STERILIZATION and DISINFECTION

1. Describe the Working of an Autoclave? How does Autoclave destroy

microorganism?
 Microorganisms such as bacteria and spores are killed using heat in an
autoclave. And even heat-resistant endospores from various types of
instruments. This is done by heating the instruments within the device to
temperatures surpassing the boiling point of water. The heat comes from
pressured steam. The steam may reach the high temperatures needed for
sterilizing thanks to the pressurization. Sterilization is accomplished by
exposing liquid handling items to saturated steam at a high temperature
approximately 121°C for the necessary duration.
2. What are the Various Physical Methods of Sterilization? Describe each one of
them?
 Physical methods of sterilization
1. Heat it can be applied to both moisture-sensitive and moisture-
resistant products, for which dry (160–180°C) and moist (121–
134°C) heat sterilization procedures are respectively used. The
effects of heat sterilization occur more rapidly in a fully hydrated
state, as it requires a lower heat input, with low temperature and
less time, under high humidity conditions where the denaturation
and hydrolysis reactions are predominant, rather than in the dry
state where oxidative changes take place.
2. Filtration the process of filtration is unique among sterilization
techniques in that it removes, rather than destroys,
microorganisms. Further, it can prevent the passage of both viable
and nonviable particles and can thus be used for both the
clarification and sterilization of liquids and gases.
3. Radiation- the process of exposing surfaces and objects to different
kinds of radiation for sterilization. The major target for these
radiations is microbial DNA, where damage occurs as a result of
 ionization and free radical production (gamma-rays and electrons)
or excitation (UV light).
3. Explain how to test for efficacy of Disinfectants? Give Examples?
 Manufacturers frequently conduct tests while creating disinfectants, and
the test numbers, as well as the standard contact periods indicated in the
technique, may be printed on the product labels. Standard test procedures
are commonly employed because they are reliable, repeatable, and well-
known. End consumers may also find it valuable to be able to compare
products from different producers by referring to findings from standard
method testing. This test includes:
 1. Koch’s method-
 2. Rideal Walker Method- relies on the estimation of phenol coefficient.
Phenol coefficient of a disinfectant is calculated by dividing the dilution of
test disinfectant by the dilution of phenol that disinfects
underpredetermined conditions.
 3. Chick Martin test - determines the phenol coefficient of the test
disinfectant.
 4. Capacity use dilution test (Kelsey-Sykes test)- determine the appropriate
use dilution of the disinfectants.
4. Classify Disinfectants based on their Mechanism of Actions.
 a) Action on membrane (Ex. Alcohol, detergent)
 (b) Denaturation of cellular proteins (Ex. Alcohol, Phenol)
 (c) Oxidation of essential sulphydryl groups of enzymes (Ex. H2O2,
Halogens)
 (d) Alkylation of amino-, carboxyl- and hydroxyl group (Ex., Formaldehyde)
 (e) Damage to nucleic acids (Formaldehyde) Alcohols Mode of action:
Alcohols dehydrate cells, disrupt membranes and cause coagulation of
protein. Examples: Ethyl alcohol, isopropyl alcohol and methyl alcohol.