deepika m

Decontamination
the

process of removal of contaminating pathogenic microorganisms from the articles by a process of sterilization or disinfection use of physical or chemical means to remove, inactivate, or destroy living organisms on a surface so that the organisms are no longer infectious.

Decontamination steps

CLEANING

STERLIZATION

DISINFECTION

STEAM

DRY

CHEMICAL

HIGH LEVEL

INTERMEDIATE LEVEL

LOW LEVEL

CLASSIFICATION OF INSTRUMENTS DEPINDING UPON ITS USE

CRITICAL

penetrate soft tissue or bone, enter the bloodstream or normally sterile tissue.

forceps, scalpels, bone chisels, scalers and surgical burs

steam under pressure (autoclaving), dry heat, or heat/chemical vapor

SEMICRITICAL

contact mucous membrane or non-intact skin

mirrors,impressio n trays,amalgam condensers,Endos copes,Anesthesia equipment

High level disinfection

NONCRITICAL

objects that touch only intact skin

x-ray heads, blood pressure cuffs and pulse oximeters

Low level disinfection

DISINFECTION
Process

of elimination of most pathogenic microorganisms (excluding bacterial spores) on inanimate objects by :-physical methods chemical methods

Achieved

STERLIZATION
process where all the living microorganisms, including bacterial spores are killed. Achieved by :-physical chemical physiochemical methods

Classification of disinfectants:
Based on consistency Liquid (E.g., Alcohols, Phenols) Gaseous (Formaldehyde vapor, Ethylene oxide) Based on spectrum of activity High level Intermediate level Low level Based on mechanism of action Action on membrane (E.g., Alcohol, detergent) Denaturation of cellular proteins (E.g., Alcohol, Phenol) Oxidation of essential sulphydryl groups of enzymes (E.g., H2O2, Halogens) Alkylation of amino-, carboxyl- and hydroxyl group (E.g., Ethylene Oxide, Formaldehyde) Damage to nucleic acids (Ethylene Oxide, Formaldehyde)

Chemical methods of disinfection

chemicals are known as disinfectants.

Properties : Should have wide spectrum of activity Should be able to destroy microbes within practical period of time Should be active in the presence of organic matter Should make effective contact and be wettable Should be active in any pH Should be stable Should have long shelf life, high penetrating power Should be non-toxic, non-allergenic, non-irritative or noncorrosive Should not have bad odour Should not leave non-volatile residue or stain Should not be expensive and must be available easily

High Level Disinfection (HLD)

There are three types of HLD: Disinfection by boiling

Moist heat at 70-100C Chemical disinfection by:-Ethylene Oxide, Gluteraldehyde, Formaldehyde

Intermediate level disinfection by:Phenolics halogens

Low level disinfection by:Alcohols, quaternary ammonium compounds

Mechanism of action
ALCOHOLS: Mode of action: Alcohols dehydrate cells, disrupt membranes and cause coagulation of protein.

Examples: 70% Ethyl alcohol, isopropyl alcohol and methyl alcohol Uses:disinfect surfaces clinical thermometers methyl alc kills spores Disadvantages: Skin irritant, volatile (evaporates rapidly), inflammable

ALDEHYDES: Mode of action: Acts through alkylation of amino-, carboxyl- or hydroxyl group, and probably damages nucleic acids. It kills all microorganisms, including spores. Examples: Formaldehyde, Gluteraldehyde Uses :40% Formaldehyde (formalin) is used for surface disinfection and fumigation of rooms, chambers, operation theatres, biological safety cabinets, wards, sick rooms etc Disadv : Vapors are irritating, has poor penetration, leaves non-volatile residue, activity is reduced in the presence of protein

PHENOLS act as disinfectants at high concentration and as antiseptics at low concentrations used to prevent infection of surgical wound Disadv:toxic, corrosive and skin irritant. HALOGENS Iodophores permit slow release and reduce the irritation of the antiseptic. For hand washing iodophores are diluted in 50% alcohol. 10% Povidone Iodine is used undiluted in pre and postoperative skin disinfection Disadv: Rapidly inactivated in the presence of organicmatter. Iodine is corrosive and staining.

HYDROGEN PEROXIDE: Mode of action: acts on the microorganisms through its release of nascent oxygen.Produces hydroxyl-free radical that damages proteins and DNA. Application: It is used at 6% concentration to decontaminate the instrumentes for 20 30 min, equipments such as ventilators. 3% Hydrogen Peroxide Solution is used for skin disinfection and deodorising wounds and ulcers. Strong solutions are sporicidal.

HLD BY BOILING
best achieved by moist heat such as boiling in water (100C for one minute holding time) kills all organisms except for a few bacterial spores. note that boiling equipment items in water will not achieve sterilization.

STERLIZATION METHODS
STERLIZATION

PHYSICAL

CHEMICAL

PHYSIOCHEM ICAL

SUNLIGHT

HEAT

VIBRATION

RADIATION

FILTRTION

RED HEAT

FLAMING

DRY HEAT

INCENERATION

HOT AIR OVEN

HEAT

INFRARED

BELOW 100 DEGREE

MOIST HEAT

AT 100

ABOVE 100

DRY HEAT:
Red

heat: Articles such as bacteriological loops, straight wires, tips of forceps and searing spatulas holding them in Bunsen flame become red hot. This is a simple method for effective sterilization

Flaming: Passing the article over a Bunsen flame, but not heating it to redness.

Articles such as scalpels, mouth of test tubes, flasks, glass slides and cover slips are passed through the flame a few times. Most vegetative cells are killed, there is no guarantee that spores too would die on such short exposure.
This method too is limited to those articles that can be exposed to flame. Cracking of the glassware may occur.

Incineration: Method of destroying contaminated material by burning them in incinerator.

Articles such as soiled dressings; animal carcasses, pathological material and bedding etc should be subjected to incineration. This technique results in the loss of the article, hence is suitable only for those articles that have to be disposed.

Burning of polystyrene materials emits dense smoke, and hence they should not be incinerated.

Hot air oven: Introduced by Louis Pasteur.

Articles to be sterilized are exposed to high temperature (160C) for duration of one hour in an electrically heated oven. sterilized: Metallic instruments (like forceps, scalpels, scissors), glasswares (such as petri-dishes, pipettes, flasks, all-glass syringes), swabs, oils, grease, petroleum jelly and some pharmaceutical products.

Articles

Sterilization process: Articles to be sterilized must be perfectly dry before placing them inside to avoid breakage. Articles must be placed at sufficient distance so as to allow free circulation of air in between Sterlization cycle 60 minutes at 160C 40 minutes at 170C 20 minutes at 180C. Increasing temperature by 10 degrees shortens the sterilizing time by 50 percent. The hot air oven must not be opened until the temperature inside has fallen below 60C to prevent breakage of glasswares.

Advantages: It is an effective method of sterilization of heat stable articles. The articles remain dry after sterilization. This is the only method of sterilizing oils and powders. Disadvantages: Since air is poor conductor of heat, hot air has poor penetration. Cotton wool and paper may get slightly charred. Glasses may become smoky. Takes longer time compared to autoclave.

Infra red rays: Brings sterilization by generation of heat

Articles to be sterilized are placed in a moving conveyer belt and passed through a tunnel that is heated by infrared radiators to a temperature of 180C. The articles are exposed to that temperature for a period of 7.5 minutes. Articles sterilized included metallic instruments and glassware. It is mainly used in central sterile supply department

Not applicable in diagnostic laboratory

Moist heat
Destroys

through irreversible coagulation of proteins Moist heat includes Boiling Pasteurization Pressurized steam

Boiling (100 C) Destroys most microorganisms and viruses Not effective means of sterilization Does not destroy endospores Pasteurization(below 100deg) Pasteur developed to avoid spoilage of wine Does not sterilize but significantly reduces organisms Used to increase shelf life of food Most protocols employ HTST method,Heated to 72C and held for 15 seconds Other protocol UHT(ultra high temp)Heated to 140C 150C, held for several seconds then rapidly cooled

Pressurized steam(above 100deg) Autoclave used to sterilize using pressurized steam Heated wate steam increased pressure Preferred method of sterilization Achieves sterilization at 121C and 15lbs in 15 minutes Effective against endospores Flash autoclaving sterilizes at 135C and 15lbs in 3 minutes Prions destroyed at 132C and 15lbs for 4.5 hours

Other Physical Methods of Control

Heat sensitive materials require other methods of microbial control
Filtration Irradiation High-pressure

treatment

Filtration Membrane filtration used to remove microbes from fluids and air Liquid filtration Used for heat sensitive fluids Membrane filters allow liquids to flow through Traps microbes on filter Depth filters trap microbes using electrical charge

Filtration of air High efficiency particulate air (HEPA) filter remove nearly all microbes from air Filter has 0.3m pores to trap organisms

Radiation Electromagnetic radiation Energy released from waves Based on wavelength and frequency Shorter wavelength, higher frequency = more energy Range of wavelength is electromagnetic spectrum Radiation can be ionizing or non-ionizing

Ionizing radiation Radiation able to strip electrons from atoms Three sources Gamma radiation X-rays Electron accelerators Causes damage to DNA and potentially to plasma membrane Used to sterilize heat resistant materials Medical equipment, surgical supplies, medications Some endospores can be resistant

Non-ionizing radiation Ultraviolet radiation Only type to destroy microbes directly Damages DNA Causes thymine dimers Used to destroy microbes in air, drinking water and surfaces Limitation Poor penetrating power Thin films or coverings can limit effect

High pressure processing Used in pasteurization of commercial foods

Does

not use high temperatures high pressureUp to 130,000 psi

Employs

Destroys

microbes by denaturing proteins and altering cell membrane permeability

Sterlization control Three methods exist to check the efficacy of sterilization process Physical Chemical Biological.

 Physical: Temperature

chart recorder and

thermocouple.
Chemical:

Brownes tube No.3 (green spot, color changes from red to green)

Biological:

10^6 spores of Bacillus subtilis var niger or Clostridium tetani on paper strips are placed inside envelopes and then placed inside the hot air oven.Upon completion of sterilization cycle, the strips are removed and inoculated into thioglycollate broth or cooked meat medium and incubated at 37C for 3-5 days. Proper sterilization should kill the spores and there should not be any growth.

Summary
Disinfection

and sterilization guidelines must be followed to prevent exposure to pathogens that may lead to infection of sterile products for use in patient care depends not only on the effectiveness of the sterilization process but also on cleaning, disassembling and packaging of the device, loading and monitoring the sterilizer

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