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Chapter 4 Microbial Growth Measuring Microbial Growth

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12 views21 pages

Chapter 4 Microbial Growth Measuring Microbial Growth

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sheikhaishaa.aa
Copyright
© © All Rights Reserved
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Chapter 4

Microbial Growth Measurement

General Microbiology
Course Code: BIOL 230
Growth in bacteria: increase in number or
population
Binary fission: most of bacteria, exponential
growth
Budding: few bacterial species
Generation time

• The time required for a cell to divide ( population to double)


• Varies among species and with environmental conditions ( temperature)
• Can be as short as 20 minutes ( E. coli) to > 24h (Mycobacterium
tuberculosis)
• Most bacteria have a generation time of 1 to 3 hours

• Populations grow fast


• Eg, consider a 20 minute generation time
• 1 cell becomes 1 million in 20 generations (6.7 hrs)
• 1 billion in 30 generations (10 hrs)
Bacterial populations increase in number rapidly

The number of bacteria doubles in each generation. The superscript indicates the generation:
25= 5 generations
Bacterial Growth Curve: Arithmetic vs.
Logarithmic Representation
• It is difficult to graph
population change of such
big arithmetic numbers (
solid line)
Growth curve • Population changes cannot
plotted using be shown in the early stages
logarithmic of growth
scale • Hence we use log scales to
graph bacteria growth

Growth curve
plotted using
arithmetic
scale
Phases of
bacterial
growth:
Bacterial Growth Curve

• Lag Phase
• Period of little or no cell division / period of initial adjustment
• The physiological adaptation of the cell to the culture conditions.
• Horizontal line of the graph
• Last for several hours or several days
• Cells are not dormant
• Undergo intense metabolic activity (synthesis of enzymes and various
molecules )
Bacterial Growth Curve
Log/ exponential phase
• characterized by a period of the
exponential growth
• Cells begin to divide and enter period
of growth or logarithmic increase
• Most active cellular reproduction
phase
• Cells are most active metabolically
and is prefrrerd for industrial
purposes During exponential growth, the number of cells increases in the
geometric progression 20 , 21 , 22 , 23 , 24 until, after n divisions,
the number of cells is 2n. If the initial cell number is X0, the
number of cells after n doublings is X0 2n
Bacterial Growth Curve

• The stationary phase


• state of no net growth ( population stabilized)
• Although there is no net growth in the stationary
phase, cells still grow and divide. Growth simply
balanced by an equal number of cells dying
(reproduction rate equal to the equivalent death
rate)
• Growth rates slow down : number of microbial death
balances the number of new cells.
• Exhaustion of nutrients, accumulation of waste
products , production of secondary metabolites,
change in pH
Bacterial Growth Curve

• Death / decline phase

The final phase of the growth curve

• which is characterized by a net loss

of culturable cells – number of

deaths exceed number of new cell

cells
Direct Measurements of Microbial Growth
Plate counts
• Grow microbial sample on agar plate
• Count resulting colonies

Advantages – only viable (live) cells counted


Disadvantage – takes time for colonies to form
• CFU- Colony forming units
• Only limited colonies
should grow ( 25-300
colonies)
• Serial dilutions ensure
this range
Plate count : Pour plate or spread plate

Pour plate: not suitable for heat sensitive


microbes ( damaged by melted agar)
Most Probable Number (MPN)

• Statistical estimation technique based on

principle : greater the number of bacteria

the more dilution is required to a point

where no bacteria are left to grow

• Used when bacteria cannot grow on solid

media

• But, numbers are an approximation ~95%


accurate
Additional Direct Measurements

method of choice for low coun


Direct Measurements of Microbial Growth
Direct microscopic count: Counting chambers (slides-
Hemocytometer/ Petroff-Hauser chambers) for
microscope
• Number of microbes counted in microscope
• No incubation time required
• Most accurate, but …
• Motile cells difficult to count
• Dead cells look like live cells
• Need high cell numbers to count accurately
Estimating Bacterial Numbers by Indirect Methods

• Turbidity
• Cloudiness, or density, of a liquid culture
• Detected using a spectrophotometer at specific wavelgth of light
• Higher cell number = increased cloudiness/ turbidity

• Fast, convenient and easy method of obtaining number


• Nondestructive sampling
• No additional incubation
• but …
• Require large cell numbers to measure turbidity accurately
• Cannot distinguish between live and dead cells , clumping can cause
inaccurate results
Estimating Bacterial Numbers by Indirect Methods
Estimating Bacterial Numbers by Indirect Methods

• Metabolic activity
• Assumes higher number of bacteria produces higher amount of
metabolic product Eg, measure CO2 build-up

• Can be used when cells can’t be cultured

• Dry Weight :measuring dry weight of the microbial cells


• Removal of microbes from growth medium, dried, and weighed
• Useful for filamentous bacteria, molds
• Cannot disntinguish between live and dead ones
• Requires high concentration.

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