Sterilization and Disinfection

Definition
Disinfection Sterilization

A process that eliminates many or all A process that destroys or eliminates ALL
pathogenic microorganisms, EXCEPT forms of microbial life, including
bacterial spores, on inanimate objects spores
Reduced number of bacteria Performed health-care facilities by physical or
chemical or filtration
methods.
Not sporicidal

The purpose…
• To prevent transmission of microbes to patients
– Direct
– Indirect
Current Threats • Hospital-acquired Infection (Nosocomial)
– Endogenous flora
– Exogenous via Cross-infection
– (El-Demerdash T et al., 2018)
– Environmental surfaces
– Medical devices (critical, semi-critical, non-critical)
Critical Medical • Confer high risk for infection if contaminated
Devices • Enter sterile tissues or vascular sys
• Surgical instrument, cardiac and urinary catheters, implant, ultrasound probe,
diaphragm fitting rings etc.
Semi-critical MDs • In contact with mucous membranes or non-intact skin
• Respiratory therapy and anesthesia equipment, some endoscopes, laryngoscope
blades, oesophageal manometry probes etc.

Non-critical MDs • In contact with skin but not mucous membrane

• Bedrail, bedpan, blood pressure cuffs, crutches, computers, wheelchairs

1
Frances
 Sterilization and Disinfection

Current Threats • Bioterrorism: Anthrax

• Emerging pathogens: Candida auris, Carbapenem-resistant Enterobacteriaceae
• Re-emerging: Zika, Ebola, MDR pathogens
Microbiological
Disinfectant
Hierarchy

Differences / • Security level (risk of spreading infection)

Factors to • Inactivation of all forms of microorganisms (incl. spores)
Consider: • Efficacy of sterilant
• Duration of process
• Prevent deposition of new pathogens post processing
What are the
foresee-able
difficulties /
challenges with
cleaning medical
devices?

Other Factors to Consider:

• Prior cleaning (manual, mechanical, enzymatic)
• Organic or inorganic load (reduce interference)(Nandy et al., 2018)
• Type of microbial contamination (to decontaminate: safe to handle, use or discard)
• Level of microbial contamination (low / intermediate / high)
2
Frances
 Sterilization and Disinfection
- Give examples
• Concentration of germicide
• Exposure time of germicide

Difference between antiseptic and disinfectants?

5
Factors to • Physical nature of object (crevices, hinges)
consider: • Presence of biofilms
• Physical • Relative humidity (EtO)
• Chemical
• Gaseous Chemicals • Types of chemical (pH, causticity)
• Ultrasonic • Site of administration
• Dosage (conc. , time)
• Functions
– disruption of lipid cell membrane
– modification of proteins
– modification of DNA
Disinfectants (1) • These are not interchangeable, and incorrect concentrations
and inappropriate disinfectants can result in excessive costs.
• causes occupational diseases among cleaning personnel
(e.g., formaldehyde, glutaraldehyde, and chlorine)
• PPE (e.g., gloves and proper ventilation)
Disinfectants (2) • Asthma and reactive airway diseases can occur in sensitized
persons exposed to any airborne chemical, including
germicides.
• Preferred method of control is elimination of the chemical
(through engineering controls orsubstitution) or relocation of
the worker.
Disinfectants • used alone or in combinations (e.g., hydrogen peroxide and
peracetic acid) in the health-care setting.
• include alcohols, chlorine and chlorine compounds,
formaldehyde, glutaraldehyde, ortho-phthalaldehyde,
hydrogen peroxide, iodophors, peracetic acid, phenolics, and
quaternary
ammonium compounds.
Classification of • Consistency
Disinfectants – Liquid (alcohol, phenols)
– Gaseous (formaldehyde vapour)
• Spectrum of activity
– High
– Intermediate
– low
Properties of ideal • Wide spectrum activity
disinfectants: • Destroy microbes within practical duration
• Effective contact and wettable
• Active in any pH
• Stable with long shelf life
• High penetrating power

3
Frances
 Sterilization and Disinfection
• Cheap, available, accessible
Chemicals: • Alcohol (D)
Disruption of cell • Detergents (D)
membrane • Phenols (D)

Chemicals: • Formaldehyde (D)

Modification of • Glutaraldehyde (D)
Proteins • Iodine (D)
• Chlorine (D)
• Heavy metals (D)
• Acids and Alkalis (D)
Chemicals: • Hydrogen peroxide (D)
Modification of • Crystal violet (D)
Nucleic acids • Malachite green (D)
• Ethylene Oxide (S)
• 73-80 C steam+formaldehye (S)
E.g. 2%
glutaraldehyde

High level • Semi-critical devices (gastrointestinal

Disinfection endoscopes, endotracheal tubes, anaethesia breathing circuits
etc.)

• Immediate, compatible enzymatic cleaner

• Discard enzy. cleaner once used
• Heat sterilization if heat stable.
4
Frances
 Sterilization and Disinfection
• Appropriate pressure for Flush and correct size for brush all
accessible channels

Low level • Non-critical equipment (eg. arthroscope, laparoscope,

Disinfection cystoscope)
• Bedrails, bedpans, over-the-bed-table, blood pressure cuff

How do we know • effective dilution

the • duration for effective disinfection
disinfectant is • periodical monitoring
working?
Methods to assess:
• Koch’s method: B anthracis on silk thread, disinfectant,
washed, transferred to solid media to observe growth

• In-use test: Estimate number of living organism in a vessel

of disinfectant currently in use. 1:10 dilution and place 10
drops on each of 2 nutrient agar plates;
37C, 3 days and room temp., 1/52. If ≥5 drops on either
plate has growth, FAIL
Enzymes • Enzymes, usually proteases, sometimes are added to neutral
pH solutions to assist in removing organic material (blood,
pus).
• Lipases (enzymes active on fats)
• Amylases (enzymes active on starches).
• Enzymatic cleaners are not disinfectants
• Proteinaceous enzymes can be inactivated by germicides.
Heat: • 62C for 30 min with rapid cooling
Pasteurization • ‘Flash pasteurization’ 72 C for 15 sec
• kill vegetative milk-borne pathogens (e.g., Mycobacterium
bovis, Salmonella, Streptococcus, Listeria, and Brucella)
• Not used to sterile milk

5
Frances
 Sterilization and Disinfection
Various methods
of
Sterilization

Heat: Moist Heat • Moist heat (boiling or autoclaving)

– Sterilised at lower temp as water helps disrupt H-bonds
– Bacterial spores resistant to boiling (100C)
– Autoclaving: 121C for 15-20 min
Autoclave • 121-134C
• Steamed under pressure
• Saturated steam destroy microorganisms
• Most dependable, accessible
• For metal, glass, NOT rubber, plastics or items easily damaged by high temp.

6
Frances
 Sterilization and Disinfection

Heat: Dry Heat • 180 C for 2 hrs

• kills highly heat-resistant Clostridium botulinum spores
• Glassware
• Less freq use cf autoclaving
Radiation • Radiation: type, dosage (conc. , time)
• Radiation (UV and X-ray) kill by damaging DNA
– Gamma-photons (S)
– Beta-Electron (S)
– UV 250-260nm (D)
– X-ray (D)

Radiation, Gas

7
Frances
 Sterilization and Disinfection

Why 250-260nm?
• Maximum absorption by purine and pyrimidine bases of DNA
• Formation of thymine dimers
• Addition of hydroxyl grps to bases
• DNA replication inhibited
• Unless against bacterial spores, requires 10X.

Filtration • Sterilise liquid if pore size is small enough (0.22 micro) which retains all bacteria
and spores (size and electrostatic)
• For heat-sensitive liquids.
– Intravenous fluids (antibiotics, drugs,…)
• To eliminate heat-resistant endotoxin in cell walls of dead Gram-ve bac
– Pyrogen-free

Ultrasonication • Users of ultrasonic cleaners should be aware that the cleaning fluid could result in
endotoxin contamination of surgical instruments, which could cause severe
inflammatory reactions (Richburg et al., 1986)
Biological • Selected BI with spores Bacillus atrophaeus, Bacillus anthracis
indicators • Gas sterilization: B. subtilis var. niger, B. subtilis var. golbigii
• Sterilization process monitoring

8
Frances
 Sterilization and Disinfection
Disposal

~End😊

9
Frances