MICROBIAL CONTROL

Microbial control involves physical and chemical agents that destroy or inhibit microorganisms and
potential pathogens and prevent their transmission.

STERILIZATION

 It refers to the removal or destruction of all forms of life, including bacterial spores

PHYSICAL METHODS

I. APPLICATION OF HEAT
 heat is the most commonly used method for the removal of microorganisms
A. Moist Heat Procedure
 destroys microorganism by coagulation of enzymes and structural proteins and
degradation of nucleic acids
1. Boiling
- Destroys vegetative bacteria (non-sporulating)
- Temperature and time of exposure: 100°C for 10-15 minutes
2. Autoclave
- It is a chamber which is filled with hot steam under pressure
- Is the fastest and simplest method of sterilization- all organisms (except prions)
and spores are killed within 15 minutes
- It is used to sterilize biohazardous trash and heat-stable objects
- Principle: steam under pressure
- Biological indicator: B. stearothermophilus
a. 121°C, 15psi for 15 minutes- media, liquids, utensils, glass pipettes and
instruments for assay
b. 132°C, 15psi for 30-60 minutes- for decontaminating medical waste
3. Fractional/Intermittent Sterilization (Tyndallization)
- Temperature and time of exposure: 100°C for 30 minutes (3 consecutive days)
4. Inspissation
- It is used to sterilize protein-rich medium such as Lowenstein Jensen medium
- Principle: thickening of media through evaporation
- Temperature and time of exposure: 70°-80°C for 2 hours (3 consecutive days)
5. Pasteurization
- It is used to sterilize milk, dairy products, and alcoholic beverages
- It eliminates foodborne pathogens and organisms responsible for food spoilage
- This method cannot eliminate bacterial endospores
a. Low Temperature Holding (LTH)/ Batch Method
 Treatment at this temperature reduces spoilage of food without
affecting its taste
 It destroys milk-borne pathogens
 Temperature and time of exposure: 63°C for 30 minutes
b. High Temperature Short Time (HTST)/Flash Pasteurization
  Temperature and time of exposure: 72°C for 15 seconds (quick
heating then immediate cooling)
c. Ultra High Temperature (UHT)
 Temperature and time of exposure: 140°C for 3 seconds (cooled very
quickly in a vacuum chamber)

B. Dry Heat Procedure

 It kills microorganism by denaturation of proteins, sterilization without water
 It is utilized for the sterilization of glassware, oil products and powder
 It is not applicable for heat- sensitive materials
 Biological indicator: B. subtilis
1. Flaming
- Direct heating
2. Oven
- It is used for glassware, oil, petroleum of powders
- Temperature and time of exposure: 160°C to 170°C for 1.5 to 2 hours
3. Incineration
- Is the most common method of treating infectious waste and infected laboratory
animal
- Principle: burning materials into ashes (300°C to 400°C)
- Temperature of hazardous material (870°C to 980°C)
4. Cremation
- Used to control the spread of communicable disease

TERMINOLOGIES:

1. THERMAL DEATH POINT (TDP)- it refers to the lowest temperature at which a suspension of
bacteria is killed in 10 minutes
2. THERMAL DEATH TIME (TDT)- it refers to the shortest period of time needed to kill a
suspension of bacteria at a prescribed temperature and under specific conditions

II. FILTRATION
 It is the method of choice for sterilization of antibiotic solutions, toxins, chemicals,
radioisotopes, vaccines and carbohydrates (heat-sensitive solution)
2 TYPES OF FILTERS:
A. DEPTH FILTERS- consist of fibrous or granular material
B. MEMBRANE FILTERS- porous membranes; used to sterilize pharmaceuticals,
ophthalmic solutions, culture media, antibiotics, and oil products
1. Liquid filtration
- Uses cellulose acetate/ cellulose nitrate membrane with a vacuum
2. Air filtration
- Uses high- efficiency particulate air (HEPA) filters
- HEPA filters remove organisms larger than 0.3µm from isolation rooms, operating
rooms and biological safety cabinets
 3. Filtration of bacteria, yeasts, and molds
- Uses 0.45µm pores of membrane filters
4. Critical- sterilizing
- Uses 0.22µm membrane filters for parenteral solutions
III. LOW/COLD TEMPERATURE
 It is considered bacteriostatic because it reduces the rate of metabolism
 Important in food microbiology
IV. DESSICATION AND LYOPHILIZATION
 Destroys bacteria by disruption of metabolism; involves removing water from
microbes (bacteriostatic)
 Lyophilization is the most effective method for long term preservation of microbial
cultures
V. OSMOTIC PRESSURE
 Use of high concentrations of salts and sugars in food create a hypertonic
environment
VI. RADIATION
 When it passes through the cells, it creates free hydrogen and hydroxyl radicals and
some peroxidases which in turn cause different intracellular damage
1. Ionizing radiation
 Use: plastic syringes, sutures, catheters, gloves
2. Non-ionizing radiation
 Use: exposed surfaces, air, operating room
 Example: ultraviolet rays

CHEMICAL METHODS

DISINFECTION

 Refers to removal, inhibition or killing microorganisms including potential pathogens by using

chemical agents usually on inanimate objects; it does not remove all bacterial spores

TERMINOLOGIES

1. ANTISEPTIC- applied topically to skin; inhibits sepsis formation

2. DISINFECTANT- usually applied to inanimate objects
3. BACTERICIDAL- precipitates bacterial protein; it kills all bacteria in the specimen
4. BACTERIOSTATIC- inhibits the growth of organism

COMMON CHEMICAL AGENTS USED IN MICROBIAL CONTROL

1. Acid and Alkali solutions

o Hydrolyzes and coagulates proteins
2. Phenol and Phenolic compounds
o First widely used antiseptic and disinfectant
o Destroys plasma membranes and denatures cell proteins
 o Effective even in the presence of organics
3. Alcohol
o Causes denaturation of proteins and dissolution of lipid membrane
o Used both as an antiseptic and disinfectant
o Should be allowed to evaporate from the surface to which they were applied to achieve
maximum effectivity
4. Halogen
o Destroys microorganisms by oxidation mechanism
a. Iodophor
- Combination of iodine and neutral polymer (detergent)
b. Chlorine
- Used in the form of hypochlorite
- Should be made fresh daily
5. Salts of heavy metals
o Destroys microorganisms by inactivation and precipitation of cell proteins
6. Quaternary ammonium compounds
a. Detergent- widely used as surface- active agents
b. Phenolics- use: hospital floors and surroundings; found in germicidal soap
7. Aldehydes
o Used to sterilize medical instruments
a. Formaldehyde- usefulness is limited- has irritability factor and carcinogenic
b. Glutaraldehyde
8. Gas sterilant
a. Ethylene oxide
o Most commonly used gas for sterilization
o Use: plastic petri dishes, sutures
b. Peracetic acid