Biological Treatment for Nitrogen

and Phosphorus Removal

Dr. Akepati S. Reddy
Associate Professor, Thapar University, Patiala
Head, Deptt. Analytical Services, TCIRD, Patiala
PUNJAb -147004
 Units for Advanced Treatment
• Meant for the removal of residual constituents found in the
treated effluent
• Residual constituents may include
– Inorganic and organic suspended and colloidal solids
– Dissolved organic matter (rectory compounds, volatile
organic compounds, surfactants, pharmaceutical
compounds, etc.) depleting oxygen, proving toxic or
carcinogenic, causing foaming problems, interfering with
coagulation, etc.
– Dissolved inorganic matter like
• Ammonia (oxygen depleting, toxic and chlorine demanding)
• Nitrate (eutrophicating and methamoglobinemia causing)
• Phosphorus (eutrophicating,and interfering with coagulation and
lime soda softening)
– Biological contamination (bacteria, protozoa, viruses, etc.)
causing disease
• Unit operations and processes employed include
– Chemical precipitation, filtration, membrane processes
(reverse osmosis and electrodialysis), adsorption, ion-
exchange process, advanced oxidation and chemical
oxidation processes, air stripping, distillation, etc.
 Organic-N

decomposition
and hydrolysis
assimilation Organic-N
Ammonical-N Organic-N
(net growth)

nitrification lysis & auto-oxidation

oxygen

Nitrite-N

nitrification
oxygen

denitrification
Nitrate-N Nitrogen gas

Organic carbon

Scheme of Biological Nitrogen Removal

 Biological Nitrification
• Wastewaters have organic-N and ammonical-N
– Ammonical-N DO depleting and toxicity causing
– Nitrogen removal needed for controlling eutrophication and
for recharging ground water
– Nitrate nitrogen limits in drinking water (45 mg/l as nitrate
and 10 mg/l as nitrogen)
• Nitrification is a 2-step process (NH3-N to NO2-N to NO3-
N) and usually achieved along with BOD removal in the
same biological treatment unit
• When concentration of potentially toxic and inhibitory
substances is high, two-sludge systems, each with an
aeration tank and a clarifier, in series are used
– First system is for BOD removal (operated at shorter SRT)
– Nitrification occurs in the 2nd system – raw influent is partially
bypassed into the 2nd system to facilitate sufficient
flocculation & clarification – have much longer HRT & SRT
• In attached growth reactors nitrification is accomplished
after BOD removal (sometimes in separate reactor)
 Biological Nitrification

BOD removal nitrification

• Aerobic autotrophic bacteria are responsible for

nitrification
– Nitroso bacteria (Nitrosomonas, Nitrosococcus, Nitrosospira,
Nitrosolobus, Nitrosorobrio, etc.) are responsible for step-1
– Nitro bacteria (Nitrobacter, Nitrococcus, Nitrosdpira,
Nitrospina, Nitroeystis, etc.) are responsible for step-2
 Biological Nitrification
• Oxygen requirement for complete oxidation of ammonia is
4.57 g/g of N (3.43 for nitrite production and 1.14 g for
nitrate production)
• Use of ammonia as nutrient by the autotrophs reduce the
oxygen requirement to 4.25 g/g
• Requires alkalinity of 7.14 g as CaCO3 per g of nitrogen
(7.07g if use of ammonia as nutrient is considered)
• Nitrification is inhibited by
– Low DO levels (<0.5 mg/l is inhibitory - rate increases with
DO upto 3 to 4 mg/l)
– pH below 6 is inhibitory and 7.5 to 8 is optimal
– Nitrifiers are sensitive to a multitude of organic toxicants
(solvents, amines, proteins, tannins, phenols, alkohols,
cyanates, ethers, carbamates, benzene, etc.)
– Metals are inhibitory (complete inhibition at 0.25 mg/l for
nickel and chromium, and 0.1 mg/l for copper)
– Unionized ammonia can also be inhibitory
 Biological Denitrification
• Denitrification: reduction of nitrate by heterotrophic
bacteria to nitric oxide, nitrous oxide, and nitrogen gas
– Wide range of bacteria (but not algae and fungi) are capable
– Certain autotrophic bacteria under anaerobic conditions and
certain heterotrophic nitrifying bacteria under aerobic
conditions are also capable of converting ammonia into
nitrogen gas
• Nitrate removal occurs in assimilating or dissimilating
modes
– Assimilative mode involves reduction to ammonia for use in
cell synthesis (occurs if ammonical-N is not available)
– Dissimilative mode is biological denitrification and is coupled
with respiratory electron transport chain
• Oxygen equivalence of using nitrate or nitrite in place of
xoygen is 2.86 g/g for nitrate and 1.71 g/g for nitrite
• Alkalinity is produced in the denitrification process (3.57 g
of alkalinity (as CaCO3) per gram of nitrate reduced
• Denitrification is inhibited at higher DO levels (>0.2 mg/l
for pseudomonas & 0.13 mg/l for highly dispersed growth)
 Biological Denitrification
• Two basic schemes of ASP are used for denitrification
– Known as substrate denitrification or preanoxic
denitrification
• anoxic tank followed by an aeration tank
• organic matter of the influent acts as electron donor
– Known as postanoxic denitrification (occurs after nitrification)
• Endogenous decay of microbial mass supplies electron donor
• Often exogenous carbon source (methanol or acetate) is used
• Rate of reduction is relatively slower
• Either a suspended growth or an attached growth system can
be used for the denitrification
• Granular medium filter can be used both for the denitrification
and for the filtration removal of suspended solids
• Nitrification & denitrification can also occur simultaneously
– Nitrification on the floc surface (if DO in the bulk liquid is high
enough) and denitrification in the floc interior (if DO in the
interior is low enough)
– Depending on the mixing conditions, nitrification and
denitrification can occur in the same tank
• Nitrogen level controls substrate utilization kinetics only
 Anoxic Aerobic
section section

Aerobic Anoxic
section section

Biological Denitrification
 Biological Phosphorus Removal
Involves use of Phosphorus Accumulating Organisms (PAO)
in an anaerobic – aerobic system
• Phosphorus is incorporated into sludge (as polyphosphate) in
volutin granules and removed through sludge wastage
Under anaerobic conditions
• Proliferation of PAOs occur
• PAO assimilate fermentation products (specially acetate) into
storage products (polyhydroxybutyrate-PHB) and concomitantly
release stored polyphosphate as orthro phosphate
• Acetate is essential for forming PHB and for providing
competitive advantage to the PAOs
• Presence of nitrate can be inhibitory (acetate can be depleted
and become not available to PAOs)
 Biological Phosphorus Removal
Under aerobic conditions
• stored products (PHB) are oxidized to release energy and in
return phosphate get stored within the cell as polyphosphate
• Molar ratios for Mg, K and Ca of 0.71, 0.5 and 0.25 to phosphorus
respectively are believed to facilitate the polyphosphate storage
• pH and DO should be >6.5 and >1.0 mg/L respectively
Reactor for phosphorus removal is comprised of an anaerobic tank
with HRT 0.5 to 1 hour and placed ahead of an aeration tank
• Return activated sludge and influent are brought in contact in
the anaerobic tank
• Requires >2.5 days SRT in anaerobic and aerobic systems
Typical microbial biomass has 1.5 to 2% phosphorus - in PAOs
phsophorus content can be as high as 20-30%
Stoichiometrically about 10 grams of bCOD is needed for the
removal of one gram of phosphate from wastewater
PAOs form very dense, good settling flocs in the activated sludge
Anaerobic Aenobic
system System
 Biological removal of toxic and
recalcitrant organic substances
Generation time for bacteria is < 20 minutes to a few days
Bacterial growth in a batch reactor can show the following phases:
• When t=0, substrate and nutrients will be in excess and
microbial populations will be very small
• Lag phase:
– acclimation of bacteria to new environment (changed salinity, pH,
temperature, etc.) occurs
– induction of enzyme production takes place
– biomass concentration may be too low to measure
• Exponential growth:
– Bacterial cells multiply at their maximum rate
– Biomass growth curve shows exponential increase
– Only temperature must be influencing the growth
• Stationary phase: biomass concentration remains relatively
constant with time
• Death phase: substrate has been depleted, due to death
biomass concentration experiences exponential decrease