Decontamination, Disinfection, and Sterilization

Decontamination,
Sterilization, and
Disinfection
Riel Anthony T. Oli, RPh,

MSPharm
PASMETH/PhBBA 5/29/18
Definitions
 Decontamination – A process of cleansing an object or a

substance.
 Decontamination render an area, device, item, or
material safe to handle, that is reasonably free from a
risk of disease transmission.
Biosafety process
 Decontaminate equipment and work surfaces routinely

 With proper disinfectant for agent
 After spills or contamination
 When work is finished
 Decontaminate equipment before removal from lab
 Decontaminate potentially infectious materials before
disposal or reuse
Different Methods of Decontamination
 Sterilization – act or process, physical or chemical, that

destroys or eliminates all forms of life, especially
microorganisms.
 Sterilization is absolute.
 Disinfection – generally less lethal process than

sterilization. It is the elimination of nearly all
recognized pathogenic microorganisms but not
necessarily all microbial forms.
 Chemical
 Thermal
 Radiation
 Filtration
Chemical Disinfectants
 Halogens
 Aldehydes (Formaldehyde, gluteraldehyde)
 Phenolics
 Alcohols
 Acids & Alkalis
 Oxidizing agents (Hydrogen peroxide)
 Quaternary Ammonium compounds (Benzalkonium Cl)
Biguanides
PASMETH/PhBBA (Chlorhexidine) 5/29/18
Factors Affecting Disinfection
 Number of microorganisms
 Location of microorganisms
 Innate resistance to the disinfectant
 Concentration and potency of the disinfectant
 Physical and chemical factors
 Presence of organic matter
 Appropriate contact time between disinfectant and the
contaminant
Biofilms
PASMETH/PhBBA 5/29/18
Environmental Factors
 Dried Spills – may limit contact time between the

disinfectant and the target organism.
 Dirt, grease and oils – all can protect the organisms.
Product Factors
 Age of the product / solution

 Method of application (spray vs wipe)
 Rate of application
 Storage conditions (Opaque vs clear containers)
Resistance to disinfectants
Resistant Prion
Bacterial spores
Coccida
Mycobacteria
Nonlipid or small viruses

Fungi
Vegetative bacteria
Susceptible
PASMETH/PhBBA
Lipid
5/29/18
or medium-sized virus (HIV, herpes,

Resistance to Disinfectants
 Intrinsic resistance
 Gram (-) > Gram (+)
 Spores surrounded by impermeable coat
 Acquired resistance
 Genetic changes
 Use of disinfectants at sub-lethal concentrations
High Level Disinfection - Sporocides
 Kills all microorganisms except high numbers of

bacterial spores
 Requires 5 – 10 minutes exposure
 E.g. aldehydes, hydrogen peroxide, paracetic acid
Intermediate Level Disinfection -
Tuberculocides
 Kills M. tuberculosis var. Bovis and all vegetative
bacteria, fungi, and most viruses
 Require minimum 20 minutes exposure
 E.g. phenolics, iodophores, chlorides, alcohols
Low Level Disinfection – Hospital
Germicides used for Housekeeping
 Kills most vegetative bacterial and some fungi, but not
M. tuberculosis var Bovis
 Require minimum 20 minutes exposure
 E.g. quaternary ammonium compounds
Organic Load
 Blood, sputum, milk, bedding, feed, manure/dung

 Physically protect microorganisms from contact with
disinfectants
 Can neutralize many germicides
Disinfectant Log Reduction Listeria
TSB serum milk
Ethanol 3 1 1
Sodium hypochlorite 5 1 1
Glutaraldehyde 3 3 1
Sodium dichloroisocyanurate 4 1 4
Surface topography
 Uneven, cracked, or pitted surfaces can hide

microorganisms and are difficult to disinfect
Surface Organisms/ 100sq cm
Wood 22,500
Concrete 12,500
Brick 76,500
plastic 13,900
metal
PASMETH/PhBBA 100 5/29/18
Contact Time
 Disinfectants should be effective in a short contact time

 Manufacturer’s recommended contact time may be
unrealistic
 Method of application:
 Surface disinfection – 1 minute
 Instrument soak – 20 minutes
 Loss by evaporation
 Extending contact time does not increase effectiveness in all
cases
Temperature
 Elevated temperatures may enhance action

 Manufacturer’s recommended in-use temp may be hard
to achieve
 Elevated temp may enhance evaporation, thus reducing
contact time
Relative Humidity
 Can influence the activity of some disinfectants

 Fumigation using formaldehyde gas requires an Rh
preferably in excess of 70%
Water hardness
 Germicides may be less effective when diluted in hard

water; varies geographically
Disinfectant Log Reduction S. aureus

50 ppm 200 ppm CaCO3
Ethanol 5 5
Sodium hypochlorite 5 5
Phenolic 4 1
Peroxygens
PASMETH/PhBBA 4 5/29/18 5
The autoclave
Autoclave
 Strong, pressurized, and steam heated vessel

 Settings:
 121 deg C (250F) 15 PSI x 15 minutes
 132 dec C (270F)
 Most dependable system available for decontamination
Autoclave Hazards
 Explosive breakage of glass vessels during opening and

unloading
 Burns arising from careless handling of vessels
containing boiling liquids
 Burns resulting from physical contact
What can be autoclaved?
 Pathogenic plant matter

 Culture and stocks of infectious agents
 Contaminated solids
 Discarded live and attenuated vaccines
 rDNA
 Animal tissues
 Animal cage waste
What should not be autoclaved?
Autoclave Bags and Other Containers
 Polypropylene bags are used to contain materials during

decontamination cycles within autoclaves
 Bags should be loosely closed
 Avoid dry packages
 Loosen caps or cover with foil
Loading
 Steam must contact all items

 Do not overload chamber
 Do not overload the bags
 Keep bags loosely gathered
Cleaning
 Clean trap at least once a week

 Clean surrounding area after every use
 Remove broken glass
 Clean up overflowing liquids immediately
Area Decontamination
 Vapor Phase Hydrogen Peroxide

 Formaldehyde vapor
 Chlorine Dioxide
Key Messages
 No disinfectant is ideal
 There are number of factors that determines how
effective a particular disinfectant is.
 Microorganisms have various innate resistance to
disinfectants.
Key Messages
 Autoclaves use wet heat with appropriate time,

pressure, and temperature
 Wet heat is more effective than dry heat
 Validation of equipment is NECESSARY.

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Decontamination,

Riel Anthony T. Oli, RPh,

 Decontamination – A process of cleansing an object or a

 Decontaminate equipment and work surfaces routinely

 Sterilization – act or process, physical or chemical, that

 Disinfection – generally less lethal process than

 Dried Spills – may limit contact time between the

 Dirt, grease and oils – all can protect the organisms.

 Age of the product / solution

Nonlipid or small viruses

or medium-sized virus (HIV, herpes,

 Kills all microorganisms except high numbers of

 Blood, sputum, milk, bedding, feed, manure/dung

 Uneven, cracked, or pitted surfaces can hide

Surface Organisms/ 100sq cm

 Disinfectants should be effective in a short contact time

 Elevated temperatures may enhance action

 Can influence the activity of some disinfectants

 Germicides may be less effective when diluted in hard

Disinfectant Log Reduction S. aureus

 Strong, pressurized, and steam heated vessel

 Explosive breakage of glass vessels during opening and

 Pathogenic plant matter

 Polypropylene bags are used to contain materials during

 Steam must contact all items

 Clean trap at least once a week

 Vapor Phase Hydrogen Peroxide

 Autoclaves use wet heat with appropriate time,

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