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TERMINOLOGIES Sterilization-destruction or removal of all forms of microbial life (including endospores) *Sterilant-sterilizing agent Commercial Sterilization-sufficient heat treatment

nt to kill endospores of Clostridium botulinum in canned food. Disinfection- destruction of vegetative pathogens Antisepsis- destruction of vegetative pathogens living on tissue Degerming/degermation- removal of microbes from a limited area, such as the skin around an injection site. Sanitization- treatment intended to lower microbial counts on eating and drinking utensils to safe public health levels. *Sepsis- comes from Greek for decay or putrid. Indicates bacterial contamination. Asepsis- absence of significant contamination. Aseptic techniques-used to prevent contamination of surgical instruments, medical personnel, and the patient during surgery. *Pasteurization- uses heat to kill pathogens and reduce the number of food spoilage microorganisms in foods and beverages *Chemotherapy- chemical agents to kill or inhibit the growth of microorganisms within host tissue *Suffix: cide- meaning kill Biocide, or germicide-kills microorganisms usually with certain exceptions, such as endospores); ex: fungicide, virucide -includes-ethylene oxide, propylene oxide, and aldehydes -also used in preserving specimens in laboratories *Suffix: static or stasis- to stop or to steady Ex: Bacterostatic, fungistatic, and visrustatic agent Gerrmistatic agents- refrigeration, freezing, and some chemicals Microbial Death Rate Microbial death- permanent loss of a microorganisms ability to reproduce under normal environmental conditions Microbial death rate- technique for the evaluation of an antimicrobial agent *Factors influencing the effectiveness of antimicrobial treatments: Population size Population composition Concentration or intensity of an antimicrobial agent Duration f exposure Temperature Local environment

Physical methods of microbial control Heat- dry heat, moist heat and pasteurization Low heat- freezing, regrigeration and lyophilisation Filtration High pressure Dessication Osmotic pressure Radiation

1. Heat : kills microorganisms by denaturing their enzymes and other proteins. Thermal death Point (TDP) lowest temperature at which all microbes in a liquid suspension will be killed in ten minutes. Thermal Death Time (TDT) minimal length of time in which all bacteria will be killed at a given temperature Decimal Reduction Time (DRT) time in minutes at which 90% of bacteria at a given temperature will be killed. Used in canning industry. A. Moist Heat Sterilization -kills microorganisms by coagulating their proteins -boiling or flowing steam and autoclave a. Boiling- kills vegetative forms of bacterial pathogens, almost all viruses dies within 10min Hepatitis virus: can survive up to 30minutes of boiling Endospores: can survive up to 20 hours or more of boiling *used in : Dishes, basins, pitchers, various equipments b. Autoclave -very effective method of sterilization -121C, 15 psi (pounds per square inch) -vegetative cells and their endospores are killed in about 15 min. -used in: Microbiological media, solutions, linens, utensils, dressings, equipment, and other items that can withstand temperature and pressure B. Pasteurization -heat treatment for milk that kills all pathogens and most non pathogens -developed by Louis Pasteur to prevent the spoilage of beer, milk, wine, juices, etc. Classic Method of Pasteurization milk was exposed to 65oC for 30 minutes High Temperature Short time Pasteurization(HTST) used today. Milk is exposed to 72oC for 15 seconds. Ultra High Temperature Pasteurization (UHT) milk is treated at 140oC for 3 seconds and then cooled very quickly in a vacuum chamber *Advantage- milk can be stores at room temperature for several months. C.Dry Heat -kills by oxidation, effects a. Direct flaming burning contaminants to ashes use: inoculating loops b. Incineration burning to ashes

*use: paper cups, contaminated dressings. Animal carcasses, bags and wipes c. Hot-air sterilization- oxidation -very effective method of sterilization but requires temperature of 170oC for about 2 hr. *use: empty glassware instruments, needles and glass syringes 2. Low Temperatures -inhibit their growth and reproduction by the use of either freezing or refrigeration -important in food microbiology (food, drug, and culture preservation) a. refrigeration- temperatures from 0 to 7oC. *bacteriostatic effect- reduces metabolic rate of most microbes so they cannot reproduce or produce toxins b. Freezing- an effective method for preserving microbial cultures on which cultures are quick-frozen between -50 and -95oC c. Lyophilization most effective method for long-term preservation of microbial cultures, water removed by high vacuum at low temperature 3. Filtration -removes microbes by passage of a liquid or gas through a screen like material -most filters in use consist of cellulose acetate or nitrocellulose -useful for sterilizing liquids (enzymes, vaccines) that are destroyed by heat Types of filters: Depth Filters- solution containing microorganisms is sucked through this layer under vacuum, and microbial cells are removed by physical screening or entrapment and also by absorption to the surface of the filter material. Membrane Filters- uniform pore size. Used in industry and research. -different sizes 0.22 and 0.45um pores- used to filter most bacteria. Dont retain spirochetes, mycoplasmas and viruses 0.01 um pores- retain all viruses and some large protein *Air can also be sterilized by filtration. Examples are surgical masks and cotton plugs. Laminar flow biological safety cabinets, which employ high efficiency particulate air (HEPA) filters (a type of depth filter) to remove 99.97% of 0.3m particles. 4. High Pressure -If the pressure is high enough, it alters the molecular structures of proteins and carbohydrates resulting in the rapid inactivation of vegetative bacterial cells. -preservation of colors, flavours, nutrient values Use: Fruit juices -endospores are relatively resistant to high pressure 5. Dessication -absence of water -involves removing water from microbes -primarily bacteriostatic -USE: food preservation -susceptibility to desiccation varies widely: Neisseria gonorrhoea : only survive about one hour

Mycobacterium tuberculosis : may survive several months Viruses are fairly resistant to dessication Clostridium spp. And Bacillus spp.: may survive decades

6. Osmotic Pressure -high concentration of salts and sugars in foods is used to increase the osmotic pressure and create a hypertonic environment. *Plasmolysis: As water leaves the cell, plasma membrane shrinks away from cell wall. Cell may not die, but usually stops growing. Yeasts and molds: more resistant to high osmotic pressure Staphylococcus spp. That live on skin are fairly resistant to high osmotic pressures 7. Radiation TYPES: 1. IONIZING RADIATION -destruction of DNA -gamma rays (cobalt), X-rays, electron beams, or higher energy rays -have short wavelengths (less than 1 nanometer) -the principal effect of ionizing radiation is the ionization of water, which forms highly reactive hydroxyl radicals. Disadvantages: Penetrates human tissues. May cause genetic mutations in humans. USE: Sterilizing pharmaceuticals and medical and dental supplies. *biosafety cabinets- contain UV light 2. ULTRAVIOLET LIGHT (NONIONIZING RADIATION) -wavelength is longer than 1 nanometer. Damages DNA by producing thymine dimers, which cause mutation. DISADVANTAGE: Damages skin, eyes. Doesnt penetrate paper, glass and cloth. USE: Control of closed environment with UV (Germicidal ) lamp (operating rooms, nurseries, cafeterias) 3. MICROWAVE RADIATION -wavelength ranges from 1mm to 1m -heat is absorbed by water molecules -may kill vegetative cells in moist foods -bacterial endospores, which do not contain water, are not damaged by microwave radiation -solid foods are unevenly penetrated by microwaves CHEMICAL AGENT USED TO CONTROL MICROBIAL GROWTH Phenols and phenolics Biguanides (chlorhexidine) Halogens Alcohols Heavy metals and their compound Surface active agents Chemical food preservatives Aldehydes Chemical sterilization Peroxygenes and other forms of oxygen

1. PHENOLICS Mechanisms of action: Disruption of plasma membrane and denaturation of enzymes. a. Phenol (carbolic acid) was first used by Joseph Lister --seldom used as a disinfectant or antiseptic because of its irritation qualities and disagreeable odor b. Phenolics -derivative of phenol that are reactive even in the presence of organic materials USE: environmental surfaces , instruments, skin surfaces and mucous membranes *eg: Cresol contains O-phenyl-phenol -common content of lysol c. Bisphenols -hexachlorophene-ingredient of a prescription lotion (pHisoHex) -used for surgical and hospital microbial control procedures -triclosan-ingredient in antibacterial soaps and atleast one toothpaste *DISADVANTAGE: may cause neurological damage when used excessively especially in babies.

2. BIGUANIDES -MOA- Disruption of plasma membrane USE: Skin disinfection especially for surgical scrubs -bactericidal to gram-positives and gram-negatives Chlorhexidine- microbial control on skin and mucous membrane 3. HALOGENS a. Iodine inhibits protein function and is a strong oxidizing agent -impairs protein synthesis and alters cell membranes apparently by forming complexes with amino acids and unsaturated fatty acids -water treatment Forms of iodine: Tincture solution in an aqueous alcohol Iodophor combination of iodine and organic molecule -do not stain and are less irritating -betadine- contains providone b. Chlorine Compounds: 1. Hypochlorous acid (HOCL) forms when chlorine is added to water -strong oxidizing agent that prevents much of the cellular enzyme system from functioning. a. Cl2 + H2O H+ + Cl- + HOCl b.HOCL H+ + OCl-

2. Liquid form of compressed chlorine gas for disinfecting municipal drinking water; water in swimming pools and sewage 3. Calcium hypochlorite Ca(OCI)2- used to disinfect dairy equipment and restaurant eating utensils 4. Sodium hypochlorite (NaOCl) used as a household disinfectant and bleach (Chlorox) and as a disinfectant in dairies, food processing establishments and hemodialysis systems. 5. Chlorine dioxide solution surface disinfectant and does not leave residual tastes or odors

6. Chloramines combinations of chlorine and ammonia -used in water treatment and sanitize glassware and eating utensils and to treat dairy and food manufacturing equipment 4. a. ALCOHOL MOA : Protein denaturation and lipid dissolution Bactericidal and fungicidal; but not effective against endospores or non-envelopes viruses Commonly used alcohols Ethanol- opt. Concn: 70% -it aids in the denaturation of proteins b. Isopropanol (rubbing alcohol)- slightly superior to ethanol as an antiseptic and disinfectant, less volatile, less expensive, and are easily obtained than ethanol 5. HEAVY METALS AND THEIR COMPOUNDS MOA: Denaturation of enzymes and other essential proteins Oligodynamic action- very small amounts of heavy metals to exert antimicrobial activity a. Silver nitrate used to prevent gonorrhoeal neonatal ophthalma *silversulfadiazine used as a topical cream on burns *surfacine- contains water insoluble silver iodide b. Mercury * mercuric chloride use as disinfectants primarily bacteriostatic c. Copper *copper sulfate used chiefly to destroy green algae (algicide) that grow on reservoirs, stock ponds, swimming pools and fish tanks d. Zinc * zinc chloride- mouthwashes *zinc pyrithione antidandruff shampoos e. Selenium -kills fungi ad their spores. Used for fungal infections -also used in antidandruff shampoos