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This document defines and describes various aspects of bacterial growth, including: - Vegetative growth refers to actively dividing cells as opposed to population growth. - Generation time is the time for a population to double in number through binary fission. - The four phases of bacterial growth in a culture are lag phase, log/exponential phase, stationary phase, and death phase. - Direct methods to measure bacterial growth include plate counts, filtration, and direct microscopic counts. Indirect methods include turbidity and dry weight.

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75 views32 pages

Notes

This document defines and describes various aspects of bacterial growth, including: - Vegetative growth refers to actively dividing cells as opposed to population growth. - Generation time is the time for a population to double in number through binary fission. - The four phases of bacterial growth in a culture are lag phase, log/exponential phase, stationary phase, and death phase. - Direct methods to measure bacterial growth include plate counts, filtration, and direct microscopic counts. Indirect methods include turbidity and dry weight.

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Mdm Aslizah Mohd Aris

Faculty of Applied Sciences

Bacterial Growth Definition

Vegetative growth

Microbial growth

Cells actively growing and dividing

Population growth as opposed to cell growth


Measured by total number of cells
Exponential due to binary fission reproduction
The increase in number of cells, not cell size

Generation time (or doubling time)

The time for an individual or a population to double

Binary fission
Budding

Binary Fission

Generation Time (Doubling Time)


time required for a cell to divide
most about 1 Hr. To 3 Hrs.
E. coli - 20 minutes
Mycobacterium tuberculosis - 24 Hrs.

E. coli - generation time of 20 min.


20 generations (about 7 hrs.)
1 million cells
30 generations ( about 10 hrs.)
1 billion cells
72 generations ( about 24 hrs.)
1 x 1021
1,000,000,000,000,000,000,00
0 cells

If 100 cells growing for 5 hours


produced 1,720,320 cells:

Bacillus cereus divides every 30


minutes. You inoculate a culture with
exactly 100 bacterial cells. After 3
hours, how many bacteria are present?

Answers:

Log end- log 100


0.301
Log end 2
0.301
11

6 = log end 2
0.301
Log end = 3.5
103.5= 6310

12

Lets say you have determined that your sample contains


6,400 bacterial cells. You know that it incubated 3
hours. How many generations have occurred?
Number of generations =

(log cells at end of incubation ) - (log cells at beginning of incubation)


0.301

(log 6400) - (log 100)


0.301
= (3.81 - 2)
0.301
= 6 generations
To calculate the generation time for a population:
60 min x hours
number of generations
In this example:
60 min x 3 hours
6 generations
= 30 minutes per generation

13

Direct methods
Plate counts
Filtration
MPN
Direct microscopic
count
Dry weight

Indirect methods
Turbidity
Metabolic activity
Dry weight

Plate counts: Perform serial dilutions of a sample

Figure 6.15, step 1

Inoculate Petri
plates from
serial dilutions

Figure 6.16

After incubation, count colonies on plates that have


25-250 colonies (CFUs)

Filtration

Direct microscopic count

Turbidity

Bacterial Growth

Four phases of growth in a culture


Lag phase
Log phase (or exponential phase)
Stationary phase
Death phase (or decline phase)

Will encompass several hours


During this time the organisms
grow in size, accumulate organic
matter and store large quantities of
chemical energy such as ATP for
biosynthesis

Microorganisms undergo rapid cell division


and fulfill their generation time.
The
population doubles during each
generation time
The population increases in size at a
logarithmic or exponential rate
Rapid cell growth (exponential growth)
population doubles every generation
microbes are sensitive to adverse conditions

antibiotics
anti-microbial agents

During the third phase of a populations history,


the rate of cell division decreases and the older
cells begin to die.
During this phase, the number of living cells in
the population remains constant.
Death rate = rate of reproduction
cells begin to encounter environmental stress
lack of nutrients
lack of water
not enough space
metabolic wastes
oxygen
pH
Endospores would form now

During the fourth phase, the death/decline


phase, the environment has become difficult
for living and the rate of cell death exceeds the
rate of cell division.
The population declines in numbers and the
population may completely die out if the
environmental stresses are not reversed.
Death rate > rate of reproduction
Due to limiting factors in the environment

1. You perform a serial dilution and determine that the original


number of cells in your sample was 20, 000. How many bacteria
will be present in 12 hours if the generation time is 15 minutes
(assume unlimited food and clean environment)?
2. You determine that a coconut cream pie has 5 million (3 x 106)
Staphylococcus aureus cells in it. You estimate that the food
preparer did not wash his hands and probably inoculated the
cream with 10 S. aureus. He also forgot to refrigerate it. If the
pie was made 6 hours ago, how many generations have
occurred? How long is each generation?
3. Using the generation time from problem 2, how many bacteria
would be present after 8 hours at room temperature?
32

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