces lactic acid, tolerates mild acidity.
B. Neutrophiles:
 Grow at pH 5.5 to 8.5.
 Includes most human pathogens.
C. Alkaliphiles: “Alkali loving”.
 Grow at alkaline or high pH (7 to 12 or higher)
 Vibrio cholerae and Alkaligenes faecalis optimal pH 9.
 Soil bacterium Agrobacterium grows at pH 12.

Water and Osmotic Pressure influence

on Microbial Growth
 Solutes and water activity (a quantitative measurement
of the availability of water) is inversely related to
osmotic pressure and may have a profound effect on
cell growth
 Osmotolerant organisms can grow in solutions of both high
and low water activity
 Halophiles require environments of low water activity (high
osmotic pressure) in order to grow
 Microorganisms growing in a habitat with low water activity
(aw) usually maintain a high internal solute concentration in
order to retain water

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Influence of Oxygen Requirement on

Microbial Growth
1. Obligate aerobes are completely dependent on
atmospheric O2 for growth
2. Facultative anaerobes do not require O2 for growth,
but do grow better in its presence
3. Aerotolerant anaerobes ignore O2 and grow equally
well whether it is present or not
4. Obligate (strict) anaerobes do not tolerate O2 and die
in its presence
5. Microaerophiles are damaged by the normal
atmospheric level of O2 (20%) but require lower levels
(2 to 10%) for growth

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Chemical Requirement for Growth

 Microorganisms are cultured in water to which
appropriate dissolved nutrients are added.
 These nutrients fall into three categories:
 Energy sources
 Cell structural components (Elemental Requirements)
 Miscellaneous Growth factors not all kinds of
organisms require the same nutrients nor can any
one organism use all kinds.

Methods for Measurement of Cell Numbers

Measuring techniques involve:

 Direct counts, visually or instrumentally

 Indirect viable cell counts

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Methods for Measurement of Cell Numbers

 Direct microscopic counts are possible using special

slides known as counting chambers. Dead cells cannot
be distinguished from living ones. Only dense
suspensions can be counted (>107 cells per ml), but
samples can be concentrated by centrifugation or
filtration to increase sensitivity.
 Electronic counting chambers count numbers and
measure size distribution of cells. For cells the size of
bacteria the suspending medium must be very clean.
Such electronic devices are more often used to count
eukaryotic cells such as blood cells.

Methods for Measurement of Cell Numbers

 Indirect viable cell counts, also called plate counts,

involve plating out (spreading) a sample of a culture
on a nutrient agar surface. The sample or cell
suspension can be diluted in a nontoxic diluent (e.g.
water or saline) before plating. If plated on a
suitable medium, each viable unit grows and forms a
colony. Each colony that can be counted is called a
Colony Forming Unit (cfu) and the number of cfu's is
related to the viable number of bacteria in the
sample.

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Measurement of Bacterial Growth

A standard plate count

reflects the number of viable
microbes and assumes that
each bacterium grows into a
single colony. Because it is
impossible to say that each
colony actually arose from
an indivual cell (cells clump,
fact of life) plate counts are
reported as the number of
If the concentration of bacteria is too great
Colony-Forming Units (CFU)
the colonies will grow into each other and
instead of the number of the plate will be uncountable.
cells.

To insure a countable plate a series of dilutions should be plated. The serial

dilutions should give at least one countable plate in the series (25-250 or 30-
300, depending on preference of the individual lab).

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Reference
 Hogg, Stuart (2013). Essential Microbiology, 2nd
ed., Wiley-Blackwell.

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