Microbial growth and Biokinetics

Classification Of Microorganisms
Classification by Energy and Carbon Source:
The relationship between the source of carbon and the source
of energy for the microorganism is important. Carbon is the
basic building block for cell synthesis.
A source of energy must be obtained from outside the cell to
enable synthesis to proceed.
The goal in wastewater treatment is to convert both the carbon
and the energy in the wastewater into the cells of
microorganisms, which can be removed from the water by
settling or filtration.
Therefore, the processes are designed and operated to
encourage the growth of organisms that use organic material
for both their carbon and energy source.
 If microorganisms use organic material as a supply of
carbon, they are called heterotrophic.
Autotrophs require only CO 2 to supply their carbon
needs. Organisms that rely only on light for energy
are called phototrophs.
Chemotrophs extract energy from organic or
inorganic oxidation/ reduction reactions.
Organotrophs use organic materials, while lithotrophs
oxidize inorganic compounds (Rittmann and McCarty,
2001).
 Classification by Oxygen Relationship:
Bacteria also are classified by their ability or inability to utilize oxygen
in oxidation-reduction reactions.
Aerobes are microorganisms that must have oxygen.
When wastewater contains oxygen and can support aerobes, it is
called aerobic.
Anaerobes are microorganisms that cannot survive in the presence of
oxygen.
Wastewater that is devoid of oxygen is called anaerobic.
Facultative anaerobes can use oxygen in oxidation/reduction
reactions and, under certain conditions, they can also grow in the
absence of oxygen.
Under anoxic conditions, a group of facultative anaerobes called
denitrifies utilize nitrites (NO2-) and nitrates (NO3-) instead of oxygen.
Nitrate nitrogen is converted to nitrogen gas in the absence of oxygen.
This process is called anoxic denitrification.
 Classification by Temperature:
Each species of bacteria reproduces best within a limited range of
temperatures.
Four temperature ranges are used to classify bacteria. Those that grow best at
temperatures below 20 oC are called psychrophiles.
Mesophiles grow best at temperatures between 25 oC and 40 oC. Between 45
oC and 60 oC, the thermophiles grow best.

From about 60 C to near boiling, hyperthermophiles grow best.

The growth range of facultative thermophiles extends from the thermophilic
range into the mesophilic range.
Growth is not limited to these ranges.
Bacteria will grow at slower rates over a larger range of temperatures and will
survive at a very large range of temperatures.
If frozen rapidly, they and many other microorganisms can be stored for years
with no significant death rate.
Once the optimum temperature range is exceeded, growth rate drops off
rapidly due to the denaturation of key proteins.
Some Microbes of Interest in Wastewater
Treatment
Bacteria.
The highest population of microorganisms in a
wastewater treatment plant will belong to the bacteria.
They are single celled organisms that use soluble
food.
Conditions in the treatment plant are adjusted so that
chemoheterotrophs predominate.
No particular species is selected as the best.
 Fungi
Fungi are multicellular, nonphotosynthetic,
heterotrophic organisms.
Fungi are obligate aerobes that reproduce by a
variety of methods including fission, budding, and
spore formation.
Their cells require only half as much nitrogen as
bacteria so that in a nitrogen deficient wastewater,
they predominate over the bacteria (McKinney, 1962).
 Algae.
This group of microorganisms are photoautotrophs and
may be either unicellular or multicellular.
Because of the chlorophyll contained in most species,
they produce oxygen through photosynthesis.
In the presence of sunlight, the photosynthetic
production of oxygen is greater than the amount used
in respiration.
At night they use up oxygen in respiration. If the
daylight hours exceed the night hours by a reasonable
amount, there is a net production of oxygen.
 Algae are of benefit in stabilization lagoons for
wastewater treatment when they supply oxygen in
excess of respiration.
Other than production of oxygen, they do not
contribute to the stabilization of waste because they
use carbon dioxide or bicarbonates as a source of
carbon rather than organic carbon.
They are a liability when they leave in the lagoon
effluent because they contribute to the total
suspended particulate concentration and may cause
discharge limits to be exceeded
 Protozoa:
Protozoa are single-celled organisms that can
reproduce by binary fission (dividing in two).
Most are aerobic chemoheterotrophs, and they often
consume bacteria.
They are desirable in wastewater effluent because
they act as polishers in consuming the bacteria.
 Rotifers and Crustaceans.
Both rotifers and crustaceans are animalsaerobic,
multicellular chemoheterotrophs. The rotifer derives its name
from the apparent rotating motion of two sets of cilia on its
head.
The cilia provide mobility and a mechanism for catching food.
Rotifers consume bacteria and small particles of organic matter.
Crustaceans, a group that includes shrimp, lobsters, and
barnacles, are characterized by their shell structure.
They are a source of food for fish and are not found in
wastewater treatment systems to any extent except in under-
loaded lagoons. Their presence is indicative of a high level of
dissolved oxygen and a very low level of organic matter.
Population Dynamics in Mixed culture
A mixture of species compete and survive within the limits
set by the environment.
Population dynamics is the term used to describe the time
varying success of the various species in competition.
It is expressed quantitatively in terms of relative mass of
microorganisms.
If each individual organism of species A has, on the
average, twice the mass at maturity as each individual
organism of species B, and both compete equally, one
would expect that both would have the same total biomass,
but that there would be twice as many of species B as there
would be of A.
 The prime factor governing the dynamics of the various
microbial populations in sewage is the competition for
substrate (organic matter in the wastewater that is
colloquially called food ).
The relative success of a pair of species competing for
the same substrate is a function of the ability of the
species to metabolize the substrate.
The more successful species will be the one that
metabolizes the substrate more completely.
In so doing, it will obtain more energy for synthesis and
consequently will achieve a greater mass.
 Because of their relatively smaller size and, thus, larger surface
area per unit mass, which allows a more rapid uptake of substrate.
When the supply of soluble organic substrate becomes exhausted,
the bacterial population is less successful in reproduction and the
predator populations increase.
In a closed system with an initial inoculum of mixed
microorganisms and substrate, the populations will cycle as the
bacteria give way to higher level organisms that in turn die for lack
of substrate and are then decomposed by a different set of
bacteria.
In an open system, such as a wastewater treatment plant or a
river, with a continuous inflow of new substrate, the predominant
populations will change through the length of the plant.
 For the large numbers and mixed cultures of
microorganisms found in waste treatment systems, it is
convenient to measure biomass rather than numbers of
organisms.
Frequently, this is done by measuring the suspended
solids or volatile suspended solids (VSS).
This condition is known as dynamic equilibrium.
It is a highly sensitive state, and changes in influent
characteristics must be regulated closely to maintain the
proper balance of the various populations.
Microbial Growth
 A bacteria is introduced into the reactor setup consisting of organic
substrate.
Initially nothing appears to happen.
The bacteria must adjust to their new environment and begin to
synthesize new cells.
On a plot of bacterial growth versus time, this phase of growth is
called the lag phase.
In the start-up of new plants, the lag phase may show a decrease
in population.
At the end of the lag phase the bacteria begin to divide.
Because all of the organisms do not divide at the same time, there
is a gradual increase in population.
This phase is labeled accelerated growth on the growth plot.
 At the end of the accelerated growth phase, the
population of organisms is large enough and the
differences in generation time are small enough that the
cells appear to divide at a regular rate.
Because reproduction is by binary fission (each cell
divides producing two new cells), the increase in
population follows in geometric progression: 1 2 4
8 16 32, and so forth.
This phase of growth is called the log growth or
exponential growth phase.
 The log growth phase tapers off as the substrate becomes
exhausted or as toxic byproducts build up.
Thus, at some point the population becomes constant either
as a result of cessation of fission or a balance in death and
reproduction rates.
This is depicted by the stationary phase on the growth
curve.
Following the stationary phase, the bacteria begin to die
faster than they reproduce.
This death phase is due to a variety of causes that are
basically an extension of those that lead to the stationary
phase.
The Monod Equation
In the log-growth phase, the rate expression for biomass increase
is

Because of the difficulty of direct measurement of in mixed

cultures, Monod (1949) developed a model equation that assumes
that the rate of substrate utilization, and therefore the rate of
biomass production, is limited by the rate of enzyme reactions
involving the substrate compound that is in shortest supply relative
to its need. The Monod equation is
 The growth rate of biomass follows a hyperbolic function .
Two limiting cases are of interest in the application of Equation to wastewater
treatmentsystems.
In those cases where there is an excess of the limiting substrate, then S>>K s
and the growth rate constant is approximately equal to m . Equation 22-14
then becomes zero order in substrate.
At the other extreme, when S <<< K s , the system is substrate-limited and the
growth rate becomes first-order with respect to substrate.
Monod growth rate constant as a function
of limiting food concentration
 The above equation assumes only growth of
microorganisms and does not take into account natural
die-off.
It is generally assumed that the death or decay of the
microbial mass is a first-order expression in biomass and
hence equation becomes

where kd endogenous decay rate constant, d-1.

 If all of the substrate in the system were converted to biomass, the rate of substrate
utilization ( dS / dt ) would equal the rate of biomass production. Because of the
inefficiency of the conversion process, the rate of substrate utilization will be greater than
the rate of biomass utilization, so

where r su substrate utilization rate or rate of substrate concentration change due to

utilization.
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