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CONTROL OF MICROBIAL GROWTH JoyPCalayo, RMT,MSMT 2011 Terminologies 1. Sterilization: complete killing of all forms of microbial life 2.

Disinfection: process of reducing/eliminating pathogenic microorganisms in or on a material so that they no longer become a hazard; destruction of vegetative cells of pathogens 3. Decontamination: removal of foreign material/substances such as blood, serum and other biological fluids from items but does not necessarily mean removal of microorganisms 4. Antisepsis- destruction of vegetative pathogens present in living tissue 5. Degerming- mechanical removal of microbes from a limited area 6. Sanitization- employing of high temperature to lower microbial counts of materials to safe public health levels; treatment to reduce the number of disease-causing organisms to a level that is considered safe 7. Thermal death point (TDP)- lowest temperature at which all microorganisms in a particular liquid suspension will be killed in 10 minutes 8. Thermal death time (TDT)- minimum length of time for all microorganisms in a particular liquid suspension will be killed at a given temperature 9. Decimal Reduction Time (DRT, or D value)- time (mins) at which 90% of microbial population at a given temperature will be killed Methods of Microbial Control A. Physical B. Chemical A. PHYSICAL METHODS 1. Heat A. Dry heat a.1 Direct flaming Incineration a.2 Hot air sterilization or dry heat oven B. Moist heat b.1 Boiling b.2 Autoclaving: Routine/Typical Flash b.3 Pasteurization: Classic or LTH HTST UHT b.4 Tyndallization/fractional or intermittent sterilization b.5 Inspissation 2. 3. 4. 5. 6. 7. Dessication Filtration Radiation Use of high osmotic pressures Use of high pressures Use of low temperatures

Method of Microbial Control A. PHYSICAL 1. Heat a. Dry Direct flaming

Description/Mode of Action

OXIDATION OF CELL COMPONENTS Used for inoculating loops/needles by allowing them to turn red hot

Incineration

Used to destroy medical wastes & contaminated animal carcasses

Hot air/dry heat oven

Also denatures protein; for sterilizing laboratory glasswares, powders, oils and anhydrous materials; heat at 160-1700C for 2-3 hrs/ 160-1800C for 1.5-2 hrs

b. Moist

COAGULATION OF PROTEINS/PROTEIN DENATURATION Boiling Heating at 1000C for 5 mins/2-3 mins; cannot destroy endospores but can kill most microorganisms and viruses; can be used for drinking water Steam under pressure Heating at 1210C/15psi for 15-20 mins; will destroy endospores; used for microbiological media, medical/surgical items Heating at 1350C for 3 mins; to destroy prions, use 1320C for 4.5 hrs

Autoclaving

Pasteurization Batch : Classic/LTH

Used for milk, milk products, juices 60-630C for 30 mins

Flash: HTST UHT

720C for 15 secs 720C1400C720C for 5 secs Heat at 1000C for 30 mins and for 3 days; used to destroy vegetative forms of sporeforming bacteria Heat at 75-800C for 2 hrs for 3 days; used for high protein media Removal of water to avoid or disrupt microbial metabolism; e.g. freeze-drying or lyophilization Use of membrane filters (plastic polymers/ cellulose esters) or depth filters (fibrous/granules)to remove organisms from liquid or air; used for wines/beer, heatsensitive medications, BSCs or specialized labs and hospital rooms

Tyndallization/ fractional or intermittent sterilization Inspissation

2. Dessication

3. Filtration

4. Radiation Ionizing (gamma rays, X rays, high energy electron beams) Non-ionizing (UV and sunlight)

Destroys DNA; used for food preservation, medical/surgical supplies, medicines Damages DNA; used for vaccines, hospital rooms, operating rooms

5. Use of osmotic pressure

Causes plasmolysis; Use of high concentrations of sugars or salts; for food preservation Alters molecular structure of CHON and CHO at 130,000psi; for preserving fruit juices, colors, flavors and nutrient values DECREASES CHEMICAL REACTIONS & POSSIBLY CHANGES PROTEIN Bacteriostatic effect; for food,

6. Use of pressure

7. Use of Temp.

Refrigeration

drug, culture preservation Employed at -50 and -950C; for food, drug and culture preservation Done at -540C to -720C then dehydrated in vacuum; for food, drug and culture preservation

Deep-freezing

Lyophilization

B. CHEMICAL METHODS 1. Alcohols 2. Aldehydes 3. Biguanides 4. Gaseous sterilants 5. Halogens 6. Heavy metals and their compounds 7. Organic acids 8. Peroxygens 9. Phenols and phenolics 10. Surface-active agents MOA: 70% alcohol- lipid dissolution; protein denaturation Lysol: an ozone preparation of phenol; MOA: disruption of cell membrane, denaturation of proteins and enzymes Soap: disruption of cell membrane, saponification Chemicals Alcohols Characteristics Easy to obtain and inexpensive. Rapid evaporation limits their contact time Uses Aqueous solutions of alcohol are used as antiseptic to degerm skin in preparation for procedures that break intact skin, and as disinfectants fro treating instruments. Glutaraldehyde is widely used to sterilize medical instruments. Formalin is used in vaccine preparation Chlorhexidine is widely used as an antiseptic in soaps and lotions, and more recently, impregnated into catheters and surgical mesh Commonly used to sterilize medical devices Examples Ethanol, isopropanol

Aldehydes

Capable of destroying all forms of microbial life. Irritating to the respiratory tract, skin and eyes. Relatively low toxicity, destroys a wide range of microbes, adheres to and persists on skin and mucous membranes

Glutaraldehyde, formaldehyde

Biguanides

Chlorhexidine

Ethylene oxide gas

Easily penetrates hard to reach places and fabrics and does not damage moisture sensitive

Halogens

Metals

material. It is toxic, explosive and potentially carcinogenic Chlorine solutions are inexpensive and readily available; however, organic compounds and other impurities neutralize the activity. Some forms of chlorine may react with organic compounds to form toxic chlorinated products. Iodine is more expensive than chlorine and does not reliably kill endospores. Most metal compounds are too toxic to be used medically

Solutions of chlorine are widely used to disinfect inanimate objects, surfaces, drinking water and waste water. Tincture of iodine and iodophores can be used as disinfectants or antiseptics.

Chloride and iodine

Ozone

Peroxygen

Phenolic compounds

This unstable form of molecular oxygen readily breaks down Readily biodegradable and less toxic than traditional alternatives. The effectiveness of hydrogen peroxide as an antiseptic is limited because the enzyme catalase breaks it down. Peracetic acid is a more potent germicide than H2O2 Wide range of activity, reasonable cost, remains effective in the presence of detergents and organic contaminants, leaves an active antimicrobial residue.

Silver sulfadiazine is used in topical dressing to prevent infection of burns. Silver nitrate drops can be used to prevent eye infections caused by Neisseria gonorrhea in newborns. Used to disinfect drinking water and wastewater Hydrogen peroxide is used to sterilize containers for aseptically packaged juice and milk. Peracetic acid is widely used to disinfect and sterilize medical devices Triclosan is used in a variety of personal care products, including toothpastes, lotions, and deodorant soaps. Hexachlorophene is highly effective

Silver

Hydrogen peroxide and peracetic acid

Triclosan and hexachlorophene

Quaternary ammonium compounds

Non toxic enough to be used on food preparation surfaces. Inactivated by anionic soaps and detergents

against Staph aureus, but its use is limited because it can cause neurological damage Widely used to disinfect inanimate objects and to preserve non-food substance

Benzalkonium chloride and cetylpyridium chloride

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