O2 + e -

O2 - Superoxide Superoxide
dismutase
O2 - + e - + 2H+ H2O2 Hydrogen
peroxide
H2O2 + e - + 2H+ Radical Catalase/
H2O + OH. hydroxil peroxidase
OH. + e - + 2H+ H2O water
 O2 and microbial growth
Classification Relationship with Explanation
O2
Aerobe
i. Obligate Necessary Presence of catalase
& SOD
ii. Facultative grow better with O2, Presence of catalase
anaerobes fermentative when lack of & SOD
O2
iii. Microaerophilic Needed but only small Absence of catalase &
amount of free O2 SOD. Pathogens uses
enzymes from the
host
Anaerobe
i. Aero tolerant Not needed, no difference Presence of SOD.
with O2. Undergo Other mechanisms is
anaerobic respiration present
ii. Obligate Sensitive towards O2 Absence of catalase &
Anaerobic
methods
 Water source example
activity aW
1.000 Pure water Caulobacter
0.995 Human blood Streptococcus,
Eschericia
0.980 Sea water Vibrio
0.950 Bread G+ve rod
0.900 Maple syrup G+ve coci
0.850 Smoked meat Yis (Sacharomyces
rouxii)
0.800 Jam, fruit cakes S. baili, penicillium
0.750 Salted fish Halobacterium
0.700 Dried fruits, cereals Xeromyces
bisporus
Generally microbe prefer aW range between 0.99-0.93
Fermentor/
bioreactor thermostat
probe
filter

Air (O2)

reservoir

pump temperature
(hotplate) Stirrer
(Mixing air
into medium)
 Microbial grouping according to their
energy & carbon source

Microorganism Energy Carbon examples

source source
Photoautotroph light CO2 Plants,
Photosynthetic
bacteria, algae
Photohetrotroph light Organic Purple & green non
compound sulphur bacteria
Chemoautotroph chemical CO2 Sulphur, iron
(inorganic) hydrogen bacteria
Chemoheterotroph chemical Organic Majority bacteria,
(organic) compound fungus, protozoa &
animals
Nutritional factors:

• Carbon Ingredient Quantity

• Nitrogen Glucose 25g
• Sulfur NH4Cl 3g
• fosforus KH2PO4 600mg
• traces K2HPO4 600mg
• water MgSO4 .7H2O 200mg
Growth factor
• Vitamins FeSO4 .7H2O 10mg
• Amino acids MnSO4 .4H2O 20mg
• Purine water Make up to 1 liter
Define medium
• pyrimidine
Adaptation to LIMITED nutrients
1. Synthesize increased amounts of enzymes for
uptake and metabolism of limited nutrients
2. Ability to synthesize enzymes needed to use a
different nutrient
3. Adjust the rate at which they metabolize nutrients
and rate at which they synthesize molecules
required for growth
No energy is wasted!!!
 SPORULATION

Bacteria Fungal
Endospore VS spore
-stationary phase
-great
-response to environment,
numbers
metabolic and cell cycle
-a form of
signals
reproduction
-form inside mother cell
when nutrient limited
-Not metabolically active
-Can survive long period
-Resistant to heat, radiation
& toxic

1 ENDOSPORE

1 PARENT
Sporulation SPORULATION

Vegetative sporulation
(METABOLICALLY ACTIVE)
 Endospore

• Resting stage during “lean or stressful times”.

• Develop in different locations of vegetative
cells:
•Schaeffer – Fulton Stain:
Young (24 h) Old (96 h)
Endospore
 STRESS
Spore
coat
keratin like-
proteins
resist toxic
chemicals.
Core Dipicolinic Acid & Cortex
Calcium protects DNA Protect core against osmotic
from heat. pressure-prevent drying
ENDOspore: sporulation
1) Core- contains
dipicolinic acid and Ca
ions- heat resistances-by
stabilizing protein
structure
2) Cortex- protects core
against changes- osmotic
pressure results from
drying
3) Spore coat- keratin
like protein resistance to
many chemicals
 Favorable Condition Return
Germination -
spore returns to its vegetative state
• 1)Activation - requires low pH/heat which damages the
coat
• 2)Germination - requires water and germination agent
(amino acid alanine/inorganic ions) that penetrates the
damaged coat.
• 3)Outgrowth - Proteins and RNA are synthesized.
• 4) Back to binary fission
 4) CULTURING BACTERIA

•Methods of obtaining pure cultures

•Culture media
•Methods of performing multiple diagnostic tests
 Streak plate: pure culture

Single colony-pure
culture

Methods of obtaining pure cultures

 Culture Media
Defined medium Complex medium

Contains reasonably familiar materials but varies

Contains known specific kinds & slightly in chemical composition from batch to batch
amount of chemical substances ie yeast, blood, soya bean, meat etc.
 Special medium for isolation & identification of
microorganisms

Medium Definition
Selective Encourages the growth of some
organisms BUT suppresses the growth of
others.
Differential Has a constituent that causes observable
changes in the medium (colour change).
Will facilitate to distinguish a certain type
of colony. Related to metabolism.
Enrichment Do not suppress others but contains
special nutrients that allow growth of a
particular organism
Urinary tract pathogens Fungal genus Candida

Differential media; CHROMagar

Maintaining cultures
1. Stock culture
2. Aseptic techniques
3. Preserved cultures
4. Reference culture – American Type Culture Collection ATCC
Methods of performing multiple diagnostic tests
Analytical Profile Index (API) 20E System