CE 452

ENVİRONMENTAL
ENGİNEERİNG LAB
Sec 1&2

Dr. Al-Ghazawi
BİOCHEMİCAL OXYGEN
DEMAND
BOD
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BIOCHEMICAL OXYGEN DEMAND (BOD)
 BOD the amount of oxygen required by
bacteria while stabilizing decomposable
organic matter under aerobic condition.

Organic matter serve as food for bacteria

Organic
matter

Energy +

oxygen

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 BOD testused to determine pollutional
strenght of domestic and industrial wastes in
terms of oxygen that they will require if
discharged into natural watercources

wastewater
DO
O2
Org.
matter

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 BOD test
◦ Bioassay procedure.
◦ Measures oxygen consumed by living organisms while
utilizing organic matter present in waste.

 Conditions  as similar as possible to those that

occur in nature

 To be quantitative  protect from the air to prevent

reaeration.

 Env. conditions should be suitable for m.o.

 No toxic substance.

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 D.O should be present throughout the test.

O2 solubility~9 mg/L

DO used ~ amount of organic

in sample

Strong wastes must be diluted

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 All necessary nutrients needed for bacterial
growth. (N,P, trace elements)

 Diverse group of m.org. (seed) carry the

oxidation to CO2

 BOD test  a wet oxidation process

 Living organisms serve as the medium for

oxidation of org. matter to water and CO2

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 Oxygen requirement to convert org.
compound to CO2 , water , ammonia.

CnHaObNc + ( n + a/4 - b/2 – 3c/4 ) O2 

nCO2+ (a/2 – 3c/2)H2O+ cNH3

 Temperature effects are held constant by

performing the test at 20C

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 İnfinite time is required for complete biological
oxidation of organic matter

 Practically the reaction considered complete in 20

days =>BOD ultimate

 Large percentage of the total BOD is exerted in

5days

 The test has been developed on the basis of a 5-

day incubation period.=> 5 day BOD

 5 day test was selected also to minimize

interference from oxidation of ammonia.
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BOD Reaction Kinetics
 First order rxn kinetics  Rxn rate proportional to
the amount of oxidizable organic matter remaining
at any time.
(If mo population is nearly constant)

 -dc /dt ˜ C
 -dc/dt = k’C

C: Conc. of oxidizable org matter

K: rate constant

 Rate decreases as C decreases

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In BOD considerations;

 C L (ultimate demand)

 -dL/ dt=k’L

 Lt/L = e-k’t

 Lt=L* e-k’t

 e-k’t = 10–kt

 k=k’/ 2.303

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 k’ : BOD rxn constant (d-1). Typically k’ ˜0.1
to 1 d-1 for degradable org. matter in natural
water at ambient temp 10 < T < 30 °C

BOD t = L (1-10–kt )

BOD t : BOD at any time t,

L: BOD ultimate

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 The oxygen consumed in the time interval
from zero to t is called BOD t

 BOD t : BOD consumed within time t

 Lt : Potential BOD remaining at time t

 BODt = L –Lt = L – L. e-k’t

= L (1- e-k’t )

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 Example 1: Determine 1 day BOD and
ultimate BOD for a wastewater whose 5 day ,
20 °C BOD is 200 mg/ L.

 The reaction constant k ( base e) = 0.23 d-1

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 Its possible to determine reaction constant k
at a temp. other than 20 °C.

 Vant Hoff – Arrhenius relationship :

 Kt = k20 Ɵ ( t-20)

 Ɵ = 1,056 (20<T<30 °C)

 Ɵ = 1,135 ( 4<T<20 °C)

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Example 2: Calculate rate constant for T=24C
k=0.23 d-1 (@ 20C)
Ɵ=1.047

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 Oxygen used  Organic matter oxidized
( Direct Ratio)

~ Oxygen used

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Influence of Nitrification on BOD
 BOD or oxygen-used curve is similar to
organic matter oxidation curve during the
first 8-10 days.

 For proper measurement of BOD, cultures

used in BOD test should contain
heterotrophic bacteria and other organisms
that utilize the carbonaceous matter.

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 Additionally they contain certain autotrophic
bacteria, particularly nitrifying bacteria.
 nitrifying bacteria oxidizes noncarbonaceous matter for
energy

 Nitrifying bacteria present small amounts in

untreated domestic ww.

 @20 °C their populations do not become

sufficiently large to exert oxygen demand until
8-10 days.

 Interference of nitrification eliminated by taking

test period 5 days  BOD 5

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 Untreated domestic ww contains little amount
of nitrifying bacteria, but effluent from
biological treatment units such as Activated
sludge and Trickling filter do contain
nitrifying bacteria that can also consume D.O.
in 5 days.

 Inhibit the action of nitrifying bacteria by

specific inhibiting agents 2-Chloro-6
( trichloromethyl) pyridine (TCMP) or
Allylthiourea (ATU).

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BOD Test

 Method is based on determination of D.O.

Direct method Dilution method

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Direct Method :

If BOD 5 < 7 mg/L  no dilution required.

Ex: River waters

Adjust the sample to 20 °C , aerate with diffused air to

increase or decrease dissolved gas content to near
saturation.

Fill BOD bottles. Measure D.O. İmmediately in first

bottle.

Incubate 5 day  Measure D.O.  BOD =DO1-DO2

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 Dilution Method

Rate at which oxygen is used in dilution of

wastes is in direct ratio to percent of waste in
dilution.

10% dilution uses oxygen at 1/10 the rate of

a 100% sample.

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 Control all environment conditions in the
bioassay test.

 Free of toxic materials

 Favourable pH and osmotic cond.
 Presence of available nutrients
 Standard Temp.
 Presence of mixed organisms of soil origin

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 Industrial wastes may be free of m.org. And
nutrients
 Domestic ww Contain org. Nutrients N, P,
 Dilution water used in BOD must compensate

these deficiences.

Dilution water
Natural surface water
Tap water=>possibility of toxicity from chlorine
residuals.
Synthetic dilution water prepared from distilled,
demineralised w.***the best
Dilution water must be free of toxic subs. Chlorine,
Chloroamines, copper.
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Seeding: (maintain necessary microorganism)
 Domestic wastewaters

Blank (dilution Diluted sample

water)

Seed
Dilution
Nutrient
water
pH buffer

Nitrification inhibitor

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Dilutions of wastes
Set three dilutions

If strenght is known two dilutions

Some casesup to 4 dilutions

 Samples should deplete at least 2 mg/L D.O.

 At least 0.5 mg/L of D.O. remaining at the

end of incubation.
DO1-DO2=2-7 mg/L

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 Example 3: Calculate the percentage of waste
should be added to a BOD bottle if the BOD
range is;

Range, mg/L % sample

200-700 200/2=100
100/100=1 %
40-140 40/2=20
100/20=5 %
20000-70000 20,000/2=10,000
100/10000=0.01%

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Incubation bottles:
 Glass stoppers
 Prevent air trapping during incubation
 Water seal.

 Bottles should be free of organic matter.

 Clean with chromic acid or detergent.
 Rinse carefully
 All cleaning agents removed from bottle

◦ 4 rinse with tap water,

◦ final rinse with Distilled or deionized water

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Calculation of BOD

𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑏𝑜𝑡𝑡𝑙𝑒
൤ሺ𝐷𝑂𝑏−𝐷𝑂𝑖ሻ ൨− ሺ𝐷𝑂𝑏−𝐷𝑂𝑠ሻ
BOD(mg/L)= 𝑚𝑙 𝑠𝑎𝑚𝑝𝑙𝑒

DOb=DO found in blank at the end of incubation

DOi=DO found in diluted sample at the end of incubation
DOs= DO that originally present in undiluted sample

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 Example 4: Calculate BOD concentration in
mg/L

DOb 7
V sample 2
V bottle 300
DOi 2
DOs 4

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Respirometric measurement

OXYTOP

Respirometric method:Respirometric
methods provide direct measurement
of the oxygen consumed by
microorganisms from an air or
oxygen-enriched environment in a
closed vessel under conditions
of constant temperature and agitation.

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Rate of Biochemical Oxidations
Differs in diff. Wastes

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 Effluent from bioligical wwtp different from
raw wastes
◦ Nature of the org. Matter
◦ The ability of the organisms present to utilize the
org. Matter
◦ Rate of hydrolysis and diffusion

 Ex: Glucose high k

 Lignin, Synthetic detergents

 slowly attacked by bacteria

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BODL and Theoretical Oxygen Demand

 Total BOD or L = ThOD considered to be

equal

 Oxidation of glucose

C6H12O6 + 6O2  6CO2+ 6H2O

180 192

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192 g O2 /mole of glucose
OR
1.065 mg O2 /mg glucose

300 mg/L of glucose ThOD=

300*192/180=320 mg/L

Experimentally;
BOD measurements (20 day)
BOD(L)= 250-285 mg/L
85% of theoretical amount

Not all the glucose converted to CO2 and water

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 Org. Matter  Food material 
◦ Energy
◦ Growth
◦ Reproduction

Part of org. Matter  Converted to cell tissue

Cell tissue will remain unoxidizedtill endogeneous

respiration

◦ When bacteria die they become food material for others.

◦ Further transformation to CO2 , H2O and cell tissue

◦ Living bacteria + Dead ones  Food for higher organisms 

Protozoans
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 A certain amount of organic matter remains
in these transformations.

 Resistant to further biological attack.

 Humus amount of org. matter

corresponding to discrepancy between total
BOD and ThOD.

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Analysis of BOD Data

Calculation of k value is needed to obtain L using

BOD5.
k and L are determined from a series of BOD
measurements.

Methods:
 Least squares

 Thomas Method

 Methods of moments
 Daily-difference method

 Rapid ratio method

 Fujimoto Method

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Least squares method:

Fitting a curve throug a set of data points. Sum

of the squares of the residual must be
minimum. (Difference between observed and
the fitted value)

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 dy/dt (t=n) = k (L-yn )

 For the time series of BOD measurements on

the same sample Eqn. May be written for each
of the various n data points.

 k and L  unknown

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 If it is assumed dy/dt represent the slope of
the curve to be fitted through all data points
for a given k and L value, two sides will not
be equal because of experimental error.

Difference  R
R=k(L-y)-dy/dt
R=kL-ky-y’
a=kL -b=k
R=a+by-y’

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