Ex.

28 Chemical Control of Microorganisms

Purpose of Microbial Control

Prevent Disease

Prevent Spoilage

Prevent Contamination

How do these agents work?

Destroy Essential Structures Destroy Essential Process

Effect on the Microorganism Bacteriostatic

Inhibition of growth and reproduction

Bactericidal
Ability to kill microorganisms

Terminology, cont.
Sterilization
Destruction of all forms of microbial life including endospores

Methods of Control
Physical
Heat Moist heat is most effective (pasteurization and canning) Cold Slows bacterial growth Can be used for long term storage

Ultraviolet Radiation Formation of lethal thymine dimers

Methods of Control, cont.

Physical, cont.
Desiccation Removal of water controls microbial growth Filtration Dependent on pore size of filter (some can remove viruses)

Methods of Control, cont.

Chemical Agents
Phenol and Phenolics- surface cleaner Biguanides (Chlorhexidine)- used as skin disinfectant in surgeries Halogens- iodine is a common antiseptic, chlorine a common disinfectant Alcohols- common disinfectant Heavy Metals- silver nitrate for treating gonococcal eye infections Surface-Active Agents- soaps and detergents Organic Acids- sorbic acid and benzoic acid are common Aldehydes- glutaraldehyde used in disinfecting medical equipment Gases- ethylene oxide (for heat sensitive objects) Peroxygens- ozone and hydrogen peroxide

Terminology
Disinfectant
Destruction of vegetative pathogens on inanimate objects

Antiseptic
Chemical agent designed for topical application to the body surface

Sanitizing Agent
Disinfectant that accomplishes some degree of cleaning Lowers microbial count on eating/drinking utensils to a safe public health level

Factors Influencing Effectiveness of Chemical Agent

Factors Influencing Effectiveness of Chemical Agent

Target organism Number of microbes (concentration) Microbial characteristics Environmental influences Time of exposure

Test Efficacy of Chemical Control

Tube dilution technique
Expose a test bacterium to a serial dilution of a chemical agent Tube with highest dilution (lowest concentration) where there is no visible growth is the minimum inhibitory concentration (MIC)

Test Efficacy of Chemical Control, cont.

Agar diffusion technique
Test organism is swabbed over the whole surface of an agar plate Holes can be punched in agar with chemical added or impregnated paper discs with chemical agent(s) are added Zone of no-growth is called zone of inhibition Size of the zone is NOT proportional to strength of chemical

Todays Procedure
Agar disc diffusion
Thoroughly swab a TSA plate with assigned bacterium made to a 0.5 McFarland Divide plate into quadrants with marker Add 5 impregnated paper discs to the plate as described by the instructor Incubate at 37C for 24 hours

Todays Procedure, cont.

1 2 3 4 5 6 7 8 9 10

Chemical agent

Control

Grab 10 tubes containing 1.0 ml of TSB (label each tube) Make a serial dilution of your chemical (control tube does not receive any of your chemical) Add 0.1 ml of your assigned organism made to a 0.5 McFarland to each tube Incubate tubes at 37C for 24 hours