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DEFINITION - DISINFECTION
Process of elimination of most pathogenic organisms excluding bacterial spores on inanimate objects. Sterilization and Disinfection are NOT synonymous.
Non-toxic/ Non-allergic/
Non-corrosive/Non-irritant
organic matter
Considerations of Disinfection
Factors influencing the effectiveness of a disinfectant:
As disinfectants are known to loose their action on long standing & in the presence of organic matter their efficacy must be tested periodically.
BASIC PRINCIPLE
All these tests use the same principle in evaluating
contact with the disinfectant, their viability is tested by sub-culturing them on media prepared without disinfectant The disinfectant carried over to the sub-cultures should therefore neutralized either by diluting them to subinhibitory conc. Or by adding a substance that inactivates it.
1/10 dilution for phenols, aldehydes 0.5% Na.thiosulphate for chlorine comps, iodophores 1% Na. bisulphate for HCHO and glutaraldehude
KOCH'S METHOD
Spores of Bacillus anthraces were dried on silk thread and were subjected to action of disinfectants. Later, it was washed and transferred to solid medium.
Measures the lowest concentration of the disinfectant that inhibits the growth of known strain of organism.
Disadvantages: i. No. of organisms considered-too low ii. Time of exposure- too long iii. No sufficient organic matter
Inoculate streaks of different bacterial suspensions onto each plate of NA. Incubate for 37 C-48hrs
The lowest concentration that entirely prevents the growth of the indicator bacteria is the MIC of that disinfectant in that particular medium.
Bactericidal potency is assessed by measuring the rate of killing a selected range of bacteria under specified conditions.
B.cidal power of a particular disinfectant as compared with that of pure PHENOL. Minimum time of 10min. Usually employed cultures are S.typhi& St.aureus. Phenol coefficient is considered to be 1.
Determine MIC of disinfectant for S.typhi Prepare 5 graded concentrations in DW lowest conc.<MIC
Determine MIC of phenol for S.typhi Prepare dilutions of 1/95, 1/100, 1/105, 1/110, 1/115
Take 5ml into sterile test tube Add 0.2ml of 24hr broth of S.typhi Shake well
At regular intervals (say 5min) remove a large loopful from each mixture& transfer to a tube of 5ml broth
Inoculate broth for 48hrs
Calculate phenol coefficient by dividing the figures indicating the dilution of test that shows growth after exposure for 5min to that of no growth after long exposure-10min
Disadvantages: 1. Ineffective in the presence of organic matter. 2. Test is not reproducible 3. Can be done only for phenolic disinfectants 4. S. typhi may not be appropriate 5. The time allowed for disinfection is short
CHICK-MARTIN TEST
Modification of Rideal Walker Method
GARRODS TEST
Modification of Chick-Martin Test. Contains yeast instead of feces. 5g of dry yeast is mixed with 100ml DW.
S.typhi
disinfectant in actual use. The disinfectant that is already in use is diluted 1/10 by mixing 1 ml of the disinfectant with 9 ml of sterile nutrient broth. Ten drops of the diluted disinfectant (each 0.02 ml) is placed on two nutrient agar plates One plate is incubated at 37oC for 3 days while the other is held at room temperature for 7 days The number of drops that yielded growth is counted after incubation If there growth in more than five drops on either plate, it represents failure of disinfectant.
9ml of quarter strength Ringer soln 9ml isotonic soln that inactivates it
Prepare a suspension Transfer 10 small drops onto separate areas of the surface of each of two dried agar plates Incubate at 37 C-48hr Incubate at room temp
to determine concentrations of disinfectant that will be effective in clean and dirty conditions. Method:
Effective in clean and dirty conditions. The dilutions of the disinfectant are made in hard water for clean
conditions and in yeast suspension for dirty conditions The contact time of disinfectant and test organism is 8 min. Test organism alone or with yeast is added at 0, 10 and 20 minutes interval Depending on the type of disinfectant, a single test organism is selected from S. aureus, P. aeruginosa, P. vulgaris and E. coli.
Test disinfectant + standard hard water solution Add suspensions of test bacterium After each addition of organism, portion of the mixture are removed
The disinfectant is evaluated on its ability to kill microorganisms or lack of it and the result is reported as a pass or a fail and not as a coefficient. Disadvantage: Rather complicated.
STABILITY TESTS
It measures the stability & long-term effectiveness of the diluted disinfectant in clean and dirty medium. Used to supplement the information obtained from capacity use dilution tests.
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