Control of Microbial Growth

Introduction
• Early civilizations practiced
salting, smoking, pickling,
drying, and exposure of food
and clothing to sunlight to
control microbial growth
• Use of spices in cooking was to
mask taste of spoiled food.
Some spices prevented
spoilage.
Introduction
• In mid 1800s Semmelweis and Lister helped developed
aseptic techniques to prevent contamination of surgical
wounds. Before then:
• Nosocomial infections caused death in 10% of surgeries.
• Up to 25% mothers delivering in hospital died due to infection.
Definition of Terms
• Sterilization : killing or removing all forms of microbial life (including
endospores) in a material or an object.
• Heating is the most used method of sterilization
• Commercial sterilization:
• heat treatment that kills endospores of Clostridium botulinum the causative
agent of botulism, in canned food.
• does not kill endospores of thermophiles, which are not pathogens and may
grow at temperatures above 45°C
Definition of Terms
• Disinfection:
• reducing the number of pathogenic microorganisms to the point where they
no longer cause disease.
• Usually involves the removal of vegetative or non-endospore forming
pathogens.
• Might make use of chemical, radiation, boiling water or steam.
• Antiseptic :
• A type of disinfection where treatment is directed to living organism, the
chemical used is called antiseptic.
Definition of Terms
Modification of disinfection and antiseptic:

• Degerming
• mechanical removal of most microbes in a limited area.
• Example: alcohol swab on the skin

• Sanitization:
• use of chemical agent on food-handling equipment to meet public health
standards and minimize chances of disease transmission.
• Example: hot soap and water.
Definition of Terms
• Names of treatment that cause outright death of a microbes have the
suffix –cide, meaning kill.

• Biocide or germicide – kills microorganisms

• Fungicide –kills fungi
• Virucide - inactivates viruses
• Bactericide _ kills bacteria (not endospore)
• Sporicide - kills bacterial endospore
Definition of Terms
Bacteriostatic Agent
• Agent that inhibit the growth and multiplication of bacteria; their
names have the suffix –stat or –stasis, meaning to stop or steady.
• But once agent is removed, growth might resume
• Example: Bacteriostasis
Definition of Terms
Sepsis :
• comes from Greek for decay or putrid.
• Indicates bacterial contamination
Asepsis:
• absence of significant contamination
Aseptic techniques:
• used to prevent contamination of surgical instruments, medical
personnel and the patient during surgery.
• Also used to prevent contamination in food industry.
Rate of Microbial Death
Microbial Death Rate: An Example
Time Death per Minute Number of
• When bacterial populations Survivor
are heated or treated with 0 0 1,000,000
antimicrobial chemicals, they 1 900,000 100,000
usually die at a constant rate.
2 90,000 10,000
3 9,000 1,000
4 900 100
5 90 10
6 9 1

Meaning, for each minute the treatment is

applied, 90% of the remaining population is
killed.
Rate of Microbial Death
Factors that influence the effectiveness of antimicrobial treatments:

1. Number of microbes : the more microbes present, the more time it

takes to eliminate population.

2. Type of Microbes: endospores are very difficult to destroy.

Vegetative pathogens vary widely in susceptibility to different
methods of microbial control.
Rate of Microbial Death
Factors that influence the effectiveness of antimicrobial treatments:

3. Environmental Influences: Presence of organic material (blood,

feces, saliva) tend to inhibit antimicrobial, pH etc.

4. Time of exposure: chemical antimicrobial and radiation treatments

are more effective at longer times. In heat treatments, longer exposure
compensate for lower temperatures.
 Methods of Microbial Control
• Physical method of control • Chemical method of control
• Heat • Phenols and Phenolics
• Filtration • Biguanides
• Low temperature/cold • Halogens
• High pressure • Alcohols
• Desiccation • Heavy Metal and Their Compounds
• Osmotic Pressure • Surface Active Agents
• Radiation • Chemical Food Preservatives
• Aldehydes
• Gaseous Chemostetilizers
• Peroxygens
Physical Method of Control
Physical Method of Control
HEAT
• Common food preservation
• Kills microorganisms by denaturing their enzymes and other proteins.
Heat resistance varies widely among microbes.
• Thermal Death Point (TDP) : Lowest temperature at which all of the microbes
in a liquid suspension will be killed in ten minutes.
• Thermal Death Time (TDT) : minimal length of time in which all bacteria will
be killed at a given temperature.
• Decimal Reduction Time (DRT) : time in minutes at which 90% of bacteria at a
given temperature will be killed. Used in canning industry.
Physical Method of Control
HEAT
• Moist Heat – kills microorganisms by coagulating their proteins. It is
more effective than dry heat.
Examples:
• Boiling: Kills vegetative forms of bacterial pathogens, almost all
viruses, and fungi and their spores within 10 minutes or less.
Endospores and some viruses are not destroyed this quickly. However
brief boiling will kill most pathogens
• Hepatitis virus: can survive up to 30 minutes of boiling
• Endospores : can survive up to 20 hours or more of boiling
Physical Method of Control
HEAT
• Moist Heat
Examples:
• Autoclave: chamber which is filled with hot steam under pressure. More
reliable because temperature is above boiling point. Preferred method of
sterilization, unless material is damaged by heat, moisture, or high
pressure.
• Temperature of steam reaches 121°C at twice atmospheric pressure.
• Most effective when organisms contact steam directly or are contained in a small
volume of liquid.
• All organisms and endospores are killed within 15 minutes.
• Require more time to reach center of solid or large volumes of liquid
Physical Method of Control
HEAT
• Moist Heat
Examples:
• Pasteurization : used to reduce microbes responsible for spoilage of
beer, milk, wine, juices, etc.
• Classic Method of Pasteurization : milk was exposed to 65°C for 30 minutes.
• High Temperature Short Time Pasteurization (HTST) : used today. Milk is
exposed to 72°C for 15 seconds.
• Ultra High Temperature Pasteurization (UHT) milk is treated at 140°C for 3
seconds and then cooled very quickly in a vacuum chamber
• Advantage : milk can be stored at room temperature for several months.
Physical Method of Control
HEAT
• Dry Heat : kills by oxidation effects.
• Direct flaming: used to sterilize inoculating loops and needles. Heat metal
until it has a red glow.
• Incineration : destruction of something by burning, effective way to sterilize
disposable items (paper cups, dressings) and biological waste.
• Hot Air Sterilization : place objects in an oven. Requires 2 hours at 170 °C for
sterilization. Dry heat is transfers heat less effectively to a cool body, than
moist heat,
Physical Method of Control
FILTRATION – removal of microbes by passage of a liquid or gas through a
screen like material with small pores. Used to sterilize heat sensitive
materials like vaccines, enzymes, antibodies and some culture media.

• High Efficiency Particulate Air Filters (HEPA) : used in operating rooms and
burn units to remove bacteria from air.
• Membrane Filters : uniform pore size. Used in industry and research.
Different sizes :
• 0.22 and 0.45um Pores: used to filter most bacteria. Don’t retain spirochetes,
mycoplasmas and viruses.
• 0.01 um Pores : Retain all viruses and some large proteins.
Physical Method of Control
LOW TEMPERATURE – effect depends on microbe and treatment
applied.

• Refrigeration: temperature from 0 - 7°C. bacteriostatic effect. Reduces

metabolic rate of most microbes so they cannot reproduce or produce
toxins.
• Freezing: temperature below 0 °C
• Flash Freezing: does not kill most microbes
• Slow Freezing: more harmful because ice crystals disrupt cell structure
• Over third of vegetative bacteria may survive 1 year.
• Most parasites are killed by a few days of freezing.
Physical Method of Control
HIGH PRESSURE– if pressure is high enough, the molecular structure of
proteins and carbohydrates are altered, resulting into rapid inactivation
of vegetative bacterial cell.

• Endospores are highly resistant to high pressure. They can only be killed when
high pressure is combined with other techniques.
• Example: high pressure + high temperature,
Physical Method of Control
DESICCATION– absence of water wherein microorganism cannot grow
or reproduce but can remain viable for years.
when water is made available to them, they can resume their growth and division.

• Freeze drying of food is an example of desiccation

• The ability of dried microbes to remain viable is very important to hospital
settings. Dust, clothing, bedding, and dressing might contain infectious
microbes in dried mucus, urine, pus, and feces.
Physical Method of Control
OSMOTIC PRESSURE
• High concentration of salts and sugars to preserve food is based on
the effect of osmotic pressure.
• High concentration of these substances create hypertonic
environment that causes water to leave the microbial cell.

• Osmotic pressure is used in preservation of food: concentrated salt solutions

are used to cure meats, and thick sugar solutions are used to preserved fruits.
Physical Method of Control
RADIATION – three types of radiation kill microbes

1. Ionizing Radiation
• Gamma rays, X rays, electron beams, or higher energy rays. Have short
wavelengths (less than 1 nanometer), therefore carries more energy
• Cause mutations in DNA and produce peroxides.
• Used to sterilize pharmaceuticals and disposable medical supplies. Food
industry is interested in using ionizing radiation.

Disadvantages: penetrates human tissues. May cause genetic mutations in

humans.
Physical Method of Control
RADIATION – three types of radiation kill microbes

2. Ultraviolet light (Nonionizing Radiation)

• Wavelength is longer than 1 nanometer. Damages DNA by producing
thymine dimers, which cause mutations.
• Used to disinfect operating rooms, nurseries, cafeterias.

Disadvantages: damages skin, eyes. Doesn’t penetrate paper, glass,

and cloth
Chemical Method of Control
Chemical Method of Control
PHENOLS AND PHENOLICS
• Phenol (carbolic acid) was first used by Lister as a disinfectant.
• Rarely used today because it is a skin irritant and has strong odor.
• Used in some throat sprays and lozenges
• Acts as local anesthetic.
• Destroy plasma membranes and denature proteins
• Advantages: stable, persist for long times after applied, and remain
active in the presence of organic compounds.
Chemical Method of Control
PHENOLS AND PHENOLICS
• Phenolics are chemical derivatives of phenol
• Contain molecule of phenol that has been chemically altered to reduce its
irritating effect or increase its antibacterial activity in combination with a soap
or detergent
• Cresols: derived from coal tar (Lysol).
• Bisphenols: effective against gram-positive staphylococci and streptococci. It
is an ingredient of a prescription lotion used for surgical control procedure.
But, excessive use in infants may cause neurological damage.
Chemical Method of Control
PHENOLS AND PHENOLICS
• Bisphenols
• effective against gram-positive staphylococci and streptococci.
• It is an ingredient of a prescription lotion used for surgical control procedure.
• But, excessive use in infants may cause neurological damage.
Chemical Method of Control
BIGUANIDES
• Chlorhexidine
• Used for microbial control on skin and mucous membrane.
• When combined with alcohol, it is also used for surgical hand scrubs and
preoperative skin preparation.
• However, contact with eyes can cause damage.
• Biocidal against vegetative bacteria and yeast
Chemical Method of Control
HALOGENS – effective alone or in compounds.
• Iodine
• One of the oldest and most effective antiseptics.
• Effective to all form of bacteria, various fungi and some viruses
• It impairs protein synthesis and alters cell membranes
• Tincture (alcohol solution) was one of first antiseptic used.
• Stains skin and clothes, somewhat irritating
• Iodophors : compounds with iodine that are slow releasing take several minutes to
act. Less irritating and doesn’t stain. Used as skin antiseptic in surgery.
• Not effective against bacterial endospores.
• Betadine
• Isodine
Chemical Method of Control
HALOGENS – effective alone or in compounds.
• Chlorine
• it is used to disinfect drinking water, pools, and sewage.
• Easily inactivated by organic materials.
• Sodium hypochlorite : is active ingredient of bleach (Clorox)
When a quality of drinking water is a question, household bleach can provide a rough
equivalent municipal chlorination. After two drops of bleach are added to liter of water (4 drops
if water is cloudy) and the mixture sat for 30 mins. The water is considered safe to drink. FOR
EMERGENCY CONDITONS!
• Chloramines : consist of chlorine and ammonia. Less effective as germicides.
Chemical Method of Control
ALCOHOLS
• Kills bacteria, fungi, but not endospores or naked viruses.
• Act by denaturing proteins and disrupting cell membranes.
• Evaporate, leaving no residue.
• Used to mechanically wipe microbes off skin before injections or blood
drawing.
• Not good for open wounds, because it cause proteins to coagulate.
• Ethanol: drinking alcohol. Optimum concentrations is 70%.
• Isopropanol: rubbing alcohol. Better disinfectant than ethanol. Also, cheaper and
less volatile.
Chemical Method of Control
HEAVY METALS
• Include copper, selenium, mercury, silver, and zinc.
• Oligodynamic action: very tiny amounts are effective.

A. Silver
• 1% silver nitrate used to protect infants against gonorrheal eye infections until recently.
B. Mercury
• Organic mercury compounds like Merthiolate and mercurochrome are used to disinfect skin
wounds.
C. Copper
• Copper sulfate is used to kill algae in pools and fish tanks.
Chemical Method of Control
HEAVY METALS
D. Selenium
• Kills fungi and their spores. Used for fungal infections.
• Also used in dandruff shampoos.
E. Zinc
• Zinc chloride is used in mouthwashes.
Chemical Method of Control
SURFACE-ACTIVE AGENTS

• Soaps and detergents

• Little value as an antiseptic, but have an important function in mechanical
removing of microbes through scrubbing
• Acid-anionic sanitizers
• Very important in the cleaning of dairy utensils and equipment
Chemical Method of Control
SURFACE-ACTIVE AGENTS
• Quaternary ammonium compounds (quats)
• Widely used surface active agents
• Effective against gram positive bacteria, less effective against gram-negative bacteria.
• Also destroy fungi, amoebas, and enveloped viruses.
• Unknown chemical mode of action, but probably affect plasma membrane
• Zephiran, Cepacol, usually found in our lab spray bottles and mouthwash
• Pseudomonas strains are resistant and can grow in presence of Quats are a big concern in
hospitals.
• Advantages: strong antimicrobial action, colorless, odorless, tasteless, stable, and nontoxic.
• Disadvantages: form foam. Organic matter interferes with effectiveness. Neutralized by
soaps and anionic detergents
Chemical Method of Control
ALDEHYDES
• Include some of the most effective antimicrobials.
• Inactive proteins by forming covalent crosslinks with several functional groups.

A. Formaldehyde gas:
• Excellent disinfectant
• Commonly used as formalin, a 37% aqueous solution.
• Formalin was used extensively to preserve biological specimens and inactivate viruses
and bacteria in vaccines.
• It can irritate mucous membranes and has strong odor.
• Also used in mortuaries for embalming.
Chemical Method of Control
ALDEHYDES
B. Glutaraldehyde:
• Less irritating and more effective than formaldehyde.
• One of the few chemical disinfectants that is a sterilizing agent.
• A 2% solution of glutaraldehyde
• Bactericidal, tuberculocidal, and viricidal in 10 minutes.
• Sporicidal in 3 to 10 hours.
• Commonly used to disinfect hospital instruments.
• Also used in mortuaries for embalming.
Chemical Method of Control
GASEOUS STERILIZERS
• Chemicals that sterilize in a chamber similar to an autoclave.
• Denature proteins, by replacing functional groups with alkyl groups.

A. Ethylene Oxide:
• Kills all microbes and endospores but requires exposure of 4 to 18 hours.
• Toxic and explosive in pure form.
• Highly penetrating.
• Most hospitals have ethylene oxide chambers to sterilize mattresses and large
equipment.
Chemical Method of Control
PEROXYGENS (OXIDIZING AGENTS)
• Oxidize cellular components of treated microbes.
• Disrupt membranes and proteins.

A. Ozone:
• Used along with chlorine to disinfect water.
• Helps neutralize unpleasant tastes and odors.
• More effective killing agent than chlorine, but less stable and more expensive.
• Highly reactive form of oxygen.
• Made by exposing oxygen to electricity or UV light.
Chemical Method of Control
PEROXYGENS (OXIDIZING AGENTS)
B. Hydrogen Peroxide:
• Used as an antiseptic.
• Not good for open wounds because quickly broken down by catalase present
in human cells.
• Effective in disinfection of inanimate objects.
• Sporicidal at higher temperatures.
• Used by food industry and to disinfect contact lenses
C. Benzoyl Peroxide:
• Used in acne medications
Chemical Method of Control
PEROXYGENS (OXIDIZING AGENTS)
D. Peracetic Acid:
• One of the most effective liquid sporicides available.
• Sterilant:
• Kills bacteria and fungi in less than 5 minutes
• Kills endospores and viruses within 30 minutes
• Used widely in disinfection of food and medical instruments because it does
not leave toxic residues.
Chemical Method of Control
CHEMICAL FOOD PRESERVATIVES
• Sulfur dioxide
• Disinfectant in wine making
• Sodium nitrate and sodium nitrite
• Added to meat products like bacon, ham, hot dogs, etc.
• Preserve red color of meat and prevent growth of botulism endospores
• Sorbic acid
• prevent mold from growing in acidic food
• Calcium propionate
• Effective fungistat used in bread, prevent the growth of surface molds
 Thank You
For Listening 

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