7th week:

Chapter 5.
Microbial Growth
(미생물의 성장)
 Chapter 5

Chapter 6
(12th Edition)
Microbial Growth
(미생물의 성장)
 Cell Growth (세포 성장)
• Growth: increase in the number of
cells (미생물의 성장 – 세포수의 증가) Binary Fission in a Rod-Shaped
• Binary fission (이분법): cell elongation Prokaryote
following enlargement of a cell to
twice its minimum size
• Generation time (세대시간, doubling
time): time required for a population
of microbial cells to double
• During cell division each daughter
cell receives a chromosome and
sufficient copies of all other cell
constituents to exist as an
independent cell
Source: Brock Biology of Microorganisms 12th edition.
Figure 6.1
 Growth of Bacterial Populations
(세균 개체군의 성장)
• Most bacteria have shorter generation times than eukaryotic microbes
– Generation time is dependent on growth medium and incubation conditions
• Exponential growth (지수 성장)
– growth of a microbial population in which cell numbers double within a specific
time interval
– During exponential growth, the increase in cell number is initially slow but
increases at a faster rate
• Increase in cell number in an exponentially growing bacterial culture is a
geometric progression of the number 2
• Relationship exists between the initial number of cells present in a
culture and the number present after a period of exponential growth:
N = No2n
where N is the final cell number, No is the initial cell number, and n
is the number of generations during the period of exponential
growth
 Growth of Bacterial Populations
(세균 개체군의 성장)
• Generation time (g, 세대시간) of the exponentially growing population is
g = t/n
where t is the duration of exponential growth and n is the number of
generations during the period of exponential growth
• Specific growth rate (k, 비성장속도, 비성장률) is calculated as
k = 0.301/g
• Division rate (v) is calculated as
v = 1/g
Exponential phase
Nutrients and conditions are not limiting
growth = 2n or X = 2nX0
20
Where X0 = initial number of cells
X = final number of cells 21
n = number of generations
22

23

24

2n

Source: Environmental Microbiology 2nd edition.

Figure 3.5
 Microbial Growth Cycle (미생물의 성장 주기)
• Batch culture: a closed-system microbial culture of fixed volume
• Typical growth curve for population of cells grown in a closed system is
characterized by four phases
– Lag phase (준비기, 유도기)
– Exponential phase (지수성장기)
– Stationary phase (안정기)
– Death phase (사멸기)
• Lag phase
– Interval of time between when a culture is inoculated and when growth begins
• Exponential phase
– Cells in this phase are typically in the healthiest state
• Stationary phase
– Growth rate of population is zero
– Either an essential nutrient is used up or waste product of the organism
accumulates in the medium
Microbial Growth Cycle (미생물의 성장 주기)
Typical Growth Curve for a Bacterial Population

Source: Brock Biology of Microorganisms 12th edition.

Figure 6.10
 Continuous Culture (연속 배양)
• Continuous culture (연속배양) : an open-system microbial culture of
fixed volume
• Chemostat
– most common type of continuous culture device
– Both growth rate and population density of culture can be controlled
independently and simultaneously
• Dilution rate (희석률, 희석속도) : rate at which fresh medium is pumped in and
spent medium pumped out
• Concentration of a limiting nutrient (제한 영양물질의 농도)
– In a chemostat
• The growth rate is controlled by dilution rate
• The growth yield (cell number/ml) is controlled by the concentration of the
limiting nutrient
 Continuous Culture (연속 배양)
• Chemostat cultures are sensitive to the dilution rate and limiting nutrient
concentration
– Too high a dilution rate, the organism is washed out
– Too low a dilution rate, the cells may die from starvation
– Increasing limiting nutrient concentration results in greater biomass but same
growth rate
Steady-State (안정화 상태)
Schematic for Continuous Culture
Device (Chemostat)

Source: Brock Biology of Microorganisms 12th edition.

Figures 6.11 & 6.13
 Measuring Microbial Growth
(미생물 성장의 측정)
• Microscopic counts (현미경 계수)
– Measurements of total cell numbers
– Microbial cells can be enumerated by microscopic observations; simple but
results can be unreliable
• Limitations of microscopic counts
– Cannot distinguish between live and dead cells without special stains
– Small cells difficult to see and can be overlooked
– Precision is difficult to achieve
– A phase-contrast microscope is required if a stain is not used
– Cell suspensions of low density (< 106 cells/ml) hard to count
– Motile cells need to immobilized
– Debris in sample can be mistaken for cells
• A second method for enumerating cells in liquid samples is with a flow
cytometer
 Measuring Microbial Growth
(미생물 성장의 측정)
Direct Microscopic Counting Procedure

Source: Brock Biology of Microorganisms 12th edition.

Figure 6.14
 Measuring Microbial Growth
(미생물 성장의 측정)
• Viable cell counts (plate counts, 생균수, 평판계수)
– measurement of living, reproducing population
– To obtain the appropriate colony number, the sample to be counted should
always be diluted
– Two main ways to perform plate counts
• Spread-plate method
• Pour-plate method
– Plate counts can be highly unreliable when used to assess total cell numbers of
natural samples (e.g., soil and water)
– The Great Plate Anomaly
• direct microscopic counts of natural samples typically reveal far more
organisms than are recoverable on plates of any given culture medium
• Microscopic methods count dead cells whereas viable methods do not
• Different organisms in even a very small sample may have vastly different
requirements for resources and conditions in laboratory culture
 Measuring Microbial Growth
(미생물 성장의 측정)
Method for the Viable Count

Source: Brock Biology of Microorganisms 12th edition.

Figure 6.15
 Measuring Microbial Growth
(미생물 성장의 측정)
• Turbidity measurements are an indirect but very rapid and useful
method of measuring microbial growth
– Most often measured with a spectrophotometer and measurement referred to
as optical density (O.D., 흡광도)
– To relate a direct cell count to a turbidity value, a standard curve must first be
established

• Turbidity measurements
– Quick and easy to perform
– Typically do not require destruction or significant disturbance of sample
– Sometimes problematic (e.g., microbes that form clumps or biofilms in liquid
medium)
 Temperature and Microbial Growth
(온도와 미생물 성장)
• Temperature is a major environmental factor controlling microbial
growth
• Cardinal temperatures: the minimum (최저성장), optimum (최적성장온도),
and maximum (최고성장) temperatures at which an organism grows
• Microorganisms can be classified into groups by their growth
temperature optima
– Psychrophile: low temperature (저온성)
– Mesophile: midrange temperature (중온성)
– Thermophile: high temperature (고온성)
– Hyperthermophile: very high temperature (초고온성)
• Mesophiles (중온성)
– organisms that have midrange temperature optima
– found in warm-blooded animals, terrestrial and aquatic environments of
temperate and tropical latitudes
 Temperature and Microbial Growth
(온도와 미생물 성장)
• Psychrophiles (저온성)
– Organisms with cold temperature optima; the most extreme representatives
inhabit permanently cold environments
• Psychrotolerant (저온내성)
– Organisms that can grow at 0ºC but have optima of 20ºC to 40ºC; more widely
distributed in nature than psychrophiles
• Molecular Adaptations to Psychrophily
– Production of enzymes that function optimally in the cold
– features that may provide more flexibility
– Transport processes function optimally at low temperatures due to
modifications of cytoplasmic membranes
• High unsaturated fatty acid content
 Temperature and Microbial Growth
(온도와 미생물 성장)
• Thermophiles (고온성)
– organisms with growth temperature optima between 45ºC and 80ºC
• Hyperthermophiles (초고온성)
– organisms with optima greater than 80°C (최적성장온도가 80℃ 이상)
– Inhabit hot environments including boiling hot springs and seafloor
hydrothermal vents that can have temperatures in excess of 100ºC
• Studies of thermal habitats have revealed
– Prokaryotes are able to grow at higher temperatures than eukaryotes
– Organisms with the highest temperature optima are Archaea
– Nonphototrophic organisms can grow at higher temperatures than phototrophic
organisms
 Temperature and Microbial Growth
(온도와 미생물 성장)
• Molecular Adaptations to Thermophily
– Enzyme and proteins function optimally at high temperatures; features that
provide thermal stability
• Critical amino acid substitutions in a few locations provide more heat-
tolerant folds
• An increased number of ionic bonds between basic and acidic amino acids
resist unfolding in the aqueous cytoplasm
• Production of solutes help stabilize proteins
– Modifications in cytoplasmic membranes to ensure heat stability
• Bacteria have lipids rich in saturated fatty acids
• Archaea have lipid monolayer rather than bilayer
• Hyperthermophiles and produce enzymes widely used in industrial
microbiology
– E.g., Taq polymerase; used to automate the repetitive steps in the polymerase
chain reaction (PCR) technique
 Temperature and Microbial Growth
(온도와 미생물 성장)
Temperature and Growth Relations in Different Classes

Source: Brock Biology of Microorganisms 12th edition.

Figure 6.19
 pH and Microbial Growth
(pH와 미생물 성장)
• The pH of an environment greatly affects microbial growth
• Neutrophiles (호중성)
– Some organisms have evolved to grow best at low or high pH, but most
organisms grow best between pH 6 and 8
• Acidophiles (호산성)
– organisms that grow best at low pH (< 6)
– Some obligate acidophiles; membranes destroyed at neutral pH
– Stability of cytoplasmic membrane critical
• Alkaliphiles (호염기성)
– organisms that grow best at high pH (> 9)
– Some have sodium motive force rather than proton motive force
• The internal pH of a cell must stay relatively close to neutral even
though the external pH is highly acidic or basic
– Internal pH has been found to be as low as 4.6 and high as 9.5 in extreme
acido- and alkaliphiles, respectively
– Microbial culture media typically contain buffers to maintain constant pH
 NaCl and Microbial Growth
(염분농도와 미생물 성장)
• Water activity (aw)
– water availability; expressed in physical terms
– Defined as ratio of vapor pressure of air in equilibrium with a substance or
solution to the vapor pressure of pure water
• Typically, the cytoplasm has a higher solute concentration than the
surrounding environment, thus the tendency is for water to move into
the cell (positive water balance)
• When a cell is in an environment with a higher external solute
concentration, water will flow out unless the cell has a mechanism to
prevent this
 NaCl and Microbial Growth
(염분농도와 미생물 성장)
• Halophiles (호염성)
– organisms that grow best at reduced water potential; have a specific
requirement for NaCl
• Extreme halophiles (극호염성)
– organisms that require high levels (15–30%) of NaCl for growth
• Halotolerant (염분내성)
– organisms that can tolerate some reduction in water activity of environment but
generally grow best in the absence of the added solute
• Mechanisms for combating low water activity in surrounding
environment involve increasing the internal solute concentration by
– Pumping inorganic ions from environment into cell
– Synthesis or concentration of organic solutes
• compatible solutes: compounds used by cell to counteract low water
activity in surrounding environment
 Oxygen and Microbial Growth
•
(산소와 미생물
Aerobes (호기성) : require oxygen to live
성장)
• Anaerobes (혐기성) : do not require oxygen and may even be killed by
exposure
• Facultative organisms: can live with or without oxygen
• Aerotolerant anaerobes: can tolerate oxygen and grow in its presence
even though they cannot use it
• Microaerophiles (미호기성): can use oxygen only when it is present at
levels reduced from that in air
• Several toxic forms of oxygen can be formed in the cell
– Single oxygen
– Superoxide anion
– Hydrogen peroxide
– Hydroxyl radical
• Enzymes are present to neutralize most of these toxic oxygen species