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DOI 10.1007/s00253-016-8058-7
MINI-REVIEW
&
* Yeshi Cao
cao_yeshi1949@hotmail.com
* Mark C. M. van Loosdrecht
M.C.M.vanLoosdrecht@tudelft.nl
Introduction
Motivations
Microbial anoxic ammonium oxidation (anammox) was first
discovered in Delft in the 1980s (Mulder 1989) and regarded
as Bone of the most startling ones in environment
biotechnology^ (Rittmann and McCarty 2001). It is a remarkable microbial conversion process: ammonium is oxidized
using nitrite, as electron acceptor, to produce nitrogen gas by
anaerobic ammonium-oxidizing bacteria (anammox bacteria
(AnAOB)), which belongs to the order of Planctomycetales
(Strous et al. 1999). The partial nitritation and anammox (PN/
A) (also termed deammonification) process consists of two
consecutive reactions: ammonium is partially oxidized to nitrite aerobically (nitrite shunt or nitrogen shortcut) by
ammonium-oxidizing bacteria (AOB), and subsequently, the
remaining ammonium reacts with nitrite to form nitrogen gas
anaerobically by AnAOB (van Loosdrecht 2008). Compared
to conventional biological nitrogen removal (nitrification/denitrification), PN/A as an autotrophic nitrogen removal process has three unique advantages (Jetten et al. 1997; Wett
2007; Daigger 2014): (i) 60% of reduction in oxygen demand
for partial oxidation of ammonium to nitrite, (ii) nearly 100%
elimination of carbon demand for denitrification, and (iii) 80%
reduction of excess sludge.
Process/reactor/
/biomass
Single-stage PN/A
OLAND RBC
Single-stage PN/A
MBBR (K3 carriers,
10-mm thickness of biofilm)
MBBR (Carriers K1)
IFAS
PN/A MBBR (K1 carriers)
with argumentation by carrier
transfer
PN/A granular and biofilm
Li et al. (2016)
Single-stage PN/A
OLAND RBC
0.001
0.12
46
16
4.0
2.3
0.0025
40
24
>48
0.001
0.2
1.8
1
21
0.0036
0.0036
2228
25
15
17
20
10
29
15
25
30
20
10
1.52.0 1723
5.57.2 20
15
10
18
0.027
7.3
pH
7.37.5
77.5
NA
7.27.5
7.3
7.57.7
7.3
7.07.3
NA
7.3
7.2
Ambient NA
3.7
0.2
12
T
(C)
0.0029
11
HRT
(h)
0.005
Reactor
volume
(m3)
0.4
0.5
0.5
0.04
0.015
0.005iv
0.01iv
0.12
0.08
0.16
0.85
1.11.4
1.21.4
N.A.
0.02iii
0.02iii
0.20c
0.1
0.22iii
NH4-N 5560
COD/N 0
NH4-N 5560
COD/N 0.52.
NH4-N 50
COD 0
NH4-N 45
COD 4471
NH4-N 28
sCOD 56
COD 100
NH4-N 70
Glucose-COD 40
NH4-N 45
% of TN removal 81
NH4-N 4.8
NO2-N 0
NO3-N 8.2
% of TN removal 6580
NH4-N 37
NO2-N 514
NO3-N 1015
% N removal 50
NH4-N 8
NO2-N 15
NO3-N 13
% TN removal40
NH4-N 4
NO2 10
NO3-N 20
NH4-N <8
NO2-N 18
NO3--N 240
% of TN removal 25
% of TN removal 44
% of TN removal
Up to 60
0.200.30
BOD/N 0.67
fed by chemical
enhanced A-stage
NH4-N 6631
COD 0
N.A.
% of TN removal 9589
NO2-N <1
% of TN removal 8973
% of TN removal 47
% of TN removal 73
%TN removal 73
% of TN removal 46
% of TN removal 5070
0.510.49
0.480.41
0.20.1
0.53
0.53
0.270.43
NA
% of TN removal 85
% of N removal 90
No NO2 accumulation
% of TN removal 70
NH4-N 1.4
NO2-N 0.6
NO3-N 5.3
0.9
0.1
0.028b
N.A.
0.14
0.03a
N.A.
NH4-N 44
sCOD/N 0.81.1
NH4-N 190
HAc 100
NH4-N 60160
COD 0
NH4-N 70
COD 0
NH4-N 24
COD/N 13
Influent
(mg/L, C/N)
Summary of the status of mainstream PN/A process research and development as of summer 2016
Reference
Table 1
Reactor
volume
(m3)
0.012
Process/reactor/
/biomass
Two-stage PN/A
Ma et al. (2011)
0.0045
0.0028 (PN)
0.0085
(Anammox)
0.34 (PN)
0.45
(Anammox)
Reference
Table 1 (continued)
9843
10
20
25
3.0
4.0
5.3
2730
30
915
15
2317
1218
2014
T
(C)
4.6
1.4
5.7
5.6
15
NA
2.53
HRT
(h)
7.5
7.58.2
6.7
6.7
NA
6.57.3
NA
NA
NA
pH
0.17iii
COD 80
NH4-N 35
NO2-N 30
COD 30
(NH4 + NO2) 61
PN
NH4-N 33.4
C/N 7
Anammox
NH4-N 5.5
NO2-N 0.7
NO3-N 2.9
0.26
0.027
N.A.
0.15
(PN )
0.05iii (Anammox)
0.25
0.31
0.10
0.11
0.15
0.17
NH4-N 30.8
NO2-N 0.4
NO3-N 0.7
COD/N 7
TN 45.9
COD 44.4
0.15
0.19
0.020.04
N.A.
NH4-N 21
NO2- and NO3-N
<0.4
COD 69
sCOD 46
NH4-N 1540
COD/N 9.2
0.10.3
0.10.3
N.A.
NH4-N 35
sCOD/NH4-N 1.52
NH4-N 2025
sCOD/NH4-N 3
0.12
Influent
(mg/L, C/N)
% of TN removal 81
(NH4 + NO2)-N
0.030.17
% of TN removal 88
NH4-N 0.84
NO2-N 0.61
TN 5.4
% of TIN removal for
shortcut 75
NH4-N 6.6
NO2-N 2.1
NO3-N 4.6
% of TIN for anammox 38
NH4 4.5
NO2-N 0.21
NO3-N 4.0
% TN removal 82
NH4-N 2.7
NO2-N 2.0
NO3-N 1.9
% TN removal 88
NH4-N 1.7
NO2-N 1.2
NO3-N 1.1
% of TIN Removal 70
NH4-N <10
NO3-N <7
NO2-N <2
% of TIN Removal 40
% of TN removal > 70
NH4-N < 2
TN < 10
% of TN removal 6082
0.004
0.012
0.008
Reactor
volume
(m3)
30
16
T
(C)
0.12
0.11
20
30
316.5 29
12.5
0.12
0.28
HRT
(h)
7.9
7.3
NA
pH
NH4-N 50
NO2-N 50
COD 0
COD 10
NH4-N 16.8
NO2-N 20.5
NO3-N 13.9
sCOD 25.5
NH4-N 19
COD 71
sCOD 47
Influent
(mg/L, C/N)
0.64iv
0.63iv
N.A.
NA
% of NO2 removal 94
% of NH4 removal 93
% of NO2 removal 92
% of NH4-N removal 79
0.47 (non-substrate N.A.
limited)
0.05 (non-substrate
limited)
0.46iv
% of TN removal 74
0.48 iv
% of TN removal 78
5.72
2.28
Calculated according to the biofilm based removal rate and filling ratio
Calculated according to the biofilm-based nitrogen loading and carrier filling ratio
Calculated according to the influent NH4, reactor volume, and removal efficiency
HRT hydraulic retention time, TN total nitrogen, TIN total inorganic nitrogen, SBR sequence batch reactor, HAc acetate, N.A. not available, EGSB expanded granular sludge bed, OLAND
oxygen-limited autotrophic nitrogen denitrification, RBC rotating biofilm contactor, IFAS integrated fixed-film and activated sludge, MBBR moving bed biofilm reactor, UASB up-flow
anaerobic sludge blanket
Anammox granular
sludge UASB
Process/reactor/
/biomass
Ma et al. (2013)
Reference
Table 1 (continued)
the conventional denitrification significantly (75% of TN removal via nitrite path, Wett et al. 2015; 27.1% of TN removal
through conventional denitrification for Changi WRP, Cao
et al. 2014) due to the high influent C/N ratio (712 g COD/
g N) (Regmi et al. 2014a; Cao et al. 2013). In general, the
results of warm anammox are more promising compared with
those of cold anammox. The performance of some laboratory
PN/A systems such as granular sludge reactor operating at
18 C (Winkler et al. 2012a) and hybrid moving bed biofilm
reactor (MBBR) operating at 15 C (Laureni et al. 2016) still
achieved efficient TN removal (> 70%). Pilot-scale and prototype PN/A (reactor volume 50 m3) processes, fed with low
C/N influent ratio after the carbon pretreatment process, have
achieved effective TN removal (7050%) and NRR (0.1
0.3 kg N/m3 day) at temperature between 23 and 17 C
(Lotti et al. 2014b; Geilvoet et al. 2015; Veuillet et al. 2015).
The performance of the latter is similar to or even higher than
that (0.050.10 kg N/m3 day) in conventional nitrification/
denitrification process (ATV 1997). The reduction of TN removal efficiency of pilot-scale process compared to those of
laboratory- scale process reflects the scale-up effects.
Augmentation proved to be effective in NOB suppression
and TN removal (Veuillet et al. 2015), but neither of the detail
mechanisms involved, design made, and operational guidelines adopted was reported. The performance and NRR deteriorate mainly below 15 C. Hendrickx et al. (2014), Lotti et al.
(2014a), Gilbert et al. (2014a), Gao et al. (2015), and Lackner
et al (2015) reported that both effluent nitrite and nitrate concentrations increased and NRR was insignificant at approximately 10 C due to the drastic decrease of anammox activity
approx. below 15 C (Dosta et al. 2008; Gilbert et al. 2014a;
Lotti et al. 2015a) and relatively active NOB at low temperatures (Isanta et al. 2015; Ma et al. 2015a). The studies on final
polishing to remove residual ammonium, nitrate, the side
product of anammox, and nitrite are also at its infancy and
in absence of which large amount of disinfectant is expended
in reuse application such as NEWater production in
Singapore. In summary, the progress achieved so far demonstrates the feasibility of mainstream PN/A process applications even under low temperature, but more research and development are still needed.
This review paper attempts to update the readers with the
overview of the current state of research and development of
mainstream PN/A process, i.e., what has been achieved in
knowledge and applications in microbiology and process
technology and what are reaming as the bottlenecks for
scale-up for full-scale application. Following the brief introduction, the microbiology knowledge generated on key microbial groups (AOB, NOB, AnAOB, and HB) involved in
the PN/A process and their dynamics of interaction are described. Also, the effects of influent C/N ratio on the process
performance are discussed. The suppression of NOB through
kinetics and other factors is discussed and analyzed using
practical cases after that in detail. Then, the major characteristics and performance of the different process (single-stage
and two-stage processes) and the reactor configurations
(suspended sludge, biofilm, and hybrid) implemented to date
are analyzed. The final section highlights the key areas of
further studies required to unblock the bottlenecks and to promote successful scale-up for full-scale application of mainstream PN/A process in wastewater treatment plants.
NOB
AOB
O2
NH4+ + O2 NO2-
NO2- + O2 NO3NO2
NH4+
O2
NO2
NH4+ + NO2- N2 +
NO3-
AnAOB
O2
O2 + COD CO2
NO3- + COD
CO2 + N2
NO2- + COD
HB
Nitrosomonas
Nitrosomonas europaea/eutropha
KO
(mg/L)
0.3
NA
KNH4
max
Remarks
(mg N/L)
(per day)
1.3
0.77
20 C, average of literature data
NA
2.1
Pure culture
Nitrosomonas
0.03
0.5
0.54
Nitrosomonas europaea
Nitrosomonas europaea lineages ML1
Nitrosomonas oligotropha
Lineages NL7
Nitrosospira
0.16
0.24
1.22
0.75
1.62
0.48
1.37
0.66
0.77
30 C, enriched culture
30 C, enriched culture, intrinsic kinetics
Hunik (1993)
Park and Noguera
(2007)
NA
0.64
NA
Schramm et al.
(1999)
Nitrosospira
NA
NA
0.70.9
Pure culture
Siripong and
Rittmann (2007)
NOB
KO (mg/L) KNO2
max
Remarks
(mg N/L)
(per day)
Literature
Nitrobacter agilis
Nitrobacter
Nitrobacter
Nitrobacter
0.54
1. 1
NA
0.43
NA
NA
NA
NA
0.84
1.08
1.39
0.67
30 C, enriched culture
20 C, average of literature data
NA
1923 C, enriched culture, intrinsic kinetics
Hunikz. (1993)
Wiesmann (1994)
Prosser (1989)
Blackburne et al.
(2008b)
Nitrobacter
NA
1.713
1.94.0
AOB
Literature
Wiesmann (1994)
Siripong and
Rittmann (2007)
1923 C, enriched culture, intrinsic kinetics Blackburne et al.
(2008b)
Nitrospira
NA
0.41.2
0.51.2
Nitrospira
0.54
0.9
0.25
Nitrospira
NA
0.16
NA
Schramm et al.
(1999)
NA not available
(dpH/dt) (Yang et al. 2007). In addition, the failures of mechanical equipment such as blowers (Veuillet et al. 2015) or
pumps (Lotti et al. 2014b) resulted in serious operational disruption, illustrating the importance of key equipment reliability for a stable mainstream PN/A process.
In many cases, the suppression of NOB is the complexed
integrated effect of multiple factors (Stinson et al. 2013; Cao
et al. 2013; Wu et al. 2016). This increases the difficulties in
the understanding and analysis of the mechanisms involved
and is an area requiring further studies. Given that the PN/A
process occurs at the aerobic/anoxic/anaerobic interphases,
the potential involvement of ammonia-oxidizing archaea
(AOA) (Park et al. 2006; Daigger and Littleton 2013),
comammox (Holger et al. 2015,) and mixotrophic nitrification
and denitrification (Stein 2011; Bellucci et al. 2011; Fitzgerald
et al. 2015) should not be excluded from the scope of the studies.
Augmentation
Further studies
It can be seen that the major bottlenecks for the scale-up of
mainstream PN/A process are (i) unstable performance of the
carbon concentrating pretreatment, (ii) suppression of NOB
especially under low temperatures (1510 C), (iii) the low
activity of AnAOB at low temperatures, and (iv) final
polishing. Thus, the areas for further studies are proposed
accordingly.
Carbon concentrating pretreatment process
More investigations should be carried out on the mechanisms
of the carbon absorption/storage, particularly bio-flocculation/
coagulation; minimization of carbon mineralization; and the
relationships between the influent characteristics and HRAS/
CEPT process efficiency under typical site operational conditions including SRT, HRT, DO, and temperatures, etc. The
real-time control system and facilities to deal with variety of
influent characteristics resulting from seasonal variations and
heavy rain events should be developed and implemented.
Deepening of understanding competition
between autotrophic nitrogen-converting organisms
Expanding the previous studies and models by accommodating the competition between AOB (mainly Nitrosomonas) and
NOB (mainly Nitrospira and Nitrobacter), revisits and further
studies on the wide range of kinetic parameters for AOB,
NOB, and AnAOB in the literature are desired and crucial.
The investigation of intrinsic kinetics and stoichiometry of
Nitrospira at low temperatures can be an appropriate starting
point. The potential involvement of ammonia-oxidizing archaea (AOA), comammox, and mixotrophic nitrifiers and denitrifiers should not be excluded from the scope of the studies.
Intensification of biofilm anammox activity
Intensification of anammox biofilm activity can be conducted
through increase of population, cell activity of AnAOB, and
integrity of biofilm, which helps to increase the anammox
activity and NRR of mainstream PN/A at low temperature.
To achieve these objectives, the following studies are proposed: (i) to utilize larger size of granular sludge or thicker
biofilm; (ii) to improve the immobilization (granulation) process to culture biofilm with higher anammox population, density, and cell activity, including exploring the feasibility of
using fast-growing free/flocculent AnAOB as seeds; and (iii)
to develop effective operational strategies for augmentation
implemented during the winter season. The possible structure
change of granular sludge (and other types of biofilm) under
low temperature should be studied. The impact of temperature
on the activity of AnAOB and how it can be influenced by the
Compliance with ethical standards The article does not contain any
studies with human participants or animals performed by any of the
authors.
Conflict of interest The authors declare that they have no conflict of
interest.
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