MICROBIOLOGY AND PARASITOLOGY
CHAPTER 3 | LECTURE
CHAPTER 3: MICROBIAL GROWTH AND CONTROL
OUTLINE
I Control of Microbial Growth
A Controlling Microbial Growth
B Control of Microorganisms
C Control of Growth
D Microbial Growth
E Bacterial Growth Rate
II Control of Microbial Growth: Definition
A Several Factors influence the Effectiveness of
Antimicrobial Treatment
B Physical Methods of Microbial Control
C Chemical Methods of Microbial Control
D Chemical Antimicrobial Agents Mode of Action
E Factors Affecting the Antimicrobial Activity
F Qualities of Good Disinfectant
CONTROL OF MICROBIAL GROWTH
● 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. In mid 1800s Semmelweis and Lister helped
developed aseptic techniques to prevent contamination
of surgical wounds.
o Before then: • Nosocomial infections caused
death in 10% of surgeries. • Up to 25% mothers
delivering in hospitals died due to infection.
● Control of Microbial Growth means to inhibit or prevent
growth of microorganisms. This control is affected in two
basic ways: (1) by killing microorganisms or (2) by
inhibiting the growth of microorganisms.
CONTROLLING MICROBIAL GROWTH
● Important in the medical field, pharmaceutical and
biotechnology industries, academic research, and food
industry. Chemical Agents that can eliminate or suppress
microbial life are separated in different groups based on
their use.
CONTROL OF MICROORGANISMS
● Essential in order to prevent the transmission of diseases
and infection, stop decomposition and spoilage, and
prevent unwanted microbial contamination.
Microorganisms are controlled by means of physical
agents and chemical agents.
CONTROL OF GROWTH
● Usually involves the use of physical or chemical agents
which either to kill or prevent the growth of
microorganisms. Agents which kill cells are called cidal
agents; agents which inhibit the growth of cells (without
killing them) are referred to as static agents.
BACTERIAL GROWTH
● Is the asexual reproduction, or cell division, of a
bacterium into two daughter cells, in a process called
binary fission. Providing no mutational event occurs, the
resulting daughter cells are genetically identical to the
original cell.
MICROBIAL GROWTH
● Increase in number if cells rather than increase in size
● Understanding the requirements for microbial growth
o
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o Allow us to determine how to control the growth of
microbes
o Specifically, of those microbes that cause disease
and food spoilage
BACTERIAL GROWTH RATE
WHAT IS THE GROWTH RATE OF BACTERIA?
● The Growth Rate of Bacteria formula is defined as the
rate of exponential growth of a bacterial culture which is
expressed as generation time, also the doubling time of
the bacterial population and is represented as G = T/n or
generation_time = Time/No.of generation.
LAG PHASE
● Phase when bacteria absorb nutrients, synthesize
enzyme and prepare for reproduction.
LOGARITHMIC GROWTH PHASE OR
EXPONENTIAL GROWTH PHASE
● Phase when bacteria multiply so rapidly that the
population number doubles with each generation time.
Generations Time – time between the formation of a new
bacterium and its division into two daughter cells. Growth rate
is greatest during the logarithmic phase.
STATIONARY PHASE
● Phase when the rate of division slows and the number of
bacteria dividing equals the number of dying. It is during
this phase that its culture maintain its greatest population
density.
DEATH PHASE OR DECLINE PHASE
● Microorganism die at a rapid because of overriding, the
toxic waste products increase and the nutrients and
oxygen supply decreases.
CONTROL OF MICROBIAL GROWTH: DEFINITIONS
STERILIZATION
● Killing or removing all forms of microbial life (including
endospores) in a material or an object. Heating is the
most commonly 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 45oC.
● The complete destruction of all living organisms including
cells, viable spore, and viruses.
Sterilization can be accomplished by:
o Heat
o Heat and steam under pressure
o Gas (ethylene acide)
o Chemicals (formaldehyde)
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o Radiation of the ultraviolet rays or gamma rays ENVIRONMENTAL INFLUENCES

● Disinfection: Reducing the number of pathogenic
microorganisms to the point where they no longer cause
diseases. Usually involves the removal of vegetative or
non-endospore forming pathogens.
● May use physical or chemical methods.
o Disinfectant: Applied to inanimate objects.
o Antiseptic: Applied to living tissue (antisepsis).
o Degerming: Mechanical removal of most microbes
in a limited area. Example: Alcohol swab on skin.
o Sanitization: Use of chemical agent on food-handling
equipment to meet public health standards and
minimize chances of disease transmission. E.g: Hot
soap & water.
● Presence of organic material (blood, feces, saliva) tends
to inhibit antimicrobials, pH etc.
TIME OF EXPOSURE
● Chemical antimicrobials and radiation treatments are
more effective at longer times. In heat treatments, longer
exposure compensates for lower temperatures.
PHYSICAL METHODS OF MICROBIAL CONTROL
HEAT
● Kills microorganisms by denaturing their enzymes and
other proteins. Heat resistance varies widely among
microbes.

DISINFECTION THERMAL DEATH POINT (TDP)

● is the destruction or removal of infections or harmful
microorganism from non-living objects by physical or
chemical means.
o Eg. Pasteurization – use of disinfectants and use of
antiseptic
● Note: Disinfection is not sterilization because not all
microbes are destroyed in disinfection.
● 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 in
a given temperature.

Disinfectants – chemicals used to disinfect inanimate DECIMAL REDUCTION TIME (DRT)

objects like bedside and OR equipment ● Time in minutes at which 90% of bacteria at a given
o e.g. Cydex, chlorox, phenol, carbolic acid, eresol, temperature will be killed. Used in the canning industry.
xylenols and orthophenylphenol.
MOIST HEAT
SEPSIS ● Kills microorganisms by coagulating their proteins. In
● Comes from Greek for decay or putrid. Indicates bacterial general moist heat is much more effective than dry heat.
contamination. Asepsis: Absence of significant ● Reliable sterilization with moist heat that requires
contamination. Aseptic techniques are used to prevent temperatures above that of boiling water.
contamination of surgical instruments, medical
personnel, and the patient during surgery. Aseptic BOILING
techniques are also used to prevent bacterial ● Heat to 100 oC or more at sea level. Kills vegetative
contamination in food industry. forms of bacterial pathogens, almost all viruses, and fungi
and their spores within 10 minutes or less. Endospores
Bacteriostatic Agent An agent that inhibits the and some viruses are not destroyed this quickly. However
growth of bacteria, but does not brief boiling will kill most pathogens.
necessarily kill them.
● Suffix stasis: To stop or HEPATITIS VIRUS
steady.
● Germicide: An agent that ● Can survive up to 30 minutes of boiling.
kills certain
microorganisms. ENDOSPORES
Bactericide An agent that kills bacteria. Most ● Can survive up to 20 hours of boiling.
do not kill endospores.
Vericide An agent that inactivates the PASTEURIZATION
virus. ● Developed by Louis Pasteur to prevent the spoilage of
Fungicide An agent that kills fungi. beverages. Used to reduce microbes responsible for
Sporocide An agent that kills bacterial spoilage of beer, milk, wine, juices, etc.
endospores of fungal spores. o Classic Method of Pasteurization: Milk was
exposed to 65oC for 30 minutes.
● Rate of microbial death when bacterial populations are o High Temperature Short Time Pasteurization
heated or treated antimicrobial chemicals, they usually (HTST):
die at a constant rate. o Ultra High Temperature Pasteurization (UHT): Milk is
treated at 140oC for 3 seconds and then cooled very
SEVERAL FACTORS THAT INFLUENCE THE quickly in a vacuum chamber.
EFFECTIVENESS OF ANTIMICROBIAL ▪ Advantage: Milk can be stored for several
TREATMENT months.
NUMBER OF MICROBES
● The more microbes present, the more it takes time to DRY HEAT
eliminate population. ● Kills oxidation effects.
o Direct Flaming: Used to sterilize inoculating loops
TYPES OF MICROBES and needles. Heat metal until it has a red glow.
o Incineration: Effective way to sterilize disposable
● Endospores are very difficult to destroy. Vegetative
items (paper cups, dressings) and biological waste.
pathogens vary widely in susceptibility to different
o Hot Air Sterilization: Place objects in an oven.
methods of microbial control.
Require 2 hours at 170oC for sterilization. Dry heat

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transfers heat less effectively to a cool body, than
moist heat.
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,
antibiotics, and some culture media.
o High Efficiency Particulate Air Filters (HEPA):
Used in operating rooms and burn units to remove
bacteria from air.
o Membrane filters: Uniform pore size. Used in
industry and research.
▪ Different sizes: 0.22 and 0.45 um Pores:
Used to filter most bacteria. Don’t retain
spirochetes, mycoplasmas and viruses.
▪ 0.01 um Pores: Retain all viruses and some
large proteins.
LOW TEMPERATURE
● Effect depends on microbe and treatment applied.
REFRIGERATION
● Temperatures from 0 to 7 oC. Bacteriostatic effect.
Reduces metabolic rate of most microbes so they cannot
reproduce or produce toxins. u Freezing: Temperatures
below 0oC.
FLASH FREEZING
● Does not kill most microbes.
o Slow Freezing More harmful because ice crystals
disrupt cell structure.
● Over a third of vegetative bacteria may survive 1 year.
● Most parasites are killed by a few days of freezing.
DESICCATION
● In the absence of water, microbes cannot grow or
reproduce, but some may remain viable for years. After
water becomes available they start growing again.
Susceptibility to desiccation varied widely:
o Neisseria gonorrhea: Only survives about one
hour.
o Mycobacterium tuberculosis: May survive several
months.
o Viruses are fairly resistant to desiccation.
o Clostridium spp. and Bacillus spp.: May survive
decades
OSMOTIC PRESSURE
● The use of high concentrations of salts and sugars in
foods is used to increase the osmotic pressure and create
a hypertonic environment.
● Plasmolysis: As water leaves the cell, plasma
membrane shrinks away from cell wall. Cell may not die,
but usually stops growing.
o Yeasts and molds: More resistant to high osmotic
pressures.
o Staphylococci spp. that live on skin are fairly
resistant to high osmotic pressure.
RADIATION
● Three types of radiation that kill microbes:
o Ionizing Radiation: Gamma rays, X rays, electron
beams, or higher energy rays. Have short
wavelengths (less than 1 nanometer). Dislodge
electrons from atoms and form ions. Cause
mutations in DNA and produce peroxides. Used to
sterilize pharmaceuticals and disposable medical
supplies. Food industry is interested in using ionizing
radiation.
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▪ Disadvantages: Penetrates human tissues.
May cause genetic mutations in humans.
o 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.
o Microwave Radiation: Wavelength ranges from 1
millimeter to 1 meter. Heat is absorbed by water
molecules. May kill vegetative cells in moist foods.
Bacterial endospores, which do not contain water,
are not damaged by microwave radiation. Solid
foods are unevenly penetrated by microwaves.
Trichinosis outbreaks have been associated with
pork cooked in microwaves.
CHEMICAL METHODS OF MICROBIAL CONTROL
Types of Disinfectants:
PHENOLS AND PHENOLICS
● Phenols and Phenolics: u 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.
o Phenolics are chemical derivatives of phenol
o Cresols: Derived from coal tar (Lysol).
o Biphenols (pHisoHex): Effective against gram-
positive staphylococci and streptococci. Used in
nurseries. Excessive use in infants may cause
neurological damage.
o Destroy plasma membranes and denature proteins.
o Advantages: Stable, persist for long times after
applied, and remain active in the presence of organic
compounds.
HALOGENS
● Effective alone or in compounds.
Iodine: Tincture of iodine (alcohol solution) was one of first
antiseptics used.
o Combines with amino acid tyrosine in proteins and
denatures proteins.
o Stains skin and clothes, somewhat irritating.
o Iodophors: Compounds with iodine that are slow
releasing , take several minutes to act. Used as skin
antiseptic in surgery. Not effective against bacterial
endospores. Eg. Betadine u Isodine
Chlorine: When mixed in water forms hypochlorous acid: Cl2
+ H2O ------> H+ + Cl- + HOCl Hypochlorous acid u Used to
disinfect drinking water, pools, and sewage.
o Chlorine is easily inactivated by organic materials.
o Sodium hypochlorite (NaOCl): Is active ingredient
of bleach.
o Chloramines: Consist of chlorine and ammonia.
Less effective as germicides.
Alcohols: Kill bacteria, fungi, but not endospores or naked
viruses.
o Act by denaturing proteins and disrupting cell
membranes.
o Evaporate, leaving no residue.
o Used to mechanically wipe microbes off skin before
injections or blood drawing.
o Not good for open wounds, because cause proteins
to coagulate.
o Ethanol: Drinking alcohol. Optimum concentration is
70%.
o Isopropanol: Rubbing alcohol. Better disinfectant
than ethanol. Also cheaper and less volatil
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Heavy Metals: Include copper, selenium, mercury, silver, Peroxygens (Oxidizing Agents):
and zinc. o Oxidize cellular components of treated microbes.
o Oligodynamic action: Very tiny amounts are o Disrupt membranes and proteins.
effective. o Ozone:
▪ Silver: 1% silver nitrate used to protect infants ▪ Used along with chlorine to disinfect water.
against gonorrheal eye infections until recently. ▪ Helps neutralize unpleasant tastes and odors.
▪ Mercury: Organic mercury compounds like ▪ More effective killing agent than chlorine, but
Merthiolate and mercurochrome are used to less stable and more expensive.
disinfect skin wounds. ▪ Highly reactive form of oxygen.
▪ Copper: Copper sulfate is used to kill algae in ▪ Made by exposing oxygen to electricity or UV
pools and fish tanks. light.
▪ Selenium: Kills fungi and their spores. Used for o Hydrogen Peroxide:
fungal infections. Also used in dandruff ▪ Used as an antiseptic.
shampoos. ▪ Not good for open wounds because quickly
▪ Zinc: Zinc chloride is used in mouthwashes. broken down by catalase present in human
Zinc oxide is used as antifungal agent in paints. cells.
▪ Effective in disinfection of inanimate objects.
Quaternary Ammonium Compounds (Quats): ▪ Sporicidal at higher temperatures.
o Widely used surface active agents. ▪ Used by food industry and to disinfect contact
o Cationic (positively charge) detergents. lenses.
o Effective against gram positive bacteria, less o Benzoyl Peroxide: Used in acne medications.
effective against gram-negative bacteria.
o Also destroy fungi, amoebas, and enveloped
viruses. CHEMICAL ANTIMICROBIAL AGENTS MODE OF
Zephiran, Cepacol, also found in our lab spray ACTION
bottles. Pseudomonas strains that are resistant ● Damage to cell wall or inhibition of cell wall synthesis
and can grow in presence of Quats are a big ● Alteration of the cytoplasmic membrane permeability
concern in hospitals. ● Alteration of the physical and chemical state of proteins
▪ Advantages: Strong antimicrobial action, and nucleic acid
colorless, odorless, tasteless, stable, and ● Inhibition of enzyme function
nontoxic. ● Inhibition of protein and nucleic acid synthesis
▪ Disadvantages: Form foam. Organic matter
interferes with effectiveness. Neutralized by FACTORS AFFECTING THE ANTIMICROBIAL
soaps and anionic detergents.
ACTIVITY
● Concentration of the agent
Aldehydes:
o Include some of the most effective antimicrobials. ● Temperature (increase Temp., increase reaction)
o Inactivate proteins by forming covalent crosslinks ● Contact time
with several functional groups. ● pH
o Formaldehyde gas: ● Nature of microorganism (species)
▪ Excellent disinfectant. ● Growth phase of microorganisms
▪ Commonly used as formalin, a 37% aqueous ● Presence of special structure (spore capsule)
solution. ● Number of microorganisms
▪ Formalin was used extensively to preserve ● Presence of extraneous materials (blood, pus, etc)
biological specimens and inactivate viruses and
bacteria in vaccines. QUALITIES OF GOOD DISINFECTANT
▪ Irritates mucous membranes, strong odor. ● High co-efficient of disinfection
▪ Also used in mortuaries for embalming. ● Stable
● Water soluble
Glutaraldehyde: ● Non-toxic, non-corrosive
o Less irritating and more effective than formaldehyde. ● Easy to use
o One of the few chemical disinfectants that is a ● Cheap
sterilizing agent.
o A 2% solution of glutaraldehyde (Cidex) is:
▪ Bactericidal, tuberculocidal, and viricidal in 10
minutes.
▪ Sporicidal in 3 to 10 hours.
o Commonly used to disinfect hospital instruments.
o Also used in mortuaries for embalming.

Gaseous Sterilizers:
o Chemicals that sterilize in a chamber similar to an
autoclave.
o Denature proteins, by replacing functional groups
with alkyl groups.
o 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.

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CHEMOTHERAPEUTIC AGENTS
● Chemical substances used in treating diseases
REFERENCES
ANTIBIOTICS
● Special type of chemotherapeutic agents used in treating Notes from the discussion by
infectious diseases
● Usually obtained from living microorganisms. Virgen Milagrosa University Foundation powerpoint
presentation:

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