PHYSIOLOGY OF BACTERIA

GROWTH AND MULTIPLICATION

• Bacteria divide – binary fission

• Nuclear division precedes cell division
• Cell divides – constrictive/pinching
• Ingrowth – transverse septum
• Generation time
– Coliforms – 20 minutes
– Tubercle bacilli – 20 hours
– Lepra bacilli – 20 days
GROWTH AND MULTIPLICATION

• Multiplication - arrested – depletion of nutrients,

accumulation of toxic products
• Chemostat or Turbidostat - special devices
• Replenishing nutrients
• Maintain continuous cultures

• Industrial/research purpose
• Host tissue – nutrients may be inexhaustible but
parasite has to contend – host defence
GROWTH AND MULTIPLICATION

• Bacterial growth – two levels

– Increase in size
– Increase in number
• When critical size reached – cell divides
• Growth in number – bacterial count
• Total count
• Viable count
GROWTH AND MULTIPLICATION

• Total count – total number of cells whether living or

not
• Methods used
– Direct counting – counting chamber
– Electronic device – Coulter counter
• Direct counting – stained smear
• Relative numbers - in smear
• Opacity measure – nephelometer
• Separation of cells – centrifuge
• Chemical assays – Example: Nitrogen
GROWTH AND MULTIPLICATION

• Viable count – measures number of living cells

• Obtained by

– DILUTION methods

– PLATING methods
VIABLE COUNT – DILUTION METHOD

• Suspension diluted to point beyond which unit

quantities do not yield growth
• Tubes inoculated with varying dilutions
• Viable count calculated from the number of tubes
showing growth
• Application - bacteriology of water
• ‘Presumptive coliform count’
VIABLE COUNT - PLATING METHOD

• Dilutions inoculated on solid media

• Surface plates or pour plates
• Colonies that develop after incubation – viable count
• Method described by Miles and Misra (1938)
commonly used
BACTERIAL GROWTH CURVE

• Bacterium seeded – suitable liquid media

• Growth follows – definite course bacterial count –
made at intervals after inoculation
• Plotted in relation to time
• GROWTH CURVE
BACTERIAL GROWTH CURVE

Bacterial growth curve. The viable count shows the lag, log, stationary and
decline phases. In the total count, the phase of decline is not evident.
BACTERIAL GROWTH CURVE

LAG PHASE – adapt to new environment

• Enzymes and metabolic intermediates built
• Increase in size of cells
LOG PHASE – (logarithmic or exponential phase)
• Cell starts dividing
• Numbers increase exponentially or by geometric
progression
BACTERIAL GROWTH CURVE

STATIONARY PHASE
• After exponential growth – cell division stops due to
depletion of nutrients and accumulation of toxic
products
• Progeny cells replace cells that die
• Viable count - stationary
BACTERIAL GROWTH CURVE

PHASE OF DECLINE
• Population decreases due to cell death, nutritional
exhaustion
• Toxic accumulation
• Autolytic enzymes
• Total count – parallel to viable count up to stationary
phase and continues without phase of decline
MORPHOLOGICAL AND PHYSIOLOGICAL ALTERATIONS

• Lag phase – maximum cell size

• Log phase – cells smaller
• Stain uniformly
• Stationary phase – Gram variable
• Irregular staining
• Sporulation
• Metabolic products
• Phase of decline – involution forms
BACTERIAL NUTRITION

• Phototrophs - energy from sunlight

• Chemotrophs – energy from chemical reaction
• Autotrophs – synthesise all their organic compounds
• Heterotrophs – unable to synthesise their own
metabolites and depend on preformed organic
compounds
OXYGEN REQUIREMENTS

Aerobic bacteria
• Obligate aerobes – grow only in presence of oxygen
• Facultative aerobes – aerobes but can grow in
absence of oxygen
OXYGEN REQUIREMENTS

Anaerobic bacteria
• Grow in absence of oxygen
• Obligate anaerobes – may die on exposure to oxygen
• Microaerophilic - grow best in the presence of low
oxygen tension
OTHER REQUIREMENTS

• CARBON DIOXIDE - all bacteria require small

amounts of carbon dioxide for growth
• Capnophilic bacteria require higher levels – 5-10 %
for growth
• TEMPERATURE – optimum temperature most
pathogenic bacteria = 37°C
– Mesophilic bacteria – 25-40°C
– Psychrophilic bacteria - <20°C
– Thermophilic bacteria – 55-80°C
OTHER REQUIREMENTS
• Thermal death point – lowest temperature that kills
a bacterium under standard conditions in a given
time
MOISTURE and DRYING
• Water is essential – bacterial protoplasm
• Drying – lethal
• Spores – resistant
• Lyophilisation – freeze drying - drying in vaccum in
cold - preservation of bacteria, viruses
OTHER REQUIREMENTS

H-ION CONCENTRATION
• Bacteria sensitive to variations in pH
• Majority of pathogenic bacteria –neutral or alkaline
pH (7.2-7.6)
LIGHT
• Bacteria grow well in dark
• Sensitive to UV light
• Die on exposure to sunlight
• Photochromogens – produce pigment on exposure to
light
OTHER REQUIREMENTS
OSMOTIC EFFECT
• Tolerant to osmotic variation
• Plasmolysis - sudden exposure – hypertonic solution
– osmotic withdrawal of water
• Shrinkage of protoplasm
MECHANICAL AND SONIC STRESS
• Tough cell walls
• Ruptured – mechanical stress
• Disintegrate – ultrasonic vibration