Chapter 11: Physical and Chemical Control of Microbes

I. Joseph ______ aseptic

Lister started _________ techniques with medical
phenol
applications. By using carbolic acid (_______) -soaked rags and
instruments during and after surgery, gangrene and other infections
following surgery greatly diminished.
II. Terminology and Methods of Control
Sterilization
A. _____________means COMPLETE destruction of viruses and
microbes (including endospores) so that even if they are
placed
in a new growth medium, they will not revive or reproduce.
Disinfection
B. __________means to reduce the number of pathogens (including
viruses) until they are not a hazard, usually involving the use of
antimicrobial chemicals.
Decontamination
C. _______________
Sanitization refers to removing toxins.
D. ____________ refers to a substantially
reduced microbial population that meets
accepted health standards.
A clean appearance is expected!
E. Different situations warrant different levels of microbial
control.
1. daily life
handwashing with plain soap and water
Simple ____________
is considered to be the single most important step in
preventing the spread of many infectious diseases!
 2. hospitals
nosocomial
Danger of ___________ (hospital acquired) infections because of:
weakened
a. _________ condition of hospitalized patients
b. higher concentration of sick people withpathogenic
____________
microbes (*many resistant forms!!)
invasive
c. _______ procedures (such as)

carriers
d. many health care workers are ______
aseptic
e. lack of _______ care (handwashing
between patients, using gloves, etc.)
3. microbiology/research/hospital laboratories
aseptic techniques
must use ________
clean .
a. Work surfaces should be ______
sterile .
b. All media and instruments must be ______
cultures must be properly disposed of.
c. Used ________
III. Selection of an antimicrobial procedure depends on many factors
microbe the extent of ____________,
such as the type of _______, contamination
environmental conditions, and potential risk of _________.
____________ infection
A. types of resistant microbes
endospores
1. Bacillus and Clostridium can make ___________.
2. Mycobacterium has ______waxy cell walls.
Pseudomonas is capable of metabolizing unusual
3. ____________
substances for food. (Like disinfectants!)
B. the extent of contamination (size of the microbial population)
90
1. ‘Industry standard’ requires that ____% of the population is
killed with every __ minutes 2 of exposure to the treatment
a. 100 microbes  10 microbes  1 microbe in __ 4 minutes
b. 1010 microbes        would take ___ 20 minutes
washing scrubbing first helps reduce the population
SO, ________/_________
before disinfection or sterilization.
C. environmental conditions
temperature
1. _____________ ( heat  chemical action)
pH
2. _____
3. ____, saliva _______,
dirt _______, blood feces can all block chemical action
______
D. Potential risk of infection
Critical items come into direct contact with body tissues.
1. _______

Semicritical
2. ____________ items come into contact with mucous membranes,
but do not penetrate body tissues.
 Noncritical items only touch keratinized skin surfaces.
3. ____________

IV. Methods of Physical Control

denaturing cell proteins /enzymes. It is the
Heat works by_________
A. ______
most common control method because it is fast, reliable,
inexpensive & nontoxic.
Moist heat
1. ______
Boiling 100°C/10 minutes (kills most microbes &
a. _______
endospores
inactivates most viruses, but does not destroy __________).
 Pasteurizatio
b. ____________: a brief heat treatment followed by rapid cooling.
n(Kills pathogens and reduces the number of spoilage organisms in
milk, juices, wine, beer: Does not sterilize!)
(1). LTLT (Low Temperature Long Term) 63°C/30 minutes
*(2). HTST (High Temperature Short Term) 72°C/15 seconds

Autoclave (steam under pressure)

c. __________
(1). 15-20 psi/15-20 minutes/121°C
Sterilizes equipment, media, etc.
(2). ________
(3). used in canning procedures to destroy
endospores
Clostridium botulinum __________!
2. ___
Dry heat sterilizes.
moisture is
a. Hot air ovens (160-170°C/2-3 hours) used when ________
undesirable.
Incineration (burning)
b. ____________
furnaces incinerators
(1). _________/___________used to destroy disposable items,
soiled dressings, tissue specimens etc. @ 800°C to 6500°C
c. The hottest part of a Bunsen burner flame reaches 1,870°C for
flaming
______ during lab.

Microbiology
is Fun!
B. Radiation (waves having energy but no mass) causes lethal
changes in DNA, denatures proteins, but doesn’t reliably destroy
endospores)!
ultraviolet
1. Nonionizing rays = ________
radiation
a. can be used to reduce the number
of organisms in air and on clean
surfaces
b. of limited use, cannot penetrate
materials like cloth, glass, paper
X-rays or _____________
2. Ionizing rays = ________ Gamma rays
sterilize
a. can be used to __________ items that are
heat or chemical sensitive, such as plastics
b. more effective, penetrates liquids and most
solids (used to treat Washington DC mail)
c. In the US, radiation is approved to treat
pork to preventtrichinosis
___________, to treat beef
forE. coli
________ contamination and used to
Salmonella
treat chicken for _________ contamination.
 3. microwaves
heat they
a. do not affect microbes directly, but may kill by _____
generate
uneven
b. drawback is that microwave heating is ________
C. Filtration (may be used for air, some heat sensitive materials such
as serum, vaccines, drugs, IV fluidsbeer/wine)
High-Efficiency
1. _____ ________Particulate
________ Air
____ (HEPA) filters remove
airborne contaminants; used in operating rooma, for people with
allergies, etc.
solids
2. In fluid filtration, _______ liquids
are separated from ________ by
passing through _______filters
with extremely fine pores
a. Mechanical force or vacuum suction helps fluid through the filter
b. does not sterilize unless pore size is small enough to trap
everything (smaller pores,  cost)
 Chemical Control (* for heat sensitive items, large
V. Methods of ________
surfaces)
Destructive actions include injury to the cell _________,
membrane
denaturation of proteins DNA
cell ________, inhibiting replication of
_____.

A. Disinfectants Vs Antiseptics
Disinfectants
1. _____________
Germicides are chemicals used on inanimate objects.
a. ___________ are chemicals that KILL/DESTROY germs.
Germistatic
(examples: fungicides, bactericides, viricides)
b. __________ refers to chemicals that do not kill, but
prevent the growth of microbes .
Antiseptics
(examples: bacteriostatic, fungistatic)
2. __________ are disinfectants nontoxic enough to be used on
skin. potency
B. Germicides are grouped according to their _______ (strength)
Sterilants
1. __________ destroy everything, including endospores
(for sterilizing scalpels, respiratory therapy equipment,
proctoscopes, plastic Petri dishes, endoscopes)
(ethylene oxide gas, hydrogen peroxide)
 High level disinfectants (do not reliably destroy endospores)
2. ____
(used for GI endoscopes) (iodine, phenol, chlorhexidine, heavy
metals such as silver nitrate)
Intermediate level disinfectants (will kill Mycobacterium, but do
3. ___________
not destroy all viruses or endospores, even with prolonged
exposure) (used for stethoscopes, electrodes, thermometers)
(alcohols: ethyl alcohol, isopropyl)
Low level disinfectants (will not kill Mycobacterium)
4. _____
(soaps, detergents)
 Phenol coefficient
C. ______ _________ (5% Phenol is the standard against which
chemical agents are tested and compared)
1. Each chemical is compared for the same length oftime
_____
on the sameorganism
_________ under identical
________ conditions
2. IF the chemical being tested requires a greaterconcentration
____________ time less
or a longer ______ than phenol, its efficiency is _____ than
phenol.

IF the chemical being tested requires a lower concentration

greater
or a shorter time than phenol, its efficiency is _______ than
phenol.

3. Ratio of: tested chemical activity

less phenol activity
_____ efficient than phenol
more
< 1 means
> 1 means _____ efficient than phenol
D. Selecting the Appropriate germicidal chemical
Toxicity (the benefit of disinfecting or sterilizing an item or
1. ________
surface must be weighed against the risks associated with the use
of that chemical) (hospital Vs home/office)
material being treated (metal, rubber,
2. compatibility with the ________
glass, plastic)
Residue may necessitate rinsing
3. ________
Cost and availability (bleach)
4. ______
Storage
5. _________ and stability (concentrates require less space and
store for long periods, but when diluted/mixed, often have limited
shelf life)
Environmental
6. _____________risk (disposal procedures needed)
VI. Methods used for Preservation (delaying spoilage) of Perishable
Products
A. Chemical
________ preservatives (both nonfood and food)
1. organicacids
______ lower pH (inactivates enzymes, inhibits
growth, but does not always destroy microbes)
nitrates
2. ________ and _______ inhibit germination of Clostridium
nitrites
botulinum endospores!
B. Low Temperatures
refrigerator
1. _____________
a. 0-10° C4(___° C average)
b. retards but does not prevent growth
freezer
2. ________
-20
a. ___° C
b. prevents growth but does not kill all organisms
osmotic pressure by adding _____
C. Increased _______ sugar
salt or ______;
causes water to leave the cell, killing it.
Desiccation (dehydration) of the material (natural [sun] or
D. ___________
artificial)
E. ____________
Lyophilization (freeze-drying)
rapidly frozen at temperatures well below 0°C
1. materials _______
moisture (lightweight)
2. vacuum while frozen to remove ________
3. biological cultures, medications, foods (expensive)
 Chap 12: Elements of Chemotherapy
I. Terminology
Chemotherapy = use of chemical agents to treat disease
A. ____________
Chemotherapeutic
B. _________________ agent (CTA) = chemical agent used for
treatment of disease (even cancer)
Antimicrobial
C. _____________ agent (AMA) = chemical agent used to treat
diseases caused by microbes
II. Antimicrobial Agents
A. Types of antimicrobial agents
Natural agents = metabolic products produced by
1. ________
certain groups of fungi and fungal-like bacteria that
are antibacterial in action
Synthetic
2. __________ agents = produced in the laboratory
Semi-synthetic
3. _______________ agents = derivatives of natural agents
altered in the laboratory by adding chemical groups to
improve effectiveness
B. Modes of action
1. interfere with microbe’s chemosynthesis by inhibiting ________ enzymes
2. Disruption/interference with
a. of an essential metabolite by _________
competitiveinhibition
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p. 77)
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid cross-
linkages of peptidoglycan) (p. 78)
c. by damaging the cell ___________ (Polymixin cleaves the
layers of the membrane like a knife) (p. 78)
d. by inhibiting ________________ at 70s ribosomes (p. 79)
(Erythromycin inhibits translocase, freezing the ribosome on
the mRNA.)
(Tetracycline blocks tRNA attachment to mRNA)
(Chloramphenicol inhibits transferase, preventing peptide
bond formation between amino acids.)
(Streptomycin causes a misreading of mRNA.)
e. by inhibiting nucleic acid (______ and/or ____) synthesis
(Antiviral: AZT inhibits reverse transcriptase.)
(Antibacterial: rifampin inhibits RNA polymerase.)
(Antifungal: griseofulvin inhibits RNA polymerase.)
 Folic acid

PABA
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
competitive
a. of an essential metabolite by _________ inhibition
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p.
77) wall
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid
cross-
linkages of peptidoglycan) (p. 78)
c. by damaging the cell ___________ (Polymixin cleaves the
layers of the membrane like a knife) (p. 78)
d. by inhibiting ________________ at 70s ribosomes (p. 79)
(Erythromycin inhibits translocase, freezing the ribosome on
the mRNA.)
(Tetracycline blocks tRNA attachment to mRNA)
(Chloramphenicol inhibits transferase, preventing peptide
bond formation between amino acids.)
(Streptomycin causes a misreading of mRNA.)
e. by inhibiting nucleic acid (______ and/or ____) synthesis
(Antiviral: AZT inhibits reverse transcriptase.)
(Antibacterial: rifampin inhibits RNA polymerase.)
(Antifungal: griseofulvin inhibits RNA polymerase.)
Glycan
“backbone”
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
competitive
a. of an essential metabolite by _________ inhibition
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p.
77) wall
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid
cross- membrane
linkages of peptidoglycan) (p. 78)
c. by damaging the cell ___________ (Polymixin cleaves the
layers of the membrane like a knife) (p. 78)
d. by inhibiting ________________ at 70s ribosomes (p. 79)
(Erythromycin inhibits translocase, freezing the ribosome on
the mRNA.)
(Tetracycline blocks tRNA attachment to mRNA)
(Chloramphenicol inhibits transferase, preventing peptide
bond formation between amino acids.)
(Streptomycin causes a misreading of mRNA.)
e. by inhibiting nucleic acid (______ and/or ____) synthesis
(Antiviral: AZT inhibits reverse transcriptase.)
(Antibacterial: rifampin inhibits RNA polymerase.)
(Antifungal: griseofulvin inhibits RNA polymerase.)
hydrophilic

amphipathic

hydrophobic
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
competitive
a. of an essential metabolite by _________ inhibition
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p.
77) wall
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid
cross- membrane
linkages of peptidoglycan) (p. 78)
c. by damagingprotein
the cellsynthesis
___________ (Polymixin cleaves the
layers of the membrane like a knife) (p. 78)
d. by inhibiting ________________ at 70s ribosomes (p. 79)
(Erythromycin inhibits translocase, freezing the ribosome on
the mRNA.)
(Tetracycline blocks tRNA attachment to mRNA)
(Chloramphenicol inhibits transferase, preventing peptide
bond formation between amino acids.)
(Streptomycin causes a misreading of mRNA.)
e. by inhibiting nucleic acid (______ and/or ____) synthesis
(Antiviral: AZT inhibits reverse transcriptase.)
(Antibacterial: rifampin inhibits RNA polymerase.)
(Antifungal: griseofulvin inhibits RNA polymerase.)
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
competitive
a. of an essential metabolite by _________ inhibition
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p.
77) wall
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid
cross- membrane
linkages of peptidoglycan) (p. 78)
c. by damagingprotein
the cellsynthesis
___________ (Polymixin cleaves the
layers of the membrane like a knife) (p. 78)
d. by inhibiting ________________ at 70s ribosomes (p. 79)
(Erythromycin inhibits translocase, freezing the ribosome on
the mRNA.)
(Tetracycline blocks tRNA attachment to mRNA)
(Chloramphenicol inhibits transferase, preventing peptide
bond formation between
DNA amino acids.)
RNA
(Streptomycin causes a misreading of mRNA.)
e. by inhibiting nucleic acid (______ and/or ____) synthesis
(Antiviral: AZT inhibits reverse transcriptase.)
(Antibacterial: rifampin inhibits RNA polymerase.)
(Antifungal: griseofulvin inhibits RNA polymerase.)
C. Criteria that determine the effectiveness of antimicrobial agents
1. Selective
________ toxicity = destroys or inhibits microbe without
affecting host cells
Spectrum
2. __________ of activity = range of microbes inhibited or killed
a. ______spectrum
Broad usually effective against Gram+ and Gram-
bacteria
(1). useful when no time to figure out exactly which microbe is
causing disease
(2). disadvantage is that it disrupts normal flora too (resulting
in secondary
Narrow__________ infections caused by opportunists).
b. _______spectrum requires identification of the pathogen
3. Tissue distribution, metabolism & excretion
Soluble
a. ______
Stable in body fluids (to be distributed in the blood)
b. _______ in body fluids (so it is not broken down easily)
assuring constantoral
and effective levels in the body (pH of
stomach may limit ______ administration unless coated)
absorbed
Half-life
c. must be _________ by body tissues affected
d. _________ refers tofrequency
the elimination rate of a drug
(this dictates the ___________ of dosage needed)
 allergenic and not cause adverse reactions
4. should be non __________
mutagenic to reduce development of resistant
5. should be non __________
strains
D. Disadvantages of antimicrobial therapy
toxic effects on normal tissues (especially liver &/or kidneys)
1. ______
normal flora
2. disturb ____________
Allergic reactions
3. ________
resistant strains of bacteria, usually by
4. development of __________
enzymes that destroy AMA (such as penicillinase)
producing _________
a.mutations
_________ occur naturally
b. resistance genes onplasmids
_________ that can be spread from
Conjugation
bacterial cells to other bacterial cells by ____________,
Transduction
______________, or Transformation
____________.
E. Avoid disadvantages by
Discriminate (careful) use of AMA
1. __________
identification of disease
a. Dr: proper ____________
prescription of AMA
microbe & proper __________
b. patient: maintain proper levels by
(1). taking medication at prescribed
intervals
_________
(2). taking medication for prescribed
time
length of _____
Synergistic effect of combination of 2-more AMA when
2. _________
resistance is likely to develop
disk-plate diffusion
F. AMA testing = _________________
method (p. 66)
1. procedure
lawn of
a. Inoculate a solid ______
bacteria on agar
b. Place paper disks saturated with
antibiotics on the surface
various _________
Incubate 24 hours and then observe
c. ________
2. The principle behind this is that during incubation, the antibiotic
inhibits growth of
diffuses into the agar and, if effective, ________
the bacteria in its presence.
3. observations
a. _________________
Zone of inhibition (no growth around the disk means the
AMA is effective)
Satellite colonies are isolated colonies in the zone of
b. _________
inhibition
resistant cells from the original population!
They represent ________
Overlapping antibiotics
(with synergistic
effects) may be needed
if satellite colonies
appear.

Location of
satellite colonies
if present