Chapter08Controlling The Growth of Microbes

• Factors that Affect Microbial Growth
•Encouraging the Growth of Microbes

in Vitro
•Inhibiting the Growth of Microbes in
Vitro
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Factors That Affect Microbial Growth

Factors That Affect Microbial Growth, cont.






Changes in Osmotic Pressure

Factors That Affect Microbial Growth cont.

Encouraging the
Growth of Microbes in
Vitro

Culturing Bacteria in the Laboratory
Bacterial Growth
• Think of bacterial growth as an increase in the number of
organisms rather than an increase in their size.
• Bacteria divide by binary fission (one cell divides to
become two cells) when they reach their optimum size.
• Binary fission continues through many generations until a
colony is produced on solid culture medium.
• Binary fission continues for as long as there is a sufficient
supply of nutrients, water, and space.
• The time it takes for one cell to become two cells is called
the generation time (e.g., E. coli = 20 minutes).

Binary fission of
staphylococci.

Culture Media
Phenylethyl alcohol (PEA) agar allows growth of gram-

positive organisms, particularly cocci, while inhibiting most
gram-negative bacteria andKluwer
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Health | Lippincott Williams & Wilkins
Culture Media, cont.

A Thioglycollate (THIO)
Broth Tube

Bacterial colonies on S. aureus on mannitol-salt
MacConkey agar (a agar (a selective &
selective & differential differential medium)
medium)

Colonies of a β-hemolytic Streptococcus species on a blood agar
plate (in this case, the blood agar is both enriched and differential)

Inoculation of Culture Media
• Culture media are
inoculated with clinical
specimens (i.e.,
specimens collected
from patients with a
suspected infectious
disease).
• Inoculation involves
adding a portion of a
specimen to the
medium.
• Inoculation is
accomplished using a
sterile inoculating loop.
Importance of Using “Aseptic Technique”

Incubation

Bacterial Population Counts

Measurement of Bacterial Growth
• Direct method
– cell counts
• either via a microscope or cell counter
• Indirect methods
– colony counts (viable cell counts)
– Spectrophotometry (turbidity)
– Weight (dry vs wet)
– ATP determination

Bacterial Population Growth Curve

A population growth curve of living
organisms.
Stationary phase
Death phase
Logarithmic growth phase
Lag phase
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| Lippincott Wolters
Williams Kluwer Health | Lippincott Williams & Wilkins
& Wilkins
Lag Phase
 Occurs when organisms are
transferred to new medium
 Little increase in cell number •
 BacteriaLagare
Phasetransporting nutrients
inside cell from the new medium •
 Cells are preparing for replication
and division •
 Individual cells increase in size
 growth is generally unbalanced
Exponential (or log) Phase
• Bacterial cell division begins
– proceeds as a geometric progression
– cell numbers increase as an
exponential function of time
• Growth is unrestricted, but balanced
– the concentration of all
macromolecules within the cells are
increasing at the same rate
Stationary Phase
• No net increase in cell numbers

– growth rate = death rate
• Brought about by exhaustion of
nutrients, waste product buildup or
changes in physical conditions •
Metabolic rate decreases
– Feedback mechanisms regulate
enzymes involved in key metabolic steps
Stationary Phase
• Cells are more resistant to environmental
stresses
• Significant physiological changes can occur
between cells in log phase and those in
stationary phase
– eg. Arthobacter cells change from rod-shaped
cells (log phase) to cocci (stationary phase)
• Growth is unbalanced
– various cellular components are synthesised
disproportionately
Death Phase
• Decline in cell numbers
• Brought about when toxins or waste
products reach a threshold
concentration
• Bacterial growth is restricted and
unbalanced
– cells cannot obtain all requirements
for growth or replication
Growth of Bacterial Cultures
• In nature
– conditions cannot be controlled
– many species co-exist
– changes in conditions may cause population
shifts as conditions favor certain members of
the population over others
• In the lab
– conditions can be controlled
– established to favor a particular organism
_ important in industrial processes for the
accumulation of desired metabolic products
Batch Cultures
• produce bacterial growth curves
just discussed
• inoculation of fresh media with
bacteria
• nutrients are expended
• metabolic products accumulate
• closed environment
– eg. Inoculating 100 mL of a rich
media in a 1 L flask with E. coli
Continuous Culture
• Fresh medium replaces spent medium

– continuous replenishing of nutrients and
removal of waste products
– permits continuous growth of culture
• Continuous culturing can be controlled by …
– Turbidostat
• monitors turbidity and cycles media as
required
– Chemostat
• constant flow rate continuously cycles
media
• keeps cells in log phase
Synchronous Culture
Synchronous growth
– all cells divide at the same time
– can be achieved by altering
environmental conditions
• repeatedly changing the
temperature
• adding fresh media to cells
entering stationary phase
– can only be maintained for a few
generationsCopyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins
Culturing Obligate Intracellular Pathogens in
the Laboratory

Culturing Fungi in the Laboratory

Culturing Protozoa in the Laboratory

Inhibiting the Growth of Microbes in-
Vitro

Definition of Terms

Definition of Terms, cont.

Definition of Terms (cont.)

Using Physical Methods to Inhibit Microbial
Growth

Dry Heat Sterilization
Using a Bunsen Using an electrical

burner flame heating device

Growth, cont.

A large, built-in autoclave.

Pressure-sensitive autoclave tape showing dark stripes
after sterilization.

Biological Indicators for Monitoring the
Effectiveness of Steam Sterilization

Growth, cont.

Using Chemical Agents to Inhibit Microbial
Growth

Growth, cont.
Characteristics of an ideal chemical antimicrobial
agent:
• Should have a broad • Soluble in water and
antimicrobial spectrum easy to apply
• Fast acting • Inexpensive and easy to
prepare
• Not affected by the
presence of organic matter • Stable as both a
concentrate and a
• Nontoxic to human tissues working solution
and noncorrosive
• Odorless
• Should leave a residual
antimicrobial film on
surface
Growth (cont.)

Inhibiting the Growth of Pathogens in Our
Kitchens

Chapter08Controlling The Growth of Microbes

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• Factors that Affect Microbial Growth

•Encouraging the Growth of Microbes

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Phenylethyl alcohol (PEA) agar allows growth of gram-

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Logarithmic growth phase

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• No net increase in cell numbers

• Fresh medium replaces spent medium

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Using a Bunsen Using an electrical

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Copyright © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

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