CVEN 9857

Wastewater Treatment

Biological Nutrient Removal

Professor MJ Manefield

Metcalf & Eddy (2015) 5th edition

• Chapter 7 “Fundamentals of biological treatment”

Content
•Introduction
•Learning objectives
•Some important definitions
•Nitrogen removal
» Nitrification
» Denitrification
» Anammox
•Phosphate removal
•Summary
Introduction
• Excess supply of nutrients can result in
deterioration of water quality leading to the
growth of algal blooms
• Eutrophication can have an adverse economic
impact on water bodies due to loss of
recreational use, fish kill events and increased
potable water treatment costs
• Discharge from WWTP can contribute significant
quantities of nitrogen and phosphorus to rivers,
lakes and reservoirs
Learning objectives
• Understand the need for nutrient removal
processes in wastewater treatment
• Describe the basic parameters underpinning
nitrification and denitrification in wastewater
treatment
• Understand the principles of Enhanced Biological
Phosphate Removal (EBPR)
• Be familiar with the process configurations used
to design for N and P removal
Carbonaceous constituents in sewage
• C organic matter measured by BOD or COD
• TCOD = rbCOD + sbCOD + nbsCOD + nbpCOD

Metcalf Eddy (2015)

Nitrogenous constituents in raw
sewage Nitrite N
Total oxidised N
(1 mg/L) Nitrate N

Total Nitrogen
Ammonia N
Total Kjeldahl N (TKN) (60-70% of TKN)

(50-60 mg/L)
Organic N

Biodegradable Non-Biodegradable

soluble particulate soluble particulate

 Ammonia and Ammonium

In wastewater there is very little ammonia because the pH is close to neutral.

It is mostly the ammonium ion we are dealing with, though people say ammonia.
 Definition of ‘anoxic’ and ‘anaerobic’
• People use these terms in different ways.

1. Anoxic refers to absence of oxygen. Anaerobic processes happen

under anoxic conditions. ‘Anaerobic’ describes processes. ‘Anoxic’
Science

describes conditions. In this usage, there is no such thing as an

anaerobic environment.

2. The term ‘anoxic’ can also be used to describe oxygen free conditions
in which nitrate is present. Used this way, we can have aerobic, anoxic
Engineering

and anaerobic conditions.

Aerobic – with oxygen
Anoxic – without oxygen but with nitrate
Anaerobic – oxygen and nitrate free environment
Nitrogen removal processes
We are mostly talking about ammonium removal
processes because that is the main nitrogen
containing compound in wastewater
There are two processes:

1. Nitrification plus denitrification (established)

2. Anammox (developing)
 Nitrification

Conversion of Ammonia to Nitrate

Aerobic
 Nitrification (ammonium and nitrite oxidation)
• Nitrification is mostly achieved in two stages with the
conversion of ammonia to nitrite by Ammonium Oxidising
Bacteria (AOB) and Archaea (AOA) followed by conversion
of nitrite to nitrate by Nitrite Oxidising Bacteria (NOB)

1 Ammonium oxidation: 2NH4+ + 3O2  2NO2- + 4H+ + 2H2O

2 Nitrite oxidation: 2NO2- + O2  2NO3-

• The first step (ammonium oxidation) is rate limiting and

therefore nitrite concentrations do not build up

Nitrification: 2NH4+ + 4O2  2NO3- + 4H+ + 2H2O

4.57 g O2 is required to oxidise 1 g N (as ammonium)

 Ammonium oxidizing bacteria and archaea
Examples

Nitrosomonas europaea
Aerobic (Oxygen is electron acceptor)
Chemolithotrophic (autotrophic)
Bacteria

Slow growing
Low affinity ammonia monooxygenase (AmoA)

Nitrosopumilus maritimus
Aerobic (Oxygen is electron acceptor)
Chemolithotrophic (autotrophic)
Archaea

Slow growing
High affinity ammonia monooxygenase (AmoA)
Nitrite oxidizing bacteria
Examples

Nitrospira muscoviensis
Aerobic (Oxygen is electron acceptor)
Energy source is nitrite or hydrogen
Mixotrophic for carbon (autotrophic and heterotrophic)
Enzyme responsible – nitrite oxidoreductase

Nitrobacter winogradskyi
Aerobic
Chemolithotrophic (autotrophic)
Enzyme responsible – nitrite oxidoreductase
Comammox (complete ammonia oxidizer)
In 2015 Nitrospira species were discovered to carry
out both ammonia oxidation and nitrite oxidation
The first step in nitrification (ammonium oxidation) produces protons
(acid!)

pH is a measure of proton concentration

2NH4+ + 3O2  2NO2- + 4H+ + 2H2O

2NO2- + O2  2NO3-

Protons react with bicarbonate

NH4+ + 2HCO3- + 2O2  NO3- + 2CO2 + 3H2O

So ammonium oxidation consumes alkalinity

AS conditions for nitrification

Parameter BOD Removal Nitrification

Only

Dissolved oxygen 0.5 - 1.0 1.5 - 2.0

(mg/L)
HRT (h) 4-6 6 - 10
Sludge age (d) 4-6 6 - 10
Low temperature Tolerant Not tolerant
Nitrification growth kinetics
• Growth of nitrifiers can be described by the
Monod equation
• As nitrifiers are slower growing than heterotrophs,
they are more sensitive to the sludge age of
suspended growth (activated sludge) type
reactors
• The effect of substrate concentration, pH,
dissolved oxygen and temperature on growth rate
can be predicted and used to determine required
sludge age
Growth rate () of ammonium oxidising bacteria
according to temperature and ammonium and oxygen concentration

Where:
max = maximum specific growth rate
T = temperature
DO = dissolved oxygen
pHopt = pH 7.2
[NH4+ - N] = effluent ammonia concentration
(Barnes and Bliss, 1983)
Conditions that inhibit nitrification
• Cold temperature
• Deficiencies in key nutrients
• High influent ammonium ion concentration
• Inhibition/Toxicity by metals and other toxins
• Alkalinity
• pH swing or extreme pH (optimum pH 7.2)
• Short retention times
• Low dissolved oxygen concentration
 Denitrification

Conversion of Nitrate to Nitrogen gas (N2)

Anaerobic
Denitrification
• Denitrification is an anaerobic respiration activity
and involves the microbial reduction of nitrate to
nitrite, and ultimately nitrite to nitrogen gas
• Nitrate and nitrite replace oxygen for microbial
respiration
• Some nitrate is removed via incorporation into
biomass (assimilatory removal) rather than via
nitrogen gas (dissimilatory removal)
• Nitrate assimilation: nitrate is reduced to
ammonia for nutrient uptake purposes.
Conditions for denitrification
•No oxygen
•Nitrate / nitrite present
•Carbon source (organic)

Denitrification removes BOD/COD as well as nitrate

Denitrifying bacteria use nitrate as an electron
acceptor and reduce it to nitrogen gas via nitrite,
nitric oxide and nitrous oxide
NO3-  NO2-  NO  N2O  N2
Denitrifying bacteria
• Aerobic conditions – bacteria use O2 as final
electron acceptor in degrading organic carbon
• Anoxic conditions – bacteria use NO-3 as final
electron acceptor in degrading organic carbon
Achromobacter Bacillus
Acinetobacter Chromobacterium
Agrobacterium Corynebacterium
Alcaligenes Hypomicrobium
Arthrobacter Paracoccus
Flavobacterium Pseudomonas
 Process configuration for denitrification
Anoxic zone after the aeration basin
NH3 to NO3 NO3 to N2

• All the BOD is removed before denitrification so organic carbon

has to be added to provide carbon and energy for denitrification
• Denitrifying sludge flocs trap nitrogenous gasses so it doesn’t
settle well in the clarifier/settling/sedimentation basin
 Process configuration for denitrification
Anoxic zone before the aeration basin
NO3 to N2 NH3 to NO3

• Need a nitrate feed loop

• BOD in influent wastewater is used for denitrification so no need to add
acetate or methanol
• Trapped gasses are released with agitation in the aeration basin so the
sludge settles well in the sedimentation basin
Anoxic zone
Denitrification stoichiometry
Carbon and energy for denitrifying bacteria can come from the
reduced organic carbon (BOD) in the wastewater or from reduced
organic carbon substrates like acetate and methanol.
2NO3- + 10e- + 12H+  N2 + 6 H2O

Wastewater
C10H19O3N + 10NO3-  5N2 + 10CO2 + 3H2O + NH3 + 10OH-
Methanol
5CH3OH + 6NO3-  3N2 + 5CO2 + 7H2O + 6OH-
Acetate
5CH3COOH + 8NO3-  4N2 + 10CO2 + 6H2O + 8OH-
Amount of carbon for denitrification

• Amount of organic carbon depends on the

system operating conditions and the type of
electron donor used
• A general rule of thumb is that 4 g of BOD is
needed per g of NO3 reduced
• Alternatively, The McCarty Equation can be used
to determine the amount of carbon required as
an electron donor for denitrification
The McCarty Equation

Cm = 2.47 NO + 1.53 N1 + 0.87 DO

Where:
Cm = required methanol concentration (mg/l)
NO = initial nitrate-nitrogen concentration (mg/l)
N1 = initial nitrite-nitrogen concentration (mg/l)
DO = initial dissolved oxygen concentration (mg/l)
Suspended growth denitrification

Parameter Pre- Post- Combined

denitrification denitrification Pre and Post

Sludge age (d) 6 – 30 6 – 30 8 – 40

HRT (h)
1st anoxic 2 – 5 - 2–5
1st aerobic 4 – 12 4 – 12 4 – 12
2nd anoxic - 2–5 2–5
Reaeration - - 0.5 – 1.0
Design MLSS and recycles
Parameter Pre- Post- Combined
denitrification denitrification Pre and Post
denitrification

MLSS (mg/l) 1,500 – 4,000 1,500 – 4,000 2,000 – 5,000

ML recycle (2 - 4) Q - (4 – 6) Q

RAS 1:1 1:1 1:1

Energy for mixing

Parameter Pre- Post- Combined Pre and Post

denitrification denitrification denitrification

Anoxic (W/m3) 4 – 10 4 – 10 8 – 14

Aerobic (W/m3) 20 – 40 20 – 40 20 - 40
Environmental effects
•Temperature
- bigger impact <20oC
•pH
- similar to aerobic heterotrophs
- optimum pH 6.0 – 8.0
•Inhibitors
- more robust than nitrifiers
- similar to aerobic heterotrophs
 Anammox
Conversion of Ammonia to N2
Anaerobic
 Anammox (Anaerobic ammonia oxidation)
Instead of organic carbon serving as the electron donor,
ammonia is the electron donor and nitrite is the electron
aceptor so there is no need to add oxygen or organic carbon

NH4+ + NO2− → N2 + 2H2O

• Ideal for wastewater with very high NH4- and low BOD
• Eg. wastewater from anaerobic digester
• Anaerobic autotrophs
• Brocadia anammoxidans

Anammox bacteria form tight

granules with great settling
properties
Nitrogen cycle
N2

Ni
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Nitrate NO3-

Anammox
Ammonium NH4

Nitrite NO2-

Nitrification
 Anammox plants
High nitrogen removal
No methanol dosing is needed
for denitrification
Up to 60% reduction of power
consumption
Minimal production of excess
sludge
Up to 90% reduction of CO2
First full scale installed in 2002 emission
Up to 50% less space required
Biological Phosphate Removal
Biological phosphate removal
• Conventional aerobic biological processes
remove approximately 40% of influent
phosphorus from municipal sewage
» improved to > 90% by addition of chemical precipitants
e.g., aluminum sulfate
• Enhanced biological phosphorus removal
(EBPR) is possible mechanism involving
Anaerobic Respiration
Conditions for EBPR
• Need an anaerobic / no nitrate stage
• Anaerobic stage has to be followed by aerobic
stage (or can have an anoxic stage in
between)
• Phosphate release in anaerobic stage,
‘excess’ uptake in aerobic stage
EBPR mechanism
Anaerobic Aerobic

Soluble [P]

UPTAKE
RELEASE

Time
EBR - configuration

Anaerobic Aerobic

Air

Modified Activated Sludge

EBPR - PAOs
VFAs PO43- CO2 + H2O PO43-

Energy
Poly-P Anaerobic to Energy
Aerobic Phase Poly-P

PHA
PHA
Recycle
Sludge

O2
Wastage
Anaerobic Aerobic

PAOs (Polyphosphate Accumulating Organisms)

P - transformation
• In the anaerobic zone, bacteria remove soluble
BOD, in particular short chain fatty acids such as
acetate from the influent wastewater concomitant
with release of phosphate into solution
• Increase in phosphate is the result of the
hydrolysis of intracellular polyphosphate to
provide energy for short chain fatty acids
transport
P - transformation (II)
• Upon entering the aerobic stage the stored short
chain fatty acids (as PHB) are used as an energy
source to stimulate uptake of extracellular
phosphate and subsequent storage as
polyphosphate
• The uptake of phosphate during the aerobic
stage is in excess during the anaerobic stage
resulting in substantial overall removal of
phosphorus from the influent wastewater
Volatile fatty acids (VFAs)
• Volatile fatty acids, sometimes short chain fatty
acids (SCFA), or readily biodegradable COD
(rbCOD)
» acetate, butyrate, propionate, valerate
• Not all types of sewage will be suitable for EBPR,
as the influent BOD/COD needs to be at a high
enough concentration to ensure that the VFA:P
ratios are sufficient to stimulate EBPR
microorganisms
Glycogen accumulating organisms

• GAOs have a very similar metabolism to PAOs

but without the ability to take up phosphates in
excess of normal metabolic requirements
• GAOs compete with the PAOs and DPAOs,
thereby inhibiting EBPR
GAOs
VFAs CO2 + H2O

Energy
Glycogen Anaerobic to
Aerobic Phase Glycogen

PHA
PHA
Recycle
Sludge

O2
Anaerobic Aerobic

GAO (Glycogen accumulating organisms)

Bio P essentials
• P release requirement
» Anaerobic zone
» Needs VFA (i.e. rbCOD)
» Low DO
» Adequate HRT
» Low nitrite (anoxic zone essential)
• P uptake requirements
» Aerobic conditions
» Adequate HRT
Common barriers - EBPR
• Insufficient VFA availability for P release
• Insufficient readily degradable COD (rbCDO) for
rapid denitrification
• Return of excessive nitrate concentrations to
anaerobic zone
• Large number of variants for BNR process
» mostly to control nitrate return to anaerobic zones as a
result of weak sewages
Bio P monitoring
• Flow readings
• MLSS
• Sludge settlement
• Sludge blanket levels in clarifier
• Final effluent
• Phosphate concentration exiting the anaerobic
zone
Bio P removal
Advantages
•Low operation cost (OPEX)
» little or no chemical costs
» saving on aeration energy
•Small extra sludge production (10%)
•Usually P and N removal
Bio P removal
Disadvantages
• Difficult to achieve < 1mg/l P.
• Susceptible to weak sewage (e.g. Combined
sewers)
• P is easily re-released from sludge.
• Struvite formation (crystals) and blockage
• More complex to operate
Phosphate removal practices
•Biological
•Chemical
• Choice of chemical (ferric)
• Mixing-flocculation
• Control systems
• Sludge route
• Residual metals
•Combined chemical and biological
Chemical P removal
Chemical P removal
Chemical P removal
Advantages
• Low capital cost retrofit
• Stable sludge (P-locked up)
• Simple to operate !
• Sewage strength not important
• Pre-precipitation reduces BOD
• Easily controlled
BNR process configurations
•3-stage BNR
•5-stage BNR (modified Bardenpho)
•University of Capetown (UCT)
•A2/O process
•Virginia Initiative Plant (VIP)
•JHB (Johannesburg) process
•etc
 3-Stage BNR
Anaerobic Anoxic Aerobic Zones
Primary Secondary
Zone Zone
Clarifier Clarifier

Mixed Liquor Recirculation

Return Activated Sludge

To sludge Processing Waste

Activated
Sludge
3-Stage BNR
Johannesburg modification

Primary Anaerobic Anoxic Aerobic Zones Secondary

Clarifier Zone Zone Clarifier

Mixed Liquor Recirculation

Return Activated Sludge

Pre Anoxic
Zone if required

Waste
To sludge Processing Activated
Sludge
5 Stage (modified Bardenpho)

Anoxic Aerobic Anoxic Aerobic

Anaerobic Clarifier
(re-aeration)
Mixed Liquor Recycle

Return Activated Sludge

Modified University Cape Town

Anaerobic Anoxic Anoxic Aerobic Clarifier

Mixed Liquor Recycle (nitrate)

Return Activated Sludge

Mixed Liquor Recycle (no nitrate)

 Seboking configuration
Enlarged Anaerobic
Anoxic Zone
Primary Zone Secondary
Clarifier VFA Clarifier
Stepped Feed Supplement
Aerobic Zones

Return Activated Sludge

Waste
To sludge Processing Activated
Sludge
Anaerobic zone (VFA addition)
 3 Stage with Sidestream Fermenter

Primary Anaerobic Anoxic Aerobic Zones Secondary

Clarifier Zone Zone Clarifier

Mixed Liquor Recirculation

Liquors

Return Activated Sludge

Raw Sludge Fermenter

if required
Waste
Activated
To sludge Processing
Sludge
Sidestream Fermenter
Bio P sludge quality
P removal - summary
Chemical P
•Small to medium STW (<50,000 pe)
•Fixed film processes ( BAF/SAF/RBC)
•Weak waste water systems
Biological P
•Large STW (> 100,000 pe)
•Nitrogen standards
•Aeration energy saving
•Combined chemical and biological
 Fate of Phosphorus (*)
Solublisation Reprecipitation Dewatering Liquours

ferrous phosphate

Anaerobic Calcium Phosphate

Conditions
pH 7-7.3 Residual PO4
PO4-- Struvite
BNR Struvite precipitation
Potential
SAS Residual soluble Mg
Mg++

K+
HCO3 from digestion
Wastewater
Calcite
Inorganics Ca, Mg, CO3 Residual soluble Calcium Precipitation
Potential

SAS = surplus activated sludge (= WAS)

Struvite (Mg NH4 PO4.6H2O)
•Magnesium ammonium phosphate (MAP)

•Drivers for Phosphorus Removal from liquors

» Reduce nuisance struvite precipitation
» Reduce recycled P loads - increase COD:P ratio on
BNR plant
» Potential For Product Recovery - may recoup some of
liquor treatment costs
Struvite Formation

• Struvite = MgNH4PO4.6H2O
• Mole to mole to mole ratio = 1:1:1
• High concentrations of ammonium, phosphate and
magnesium
• High pH & alkalinity
• High conductivity
• Low temperature
Struvite precipitation
Control or limit the formation of
struvite
•Alum addition (to precipitate P)
•Ferric/ferrous chloride addition (to precipitate P)
•Ferric sulfate addition (to precipitate P, potential release
of H2S during digestion process)
•Lime addition (to precipitate P and adjust alkalinity)
•Anti-scalent chemical addition (to suppress struvite
formation)
•pH reduction with acid or CO2 dosing
•High pressure water jets to flush (routine maintenance
and recycle)
Struvite recovery

• Add excessive
Settlement Reaction Settlement MgCl2
Zone Zone Zone
• Increase pH
• Air

De-sludged struvite
Air
Phosphorus recovery

Struvite based
fertiliser
Notes
•Nitrogen removal basics
•Nitrogen removal design
•Biological phosphate removal
Reading
Jetten et al. (2002) Improved nitrogen removal by
application of new nitrogen-cycle bacteria.
Re/Views in Environmental Science and
Bio/Technology 1, 51-63
Van der Star et al. (2007) Startup of reactors for
anoxic ammonium oxidation: Experiences from the
first full-scale anammox reactor in Rotterdam. Water
Research
Van Eekert et al. (2012) Explorative research on
innovative nitrogen recovery. STOWA report 51
References
Janssen P.M.J., Meinema K. and van der Roest
H.F. (2002) Biological Phosphorus Removal. Water
and Wastewater Practitioner Series: STOWA report.
IWA Publishing, London
van Dongen L.G.J.M., Jetten M.S.M. and van
Loosdrecht M.C.M. (2001) The Combined
Sharon/Anammox Process. Water and Wastewater
Practitioner Series: STOWA report. IWA Publishing,
London