2013.igem.

org

ITP 220 Food Microbiology

MICROBIAL GROWTH
L Nuraida, WP Rahayu and HD Kusumaningrum

Food Technology Bachelor Program

Department of Food Science and Technology,
Faculty of Agricultural Technology and Engineering
 TOPIC
Week Topic
1 Food Microbiology in Global Perspectives
2 Microbial Growth
3 Intrinsic and Extrinsic Factors of Microbial Growth
4 Computer Simulation Program: Microbial Growth
5 Introduction to Food Fermentation
6 Microbial Spoilage of Meat, Eggs, Milk, Seafood and Their
Products
7 Microbial Spoilage of Vegetables, Fruits, Cereal and Their
Products
Mid Exam
Department of Food Science and Technology
Bogor Agricultural University
LN
Sub Topics

2.1. Microbial Growth

2.2. Microbial Growth Kinetic

2.3. Microbial Growth Curve

2.4. Microbial Growth Measurement

Department of Food Science and Technology
Bogor Agricultural University
LN
 Learning Outcome

Learning Outcomes
1. Students are able to explain microbial growth in
general
2. Students are able to explain microbial growth,
kinetics, growth curve, and growth measurements
3. Students are able to determine the kinetics of
microbial growth
4. Students are able to calculate the rate of microbial
growth Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth

• Growth
– increase in the number of microbial cells
– increase in microbial mass
• Growth rate
– change in cell number or mass per unit time
• Generation
– interval for formation of two cells from one
• Generation time (doubling time)
– time required for formation of two cells
– vary widely among organisms
Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth

• Formation of two cells the size of its original cells:

– Cell continually increases in size until it is approximately
double
– Cell division occurs
• Large population cells develop quickly
– cells numbers in millions, hundred millions or billions
– difficult to handled such large numbers
• 1000 000 = 106 250 000 000 = 2.5 x 108
• 10 000 000 = 107
• 100 000 000 = 108
• 500 000 000 = 5.0 x 108
Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth

Bacterial division

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth
Data for population which doubles every 30 minutes
Time (h) Number of cells
0 1
0.5 2
1 4
1.5 8
2 16
2.5 32

10 1048576
Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth

Example of Microbial Growth

• If initial level of bacteria is 100, then after 10 h
the number of bacteria is
– 100 x 1048576 = 104857600 cell
• number of bacteria is written in one decimal, the
above number is becoming 1.0 x 108

Imagine if the initial number of bacteria is 105

Fresh milk arrived in milk processing plan is about

105 cell/mL
Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth

Reproduction of a Single Cells

Parameter

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Kinetic

Quantitative relationship of growth (exponential phase)

• Exponential Growth
– the number of cell increases by a constant factor
during each unit time period
– the rate of increase in the cell number is slow initially
but increases at an ever faster rate
– explosive increase in cell numbers
• Growth data are plotted on an arithmetic scale
– log10 value or semi-logarithmic graph

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Kinetic

Quantitative relationship of growth (exponential phase)

• dX/dt = kX
• ln X = ln Xo + k(t)
–where:
X = cell number at t time
Xo= cell number at time 0
k = growth rate constant
t = elapsed time
• Taking of antilogarithm
–X = Xo ekt Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth Kinetic
– Doubling of population has occurred when
• X/Xo = 2
–X = Xo ekt
–X/Xo = ekt
–2 = ek(t.gen) t.gen = generation time
–k = ln 2/t.gen
–k = 0.693/t.gen ; 1/t.gen = µ

–k = 0.693 µ or µ = k/0.693
Department of Food Science and Technology
µ = number of doubling per h or growth rate
Bogor Agricultural University
LN
Microbial Growth Kinetic

– Ln X - ln Xo = kt
– log X - log Xo = kt/2.303
– log X - log Xo = 0.693 µ t/2.303
– log X - log Xo = 0.301 µ t
• µ = (log Xt - log Xo)/0.301 t
• where µ = number of doubling per h or
growth rate

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Kinetic

 At initial time the number of bacteria is 103.

After 5 h the number of bacteria is 107.
Calculate the growth rate and generation time.

• µ = (log Xt - log Xo)/0.301 t

• µ = (log 107 - log 103)/0.301 t
• µ = 4/(0.301x5) = ………… (this is number of doubling
per h or growth rate)
• µ = 1/t. gen;
• t.gen = 1/ µ = …………. h (this is generation time)

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Curve
Growth data are plotted on an arithmetic scale:
log10 value or semilogarithmic graph

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Curve
The Growth Cycle of Populations

• Lag Phase
– when a microbial population is inoculated into a fresh
medium, growth usually does not begin immediately, but
only after a period of time
– lag is not seen when an exponentially growing culture is
inoculated into the same medium and the same conditions
– observed when a population is transferred from a rich
culture medium to a poorer one
• the cells must have complete complement of enzymes
Department of Food Science and Technology
Bogor Agricultural University
LN
 Microbial Growth Curve

• Exponential Phase (Logarithmic or Log Phase)

– the rates vary greatly
• in culture medium:
– generation time of Salmonella Typhi: 20 to 30 min.
– generation time of Mycobacterium tuberculosis: 12 -24 h
– the rate influenced by environmental conditions
(temperature, composition of culture medium) and
characteristic of organisms
• bacteria grow faster than eucaryotic microorganisms
• small eucaryotes grow faster than large one
Department of Food Science and Technology
Bogor Agricultural University
LN
Microbial Growth Curve

• Stationary Phase
– A single bacterial cell weight 1/1012 g
• If a single bacterium having generation time of
20 min. and continues to grow exponentially for 48 h.
– A single cell produces a population weighed
about 4 000 times the weight of the earth !!!!!
– But it never happens
– exponential growth ceases:
• essential nutrient of the culture used up
• some waste product of the microorganisms builds up
in the medium to an inhibitory level
Department of Food Science and Technology
Bogor Agricultural University
LN
 Microbial Growth Curve

• Stationary Phase
– no net decrease or increase in cell number
– no growth occurs
– But, many cell functions may continue,
including energy metabolism and biosynthesis
– certain microorganisms produce secondary
metabolites such as toxins, antibiotics

Department of Food Science and Technology

Bogor Agricultural University
LN
Microbial Growth Curve

• Death Phase
– total count (measured by direct microscopic count)
may remain constant
• In some cases death is accompanied by cell lysis
leading to decrease in the DMC
– viable count slowly decrease

• Lag phase, exponential phase, stationary phase

and death phase do not apply to individual cell
but only to population of cells

Department of Food Science and Technology

Bogor Agricultural University
LN
Measurement of Microbial Growth

• Measure:
– changes in number of cells or
– weight of cell mass
– turbidity of cell suspension
• Methods:
– Total cell count (DMC)
– Viable count
– Cell mass
– Turbidity
Department of Food Science and Technology
Bogor Agricultural University
LN
Measurement of Microbial Growth
 Total count (DMC)
 tedious but quick way of estimating microbial cell number
 Viable Count
 determine the number of cells in the sample capable of

forming colonies on a suitable agar medium

 also called plate count or colony count

 Methods:
 Spread plate

 Pour plate

Department of Food Science and Technology

Bogor Agricultural University
LN
Measurement of Microbial Growth

• Cell Mass
– Net weight measurement
• centrifuging the cells and weighing the pellet
• dry weight obtained by drying the centrifuge mass, usually
by placing it overnight at 100 – 105 oC
– dry weight usually 10-15 % of wet weight
• filtration of culture media on a membrane filter, the filter
is oven dried.
– Turbidity measurement
• expressed as unit absorbance
• used widely to follow the rate of growth of the culture
Thank You

Department of Food Science and Technology

Bogor Agricultural University
LN