Microbiology SDL

Disinfection
Definition:
1. The killing of many, but not all, microorganism. Pathogens must be killed,
but some organisms & bacterial spores may survive.
2. Process of removing most, but not all, viable organisms.
Classification
1. Antiseptics:
To reduce number of viable organism on the skin
Can be safely applied on the skin or mucous membrane, used to
prevent infection by inhibiting growth of bacteria
2. Bactericidal agents/Germicides:
Disinfectants which are able to kill bacteria
3. Bacteriostatic agents
Disinfectants which prevent multiplication of bacteria
Types, groups to which they belong, mode of action and uses of disinfectants:
Types
1.
2.
3.
4.
5.
6.
7.

Mode of Actions
Alcohols
Detergents
Phenols
Halogen (Chlorine&Iodine)
Heavy Metals
Hydrogen Peroxide
Aldehydes (Formaldehyde &

Glutaraldehyde)
8. Ethylene Oxide
9. Acids & Alkalis
10.Dye

Disruption Of Cell Membranes

Modifications of Proteins

Modifications of Nucleic Acid

1. Alcohols
Bactericidal
Ethanol is widely used to clean the skin before venepuncture or

immunization
Disorganises lipid structure in membranes
Denature proteins

Ethyl alcohol (ethanol) and isopropyl alcohol:

a. Antiseptic

b. Disinfect clinical thermometers

Methyl alcohol:
a. Treating cabinets & incubators affected by fungal spores

2. Detergents or Surface-active agents

Bactericidal
Composed of a long-chain, lipid soluble, hydrophobic portion and a

polar hydrophilic group

Can be cation, anion, or non-ionic group
Interact with the lipid in the cell membrane and disrupt the

membrane
Reducing surface tension
Used as wetting agents, detergents and emulsifiers

Cationic compounds
Most important
Enter cell & denatures proteins
Act on phosphate group on membraneLoss of semi-permeabilityProtein
denature
Group
Quaternary ammonium

Examples
Benzalkonium chloride,

Note
Active in alkaline. Better

compounds

cetyltrimethylammoniu

action against Gram-

m bromide

than Gram+. As skin

disinfectant. Inactivated
by hard water& organic
materials. No action on
spores, Tubercle bacilli,
& most viruses

Anionic compound
Such as soap, moderate action
Made up of Saturated or Unsaturated Fatty Acid
More effective on:
o Gram- : Saturated FA
o Gram+: Unsaturated FA
Amphoteric (Tego compound)

 Active against wide range of Gram- & Gram+

Less frequently used

3. Phenols (Carbolic acid)

1st disinfectant used in operating room (Lord Lister, 1860s)
Bactericidal
Nowadays rarely used due to its corrosive, toxic, caustic,

carcinogenic, and neurotoxic effect

Wide bactericidal range
Cause cell membrane damagerelease cell contentscause cell

lysis
Precipitate proteins
Inactivate membrane bound oxidases & dehydrogenases enzymes
Pseudomonas grow readily in Chloroxylenol

solution(disadvantages)
Nowadays replaced by phenol substitutes

Cresol

More active & less toxic. Surface

Lysol

disinfectant.
Disinfectant in hospital & domestic
practice. Hand washes. 50% soapy

Chloroxylenol (Dettol)

emulsion of cresol
Non-corrosive, non-irritant. Can
penetrate epithelial surface.

Hexachlorophene

Pseudomonas grow readily in it

In germicidal soaps. Possible
neurotoxicity (cerebral oedema &

Chlorexidine

convulsions)
Hand disinfectant & cleansing wounds

4. Halogens
Bactericidal
Modifies proteins
Iodine

Most effective skin disinfectant. Bind to

-Tincture: Can be irritating

tyrosine residue in proteins.

-Iodophores: Less irritating

Chlorine

Disinfection of water supplies, swimming

pools & food & dairy. Organic chloramines
are used in wound dressing.

Cross-linking essential sulfhydryl groups in enzymes to form the

inactive disulphides

5. Heavy Metals or Metallic Salts

Bactericidal
Modification of proteins (enzymes)
Bind to sulfhydryl groups, thereby blocking enzymatic activity
Mercury (Hg) and Silver (Ag) have the greatest antibacterial activity
of heavy metals
Mercury salts

Bacteriostatic. Used as mild skin

-Thiomersal/Merthiolate, Phenyl

antiseptics

mercury nitrate &

merbomin/mercurochrome
Silver salts

Silver nitrate drops useful to prevent

gonococcal ophthalmia neonatorum.
Silver sulfadiazine is used to prevent
infection of burn wounds
As fungicides

Copper salts

6. Hydrogen Peroxide (HO)

Clean wounds & disinfect contact lenses
Limited by ability of organism to produce catalase breakdown of

HO
Attack sulfhydryl groups, thereby inhibiting enzymatic activity

7. Aldehydes
Bactericidal, sporicidal, lethal to virus in aqueous solutions
Denature proteins & nucleic acid
Formaldehyde (gas form)

Preserve anatomical specimens

Destroying anthrax spores in hair & wool

Formalin (solution form)

Sterilising clean metal instrument

Sterilising instruments & heat-sensitive catheters

Glutaraldehyde

Fumigating wards, sick rooms & laboratories

10x more effective than formaldehyde & less
toxic
Sterilize respiratory therapy equipment,

endoscopes & haemodialysis equipment

8. Gases (Ethylene Oxide & Betapropriolactone)

Ethylene Oxide

Bactericidal
Alkylating both proteins & nucleic
acids
Sterilize heart-lung machines,
respirators, sutures, dental

Betapropriolactone (BPL)

equipment, books & clothing

Bactericidal
Fumigation
Sterilize biological products

9. Acids & Alkalis

a. Strong acids & alkalis (e.g. NaOH)
o Bactericidal
o Denaturing proteins
b. Weak acids (e.g. benzoic, propionic & citric acids)
o Bacteriostatic
o Action by organic moiety & low pH
o As food preservatives

10.Dyes
Bacteriostatic-inhibiting growth of bacteria
Modifies nucleic acid
a) Aniline dyes (brilliant green, malachite green & crystal violet)
React with acid groups in the cell
In Lwenstein-Jensens medium: The dye inhibit growth of unwanted
organisms in sputum on culturing M. tuberculosis
As selective agents in culture media
b) Acridine dyes (proflavine, acriflavine, euflavine & aminacrine)
Impair DNA complexes of organisms Killing the cell or destroying
its reproductive capacity

Test to determine efficiency of the chemical agents

No single reliable test available, due to the number of parameters

influencing disinfectant activity
Phenol is taken as the standard (comparison) in tests

Concentration of phenol needed

Phenol coefficient = Concentration of test disinfectant needed

Phenol coefficient =

or

Dilution of test disinfectant needed

Dilution of phenolneeded

Disinfectant more effective than phenol have coefficient >1

Disinfectant less effective than phenol have coefficient <1
1. Rideal-Walker test
Suspensions containing equal number of typhoid bacilli are
submitted to the action of varying concentrations of phenol &
disinfectant to be tested
Does not reflect natural conditions as bacteria & disinfectant react
directly without any organic matter normally present
2. Chick-Martin test
Disinfectant acts in the presence of organic matter such as dried
yeast or faeces

References: Warren Levinson 12th Edition pg 99-102 ; Mims Medical Microbiology

5th Edition pg 529, 530 ; Seniors note. Feel free to add/cross out the stuff you
find important/useless. Thank you.