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 means COMPLETE destruction of viruses and
A. __________
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 refers to removing toxins.
C. _____________
Sanitization refers to a substantially
D. __________
reduced microbial population that meets
accepted health standards.
A clean appearance is expected!
 Sanitized items (not sterilized!!)

E. Different situations warrant different levels of microbial control.

1. daily life
handwashing
Simple ___________with plain soap and water
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 condition of hospitalized patients
a. _________
pathogenic microbes
b. higher concentration of sick people with _________
(*and many resistant forms!!) surgery
invasive procedures (such as)
c. _______

carriers
d. many health care workers are ______
aseptic care (handwashing
e. lack of _______
between patients, using gloves, etc.)
3. microbiology/research/hospital laboratories
aseptic techniques
must use ________
a. Work surfaces should be ______
clean .
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 ___________.
waxy cell walls.
2. Mycobacterium has ______
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. _______
Surgical instruments Biopsy forceps

needles
Semicritical
2. ____________ items come into contact with mucous membranes,
but do not penetrate body tissues.
Respiratory equipment

Vaginal
speculum

Anal proctoscope
 Noncritical items only touch keratinized skin surfaces.
3. ____________

Sphygmomanometer
(blood pressure cuff)

stethoscope

IV. Methods of Physical Control

denaturing cell proteins /enzymes.
Heat works by_________
A. ______
It is the most common control method because it is
fast, reliable, inexpensive & nontoxic.
Moist heat
1. ______
Boiling 100°C/10 minutes (kills most microbes
a. _______
& inactivates most viruses, but does not destroy
endospores
__________).
 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 (UV)
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
trichinosis
pork to prevent ___________, to treat beef
E. coli contamination and used to
for ________
Salmonella contamination.
treat chicken for _________
 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
rooms, 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
proteins
denaturation of cell ________, DNA
inhibiting replication of _____.
A. Disinfectants Vs Antiseptics
Disinfectants
1. _____________ are chemicals used on inanimate
objects.
Germicides are chemicals that KILL/
a. ___________
DESTROY germs.
(examples: fungicides, bactericides, viricides)

Germistatic refers to chemicals that do

b. __________
not kill, but prevent the growth of microbes.
(examples: bacteriostatic, fungistatic)
Antiseptics are disinfectants nontoxic
2. __________
enough to be used on skin.
B. Germicides are grouped according to their _______
potency (strength)
Sterilants destroy everything, including endospores
1. __________
(for sterilizing scalpels, respiratory therapy equipment,
proctoscopes, plastic Petri dishes, endoscopes)
(ethylene oxide gas, hydrogen peroxide)
2. High
____ level disinfectants (do not reliably destroy
endospores) (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
4. ____
Mycobacterium) (soaps, detergents)
 Phenol coefficient
C. ______ _________ (5% Phenol is the standard against which
chemical agents are tested and compared)
time
1. Each chemical is compared for the same length of _____
identical conditions
organism under ________
on the same _________
concentration
2. IF the chemical being tested requires a greater ____________
less than
time than phenol, its efficiency is _____
or a longer ______
phenol.

IF the chemical being tested requires a lower concentration

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

3. Ratio of: tested chemical activity

phenol activity
less efficient than phenol
< 1 means _____
more efficient than phenol
> 1 means _____
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
2. compatibility with the ________
(metal, rubber, glass, plastic)
Residue may necessitate rinsing
3. ________
Cost and availability (bleach)
4. _____
Storage and stability (concentrates
5. ________
require less space and store for long
periods, but when diluted/mixed, often
have limited shelf life)
Environmental
6. _____________risk
(safe disposal procedures needed)
VI. Methods used for Preservation (delaying spoilage) of Perishable
Products
A. Chemical
________ preservatives (both nonfood and food)
acids lower pH (inactivates enzymes, inhibits
1. organic ______
growth, but does not always destroy microbes)
nitrates and _______
2. ________ nitrites inhibit germination of Clostridium
botulinum endospores!
B. Low Temperatures
refrigerator
1. _____________
4
a. 0-10° C (___° C average)
b. retards but does not
prevent growth
freezer
2. ________
a.-20
___° C
b. prevents growth but
does not kill all
organisms
C. Increased _______
osmotic pressure by adding _____
salt
sugar causes water to leave the cell, killing it.
or _____;

Desiccation (dehydration) of the material

D. ___________
(natural [sun] or artificial)

Lyophilization (freeze-drying)
E. ____________
rapidly frozen at temperatures well below 0°C
1. materials _______
moisture (lightweight)
2. vacuum while frozen to remove ________
3. biological cultures, medications, foods (expensive)
 Elements of Chemotherapy
I. Terminology
Chemotherapy = use of chemical agents to treat disease
A. ____________
Chemotherapeutic agent (CTA) = chemical agent used for
B. _______________
treatment of disease (even cancer)
Antimicrobial agent (AMA) = chemical agent used to treat
C. ___________
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 agents = produced in the laboratory
2. _________
Semi-synthetic agents = derivatives of natural agents
3. _____________
altered in the laboratory by adding chemical groups to
improve effectiveness
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
competitive inhibition
a. of an essential metabolite by _________
(Sulfa drugs mimic PABA, blocking folic acid synthesis) (p. 77)
Competitive Inhibition

Folic acid

PABA
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
wall
b. by weakening/disrupting the bacterial cell ______
(Penicillin inhibits the enzyme that builds the amino acid cross-
linkages of peptidoglycan) (p. 78)

Glycan
“backbone”
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
membrane (Polymixin cleaves the
c. by damaging the cell ___________
layers of the membrane like a knife) (p. 78)

hydrophilic
Amphipathic
hydrophobic
B. Modes of action
enzymes
1. interfere with microbe’s chemosynthesis by inhibiting ________
2. Disruption/interference with
protein synthesis at 70s ribosomes (p. 79)
d. by inhibiting ________________
(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.)
 DNA and/or ____)
e. by inhibiting nucleic acid (______ RNA 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 of activity = range of microbes inhibited or killed
2. ________
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 infections caused by opportunists).
_________
Narrow
b. _______spectrum requires identification of the pathogen
3. Tissue distribution, metabolism & excretion
Soluble in body fluids (to be distributed in the blood)
a. ______
Stable in body fluids (so it is not broken down easily)
b. _______
assuring constant and effective levels in the body (pH of
oral administration unless coated)
stomach may limit ____
absorbed by body tissues affected
c. must be _________
Half-life refers to the elimination rate of a drug
d. _________
frequency
(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 _________
mutations occur naturally
a. _________
plasmids that can be spread from
b. resistance genes on _________
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 method (p. 66)
F. AMA testing = _________________
1. procedure
lawn of bacteria on agar
a. Inoculate a solid ______
antibiotics on the
b. Place paper disks saturated with various _________
surface
Incubate 24 hours and then observe
c. ________
2. The principle behind this is that during incubation, the antibiotic
inhibits growth of the
diffuses into the agar and, if effective, ________
bacteria in its presence.
3. observations
Zone of inhibition
a. _________________ (no growth around the disk means the
AMA is effective)
Satellite colonies are isolated colonies in the zone of
b. _________
inhibition (They represent
resistant
________ cells from the original
population!)
Location of
satellite colonies
if present
Overlapping antibiotics (with
synergistic effects) may be needed
if satellite colonies appear.