07 Lecture 1

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Chapter 7
Microbial Growth and

Reproduction
1
Reproductive Strategies
• the reproductive strategies of
eukaryotic microbes
– asexual and sexual, haploid or diploid
• Bacteria and Archaea
– haploid only, asexual - binary fission,
budding, filamentous
– all must replicate and segregate the
genome prior to division
2
Figure 7.1
3
Figure 7.2
4
Bacterial Cell Cycle

• cell cycle is sequence of events from
formation of new cell through the
next cell division
– most bacteria divide by binary fission
• two pathways function during cycle
– DNA replication and partition
– cytokinesis
5
Chromosome Replication and

Partitioning - 1
• most bacterial chromosomes are circular
• single origin of replication – site at which
replication begins
• terminus – site at which replication is
terminated, located opposite of the origin
• replisome – group of proteins needed for DNA
synthesis
• DNA replication proceeds in both directions
from the origin
• origins move to opposite ends of the cell
6
Figure 7.3
7
Chromosome Partitioning
• replisome pushes, or condensation of,
daughter chromosomes to opposite ends
• MreB (murein cluster B) – an actin
homolog, plays role in determination of
cell shape as spiral inside cell periphery,
and chromosome segregation
– new origins associate with MreB tracks
– if MreB is mutated, chromosomes do not
segregate
8
Figure 7.4
9
Plasmid Segregation
• plasmids replicate independently and
carry proteins necessary for segregation
• E. coli R1 plasmid produces three
proteins essential for its inheritance
– ParM – similar to MreB, actin homolog
forms long filaments
– ParR (repressor) and ParC (centromere-
like) both bind to origins and link to ParM
– ParM filaments elongate and separate
plasmids to opposite ends of cell
10
Figure 7.5
11
Cytokinesis - Septation
• septation – formation of cross walls
between daughter cells
• several steps
– selection of site for septum formation
– assembly of Z ring
– linkage of Z ring to plasma membrane (cell
wall)
– assembly of cell wall synthesizing machinery
– constriction of cell and septum formation
12
Z Ring Formation - Role in

Septation
• protein FtsZ
– tubulin homologue, found in most bacteria and
archaea
– polymerization forms Z ring, filaments of meshwork
• MinCDE system in E. coli limits the Z ring to the
center of the cell
– MinC, MinD, MinE oscillate from one side of cell to
other
– link Z ring to cell membrane
– Z ring constricts and cell wall synthesis of septal wall
13
Figure 7.6
14
Figure 7.7
15
Table 7.1
16
Cellular Growth and

Determination of Cell Shape
• determined by peptidoglycan synthesis
in bacteria
– penicillin binding proteins (PBPs) – link
peptidoglycan strands and catalyze
controlled degradation for new growth
– autolysins – PBP enzymes that degrade
peptidoglycan and site new units added
17
Figure 7.8
18
Cell Wall Growth and Cell

Shape Determination
• cocci divisome - new peptidoglycan forms
only at the central septum
– FtsZ determines site of cell wall growth
– FtsZ may recruit PBPs for synthesis of
septum
• rods are similar but elongate prior to
septation
– MreB determines cell diameter and
elongation as Z ring forms in center
19
Figure 7.9
20
Cell Wall Growth and Cell

Shape Determination
• vibrio (comma-shaped bacteria)
– FtsZ – forms Z ring
– MreB – helical polymerization throughout
cell
– crescentin – intermediate filament
homologue cytoskeletal protein
• localizes to short, curved side of cell
• asymmetric cell wall synthesis forms curve
21
Figure 7.10
22
Growth
• increase in cellular constituents
that may result in:
– increase in cell number
– increase in cell size
• growth refers to population
growth rather than growth of
individual cells
23
The Growth Curve

• observed when microorganisms are
cultivated in batch culture
– culture incubated in a closed vessel with a
single batch of medium
• usually plotted as logarithm of cell
number versus time
• has four distinct phases
– lag, exponential, stationary, senescence, and
death
24
Figure 7.11
25
Lag Phase
• cell synthesizing new components
– e.g., to replenish spent materials
– e.g., to adapt to new medium or other
conditions
• varies in length
– in some cases can be very short or even
absent
26
Exponential Phase
• also called log phase
• rate of growth and division is
constant and maximal
• population is most uniform in terms
of chemical and physical properties
during this phase
27
Balanced Growth
• during log phase, cells exhibit
balanced growth
– cellular constituents manufactured at
constant rates relative to each other
28
Unbalanced Growth
• rates of synthesis of cell components
vary relative to each other
• occurs under a variety of conditions
– change in nutrient levels
• shift-up (poor medium to rich medium)
• shift-down (rich medium to poor medium)
– change in environmental conditions
29
Figure 7.12
30
Stationary Phase
• closed system population growth
eventually ceases, total number of
viable cells remains constant
– active cells stop reproducing or
reproductive rate is balanced by death
rate
31
Possible Reasons for

Stationary Phase
• nutrient limitation
• limited oxygen availability
• toxic waste accumulation
• critical population density reached
32
Stationary Phase and

Starvation Response
• entry into stationary phase due to
starvation and other stressful conditions
activates survival strategy
– morphological changes
• e.g., endospore formation
– decrease in size, protoplast shrinkage, and
nucleoid condensation
– RpoS protein assists RNA polymerase in
transcribing genes for starvation proteins
33
Starvation responses
Bacteria in culture may inter into stationary phase in response
to starvation. This may lead to ;
– morphological changes; e.g., endospore formation
– decrease in size, protoplast shrinkage, and nucleoid condensation
– production of starvation proteins that makes the cell more resistant to
damage by different ways;
• Increase peptidoglycan cross-linking and cell wall strength.
• Increase in the production of DNA binding proteins to protects DNA
• Chaperon proteins protect against protein denaturation and renature
damaged proteins
• As a result of these measures, starved cells become hard to
kill, they become resistant to starvation, resistant to
chemicals such as chlorine, damaging temperatures and
oxidative stress. This leads to;
• long-term survival
• increased their virulence
34
Death Phase
It was argued that when cells are in the decline phase, they are dead,
however, there is a debate over this statement. There are two alternative
hypotheses explaining the phenomenon;
– Cells are Viable But Not Culturable (VBNC)
• Cells alive, but dormant. Cells were genetically
programmed to survive. They will not be grown under
laboratory conditions but in animal passage they might
grow again.
– Programmed cell death
• Fraction of the population genetically programmed to die (commit
suicide). Those will leak their nutrients to be consumed by other
surviving bacteria that will not lyse and might grow later when
cells are passed through an animal.
35
Loss of Viability
Figure 6.8 In all cases, the survivors will not be able to grow under
36
regular laboratory conditions.
Loss of viability
• A; cells leave stationary phase due to starvation and
accumulation of toxic waste and inter decline phase.
• B; it is believed that a fraction of microbes go into

programmed cell death and nutrients released from dead cells
are consumed by other living cells.
• C; some part of the culture inters the state of viable but non-
culturable due to starvation. They can only grow through
passage through animals.
37
Prolonged Decline in Growth
Why it happen?
• bacterial population continually evolves as
they are better able to use released nutrients
upon the death of other cells.
• process marked by successive waves of
genetically distinct variants with more
capability of survival
• natural selection occurs
38
Prolonged Decline in Growth
Prolonged growth experiments reveal that an exponential decline
phase is some times replaced by gradual decline in the number
of culturable cells which can last for months or years. The most
strong cells will utilize nutrients released from dead cells and
tolerate toxic waste. i.e natural selection occurs
• bacterial population continually evolves
• process marked by successive waves of genetically distinct
variants
Figure 6.9
39
Measurement of Growth Rate and Generation Time
Figure 7.16
40
Table 7.2
41
Exponential microbial
growth as drawn from
data in table 7.2.
population is Logarithmic
doubling every
generation
Direct plot
Figure 7.15
42
Figure 7.17; generation time

determination
43
Table 7.3
44
Measurement of
Microbial Growth
• can measure changes in number of
cells in a population
• can measure changes in mass of
population
45
Measurement of Cell Numbers
• Most obvious method is direct cell counts
– Counting chambers; Petroff-Hausser counting chamber
can be used for counting prokaryotic cells. While
hemocytometers can be used for counting both prokaryotic
and eukaryotic cells.
– Electronic counters; such as Coulter counters are used for
direct counting of protists and yeast or cells can be counted
by flow cytometer.
– Membrane filters; used mainly for counting bacteria from
large liquid volumes.
• Viable cell counts
– plating methods
– membrane filtration methods
46
Counting Chambers
• easy,
inexpensive, and Figure 7.18
quick
• useful for
counting both
eukaryotes and
prokaryotes
• cannot
distinguish living
from dead cells
47
Direct Counts on Membrane

Filters
• cells filtered through special membrane
that provides dark background for
observing cells
• cells are stained with fluorescent dyes
• useful for counting bacteria
• with certain dyes, can distinguish living
from dead cells
48
Flow Cytometry
• microbial suspension forced through
small orifice with a laser light beam
• movement of microbe through orifice
impacts electric current that flows
through orifice
• instances of disruption of current are
counted
• specific antibodies can be used to
determine size and internal complexity
49
Viable Counting Methods

• spread and pour plate techniques
– diluted sample of bacteria is spread over
solid agar surface or mixed with agar and
poured into Petri plate
– after incubation the numbers of organisms
are determined by counting the number of
colonies multiplied by the dilution factor
– results expressed as colony forming units
(CFU)
50

• membrane filter technique
– bacteria from aquatic samples are
trapped on membranes of known pore
size
– membrane soaked in culture media
– colonies grow on membrane
– colony count determines number of
bacteria in original sample
51
Figure 7.19
52
Figure 7.20
53

• if microbe cannot be cultured on
plate media
• dilutions are made and added to
suitable media
• turbidity determined to yield the
most probable number (MPN)
54
Measurement of Cell Mass
An increase in the cell growth is accompanied by an increase in
cell mass.
Dry weight is the most direct method of cell mass estimation
– time consuming and not very sensitive
• Measuring the quantity of a particular cell constituent
– e.g., protein, DNA, ATP, or chlorophyll
– useful if amount of substance in each cell is constant
• Turbidometric measures (light scattering) of cultures
– Amount of scattered light is directly proportional to the
biomass of the cells. High absorbance means low
transmittance = high microbial growth
– quick, easy, and sensitive
55
Figure 7.21
56
The Continuous Culture of Microorganisms
Growing bacteria in a flask is a closed system as no addition of
new nutrients and no removal of waste therefore the log phase
only stays for short period of time.
• Growth in an open system means

– continual provision of nutrients
– continual removal of wastes and toxins
• This continuity maintains cells in log phase at a constant
biomass concentration for extended periods
• achieved using a continuous culture system (i.e fermenter)
57
The Chemostat
• Rate of incoming fresh

medium = rate of removal
of exhausted medium
from vessel
• An essential nutrient is in
limiting quantities which
determines the final
growth rate
58 Figure 6.16
Dilution Rate and Microbial Growth
dilution rate D, is the rate at which medium flows through vessel relative to vessel
size. F is the flow rate, v is the vessel volume. D = f/v
59
• Dilution rate reflects the amount of new nutrients added to the
media, so at first, when dilution rates are low the
microorganisms grow well because of the accumulation of the
nutrients but soon the essential nutrient will be depleted.
• Dilution rate increase means more nutrition so the generation

time decreases i.e growth increases and the biomass
maintained high at a wide dilution rates.
• This goes up to a level were the dilution rate is too high, so at

this time, nutrient concentration keeps increasing and the
generation time keeps decreasing.
• But the biomass will decrease sharply because this high

increase in dilution rates (which will be at some time greater
than the maximum growth rate) will wash out the bacteria
from the culture vessel and this will sharply decrease the
biomass.
60
The Turbidostat
This is another type of continues growth vessel but with a

photocell that regulates the flow rate of media through the
vessel to maintain a predetermined turbidity or cell density.
This system differs from the chemostat in the following:

– dilution rate varies
– no limiting nutrient
– turbidostat operates best at high dilution rates while chemostat does
not operate well at high dilution rates.
61
Importance of continuous culture methods
Industrial
1. Constant supply of cells in exponential phase growing at a
known rate. This could be beneficial if the organism is used
for vaccine production.
2. Food and industrial microbiology benefits. i.e production of
nutrients such as vitamins or certain proteins that is used as
drugs …etc.
Research
1. Study of microbial growth at very low nutrient
concentrations, close to those present in natural environment
2. Study of interactions of microbes under conditions resembling
those in aquatic environments
62
The Influence of Environmental Factors on
Growth
• Most organisms grow in fairly moderate environmental
conditions however some procaryotes can live in very harsh
environments. Example;
• Extremophiles
– grow under harsh conditions that would kill most other
organisms. e.g some bacteria can live under 1.5 miles
below earth surface, no O2 and temperature > 60˚ C.
– Table 7-4 summarizes the way in which microorganisms

are categorized in response to environmental factors.
63
Table 7.4
64
Solutes and Water Activity

• changes in osmotic concentrations in the
environment may affect microbial cells
– hypotonic solution (lower osmotic
concentration)
• water enters the cell
• cell swells may burst
– hypertonic (higher osmotic concentration)
• water leaves the cell
• membrane shrinks from the cell wall
(plasmolysis) may occur
65
Solutes and Water Activity…con
• Water activity (aw) or water potential
– amount of water available to organisms
– reduced by interaction with solute molecules (osmotic
effect)
higher [solute]  lower aw
– reduced also by adsorption to surfaces (matrix effect)
It is important that bacteria are able to respond to changes in
their osmotic concentrations in their environment. Example
microbes in hypotonic environment can reduce osmotic
concentrations in their cytoplasm by the use of inclusion
bodies (collecting the small proteins in large bulk) or some
other organisms will stretch the pores in their membranes and
allow solutes to leave to prevent water from accumulating
inside. Or some protists use contractile vacuoles to expel water
outside such as the Paramesium.
66
Water Activity (aw)
• water activity of a solution can be expressed quantitatively
and is equal to 1/100 of the relative humidity of solution.
• also equal to ratio of solution’s vapor pressure (Psoln) to that
of pure water (Pwater)
• Aw = Psoln/ Pwater
Aw of a solution can be measured by sealing the solution in a

chamber and measure its relative humidity (RH) after the
system has come to equilibrium. If RH reads 95 then Aw
would be 0.95.
Aw is inversely related to osmotic pressure.
67
Microbes Adapt to Changes in

Osmotic Concentrations
• reduce osmotic concentration of
cytoplasm in hypotonic solutions
– mechanosensitive (MS) channels in plasma
membrane allow solutes to leave
• increase internal solute concentration
with compatible solutes to increase their
internal osmotic concentration in
hypertonic solutions
– solutes compatible with metabolism and
growth
68
Extremely Adapted Microbes

• halophiles
– grow optimally in the presence of NaCl or
other salts at a concentration above about
0.2M
• extreme halophiles
– require salt concentrations of 2M and 6.2M
– extremely high concentrations of potassium
– cell wall, proteins, and plasma membrane
require high salt to maintain stability and
activity
69
Osmotolerant organisms
• Some microorganisms grow over wide ranges of water activity
and keep the osmotic concentrations above that of the habitat
by using
• Compatible solutes to increase their internal osmotic
concentration
– solutes that are compatible with metabolism and growth i.e
they do not interfere with metabolism. Example;
– Many microorganisms increase their internal osmotic
concentration in hypertonic environment by producing;
choline, betaine, proline, glutamic acid and other amino
acids and have some elevated levels of potassium ions.
• Some have proteins and membranes that require high solute
concentrations for stability and activity those are called
halophiles.
70
Effects of NaCl on microbial growth
• Halophiles
– grow optimally at >0.2 M
• Extreme halophiles
– require >2 M and saturation
up to 6.2 M. example;
The archaeon Halobcterium can be
isolated form the Dead Sea or
the salt lake in Utah. Those
organisms accumulate
enormous amounts of potassium
(up to 4-7 M) to remain
hypertonic to their
environment. In addition,
plasma membranes and cell
wall have high amounts of NaCl
that is necessary for their
function.
71 Figure 7-24
pH
• measure of the Figure 7.25
relative acidity
of a solution
• negative
logarithm of
the hydrogen
ion
concentration
72
pH
• Acidophiles
– growth optimum between pH 0 and pH 5.5. Most fungi
prefer pH 4-6
• Neutrophiles
– growth optimum between pH 5.5 and pH 7. most bacteria
and protists.
• Alkalophiles
– growth optimum between pH 8.5 and pH 11.5
– Drastic changes in pH disrupts the plasma membranes of

bacteria and might inhibit the activity of enzymes and
transport proteins.
73
pH
Microorganisms respond to external pH changes using
different mechanisms;
– Most acidophiles and alkalophiles maintain an internal pH
near neutrality
• Their plasma membrane is impermeable to protons
– Some bacteria synthesize proteins that provide protection

• e.g., chaperon proteins such as acid-shock proteins or heat shock
proteins. These prevent acid denaturation and aid in their
refolding.
• a proton translocating ATPase contribute to protecting the bacteria
from acidic media by making more ATP or pumping protons out of
the cell
74
pH
• most microbes maintain an internal pH near
neutrality
– the plasma membrane is impermeable to proton
– exchange potassium for protons
• acidic tolerance response
– pump protons out of the cell
– some synthesize acid and heat shock proteins that
protect proteins
• many microorganisms change the pH of their
habitat by producing acidic or basic waste
products
75
Effect of temperature
• The most important factor influencing the effect of
temperature is the temperature sensitivity of the enzyme-
catalyzed reactions with the speed of the enzyme reactions
double for every 10° C rise. Above certain temperatures the
speed slows and very high temperature become lethal as this
temperature will affect both structure and function aspects
of the microbe
• Very low temperatures solidify the bacterial membranes

thus affecting their function but not the structure.
• Organisms therefore have characteristic temperature -

dependence with distinct cardinal temperatures.
76
Temperature
• Organisms exhibit
distinct cardinal growth
temperatures
– minimal
– maximal
– Optimal
• These cardinal
temperatures are not
fixed, i.e based on
certain changes in the
media they might be
changed. Table 6-5
shows some of these
Temperatures.
77 Figure 6.20
Classes of organisms based on temperature
• Psychrophiles; grow well at 0 and have optimum at about 15 ° C
and maximum at 20 C .The cell membranes of the bacteria living
in this environment contains high amount of unsaturated fatty
acids that keeps the membrane semi-fluid.
• Psychrotrophs or facultative psycrophiles; grow at 0-7° C with
optimum between 20-30 ° C and maximum at about 35 ° C. this
type of bacteria and fungi are major food spoilage organisms.
• Mesophiles; are microorganisms with growth optimum 20-45 °

C. Minimum T is 15-20 ° C and maximum 45 ° C. Most
microbes fall in this category and of course almost all human
pathogens fall in this category i.e 37 ° C.
78
Temperature Ranges for

Microbial Growth (summary)
• psychrophiles – 0o C to 20o C
• psychrotrophs – 0o C to 35o C
• mesophiles – 20o C to 45o C
• thermophiles – 55o C to 85o C
• hyperthermophiles – 85o C to 113o C
79
Table 7.5
80
Figure 7.27
81
Adaptations of Thermophiles
• protein structure stabilized by a variety
of means
– e.g., more H bonds
– e.g., more proline
– e.g., chaperones
• histone-like proteins stabilize DNA
• membrane stabilized by variety of means
– e.g., more saturated, more branched and
higher molecular weight lipids
– e.g., ether linkages (archaeal membranes)
82
Oxygen Concentration
• growth in oxygen correlates with
microbes energy conserving
metabolic processes and the electron
transport chain (ETC) and nature of
terminal electron acceptor
83
Oxygen and Bacterial Growth

• aerobe
– grows in presence of atmospheric oxygen
(O2) which is 20% O2
• obligate aerobe – requires O2
• anaerobe
– grows in the absence of O2
• obligate anaerobe
– usually killed in presence of O2
84
Oxygen and Bacterial Growth
• microaerophiles
– requires 2–10% O2
• facultative anaerobes
– do not require O2 but grow better in its
presence
• aerotolerant anaerobes
– grow with or without O2
85
Figure 7.28
86
Oxygen Concentration
need prefer ignore oxygen is < 2 – 10%

oxygen oxygen oxygen toxic oxygen
87 Figure 6.22
Classification of organisms based on O2
requirements
• Obligate aerobes; contain SOD and Catalase and can not live
without O2. Oxygen serves as the terminal electron acceptor
for electron transport chain. It participates in sterol and fatty
acid synthesis in multicellular eukaryotes.
• Facultative anaerobes; contain SOD and catalase. do not
require O2 for growth but grow better in its presence.
• Aerotolerant; contains SOD but not catalase. Ignores the
presence of O2 and grow equally well whether its present or
not.
• Strict or obligate anerobe; does not contain SOD or catalase.
Can not live in its presence. Thus they obtain their energy
through fermentation.
• Microaerophiles; contain SOD and low levels of catalase.
Need 2-10% O2 not regular 20 %.
88
Basis of Different Oxygen

Sensitivities
• oxygen easily reduced to toxic reactive
oxygen species (ROS)
– superoxide radical
– hydrogen peroxide
– hydroxyl radical
• aerobes produce protective enzymes
– superoxide dismutase (SOD)
– catalase
– peroxidase
89
Strict Anaerobic Microbes

• all strict anaerobic microorganisms lack
or have very low quantities of
– superoxide dismutase
– catalase
• these microbes cannot tolerate O2
• anaerobes must be grown without O2
– work station with incubator
– gaspak anaerobic system
90
Figure 7.29; anaerobic work station
91
Figure 7.30
92
Pressure
• microbes that live on land and water
surface live at 1 atmosphere (atm)
• some Bacteria and Archaea live in
deep sea with very high hydrostatic
pressures
93
Pressure
• barotolerant
– adversely affected by increased
pressure, but not as severely as
nontolerant organisms
• barophilic (peizophilic) organisms
– require or grow more rapidly in the
presence of increased pressure
– change membrane fatty acids to adapt
to high pressures
94
The Electromagnetic Spectrum
Figure 7;31
95
Radiation
Radiation Damage
• Ionizing radiation; is very harmful to microorganisms. It has
very short wavelength and high energy.
• There are two major forms of ionizing radiation;
– x rays and gamma rays. Low levels of these radiation will
cause mutations  death
– Radiation disrupts chemical structure of many molecules,
including DNA. It destroys;
– Hydrogen bonds and oxidize double bonds.
– Destroys ring structures and polymerize some molecules
• Oxygen enhances these changes by producing hydroxyl fee radicals
– damage may be repaired by DNA repair mechanisms
however, higher levels are lethal. Therefore,
– Ionizing radiation is used for sterilization.
– Deinococcus radiodurans
96 • extremely resistant to DNA damage
Radiation Damage…
• Ultraviolet (UV) radiation; can kill all types of
microorganisms due to its short wavelength (10-400) and high
energy with the most lethal UV radiation is 260 nm because
this wavelength is the most absorbed by DNA.
– mutations  death.
– UV light causes formation of thymine dimers in DNA which
inhibits DNA replication and function.
– DNA damage can be repaired by several repair
mechanisms.
– Longer UV wavelengths (325-400) are also harmful by
inducing the breakdown of tryptophan to toxic products.
97
Radiation damage…
• Visible light is so important but at high intensities it become
harmful by the action of pigments (chlorophyll, flavins,
bacteriochlorophyll and cytochromes) which absorb light and
get excited and act as photosensitizers. These will then
transfer its energy to O2 generating the singlet oxygen (1O2)
• It is powerful oxidizing agent and will quickly destroy a
cell. This is also employed by phagocytic cells to destroy
bacteria after engulfing.
• Many microorganisms avoid the harmful damage of extensive
light by carotenoid pigments.
• This pigment protect many light-exposed
microorganisms from photooxidation by quenching
singlet O2 by converting its energy back to the
unexcited ground state.
98
Growth Limitation by Environmental
Factors
• Leibig’s law of the minimum
– total biomass of organism determined by nutrient present
at lowest concentration. An increase in limiting essential
nutrient such as phosphate will increase the population
until some other nutrient become limiting.
• Shelford’s law of tolerance
– above or below certain environmental limits, a
microorganism will not grow, regardless of the nutrient
supply. Such as extreme temperature or pH, O2 level,
pressure or the presence of other inhibitory substances.
99
Responses to low nutrient levels (oligotrophic
environments)
• The organisms become more competitive in nutrient capture

and use of available resources
• Developing morphological changes to increase surface area and

ability to absorb nutrients. So, some microbes will change their
shape from rod to mini- or ultra micro cells (figure 6-26).
• Developing mechanisms to sequester certain nutrients such as

iron (Fe) making it less available for their competitors
100
Biofilms
• most microbes grow attached to surfaces
(sessile) rather than free floating
(planktonic)
• these attached microbes are members of
complex, slime enclosed communities
called a biofilm
• biofilms are ubiquitous in nature in water
• can be formed on any conditioned surface
101
Figure 7;33
102
Figure 7.34; biofilm heterogeneity
103
Biofilm Formation
• microbes reversibly attach to
conditioned surface and release
polysaccharides, proteins, and DNA
to form the extracellular polymeric
substance (EPS)
• additional polymers are produced as
microbes reproduce and biofilm
matures
104
Biofilms
• a mature biofilm is a complex, dynamic
community of microorganisms
• heterogeneity is differences in metabolic
activity and locations of microbes
• interactions occur among the attached
organisms
– exchanges take place metabolically, DNA
uptake and communication
105
Biofilm Microorganisms
• the EPS and change in attached
organisms’ physiology protects microbes
from harmful agents
– UV light, antibiotics, antimicrobials
• when formed on medical devices, such as
implants, often lead to illness
• sloughing off of organisms can result in
contamination of water phase above the
biofilm such as in a drinking water
system
106
Cell to Cell Communication

Within the Microbial Populations
• bacterial cells in biofilms communicate in
a density-dependent manner called
quorum sensing
• produce small proteins that increase in
concentration as microbes replicate and
convert a microbe to a competent state
– DNA uptake occurs, bacteriocins are
released
107
Quorum Sensing
• acylhomoserine lactone (AHL) is an
autoinducer molecule produced by many gram-
negative organisms to regulate biofilm
formation and increase virulence factors
– diffuses across plasma membrane
– once inside the cell it induces expression of
target genes that regulate a variety of
functions
– many microbes produce effect
108
Figure 7.35; representative cell-cell communication

molecules
109
Quorum Sensing Systems

• processes regulated by quorum
sensing involve host-microbe
interactions; this might lead to;
– symbiosis – Vibrio fischeri and
bioluminescence in squid
– Increase pathogenicity and increased
virulence factor production
– lead to DNA uptake for antibiotic
resistance genes
110
Figure 7.36
111

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Microbial Growth and

Bacterial Cell Cycle

Chromosome Replication and

Z Ring Formation - Role in

Cellular Growth and

Cell Wall Growth and Cell

Cell Wall Growth and Cell

The Growth Curve

Possible Reasons for

Stationary Phase and

• B; it is believed that a fraction of microbes go into

Prolonged Decline in Growth

Measurement of Growth Rate and Generation Time

Figure 7.17; generation time

Direct Counts on Membrane

Viable Counting Methods

Viable Counting Methods

Viable Counting Methods

• Growth in an open system means

• Rate of incoming fresh

• Dilution rate increase means more nutrition so the generation

• This goes up to a level were the dilution rate is too high, so at

• But the biomass will decrease sharply because this high

This is another type of continues growth vessel but with a

This system differs from the chemostat in the following:

– Table 7-4 summarizes the way in which microorganisms

Solutes and Water Activity

Aw of a solution can be measured by sealing the solution in a

Microbes Adapt to Changes in

Extremely Adapted Microbes

– Drastic changes in pH disrupts the plasma membranes of

– Some bacteria synthesize proteins that provide protection

• Very low temperatures solidify the bacterial membranes

• Organisms therefore have characteristic temperature -

• Mesophiles; are microorganisms with growth optimum 20-45 °

Temperature Ranges for

Oxygen and Bacterial Growth

Oxygen and Bacterial Growth

need prefer ignore oxygen is < 2 – 10%

Basis of Different Oxygen

Strict Anaerobic Microbes

Figure 7.29; anaerobic work station

• The organisms become more competitive in nutrient capture

• Developing morphological changes to increase surface area and

• Developing mechanisms to sequester certain nutrients such as

Figure 7.34; biofilm heterogeneity

Cell to Cell Communication

Figure 7.35; representative cell-cell communication

Quorum Sensing Systems

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