Chapter 13

Control of Microbial
Growth
 The Terminology of Microbial Control
• _________refers to microbial contamination
• Asepsis is the absence of significant
contamination
• Aseptic surgery techniques prevent microbial
_______________________of wounds
 The Terminology of Microbial Control
• Sterilization: removing ______microbial life
• Commercial sterilization: killing C. botulinum
endospores
• Disinfection: _____________pathogens from
fomites
• Antisepsis: removing pathogens from living
tissue
 ____________ Biosafety Levels

• BSL2 Includes Clostridium difficle, Borrelia

burgdorferi; hepatitis, mumps,__________, and
measles viruses
 The Terminology of Microbial Control
• Degerming: removing microbes from a
___________________area
• Sanitization: lowering microbial counts on
eating utensils
• Biocide/germicide: killing microbes
• Bacteriostasis: inhibiting, ___ killing, microbes
Table 7.2 Microbial Exponential Death Rate: An Example
Understanding the Microbial _____________Curve.
Plotting the typical microbial death curve logarithmically
(______________) results in a straight line.

One log decrease =

Arithmetic number of surviving cells

90% of population
log10 of number of surviving cells
killed

Time (min)

(a) The decimal reduction time (D-value) is the time it takes to kill
90% of the population (a 1-log decrease in the total population)
when exposed to a specific microbial control protocol, as indicated
by the purple bracket.
 Heat
• Thermal death point (TDP): _______temperature
at which all cells in a culture are killed in 10 min
• Thermal death time (TDT): _____during which all
cells in a culture are killed___________________
• Decimal Reduction Time (DRT): time (minutes)
to kill __% of a population at a given temperature
 Effectiveness of Treatment
• Depends on:
– ___________________of microbes
– Environment (organic matter, temperature, biofilms)
– _____________________of exposure
– Microbial characteristics
An_______________________.
Exhaust valve Operating valve
(removes steam Steam to Safety (controls steam from
after sterilization) chamber valve Pressure gauge jacket to chamber)

Steam Door

Steam
chamber

Air

Perforated shelf Sediment

screen
Steam jacket Thermometer

Automatic ejector valve Pressure regulator

(thermostatically controlled; for steam supply
closes on contact with
pure steam when air is
To waste line exhausted) Steam supply
 Pasteurization
• _________spoilage organisms and pathogens
 Dry Heat Sterilization

• Kills by __________
– Dry heat
– Flaming
– Incineration
– Hot-air sterilization
Hot-Air Autoclave

Equivalent Treatments 170˚C, 2 hr 121˚C, 15 min

 Filtration
• ___________ removes microbes >0.3 µm
• Membrane filtration removes microbes >0.22
µm
The roles of high-efficiency particulate air (____) filters in biological safety cabinets.
Outside

Safety glass
Exhaust HEPA viewscreen
filter

Blower

Supply HEPA
filter

Light

High-velocity
air barrier
 Radiation
• Ionizing radiation (X rays, ________rays, electron beams)
– Ionizes water to release OH•
– Damages _____
• Nonionizing radiation (UV, 260 nm)
– Damages ____________
• Microwaves kill by heat; not especially antimicrobial
 Principles of _______Disinfection
• Concentration of disinfectant
• Organic matter
• pH
• Time
 Use-Dilution Test
• Metal _____ dipped in test bacteria are dried
• Dried cultures are placed in disinfectant for 10
min
at 20°C
• Rings are transferred to culture media to
determine whether bacteria _______
treatment
 Chemical Methods of Microbial Control
• In-Use Test
– 1 ml sample of _____ disinfectant in 9 ml sterile broth
– 0.2 ml inoculated onto two plates.
– Incubated
• __ or more colonies indicate disinfectant not effective
Evaluation of disinfectants by the ____-diffusion method.

Zone of inhibition

Chlorine Chlorine Chlorine

O-phenylphenol O-phenylphenol
Hexachlorophene O-phenylphenol
Hexachlorophene Hexachlorophene

Quat Quat
Quat

Staphylococcus aureus Escherichia coli Pseudomonas aeruginosa

(gram-positive) (gram-negative) (gram-negative)
 Phenol and Phenolics
• Disrupt plasma membranes

_________
 Chemical Methods of Microbial Control
• Methods for Evaluating Disinfectants and Antiseptics
– __________________ coefficient
• Evaluates efficacy of disinfectants and antiseptics
• Compares to phenol an agent's ability to control
microbes
• Greater than 1.0 indicates agent is _____ effective than
phenol
• Has been replaced by newer methods
 Halogens

• Iodine
– Tinctures: in ______________ alcohol
– Iodophors: in organic molecules (Betadine)
– Alter protein synthesis and membranes
• Chlorine
– Bleach: hypochlorous acid (HOCl)
– Chloramine: chlorine + _____________
– Oxidizing agents
 Alcohols
• Ethanol, isopropanol
– _________ proteins, dissolve lipids
– Require water
 Heavy Metals
• Ag, Hg, and Cu
– Silver nitrate may be used to prevent gonorrheal
ophthalmia neonatorum
– Silver sulfadiazine used as a topical cream on burns
– Copper sulfate is an ____________________
• Oligodynamic action
– Denature proteins
 Surface-Active Agents, or _______

Soap Degerming

Acid-anionic detergents Sanitizing

Quaternary ammonium compounds Bactericidal, denature proteins,

(cationic detergents) disrupt plasma membrane
The ammonium _____ and a quaternary ammonium compound, benzalkonium chloride
(Zephiran).

Ammonium ion Benzalkonium chloride

 Chemical Food Preservatives
• Organic ________________
– Inhibit metabolism
– Sorbic acid, benzoic acid, and calcium propionate
– Control molds and bacteria in foods and cosmetics
• Nitrite prevents endospore germination
• Antibiotics
– _____ and natamycin prevent spoilage of cheese
 Aldehydes
• Inactivate __________ by cross-linking with
functional groups (–NH2, –OH, –COOH, –SH)
• _____: medical equipment
– Glutaraldehyde, formaldehyde, and ortho-
phthalaldehyde
 Gaseous Sterilants
• Denature _________________s
• Use: heat-sensitive material
– Ethylene _____
 Peroxygens
• Oxidizing agents
• Use: contaminated surfaces
– O3, ______, peracetic acid
________________ order of resistance of microorganisms to chemical biocides.
The Effectiveness of Chemical Antimicrobials against Endospores and __________