10 Filtration - Merged

Chair of Urban Water Systems Engineering
Technical University of Munich
Filtration
Prof. Dr.-Ing. Jörg E. Drewes

30 November 2023
Slides with background information, review on your own
2
3
Flocculation/Sedimentation
4
Overview
1. Introduction
2. Removal Mechanisms
3. Classification of Filters
4. Filter Media
5. Filter Components
6. Hydraulics
7. Rapid-Filter Design
5
1. Introduction to Filtration
• Removal of particulate matter from water including algae, sediments, clay,
organic and inorganic particulate, microorganisms
• Nearly all surface water treatment facilities use filtration
• Historically:
– Has been used to clarify water for thousands of years
– First modern slow sand filtration in London (1852)
– Interest in filtration grew as people realized it can prevent waterborne diseases
– Rapid filtration originated in the U.S. during the 1880s
6
Cholera Outbreak Hamburg 1892

• Drinking water abstraction from the
river Elbe
• No filtration
• 9,000 deaths
• Very limited impacts in Altona
7
William Philipps Dunbar
…a pioneer in
public health,
bacteriology,
allergology, and
sanitation engineering
*27.10.1863, +19.03.1922
www.wikipedia.com
Relevant Regulations
• European Union (2020/2184) – Drinking Water Directive
– Turbidity for surface water treatment should be below 0.3 NTU and
never exceed 1 NTU in the finished water
• Trinkwasserverordnung (TrinkwV) 2023

– Turbidity for finished water should be below 0.3 NTU and never exceed
1.0 NTU in the finished water
• U.S. EPA - Surface Water Treatment Rule

– As of January 1, 2002, for systems servicing >10,000, and January 14,
2005, for systems servicing <10,000, turbidity may never exceed 1 NTU,
and must not exceed 0.3 NTU in 95% of daily samples in any month
9
Common Filtration Processes

• Three common types:
– Rapid filtration – most common in Europe and the United States
– Slow sand filtration – small utilities / high quality feed water
– Pre-coated filtration – specific applications (green sand filtration)
10
Comparison between Granular Filtration

Technologies for Water Treatment
11
Rapid Granular Media Filtration

• Replaced slow sand filtration
• Up to 100 times faster due to:
– Granular media processed to have uniform size
– Coagulation is used for pretreatment
– Mechanical and hydraulic systems to efficiently remove solids from
the bed
12
Rapid Filtration – Process Flow

• Filter bed is contained in a deep structure (concrete, open)
• Two-stage cycles:
– Filtration cycle
• water flows downward through the filter
• particles accumulate
• typically lasts 1-4 days
– Backwash cycle – accumulated material is flushed from the system

• water flows upward
• efficient removal of solids is a key component of rapid filtration
• typically lasts 15-30 minutes
13
14
https://www.youtube.com/watch?v=UmzR3TRTz2U 15
2. Mechanism of Filtration
• Physical treatment process to remove colloidal particles
• Removal mechanisms during filtration

– Straining (i.e., removal due to size)
– Sedimentation (i.e., particles in low-velocity zones)
– Impaction (i.e., particle removal due to inertia)
– Interception (i.e., particle contact with media grain)
– Diffusion (i.e., removal due to Brownian motion)
16
Straining
• Capture of spherical particle by spherical media
grains
• If the ratio of particle diameter to media

diameter is greater than 0.15, the particle will be
strained by the media
• If the ratio is smaller, straining is not possible and

particle capture occurs by other means
• For typical rapid filtration, straining is limited to

particles with diameter of 80 µm and larger
17
Sedimentation
• Particles in low-velocity zones
• Particles with a density significantly greater

than water tend to deviate due to
gravitational forces
18
Impaction
• Particles with an inertia greater than the
hydrodynamic force will tend to deviate
• Insignificant in water treatment!
19
Interception
• Particles that pass the collector surface by a

distance of half the particle diameter will
contact the collector and be intercepted
20
Diffusion
• Particles influenced by Brownian motion
will deviate from the fluid streamlines
due to diffusion
• Diffusion is most significant for particles

< 1 µm in diameter
21
Removal Mechanisms During Filtration

Particle transport mechanisms in fundamental
filtration theory:
1. Interception: particle A follows a streamline but
collides with the collector
2. Inertia, sedimentation: particle B deviates from the

streamline and collides with the collector because of
inertial or gravitational forces
3. Diffusion: particle C collides with collector due to

random Brownian motion
particle diameter < 80 µm

22
Attachment efficiency
• Transport efficiency depends on particle
size and density
– Small particles removed by diffusion
– Larger particles removed by
sedimentation and interception
• Lowest efficiency for particles of 1-2 µm
23
Principal Attraction Forces
• Chemical Bonding
• Physical Attraction
– Electrostatic forces
– van der Waals forces
24
Filtration Dynamic
• As filtration continues, pore spaces between media grains
become smaller
– enhance straining
– increased velocity of water through the pore spaces
– increased shear forces of the water flowing through the media bed
– increased head loss
I decrease in filtration rate, filter performance, and

efficiency
25
3. Classification of Filters
• Filtration rate
– slow sand filters
– rapid filters
– high-rate filters
• Driving force
– gravity
– pressure (up to 1 MPa (10,3 bar))
• Direction of flow
– downflow – most commonly used in water treatment
– upflow – rarely used in granular filters
• Pretreatment level
26
Slow Sand Filter

• Hydraulic loading rate < 10 m3/m2-day
• Does not utilize chemical coagulation
• Removal occurs in the upper layers of the filter
27
Slow Sand Filter
28
Slow Sand Filter
• River bank filtration (RBF)
29
Rapid and High-Rate Sand Filters
• Hydraulic loading rate ~120 m3/m2·day (rapid)
• Hydraulic loading rate >240 m3/m2·day (high-rate)
• Designed for removals throughout the entire depth of

the filter bed
• Most of these filters have multiple layers
30
Filters Classified by Driving Force

Gravity filters: Pressure filters:
• 2-3 m head • higher head
• housed in open concrete • housed in closed steel vessels
or steel tanks • costly; small systems
• large and small systems
31
Rapid Filtration – Pretreatment

• Coagulation is required for high removal efficiency
• Destabilization of particles will reduce repulsion forces between

the filter media and the particles
32
Classification of Rapid Filtration

Systems by Pretreatment Level
Conventional filtration
Direct filtration
In-line filtration
Two-stage filtration
33

34
4. Filter Media
• Media characterization
– Effective Size
– Uniformity coefficient
• Materials for Rapid-Filtration
• Filter classification
– single-medium
– dual media
– mixed media
35
Media design parameters

• Size distribution determined by sieve analysis
• effective size (d10)
𝐸𝑆 = 𝑑!"
• uniformity coefficient
𝑑#"
𝑈𝐶 =
𝑑!"
36
Example 1: Determine ES and UC

• Determine effective size and uniformity coefficient of
granular materials
37
Stratification/reverse gradation
38
Rapid Filtration – Typical Media

Properties
39
Sand
• worn or crushed sand
0.4 – 0.8 mm
40
Anthracite
0.5 – 2 mm
41
Garnet
0.3 – 0.8 mm
42
Filter classification
43
Single-Medium Filter
• stratification/reverse gradation
44
Ideal Filter
45
Dual-Media Filter
• Anthracite coal (1.55) and quartz sand (2.65)
• More depth is utilized
46
Dual-Media Filter
47
Mixed-Media Filter
• anthracite coal (1.55), quartz sand (2.65), garnet (4.05)
48
Filter Media
• Typical Media Design Values
49
Filter Media Selection
Kawamura relationships 50
5. Filter Components
51
Filter Support Media and Underdrains
52
Filter Components
53
Filter Components
54
Filter Components
Filter Backwash 55
6. Hydraulics of Flow Through Granular

Media
• Flow regime in granular media identified
by Reynolds number for flow around
spheres: r vd
Re = W
µ
rW = fluid density, kg/m3
v = filtration rate (superficial velocity), m/s
d = media grain diameter, m
µ = dynamic viscosity, kg/m·s
56
Hydraulics of Flow Through Granular Media
• Steady, laminar flow

– Re < 1 : Darcy flow or creeping flow – viscous flow
behavior (HL=f(v))
– 1 < Re < 100 : Forchheimer flow – viscous and
inertial (HL=f(v2)) behavior
• 100 < Re < 600-800 : Transition flow
• Re > 800 : Turbulent flow

57
Ranges of Reynolds Number in Granular

Filtration Technologies
58
Headloss Through Clean Granular Filters

• Net available head = filter design headloss –
clean-bed headloss
• Insignificant in slow sand and pre-coat

filtration, but important in rapid filtration
59

• Henry Darcy (1856) – relationship between
velocity, hL, and bed depth in granular media
under creeping-flow conditions:
kp = hydraulic permeability, m/s
hL v = filtration rate (superficial velocity), m/s
v = kp hL = headloss across media, m
L L = depth of granular media, m
• Not taking into account media properties…

• Poiseuille’s Law and Kozeny development:
hL 32 µv k k µS 2v
= =
L rW gd 2
rW ge 3
60

• Forchheimer flow regime – nonlinear relationship for viscous and
inertial effects:
hL k1 = permeability coefficient (viscous), s/m
= k1v + k 2v 2 k2 = permeability coefficient (inertial), s2/m2
L
• Later development with correlations to measurable properties:
hL = kV
(1 - e )2 µLv 1 - e Lv 2
+kI 3
Ergun
Equation
e3 rW gd 2
e gd
kV = headloss coefficient (viscous), unitless
kI = headloss coefficient (inertial), unitless
61

• Effects of porosity and velocity
62
Example 2
Calculate the clean-bed headloss through a deep-bed filter with 1.5 m
anthracite coal (ES=1.1 mm) over 0.3 m sand (ES=0.5 mm) at a
filtration rate of 15 m/hr and temperature of 15 °C
Ø rW = 999 kg/m3, µ = 1.14·10-3 kg/m· s

Ø midpoints from Table 11-5:
anthracite coal: kV = 228, kI = 4.4, e = 0.50
sand: kV = 112, kI = 2.2, e = 0.42
hL = kV
(1 - e )2 µLv 1 - e Lv 2
+ kI 3
e3 rW gd 2
e gd
63
Example 2
64
Backwash Hydraulics
• Backwash flowrate must be great enough to flush

captured material from bed
• …but not to the point that filter media is flushed out of
the filter box…
• To prevent loss of media, need determine Bed Expansion
65
Rapid Filtration – Backwashing

1. Filter influent and effluent lines closed
2. Backwash supply and waste wash water valves opened
3. Backwash water (potable water) is directed upward
4. Backwash supplemental systems:
– Surface wash systems – nozzles blast surface of bed
– Pressurized air under the
media – reduces water usage
5. Valves positions reversed
and filter is placed back
in service
66
Forces on Particles (foulants or grain)

• Backwashing hydraulics occur within the Forchheimer flow
regime
æp ö
• Gravity: FG [N ] = mg = (rP - rW )ç d 3 ÷g
è6 ø
• Upward drag forces: FD [N ] = 3pµvd for Re < 2

2.31
FD [N ] =
0.6
prW v 2 d 2 for 2 £ Re £ 500
Re
• Equating the two allow to determine the fate of individual
particles
é g (r P - rW )d 1.6 ù
0.714
v=ê 0.4 0.6 ú

ë 13 . 9 rW µ û
67
Example 3
• A filter is backwashed at 40 m/hr at 15°C.

Determine whether a 0.1 mm particle of sand will
be washed from the filter.
68
Example 3
69
Bed Expansion and Porosity

• The flow velocity in fixed bed is higher than for isolated particle
because of the presence of media à higher drag forces
• As media rises, porosity increases and velocity decreases until
equilibrium is reached
• Relationship between bed porosity and expansion…
𝐿$ 1 − 𝜀% LE
= LF
𝐿% 1 − 𝜀$
70
Multi-media Filters
• The media in multi-media filters must be
matched so that all media fluidize at the same
backwash rate
é g (r P - rW )d 1.6 ù
0.714
v=ê 0.4 0.6 ú

ë 13.9 rW µ û
• Equating an equivalent fluidization velocity for

two media and solving for grain size ratio yield
the expression:
0.625 d1 = grain diameter of one filter medium, m
d1 æ r 2 - rW ö d2 = grain diameter of a second filter medium, m
= çç ÷÷ r1 = density of medium with diameter d1, kg/m3
d 2 è r1 - rW ø r2 = density of medium with diameter d2, kg/m3
71
7. Design of Rapid-Filters
Preliminary design consists of several tasks:
1. Setting performance criteria
n effluent turbidity n length of filter run
n recovery n unit filter run volume (UFRV)
2. Selecting process design criteria

n required level of pretreatment n filter media type, size, and
depth
n filtration rate n available head
3. Selecting method for flow distribution and control
4. Selecting major process components (backwashing systems, underdraines,

wash troughs, process piping)
72
Rapid Filtration – Filter Run

Effluent turbidity characterized by three distinct
phases:
1. Ripening
– media conditioning (15 min - 2 hr)
– Sometimes consists of two peaks
– Size and duration can be reduced by
proper backwashing procedure
– Filter-to-waste line
2. Effective filtration
– Steady state with turbidity <0.1 NTU
– Gradual increasing in head loss
3. Breakthrough
– Filter bed can’t hold more particles
– Effluent turbidity increases
– Headloss increases
73
Rapid Filtration – End of Filter Run

Can be triggered by several events and lead to backwash:
1. Breakthrough
n Preventing high-turbidity water from entering
the distribution system
2. Headloss
n Increases beyond the available head through
the process
n Rapid filters are typically designed with
1.8 - 3 m available head
n If in specific cases neither are reached
within several days: utility initiates
backwash after a set period of time
74
Performance Criteria: Effluent Water Quality

• Turbidity is used as a surrogate for the concentration of particulates in water
• Current US regulation: “…As of January 1, 2002, for systems servicing >10,000,

and January 14, 2005, for systems servicing <10,000, turbidity may never
exceed 1 NTU, and must not exceed 0.3 NTU in 95% of daily samples in any
month”.
• Europe/Germany: “…less than 1.0 NTU”
• Typical goal: 0.1 NTU (close to detection limit)
• Particle counter is a newer method for measuring effluent quality (provides

both number and size distribution of particles in water)
75
Performance Criteria: Length of Filter Run
• Dictates how often backwash is performed
• Affects water recovery and how much labor involved
• Minimum desirable: 1 day
• Common design length: 1-4 days
76
Performance Criteria: Recovery

• Ratio between the net and the total quantity of water filtered
• Some product water is used for backwash and some is discharged as filter-to-
waste
• Recovery is evaluated using the concept of

– Unit Filter Run Volume (UFRV)
– Unit Backwash Volume (UBWV), and
– Unit Filter-to-Waste Volume (UFWV)
77
Recovery
a = filter cross-sectional area, m2
VF VF = volume of water filtered during one filter run, m3
UFRV = = v F tF
a VBW = volume of water for backwashing one filter, m3
VFTW = volume of water discharged as filter-to-waste, m3
VBW
UBWV = = v BW t BW vF = filtration rate (superficial velocity), m/hr (=vFTW)
a vBW = backwashing rate, m/hr
VFTW tF = duration of filter run, hr
UFWV = = v F t FTW
a tBW = duration of backwash cycle, hr
tFTW = duration of filter-to-waste period, hr
VF - VBW - VFTW UFRV - UBWV - UFWV

Re cov ery º r = =
VF UFRV
78
Recovery
• Effective filtration rate (vEFF) º net volume produced over total
time
UFRV - UBWV
vEFF =
t F + t BW
• Filters should be designed to achieve recovery of at least 95%
• Typical wash water quantities are ~8 m3/m2 à to achieve 95%,

UFRV of at least 200 m3/m2 is required!
79
Relationship between Recovery, UFRV and UBWV
80
Example 4
• A filter is operated at a rate of 10 m3/m2-hr for 37.5 hrs, of which 15
minutes was discharged as filter-to-waste. After filtration, the filter is
backwashed at a rate of 40 m/hr for 15 minutes.
• Calculate: UFRV, UBWV, and UFWV, effective filtration rate, and recovery
81
Example 4
82
Process Design Criteria

• Filter type
• Filtration rate
• Number and Dimensions of Filters
• Available Head
• Filter Media
83

Conventional filtration
Direct filtration
In-line filtration
Two-stage filtration
84
Filtration Rate
• Filtration rate influences many aspects of rapid-filter design:
– Clean-bed headloss
– Rate of headloss accumulation
– Distribution of solids collection in bed
– Effluent quality
– Run length
• Also directly related to cost
• Highest filtration rate that yields good filter performance is

recommended
85
Filtration Rate
• Filtering floc resulting from alum or ferric with polymer aid, up to 25 m/hr
• Filtering weak floc of alum without polymer aid, <12.5 m/hr
• Often subjected to regulatory limits (U.S. west coast states <15 m/hr)
• Most rapid filters are designed to operate at 5 – 15 m/hr
86
Number & Dimensions of Filters

• Number of filters is influenced by the
– capacity of the plant
– dimensions of a single filter
– effect of filtration rate changes during backwashing
– economic considerations
• Most water treatments have a minimum of 4 filters
• Filters are typically designed to treat the maximum plant capacity at the
design filtration rate with at least one filter out of service
87
Number & Dimensions of Filters

A (total filter area) = Q (peak day) / v (selected)
• Maximum dimensions of a single filter are generally determined by

the economic size of backwash facility and difficulties in providing
uniform distribution of backwash water
• Typical maximum size of one high-rate gravity filter is ~100 m2
• Extremely large plants sometimes have double this size
88
Available Head
• Filtration head = available head in the structure – clean-bed headloss
• Rapid filters typically operate by gravity
• Available head depends on upstream and downstream structures
• Selection of available head involves trade-off between longer filter

runs (greater available head) and economics (smaller available head)
• Due to construction costs, available head is rarely more than 2 – 3 m

89
Backwashing Systems Typical Design Criteria
90
Design Criteria for Filters at Typical WTP
91
Control of Filtration Rate

• Modulating control valve:
– Flow meter controls a modulating control valve on the effluent piping
from each filter
– Set-point for total flow
– Advantages: flexibility, smooth flow of influent without weirs or orifices
– Disadvantages: complex control system, high maintenance, can cause

variation in filtration rate
92

• Influent weir flow splitting:
– Water flows through a common influent channel to several filters
– Influent weir for each filter
– When filter is removed from service for backwash, flow automatically
distributed to the remaining filters
– The simplest method
– Rate changes are made gradually with no control valves
93
Hydraulic Modes
• Constant Rate and Constant Head Water Filtration:
– Most common filtration mode
– Water head in the filter box is at highest level throughout the duration of
filter run
– A valve on the effluent line controls filtration rate
• beginning of run – valve almost completely closed
• end of run – valve almost completely open
94
Hydraulic Modes
• Constant Rate and Rising Head Water Filtration:
– Effluent valve remains open throughout the filter run
– The head water seeks its own level
– Head water should be deep enough to prevent scouring of top layer
when water spills over the overflow launders
– Water head in the filter box rises to its terminal level
– No need for sophisticated control
95
Hydraulic Modes
• Declining-rate filtration:
– Effluent valve remains open throughout the filter run
– All filters receive water from a common influent channel
– All filters operate at the same head
– Each filter receive a different quantity of influent water based on the
accumulated solids (headloss) in the filter
– The flow through each filter declines as solid accumulate
– The common water level gradually rises as all filters accumulate solids
– Backwash is initiated on a run length basis
96

• Declining-rate filtration (cont’):
– Advantages:
• simplicity, no instrumentation or flow control on individual filter
– Disadvantages:
• no indication of flow rate or headloss in any individual filter
• no method to control filtration rate
• the initial filtration rate can exceed the designed rate
• Discouraged by many state regulating agencies
97
Take Home Messages

• Filtration principles
• Mechanisms of filtration; attachment theory
• Classification of filtration types
• Media design
• Filter components
• Head loss calculations

98
Wastewater Characteristics I

7 December 2023

1
Wastewater Characteristics
- Domestic sewage contains approximately 99.9% water!
- The remaining portion includes organic and inorganic matter, as well as suspended and
dissolved solids (incl. microorganisms)
water
treatment solids
pollution
because of this 0.1% that water pollution occurs and wastewater

needs to be treated prior to discharge to the environment
adapted from von Sperling (2005) 2

Wastewater Characteristics
Origin of wastewater – input
material flux analysis rain water
settlement
input output
- water - wastewater
- food Suspended solids;
-chemicals organic pollutants;
-consumer goods, etc. inorganic pollutants;
heavy metals;
bacteria;
viruses
output …
(roof, courtyard and road runoff)
Transformation Processes in the Environment
Input
Growth:
• Plants
Sedimentation
• Algae
Erosion
• Fish
input • Protozoa
• Bacteria
Advective transport Self purification
-Nitrification,
Flow -Denitrification,
Diffuse inputs -transformation of
organic matter, … 4
Introduction to Biological Treatment

However, in natural water bodies: Self purification is slow
big reactor volumes (ponds)
Often more advantageous: Optimization of reaction rates
Increasing biomass in a reactor
Increasing oxygen
Enabling optimal contact between biomass, pollutants and oxygen
5

Objectives of (traditional) wastewater treatment:
• Protection of the receiving waters with minimal costs
• Transformation into substances that have no or little damaging effect on the receiving
system or can be separated from the wastewater effluent
River
Solution:
Transforming the natural conversion
processes into engineered systems
Biofilm reactor Reactor with biomass in suspension

Source: Mudrack und Kunst, 2003 6
Municipal Wastewater Treatment Plants
Source: Marcus Östman; CC0 Public Domain 7

Conventional Activated Sludge (CAS) Treatment
Source: Marcus Östman 8


• In biological wastewater treatment the transformation is achieved by
microorganisms
• The microorganisms used in the system are a

complex mix of different types of
microorganisms with (very) different
characteristics regarding their activity (function)
• The composition and properties of this complex mixed biomass depends on

the conditions characterized by the technological system and the
characteristics of the wastewater and is subject to dynamic processes
9
Dominant Bacterial phyla and domain Archaea

of Activated Sludge
Vuono et al. (2016), ES&T


• The task of biological wastewater process engineering is to design the system in a way
that the conditions within the system promote the stable growth of those
microorganisms that have the desired properties/function
• Desired properties include:

- Organisms being able to degrade / transform specific undesireable wastewater
constituents at a reasonable rate
- Organisms must easily be separated from the

effluent (e.g. easily settleable)
+++++++
+++++++++ +++++
+++++++ ++++++
+++
++
+
++++
++
+++
++
+
+
+++++++++
+++++
++++ ++++
++
+++
+
+
++++
++
+
+
+++
+++++++
++
+
+++
++
11

Fundamental processes:
Solid-liquid separation suspension clarified effluent
concentrate
Reaction unit; educts products

biochemical transformation
reactor
12

Example activated sludge system:
• Co-existense of aerobic, anoxic, and anaerobic bacteria
Objectives:
Elimination of organic matter (COD), elimination of N (Nitrification, Denitrification),
and phosphorus (Bio-P-Elimination)
NO3
COD
Denitrification Nitrifikation
Creation of alternating conditions for the microorganisms in the system in order to

control the composition and transformations of the types of organisms present
13
Changes in effluent
quality over time
14
Fundamental Transformation Processes
Degradation of organic carbon (aerated à aerobic conditions)

e.g. methanol: 2 CH3OH + 3 O2 è 2 CO2 + 4 H2O
Heterotrophic bacteria
Nitrification (aerated à aerobic conditions)

2 NH4+ + 3 O2 è 2 NO2- + 2 H2O + 4 H+ (nitritation)
2 NO2- + O2 è 2 NO3- (nitratation)
Autotrophic bacteria
Denitrification (not aerated à anoxic conditions)

5 CH3OH + 6 NO3- è 5 CO2 + 7 H2O + 3 N2 + 6 OH-
15
Transformation of N-species during Biological Treatment
Source: Weißbach, M. Criddle, C.S., Drewes, J.E.., Koch, K. (2016). Environmental Science: Water Research and Technology, 3 (1), 10–17. 16

Importance of wastewater characteristics
• Wastewater characteristics influence the microorganisms and microbiological

processes
• Thus, wastewater characteristics determine the optimal design of the system and
are essential for predicting the performance of the system
• To design an (activated sludge) treatment process properly, characterization of the

wastewater is perhaps the most critical step in the process
• Without comprehensive wastewater characterization, facilities may either be

under- or overdesigned, resulting in inadequate or inefficient treatment
17
Sink 3
gases
effluent
influent
Sources WWTP Sink 1
waste sludge
Sink 2
18
Bulk, Group or Lumped Parameters
19
Lumped Parameter
• wastewater is a complex mixture of different compounds that are only partially known
• There is a vast amount of different biochemical and microbiological processes that are carried
out in a reactor in biological wastewater treatment
• However,
- typically many fundamental aspects of the processes and their performance are characterized by
the behaviour of substances, that can be described by means of lumped parameters (e.g., oxygen
consumption, sludge production)
- considerung the day-to-day operations at a wastewater treatment plant, usually there are no
resources for a detailed analysis of single substances
20
Lumped Parameters
• The „backbone“ of biological wastewater treatment processes can be described by

means of ‚lumped parameters‘
• Lumped parameters represent a group of single substances that are characteristized

by a specific common property. The property chosen depends on the problem to be
solved
• Thus, lumped parameters represent only a specific aspect of the sum of the
substances
• If a statement regarding single substances is required, these lumped parameter can

serve as a basis for a more detailed description of single substances
21
Lumped Parameters
An illustrative example
All wastewater
constituents
Lumped parameter: Single substance:

a subset one element
Set of Set of
particulate dissolved
substances substances
Separation, e.g., membrane filtration

Gujer, 1999 22
Main classification of constituents
23
Main Classification of Constituents

Characterization by size,
Characterization by settleability
chemical characterisitics
• non-
settleable
• inorganic
compounds •
settleable
• organic • dissolved
compounds (soluble)
• colloidal
• suspended
(particulate)
Characterization by
biological degradability
• biodegradable
• non
biodegradable
24

Physical Characteristics
solid
s
temperature
Metcalf & Eddy 25


Inorganic Chemical Characteristics
nitrogen,
phosphorus
Metcalf & Eddy 26


Organic Chemical Characteristics
carbonaceous matter,
oxygen demand
Metcalf & Eddy 27

Representation of Wastewater Constituents

Chemical Characterization Physical Characterization
- - size
volatilization
- settleability
Biological Characterization
- biodegradability
28
Solids
29
Solids
• All the contaminants in water, with the exception of dissolved gases, contribute to the solids load
Suspended solids coarse material i.e., plastic bags, sanitary napkins

settleable material i.e., sand, grit
suspended matter i.e., bacteria
floating matter i.e., fats / oil
Why treating? development of sludge deposits and anaerobic conditions in

aquatic environment
30
Solids
Classification of solids
Solids
Classification by size Classification by Classification by

chemical settleability
characteristics
Volatile Fixed
(organic) (inorganic)
Dissolved colloidal Suspended
(soluble) (particulate settleable non-
) settleable
31
Solids
Classification by size SS: dry solid on filter at 105 oC
TDS: dry filtrate at 105 oC

Dissolved solids (soluble solids):
• particles of small dimensions
• capable of passing through a filter paper of a specific size
(dissolved solids = filterable solids)
Suspended solids (particulate solids):

• particles with larger dimensions
• retained by the filter (suspended solids = non-filterable solids).
colloidal solids:
• in an intermediate range
• of importance in water treatment, but
• difficult to identify by the simple method of paper filtration.
• the major part of colloidal solids is separated as filterable
(dissolved) solids.
Total solids = suspended solids + dissolved solids
32
Solids
Basic procedure for classification by size Wastewate
r sample
Filter
filtrate solids on filter

105°C
evaporation evaporation
dissolved suspended
solids solids
• Filterable • Non-filterable
Total solids = suspended solids + dissolved solids
33
Solids
Particle Size Ranges
algae
bacteria
Bacterial flocs
cell fragments
viruses
protein
DNA Visible to the naked eye
fatty acids
ammonium,
nitrate, etc..
settleable
dissolved colloidal
suspended
10-6 10-5 10-4 10-3 10-2 10-1 100 10+1 10+2 µm

1 µm=0.001 mm
34
Solids
Basic procedure for classification by organic Wastewater
matter sample
105°C
evaporation
Total solids
550°C
Volatile
solids
(volatilized, Fixed solids
oxidized) (inert fraction,
• Representation of unoxidised)
organic matter
• Representation of inorganic
Total solids = Volatile solids + Fixed solids or mineral matter
35
Solids
Classification by Settleability
Standard test for settleable solids:
• Placing a wastewater sample in a 1-liter Imhoff cone
• Noting the volume of solids in millimeters that settle after a specified time period.
• The volume of solids accumulated in the bottom of the Imhoff Cone is measured and expressed as mL/L
Settleable solids:
are considered those that are able to settle in a specified period.
(typically approx. 60%)
Non-settleable solids:
The fraction that does not settle represents the non-settleable
solids.
Total solids = settleable solids + non settleable
36
Solids
Classification I TOTAL SOLIDS (TS)
The residue remaining after a wastewater
sample has been evaporated and dried at a
specified temperature (103 to 105°C)
TOTAL TOTAL DISSOLVED

SUSPENDED „Abfiltrierbare SOLIDS (TDS)
SOLIDS (TSS) Stoffe“ Those solids that can pass through the
filter, and are then evaporated and dried
Portion of the TS retained on a filter
at a specified temperature. Consists also
with a specified pore size,
of colloidal besides the “truely” dissolved
measured after being dried at a
solids!
specified temperature (105°C)
VOLATILE FIXED TOTAL VOLATILE FIXED

SUSPENDED SUSPENDED DISSOLVED DISSOLVED
SOLIDS (VSS) SOLIDS (FSS) SOLIDS (VDS) SOLIDS (FDS)
Those solids that can be The residue that Those solids that can be The residue that remains
volatilized and burned off remains after TSS are volatilized and burned off when TDS are ignited
when the TSS are ignited (500 ignited (500 +/- 50°C) when TDS are ignited (500 +/- (500 +/- 50°C)
+/- 50°C) 50°C)
„Glühverlust der „Glührückstand der

abfiltrierbaren Stoffe“ abfiltrierbaren Stoffe“
37
Solids
TOTAL SOLIDS (TS)
Classification II The residue remaining after a
wastewater sample has been
evaporated and dried at a specified
temperature (103 to 105°C)
TOTAL VOLATILE SOLIDS TOTAL FIXED SOLIDS (TFS)

(TVS) The residue that remains after TS are
ignited (500 +/- 50°C)
Those solids that can be volatilized and
burned off when the TS are ignited (500
+/- 50°C)
VOLATILE TOTAL VOLATILE

FIXED FIXED
SUSPENDED DISSOLVED
SUSPENDED DISSOLVED
SOLIDS SOLIDS (VDS)
SOLIDS (FSS) SOLIDS (FDS)
(VSS) Those solids that can be
The residue that remains The residue that remains
Those solids that can volatilized and burned off
after TSS are ignited (500 when TDS are ignited
be volatilized and when TDS are ignited (500
+/- 50°C) (500 +/- 50°C)
burned off when the +/- 50°C)
TSS are ignited (500
+/- 50°C)
„Glührückstand der
„Glühverlust der abfiltrierbaren Stoffe“
abfiltrierbaren Stoffe“
38
Solids
Overlapping of Classification
Approaches
39
Solids
Example 1
The following test results were obtained for a wastewater sample taken at the headworks to a wastewater
treatment plant. All of the tests were performed using a sample size of 50 mL. Determine the concentration of
-Total solids (TS),
-Total volatile solids (TVS),
-Suspended solids (TSS),
-Volatile suspended solids (VSS),
-Total dissolved solids (TDS)
-Total volatile dissolved solids (VDS).
The samples used in the solids analysis were all either evaporated, dried, or ignited to constant weights.
-Tare mass of evaporating dish = 53.5433 g
-Mass of evaporating dish plus residue after evapor. at 105°C=53.5794 g
-Mass of evaporating dish plus residue after ignition at 550°C = 53.5625g
-Tare mass of filter after drying at 105°C = 1.5433 g
-Mass of filter and residue after drying at 105°C = 1.5554 g
-Mass of filter and residue after ignition at 550°C = 1.5476 g
40
Solids
Solution to example 1
Given: -Tare mass of evaporating dish = 53.5433 g
Total solids (TS) = residue after evaporation at 105°C / 0.05 l
= (tare mass of evaporating dish + residue – tare mass of evaporating dish ) / 0.05 l
53.5794 g – 53.5433 g = 0.0361 g
à TS = 0.0361 g / 0.05 l = 0.722 g / l = 722 mg/l
Total volatile solids (TVS) = TS – TFS
TFS = (residue after ignition at 550°C ) / 0.05 l
TVS = TS - (tare mass of evaporating dish + residue after ignition at 550°C ) / 0.05 l + (tare mass of
evaporating dish) / 0.05 l
TVS = 722 mg/l - 53562.5 mg / 0.05 l + 53543.3 mg/ 0.05 l
à TVS = 338 mg/l
41
Solids
Calculated: TS, TVS
Total suspended solids (TSS) = residue on filter after filtering / 0.05 l
= (tare mass of filter + mass of residue)/0.05 l – (tare mass of filter) / 0.05 l
= 1.5554 g / 0.05 l - 1.5433 g / 0.05 l
à Total suspended solids = 0.0361 g / 0.05 l = 242 mg/l
Volatile suspended solids (VSS) = TSS – FSS

FSS = The residue (of what stayed on filter) that remains after TSS are ignited / 0.05 l
VSS = TSS + tare mass of filter / 0.05 l - (residue on filter after ignition + tare mass of filter) / 0.05 l
= 242 mg/l + 1543.3 / 0.05 l - 1547.6 mg / 0.05 l
à Volatile suspended solids = 156 mg/l
42
Solids
Calculated: TS, TVS, TSS, VSS
Total dissolved solids (TDS) = TS – TSS

Total dissolved solids = 722 mg/l - 242 mg/l = 480 mg/l
Volatile dissolved solids (VDS) = TVS – VSS

àVDS = 338 mg/ l – 156 mg/l = 182 mg/l
43
Solids
+tare mass of
Overlapping of filter
Classification Approaches
+tare mass of
dish
+tare mass of
filter
+tare mass of
dish
44
Organic Substances
45
Organic Substances
Classification and measurement of carbonaceous organic matter
• Direct :
Based on measurement of organic carbon:
- Total Organic Carbon (TOC)
• Indirect :
Based on theoretical considerations:
- Theoretical Oxygen Demand (ThOD)
Based on measurement of oxygen consumption:
- Chemical Oxygen Demand (COD)
- Biochemical Oxygen Demand (BOD)
- Ultimate Biochemical Oxygen Demand (UBOD)
46
Direct Characterization of Organic Substances

Total Organic Carbon (TOC):
DIN EN 1085:
„Concentration of carbon present in the organic matter which is dissolved or
suspended in water“
- concentration of organic carbon in total
- The organic bound carbon is the characteristic fraction of all organic substances. The sum of
organic carbon in total (lumped parameter) is therefore a measure for the sum of all organic
substances
- In the TOC test the organic carbon is directly measured, in an instrumental test, determined through the
conversion of organic carbon into carbon dioxide (measures all carbon released in the form of CO2)
- The TOC test takes only 5 to 10 min to complete.
47
Das Messprinzip basiert auf dem Hochtemperaturaufschluss Probevolumen bzw. Gewicht und Detektorsignal, auf Basis

der Probe im Luft/O2 Strom bei 850 °C. Für thermisch stabile einer Kalibrationskurve, den Kohlenstoffgehalt.
Feststoffe können Aufschlusstemperaturen von permanent Ist in einer Probe anorganischer Kohlenstoff (Karbonat)
bis zu 1200 °C oder kurzzeitig 1800 °C erreicht werden. Dabei enthalten, kann er nach Ansäuern und Freisetzen im Spar-
wird der gesamte gebundene Kohlenstoff in CO2 überführt, ger separat als TIC bestimmt werden. Der TOC errechnet
das mit einem NDIR Detektor quantitativ bestimmt wird. Vor-
- TOC Analyzer
sich dann aus TC-TIC. Eine TOC bzw. NPOC Direktbestim-
teil dieser Methode gegenüber dem nasschemischen UV/ mung ist nach externer TIC Entfernung ebenfalls möglich.
Persulfataufschluss ist die absolute Sicherheit, dass selbst Der Sparger kann auch unmittelbar zur Direktbestimmung
stabilste Verbindungen, Partikel oder salzhaltige Lösungen flüchtiger Kohlenstoffverbindungen (POC) genutzt werden.
vollständig erfasst werden und zusätzlich die Bestimmung Dazu ist lediglich ein zusätzlicher CO2 Absorber erforderlich.
von gebundenem Stickstoff (TNb) möglich ist. Damit sind mit einem Gerät und ohne aufwändige Umbauten
Die flüssige Probe wird über eine Dosierspritze angesaugt alle relevanten Parameter der TOC-Analytik zugänglich.
und direkt in das Verbrennungsrohr injiziert, wo die kataly- Der Einsatz NO spezifischer Detektoren erweitert den
tische Oxidation erfolgt. Alternativ werden Feststoffproben Anwendungsbereich auch auf die TNb Analytik von gebun-
über einen Kugelhahn zugeführt. Nach dreistufiger Trock- denem Stickstoff. Das Gesamtkonzept ist geprägt von
nung des Verbrennungsgases und Ausgleich von Druck- kürzesten Gas- und Flüssigkeitswegen und einem Minimum
schwankungen durch einen elektronischen Durchflussregler an Ventilen mit Vorteilen für die Analysengeschwindigkeit,
erfolgt die Messung des Gesamtkohlenstoffgehaltes (TC) Nachweisstärke oder Messung partikel- und salzhaltiger
am NDIR Detektor. Ein angeschlossener PC errechnet aus Proben.
H3PO4
Luft
Probe Sparger Verbrennung Trocknung Detektion
48

Theoretical Calculation TOC by using the chemical oxidation equation
Example – TOC of glucose:
Chemical formula : C6 H12 O6
6 C + 6 O2
6 CO2
Molar weight:
C = 12 g / mol C: 12 . 6 = 72
H = 1 g / mol H: 1 . 12 = 12
O = 16 g / mol O: 16 . 6 = 96
180 g / mol
Specific TOC = 72 / 180 = 0.40 g TOC / g glucose
49

Calculation TOC by using the chemical oxidation equation
Example – TOC of glucose:
Specific TOC = 72 / 180 = 0.40 g TOC / g glucose
E.g.: 1 tea spoon of sugar (5 g glucose) dissolved

in 1 Liter of water
TOC = 5 g/L*0.4 = 2.0 g / L
50

Classification of TOC present in wastewater
TOC
Dissolved Particulate
organic carbon organic Carbon
(DOC) (POC)
• Measure for the concentration of all • Measure for the sum of all non-
dissolved (filterable) organic substances filterable organic substances
51
Indirect Characterization of Organic Substances- ThOD

Theoretical Oxygen Demand (ThOD)
• The main ecological effect of organic pollution in a water body is the decrease in the
level of dissolved oxygen
• The ThOD is the theoretical calculation of the oxygen demand if the chemical formula
of the organic matter present is known
• Organic matter of animal or vegetable origin in wastewater is generally a combination

of carbon, hydrogen, oxygen, and nitrogen (present in carbohydrates, proteins, oils and
grease)
52
Indirect Characterization of Organic Substances- ThOD

ThOD
What can be oxidized?
C CO2
H H2O
N NO2-, NO3-
S SO42-
53
Indirect Characterization of Organic Substances

Calculation ThOD by using the chemical oxidation equation
Example 1 – ThOD of glucose:
6 C + 6 O2 6 CO2
Molar mass:
C = 12 g / mol C: 12 . 6 = 72
H = 1 g / mol H: 1 . 12 = 12
O = 16 g / mol O: 16 . 6 = 96
180 g / mol
54


C6 H12 O6 + 6 O2 6 CO2 + 6 H2 O + energy
Molar mass O2 : 32 g / mol

6 . 32 g / mol = 192 g O2 / mol glucose
Molar mass glucose: 180 g glucose /mol glucose
specific ThOD = 192 / 180 = 1.067 g O2 / g glucose
55


in 1 Liter of water
specific ThOD = 192 / 180 = 1.067 g O2 / g glucose
ThOD = 5 . 1.067 = 5.3 g O2 / L
56

Calculation of the ThOD
Example 2 – ThOD of glycine:
Chemical formula Glycine (CH2(NH2)COOH )
1. Conversion of organic carbon and nitrogen to carbon dioxide and ammonia:
CH2(NH2)COOH + 1.5 O2 à NH3 + 2 CO2 + H2O
2. Oxidation of ammonia to nitrite and nitrate:
NH3 + 1.5 O2 à NO2- + H2O + H+

NO2- + 0.5 O2 à NO3-
57

Calculation of the ThOD
Example 2 – ThOD of glycerine:
Chemical formula Glycine (CH2(NH2)COOH )
ThOD = (1.5 + 2) mol O2 / mol glycine
Molar mass: 32 g/ mol O2
Specific ThOD (per mol) = 3.5 * 32 = 112 g O2 / mol glycine
Molar Mass glycine: 75.07 g glycine /mol glycine
Specific ThOD = 112 / 75.07 = 1.49 g O2 / g glycine
58

• The ThOD is a measurement for the pollution potential of a wastewater by the theoretical
calculation of the oxygen consumption that it would cause, that is, an indirect
quantification of the potential to generate an impact.
• However, in practice, wastewater has a great heterogeneity in its composition
• Trying to establish all its constituents in order to calculate the oxygen demand based on
the chemical oxidation reactions of each of the components is impossible and an
extrapolation of the data to other conditions is not possible
Measuring in the laboratory the consumption of oxygen exerted

by a standard volume of wastewater:
COD, UBOD, BOD
59
Take Home Messages
• Understand function of a municipal wastewater treatment facility
• Understand composition of municipal wastewater
• Target parameters and values for wastewater treatment
• Parameter for suspended solids
• Parameter for dissolved organic substances
60
Wastewater Characteristics II
Dr.-Ing. Benedikt Aumeier & Prof. Dr.-Ing. Jörg E. Drewes

14 December 2023

1

- Chemical Oxygen Demand (COD) -
2
Indirect Characterization of Organic Substances - COD

DIN EN 1085:
Chemical Oxygen Demand (COD) „Concentration of oxygen equivalent to the amount of dichromate consumed
when a water sample is treated with that oxidant under defined conditions“
• Equivalent for the concentration of all organic substances, independent from their composition
and biodegradability
• The COD is a lumped parameter, expressing, how much oxygen is needed for a complete
oxidation of organic substances to CO2 and H2O. Therefore, it is an indirect indication of organic
substances
• The COD is especially useful because:

• it captures the organic substances almost completely,
• can be determined at wastewater treatment plants relatively easily
• Corresponds to an electron balance for which laws of conservation hold
• directly reflects the oxygen consumption in biochemical oxidation reactions
• The COD test can be completed in about 2.5 h. To reduce the time further, a rapid COD test that
takes only about 15 minutes has been developed
3
COD - what can be oxidized?
C CO2
H H2O
N NO2-, NO3-
S SO42-
4

Theoretical calculation of COD by using the chemical oxidation equation (=ThOD)
Example 1 – COD of glucose:

C6 H12 O6 + 6 O2 6 CO2 + 6 H2 O + energy
Molar mass O2 : 32 g / mol

6 . 32 g / mol = 192 g O2 / mol glucose
Molar mass glucose: 180 g glucose /mol glucose
specific COD = 192 / 180 = 1.067 g O2 / g glucose
5

Theoretical calculation of COD by using the chemical oxidation equation (=ThoD)
Example 1 – COD of glucose:


in 1 Liter of water
Specific COD = 192 / 180 = 1.067 g O2 / g glucose
COD = 5 . 1.067 = 5.3 g O2 / L
6

COD Test
Chemical oxidation using potassium dichromate (K2Cr2O7) as oxidizing agent
- Heat for 2 hours at 148 °C
- Titrate non-consumed potassium dichromate with ferrous ammonium

sulfate or photometric detection of non used potassium dichromate
- Calculate the oxygen consumed during digestion
7

Classification of COD present in wastewater I Total COD
Biodegradable Non-biodegradable
COD (bCOD) COD (nbCOD)
Readily Slowly Non- Non-

biodegradable biodegradable biodegradable biodegradable
(soluble) (particulate) soluble particulate
(rbCOD) (sbCOD) COD COD
(nbsCOD) (nbpCOD)
• can be quickly • particulate and colloidal
• Organic material,
assimilated by • must first be dissolved by • For domestic
biomass nbpCOD
extracellular enzymes wastewater the WWTP
corresponds to
• “true” dissolved • are thus assimilated at influent nbsCOD ≈
sCODe (sCODe = non-biodegradable
much slower rates
volatile suspended
activated-sludge
process effluent) when solids (nbVSS)
operated at an SRT >
4d
8

Classification of COD present in wastewater II Total COD
Particulate (pCOD) Soluble (sCOD)
•Colloidal and sCOD = rbCOD +

suspended solids part of colloidal
COD + nbsCOD
non non-
biodegradable biodegradable
biodegradable biodegradable
particulate soluble
particulate soluble
(bpCOD) (bsCOD)
(nbpCOD) (nbsCOD)
• slowly
biodegradable • rbCOD + part of colloidal COD
• rbCOD is “true” soluble COD
• Biodegradable colloidal COD is
slowly biodegradable
9

Particle Size Ranges
algae
bacteria
Bacterial flocs
cell fragments
viruses
protein
DNA Visible to the naked eye
fatty acids
ammonium, *Filter pore size
nitrate, etc.. 0.45 µm* if 0.45 µm is
used to
settleable distinguish
dissolved colloidal between
suspended particulate and
dissolved
10-6 10-5 10-4 10-3 10-2 10-1 100 10+1 10+2 µm fraction
1 µm=0.001 mm 10

Importance of readily and slowly biodegradable fractions
(illustated by an example of a 3 staged cascade):
tank 1 tank 2 tank 3 Secondary clarifier
rate mg/L h
Relative oxygen
rbCO • in case tank 1 is aerated: High rbCOD in

D the influent causes higher oxygen demand
(more aeration needed)
uptake
• in case tank 1 is anoxic (preanox

denitrification): High rbCOD causes higher
uptake rate mg/L h
Relative oxygen
sbCOD denitrification rates
11

Importance of readily and slowly biodegradable fractions
• Carbonaceous constituents measured by BOD or COD analyses are critical to the activated-sludge
process design. Higher concentrations of degradable COD may e.g. result in
1. a larger aeration basin volume,
2. more oxygen transfer needs, and
3. greater sludge production
• Especially the rbCOD fraction of the COD has a direct effect on the biological process performance
• The biodegradable fractions of the COD are important for biological treatment and must be evaluated
Biochemical oxygen demand (BOD)
12

- Biomass -
13
Indirect Characterization of Organic Substances - Biomass

Biomass
• The biological treatment of municipal wastewater is achieved by using mixed cultures of microorganisms
composed of different types of microorganisms, the exact composition of the different types being
unknown
• The term „biomass“ therefore describes a very complex mix of different types of microorganisms, having
(very) different characteristics regarding their activity
• The properties of this complex mixed biomass depends on the conditions characterized by the process
type and the characteristics of the wastewater
• Biomass is mostly organic material and can be represented by
• volatile suspended solids (VSS)
• particulate COD (pCOD)
Other parameters to represent biomass include DNA, ATP, protein content, ….

14

Importance of biomass for biological treatment process design
• Requires an understanding of the biochemical activities of microorganisms
• Microorganisms
- consume substrate,
- carry out oxidation-reduction reactions,
- are subject to growth (by the production of additional cells), and decay processes
• Therefore, biomass is produced continuously (by consumption of the substrate in the wastewater) and
decomposted (biodegraded)
15

Characterization of biomass
BIOMASS
(part of pCOD and VSS)
active inactive
• e.g. • resulting from decay

- Heterotrophic bacteria, processes of biomass
- Autotrophic bacteria (cell debris)
- ….
16

Fundamental transformation processes:

5 CH3OH + 6 NO3- è 5 CO2 + 7 H2O + 3 N2 + 6 OH-
17

- BOD -
18
Indirect Characterization of Organic Substances – BOD

• Biochemical Oxygen Demand (BOD)
DIN EN 1085:
„Concentration of dissolved oxygen consumed under specified conditions (t days at 20°C
with or without nitrification inhibition) by the biological oxidation of organic and/or
inorganic matter in water“
• Equivalent for the concentration of the biological degradable fraction of organic substances
• The BOD is a lumped parameter, expressing, how much oxygen is needed to stabilize, through
biochemical processes, the carbonaceous matter. Therefore, it is an indirect indication of the
biodegradable organic carbon present
19

Biochemical Oxygen Demand after 5 days (BOD5)
• If sufficient oxygen is available, the aerobic biological decomposition of an organic waste will continue until
(nearly) all of the waste is consumed
• (Nearly) complete stabilization takes, in practical terms, various days (around 20 days of more for domestic
sewage)
• However, to shorten the time for the laboratory test, and to allow a comparison of the various results,
some standardizations were established:
- The determination is undertaken on the 5th day
- The BOD test is carried out at a temperature of 20°C
• The most widely used parameter of organic pollution applied to both wastewater and surface water is
the 5-day BOD (BOD5 or BOD520)
• The 5 day duration has mainly historical background
20



5 CH3OH + 6 NO3- è 5 CO2 + 7 H2O + 3 N2 + 6 OH-
21

BOD test; dilution method
• prepare dilution water (20 °C, oxygen saturated)
• measure dissolved oxygen concentration at time zero (= D1)
• incubate for 5 days (dark, 20 °C)

• measure dissolved oxygen concentration after 5 days (= D2)
• calculate BOD5 = (D1 – D2) / volume
22

Example for a BOD test for a domestic wastewater:
Oxygen
[mg/l]
time [d]
23

Example for a BOD test for a domestic wastewater
Oxygen demand
[mg/l]
BOD5
5d time [d]
24
Typical BOD 5 Water

Concentrations [mg/L]
3-30 River water
3-60 Biologically treated ww
40-600 Low polluted industrial ww
100-300 Municipal untreated
wastewater (ww)
400-1,200 Polluted industrial ww
1,000-6,000 Highly polluted industrial
wastewater
25

Analysis of the BOD- test
Oxygen [mg/l]
Oxygen
consumed until
time t (BOD
measured)
time [d]
- During the test three more or less distinct activities occur:

(1) a portion of the waste is oxidized to end products to obtain energy for cell maintenance and the synthesis of new cell tissue
(2) Simultaneously, some of the waste is converted into new cell tissue using part of the energy released during oxidation
(3) decay of cells
26

What happens after 5 days?
- At the end of the fifth day the stabilization of the organic material is still not complete. If the test was
continued, degradation will go on (however, continuing at slower rates).
- Biochemical oxidation theoretically takes an infinite time to go to completion (the rate of oxidation is assumed
to be proportional to the amount of organic matter remaining).
Ultimate biochemical oxygen demand (UBOD or BODu)
27

- Ultimate BOD -
28
Indirect Characterization of Organic Substances - UBOD

Ultimate biochemical oxygen demand
(BODu)
Oxygen demand [mg/l]

BODu
• Depending on the type of wastewater in

the 5 day period, 60 to 70% of oxidation BOD5
95 – 99%
is complete and oxidation will still go on.
Within a 20 day period, the oxidation of
the carbonaceous organic matter is about
95 to 99 % complete.
5d time [d] 20 d
• For domestic sewage, it is considered that after 20 days of the test the stabilization is practically
complete. Therefore, in practice the BODU can be determined at 20 days, or estimated by following the
BOD course as a function of time and calculating the UBOD.
• The concept of the test is similar to the standard BOD of 5 days, varying only with the final period of
determination of the dissolved oxygen concentration (e.g. 20 days).
29
Analysis of the BOD - test
substrat
e
biomass
Cell debris
time
30

Analysis of the BOD - test
31

Modelling biochemical oxygen demand (highly simplified!)
• Degradation is assumed to be proportional to BOD remaining:

dBODr
= -k1 × BODr
dt
Analytical solution after integration between t=0 and t=t:
BODr = UBOD × (e - k1t )

BODt = UBOD - BODr
BODt = UBOD × (1 - (e - k1t ))
BODr = amount of waste remaining at time t (expressed in oxygen equivalents, [mg/l]
k1 = first-order reaction rate constant (usually base e), [1/d]; typical (at 20°C): 0.23 1/d; untreated WW
(at 20°C): 0.12 to 0.46 1/d; effluents (at 20°C): 0.12 to 0.23 1/d
UBOD = total or ultimate carbonaceous BOD, [mg/l]
t = time [d]
32
Modelling biochemical oxygen demand (highly simplified !)
Temperature adjustment:
- if the temperature is not 20°C, k can be adjusted:
k1,T = k1, 20 ×q (T -20)

θ = 1.056 between 20 and 30°C, and 1.135 between 4 and 20”C
33

Example for a fitted BOD curve for the determination of k, UBOD and bCOD
Source: Roeleveld and van Loosdrecht (2002) 34


Average results of the performed wastewater characterization for 21 different WWTP in the
Netherlands
kmin = 0.33
kmax = 0.7


Example for the relation between the measured k value and the slowly biodegradable COD fraction
SS = readily biodegradable COD

XS = slowly biodegradable COD
SS+XS = total biodegradable COD

Example 2: Determine the 1-day BOD and ultimate BOD for a wastewater
whose 5-day BOD (20°C) is 200 mg/L. The reaction constant k
(base e) = 0.23 1/d.
What would have been the 5-day BOD if the test had been
conducted at 25°C ?
37

Solution to Example 2 (Calculating UBOD):
given: k1, BOD5, T
BODt = UBOD - BODr BODr = UBOD × (e - k1t )
At t = 5d BOD = 200 mg/l
BOD5 = UBOD -UBOD × (e - k1t )

- k1t
BOD5 = UBOD(1 - (e ))
k1, 20 = 0.23
200 = UBOD(1 -(e -0.23×5 ))

à UBOD = 293mg / l
38

Illustration of Example 2:
BODt = UBOD × (1 - (e - k1t ))

BODr
UBO
D
BOD5
39

Solution to Example 2 (Calculating BOD1):
given: k1, BOD5, T, t=1d
calculated: UBOD
BODr = UBOD × (e - k1t ) BODt = UBOD × (1 - (e - k1t ))
At t = 1d :
- k1t
BODt = UBOD(1 - (e ))
BOD1 = 293(1 - 0.795)

à BOD1 = 60mg / l
40

Illustration of Example 2:
BODt = UBOD × (1 - (e - k1t ))
BOD1
41

Solution to Example 2 (Calculating BOD5 at 25°C):
given: k1 at 20°C, T
calculated: UBOD
BOD5 at T= 25°C :
(T -20 )
k1,T = k1, 20 ×q
θ = 1.056
k1, 25 = 0.23 ×1.056(25-20)
à k1, 25 = 0.302
BOD5 = UBOD(1 - (e - k1t ))
à BOD5 = 228mg / l
42

Illustration of the influence of k1
k1=0.46
BODt
k1=0.23
k1=0.12
43

Illustration of Example 2 (temperature change):
T=25°C BODt = UBOD × (1 - (e - k1t ))

T=20°C
Time [d]
44

Illustration of Example 2 (extreme temperature change):
T=30°C, k1,20 =0,23 BODt = UBOD × (1 - (e - k1t ))
T=4°C, k1,20 =0,23
Time [d]
45
Indirect Characterization of Organic Matter

Nitrification in the BOD test
46

Nitrification during the BOD-test
• Non-carbonaceous matter, such as ammonia, is produced during the hydrolysis of proteins and
nitrification may occur during the BOD test
• The oxygen demand associated with the oxidation of ammonia to nitrate is called the nitrogenous
biochemical oxygen demand (NBOD)
• When nitrification occurs in the BOD test, erroneous interpretations of treatment operating data
are possible
• When nitrification occurs in the BOD test, the measured BOD value will be higher than the true
value due to the oxidation of carbonaceous matter
47



5 CH3OH + 6 NO3- è 5 CO2 + 7 H2O + 3 N2 + 6 OH-
48

Influence of nitrification on oxygen demand in a BOD test for a municipal wastewater
Oxygen demand [mg/l]

The reproductive rate of the
nitrifying bacteria is slow. It
NBOD
normally takes from 6 to 10
days for them to reach
significant numbers to exert
a measureable oxygen
Carbonaceous BOD
demand
A sufficient number of nitrifying
bacteria is present initially
5 days time [d]
BOD due to oxidation of carbonaceous material. When it is referred to the
oxygen demand by the oxidized carbon in the test (when nitrification is
suppressed) the oxygen demand is called carbonaceous BOD (CBOD)
49

Overcoming the effects of nitrification in a BOD test
- by using various chemicals to suppress the nitrification reactions, or
- by treating the sample to eliminate the nitrifying organisms.
original sample
BOD [mg/L]
Sample with inhibitor for nitrification
1 2 3 4 5
time [d]
Source: BSB-Fibel, Fa. WTW 50

Indirect Characterization of
Organic Matter Interrelationships
51
Indirect Characterization of Organic Substances - Interrelationships

Interrelationship between representatives of organic matter in a WWTP
influent
non biodegradable remaining biomass,
and organic matter remaining cell
COD
after 5d debris
(nbCOD)
(additional
nbCOD)
COD remaining
cell debris
bCOD (additional UBOD
BOD5 nbCOD)
after 5d
• The ratios vary as the wastewater passes along the various units of the treatment plant (the tendency is for
the ratio COD/xxx to increase); xxx= bCOD, BOD5 or UBOD
• The higher the treatment efficiency, the higher the value of the ratio of COD to the other values becomes
52
Relationship between UBOD and COD
• The value of the ultimate carbonaceous UBOD value is usually not the same as the COD, because
1. Some organic substances are difficult to oxidize biologically, but can be oxidized chemically
2. Some inorganic substances can be oxidized by dichromate (in the COD-test this increases the
apparent organic content of the sample
3. certain organic substances may be toxic to the microorganisms used in the BOD test
4. Some of the biomass can still remain as cell debris (and active cells) in the UBOD test (even after 20
days)
53

UBOD and BOD5
Origin UBOD/BOD5
High concentration sewage 1.1 – 1.5 a value of 1.6 is

Low concentration sewage 1.2 – 1.6 commonly used
Primary effluent 1.2 – 1.6
Secondary effluent 1.5 – 3.0
Source: von Sperling (2005)
Origin UBOD/BOD5
Untreated 1.1 – 2.2

(k1 = 0.12 – 0.46)
Effluent from biological treatment 1.46 – 2.2
(k1 = 0.12 – 0.23)
Calculated via UBOD/BOD = 1/(1-e^(-k1*5))
54

Relationship between COD and BOD5
Oxygen – used by microorganisms for energy
BOD5 generation
CODtotal
CODresidual
residual-COD
- biomass (growth of microorganisms that use part of
Municipal wastewater: the organic substances to build up their biomass
Typical proportion:
- refractory organic substances (non biodegradable
COD / BOD5 : ≈ 1.7 - 2 matter)
(However, for industrial wastewater,
the ratio can vary widely).
COD / TOC : ≈ 2.5
55
COD and BOD
Origin BOD/COD
After primary settling 0.4 – 0.6
Final effluent 0.1 – 0.3*
*CBOD/COD
Source: adapted from Metcalf & Eddy (2003) 56


Evaluation of wastewater by the ratio of COD and BOD5
BOD5 / COD ratio Evaluation
BOD5 / COD > approx. 0.4 to 0.5 biodegradable fraction is high,

good indication for biological treatment
approx 0.3 < BOD5 / COD poor biological degradability, waste may
have some toxic components or
< approx 0.5 acclimated microorganisms may be
required in its stabilization, treatability
studies are recommended to verify
feasibility of biological treatment
BOD5 / COD < approx 0.25 non-biodegradable fraction is high,
possible indication for physical-chemical
treatment
57
TOC and BOD5
Origin BOD/TOC
After primary settling 0.8 – 1.2
Final effluent 0.2 – 0.5*
*CBOD/COD Source: Metcalf & Eddy (2003)
58
Example 3: Determine the theoretical TOC/COD ratio for the following compound C5H7NO2.
59
Solution to Example 3:
C5H7NO2 + 5 O2 à 5 CO2 + NH3 + 2H2O
C5H7NO2 : mw = 113
O2 : mw = 32
COD = 5 * 32/113 = 1.42 mg O2 / mg C5H7NO2
TOC = (5 * 12)/113 = 0.53 mg TOC / mg C5H7NO2
TOC 0.53 COD

à = = 0.37 = 2.7
COD 1.42 TOC
Note: C5H7NO2 is often assumed to be a typical composition of biomass.
60

Possible properties of COD Bio- Non-bio- dissolved Suspended
fractions degradable degradable (soluble) (particulate
)
organic
inorganic
dissolved
(soluble)
Suspended
(particulate
)
61

Properties of some COD Bio- Non-bio- dissolved Suspended
fractions degradable degradable (soluble) (particulate
)
bCOD = biodegradable COD
bCO nbCOD rbCOD pCO
organic
D
rbCOD nbsCOD D
nbpCOD rbCOD=readily biodegradable COD
nbsCOD
bpCO nbpCOD bpCO
D D nbpCOD = nonbiodegra-dable
inorganic particulate COD
bpCOD=biodegradable particulate COD
dissolved
rbCOD nbsCOD nbsCOD=nonbiode-gradable soluble
(soluble)
COD
pCOD=particulate COD
Suspended nbpCOD nbCOD=nonbiodegradable COD
(particulate) bpCO
D
Note: soluble COD (sCOD) = rbCOD + colloidal COD + nbsCOD); due to the method used (filter with 0.45 μm pore size); but
only the rbCOD is quickly assimilated by the biomass
62
Nitrogenous Compounds
63
Plant nutrients
- ammonium NH4 +
-
- nitrite NO2
-
- nitrate NO3
- phosphate PO43-
64
Nitrogenous Compounds in the Environment

• The elements nitrogen and phosphorous are essential nutrients for algae leading, under
certain conditions, to eutrophication of lakes and reservoirs
• Nitrogen can lead to dissolved oxygen consumption in the receiving water body due to
the processes of nitrification
• Nitrogen in the form of free ammonia is directly toxic to fish. At high pH-values, a high
amount of ammonia is produced
• Nitrite is an intermediate product in the nitrification process and usually low in concentration,
however highly toxic to fish
• Nitrate is associated with illnesses, therefore undesirable in water and is only in limited
amounts tolerable in drinking water
65
Nitrogenous Compounds in a Treatment Plant
• As nitrogen and phosphorous are essential nutrients for the growth of microorganisms,
they are needed for the treatment of wastewater by biological processes
• Nitrogen in the process of nitrification leads to oxygen consumption
• Nitrogen in the process of denitrification can lead to the deterioration in the settleability
of the sludge in the secondary clarifier
66
Characterization of nitrogen found in wastewater
• The most common and important forms of nitrogen in wastewater are Ammonia (NH3), Ammonium (NH4+),
Nitrogen gas (N2), Nitrite ion (NO2-) and Nitrate ion (NO3-)
• Urea, also present in fresh wastewater, is converted rapidly to ammonium carbonate, and is seldom found in
untreated municipal wastewaters
• Nitrite is an intermediate product of microbial processes and is usually not present in high concentrations.
In the presence of O2 nitrogen usually accumulates to nitrate
• N2 has poor solubility in water
• The age of wastewater is indicated by the relative amount of ammonia that is present
• The determination of the prevailing form of N in a water body can provide indications about the stage of
pollution caused by an upstream discharge of sewage
67
Relative distribution of the forms of nitrogen under different conditions
Condition Prevailing form of N
Raw wastewater • Organic N
• Ammonia
Recent pollution in a water course • Organic N
• Ammonia
Intermediate stage in the pollution of a water course • Organic N
• Ammonia
• Nitrite (low)
• Nitrate
Remote pollution in a water course • Nitrate
Effluent from a treatment process without nitrification • Ammonia
Effluent from a treatment process with nitrification • Nitrate
Effluent from a treatment process with nitrification and • Low concentrations of all
denitrification forms of nitrogen
Source: von Sperling (2005)
68
Nitrogenous Compounds N
Characterization of nitrogen
present in wastewater TKN nitrite nitrate
• usually
low
Organic N ammonium,
ammonia
non - bio-
biodegradable degradable
Soluble Particulate Soluble Particulate
• usually • Associated with the •Quickly • slowly

negligible non-biodegradable biodegradable biodegradable
carbonaceous matter,
involved in the
biomass
69
Nitrogen distribution in raw (untreated) domestic sewage
NOx NOx NOx
Total
Ammonia TKN soluble inorganic
60 to 70% of nitrogen
TKN the influent
N total (TIN)
TKN
N org N
org
TKN
particulate
(adapted from von Sperling, 2005 and IAWQ, 1995) 70

Nitrogenous Compounds - Ammonia

• The inorganic nitrogenous matter is represented by ammonia, both in the form of NH3 and as NH4+
• Ammonia in the influent wastewater is mainly derived from urea, which is rapidly hydrolyzed and rarely found
in raw sewage
• Hydrolysis and ammonification reactions begin in the collection and interception sewerage system
• In the usual range of pH, near neutrality, the ammonia present is practically in the form of NH4+
Source: Metcalf & Eddy (2003) 71

Phosphorous
72
Phosphorous
• Total phosphorus in domestic sewage is present in the form of phosphates
• Phosphorus is an essential nutrient for the growth of the microorganisms responsible for
the stabilization of organic matter
• Phosphorus is an essential nutrient for the growth of algae, eventually leading, under
certain conditions, to the eutrophication of lakes and reservoirs
73
Phosphorous
Characterization of phosphorous P total
species present in wastewater
Organic P Inorganic P
Bound to organic Main source from detergents
compounds, and other household
physiological origin products
Particulate P Soluble P Soluble P
orthophosphates
polyphosphates Available for biological
Subject to hydrolysis metabolism without
further breakdown
74
Phosphorous
Phosphorus in raw (untreated) domestic sewage
P inorg (poly +
P orthophosphate)
soluble
P
total
P P
particulate org
(adapted from IAWQ, 1995) 75

Example 4: Given the wastewater characteristics in the table, determine the concentrations for
- bCOD (biodegradable COD)
- nbpCOD (nonbiodegradable particulate COD)
- sbCOD (slowly biodegradable COD)
- FSS
- the fraction of organic nitrogen in VSS
Constituent (influent) Concentration mg/l
BOD 195
COD 465
rbCOD 80
TSS 220
VSS 200
TKN 40
NH4-N 26
Constituent (effluent)
sCODe 30
sON (soluble org N) 1.2
76
Determine the concentrations for
bCOD, nbpCOD,sbCOD, FSS, the fraction of organic nitrogen in VSS
table.: BOD, COD, rbCOD, TSS, VSS, TKN, NH4-N, sCODe, sON
bCOD = UBOD + 1.42 × f d × (YH ) × bCOD

bCOD / BOD = UBOD / BOD + 1.42 × f d × (YH ) × bCOD / BOD
bCOD/BOD = 1.6 (under the assumption that UBOD/BOD=1.46)
à bCOD/BOD =1.6 is often assumed à bCOD = 1.6 * BOD = 1.6*195 = 312 mg/l
nbpCOD = nbCOD – nbsCOD
For domestic wastewater nbsCOD ≈ sCODe (sCODe = activated-sludge
process effluent) when operated at an SRT > 4d
à nbpCOD = nbCOD – sCODe
nbCOD = COD – bCOD = 465 – 312 = 153 mg/l
à nbpCOD = nbCOD – sCODe = 153 - 30 = 123 mg/l
sbCOD = bCOD – rbCOD = 312 -80 = 232 mg/l
FSS = TSS – VSS = 220 - 200 = 20 mg/l
77
Determine the concentrations for
bCOD, nbpCOD, sbCOD, FSS, the fraction of organic nitrogen in VSS
table.: BOD, COD, rbCOD, TSS, VSS, TKN, NH4-N, sCODe, sON
TKN = orgN + NH4-N

orgN = sON + pON ( = particulate organic N)
TKN = sON + pON + NH4-N
àpON = TKN - sON – NH4-N = 40 – 26 -1.2 = 12.8 mg/l
àfraction of organic N in VSS = pON / VSS = 0.064 gN/gVSS
78
Temperature
79
Temperature and Temperature Effects

• Temperature is a crucial factor in biological processes
• Temperature effects include effects on chemical reactions, equilibrium constants,

solubility constants, biological reaction-rate constants, etc. (all of them are dependent on
temperature)
• Different (mean) temperature can tremendously influence the performance and the design of
wastewater treatment plants:
- tank volumes
- sludge production
- oxygen demand
80
Temperature
Temperature Effects, e.g. tank volume in the design of a WWTP
(15 design models from eight countries)
Tank volume [m³]
Temperature [°C] Wichern et al. (2004)

With the data set used:
• Differences of more than 200% between models at 10°C were observed
• 10°C Temperature increase could result in a decrease of about 50%
• it is recommended to use design procedures for other countries / conditions only if they have
been tested for these conditions
81
Temperature
Wastewater temperature variations
• Temperature of wastewater is commonly higher than that of the local water supply, because of the
addition of warm water from household and industrial activities.
• The temperature variation of the water over a year is delayed compared to the time of the year and the
temperature of the air. Seasonal heating and cooling is delayed.
• Observed wastewater temperatures are higher than the local air temperatures during most of the year
and are lower only during the hottest summer months.
• Variations of wastewater temperature over one day are caused by the change of the portion of warm
water in the wastewater.
• Knowing the temperature extreme values is very important for wastewater treatment plant design. If
the plant is big, often daily mean values are sufficient (due to the temperature equalization of the
variations over one day in big tank volumes).
82
Wastewater Temperature Variations

Daily mean temperature of raw wastewater in a WWTP in Switzerland (in °C) over one year:
Seasonal variations are lower and delayed compared to air temperature and season
83
Temperature
Wastewater temperature ranges based on an international study
Country Wastewater Temperature BOD BOD N

Influent [°C] [mg/l] [g/ capita d] [mg/l]
[l/(capita d)]
Brasil 120 - 180 10 - 28 108 - 299 29 27 - 64
China - 5 - 25 44 - 386 - 35
Indonesia - - - 27 - 70 -
Morocco - - - 32 - 40 9
South Africa 10 - 800 10 - 30 250 - 400 - 35 - 80
Thailand - 26 - 33 20 - 400 - 17 - 20
USA - 1 - 25 160 - 360 - -
http://www.ptka.kit.edu/downloads/ptka-wte-w/WTE-W-Berichte-2010_Leitfaden_Abwassertechnologie_Laendern.pdf
84
Fundamentals of Microbial Metabolism, Stoichiometry,

Process Kinetics, and Biological Process Models

11 January 2024
1
Fundamentals of Microbial Metabolism,

Stoichiometry, Process Kinetics and Biological
Process Models
- Microbial Metabolism -
2
Microbial Metabolism
The Role of Microorganisms
• Biological processes in wastewater treatment consist of mixed communities with a wide variety of
microorganisms, including
• bacteria,
• protozoa,
• fungi,
• rotifiers,
• and possibly algae
• Bacteria constitute the largest

and most important group in
biological wastewater treatment
systems
3
EP
S
adapted from von Sperling (2005)
Simplified schematic representation of a bacterial cell

Genetic material • DNA and plasmides
Ribosome • location of enzyme synthesis
Cell membrane • external boundary,
• has an important role in selecting the substances that can leave or enter the cell,
• location where energy is gained
Cell wall • another external layer, keeps the form of the cell,
• protection against mechanical impacts and osmotic alterations
Extracellular matrix • composed of polysaccharides and polypeptides that is often found around the cell
(EPS) • plays an important role in e.g. the formation of bacterial flocs that are important in
wastewater treatment
4
Sources needed for microbial growth
Reproduction and proper functioning of an organism require sources of
• energy
• carbon
• nutrients
- inorganic elements
- organic nutrients (growth factors)
Carbon source New cells
Electron donor other products
5
Energy sources for microorganisms
• Microorganisms require chemically available energy, to
• carry out synthesis,
• enable mobility, a big portion of this energy is supplied by
• maintain inner structure and osmotic barriers • redox reactions, or
• light
Carbon sources for microorganisms

• obtained from
• organic matter, or
• carbon dioxide
Nutrients
• Nutrients may at times be the limiting factor for microbial cell synthesis and growth
• Principal inorganic nutrients: N, S, P, K, Mg, Ca, Fe, Na, Cl
• Minor nutrients
6
Metabolism of microorganisms and enzymes
• Metabolism: the chemical processes that simultaneously take place in the cell
• The chemical transformations occur in a sequence of diverse and complex intermediate reactions, each catalyzed by
a specific type of enzyme
• Enzyme types:
- Intracellular enzymes
Most of the enzymes, located inside the cell, aid in the consumption of soluble matter
- Extracellular enzymes
Some enzymes are released into the external medium,
lead to hydrolysis that converts large and complex substrate molecules in order to make them
available for passing through the cell membrane
7
Carbon source New cells
Electron donor other products
aerobic, aerobic, anaerobic,

heterotrophic autotrophic heterotrophic
8
Simplified Scheme of Microbial Metabolism

The reaction sequences within the cell can be divided into the following four interrelated steps:
Decomposition, synthesis, energy production, and storage
Storage
Nutrients,
carbon sources Synthesis
Decom
pos. New cells
Electron Energy
donor e-
Electron acceptor
Products
Metabolism works only if substrate AND electron acceptors are available ! 9

Decomposition
Serves for the retrieval of material for the synthesis
of cell components and electrons that are needed
for gaining energy
e-
Energy production,
Stepwise transfer of electrons to the electron
acceptor to gain energy
Synthesis
Supplied with the energy
Production of the components of the new cells (reproduction)
Production of substances, that are needed to build up the storage
Production of substances, that serve to repair cell components
Storage: Serves for building up reserves for a survival in periods of shortage
10
Classification of Microorganisms
Sources Microorganism type

Energy source
a) Radiation (light) Phototrophic
b) Chemical reactions Chemotrophic
Electron donor (substrate being oxidized)
a) Inorganic (e.g. NH4+, NO2-,H2,…) Lithotrophic
b) Organic compounds Organotrophic
Carbon source for biomass synthesis
a) Carbon dioxide, CO2 as main source Autotrophic
b) Organic compounds Heterotrophic
11
Conditions in the environment
Energy production Prevalent Conditions in
the environment
Available electron acceptor being reduced
Respiration
a) O2 Aerobic
b) Nitrate, NO3-, nitrite, NO2-, no oxygen Anoxic
c) Sulfate, SO42-, CO2, no oxygen, no nitrate Anaerobic
Fermentation
d) Organic compounds, no oxygen, no nitrate Anaerobic
• In biological wastewater treatment almost the whole spectrum of microorganisms is engaged in transformations
12
Example
• A microorganism, oxidizing ammonium to nitrite that needs (obligatory) oxygen as electron
acceptor to synthesize its biomass from carbon dioxide is called:
Obligate aerobic, chemo-litho-autotrophic
13
Examples
chemo-organo-heterotrophic aerobic bacteria
(coh-aerobic)
responsible for the elimination of organic wastewater constituents
chemo-organo-heterotrophic anoxic bacteria

(denitrifiers)
responsible for the transformation of nitrate into nitrogen gas (N2)
chemo-litho-autotrophic aerobic bacteria

(nitrifiers)
responsible for the transformation of ammonium into nitrate (or nitrite)
14
Preferences of organic matter decomposition
• The objective of the energetic metabolism is to conserve as much energy as possible in a form available to a cell
• When various electron acceptors are available in the medium, the microorganisms use the one that produces the
highest quantity of energy:
(1) Dissolved oxygen is used first under aerobic conditions, after it is exhausted
(2) When dissolved oxygen is not available and nitrate is available, the organism that are capable of using nitrate start to
take over (anoxic conditions)
(3) When there is no nitrate, strict anaerobic conditions occur, where sulfates are used and reduced to sulfides, and
CO2 is converted into CH4 (methane) (anaerobic conditions)
• Since more energy is released through aerobic reactions than through anaerobic reactions, the aerobic organisms
reproduce faster than the anaerobic organisms
15
Main routes of organic matter decomposition in the presence of different electron acceptors:
CO2 H2O
aerobic conditions
O2
CxHyOz
Energy gain
Organic matter
anoxic conditions NO3 -
CO2 N2
SO4 2-
CO2
anaerobic conditions
CO2 H2S
CO2 CH4
Source: von Sperling (2005) 16

Microbial Metabolism/Biochemical Pathways
17
Detection of SARS-CoV-2 by Biomarker Project

- In silico tests - - Primerauswahl -
A B C D E F G H I J K L M N O P Q Targetgen Sequenz Referenzen
FSSRFSSRFSSRFSRFSRFSRFSSRFSRFSRFSSRFSRFSRFSRFSRFSRFSRFSR
Corona_229E ORF6-8 GACCCCAAAATCAGCGAAAT CDC 2020; Nemudryi et al. 2020; Wu
SARS-CoV-2 Genome
Corona_avian M Nukleocapsid FAM-ACCCCGCATTACGTTTGGTGGACC- et al. 2020; Medema et al. 2020;
Corona_bat E QYS Rimoldi et al. 2020; Peccia et al.
Corona_HKU1
Corona_mers ORF1a ORF1ab S
ORF3 N
ORF10
N TCTGGTTACTGCCAGTTGAATCTG 2020; Randazzo et al. 2020a;
Corona_murine
Randazzo et al. 2020b
5`UTR Corona_NL63
3`UTR ACAGGTACGTTAATAGTTAATAGCGT Corman et al. 2020; Wurtzer et al.
Corona_pigeon
Hüllprotein
0 10000
Corona_porcine
enterovirus
20000 30000 FAM-ACACTAGCCATCCTTACTGCGCTTCG- 2020; Bar Or et al. 2020
norovirus E QYS
SARS 2020 ATATTGCAGCAGTACGCACACA
SARS before 2016
SARS Africa
Replikase-Polyprotein TGGGGYTTTACRGGTAACCT Chu et al. 2020; Etievant et al. 2020;
SARS Brazil (Open Reading Frame) VIC-TAGTTGTGATGCWATCATGACTAG- Curti et al. 2020; Sah et al. 2020
SARS Canda QYS
SARS China
SARS France
ORF AACRCGCTTAACAAAGCACTC
SARS Germany RNA-abhängige RNA- GTGARATGGTCATGTGTGGCGG Corman et al. 2020; Wurtzer et al.
SARS India
SARS Italy
Polymerase CAGGTGGAACCTCATCAGGAGATGC 2020; Alpaslan Kocamemi et al.
SARS Peru CARATGTTAAASACACTATTAGCATA 2020b; Alpaslan Kocamemi et al.
SARS Russia
SARS Spain
RdRP 2020a; Westhaus et al., 2020
SARS USA
SARS Reference
No Missmatch
1 Missmatch
→ The detection is based on 3-4 gene fragments
2 Missmatches
3-4 Missmatches
5+ Missmatches
Ho et al. (2022), Water Research 210, 117977

Biomarker Results Berchtesgaden (21.906 PE)

– rolling average
12/1/20 1/1/21 2/1/21 3/1/21 4/1/21 5/1/21
120 140
SARS-CoV-2
Neuinfektionen in 7 Tagen
Biomarker: E-Gen 120
100
SARS-CoV-2 Biomarker Konzentration

pro mL Abwasser
Biomarker: Durchschnitt aus 3 Zielgenen
Genkopien pro mL Abwasser

100
Neuinfektionen in 7 Tagen
pro mL Abwasser
80 Gleitender Mittelwert (Durchschnitt)
80
60
60
40
40
20
20
0 0
12/2020 1/2021 2/2021 3/2021 4/2021 5/2021
• Sampling 2x/week provides reasonable representation of infection pattern

• Biomarker abundance approx. 7 days ahead of official case data
Rossmann et al. (2021), Detusches Ärztebaltt 19

Biomarker Results in Karlsruhe

01.06.2020 Infection 18.12.2020
number ddPCR (E, ORF)
01.06.2020 Infektionszahle
18.12.2020
500
0 10
0
50
40
30
20
0…
0…
0…
0…
1…
1…
1…
0…
0…
0…
0…
0…
0…
0…
n
o…
350 50
cti
RS
-…
fe
In
A
S
Aktive Fälle pro 100.000 Einwohner
RNA proconcentration
Infections per 100,000 inhabitants
mL Abwasser
300 A
Infection data not shifted 40
250 Not corrected
(original data)
per mL sewage
200 30
Biomarker
150 20
SARS-CoV-2
100
SARS-CoV-2
10
50
0 0
06/20 07/20 08/20 09/20 10/20 11/20 12/20 01/21 02/21 03/21 04/21 05/21 06/21 07/21
01.06.2020 18.12.2020
350 50
mLconcentration
Infections per 100,000 inhabitants
B
Aktive Fälle pro 100.000 Einwohner
300
Abwasser
InfectionCase
data shifted by 40
250 16 days
numbers
per m L sewage
200 shifted by 16 30
arker
days
RNA pro
150
SARS-CoV-2 Biom
20
100
SARS-CoV-2
10
50
0 0
06/20 07/20 08/20 09/20 10/20 11/20 12/20 01/21 02/21 03/21 04/21 05/21 06/21 07/21
Ho et al. (2022), Water Research 210, 117977

20
Detection of Variants of Concern (VoCs) - Karlsruhe
Anteil Alpha, Beta, Gamma, Mu, (Omikron) - S:N501Y (S:N501Y / Wildtyp) Anteil Alpha
50Diagrammtitel
ddPCR 50
Anteil
0 Delta und OmikronDiagrammtitel
- S:T478K (S:T478K / Wildtyp) RKI 0 Anteil Delta
Anteil Omikron (Omikron /1E, ORF) Anteil
1 Omikron
120%
100%
80%
Anteil Variante
60% Alpha Delta Omikron

40%
20%
0%
11/20 12/20 01/21 02/21 03/21 04/21 05/21 06/21 07/21 08/21 09/21 10/21 11/21 12/21 01/22 02/22 03/22 04/22
Ho et al. (2022), Water Research 210, 117977 21

Detection of Variants of Concern in Bavaria
Source: https://www.bay-voc.lmu.de/abwassermonitoring 22
Environmental factors
• Environmental conditions have an important effect on the selection, survival, and growth of microorganisms
• Important environmental factors are

- Temperature
- pH
- Presence of toxic substances (i.e. heavy metals)
- Quality of the substrates (wastewater constituents)
• The environmental requirements for the bacteria vary with the species. E.g. chemo-autotrophic bacteria
are much more sensitive to environmental conditions than are heterotrophic bacteria
23
Environmental factors
• In general, optimal growth for a particular microorganism occurs within narrow ranges of temperature and pH
• Temperatures below the optimal level have a greater influence on the growth rate compared to temperatures
above the optimum
Type Temperature range Optimum range

°C °C
Psychrophilic 10 - 30 12 - 18
Mesophilic 20 - 50 25 - 40
Thermophilic 35 - 75 55 - 65
• The pH of the environment is also a key factor in the growth of microorganisms. Most bacteria cannot tolerate
pH levels above 9.5 or below 4.0
• Generally the optimum pH range for bacterial growth lies between 6.5 and 7.5
24
Fundamentals of microbial metabolism,

stoichiometry, process kinetics and biological
process models
- Stoichiometry of microbial reactions -
25
Stoichiometry of Microbial Reactions

Stoichiometry
Quantifies the relative quantities of reactants and products in chemical and biochemical reactions
here:
• Which amounts of material are to be used in the calculations performed for biological processes?
• Thereby, how can the different types of organisms be distinguished from one another?
26

General form of a process equation for a growth process:
Biomass Examples
+ substrate (energy source, electron donor)
+ electron acceptor O2, NO3-, CO2, org. substances,…

+ nutrients P, N, …
=
more biomass
+ metabolites H2O, CO2, …
+ products Org. substances
27

Cell composition and stoichiometry
• Composition of bacteria: ≈ 80% water; ≈ 20% dry material (90% of which is organic and ≈ 10% is inorganic)
• The most important chemical elements in organisms are:

C N P S H O
• The elemental composition of a bacteria cell the following proportions are typical:
C 51% 250 atoms

H 10% 611 atoms
O 21% 77 atoms
N 13% 55 atoms
P 3% 6 atoms
S 1% 1 atom
plus trace elements
28

Cell composition and stoichiometry
• The most widely used empirical formula for the organic fraction of cells is
C5H7O2N
• When phosphorus / sulfur are also considered:
C60H87O23N12P
• or
C250H611O77N55P6S
• these cumulative chemical formulations are approximations only and may vary with time and species
• all of these components should be obtained from the medium and the absence of any of them could
limit the growth of the bacterial population
29

Wastewater composition and stoichiometry
• Based on a „typical composition“ of domestic sewage, wastewater may be expressed with the formula
C500H1327O532N142P16S19
considering, however, only the compounds used by chemo-organo-heterotrophic bacteria
• These formulas are cumulative chemical formulas and enable a stoichiometric calculation of
transformations, occuring in cell metabolism
30

Some important metabolic process in wastewater treatment (anoxic and aerobic):
Wastewater Components Products

constituents coh cla -
bacteria bacteria O2 NO3 Me
COD X X Biomass
NH4+ X X NO3-
NO2-
NO3- X X N2
3- X X (PO43- )n in biomass
PO4
X Me PO43-
31

Degradation of organic carbon (aerated → aerobic conditions)

e.g. methanol: 2 CH3OH + 3 O2 ➔ 2 CO2 + 4 H2O
Nitrification (aerated → aerobic conditions)

2 NH4+ + 3 O2 ➔ 2 NO2- + 2 H2O + 4 H+ (nitritation)
2 NO2- + O2 ➔ 2 NO3- (nitratation)
Denitrification (not aerated → anoxic conditions)

5 CH3OH + 6 NO3- ➔ 5 CO2 + 7 H2O + 3 N2 + 6 OH-
32

Scheme of metabolism of chemo organo heterotrophic bacteria:
wastewater Syn- biomass

thesis
waste
decom waste
position
oxygen Energy waste

prod.
energy
33




5 CH3OH + 6 NO3- ➔ 5 CO2 + 7 H2O + 3 N2 + 6 OH-
34
Scheme of metabolism of chemo organo heterotrophic bacteria (Denitrification)
wastewater Syn- Biomass

thesis
waste
decompo waste
sition
Nitrate Energy
prod.
energy
35



5 CH3OH + 6 NO3- ➔ 5 CO2 + 7 H2O + 3 N2 + 6 OH-
36
Scheme of metabolism of chemo-litho autotrophic bacteria (nitrosomonas,

nitritation)
wastewater
Syn- biomass
thesis
waste
waste
oxygen Energy
prod.
energy
37
Scheme of metabolism of chemo-litho autotrophic bacteria

(nitrobacter/nitrospira, nitratation)
wastewater
Syn- biomass
thesis
waste
waste
oxygen Energy
prod.
energy
38
Fundamentals of microbial metabolism, stoichiometry,

process kinetics and biological process models
- Bacterial Growth and Biomass Yield -
39
Bacterial Growth and Biomass Yield

Biomass yield Y
• The ratio of the amount of biomass produced to the amount of substrate consumed is defined as
g biomass produced
Biomass yield Y =
g substrate utilized
• Y is typically defined relative to the electron donor. E.g.
Heterotrophic reactions with organic substrates:

g biomass / g organic substrate
Nitrification:
g biomass / g NH4-N oxidized
40

Example for a biomass yield: YH for aerobic heterotrophic growth
(see also Lecture 12 Wastewater Characteristics II)
Y: The ratio of the amount of biomass g biomass produced

produced to the amount of substrate Y=
consumed
g substrate utilized
biodegradable
organic
material
New cells
(COD produced)
Y
utilized by
heterotrophs 1
(biodegradable
O2 consumption
COD utilized)
(for energy
production)
1-Y
Typical values for heterotrophs : Y = 0.5 to 0.67 g COD of Biomass / g COD substrate
41

Yield coefficients of relevant microorgansims in wastewater treatment
• The biomass synthesis yield is related to the energy produced by the electron transfer
• The energy production that occurs from the oxidation reduction reactions is reduced as the electron acceptor is
changed from oxygen to nitrate to sulfate and CO2
• Moreover, conversion of CO2 to cellular carbon compounds requires more energy. Autotrophic bacteria must therefore
spend more of their energy for synthesis than do heteroptrophs
• These aspects are reflected in the synthesis of yield coefficient!

42

Estimating Biomass Yield and Oxygen Requirements from Stoichiometry
Example for the stoichiometric meaning of the biomass yield
Degradation of organic matter assumed to be in the form of glucose (C6H12O6)
New cells are assumed here to be represented by C5H7NO2
→ 3 C6 H12 O6 + 8 O2 + 2 NH3 2 C5H7NO2+ 8 CO2 + 14 H2 O

3(180) 8(32) 2(17) 2(113)
→ ΔC5H7NO2 2(113) g/mol

Y= = = 0.42 g cells / g glucose used
ΔC6 H12 O6 3(180) g/mol
43

Example for the stoichiometric meaning of the biomass yield
When COD is used to represent the organic matter and cells :
C6 H12 O6 + 6 O2 6 CO2 + 6 H2 O
180 6(32)
→ specific COD = 192/ 180 = 1.067 g O2 / g glucose

The theoretical yield expressed in terms of COD therefore is (with 1.42 g COD/g biomass VSS) (see example 3,
Wastewater Char. II) :
ΔC5H7NO2 2(113) g/mol 1 g glucose

Y= = = 0.39 g cells / g COD used
ΔC6 H12 O6 (3(180) g/mol ) 1.07 g COD
and
Y = (0.39 g cells / g COD used) 1.42 gCOD / g cells = 0.56 g COD / gCOD used
44

Solution to Example 3 (from Lecture 12):
C5H7NO2 + 5 O2 → 5 CO2 + NH3 + 2 H2O

C5H7NO2 : mw = 113 g/mol
O2 : mw = 32 g/mol
COD = 5 * 32/113 = 1.42 mg O2 / mg C5H7NO2
TOC = (5 * 12)/113 = 0.53 mg TOC / mg C5H7NO2
TOC 0.53 COD

→ = = 0.37 = 2.7
COD 1.42 TOC
Note: C5H7NO2 is often assumed to be a typical composition of biomass.
45

Example for the stoichiometric meaning of the biomass yield:
1 - Y = 1 – 0.56 = 0.44 g O2 / g COD used
46

Example for the stoichiometric meaning of the biomass yield:
The same result is obtained by using the stoichiometry directly (see slide 43):
8 (32) g O2
= 0.44 g O2 / g COD used
(3(180) g/mol ) 1.07 g COD/g glucose
or by a mass balance:
COD utilized = COD cells produced + COD of oxidized substrate
Oxygen consumed = COD utilized - COD cells produced
(3(180) g glucose/mol )1.07 g O2/g glucose - (2(113) g cells/mol) 1.42 g O2/g cells
= 577.8 - 320.9 = 256.9 g O2

256.9 g O2
= 0.44 g O2 / g COD used
(3(180) g/mol ) 1.07 gCOD/g glucose
47

process models
- Kinetics of microbial reactions -
48
Kinetics of microbial reactions

Definition
Kinetics: decription of the velocity at which a chemical reaction proceeds
= mathematical description
with consideration of the relevant

boundary conditions (e.g. temperature, pressure, concentrations
dc
Velocity: M L-3 T-1 kg m-3 h-1
dt
• The degradability of substrate and activity of the organisms determine the velocity at
which a substrate is degraded and its behavior
Both aspects together have to be considered
49

Some basic reaction types
Hypothesis The rate is directly dependent on the substrate concentration
rate = f ( cs )
with k :reaction constant
rate = k cs0 0 (zero) order reaction
rate = k cs1 1st order reaction
rate = k cs2 2nd order reaction
50

Graphical representation of the reaction order
rate Reaction First Order cs
-ks Cs1
Reaction Zero Order
-ks Cs0
cs t
rate – cs - Diagram c - t - Diagram
51

Enzyme-Catalyzed Reaction
Enzyme • chemically: protein
• Symbol: E
• substrate specific catalyst
Hypothesis Each reaction within a cell requires a specific enzyme
52
Enzyme-Catalyzed Reaction
• The metabolism processes within the organisms are understood as a chain of enzyme-catalyzed reactions
• Mathematically, enzyme-catalyzed reactions are usually described by a model based on the work of
Michaelis and Menten (1913)
• Reaction equation to describe the formation and decay of the enzyme-substrate-complex:
Principle
enzyme + substrate  enzyme-substrate-complex  enzyme + product
S Substrate
r1 r3
E + S ES E + P E Enzyme
r2 ES Enzyme-substrate-complex
P Product
r1,r2,r3 reaction rates
53
Single reactions and rate expressions

r1 r3
E + S ES E + P
r2
r1
E + S ES r1 = k1cEcS
cE , cS , cES =
concentrations [mol /L³];
r2
ES E + S r2 = k2cES k1 [L³ mol-1 T-1],k2,k3 [T-1]
= rate constants,
r3 r = partial reaction
ES E + P r3 = k3cES
54
r1 r3
Rate of change for the components E + S ES E + P
dcS r2
= -r1 + r2 = - k1cEcS + k2cES
dt
dcE
= -r1 + r2 + r3 = -k1cEcS + k2cES + k3cES
dt
dcES
= r1 - r2 - r3 = k1cEcS - k2cES - k3cES
dt
dcP
= r3 = k 3cES
dt
Also, the following balance holds: cE(t) = cE0 - cES (t)
This is a type of a non-linear ordinary differential equation system that can not be solved in a closed form.
However, …
55
Solution
…. under certain additional assumptions, the following expression is obtained
dcS k3 cE0 cS
- =
dt k2 + k 3
cS +
k1
k 3   k 1, k 2
k2 + k 3 k2 k2
≈ Km =
k1 k1 k1
dcS cS
with rmax = k3cE0
- = r = rmax and
dt cS + Km k
Km = 2
k1
56
Summary
Principle
enzyme + substrate  enzyme-substrate-complex  enzyme + product
Reaction equation
r1 r3
E + S ES E + P
r2
kinetic equation dcs cs

= k3 cE0
(Michaelis & Menten) dt k2
+ cs
k1
k 3   k 1, k 2
57
Monod Kinetics
Monod (1949) investigated the influence of the concentration of a dissolved nutrient on the velocity of
division of microorganisms and found a simple empirical mathematical way to describe the phenomenon
which is used until today:
dX 1 S Monod
+ = µ = μ max
dt X kinetics
Ks + S
This relation is very similar to the relation that Michaelis and Menten (1913) reported based on enzymatic
considerations.
Monod and Michaelis & Menten are therefore often confused with each other in literature.
Nevertheless, the theory of Michaelis & Menten and the resulting equations can describe the complex
reaction sequences, as they are present in organism cells, and are also related to the change of substances
outside the cell when certain assumptions are valid.
58

Kinetics of substrate utilization
• The theory of Michaelis & Menten and the resulting equations can be used to describe complex reaction
sequences in organism cells.
• However, certain assumption must be valid
• If these assumptions hold, the following relation results:
dS S
- = rSXmax X
dt S + KS
with
rSXmax = maximum substrate utilization rate related to biomass [M/(M T)]
KS = Half saturation constant [M/V]
S = Substrate concentration outside the organism [M/V]
X = Biomass concentration (all species X = Σ Xspecies j ) [M/V]
59

Kinetics of biomass growth
dS - dX dX S
- = = rSXmax YXS X
dt dS dt S + KS
with YXS = Yield coefficient [M/M]
dX 1 S
= rSXmax YXS
dt X S + KS
S
μ = μmax
S + KS
μ = growth rate [1/T]; μmax = maximum growth rate [1/T]
60

Resulting reaction equations
dS 1 S
dt = μ X
YXS max
Ks + S
rsx max
dX S µ : growth rate, T-1

+ = µX = μ max X
dt - dX
Ks + S = const = YXS
dS
S
r = µX = μ max X
Ks + S
61

Graphical representation
S S
S << KS ρ = μ max X ≈ μ max X
KS + S KS
S
S >> KS : ρ = μ max X ≈ μ max X
KS+ S
ρ
S→0
S→
μ max X
2
Ks S
62

process models
Model expressions for important

biological processes
63
Model expressions for important biological processes

Representation of biomass
• Biomass in the reactors are mixed cultures (unknown detailed composition)
• Therefore, biomass is described as a whole, without considering the detailed structure in the cell and
the detailed composition
• It is usually measured in pCOD or VSS
64

Representation of processes
• The mathematical description of the process rates depends on the properties of the substances used
(e.g., dissolved, colloidal, particulate) and the mechanisms in the process (e.g., inhibition, surface
limitation, …)
• The following expressions are examples of simple descriptions that have been proven to be of value
• However, there are many different formulations of empirical and theoretical nature in the literature that
may be used alternatively and lead to similar results!
• The expressions used, the substances incorporated, and the way the phenomena are described depend
on the situation and the question to be answered!
65

Growth of microorganisms
• The growth rate μ, depends on a number of factors such as temperature, pH, substrate- and nutrient concentration, toxicity,…
• Very often the empricial equation of Monod is used to describe the dependency of μ on the substrate concentration
ρ =μX
S
ρ = μmax X
KS + S
µ : growth rate, T-1
ρ = process rate [M L-³ T-1]
μmax = maximum growth rate at the given conditions (temperature, pH,..) [T-1]
KS= Half saturation coefficient for the substrate [MS L-³]
S = substrate concentration [MS L-³]
X = Biomass concentration [MX L-³]
66

Growth of microorganisms
• The value of μmax is e.g. dependend on the type of organism and on temperature
• Temperature dependency is often described by
μmax(T) = μmax(T0) exp[θ (T-T0)]
T = Temperature [°C]
T0 = reference Temperature [°C]
Θ = Temperature coefficient [°C-1]
μmax(T), μmax(T0) = μmax at T and T0
67

Decay and maintenance of microorganisms
• Decay processes describe a number of different processes that result in a loss of biomass. Therefore, decay is a
lumped process consisting of several single processes
• a living organism needs energy independently of growth, for
- renewing macromolecules (without synthesis of new cells)

- mobility
- maintaining inner structure and osmotic barriers
- etc.
• Under aerobic conditions the required energy is e.g. obtained through respiration of cell internal reserves (endogenous
respiration), this results in oxygen consumption independently of growth (reproduction)
• Additional losses of biomass result from predation by protozoa
68

Decay and maintenance of microorganisms
• Three different main concepts for the description of these processes may be distinguished:
- Endogenous Respiration
- Lysis
- Maintenance
• Each of these processes can describe phenomena regarding the decay and energy requirements of
biomass (all of them contribute to the whole „truth“). However, often just one process is chosen.
69

Endogenous Respiration:
Direct respiration of endogenous substances (cell internal reserves) to gain energy independently from growth /
reproduction
• Educts: Biomass, oxygen or an alternative electron donor

• Products: Metabolites (CO2, H2O, etc.), nutrients, inert particulate matter (nbpCOD)
• Direct oxygen consumption
- +
electron donor (e.g. O2)
-
Non biodegradable
biomass Endogenous
particulate matter
respiration
(nbpCOD)
nutrients
metabolites
70

Endogenous Respiration:
The rate of endogenous respiration is proportional to the biomass present
ρ∼ X

X = concentration of biomass [MX L-³]
bend = endogenous respiration rate [T-1]
The rate expression depends on the situation. If aerobic endogenous respiration is considered the rate may be
expressed as
O2
ρ = bend X
KO2 + O2
KO2 = half saturation constant for oxygen [M L-³ ]

O2 = dissovled oxygen concentration [M L-³]
to express that oxygen is needed
71

Lysis
• Break up of cells
• Educts: Biomass
• Products: Several products, e.g. biodegradable organic matter, and nonbiodegradable particulate matter (nbpCOD), nutrients
• No direct oxygen consumption, however, it results in indirect oxygen consumption through use of lysis products (bCOD)
non biodegradable
particulate organic
matter (nbpCOD)
biomass lysis Biodegradable

organic matter
- (bCOD)
nutrients
+
72

Lysis
The process is usually described by a first-order reaction:
ρ = blysis X

blysis = decay rate for lysis [T-1]
73

Maintenance
• Utilization of external substrate to gain energy independently from growth / reproduction (without synthesis of new cells)
• Educts: Substrate, oxygen
• Products: None, energy
• Direct oxygen consumption
electron donor (O2)

-
substrate Maintenance No products (only energy)
74

Maintenance
The process is proportional to the biomass
ρ∼ X

Oxygen and substrate are needed. The process rate may therefore e.g. be expressed as
O2 S
ρ = bmaint X
KO2 + O2 KS + S
KO2 = half saturation constant for oxygen [M L-³ ]

O2 = dissolved oxygen concentration [M L-³]
S = substrate concentration [MS L-³]
X = Biomass concentration [MX L-³]
bmaint = decay rate maintenance [T-1]
75

Hydrolysis of colloidal and particulate organic matter
• Usually, microorganisms can utilize only dissolved material (e.g. rbCOD)
• Exoenzymes (released in the external medium ) lead to hydrolysis reactions outside the cell, in the
liquid medium, converting large and complex substrate molecules into smaller and simpler molecules,
which can then pass through the cell membrane to become available for consumption by the cell.
Hydrolysis makes particulate or colloidal organic material available for microbial

metabolism
76

Hydrolysis
• Converts particulate matter into smaller compounds
• Educts: particulate matter
• Products: dissolved matter (sometimes also non-biodegr. may be included)
• No oxygen consumption
Dissolved non biodegradable organic

matter (nbsCOD)
Particulate Dissolved biodegradable organic

organic Hydrolysis matter (rbCOD)
matter
Dissolved nutrients
77

Hydrolysis
• Hydrolysis is a surface limited reaction and requires contact between heterotrophic organisms and particulate
substances. The rate can be described by a rate often used for surface limited reactions:
C/X
ρ = khyd X
KX + C/X

kH = hydrolysis rate constant [MC MX-1 T-1]
K = hydrolysis saturation constant [MC M-1]
C = concentration of particulate substances [MC L-³]
X = heterotrophic biomass [MX L-³]
• Hydrolysis is a process, that is characterized by the interrelation of many different factors, microorganism types and
different particulate and colloidal substances.
• It is a sum of several processes and the described equations allow only for a very simplified description and are mainly
of empirical nature.
78

Hydrolysis extreme cases Microorganisms (X)
Microorganisms (X)
C/X >> KX : The amount of microorganisms limits the reaction. The process rate
simplifies to:
C/X C/X
ρ = khyd X ≈ khyd X= khyd X
KX + C/X C/X
79

Hydrolysis extreme cases
Microorganisms (X) Microorganisms (X)
C/X << KX : The supply of particulate matter is the limiting factor. The process rate simplifies to:
C/X C/X khyd

ρ = khyd X ≈ khyd X= K C
KX + C/X KX X
80

Inhibition
• Microbiological processes are often inhibited by certain substances (i.e. run more slowly). These substances can be:
(1) Substances not involved in the reaction but being toxic or in competition with the reacting substances
(competitive inhibition)
(2) High concentrations of substrates (substrate inhibition)
(3) High concentrations of products of the reaction (product inhibition)

• It is usually described by:
KI
ρinhibited = ρnot inhibited
KI + Si
KI = Saturation coefficient for inhibition [M L-³]

Si = concentration of inhibiting substance [M L-³]
ρnot inhibited = reaction rate e.g. described by one of the expressions mentioned before
81
Note:
• The kinetic parameters (and especially the maximum growth rate µmax and the half saturation constants K)
are no unique parameters!
• They represent the net effect of certain factors of microbial kinetics on the simultaneous degradation of a
variety of different wastewater constitutents
• They depend on:
- conditions in the environment (temperature, pH, …), and

- the type and composition of the microbial population and the substrate
and, therefore have to be determined experimentally (or have to be calibrated/fitted for a certain situation)!
82
Values of kinetic and stoichiometric expressions in

different design procedures
procedure µN bA bH YN YH GV KN
[ 1/d ] [ 1/d ] [ 1/d ] [ gTS/gNH4 ] [ kg/kg ] [-] [ mg/l ]
EPA 0,288 − 0,06 − 0,7 oTS/BSB 0,6 1
Eckenfelder 0,276 0,034 0,068 0,2 TS/BSB 0,7 1 ) oTS/BSB 0,75 0,5
0,55 1 b) oTS/BSB
Sen 0,210 0,075 0,104 0,133 TS/CSB 0,45 2 ) 0,75 0,22

oTS/CSB
0,272 2 b) oTS/CSB
Denmark − − − − 0,7 - 0,8 TS/BSB − −
Japan − − 0,025 - 0,035 − 0,5 - 0,6 TS/BSB − −
Austria 0,342 0,050 0,180 −  − 1
Switzerland 0,29 − − −  − −
Barnard 0,202 0,050 0,180 − 0,45 oTS/CSB 0,83 −
WRC 0,113 0,03 0,180 0,10 0,45 oTS/CSB 0,83 0,31
A 131 0,319 3 ) 0,031 0,057 − 0,6  0,7 −
TS/BSB
Denika ‘89 0,319 0,032 0,056 0,15 TS/BSB 0,6  0,7 1
TS/BSB
range 0,11-0,34 0,03-0,075 0,025-0,18 − − 0,6-0,83 0,22 - 1,0
Wichern (1996) 83
Range of kinetic and stoichiometric parameters of

the design procedures
Souce: Wichern et al. (2004) 84

Examples for the use of kinetic expressions in the calculation of

the aerobic Sludge Age in design procedures for AS systems
pH influence
no decay rate
no decay rate
pH influence
Source: Wichern et al. 2004 85

Limitation by more than one substance
• In biological processes, a multitude of different substances are transformed that are needed as
reactants (substrate, nutrients, biomass)
• The Monod kinetics has turned out to be useful to describe the substrate limitation for microbial growth
in biochemical processes
• To describe that more than one substance is needed for a process to run, often multiplication of different
Monod terms is applied
• Although this multiplication is not based on biochemical fundamental considerations it has turned out to be
a useful possibility to describe kinetic relationships in a mathematically easy way
86

Example for a description of aerobic growth of heterotrophs as used in ASM1 (activated sludge model No 1, a
dynamic simulation model)
Metabolism works only if
substrate and electron acceptors are available:
S O2
Aerobic growth of heterotrophs ρ = μH,max XH
KS + S KO2 + O2
ρ = process rate [M L-³ T-1] electron donor electron

μH,max =maximum growth rate of heterotrophs [T-1] and carbon acceptor
KS , KO = Half saturation coefficients [M L-³] source
S, O2 = concentration of rbCOD, Oxygen [MS L-³]
X H= active heterotrophic biomass concentration [MX L-³]
87

Example for a description of anoxic growth of heterotrophs as used in ASM1 (activated sludge model No 1, a
dynamic simulation model)
Anoxic growth of heterotrophs (Denitrification)

S NO KO2
ρ = μH,max η XH
KS + S KNO + NO KO2 + O2
electron donor Electron Inhibition

and carbon acceptor by oxygen
source
μH,max =maximum growth rate of heterotrophs [T-1]
KS , KNO , KO = Half saturation coefficients [M L-³]
S, O2, NO = concentration of rbCOD, Oxygen, Nitrate and Nitrite [MS L-³]
X H= active heterotrophic biomass concentration [MX L-³]
η = correction factor for anoxic growth
88

process models
Process Model Equation Systems
89
Process Matrix Representation

Simplified example: Process equation:
• Growth of heterotrophic biomass 0 = -a Ss – (-b) O2 + XH
• Decay of heterotrophic biomass
• Aerobic conditions
-: represents used components
• No nitrification
+: represents generated components
Component i XH SS O2 Process rate rj
Process rate j
1 Growth 1 -1/YH 1-1/YH μmax (SS/(SS+KS)) XH
2 Decay -1 0 bH XH
Reaction rate
ri = Sr =Sv
ij ij rij
Substrate
Biomass
demand
Oxygen
90
Process Matrix Representation

Component i XH SS O2 Process rate rj
Process rate j
1 Growth 1 -1/YH 1-1/YH μmax (SS/(SS+KS)) XH
2 Decay -1 0 bH XH
Substrate
Biomass
demand
Oxygen
Biomass production dXH/dt = rXH = μmax (SS/(SS+KS)) XH - bH XH
Substrate utilization dSs/dt = rS = - 1/YH μmax (SS/(SS+KS)) XH
Oxygen demand dO2/dt = rO2 = (1-1/YH) μmax (SS/(SS+KS)) XH

91
SI
Soluble inert organic matter (nbsCOD) -1
Readily biodegradable substrate (rbCOD) YH

SS XI
Particulate inert organic matter (nbpCOD)
Slowly biodegradable substrate (sbCOD)

1
Active heterotrophic biomass

XS XH
Active autotrophic biomass

XA
Particulate products arising from biomass

XP
decay (nbpCOD)
Oxygen
SO
YH
-(1-YH)
O
Nitrate and nitrite

SN
NH4 and NH3 nitrogen

-ixB
SNH
Soluble biodegradable organic nitrogen

SND
Particulate biodegradable organic nitrogen

XND
Alkalinity
SALK
-ixB/14
Note: XI and XP may also be modelled together using one expression only
ρ = μH,max XH
Process rate
KS + S
O2
Example: The process matrix of the ASM1 (one row)
KO2 + O2
92
Example 1 for a simple dynamic model (not the ASM)

Heterotrophic growth in a batch reactor (e.g. BOD-test, neglecting nitrification)
Biodegradable COD is considered as fully readily biodegradable
dXH
= Biomass growth rate + (-1) Biomass decay rate
dt
dSS
= - (1/YH) Biomass growth rate + 0
dt
dXI
= 0 + (f) Biomass decay rate
dt
dO2
= (1-1/YH) Biomass growth rate + (-1+f) Biomass decay rate
dt
93
XH = Heterotrophic Biomass [mg COD/l];

SS = readily biodegradable organic substances (rbCOD) [mg COD/l]
O2 = dissolved Oxygen [mg O2/l];
SI = non-biodegradable soluble organic material (nbsCOD) [mg COD/l]
XI = non-biodegradable particulate organic material (nbpCOD) [mg COD/l]
YH = Biomass Yield of Heterotrophic Biomass;
f = Production of XI in the decay process
Matrix
Process rate XH SS XI O2 SI
μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH 1 -1/YH 0 1-1/YH 0
bH (O2/(O2+KO2)) XH -1 0 f f-1 0
94

Resulting ODE system (if no in - or outflow is considered)
dXH
= μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH - bH (O2/(O2+KO2)) XH
dt
dSS
= -(1/YH) μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH
dt
dXI
= f bH (O2/(O2+KO2)) XH
dt
dO2
= (1-1/YH) μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH + (f-1) bH (O2/(O2+KO2)) XH
dt
95
Example 1 for a simple dynamic model

Simulation results (organic material)
XI and SI
0
t
96
Example 1 for a simple dynamic model
Simulation results (Oxygen)
0 t
The term O2/(O2+KO2) could be neglected in the given simulation, since O2 is >> KO2
97
Accumulated organic material remaining (i.e. COD remaining)
O2
O2 consumed
rbCOD consumption until t
COD nbCOD
total biomass produced
until t
nbCO Biomass
D remaining
at t
0 t
98

Heterotrophic growth in a batch reactor (e.g. BOD-test, neglecting Nitrification)
Biodegradable COD is considered as rbCOD and sbCOD
dXH
= Biomass growth + (-1) Biomass decay + 0
dt
dSS
= (-1/YH) Biomass growth + 0 + Hydrolysis
dt
dXI
= 0 + (f) Biomass decay
dt
dXS
= 0 + 0 + (-1) Hydrolysis
dt
dO2
= (1-1/YH) Biomass growth + (-1+f) Biomass decay
dt
dSI
= 0
dt
99

XH = Heterotrophic Biomass [mg COD/l];
SS = readily biodegradable organic substances (rbCOD) [mg COD/l]
XS = slowly biodegradable organic substances (sbCOD) [mg COD/l]
O2 = dissolved Oxygen [mg O2/l];
SI = non-biodegradable soluble organic material (nbsCOD) [mg COD/l]
XI = non-biodegradable particulate organic material (nbpCOD) [mg COD/l]
YH = Biomass Yield of Heterotrophic Biomass;
f = Production of XI in the decay process
Process rate XH SS XI O2 XS SI
μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH 1 -1/YH 0 1-1/YH 0 0
bH (O2/(O2+KO2)) XH -1 0 f f-1 0 0
kHyd ((XS /XH)/(XS/XH +KX)) XH 0 1 0 0 -1 0
parameter values were not changed compared to the first model to show the effect of the new
process and COD fraction
100

Resulting ODE system (if no in- or outflow is considered)
dXH
= μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH + (-1) bH (O2/(O2+KO2)) XH
dt
dSS
= (-1/YH) μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH + kHyd ((XS /XH)/(XS/XH +KX)) XH
dt
dXI
= (f) bH (O2/(O2+KO2)) XH
dt
dX (-1) kHyd ((XS /XH)/(XS/XH +KX)) XH
S =
d
t
dO2
= (1-1/YH) μmax (SS/(SS+KS)) (O2/(O2+KO2)) XH + (f-1) bH (O2/(O2+KO2)) XH
dt
dSI
= 0
dt
101
Simulation results (organic material)
XI and SI
0 t
102

Simulation results (Oxygen)
0 t
The term O2/(O2+KO2) could be neglected in the given simulation, since O2 >> KO2
103

Accumulated organic material remaining (i.e. COD remaining)
O2
rbCOD
consumed
until t
sbCOD nbCOD
COD
total biomass produced
until t
nbCO Biomass
D remaining
t at t
0
104
Biological Nutrient Removal (BNR) Facilities
Dr.-Ing. Benedikt Aumeier

18 January 2024
1
The Activated Sludge Process
https://www.youtube.com/watch?v=R9R9jYUvgSI 2
Reactor Configurations – Complete mix
Source: Suez Degremont Handbook 3

Reactor Configurations – Plug-flow

Reactor Configurations – Step feed

Reactor Configurations – Ditch type

Two (2) Stage with Internal Recycle (IR)
Modified Ludzack-Ettinger (MLE) Process
Source: Hazen and Sawyer 2016 7

Anaerobic / Anoxic and Oxic (A 2/O) Process 3 Stage/1 IR

Anaerobic/anoxic and oxic (A2/O) Process
Sedlak: 1991
Source: Hazen and Sawyer 2016 8
Biological Nutrient Removal (BNR) Processes

• Modified Ludzack-Ettinger (MLE) Process – continuous-flow suspended-growth process with an initial anoxic stage followed by an
aerobic stage; used to remove TN
• A2/O Process – MLE process preceded by an initial anaerobic stage; used to remove both TN and TP
• Step Feed Process – alternating anoxic and aerobic stages; however, influent flow is split to several feed locations and the recycle
sludge stream is sent to the beginning of the process; used to remove TN
• Bardenpho Process (Four-Stage) – continuous-flow suspended-growth process with alternating anoxic/aerobic/anoxic/aerobic stages;
used to remove TN
• Modified Bardenpho Process – Bardenpho process with addition of an initial anaerobic zone; used to remove both TN and TP
• Sequencing Batch Reactor (SBR) Process – suspended-growth batch process sequenced to simulate the four-stage process; used to
remove TN (TP removal is inconsistent)
• Modified University of Cape Town (UCT) Process – A2/O Process with a second anoxic stage where the internal nitrate recycle is
returned; used to remove both TN and TP
• Rotating Biological Contactor (RBC) Process – continuous-flow process using RBCs with sequential anoxic/aerobic stages; used to
remove TN
• Oxidation Ditch – continuous-flow process using looped channels to create time sequenced anoxic, aerobic, and anaerobic zones; used
to remove both TN and TP.
9
Average of COD and NH4-N Effluent Concentrations

of WWTPs across Germany
DWA (2018). Leistungsvergleich 2017 10

Average of Total N and Total P Effluent

Concentrations of WWTPs across Germany

Average Influent and Effluent Concentrations and

Performance of WWTPs across Germany

Activated Sludge Processes I

- Modeling Suspended and Attached Growth Systems

25 January 2024

1
Suspended and attached growth

Suspended growth treatment processes:
• The active biomass is suspended in and transported with the liquid phase
• The technologies that use suspended biomass mimic natural systems where biomass is suspended
and transport through advection, turbulence, and others dominates (e.g., lakes, lagoons, rivers)
Attached growth treatment processes:

• The active biomass is attached to a surface and the water flows
relative to the biomass
• Biofilm technologies mimic natural systems where the attached biomass dominates the processes (e.g.,
in shallow rivers)
2

Suspended growth treatment processes
Example: Activated sludge system
3

Bacterial - Floc
(green: EPS , red: Bacteria)
CLSM image CLSM image
Source: Christian Staudt, Thomas Neu, UFZ, Department Fließgewässerökologie 4


Attached growth treatment processes
Example: Trickling filter
5

Biofilm
150
100
50
0
z [µm]
-50
-100
-150
-200
100 µm -250
-300
0 5 10
Oxygen concentration
[mg/l]
6

Biofilm
Biofilm 3 D
Area: 500 x 500 µm
7
Mass balances for dynamic and steady-state conditions

Important (ideal) reactor types
Batch reactor:
• No inflow
• No outflow
Inflow
Ideal continuous completely stirred tank reactor
(CSTR) (Durchlauffermenter):
• with suspended biomass Outflow
Plug Flow reactor:

Inflow Outflow
8
Mass balances for dynamic and steady state conditions

Important reactor types
Inflow
Activated sludge system:
• Reactor with suspended biomass and organism
recycling
Sludge Recycling
Biofilm-fixed bed-reactor: Inflow
Outflow
9

Ideal continuous flow completely stirred tank
with suspended biomass

Q,
Outflow
Cin
Q, Cout
biological reactions ri
C Inflow
C is everywhere the same in the reactor Czu
and Creactor = Cout
Reactor Outflow
C Cout
Location
10

Mass balance of a CSTR
Assumption: Perfect Mixing (CSTR, Completely stirred tank reactor)
System boundary
Qi
n
Ci
n
Qout
Cout
V
C
11

General written statement for a mass balance :
Time variation of Mass of the Mass of the Mass of the

the mass of the = component that - component that component
+-
component in the goes into the leaves the system produced or
system system per time per time consumed by
reactions per time
accumulation = inflow - outflow + generation

(Positive or negative sign)
The input and output terms are transport terms and depend upon the physical
characteristics of the system being modeled (and the reactor type)
12

The Notion of Mass balances
General written statement for a CSTR :
Time variation of Mass of the Mass of the Mass of the

the mass of the = component that - component that component
+-
component in the goes into the tank leaves the tank produced or
tank per time per time consumed by
reactions per time
d(VC)
dt = Σ Qin Cin – Σ Qout Cout + V ri
13

Mass balance of a CSTR
d(VCi)
dt = Qin Ci,in – Qout Ci,out + V ri
dCi dV
V + Ci = Qin Ci,in – Qout Ci,out + V ri
dt dt
If the volume does not change:

dCi Qin Qou
dt = Ci,in – t Ci,out + ri
V V ri: reaction rate of component i (sum of reactions or
net reaction rate if there are more reactions) [M / L³ T]
transport conversion V: Volume [L³]
By defining the dilution rate D=Qin/V : Qin,Qout : flow [L³ / T]
dCi Cin, Cout, C: Concentrations of component i [M/L³]

= D Ci,in – D Ci,out + ri
dt
14

Steady-state simplification
If only the steady-state (long-term) concentration is of interest :
accumulation = inflow + outflow + generation
=0
0 = inflow + outflow + generation
15

Mass balances for conservatives
Examples for conservatives gas N2
(in an aerated tank) System boundary
influent Biological effluent

processes
N P N
COD or ThoD*
COD or
ThoD* P
*ThoD (Theoretical Oxygen demand) is the

conservative form of COD (COD analysis input waste N
recovers only a fraction of the ThOD, typically O2 (sludge) COD or ThoD*
95% in domestic wastewater) (negative P
COD or ThOD)
16

Mass balances for conservatives
Examples for conservatives gas N2
System boundary
(in an aerated tank)
influent Biological effluent

processe
Flow: Qin Flow: Qout
s V
Substrate: Sin Substrate: Sout
XB,H, XD
input waste
O2 (sludge) Flow: QW
17

Continuity in the processes for conservatives
Processes j Components i Process
equations
Component 1 Component 2 Component 3 Component n
1
2 ʋ ji ρj
3
m
ri
ρj : Process equation for process j m m

ri : summed up reaction equation for component i ri = Σ j=1 rji = Σj=1 ʋji ρ j
ʋji : stoichiometric constant
The continuity can be checked by moving across the matrix
Providing consistent units have been used, the sum of the stoichiometric coefficients must be zero
! 18

Continuity: Example for a simple kinetic model
(heterotrophic bacteria growth in an aerobic environment)
Process Components Process

equations
Substrate Nutrient Het. Oxygen Non biodegr.
ρ
S SNH4-N Biomass O2 part. org. matter
X XI
(rbCOD, (NH4 (het. (O2, (measured as

measured measured organisms negative COD)
as COD) as N) measured COD)
as COD)
growth - 1 / YH -iN,BM 1 1-1/YH µmax (S/(S+KS))X
decay iN,BM - -1 -(1-fXI) fXI bX
iN,XI*fXI
fXI : Portion that generates XI in endogenous respiration (port. of inert org. products) [gCOD/gCOD]
iN,BM : N content of biomass [gN / gCOD]
iN,XI : N content of XI [gN / gCOD] à See also example 4 in Lecture 12 (but rel. to COD)
YH : Aerobic synthesis Yield of heterotrophic biomass [g COD / g COD]
19

Continuity: Example for a simple model
(heterotrophic bacteria growth in an aerobic environment)
Process Components Process
equations
Substrate Nutrient Het. Oxygen Non biodegr.
ρ
S SNH4-N Biomass O2 part. org. matter
X XI
(rbCOD, (NH4 (het. (O2, (measured in

measured measured organisms negative COD)
as COD) as N) measured COD)
as COD)
growth - 1 / YH -iN,BM 1 1-1/YH µmax (S/(S+KS))X
decay iN,BM - -1 -(1-fXI) fXI bX
iN,XI*fXI
COD in growth : 1*(-1/YH) + 1*(1) +(-1)*(1-1 / YH) = 0
COD in decay : 1*(-1) + (-1)*(-(1-fXI)) + 1*fXI = 0
N in growth : (1)*(-iN,BM) + iN,BM*(1) = 0
N in decay : (1)*(iN,BM-iN,XI*fXI) + iN,BM*(-1) + iN,XI* fXI = 0
20
Continuous flow systems with suspended biomass (a simple AS system)

A simple activated sludge system
Activated sludge tank

oxygen
Secondary clarifier
Inflow (prim. effl.)
effluent
Excess sludge
Return sludge
21

A simple activated sludge system
oxygen
Q
Q+QR
Q-QW
XT,0 XT,e Ce Se
C0 V
XT C S QW+QR
S0
QW
QR XT,R CR SR XT,w Cw Sw
XT : Total suspended solids concentration
C : concentration of pollutants (dissolved and particulate)
22

Mass balance around the aeration tank
oxygen System
boundary
Q
Q+QR
XT,0
C0 V
S0 XT
CS
QR
SR CR XT,R
V dX
dt = Q XT,0 – (Q+QR ) XT + QR XT,R + rXT V
T XT : Total suspended solids concentration
V dC
dt = Q C0 – (Q+QR ) C + QR CR + rC V
C : concentration of pollutants (dissolved and particulate)
23

Mass balance around the secondary clarifier
• It is assumed that there are no biological reactions in the secondary clarifier
• The sludge blanket level in the secondary clarifier is assumed to remain constant, i.e. the storage of
XT in the secondary clarifier is negligible System
boundary
XT (Q+QR)
(Q-QW) XT,e
(QW+QR) XT,w
QW XT,w
QR XT,R
0 = (Q+QR) XT – (Q-QW) XT,e – (QR+QW) XT,W
24

Mass balance around the whole system
System
boundary
Q XT
(Q-QW) XT,e
XT,0
QW XT,w
V dX
dt = Q XT,0 – [(Q-QW ) XT,e + QW XT,R ] + rXT V
T XT : Total suspended solids concentration
V dC
dt = Q C0 – [(Q-QW ) Ce + QW CT,R ] + rC V
C : concentration of pollutants (dissolved
and particulate)
25

Total suspended solids Mass Balance (alternative view)
V dX
dt = Q XT,0 – (Q+QR ) XT + QR XT,R + rXT V
T
0 = (Q+QR) XT – (Q-QW) XT,e – (QR+QW) XT,W
0 +V dXT = (Q+QR) XT – (Q-QW) XT,e – (QR+QW) XT,W +Q XT,0 – (Q+QR ) XT + QR XT,R + rXT V
dt
dXT = – (Q-Q ) X – (Q ) X +Q X + r V Which is the same expression as obtained with the total mass
à dt W T,e W T,W T,0 XT balance (XT,R = XT,w )
26

Total suspended solids mass balance
Many models of the simple activated sludge process assume that
• the amount of sludge in the secondary clarifier is negligible,
• there are no biological reactions in the secondary clarifier,
• the volume of the secondary clarifier is negligible
With these assumptions, the secondary clarifier does not appear in the mass balance:
V dX
T
With the assumption that the concentration of biomass in the influent is negligible (XT,0 = 0) :
1 dXT (Q-QW ) XT,e + QW XT,R rXT

= +
XT dt V XT XT
Definitions:
XT : Total suspended solids concentration
(Q-QW ) XT,e + QW XT,R DX : Dilution rate of XT
DX =
V XT
μn : net production rate of XT
rXT
µn =
XT 27

With
V dX
T
1 dXT = D + µ
X n
XT dt
The operation of the AS system often aims at keeping the concentration of XT constant by adjusting QT,W * XT,w to the
production of XT,w in the system.
Over long periods this gives:
∫ (1/XT) (dXT / dt) dt = 0

∫ DX dt = ∫ µn dt
or
DX ,mean = µn, mean
28

DX determines whether, a specific organism group can exist in the system
Therefore, DX (or more often its inverse value SRT) is considered the most important design
parameter for activated sludge systems
1/DX = SRT (solids retention time or sludge age)
Solids mass in the reactor

SRT =
Solids removed from system
29

Hydraulic retention time 𝝉 vs. solids retention time (SRT)
Systems with sludge recycling 𝝉 < SRT
Systems without sludge recycling

𝝉 = SRT
1 Volume of liquid in the system

= HRT = 𝝉 =
Dh Volume of liquid removed from the system
1 Solids mass in the reactor

DX = SRT =
Solids removed from system
30

Effect of SRT on the performance of an activated sludge system
(red Substrate, blue Biomass)
gVSS/m³
gCOD/m³
gCOD/m³
Wash out Wash out

Values used:
Si ∞= KS (1+kd (1/DX) )/((1/DX) (µmax-kd)-1) Y=0.4 gVSS/gCOD
μmax = 5 gVSS/gVSS d
X∞ = (Dh/Dx)Y (Si,in – Si,∞ )/(1+kd*(1/DX) ) kd = 0.1 gVSS/gVSSd
Ks = 10 gCOD/m³
Sin Substrate (bsCOD) Sin =192 gCOD/m³
wash out at DX >= µmax - Xin = 0
S in+ KS kd V = 197 m³
Q = 1000 m³/d
31

(red Substrate, blue Biomass)
gVSS/m³
gVSS/m³
gCOD/m³
gCOD/m³
Wash out Wash out

Values used:
Y=0.4 gVSS/gCOD
Si ∞= KS (1+kd (SRT) )/((SRT) (µmax-kd)-1) μmax = 5 gVSS/gVSS d
kd = 0.1 gVSS/gVSSd
X∞ = (SRT/𝝉)Y (Si,in – Si,∞ )/(1+kd*(SRT) )
Ks = 10 gCOD/m³
Substrate (bsCOD) Sin =192 gCOD/m³
Xin = 0
V = 197 m³
Q = 1,000 m³/d
32
Simple activated sludge system, design equations using simple rate expressions
Terminology used in the following expressions

• In the following formulation of kinetic expressions biodegradable soluble COD (bsCOD) will be used to
quantify the fate of biodegradable organic compounds because it relates easily to the stoichiometry of
substrate oxidized or used in cell growth
• For complete-mixed suspended growth designs if the SRT is 3 d or more, essentially all of the
degradable particulate COD will be converted to bsCOD
• The biomass solids in a bioreactor are commonly measured as total suspended solids (TSS) and
volatile suspended solids (VSS). The mixture of solids resulting from combining recycled sludge with
influent wastewater in the bioreactor is termed mixed liquor suspended solids (MLSS) and mixed liquor
volatile suspended solids (MLVSS)
• Besides the organic (volatile) fractions of solids the influent wastewater contains inorganic total
suspended solids (iTSS or termed FSS in former lectures)
33
Mass balances for soluble substrate and biomass
dS = Q S – QS + r V
V dt 0 SU
V dX
dt = Q X0 – [(Q-QW ) Xe + QW XR ] + rg V
where
dX
= rate of change of biomass concentration in the reactor measured as gVSS/m³ d
dt
V : reactor volume (i.e. aeration tank), [m³]
Q : influent flowrate, [m³/d]
X0 : concentration of biomass in influent, [g VSS/m³]
S0 : influent soluble substrate concentration, [g/m³]
QW : waste sludge flowrate [m³/d]
Xe : concentration of biomass in effluent, [gVSS/m³]
XR : concentration of biomass in return line from clarifier [gVSS/m³]
rg : net rate of biomass production, [ gVSS/m³ d]
rSU : rate of substrate concentration change due to utilization, [g/m³ d]
34
Kinetic Process Matrix However, other
The simple process matrix below can be used to describe the aerobic system (X in the matrix is expressions may
described in COD units): be equally valid
Process Soluble Biomass O2 Xinert

Substrates
growth - 1/Y 1 -1/Y+1 µmax S
X
S + KS
decay -1 fd-1 fd kd X
Note: The soluble substrate can e.g. be

organic substances if X are heterotrophs
or e.g. ammonia, nitrite if X are autotrophs
α=1 for heterotrophic growth
Xinert : non biodegradable part. org. matter

35
Ranges of kinetic coefficients often used (VSS based)
µmax/Y
Remember: the kinetic parameters are not unique and may change
depending on the specific loal condition!
36
Rate of Utilization of Soluble Substrates Corresponds to:
37
Net production rate of biomass Corresponds to:
μ is defined here such that it corresponds to a

net growth rate of growth - decay
When biomass is expressed in VSS :
38
Corresponds to:
Rate of Oxygen Uptake
However, the term f kd is not

included in the equation for ro
(given earlier in example Biomass is expressed here in VSS

in former lecture, and in
lecture notes)
(defined earlier)
(defined earlier)
39
Rate of nbVSS production from cell debris
Corresponds to:
40
Important Yield Expressions

Yield Expressions
Besides the synthesis yield two other yield terms are important in the design and analysis of biological
treatment processes:
• Synthesis Yield (or true Yield)

The amount of biomass produced during cell synthesis relative to the amount of substrate degraded
• Net Biomass Yield

The ratio of the net biomass growth rate to the substrate utilization rate
• Observed Yield
Accounts for the actual solids production that would be measured for the system
41

Synthesis Yield (defined in earlier lectures)
The amount of biomass produced during cell synthesis relative to the amount of substrate degraded
• Typically defined relative to the electron donor
• affected by
the energy that can be derived from the oxidation reduction reaction,
by the growth characteristics of the carbon source,
by the nitrogen source, and
by environmental factors such as temperature, pH, and osmotic pressure.
42

Net Biomass Yield
The net biomass yield is the ratio of the net biomass growth rate to the substrate utilization rate
(defined earlier)
(defined earlier)
43

Observed Yield
The observed yield accounts for the actual solids production that would be measured for the system (calculated as
gTSS/g bsCOD or gBOD, or as gVSS/g bsCOD or g BOD)
(defined earlier)
(defined earlier)
44

Observed Yield
VSS production PX,VSS

Yobs =. =
Substrate removed Q(S0-S)
(defined earlier)
if there is no nbVSS in the influent
45

Observed yield vs synthesis yield
Synthesis yield (or true yield) Observed yield
• amount of biomass produced • Includes also cell loss concurrent with cell
immediately upon consumption of the growth
growth substrate or oxidation of the electron
donor
• takes into account only the biomass • Contains other organic solids from the
(cells) produced wastewater that are measured as VSS, but
are not biologically degradable
• Seldom measured directly and often • Based on actual measurements of

interpreted frome evaluating biomass biomass production and substrate
production data for reactors operating under consumption
different conditions or based on theoretical
considerations
46
The simple activated sludge system, design equations using simple rate expressions
Biomass Mass Balance
System
boundary
Q X0 rg V
(Q- Xe
QW)
VX
QW Xw
47
Biomass Mass Balance
48
Biomass mass balance and solids retention time
At steady state and with X0 = 0 :
and
VX
= (defined earlier)
rg V
49
Substrate effluent concentration
50
Substrate Mass Balance
System
boundary
Q S0 rSU V
(Q- Se
QW)
VS
QW Sw
51
Substrate Mass Balance and Biomass concentration
At steady state :
and with
52
Mass Balance of nbVSS
V dXi = Q Xi,0 – [(Q-QW ) Xi,e + QW Xi,R ] + rX,i V

dt
System
boundary
Q Xi,0 rX,i V
(Q- Xi,e
QW)
V Xi
QW Xi,w
53
Mass Balance of nbVSS

dt
At steady state :
0 = Q Xi,0 – V Xi + fd kd X V because
SR
T (and SRT is valid for all fractions of the mixed liquor)
(given earlier)
54
Total Volatile Suspended Solids in the tank
• In general :
total VSS = biodegradable VSS + non biodegradable VSS

total VSS = bVSS + nbVSS
• The biomass in the aeration tank is measured here as VSS
• The total VSS in the aeration tank equals the biomass concentration plus the nbVSS concentration:
total VSS in the tank = biomass + nbVSS
• The kinetic expressions used to describe the biological processes are related to the active biomass
concentration (microorganisms) in the treatment reactor (X).
• the fraction of active biomass can vary depending on the wastewater characteristics and operating
conditions.
55
Mixed liquor suspended solids (MLSS) and Mixed liquor volatile suspended solids (MLVSS)
• The mixture of solids resulting from combining recycled sludge with influent wastewater in the bioreactor is termed
MLSS and MLVSS, respectively.
Biomass + nonbiodegradable Volatile Suspended Solids nbVSS + inert inorganic total suspended solids (iTSS or FSS)
MLSS = Biomass + nbVSS + FSS

MLVSS = Biomass + nbVSS
MLSS = VSS in the tank + FSS in the tank

MLVSS = VSS in the tank
56
Mixed liquor volatile suspended solids concentration (MLVSS)
• The total MLVSS in the aeration tank equals the biomass concentration plus the non biodegradable VSS (nbVSS)
concentration:
XT = X + Xi
With the expressions for X and Xi (given earlier):
57
58
Production rate of total VSS in the tank
total VSS in the tank = biomass + nbVSS
or XT = X + Xinert
and d(VSS) = d(biomass) + d(nbVSS)

d d d
t t t
or dXT dX dXinert
dt = dt + dt
with
dt
and
àV dX T = – [(Q-QW ) Xi,e + QW Xi,R ] – [(Q-QW ) Xe + QW XR ] + Q Xi,0 + rX,i V + rg V

dt
Production 59
Production rate of total VSS in the tank
With the expressions for rg and rXi (given earlier), and the nbVSS in the influent:
fd may vary from 0.1 to 0.15 gVSS/gVSS
= rXi
60
Solids Production
The amount of VSS produced and wasted daily is determined as follows:
Note: 1/SRT represents the fraction of solids wasted per day

Because the mixed liquor is a homogeneous mixture, the expression can be
used to calculated the amount of solids wasted of any of the mixed liquor
components
PX component * SRT = V*Xcomponent
with
(given earlier)
à PXT,VSS
61
Solids Production
with (given earlier)
62
Active Biomass
because: X/XT = PX / PXT,VSS
(defined earlier)
(defined earlier)
(defined earlier)
63
Solids Production
To calculate the solids production in terms of TSS the influent inorganic solids are added and the biomass is calculated in
terms of TSS by assuming a VSS/TSS ratio of 0.85:
However, the VSS/TSS ratio may vary from 0.8 to 0.9
64
MLSS, MLVSS and aeration tank volume
(given earlier)
This equation allows for a
determination of the aeration volume
by selecting an appropriate
MLSS concentration in the range of 2,000 to 4,000 mg/L
(must be compatible with the sludge settling characteristics and the clarifier design)
65
COD mass balance and oxygen requirements
Oxygen used = CODin - CODout
oxygen System
boundary
Q
(Q-QW) CODeffluent
CODinfluent
QW CODwaste
66
PX,bio includes active biomass and cell debris from cell growth and is the sum of terms A and B in:
(given earlier)
67
Design and Operating Parameters

Important design and operating parameters for activated sludge systems
• Solids retention time (SRT) German: „Schlammalter “

• Food to microorganism ratio (F/M) German: „Schlammbelastung BTS“
• Volumetric (organic) loading rate (Lorg)
68

Food to microorganism ratio (F/M)
69

F/M ratio and Efficiency
The F/M ratio can be related to the specific substrate utilization rate by the process efficiency :
U: Specific Substrate Utilization rate
70

F/M ratio, SRT, and Efficiency
Furthermore with and :
and
And with the relation between U, F/M and E :
71

Discussion of the F/M ratio
• The F/M ratio is historically the most important design parameter to design simple activated sludge systems.
• However, the F/M ratio is based on a highly simplified consideration of the process.
• Effects of loading variations e.g. over a day are only considered by experience.
• The F/M ratio is problematic if more complex processes are to be designed (e.g. if chemical elimination of P is
included, a big portion of the return sludge consists of precipitates, changing the relation between process performance
and F/M ratio.
• The F/M ratio only considers the BOD and is therefore not suitable if other aspects besides the BOD elimination
have to be considered
72

• High sludge ages are associated with low F/M ratios, and vice versa
• High F/M ratio: can mean a larger offer of biodegradable organic matter than the consumption capacity of the biomass
in the system, resulting in surplus substrate in the final effluent
• Low F/M ratio: can mean that the substrate offer is lower than the microorganisms‘ capacity to use it in the activated
sludge system. As a consequence, they will consume practically all the organic matter from the influent wastewater,
as well as their own cellular material
73

Organic Volumetric Loading rate (Lorg)
74

Solids Retention Time (SRT)
• The SRT is more suitable than the F/M ratio especially when Nitrification and Denitrification is to be incorporated. The
SRT is based on the growth rate of microorganisms in the aerated tank independently of the type of e.g. P-Elimination
• The SRT is based on a steady state consideration and effects of loading variations e.g. over a day are only
considered by experience
• For given values of the kinetic coefficients, the effluent substrate concentration from the reactor is a direct function of
the SRT. Setting the SRT value fixes the values of U and μ and also defines the efficiency of biological waste
stabilization
75

Process Performance and Stability
• The critical SRT at which the cells are washed out of the system is the minimum SRT SRTmin. When washout occurs S = S0.
• Using the simple expressions introduced earlier SRTmin is given by :
If S0 is much greater than KS (which is often encountered in wastewater treatment) :
To ensure adequate treatment and avoid system failure, biological treatment processes are operated with a design SRT
value from 2 to 20 times SRTmin
à Corresponds to a Safety Factor SF = SRTdesign/SRTmin
76
Activated Sludge Processes II

- Modeling Suspended Growth Systems for Biological Nutrient
Removal (BNR)

1 February 2024
1
Biological Nitrogen Removal
2
Example for a single sludge suspended growth system

Example for a single sludge suspended growth system:
• Bacteria: Aerobic autotrophic bacteria in suspended or attached growth systems
3
Example for a two sludge suspended growth system
effluent
• In cases where there is a significant potential for toxic and inhibitory substances in the
wastewater,
• First aeration tank/clarifier unit: Operated at a short SRT for BOD removal.
Removal of BOD and toxic substances
• Second unit:
Nitrification can proceed unhindered.
4
Example for a two sludge suspended growth system

WWTP Munich II – Gut Marienhof
Born and Ermel, 2022 5

Example for the concentration change along the flow axis in an

attached growth reactor (nitrifying trickling filter)
Nitrification starts only,
when COD degradation is lower Competition for oxygen !
concentration
àmost of the BOD must be removed

before nitrifying organisms can be
established
The heterotrophic bacteria dominate

the surface area of fixed-film systems
over nitrifiying bacteria
NH4-N COD NO3-N

distance
à Nitrifying trickling filters are often designed with 50% of the load (resulting in a
doubling of the volume) and the upper and lower part may be designed separately
Nitrification – Stoichiometry (suspended or attached)

The two-step process is involving two groups of bacteria:
Nitrosomonas: Ammonia à nitrite;
Nitrospira, nitrobacter: Nitrite à Nitrate
• The stoichiometry of the growth of nitrifiers is based on

- the conservation of COD (1 g NO3--N corresponds to - 4.57 g COD) - and the conservation
of nitrogen
• The yield coefficient of the nitrifiers is often defined as the nitrifiers produced per nitrogen oxidized to nitrate
7
Nitrification – Dominant Bacteria

ARTICLE doi:10.1038/nature16461 LETTER doi:10.1038/nature16459
Complete nitrification by Nitrospira Complete nitrification by a single microorganism

bacteria Maartje A. H. J. van Kessel1, Daan R. Speth1, Mads Albertsen2, Per H. Nielsen2, Huub J. M. Op den Camp1, Boran Kartal1,3,
Mike S. M. Jetten1,4 & Sebastian Lücker1
Holger Daims1, Elena V. Lebedeva2, Petra Pjevac1, Ping Han1, Craig Herbold1, Mads Albertsen3, Nico Jehmlich4,
Marton Palatinszky1, Julia Vierheilig1, Alexandr Bulaev2, Rasmus H. Kirkegaard3, Martin von Bergen4,5, Thomas Rattei6,
Nitrification is a two-step process where ammonia is first oxidized removed ammonium and nitrite from the medium (Extended Data
Bernd Bendinger7, Per H. Nielsen3 & Michael Wagner1 to nitrite by ammonia-oxidizing bacteria and/or archaea, and Fig. 1). The culture showed anaerobic ammonium-oxidizing (anam-
subsequently to nitrate by nitrite-oxidizing bacteria. Already mox) activity (Fig. 1a), and fluorescence in situ hybridization (FISH)
described by Winogradsky in 18901, this division of labour between revealed that anammox organisms of the genus Brocadia constituted
the two functional groups is a generally accepted characteristic of the approximately 45% of all FISH-detectable bacteria. Surprisingly,
Nitrification, the oxidation of ammonia via nitrite to nitrate, has always been considered to be a two-step process catalysed biogeochemical nitrogen cycle2. Complete oxidation of ammonia to Nitrospira-like nitrite-oxidizing bacteria accounted for approximately
by chemolithoautotrophic microorganisms oxidizing either ammonia or nitrite. No known nitrifier carries out both nitrate in one organism (complete ammonia oxidation; comammox) 15% of the community and co-occurred with the Brocadia species in
steps, although complete nitrification should be energetically advantageous. This functional separation has puzzled is energetically feasible, and it was postulated that this process flocs (Fig. 2a). This tight clustering with anammox bacteria was unex-
microbiologists for a century. Here we report on the discovery and cultivation of a completely nitrifying bacterium could occur under conditions selecting for species with lower pected as both microorganisms require nitrite for growth. Together
from the genus Nitrospira, a globally distributed group of nitrite oxidizers. The genome of this chemolithoautotrophic growth rates but higher growth yields than canonical ammonia- with the presence of Nitrospira at very low oxygen concentrations, this
organism encodes the pathways both for ammonia and nitrite oxidation, which are concomitantly expressed during oxidizing microorganisms3. Still, organisms catalysing this process indicated that there could be a functional link between these organisms.
growth by ammonia oxidation to nitrate. Genes affiliated with the phylogenetically distinct ammonia monooxygenase and have not yet been discovered. Here we report the enrichment and To determine the function of Nitrospira in the community, we
hydroxylamine dehydrogenase genes of Nitrospira are present in many environments and were retrieved on Nitrospira- initial characterization of two Nitrospira species that encode all extracted and sequenced total DNA from the enrichment culture bio-
contigs in new metagenomes from engineered systems. These findings fundamentally change our picture of nitrification the enzymes necessary for ammonia oxidation via nitrite to nitrate mass. In total 4.95 gigabase pairs of trimmed metagenomic sequence
and point to completely nitrifying Nitrospira as key components of nitrogen-cycling microbial communities. in their genomes, and indeed completely oxidize ammonium to were obtained and used for de novo assembly. By differential coverage
nitrate to conserve energy. Their ammonia monooxygenase (AMO) and sequence composition-based binning5 it was possible to extract
enzymes are phylogenetically distinct from currently identified high-quality draft genomes of two Nitrospira species. The two strains
Nitrification is catalysed by ammonia-oxidizing bacteria (AOB)1 The globally distributed genus Nitrospira represents the most AMOs, rendering recent acquisition by horizontal gene transfer had genomic pairwise average nucleotide identities (ANI)6 of 75%
or archaea (AOA)2 and nitrite-oxidizing bacteria (NOB)1. Since diverse known group of NOB. Nitrospira members have been found from known ammonia-oxidizing microorganisms unlikely. We and thus clearly represented different species (Nitrospira sp.1 and sp.2,
the pioneering studies by Winogradsky more than a century ago3, in terrestrial8 and limnic habitats 9,10, marine waters11, deep sea also found highly similar amoA sequences (encoding the AMO Extended Data Fig. 2 and Extended Data Table 1). Surprisingly, both
nitrifying microorganisms are generally perceived as specialized sediments, sponge tissue12, geothermal springs13, drinking water subunit A) in public sequence databases, which were apparently genomes contained the full set of AMO and hydroxylamine dehy-
chemolithoautotrophs that obtain energy for growth by oxidizing distribution systems14, corroded iron pipes15, and wastewater treat- misclassified as methane monooxygenases. This recognition of a drogenase (HAO) genes for ammonia oxidation, in addition to the
either ammonia or nitrite. The known ammonia-oxidizing microbes ment plants (WWTPs)10,16. At least six phylogenetic sublineages of novel amoA sequence group will lead to an improved understanding nitrite oxidoreductase (NXR) subunits necessary for nitrite oxidation
(AOM) and NOB are phylogenetically not closely related, and none Nitrospira exist, of which lineage II seems to be most widely distrib- of the environmental abundance and distribution of ammonia- in Nitrospira7. In both species all these genes were localized on a sin-
of these organisms can oxidize both substrates. This separation of the uted in both natural and engineered ecosystems10. The ecological suc- oxidizing microorganisms. Furthermore, the discovery of the long- gle contiguous genomic fragment, along with general housekeeping
two nitrification steps in different organisms leads to a tight cross- cess of Nitrospira has been linked to an economical pathway for nitrite sought-after comammox process will change our perception of the genes that allowed reliable phylogenetic assignment. Consequently,
feeding interaction and the frequently observed co-aggregation of oxidation17 and a substantial metabolic versatility, which includes nitrogen cycle. these Nitrospira species had the genetic potential for the complete
AOM with NOB in nitrifying consortia4. However, the functional the utilization of various organic compounds in addition to nitrite Nitrification, the aerobic oxidation of ammonium to nitrate is oxidation of ammonia to nitrate. No AMO of canonical ammonia-
separation is a puzzling phenomenon since complete nitrification and CO210,11,17–19, cyanate or urea degradation and nitrification by divided into two subsequent reactions: ammonium oxidation to oxidizing bacteria or archaea could be detected in the trimmed
would yield more energy (∆G°′ = −349 kJ mol−1 NH3) than either reciprocal feeding with AOM19,20, and chemolithoautotrophic aerobic nitrite (equation (1)) and nitrite oxidation to nitrate (equation (2)). metagenomic reads or by amoA-specific PCR8,9 on DNA extracted
single step (∆G°′ = −275 kJ mol−1 NH3 for ammonia oxidation hydrogen oxidation21. These two reactions are catalysed by physiologically distinct clades of from reactor biomass, and no other indications for the presence of
to nitrite and ∆G°′ = −74 kJ mol−1 NO2− for nitrite oxidation to microorganisms. ammonia-oxidizing microorganisms were found in the metagenome
nitrate). Thus, an organism catalysing complete nitrification should Enrichment of conspicuous Nitrospira or by FISH analyses. The AMO structural genes (amoCAB) of both
have growth advantages over the ‘incomplete’ AOM and NOB. Based A microbial biofilm developing on the walls of a pipe under the flow NH 4+ + 1.5O2 → NO2− + H2O + 2H+ (∆G°′ = − 274.7 kJ mol –1) (1) Nitrospira species, along with the putative additional AMO subunits
on kinetic theory of optimal pathway length5,6, Costa et al.7 argued of hot water (56 °C, pH 7.5) raised from a 1,200 m deep oil exploration amoEDD210,11, formed one gene cluster with haoAB-cycAB (encoding
that a hypothetical complete nitrifier would likely be outcompeted well (Aushiger, North Caucasus, Russia) was sampled and incubated NO2− + 0.5O2 → NO3− (∆G°′ = − 74.1 kJ mol−1) (2) HAO, the putative membrane anchor protein HaoB, electron trans-
by incomplete, cross-feeding AOM and NOB in many environments. at 46 °C in ammonium-containing mineral medium to enrich mod- NH 4 +
+ 2O2 → NO3− + H2O + 2H+ (∆G°′ = − 348.9 kJ mol ) –1
(3) fer protein cytochrome c554 and quinone-reducing cytochrome cm552,
However, the same authors7 also pointed out that a complete nitrifier erately thermophilic AOM. After a series of subcultivation steps, we respectively)12 and showed highest similarities to their counterparts
might be competitive under conditions that favour the maximiza- obtained enrichment culture ‘ENR4’ that oxidized ammonia to nitrate Even though the existence of a single microorganism capable of oxidiz- in betaproteobacterial ammonia-oxidizing bacteria (60% average
tion of growth yield rather than growth rate and coined the term and contained a dense population of cells morphologically resem- ing ammonium to nitrate (equation (3)) was not previously reported, amino acid identity to the Nitrosomonas europaea genes; Fig. 3 and
“comammox” (complete ammonia oxidizer) to describe such a bling described Nitrospira species13,15 (Extended Data Fig. 1a, b). it was proposed that such a microorganism could have a competitive Supplementary Table 1). The same genomic region also contained
hypothetical microbe. Conditions selecting for comammox may A second abundant population consisted of rod-shaped cells, but advantage in biofilms and other microbial aggregates with low substrate genes for copper and haem transport, cytochrome c biosynthesis, and
be characterized by slow, substrate-influx-limited growth with a no organism in ENR4 displayed the typical morphologies of known concentrations3. iron storage. These accessory genes were highly conserved in ammonia-
spatial clustering of biomass in microbial aggregates and biofilms7. AOM. Inspection by fluorescence in situ hybridization (FISH) with In this study, to characterize the microorganisms responsible for oxidizing bacteria but not in other Nitrospira7,13, indicating their
A prerequisite for the existence of comammox would also be that any nitrifier-specific ribosomal RNA-targeted probes10,22 confirmed that nitrogen transformations in an ammonium-oxidizing biofilm, we involvement in AMO and HAO biosynthesis or activation. Nitrospira
biochemical incompatibilities of ammonia and nitrite oxidation can ENR4 contained Nitrospira (Extended Data Fig. 1c) but no other sampled the anaerobic compartment of a trickling filter connected sp.1 encoded three discrete amoC genes, one of which was clustered
be overcome by adaptations of enzymes or cellular compartmentali- detectable nitrifiers. Moreover, known bacterial or archaeal genes to a recirculation aquaculture system4 with an ammonium effluent of with a second, almost identical copy of amoA (97.7% amino acid
zation7. Aside from these theoretical considerations, the old question of ammonia monooxygenase (AMO) subunit alpha (amoA) and 16S less than 100 µM. To enrich for the N-cycling community, a bioreactor identity). Nitrospira sp.2 lacked the second amoA, but contained four
of whether comammox exists in nature has not been resolved. rRNA genes of AOA were not detected by PCR in ENR4. Considering was inoculated and supplied with low concentrations of ammonium, additional amoC and a second haoA gene (Supplementary Table 1).
nitrite and nitrate under hypoxic conditions (≤3.1 µM O2). Within Unlike other Nitrospira7,13, both species lacked enzymes for assimila-
1
Department of Microbiology and Ecosystem Science, Division of Microbial Ecology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. 2Winogradsky Institute of Microbiology, Research 12 months, we obtained a stable enrichment culture that efficiently tory nitrite reduction, indicating adaptation to ammonium-containing
Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, bld. 2, 119071 Moscow, Russia. 3Center for Microbial Communities, Department of Chemistry and Bioscience,
Aalborg University, Fredrik Bajers Vej 7H, 9220 Aalborg, Denmark. 4Helmholtz-Centre for Environmental Research - UFZ, Department of Proteomics, Permoserstrasse 15, 04318 Leipzig, Germany. 1
Department of Microbiology, IWWR, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands. 2Center for Microbial Communities, Department of Chemistry and Bioscience,
5
Helmholtz-Centre for Environmental Research - UFZ, Department of Metabolomics, Permoserstrasse 15, 04318 Leipzig, Germany. 6Department of Microbiology and Ecosystem Science, Division
Aalborg University, Fredrik Bajers Vej 7H, 9220 Aalborg, Denmark. 3Laboratory for Microbiology, University of Gent, K. L. Ledeganckstraat 35, 9000 Gent, Belgium. 4Department of Biotechnology,
of Computational Systems Biology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. 7DVGW-Forschungsstelle TUHH, Hamburg University of Technology, Am Schwarzenberg-Campus 3,
TU Delft, Julianalaan 67, 2628 BC Delft, the Netherlands.
21073 Hamburg, Germany.
2 4 / 3 1 D E C E M B E R 2 0 1 5 | VO L 5 2 8 | NAT U R E | 5 5 5
5 0 4 | NAT U R E | VO L 5 2 8 | 2 4 / 3 1 D E C E M B E R 2 0 1 5
© 2015 Macmillan Publishers Limited. All rights reserved
© 2015 Macmillan Publishers Limited. All rights reserved
8

Energy yielding two-step oxidation process from ammonia to nitrate :

2 NH4+ + 3 O2 è 2 NO2- + 2 H2O + 4 H+ (nitritation) Nitrosomonas
Autotrophic bacteria Nitrobacter, Nitrospira
If summed up (mostly sufficient)
2 NH4+ + 4 O2 è 2 NO3- + 2 H2O + 4 H+ „Nitrifiers“
(4 ∙ 32) / (2 ∙ 14.01) = 4.57 g O2/g N oxidized

with
à 3.43 g O2/g N used for nitrite production and
1.14 g O2/ g NO2-N oxidized
9
Sequencing batch/MBR Process Overview

• Hybrid system coupling:
– Sequencing Batch Reactor (SBR)
– Membrane Bioreactor (MBR)
• Time managed, sequential process
• Provides flexibility to easily change
operational set points MBR
Membrane
Qi Qrecirculation Qpermeate
n
React Fill Stage React Draw Stage
SBR
Anaerobi Aerobic Anoxi Aerobi Anoxic Anoxi
c c c c
0 1 2
Cycle Time [hours]
Vuono et al. (2013). J. Membrane Science 446, 34

Impact of SRT on Microbial Diversity

• Amplicon sequencing of the
SSU rRNA (Phylogenetics)
• Disturbance followed by
recovery of SRT can restore
MLSS concentrations and
desired functions (COD- and N-
removal)
• Shortest SRT exhibits lowest

degree of diversity (Hill number)
• Same SRTs can result in

different degrees of diversity
while preserving general
function
(“functional redundancy
hypothesis” – Allison and Martiny,
Vuono et al. (2015), ISME 9, 425 2008)
Major Bacterial Phyla
Vuono et al. (2015), ISME 9, 425


Estimation of the amount of alkalinity required to carry out the reaction by rewriting the equation (neglecting
cell tissue):
NH4+ + 2 HCO3- + 2 O2 è NO3- + 2 CO2 + 3 H2O
For each g of ammonia nitrogen (as N) converted,
7.14 g of alkalinity as CaCO3 will be required (2 ∙ 50 g CaCO3 / 14)
13

Biomass synthesis reaction (using C5H7NO2 as an expression for bacterial cell synthesis):
4 CO2 + HCO3- + NH4+ + H2O è C5H7NO2 + 5 O2
Expression of an equation for the overall nitrification reaction (biomass synthesis and energy production):
NH4+ + 1.863 O2 + 0.098 CO2 è 0.0196 C5H7NO2 + 0.98 NO3- + 0.0941H2O + 1.98 H+
For each g of ammonia nitrogen (as N) converted,
à 1.863∙32 / 14 = 4.25 g of O2 are utilized and
0.0196 ∙ 113/14 = 0.16 g of new cells are formed
14

The oxygen required to oxidize 1.0 g of ammonia nitrogen (= 4.258 g) is
à less than the theoretical value of 4.57 g
à The alkalinity required for nitrification is 7.07 g/g instead of 7.14 g
The Yield coefficient (Ya = 0.23 g COD biomass/nitrate-N produced) of the nitrifiers expressed as the nitrifiers produced per
nitrogen oxidized to nitrate using the above equation:
Ya = (0.0196 ∙ 113 ∙ 1.42 ) / (0.98 *14.01) = 0.23 g/g
Nitrification causes a significant oxygen consumption (4.3 to 4.5 kg O2 / kg NH4-N, see above), which may vary strongly
during the day
The design of the aeration equipment has to take these aspects into account (adaption of the aeration rate along the
reactor length)
15

Matrix representation
With the Yield coefficient YA the growth process can be described by a kinetic matrix, such as:
Process Autotrophic O2 Nutrients Nitrate

Biomass [O2,-COD] [NH4-N] [NO3-N]
[COD]
Autotrophic 1 (YA-4.57)/YA -iN-1/YA 1/YA ρautotrophic

growth growth
Where
iN = nitrogen content of the COD fractions, 0.065 g N/g COD
YA = Yield coefficient for autotrophs
Biomass is expressed here in COD
Decay processes can be taken into account similar to heterotrophic decay
16
Nitrification – growth kinetics (suspended or attached)

The rate expression for growth ρautotr. Growth and decay ρend respiration (or ρdecay in general) may incorporate all limiting substances
(e.g., by a multiplication of Monod type terms and biomass, see previous lectures)
Rate expression as defined in the design procedure applied in this lecture (Metcalf & Eddy):
If dissolved oxygen is assumed to be present in excess and alkalinity is also not rate-limiting, the kinetic rate (growth and
decay) can be expressed as:
However, if dissolved oxygen is considered (nitrification rates increase up to DO concentrations of 3 to 4 mg/L):
17
Nitrification – some aspects

• Bacteria responsible for nitrification grow much more slowly than heterotrophic bacteria (μmax)
• Nitrification is pH sensitive and rates decline significantly at pH values below 6.8. At pH values near 5.8 to
6.0, the rates may be 10 to 20% of the rate at pH 7.0
• Nitrifiers tentatively inhibit themselves (if the buffer capacity of the water is not sufficient) because their
reactions set free protones, which reduces the pH of water
• Toxicity: Nitrifiying organisms are sensitive to a wide range of organic and inorganic compounds (at
concentrations well below the concentrations that would affect aerobic heterotrophic organisms)
• Systems designed for nitrification generally require much longer hydraulic and solids retention times
than those for systems designed only for BOD removal !!!
• Typical design SRT values for nitrifiying systems may range from 10 to 20 d at 10°C to 4 to 7 d at 20°C
18
Comparison of kinetic parameters for design purposes

Activated sludge kinetic coefficients
for heterotrophic bacteria at 20°C
Activated sludge nitrification kinetic

coefficients at 20°C
Source: Metcalf & Eddy (2000) 19

Comparison of growth rates

6.0
Wachstumsgeschwindigkeit [1/d]
autotrophe Bakterien
5.0 heterotrophe Bakterien
4.0
3.0
2.0
1.0
0.0
0 10 20 30 40 50 60 70 80 90 100
Substratkonzentration [mg/l]
20
Example for Concentration Profiles in a Simple Nitrifying System
O2
BOD5
NH4+
NO3-
(qualitative schematics! No CSTR, profiles simplified) 21

Nitrification: Required Environmental Conditions

Nitrification
aerobic
O2 ●
Nitrifiers ●
NH4+ ●
NO3- Product
HCO3- ●
Sludge age 7-10 d
Souce: Gujer, 1999 22

Biological Denitrification
23
Biological Denitrification – Process description

• Bacteria: heterotrophic bacteria in suspended and attached growth processes
• Denitrification is the biological reduction of nitrate to nitric oxide, nitrous oxide, and nitrogen gas:
NO3- à NO2- à NO à N2O à N2

• Heterotrophic growth of facultative aerobic organisms in the anoxic zones with nitrate as electron acceptor
instead of oxygen
• Denitrification therefore requires:
(1) At least two different environmental conditions (aerobic for nitrification and anoxic for denitrification)
(2) Nitrate gets into contact with organic degradable substances
24
Pre-anoxic Denitrification
Pre-anoxic denitrification (Modified Ludzak-Ettinger (MLE) Process)
Nitrate feed
Aerobic /
Anoxic nitrification
• Most common process used for biological nitrogen removal in municipal wastewater treatment
• Organic substrate in the influent wastewater provides the electron donor for oxidation reduction reactions using nitrate
• Nitrate concentration in the effluent ≈ nitrate concentration in the recirulation à no complete denitrification possible, but
the higher the recirculation, the better the denitrification
25
Gujer (2012) 26
7-10
DAFT
Underflow
Superna-
tant Return
Gravity
Thickener
27
Post-anoxic Denitrification
Post-anoxic denitrification
Aerobic /
nitrification Anoxic
• BOD removal has occured first and is not available to drive the nitrate reduction reaction
• Denitrification occurs after nitrification and the electron donor source is from (anoxic) endogenous decay (however,
uptake by this process is relatively slow)
• Often, an exogenous carbon source, such as methanol or acetate, is added to provide sufficient BOD for nitrate
reduction
28
Denitrification – Stoichiometry
Energy-yielding reaction (expressed here using e.g. C10H19O3N as an representation for biodegradable organic matter in
wastewater):

For wastewater:
C10H19O3N + 10 NO3- è 5 N2 + 10 CO2 + 3 H2O + NH3 + 10 OH-
Or for methanol:
5 CH3OH + 6 NO3- è 5 CO2 + 7 H2O + 3 N2 + 6 OH-
Or acetate:
5 CH3COOH + 8 NO3- è 10 CO2 + 6 H2O + 4 N2 + 8 OH-
29
C10H19O3N + 10 NO3- è 5 N2 + 10 CO2 + 3 H2O + NH3 + 10 OH-
1 equivalent of alkalinity is produced per equivalent of NO3-N reduced, which equates to 3.57 g of alkalinity (as CaCO3)
production per g of nitrate nitrogen reduced
In nitrification, 7.14 g alkalinity (as CaCO3) was consumed per g of NH4-N oxidized
à By denitrification about 50% of the amount transformed by nitrification can be recovered
30
Oxygen equivalent of using nitrate as electron acceptors
Metabolism of anoxic and aerobic growth of heterotrophs differs only in the energy yielding process. Per mole e- transferred
the following equivalents hold:
For oxygen:
0.25 O2 + H+ + e- è 0.5 H2O
For nitrate:
0.20 NO3- + 1.2H+ + e- è 0.1 N2 + 0.6 H2O
à 0.25 mole of O2 is equivalent to 0.2 mole of nitrate for electron transfer in oxidation reduction
Thus, the oxygen equivalent of nitrate is:

(0.25 ∙ 32 g O2 / mole)/(0.2 ∙ 14 g N / mole) ) = 2.86 g O2/g NO3-N
à Part of the oxygen that has to be put into the system can be saved because of denitrification
31
Oxygen equivalent of using nitrite as electron acceptors
For oxygen:
0.25 O2 + H+ + e- è 0.5 H2O
For nitrite:
0.33 NO2- + 1.33 H+ + e- è 0.67 H2O + 0.17
N2
The oxygen equivalent of nitrite is:
(0.25 ∙ 32 g O2 / mole)/(0.33 ∙ 14 g N / mole) ) = 1.71 g O2/g NO2-N
32
Estimating COD needed for Denitrification

The principal design goal in biological denitrification is the biological reduction of nitrate!
Thus, an important design parameter for denitrification processes is the amount of bsCOD or BOD needed to provide a
sufficient amount of electron donor for nitrate removal.
As a general rule, 4 g of BOD is needed per g of NO3 reduced!
The actual value will depend on the system operating conditions and the type of electron donor used for denitrification.
E.g., activated sludge process according to DWA A131:

Denitrification:
requirement for external carbon Þ 5 kg COD per kg of nitrate nitrogen to be denitrified
33
Estimating COD Needs for Denitrification

bsCODr = bsCODsyn + bsCODo
where
bsCODr = bsCOD utilized, g bsCOD/d

bsCODsyn = bsCOD incorporated into cell synthesis, g bsCOD/d
bsCODo = bsCOD oxidized, g bsCOD/d
bsCODsyn = 1.42 Yn bsCODr
Yn = net biomass yield, g VSS/g bsCODr (defined in previous lecture)

1.42 = conversion factor as introduced in previous lectures
Y
and Yn =
1 + (kdn) SRT
34
Thus,
bsCODr = bsCODo + 1.42 Yn bsCODr
and after rearrangement:
bsCODo = (1 - 1.42 Yn ) bsCODr
bsCODo is the COD oxidized and is equal to the oxygen equivalent of the NO3-N used for
bsCOD oxidation. Hence,
bsCODo = 2.86 NOx

with 2.86 being the O2 equivalent of NO3-N [g O2/g NO3-N] as given earlier
and NOx = NO3-N reduced [g/d]
à 2.86 NO3-N reduced = (1 - 1.42 Yn ) bsCODr

or 2.86
bsCOD 2.86
= and thus g bsCOD / g NO3-N =
NO3-N 1-1.42Yn 1-1.42Yn
35
Biological Denitrification – Stoichiometry

Matrix representation:
The process can be described by a kinetic matrix such as:
Process Substrate Hetero- O2 Nutrients NO3-N

[COD] trophic [O2,-COD] [NH4-N]
Biomass
[COD]
Heterotr. - 1/YH 1 - iN (-1/YH+1) / ρanox heterotr

anoxic growth 2.86 growth
(Denitrification)
Where iN = nitrogen content of the COD fractions, gN/gCOD
Biomass is expressed here in COD
Moreover, anoxic decay processes can be taken into account (see also postanoxic denitrific.) and are similar to aerobic
decay processes, but are typcially slower than aerobic processes.
36
Denitrification – Growth Kinetics

The rate expression for growth ρanox autor. Growth and decay ρend respiration (or ρdecay in general) may incorporate all limiting
substances (e.g., by a multiplication of Monod type terms and biomass, see previous lectures)
Rate expression as defined in the design procedure applied in this lecture (Metcalf & Eddy):
η = anoxic correction factor, gVSS/gVSS
η : 0.2 to 0.8 for pre-anoxic denitrification reactors fed domestic wastewater; takes
into account that
- only a portion of X is capable of dentrification (that fraction of denitrifiying bacteria
in the biomass), and that
- the maximum specific substrate utilization rate may be lower when nitrate is used
as an electron acceptor instead of oxygen
If the effect of nitrate and dissolved oxygen concentrations is to be incorporated into the rate expression :
Ko‘ = DO inhibition coefficient for nitrate reduction, mg/l

KS,NO3 = half velocity coefficient for nitrate limited reaction, mg/l
Other terms defined earlier
37
Example for Concentration Profiles in a Denitrifying System (preanoxic)

(qualitative schematics, no CSTR !)
O2
BOD5
NH4+
NO3-
Comment:
In this example denitrification is limited due to the supply of nitrate (concentration is zero whereas organic substances are still available).
This could be improved by an increase of the recirculation rate bringing more nitrate into the denitrification zone
Source: Gujer (2007) 38

Denitrification – some aspects

• Denitrification is especially fast, when large amounts of dissolved, readily biodegradable, organic
substances are available
• The volume for nitrification has to be approximatly the same as the nitrification volume without
denitrification
• N2 is only poorly soluble in water. As a result of denitrification gas bubbles can be generated and
massively disturb performance of biological treatment plants (especially when happening in the secondary
clarifier)
39
German Technical Design Guideline DWA A 131:
Index
:
40
Denitrification: Required Environmental Conditions

Denitrification
anoxic
O2 inhibits
COD ●
Het. Biomass ●
NO3- ●
HCO3- Product
Sludge age 10-20 d
Source: Gujer, 1999 41

ever, appreciable quantities are only released under acidic oxidizing (so-called Anammox) bacteria8 (planctomycetes)
Chair of Urban Water
23 Systems Engineering
conditions
Technical University ofand
Munichsuch conditions are uncommon in BNR or by DB.30 Frequently, these treatments are carried out
systems. simultaneously in single-stage systems.31,32 From a biochemical
Nitrogen Cycle
Fig. 1 Schematic representation of the proposed terminology. The continuously lined arrows indicate microbially catalyzed reaction pathways,
the dashed
Weißbach et al.lined arrows
(2017). Env. Sci.chemical
Wat. Res. &reactions,
Techn. 10, 3reaction pathways are indicted by dash-dotted lines. In yellow: The aerobic and sub-oxic reaction 42
Alternative Nitrogen Removal Processes:

Anaerobic Ammonium Oxidation (Anammox) or Deammonification
2 NH4+ + 4 O2 è 2 NO3- + 2 H2O + 4 H+ „Nitrifiers“
à (4 ∙ 32) / (2 ∙ 14.01) = 4.57 g O2/g N oxidized

with
3.43 g O2/g N used for nitrite production and
1.14 g O2/g NO2-N oxidized
2 NH4+ + 3 O2 è 2 NO2− + 2 H2O + 4 H+ „Shannon process“
NH4+ + NO2− è N2 + 2 H2O „Anammox

process“
à (3 ∙ 32) / (2 ∙ 14.01) = 3.43 g O2/g N oxidized
Tomorrowater.com
43
Biological Phosphorus Removal
44
Biological P Removal – General Process Description

• Chemical treatment to remove phosphorus is still the most commonly used technology
• Biological phosphorus removal has some advantages over the chemical removal (reduced chemical
costs and less sludge production as compared to chemical precipitation)
• Principle:
- Phosphorus in the influent wastewater is incorporated into cell biomass in high amounts
under certain conditions
- P is removed from the system with the waste sludge
• Bacteria: A number of bacteria is capable of storing polyphosphates. The P content of these bacteria
can amount to up to 15% („normal“ bacteria have a P content of 1-2%)
• These bacteria are grown in the treatment system and separated from the wastewater after they have
stored large amounts of P
45
Biological P Removal – General Process Description

A simplified way to describe the process is:
Properties of the P-assimilating organisms ( PAOs)
(1) Capable of storing excess amounts of P as polyphosphates in their cells.
(2) Under anaerobic conditions, PAOs assimilate fermentation products (e.g., volatile fatty acids)
into storage products within the cells with the concomitant release of phosphorus from stored
polyphosphates
(3) Under aerobic conditions, PAOs produce energy by the oxidation of storage products and
polyphosphate storage within the cell
46
Biological Phosphorus Removal Technology

Simplified biochemical model:
ANOXIC
ANAEROBIC PROCESS AND AEROBIC PROCESS
dissolved ortho- NO3- ortho-

substrate phosphate O2 phosphate
energy cell growth
energy
Poly- Poly- Poly- Poly-
substrate phosphate substrate phosphate


• The phosphate accumulating organisms (PAOs) have the possible advantage (different from the
heterotrophic organisms) that they can store substances under anaerobic environmental conditions and
assimilate acetate while other organisms are starved (therefore, a portion of the bsCOD is taken up
by the PAOs)
• These substances then allow them to grow under aerobic conditions
The process must be designed in a way that this property can be used by the organisms as an
advantage in order to grow them in the system
48
Biological P Removal – Process Description
effluent
dissolved ortho-
substrate phosphate ortho-
O2 phosphate
energy cell growth
energy
Poly- Poly- Poly- Poly-

substrate phosphate substr phosphate
ate
Source: Metcalf & Eddy (2000) and Gujer (2007) 49


NO3
COD
bio-P DN nitrification
Periodic exposure of the activated sludge organisms to selected process conditions allows control of
structure and function of heterogeneously composed microbial communities.
Note:
The preferred food source of the PAOs (low molecular weight fermentation products) would not be available
without the anaerobic zone!
The anaerobic zone provides for the fermentation of the influent bsCOD to acetate.
50
Example for concentration profiles in a system with biological P

elimination (UCT process)
XTSS
NO33--
NO
PO
PO44
bsCOD
(qualitative schematics)
Souce: Gujer (2007) 51
Source: Yang Jiang (2011) Polyhydroxyalkanoates production by bacterial enrichments (PhD Thesis TU Delft) 52
Fate of soluble BOD and phosphorus in reactor
53
Biological P Removal – Stoichiometry

• The amount of phosphorus removed by biological storage can be estimated in a simplified way from the
amount of bsCOD that is available in the wastewater influent (most of the bsCOD will be converted to
acetate in the anaerobic reactor)
• With the following assumptions:

(1) 1.06 g acetate/g bsCOD will be produced (most of the COD fermented will be converted to VFAs due to
the low cell yield of the fermentation process).
(2) A cell yield of 0.3 g VSS/g acetate
(3) A cell phosphorus content of 0.3 gP/g VSS
• Using these assumptions, about 10 g of bsCOD will be required to remove 1 g of phosphorus by the
mechanism of biological storage
• Normal cell synthesis by uptake of bCOD in the activated sludge system will result in additional
phosphorus removal
54
Biological P Removal – Growth kinetics

• Biological phosphorus growth kinetics are within the same order of magnitude of other heterotrophic
bacteria
Conditions:
1) Dissolved organic biologically degradable substances (volatile acids),

No Nitrate and No Oxygen must be available !
2) A subsequent reactor (aerobic or anoxic) with enough reaction time
3) The biomass must be adapted to the process (weeks)
4) The phosphorus rich sludge must be taken out of the system under aerobic conditions
For further reading on (dynamic) growth kinetics (simulation) models for biological P removal see:
-ASM 2d (IWA Task Group)

-Bio P-Model of TU Delft and related publications
55
Biol. P Removal: Required Environmental Conditions

Biological P Elimination
phase 1 phase 2
anaerobic aerobic
O2 inhibits ●
COD ●
PAO ● ●
NO3- inhibits
PO43- Product ●
HCO3- ●
Sludge age > 15 d
EXAMPLE
Estimating the amount of phosphorus removal
Given the following influent wastewater characteristics and the corresponding biological
process information, estimate the effluent phosphorus concentration.
•Heterotrophic synthesis yield, Y = 0.40 g VSS/g COD
•Endogenous decay coefficient, kd = 0.08 g VSS/g VSS.d
•SRT = 5 d
•Phosphorus content of PAOs = 0.30 g P/g VSS
•Phosphorus content of other bacteria = 0.02 g P/g VSS
•Clarifier effluent VSS concentration = 8 g/m3
Influent Concentration, g/m3

COD 300
bCOD 200
bsCOD 50
Phosphorus 6
57
Estimate the effluent phosphorus concentration. Heterotrophic synthesis yield, Y = 0.40 g VSS/g COD;
Endogenous decay coefficient, kd = 0.08 g VSS/g VSS.d; SRT = 5 d; P content of PAOs = 0.30 g P/g VSS;
P content of other bacteria = 0.02 g P/g VSS; Clarifier effluent VSS concentration = 8 g/m3
Solution
Y
Yn =
1 + (kdn) SRT
Yn = net biomass yield, g VSS/g bsCOD
58
59
60
Required Environmental Conditions

Aerobic Nitrification Denitrificatio Biological P
biological n Elimination
oxidation phase 1 phase 2
aerobic aerobic anoxic anaerobi aerobic
c
O2 ● ● inhibits inhibits ●
COD ● ● ●
Het. Biomass ● ●
Nitrifiers ●
PAO ● ●
NH4+ ●
NO3- Product ● inhibits
PO43- Product ●
HCO3- ● Product ●
Source: Gujer, 1999 Sludge age >5d 7-10 d 10-20 d > 15 d 61
Required System Configurations

Aerobic Nitrification Denitrification Bio P removal
biological
oxidation
Aerobic
conditions ● ● ● ●
Anoxic
conditions ● ●*
Anaerobic
conditions ●
Sludge age >5d 7-10 d 12-18 d
*Biological phosphorus removal is not used in systems that are designed with nitrification without including a means for
denitrification to minimize the amount of nitrate in the return sludge flow to the anaerobic zone.
62
Dynamic Activated Sludge Models

1 February 2024
1
Steps to Build a Dynamic WWTP Model

1. Definition of the WWTP model purpose or the objectives of the model application (design, reconstruction,
control, learning)
2. Model selection, which means the selection of appropriate models needed to describe the different
WWTP units that should be considered in the simulation, i.e., selection of the activated sludge model, the
sedimentation model, etc.
3. Hydraulic determination of the models for the whole plant or specific units of WWTP (i.e., hydraulics of
the aeration tanks, secondary settling tanks, etc.)
4. Determination of the wastewater and biomass characterization, including biomass sedimentation (influent
and effluent data, design parameters, etc.)
5. Calibration and verification of the activated sludge model parameters
6. Evaluation of the different WWTP scenarios (i.e., optimization of the WWTP operation, process
improvement, different technical (structure) solution, etc.)
Source: A. Serdarevic & A. Dzubura (2016). Wastewater Process Modeling. Coupled Systems Mechanics 5(1), 21-39 2
IWA Activated Sludge Models ASM1, ASM2, ASM3

≥
ftp://ceres.udc.es/Master_en_Ingenieria_del_Agua/master%20antiguo_antes%20del%202012/Segundo_Curso/Programas_Comerciales_en_Ingenieria_Hidraulica_y_Sanitaria/ajacome-aquasim/MODELOS-ASM1-ASM2-ASM3.pdf
3
SIMBA# Version 3.0 Process Simulator

for Modeling, Simulation and Optimization
Selected Features:
• Detailed fully dynamic aeration system model library (incl. blowers, pipes,
valves, etc.) Introduction to SIMBA:
• Reactor models for CSTR, MABR, MBBR, MBR, AGSR, trickling filter, etc.
https://www.youtube.com/watch?v=ELjlix04nE8
• Library for process water management and treatment
• Advanced Gujer process matrix editor https://www.youtube.com/watch?v=93MGzC2hxf0
• Tools for result evaluation and LCA studies
• Pollutants in sewers and rivers
Source: www.ifak.eu; inCTRL Solutions, Inc. 4
SUMO Wastewater Process Simulator
Selected Features:
• Most extensive calibrated model library for traditional and advanced wastewater resource recovery processes
• Sumo is the only open process source commercial simulator2 (coded in Excel tabular format in SumoSlangTM)
• Virtually unlimited activated sludge/anaerobic digester/sidestream treatment configurations available
• Integrated steady-state and dynamic simulations, book of SumoSlang, extended manual, software in Korean, Chinese,
Japanese, Spanish, Turkish
Source: http://www.dynamita.com/the-sumo/ 5
WRc Ltd STOAT

Wastewater Process Simulator
Download request (Freeware):
http://sites.wrcplc.co.uk/freeware/STOAT/downloadform.aspx
Source: http://www.wrcplc.co.uk/ps-stoat 6
Example Manual reference:
BIOWIN Wastewater Process Simulator

Model Reference > Chemical Precipitation Reactions > Chemical
Phosphorus Removal with Iron Salts
Introduction to BioWin:
https://www.youtube.com/watch?v=cJ0dGY9w2Gg
Watch a BioWin application for a plant upgrade:
https://www.youtube.com/watch?v=0VgTvumjlpk
Source: https://envirosim.com 7
Hydromantis GPS-X Version 6.0
Source: https://www.hydromantis.com/GPSX.html 8
Hydromantis GPS-X Lite (open access)
https://www.hydromantis.com/GPS-X-Limited/download.html
Request form for free download (PC only):

https://www.hydromantis.com/GPS-X-Limited/request-form.html
9
Energy Recovery and Sludge Handling

8 February 2024
1
Solids Handling – Resource Recovery
2
Sludge Digesters at the Wastewater Treatment Plant

Munich II, Gut Großlappen
3
Environmental Conditions/Terminology
Energy source Conditions in the environment

Available electron acceptor being reduced
Respiration
a) O2 Aerobic
b) Nitrate, NO3-; nitrite, NO2-; no oxygen Anoxic
c) Sulfate, SO42-; CO2; no oxygen; no nitrate Anaerobic
Fermentation
d) Organic compounds, no oxygen, no nitrate Anaerobic
4
Anaerobic Digestion
General Process Description
• Microorganisms: facultative and obligate anaerobic bacteria (hydrolysis and fermentation), strict
obligate anaerobic (methane production)
• Primarily used for the treatment of waste sludge and high strength organic wastes
• Characterized by
lower biomass yields (less sludge production), and possibly
energy recovery through methane production (instead of energy used for aeration)
• often used as a pre-treatment step
• mostly operated in the mesophilic temperature range (30 to 40°C)
• Thermophilic fermentation (>50°C) also possible resulting in high pathogen inactivation
5
Anaerobic Digestion
Hydrolysis:
• Conversion of particulate matter to soluble compounds
Fermentation (Acidogenesis):
• Further degradation
• Organic substrates serve as both electron donor and acceptor
• Final products of fermentation are acetate, hydrogen, and CO2
Methanogenesis:
• Carried out by methanogens Harvestpower.com
• Two groups of methanogens:

1st: acetate à CH4 and CO2;
2nd: Hydrogen (electron donor) and CO2 (electron acceptor) à methane
6
Anaerobic Digestion
Complex Organics
(Carbohydrates, Proteins, Lipids)
Fermentative bacteria
(Hydrolysis)
Monosaccharides, Higher Fatty Acids,

Amino Acids Alcohols
Fermentative bacteria
(Acidogenesis)
Volatile Acids Acetogenic bact.
(Propionic, Butyric, etc.) (Acetogenesis)
Hydrogen-Producing Acetogens
H2 + CO2 Acetate
Hydrogen-utilizing acetogens
Methanogenic organisms (Methanogenesis)
Hydrogen utilizing CH4 + CO2

Aceticlastic
methanogens
methanogens
Von Sperling (2005)

Anaerobic Digestion
Process Description
Step 1 Hydrolysis
Step 2 Acidification
Step 3 Acetogenesis
Step 4 Methanogenesis
4 microorganism groups are needed !
4 microorganism groups have to build a

symbiotic community
Anaerobic Digestion
Anaerobic systems are characterized by complex interactions
The symbiotic co-existence requires certain conditions (important process parameters:
• Buffer capacity
• pH
• Propionic acid
• Hydrogen
• Acetate
• Redox potential
à Check feed conditions

à Mix, but without introducing extreme local shear forces
Anaerobic Digestion
Temperature
• the higher, the faster are the reactions
• as long as enzymes are not harmed Þ Optimal Temperature
• different bacteria types have different optima
Reaction-
rate
psychrophilic
Bacteria
opt.: ca. 16 °C extremophilic
thermophilic Þ Bacteria
Bacteria opt.: über 100 °C
opt.: ca. 57 °C
mesophilic
Bacteria
opt.: ca. 38 °C
Temperature, °C
20 40 60
Anaerobic Digestion
Anaerobic Fermentation/Oxidation – Stoichiometry
• A COD balance can be used to account for the changes in COD during fermentation
• Instead of oxygen, the COD loss in the anaerobic reactor is accounted for by the methane production
• The COD equivalent of methane is:
CH4 + 2 O2 à CO2 + 2 H2O
COD per mole of methane : 2 (32 g O2/mole) = 64 g O2/mole
Volume of methane per mole at standard conditions (0°C and 1 atm) = 22.414 L
à CH4 equivalent of COD : 22.414 / 64 = 0.35 L CH4 / g COD
• Synthesis yield coefficients for anaerobic processes are considerably lower compared to aerobic
processes (see also former lectures)
11
Anaerobic Digestion
Aerobic vs anaerobic treatment – carbon balance
CO2 Biogas
(40 - (70 -
COD 50%) COD 90%)
(100%) (100%)
Aerobic Anaerobic
Reactor Reactor
effluent effluent
(5 - 10%) (10 - 30%)
sludge sludge
(50 - 60%) (5 - 15%)
Von Sperling
Anaerobic Digestion
COD balance throughout the anaerobic degradation process
COD cells COD cells

COD
bCOD removed
Total COD VFA COD CH4
COD
COD VFA COD not

removed
COD rec COD rec COD rec
bCOD = biodegradable COD

CODrec = non biologically degradable COD and COD in complex substrate form treated
in anaerobic reactors containing biomass not yet adapted to the complex substrate
COD VFA = Fraction of COD still present as volatile fatty acids
CODCH4 = COD converted into methane
CODcells = COD converted into cells
Von Sperling (2005)

Anaerobic Digestion
Kinetic and stoichiometric parameters
µmax YS
d-1 kg VSS / kg COD
Aerobic bacteria
readily degradable substrate 10 - 20 0.4 – 0.6
Hydrolysers 0.1 – 1.5 0.02 – 0.2
Acidifying organisms 2-4 0.1 – 0.3
Methanogens 0.3 – 0.4 0.03 – 0.04
The weak organisms in the system!

Methanogenic growth kinetics are of main interest in the design of the reactor
Hydraulic detention time of approximately 20 – 30 d (>> 1 / µmax )

Anaerobic Digestion
Anaerobic Fermentation/Oxidation – Growth kinetics
• Two rate limiting concepts are important:
- Hydrolysis conversion rate

(can be decisive if compounds are difficult to hydrolyse, affects the total amount of
solids converted)
- Soluble substrate utilization for fermentation and methanogenesis

(affects stability of the process)
à Depending on the conditions (inflow composition), a two-step process can be advantageous to create
optimal conditions for the different stages in the process
15
Anaerobic Digestion
Pros and cons of anaerobic treatment
Advantage Disadvantage
Low production of solids Anaerobic bacteria are susceptible to
inhibition by many compounds
Low energy consumption Process start up can be slow
Low land requirements Post-treatment usually necessary
Low construction costs Complex biochemistry and microbiology
Methane production (energy) Possible generation of odours
Tolerance to high organic loads Possible generation of effluents with
unpleasant aspects
Small and large scale application Unsatisfactory removal of nitrogen and
phosphorus
Low nutrient consumption
Combination of aerobic and anaerobic processes to exploit advantages of both processes

(e.g. sludge digestion)
16
EXAMPLE
Prediction of methane gas production
An anaerobic reactor, operated at 35°C, processes a wastewater stream with a flow of
3,000 m3/d and bsCOD concentration of 5,000 g/m3. At 95 percent bsCOD removal and a
net biomass synthesis yield of 0.04 g VSS/g COD used, what is the amount of methane
produced in m3/d?
17
An anaerobic reactor, operated at 35°C, processes a wastewater stream with a flow of 3,000
m3/d and bsCOD concentration of 5,000 g/m3. At 95 percent bsCOD removal and a net biomass
synthesis yield of 0.04 g VSS/g COD used, what is the amount of methane produced in m3/d?
Solution
18
An anaerobic reactor, operated at 35°C, processes a wastewater stream with a flow of 3,000
m3/d and bsCOD concentration of 5,000 g/m3. At 95 percent bsCOD removal and a net biomass
synthesis yield of 0.04 g VSS/g COD used, what is the amount of methane produced in m3/d?
19
Anaerobic Digestion
• Anaerobic Processes are cost effective, but only if the influent concentration is relatively
high (> 2,000 mg COD/L)
• At 30°C and at normal pressure (1.0133 bar), approximately 20 g CH4 / m³ are soluble in water.
This corresponds to a COD of approxiately 80 g/m³
à Biogas is only generated, if at least 80 g/m³ of COD are degraded. Wastewaters with low
concentrations will therefore only produce little biogas!
20
Gas Collecting Systems

Anaerobic Digestion
Digester shapes
Cylindric shape
with flat bottom
Problem: This causes

Sediment accumulation in corners à Vusable < VReactor
qh real < qh required
Anaerobic Digestion
Digester shapes
Cylinder shape with

cones
Problem:
Complex reinforcement in the corners
Advantage:
• good mixing
• insensitive to settling
Anaerobic Digestion
Digester shapes
Egg-shape
Advantage:
• good mixing
• insensitive to settling
• low radiation losses
• stable structure
WWTP Köhlbrandhöft, Hamburg Harbour
25
Disposal Strategies of Digested Sludge in Bavaria
DWA (2017). Mitgliedermitteilungen 26

P-Recovery Strategies from Sludge
DWA (2017). Novellierung der AbfKlärV 27

Disinfection

8 February 2024

1
Disinfection
Drinking Water Treatment
Wastewater Treatment
2
History Recap…
• Koch (Germany, 1881) demonstrated the role of bacteria in waterborne diseases and
demonstrated that chlorine can inactivate pathogenic bacteria
• Filtration for control of waterborne diseases: low efficiency, even when combined with
coagulation
• 1902 – first continuous chlorination for disinfection of drinking water (Middelkerke,
Belgium)
• 1908 – first continuous chlorination in the US in Jersey City, NJ (solid calcium
hypochlorite, Ca(ClO)2)
• Liquefied chlorine gas makes large-scale operation more feasible (first facility in
Philadelphia, 1913)
• By 1941, 85% of drinking water supplies in the US were chlorinated
3
History Recap…
• In Germany, almost all water supplies come from protected sources (groundwater,
springs, riverbank filtration) favoring natural filtration and inactivation of pathogens
(“chemical disinfection” is option of last resort)
• Discovery in the US and Europe that DBP are formed when chlorine reacts with NOM
(mid 1970s)
• Limits are set by the EU Drinking Water Directive and the USEPA on THM
concentrations in drinking water
• More DBP are discovered and more regulations follow
• Last two decades of 20th century: questioning the effectiveness of chlorination in
controlling waterborne diseases caused by protozoa
4
Disinfection
• Two basic strategies to reduce microbiological contaminants in drinking water:
– removal from the water
– inactivation
• Inactivation ≡ traditionally referred to as disinfection

• Inactivation – still present but harmless to consumer
5
Disinfection
• Primary Disinfection: inactivation of microorganisms in the water
• Secondary Disinfection: maintaining a disinfectant residual in the
distribution system
• Five disinfection agents are commonly used in treatment of drinking
water:
– Free chlorine – most common
– Combined chlorine (not allowed in Germany)
– Ozone – strongest oxidant, controls T&O
– Chlorine dioxide
– UV irradiation
6
EU Regulations/German Trinkwasser Verordnung

• EU Drinking Water Directive/German TrinkwV
– Escherichia coli 0/100 mL
– Enterococci 0/100 mL
– Coliform bacteria 0/100 mL
– Clostridium perfringens 0/100 mL (only for surface water supply)
• German TrinkwV only

– Heterotrophic plate count (22/36°C) 100 HPC/mL
– Legionella spec. 100 units/100 mL (triggers additional measures)
– Somatic coliphage 50 PFU/100 mL
• Disinfection by-products (EU/German TrinkwV)

– Trihalomethane (THM) – 0.05 mg/L (up to 0.1 mg/L)
– Haloacetic acids (HAA-5) – 0.06 mg/L 7
U.S. Regulations
• Surface Water Treatment Rule
– treatment requirements:
• 3-log inactivation of Giardia lamblia cysts
• 4-log inactivation of enteric viruses
• Interim Enhanced Surface Water Treatment Rule

– treatment requirements:
• 2-log inactivation of Cryptosporidium
• Long Term 2 Enhanced Surface Water Treatment Rule

– treatment requirements for Cryptosporidium:
• Bin 1: <0.075 oocysts/L no additional treatment
• Bin 2: 0.075 to <1.0 1.0 log additional
• Bin 3: 1.0 to <3.0 2.0 log additional
• Bin 4: >3.0 2.5 log additional
8
Long Term 2 Enhanced Surface Water Treatment

Rule (LT2ESWR) – “Toolbox”
• Watershed Control Program (0.5-log)
• Pre-sedimentation basin with coagulation (0.5-log)
• Lime-soda softening (0.5-log)
• Bank Filtration (0.5-log to 1.0-log depending on set-back)
• Combined filter effluent turbidity less than or equal to 0.15 NTU in at least 95 percent of
measurements each month (0.5-log)
• Slow sand filters (2.5-log)
• Membranes
• Chlorine Dioxide (based on CT table)
• Ozone (based on CT table)
• UV (based on UV dose table)
9
U.S. Regulations
• Stage 1 Disinfectants and Disinfection Byproducts Rule (Stage 1 D/DBPR)
– Regulates disinfectants: Maximum Residual Disinfectant Level (MRDL)
– Regulates disinfection by-products (DBPs)
• TTHMs (0.08 mg/L)
• HAA5 (0.06 mg/L)
• Chlorite (1.0 mg/L)
• Bromate (0.01 mg/L)
10
Emerging Issues
• Chlorination/disinfection byproducts
• Chlorine-resistant organisms
• Both causing reevaluation of chlorine as a primary disinfectant and
reevaluation of the role of inactivation itself in the control of pathogens
• New methods for removal: membrane filtration, UV irradiation
11
Chlorine Reactions with Organic Constituents

(AOX - adsorbable organically bound halogens)
Behavior of AOX-subgroups during chlorination
0.59
160
15.8
140
0.17
120 AOI
11.27 AOBr
100 AOCl
AOX (ug/L)
80
134
60
101.2
40
0.78
20 4.79
12.3
0
Val Vista prior Cl2 Val Vista after Cl2 Val Vista finished water
12
Characteristics of Five Most Common Disinfectants
13
Characteristics of Disinfectants
• T13-1
14
Oxidation Power
Disinfectant Oxidation potential (Volt)
Hydroxyl radical -2.8
Ozone -2.07
Hydrobromous acid -1.59
Hydrochlorous acid -1.49
Chlorine -1.36
Bromine -1.07
Chlorine dioxide -0.95
Monochloramine -0.75
Dichloramine -0.74
15
Kinetics of Disinfection
• The specific mechanism of microorganism inactivation during chemical
disinfection is not well understood
• Function of chemical disinfection agent, properties of the
microorganism, and properties of the water
• One simple kinetic model is widely used, but many other models
describe the mechanism of disinfection
– Dr. Harriet Chick - Chick’s Law (1908) – disinfection could be modeled
as a pseudo-first-order reaction with respect to the concentration of
the organisms
16
Kinetics of Disinfection – Chick’s Law
dN
= -KN
dt
17
Kinetics of Disinfection – Chick’s Law

dN
= -KN
dt
dN/dt = rate of change in # of organisms with time
N = concentration of microorganisms, organisms/volume
K = Chick’s law rate constant, time-1
After integration:
æN ö
lnçç ÷÷ = -Kt
è N0 ø
18
19
Kinetics of Disinfection – Chick-Watson Law

C n t = constant
n ⋅ log(C) + log(t) = log(constant)
dN
= −k 'C n N
dt
C = concentration of disinfectant, mg/l
n = empirical constant, (-)
t = time required to achieve a constant % of inactivation
constant = value of given % of inactivation
After integration:
!N$ n N −k 'C nt
ln # & = −k 'C t _ or _ =e
" N0 % N0
20
Chlorine Disinfectants
• Chlorine
– Most widely used
– Effective at low concentrations
– Inexpensive
– Forms residual if applied in sufficient dosage
– Powerful oxidation properties
– Can be applied as a gas (Cl2(g)) or liquid (hypochlorite (bleach))
21
• Chlorine gas
– Liquified at 5-10 bar
– Shipped in steel cylinders (99.8% Cl2) containing 45, 68, 910 kg
Cl2(g) ó Cl2(aq) KH = Cl2(aq)/pCl2
Cl2 + H2O à HOCl +HCl

Hydrochloric acid
Hypochlorous acid
HOCl Û H+ +OCl¯
22
• Chlorine gas
– The relative distribution of HOCl and OCl¯ is pH
dependent
23
• Hypochlorite salts:
– Calcium hypochlorite – dry
– Sodium hypochlorite – liquid
Ca(OCl)2 à Ca2+ +2 OCl ¯
NaOCl à Na+ + OCl ¯

NaOCl + H2O à HOCl + Na+ + OH-
24
• Combined chlorine:
– Ammonia reacted with chlorine (max. 6 mg/L according to TrinkwV):
NH3 + HOCl à NH2Cl + H2O monochloramine

NH2Cl + 2 HOCl à NHCl2 + 2 H2O dichloramine
NHCl2 + 3 HOCl à NCl3 + 3 H2O nitrogen trichloramine
25
• Chlorine dioxide (ClO2):
– Three general methods of formation:
– NaClO2 + HCl à ClO2 + NaCl + H+
– (Cl2 +H2O à HOCl +HCl)

HOCl + HCl + 2 NaClO2 à 2 ClO2 + 2 NaCl + H2O
– 2 NaClO2 + NaOCl + 2 HCl à 2 ClO2 + 3 NaCl + H2
26
Chlorine Reaction
• As chlorine is added it reacts with constituents until residuals
are produced (Cl2, HOCl, OCl ¯ ≡ free chlorine)
• Residuals composed of chloramines ≡ combined available

chlorine residuals
• Sum of the above ≡ total residual chlorine (TRC)
27
Breakpoint Chlorination
28
Breakpoint Chlorination
• From point A to B: chlorine reacts with organic compounds, ammonia, and
amines to form chloro-organic species and chloramines. Free Chlorine is NOT
formed.
• From point B to minimum: dichloramine is decomposed to nitrogen
trichloramine and chloramines may also decompose to N2 and N2O
• Breakpoint: the point at which all destructible chloramines and chloro-organic
compounds are decomposed and FREE chlorine residual begins to appear
• Beyond breakpoint: free and combined residual in solution
29
Chlorine Demand
30
Classwork
xxxxxx
31
Classwork
The chlorine demand curve on the graph was obtained for a drinking water for a 1-hour contact
time. Determine the daily amount of NaOCl to be applied to this water in order to produce a free
and combined residual of 0.9 mg/L after a contact time of one hour in a flow of 24,000 m3/day.
1
0.9
0.8
Chlorine Residual, mg/L

0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0 0.5 1 1.5 2
Chlorine Dose, mg/L
32
CT Approach
33
Residual CT is calculated with a very conservative approach !

4
3
Cl2 (mg/L)
0
0 5 10 15 20
Time (minutes)
34
Integral CT
Cl2 (mg/L) 3
CT = 70 (mg-min/l)
0
0 5 10 15 20
Time (minutes)
35
Residual CT
3
Cl2 (mg/L)
1
CT = 50 (mg-min/l)
0
0 5 10 15 20
Time (minutes)
36
Residual CT
3
Cl2 (mg/L)
0
0 5 10 15 20
Time (minutes)
37
CT Approach
• “CT Value” used in water treatment – Conservative Estimate
• It is the product of the residual disinfectant concentration C (mg/L) determined at
the end of the process (reactor effluent), with the corresponding disinfectant contact
time T (minutes)
• Appropriate CT value must be satisfied at peak flow
• Best disinfection achieved in plug-flow reactor
• C = 0.2 – 40 mg/L, rarely exceeds 15 mg/L
• T = measure of the time at which 10% of a tracer feed concentration (T10) appears at
the sampling point
• CT has long been used as a basis for disinfection requirements
• Useful way of comparing alternative disinfectants
38
CT Approach
Removal of 99%
C*t = important design criteria! 39

CT Approach
log(C) = -log(t) +log(constant)
40
CT Approach
41
TTab for Inactivation of Viruses by Free Chlorine
42
TTab
43
TTab for 3-log Inactivation of Giardia Cysts by Free Chlorine
44
TTab
45
CT Values for Inactivation of Giardia Lamblia

Cysts and Viruses by
Chlorine, Dioxide, Chloramine, and UV
46
CT Approach
47
48
Ultraviolet Light (UV) Disinfection

• UV is effective against Cryptosporidium
• UV does not contribute to formation of
regulated DBPs
• Established technology
• Emerging UV-LED
• State of rapid development
49
UV Light in the Electromagnetic Spectrum

Gamma
Rays X - Ray UV Visible Infrared
254
Vacuum UV-C UV-B UV-A

UV absorbance
DNA
Water
100 200 300 400

Wavelength (nm)
50
Why Mercury Lamps?
• Electron transitions provide energy in the germicidal wavelength

range
• Low vapor pressure is needed for high efficient UV generation
• High vapor pressure can produce high intensity polychromatic light,

but lower efficiency
• Low ionization energy, easily start gas discharge at low voltage
• Hg is stable in the lamp envelope

51
Spectra of LP and MP Mercury Lamps
Relative Intensity (mW )cm

200 250 300 350 400
Low Pressure
Wavelength (nm)Hg Medium Pressure Hg
) -2
-2
Relative Intensity (mW )cm

Relative Intensity (mW cm
200 225 250 275 300 200 250 300 350 400
Wavelength (nm) Wavelength (nm)
monochromatic vs. polychromatic

-2
52
Low Pressure, High Intensity

• Low Pressure - Higher Wattage Lamps
• 0.13 to 0.79 kW/1,000 m3
• Lamp Life Expectancy greater than 5,000 -
12,000 hours
• Available in vertical, horizontal, and
combined alignment
• Self Cleaning System
53
54
Mechanism of UV Irradiance in Water
UV lamp
Absorbing
water column Quartz
sleeve
(fouling)
DNA target UV energy
Target
organism Particulates
Enlarged
target
organism
55
Calculation of UV Dose
D = Iavg t
Where D = UV dose (fluence), mJ/cm2 (note mJ/cm2 = mW s/cm2)
Iavg = average UV intensity, mW/cm2
t = exposure time, s
"I%
€ log$ ' = −ε( λ )Cx
# Io &
Where I = UV light intensity at distance x from the light source, mW/cm2

Io = UV light intensity at light source, mW/cm2
e(l) – molar absorptivity
€ of the light absorbing solute
C = concentration of light-absorbing solute, mole/L
x = pathway distance, cm
56
Inactivation
• First-order reaction:
Nt
ln = -kIt
No
– Nt and No are the number of the viable microorganism at time t and initially, respectively
– k is the inactivation constant
– I is the average UV intensity (irradiance) in the reactor
– t is the average detention time in the irradiated portion of the reactor
57
Water Quality Effects
Particulate Matter
Dissolved Matter
• iron
• nitrate
• natural organic matter
58
Short-Circuiting
59
Characteristics of Three Types of UV Lamps
60
UV disinfection systems
61
Lamps in Modules
62
Modules in a Bank
63
Flow Control in Serpentine

Contactor
64
Typical UV Doses Required to Achieve Different Coliform

Disinfection Levels for Different Wastewaters
65
Cr
yp UV Dose (mJ/cm2)
to
sp
or
50
0
100
150
200
250
id
iu
E. m
co Gia
li rd
Sa Sh O1 ia
lm ig 57
Le on ell :H
gi el a 7
on la so
el en nn
la t ei
pn eri
eu tid
m is
op
P H hi
ol ep la
io at
Co vir it
u is
xs s A
Typical UV Dose - 40 mJ/cm2
ac Ty
ki pe
R e 1
ot vir
A a us
de v B
UV Doses Required
no iru 5
s
UV Doses for 99.99% removal
vi SA
ru
s 11
ty
pe
41
66
Comparison to Other Disinfectants

1000
Bacteria
Viruses
Protozoans
Relative to E. coli
Dose Required -
100
Spores
10
0.1
Chlorine Ozone UV Light
Disinfectant Type
67
Summary
• How does UV compare to other current technologies?
68
BGU38010
Advanced Water Treatment Engineering and
Reuse
Summer term 2024, Thursdays, 9.45 – 11.15 h

(jdrewes@tum.de)
69
BV180004
Modeling of Aquatic Systems
SS 2024
Prof. Dr.-Ing. Konrad Koch

(k.koch@tum.de)
70
BGUxxx
Anaerobic Technology and Energy Recovery
Summer term 2024, Thursday afternoon
Prof. Dr.-Ing. habil. Konrad Koch

(k.koch@tum.de)
71
BGU xxx
Engineered Natural Treatment Systems
Winter Term 2024/25
Dr.-Ing. Benedikt Aumeier

(jdrewes@tum.de)
72

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Chair of Urban Water Systems Engineering

Technical University of Munich

Prof. Dr.-Ing. Jörg E. Drewes

• Nearly all surface water treatment facilities use filtration

Cholera Outbreak Hamburg 1892

William Philipps Dunbar

• Trinkwasserverordnung (TrinkwV) 2023

• U.S. EPA - Surface Water Treatment Rule

Common Filtration Processes

– Rapid filtration – most common in Europe and the United States

– Slow sand filtration – small utilities / high quality feed water

– Pre-coated filtration – specific applications (green sand filtration)

Comparison between Granular Filtration

Rapid Granular Media Filtration

Rapid Filtration – Process Flow

– Backwash cycle – accumulated material is flushed from the system

• Physical treatment process to remove colloidal particles

• Removal mechanisms during filtration

• If the ratio of particle diameter to media

• If the ratio is smaller, straining is not possible and

• For typical rapid filtration, straining is limited to

• Particles with a density significantly greater

• Insignificant in water treatment!

• Particles that pass the collector surface by a

• Diffusion is most significant for particles

Removal Mechanisms During Filtration

2. Inertia, sedimentation: particle B deviates from the

3. Diffusion: particle C collides with collector due to

particle diameter < 80 µm

• Lowest efficiency for particles of 1-2 µm

Principal Attraction Forces

I decrease in filtration rate, filter performance, and

Slow Sand Filter

Slow Sand Filter

Slow Sand Filter

• River bank filtration (RBF)

Rapid and High-Rate Sand Filters

• Hydraulic loading rate ~120 m3/m2·day (rapid)

• Hydraulic loading rate >240 m3/m2·day (high-rate)

• Designed for removals throughout the entire depth of

• Most of these filters have multiple layers

Filters Classified by Driving Force

Rapid Filtration – Pretreatment

• Destabilization of particles will reduce repulsion forces between

Classification of Rapid Filtration

Classification of Rapid Filtration

• Materials for Rapid-Filtration

Media design parameters

• effective size (d10)

Example 1: Determine ES and UC

Rapid Filtration – Typical Media

Filter Media Selection

Filter Support Media and Underdrains

6. Hydraulics of Flow Through Granular

Hydraulics of Flow Through Granular Media

• Steady, laminar flow

• 100 < Re < 600-800 : Transition flow

• Re > 800 : Turbulent flow

Ranges of Reynolds Number in Granular

Headloss Through Clean Granular Filters

• Insignificant in slow sand and pre-coat

Headloss Through Clean Granular Filters

• Not taking into account media properties…