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Chapter 11: Physical and Chemical Control of Microbes
I. Joseph ______ started _________ techniques with medical 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 ControlA. _____________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.
B. __________means to reduce the number of pathogens (including viruses) until they are not a hazard, usually involving the use of antimicrobial chemicals.
C. _______________ refers to removing toxins. D. ____________ refers to a substantially
reduced microbial population that meets accepted health standards. A clean appearance is expected!
Lister aseptic pheno
l
Sterilization
Disinfection
Sanitization
Decontamination
E. Different situations warrant different levels of microbial control.1. daily life Simple ____________ with plain soap and water is considered to be the single most important step in preventing the spread of many infectious diseases!
handwashing
2. hospitals
Danger of ___________ (hospital acquired) infections because of:
a. _________ condition of hospitalized patientsb. higher concentration of sick people with ____________
microbes (*many resistant forms!!)c. _______ procedures (such as)
d. many health care workers are ______e. lack of _______ care (handwashing between patients, using gloves, etc.)
nosocomial
pathogenic
weakened
invasive
carriersasept
ic
3. microbiology/research/hospital laboratories must use ________ techniques a. Work surfaces should be ______. b. All media and instruments must be ______. c. Used ________ must be properly disposed of.
aseptic clea
n sterilecultures
III. Selection of an antimicrobial procedure depends on many factors such as the type of _______, the extent of ____________, ____________ conditions, and potential risk of _________.
A. types of resistant microbes 1. Bacillus and Clostridium can make ___________. 2. Mycobacterium has ______ cell walls. 3. ____________ is capable of metabolizing unusual
substances for food. (Like disinfectants!)
endosporeswax
y
microbe
contaminationenvironmen
talinfection
Pseudomonas
B. the extent of contamination (size of the microbial population) 1. ‘Industry standard’ requires that ____% of the population is
killed with every __ minutes of exposure to the treatmenta. 100 microbes 10 microbes 1 microbe in __ minutesb. 1010 microbes would take ___ minutes
SO, ________/_________ first helps reduce the population before disinfection or sterilization. C. environmental conditions 1. _____________ ( heat chemical action) 2. _____ 3. ____, _______, _______, ______ can all block chemical action
902
420
washing
scrubbing
temperaturepHdirt saliv
ablood
feces
D. Potential risk of infection1. _______ items come into direct contact with body tissues.Critic
al
2. ____________ items come into contact with mucous membranes, but do not penetrate body tissues.
Semicritical
3. ____________ items only touch keratinized skin surfaces.Noncritical
IV. Methods of Physical Control A. ______ works by_________ cell proteins /enzymes. It is the
most common control method because it is fast, reliable, inexpensive & nontoxic.
1. ______ heat a. _______ 100°C/10 minutes (kills most microbes &
inactivates most viruses, but does not destroy __________).
Heat
denaturing
Moist Boilin
g endospores
Pasteurization
b. ____________: a brief heat treatment followed by rapid cooling. (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
c. __________ (steam under pressure) (1). 15-20 psi/15-20 minutes/121°C (2). ________ equipment, media, etc. (3). used in canning procedures to destroy
Clostridium botulinum __________!
Autoclave
Sterilizes
endospores
2. ___ heat sterilizes.
a. Hot air ovens (160-170°C/2-3 hours) used when ________ is undesirable.
b. ____________ (burning)
(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 ______ during lab.
Dry
Incineration
flaming
Microbiology is Fun!
moisture
furnaces
incinerators
B. Radiation (waves having energy but no mass) causes lethal changes in DNA, denatures proteins, but doesn’t reliably destroy endospores)!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
2. Ionizing rays = ________ or _____________ 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 prevent ___________, to treat beeffor ________ contamination and used totreat chicken for _________ contamination.
ultraviolet
X-rays Gamma rayssteriliz
e
trichinosisE. coliSalmonella
3. microwaves a. do not affect microbes directly, but may kill by _____ they generate b. drawback is that microwave heating is ________
heat
uneven
C. Filtration (may be used for air, some heat sensitive materials such as serum, vaccines, drugs, IV fluidsbeer/wine) 1. _____ ________ ________ ____ (HEPA) filters remove
airborne contaminants; used in operating rooma, for people with
allergies, etc. 2. In fluid filtration, _______ are separated from ________ by passing through _______ 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)
High-Efficiency Particulate Air
solids
liquidsfilter
s
V. Methods of ________ Control (* for heat sensitive items, large surfaces)Destructive actions include injury to the cell _________,
denaturation of cell ________, inhibiting replication of _____.
A. Disinfectants Vs Antiseptics 1. _____________ are chemicals used on inanimate objects.
a. ___________ are chemicals that KILL/DESTROY germs. (examples: fungicides, bactericides, viricides)b. __________ refers to chemicals that do not kill, but prevent the growth of microbes .(examples: bacteriostatic, fungistatic)
2. __________ are disinfectants nontoxic enough to be used on skin. B. Germicides are grouped according to their _______ (strength) 1. __________ destroy everything, including endospores
(for sterilizing scalpels, respiratory therapy equipment, proctoscopes, plastic Petri dishes, endoscopes)
(ethylene oxide gas, hydrogen peroxide)
Chemical
membraneprotein
sDNA
DisinfectantsGermicid
esGermistatic
Antiseptics
potencySterilant
s
2. ____ level disinfectants (do not reliably destroy endospores) (used for GI endoscopes) (iodine, phenol, chlorhexidine, heavy metals such as silver nitrate) 3. ___________ level disinfectants (will kill Mycobacterium, but do not destroy all viruses or endospores, even with prolonged exposure) (used for stethoscopes, electrodes, thermometers) (alcohols: ethyl alcohol, isopropyl) 4. _____ level disinfectants (will not kill Mycobacterium)
(soaps, detergents)
High
Intermediate
Low
C. ______ _________ (5% Phenol is the standard against which chemical agents are tested and compared)
1. Each chemical is compared for the same length of _____ on the same _________ under ________ conditions 2. IF the chemical being tested requires a greater
____________ or a longer ______ than phenol, its efficiency is _____ than phenol.
IF the chemical being tested requires a lower concentration or a shorter time than phenol, its efficiency is _______ than
phenol.
3. Ratio of: tested chemical activityphenol activity
< 1 means _____ efficient than phenol > 1 means _____ efficient than phenol
Phenol coefficient
concentrationtim
e
greater
lessmore
timeorganis
midentical
less
D. Selecting the Appropriate germicidal chemical 1. ________ (the benefit of disinfecting or sterilizing an item or
surface must be weighed against the risks associated with the use
of that chemical) (hospital Vs home/office) 2. compatibility with the ________ being treated (metal, rubber,
glass, plastic) 3. ________ may necessitate rinsing 4. ______ and availability (bleach) 5. _________ and stability (concentrates require less space and
store for long periods, but when diluted/mixed, often have limited
shelf life) 6. _____________risk (disposal procedures needed)
Toxicity
material
ResidueCostStorage
Environmental
VI. Methods used for Preservation (delaying spoilage) of Perishable Products
A. ________ preservatives (both nonfood and food)1. organic ______ lower pH (inactivates enzymes, inhibits growth, but does not always destroy microbes)2. ________ and _______ inhibit germination of Clostridium
botulinum endospores! B. Low Temperatures
1. _____________ a. 0-10° C (___° C average) b. retards but does not prevent growth2. ________ a. ___° C b. prevents growth but does not kill all organisms
C. Increased _______ pressure by adding _____ or ______; causes water to leave the cell, killing it.D. ___________ (dehydration) of the material (natural [sun] or
artificial)
Chemical acid
snitrates
nitrites
refrigerator
freezer
osmotic
Desiccation
4
-20
salt
sugar
E. ____________ (freeze-drying)1. materials _______ frozen at temperatures well below 0°C2. vacuum while frozen to remove ________ (lightweight)3. biological cultures, medications, foods (expensive)
Lyophilization rapidl
y moisture
Chap 12: Elements of Chemotherapy
I. TerminologyA. ____________ = use of chemical agents to treat diseaseB. _________________ agent (CTA) = chemical agent used for treatment of disease (even cancer)C. _____________ agent (AMA) = chemical agent used to treat diseases caused by microbes
II. Antimicrobial AgentsA. Types of antimicrobial agents
1. ________ agents = metabolic products produced by certain groups of fungi and fungal-like bacteria
that are antibacterial in action 2. __________ agents = produced in the laboratory
3. _______________ agents = derivatives of natural agents altered in the laboratory by adding chemical groups
to improve effectiveness
ChemotherapyChemotherapeuticAntimicrobial
Natural
SyntheticSemi-synthetic
B. Modes of action 1. interfere with microbe’s chemosynthesis by inhibiting ________ 2. Disruption/interference with
a. of an essential metabolite by _________ inhibition (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.)
enzymes
competitive
B. Modes of action 1. interfere with microbe’s chemosynthesis by inhibiting ________ 2. Disruption/interference with
a. of an essential metabolite by _________ inhibition (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.)
enzymes
competitive
wall
B. Modes of action 1. interfere with microbe’s chemosynthesis by inhibiting ________ 2. Disruption/interference with
a. of an essential metabolite by _________ inhibition (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.)
enzymes
competitive
wall
membrane
B. Modes of action 1. interfere with microbe’s chemosynthesis by inhibiting ________ 2. Disruption/interference with
a. of an essential metabolite by _________ inhibition (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.)
enzymes
competitive
wall
membrane
protein synthesis
B. Modes of action 1. interfere with microbe’s chemosynthesis by inhibiting ________ 2. Disruption/interference with
a. of an essential metabolite by _________ inhibition (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.)
DNA RNA
enzymes
competitive
wall
membrane
protein synthesis
2. __________ of activity = range of microbes inhibited or killed a. ______spectrum 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
__________ infections caused by opportunists). b. _______spectrum requires identification of the pathogen
3. Tissue distribution, metabolism & excretion a. ______ in body fluids (to be distributed in the blood) b. _______ in body fluids (so it is not broken down easily)
assuring constant and effective levels in the body (pH of stomach may limit ______ administration unless coated)
c. must be _________ by body tissues affected d. _________ refers to the elimination rate of a drug
(this dictates the ___________ of dosage needed)
Spectrum Broa
d
secondaryNarro
wSoluble
absorbed
Stable
Half-life
oral
frequency
C. Criteria that determine the effectiveness of antimicrobial agents 1. ________ toxicity = destroys or inhibits microbe without
affecting host cells
Selective
4. should be non __________ and not cause adverse reactions5. should be non __________ to reduce development of resistant
strainsD. Disadvantages of antimicrobial therapy
1. ______ effects on normal tissues (especially liver &/or kidneys)2. disturb ____________3. ________ reactions4. development of __________ strains of bacteria, usually by
producing _________ that destroy AMA (such as penicillinase)a. _________ occur naturallyb. resistance genes on _________ that can be spread from
bacterial cells to other bacterial cells by ____________, ______________, or ____________.
allergenicmutagenic
toxic normal
floraAllergic resistan
tenzymesmutatio
ns plasmids Conjugati
onTransformation
Transduction
E. Avoid disadvantages by1. __________ (careful) use of AMA a. Dr: proper ____________ of disease
microbe & proper __________ of AMA b. patient: maintain proper levels by
(1). taking medication at prescribed _________
(2). taking medication for prescribed length of _____
2. _________ effect of combination of 2-more AMA when resistance is likely to develop
F. AMA testing = _________________ method (p. 66)
1. procedure a. Inoculate a solid ______ of
bacteria on agar b. Place paper disks saturated with
various _________ on the surface c. ________ 24 hours and then observe
Synergistic
disk-plate diffusion
lawn
antibioticsIncubat
e
Discriminate identificatio
n prescription
intervals
time
2. The principle behind this is that during incubation, the antibiotic diffuses into the agar and, if effective, ________ growth of the bacteria in its presence.
3. observationsa. _________________ (no growth around the disk means the
AMA is effective)b. _________ colonies are isolated colonies in the zone of
inhibitionThey represent ________ cells from the original population!
inhibits
Zone of inhibitionSatellite
Location of satellite colonies if present
resistant
Overlapping antibiotics (with synergistic effects) may be needed if satellite colonies appear.