MICROBIOLOGY PHYSIOLOGY

NRG 107- Microbiology and Parasitology

LEARNING TARGETS:
1. PHYSIOLOGY. The study of how microbial
structures, growth and metabolism function
in living organisms.
2. NUTRITION. Substances that are acquired
from the environment and are used for
growth and metabolism.
3. GROWTH AND CONTROL. Involves IMPORTANT PROCESSESS
sterilization, disinfection, antisepsis,
sanitation, or degerming.  CARBOHYDRATE METABOLISM
o CELL RESPIRATION
 Aerobic
PHYSIOLOGY  Glycolysis
 Krebs Cycle
INTRODUCTION  Electron Transport Chain
(1 glucose mol. – 38 ATP)
Microbial Physiology has traditionally played a very
o FERMENTATION
important role in both fundamental research and in
industrial applications of microorganism. The  Anaerobic
classical approach in microbial physiology has  Glycolysis
been to analyze the role of individual components  End products are Lactic or
(genes or proteins) in the overall cell function. Ethanol
(1 glucose mol. – 2 ATP)
 LIPID METABOLISM
o Lipids are first broken down into
components fatty acids and glycerol
by lipases.
o Each component can then enter the
Krebs Cycle.
 PROTEIN METABOLISM
o Proteases and peptidases break
down protein into Amino Acids.
o Amino acids must undergo
enzymatic conversion into
substances that can enter Krebs
Cycle.
METABOLIC DIVERSITY

 PHOTOTROPHS- Light as energy source

 CHEMOTROPHS- Oxidation reduction of
inorganic and organic compounds
 AUTOTROPHS- self-feeders; capable of
producing its own food.
 HETEROTROPHS- Consumers; organisms
that eats other plants or animals for energy
and nutrients.
NOTE: most medical important organisms are the
CHEMOTOTROPHS, because typically, infectious
organisms catabolize substances obtained from the
host.
CHEMOTOTROPHS
 Energy and carbon both come from
inorganic compounds, and the same
compound can provide both. Specifically,
their energy source is electrons from
hydrogen atoms in organic compounds.
 Saprophytes- live on dead organic matter
 Parasites- nutrients from a living host
 This group (more precisely
chemoorganoheterotrophic) includes most
bacteria as well as all protozoa, fungi, and
animals.
 All microbes of medical importance are
included in this group.
MICROBIAL NUTRITION
I. TEMPERATURE
a. Psychrophiles- cold-loving
b. Mesophiles- moderate temperature
c. Thermophiles- heat-loving
Most bacteria grow within a limited range of
temperatures. Minimum and max growth
temperature 30 degrees celsius apart.
Optimum Temperature- are desired temperature at
which the species can best grow.
II. pH
 Most bacteria grow best between pH
range 6.5 to 7.5
 When bacteria are cultured in the
laboratory, they often produce acids
which interfere with their growth.
 Chemical buffers such phosphate salts
& peptones are included.
III. OSMOTIC PRESSURE
 Microbes obtain nutrients in solution
from water.
IV. ADAPTATIONS
a. Extreme halophiles- thrive in very
high salt concentrations
b. .Obligate halophiles- can only live in
high salt concentrations
c. Facultative halophiles- do not
require high salt concentrations but
can grow at concentrations up to
2%; both high and normal
concentrations.
CHEMICAL REQUIREMENTS
1. CARBON- Structural backbone of living
matter, needed for all organic compounds
that make up a living cell.
2. NITROGEN- For forming the amino group of
amino acids.
3. SULFUR- For synthesis of sulfur-containing
amino acid and vitamins such as Thiamine
(Vit. B1) and biotin (Vit. B7)
4. PHOSPHORUS- For the synthesis of
nucleic acids and phospholipids of the cell
membrane; also in ATP
5. TRACE ELEMENTS- Essential for some
enzymes, sometimes as cofactors; Includes
iron, copper, molybdenum and zinc
6. ORGANIC GROWTH FACTORS- Essential
compounds not synthesized which are
obtained from the environment; Include
enzymes for vitamin synthesis, amino acid,
purines, pyrimidines.
7. OXYGEN
a. Obligate aerobes- organism
requires oxygen to grow
b. Facultative aerobes- can use
anaerobic respiration or fermentation
 when oxygen is absent. (e.g. E. coli
and yeast)
c. Obligate anaerobes- cannot use
molecule oxygen for energy-yielding
reactions. (e.g. Clostridium)
d. Aerotolerant anaerobes- they can
survive and covert harmful forms of
oxygen to O2. (e.g. Lactobacilli)
e. Microaerophiles- can only tolerate
oxygen concentrations lower than
air. (e.g. Helicobacter pylori.)
TOXIC FORMS OF OXYGEN
The four forms of oxygen are SINGLE OXYGEN,
HYDROXYL RADICAL, PEROXIDE ANION, and
SUPEROXIDE RADICALS.
The single atom of oxygen (single oxygen) is highly
electronegative and causes toxicity in the cell. It is
associated with MUSCLE RIGIDITY,
CONVULSIONS, and SEIZURES as it reacts with
other biomolecules inside the cell.
SUPEROXIDE and HYDROXYL IONS are reactive
oxygen species and are produced when immune
cells are killing pathogenic cells. They are highly
toxic when in large concentrations and lead to the
formation of hydroxyl-anion, which further promotes
DNA DAMAGE.
PEROXIDE IONS damage the cell and result in
oxidative stress. They can react to different
chemical bonds, and atoms from the biomolecules
and disrupt the structure of the molecule, thereby
causing cell death.
GROWTH AND CONTROL
CULTURE MEDIA
 Nutrient material prepared for the growth
of microorganisms.
 Microbes that are introduced into a
culture medium to initiate growth are
called INOCULUM.
 Microbes that grow and multiply in a
culture medium is called a CULTURE.
 Culture media must be initially
“STERILE”.
USES OF CULTURE MEDIA
It is important to grow microorganisms outside the
body for the following purposes:
1. To identify the cause of infection from the
clinical sample, so that proper treatment can
be given.
2. To study the characteristics or properties of
microorganisms.
3. To prepare biological products like
vaccines, toxoids, antigens… etc.
TYPES OF CULTURE MEDIA
Classification based on PHYSICAL STATE:
a. Solid Medium- Agar is the most commonly
used solidifying agent.
b. Semi-Solid Medium- Such media are soft
and are useful in demonstrating bacterial
motility and separating motile from non-
motile strains.
c. Liquid Medium- Sometimes referred as
“broth; Bacteria grow uniformly producing
turbidity.
Classification based on INGREDIENTS:
a. Simple medium
b. Complex Medium
c. Synthetic Medium
d. Special Media
CULTURE MEDIA: AGAR
AGAR MEDIA is essential; for the study of
microorganisms and molecular biology and is
widely used in the culture and detection of
pathogens from contaminated food and water.
Due to its nature, agar is an aseptic and solidifying
component, which makes it an ideal candidate to
use in microbiology.
It liquifies at 100 degrees Celsius and remains
liquid until temp drops to 40 degrees Celsius.
NOT A NUTRIENT, but agar is mixed with nutrients
to create the perfect conditions for the development
of a microorganism.
 Used to grow bacteria that are exacting in
their nutritional needs. (e.g. Blood Agar and
Chocolate Agar)

b) SELECTIVE MEDIA- The inhibitory

substance is added to a solid media to
inhibit commensal or contaminating bacteria
such as: Antibitics, Dyes, Chemicals and
Alteration of pH
SELECTIVE MEDIA EXAMPLE: Thayer Martin
Medium selective for Neisseria gonorrhoeae
It usually contains the following combination of
antibiotics:

 Vancomycin: which is able to kill most

Gram-positive organisms.
 Colistin: which is added to kill most Gram-
negative organisms except Neisseria.
 Nystatin: which can kill most fungi
 Trimethoprim: which inhibits Gram-negative
organisms, specially swarming Proteus.

SPECIAL MEDIA
a) ENRICHED MEDIA- Substances like blood,
serum, egg are added to the simple media.
 d) TRANSPORT MEDIA
c) DIFFERENT MEDIA
 A media used for transporting the
 Are designed in such a way that
samples.
different bacteria can be recognized
 Delicate organisms may not survive
on the basis of their colony color.
the time taken for transporting the
 Dyes and metabolic substrates are
specimen without a transport media.
incorporated so that those bacteria
 Examples: Stuart’s Medium and
that utilize them appear as differently
colored colonies. Buffered Glycerol Saline
 Examples: MacConkey agar; CLED
agar; TCBS agar; XLD agar

SPECIAL CULTURE TECHNIQUES

 Mycobacterium leprae are grown in

armadillos.
 Treponema pallidum are grown in Rabbits.
 Bird egg and cultures parasites like
Chlamydias, Rickettsia’s and viruses.
PRESERVATION
a. REFRIGERATION for short term storage.
b. DEEP FREEXZING- pure culture of
microbes is placed in a suspending liquid
and quick frozen at temperaturesranging
from -50 degrees Celsius to -95 degrees
Celsius.
c. LYOPHILIZATION (Freeze Drying)-
suspension of microbes is quickly frozen -54
degrees Celsius to -72 degrees celsisu, and
the water is removed by a high pressure
vacuum.
FIVE “I”S CULTURING MEDIAS
1. INOCULATION- producing a pure culture.
2. ISOLATION- colony on media, one kind of
microbe, pure culture.
3. INCUBATION- growing microbes under
proper conditions.
4. INSPECTION- observation of
characteristics (data)
5. IDENTIFICATION- use data, correlation, to
identify organism to exact species.