Basic Principles of Microbial Control, and Control by Heat

Chapter 11

A. Definition of Terms: Know/look up meaning of:

1.Sterilization 2. Disinfection 3. Sanitization 4. Sepsis5. Asepsis6. Antiseptic-

A. Definition of Terms (cont.)

A. Definition of terms (cont.)

7. Germicide - A substance that kills "germs". Bactericide - A substance that kills bacteria. Algicide - A substance that kills algae. Fungicide - A substance that kills fungi. Viricide - A substance that "kills" viruses.

8. Microbistat (Germistat) - A substance that inhibits but does not kill microbes. a. Bacteriostat - A substance that inhibits bacteria. b. Fungistat - A substance that inhibits fungi. c. Algistat - A substance that inhibits algae.

Cellular targets of antimicrobial agents:

Fig. 11.4: Modes of action affecting protein structure

1. Cell wall 2. Cell membrane 3. Cellular synthetic processes (= synthesis of DNA, RNA, protein, etc.) 4. Proteins

B. Physical Cleanliness

Fig. 11.2: Factors affecting rate at which microbes are killed by antimicrobial agents

**See p. 15A packet Dirt such as blood, pus, soil, or other extraneous matter can reduce the effectiveness of antimicrobials Dirt can also protect microbes from exposure to antimicrobial agents

C. Killing curve: shows exponential reduction in microbial numbers Only a fraction of microbial targets die in a certain time interval So antimicrobial agents have gradual effect Longer exposure time results in more lethal hits The more microbial targets, the longer time needed to hit all targets

C. Killing curve (cont.)

Actively dividing cells (as in log phase of growth) are usually the most vulnerable to killing with antimicrobial agents Sometimes, surviving cells in stationary phase of growth are more resistant to antimicrobial agents than are log-phase cells

C. Killing Curve (cont.)

D. Use of heat to control microbes

Antimicrobial agents tested under defined conditions to determine microbial death rates This information can permit prediction of exposure time needed (under defined conditions) to kill a certain number of microbes

**See p. 15A packet; & pp. 326-330 1. Moist heat is the most effective form of heat for microbial control -Denatures proteins, damages membranes, & causes other types of cell damage (Moisture transmits heat more effectively than dry conditions)

D.1. Types of Moist Heat

D. 1. Moist Heat (cont.)

a) Boiling water, is 100oC (212oF) at sea level b) Free-flowing live steam, is 100oC, (212oF) at sea level Both these conditions will kill vegetative cells, but wont reliably destroy endospores

b. (1) Tyndallization: Heat material to kill vegetative cells Incubate material to allow endospore germination Heat material to kill new vegetative cells (that germinated from endospores)

D. 1. Moist Heat (cont.)

Fig. 11.5: Steam sterilization with autoclave

c) Compressed steam: at greater than 0.0 psi (elevated pressure), temperatures above 100oC can be achieved Autoclave (standard conditions to achieve sterilization are 15 psi, 121oC, 15-20 min); comparable to pressure cooker Allows reliable killing of endospores in reasonable amount of time

D. 1. Moist Heat (cont.)

d) Pasteurization: uses temperature and time of exposure that will not harm product (beverage) quality, but will kill pathogens and reduce number of spoilage microbes

D. 2. Dry Heat ** See p. 15B lecture packet

Moist heat kills faster than dry heat Dry heat must be used in some situations to sterilize substances that cannot be sterilized with steam Less efficient heat transmission with dry heat: compare hands in boiling water vs. oven at same temperature Dry heat causes cell oxidation, dehydration, protein denaturation

D. 2. Dry heat (cont.)

D. 2. Dry Heat (cont.)

To achieve sterilization with dry heat, temperature of 160o C or higher must be applied for 2 hours (or longer) Higher temperature needed to denature proteins with dry heat than with moist heat

Examples of use of dry heat: a) Oven; for glassware; 160oC b) Flame; 1,870oC max; metallic materials sterilized by incineration Incinerators; medical waste

3. Factors Affecting Heat Sterilization

a. Temperature b. Time of exposure c. Kind of medium conditions 1) Presence or absence of moisture a) Dissolved materials (interfere with killing) 2) pH; Killing more effective at pH extremes

3. Factors affecting heat sterilization (cont.)

d. Kind of organisms; spore formers, other thermodurics vs. more susceptible types e. Age of organisms, i.e. growth stage f. Load of organisms (numbers): greater numbers require longer exposure and/or higher temperatures

 Forms of heat provide broadspectrum, non-selective control of microbes

Cold E. Cold
1. Low temperatures - decrease microbial activity, but do not kill reliably a. Refrigeration: 5oC (41oF) slows metabolism and can be used for culture preservation

Cold (continued) E. Cold (cont.)

b. Freezing: -20o C ( -4o F ) (1) Slow die-off of microbes due to ice crystal formation which disrupts structures (membranes) or alters protein structure