Sterilisation and Disinfection

Sterilisation: complete killing of all forms of microorganisms, including bacterial spores

Disinfection: killing or removing of harmful vegetative microorganisms.

Disinfectant: chemical substance used on inanimate objects . Toxic to human

Antiseptic: disinfectant that can be safely used on living tissues.

In the beginning of the 19th century surgical patients very often developed infections since surgery
was not performed under aseptic conditions. Without sterilization and antisepsis, 4 out of 5 operated
patients developed streptococcal or mixed infections. In the first half of the 19th century, surgeons
operated wearing street clothes, without washing their hands and even sutured wounds with
ordinary sewing threads. This is when Louis Pasteur proved that invisible microbes may cause
disease. Joseph Lister then implemented Pasteur’s theory into surgery and came up with the idea of
sterile surgery. He used carbolic acid (phenol) steam spray apparatus to spray into the operating
room.

Sterilization is the complete killing/removal of all microorganisms in a space or object, including

the highly resistant bacterial spores. Sterilization provides the absence of all microorganisms.
Disinfection is the killing of most but not all microorganisms. However, some microorganisms,
especially the bacterial spores may survive. Disinfectants are used only on inanimate objects,
whereas antiseptics are mild disinfectants that are used on living tissues, e.g. skin and mucous
membranes.

STERILIZATION

Steraliszation is the use of physical procedures or chemical agents to destroy all microorganisms
(including bacterial spores) in or on an object.

It is is the full destruction of all microbes, including the more resistant forms such as bacterial
spores (of Bacillus and Clostridium spp.), mycobacteria, viruses, and fungi. It can be performed by
physical, gas vapor or chemical means.

Bacterial spores are the most heat-resistant cells, as a result, their destruction or elimination
guarantees sterility. Before sterilization and disinfection, all organic matters (blood, pus, tissue etc.)
on the objects should be washed away/removed. Even vaporized hydrogen peroxide can fail to
sterilize surgical tools in 3/4 of the cases when the tools were not cleaned enough before
sterilization.

Unlike antibiotics, the sterilization and disinfection have no selective activity. They can damage not
only the microorganisms but also the human cells. Killing depends on numerous factors for
instance the environment and the microorganisms .

To kill endospores, and therefore sterilize a solution or food, very long (1-6 h) boiling is required. It
cannot be obtained home-canned food.
Mechanisms of spore heat resistance

Spores possess very low water content and high calcium content. They possess dipicolinic acid to
chelate calcium and to stabilize DNA by intercalation. Mechanisms of spore heat resistance
involve spore layers such as exosporium outer and inner membranes and cortex,
and DNA-protein complex in the spore.

Method for sterilisations are physical, gas vapour and chemical.

Physical sterilant:

Method concentration or level

Saturated steam 121° C for 15-20 min,

under pressure
(autoclave) pressure of 15 psi = 105

pascals (Pa)
Dry heat 1 h at 171° C; 2 hr at 160° C

Filtration 0.22- to 0.45-μm pore size; HEPA

filters, nanofilters (for viruses)

Ultraviolet Exposure to 254-nm wavelength

radiation

Ionizing radiation Exposure to microwave or gamma

radiation

CHEMICAL STERILANTS

Method concentration or level

Peracetic acid 0.2% 0.2%

Glutaraldehyde(autoclave)

Glutaraldehyde 2%

Physical sterilization

Saturated steam under pressure:

Autoclave is one of the best sterilizing methods. Regimen 121 degrees С/ 15-20 min. at 105 Ра
(pascals) pressure. It denatures proteins/enzymes and nucleic acids (NA), damages membranes.
Decrease penetration of the steam into the load. This is suitable for media, liquids, surgical
instruments, which are not sensitive to heat, humidity and pressure. The disadvantages are its
corrosive for metal, inappropriate for heat-labile materials, leaves
glassware wet.
Physical sterilization- Dry heat

Dry hot air sterilization is not as efficient as the autoclaving because diffusion and penetration of
heat is slow, long sterilization time and very high temperatures are needed, materials can be
damaged by the oxidation during the prolonged heating, and dry heat may stratify in the chamber.

It denatures microbial proteins (enzymes) and NA, membrane damage. Slow diffusion/penetration,
low conductivity compared to autoclaving. This can be applied to glassware, metal, powders; not
plastic/rubber items. It is not corrosive. The disadvantages are that it is slow and damages heat
sensitive materials.

Filtration:
Used to eliminate bacteria and fungi larger than the
filter pore size from air or solutions. The types are depth filters (older: asbestos, glass etc.)
Membrane filters (cellulose acetate) with usually 0.22-0.45 μm pore diameter. Nanofilters with pore
size of 15-40 nm, which can retain viruses. Stops only the microorganisms larger than
the pore diameter. Mycoplasmas and some other small bacteria and viruses go through (except for
the nanofilters). Solution forced through filter with vacuum or pressure. It’s most suitable for
thermolabile solutions, antibiotics, vitamins.

Coronavirus (SARS-CoV-2)
Cloth mask (loose-fitting, reusable) trap droplets released when the wearer talks or coughs or
sneezes. Surgical masks (loose-fitting, disposable masks) protect others from the saliva or
respiratory secretions of the mask wearer. They trap large particles. N95 mask (closely fitting the
face and with valves) offers more protection than a surgical mask does because they can block 95%
of very small particles. They protect the wearer. CDC recommends cloth face masks for the
public and not the surgical and N95 masks needed by health care providers.

Disinfection
The use of physical procedures or chemical agents to eliminate most microorganisms (reducing the
number of pathogenic microorganisms present in a sample). Possible residual organisms present:
bacterial spores and relatively resistant microorganisms (M. tuberculosis, viruses
or fungi) may survive.

ANTISEPSIS:
use of chemical agents on skin and mucosa to inhibit or remove microbes.

Preliminary cleaning:
To repeat preliminary cleaning before disinfection is highly important. Residues from improperly
diluted disinfectants may boost the growth of antibiotic-resistant bacteria. Cleaning can be
performed manually, e.g., by friction or mechanically, e.g., with ultrasonic cleaners or other
methods).

DISINFECTANTS are of high-, intermediate- or low-level activity. High-level disinfectants

eliminate/inactivate completely all microorganisms in or on an instrument except large numbers of
bacterial spores. Intermediate-level disinfectants eliminate all microbial pathogens except bacterial
spores. Low-level disinfectants eliminate vegetative bacteria and lipid-enveloped or medium-size
viruses.
HIGH-LEVEL DISINFECTANTS

They destroy all microorganisms except for spores in high quantity. For semicritical items that come
into contact with either mucosas or nonintact skin.

Used for items for invasive procedures that cannot withstand sterilization (e.g. gastrointestinal
endoscopes, bronchoscopes, surgical instruments with plastic etc).

HIGH-LEVEL DISINFECTANTS
Exemples: moist heat, glutaraldehyde, hydrogen peroxide, peracetic acid, chlorine dioxide and
chlorine compounds, ortho-phthaldehyde

INTERMEDIATE-LEVEL DISINFECTANTS
Destroy vegetative bacteria, including mycobacterium spp. and most viruses and fungi. Used for
semi-critical instruments and devices.Used to clean instruments/surfaces where contamination with
bacterial spores and resistant microorganisms is improbable (endoscopes, laryngoscopes, vaginal
specula, anesthesia breathing circuits).instruments/surfaces where contamination with bacterial
spores and resistant microorganisms is improbable (endoscopes, laryngoscopes, vaginal specula,
anesthesia breathing circuits).

Examples:

- alcohols, iodophors, phenolic compounds, leach and hydrogen peroxide blends.

LOW-LEVEL DISINFECTANTS
They do not kill the spores, mycobacteria and small non-lipid-containing viruses.

Used to clean noncritical instruments that do not penetrate through mucosal surfaces or into sterile
tissues (stethoscopes, blood pressure cuffs, electrocardiogram electrodes, etc.)

Meahek Butt
8207
Group 48