CHAPTER 1

FUNDAMENTALS OF
BIOLOGICAL PROCESSES
Course Learning Outcome

At the end of this chapter the student should be able:

 Explain the basis biological processes in
wastewater treatment
 Differentiate the classification of microorganism and
microbial growth
 Calculate microbial growth kinetics

 Explain the treatment train

1.1 INTRODUCTION OF BIOLOGICAL PROCESSES IN
WASTEWATER TREATMENT

 Primary treatment mostly removes large floating materials,

grits and a large portion of the suspended solid present in
wastewater.

 The colloid and dissolved solid, mainly organic nature, still

remain in the effluent.

 Fortunately, the mixed population of microorganisms, naturally

present in the wastewater, is able to utilize the colloid and
dissolved solid biodegradable organic matter as their food for
their growth and multifunction
 So to remove colloid and dissolved solid from primary effluent,
the wastewater is further treated using biomass an agent call
as secondary treatment.

 Reduction of soluble organic solid (BOD and COD) achieve by

microbial biosorption and further degradation and stabilization
by microbes.

 Microbes converts these solids into simpler end products like

H20 and CO2 and synthesize new cell known as biomass /
biological floc

 Biomass floc has a specific gravity slightly > than water and it
settles easily by settled floc is know as biological sludge
Overall objective biological treatment of wastewater:

 1) transform (i.e, oxidize) dissolve and particulate biodegradable

constituents into acceptable end product.

 2) capture and incorporate suspended and nonsettleable colloid

solid into biological floc or biofilm

 3) Transform or remove nutrients, such as nitrogen and phosphorus

 4) Remove specific trace organic constituents and compound

Physical Characteristics of Wastewater

 Main objective of biological treatment of domestic waste

water is to remove solid, mainly its biodegradable organic
component.

 The most important characteristic of wastewater is its total

solid content (floating matter, settleable matter, colloid matter
in solution)

 Other physical characteristic include turbidity, color,

transmittance, temperature, conductivity, specific gravity, and
odor.
BOD
 Biological oxygen demand

 Define as amount of oxygen required by the microorganism to

stabilize the biodegradable organic matter under aerobic
condition
 The organic content in wastewater serve as the food for the
microorganism and energy for their metabolism by its
oxidation.

COD
 Chemical oxygen demand

 Amount of oxygen required to oxidize the organic matter

present in wastewater, using dichromate in acidic condition by
chemical reaction.
 COD generally more than BOD for the same wastewater
sample because some portion non-biodegradable organic
matter is also oxidize along with the biodegradable content
due to chemical reaction
1.2 CLASSIFICATION OF MICROORGANISMS AND THEIR
ROLES

1.2.1 Role of Microorganisms

 The stabilization of wastewater accomplished biologically using variety of

microorganisms

 Microorganisms converts (oxidize) colloid and dissolved carboneous

organic matter (BOD) into simple end product and additional biomass.

 First the waste is oxidize to end product to obtained energy for cell
maintenance and synthesis new cell

 Second part, simultaneously some of the waste is converted into new

cell tissue using part of the energy release during oxidation

 Finally when organic matter is used up, the new cells begin to consume
their own tissue to obtained energy for cell maintenance
Oxidation:
COHNS + O2  CO2 + H2O + NH3 + other end products + energy

Synthesis:
COHNS + O2 + energy  C5H7NO2

Oxidation and synthesis

COHNS + O2  CO2 + NH3 + C5H7NO2 +end product

Endogenous respiration:
C5H7NO2 + 5O2  5CO2 + NH3 + 2H2O + energy
 Microorganism also remove nitrogen and phosphorus in
wastewater treatment processes.

 Biomass has a specific gravity slightly greater than water, it

can be remove from the treated liquid by gravity settling

 Complete treatment not accomplished because the biomass

which itself is organic will be measure as BOD in the effluent.

 the removal biomass from treat liquid, the only treatment

achieved is the bacterial oxidation of a portion organic matter
originally present
1.2.2 Classification Of Microorganism
(1) By Kingdom

Microorganisms are organized into five broad groups based on their

structural and functional differences. The groups are called kingdoms.
The five kingdom are animals, plants, protista, fungi and bacteria.

( 2 ) 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.

Our goal in wastewater treatment is to convert both the carbon and

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

 Organism must have sources of energy ,carbon and inorganic

(nutrient such as nitrogen, phosphorus, sulfur, pottasium ) for
synthesis of new cellular material,

Need input energy

Carbon Source
Microorganism obtain carbon for cell growth from either
organic matter or CO2.

heterotrophs autotrophs
 Algae and Sulfur reduction
photosynthetic bacteria
bacteria
heterotrophs
autotrophs

Energy source
Energy needed for cell synthesis may be supplied by light or
chemical oxidation reaction

chemotrophs

autotrophs heterotrophs

Protozoa, fungi
Nitrifying bacteria
and most bacteria
 Energy producing by chemotrophs are oxidation and reduction
involve
Transfer from electron donor oxidation Electron can
To an electron acceptor be organic or
reduction inorganic

Outside the cell (dissolve O2

Within the cell (dissolve O2)-
exogenous- external acceptor
(internal acceptor-fermentative
respiratory metabolism
metabolism)

Organism use only O2 to produce energy call obligate

Organism use nitrate/nitrite or O2 to produce energy call

facultative
( 3 ) By Their Relationship to Oxygen.

Bacteria also are classified by their ability or inability to utilize oxygen as a

terminal electron acceptor in oxidation/reductions.

Obligate aerobes are microorganisms that must have oxygen as the

terminal electron acceptor.

Obligate anaerobes are microorganisms that cannot survive in the

presence of oxygen.

Facultative anaerobes can use oxygen as the terminal electron acceptor

and under certain conditions ; they can also 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.
1.2.3 Some Microbes of interest in wastewater treatment

( 1 ) Bacteria...

Bacteria (singular, bacterium) are the simplest forms of plant life

which use soluble food and are capable of self-reproduction. They
are single-celled organisms. No particular species is selected as the
best. The highest population of microorganisms in a wastewater
treatment plant will belong to the bacteria. Bacteria are fundamental
microorganisms in the stabilization of organic wastes and therefore of
basic importance in biological treatment.

Based on nutritive requirement, bacteria are classified as

heterotrophic or autotrophic bacteria, although several species may
function both.
( 2 ) Fungi...

Fungi (singular, fungus) is a common term used to refer to

microscopic nonphotosynthetic plants, including yeasts, molds, and
bacteria. Fungi are multicelluar. Because of their importance, bacteria
are frequently excluded from the fungi classification. Their cells
require only half as much nitrogen as bacteria so that in nitrogen –
deficient wastewater, they predominate over the bacteria. The most
important group of yeasts for industrial fermentations is the genus
Saccharomyces. Saccharomyces cerevisiae is single-celled. Under
anaerobic conditions, this yeast produces alcohol as an end product.
Saccharomyces cerevisiae is facultative and performs the following
reactions :
(3) Algae...

Algae (singular, alga) are microscopic photosynthetic plants.

Because of the chorophyll 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. The process of photosynthesis is
illustrated by the equation :

The overall effect of this reaction is to produce new plant life,

thereby increasing the number of algae. By-products oxygen
results from the biochemical conversion of water. Algae are
autotrophic, using carbon dioxide as a carbon source.

Algae grow in abundance in stabilization ponds rich in inorganic

nutrients and carbon dioxide released from bacterial
decomposition of waste organics. Green algae Chlorella are
commonly found in oxidation ponds.
( 4 ) Protozoans...

Protozoa are single-celled organisms. The protozoans of significance in

biological treatment systems are strict aerobics found in activated sludge,
trickling filters, and oxidation ponds. These microscopic animals have complex
digestive systems and use solid organic matter as an energy and carbon
source. Protozoans are a vital link in the aquatic since they ingest bacteria and
algae. They are desirable in wastewater effluents because they act as polishers
in consuming the bacteria.

( 5 ) Rotifers and Crustaceans...

Both rotifers and crustaceans are animals-aerobic and multicellular. 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.
 1.3 MICROBIAL METABOLISM
Metabolism is the biochemical process performed by living organisms to
yield energy for synthesis, motility, and respiration to remain viable.

Metabolism of heterotrophic bacteria : In heterotrophic metabolism, organic

matter is the substrate (food) used as an energy source.

However, the majority of organic matter in wastewater is in the form of

large molecules which cannot penetrate the bacterial cell membrane.

The bacteria, in order to metabolize high-molecular-weight substances,

must be capable of hydrolyzing the large molecules into diffusible reactions
for assimilation into their cells.

Therefore, the first biochemical reactions are hydrolysis of :

(1)Complex carbohydrates into soluble sugar units,

(2)Protein into amino acids,
(3)Insoluble fats into fatty acids.

Heterotrophic – corbon source from organic material

Under aerobic conditions the reduced soluble organic compounds are
oxidized to end products of carbon dioxide and water (Eq. given below).

Under anaerobic conditions, soluble organics are decomposed to

intermediate end products, such as organic acids and alcohols, along
with the production of carbon dioxide and water (Eq. given below).

Many intermediates, such as butyric acid, mercaptons (organic

compounds with SH radicals), and hydrogen sulfide have foul odors.
Nitrifying bacteria : They perform the following reactions :

Sulfur bacteria : They perform the following reaction e.g. Thiobacillus :

This bacterial production of sulfuric acid occurs in the moisture of

condensation on side walls and crowns of sewers conveying septic
wastewater. Since thiobacilli can tolerate pH levels less than 1.00,
sanitary sewers constructed on flat grades inb warm climates should be
built using corrosion – resistant materials.m
Iron bacteria : True iron bacteria are autotrophs which oxidize
inorganic ferrous iron as a source of energy. These filamentous
bacteria occur in iron-bearing waters and deposit the oxidized
iron, Fe(OH)3, in their sheath. These bacteria are truly iron-
accumulating bacteria and thrive in water pipes conveying water
containing dissolved iron and form yellow or reddish – colored
slimes. When mature bacteria die, they may decompose imparting
foul tastes and odors to water.
 The sum total of all chemical processes of the cell is called
metabolism. It may separated into catabolism and anabolism

Obtaining energy Synthesis of cellular

Catabolic pathways involve the breakdown of nutrient molecules (Food: A,
B, C) into usable forms (building blocks).

In this process, energy is either stored in energy molecules for later use, or
released as heat.

Anabolic pathways then build new molecules out of the products of

catabolism, and these pathways typically use energy.

The new molecules built via anabolic pathways (macromolecules) are useful
for building cell structures and maintaining the cell.
1.4 MICROBIAL GROWTH AND KINETICS

 Microorganism consume substrate and carry out oxidation-

reduction reaction , growth occurs by the production of
additional cells.

 Thus in wastewater treatment application biomass is

produced continuously as the substrate in wastewater is
consume and biodegraded.
List summarizes major requirements that must be satisfied

1) A terminal electron acceptor

2) Macronutrients
a) Carbon to build cells
b) N2 to build cell (N:BOD5)
c) Phosphorus for ATP (energy carrier) and DNA

3) Micronutrients
a) Trace metal (Co,Cu,Fe,Mn)
b) Vitamins required by some bacteria

4) Appropriate Environment
a) Moisture
b) Temp
c) pH
d) Light
1.4.1 Bacteria Reproduction
1.4.2 Bacteria Growth Pattern

Bacteria growth in pure culture is illustrated in figure below

1.4.2 Bacteria Growth Pattern

Bacteria growth in pure culture is illustrated in figure below

Exponential growth phase

P=P0(2)n
Log P = log P0 + n log 2
Mixed culture Growth Characteristic

In wastewater treatment, as in nature, pure culture of microorganism do

not exit.

A mixture of species compete and survive within the limit set by the
environment
1.4.3 Biomass Yield

The mass new cells produce per unit substrate utilized or remove by
microorganism call cell yield

Y=g biomass produce/ g substrate utilization (consume)

For example

Aerobic heterotrophic Y= g biomass / g organic substrate

Y=
g biomass/
Anaerobic Y = g biomass / g VFA
BOD/COD
Nitrification Y = g biomass / g NH4-N oxidize

Volatile fatty acid to produce methane

.4.4 ESTIMATING BIOMASS YIELD AND OXYGEN
REQUIREMENT

 The most common approach used to define the fate

of substrate is to prepare COD mass balance

 The COD used because the substrate

concentration in wastewater can be define of its
oxygen equivalence which can conversed in the
biomass or oxidize.
Example 7.1
An aerobic complete-mix biological treatment process without recycle,
receives wastewater with a biodegradble soluble COD (bsCOD)
concentration of 500 g/m3. The flowrate is 1000 m3/d and the reactor
effluent bsCOD and VSS concentration are 10 and 200 g/m3
respectively. Determine :
A) Observed yield in gVSS/gCOD remove
B) Amount of oxygen used in gO2/g COD removed and in g/d
Growth constant

 During stabilization of organic content of wastewater, new

cells (microorganism) are also synthesized.

 Variety microorganism remove the organic matter from

wastewater during biological treatment

 and on the other hand during treatment we add organic solid

to the wastewater in the form of new cell (surplus
microorganism or surplus sludge)

 Need to maintain proper balance of biomass in wastewater

during treatment
Rate of utilization of soluble substrate

Mass of soluble substrate is decreasing with time due to substrate

utilization (-ve) can be express as:
kXS
rsu 
Ks  S

Where
rsu=rate of substrate concentration change due to utilization g/m3.d
k= maximum specific substrate utilization rate, g substrate/g microorgaism.d
X=biomass(microorganism) concentration g/m3
S=growth limiting substrate concentration in solution, g/m3
Ks=half velocity constant g/m3
Rate of soluble substrate production from biodegradable particulate organic
matter

When substrate is being used at its maximum rate, the bacteria are also growing at
their maximum rate, the maximum specific growth rate (μm)of the bacteria related to
maximum specific substrate utilization (k) as follows :
μm =kY
k=μm /Y

where
μm= maximum specific bacteria growth rate g new cells/ g cells.d
k= maximum specific substrate utilization rate g/g.d
Y= yield=g biomass produce/ g substrate utilization (consume)

substrate utilization rate is also reported in term of μm

 m XS
rsu 
Y K s  S 
In wastewater only about 20-50% of the degradable organic material enters as
soluble compounds.

Bacteria cannot consume the particulate matter directly and employ extracellular
enzyme to hydrolyzed particulate organic to soluble organic.

A rate of particulate substrate conversion is express as below

kh ( X S / X H ) X H
rXs  
K X  X S / X H 
Rate of biomass growth

rx  Y (rsu )  bX
kXS
rg  Y  bX
Ks  S
Where
r= net biomass production rate, g VSS/m3.d
Y = synthesis yield coefficient g VSS/ g bsCOD
b = endogeneous decay coefficient g VSS/ gVSS.d

Rate of Oxygen uptake

ro  rsu  1.42rg
Effect of temperature

Effect of temperature on the reaction rate of biological process is expressed as

Total volatile suspended solid and active biomass

VSS in a reactor not only consist an active biomass but also other
component such as cell debris and nbVSS in the influent wastewater
fed to biological reactor

During cell death, cell lysis occur with release of cellular material into
liquid for consumption by other bacteria

A portion of the cell mass (cell wall) is not dissolved and remains as
nonbiodegradable particulate matter is referred to as cell debris and
contribute to total VSS

The rate production of cell debris as follows

Total volatile suspended solid

The VSS production rate can be define as sum of the biomass production, cell
debris production and nbVSS in the influent

QX oi
rX T VSS  Yrsu  bX  f d b X 
V

Active Biomass

Active fraction biomass is the ratio of biomass growth divided by total VSS
Net Biomass Yield
 Ybio=rx/rsu

Observed Yield
 Yobs=rXTVSS/rsu
Example 7.5

An aerobic complex-mix treatment process is used to

treat an industrial wastewater. The amount of bsCOD
in the influent wastewater is 300 g/m3 and the influent
nbVSS conctration is 50 g/m3. The influent flowrate is
1000 m3/d, the aerobic tank biomass concentration is
2000g/m3, the reactor bsCOD concentration is 2.4
g/m3 and the reactor volume is 335m3. If the cell
debris fraction fd is 0.1, determine the net biomass
yield, the observed solid yield and the biomass
fraction in the MLVSS. Use kinetic coefficient given in
Table 7-8
1.5 Treatment train:

 A complete treatment process that include

pretreatment, primary treatment, residuals and
side-stream treatment and post treatment
considerations.
Secondary treatment is designed to substantially degrade the biological
content of the sewage which are derived from human waste, food waste,
soaps and detergent.

The majority of municipal plants treat the settled sewage liquor using aerobic
biological processes.

Microorganism require both oxygen and food to live.

The bacteria and protozoa consume biodegradable soluble organic
contaminants (e.g. sugars, fats, organic short-chain carbon molecules, etc.)
and bind much of the less soluble fractions into floc.

Secondary treatment systems are classified as fixed-film or suspended-

growth systems.

•Fixed-film or attached growth systems include trickling filters, bio-towers,

and rotating biological contactors, where the biomass grows on media and
the sewage passes over its surface
•Suspended-growth systems include activated sludge, where the biomass is
mixed with the sewage and can be operated in a smaller space than trickling
filters that treat the same amount of water.

List of alternative secondary treatment methods

Activated sludge
Aerated lagoon
Aerobic granulation
Constructed wetland
Membrane bioreactor
Rotating biological contactor
Trickling filter