Burton’s Microbiology
for the Health Sciences
Section IV.
Controlling the Growth of
Microbes
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Burton’s Microbiology
for the Health Sciences
Chapter 8.
Controlling Microbial Growth
In Vitro
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Chapter 8 Outline
• Introduction
• Factors That Affect Microbial Growth
• Encouraging the Growth of Microbes In Vitro
• Inhibiting the Growth of Microbes In Vitro
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Microbiology - Chapter 8 & 9
Microbial Growth – Bacteria reproduce by “binary
fission”, a cell divides into two, two to four, four to eight,
etc.
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Microbiology – Chapter 8 & 9
• Cell division can occur quite rapidly depending on
nutrient levels, temperature, etc.
• E. coli can divide every 20 minutes (time to double –
generation time) in nutrient media at 37 degrees C.
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Microbiology - Chapter8 & 9
Microbial Growth is affected by two major
factors:
Environmental:
• temperature,
• pH,
Chemical: Proper concentrations of C, H, O,
N, P, S, some trace elements, and some
organic cofactors
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Factors That Affect Microbial Growth
• Availability of nutrients
– All living organisms require nutrients to sustain life.
– Nutrients are energy sources. Organisms obtain
energy by breaking chemical bonds.
• Moisture
– Water is essential for life. It is needed to carry out
normal metabolic processes.
– Certain microbial stages (e.g., bacterial endospores
and protozoal cysts) can survive a drying process
(desiccation).
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Factors That Affect Microbial Growth
(cont.)
• Temperature
– Every organism has an optimum growth temperature.
– Thermophiles are microorganisms that grow best at
high temperatures.
– Mesophiles are microbes that grow best at moderate
temperatures (e.g., 37°C, human body temperature).
– Psychrophiles prefer cold temperatures (like deep
ocean water).
– Psychroduric organisms prefer warm temperatures
but can endure very cold or even freezing
temperatures.
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Factors That Affect Microbial Growth
• pH
– pH refers to the acidity or alkalinity of a solution.
– Most microorganisms prefer a neutral or slightly
alkaline growth medium (pH 7.0–7.4).
– Acidophiles prefer a pH of 2 to 5
– Alkaliphiles prefer a pH of >8.5
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Encouraging the
Growth of Microbes
In Vitro
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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).
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Binary Fission of Staphylococci
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Culturing Bacteria in the Laboratory
Culture Media
• Media are used in microbiology laboratories to culture
(i.e., grow) bacteria; media prepared in the laboratories
are referred to as artificial media or synthetic media.
• A chemically defined medium is one in which all
ingredients are known.
• Culture media can be liquid or solid.
• An enriched medium is a broth or solid containing a rich
supply of special nutrients that promote the growth of
fastidious organisms, for example, chocolate agar.
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Examples of Solid and Liquid Culture Media
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Bacterial Colonies
Bacterial colonies on
S. aureus on mannitol
MacConkey agar (a
salt agar (a selective
selective and differential
and differential medium)
medium)
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Culturing Bacteria in the Laboratory
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.
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Inhibiting the Growth of
Microbes In Vitro
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Definition of Terms
• Sterilization is the complete destruction of all microbes,
including cells, spores, and viruses.
– Accomplished by dry heat, autoclaving (steam under
pressure), gas, various chemicals, and certain types
of radiation.
• Disinfection is the destruction or removal of pathogens
from nonliving objects by physical or chemical methods;
pasteurization is an example of a disinfection technique.
– Disinfectants are chemical substances that eliminate
pathogens on inanimate objects.
– Antiseptics are solutions used to disinfect skin and
other living tissues.
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Definition of Terms, cont.
• The suffix -cide or -cidal refers to “killing.”
• Germicidal agents, biocidal agents, and microbicidal
agents are chemicals that kill microbes.
• Bactericidal agents are chemicals that specifically kill
bacteria, but not necessarily bacterial endospores.
– Sporicidal agents kill bacterial endospores.
– Fungicidal agents kill fungi, including fungal spores.
– Algicidal agents kill algae.
– Viricidal agents destroy viruses.
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Definition of Terms (cont.)
• A microbistatic agent is a drug or chemical that inhibits
growth and reproduction of microbes.
• A bacteriostatic agent is one that specifically inhibits
the metabolism and reproduction of bacteria.
• Sepsis refers to the presence of pathogens in blood or
tissues, whereas asepsis means the absence of
pathogens.
• Antisepsis is the prevention of infection.
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Using Physical Methods to Inhibit
Microbial Growth
• Heat
– Two factors⎯temperature and time⎯determine the
effectiveness of heat for sterilization.
• Types of heat
– Dry heat (e.g., oven, electrical incinerator, or flame)
– Moist heat (boiling or use of an autoclave)
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Using Physical Methods to Inhibit
Microbial Growth, cont.
• The autoclave
– A large metal pressure
cooker that uses steam
under pressure to
completely destroy all
microbial life.
– Increased pressure
raises the temperature
above the temperature
of boiling water (above
100°C) and forces
steam into materials
being sterilized.
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Using Physical Methods to Inhibit
Microbial Growth, cont.
• Cold⎯most microorganisms are not killed, but their
metabolic activities are slowed.
• Desiccation⎯many dried microorganisms remain viable,
but they cannot reproduce.
• Radiation⎯an ultra-violet (UV) lamp is useful for reducing
the number of microbes in the air.
• Ultrasonic waves⎯used in hospitals and medical and
dental clinics to clean equipment.
• Filters⎯used to separate cells/microbes from liquids or
gases.
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Using Chemical Agents to Inhibit
Microbial Growth (cont.)
Characteristics of an ideal chemical antimicrobial
agent:
• Should have a broad • Soluble in water and easy to
antimicrobial spectrum apply
• Fast acting • Inexpensive and easy to
prepare
• Not affected by the presence
of organic matter • Stable as both a concentrate
and a working solution
• Nontoxic to human tissues
and noncorrosive • Odorless
• Should leave a residual
antimicrobial film on surface
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Using Chemical Agents to Inhibit
Microbial Growth (cont.)
• Antiseptics
– May be used safely on human tissues.
– Reduce the number of organisms on the surface of
the skin; do not penetrate pores and hair follicles.
• Antiseptic soaps and scrubbing are used by
healthcare personnel to remove organisms
lodged in pores or folds of the skin.
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Inhibiting the Growth of Pathogens in Our
Kitchens (from the CD-ROM)
• Many foods brought into our kitchens are contaminated
with pathogens (e.g., E. coli O157:H7, Salmonella, and
Campylobacter spp. on poultry and ground beef).
• Problems arise when handling foods before cooking.
• Remain aware of pathogens when preparing foods.
• Wash hands frequently.
• Thoroughly clean plates and countertops that have had
poultry or meat on them with hot soapy water.
• The use of antibacterial kitchen sprays is controversial.
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