STERILIZATION INTRODUCTION STERILIZATION This term refers to the process that eliminate (removes) or kills all forms of life,

including transmissible agents (such as fungi, bacteria, virus, sporeforms, etc) present on a surface, contained in a fluid, in medication, or in a compound such as biological culture media. Destruction of microorganisms means they cannot be revived to multiply even when transferred from the sterilized product to an ideal growth medium. A sterile item is one that is absolutely free of microbes, including endospores and viruses. There are two main methods of sterilization, namely, physical and chemical methods of sterilization.

PHYSICAL STERILIZATION Moist heat sterilization The presence of moisture increases the effectiveness of heat in accomplishing the destruction of microorganisms. Moist heat has greater penetrating power and coagulates protoplasm at lower temperatures (e.g. 70 degrees Celsius). Destruction of these microorganisms is also achieved within a shorter time frame as compared with dry heat. An example is the autoclave. Boiling in water is another method of moist heat sterilization. Boiling in water for ten minutes at 100 degrees Celsius will kill most vegetative bacteria and inactivate viruses except endospores, including those of the food poisoning bacteria Clostridium perfringens and C.botulinum, which can survive many hours of boiling. Unfortunately boiling is ineffective against prions and many bacterial and fungal spores; therefore boiling is unsuitable for sterilization unless adequate time is allowed. However, since boiling does kill most vegetative microbes and viruses, it is useful for reducing viable levels if no better method is available. CHEMICAL STERILIZATION Many of the substances used in preparing culture media are too heat-labile to be sterilized by autoclaving. For such substances, reliable methods of filtration and chemical sterilization will be more useful. The essential requirement for a chemical sterilizing agent is that, it should be volatile and toxic [pertaining to microorganisms], so that it can be eliminated readily from the object sterilized after treatment. Ethylene oxide gas (EO or EtO) This is commonly used to sterilize objects sensitive to temperatures greater than 60 C and / or radiation such as plastics, optics and electrics. Ethylene oxide penetrates well, moving through paper, cloth, and some plastic films and is highly effective. EtO can kill all known viruses, bacteria and fungi, including bacterial spores and is compatible with most materials (e.g. of medical devices), even when repeatedly applied. However, it is highly flammable, toxic and carcinogenic. This method has drawbacks

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because of air contamination as a result of release of CFCs and/or large amounts of EtO residues which are hazardous, flammable and has storage issues calling for special handling, operator exposure risk and training costs Ethylene oxide is both explosive and toxic for humans so when handling it care must be taken, because of this ethylene oxide sterilization is not often used in laboratory procedure. In culture media chloroform is often used and the proportion of 2.0% of chloroform is added and can later be removed at heating at 57 degrees Celsius. Hypochlorite or Chlorine bleach This is another accepted liquid sterilizing agent. Household bleach consists of 5.25% sodium hypochlorite. It is usually diluted to 1/10 immediately before use; however to kill Mycobacterium tuberculosis it should be diluted only 1/5, and 1/2.5 (1 part bleach and 1.5 parts water) to inactivate prions. Bleach will kill many, but not all spores. It is also highly corrosive. Bleach decomposes over time when exposed to air. Some examples of chlorine bleach are parazone, and chlorine tablet. Glutaraldehyde this method is used to sterilized equipments destroyed by heat endoscopes [contain lenses]. This liquid sterilizing agent is accepted providing that the immersion time is sufficiently long. Using Glutaraldehyde it will take up to22 hours to kill all spores. It can also be used as a fixative [a solution used to preserve or harden fresh tissue of cell specimens for microscopic examination]. Glutaraldehyde is expensive. Formaldehyde is a solution ( used as fixatives at a concentration of 10%)accepted as a liquid sterilizing agent, provided that the immersion time is sufficiently long. To kill all spores in a clear liquid can take longer with formaldehyde. The presence of solid particles may lengthen the required period or render the treatment ineffective. Sterilization of blocks of tissue can take much longer, due to the time required for the fixative to penetrate. Formaldehyde is volatile, and toxic by both skin contact and inhalation. Formaldehyde is less expensive. It is also used as a gaseous sterilizing agent; in this case, it is prepared on-site by depolymerization of solid paraformaldehyde. Many vaccines, such as the original Salk polio vaccine, are sterilized with formaldehyde. Hydrogen peroxide is another chemical sterilizing agent. Hydrogen peroxide is strong oxidant and these oxidizing properties allow it to destroy a wide range of pathogens and it is used to sterilize heat or temperature sensitive articles such as rigid endoscopes. In medical sterilization hydrogen peroxide is used at higher concentrations, ranging from around 35% up to 90%. It has a short cycle of 28 minutes. Hydrogen peroxide is primary irritant and the contact of the liquid solution with skin will cause bleaching or ulceration depending on the concentration and contact time. The vapor is also hazardous with the target organs being the eyes and respiratory system. Hydrogen peroxide sterilizers have their drawbacks such bleaching and tearing linen if not sluiced well, corrosive, skin irritations and rusting metal. Since hydrogen peroxide is a strong oxidant, there are material compatibility issues and users should consult the manufacturer of the article to be sterilized to ensure that it is compatible with this method of sterilization. References concise medical dictionary, 6th edition[2002],oxford university press. WHO - Infection Control Guidelines for Transmissible Spongiform Encephalopathies. Retrieved Jul 10, 2010.

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CDC-Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008 Prions, p. 100. Retrieved July 10, 2010. Basic medical microbiology, 2nd edition, Robert F. Boyd and Bryan G. Hoerl, 1981, little brown and company (inc), page 133163. Laboratory safety, G. L. Williams, 2007, page 9-13 Microbiology a human perspective, 4th edition, Eugene w. nester, Denise G. Anderson, C. Evans Roberts, jr. , Nancy n. Pearsall, Martha T. Nester, 2004, McGraw-Hill Company, Inc, page 109-115.

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