Disinfection is the elimination of pathogens,

except spores, from inanimate objects

 Disinfectants are chemical solutions used
to clean inanimate objects
Disinfection

(physical processes, e.g., UV radiation,

may also be employed to effect
disinfection)
 Germicides are chemicals that can be
applied to both animate (living) and
inanimate objects for the purpose of
eliminating pathogens
 Antiseptics are formulated for application
to living tissue
 The Ideal Disinfectant
 Resistant to inactviation
 Broadly active (killing pathogens)
 Not poisonous (or otherwise harmful)
 Penetrating (to pathogens)
 Not damaging to non-living materials
 Stable
 Easy to work with
 Otherwise not unpleasant
 Disinfectant Performance…
 Is dependent on Disinfectant concentrations
 Is dependent on length (time) of administration
 Is dependent on temperature during administration
(usual chemical reaction 2x increase in rate with each
10°C increase in temperature)
 Microbe type (e.g., mycobacteria, spores, and certain
viruses can be very resistant to disinfection—in general
vegetative cells in log phase are easiest to kill)
 Substrate effects (e.g., high organic content interferes
with disinfection—stainless steel bench easier to
disinfect than turd)
 It is easier (and faster) to kill fewer microbes than many
microbes
 Cleansing is the removal of soil or
organic material from instruments
Cleansing

and equipment & may be done,

clinically, in four steps:
 Rinsing the object under cold water
 Applying detergent and scrubbing object
 Rinsing the object under warm water
 Drying the object prior to sterilization or
disinfection
 Sterilization is the total elimination of all
microorganisms including spores
 Typically the last things to die are the
highly heat- and chemical-resistant
Sterilization

bacterial endospores
 Instruments used for invasive procedures
must be sterilized prior to use
 Moist heat or steam, radiation, chemicals
(e.g., glutaraldehyde), and ethylene oxide
(a gas) are employed for sterilization
 Sterilization by autoclaving, which uses
moist heat, is used in most hospital and
microbiology laboratory settings
  Sanitization: Lowering of microbial counts
to prevent transmission in public setting
(e.g., restaurants & public rest rooms)
Other Terms

 Degerming: Mechanical removal of

microbes, e.g., from hands with washing
 Sepsis: Bacterial contamination
 Antisepsis: Reduction of or Inhibition of
microbes found on living tissue
 Germincides, Fungicides, Virucides
 Physical versus Chemical disinfectants
 Static (stasis) versus Cidal (e.g.,
bacteriostatic versus bacteriocidal)
Exponential Death
10
50°C
Log Cell # 9
60°C
8 80°60
C °C

0
10 20 30 40
Time.
Different Kinds of Bacteria “Death”
Total cell count

1. Bacteriostatic
Viable cell count

Log Cell #
2. Bacteriocidal

3. Bacteriolytic

Time
  Gram-negative bacteria (with their outer membrane) are
generally more resistant than gram-positive bacteria to
disinfectants and antiseptics
Resistance to Killing
 Stationary-phase (I.e., non-growing) bacteria generally are
more resistant than log-phase (I.e., growing) bacteria
 Mycobacteria, endospores, and protozoan cysts and
oocysts are very resistant to disinfectants and antiseptics
 Nonenveloped viruses are generally more resistant than
enveloped viruses to disinfectants and antiseptics
 Organic matter (such as vomit and feces) frequently
affects the actions of chemical control agent
 Disinfectant activity is inhibited by cold temperatures
 Longer application times are preferable to shorter
 Higher concentrations, though, are not always preferable
to lower concentration (e.g., alcohols)
 Agent Mechanisms of Action Comments
Surfactants Membrane Disruption; Soaps; detergents
increased penetration
Quats (cationic Denature proteins; Antiseptic - benzalconium
detergent) Disrupts lipids chloride, Cepacol; Disinfectant
Antimicrobials
Organic acids High/low pH Mold and Fungi inhibitors; e.g.,
and bases benzoate of soda
Chemical

Heavy Metals Denature protein Antiseptic & Disinfectant;

Silver Nitrate
Halogens Oxidizing agent Antiseptic - Iodine (Betadine)
Disrupts cell membrane Disinfectant - Chlorine
(Chlorox)
Alcohols Denatures proteins; Antiseptic & Disinfectant
Disrupts lipids Ethanol and isopropyl
Phenolics Disrupts cell membrane Disinfectant
Irritating odor
Aldehydes Denature proteins  Gluteraldehyde - disinfectant
(Cidex); Formaldehyde -
disinfectant
Ethylene Oxide Denaturing proteins Used in a closed chamber to
sterilize
Oxidizing agents Denature proteins Hydrogen peroxide – antiseptic;
Hydrogen peroxide –
Surfactants
  Soaps are sodium or potassium salts of
Soap & Detergents
fatty acids, a natural product
 Detergents, instead, are artificial
surfactants
 While soaps are always negatively
charged, some detergents are negatively
charged while others are positively
charged
 One example of a positively charged
detergent are quaternary ammonium
compounds (a.k.a., quats)
  Quats are cationic detergents that act by
disrupting lipid bilayers
 Quats are bactericidal, fungicidal, viricidal
(enveloped), and amoebicidal
 Quats are most effective against Gram-
Quats

positive bacteria
 Quats do not kill endospores, Mycobacteria
spp., nor non-enveloped viruses
 Quats are rapidly inactivated by organics
including cotton and soap
 Zephiran  Benzalkonium chloride
 Cepacol Cetylpyridinium chloride
  Ag, Cu, Hg, Ni, Zn, Ag(NO3)2, CuSO4,
ZnCl2, HgCl2
Heavy Metals

 These metals (and metal ions) react with

sulfhydral (–SH) groups of proteins,
denaturing proteins
 Silver nitrate is used to treat Ophthalmia
neonatorum in newborns as caused by
Neisseria gonorrhoeae
 Oligodynamic action: the ability of very
small amounts of heavy metals (especially
silver and copper) to exert antimicrobial
activity
  Halogens are the seventh (VII) column of the
periodic table of elements
 Two halogens are regularly employed as
antimicrobials: Iodine and Chloride
 Iodine: commonly used as an antiseptic
Halogens

against all microbes, fungi, and viruses

 Iodine: It inhibits protein synthesis and
oxidizes –SH groups of amino acids
 Chlorine: Used as a disinfectant (10% bleach)
 Chlorine: Hypochlorous acid (HOCl) is a
product, formed in water, that is the active
form of the disinfectant
 Chlorine: Applied in treatment of drinking
water, swimming pool, and sewage
 Chlorination
 1744 discovered in Sweden
 1810 identified as an element
 1835 first used to control odors
 1890’s started to be used as a disinfectant
 1896 earliest recorded use in experiments on water
supplies
 1897 used in England to sterilize water mains following
typhoid outbreak
 1902 first continuous use in water supplies in Belgium
 1909 liquid chlorine (compressed gas) became
commercially available
 Subsequent rapid spread in use of chlorine throughout the
world
 WWI: Chlorine gas used as chemical warfare agent
 Chlorination
“What
Hypochlorite
is known mayaseither
modernbe added directly
chemical (i.e., inbegan
warfare the
form of
during bleach)
World WarorI.created
The firstwithin wateragent
chemical by bubbling
to be used
chlorine gas through the water
was large amounts of chlorine gas, about one hundred
sixty
Chlorine
tons, gas - preferred
which for medium
was released from to largepressurized
6,000 disinfection
systems into the wind by the Germans against the
cylinders
Allies.
Sodium TheHypochlorite
chlorine floated in a
(liquid) huge clouds
- typically used toward
for smallthe
disinfection
Allies systemsthe
until it reached andAllied
largelines
swimming
causingpools
men to die
from
Calciumthe effects of the(powder,
Hypochlorite chlorinetablet)
gas. -Because of the
typically used
large amounts
for private of gaspools
swimming released the chlorine caused
large amounts of yellowish fluid to form in the lungs of
 For water purification, do not use scented bleach
its victim, also causing eye, nose, and throat burning
before
Bromine sometimes
causing death byused as a less-smelly alternative
choking.”
 http://www.geocities.com/CapeCanaveral/Lab/4239/chemweapons/history.html
Hypochlorite is less effective in the presence of
significant organic compounds
  Aqueous ethanol (60-95%) and isopropanol
are used as disinfectants
 Effectively kill bacteria and fungi but not
endospores nor nonenveloped viruses
 Fast acting, no residue (evaporate away),
Alcohols

no staining
 But not very penetrating and no residual
activity (once gone gone)
 Exert their action by denaturing proteins and
dissolving lipids
 In tinctures, they enhance the effectiveness
of other antimicrobial chemicals
 Flammable; also may damage rubber,
plastic, etc.
  “Formulae: Fresh juice of
Organic Habanero peppers,
New Mexico Jalapeno, African
Bird peppers and Hatch Chili
peppers.
 Dosage: Five to thirty drops,
three times daily.  CAUTION ~
Tincture

EXTREMELY HOT!!
 Therapeutic Action: Cayenne
is the greatest herbal aid to
circulation and should be used
on a regular basis.  The extract
is very concentrated and gets
into the bloodstream quickly and
makes it a perfect first aid
remedy for heart attacks, stroke,
fainting, shock, dizziness,
hemorrhage, internal and
external bleeding.  Use a few
drops to 10 droppers full.  It has
saved many lives.
  “Formulae: Fresh Garlic Juice, Goldenseal root, Usnea lichen,
Myrrh gum, Pine resin, Echinacea root juice, Tea Tree oil,
Kelp, Black Walnut inner hulls, Oak galls and Cayenne pepper
in 80% grain alcohol.
 Dosage: Generally for external use but can be used in the oral
Tincture II

cavity.  Soak a cotton swab in the tincture and scrub into the
infected area, let air dry.  It has a burning sensation. If the
wound is tender, just flush it with multiple droppers full of the
A tincture
tincture but no needistoascrub
nonvolatile
it in. substance
(medicine)
 presented
Therapeutic Action: asnever
There has an alcohol solution,
been an infected occur
when e.g., (for fun
this formula withused. 
has been numerous [sic])…
It's excellent for treating any
cut or wound and it is anti-bacterial, anti-viral and anti-fungal. 
The tree resins in this formula leave an invisible protective,
anti-bacterial coating over the wound.  This formula was use
on a man in England who had the top of his knee torn off in an
automobile accident.  In 24 hours it literally glued his knee
back together.  A nurse from Ireland on the scene said in all
the years in the hospital, she had never seen such a severe
wound close right up and heal, and with no infections.”
Iodine & Iodophores

Phenol, Carbolic Acid, & Phenolics
 Phenol (carbolic acid) and derivatives
 Affect plasma membrane, inactivates
enzymes, and denature proteins
 Stable, persistant, and especially effective
when dealing with disinfecting materials
contaminated with organics…
 … but leave residual films, can irritate
skin, don’t kill endospores, and are
corrosive to rubber and plastics
 Some phenolics are mild enough for use
as antiseptics while others are too harsh
or otherwise dangerous to be employed
on living tissue
 Hexachlorophene, Triclosan, Lysol, soap
  HOOH, hydrogen peroxide, is most common
 HOOH is not a terribly effective disinfectant or
anticeptic
Oxidizing Agents

 This is because bacteria and body tissues

contain enzymes (catalase) that inactivate
hydrogen peroxide
 However, the oxygen released upon
inactivation can help oxygenate deep wounds
and thus kill strict-anaerobe contaminants,
e.g., Clostridium tetani
 Ozone and peracetic acid are also oxidizing
antimicrobial agents
 They exert their effect by oxidizing cell
macromolecules (e.g., proteins, DNA, etc.)
 Glutaraldehyde is
capable of effectiving
Glutaraldehyde
sterilization—at room
temperature, even
against endospores,
and even in the
presence of organics,
but achieving
sterilization requries
many hours of
exposure… and it is
nasty stuff to work
with!
Gaseous Chemosterilizers
 Propylene oxide (C3H6O)
 Chlorine gas (Cl2)
 Chlorine dioxide (ClO2)
 Ozone (O3)
 Ethylene oxide (C2H4O)…
 …is used to sterilize heat- or moisture-sensitive
items
 …is used for items damaged by heat or moisture
 …is not corrosive, not damaging to delicate
instruments, microscopes, disposable plastic
instruments and materials
 …permeates porous materials
 …dissipates rapidly from material
 …but is costly, toxic, carcinogenic, explosive, and
 Agent Mechanisms of Action Comments
Moist Heat, boiling Denatures proteins Kills vegetative bacterial cells and
viruses Endospores survive
Moist Heat, Denatures proteins 121°C at 15 p.s.i. for 30 min kills
Autoclaving everything
Moist Heat, Denatures proteins Kills pathogens in food products
Antimicrobials
Pasteurization
Dry Heat, Flaming Incineration of Used for inoculating loop
contaminants
Physical

Dry Heat, Hot air Oxidation & Denatures 170°C for 2 hours; Used for
oven proteins glassware & instrument sterilization
Filtration Separation of bacteria Used for heat sensitive liquids
from liquid (HEPA: from
air)
Cold, Desiccation and low Used for food & drug preservation;
Lyophilization temperature Does not necessarily kill so used for
(also desiccation) Long-term storage of bacterial cultures
Cold, Refrigeration Decreased chemical Bacteriostatic
reaction rate
Osmotic Pressure, Plasmolysis of contaminants Used in food preservation (less
Addition of salt or effective against fungi)
sugar
Radiation, UV DNA damage (thymine Limited penetration
dimers)
Radiation, X-rays DNA damage Used for sterilizing medical supplies
Strong vis. Light Line-drying laundry
 Application of Heat
 Heat is frequently used to kill microorganisms
 Thermal death point (TDP) is the lowest temperature at which all
bacteria in a liquid culture will be killed in 10 minutes
 Thermal death time (TDT) is the length of time required to kill all
bacteria in a liquid culture at a given temperature
 Decimal reduction time (DRT) is the length of time in which 90%
of a bacterial population will be killed at a given temperature
(especially useful in canning industry)
 Dry heat kills by oxidation (slow, uneven penetration)
 Moist heat kills by protein coagulation (denaturation) so requires
lower temperatures or shorter times, but the moisture must
penetrate to pathogens to be effective (grease & oil can block)
 Moist Heat
 Moist heat kills microbes by denaturing enzymes
(coagulation of proteins)
 Boiling (at 100°C, I.e., at sea level) kills many vegetative
cells and viruses within 10 minutes
 Autoclaving: steam applied under pressure (121°C for 15
min) is the most effective method of moist heat sterilization
—the steam must directly contact the material to be
sterilized
 Pasteurization: destroys pathogens (Mycobacterium
tuberculosis, Salmonella typhi, etc.) without altering the
flavor of the food—does not sterilize (63°C for 30 seconds)
 Higher temperature short time pasteurization applies higher
heat for a much shorter time (72°C for 15 seconds)
 An ultra-high-temperature, very short duration treatment
(140°C for 3 sec.) is used to sterilize dairy products
Sterilization Times
 171o C, 60 minutes, dry heat
 160o C, 120 minutes, dry heat
 149o C, 150 minutes, dry heat
 141o C, 180 minutes, dry heat
 121o C, 12 hours, dry heat
 121o C, 15 minutes, moist heat (but
don’t start the clock until entire item is
up to temp—e.g., large volumes fluid)
Filtration: Air & Fluids
Evaluating Disinfectants  Phenol Coefficient…
 …compares efficacy to that of phenol, with
greater efficacy indicated with coefficient >1
 …Salmonella typhi and Staphylococcus
aureus commonly used to determine
coefficients
 Filter-Paper/Disk Diffusion method…
 …placement of disinfectant impregnated filter
paper on well-inocated agar
 Use-Dilution test…
 …drying bacteria to surface followed by
exposure to disinfectant and subsequent
washing and inoculation of sterile broth
Disk Diffusion Method

Hypochlorous acid

Phenol

Nisin
Escherichia coli
Lysol
Hands Spread Disease
Link to Next Presentation
Acknowledgement

http://www.sw.vccs.edu/rth/RTH_132/decont.ppt
http://www.nt.gov.au/powerwater/html/pdf/tennant_creek_chlorination.ppt
http://www.bugcruncher.com/ws/docs/lekcii/D&S_INQUAL_eng.ppt (nice overview of specific antimicrobials)
s

http://www.delmarhealthcare.com/olcs/white/ppts/chapter%2021.ppt (nice overview of microbilogy in nursing)

http://www.countyofkings.com/health/ehs/forms/FDHNDLR.ppt
http://bioeng.ttu.edu.tw/~chunyi/microbes/Chapter7.ppt
http://microbiology.okstate.edu/courses/micro2124/babus/Chpt7_files/chapter7.ppt
 Chemical Antimicrobials

* Type of Disinfectant: H = High level; I = Intermediate level; L = Low level