Chapter 12.

Microbial Control
Principles and Terminology
• Main cellular targets of microbial control agents include proteins, DNA, and cell
membranes
o Protein Denaturation: deactivation of a protein through the loss of its high-
dimensional structure. Protein function depends on secondary, tertiary, and
quaternary structures. Protein folds are maintained by weak, low-energy
bonds such as hydrogen bonds, disulfide bonds, and ionic bonds. Many
agents, e.g., acids, oxidizing agents, heavy metals, cause protein
denaturation by disrupting these bonds.
o Surfactants: able to reach insoluble substance (e.g., grease, cell
membrane) through its long, hydrophobic hydrocarbon chains. Surfactants
(e.g., soap and detergents) controls microbial growth by disrupting cell
membranes.
• Sterilization: Destruction of all forms of microbial life (including endospores)
• Disinfection: Destruction of harmful microorganisms; do not achieve sterility.
(Chemicals used for disinfection are called "disinfectants")
• Antisepsis: Disinfection directed at living tissue. (Such a chemical is called an
"antiseptic")
• Bactericidal treatment:outright killing of bacteria (e.g., boiling water)
• Bacteriostatic treatment: inhibiting or stopping the growth of bacteria (e.g.,
refrigeration)

Physical methods of microbial control

• Dry heat (e.g. flaming the inoculation loop); mechanism: oxidation
• Pasteurization (low heat treatment of dairy products, e.g. 72oC/15 seconds; kills
most pathogens, but heat resistant bacteria can survive pasteurization)
• Autoclaving: use of high pressure moist heat; kept at 15 psi (121oC) for 20
minutes; preferred method of sterilization, because it kills all organisms including
heat-resistant endospores; moist is needed because protein denaturation occurs
most quickly in the presence of water. "Endospore strips" are included to ensure
the sterility of autoclaved materials.
• Filtration: for heat-sensitive chemical solutions (e.g., enzymes and vaccines), and
air
• Cold/Desiccation/Freeze-Drying(lyophilization): effects of low temperature and
desiccation (drying) are usually bacteriostatic and not bactericidal. Freezing and
freeze-drying are used for microbial preservation.
• Osmotic pressure: examples are the use of high concentrations of salt and sugar
for food preservation (causing "plasmolysis"of bacterial cells).
 • Radiation: ionizing radiation (e.g. gamma rays) and nonionizing radiation (e.g.,
UV light). They destroy microbes by damaging their DNA (DNA breakage by
gamma rays; thymine dimer formation by UV)
• Microwave radiation: causing the resonance of water molecules in food; do not
achieve sterility
• Sonication: use of sound wave for fragmenting cells to release its components.

Chemical methods
• Disk-diffusion method of evaluating a disinfectant: a disk of filter paper soaked
with a disinfectant can inhibit bacterial growth on a Petri dish, creating a "zone of
inhibition". The larger the zone, the more effective the agent is.
• Surface-active agents
o Soaps and detergents: Hand-washing remove microbes mechanically more
than through chemical killing. Soap dissolves oily film on skin surface, so
microbes can be lifted and removed through scrubbing
o Quats (quaternary ammonium)
o Mechanism: Surface-active agents dissolve cell membrane and cause
leakage of cell contents
• Acids and alkalies: e.g., organic acids; causing pH change, thus denaturing
proteins.
• Heavy metal compounds: use of organic copper, silver, and mercury compounds.
Cause protein denaturation by binding to sulfhydryl groups
• Halogens: e.g. chlorine used for drinking water and bleach for swimming pool
treatment; 1% alcohol solution of iodine is a common antiseptic. Halogens
denatures protein by oxidation (e.g., breaking disulfide bonds).
• Alcohols: the mechanism of action is protein denaturation by disrupting hydrogen
bonds; 70% alcohol is more effective than pure alcohol because the presence of
water makes protein denaturation quicker.
• Phenol and its derivatives: causing protein denaturation by disrupting hydrogen
bonds.
• Oxidizing agents: e.g., hydrogen peroxide.