BIOLOGICAL TREATMENT

METHODS
 DEFINITIONS
• BIOLOGICAL TREATMENT is the degradation of organic waste by the action of
microorganisms.
• Degradation alters the molecular structure of organic compounds, and the degree of
alteration determines whether biotransformation or mineralization has occurred.
• Biotransformation refers to the simplification of an organic compound to a daughter
compound.
• Mineralization is the complete breakdown of organic molecules into cellular mass,
carbon dioxide, water, and inert inorganic residuals.
• Biotransformation is partial degradation and mineralization is complete degradation.
 BASIC MICROBIOLOGY
• Several microbiological factors affecting biological treatment include:

1. Energy and substrate sources

2. Enzymatic processes
3. Substrate biodegradability
4. Inhibition and toxicity
5. Microbial community
 ENERGY AND SUBSTRATE SOURCES

• Any form of living matter requires energy and carbon for growth and maintenance

Types of microorganisms based on their energy and carbon sources

S.No. Classification of Energy source Carbon source
microorganisms (substrate)
1 Autotrophic
1a Photoautotrophic Light Carbon dioxide
1b Chemolithotrophic Oxidation-reduction reactions of Carbon dioxide
inorganic compounds
2 Heterotrophic Oxidation-reduction reactions of Organic carbon
organic compounds

• Biological treatment is the direct result of heterotrophic metabolism in which organic

waste is the substrate, i.e., the waste serves as a source of carbon and energy to living
organisms (most commonly heterotrophic bacteria).
 ENZYMATIC PROCESSES
• For degradation to occur, the waste (as substrate) must first come into contact with a bacterial
cell’s outermost coating. This action initiates a series of metabolic steps in the degradation of
organic waste.
• The first step consists of transport of the substrate into the cell which can occur via three
methods:
a) Extracellular enzyme complexation
b) Liquefaction
c) Direct transport
o Upon contact of bacteria with substrate, extracellular enzymes produced by the bacteria will
form complexes with substrate molecules. These complexes allow the substrate to pass
through the cell wall. Once inside the cell, intracellular enzymes will further complex with the
substrate to catalyze other reactions necessary to obtain energy and to build new cellular
material.
o Some organic chemicals are too large to be transported into the cell and liquefaction, i.e.,
enzyme-mediated transformation takes place on the surface of the cell. Also, some organic
chemicals can pass into bacterial cells directly without complexing with enzymes.
Metabolic steps in degradation of waste
• Enzymes are large protein molecules composed primarily of amino acids. They lower the
energy required to activate a reaction and thereby speed up biological activity. Since
enzymes act as catalysts, they are not used up in the process but can be reutilized.

• A cell operates two critical types of metabolic processes, known as anabolic (cell building)
and catabolic (energy releasing) processes. The oxidation-reduction reactions which
release energy involves a transfer of electrons from organic carbon as it is oxidized to a
higher state. To complete the reaction, electron acceptors are needed which can be
oxygen, nitrate, sulfate. The energy released by catabolic processes is captured and
transformed by adenosine triphosphate and adenosine diphosphate (ATP-ADP) energy
coupling cycle to support cell maintenance and cell building activities.

• The anabolic processes produce protoplasm as an end product which is composed of 75-
80% water. The remaining solid material is 90% organic and 10% inorganic. The organic
portion is represented by formula 𝐶5 𝐻7 𝑂2 𝑁. The inorganic cellular material contains P, Na,
Ca, Mg, S, K, Fe, and various trace metals.
 SUBSTRATE BIODEGRADABILITY

• Most synthetic organics are biodegradable, but some specific compounds may resist
biodegradation (such compounds are called recalcitrant or refractory) or their
degradation occurs so slowly as to make biological treatment inefficient.

• The inherent biodegradability of a chemical depends to a large extent upon its molecular
structure. Small changes in the configuration of a biodegradable molecule may result in
persistence.

• The following conditions, either individually or in

combination, are associated with recalcitrant or
persistent compounds:
a) Halogenation
b) Large number of halogens
c) Highly branched
d) Low solubility in water
• Some general conclusions regarding the ability of microorganisms to degrade specific
classes of organic chemicals:

✓ Typically, straight-chain aliphatic compounds are easily degraded. However, introduction

of extensive branching of alkane substituents results in sterically hindered configurations
that often are resistant to degradation. Also, unsaturated aliphatics are less readily
transformed than the saturated analogues.
✓ Simple aromatic compounds are usually degradable by several mechanisms of ring
cleavage. The introduction of halogens will result in decreased biodegradability because
halogens stabilize the ring. The degree of decreased biodegradability usually is a function
of number of halogens incorporated because biodegradation will require dehalogenation
at some stage.
✓ Even highly chlorinated aromatic compounds like PCBs can be degraded, although slowly.
Highly substituted PCB isomers are degraded much more slowly than less halogenated
derivatives, and catabolic enzymes preferentially attack and cleave the ring carrying the
lowest number of chlorines.
✓ Biodegradation of nitrogen and sulfur containing compounds is often linked to their
utilization as nutrient sources. Branched chain alkyl or aryl sulphonates are often slowly
degraded.
✓ Polymeric materials are among the most resistant to microbial attack. Nevertheless,
microorganisms have been isolated that will utilize nylon polymer or plystyrene as the
sole source of carbon and energy.

➢ Many exceptions exist to these guidelines. An engineer considering biological treatment

of a hazardous substance should depend on literature studies or testing of that specific
substance. For substances whose biodegradability has not been reported in the literature,
tests include BOD, ultimate oxygen demand, respirometry studies, and batch &
continuous flow lab or pilot scale treatability studies.
 INHIBITION AND TOXICITY

• Many organic and inorganic waste materials move through a progression of effects as the
concentration increases, similar to that of a dose-effect relationship.

• An organic substance that is biodegradable at one concentration can become persistent at

higher concentrations by inhibiting the growth of the microbial culture.

• At even higher concentrations, the substance can become toxic to the culture. These
effects derive presumably from the degree to which the substance overwhelms the
enzymatic systems which normally degrade it.

• Many of the inorganic nutrients necessary for the synthesis of cellular mass are toxic to
the same organisms at higher concentrations.
 MICROBIAL COMMUNITY
• Biological treatment involves complex interaction of mixed biological populations. Growth
rates and utilization of substrate is frequently higher in mixed enrichment cultures than in
pure cultures isolated from the mixture.
• In mixed microbial communities, not only are organisms that can initiate catabolism
important, but so are secondary utilizers (i.e., organisms that utilize intermediates derived
from transformation of the original substrate).

Consortium of three bacteria species for degrading chlorobenzoate

• Co-metabolism is an important example of a microbial community at work. It features the
transformation of one compound (the secondary substrate) by enzymes from
microorganisms routinely degrading another compound (the primary substrate or co-
metabolite). The microorganisms derive neither a source of carbon nor energy from the
secondary substrate; its degradation is serendipitous. The co-metabolite induces the
enzymes needed for transformation of the secondary substrate.

• Co-metabolism is a potential alternative for compounds which otherwise degrade very

slowly, such as trichloroethylene (TCE). Functioning methanotrophs (a group of
heterotrophic bacteria that use methane as their source of carbon and energy) indirectly
oxidize TCE to a TCE epoxide through MMO, which readily decomposes in water to
alcohols. Methanotrophs produce the enzyme methane monooxygenase (MMO) when
metabolizing methane (primary substrate).
 ENGINEERING FACTORS
• The engineering factors affecting biological treatment include:

1. Electron acceptor
2. Moisture
3. Temperature
4. pH
5. Total dissolved solids
6. Nutrient availability
7. Reactor design
8. Alternative carbon source
 ELECTRON ACCEPTOR

• The catabolic reactions involve transfer of electrons from the waste to an electron
acceptor, and the biological process by which this occurs is called respiration.
• In aerobic respiration, bacteria utilize oxygen as the terminal acceptor of electrons
removed from oxidized organic compounds. Typically, in the case of biotransformation,
oxygen is added to the organic molecule and hydrogen removed. Upon complete
mineralization, oxygen is reduced to water and organic carbon is oxidized to carbon
dioxide.
• The mass of oxygen required by aerobic systems can be calculated based on stoichiometry
or lab determinations.
𝐶7 𝐻12 + 10𝑂2 → 7𝐶𝑂2 + 6𝐻2 𝑂 (Catabolic metabolism)

The oxygen required to completely mineralize 1 mg alkene is calculated as:

10 (2 × 16)
= 3.33 𝑚𝑔 𝑂2 Τ𝑚𝑔 𝑎𝑙𝑘𝑒𝑛𝑒
7 × 12 + (12 × 1)
➢ For anabolic metabolism of alkene:

• Assimilation of alkene to build new cellular material:

𝐶7 𝐻12 + 5𝑂2 + 𝑁𝐻3 → 𝐶5 𝐻7 𝑂2 𝑁 + 2𝐶𝑂2 + 4𝐻2 𝑂 (1)

• Mineralization of new cellular material (endogenous respiration):

(2)
𝐶5 𝐻7 𝑂2 𝑁 + 5𝑂2 → 5𝐶𝑂2 + 2𝐻2 𝑂 + 𝑁𝐻3

Adding equations 1 and 2, we get

𝐶7 𝐻12 + 10𝑂2 → 7𝐶𝑂2 + 6𝐻2 𝑂

➢ Because the solubility of oxygen in surface water is in the range of 8 mg/L, even small
concentrations of organic waste necessitate that oxygen be supplied to aerobic treatment
operations. For providing factor of safety, a minimum DO of 2 mg/L is maintained
continuously.
• Anaerobic processes are the treatments that occur in the absence of oxygen. Here, the
organics are degraded to acetic acid, hydrogen, and carbon dioxide which are metabolized
by methanogens, thereby producing methane.

• The electron acceptor in anaerobic respiration include several inorganic oxygen bearing
compounds such as:
a) Nitrates – reduced to nitrogen (sometimes called anoxic rather than anaerobic
condition)
b) Sulfates – reduced to hydrogen sulfide
c) Carbon dioxide – reduced to methane

• The rate of anaerobic degradation is usually lower than aerobic degradation but it offers
advantages when degrading high-strength wastes. The expense of aeration is eliminated.
Less biomass is produced since the organic substrate is eventually converted to methane.

• Several wastes undergo anaerobic degradation more efficiently than aerobic process. Of
particular interest with hazardous waste is the highly specialized ability of anaerobic
process to transform halogenated compounds.
 MOISTURE

• Biodegradation requires moisture for two reasons:

a) For cellular growth because cellular tissue is 70-80% moisture

b) As a medium for movement of the microorganisms to the substrate, or vice versa, for
non-motile species

• Biodegradation in soil systems can occur at moisture levels well below saturation. It is
generally accepted that the minimum moisture content necessary for treatment of wastes
such as contaminated soil is 40% of saturation.
 TEMPERATURE

• Temperature has a major influence on growth rate. Cellular activity, particularly enzyme
systems, responds to heat so that the cell growth increase sharply with increasing
temperature until the optimum is reached.

• Increase in temperature by just few degrees can slow the cell growth by inactivating
enzymes and continued exposure to high temperatures can result in cell death.

• Unlike high temperature, low temperature is usually not lethal. Instead, the cells
eventually become dormant. A sudden decrease to a low temperature produces a much
greater reduction in cell activity than gradual decreases to the same temperature. Gradual
decreases enable the microorganisms to acclimate.
• The effect of temperature on cellular activity, and thus the rate of biological degradation,
can be expressed as:
𝑟𝑇
= 𝜃 (𝑇−20)
𝑟20
where,
𝑟𝑇 = activity rate at TC
𝑟20 = activity rate at 20C
 = temperature activity coefficient
T = temperature in C

Classification of bacteria based on temperature

Effect of temperature upon growth rate of a

particular species of bacteria
 pH

• Enzyme activity depends on pH. The amount of a particular enzyme that is in a

catalytically active state changes as pH changes, with maximum amount occurring at the
optimum pH. Accordingly, the growth of bacteria depends on pH.

• Most bacteria grow best in a relatively narrow pH range around neutrality. Die-off occurs
below a pH of 4-5 and above 9-9.5.

• Microbial activity can alter the external pH. Examples include anaearobic fermentation
which converts organic waste to organic acids, depressing pH. Nitrification also lowers pH
as does the carbon dioxide produced by aerobic degradation. If any of these effects are
not buffered, the altered pH can inhibit/kill the microbial population.
 TOTAL DISSOLVED SOLIDS

• The concentration of dissolved solids can affect liquid-based biological treatment. If the
TDS varies widely, the activity of the microbial population can decline.

• Environmental engineers have developed practical rules of thumb based on experience:

✓ TDS should not exceed 40,000 mg/L
✓ TDS should not vary by more than a factor of 2.0 over a period of few days
 NUTRIENTS
• Cellular mass contains carbon and numerous other elements. Metabolism requires these
elements as nutrients in addition to organic carbon as substrate. Phosphorus and nitrogen
are referred to as macronutrients because the synthesis of cellular tissue requires much
more of these than other nutrients.
• Frequently, nitrogen and phosphorus are not available in sufficient amounts in hazardous
waste and must be added, usually as ammonia and orthophosphate.
• The empirical formula for the organic portion of bacterial protoplasm is 𝐶60 𝐻87 𝑂23 𝑁12 𝑃,
provides a basis for calculating the theoretical amount of N and P required for synthesis of
cellular mass from waste.
• Environmental engineers commonly use, based on general experience, a rule of thumb
TOC:N:P ratio of 20:5:1.
• Rather than calculating theoretical requirements or using a general rule of thumb, the
need for nutrients should be determined as part of lab treatability tests. The specific
requirements are very much dependent upon the type of waste, availability of nutrients in
the waste, biomass yield, and design of treatment system.
• Micronutrients needed to support metabolism include S, K, Ca, Mg, Fe, and others. To
provide physical movement of ions across the cell-wall membrane into the cell cytoplasm,
the micronutrients should have a minimum concentration in water of 1-100 mg/L.
• Metabolism also requires a number of trace nutrients such as nickel, copper, zinc, various
vitamins, and others. The concentration of these nutrients can be less than 1 mg/L.
• In most cases, all micronutrients and trace nutrients can be obtained by bacteria from the
environment and do not have to be added, particularly if the waste is a contaminated soil
or one that has been in contact with soil (eg. ground water). However, an industrial
process wastewater may require their addition.
 REACTOR DESIGN AND OPERATION

• The physical design of reactor within which biodegradation occurs has a major effect on
treatment efficiency and economics.

• Important design factors include:

✓ Flow equalization
✓ Mixing regime
✓ Biomass retention time
✓ Hydraulic retention time
 FLOW EQUALIZATION

• Theoretical description of biological treatment systems assume steady-state conditions

which are not true in reality. The nature and concentration of contaminants, along with
the flow rate can vary with time. The microbial population must adjust to these variations
in a process called acclimation.

• Acclimation takes time and shock loads can upset the biomass. High concentrations of
waste are even toxic to microbial population and have inhibitory effects.

• For liquid waste with a highly variable flow rate, equalization consists of passing the
influent into a tank where the depth of water is permitted to vary. The waste is then
pumped at a constant rate from this tank.
 MIXING REGIME

• Complete mixing provides internal equalization to compensate for variations in

concentration of waste constituents.

• Another factor is that the reactor must provide sufficient mixing for the waste to come
into contact with the biomass.
 SOLIDS RETENTION TIME

• The population of waste-degrading microorganisms (biomass) in a reactor is important

because the rate of degradation shows a linear dependence upon the concentration of
biomass. This concentration can be increased by recycling, a process in which the
microorganisms generated in the bioreactor are withdrawn with the effluent and allowed
to settle in a clarifier, with the sludge returned to the bioreactor.

• Greater the SRT, greater is the concentration of biomass, and thus higher removal
efficiency.

• Another advantage of recycling is that it serves to retain the specific “degraders” that have
acclimated to the particular organic chemicals being treated.
 HYDRAULIC RETENTION TIME

• The HRT of a reactor is equal to the volume of the reactor divided by the flow rate.
Obviously, cost of treatment decreases as HRT decreases but it is constrained by the fact
that as it reduces, the ability to handle shock loads decreases.

• Also, as the HRT is decreased, the biomass must increase to achieve effective
biodegradation.

• Extremely high biomass levels are unacceptable in liquid treatment systems because of
potential problems with overloading of the clarifier, and the inability to transfer adequate
oxygen in a limited volume.
 ALTERNATE CARBON SOURCE

• A waste which contains multiple organic compounds capable of serving as sources of

carbon for biomass.

• Because of preferential differences, the biomass may degrade the compounds

sequentially, thus delaying the degradation of target compound. However, research shows
that in systems with longer SRT, the compounds are removed simultaneously, each at a
different rate.
 GROWTH KINETICS
• The performance of biological waste treatment can be measured by the rate at which
microorganisms metabolize the waste which, in turn, is directly related to their rate of
growth. Bacteria reproduce by binary fission and the time required to divide is called
generation time or doubling time.

Binary Fission
Generation Time : 20 min to
less than a day Exponential Growth

Condition: unlimited supply of food, unlimited supply of nutrients and abundance of dissolved
oxygen in water
 MICROBIAL GROWTH PATTERN

Stationary Log Death Condition: Finite amount of

Lag Exponential Phase food and nutrient supply
Phase Phase
Growth Phase
Log number of cells

Growth rate and

death rate of bacteria
are the same as the
food becomes limited

Food is limited; bacteria

metabolize own protoplasm,
death rate far exceeds the
Time production of new cells
Bacteria acclimate to the
new environment
Excess food surrounding the bacteria; rate of
metabolism and growth is a function of the ability
of microorganism to process the substrate
 Exponential Growth Phase

• Cells have abundant food and grow without limit during this phase

X = X 0e  t
where,

X is cell concentration (mass dry wt/vol)

X0 is cell concentration at start of exponential phase
μ is the specific growth rate (time-1)
t is time
• In other words, in both batch and continuous culture system, the rate of the growth of
bacteria can be represented by,

rg =
dX
= X
dt
Is it a constant?
• Specific growth rate is a function of environmental conditions for the organism, including
substrate concentration. There is a maximum rate at which organisms can grow even with
plenty of nutrients available (μmax). As substrate becomes limited, growth slows down. A
simple equation describing this behavior is called the Monod model.

m
dX
rg = = X
dt
Monod model for growth
m/2
dX  m XS = m
 s
= Ks + s
dt Ks + S 
K s is half - velocity constant (mg/L)
s is substrate concentration (mg/L)

Ks S
 CELL GROWTH AND SUBSTRATE UTILIZATION

New Cells
Food
Inorganic and organic
end products

• For a given substrate (food) the quantity of new cells produced can be defined with a
mathematical relationship
rg = −Yrsu
rg= rate of bacterial growth, mg/(L. sec)
Y= maximum yield coefficient, mass of cells formed per unit mass of BOD consumed,
mg/mg
rsu = Substrate utilization rate, mg/(L. sec)
  m XS
rg = −Yrsu and rg =
Ks + S
 m XS
or, rsu = −
Y (Ks + S )
kXS m
or, rsu = − where, k=
(Ks + S ) Y
k is defined to be the maximum rate of substrate utilization per unit mass of
microorganism

➢ In a mixed system, not all the cells are in log growth phase. Death rate are not
considered in the above expression.
 GROWTH IN MIXED CULTURES

• Growth curves for different species of microorganisms are different from each other.

• Most biological treatment processes are comprised of complex, interrelated, mixed

biological populations.

• For a mixed population, the position and shape of a particular growth pattern shall depend
on the relative abundance of the different species, food and nutrients available and also on
environmental factors such as temperature, pH, availability of oxygen, etc.

➢ Assumption: The decrease in cell mass caused by death is proportional to the concentration
of the microorganism present. The decrease in the number of microorganism is considered
to be endogenous decay.
rd = −k d X
kd = endogenous decay coefficient, time-1
X = concentration of cells (microorganisms), mg/L
 r = rg − rd
'
g
where,
rg’ = net rate of bacterial growth

 m XS
r =
'
− kd X
(K s + S )
g

rg' m S
Net specific bacterial growth rate =  = = − kd
'

(Ks + S )
g
X
rg'
Observed Yield YObs = −
rsu
 Important kinetic parameters which can be determined only through
treatability tests

𝒌𝒅
 SOLIDS RETENTION TIME

• SRT is a key factor in designing biological reactors. SRT is also known as mean cell
residence time or 𝜃𝑐 .
• A longer SRT can result in more efficient degradation, smaller reactor size, and lower cost.
If the SRT drops below the cell regeneration time, biomass will wash out faster than it
forms new cells.
• As a thumb rule, an SRT of about 20-30 days with biomass concentration not exceeding
5000 mg/L is used for ASP. A higher value of biomass will result in clarifier failing.

𝑋𝑉
𝜃𝑐 =
𝑅𝑎𝑡𝑒 𝑜𝑓 𝑏𝑖𝑜𝑚𝑎𝑠𝑠 𝑤𝑎𝑠𝑡𝑒𝑑
where,
X = biomass concentration
V = volume of the reactor
 TOXIC INHIBITION

• Many hazardous wastes can inhibit their own degradation at increased concentrations.

In both the cases, the specific

growth rate at lower
concentrations increases in a
nearly linear relationship.
However, as the concentration
increases further, the specific
growth rate of the toxic substance
slows, eventually reaching a
threshold concentration above
which growth ceases. An even
greater concentration could result
in die-off.
• If the concentration of a waste with a toxic nature is sufficiently low such that the effect of
inhibition is negligible, the equations derived from Monod kinetics are still applicable.
• However, for higher concentrations, the effect of inhibition is described by Andrews
kinetics, also known as Haldane equation.

𝑑𝑆 𝑆𝑋
=𝑘
𝑑𝑡 𝑆2
𝐾𝑠 + 𝑆 +
𝐾𝑖
where,
𝐾𝑖 = inhibition coefficient (mass/unit volume)

• From this expression, the biokinetics for various treatment reactors can be derived.
 MINIMUM SUBSTRATE CONCENTRATION

• Many cases of soil and groundwater contamination involve dilute concentrations of

hazardous waste constituents (ppb levels). While such low waste concentration may still
pose a harmful risk to receptors, it may not support a viable biomass needed for
treatment. The rate of growth decreases with decrease in substrate concentration,
eventually reaching a point where growth ceases. Further decrease in concentration will
result in biomass loss (cell death).
• Minimum sufficient substrate would occur when formation of new biomass equals loss by
endogenous decay:
YkXSmin
= kd X
K s + S min
Solving for 𝑆𝑚𝑖𝑛
𝑘𝑑 𝐾𝑠
𝑆𝑚𝑖𝑛 =
𝑌𝑘 − 𝑘𝑑
• Typical 𝑆𝑚𝑖𝑛 concentrations for aerobic treatment systems are in the range of 0.1 to 1.0
mg/L.

• In contrast, for the specific conditions of many contaminated sites, some toxic organics are
considered to pose an unsafe carcinogenic risk at concentrations as low as 0.01 mg/L.

• When concentrations of toxic organics do not exceed their 𝑆𝑚𝑖𝑛 , it is necessary to include
co-metabolism or add a primary substrate. Otherwise, effective biodegradation may not
occur.
 MATHEMATICAL APPROXIMATION FOR HAZARDOUS WASTE CASES

• In the field of hazardous waste, a simple first order approximation has been used to
describe the degradation of organic waste. This approximation stems from the following
equation,
dS kXS
=− (1)
dt (Ks + S )
• The approximation is based on two key assumptions concerning the concentrations of
substrate and biomass.
1) For the typical hazardous waste application, where the target organic chemical is at a very
low concentration, the numerical value of S is much smaller than 𝐾𝑠 . Thus, equation 1
becomes:
dS kXS
=− (2)
dt Ks
2) If the biomass concentration changes little with time, it can be considered a constant.
Thus, equation 2 becomes: 𝑘′
𝑆 𝑘
ln =− 𝑋𝑡 (3)
𝑆𝑜 𝐾𝑠
where,
t = duration of treatment (time)
𝑆𝑜 = initial substrate concentration (mass/unit volume)
𝑘 ′ = first order degradation rate constant (𝑡𝑖𝑚𝑒 −1 )
• Solving for S, equation 3 yields:
′𝑡
𝑆 = 𝑆𝑜 𝑒 −𝑘

where, S = contaminant concentration after treatment for time interval t

• Hazardous waste degradation rate constant can also be related to the half-life (𝑡1/2 ) as:
0.633
𝑘′ =
𝑡1/2
 TREATMENT SYSTEMS
Classification of biological treatment systems

A key technical factor in selecting the appropriate system is the capability to provide proper contact between the organic
constituents of the hazardous waste and the microbial population. This capacity primarily depends on the state of the waste
and its concentration.
 Appropriate treatment
system as a function of
concentration and form of
waste
 CONVENTIONAL TREATMENT

• The typical hazardous wastes treated by this method include wastewaters from industrial
processes containing toxic organic substances.
• The method consists of passing aqueous hazardous waste through a reactor containing
either suspended or attached biomass of highly active and acclimated microorganisms.
Flow can be continuous or batch, and the reactor can be operated under aerobic or
anaerobic conditions. Oxygen is added in aerobic systems by diffused or surface aeration.
• The liquid waste receives treatment both before and after biological treatment.
• Pretreatment could consist of several steps depending upon the type of waste:
a) Equalization – to dampen hydraulic surges and variable organic loading in continuous flow
systems
b) Chemical treatment – typically to precipitate toxic metals, if present
c) Physical separation – sedimentation of metallic precipitates, removal of floating material
d) Conditioning – typically to supply nutrients and adjust pH to optimum range
• After pretreatment, the liquid waste flows into the bioreactor where the dissolved organics
are metabolized by the biomass with a resulting growth of cellular mass. This can only be
achieved by biodegradation.

• The removal of a portion of organic waste occurs by methods other than biological, i.e.,
abiotic losses. Volatilization can result in significant removal of organics, especially in
aerated systems. Some organics may not be metabolized but adsorbed with colloidal
contaminants onto the biomass.

• Abiotic losses can account for significant amount of organic waste removed with no
reduction of the toxic nature of the waste. The chemical constituents have merely
transferred to other media which may require their own particular treatment.
Conventional liquid-phase treatment
 SUSPENDED GROWTH SYSTEMS

• Such systems include a bioreactor which is mixed continuously, thereby facilitating contact
between the substrate (i.e., organics in the liquid) and the suspended biomass. The
effluent washes the suspended biomass out of the bioreactor to a separation step where
the biomass is separated from the treated effluent, typically by sedimentation.
• A major portion of the settled biomass is returned to the bioreactor to maintain a proper
SRT and proper ratio between substrate and acclimated microorganisms (food:biomass).
Recycling increases the SRT beyond simple HRT. This speeds up the degradation and
reduces the reactor size.
• Typical SRT is in the range of 20-30 days. Lower SRT will result in washout and higher SRT
will interfere with the settling of sludge. Biomass should exceed 1000 mg/L to promote
proper flocculation.
• These systems are best suited for wastes containing moderate to high concentrations of
organics, as high as 5000 mg/L TOC. Low concentrations of organics do not yield enough
biomass growth.
Completely mixed, suspended growth treatment system

Treated
water
 ATTACHED GROWTH SYSTEMS

• Attached growth or fixed-film systems rely on the ability of microorganisms to attach to

surfaces of inert media. Contaminated water is passed through a bioreactor which houses
the media. The resulting microbial growth attaches to the media and forms a thick film.
The biomass remains in the reactor. Part of the effluent and biomass may be recycled.
• Trickling filter – uses rock as the inert media
• Rotating biological contactor (RBC) – consists of closely spaced vertical discs mounted to a
rotating shaft suspended horizontally just above the surface of liquid undergoing
treatment. The circular discs provide large surface area for biofilm. By rotating the discs,
the biomass is aerated and treatment occurs.
• Fluidized bed reactor – fluidized bed is achieved by growing the biofilm not on a fixed
media but on the sand particles or other inert media. The influent is introduced from
below the bed in an upflow reactor. The upflow velocity must be sufficiently high to keep
the particles suspended.
 Trickling Filter
Fluidized Bed Reactor

Rotating Biological Contactor

 AEROBIC BATCH REACTORS
• A batch reactor combines equalization, biotreatment, and sedimentation in a single tank.
These three steps are conducted sequentially, thus this type of system is called
Sequencing Batch Reactor (SBR).
• The biomass can be maintained by discharging only the clarified effluent after
sedimentation step. The time allocated to each step can be adjusted within the constraint
of the incoming flow. This provides greater flexibility than continuous systems.
• If the flow to be treated is continuous, atleast two SBRs are required: one to fill while the
other sequences through the treatment steps.
• A demonstration project treated ground water contaminated with PAHs from wood-
preserving wastes in a two-stage batch reactor system. The first tank operated for 4 days
and the second for 8 days. The first tank received concentrations of specialized bacteria
for treating low and medium molecular weight PAHs, and the second tank was prepared
with bacteria for treating high molecular weight PAHs. The percent removal for low,
medium, and high MW PAHs was > 98%. The system also reduced pentachlorophenol
concentrations by 88%.
Sequencing batch reactor operation
 ANAEROBIC SYSTEMS
• Treatment of municipal sewage (to digest excess sludge to methane and carbon dioxide),
industries with high-strength readily degradable wastewater (such as food products,
pharmaceutical, pulp and paper) – using anaerobic systems – energy saving and reduction
in sludge production.
• There have been few applications of anaerobic treatment of hazardous waste. One
application with demonstrated effectiveness is the treatment of phenols, cresols, and
other compounds associated with coal gasification.
• Anaerobic treatment has its greatest potential for treating those wastes where reductive
dehalogenation can occur more efficiently than by oxidative degradation. Examples of
such wastes include volatile, chlorinated hydrocarbons. Due to their volatility and slow
degradability under aerobic conditions, wastes such as TCE are not well suited for aerated
processes. Both problems are targeted in a co-metabolic approach using an enclosed,
attached growth process (methanogens maintained on a filter of inert media through
which TCE-contaminated water is pumped).