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Microbial Control
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Basic Principles of
Microbial Control
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Figure 9.2
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Figure 9.3
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• Heat-Related Methods
▫ Effects of high temperatures
Denature proteins
Interfere with integrity of cytoplasmic membrane
and cell wall
Disrupt structure and function of nucleic acids
▫ Thermal death point
Lowest temperature that kills all cells in broth in 10
min
▫ Thermal death time
Time to sterilize volume of liquid at set temperature
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• Heat-Related Methods
▫ Moist heat
Used to disinfect (kill organisms and remove spores),
sanitize (kill organisms but not necessarily their spores),
and sterilize (kill all organisms and spores)
Denatures proteins and destroys cytoplasmic membranes
More effective than dry heat
Methods of microbial control using moist heat
Boiling
Autoclaving
Pasteurization
Ultrahigh-temperature sterilization
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• Heat-Related Methods
▫ Moist heat
Boiling
Kills vegetative cells of bacteria and fungi, protozoan
trophozoites, and most viruses
Boiling time is critical
▫ Different elevations require different boiling times
Endospores, protozoan cysts, and some viruses can
survive boiling
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• Heat-Related Methods
▫ Moist heat
Autoclaving
Pressure applied to boiling water prevents steam from
escaping
Boiling temperature increases as pressure increases
Autoclave conditions – 121ºC, 15 psi, 15 min
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Figure 9.5
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Sterility indicator
Figure 9.7
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• Heat-Related Methods
▫ Moist heat
Pasteurization
Used for milk, ice cream, yogurt, and fruit juices
Not sterilization
▫ Heat-tolerant microbes survive
Pasteurization of milk
▫ Batch method
▫ Flash pasteurization (High temp, short time)
▫ Ultrahigh-temperature pasteurization (very high temp,
very short time)
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Pasteurization of milk
Batch method
• The batch method uses a vat pasteurizer which consists
of a jacketed vat surrounded by either circulating water,
steam or heating coils of water or steam.
Pasteurization of milk
Flash method
• High Temperature Short Time (HTST)
• Milk is heated to 72°C (161.6°F) for at least 15 seconds.
• Used for perishable beverages like fruit and vegetable
juices, beer, and some dairy products. Compared to
other pasteurization processes, it maintains color and
flavor better.
• It is done prior to filling into containers in order to kill
spoilage microorganisms, to make the products safer and
extend their shelf life. Flash pasteurization must be used
in conjunction with sterile fill technology.
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Pasteurization of milk
Ultrahigh-temperature method
• Heating for 1-2 seconds at a temperature exceeding
135°C (275°F), which is the temperature required to kill
spores in milk.
• The most common UHT product is milk, but the process
is also used for fruit juices, cream, soy milk, yogurt,
wine, soups, and stews.
• Can cause browning and change the taste and smell of
dairy products.
• UHT canned milk has a typical shelf life of six to nine
months, until opened.
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• Heat-Related Methods
▫ Moist heat
Ultrahigh-temperature sterilization
140ºC for 1 sec, then rapid cooling
Treated liquids can be stored at room temperature
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• Heat-Related Methods
▫ Dry heat
Used for materials that cannot be sterilized with
moist heat
Denatures proteins and oxidizes metabolic and
structural chemicals
Requires higher temperatures for longer time than
moist heat
Incineration is ultimate means of sterilization
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Figure 9.8
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Figure 9.9
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High-Efficiency Particulate
Arresting (HEPA) air filters are
used in medical facilities,
automobiles, aircraft, and
homes. The filter must remove
99.97% of all particles greater
than 0.3 micrometer from the
air that passes through.
Figure 9.10
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• Osmotic Pressure
▫ High concentrations of salt or sugar in foods to
inhibit growth
▫ Cells in hypertonic solution of salt or sugar lose
water
▫ Fungi have greater ability than bacteria to survive
hypertonic environments
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• Radiation
▫ Ionizing radiation
Wavelengths shorter than 1 nm
Electron beams, gamma rays
Ejects electrons from atoms to create ions
Ions disrupt hydrogen bonding, cause oxidation, and
create hydroxide ions
Hydroxide ions denature other molecules (DNA)
Electron beams – effective at killing but do not
penetrate well
Gamma rays – penetrate well but require hours to kill
microbes
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Figure 9.11
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• Radiation
▫ Nonionizing radiation
Wavelengths greater than 1 nm
Excites electrons, causing them to make new covalent
bonds
Affects 3-D structure of proteins and nucleic acids
UV light causes pyrimidine dimers in DNA
UV light does not penetrate well
Suitable for disinfecting air, transparent fluids, and
surfaces of objects
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• Biosafety Levels
▫ Four levels of safety in labs dealing with
pathogens
Biosafety Level 1 (BSL-1)
Handling pathogens that do not cause disease in
healthy humans
Biosafety Level 2 (BSL-2)
Handling of moderately hazardous agents
Biosafety Level 3 (BSL-3)
Handling of microbes in safety cabinets
Biosafety Level 4 (BSL-4)
Handling of microbes that cause severe or fatal disease
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Figure 9.12
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• Alcohols
▫ Intermediate-level disinfectants
▫ Denature proteins and disrupt cytoplasmic
membranes
▫ More effective than soap in removing bacteria
from hands
▫ Swabbing of skin with 70% ethanol prior to
injection
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• Halogens
▫ Intermediate-level antimicrobial chemicals
▫ Believed to damage enzymes via oxidation or by
denaturation
▫ Widely used in numerous applications
Iodine tablets, iodophores, chlorine treatment,
bleach, chloramines, and bromine disinfection
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Figure 9.14
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• Oxidizing Agents
▫ Peroxides, ozone, and peracetic acid
▫ Kill by oxidation of microbial enzymes
▫ High-level disinfectants and antiseptics
▫ Hydrogen peroxide (H2O2) can disinfect and
sterilize surfaces
Not useful for treating open wounds due to catalase
activity: the tissues convert it into H20 and 0ygen
bubbles.
▫ Ozone treatment of drinking water
▫ Peracetic acid is an effective sporocide used to
sterilize equipment
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• Surfactants
▫ “Surface active” chemicals
Reduce surface tension of solvents
▫ Soaps and detergents
Soaps have hydrophilic and hydrophobic ends
Good degerming agents but not antimicrobial
Detergents are positively charged organic surfactants
▫ Quats (Quaternary ammonium cations)
Low-level disinfectants; disrupts cell membranes
Ideal for many medical and industrial applications
Good against fungi, amoeba, and enveloped viruses,
but not endospores, Mycobacterium tuberculosis and
non-enveloped viruses.
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• Heavy Metals
▫ Heavy-metal ions denature proteins
▫ Low-level bacteriostatic and fungistatic agents
▫ 1% silver nitrate to prevent blindness caused by N.
gonorrhoeae
▫ Thimerosal used to preserve vaccines
▫ Copper inhibits algal growth
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• Aldehydes
▫ Compounds containing terminal –CHO groups
▫ Cross-link functional groups to denature proteins
and inactivate nucleic acids
▫ Glutaraldehyde disinfects and sterilizes
▫ Formalin used in embalming and disinfection of
rooms and instruments
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• Gaseous Agents
▫ Microbicidal and sporicidal gases used in closed
chambers to sterilize items
▫ Denature proteins and DNA by cross-linking
functional groups
▫ Used in hospitals and dental offices
▫ Disadvantages
Can be hazardous to people
Often highly explosive
Extremely poisonous
Potentially carcinogenic
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• Enzymes
▫ Antimicrobial enzymes act against microorganisms
▫ Human tears contain lysozyme
Digests peptidoglycan cell wall of bacteria
▫ Enzymes to control microbes in the environment
Lysozyme used to reduce the number of bacteria in
cheese
Prionzyme can remove prions on medical
instruments
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• Antimicrobials
▫ Antibiotics, semi-synthetic, and synthetic
chemicals
▫ Typically used for treatment of disease
▫ Some used for antimicrobial control outside the
body
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Antimicrobial Agents
• Semi-synthetics
▫ Chemically altered antibiotics that are more
effective than naturally occurring ones
• Synthetics
▫ Antimicrobials that are completely synthesized in
a lab
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Mechanisms of Antimicrobial
Action
• Inhibition of bacterial wall synthesis
• Disruption of existing cytoplasmic membranes
• Inhibition of Protein Synthesis
• Inhibition of Nucleic Acid Synthesis
• Inhibition of Metabolic Pathways
• Prevention of Virus Attachment
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Figure 10.4
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Nucleotides
Figure 10.7
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Acyclovir
• Acyclovir is used to decrease pain and speed the healing of herpes sores or
blisters in people who have varicella (chickenpox), herpes zoster (shingles;
a rash that can occur in people who have had chickenpox in the past), and
first-time or repeat outbreaks of genital herpes (a herpes virus infection
that causes sores to form around the genitals and rectum from time to
time).
• Acyclovir is also sometimes used to prevent outbreaks of herpes sores in
people who are infected with the virus.
• Acyclovir disrupts nucleic acid function. It works by stopping the spread of
the herpes virus in the body. Acyclovir will not cure herpes or protect others
from catching it.
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Nucleotides
Figure 10.7
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Figure 10.8
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Figure 10.9
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Figure 10.10
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Figure 10.12
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Effect of route of
administration on
chemotherapeutic
agent
Figure 10.13
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Figure 10.14
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Figure 10.15
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Figure 10.16
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• Retarding Resistance
▫ Maintain high concentration of drug in patient for
sufficient time
Kills all sensitive cells and inhibits others so immune
system can destroy
▫ Use antimicrobial agents in combination
Synergism vs. antagonism
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Figure 10.17
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Vaccination
• Vaccine – use the immune system to protect
against infectious disease
• Types of vaccines
▫ attenuated (weakened) microbe; virulence factors are
removed
▫ heat-killed / chemically killed microbe
▫ toxoids
• Passive versus Adaptive vaccination
▫ passive – immune system products from another
mothers milk (presence of IgA)
gamma-globulin (anti-bee venom, anti-hepatitis A, etc)
▫ active – stimulate individuals immune system to produce
memory cells
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U.S. cases
against
diseases for
which there
are vaccines.
• http://news.yahoo.com/why-cellphone-more-
bacteria-toilet-seat-124147769.html
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