4th IWARR CONFERENCE 2021

Light Irradiation Enables Rapid Start-up of Nitritation through Limiting

nxrB Gene Expression and Stimulating Ammonia-Oxidizing Bacteria
Lingfeng Wanga, Shuang Qiua, Shijian Ge*a
aJiangsu Key Laboratory of Chemical Pollution Control and Resources Reuse, School of Environmental and Biological
Engineering, Nanjing University of Science and Technology, Xiao Ling Wei 200, Nanjing 210094, Jiangsu, China
*Corresponding author: Dr. Shijian Ge, geshijian1221@njust.edu.cn; geshijian1221@126.com

INTRODUCTION
The applications of anaerobic ammonium oxidation Current approaches for nitritation Research objective
(Anammox) or nitritation/microalgae processes for
wastewater treatment have attracted increasing attention,  Temperature a b
due to reduced carbon and aeration consumption, lower  Low dissolved  Free ammonia
greenhouse gas emission and less sludge generation.1, 2 oxygen  Free nitrous
Nitritation is one of the essential step to provide the substrate  pH acid
PLC
nitrite (NO2-), involving complete or partial conversion of  Sludge
PLC  Formic acid
ammonia (NH4+) to NO2- without further conversion to nitrate retention time  Sulfide Figure 2 – Graphical abstract
(NO3-).3 Stable nitritation largely relies on the microbial  ……  ……  To explore the technical feasibility of the rapid achievement of
competition of nitrifiers in which ammonia-oxidizing bacteria nitritation using light irradiation.
Figure 1 – Two kinds of conceptual approaches for nitritation,
(AOB) are preferred to be retained, while nitrite-oxidizing (a) mainstream and (b) sidestream.  To investigate the effects of light irradiation on the activities and
bacteria (NOB) are undesirable. microbial composition changes of AOB and NOB in activated sludge.

METHODS DISCUSSION
4. Microbial dynamics and transcriptional responses to light irradiation.
Responses of AOB and NOB to short-term light irradiation

 Light energy density

𝑊𝑊∙𝑡𝑡
(𝑃𝑃 = 𝑈𝑈 𝑉𝑉∙𝐶𝐶 , kJ/mg VSS
 The ratio of OUR values at light
condition to control (AOB/NOB)
𝑘𝑘∙𝑃𝑃
𝑟𝑟 = 𝑒𝑒𝑒𝑒𝑒𝑒⁡
( + 𝑏𝑏𝑃𝑃2.5 )
2476 .38
Figure 3 – Schematic representation of the batch experiment.

Experimental Bioreactor Set-up and Operations for Start-up of Nitritation.

ONLSBR OFFLSBR
Stirrers

Effluent Effluent

High light
irradiation PLC
board

Aeration pH DO

Feeding tank Side light irradiation

treat unit

Figure 4 – Schematic representation of the SBRs: ONLSBR (on-line direct light

irradiation reactor) and OFFLSBR (off-line indirect light irradiation reactor). Figure 8 – Percent of community abundance of the different families in the samples collected from a) control (cycle 50), b)
ONLSBR (on-line direct light irradiation reactor) (cycle 38), c) OFFLSBR (off-line indirect light irradiation reactor) (cycle
50). d) Venn diagram of AOB OTUs in control, ONLSBR and OFFLSBR. e) The OUT numbers of AOB and NOB (left) and
new AOB genus obtained in ONLSBR and OFFLSBR (right). f) The numbers of key functional genes (amoA and nxrB)
related to ammonia and nitrite oxidation in ONLSBR, OFFLSBR and control.

RESULTS 5. The energy consumption contrast of CNDM and NDM.

1. Experimental and modelling responses 2. Both Online and Offline Light

of AOB and NOB to light energy density. Irradiation Help Achieving Nitritation.
Phase 1 Phase 2 Phase 3 Phase 4 Phase 5
a 20 0.08
80
Nitrogenous compound

16 0.06
60
Figure 9 – Nitrogen (red), DO (blue) and
P (kJ/mg VSS)

0.04
carbon (purple) transformation pathways
(mg N/L)

12
NAR (%)

40 0.02
8
in bacteria-microalgae consortium system
20 0.00 with a) conventional nitrification-
4
0
-0.02 denitrification (CNDM) and b)
0 -0.04 nitritation-denitritation (NDM).
-10 0 10 20 30 40 50 60
Phase 5
b 20
Phase 1 Phase 2 Phase 3 Phase 4
0.08
80
Nitrogenous compound

16 0.06
60
P (kJ/mg VSS)

0.04
(mg N/L)

12
NAR (%)

40 0.02
8
20 0.00
4 -0.02
0
Figure 5 – Relative activities of AOB and NOB 0 -0.04
observed in a) 40 W, b) 120 W, c) 200 W light -10 0 10 20 30 40 50 60
Phase 1 Phase 2 Phase 3 Phase 4 Phase 5
irradiation groups in comparison to control group. c 20 NH4+-N
0.08
NO2--N 80

CONCLUSIONS
Nitrogenous compound

The sludge concentrations were controlled at 16 NO3--N

0.06

around 2000 mg VSS/L under room temperature 60

P (kJ/mg VSS)

NAR 0.04
(mg N/L)

12 P
NAR (%)

for all batches. 40 0.02

8
20 0.00  Low light energy density can stimulate AOB, and light irradiation can inhibit NOB by limiting
4
3. AOB and NOB activities changes 0
-0.02
the expression of nxrB gene.
0
in different phases of three SBRs -0.04
-10 0 10 20
Cycle
30 40 50 60
 The P value of 0.03-0.08 kJ / mg VSS can be applied to the establishment of nitritation.
a Control b Control
Figure 6 – Nitrogen removal performance
 Both ONLSBR and OFFLSBR could establish nitritation, and the NAR of ONLSBR reached
5 ONLSBR 6 ONLSBR in a) control, b) ONLSBR (on-line direct
Ammomnium oxidation rate

OFFLSBR OFFLSBR
5 light irradiation reactor) and c) OFFLSBR 70%.
Nitrite oxidation rate

4
(off-line indirect light irradiation reactor).
(mg /g MLVSS/h)

(mg /g MLVSS/h)

4  Long-term light irradiaiton could enrich Nitrosomonadaceae, stimulate the production of

3
3 new AOB, and set up nitritation by inhibiting NOB.
Figure 7 – a) AOB and b) NOB activities
2
2 determined in different phases of control,  The nitritation established by light irradiation can be directly applied to NDM system.
1 1
ONLSBR (on-line direct light irradiation Compared with CNDM system, NDM can save 18.3% oxygen and 26.6% organic carbon
reactor) and OFFLSBR (off-line indirect
0 0 light irradiation reactor and reduce the production of 17% sludge.
0 15 30 45 60 0 15 30 45 60
Cycle Cycle

References:
[1] Peng, Y.; Zhu, G. 2006 Biological nitrogen removal with nitrification and denitrification via nitrite pathway. Appl. Microbiol. Biotechnol. 73, (1), 15-26.
[2] Wang, M.; Yang, H.; Ergas, S. J.; van der Steen, P. 2015 A novel shortcut nitrogen removal process using an algal-bacterial consortium in a photo-sequencing batch reactor (PSBR). Water Res. 87, 38-48.
[3] Ge, S.; Wang, S.; Yang, X.; Qiu, S.; Li, B.; Peng, Y. 2015 Detection of nitrifiers and evaluation of partial nitrification for wastewater treatment: A review. Chemosphere. 140, 85-98.

Acknowledgement: The authors would like to acknowledge the support of the National Natural Science Foundation of China (52000103 and 51708294), Natural Science Foundation of Jiangsu Province for Distinguished Young
Scholars (BK20190022), Natural Science Foundation of Jiangsu Province (BK20180497 and BK20181303), and the Fundamental Research Funds for the Central Universities (30921011219 and 30920021117). Dr. Shijian Ge
acknowledges the support of Distinguished Professorship of Jiangsu Province, China and China Association for Science and Technology.

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