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Definitions:
Sterilization: Killing or removing all forms of microorganisms (including bacterial spores) in a material or an object. Autoclaving is the most commonly used method of sterilization.
Disinfection: is defined as killing most, but not all microorganisms in a location, however some bacterial spores may still remain behind. It involves reducing the number of pathogenic microorganisms to the point where they no longer cause disease.
May use physical or chemical methods.
Phenol coefficient used to evaluate ability of disinfectant to kill MO (ratio of conc. Of phenol to disinfectant to cause same amount of killing.)
Definitions
Disinfectant:
Applied to inanimate objects. Antiseptic: Applied to living tissue (antisepsis). Degerming: Mechanical removal of most microbes in a limited area. Example: Alcohol swab on skin. Sanitization: Use of chemical agent on food-handling equipment to meet public health standards and minimize chances of disease transmission. e.g: Hot soap & water.
Definitions
Sepsis: Comes from Greek for decay or putrid. Indicates bacterial contamination. Asepsis: Absence of significant contamination. Aseptic techniques are used to prevent contamination of surgical instruments, medical personnel, and the patient during surgery. Aseptic techniques are also used to prevent bacterial contamination in food industry.
An agent that kills bacteria. Most do not kill endospores. Viricide: An agent that inactivates viruses. Fungicide: An agent that kills fungi. Sporicide: An agent that kills bacterial endospores of fungal spores.
Disinfectant Performance
Is dependent on Disinfectant concentrations Is dependent on length (time) of administration Is dependent on temperature during administration Microbe type (e.g., mycobacteria, spores, and certain viruses can be very resistant to disinfectionin general vegetative cells in log phase are easiest to kill) Substrate effects (e.g., high organic content interferes with disinfection. It is easier (and faster) to kill fewer microbes than many microbes
Chemical Antimicrobials
Agent
Mechanisms of Action
Comments
Alcohols
Detergents Quats (cationic detergent) Phenolics Halogens Organic acids and bases Heavy Metals
Aldehydes
Ethylene Oxide Oxidizing agents
Denature proteins
Denaturing proteins Denature proteins
Aqueous ethanol (60-95%) and isopropanol are used as disinfectants Effectively kill bacteria and fungi but not endospores nor nonenveloped viruses
Alcohols
Used to mechanically wipe microbes off skin before injections or blood drawing.
Quartenary Ammonium compounds Quats are cationic detergents that act by disrupting lipid bilayers Quats are bactericidal, fungicidal, viricidal (enveloped), and amoebicidal are most effective against Gram-positive bacteria but do not kill endospores are rapidly inactivated by organics including cotton and soap ZephiranBenzalkonium chloride CepacolCetylpyridinium chloride
PHENOLICS
Carbolic acid 2-5%
Powerful microbicidal, very corrosive General purpose disinfectant in hospital
Two halogens are regularly employed as antimicrobials: Iodine and Chloride Chlorine: Used as a disinfectant (10% bleach) Chlorine: Hypochlorous acid (HOCl) is a product, formed in water, that is the active form of the disinfectant Chlorine: Applied in treatment of drinking water, swimming pool, and sewage
Halogens
Iodine:
Tincture of iodine (alcohol solution) was one of first antiseptics used. Combines with amino acid tyrosine in proteins and denatures proteins. Stains skin and clothes, somewhat irritating. Iodophors: Compounds with iodine that are slow releasing, take several minutes to act. Used as skin antiseptic in surgery. Not effective against bacterial endospores.
Betadine Isodine
Heavy Metals:
Include copper, selenium, mercury, silver, and zinc. Oligodynamic action: Very tiny amounts are effective. A. Silver: 1% silver nitrate used to protect infants against gonorrheal eye infections until recently. B. Mercury Organic mercury compounds like merthiolate and mercurochrome are used to disinfect skin wounds. C. Copper Copper sulfate is used to kill algae in pools and fish tanks.
Oxidizing Agents
It is not a terribly effective disinfectant or anticeptic This is because bacteria and body tissues contain enzymes (catalase) that inactivate hydrogen peroxide However, the oxygen released upon inactivation can help oxygenate deep wounds and thus kill strict-anaerobe contaminants, e.g., Clostridium tetani
Aldehydes:
Include some of the most effective antimicrobials. Inactivate proteins by forming covalent crosslinks with several functional groups. A. Formaldehyde gas: Excellent disinfectant. Commonly used as formalin, a 37% aqueous solution. Formalin was used extensively to preserve biological specimens and inactivate viruses and bacteria in vaccines. Irritates mucous membranes, strong odor. Also used in mortuaries for embalming.
Glutaraldehyde: Less irritating and more effective than formaldehyde. One of the few chemical disinfectants that is a sterilizing agent. A 2% solution of glutaraldehyde (Cidex) is:
Bactericidal, tuberculocidal, and viricidal in 10 minutes. Sporicidal in 3 to 10 hours.
Commonly used to disinfect hospital instruments. Also used in mortuaries for embalming.
Glutaraldehyde is capable of effective sterilizationat room temperature, even against endospores, and even in the presence of organics, but achieving sterilization requries many hours of exposure
Glutaraldehyde
ETHYLENE OXIDE
Highly inflammable, mixed with inert gases CO2, N Especially for heart lung machines, respirators, sutures, syringes, dental equipments (heat sensitive)
DYES
Combine with nucleic acids Aniline dyes
Brilliant green, malachite green, crystal violet
Acridine dyes
Proflavine, acriflavine, euflavine, aminacrine
Physical disinfectant
Agent
Mechanisms of Action
Denatures proteins
Denatures proteins Denatures proteins Incineration of contaminants Oxidation & Denatures proteins
Comments
Kills vegetative bacterial cells and viruses Endospores survive
121C at 15 p.s.i. for 30 min kills everything Kills pathogens in food products Used for inoculating loop 170C for 2 hours; Used for glassware & instrument sterilization
Physical Antimicrobials
Separation of bacteria Used for heat sensitive liquids from liquid (HEPA: from air) Desiccation and low temperature Decreased chemical reaction rate Used for food & drug preservation; Does not necessarily kill so used for Long-term storage of bacterial cultures Bacteriostatic
Plasmolysis of contaminants
Moist Heat
Moist heat kills microbes by denaturing enzymes (coagulation of proteins) Boiling (at 100C, I.e., at sea level) kills many vegetative cells and viruses within 10 minutes Autoclaving: steam applied under pressure (121C for 15 min) is the most effective method of moist heat sterilizationthe steam must directly contact the material to be sterilized Pasteurization: destroys pathogens (Mycobacterium tuberculosis, Salmonella typhi, etc.) without altering the flavor of the fooddoes not sterilize (63C for 30 seconds) Higher temperature short time pasteurization applies higher heat for a much shorter time (72C for 15 seconds) An ultra-high-temperature, very short duration treatment (140C for 3 sec.) is used to sterilize dairy products
Autoclave: Chamber which is filled with hot steam under pressure. Preferred method of sterilization, unless material is damaged by heat, moisture, or high pressure.
Temperature of steam reaches 121oC at twice atmospheric pressure. Most effective when organisms contact steam directly or are contained in a small volume of liquid.
All organisms and endospores are killed within 15 minutes.
Flaming
Incineration
Dont over load the oven Allow free circulation of air between the material
Earthenware filter
RADIATION
2 types
Non ionising
Infra Red radiation ( rapid mass sterilization of syringes, etc) Ultra Violet radiation (enclosed areas)
Ionising Gamma, X ray, cathode ray (plastics, syringes, oil, metal foils) Sterilization controls
Dosimeter measures radiation dose Colored discs M radiodurans, B pumilus
UV
C G
A T
A T
T A
G C
C G
A T
DNA
Industrial application
Cooling tower (Legionella control) Pharmaceuticals (disinfection of blood components and derivatives)