Chapter 9

Disinfectants , Antiseptics &

Sterilization
•Bacteriostat. An agent which prevents the reproduction and
multiplication of bacteria.
•Bactericidal : Kills bacteria .

•Biocides: include disinfectants, antiseptics & preservatives which are

chemical agents that have the ability to destroy or inhibit the growth of
m.o.

•Uses: medical, healthcare, livestock, environment, coatings,

plastics, pharmaceuticals, food & beverage manufacturing,
cosmetics.

•Disinfection: Is the process of removing microorganisms,

including potentially pathogenic ones, from the inert surfaces .

• Disinfectant not necessarily killing all microorganisms but

reducing them to a level acceptable for a defined purpose, e.g a
level which is not harmful to health nor to the quality of goods.
Disinfectant: should have bactericidal action to destroy
the bacterial population in order to make the environment free
from the ability to cause infection
• Chemical disinfectant have different level of disinfection

– High level disinfection: destruction of all m.o but not

necessarily bacterial spores. Some have good sporicidal
Activity.

– Intermediate level disinfection: destruction of all vegetative

bacteria including Mycobacteria tuberculosis but excluding
some viruses & fungi. They have little or no sporicidal
activity

– low level disinfection can destroy most vegetative bacteria,

some fungi and viruses.
• Antisepsis:

Is the destruction or inhibition of microorganisms on living tissues

having the effect of limiting or preventing the harmful results of
infection.
• They can be bacteriostatic agent.

• The chemicals used have adequate antimicrobial activity & are

applied to skin and mucous membranes, therefore they must not be
toxic or irritant for skin.
• Antiseptics are mostly used to reduce the microbial population on
the skin prior to surgery or on the hands to help prevent spread of
infection by this route.
• Antiseptics usually have lower concentrations than agents used
for disinfection
❖Factors affecting choice of antimicrobial
agent:
1. Properties of the chemical agent
2. Microbiological challenge
The type of m.o & the bioburden (microbial contamination). If the
bioburden is high, long exposure time or higher conc are needed.
M.o vary in their sensitivity to the action of antimicrobials .
3. Intended application whether the agent will be used for
preservation, antisepsis or disinfection.
4. Environmental factors Blood, body fluids, pus, milk, food
residues, even in small amounts, reduce the effectiveness of antimicrobial
agents to varying degrees and some are seriously affected. That’s why it is
important to clean instruments before disinfection.
5. Toxicity of the agent
✓ Vegetative bacteria
At in-use conc, disinfectants should be capable of killing most
vegetative bacteria (broad spectrum) within a reasonable contact
period including problematic m.o . G-ve bacilli are more
resistant than G +ve & responsible for most nosocomial
infections, where the most resistant is Pseudomonas
aeruginosa.

✓ Mycobacterium tuberculosis
• And other mycobacteria are resistant to many bactericides.
Resistance is either (a) intrinsic, mainly due to reduced cellular
permeability or (b) acquired
The greatest risk comes from undiagnosed patient.

Equipment used for respiratory investigation of a patient or carrier will

be contaminated & must be disinfected.
 ✓ Bacterial spores
• Bacterial spores are the most resistant of all microbial forms to
chemical treatment. (Prions?? Not m.o)
• Spore forming genera Bacillus & Clostridium
• The majority of antimicrobial agents have no useful sporicidal action.

✓ Fungi
• The vegetative fungal form is often as sensitive as vegetative bacteria to
antimicrobial agents. Fungal spores are more resistant but less resistant
than bacterial spores.

✓ Viruses
✓ Susceptibility of viruses to antimicrobial agents depends on whether the
viruses possess a lipid envelope. Non-enveloped viruses are frequently
more resistant to disinfectant , Hepatitis A virus is considered one of the
most resistant to disinfectants.
• Much concern for the hazards of HIV & HBV. Disinfectants must be able to
treat rapidly and reliably accidental spills of blood, body fluids or secretions
from HIV infected patients.

• It is recommended to use high level disinfectants for decontamination of HIV

or HBV infected reusable equipment.

✓ Prions
• Small (proteinaceous infectious particles) associated with causing
spongiform encephalopathies such as bovine spongiform
encephalopathy (BSE) in cattle and Creutzfeldt Jakob disease (CJD) in
humans.
• They are not m.o as they don’t have plasma membrane or nucleic
acids
• Risk of infectivity is highest in brain, spinal cord & eye tissues.

• Prions are resistant to most disinfectant procedures. Immersion in

sodium hydroxide .
❑ Alcohols

➢ As Disinfectants & antiseptics

✓ Aliphatic alcohols (Ethanol & isopropanol) are bactericidal against

vegetative bacteria+ mycobacteria. But not sporicidal.

✓ Activity drops sharply at Conc below 50%

✓ Properties: poor penetration of organic material (used in clean

conditions), reduce skin flora (prepare skin for injection or
procedure). Presence of water is eessential for activity so 100%
conc is relatively not effective.
❑ Aldehydes

• Highly effective biocides, they have sporicidal activity

Gluteraldehyde (highly active).

➢ Properties: broad spectrum of activity. Rapid rate of kill to

vegetative bacteria (few min), while spores ~few hrs, not
affected by organic matter.
➢ Uses: for chemical sterilization of medical and surgical
materials that cannot be sterilized by other methods.
➢ long time if HIV or Mycobacteria is suspected
➢ Formaldehyde Used in either liquid or gaseous state for
disinfection.
➢ Limitation of its use: It is irritant & not used for skin,
vapours are highly toxic (carcinogenic)
❑ Halogens (Chlorine & Iodine)
➢ Hypochlorite (bleach)
• Activity: They exhibit a rapid kill against a wide spectrum of
microorganisms, fungi and viruses. High conc of chlorine is
effective in eradicating acid-fast bacilli and bacterial spores.
• Disadvantage: they are corrosive, suffer inactivation by
organic matter and can become unstable.
• Advantage: are relatively inexpensive to use.

➢ Iodine I2
• Has a wide spectrum of antimicrobial activity. Gram-ve and
Gram+ve, bacterial spores (on extended exposure), mycobacteria,
fungi and viruses are all susceptible.

• Advantage: The antimicrobial activity of iodine is less dependent

than chlorine on temperature and pH

• Disadvantages: when used as antiseptic on skin causes staining of

skin
❑ Hydrogen peroxide & peroxygen compounds

• Are high level disinfectants

• H2O2 (oxidant & antiseptic) { H-O-O-H}

• Uses:

• antiseptic for open wounds & ulcers

• Disinfection of soft contact lenses as it is active against
Acanthamoeba which causes keratitis in contact lens wearers.
• 3-6% conc as general disinfectant
• At high concentrations (up to 35%) and increased temperature
hydrogen peroxide is sporicidal.
➢ Peracetic acid (CH3COOOH)

➢ Is the peroxide of acetic acid. A more potent biocide than

hydrogen peroxide, with excellent rapid biocidal activity
against bacteria, including mycobacteria, fungi, viruses and
spores.

• Combination of hydrogen peroxide & peracetic acid is

synergistic & is marketed as sterilant for dialysis machine

• Disadvantage: corrosive to some metals, highly irritant and

must be used in an enclosed system.
❑ Phenols

• Phenols are widely used as disinfectants.

• Advantages:
• As disinfectants they have good antimicrobial activity and
are rapidly bactericidal but generally are not sporicidal. They
are more active at acid pH.

• Disadvantages: The main disadvantages of phenols are

their caustic effect on skin and tissues and their systemic
toxicity.
➢ Bisphenols

Bisphenols are composed of two phenolic groups connected

by various linkages.

• Triclosan the most widely used. It is bacteriostatic at use-

concentrations and have little antipseudomonal activity. It has
application in medicated soaps, lotions & solutions, tooth
pastes & house hold products (fabrics & plastics).

• Concerns of bacterial resistance.

➢ Moist heat sterilization (autoclave)
• Usually involves the use of steam at temperatures in the range 121-
134°C.

• moist heat sterilization methods can only be achieved by the generation of

steam under pressure.

• Application: sterilization of culture media, dressings, sheets, surgical and

diagnostic equipment, containers .

• The most commonly employed standard temperature/time cycles are 121

°C for 15 minutes and 134°C for 3 minutes, respectively.

• Advantage of high temperature-short time cycles: lower fractional

degradation & achieving higher levels of sterility assurance.
• Steam as a sterilizing agent

• To act as sterilizing agent, steam should provide moisture and heat

efficiently to the article to be sterilized
➢ Dry heat sterilization
• Lethal effects of dry heat are due to oxidative processes which are less
effective than hydrolytic damage caused by steam sterilization.

• Thus, dry heat sterilization usually employs higher temperatures in the

range 160-180°C and requires exposure times of up to 2 hours depending
upon the temperature employed.

• Its application is generally restricted to glassware and metal

surgical instruments (where its good penetrability and non-
corrosive nature are of benefit)

• The instrument used is a hot air oven with perforated shelves to

allow for heat & air flow.
➢ Gaseous sterilization
• The chemically reactive gases ethylene oxide , and formaldehyde
possess broad-spectrum biocidal activity.

• Application: sterilization of re-usable surgical instruments, certain

medical, diagnostic and electrical equipment.

• Ethlyene oxide is more common internationally than formaldehyde.

• These techniques do not, offer the same degree of sterility assurance as

heat methods and are reserved for temperature-sensitive items.

• The sterilization processes are lengthy and therefore unsuitable for the
sterilization of high-turnover articles, even at the higher concentrations
and temperatures.

• They produce symptoms of acute toxicity including irritation of the skin

 ➢ Radiation Sterilization
• Includes accelerated electrons gamma-rays and
ultraviolet (UV) light
• Mechanism of action: The major target for these radiations is microbial
DNA, with damage occurring as a consequence of ionization (Gamma) or
excitation (UV light).

• UV light is less damaging and less lethal than ionization, therefore, UV

irradiation is not as efficient as electron or gamma-irradiation.

• Vegetative bacteria are the most sensitive to irradiation

• The resistance of the organism to radiation depends on the extent of
DNA damage required to produce cell death & the ability to carry out
effective repair
• In ionizing radiations (gamma-ray and accelerated electrons), microbial
resistance decreases with the presence of moisture or dissolved oxygen
(as a result of increased free radical production) and also with elevated
temperatures.
➢ Filtration sterilization
• The process of filtration is unique amongst sterilization
techniques in that
– it removes, rather than destroys, microorganisms.
– it is capable of preventing the passage of both viable and non-
viable particles and can thus be used for both the clarification and
sterilization of liquids and gases.
– Sieving can be regarded as absolute since it ensures the exclusion
of all particles above a defined size.
– Synthetic membrane filters, derived from cellulose esters or other
polymeric material
– Membrane filters of 0.2-0.22 µm nominal pore diameter are mainly
used.

Sterility test: a test which assesses whether a sterile pharmaceutical

or medical product is free from contaminating m.o by incubation of
either the whole or part of that product with nutrient medium. The
test is performed on a sample taken from a batch
❖ Disinfection policy

• The aim of the policy is to control the use of chemicals for disinfection and
antisepsis and to give guidelines on their use.

• The control of microorganisms is of prime importance in hospital and

industrial environments. Usually there is a committee who is responsible to
set & implement the policy (Infection control committee) . What
disinfectants to use, conc, where to use them (floors, ceiling, walls, air,
instruments), contact time.

• In hospitals categories of risk to patients may be assigned to equipment

that come in contact with him dictating the level of decontamination
needed.

• High risk items: have close contact with broken skin or mucous membrane
or are those introduced into a sterile area of the body. These should be
sterile. e.g sterile instruments, gloves, catheters, syringes and needles.

• Intermediate-risk items: are in close contact with skin or mucous

membranes and disinfection will normally be applied. e.g Endoscopes,
respiratory and anaesthetic equipment, wash bowls, bed-pans.

• Low-risk items or areas: walls, floors, etc., which are not in close contact
with the patient.