PHYSICAL FACTORS AFFECTING BACTERIAL GROWTH

LBYBI13 (Lecture)
BIOMED | PROF. FLORA QUERUBIN | SEM 1 2023

○ Polaromonas vacuolata
CONCEPTS ● Mesophile
○ 20-45 deg. Centigrade
○ Escherichia coli
TEMPERATURE ● Thermophile
○ 45-68 deg. Centigrade
HOW DOES TEMPERATURE AFFECT MICROBIAL GROWTH? ○ Geobacillus stearothermophilus
● Increase in temperature: increased enzymatic ● Hyperthermophile
activity ○ 68 deg. Centigrade and up
○ Denaturation of enzymes
● Low temperature: decrease in enzyme activity pH
○ Freezing temperature stops enzymatic ● Measure of the activity of hydrogen ion or
activity (causing water to expand and hydrogen concentration
formation of ice crystals) leading to cell ● Neutral: pH 7
rupture ● Basic: > pH 7
● Acidic: < pH 7
CARDINAL TEMPERATURE Neutrophiles Optimally at pH value in the range
circumneutral (5.5-7.9)
Acidophiles < pH 5
Alkaliphiles > pH 8

HOW PH AFFECT AN ORGANISM?

● Extreme pH: affect structure of macromolecules
○ H bonds that holds together will break at
a high pH
○ Lipids are also hydrolyzed by basic pH
○ Proton motive force responsible for ATP
production is dependent on the
concentration gradient of hydrogen
across the plasma membrane.
● Moderate changes in pH would modify the
ionization of amino acid functional groups and
would disrupt hydrogen bonding, promoting
change in the folding of the molecule and
promotes denaturation and destroying the
● Minimum growth temperature: lowest activity.
temperature at which growth will occur
● Maximum growth temperature: highest
OSMOTIC PRESSURE
temperature in which growth can tolerate or
grow
● Optimum growth temperature: correlated with WHAT IS OSMOTIC PRESSURE?
the temperature of the normal habitat of the ● Pressure which needs to be applied to a solution
microorganisms to prevent the inward flow of water across a
semipermeable membrane
MAJOR GROUPS OF BACTERIA ACDG. TO TEMPERATURE REQUIRE. ● Depends on water availability in a cell
● Psychrophiles
○ 0-15 deg. centigrade
PHYSICAL FACTORS AFFECTING BACTERIAL GROWTH
LBYBI13 (Lecture)
BIOMED | PROF. FLORA QUERUBIN | SEM 1 2023
MICROORGANISMS CLASSIFIED ACCORDING TO TYPE OF OSM. PRE.
Halophiles Have specific requirements for NaCl (at least
some NaCl for their growth)
Halotolerant Organisms can tolerate some reduction in
their water activity of their environment but
grow best in the absence of the added solute
Extreme Capable of growth in a very salty environment
Halophiles
Osmophiles Able to live in a high sugar environment
ANAEROBIOSIS
MICROBES DISPLAY GREAT DIVERSITY IN THEIR ABILITY TO USE
AND TOLERATE OXYGEN
Obligate Rely on aerobic respiration for ATP; uses
oxygen as terminal electron acceptor in the
Aerobes
ETC; Fig. A
Microaerophiles Require oxygen for growth but are damaged
by normal atmospheric levels of oxygen and
don’t have ways to neutralize toxic forms of
oxygen such as as superoxide and hydrogen
peroxide.; Fig. D
Aerotolerant Do not use oxygen but can tolerate heat
because they have a fermentative metabolism
by which they are not harmed by the presence
of this oxygen; FIg. E
Facultative Most adaptable because capable of growing in
anaerobes oxic and anoxic environment; capable of both
fermentation and aerobic respiration. In case
there is a presence of oxygen, they would
utilize it well; Fig. C
Die in the presence of oxygen because they
Obligate
do not have the enzymes such as superoxide
anaerobes
dismutase and catalase that would turn
oxygen into non-toxic form; metabolize
through fermentation; Fig. B
Capnophiles Microoganisms that require high
concentration of carbon dioxide in order for
them to grow.
HOW OXYGEN CAN BE MANIPULATED IN THE LABORATORY?
● Use of thioglycollate medium
○ Composition of Thioglycollate
medium
■ Pancreatic digest of casein
■ Dextrose
■ Yeast Extract
■ Sodium Chloride
■ Sodium thioglycollate (reduces
oxygen to water)
■ Thioglycolic Acid (reduces
oxygen to water)
■ L-cystine (reduces oxygen to
water)
■ Resazurin (redox indicator,
colorless or white in anaerobic
condition but pink with the
presence of oxygen)
■ Agar- 0.75g/L (retardation of
oxygen diffusion & maintain
stratification of organism
growing in different layers of
blood, prevents convection
currents from carrying
atmospheric oxygen throughout
the blood)
● Manipulation of stab on solid tube cultures
○ Utilize butt or butt-slant tubed media
○ Anaerobic organisms will be able to
grow in the lower part/portion of the butt
tube.
● Use of layering technique
○ Aside from utilizing butt tube, we utilize
melted petroleum jelly/paraffin/oil in
layering the surface of the medium in a
culture tube. Enables an environment
with the juice oxygen in the tube set-up.
● Use of reducing agents (e.g. pyrogallol and
potassium hydroxide)
○ Pyrogallol and KOH are reducing agents
○ Seal the tube with rubber stopper and
seal with paraffin, a form of layering
technique.
○ What is the reason why we have to
invert the tube during incubation?
● Use of anaerobic jar systems (e.g. GASPAK)
○ Placed inside a polycarbonate char
along with gas generators and indicator
slip which is healed completely.
○ In the presence of water, chemicals that
are present inside the sachet such as
sodium bicarbonate and sodium
borohydride will chemically produce
hydrogen and CO2 gas. H reacts with O
PHYSICAL FACTORS AFFECTING BACTERIAL GROWTH
LBYBI13 (Lecture)
BIOMED | PROF. FLORA QUERUBIN | SEM 1 2023

present inside the jar which later on

produces water, forming a condensation
inside the jar. Reaction is catalyzed with
the element Palladium which is
observed in the figure attached to the
underside lead of the jar. The CO2 will
replace the removed oxygen, thus
creating a completely anaerobic
environment.
○ The effectiveness of the anaerobic jar
system can be via:
■ Indicator strip impregnated
with methylene blue which
remain colorless in an anaerobic
condition but will eventually turn
blue upon exposure to oxygen.
Can also be impregnated with
resazurin which becomes
colorless in an anaerobic
condition but turns pink in the
presence of oxygen.
■ Biological indicators such as
Pseudomonas which is
combined with other inoculated
plates for anaerobic culture.
Absence of growth for this
organism (obligate aerobe)
would indicate that perfect
anaerobiosis has been
achieved.

DESICCATION
● State of extreme dryness or drying out of a living
organisms
● When desiccated, microorganisms cannot grow
and divide but can survive depending on their
features.