MICROBIAL NUTRITION,

GROWTH AND METABOLISM

 Nutritional requirements
• Nutrients are substances used in biosynthesis and energy release
• Produce new cellular components and do cellular work
• THE COMMON NUTRIENT REQUIREMENTS
• Major elements: carbon, oxygen, hydrogen, nitrogen, sulfur, phosphorus
– Components of carbohydrates, lipids, proteins, and nucleic acids
– Skeletons or backbones of all the organic molecules
– Often satisfied together
– Electrons are very important
• provide energy for use in cellular work
• needed to reduce molecules during biosynthesis
• Minor elements: potassium, calcium, magnesium, and iron
– Cofactors, heat resistance and stability of ribosomes etc.,
 Nutritional requirements
• Microelements or trace elements: manganese, zinc, cobalt, molybdenum,
nickel, and copper
• Supplements : Vitamins, antibiotics
• Cannot be synthesized by itself but required for growth
– Diatoms need silicic acid (H4SiO4) to construct their beautiful cell
walls of silica [(SiO2)n]
– Mycoplasma need cholesterol
– Growth response assay
Balanced mixture of nutrients
NUTRITIONAL TYPES OFMICROORGANISMS
• Heterotrophs
• Organisms that use reduced, preformed organic molecules as their carbon
source
• The more reduced the organic carbon source (more electron), the higher its
energy content Ex: Lipids > Carbohydrates but not CO2
• Autotrophs
• Organisms that use CO2 as their sole or principal source of carbon
• Additional energy sources required - light or inorganic or organic
molecules
• Phototrophs - light as their energy source
• Chemotrophs - oxidation of chemical compounds (either organic or
inorganic)
• Based on the source of electron
– Lithotrophs – inorganic substances
– Organotrophs - organic compounds
 UPTAKE OF NUTRIENTS BY THE CELL
• Passive transport
• Facilitated Diffusion
• Ion channels
– Carrier mediated
• Active transport
– Pumps
– Transporters
 CULTUREMEDIA
• Suitable culture medium is required for growing microorganisms in the
laboratory
• Solid or liquid preparation used to grow, transport, and store
microorganisms
• Sources of energy, sources of electron, carbon, nitrogen, phosphorus,
sulfur, and various minerals
• Definite composition is dependent on the microorganisms
– Knowledge of a microorganism’s natural habitat
 CULTUREMEDIA
• Classification:
1. The chemical constituents from which they are made
2. Their physical nature
3. Their function
 CULTUREMEDIA
• Agar is a sulfated polymer composed mainly of D-galactose, 3,6-
anhydro-L-galactose, and D-glucuronic acid
• Why agar?
– Melting temp
– Hardening properties
– Undigested
• Koch, Loeffler and Minora Tarazaemon
Growth
 Growth curve
• Lag Phase:
– No immediate increase in cell number occurs
– Synthesizing new components
– Varies considerably in length
• Log Phase:
– Maximal rate of growth
– Rate of growth is constant
• Stationary Phase:
– Total number of viable microorganisms remains constant
– Balance between cell division and cell death
• Death and decline phase:
– Nutrient deprivation and the buildup of toxic wastes
 Growth curve
• Generation time or doubling time
MEASUREMENT OFMICROBIAL GROWTH
• Population number or mass
• Measurement of Cell Numbers
– Direct counting
– Easy, inexpensive, and relatively quick;
– Gives information about the size and morphology of microorganisms
– Petroff-Hausser counting chambers, hemocytometers, Coulter counter
and Flow cytometer
– Membrane filter technique
– Spread plate and pour plate techniques
Coulter counter
 MEASUREMENT OFMICROBIAL GROWTH
• Measurement of cell mass
• Indirect methods
• Microbial dry weight
– Cells growing in liquid medium are collected by centrifugation,
washed, dried in an oven, and weighed
– Time-consuming and not very sensitive
• Spectrophotometry
– More rapid and sensitive
– The amount of scattering is directly proportional to the biomass of cells
present and indirectly related to cell number
 THE CONTINUOUS CULTURE
OFMICROORGANISMS
• Nutrient supplies are not renewed nor wastes removed – Batch Growth
• How to maintain them in long log phase?
• Microbial population can be maintained in the exponential growth phase
and at a constant biomass concentration for extended periods in a
continuous culture system.
• Two types
Chemostats
• Sterile medium is fed into the culture vessel at the same rate as the media
containing microorganisms is removed
• Limiting nutrient
• Growth rate - rate at which new medium is fed
• Final cell density - concentration of the limiting nutrient
• D = f/V
• D – rate of nutrient exchange (dilution rate)
• f – medium flow rate
• V – vessel volume
 THE CONTINUOUS CULTURE
OFMICROORGANISMS
Turbidostats
• Photocell that measures the absorbance or turbidity of the culture in the growth
vessel.
• Flow rate of media through the vessel is automatically regulated to maintain a
predetermined turbidity or cell density
• Difference:
– Varying D in turbidostat
– No limiting nutrients
Water activity and osmotic pressure are
inversely related
Oxygen – electron
transport chain

Pressure – Barotolerant
– more than 500 atm

Radiations