Chapter 4

Physical Control of
Microbial Growth

Dr. Caridad D. Garcia

 Control of Microbial Growth
CONTROL =  Prevent
Kill disease
= Death &
reduction in number
infection transmission
&/or activity of the
total microbial flora Irreversible
 Prevent contamination
by or growth of
(permanent) loss of
remove
undesirable
inhibit the ability
microorganisms
to
kill reproduce
 Prevent [even
deterioration
Physical Methods &under optimum
spoilage of materials
Chemical Methods growth conditions]
 Recall
Lister started _________
Joseph ______ aseptic
techniques with surgical/medical
applications.
phenol -soaked
By using carbolic acid (_______)
rags and instruments during and after
surgery, gangrene and other infections
following surgery greatly diminished.
Definition of Terms
S_e_ _ _ _z_ _ _ _n
Gk: steria = barren
= inability to produce offspring

Killing or removing all forms of microbial life

(including endospores) in a material or an
object.
 Sterile (devoid of microbial life)
 Absolute (100%)
Sterilant: sterilizing agent

Commercial sterilization: sufficient heat

treatment to kill endospores of C. botulinum
in canned foods.
D_s_nf_ _ _ _ _n
Disinfectant
Chemical
Inanimate objects

Destruction/removal of vegetative or
non-endospore forming pathogens
Reducing the number of pathogenic
microorganisms to the point where they
no longer cause diseases
physical methods
chemical methods
 Antiseptic
Chemical
S_ _ sis: Animate objects
Gk “decay or putrid”
Indicates bacterial contamination
Growth of microorganisms in the body
Presence of microbial toxins in the
blood & other tissues
A_ _ _ _ _ _:
Absence of significant contamination
Antiseptic is used on
living tissues
______________
Disinfectant is used on
objects
______________
Degerming or Degermation:
M_ch_n_c_l removal of most
microbes in a limited area
Example: Alcohol swab on skin
S_n_t_z _ _ _ _ _n
Cleansing technique
to reduce contamination to safe levels
sanitary
meet public health standards
to minimize chances of disease transmission
Restaurants, Dairies, Breweries
Food Industry
_ _ _ _ _ _ _ technique
eliminate/exclude microorganisms
to prevent contamination

Medical Asepsis
Surgical Asepsis

to prevent bacterial contamination in

food industry
Control of Microbial Growth:

Definitions
Bacteriostatic Agent: An agent that _
_ _ _ _ _ _ _ the _ _ _ _ _ _ of
bacteria, but does not necessarily kill
them.
stasis: “to stop or steady”
Germicide or Microbiocide: An agent
that kills pathogens
Control of Microbial Growth:

Definitions
Bactericide: An agent that _ kills
____
bacteria. Most do not kill endospores.
Viricide: An agent that inactivates
_ viruses
_ _ _ _ _ _.

_Fungicide
_ _ _ _ _ _ _ : An agent that kills fungi.

Sporocide
_ _ _ _ _ _ _ _ _: An agent that kills
bacterial endospores or fungal spores.
Different situations warrant different
levels of microbial control.
A. DAILY LIFE

handwashing with plain

Simple (1.)____________
soap and water is considered to be the
single most important step in
preventing the spread of many
infectious diseases!
 B. HOSPITALS

nosocomial (hospital acquired) infections

Danger of (2.) ___________
because of:

weakened
(3.) _________ condition of hospitalized patients
Higher concentration of sick people with
pathogenic
(4.) ____________ microbes (*many resistant forms!!)

(5.) _______ procedures (such as)

invasive

(6.) Many health care workers are ______

(7.) Lack of _______ care (handwashing carriers
between patients,aseptic
using gloves, etc.)

Choices: Aseptic Carriers Invasive

Nosocomial Pathogenic Weakened
C. MICROBIOLOGY/RESEARCH/
HOSPITAL LABORATORIES

aseptic techniques.
must use (8.) ________

clean .
(9.) Work surfaces should be ______

(10.) All media and instruments must be

sterile .
_______
cultures
(11.) Used ________ must be properly disposed
of.

Choices:
Cultures Clean Sterile Aseptic
Effectiveness of Antimicrobial Treatment/
Factors that Affect Death Rate

 Length of exposure
 Number of microorganism
 Nature of the microorganism
 Environment [temperature, pH]
 Concentration of the agent
 Mode of Action of the agent
 Presence of solvents, interfering organic
matter & inhibitors
Rate of Microbial Death
Bacterial populations subjected to heat or
antimicrobial chemicals die at a constant
rate.

Death curve,
plotted
logarithmically,
shows this
constant death
Rate: 90% / min
rate as a straight
line.
Figure 7.1a
Length of exposure.
The number of viable
organisms remaining
in the population
decreases
logarithmically, giving
a straight-line
relationship on the
graph.
Effect of microbial load:
A higher load of
contaminants requires
more time to destroy
Time it takes to kill a
microbial population is
proportional to number
of microbes.

Only a fraction of
organisms die during
a given time interval
Nature of the organism
Relative resistance of
spores versus
vegetative forms

Microbial species and life

cycle phases (e.g.:
endospores) have
different susceptibilities
to physical and chemical
controls.
Action of the agent,
whether microbicidal
or microbistatic

How does it kill or

inhibit the
microorganisms?
 Organic matter may interfere with heat
treatments and chemical control agents.
– Saliva/sputum
– Blood
– Feces
– Vomitus

 Longer exposure to heat produces the same

effect as shorter time at higher temperature.
Selection of an antimicrobial procedure depends on many factors
such as the

microbe the extent of (2) ____________,

type of (1)________, contamination (3)

environmental infection
____________ conditions, and potential risk of (4)_________.

A. types of resistant microbes

endospores
(5) Bacillus and Clostridium can make ___________.

waxy cell walls.

(6) Mycobacterium has ______
Pseudomonas is capable of metabolizing unusual
(7) ____________
substances for food. (Like disinfectants!)

Choices: waxy microbe infection environmental

endospores contamination Pseudomonas
 B. the extent of contamination (size of the microbial
population)
90
‘Industry standard’ requires that (8) ____% of the
population is killed with every (9) __
2 minutes of exposure to
the treatment
100 microbes  10 microbes  1 microbe in (10) __ 4
minutes
1010 microbes      would take (11) ___ 20 minutes
washing or (13)_________
SO, (12) ________ scrubbing first helps
reduce the population before disinfection or sterilization.

C. environmental conditions
temperature
(14) _____________ ( heat  chemical action)
pH
dirt
(15) _____ saliva blood feces
(16) ____, (17) _______, (18) _______, (19)______
can all block chemical action
Choices: 90 4 2 20 pH dirt saliva
feces scrubbing blood washing temperature
D. Potential risk of infection
Critical
(20) _______ items come into direct contact with body tissues: (21-23)

Semicritical
(24) ____________ items come into contact with mucous
membranes, but do not penetrate body tissues. (24-26)
Noncritical
(27) ____________ items only touch keratinized skin surfaces. (28-29)

A B

E
D
C

F G H Semicritical
Critical
Noncritical

1. Critical; B; F; G 2. Semicritical; A; D; E 3. Noncritical; C; H

Cellular targets of control
1. Cell wall
2. Cell membrane
3. Cellular synthetic
processes (DNA, RNA)
4. Proteins

27
Surface Acting Agents lower
are wetting agents that lower Cell wall target-- -
 SURFACTANTS
the surface tension of a liquid, Surfactants agents
Surface-acting can
allowing easier spreading, that forms
insert a water-
in the
and lower the interfacial insoluble
tension between two liquids. lipoidal interface
layers to
•Soaps
disrupt it & create
 Ex: detergents,
abnormal
•Cetylpyridinium chloride wetting agents,
channels that alter
•Benzalkonium chloride dispersing agents,
permeability and
surface-tension
•Fatty alcohols cause leakage
depressant
both into and out
of the cell.
Mode of Action
Affecting Protein a. The native
Function (functional) state
is maintained by
bonds that create
active sites to fit
the substrate.
Some agents
denature the
protein by
breaking all or
some secondary
& tertiary bonds.
Mode of Action Affecting Protein Function

b. Complete
unfolding
c. Random bonding &
incorrect folding
d. Some agents react
with functional
groups on the
active site &
interfere with
bonding.
Practical concerns
 Does the application require sterilization?
 Is the item to be reused?
 Can the item withstand heat, pressure,
radiation, or chemicals?
 Is the method suitable?
 Will the agent penetrate to the necessary
extent?
 Is the method cost- and labor-efficient & is it
safe?
 Physical Methods of Control

Heat Radiation
 Moist [sterilization:  Ionizing [x-ray;
steam under pressure; cathode; gamma] –
disinfection [boiling sterilization
water; hot water,  Nonionizing [UV] -
pasteurization] disinfection
 Dry [Incineration;
dry oven]
Physical Control: HEAT
 Mode of action: denaturing their
enzymes and other proteins.
DENATURATION – loss of
normal characteristics due to molecular
alteration.
Heat resistance varies widely among
microbes.
Physical Control: HEAT
MOIST HEAT DRY HEAT
 Hot water, boiling  Air w/ low moisture
water, steam content has been
 lower temperature heated by a flame or
600 TO 1350C electric heating coil
 1600C 
 shorter exposure time
 Cell dehydration 
 Coagulation &
denaturation of protein oxidation ashes
Thermal Death Point (TDP): Lowest
temperature at which all of the microbes in
a liquid suspension will be killed in ten
minutes.
Thermal Death Time (TDT): Minimal
length of time in which all bacteria will be
killed at a given temperature.
Decimal Reduction Time (DRT): Time
in minutes at which 90% of bacteria at a
given temperature will be killed. Used in
canning industry.
Time Deaths/ Number of Decimal
(min) min Survivors Reduction
0 0 1,000,000 Time (DRT)
1 900,000 100,000
2 90,000 10,000 Minutes to kill
3 9,000 1,000 90% of a
4` 900 100 population at a
5 90 10 given
temperature
6 9 1
 Use of hot water or steam
MOIST HEAT

 Mode of action – Methods:

denaturation of 1. Steam under Pressure
proteins, 2. Live, non-
destruction of pressurized steam
membranes & 3. Boiling
DNA 4. Pasteurization
 sterilization
MOIST HEAT: 1a. Sterilization with Steam Under Pressure:
autoclave (Normal)15 psi/121oC/10-40min
[all vegetative cells & endospores are killed in about 15
minutes]
Autoclave: Closed Chamber with High Temperature and Pressure
 MOIST HEAT: 1. Sterilization with Steam Under
Pressure: autoclave 15 psi/121oC/10-40min

 Heat-resistant materials: glass wares, cloth

(surgical dressings), rubber (gloves),
metallic instruments, liquids, paper, some
media & some heat-resistant plastics
 Disposable plastic petri dishes
 Ineffective for sterilizing substances that
repel moisture: oils, waxes, powders
 MOIST HEAT: 2. Intermittent Sterilization or
TYNDALLIZATION

 Fractional (discontinuous) sterilization

 1000C; 30-60 min x 3 days
 Requires no pressure cooker
 Disinfection
 Heat-sensitive culture media e.g.,
containing sera, egg, carbohydrates
Jars fitted with a filter disc or a polyfill lid filter are
boiled or steamed at 212°F (100°C) for 30 min in
a pot with lid, three days in a row.

Between the boiling steps the jars are kept warm,

around 30°C(but room temperature will work too),
to allow the remaining endospores to germinate.

The basic principle behind this method is that any

resistant endospores will germinate after the first
heating and therefore be susceptible to killing
during the second and third heating.
Timetable of the tyndallization (=fractional sterilization)
process

1) Steam heating to 100 °C for 30 min

Vegetative cells are destroyed but endospores survive

2) Incubate at 30°C-37°C overnight

Most bacterial endospores germinate

3) Second heat treatment, 100 °C, 30 min

Germinated endospores are killed.

4) Second incubation at 30°C-37 °C overnight

Remaining endospores germinate

5) Third heat treatment, 100 °C, 60 min

Last remaining germinated endospores are killed
MOIST HEAT: 3. PASTEURIZATION

 Louis Pasteur
 Heat is applied to liquids to kill potential
agents of infection & spoilage, while at the
same time retaining the liquid’s flavor &
food value
 Beer, wine, milk, juices. Ice cream, yogurt
 Prevent transmission of milk-borne
diseases from infected cows or milk
handlers
 Thermoduric organisms survive
MOIST HEAT: 3. PASTEURIZATION
Primary targets are Non-spore forming pathogens

 Salmonella (food infection)

 Campylobacter jenuni (acute intestinal
infection)
 Listeria monocytogenes (listeriosis)
 Brucella species (undulant fever)
 Coxiella burnetii (Q fever)
 Mycobacterium bovis
 Mycobacterium tuberculosis
 Several Enteric viruses
 MOIST HEAT: 3. PASTEURIZATION

 Classic Method of Pasteurization

65oC x 30 minutes
630-660C x 30 min (batch method)
 MOIST HEAT: 3. PASTEURIZATION

 High Temperature Short Time

Pasteurization (HTST)
72oC x 15 sec (flash method)
Coxiella & Mycobacterium
Milk keeps well under refrigeration
Thermoduric organisms survive
 MOIST HEAT: 3. PASTEURIZATION

 Ultra High Temperature Pasteurization

(UHT)
140oC x 1-2 sec sterilized milk
Advantage: Milk can be stored at room
temperature for several months
(unopened).
Phosphatase test
Test to determine whether products have
been pasteurized

Phosphatase is an enzyme that is present

in milk & is destroyed by adequate
pasteurization.
MOIST HEAT: 4. BOILING
 Boiling water bath or chamber
 1000C x 30 min: tubercle bacilli;
staphylococci
 Hepatitis virus: survive up to 30 min
of boiling
 Endospores: survive up to 20 hr 
DRY HEAT: 1. DIRECT FLAMING

 Used to sterilize
inoculating loops
and needles.
Bunsen
Burner: 1,87000C
Heat metal until it
has a red glow.
DRY HEAT: 2. INCINERATION
 Destruction of microbes by
subjecting to extremes of
dry heat
Microbes ashes &
gas
 Incinerator: 8000-6,5000C
 Effective way to sterilize
disposable items (paper
cups, dressings) and
biological waste.
DRY HEAT:
3. HOT AIR STERILIZATION
 Oven
 1500-1800C [1700C] x 2-4 hours
 Glassware, metallic instruments,
powders & oils
 Not suitable for plastics, cotton &
paper w/c may burn or for
solutions w/c will dry out

Hot-air Autoclave
Equivalent
170˚C, 2 hr 121˚C, 15 min
treatments
 Cold/Low Temperature

Effect depends on the  Cold merely

microbe & method retards the
 Refrigeration activities of most
 Freezing microbes
 Lyophilization  -700 to -1350C
 REFRIGERATION
 Temperatures from 00 Pathogens that can
to 7oC. survive for several
 Bacteriostatic effect months in the
refrigerator:
 Reduces metabolic  Staphylococcus aureus
rate of most microbes  Clostridium species
so they cannot  Streptococcus species
reproduce or produce  Several types of yeasts,
toxins molds, & viruses
 Food, drug & culture
preservation
 FREEZING
Freezing: Temperatures below 0oC.
 Deep/Flash/Quick Freezing:

-500 to -950C
Does not kill most microbes
Food, drug & culture preservation
 Slow Freezing: More harmful because ice
crystals disrupt cell structure
 Over a third of vegetative bacteria may
survive 1 year.
 Most parasites are killed by a few days of
freezing.
 LYOPHILIZATION or FREEZE-DRYING
 Quick freezing (-540 to -720C) & water is
removed by a high vacuum
(sublimation). The container is then
sealed with a high-temperature torch
powderlike residue
 Mechanism of action: decreased
chemical reactions & changes in proteins
 Long term preservation of microbial
cultures; food & drug
 DESSICATION

 Removal of water
 Bacteriostatic
 Food preservation
 In the absence of water, microbes cannot grow or
reproduce, but some may remain viable for years.
After water becomes available, they start growing
again.
 Selica gel: dessicant
 DESSICATION

Susceptibility to dessication varies widely:

Neisseria gonnorrhea: Only survives
about one hour
Mycobacterium tuberculosis: several
months
Viruses are fairly resistant
Clostridium spp. and Bacillus spp.:
decades
As a general rule
chilling, freezing, and dessication
should NOT be construed as methods
of disinfection or sterilization because
their antimicrobial effects are erratic
& uncertain, and one cannot be sure
that pathogens subjected to them
have been killed.
 Osmotic Pressure
high concentrations of salts and sugars in foods
increase the osmotic pressure hypertonic
environment
 Plasmolysis: As water leaves the cell, plasma
membrane shrinks away from cell wall. Cell
may not die, but usually stops growing.
Yeasts and molds: More resistant to high
osmotic pressures
Staphylococci spp. that live on skin are
fairly resistant to high osmotic pressure
 RADIATION
 Energy emitted from atomic activities &
dispersed at high velocity through matter
or space.
 Ionizing Radiation – shorter wavelength;
more energy; deeper penetration
Gamma Rays
X Rays
Cathode Rays
 Nonionizing Radiation
Ultraviolet Rays
Forms of Radiation
A. Ionizing radiation can
penetrate a solid barrier,
bombard a cell, enter it &
dislodge electrons from
molecules. Breakage of DNA
creates massive mutations.
B. Nonionizing radiation enters
a cell, strikes molecules, &
excites them. The effect on
DNA is mutation by
formation of abnormal
bonds.
C. A solid barrier cannot be
penetrated by nonionizing
radiation.
 Ionizing Radiation:
1. GAMMA RAYS
 Wavelength: 10-3 nm
 Capable of deep penetration thus lethal
to all forms of life
 Mechanism: destruction of DNA
 Use: pharmaceuticals, medical & dental
supplies
 Require longer hours to sterilize large
masses
 Ionizing Radiation:
2. X RAYS or Roentgen Rays
 Lethal to all forms of life
 With considerable energy & penetration
ability
 Impractical to use in microbial control
- expensive
- difficult to use efficiently: radiations
are given off in all directions
 Ionizing Radiation:
3. CATHODE RAYS or
HIGH-ENERGY ELECTRON BEAMS

Bactericidal
Can sterilize packaged products:
pharmaceuticals, disposable dental
& medical supplies, plastic syringes,
surgical gloves, sutures, catheters,
etc.
Ionizing Radiation: ULTRAVIOLET RAYS

100 nm to 400 nm
240 nm to 280 nm: most lethal
[peak @ 260 nm]
Germicidal lamp: 254 nm
Ionizing Radiation: ULTRAVIOLET RAYS
Readily passes through air
slightly through liquids
poorly through solids
NOT as penetrating as ionizing
radiation
object to be irradiated must be
directly exposed for full effect
 Mechanism of Action
1. Formation of pyrimidine
dimers in adjacent:
cytosines
thymines
cytocine & thymine
inhibit correct replication of
DNA
mutation
inhibition of growth
DEATH

2. Formation of toxic ohotochemical products or

free-radicals: bind to DNA, RNA & Proteins
 Application of ULTRAVIOLET RAYS:
Disinfection

Germicidal lamp (99%): hospital

rooms, operating rooms, schools, food
preparation areas, dental offices
Disinfection of air: post-op area, food-
processing plants & slaughterhouses
Vaccines, plasma & other medical
products
Drinking water, milk, fruit juices
 Application of ULTRAVIOLET RAYS: Disinfection of water

Water flows though racks of UV lamps & is exposed to 254 nm UV radiation.

 Problems with UV Radiation

 dusts on lamps reduces effectiveness

Organic dirt reduces effectiveness
Intensity is dependent on proximity &
contact time
Personnel must be protected from over
exposure
Disadvantage of ULTRAVIOLET RAYS

Damaging effect of overexposure:

Retinal damage: kerato
conjunctivitis
Skin damage: Skin wrinkles;
sunburn; erythema; skin cancer
 Microwave Radiation

 Wavelength ranges from 1 millimeter to 1 meter

 Heat is absorbed by water molecules
 May kill vegetative cells in moist foods
 Bacterial endospores, which do not contain water,
are not damaged by microwave radiation
 Solid foods are unevenly penetrated by
microwaves
 Trichinosis outbreaks have been associated with
pork cooked in microwaves
 Sound waves or Ultrasonic Vibration
Usually used in water environment to
deliver sudden pressure changes near
the cells

It is delivered as an ultrasonic bath to

dislodge debris & clean instruments,
particularly those with multiple
channels that are difficult to clean
(bronchoscope, endospores scope)
Sound waves or Ultrasonic Vibration

Vegetative cells, particularly

gram negative organisms are
most sensitive

Many viruses & spore

forming bacteria are resistant
 Removal of microbes
FILTRATION by passage of a liquid
or gas through a
screen like material
(filter) with small
pores.
Mechanism: separation
of microorganisms
from liquid or gas
Use: to sterilize heat
sensitive materials
like vaccines,
enzymes, antibiotics,
and some culture
media.
 FILTRATION

 Membrane Filters; Molecular Filters;

Microbial Filters:
cellulose acetate or plastic polymer
ex: cellulose acetate, nitrocellulose
thickness: 0.1 nm
 Membrane Filtration
A. Vacuum Assembly for achieving
filtration of liquids through
suction.
Inset shows filter as seen in cross
section, with tiny passageways
(pores) too small for the
microbial cells to enter but large
enough for liquids to pass
through.
B. Scanning electron micrograph of
filter showing relative size of
pores & bacteria trapped on its
surface (5,900x).
Membrane Filtration of Liquids
 FILTRATION
Different pore sizes:
 Lice; yeast = 1 mm
 Parasites = 5 to 100 μm
 Bacteria = 0.22 and 0.45 μm
 Mycoplasma = 0.5 μm
 Viruses & some large proteins = 0.01 μm
 FILTRATION
High Efficiency Particulate Air Filters
(HEPA Filters)
 0.45 μm
 Use: air filtration
 OR; burn units
 High efficiency respiratory masks
 Biological cabinets
 VERY EXPENSIVE
HEPA Filtration of Air
Hands Spread Disease
 Seatwork: Identify the BEST PHYSICAL Method of
Control to be used.

1. Carcasses 8. Microorganisms
2. Drinking water in rural 9. Oils, waxes
areas 10. Metal equipment
3. Contaminated 11. Bronchoscopes
dressings & wipes 12. Operating room
4. Sterilized Milk 13. Vaccines
5. Linens, surgical drapes 14. Yogurt, beer, wine
6. Food 15. Fruits

7. Packaged gloves
Quiz: Methods of Physical Control
1. ______
Heat works by_________
denaturing cell proteins /enzymes.
It is the most common control method because it
is fast, reliable, inexpensive & nontoxic.
2. Moist heat uses water
______
Boiling 100°C/10 minutes (kills most microbes &
3. _______
inactivates most viruses, but does not destroy
endospores
4. __________).

Choices:
Denaturing Microorganisms Endospores Heat
Fungi Dry Moist
Cold Incineration Boiling Duplicating
 Pasteurizatio
5. ____________: a brief heat treatment followed
n
by rapid cooling. (Kills pathogens and reduces the
number of spoilage organisms in milk, juices, wine,
beer: Usually does not sterilize!)
6. UHT (Ultra High Temperature) ____0C in 30
minutes
7. HTST (___ ___ ____ ____) 72°C/15 seconds
8. Milk pasteurized by_______ method is
sterilized milk.
 Autoclaving
9. __________ (steam under pressure) 15-20 psi/15-20
minutes/121°C

Sterilizes equipment, media, etc.

10. ________
12. used in canning procedures to destroy endospores of the
Clostridium __________
bacteria _________ botulinum

Autoclave a closed-
12. ________:
chamber equipment that
delivers steam under
pressure.
 Dry
13. _____ heat sterilizes.
moisture
14. Hot air ovens (160-170°C/2-3 hours) used when ________
is undesirable.
Incineration (burning)
15. ____________
Incinerators or furnaces are used to destroy disposable
16 _________
items, soiled dressings, tissue specimens etc. @ 800°C to
6500°C
17. The hottest part of a Bunsen burner flame reaches 1,870°C
flaming during lab.
for ______

Microbiology
is Fun!
17. Radiation (waves having energy but no mass)
causes lethal changes in ( DNA or RNA),
denatures proteins, but doesn’t reliably
destroy endospores)!

ultraviolet radiation
18. Nonionizing rays = ________
can be used to reduce the number of
organisms in air and on clean surfaces but
of limited use, cannot penetrate materials
like cloth, glass, paper
X-rays or
19. Ionizing rays = ________
Gamma rays
20. ____________ can be used to
21. __________
sterilize items that are heat or
chemical sensitive, such as plastics more
effective, penetrates liquids and most solids
22. Radiation can be used to treat pork to
prevent
trichinosis
___________,
E. coli
23. to treat beef for ________ contamination
and
Salmonella
24. used to treat chicken for ___________
contamination.
25. Microwaves do not affect microbes directly, but may kill
by the (heat or cold) they generate.
26. The drawback of microwaves is that microwave heating is
(even or uneven).
27. Radiation or Filtration (may be used for air, some heat
sensitive materials such as serum, vaccines, drugs, IV
fluids,beer/wine)
28. _____________(HEPA) filters remove airborne
contaminants; used in operating room, for people with
allergies, etc.
 liquids are separated from
29. In fluid filtration, _______
solids
________ by passing through _______
filters with extremely
fine pores. Mechanical force or vacuum suction helps fluid
through the filter
small
30. It does not sterilize unless pore size is _______enough
to trap everything (smaller pores,  cost).

Lyophilization
31. ______________ or (freeze-drying)
rapidly or slowly
32. materials are ________________ frozen at
temperatures well below 0°C vacuum while frozen to remove
33. ________;
moisture used in (lightweight) biological cultures,
medications, foods and generally more expensive.
 Dessication (dehydration) of the material (natural [sun]
34. ___________
or artificial)

Osmotic
35. Increased __________ pressure by adding salt or sugar;
causes water to leave the cell, killing it.

Low Temperatures
Refrigeration
36. _____________ makes use of low temperature
4
37. from 0-10° C (___° C average) retards but does not prevent
growth
freezer
38. Its ________ compartment
-20
39. with _______° C temperature prevents growth but does
not kill all organisms.