CEN-202: Wastewater

Engineering
Instructor: Sudipta Sarkar
Associate Professor
Room # 215B, Cell-8954386690
Why Study Water and Wastewater Engineering as part of Civil Engineering?

CIVIL
ENGINEERING
Nature has a capacity of self-purification of the natural contaminants /
pollutions. This is called the carrying capacity of the environment.

When the pollution load is from man-made sources such as industry or it is

higher than the carrying capacity of the environment, that is

Pollution Load > Carrying capacity of the environment

The nature’s self-cleansing capacity fails to treat the pollutants

We need to engineer the natural treatment processes in such a way that

the pollutants can be treated adequately within a smaller area in
accelerated pace within a short time

Such an engineered process is called Wastewater Treatment. The engineering

involved with collection, conveyance, treatment of wastewater and disposal
of treated wastewater is known as Wastewater Engineering
 TREATED WATER
RAW WATER TREATMENT PLANT
WATER

WASTEWATER TREATMENT PLANT

TEATED
WASTEWATER WASTEWATER
About The Course CEN 202:

2. Contact Hours: L: 3 T: 1 P: 2/2

3. Examination Duration (Hrs.): Theory 3 Practical 0

4. Relative Weightage: CWS 115 PRS 15 MTE 30 ETE 40 PRE 0

S. No. Name of Books / Authors Year of
Publication
1. Davis, M. L., Water and Wastewater Engineering; Design 2013
principles and Practice, McGraw Hill Education (India) Edition
2. Hammer, M.J. and Hammer, M.J., “Water and Wastewater 2008
Technology”, 6th Ed., Prentice Hall of India.
3. Davis, M.L. and Cornwell, D.A., “Introduction to Environmental 2008
Engineering”, 4th ed. McGraw Hill.
4. Ronald Droste., “Theory and Practice of Water and Wastewater 2005
Treatment”, John Wiley & Sonc
5. McGhee, T.J., “Water Supply and Sewerage”, McGraw Hill. 1991

6. Peavy, H.S., Rowe, D.R. and Tchobanoglous, G., “Environmental 1986

Engineering”, McGraw Hill.
7. CPHEEO, Manual on Sewerage and Sewage Treatment, Ministry 2012
of Urban Development, Govt. of India

http://moud.gov.in/Manual_on_Sewerage pdf version freely available from the

website
Learning objectives

By the end of this course you will be able to:

1. Estimate the amount of sewage / wastewater to be generated from a
township/ municipality/industry.
2. Analyze and characterize sewage/ wastewater for its pollutant
load and suggest effective treatment technologies.
3. Design sanitary and storm sewers and sewer networks.
4. Use basic science of unit processes to design the unit processes
for wastewater treatment.

5. Suggest effective method for the disposal of treatment residuals.

6. Understand the needs to design alternative sanitation strategies

to suit the needs of people in underdeveloped areas.
 What you can expect of me

• Be committed to help you understand and master the

course material
• Be available for out of class assistance
• Provide competent teaching
• Treat each question or concern seriously and answer
these to the best of my ability
• Treat each of you with courtesy and respect
 What I expect of you

General Expectations
• Ask question whenever something is not clear to you
• Help each other understand the course material
• Perform all assigned reading on time
• Arrive on-time for class
• Be courteous to each other and to me
• Provide me with feedback/suggestion – on how the course and
my delivery can be improved
Revisiting Great Expectations
 Code of Conduct

• Turn off your cell phone when you enter the classroom

• Keep the internal discussions to a minimum

– Background chattering noise is distracting for the instructor and eventually to

classmates
– If you have something interesting to share, share it with the whole class

• If you are sleepy, your bed may provide a better environment for rest
than the classroom chair
Stage Feelings Thoughts Behaviour

Loss Fear Cautious Paralysed

Doubt Resentment Sceptical Resistant

Discomfort Anxiety Confused Unproductive

Danger Zone
Discovery Anticipation Creative Energised

Understanding Confidence Pragmatic Productive

Integration Satisfaction Focused Generous

CHARACTERIZATION OF WASTEWATER
 Learning Objectives
• Comprehend basic characteristics of domestic
wastewater (flow rate, BOD, SS) and apply these
characteristics to a preliminary engineering design.
• Calculate mass loadings within the wastewater
treatment system for BOD and SS.
• Calculate and interpret mass balances for mixtures of
wastewaters.
• Calculate and interpret population equivalents for
industrial wastewater.
 Sources of Wastewater
(Major Components)
1. Domestic: food, soap and detergents, Toilets (fecal and
urine), and paper.
2. Commercial: Toilets and food from restaurants and other
markets and shops.
3. Industrial: highly variable types of waste , dependent on
the type of the industry and the processes involved.
4. Agricultural field runoff: Organic waste, may contain
fertilizers, pesticides, herbicides and insecticides.
5. Runoff from streets: sand, petroleum, tars and tire
residues.
 CHARACTERISTICS OF WASTEWATER
PHYSICAL CHEMICAL BIOLOGICAL
• Color • Organic – Microorganisms and
• Temperature Biodegradable virus
• Odor and Non- - Pathogenic
• Solids biodegradable organisms
• pH
• Alkalinity
• Phosphorus
• Nitrogen
• Heavy metals
• Dissolved gases
• Priority pollutants
PARAMETERS OF IMPORTANCE

Total Solids
Turbidity
Dissolved Oxygen
Biochemical Oxygen Demand, Chemical Oxygen demand, Total
organic Carbon, Theoretical Oxygen Demand
pH
Phosphorus
Nitrogen
Toxic Inorganic Compounds
Heavy metals
Total and fecal coliform (Most Probable Number)
 PHYSICAL CHARACTERISTICS
CHARAC. SOURCE REMARKS

COLOUR DOM. & INDST., NATURAL

DECAY OF ORGANIC MATTER

ODOUR INDST. WASTE & DECAY OF Fishy, rotten egg like, rotten cabbage
ORG. MATTER like, faecal matter

TEMPERATURE DOM. & INDST. Waste water temperature > supplied

WASTEWATER, HEATING water
WITHIN PLUMBING SYSTEM

SOLIDS DOM. & INDST. WASTE, SILT 1000 L wastewater contains nearly
500 gm solids (TS, TDS, TSS, TVS, VSS);
Imparts turbidity
 Approximate Sizes of Environmental Particles
Settle-able SOLIDS
Dissolved
SOLIDS SUSPENDED SOLIDS
COLLOID

CLAY SILT SAND

ALGAE

VIRUS
BACTERIA

0.001 0.01 0.1 1 10 100

Particle Size (mm)

 SETTLEABLE SOLIDS
1 L sample is taken in an Imhoff
cone

The sample is allowed to stand

undisturbed for 2 hrs

The volume of settleable solids

can be noted from the
graduations on the cone.

The solids settled at the bottom of the

cone can be dried at 105⁰C and weighed
to find conc. In mg/l

Non-settleable solids = (Total

suspended solids - Settleable
solids)
Characterization of Solids in Wastewater

Glass Fiber Filter of 1.2

micron pore size is used
to filter the wastewater
samples

VACUUM FILTRATION APPARATUS

 Volatile solids escape
Evaporation
105 Deg C 550 Deg C

8h >1 h Inorganic
Ash

WATER VA MB MC
SAMPLE Total Solids= MB/VA Total Volatile Solids= (MB-MC)/VA

Filter
105 Deg C 550 Deg C

8h >1 h
ME MF
VB
105 Deg C
Suspended Solids= ME/VB
8h

Volatile Suspended Solids= (ME-MF)/VB

MG
Total Dissolved Solids= MG/VB
 1 L sewage
~ 1,000,000 mg

Water Solids
~ 999500 mg ~ 500 mg

Dissolved ~ ½ (~ 250 mg) Insoluble ~ ½

Ca++, Mg++, Na+, HCO3-, SO42- ,Cl-, ~ 250 mg
soluble organics

Settleable Suspended, turbid

~ 125 mg WW, ~ 125 mg
 pH
H 2O  H + + OH − K

[ H + ][OH − ]
K= = 10 −14 Being pure phase, [H2O] = 1
[ H 2O ]

K w = [ H + ][OH − ] = 10−14 Kw is the dissociation constant of water

pH = − log[ H + ]
pOH = − log[OH − ]

For pure water, [H+] = [OH-] [ H + ][ H + ] = 10 −14

[ H + ] = 10 −14 = 10 −7

pH = − log[ H + ] = − log10−7 = 7
If pH < 7, the wastewater is termed as acidic. pH>7 it is called alkaline.
Normally domestic wastewaters tend to be slightly alkaline, from 7.0 to 7.5
 Chemical Characteristics
Domestic wastewater predominantly contains dissolved organic
matters, so does many industrial wastewaters.
Bio-degradation
Bio-degradable
Organics 1. Wastewater contains organics
Non- Biodegradable such as glucose
Biodegradable fraction: 2. Microorganism utilize the organic
-Carbohydrates compounds as their food
3. Assimilation of food means
-Protein
consumption of oxygen dissolved
-Fats, Oils, and Greases in water
-Surfactants (detergents) 4. Oxygen consumed by microbes is
-Urea (agricultural run-off) replenished by mass-transfer from
air
Priority pollutants 5. If there is a mismatch in the rates,
– Industrial solvents, pesticides, etc. oxygen gets depleted, causing the
death of aquatic species

HOW TO QUANTIFY THESE DIVERSE GROUP OF ORGANIC CHEMICALS?

 Representation by effect

Conc. of specific substances – not found

Sewage – mixture of ill-defined chemicals
Representation by effect is used
The strength of the mixture is defined by some common
factor on which all the chemicals within the mixture depend.
Ex. O2 depletion from biological/chemical decomposition of the
chemical mixture.
For many organic - bearing wastes, instead of identifying 100’s
of individual compounds, it is convenient to report the effect,
in units of the mg of O2 that can be consumed/L of water.
Referred as BOD / COD
 Predominant elemental make-up of organic matter: C, H, O, N, P, S

Highest Oxidation State: +4 +5 +6

In most of the organic compounds, elements like Carbon, Nitrogen, Phosphorus

and Sulfur are not present in their highest oxidation state.
Organic compounds can undergo an unique chemical reaction, OXIDATION.
Inorganic dissolved species in wastewater cannot undergo oxidation as most of
them are present as already oxidized to the extent possible.
Oxidation is a process of transferring of electrons from one species to the other
The species which losses electron(s) from their outer electronic shell is
considered to get oxidized and the one gaining electron(s) is called an oxidant(it
itself gets reduced)

Common Oxidants are Oxygen, Chromium (VI), elemental chlorine, etc.

Oxygen requirement or oxygen demand for the complete oxidation of organics
can serve as an aggregate measurement of the amount of organics present in a
wastewater sample. .
 Specifies how much oxygen is required to completely oxidize
Oxygen Demand
the organic matter present in wastewater sample

Stoichiometry: C6 H12O6 + 6O2  6CO2 + 6H 2O

For 1 mM 180 mg/L 192 mg/L
glucose
A wastewater containing 1mM glucose solution can be quantified in two ways:
a) A solution with glucose concentration of 180 mg/L

b) A solution with theoretical oxygen demand of 192 mg/L

Derived from the chemical formula,

Experimental determination is possible; difficult when the formula or type
does not require quantification of each of organic is unknown
one of the individual organic
component; value is somewhat less
than the theoretical oxygen demand
 Steps to calculate ThOD

Example - C6H6 : Benzene : 156 mg/L

Step 1:
Write the Eq. (Oxidation to CO2 and water) C6H6 + O2 → CO2 + H2O

Step 2:
Balance the Eq. C6H6 + 7.5 O2 → 6 CO2 + 3 H2O
Sequence for balancing the no. of atoms: (i) C → 6, (ii) H → 3; and (iii) O → 7.5
78 mg/L 240 mg/L
Step 3:

156mgbenzene 1molebenzene 7.5moleO 2 32 gO 2 480mgO 2

x x x =
L 78 gbenzene molebenzene moleO 2 L
 DETERMINATION OF OXYGEN DEMAND

Measured by quantifying O2 used by the

Bio-Chemical microbes for oxidizing the organics in
Oxygen Demand (BOD) wastewater
Chemical (COD) Strong Oxidants like Cr(VI) species are
used for quick assessment

Oxidation-reduction reactions are slow processes. Microbial oxidation

reactions are essentially slow processes as they involve enzymes acting as
catalyst in the complex sub-steps in the oxidation reactions.
 Biochemical Oxygen Demand

• Amount of oxygen required by bacteria while

stabilizing the decomposable organic matter
under aerobic conditions.
• It involves the measurement of oxygen
consumed by living organisms.
 Biochemical Reaction
New Cells/CO2

Oxygen
 Biochemical Oxygen Demand
Measurement
• Take sample of waste; dilute with oxygen saturated
water; add nutrients and microorganisms (Seed: if not
present).
• Measure dissolved oxygen (DO) levels over 5 day.
• Temperature 20° C.
• In dark (prevents algae from growing), Plastic Bottle.
• Final DO concentration must be > 2 mg/L .
• Need at least 2 mg/L change in DO over 5 days.
 Simple BOD Measurement

Measure DO of Put into 20oC incubator Measure DO

the sample for 5 days or 27o C for 3 after 3 or Five
days days
 BOD Determination
Wastewater Dilution Seed Microorganism
Water
Make total volume to
300 mL Maintain a constant
Temperature; usually 20 deg
Measure Initial C; kept away from light;
Dissolved Oxygen (DOi)
concentration (mg/L)

Microbes grow in number, Utilize the food,

consumes O2, concentration of O2 falls
After a Specific Time-period,
Usually 5 days

DOi − DO f
Measure Final BOD5 d , 20 deg (mg / L) =
Dissolved Oxygen P
(DOf) concentration P = volumetric fraction of wastewater
(mg/L) used in the BOD reactor
 BOD Calculations
• If initial DO of a sample is 8 mg/L and final DO
after 5 days is 2 mg/L. What is the BOD of the
Sample.
• If initial DO of a sample is 8 mg/L and final DO
after 5 days is 0 mg/L. What is the BOD of the
Sample

46
Example: 5 ml wastewater is added to a 300 ml BOD flask. DOi = 8 mg/L
DOf = 2 mg/L after 5 days. What is the value of BOD5

P= 5 = 0.0167
300
BOD5 = 8 – 2 = 359 mg/L
0.0167

Normally Domestic sewage contains microorganisms; For industrial wastewater,

this may not be the case; there, addition of seed microorganism may be necessary.

When seed is added, we need to consider the oxygen demand generated by the
initial seed microorganism also. To evaluate this, another parallel test is run with
the seed and the dilution water but without the addition of wastewater.
 Essential Air Waste Sample, Vs
Dilution
nutrients Organic matter and no
water
Bacteria Dilution bacteria or limited
300 - Vs number of bacteria
(seed) water
300 mL
Distilled
Water B1, B2 300 mL BOD DOi, DOf
bottles
Seeded Dilution Water Seeded Blank Seeded Sample

( DOi − DO f ) − ( B1 − B2 ) f
BOD5 d , 20 deg ( mg / L) =
P
Valid only when
B1 and B2 = Initial and final DO of the control run with seed only seeded blank has
volume of seeded dil. water in the seeded sample
the same volume as
f = the combined total
volume of seeded dil. water in seeded blank
volume of wastewater in sample volume (In this case
P=
total combined volume both are 300 mL)
(total combined volume - volume of seeded dil water in sample)
= = 1− f
total combined volume
 BOD Reaction Kinetics

DO utilization curve during BOD test

DOi
DO consumed in 5 days
(Oxygen equivalent of
organics destroyed)

DOf First Order Reaction

BOD5= Oxygen equivalent of Organic destroyed in 5 days = DO

consumed in 5 days
1st few days – High conc. of org. matter present / Rapid rate of O2 depletion
As org. matter decreases /
Later- rate of O2 consumption also decreases
Last phase - O2 consumption associated with decay of bacteria those grew earlier

Assumption:
Rate of O2 consumption Proportional to conc. of degradable org. remaining at any time
1st order reaction
 BOD reaction is a first order reaction

Rate of change in reactant concentration  Amount of reactant present at

any time
dL L= Oxygen equivalent of biodegradable
− L organics present at time t, mg/L
dt
dL dL
− = kL = − kdt k =BOD rate constant, day -1
dt L
dL
Integrating we get,  L = − k  dt ln L = − kt + C

At time t = 0, L= L0 L0= Oxygen equivalent of biodegradable organics

present at t=0, mg/L
L
ln = − kt L = L0e − kt or, Lt = L0e − kt
L0
L or Lt is often known as BOD remaining at time t
 L0
Biodegradable Organics (BOD)
L0 = BODu

BODu
remaining, mg/L

BOD Exerted, mg/L

yt = L0 − Lt = L0 (1 − e − kt )

yt ( BODt )
Lt

0 5 10 15 20
Time, days
Example: In the previous example we found out that BOD5 of the
wastewater sample was 359 mg/L. Find out the ultimate BOD of the
wastewater sample. Also find out the value of BOD10 . K =0.23 per day

BOD5 = y5 = L0 (1 − e − ( 0.23 )*5 ) = 359

L0 * 0.683 = 359

L0= BODu=525.62 mg/L

BOD10 = y10 = 525.62 * (1 − e − ( 0.23 )*10 ) = 472.92 mg/L

BOD Eq. in base 10:

yt = L0 (1 − 10− Kt )

Capital K
k = 2.303 (K)
BOD rate constant (k)
This can be determined from experimental data. Ideally, if we have
more than one data point on the BOD curve, we can find out k from the
curve.
Experimental observations vary depending on variable experimental
conditions, so k is estimated from a set of experimental data, by best-
fitting a linearized BOD Curve.

Usual value of k (base e) is 0.23 per day at 20 deg C.

Value of k varies with temperature.

kT = k20 T − 20
Generally,  = 1.047
Effect of Temperature on BODt and BODu

L0 BODu

BODt varies with temperature but BODu,

being intrinsic property of wastewater,
BODt does not change
Nature of the waste

1000’s of naturally occurring organic compounds

Not all can be degraded with equal ease
Simple sugars and starches → Rapidly degrade → Large k
Cellulose → Degrade slowly → Lower k
Hair → Almost nondegradable → k ?
Sewage → k depends on relative proportion

Typical values for k:

Sample K (20 °C) k (20 °C)

(day -1) (day -1) Lower k:
Easily degradable
Raw sewage 0.15-0.30 0.35-0.70
Organic compounds
Well-treated sewage 0.05-0.10 0.12-0.23   -More completely
Polluted river water 0.05-0.10 0.12-0.23 removed during
treatment.
 K = 0.115/day
For different types of wastes having same BOD5
L0 is the same only if values of K are same.
Industrial Waste has smaller k, they have greater L0 but
same BOD5. It is expected to have greater impact on
DO in river. Smaller fraction of BOD exerted in 1st 5
days due to lower K. Effect of K on
BOD5 when L0
Is same.

Polluted river water:

BOD5 at 20°C = 50 mg/L, K = 0.115/day
yt = L0 (1 − 10− Kt )
L0 = 68 mg/L
River water temp. = 10°C
kT = k20 T − 20
K at 10°C = 0.032/ day
BOD5 at 10°C = 21 mg/L
Lab. Determined value of BOD5 at 20°C
Effect of K on L0 for 2 wastewaters having same BOD5 (50 mg/L) overestimates O2 consump-
tion in the river at 10°C (21 mg/L).
Graphical Determination of BOD Constants, k and L0

BOD data/ plot BOD versus time/ hyperbolic first-order curve/ asymptote- L0
Difficult to fit an accurate hyperbola to scattered data
Methods that linearize data preferred

Thomas Graphical Method (Thomas, 1950)

Relies on similarity of the series expansion of the following two functions:

thomas least square fujimoto.docx
Nitrogen Oxidation

Up to this point we assumed that only C in organic matter is oxidized.

Actually many organic compounds, such as proteins, also contain N that can be
oxidized with the consumption of O2.

However, mechanisms and rates of N oxidation are distinctly different from those of
C oxidation.
Two processes must be considered separately.

O2 consumption due to oxidation of C → called carbonaceous BOD (CBOD).

due to N oxidation → called nitrogenous BOD (NBOD)

Organisms that oxidize C to obtain energy can not oxidize N.

Instead, N is released into water as ammonia (NH3)
At normal pH, NH3 is present as ammonium cation (NH4+)
NH3 from organics + ind. wastes + agricultural runoff (fertilizers) oxidized → NO3-
by nitrifying bacteria (nitrification)

The overall reaction for ammonia oxidation :

NH4+ + 2O2 ----
microorganisms
--→ NO3
- + H2O + 2H+

Theoretical NBOD = g of O2 used / g of N oxidized = (4x16)/14 = 4.57 g O2 / g N

 Conversion of Ammonium to Nitrite (Nitrosomonas)
NH4+ + 2 O2 → Bacteria (Nitrosomonous) → NO2- + 2 H+ + H2O
BOD exerted (BOD), mg/L

Conversion of Nitrite to Nitrate (Nitrobacter)

NO2- + 0.5 O2 → (Nitrobactor) → NO3- Inhibitor for nitrogenous BOD
reaction: Methylene blue, Thiourea.
Nitrogenous
Ultimate Carbonaceous BOD (NBOD)
BOD

Carbonaceous
BOD (CBOD)

0 10 20 30
Time, days
Rate of nitrification depends on number of nitrifying organisms

Untreated sewage: Few nitrifying organisms, NBOD exerted after much of CBOD exerted
due to lag in growth

Well - treated sewage:

High conc. of nitrifying organisms and
Less lag time

Same BOD Eq.

K = 0.04 – 0.10 / day
(As for CBOD of well-treated effluent)
Same Eq. for temp. correction
LIMITATIONS OF A BOD TEST

1. Non-biodegradable organic waste is unaccounted for.

2. Wastewater with high BOD content will use up all dissolved
oxygen before the 5 days is over. Proper dilution is
necessary.
3. Industrial wastewater with no initial microorganism load
shall require inoculation of ‘seed’ bacteria. The bacteria
should be acclimatized to the wastewater, otherwise may
generate a lower BOD number.
4. The test is for a long duration. A faster test is much
required.
5. Depends on the activity of the microbes only; presence of
toxic substances such as heavy metals can inhibit the
growth of the microbes.
BOD of Selected samples

Origin 5-day BOD

(mg O2/L )
River 2
Domestic wastewater 200
Pulp and paper mill 400
Commercial laundry 2000
Sugar beet factory 10000
Tannery 15000
Brewery 25000
Cherry-canning factory 55000
Chemical Oxygen Demand (COD)
COD is measured following digestion at high temperature with strong
oxidant such as chromic acid, or sulfuric acid/potassium dichromate.

The chromate ion reacts with the COD producing a color that is
measured to determine the amount of chromate ion reacted. The
oxygen equivalence of chromate ion is known as COD.

It is a fast process. BOD test is a 5-day test. COD test takes not more
than 3 hours altogether.

The test is done with a strong oxidizing agent. So, all the organics,
whether biodegradable or non-biodegradable, shall be oxidized.

COD Biodegradable + Non-biodegradable

BOD Biodegradable
Chemical Oxygen Demand

Oxygen equivalent of the organic matter that can be oxidized by a

strong oxidizing agent (potassium dichromate) in an acidic medium.

COD > BOD5: (a) Because more compounds can be oxidized chemically
than can be oxidized biologically and
(b) Because BOD5 does not equal ultimate BOD
COD: 3 h
BOD: 5 d
 Stochiometry of COD
−2 + =3  a
Cn Ha Ob + Cr2 O + 8H → nCO2 + 2 Cr +  4  +  H2 O
7
 2
Organic Strong Sulphuric Carbon dioxide Chromic acid
Matter Oxidant Acid
Potassium
Dichromate

HEATING 2 HOURS 150 OC

Orange Green
Colored Colored

2n a b
Where: = + −
3 6 3
 Take 2.5 ml sample in COD vial

Add 1.5 ml K2 Cr2O7in it

Add 3.5 ml sulphuric acid reagent

Digest above solution in digester for 2 hr at 150 oC

Take reading in spectrophotometer

COD and BOD - Comparatives
COD Biodegradable + Non-biodegradable
BOD Biodegradable

BOD
0 1
COD

For a completely biodegradable wastewater,

BODu
= 0.9 to 1
COD
For wastewater with BODu
 0.6 , it is considered non-biodegradable
COD

Theoretically, for a completely biodegradable wastewater

BODu = COD
Theoretical Oxygen Demand
This is an oxygen demand that is calculated using stoichiometry, from
the chemical formula of a compound considering that there is a
complete degradation.
It also includes the complete oxidation of ammonia that is formed in
the first stage of reaction.

Reaction: Stage 1
Cm H n O p N r + aO2 → bCO2 + cH 2O + dNH3
Stage 2
NH 3 + 2O2 → NO3− + H 2O + H +
ThOD = BOD = COD ?
Is it possible?
Rare
Chemical composition of ALL the substances known : ThOD
Capable of being completely oxidized chemically : COD / biologically : BOD
Then, ThOD = BOD = COD
 Oxygen Demand: Definition and Notation
(All terms have units of mg O2 / L )

BOD Biochemical O2 demand – Amount of O2 utilized by microorganisms in

oxidizing carbonaceous and nitrogenous organic matter.
CBOD Carbonaceous biochemical O2 demand – BOD where electron donor is
carbonaceous organic matter.
NBOD Nitrogenous biochemical O2 demand – BOD where electron donor is
nitrogenous organic matter.
ThOD Theoretical O2 demand - Amount of O2 utilized by microorganisms in
oxidizing carbonaceous and/ or nitrogenous organic matter, assuming
all of the organic matter is subject to microbial breakdown, i.e., it is
biodegradable.
BOD5 5 – day biochemical O2 demand - Amount of O2 utilized (BOD exerted)
over an incubation period of 5 days. y5.
BODu Ultimate biochemical O2 demand - Amount of O2 utilized (BOD
exerted) when all of the biodegradable organic matter has been
oxidized. L0.
COD Chemical O2 demand - Amount of chemical oxidant, expressed in O2
equivalents, required to completely oxidize a source of organic matter;
COD and ThOD should be near equal.
TOTAL ORGANIC CARBON (TOC)
TOC is measured using a TOC analyzer. The sample is catalytically
combusted and organic carbon is quantified using infrared detection of
carbon dioxide.
 Significance of Nitrogen

NITROGEN

RECEIVING BODY WASTE WATER

- High NH3-N: Toxic to fish
TREATMENT
- Low NH3 & NO3-N : Nutrient for - Sufficient N required for optimal
algae & aquatic plants growth of microorganisms
- DO exertion if longer residence - Domestic water has good C:N:P
time (NH4+ to NO3) ratio
- Ind. WW: Some may lack adequate
N
 Nitrogen
•Indicator of sanitary condition
•Initially entire ‘N’ as Protein-N & Ammonia-N
Org-N converted to Ammonia-N
Oxidation to Nitrite-N
Oxidation to Nitrate-N
•Dominant species & implication
– Org. & Ammonia-N (TKN)
=>Fresh potentially dangerous
– Nitrate-N
=> Polluted long back,
Little public health threat
Nitrogen
An indicator of sanitary quality

Chemical tests

➢Chloride – No evidence of how recently the contamination had occurred

➢Nitrogen – Most of N originally present as organic (protein) N and ammonia
As the time progresses, org. N is gradually converted to ammonia N
Later on, if aerobic condition present, oxidation of ammonia to nitrites and nitrates occurs
For ex.
(a) Waters that contained mostly org. and
ammonia N – considered to be recently polluted
- of great potential danger
(b) Waters in which most of the N was in the form
of nitrates were considered to be polluted long
back – offered little threat to public health

Bacteriological Tests (about 1893)

provides more reliable evidence concerning

hygienic safety of water
– has eliminated the need for extended N
analysis in most water supplies
Significance of Nitrogen

(a) In receiving body:

1. In high concentrations, NH3-N is toxic to fish.

2. NH3 (in low concentrations), and NO3- serve as nutrients
for excessive growth of algae. Plants require, in order of abundance
in plant tissue: C, N, P, and a variety of trace elements. Ex. thick slime
layers on rocks, dense growth of aquatic weeds.
3. The conversion of NH4+ to NO3- consumes large quantities of DO.
Especially where long residence times are available.

(b) In wastewater treatment

1. Biological treatment depends on reproduction of the organisms.

Sufficient N required for the organisms.
2. Domestic wastewater has good C:N:P ratio.
3. In some industrial wastewaters, N is deficient. It must be added.
Phosphorus

(1) Vital nutrient for the growth of algae.

(2) When algae die, they become an O2 – demanding organic material

as bacteria seek to degrade them.

(3) This O2 – demand frequently overtakes DO supply and, as a consequence,

causes fish to die.
 BIOLOGICAL CHARACTERISTICS
• Microorganisms : bacteria, protozoa, worms,
and virus.
• Pathogenic organisms: Common waste water
related diseases- hepatitis A, typhoid, polio,
cholera, and dysentery.
• Major human disease transmission route:
faecal-oral:
– direct(bad personal hygiene)
– indirect ( contaminated food/water)
 Pathogens in wastewater
• Bacteria
– Single celled entities

E.coli
– Size range: 0.5-5 micron
– Consumes soluble food &
capable of self-reproduction
– Diseases: typhoid, paratyphoid,

Polio virus
dysentery, and cholera
• Viruses
– Intracellular parasite
– Size range: ~20-100 nm (approx. 1/50 th of a bacteria)
– Diseases: These are Adenovirus (Respiratory and eye infections),
Poliovirus, Hepatitis A virus, Echovirus (aseptic meningitis), Rota virus
and other virus causing gastroenteritis, diarrhoea.
 Pathogens in wastewater
• Protozoa
–Live attached to the human intestine
where they actively feed and reproduce.
–Common diseases: diarrhea and dysentery.
–Example: Entamoeba histolytica and
Giardia lamblia.
–At some point in their life cycle they undergo a morphological
transformation into a cyst for protection against harsh environment
outside the host. The cyst form is infectious to other persons by the
faecal-oral route of transmission.
–The cysts have size 10-15 micron.

• Helminthes
- Intestinal worms; do not multiply inside human
- Worm burden in infected person is related to
no. of helminthic eggs ingested.
- Size of egg: 40-60 micron; heavier than water
 CENTRAL QUESTION OF BIOLOGICAL
CHARACTERIZATION
• Human carriers exist for all enteric diseases. The fecal-oral route is the
causative pathways in almost all the cases of the disease outbreak
causing public health crisis situations.

• Fecal contamination in wastewater causes the presence of the

pathogens. If not adequately treated, the wastewater and subsequently,
the drinking water shall contain the infectious agents.

• So, in order to be safe, it is regular practice to test water for the

presence of pathogens.

how to assess the pathogenic

• But,
quality when there are so many varieties of
microorganisms in wastewater or water??
 Indicator Organism
•Concept: Rather than testing for each and every pathogen, it is easier to test for only
one group of microorganism whose presence is an assured evidence/ indication that
the wastewater has been polluted by faeces of humans or warm-blooded animals.
This microorganism may be called an indicator organism.
•Indicator organism: Escherichia coli
•Characteristics of E. coli that makes it suitable indicator:
–Non-pathogenic faecal coliform bacteria that reside in the human intestinal tract.
–Excreted in large numbers in faeces, often amounting to about 50 million per gram.
–Untreated domestic sewage contains upwards of 3 million coliforms per 100 mL.
–E. Coli. persists in the environment outside the human intestine for a longer duration
than the other pathogenic bacteria.
–Virus, protozoan cysts and helminth eggs are more persistent than E. Coli.
– But, regular wastewater treatment operations kill all the other pathogens as well as
E. coli. So, E.coli ’s presence is an indicator of presence of pathogens.
–Also, its absence means the faecal contamination is absent.
– It is easier to detect E.coli.
–The severity of faecal contamination is considered to be directly related with the
concentration of the E.coli. bacteria in the water or wastewater.
 E. coli (indicator of faecal
contamination of waste water)
• E. coli colony SEM image of E.coli
Fermentation Tube Technique

Cap Wastewater No growth

Sample and no gas
evolution
Lactose
broth Incubation
Negative
Inverted @ 35 deg C
Vial

Fermentation Tube
Growth with
gas evolution
inside the
inverted vial
Positive
Multiple Tube Fermentation Technique and Most Probable Number

10 mL WW/ or
aliquot

HOW TO STATISTICALLY
X √ √ X √ INTERPRET THE RESULTS??

1 mL WW Statistically found
or aliquot concentrations
are termed as
X X √ X √ Most Probable
Number (MPN) of
the coliform
bacteria present
0.1 mL WW
or aliquot in the wastewater

√ X X X √
Multiple Tube Fermentation Technique and Most Probable Number (cont’d)

Two methods of interpretation of the results of multiple tube fermentation test:

1. Thomas’ empirical method
2. Poisson’s statistical method

Thomas’ Formula:
Most Probable Number (MPN)/ 100 mL =
Number of positive tubes
X 100
mL of samples in negative tubes X mL of samples in all the tubes

Poisson’s statistical method

Maximize y (or ya) by trial and error for different values of λ
1
y = [(1 − e −v1 ) p1 (e −v1 ) q1 ][(1 − e −v2 ) p2 (e −v2 ) q2 ][(1 − e −v3 ) p3 (e −v3 ) q3 ]
a
y = probability λ = Coliform density/ mL vi = sample portion, mL
pi = Number of positive tubes qi = Number of negative tubes a= constant
Derivation of Poisson’s Formula
Consider that a small sample v is taken out of the total wastewater sample volume V.
If there is one single microorganism in volume V.
v
Probability that the small sample contains the microorganism =
V
v
Probability that the small sample does not contain the microorganism = 1 −
V
b
If there are b number of microorganism , probability that  v
the small sample does not contain the microorganism Ps = 1 − 
 V
 vb 
If
v
is very small, then Ps = exp −  S stands for sterile
V
 V 
b
is the density of the microorganism = λ
V
Ps = exp(− v )

So, probability of finding a positive or fertile sample is

Pf = 1 − Ps = [1 − exp(− v )]
In case of multiple number of tubes, if n samples of volume v is taken, the probability of
finding p fertile samples is given by binomial distribution.

𝑛!
𝑦=𝑓 𝑝 = 𝑃𝑓 𝑝 (1 − 𝑃𝑓 )𝑛−𝑝
𝑝! (𝑛 − 𝑝)!
𝑛!
= 𝑃𝑓 𝑝 (1 − 𝑃𝑓 )𝑞 q= number of negative tubes = n-p
𝑝! 𝑞!
𝑛! 𝑝
= [1 − exp −𝑣𝜆 ] [1 − 1 − exp −𝑣𝜆 ]𝑞
𝑝! 𝑞!
𝑛! 𝑝
= [1 − exp −𝑣𝜆 ] [exp −𝑣𝜆 ]𝑞
𝑝! 𝑞!
𝑝! 𝑞!
Denote 𝑎=
𝑛!
1 𝑝
𝑦= [1 − exp −𝑣𝜆 ] [exp −𝑣𝜆 ]𝑞
𝑎
For different dilutions or sample sizes, 1,2, 3 the probability function takes the following form

1
y = [(1 − e −v1 ) p1 (e −v1 ) q1 ][(1 − e −v2 ) p2 (e −v2 ) q2 ][(1 − e −v3 ) p3 (e −v3 ) q3 ]
a