ISOLATION OF PURE CULTURE OF ORGANISM & INHIBITION AND DESTRUCTION OF MICROORGANISMS

Kenneth S. Destura, RMT

Isolation of Pure Culture of Organism

ISOLATION OF PURE CULTURE

COLONIES

SERIAL DILUTION POUR PLATE

SERIAL DILUTION POUR PLATE

Nutrient Agar

SERIAL DILUTION POUR PLATE

SERIAL DILUTION POUR PLATE

Quebec Colony Counter

SERIAL DILUTION POUR PLATE

Formula:
Number of bacteria/mL = colony count x reciprocal of dilution
Number of colonies/plate = Ave. no. of colonies in 5 squares x 62.5

STREAK PLATE METHOD

STREAK PLATE METHOD

STREAK PLATE METHOD

STREAK PLATE METHOD

Inhibition & Destruction of Microorganisms

Inhibition and Destruction of Microorganisms

Definitions
Sterilization process whereby all forms of microbial life, including bacterial spores are killed Disinfection process whereby pathogenic organisms but not necessarily all microorganisms or spores are destroyed Antiseptic disinfectants used on living tissues Biocides also known as chemical sterilants, these are chemicals used to destroy all life Disinfectants chemical sterilants used for shorter periods

Inhibition and Destruction of Microorganisms

Definitions
Thermal death point lowest temperature at which all organisms in a liquid suspension can be killed in 10 minutes Thermal death time minimal length of time in which all bacteria in a liquid suspension will be killed at a given temperature -cide - a suffix indicating that the agent will kill the kind of organism in question (e.g. viricide) -static - a suffix indicating that the agent will prevent the growth of the type of organism in question (e.g., bacteriostatic)

Inhibition and Destruction of Microorganisms

Factors influencing antimicrobial activity:
the concentration and kind of a chemical agent used; the intensity and nature of a physical agent used; the length of exposure to the agent; the temperature at which the agent is used; the number of microorganisms present; the organism itself; and the nature of the material bearing the microorganism.

Physical Agents
Heat
Moist heat
Autoclave Boiling Tyndallization Pasteurization Inspissation

Radiation
Ionizing
Non-ionizing

Filtration Freezing
Deep Freezing Lyophilization

Dry heat
Hot air Sterilization Incineration

Ultrasound vibrations Lasers

Chemical Agents
PHENOLICS -laboratory and hospital disinfectants; act by denaturing proteins. ALCOHOLS -widely used disinfectants and antiseptics; will not kill endospores; act by denaturing proteins and possibly by dissolving membrane lipids. HALOGENS -widely used antiseptics and disinfectants; iodine acts by oxidizing cell constituents and iodinating cell proteins; chlorine acts primarily by oxidizing cell constituents. HEAVY METALS -effective but usually toxic; act by combining with proteins and inactivating them

Chemical Agents
QUATERNARY AMMONIUM COMPOUNDS -cationic detergents used as disinfectants for food utensils and small instruments, and because of low toxicity, as antiseptics for skin; act by disrupting biological membranes and possibly by denaturing proteins. ALDEHYDES -reactive molecules that can be used as chemical sterilants; may irritate the skin; act by combining with proteins and inactivating them STERILIZING GASES (e.g., ethylene oxide, betapropio-lactone) can be used to sterilize heat-sensitive materials such as plastic petri dishes and disposable syringes; act by combining with proteins and inactivating them. Recently, vapor-phase hydrogen peroxide has been used to decontaminate biological safety cabinet