CHAPTER III

BIOTECHNOLOGY APPLICATION IN WASTEWATER

TREATMENT

General aims
1. Students understand the application of biotechnology in wastewater treatment
2. Students understand the development and application of wastewater treatment

Specific Aims
1. Students understand role of microorganisms in application of biotechnology in
wastewater treatment
2. Students understand classification of microorganisms involved in wastewater
treatment
3. Students understand bacterial biochemistry, decomposition of waste, and population
of dynamics involved in wastewater treatment
4. Students understand methane production in anaerobic decomposition
5. Students understand several types of equipment in wastewater treatment

3.1. Introduction

In urbanized countries, vast amounts of industrial and domestic wastewater are

treated biologically. At present, wastewater is treated primarily by the classic, aerobic,
activated sludge process with surface aeration (Figure 3.1) or with oxygen supplied by
forced air. In another type of aerobic treatment process (the trickling filter), a bed of
stones or sand is used and the waste water is allowed to trickle down over the
supporting medium, the organisms growing attached to the surfaces of the filter bed and
oxidizing the organic compounds in the waste material. In another process, anaerobic
digestion is used (Figure 3.2), occasionally directly on raw wastewater or more fre-
quently on the solid material (sludge) obtained from sedimentation in the aerobic
treatment processes.
Aerobic systems have the following disadvantages: open construction which
restricts process control, uncontrolled populations of organisms, environmental
pollution due to odor emission, and fog formation.
Figure 3.1 Activated sludge process: above, aeration basin; below, oxidation ditch

A further critical problem is the phenomenon of bulking in sludge, which

inhibits the settling process in the clarification basins. Bulking is due to the
development of filamentous microorganisms that do not settle well. Although bulking
can be controlled in some cases by introduction of flotation processes, in domestic
wastewater this technical solution is too expensive. Therefore, a microbiological
solution to the bulking problem must be found so that a satisfactory clear effluent can be
delivered from the final settling basin.

Figure 3.2 Sludge Digester

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 One of the most encouraging developments over the past several decades has
been the remarkable decrease in organic material in industrial effluents. This has
resulted from extensive application of research and development efforts and
considerable expenditures for new treatment installations.

3.2 Wastewater Microbiology

3.2.1. Role of Microorganisms

The stabilization of organic matter is accomplished biologically using a variety

of microorganisms. The microorganisms are used to convert the colloidal and dissolved
carbonaceous organic matter into various gases and into cell tissue. Because cell tissue
has a specific gravity slightly greater than that of water, the resulting tissue can be
removed from the treated liquid by gravity settling.

It is important to note that unless the cell tissue produced from the organic
matter is removed from the solution, complete treatment will not be accomplished
because the cell tissue, which itself is organic, will be measured as BOD in the effluent.
If the cell tissue is not removed, the only treatment that will be achieved is that
associated with the bacterial conversion of a portion of the organic matter originally
present to various gaseous end products.

3.2.2. Classification of Microorganisms

By kingdoms. Microorganisms are organized into three broad groups based on

their structural and functional differences. The groups are called kingdoms. The three
kingdoms are animals, plants, and protista. Representative examples and characteristics
of differentiation are shown in Figure 3.3.

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. Energy must be obtained from outside of the cell to
enable synthesis to proceed. Our goal in wastewater treatment is to convert both the

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carbon and the energy in the wastewater into microorganisms that we can remove from
the water by settling. Therefore, we wish to encourage the growth of organisms that use
organic material for both their carbon and energy source.

Figure 3.3 Classification of Microorganisms by Kingdom

If the microorganism uses organic material as a supply of carbon, it is called

heterotrophic. Autotrophs require only CO2 to supply their carbon needs.

Organisms that rely only on the sun for energy are called phototrophs. Chemotrophs
extract energy from organic or inorganic oxidation/reduction reactions. Organotrophs
use organic materials, while lithotrophs oxidize inorganic compounds.

By their relationship to oxygen. Bacteria also are classified by their ability or

inability to utilize oxygen as a terminal electron acceptor in oxidation/reduction
reactions. Obligate aerobes are microorganisms that must have oxygen as the terminal
electron acceptor. When wastewater contains oxygen and can support obligate aerobes,
it is called aerobic.

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 Obligate anaerobes are microorganisms that cannot survive in the presence of
oxygen. They cannot use oxygen as a terminal electron acceptor. Wastewater that is
devoid of oxygen is called anaerobic. Facultative anaerobes can use oxygen as the
terminal electron acceptor and, under certain conditions, they can grow in the absence
of oxygen.

Under anoxic conditions, a group of facultative anaerobes called denitrifiers

utilizes nitrites (NO2) and nitrates (NO3) as the terminal electron acceptor. Nitrate
nitrogen is converted to nitrogen gas in the absence of oxygen. This process is called
anoxic denitrification.

By their preferred temperature regime. 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°C are called psychrophiles.
Mesophiles grow best at temperatures between 25 and 40°C. Between 45 and 60°C, the
thermophiles grow best. Above 60°C, stenothermophiles grow best. The growth range
of facultative thermophiles extends from the thermophilic range into the mesophilic
range. These ranges are qualitative and somewhat subjective. You will note the gaps
between 20 and 25°C and between 40 and 45°C. Do not make the mistake of saying that
an organism that grows well at 20.5°C is a mesophile. The rules just aren't that hard and
fast. Bacteria will grow over the range of temperatures and will survive at a very large
range of temperatures. For example, Escherichia coli, classified as mesophiles, will
grow at temperatures beween 20 and 50°C and will reproduce, although very slowly, at
temperatures down to 0°C. If frozen rapidly, they and many other microorganisms can
be stored without a significant death rate.

3.3. Some Microbes of Interest in Wastewater Treatment

Bacteria. The highest population of microorganisms in a wastewater treatment

will belong to the bacteria. They are single-celled protists. Conditions in the treatment
plant are adjusted so that chemoheterotrophs dominate. No particular species is selected
as "the best."

Fungi. For our purpose, we define fungi as multicellular, non-photosynthetic,

herotrophic protists. Fungi are obligate aerobes reproduced by a variety of methods

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including fission, budding, and spore formation. They form normal cell material with
one-half the nitrogen required by bacteria. In a nitrogen-deficient wastewater, they are
found to predominate over the bacteria.

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.

Protozoa. For our purpose, we may say that protozoa are single-celled animals
reproduced by binary fission (dividing in two). Most are aerobic chemotrophs, and they
often consume bacteria. They are desirable in wastewater effluents because they act as
polishers in consuming the bacteria.

Rotifers and crustaceans. Both rotifers and crustaceans are aerobic,

multicellular chemoheterotrophs. The rotifer derives its name from the apparent rotating
motion 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 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 underloaded
lagoons. Their presence is indicative of a high level of dissolved oxygen and a very low
level of organic matter.

3.4. Bacterial Biochemistry

3.4.1. Metabolism. The general term that describes all of the chemical activities per-
formed by a cell is metabolism. This in turn is divided into two parts: catabolism and
anabolism. Catabolism includes all the biochemical processes by which a substrate is
degraded to end products with the release of energy. In wastewater treatment, the
substrate is oxidized. The oxidation process releases energy that is transferred to an
energy carrier which stores it for future use by the bacterium (Figure 3.4).

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 Figure 3.4 General Scheme of Bacterial Metabolism

Anabolism includes all the biochemical processes by which the bacterium

synthesizes new cells. The synthesis process is driven by the energy that was stored in
the energy carrier.

3.4.2. Decomposition of Waste

The type of electron acceptor available for catabolism determines the type of
decomposition (that is, aerobic, anoxic, or anaerobic) used by a mixed culture of
microorganisms. Each type of decomposition has peculiar characteristics which affect
its use in waste treatment.

Aerobic decomposition. From our discussion of bacterial metabolism you will

recall that molecular oxygen (O2) must be present as the terminal electron acceptor for
decomposition to proceed by aerobic oxidation. As in natural water bodies, the oxygen
is measured as DO. When oxygen is present, it is the only terminal electron acceptor
used. Hence, the chemical end products of decomposition are primarily carbon dioxide,
water, and new cell material (Table 3.1). Odiferous gaseous end products are kept to a
minimum. In healthy natural water systems, aerobic decomposition is the principal
means of self-purification.

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Table 3.1 Waste Decomposition End Products

A wider spectrum of organic material can be oxidized aerobically than by any

other type of decomposition. This fact, coupled with the fact that the final end products
are oxidized to a very low energy level, results in a more stable end product (that is, one

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that can be disposed of without damage to the environment and without creating a
nuisance condition) than can be achieved by the other oxidation systems.

Because of the large amount of energy released in aerobic oxidation, most

aerobic organisms are capable of high growth rates. Consequently, there is a relatively
large production of new cells in comparison with the other oxidation systems. This
means that more biological sludge is generated in aerobic oxidation than in the other
oxidation systems.

Aerobic decomposition is the method of choice for large quantities of dilute

wastewater (BOD5 less than 500 mg/L) because decomposition is rapid, efficient, and
has a low odor potential. For high strength wastewater (BOD5 is greater than 1,000
mg/L), it is not suitable because of the difficulty in supplying enough oxygen and
because of the large amount of biological sludge that is produced. In small communities
and in special industrial applications where aerated lagoons are used, wastewaters with
BOD5 up to 3,000 mg/L may be treated satisfactorily by aerobic decomposition.

Anoxic Decomposition. Some microorganisms will use nitrate (NO3) as the ter-
minal electron acceptor in the absence of molecular oxygen. Oxidation by this route is
called denitrification.

The end products from denitrification are nitrogen gas, carbon dioxide, water, and new
cell material. The amount of energy made available to the cell during denitrification is
about the same as that made available during aerobic decomposition. As a consequence,
the production of cells, although not as high as in aerobic decomposition, is relatively
high.

Denitrification is of importance in wastewater treatment where nitrogen must be

removed to protect the receiving body. In this case, a special treatment step is added to
the conventional process for removal of carbonaceous material. One other important
aspect of denitrification is in relation to final clarification of the treated wastewater. If
the environment of the final clarifier becomes anoxic, the formation of nitrogen gas will
cause large globs of sludge to float to the surface and escape from the treatment plant
into the receiving water. Thus, it is necessary to ensure that anoxic conditions do not
develop in the final clarifer.

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 Anaerobic decomposition. In order to achieve anaerobic decomposition molec-
ular oxygen and nitrate must not be present as terminal electron acceptors. Sulfate
(SO42-), carbon dioxide, and organic compounds that can be reduced serve as terminal
electron acceptors. The reduction of sulfate results in the production of hydrogen sulfide
(H2S) and a group of equally odiferous organic sulfur compounds called mercaptans.

The anaerobic decomposition (fermentation) of organic matter generally is

considered to be a two-step process. In the first step, complex organic compounds
fermented to low molecular weight fatty acids (volatile acids). In the second step, the
organic acids are converted to methane. Carbon dioxide serves as the electron acceptor.

Anaerobic decomposition yields carbon dioxide, methane, and water as the

major end products. Additional end products include ammonia, hydrogen sulfide, and
mercaptans. As a consequence of these last three compounds, anaerobic decomposition
is characterized by an unbelievably horrid stench!

Because only small amounts of energy are released during anaerobic oxidation,
the amount of cell production is low. Thus, sludge production is low. We make use of
this fact in wastewater treatment by using anaerobic decomposition to stabilize sludges
produced during aerobic and anoxic decomposition.

Direct anaerobic decomposition of wastewater generally is not feasible for dilute

waste. The optimum growth temperature for the anaerobic bacteria is at upper end of the
mesophilic range. Thus, to get reasonable biodegradation, must elevate the temperature
of the culture. For dilute wastewater, this is not practical. For concentrated wastes
(BOD5 greater than 1,000 mg/L), anaerobic digestion is quite appropriate.

3.5 Population Dynamics

3.5.1. Bacterial Growth Requirements

In the discussion of the behavior of bacterial cultures which follows, there is the
inherent assumption that all the requirements growth are initially present. Since these
requirements are fairly extensive and stringent, it is worth taking a moment to

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recapitulate them. The following list summarizes the major requirements that must be
satisfied:

- A terminal electron acceptor

- Macronutrients
a. Carbon to build cells
b. Nitrogen to build cells
c. Phosphorus for ATP (energy carrier) and DNA
- Micronutrients
a. Trace metals
b. Vitamins are required by some bacteria
- Appropriate environment
a. Moisture
b. Temperature
c. pH

3.5.2. Growth in Pure Cultures

As an illustration, let us examine a hypothetical situation in which 1,400 bacteria of a

single species are introduced into a synthetic liquid medium. Initially nothing appears to
happen. The bacteria must adjust to their new environment and begin to synthesize new
protoplasm. On a plot of bacterial growth versus time (Figure 3.5), this phase of growth
is called the lag phase.

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 Figure 3.5 Bacterial growth in a pure culture: The "Log-Growth Curve."

At the end of the lag phase the bacteria begin to divide. Since 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. Since 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. The population of bacteria (P) after the nth
generation is given by the following expression:

P = P0(2)n (3-1)

where P0 is the initial population at the end of the accelerated growth phase. If we take
the log of both sides of Equation 3-1, we obtain the following:

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 log P = log P0 + n log 2 (3-2)

This means that if we plot bacterial population on a logarithmic scale, this phase
of growth would plot as a straight line of slope n and intercept P0 at t0 equal to the end
of the accelerated growth phase. Thus, 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
by products build up. Because of this, at some point in time, 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 which lead to the stationary phase.

3.5.3. Growth in Mixed Cultures

In wastewater treatment, as in nature, pure cultures of microorganisms do not

exist. Rather, a mixture of species competes and survives 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.

The prime factor governing the dynamics of the various microbial populations is
the competition for food. The second most important factor is the predator-prey
relationship.

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.

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 Because of their relatively smaller size and, thus, larger surface area per unit
mass, which allows a more rapid uptake of substrate, bacteria will predominate over
fungi. For the same reason, the fungi predominate over the protozoa.

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 which in turn
die for lack of food and are then decomposed by a different set of bacteria (Figure 3.6).

Figure 3.6 Population dynamics in a closed system

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 (Figure 3.7). This condition; 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.

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 Figure 3.7 Population dynamics in an open system

3.5.4. The Monod Equation

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. In the log-growth phase, the rate expression for biomass increase is:

dX
 X (3-3)
dt

where:
dX
= growth rate of the biomass, mg/L.t
dt

µ = growth rate constant, t-1

X = concentration of biomass, mg/L

Because of the difficulty of direct measurement of k in mixed cultures, Monod

developed a model equation that assumes that the rate of food utilization, and therefore
the rate of biomass production, is limited by the rate of enzyme reactions involving the
food compound that is in shortest supply relative to its need. The Monod equation is

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 m S
  (3-4)
Ks  S

where:
 m = growth rate of the biomass, mg/L.t

S = concentration of limiting food in solution, mg/L

K s = half saturation constant, mg/L

concentration of limiting food when  = 0.5  m

The growth rate of biomass follows a hyperbolic function as shown in Figure 3.8.

Figure 3.8 Monod Growth Rate Constant as a Function of Limiting Food

Concentration

Two limiting cases are of interest in the application of Equation 3-4 to

wastewater treatment systems. In those cases where there is an excess of the limiting
food, then S >> KS and the growth rate constant, µ, is approximately equal to µm.
Equation 3-3 then becomes first-order in biomass. At the other extreme, when S << KS,
the system is food-limited and the growth rate becomes zero-order with respect to
biomass, that is, it is independent of the biomass.

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 Equation 3-4 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 Equations 3-3 and 3-4 are
expanded to:

dX  m SX
  kd X (3-5)
dt KS  S

where kd = endogeneous decay rate constant, t-1.

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

dS 1 dX
  (3-6)
dt Y dt

where Y = decimal fraction of food mass converted to biomass

= yield coefficient, mg/L biomass per mg/L food utilized

Combining equation 3-3, 3-4, and 3-6,

dS 1  m SX
  (3-7)
dt Y K S  S

Equations 3-5 and 3-7 are a fundamental part of the development of the design
equations for wastewater treatment processes.

In this chapter, new treatment processes will be discussed which have the potential for
increasing the efficiency of wastewater treatment.

3.5 Treatment Processes

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 Conventional wastewater treatment involves the use of microorganisms which
develop naturally within the wastewater treatment system, no attempt being made to
optimize the organisms involved. An approach which may have some potential for
increasing the efficiency of the wastewater treatment process is to inoculate the system
with microorganisms which have been specially selected for the particular wastewater
treatment process. In analogy with their use in food fermentations, such organisms
might be called "starter cultures". Although starter cultures might find use in the
treatment of domestic wastewater, it seems more likely that they will find use in the
treatment of special or unusual industrial wastes or in the treatment of accidental spills
of industrial chemicals. Such wastes often cannot be channeled into ordinary treatment
plants because their toxicity and lack of biodegradability cause significant damage to
the non-adapted organisms. Another way in which such starter cultures might be used is
in shortening the start-up time that is generally required after a wastewater treatment
plant is shut down for one or another reason. Before the system can become fully
operative once again, the optimal bacterial culture mixture must be re-established; this
usually requires a week-long enrichment process. A starter culture could be expected to
shorten this start-up time. In the United States, starter cultures resembling those of the
dairy industry have been developed and are suitable for a variety of special applications
in wastewater treatment, such as tank cleaning, pipeline cleaning, start-up of city
purification plants, and breakdown of special substances contained in wastewater.
Bacteria from cold habitats have been isolated which degrade alkanes and
aromatic compounds at 0-15°C in saline habitats; these could be useful in the
degradation of oil spills in the ocean. Mixed cultures which metabolize DDT, poly-
chlorinated diphenols, and phenols or which possess high protease, lipase, or cellulase
activity are also on the market. If special operating conditions are desired, such as
growth at pH < 5.0 or > 9.0, these requirements can also be met with selected
enrichment cultures.
A patented process has been developed with a strain of Pseudomonas putida
containing plasmids which code for the breakdown of octane, xylene, metaxylene, and
camphor. Starter cultures have also been used to deodorize animal excrements.

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 In 1978, the starter-culture industry grossed $2-4 million in the United States,
but the potential total value is estimated at $200 million. Twenty manufacturers produce
single strains or mixed cultures under sterile conditions in vessels up to 10 m 3 capacity.
The culture conditions used are determined by how the cultures are to be used, in order
to ensure that the cells are fully induced. Research in this field is still in its infancy and
scientific studies on the breakdown of individual compounds are underway, but
practical applications are not yet widespread.

3.5.1. Aerobic wastewater Treatment Airlift Process

A disadvantage of customary aerobic wastewater treatment by the activated

sludge system is the low efficiency of oxygen transfer and the large amount of space
required by the installation. Because these installations are open to the atmosphere,
another problem is the odor which they emit. Tower reactors have been developed in
which aeration and oxygen utilization are improved and the efficiency of the overall
process increased. Both tube reactors and airlift fermentors have been used. These
installations are favor-able economically because of 30% less space required, 20%
lower investment costs, and 20% less energy costs.

Figure 3.9 Bayer Biotower

The British chemical company ICI uses a tubular loop reactor which is
embedded 100 meters into the ground. Two German companies, Bayer (Biotower) and
Uhde/Hoechst (Bio-high Reactor) have constructed bioreactors 30 m in height. In these

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systems, the circular settling basins are located around the top rim of the bioreactor
(Figure 3.9). The dimensions of this type of activated sludge fermentor allow a
considerably better oxygen-transfer efficiency. Table 3.2 shows performance data from
various systems of the Uhde/Hoechst type and Table 3.3 shows data for the Bayer
system.

Table 3.2 Data from an Activated Sludge System (Uhde/Hoechst)

Water depth (m) 4 8 20
02 Saturation 13 17 28
O2 saturation (g O2/m3)
Oxygen yield (%) 8-15 15 -30 40-60
Air required (Nm3/kg O2) 42-23 23 -11 9- 6
Energy-specific O2 input (kg O2/kWh) 2- 2.5 2.5- 3 3- 3.5
Bioreactor diameter (m) 18 13 8
Reactor surface area volume for 1000 m3 (m2) 250 125 50

The Hoechst system has baffles to facilitate mixing. In the Bayer Biotower
process, two-component nozzles have been developed as injectors by means of which
the kinetic energy of the liquid stream is utilized to disperse air in small gas bubbles in a
manner similar to a water spray pump (Figure 3.10). These injectors are installed in
groups of four (injector clusters) above the bottom of the tower reactor (1 injector/1-2
m2 bottom surface).

3.5.2. Aeration with Pure Oxygen

Aeration of activated sludge treatment facilities with pure oxygen has been tried
in both the United States and the Federal Republic of Germany. The objective has been
to increase the efficiency of oxygen transfer, since the loading capacity of a
conventional purification plant and the size of the microbial population are limited by
the low solubility of dissolved oxygen.

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Table 3.3 Parameters for a Bayer Biotower in continuous operation

Process Parameters
Screening, neutralization, primary
Pretreatment sedimentation

Bioreactors 4 parallel, rubber-lined tanks (26x30m)

with 13,600 total volume
Aeration 22,000 m3/h
Wastewater From industry 90,000 m3/d with 95 tons BOD/d

From domestic sources 70,000 m3/d with 14 tons BOD/d

Residence time
Treatment capacity 105 tons BOD/d
Residence time Treatment capacity Oxygen
requirements 120 tons O2/d

Pure oxygen has 4.8 times the partial pressure of oxygen from the air, so that
aeration with pure oxygen markedly increases the oxygen content of the activated
sludge system. Dissolved oxygen in such a system is 90-95% utilized, and the off gas
(1% of that from conventional aeration) can subsequently be treated chemically or
thermally in the closed system in order to prevent unpleasant odors.
High loading capacity in the oxygen process allows smaller aeration tanks and
smaller settling basins. Due to the smaller amount of sludge produced, the costs of
facilities for further treatment are also reduced. On the negative side is the higher capital
investment, the more complicated installation, the necessity for careful control of the
process parameters, and the need for highly trained personnel.
In both domestic and industrial installations systems are in use that employ pure
oxygen. These systems are especially used in the paper, chemical, and food industries.

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 Figure 3.10 Bayer injector (Zlokarnik, 1985)

The Unox system is one such example. The first aeration stage consists of 2
channels, each of which is connected in a cascade arrangement to three bioreactors
(Figure 3.11) with capacities of 1500, 750 and 730 m 3. In the first bioreactor (activated
sludge basin), pure oxygen from an air separation plant is forced in via injectors. In this
stage, 90-93% of the organic, decomposable material is eliminated.

Figure 3.11 Diagram of an Activated Sludge System using Pure Oxygen

The CO, produced is collected along with the residual oxygen and treated at
1000°C. In the second treatment stage (settling basin), the accumulating sludge is

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pumped back into the activated sludge basin. The waste water flows through this second
treatment stage to remove material which is not easily broken down (Figure 3.12). After
a third treatment stage, the clarified waste water proceeds to the receiving stream. Table
3.4 gives performance characteristics using the Unox process.

3.5.3. Methane Production

Methane, or natural gas, is a microbial product of the anaerobic decomposition
of organic matter. Methanogenesis is a widely used process in organic waste disposal,
primarily because the methane gas, being insoluble, is readily removed from the
treatment system. Since it is a fuel, the methane produced in the treatment system can
also be used as a source of energy. In some agricultural situations, the methane formed
is a significant energy source (biogas) and the main goal of the treatment process is the
maximization of methane yields. Anaerobic installations for wastewater treatment are
called digestors, because the primary process taking place is the digestion of the organic
material to soluble and gaseous constituents. In most waste treatment systems, however,
even though the methane may be used as a fuel, the main goal has not been the
production of methane as an energy source but rather the stabilization of the sludge so
that it can be safely disposed of in the environment. In such systems, little attention has
been given to the optimization of the methane production process. In one study of 275
wastewater treatment systems in Western Europe, only a small fraction of the methane
produced was distributed to other users, although 75% of the gas produced was used as
an energy source within the treatment plants themselves.

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Figure 3.12 Methanol Breakdown in a Two-stage Purification Plant using Pure
Oxygen

In comparison to aerobic processes, anaerobic process has some significant

advantages. Energy expenditure for aeration is not required, saving considerably on
operating costs. The organic waste is converted almost quantitatively (>90%) to
methane and CO2, Biomass formation is low, so that the cost of sludge disposal is
lower. Also, odor problems are decreased since the process is carried out in closed
reactors. Finally, the methane produced can be utilized as an energy source.
However, the anaerobic process is not suitable for the more dilute liquid wastes that are
primarily treated by the aerobic activated sludge processes.
The methanogenic bacteria constitute a unique group of organisms which are the
final and key link in the breakdown of organic matter in the anaerobic food chain. The
methanogenic bacteria are able to utilize only a restricted group of substrates for the
production of methane, including: acetate, methanol, formate, and H2 + CO2. In waste
disposal systems, about 75% of the methane is derived from acetate and most of the rest
from H2 + CO2. The starting materials of the anaerobic decomposition process are com-
plex polymeric materials such as cellulose, starch, fats, and proteins, none of which the
methanogenic bacteria are able to use. The methanogenic bacteria are consequently

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dependent upon other anaerobic fermentative organisms for the initial breakdown of the
substrates and the production of acetate and H2 + CO2.

Table 3.4 Data from a Unox purification plant with pure oxygen

Amount of wastewater 20,000 m3/day

Chemical oxygen demand (COD)-breakdown capacity 61 tons/day
Biological oxygen demand (BOD)-breakdown capacity 34 tons/day
Oxygen input
Activation stage 1 51 tons/day
Activation stage 2 7 tons/day
Volume
Activation stage 1 6200 m3
Activation stage 2 3200 m3
Intermediate purification: 4400 m3
Final purification 4800 m3
Time required
Activation stage 1 7.5 h
Activation stage 2 3:8 h
Intermediate purification: 5.3 h
Final purification 5.8 h
Breakdown activity
max. 80% COD
Activation stage 1
max. 93% BOD5
max. 7% COD total 87%
Activation stage 2
max. 5% BOD5 total 98%
Sludge load
Activation stage 1 0.7 kg BOD5/kg dry weight.d
Activation stage 2 0.2 kg BOD5/ kg dry weight.d
Space requirement
Activation stage 1 5.6 kg BOD5/m3.d
Activation stage 2 0.8 kg BOD5/ m3.d
Excess sludge 25% of the BOD5 load

Three groups of microorganisms participate in the anaerobic process. The first

group breaks down the original organic material (starches, fats, proteins) into organic
acids (propionic acid, butyric acid, acetic acid, lactic acid, and valeric acid), alcohol, H 2,
and CO2. Since one of the constituents of the initial load, lignin, is not broken down
anaerobically, any lignocelluloses in the starting material are metabolized slowly. The

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group of bacteria involved in this initial breakdown of organic matter includes obligate
anaerobes such as the clostridia and facultative anaerobics such as the streptococci and
enteric bacteria.
The second group of bacteria converts the longer-chain fatty acids (for instance,
propionic and butyric acids) and alcohols to acetic acid, H2, and CO2. This conversion is
endergonic at pH 7 and can only occur if coupled with exergonic reactions. Thus, the
reaction only occurs in mixed cultures. For example, ethanol is converted to acetic acid
and H2 by one bacterium and in a second reaction a methane bacterium utilizes the H 2,
thus pulling the reaction:

CH3CH2OH + H2O  CH3COO- + H+ + 2H2

4 H2 + CO2  CH4 + 2 H20

The third group of organisms is the extremely anaerobic methanogens. As noted,

only a limited range of substrates are used by these bacteria. Most methanogens will use
H2 as energy source and CO2, as carbon source and electron acceptor. A few species of
morphologically diverse groups (cocci, sarcinae, and spirillae) also metabolize formic
acid, methanol, or methylamine. The most important organic substrate of methanogens
is acetic acid, which is fermented in a reaction that is only weakly favorable en-
ergetically:

CH3COO- + H+  CH4 + CO2 ∆GO = - 37 kJ

Because of this, these bacteria grow very slowly and are the most critical organisms in
the mixed culture of the anaerobic digestor.
Because of the low biomass formed and the low energy yield, the rate of the
digestion process is low, the average residence time in the digestor being greater than 20
days. Immobilization of microorganisms in a fixed-bed reactor can be used to increase
greatly the biomass concentration. Solutions with very high organic loads (chemical
oxygen demand >3 g O2 consumption/1) can be treated with such fixed-bed reactors
within a few hours. Some studies have been carried out using porous sintered glass as
carrier material for fixed-bed reactors. In a 1000L reactor the biochemical oxygen

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demand (BOD) was reduced from 6.4 kg/m3 to 1.3 kg/m3 in 4.8 hours. In another
installation for treating high-concentration wastes from a fermentation plant, a two-
stage process was used, the first reactor being used for acid production, the second for
methanogenesis. Both reactors were 380 m3 in volume and sand was used as the carrier.
Waste volumes of 150-200 m3 /h were treated within 1 - 1.5 hours with a removal
efficiency of 30 kg COD/m3-day.
In the United States, some studies have been done to produce methane from the
biomass of the water hyacinth, a plant which causes considerable problems by excess
growth in canals and streams in the warmer parts of the country. Under optimal
conditions, about 2 tons dry mass per day and per hectare could be treated, yielding
theoretically 800 m3 biogas.
When optimizing methanogenesis, it is important to note that the rate is affected
by the nature of the substrate as well as by the temperature (Table 3.5).

Table 3.5 Biogas production in relation to temperature

Temperature % Dry weight Time 1 Methane/g dry
Substrate
°C in input (h) weight
35-40 2.1 23 0.488*
Sludge 50 2.1 9 0.488*
60 2.1 12.5 0.488*
30 3 33-50 0.371
Sludge
50 3 20 0.368
Waste and 35 3 30 0.288*
sludge 50 3 10 0.369*
60 3 4 0.455*
55 2 3 0.16
9 0.19
Cow
55 6 3 0.16
manure 9 0.19
55 10 3 0.10
9 0.19

With mixed sludge, as much as 600 L biogas per kg dry organic matter has been
produced. The gas consists of about 60 - 70% methane, about 25 - 30% CO2, and the
rest H2 and N2. Developing countries represent the most promising areas for further
development of biogas production, since energy is often in short supply and waste

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disposal can often not be done by the elaborate processes used in developed countries.
In such cases, the efficiency of the process is less critical than the energy produced;
biogas production in developing countries can often be accomplished in simple
installations with little requirement for complicated engineering skills. Biogas can also
be used as a motor fuel in those regions where petroleum sources are expensive or in
short supply.

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