Microbial Growth

Basic Microbiology
Department of Biology
FMIPA-IPB

Microbial Growth
Because individual cells grow larger only
to divide into new individuals, microbial
growth is defined not in terms of cell size
but as the increase in the number of cells,
which occurs by cell division."

Binary Fission

Exponential Growth

Batch Culture

Standard Bacterial Growth Curve

Log Phase

Log Phase

Growth in
Continuous Culture
A continuous culture is an open system in which fresh
media is continuously added to the culture at a constant
rate, and old broth is removed at the same rate.
This method is accomplished in a device called a
chemostat.
Typically, the concentration of cells will reach an
equilibrium level that remains constant as long as the
nutrient feed is maintained.

Kultur Kemostat

Chemostat System

Standard Bacterial Growth Curve

During lag phase, cells are recovering from a period of no

growth and are making macromolecules in preparation for
growth
During log phase cultures are growing maximally
Stationary phase occurs when nutrients are depleted and wastes
accumulate (Growth rate = death rate)
During death phase death rate is greater than growth rate
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Decline (Death) Phase

Methods used to measure

microbial growth
Count colonies on plate or filter (counts
live cells)
Microscopic counts
Flow cytometry
Turbitity
Most Probable Number Method (MPN)
14

Viable counts

Each colony on plate or filter arises from single live cell

Only counting live cells
15

Microscopic counts

Need a microscope, special slides, high power

objective lens
Typically only counting total microbe numbers, but
differential counts can also be done

16

Flow cytometer (FACS)

Florescent antibodies are
added to culture and cells in
small droplets are sent
through a detector single file
Computers count and
characterize cells as they
pass, and deflect cells with
desired characteristics
Can count and keep live cells

flow-cytometry.de/img/fcm.gif

17

Turbidity
Cells act like large particles
that scatter visible light
A spectrophotometer sends a
beam of visible light through
a culture and measures how
much light is scattered
Scales read in either
absorbance or % transmission
Measures both live and dead
cells
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Most Probable Number Method

Most Probable Number Method

Most Probable Number Calculations

(based on 5 tubes/dilution)
dilutions (100 = 1 ml culure)
100

10-1

10-2

10-3

10-4

MPN/ml

33

33

40

40

5400

0.20

4.7

pH

Neutrophiles

Temperature

Temperature

Thermophiles

A large channel draining from a hot pool,

containing carotenoid-rich microorganisms. The
temperature of this channel in the foreground is
about 60 oC. Layers of white-coloured limestone
(forming a rock deposit known as travertine) can
also be seen. Note the footprints of buffalo in the
foreground. These animals often seek the warmth
of thermal areas in the winter months.

limestone terraces formed by precipitation

from calcium-rich water flowing from a raised
hot pool. Pink, green and brown-coloured
microorganisms occupy the thermal gradients
in the flowing water (60-100oC)

Thermophiles

Deep-Sea Vents

Classification of organisms
based on O2 utilization
Utilization of O2 during metabolism yields toxic
by-products including O2-, singlet oxygen (1O2)
and/or H2O2.
Toxic O2 products can be converted to harmless
substances if the organism has catalase (or
peroxidase) and superoxide dismutase (SOD)
SOD converts O2- into H2O2 and O2
Catalase breaks down H2O2 into H2O and O2
Any organism that can live in or requires O2 has
SOD and catalase (peroxidase)
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Classification of organisms
based on O2 utilization

Obligate (strict) aerobes require O2 in order to grow

Obligate (strict) anaerobes cannot survive in O2
Facultative anaerobes grow better in O2
Aerotolerant organisms dont care about O2
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Microaerophiles require low levels of O2

Growing Obligate Anaerobes

Fluid Thioglycolate Medium

Peptones, Yeast Extracts, Glucose
Thioglycolate binds O2
Agar slows diffusion of O2
Resazurin is O2 indicator (red)

Growing Obligate Anaerobes

Anaerobic Jar & Chamber

Salt Tolerance

Media

Media

Media

Selective and Differential

Media
The concepts of Selective and
Differential media can be confusing,
particularly because very often media
are both Selective and Differential

Differential Media