Sasha Anne L.

Valdez
OUTLINE FOR MODULE 4 Part 2
I. Module Objectives

II. Factors Affecting Microbial Growth

III. Physical (Environmental) Factors Affceting Microbial

Growth
1. pH
2. temperature
3. oxygen
4. moisture
5. osmotic pressure
6. hydrostatic pressure
7. radiation

IV. Microbial Reproduction, Binary Fission, and Bacterial

Growth Phases
Module 4: Microbial Growth Requirements Part 2
Module Objectives:

1. Identify environmental/physical factors that could

affect microbial growth.
2. Classify microorganisms based on environmental
conditions they can thrive in.
3. Predict growth behavior of microbes based on growth
conditions.
4. Recognize the significance of knowledge on
environmental conditions and nature of
microorganisms in relation to successful laboratory
microbial cultivation.
5. Understand bacterial mode of reproduction.
6. Define and describe each bacterial growth phase
7. Use mathematical expression to calculate for
generation time and solve bacterial growth problems.
 microorganisms can be found in nearly
every environment on earth

 small and easily dispersed

 occupy little space
 need only small quantities of nutrients
 diverse in their nutritional requirements
 great capacity for adapting to environmental changes

kinds of organisms and rate of organism growth

can be influenced by many factors:
physical and biochemical (nutritional)
Factors Affecting Microbial Growth

• biochemical
factors (nutrition) • physical factors
– macronutrients – pH
– micronutrients – temperature
– vitamins – oxygen concentration
– moisture
– osmotic pressure
– hydrostatic pressure
– radiation
• generation time
 pH

• pH = hydrogen ion concentration

• neutrality is pH 7 (optimum pH for most
bacteria)
• pH values less than 7 – acidic
• pH values greater than 7 - basic
• optimum pH for most bacteria is near
neutrality (pH 7)
• cytoplasm of most bacteria is pH 7
Optimum pH for bacterial growth
 pH

• each organism has a pH range within

which growth is possible and usually has
a well-defined pH optimum

Classification pH range
Acidophiles Below pH 5.4
Neutrophiles pH 5.4 - 8.5
Alkaliphiles pH 7.5 – 11.5
https://courses.lumenlearning.com/microbiology/chapter/the-
effects-of-ph-on-microbial-growth/
 pH
• acidophiles
– acid-loving organisms
– can be found in acidic lakes, gastrointestinal tract
– most fungi (acid-tolerant; optimum temperature 5 or below)
– some algae, bacteria, and several Archaea
• Lactobacillus (pH 6)
• Helicobacter pylori (pH 2 or less)
• Acidihiobacillus (sulfur-oxidizing bacteria) (pH <4)
• red alga Cyanidium caldarium, green alga Dunaliella acidophila (pH <1)
• fungi: Aconitum cylatium, Cephalosporium sp., Trichosporon cerebriae
(near pH 0)
• archaea: Sulfolobus and Thermoplasma, Picrophilus (negative pH values)
– high H+ concentration is required to maintain cell membrane
stability
 Lactobacillus

https://www.medicalnewstoday.com/ar
https://www.sciencephoto.com/media/9381
https://yakult.com.mt/i ticles/323169
19/view/lactobacillus-bulgaricus-yogurt- ngredients/
bacterium-sem

https://njaes.rutgers.edu/fs
https://medium.com/@Klusch_Nutri/stra
547/
https://www.shutterstock.com/image- wberries-nutritional-content-spoilage-
photo/mold-fungus-citrus-fruits-isolated-on- Fungi and-preservation-methods-
86448391 a4b2e24b48a0
 https://www.medicalnewstoday.com/articles/309046 https://www.quora.com/Is-a-volcanic-acidic-lake-really-
dangerous

Helicobacter pylori in stomach

(pH ≈ 2) may cause chronic a volcanic acidic lake (top picture) and an
gastritis and gastric ulcers acidic hot spring (bottom picture): where
acidophiles would thrive

https://www.2minutemedicine.com/patient-basics-
peptic-ulcer/ https://www.huffpost.com/entry/yellowstone-hot-
springs-death-hot-pot_n_582cf703e4b058ce7aa9258f
http://article.sapub.org/10.5923.j.microbiology.201
20204.03.html https://alchetron.com/Thermoplasma-volcanium

Thermoplasma
Acidithiobacillus thiooxidans
Sulfolobus acidocaldarius

Picrophilus

https://microbewiki.ken
yon.edu/index.php/Picr
http://web.pdx.edu/~ksted
ophilus
man/research.html
 pH

 neutrophiles
 most human disease-causing bacteria
(human blood and tissues pH = 7.2 – 7.4)
 protozoans and most bacteria (pH 6.5-7.5)
E. coli bacteria in gut

https://www.sciencephoto.com/media/1
2516/view/escherichia-coli-bacteria-in-
https://www.news-medical.net/health/Sepsis- the-gut
Diagnosis.aspx
 https://www.dreamstime.com/balantidium-
coli-protozoan-large-intestine-d-
illustration-ciliated-intestinal-parasite-
causes-balantidiasis-image128971571

Salmonella bacteria on tissue surface

Balantidium coli (protozoan) in

human large intestines

https://focusedcollection.com/160559728/stock-
photo-salmonella-bacteria-on-tissue-surface.html

Staphylococcus skin infection

https://www.healthdirect.gov.au/staph-skin-infection
 pH
 alkaliphiles
 base-loving organisms
 live in soda lakes, high carbonate soils
i.e. Bacillus, Vibrio cholerae (pH 9), Alcaligenes
faecalis (>pH 9), Agrobacterium (pH 12)
 some produce hydrolytic enzymes (proteases and
lipases)
Soda lake (left) and
alkaline hot spring
(right): where
alkaliphiles may
thrive

https://microbewiki.kenyon.edu/index.php/Alk
aline_hot_springs
https://www.ripleys.com/weird-news/soda-lake/
 Vibrio cholerae

Agrobacterium

https://www.shutterstock.com/search/vibrio+cholerae

Alcaligenes faecalis

https://www.sciencephoto.com/media/13176/view/a
grobacterium-tumefaciens

https://www.sciencephoto.com/media/1211
2/view/alcaligenes-faecalis
http://www2.nau.edu/~fpm/bio205/chapter6.html
 Tolerating Acidity/Alkalinity
• impervious cell walls protect cell membrane
and internal contents of cell
• acid by-product inhibits microbial growth
• secrete biofilm to slow down diffusion of
molecules into cell
• secrete buffer molecules or enzymes (i.e. H.
pylori and urease)
• cells maintain a constant internal pH (near
neutral)
• pumping of H+ out of the cell at a high rate or
exchanging sodium ions for external protons
 Temperature
• one of the most, if not the most, important
environmental factors affecting growth and
survival of microorganisms

https://www.sciencelearn.org.nz/image_maps/60-temperature-the-
hot-and-the-cold
 Temperature
three critical temperatures (affecting enzyme
function) or cardinal temperatures:
minimum growth temperature – lowest temperature
at which cells can divide (a)
• membranes solidify; slow transport process thus
growth could not occur
maximum growth temperature – highest temperature
at which cells can divide (b)
• protein denaturation, collapse of cell membrane,
cell lysis
optimum growth temperature – temperature at
which cells divide most rapidly (c)
• enzymatic reaction occurring at maximal possible
rate
 b

cardinal temperatures are generally

characteristic of each type of
microorganism and differ widely from
different types of microorganisms
https://courses.lumenlearning.com/microbiology/chapter/temperature-and-microbial-growth/
Microorganism Temperature Optimum
Range Temperature
Psychrophiles: <0 to 20C 15 C
Mesophiles 10 to 48C 37 C
Thermophiles 40 to 72C 60 C
Hyperthermophiles 65 to 110C 80 C

27
Psychrophiles

 “cold-loving organisms”
 grow best at -10˚ to 20˚C
 obligate psychrophiles (Sporosarcina
globispora, <20 ˚C)
 facultative psychrophiles (Xanthomonas
pharmicola, above or below 20 ˚C)
 snow alga: Chlamydomonas nivalis
 Listeria monocytogenes (can multiply at -4.4
˚C)
 live mostly in cold water and soil (Arctic and
Antarctic regions) and can cause spoilage of
refrigerated food
https://www.siliconrepublic.com/innovation/ https://www.flickr.com/photos/18745440@N
polar-ice-living-bacteria-alien-life 00/2579153321/lightbox/

Chlamydomonas nivalis
Psychrophiles may exist in (snow alga)
polar ice

Psychrophiles may thrive inside

refrigerators

https://www.express.co.uk/news/science/896871/Bacteria-alien-
polar-ice-planets-Titan-microbes-climate-change-CO2
 FACT SHEET OF Listeria monocytogenes

https://saniprofessional.com/listeria-monocytogenes-fact-sheet/
Mesophiles

➢ most bacteria including pathogens

➢ most common group of microorganisms
➢ 25 ˚to 40˚C
➢ found in warm-blooded animals
➢ thermoduric microorganisms (Bacillus,
Micrococcus, Lactococci, Corynebacterium)
• can withstand short periods of exposure to high
temperatures; can cause food spoilage
Thermophiles Survive pasteurization
G. stearothermophilus – autoclave
validation (1/month); biological indicator for
sterilization
➢ “heat-loving organisms”
➢ 40 ˚ to 72˚C
➢ compost heaps, hot springs
➢ contaminants in dairy products
➢ can be categorized into:
• obligate thermophiles
– temperatures above 37 ˚C
– Geobacillus stearothermophilus (65-75 ˚C)
• facultative thermophiles (moderate thermophiles)
– can grow both above and below 37 ˚C
– thermophilic sulfur bacteria in runoff troughs of geysers
– Bacillus coagulans (35-50 ˚C), B. licheniformis,
Anoxybacillus spp., Paenibacillus spp., Thermoanaerobacter
spp. and Clostridium thermobutyricum /thermopalmarium

https://www.sciencephoto.com/media/709153/view/thermophile-
bacteria
https://girlmeetsbiochemistr
y.wordpress.com/2013/04/1
4/thermophiles-
thermophiles-organisms/
Scanning electron micrograph (SEM) of
thermophilic bacteria (blue). Collected from
140 degree F water in the springs of
Yellowstone National Park, Wyoming USA. This
type of bacteria is adapted to thrive at high
water temperatures and is currently the focus
of biological researchers. Bacteria that can live
in these extreme conditions are called
thermophiles or extremophiles. Typically, the
hotter the temperature the smaller the
bacteria. Shot at 6,580X magnification with a
field of view of 20 um.
 Hyperthermophiles
➢ extreme heat-loving organisms
➢ 65 to 110 ˚C
➢ boiling hot springs, geysers, hot-water vents
• archaeobacteria (deep-sea vents, 115 ˚C)
• Pyrolobus fumarii (“firelobe of the chimney”) – (113 ˚C)
• Thermus aquaticus

Alicyclobacillus acidocaldarius –
geysers in Yellowstone National Park
Alicyclobacillus acidoterrestris -
warm soil hydrothermal vent is a
fissure on the seafloor from which
geothermally heated water issues.
Hydrothermal vents are commonly
found near volcanically active places,
areas where tectonic plates are
moving apart at spreading centers,
ocean basins, and hotspots
https://kaiserscience.wordpress.com/biology-
the-living-environment/cells/hyperthermophiles/
 Temperature
 PSYCHROPHILES: cold-loving
 MESOPHILES: moderate temperature-loving
 THERMOPHILES: heat-loving

Each has a minimum, optimum, and maximum growth

temperature

 above maximum and below minimum temperatures

 growth does not occur
 optimum temperature
 at which enzymes function
Fig. 7.8
 microorganisms’ growth properties are due to changes in
enzyme activity˚
 enzyme activity generally doubles for every 10 ˚C rise in
temperature
 Temperature
 important not only in providing conditions for
growth but also in preventing such growth
 refrigeration (4 ˚C) – reduces growth of
psychrophiles; prevents growth of other
microorganisms (i.e. refrigerator)
 long-time storage - -30 ˚C (i.e. ultra-low freezer)
 high temperatures - prevent bacterial growth (i.e.
pressure cooker)
Bacteria can survive extremes of cold than extremes of heat

https://nanbei-china.en.made-in-
china.com/product/yXSJlCbYfIUh/China-
https://www.pinterest.ph/pin/658 https://fungi.com/collections/cultivation- 668-Liter-86-Degree-Ultra-Low-
018195533024044/ tools/pressure-cookers-and-sterilizers Temperature-Freezer-Refrigerator.html
39
 Tolerating Low and High
Temperatures
 psychrophiles
 unsaturated (polyunsaturated) fatty acids in phospholipids
 with enzymes functional at low temperatures
 active transport occurs well at low temperatures

 thermophiles
 saturated fatty acids in phospholipids
 heat-stable proteins and enzymes

 hyperthermophiles
 no fatty acids in their membrane (phytane)
 lipid monolayer
What are
psychrotrophs or
psychrotolerant
microbes?
 Psychrotolerant
organisms are like
mesophiles (growing at
20-40C), but are able to
tolerate lower
temperatures albeit with
slower growth rate

https://www.differencebet
ween.com/difference-
between-psychrophiles-
and-psychrotrophs/
obligate vs. facultative

• obligate – MUST
• facultative – ABLE to adjust
 Oxygen
 bacteria can be divided into:
 aerobes – require oxygen to grow
 anaerobes – do not require oxygen to grow

 microorganisms can be classified as :

1. obligate aerobes
 must have free oxygen for aerobic respiration
 Pseudomonas
2. obligate anaerobes
 does not require/use oxygen for metabolism
 Bacteroides, Clostridium methanogens, Thiomargarita
namibiensis
 organisms can be found in muds, sediments of lakes, rivers,
oceans, marshes, water-logged soils, canned foods, intestinal
tracts, sewage treatment systems, anoxic environments
O2 is necessary for aerobic cellular
respiration; oxygen oxidize substrates to
produce energy
 Capnophiles – Neisseria

Oxygen and Streptococcus

Capnophiles are

microorganisms can be classified as :

microorganisms that
• thrive in the presence of
high concentrations of
3. microaerophiles carbon dioxide

• grow best in the presence of small amount of free oxygen

• Campylobacter (also a capnophile: organism that requires
high carbon dioxide concentration)
• Treponema pallidum
4. facultative anaerobes
• ordinarily carries aerobic metabolism when oxygen is
present but shifts to anaerobic metabolism when oxygen is
absent
• Staphylococcus and E. coli
• have complex enzyme systems
5. aerotolerant anaerobes
• can survive in the presence of oxygen but do not use it in
their metabolism
• Lactobacillus (captures energy by fermentation)
Why is oxygen toxic to some
microorganisms?
Toxic oxygen
Singlet oxygen, superoxide anion, hydrogen peroxide,
hydrogen radical → damage to cellular components
Aerobes and facultative aerobes have superoxide dismutase
and catalase
Few aerobes lack catalase
Obligate anaerobes lack/low level of SD
Aerotolerant – no SD but has protein free Mn2+, Fe2+, or
Cu2+, Zn2+ (dismutation of superoxide anions)
 Group Relationship to Types of Example Habitat
O2 metabolism
Aerobes
Obligate Required Aerobic Micrococcus Skin, dust
respiration
luteus
Facultative Not required, but Aerobic, Escherichia Mammalian
grows better with anaerobic, large
fermentation
coli
O2 intestine
Microaerophilic Required but at Anaerobic Spirillum Lake water
levels lower than respiration
volutans
atmospheric O2
Anaerobes
Aerotolerant Not required, Fermentation Streptococc Upper
growth not better us pyogenes respiratory
when O2 present tract

Obligate Harmful or lethal Fermentation or Methanobacterium Sewage

anaerobic formicicum, sludge
respiration Clostridium digestors,
anoxic lake
sediments

Anaerobes – lack respiratory system

Oxygen-related growth zones in a
standing test tube
 Bacterial cell distribution in thiogylcolate tubes

In tube A, all the growth is seen at the top of the tube. The bacteria
are obligate (strict) aerobes that cannot grow without an abundant supply of
oxygen.

Tube B looks like the opposite of tube A. Bacteria grow at the bottom of tube
B. Those are obligate anaerobes, which are killed by oxygen.
https://courses.lumenlearning.com/microbiology/chapter/oxygen-requirements-for-microbial-growth/
Tube C shows heavy growth at the top of the tube and growth throughout the
tube, a typical result with facultative anaerobes. Facultative anaerobes are
organisms that thrive in the presence of oxygen but also grow in its absence
by relying on fermentation or anaerobic respiration, if there is a suitable
electron acceptor other than oxygen and the organism is able to perform
anaerobic respiration.

The aerotolerant anaerobes in tube D are indifferent to the presence of

oxygen. They do not use oxygen because they usually have a fermentative
metabolism, but they are not harmed by the presence of oxygen as obligate
anaerobes are.
https://courses.lumenlearning.com/microbiology/chapter/oxygen-requirements-for-microbial-growth/
Tube E on the right shows a “Goldilocks” culture. The oxygen level has to be
just right for growth, not too much and not too little.

These microaerophiles are bacteria that require a minimum level of oxygen

for growth, about 1%–10%, well below the 21% found in the atmosphere.

https://courses.lumenlearning.com/microbiology/chapter/oxygen-requirements-for-microbial-growth/
 Anaerobic Culture Methods
• Use reducing media, containing chemicals (e.g.:
thioglycolate) that combine with O2

• Use anaerobic jar (GasPak)

• Novel method in clinical labs:

Add oxyrase to growth media
 OxyPlate (no need for anaerobic jar)

• Work in a glove box

• Use candle jars

 https://www.nichwell.com/a-1200u-separated-
inert-vacuum-controlled-atmospheres-glovebox/

Glove box

https://www.slideshare.net/SantoshYadav225/method-of-
anaerobiasis-and-anaerobic-culture
https://www.bd.com/en-
us/offerings/capabilities/microbiology-
solutions/environmental-systems/bd-
gaspak-systems-and-accessories/bd-bbl-
gaspak-jars
https://www.slideshare.net/SantoshYadav225/method-of-
anaerobiasis-and-anaerobic-culture
https://www.jfmed.uniba.sk/fileadmin/jlf/Pracoviska/ustav-
mikrobiologie-a-imunologie/ANAEROBIC_BACTERIA.pdf https://basicmedicalkey.com/neisseria-and-
moraxella-catarrhalis/

Candle jar
https://www.ramauniversity.
ac.in/online-study-
material/pharmacy/bpharm
a/iiisemester/pharmaceutic
https://2018.igem.org/Team:Montpellier/Tool
almicrobiology/lecture-
box
11.pdf
 OxyPlates™ create anaerobic
environments in a unique way ... from the
plate up. The agar plate is actively
reduced by Oxyrase®. Anaerobic
microbes put on that surface are
protected from oxygen until the
environment inside the OxyDish™ is
supportive of their growth.
https://www.bioquote.com/product/oxypla
tes-schaedler-blood-agar/ OxyPlates
OxyPlates™ offer many advantages over traditional anaerobic methods.
Streak the OxyPlate™ and incubate it in a standard incubator aerobically.
OxyPlates™ can be opened and closed numerous times regenerating
anaerobic conditions.

OxyPlates™ can be used for subbing anaerobe isolates from

primary plates in all laboratories, regardless of the method used
or the source of primary media.
 Moisture or Water
• Water – essential ingredient of bacterial
protoplasm.

• water is needed by actively metabolizing

cells

• Effect of drying varies:

– Treponema pallidum – highly sensitive
– Staphylococcus sp. – can stand for months
– endospore-former bacteria and xerophiles –
resistant to desiccation
 Osmotic Pressure
 osmotic pressure – minimum pressure needed to be applied
to a solution to prevent the flow of water across a semi-
permeable membrane

 most bacteria require an isotonic environment or a

hypotonic environment for optimum growth

 osmotic pressure outside > osmotic pressure inside the cell

 hyperosmotic environment → plasmolysis

 too high osmotic pressure outside cell → water loss →

inhibits growth or kill bacterial cells
 Osmotic Pressure
 application: use of salt or sugar as preservative
 salting of fish
 sugaring of fruits
 brining of vegetables
 jams, marmalades, preserves, and pickles

 osmotolerant - organisms that can grow at relatively

high salt concentration (up to 10%)

 halophiles – salt-loving organisms; require relatively

high salt concentrations for growth (i.e. archea require
NaCl concentrations of 20 % or higher)

➢ have transport systems to regulate movement of

substances
https://preppers101blog.wordpress.com/2016/01/
https://web.facebook.com/mypicklesandja
16/how-to-use-salt-and-smoke-to-cure-meat-and-
ms/?_rdc=1&_rdr
fish/

http://www.tasteofsouthern.com/how-to-freeze- https://kimchimari.com/my-first-kimjang-at-home-tips-
strawberries-in-sugar/ on-brining-cabbage-for-kimchi/
https://slideplayer.com/slide/7469943/

64
Halophiles
 require moderate to large quantities of salt
 membrane transport system actively transport
sodium ions out of the cell and concentrate potassium
ions in
 typically found in the ocean (optimum: 3.5% salt
concentration)
 found in exceptionally salty bodies of water (Dead
Sea, brine vats)

 Classification
 low halophiles – 1-6% NaCl
 mild or moderate halophiles – 6-15% NaCl
 extreme halophiles – 15-30% NaCl
 https://utex.org/products/utex-lb-0200

Dunaliela salina thriving in the

Dead Sea

Dead Sea fungus

https://jgi.doe.gov/salt-needed- https://www.deadsea.com/articles-tips/interesting-
tolerance-lessons-from-a-dead-sea- facts/why-is-the-dead-sea-called-the-dead-sea/
 Hydrostatic Pressure
• pressure exerted by standing water, in
proportion to its depth
• doubles with every 10 meter increase in
depth
• i.e. 50-m deep lake – 32x atmospheric
pressure
• piezophiles (barophiles) – bacteria that
live at high pressures
– membranes and enzymes (3-D
configuration) require high pressure to
function properly
https://microbewiki.kenyon.edu/index.php/Piezophiles
 Radiation/Radiant Energy
• visible light is the source of energy for
photosynthesis (photosynthetic microorganisms)
Cyanobacteria
(photosynthetic
bacteria)

Volvox
researchgate.net/figure/Cyanobacterial-strains-that-produce-
(photosynthetic
BMAA-representing-different-morphological- green alga)
sections_fig1_7927724

http://beachapedia.org/Cyanobacteria
https://ibiologia.com/algae/
 Radiation/Radiant Energy
• ionizing radiation (gamma rays and UV light)
can cause mutations in DNA and can even kill
microorganisms

• some organisms have pigments that screen

radiation and help to prevent DNA damage

• other organisms have enzyme systems that

can repair certain kinds of DNA damage
https://www.mdpi.com/1422-0067/18/12/2749
 Radiation/Radiant Energy
• some organisms have pigments that screen radiation
and help to prevent DNA damage

• other organisms have enzyme systems that can

repair certain kinds of DNA damage

Halophilic archaea with DNA

repair machinery and
carotenoid pigmentation as
photoprotective strategies
UV resistance strategies of halophilic
https://www.ncbi.
nlm.nih.gov/pmc/ archaea in Great Salt Lake
articles/PMC562
6843/
https://slideplayer.com/slide/12424643/
Microbial Reproduction

How do bacteria reproduce?

• most bacteria reproduce by binary fission

https://www.thoughtco.com/binary-fission-vs-mitosis-similarities-and-differences-4170307
https://nanopdf.com/download/growth-amp-binary-fission-in-bacteria-dr-baha-h-al-
amiedi_pdf
 Binary Fission

1. Replication of chromosome

2. Cell grow in size (double)

3. Septum formation

4. Completion of septum with

formation of distinct walls

5. Cell separation

generation – interval for the formation of two cells from

one cell
 Generation Time
interval of time between two cell divisions

the time required for a bacterium to give rise to 2

daughter cells under optimum conditions

population doubling time

exponential growth

time is variable and dependent on many factors

https://www.pinterest.ph/pin/350506783479826314/
Rate of Growth of a Microbial Culture:
Arithmetic vs. Logarithmic Scale
Factors Affecting Microbial Growth

• biochemical
factors (nutrition) • physical factors
– macronutrients – pH
– micronutrients – temperature
– vitamins – oxygen concentration
– moisture
– osmotic pressure
– hydrostatic pressure
– radiation
• generation time
 Generation Time

◼ Escherichia coli – 20 mins

◼ Staphylococcus aureus- 27-30 mins

◼ Mycobacterium tuberculosis - 792-932 mins

◼ Treponema pallidum -1980 mins

 Calculating Generation Times
• N = N02n
• N = final cell number
• N = initial cell number
0

• n = number of generations that have occurred

during the period of exponential growth

• g = t/n
• g = generation time
• t = hours or minutes of exponential growth
Calculating Generation Times
• n= log N – log N0 = log N – log N0
log2 0.301
=3.3 (log N – log N )
o

• k = ln 2/g = 0.693/g
k = number of generations that occur per unit time in
an exponentially growing culture
g = generation time
 Bacterial Population Growth Cycle
• bacterium in a suitable medium, incubated, growth
follows a definite course

• 4 phases of bacterial growth curve:

– Lag
– Log or Exponential
– Stationary
– Decline/Death phase

Do not apply to individual cells,

only to population of cells
https://www.researchgate.net/figure/Fig-2-Bacterial-growth-
curve-indicating-various-phases-Stationary-phase-
cultures_fig1_311664511
https://cellgrowth.weebly.com/growth-rate-and-conditions.html
 Phases of Growth Curve
◼ 1. Lag phase – No significant or immediate increase
in cell numbers but there may be an increase in the
size of the cell.
◼ adaptation phase
◼ cell is metabolically active (synthesize enzymes
and new components; utilizes various molecules
from the medium; produce large quantities of
energy)
◼ length –dependent in the characteristics of the
bacterial species and conditions in the media
(“old and new”, “rich and poor”)
◼ period when microorganisms are introduced into
fresh culture medium
 Phases of Growth Curve
◼ 2. Log (Logarithmic) or Exponential phase – cells
start dividing and their number increases
exponentially
◼ organisms divide at their most rapid rate
◼ the population of organism doubles in each
generation time
◼ cells in their healthiest state
◼ influenced by environmental conditions and
genetic characteristics of organisms
◼ bacteria are more susceptible to antibiotics
◼ population is most uniform in terms of chemical,
metabolic, and physiologic properties
 Phases of Growth Curve
◼ 2. Log (logarithmic) or Exponential phase
◼ production of primary metabolites
(ethanol, lactic acid, amino acids) happen
◼ limited period of time
◼ as number of organisms increases,
nutrients are used up, metabolic wastes
accumulate, living space become limited,
aerobes suffer from oxygen depletion
◼ limiting factors affect rate of ATP
production (leveling off of growth)
 Phases of Growth Curve
▪ 3. Stationary phase – cell division decreases
due to depletion of nutrients & accumulation of
toxic products; inadequate oxygen supply; pH
change
▪ growth rate tapers off
▪ equilibrium exists between dying cells and
the newly formed cells (new cells produced
at the same rate as old cells die)
▪ cell functions continue
▪ usually production of secondary
metabolites (small, bioactive molecules)
occur
 Phases of Growth Curve
◼ 4. Decline (Death) Phase - population
decreases due to the death of cells
◼ conditions in the medium become less and
less supportive of cell division
◼ number of live cells decreases at a
logarithmic rate (but slower)
◼ cells undergo lysis or involution (assume a
variety of unusual shapes)
◼ more spores than vegetative cells survive
 Morphological & Physiological
Alterations During Growth
• Lag phase – maximum cell size towards the end of
lag phase.

• Log phase – smaller cells, stain uniformly

• Stationary phase – irregular staining, sporulation

and production of exotoxins

• Phase of Decline –involution forms (with aging)

https://courses.lumenlearning.com/microbiology/chapter/how-microbes-grow/
Why is the growth curve for
total count different to that
of the viable count curve?