MICROBIAL GROWTH

REQUIREMENTS;
Microorganisms must have the
proper PHYSICAL and
CHEMICAL conditions for
growth.
PHYSICAL REQUIREMENTS
I. Temperature
II. pH
III.Osmosis
I. Temperature; (3 groups of microorganisms
based on temp)

1. Psychorophiles; usually grow at temp between

0 and 20 degrees Celsius.

2. Mesophiles; usually grow at temperatures

between 20 and 40 degrees Celsius.

 Psychrotrophs are mesophilic organisms that

can grow in the psychrophilic range.

 These organisms are responsible for food

spoilage.

 Mesophiles include most of the common
spoilage and disease organisms. Ex. S.
aureus grows at 5 degrees C. in
refrigerated food and produce toxins.

 Proteus vulgairis causes eggs to spoil.

 Human pathogens are mesophiles

3. Thermophiles: capable of growth at high

temperatures, 40 to 90 degrees C.
 Extreme thermophiles have optimal growth at
80degrees or above.
 The known record for bacterial growth at high
temperatures is about 110 degrees C near deep-
ocean hydrothermal vents.
 Optimal growth temperature; temperature at
which the species grows best.
 Most mesophilic organisms have an optimal
growth temperature of approx. 37 degrees C.
 This means that most human pathogens are
mesophilic organisms.
II. PH: refers to the acidity or alkalinity of
a solution.'

 Most bacteria grow best at a pH range near

neutrality, between pH 6.5 and 7.5 ( neutrophilic).

 Few bacteria grow at a pH below 4.

 Acidophiles grow at the low pH and have been

found in acid having a pH of 1.

 Alkalinity also inhibits microbial growth but is

rarely used to preserve foods.

 Acid tolerant bacteria are valuable for the food and

dairy industries.
 Genera of Lactobacillus and
Streptococcus produce acid to convert
 milk to buttermilk,
 cream to sour
cream and milk curds to cheese.
 Acid in stomach deters disease in
organisms.
 Acid foods are rarely contaminated
III. Osmotic pressure;
 Osmosis; the movement of water from an
area of greater concentration of water to an
area of lower concentration of water
across a semi permeable membrane.

 Hypertonic solution; solution containing

more solutes and less water than the cells
suspended in it.
 Hypotonic solution; solution containing less
solutes and more water than the cells
suspended in it.
 Isotonic solution; solution containing the
same concentration of water as the cells
suspended in it.
 When microbial cells are in a hypertonic
solution, the water passes out of the cell
through the cell membrane, and the
membrane pulls away from the cell wall.

 This inhibits the growth of the

microorganism.
 Therefore the addition of salts (or other
solutes) to a solution, and the resulting
increase in osmotic pressure, can be used to
reserve foods .
 Salted fish, honey, and sweetened
condensed milk are preserved
largely by this mechanism: the high salt or
sugar cone.
 Draws water out of any microbial cells that
are present and thus prevent
their growth.
 Extreme halophiles; bacteria that have
adapted to high salt cone, and require them
for growth. Ex. Bacteria from dead sea
require nearly 30% salt.

Facultative halophiles: do not require high
salt cone. But are able to grow at salt cone,
up to 2%, a cone. That inhibits the growth
of many other bacteria.
 Most media used to grow bacteria is nearly
all water. Ex agar is usually only 1.5% salt.
CHEMICAL REQUIREMENTS;
 1. oxygen
 2. carbon
 3. nitrogen
 4. nutrients
1.Oxygen:
a. aerobic; organisms that use oxygen to produce
energy.

b. Obligate aerobes require oxygen to live.

c. Facultative anaerobes; use oxygen if present, but can
growing its absence.

d. Anaerobic; organism that do not use oxygen for

energy.

e. Obligate anaerobes; are unable to use molecular

oxygen for growth and are usually harmed by it. They
are unable to produce the enzyme superoxide
dismutases (SOD) to neutralize superoxide free
radicals
a. aerotolerant; organisms that cannot
use oxygen for growth but they
tolerate it fairly well. They produce
the enzyme superoxide dismutase.
b. Microaerophilic; organism that
require oxygen concentrations at
lower levels than those found in air.
c. Capnophilic; organisms that have
environment rich in carbon dioxide.
2. Carbon;
 Virtually all chemical substances in
microorganisms contain carbon in
some form.
 About 50% bacteria dry weight is
carbon.
 Both chemoautotrophic and
photoautotropic microorganisms
obtain their energy and produce their
nutrients from simple inorganic
compounds. Ex carbon dioxide
3. Nitrogen
 Used in synthesis of proteins, amino acids,
DNA, RNA.
 Bacteria that obtain nitrogen directly form
atmosphere are called nitrogen fixing
bacteria. Ex Rhizobium, Azotobacteria,
both found in soil.
4. Nutrients:
 Organisms need:
 a. water: solvent
 b. vitamins and minerals: coenzymes
 c. carbohydrates: energy source
 d. proteins: building blocks
 e. lipids: building blocks

 Heterotrophes: (other feeders) organisms that

obtain food from preformed organic carbon
compounds. Ex. All animals, most bacteria, and all
pathogenic bacteria.

 Autotrophes; (self feeder) organisms that synthesize

food from inorganic compounds.
Organisms synthesize food through 2
processes:
 Photosynthesis: organism takes carbon dioxide
from the air and water from the environment and
in the presence of energy from the sun converts
the carbon dioxide and water into glucose and
oxygen. EX green plants and preen and purple
sulfur bacteria.

 Chemosythesis: makes energy for ATP production

through chemical reactions taking place within
the organism.

Culture media
 a nutrient material prepared for the growth of
microorganisms in the laboratory.

 Criteria a culture media must meet.

 must contain the appropriate nutrients.

 must contain sufficient moisture.
 must contain a properly adjusted pH.
 must contain suitable levels of oxygen.
 media must initially be sterile.
 Culture media may be a liquid (broth) or a solid
(agar) . The agar is the solidifying agent and is
added to the nutrient broth.
 Complex media - (natural) not certain of exact
components of media. Media whose exact chemical
composition varies slightly from batch to batch. (Cheaper)
 Chemically Defined Media - nature and
quantity of each, component is known.
 The exact chemical composition is known.
Fastidious organisms require many growth
factors and do not grow well without
enriched media.
 Ex. Streptococcus - whole blood is added
to the agar.
Selective media
 Selective media - inhibits the growth of
unwanted bacteria, and encourages the
growth of desired bacteria.
 Ex EMB : Eosin Methylene Blue agar
contains carbohydrates fermented
by gram negative bacteria most contain
eosin and methylene blue that inhibits
gram positive organisms.
Differential media
 Differential media - media in which colonies of
different bacteria can be distinguished.
 Pure cultures of microorganisms have
identifiable reactions with differential media.
 Ex. a. Blood agar is used to identify bacterial
species that destroy red blood cells.
 Manitol salt agar - high salt concentration
inhibits most bacteria" and. The manitol may be
fermented to select for the growth
CULTURE MEDIA:
 Pure culture: if only one type of organism is present.

 Mixed culture : if 2 or more types of organisms are
present.
 Many media for microorganisms are complex reflecting the
growth requirements of the microorganism:

 Ex Rickettsia and Virus; must use living cells as

their media

 Anaerobic microorganism: atmosphere must be

free from oxygen ( placed w/in container where
CO2 and H2 are present)
 To preserve microbial cultures: place them in a refrigerator
which will slow down the metabolic reactions of the culture.
GROWTH OF BACTERIAL
CULTURES
 During their growth cycles,
microorganisms undergo reproduction
many times, causing the number in the
population to increase.
REASON FOR BACTERIAL
DEATH:
1. Unfavorable environmental
conditions.
2. Accumulation of waste.
3. Lack of nutrients.
Stages of bacteria growth ( in numbers)
1. lag phase - population remains temporarily
.unchanged at the beginning.. Lag phase usually
last for a few hours and would appear as a
straight horizontal line on a graph.
2. logarithmic (exponential) phase - Organism
..is expressing maximum reproductive potential.
There is a logarithmic (exponential) progression
of number is the population.
 During this time growth is the highest and
death is the lowest. at this time.
 Symptoms develop in humans and colonies
appear on the agar medium during the log
phase.
 This phase may last for hours or days.
3. stationary phase - living count remains
constant at the maximum value. Living count
levels off and would appear as a straight
horizontal line on a graph. The count remains
constant because
the same number of cells are dying as dividing.
This is due to the depletion of nutrients and the
accumulation of waste products. Spore-forming
bacteria will form spores during the stationary
phase.
4. Death phase - The living count decreases.
Death exceeds reproduction. Cells weaken,
change into odd shapes, undergo lysis and die.
The environment exerts LIMITING
FACTORS
 It has been calculated that if E. coli grew
unchecked for 36 hours, there would be enough
bacteria to cover the face of the earth.
 This can not happen due to limiting
environmental factors.
 In the lab a population of bacteria can be kept in
the log phase of growth indefinitely by using an
instrument called a chemostat.
 This instrument continuously drains off spend
media and adds fresh media.
Plate count method;

 Sample bacteria diluted, placed in petri

dishand set aside to incubate.
 Generally, plates with 30 to 300 colonies
are selected for determining final count,
which is expressed as number of bacteria
per original ml of sample.
 Most probable number; used to get
number of bacteria in contaminated water.
 ISOLATION OF
BACTERIA;
•
"To obtain separated colonies
from mixed culture
WAYS TO ISOLATE BACTERIA
1. Streak plate; sample of mixed bacteria is
streaked several times along one edge of petri
dish containing medium.

2. Pour plate method: a sample of bacteria is

diluted in several tubes of melted medium.
 After dilution, the melted agar is poured into
separate petri dishes and allowed to harden,
since bacteria have been diluted in various
tubes, the plates contain various dilution of
bacteria and where the bacteria are most
diluted, they form isolated
colonies.
3. Dry weight; liquid culture is dried out
and the amount of microbial
mass is weighed.
4. Turbidity method; assess bacteria
growth. As bacteria multiply in
liquid media, they make media cloudy.
 Placing culture tube in a beam of light and
noting the amount of light transmitted
gives idea of Turbidity of culture.
CONTROL OF MICROBIAL
GROWTH;
 Sterilization; destruction of all forms of
microbial life.
 Disaffection; destruction of pathogenic
organism on inanimate objects.
 Antisepsis; destruction of living object
such as skin surface.
 Sanitization; reduction in number of
pathogens to level deemed safe for public
health guidelines.
 Degerming; physical removal of
microorganisms by using soap ect...

Germicide; any chemical agent that kills
microorganism ( condition affecting
germicide include: 1. Type of
microorganism 2.Environment)
 Bactericide; kills bacterial
 Viricide: kill virus
PHYSICAL METHOD TO
CONTROL MICROBIAL GROWTH;
 Divided into : heat method and nonheat
method
 Thermal death point; lowest temp at
which microorganism are killed in 10 min,.
 Thermal death time; The minimal
amount of time needed to kill
microorganism.
 Decimal reduction; time for the
destruction of 90% of microbe population.
HEAT METHODS
 Dry heat; kill microorganisms by reacting with
oxidizing their protein. Ex Bunsen burner
 Moist heat; ex boiling water. Usually denature
microbe proteins ex autoclave
 Pasteurization; used to lower bacterial content
of milk and dairy products, it doesn't achieve
sterilization.
 It is set up to eliminate Tuberculosis bacillus and
rickettsia... pasteurize for 30 min at 62 C
for 15 to 27 sections at 72C
NON HEAT METHODS;
 filtration
 drying; when water is removed from cell
it shrivels and dies
 cold temperature; decrease microbial
metabolism.
 Radiation; gamma rays, x-rays
CHEMICAL METHOD:
 Most reduce microbial population to safe level or
remove pathogen from object.
 Among important criteria for selecting
antiseptic/disinfectant:
 Concentration of disinfectant
 whether agent is bactericidal or bacteriostatic
 nature of material to be treated (organic or not)
 temperature and pH that chemical will be used
at.
 time chemical agent will be left in contact with
surface
 Evaluation method; various dilutions of
chemical agent are prepared and tested
against equivalent dilution of phenol with
such bacteria as Staphylococcus aures
and Salmonella typhi
1. Phenol; one of first chemicals used for disinfecting (1860
Lister)
It damages cell membrane and inactivates enzymes of
microorganisms. Ex Lysol ( against Staphylococcus)
2. Halogens: ex Iodine ( betadine), cholorine
(bleach),ammonia
3. Alcohol: No effect on bacterial spores. Ex Isopropyl
(rubbing) alcohol evaporates quickly and leaves no
residue, degerms skin injections.
4. Heavy metals; ex silver nitrate: eyes of newborns to
guard against Neisseria gonorrhea and to cauterize
wounds, Copper sulfate: retard algae growth in pools,
Zinc Choride: mouth wash, Zinc oxide:
antifungal agent in paint
5. Soap and detergents; degerming
6. Aldehydes: ex glutaraldehyde: used to
sterilize hospital equip.
7. Oxidizing agents; Ex hydrogen
peroxide: kills microorganism by
releasing large amount of oxygen which
contributes to alteration of microbial
enzyme.
 ANTIBIOTICS;

The products of
microorganisms that react
with and inhibit growth of
other microorganisms.
1. Penicillin; prevents gram pos bacteria from
forming peptidoglycan
(main component of cell wall)
 If person is allergic the rxn may be
localized or whole body rxn ( anaphylaxis)
2. Cephalosporin; first produced by the mold
Cephalosprium and is useful against gram pos
bacteria and penicillin resistant bacteria.
3. Aminoglycoside; inhibit protein synthesis in
gram neg bacteria ex Streptomycin (fights
tuberculosis) many have debilitating side
effects.
4. Tetracycline; broad spectrum of drugs that inhibit gram neg
bacteria growth, rickettsia, chlamydia, some gram pos by
inhibiting protein synthesis.
 One side effect is to inhibit calcium deposit in body so shouldn't
be used in young children.

5. Other antibacterial antibiotics include;

a. Ervthromvcin; sub in for penicillin when allergy exists.

Useful against gram pos bacteria by inhibiting protein
synthesis.
b. Bacitracin; treat skin infection caused by gram pos
bacteria
c. Vancomycin; Effective against bacteria displaying
resistance to penicillin, cephalosporin and other
antibiotics, is very expensive and has may side effects, it
interferes with cell wall formation in b
d. Chloroamphericoil: against bacteria,
rickettsia, chlamydia.
 Has serious side effects such as aplastic anemia
(blood cells without hemoglobin), gray
syndrome (cardiovascular collapse)in babies, is
used for most serious bacterial infections ex
typhoid fever and meningitis
e. sulfa drugs: interfere with folic acid production
by bacteria. Ex sufamethoxazole, sulfisoxazole
against gram pos bacteria.
6. Antifungal drugs: ex griseofulvin ( ringworms,
athletes foot)
7. Antiviral drugs; not widely used, virus have
few functions or
structures with which drug can interfere. Ex
Azidothymidine (AZT), Acyclovir (herpes),
interferon (cancer)
8. Antiprotozoal drugs: many of the same sued
against bacteria work here ex tetracycline flagyl
( against trichomonas vaginalis), quinine
(malaria)
 COMMON PRESCRIPTION ABBREVIATIONS
 Ever wonder what those abbreviations on
your prescription mean?
 They tell the pharmacist how often or when
during the day you need to take a drug.
Here are a few of the most common ones:
 bid—twice a day qd—once a day
 hs—at bedtime qid—four times a day
 pe—after meals fid—three times a day
prn—as needed