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Binary fission
The growth cycle
Measurements of microbial growth
Environmental effects on microbial growth: temperature, pH,
water activity, oxygen
The general process of binary fission in a rod-shaped prokaryote. For simplicity, the nucleoid is depicted as a single circle in green.
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FtsI activity is blocked by penicillin
Phase contrast
Nucleoid stain
FtsZ stain
The FtsZ ring and cell division. Cell division and chromosome replication are coordinately regulated, and the Fts
proteins are the keys to these processes. Fts proteins interact to form a division apparatus in the cell called the
divisome.
Cell wall synthesis in gram-positive Bacteria. (a) Localization of new cell wall synthesis during cell division. In cocci, new cell
wall synthesis (shown in green) is localized at only one point. The FtsZ ring defines the cell division plane.
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Typical growth curve for a bacterial population in a batch culture (closed system). There is usually a lag phase, then exponential
growth commences. As essential nutrients are depleted or toxic products build up, growth ceases, and the population enters the
stationary phase. If incubation continues, cells may begin to die (the death phase).
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Compare the rate of cell production 22
between 0-0.5 h and 4-4.5 h 23
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N = N02n
N = Final cell number
N0 = Initial cell number
n = Number of generations
g = t/n
g = Generation time (doubling time)
t = Time of exponential growth
The rate of growth of a microbial culture. (a) Data for a population that doubles every 30 min. (b) Data plotted on an
arithmetic (left ordinate) and a logarithmic (right ordinate) scale.
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The Mathematics of Exponential Growth
Method of estimating the generation times (g) of exponentially growing populations with generation times of (a) 6 h and (b)
2 h from data plotted on semilogarithmic graphs. The slope of each line is equal to 0.301/g and n equals the number of
generations that have occurred in the time, t.
Growth kinetics
Growth rate constant - number of generations that occur per unit time - (expressed as k or µ; h -1 ):
(1) k = (ln N 2 - ln N1)/(t2 - t 1) where N2 and N 1 = cells ml -1 at time t2 and t1 (in h)
(2) Convert (1) to log: k = (log N2 - log N1 ) (2.303)/(t2 - t 1)
Generation time - time required for the cell population to double - (expressed as g; h):
(3) g = (ln 2)/k
Example I: A bacterial culture in exponential growth contained 10 4 cells/ml at time zero and 10 8 cells/ml at time 4 h. What
is the growth rate constant (k)?
Example II: E. coli grew exponentially from 5 x 10 5 cells/ml to 3.5 x 107 cells/ml in 5 h. Its generation time was 40 min.
Was there a lag phase?
From (1) and (3) (t2 - t 1) = (ln N2 - ln N1)/k = g (ln N2 - ln N1)/ln 2 = 40 [ln (3.5 x 107) - ln (5 x 105)]/0.693 = 245 min
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Direct microscopic counting procedure using
the Petroff-Hausser counting chamber.
Method not suitable for cell suspensions of low density or motile cells
Total cell counts (done microscopically) measure the total number of cells in a population
Two methods of performing a viable count (plate count). In either case the sample must usually be diluted before plating.
Cell counts (plate counts) measure only the living, reproducing population.
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Procedure for viable counting using serial dilutions of the sample and the pour plate method.
Plate counts reveal lower numbers of cells that direct microscopic counts
Why?
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What if an organism does not grow on solid medium?
Turbidity measurements of microbial growth. (a) Measurements of turbidity are made in a spectrophotometer or photometer. The
photocell measures incident light unscattered by cells in suspension and gives readings in optical density or photometer units.
Turbidity measurements of microbial growth. (b) Typical growth curve data obtained in Klett units or optical density (OD) for
two organisms growing at different growth rates. For practice, calculate the generation time ( g) of the two cultures using the
formula n = 3.3 (log N – log N0) where N and N 0 are two different Klett values taken between a time interval t . Which organism is
growing faster, A or B? (c) Relationship between cell number or dry weight and turbidity readings. Note that the one-to-one
correspondence between these relationships breaks down at high turbidities.
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Batch cultures vs. continuous cultures
Batch culture
µ= 0.48 h -1
Cells ml-1
Time (h)
Schematic for a continuous culture device (chemostat). In such a device, the population density is controlled by the
concentration of limiting nutrient in the reservoir, and the growth rate is controlled by the flow rate. Both parameters can be set
by the experimenter. Chemostats are a means of maintaining cell populations in exponential growth for long periods.
Relationship between nutrient concentration, growth rate (green curve), and growth yield (red curve) in a batch culture
(closed system). At low nutrient concentrations both growth rate and growth yield are affected.
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Steady-state relationships in the chemostat. The dilution rate is determined from the flow rate and the volume of the
culture vessel.
Narrow range
Wide range
Relation of temperature to growth rates of a typical psychrophile, a typical mesophile, a typical thermophile, and two
different hyperthermophiles.
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Effect of temperature on growth rate and the molecular consequences for the cell. The three cardinal
temperatures vary by organism.
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Adaptations to High Temperatures
Stability of monomers: ATP and NAD+ have half-life < 30 min at 120°C
Protein stability: Increased flexibility
The pH scale. Note that although some microorganisms can live at very low or very high pH, the cell’s internal pH
remains near neutrality.
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aw: ratio of the vapor pressure of the air in equilibrium with a substance to the vapor pressure of pure water
Effect of sodium ion concentration on growth of microorganisms of different salt tolerances or requirements. The optimum NaCl
concentration for marine microorganisms such as V. fischeri is about 3%; for extreme halophiles, it is between 15 and 30%,
depending on the organism.
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Water activity becomes limiting to an organism when the dissolved solute concentration in its environment increases.
To counteract this situation, organisms produce or accumulate intracellular compatible solutes that maintain the cell
in positive water balance.
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Aerobic (a), anaerobic (b), facultative (c), microaerophilic (d), and aerotolerant (e) anaerobe growth, as revealed by the
position of microbial colonies (depicted here as black dots) within tubes of thioglycolate broth culture medium.
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Several toxic forms of oxygen can be formed in the cell…
Four-electron reduction of O 2 to water by stepwise addition of electrons. All the intermediates formed are reactive and
toxic to cells except for water, of course.
Enzymes that destroy toxic oxygen species. (a) Catalases and (b) peroxidases are porphyrin-containing proteins, although some
flavoproteins may consume toxic oxygen species as well. (c) Superoxide dismutases are metal-containing proteins, the metals being
copper and zinc, manganese, or iron. (d) Combined reaction of superoxide dismutase and catalase. (e) Superoxide reductase
catalyzes the one electron reduction of O 2- to H2O2 using reduced cytochrome c as the electron donor.
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Method for testing a microbial culture for the presence of catalase. A heavy loopful of cells from an agar culture was mixed on a
slide with a drop of 30% hydrogen peroxide. The immediate appearance of bubbles is indicative of the presence of catalase. The
bubbles are O 2 produced by the reaction H2O2 + H2O2 → 2 H 2O + O2.
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