MC3 – Microbiology & Parasitology
needed by microorganisms include:
JESSIE T. ORANO, RN, RM, MN, MAN, MMHA
DR. ARIES M. BALDONADO, RN, RM, LPT
WEEK 3 – PART I
MICROBIAL GROWTH
Learning Outcome:
1. Apply the principles of microbial growth in
understanding development of diseases.
MICROBIAL GROWTH
➢ Increase in number of cells, not cell size.
➢ One cell becomes colony of millions of
cells.
➢ Control of growth is
important for infection
control and growth of
industrial and biotech
organisms.
Microbial growth refers to an increase in
number of cells rather than an increase in
cell size. Many microbes (including
Escherichia coli and Salmonella enterica, are
unicellular, meaning they are made of only
one cell.
Bacteria divide and reproduce asexually.
Binary Fission and everything are equally
divided between the two daughter cells.
Time it takes for a single cell to go from one
division to the next: generation
time or doubling time. This is also the time it
takes for a population to double. For many
"typical" bacteria under "ideal" conditions this
doubling time may be as fast as 20 minutes.
Bacterial population
increases exponentially or logarithmically.
Factors Regulating Microbial Growth
• Nutrients
• Environmental conditions: temperature,
pH, osmotic pressure
• Generation time
Growth Factors - Microbes can exist in a great
many environments because they are small,
easily dispersed, need only small quantities of
nutrients, are diverse in their nutritional
requirements.
Nutritional (Biochemical) Factors – Nutrients
Carbon – carbon containing compounds are
needed as an energy source (ex. glucose) and for
building blocks.
Nitrogen - needed for amino acids and
nucleotides; some can synthesize all 20 amino
acids; others must have some provided in their
medium.
Sulfur – needed for amino acids, coenzymes,
Phosphorus – needed for ATP, phospholipids,
and nucleotides
Vitamins – a vitamin is an organic substance that
an organism requires in small amounts and that is
typically used as a coenzyme; many bacteria
make their own, but some are required in the
medium; microbes living in the human intestine
manufacture vitamin K, needed for blood clotting,
and some of the B vitamins, thus benefiting their
host.
Certain trace elements – ex. copper, iron, zinc,
sodium, chloride, potassium, calcium, etc.; often
serve as cofactors in enzymatic reactions.
CHEMICAL REQUIREMENTS
WATER!!!
ELEMENTS: CHONPS
TRACE ELEMENTS: Iron, Zinc,
Chromium, Selenium, Fluorine, Iodine
ORGANIC: Glucose, Vitamins, amino
Acids
NUTRITIONAL CATEGORIES
• SAPROBE – lives on organic matter of
dead organisms.
• PARASITE – lives on organic matter of
living host = pathogens.
ENVIRONMENTAL FACTORS
Temperature
O2
pH
Osmotic Pressure Others:
radiation, atmospheric pressure
 TEMPERATURE OPTIMA CATEGORIES Bacteria can be classified as:

Psychrophiles: cold-loving ✓ a. acidophiles (acid-loving) – grow

Mesophiles: moderate temperature-loving best at a pH of 1 to 5.4;
Thermophiles: heat-loving Ex. Lactobacillus (ferments milk)

Each has a minimum, optimum, and maximum ✓ b. neutrophiles – exist from pH to 5.4
growth temperature. to 8.5; most bacteria that cause human
disease are in this category.
Temperature – bacteria can be classified as:
✓ c. alkaliphiles (base loving) – exist
a. Psychrophiles (cold-loving) 15oC to 20oC; from pH to 7.0 to 11.5; ex. Vibrio
some can grow at 0oC. cholerae (causes cholera)
b. Mesophiles - grow best between 25oC and 40
C; human body temp is 37oC. BACTERIAL DIVISION
c. Thermophiles (heat-loving) – 50oC to 60oC;
BINARY FISSION
found in compost heaps and in boiling hot
springs. ✓ The process that bacteria use to carry out
cell division.
OXYGEN REQUIREMENTS
✓ Asexual reproduction by a separation of
Obligate aerobes – require O2.
the body into TWO new bodies.
Facultative anaerobes – can use O2 but also
grow without it. ✓ The organism duplicates its genetic
Obligate anaerobes – die in the presence of O2. material or the DNA and the divides into
two parts (CYTOKINESIS), with each new
organism receiving same copy of DNA.
Oxygen Requirements
1. strict or obligate anaerobes – oxygen kills
the bacteria; ex. Clostridium tetani
2. strict or obligate aerobes – lack of oxygen
kills the bacteria; ex. Pseudomonas
3. facultative anaerobes – can shift their
metabolism (anaerobic if oxygen is absent or
aerobic if oxygen is present); ex. E.
coli, Staphylococcus
4. aerotolerant – the bacteria don’t use
oxygen, but oxygen doesn’t harm them;
ex. Lactobacillus
5. microaerophiles – like low oxygen
concentrations and higher carbon dioxide
concentrations; ex. Campylobacter
pH CHARACTERISTICS
✓ Most bacteria grow between pH 6.5 and
7.5
✓ Acid (below pH 4) good preservative for
pickles, sauerkraut, cheeses
✓ Acidophiles can live at low pH
✓ Many bacteria and viruses survive low pH
of stomach to infect intestines.
✓ Helicobacter pylori lives in stomach
under mucus layer.
 OTHER ALTERNATIVES PROCESSES BY WHICH
MICROORGANISMS GROW AND
BUDDING:
MULTIPLY
✓ In this process a small bud-forms at one
end of the mother cell.
✓ As growth proceeds, the size of the
mother cell remains about constant, but
the bud enlarges. When the bud is about
the same size as the mother cell, it
separates.

GENERATION TIME
➢ Time required for cell to divide/for
population to double.
CONIDIOSPORES:
➢ Average for bacteria is 1-3 hours.
✓ Asexual reproduction among fungi.
E. coli generation time = 20 min. 20 generations
(7 hours), 1 cell becomes 1 million cells!

❖ LAG PHASE – making new enzymes in
FRAGMENTATION
➢ also known as splitting, is a form
of asexual reproduction in which an
organism splits into fragments.
❖ LOG PHASE – exponential growth.
➢ Each fragment develops into a mature
clone genetically and morphologically
identical to its parent.
PHASES OF GROWTH
response to new medium.
Lag Phase - In the lag phase, the number
of cells doesn't increase. However,
considerable metabolic activity is
occurring as the cells prepare to
grow. (This phase may not occur, if the
cells used to inoculate a new culture are in
the log phase & provided conditions are
the same).
✓ Desired for production of products.
✓ Most sensitive to drugs and
radiation during this period.
Log Phase (logarithmic or exponential
phase) - cell numbers increase
exponentially; during each generation time,
the number of cells in the population
increases by a factor of two). The number
of microbes in an exponentially increasing
population increases slowly at first, then
extremely rapidly. Organisms in a tube of
culture medium can maintain log growth for
only a limited time, as nutrients are used
up, metabolic wastes accumulate,
microbes suffer from oxygen depletion.
❖ STATIONARY PHASE – nutrients
becoming limiting or waste products
becoming toxic.
Stationary Phase - The number of cells
doesn't increase, but changes in cells
occur; cell become smaller and synthesize
components to help them survive longer
periods without growing (some may even
produce endospores); the signal to enter
this phase may have to do with
overcrowding (accumulation of metabolic
byproducts, depletion of nutrients, etc.).
death rate = division rate
❖ DEATH PHASE – death exceeds division
Death Phase - In this phase, cells begin to
die out. Death occurs exponentially, but at
a low rate. Death occurs because cell
have depleted intracellular ATP
reserves. Not all cells necessarily die
during this phase!
▪ DIRECT METHODS – count individual
cells.
▪ INDIRECT METHODS – Measure effects
of bacterial growth.
WEEK 3 – PART II
CONTROL OF MICROBIAL GROWTH
Learning Outcome:
1. Utilize techniques that effectively control
microbial growth.
CONTROL OF MICROBIAL GROWTH
❖ Control of microorganisms is 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. Physical agents include such
methods of control as high or low
temperature, desiccation, osmotic
pressure, radiation, and filtration.
❖ Control by chemical agents refers to the
use of disinfectants, antiseptics,
antibiotics, and chemotherapeutic
antimicrobial chemicals.
PHYSICAL AGENTS
❖ TEMPERATURE
➢ Temperatures below the
minimum usually have a static action on
microorganisms. They inhibit microbial
growth by slowing down metabolism but
do not necessarily kill the organism.
❖ TEMPERATURE
➢ Temperatures above the
maximum usually have a cidal action
since they denature microbial enzymes
and other proteins.
➢ Moist Heat: Autoclaving, Boiling Water
➢ Dry Heat: Hot air Sterilization, Incineration
➢ Pasteurization – mild heating of milk to kill
spoilage microorganisms or pathogens.
❖ DESICCATION
➢ Also known as drying, generally has
a static effect on microorganisms. Lack of
water inhibits the action of microbial
enzymes. Dehydrated and freeze-dried
foods, for example, do not require
refrigeration because the absence of
water inhibits microbial growth.
❖ OSMOTIC PRESSURE
➢ Movement of water through
semipermeable membrane from an area
of lower solute (greater water) to greater
solute (lower water) concentration.
➢ Water flows thru these processes:
Hypotonic (water flow into cell); Isotonic
(equal) and Hypertonic (water flows out of
cell).
❖ RADIATION
➢ Ultraviolet and Ionizing Radiation are
frequently used to reduce the microbial
populations in hospital operating rooms
and sinks, aseptic filling rooms of
pharmaceutical companies, in
microbiological hoods, and in the
processing, equipment used by the food
and dairy industries.
➢ Only microorganisms on the surface of a
material that are exposed directly to the
radiation are susceptible to destruction
and bacterial endospores are more
resistant to ultraviolet radiation. UV
radiation can also damage the eyes,
cause burns, and cause mutation in cells
of the skin.
❖ FILTRATION
➢ Microbiological membrane filters provide a
useful way of sterilizing materials such as
vaccines, antibiotic solutions, animal sera,
enzyme solutions, vitamin solutions, and
other solutions that may be damaged or
denatured by high temperatures or
chemical agents.
➢ The filters contain pores small enough to
prevent the passage of microbes but large
enough to allow the organism-free fluid to
pass through.
CHEMICAL AGENTS
❖ DISINFECTANTS, ANTISEPTICS, AND
SANITIZERS
➢ Disinfection is the elimination of
microorganisms, but not necessarily
endospores, from inanimate objects or
surfaces, whereas decontamination is
the treatment of an object or inanimate
surface to make it safe to handle.
CHEMICAL AGENTS
a. The term disinfectant is used for an
agent used to disinfect inanimate objects
or surfaces but is generally too toxic to
use on human tissues.
b. The term antiseptic refers to an agent
that kills or inhibits growth of microbes but
is safe to use on human tissue.
c. The term sanitizer describes an agent
that reduces, but may not eliminate,
microbial numbers to a safe level.
Factors influencing antimicrobial
action of disinfectants and antiseptics:
1. The concentration of the chemical
agent.
2. The temperature at which the agent is
being used. Generally, the lower the
temperature, the longer it takes to
disinfect or decontaminate.
3. The kinds of
microorganisms present. Endospore
producers such
as Bacillus species, Clostridium species,
and acid-fast bacteria like Mycobacterium
tuberculosis are harder to eliminate.
4. The number of microorganisms present.
The more microorganisms present, the
harder it is to disinfect or decontaminate.
5. The nature of the material bearing the
microorganisms. Organic material such
as dirt and excreta interfere with some
agents.
Modes of Actions of Disinfectants,
Antiseptics and Sanitizers:
➢ Damage the lipids and/or proteins
of the semipermeable cytoplasmic
membrane of microorganisms
resulting in leakage of cellular
materials needed to sustain life.
➢ They may denature microbial
enzymes and other proteins, usually
by disrupting the hydrogen and
disulfide bonds that give the protein its
three-dimensional functional shape.
This blocks metabolism.
Examples of Chemical agents commonly
used:
✓ Phenols and Phenolics – kills bacteria
by denaturing the protein cell membrane.
✓ Soaps and Detergents – mildly
antimicrobial; aids in mechanical removal
of microorganisms.
✓ Alcohol – 70% kills bacteria; once
evaporated, cidal effect ceases.
✓ Acids and Alkalis - alter membrane
permeability and denature proteins and
other molecules.
✓ Heavy Metals - such as mercury, silver,
and copper, denature proteins.
✓ Chlorine - reacts with water to
form hypochlorite ions, which in turn
denature microbial enzymes.
✓ Iodine and Iodophors – denature
proteins.
✓ Aldehydes - such as formaldehyde
(formalin) and glutaraldehyde, denature
microbial proteins.
✓ Peroxygenase - kills microorganisms by
oxidation and subsequent disruption of
their cytoplasmic membrane.
✓ Ethylene Oxide Gas – used for
sterilization (6-12 hrs. exposure).

Summary:
✓ Microbial growth refers to an increase
in number of cells rather than an
increase in cell size.
✓ Bacteria divide and reproduce
asexually.
✓ The control of microbial growth may
involve sterilization, disinfection,
antisepsis, sanitization.