Escolar Documentos
Profissional Documentos
Cultura Documentos
Syllabus:
•Classification of Disinfectants
Disinfectants generally kills the sensitive vegetative cells but not the
heat resistant endospores. Disinfection is less effective than
Sterilization process
Endospores of bacteria
Mycobacteria
Cysts of protozoa
Vegetative protozoa
Aminacrine hcl
Crystal Violet- X-(CH3)2N, Y-(CH3)2N, Z-(CH3)2.Cl
Brilliant green- X-(C2H5)2N, Y-No group, Z-(CH3)2.Cl
Acridine
Malachite green- X-(CH3)2N, Y-No group, Z-(CH3)2
Detergents
They are used as surface active agents, wetting agents and emulsifiers.
They are classified into four main groups such as anionic, cationic,
nonionic and amphoteric.
Cationic surface active agents are the most antibacterial agents eg.
Cetrimide, benzalkonium chloride.
Anionic surfactants are Soaps, Sodium lauryl sulfate.
Soaps prepared from saturated fatty acids are more effective against
Gram negative bacteria
While prepared from unsaturated fatty acids are effective against G+ve
bacteria and Neisseria group
Non ionic detergents are not ionized and also it donot posses
antimicrobial activity
Amphoteric compounds have properties of both anionic and cationic
detergents
Factors affecting Disinfectant action
The rate and extent of antimicrobial action of chemical disinfectants are influenced
by a number of factors,
1. Concentration of disinfectant
2. Temperature
3. Time of contact
4. pH of the environment
5. Surface tension
•
3. Ditch –plate method:
A solution of antimicrobial substance is carefully run into the
ditch which is prepared in an agar plate.
A loopful of eact test m.o. is then streaked outwards from the
ditch on the agar surface.
M.O. resistant to the antimicrobial grow right upto the ditch
whereas sensitive m.o. shows a zone of inhibition adjacent to
the ditch or centre of plate.
Width of inhibition indicates the Antimicrobial action
4. Phenol coefficient method:
Various disinfectants are tested for its microbicidal property and
they are compared with phenol activity as reference.
The same quantities of micro organisms are added to the
different dilutions of phenol and disinfectants
In UK, Sal. typhi and in USA, Sal.typhi, Staph. Aureus and
Pseud. aeruginosa are used as test micro organisms
The pheno coeffecient test includes,
1. Rideal – Walker test (RW test)
2. Chick – Martin test (CM test)
3. USFDA test
4. The US Association of Official Agricultural Chemists test
(AOAC test)
These tests is only suitable for testing water soluble
disinfectants and exerts the microbicidal action as phenol
In Rideal – Walker test , Rideal – Walker broth (Meat extract, Peptone,
Sodium chloride and Water) is used and Salmonella typhi as sensitive micro
organism.
Different dilutions of test disinfectants and phenol are prepared and 5ml of
each dilution is inoculated with 0.5ml of 24 hrs broth culture of organisms.
All tubes are placed in water bath (20 ºC). Subcultures of each reaction
mixture are taken and transferred to 5 ml sterile broth after 2.5, 5, 7.5 and 10
min
The broth tubes are incubated at 37 ºC for 48 to 72 hours and examined for
the presence or absence of growth.
The Rideal – Walker coeffecient of the test disinfectant is then calculated as
follows,
If a phenol coeffecient or ridal Walkar coeffecient is 1,
then the test disinfectant has the same effectiveness and
if it is less than 1 it means less effective and if it is more
than 1 it means more effective than phenol.
Advantages of Phenol Coeffecient test:
1. Inexpensive and can be performed quickly
2. Have reproducible results
3. They are helpful to eliminate to useless products
Disadvantages of Phenol Coeffecient test:
1. Have only limited information (only one M.O. – S.typhi)
2. Activity of bactericides done at only one concentration with
fixed death time and reaction temperatures
3. In presence of organic matter, the test has no indication of
activity of disinfectants
4. No information about toxicity
5. Sampling errors are large
5. Kelsey – Sykes method (1969):
In this method, several test bacteria are used. They are, Staph.
Aureus, Proteus vulgaris, E. coli and Pseud. aeruginosa.
The final conc. of bacterial cells is about 109/ml. Basic procedure is
outlined in below table.
The samples taken at 8, 18 and 28 minutes are then incubated at 30
to 32 °C and the number of tubes showing growth or the number of
colonies from surface plate culture is recorded.
Result:
No growth occurs in 2 or more of 5 tubes of 18 min samples ie.,
subcultures taken after second incremental addition of bacteria or
There are not more than 5 colonies from the 5 drops on the agar
plate.