BACTERIAL NUTRITION AND GROWTH

By: Mary Ylane S. Lee

• WHAT FACTORS AFFECT THE GROWTH OF
BACTERIA?

A. PHYSICAL OR ENVIRONMENTAL
FACTORS

B. NUTRIENT FACTORS OR REQUIREMENTS

• PHYSICAL REQUIREMENTS or
ENVIRONMENTAL FACTORS
- Temperature
- pH
- Gas (Oxygen and CO2 requirement)
- Osmotic Pressure
TEMPERATURE

Temperature

How does temperature affect

growth rate of bacteria?
CARDINAL TEMPERATURE
• MINIMUM GROWTH TEMPERATURE
• OPTIMUM GROWTH TEMPERATURE
• MAXIMUM GROWTH TEMPERATURE
Sternothermal- species that have small
growth temperature range.

Eurythermal- species which grow over a

wide range of temperature.
Categories based temperatures:

PSYCHROPHILES
- Grow well at 0°C and have an optimum growth
temperature of 15°C or lower and a maximum is
around 20°C .
FACULTATIVE PSYCHROPHILES
-grow at 0°C, have an optima between 20 to 30°C
And maxima of about 35°C
MESOPHILES
- Growth optima is around 20 to 45 °C, a
temperature minimum of 15°C to 20°C and a
maximum of about 45°C or lower
THERMOPHILES
- Can grow at temperatures of 55°C or
higher. Minimum is usually 45°C and have
an optima between 55 and 65°C .
HYPERTHERMOPHILES
- Optima is between 80°C and about
110°C. They usually do not grow well
below 55°C
• Oxygen Requirements: 
How does oxygen affect optimal growth?
-microorganisms fall into several groups with
respect to the effect of oxygen
on their growth and metabolism:
may be Aerobic and Anaerobic
1. Obligate Aerobes- completely dependent on
atmospheric oxygen for growth.
2. Facultative anaerobes- do not require oxygen
for growth but do grow better in its presence.
3. Aerotolerant anaerobes- simply ignore oxygen
and grow equally well, whether it is present or
not.
4. Strict or obligate anaerobes – do not tolerate
oxygen at all and die in its presence.
5. Microaerophiles- damaged by normal
atmospheric level of oxygen and require level
below the range of 2 to 10% for growth.
GROWTH LOCATION BASED ON GASEOUS REQUIREMENT

obligate Facultative Obligate Aerotolerant

Microaerophiles
aerobes anaerobes anaerobes anaerobes
• pH effects:

The majority of organisms live or grow in

habitats between pH 6 and 8 because
strong acids and bases can be
highly damaging to enzymes and other
cellular substances.
Acidophiles
- Optimum between
pH 0 and 5.5
Neutrophiles
- Between 5.5 and 8
Alkalophiles
- Range of 8.5 to 11.5
 Osmotic Pressure/ Water Activity

Osmotic Pressure:
• It is the force with which a solvent moves
from a solution of lower solute concentration
to a solution of higher solute concentration
Hypertonic Hypotonic
• water must be available for metabolism
and growth (80-90% of cell mass)
water activity aw: represents the mole
fraction of the total water molecules that
are available:
• aw = p = vapor pressure of solution
p0 vapor pressure of water w
measured by its relationship to relative
humidity (aw x 100 = RH)
• What is the minimum amount of water
allowing growth?
lowest water activity allowing growth:
most bacteria 0.91
most yeasts 0.88
most molds 0.80
halophilic bacteria 0.75
xerophilic fungi 0.65
osmophilic yeasts 0.60
Most bacteria require an isotonic or
hypotonic environment for optimum
growth.
Osmotolerant- grow over wide ranges
of water activity or osmotic pressure.
Halophiles- require high concentration
of sodium chloride to grow.
•

NUTRITIONAL REQUIREMENTS
• Essential Nutrients: 
• Are any molecular or elemental form of nutrient that is
required by an organism. 

• Two categories of essential nutrients; macro-nutrients

and micro-nutrients.
– Macro-nutrients are needed in larger amounts.
• Used to help with cell structure and the cell's metabolism.
• Examples are proteins, and carbohydrates.
– Micro-nutrients or trace elements are needed in a lot smaller
amount.
– They help enzyme function and help to maintain protein
structure.
– They include elements such as zinc, manganese, and
nickel.
 ELEMENT CELL FUNCTION

C backbone of organic cell components, energy

water, organic components, pH, hydrogen bonds, re-

H
dox
O water, organic components, respiration
N amino acids, nucleotides, coenzymes, ATP
S amino acids, coenzymes, enzymes

P nucleic acids, phospholipids, coenzymes, ATP

Na, K, Ca, Cl, Mg, Trace elements: transport, ionic balance, cofactors
Mn (e- donor/acceptors)
• THE SOURCE OF COMMON ESSENTIAL
NUTRIENTS ARE: CHNOPS

• Carbon
• Hydrogen
• Nitrogen
• Oxygen
• Phosphorous
• Sulfur
• Carbon Content Nutrients:
Carbon content is another way that
nutrients are categorized. Sources of
nutrients is extremely varied and some
microbes will obtain their nutrients entirely
from inorganic sources and others require
a combination of organic and inorganic
sources.
• Organic nutrients:
– Contain at least some combination of
carbon and hydrogen atoms.
– Natural organic molecules are usually products of
livings things.
– Simple to large polymers.
• Inorganic Nutrients:
– An element or simple molecule that contains elements
other than carbon and hydrogen.
– Natural reservoirs are mineral deposits in the crust of
the earth, bodies of water and the atmosphere.
• EX: metals and their salts (magnesium, sulfate,
ferric nitrate, sodium phosphate).
• EX: Gases (oxygen, carbon dioxide) and water
 According to Energy Source

• Energy Source
– Phototroph
• Uses light as an energy source
– Chemotroph
– Uses energy from the oxidation of reduced
chemical compounds
 According to Electron Source

• Electron (Reduction potential) Source

– Organotroph
• Uses reduced organic compounds as a source for
reduction potential
– Lithotroph
• Uses reduced inorganic compounds as a source for
reduction potential
• Carbon Sources
Even though there is a distinction between
the types of carbon cells can absorb as
nutrients. It is important to know that the
majority of carbon involved in the structure
and metabolism of all cells are organic
• Prokaryotes can be divided into various
physiological groups based on how
they derive energy and assimilate
carbon
 According to Carbon Source

• Carbon source
– Autotroph
• Can use CO2 as a sole carbon source
(Carbon fixation)
– Heterotroph
• Requires an organic carbon source; cannot use CO2
as a carbon source
Can also be classified as:

Group Carbon from Energy by

Chemolithotrophs CO2 oxidation of
inorganic
compounds

Photolithotrophs CO2 light

Chemoheterotrophs organic compounds oxidation of

inorganic
compounds

Photoheterotrophs organic compounds light

Representative microorganisms:
• Chemolithotrophs (chemolithotrophic autotroph):
- sulfur oxidizing bacteria
- nitrifying bacteria
• Photolithotrophs (photolithotrophic autotroph)
- algae
- cyanobacteria
- purple and green sulfur bacteria
• Chemoheterotroph (chemoorganotrophic heterotroph)
- nonphotosynthetic bacteria
- fungi
- protozoa
• Photoheterotrophs (photoorganotrophic heterotroph)
- purple and green non-sulfur bacteria
• Nitrogen source
– Organic nitrogen
• Primarily from the catabolism of amino acids
– Oxidized forms of inorganic nitrogen
• Nitrate (NO32-) and nitrite (NO2-)
– Reduced inorganic nitrogen
• Ammonium (NH4+)
– Dissolved nitrogen gas (N2) (Nitrogen fixation)
• Phosphate source
– Organic phosphate
– Inorganic phosphate (H2PO4- and HPO42-)
• Sulfur source
– Organic sulfur
– Oxidized inorganic sulfur
• Sulfate (SO42-)
– Reduced inorganic sulfur
• Sulfide (S2- or H2S)
– Elemental sulfur (So)
Nutrient Transport Processes

• Simple Diffusion
– Movement of substances directly across a
phospholipid bilayer, with no need for a
transport protein
– Movement from high  low concentration
– No energy expenditure (e.g. ATP) from cell
– Small uncharged molecules may be
transported via this process, e.g. H2O, O2, CO2
Nutrient Transport Processes

• Facilitated Diffusion
– Movement of substances across a membrane
with the assistance of a transport protein
– Movement from high  low concentration
– No energy expenditure (e.g. ATP) from cell
– Two mechanisms: Channel & Carrier Proteins
Nutrient Transport Processes

• Active Transport
– Movement of substances across a membrane
with the assistance of a transport protein
– Movement from low  high concentration
– Energy expenditure (e.g. ATP or ion gradients)
from cell
– Active transport pumps are usually carrier
proteins
Nutrient Transport Processes

• Active Transport (cont.)

– Active transport systems in bacteria
• ATP-binding cassette transporters (ABC
transporters): The target binds to a soluble
cassette protein (in periplasm of gram-negative
bacterium, or located bound to outer leaflet of
plasma membrane in gram-positive bacterium).
The target-cassette complex then binds to an
integral membrane ATPase pump that transports
the target across the plasma membrane.
Nutrient Transport Processes

• Active Transport (cont.)

– Active transport systems in bacteria
• Cotransport systems: Transport of one
substance from a low  high concentration as
another substance is simultaneously transported
from high  low.

For example: lactose permease in E. coli:

As hydrogen ions are moved from a high
concentration outside  low concentration inside,
lactose is moved from a low concentration outside
high concentration inside
Nutrient Transport Processes

• Active Transport (cont.)

– Active transport systems in bacteria
• Group translocation system: A molecule is
transported while being chemically modified.

For example:
phosphoenolpyruvate: sugar phosphotransferase
systems (PTS)

PEP + sugar (outside) pyruvate + sugar-

phosphate (inside)
• Bacterial Growth
- Growth is an orderly increase in the
quantityof cellular constituents.
- Increase in cell numbers
- Bacteria grow by binary fission.
BINARY FISSION
PHASES OF BACTERIAL GROWTH
Lag phase - synthesis of new components or
repair
– there is little or no change in the number of
cells, but metabolic activity is high.
– DNA and enzyme synthesis occurs; may last
from 1 hour to several days.


Log phase - reproduction at maximum rate
(shortest generation time)
– the bacteria multiply at the fastest rate
possible under the conditions provided.
Stationary phase - no net increase, balance
between cell division, cell "death",
- there is an equilibrium between cell
division and death.

Death phase- the number of deaths

exceeds the number of new cells formed.
GENERATION TIME
 The time required for a cell to divide or a
population to double is known as the generation
time.
 
• quite short; about 20 –60 minutes under
optimal condition

Most bacteria have a doubling time of 1-3 hours,

although some may be greater than 24 hours.

• 5 –10 hours for most pathogenic bacteria

GENERATION TIME

• . . . is defined as the relationship

between the number of bacteria in a
population at a given time (Nt), the
original number of bacterial cells in the
population (No), and the number of
divisions those bacteria have
undergone during that time (n) can be
expressed by the following equation:
Nt = No x 2n
The bacterial
growth curve:
• Lag phase
• Exponential
phase
• Stationary phase
• Death phase
During the exponential phase, the
number of cells doubles every
generation (1, 2, 4, 8, 16...) and the
number of cells in the culture (Nf) at
any particular time is given by:
Nf = (Ni)2n
 Nf = (Ni)2n
where:
Ni is the initial size of the population
n is the number of generations that have
elapsed.
This equation is usually written in the form
log10 for mathematical convenience:
log10 Nf  = log10 Ni + 0.301n(log10 2 = 0.301)
• From this equation it is possible to
determine n (generations elapsed) by
knowing the numbers of cells at the
beginning and end of a particular
incubation period:
n = (log10Nf - log10Ni) / 0.301
• This is useful because it enables
calculation of the generation time g (the
time between the beginning and end of the
incubation period:
g = t/n
EXAMPLE:

• 100 bacteria present at time 0

if generation time is 2 hours,
what is the cell mass after 8 hours?
Cell Mass= No x 2n
= 100 x 24
= 1,600 bacteria
Sample Problem 1

Bacillus cereus divides every 30 minutes.

You inoculate a culture with exactly 100
bacterial cells. After 3 hours, how many
bacteria are present?
Solution

In 3 hours, B. cereus will divide 6 times.

Therefore, n = 6. 26 = 64 or 2x2x2x2x2x2
100 x 64 = 6,400 cells
 Using the same example, let’s say you
have determined that your sample
contains 6,400 bacterial cells. You know
that it incubated 3 hours. How many
generations have occurred?
• Number of generations = (log cells at
end of incubation ) - (log cells at
beginning of incubation) / 0.301
n = (log10Nf - log10Ni) / 0.301

• Therefore, (log 6400) - (log 100) / 0.301 =

(3.81 - 2) / 0.301 = 6 generations
• To calculate the generation time for a
population: 60 min x hours / number of
generations
g = t/n

In this example:
• 60 min x 3 hours / 6 generations = 30
minutes per generation
• Practice Problems
• 1. You perform a serial dilution and determine that the original
number of cells in your sample was 12, 000. How many bacteria
will be present in 12 hours if the generation time is 15 minutes
(assume unlimited food and clean environment)?

• 2. You determine that a coconut cream pie has 3 million (3 x

106) Staph. aureus cells in it. You estimate that the food
preparer did not wash his hands and probably inoculated the
cream with 500 Staph. aureus. He also forgot to refrigerate it. If
the pie was made 6 hours ago, how many generations have
occurred? How long is each generation?
•
• 3. Using the generation time from problem 2, how many bacteria
would be present after 8 hours at room temperature?

• 4. Let’s say that flesh eating Strep. pyogenes divides every 10

minutes at body temperature. You fall down and scrape your
knee and get infected with 5 Strep. pyogenes cells. After 4
hours, without medical intervention, how many bacteria will be
ravaging your body?