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Frontiers of Environmental Science & Engineering

ISSN 2095-2201

ISSN 2095-221X(Online)

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Front. Environ. Sci. Eng.    2021, Vol. 15 Issue (2) : 32    https://doi.org/10.1007/s11783-020-1324-z
RESEARCH ARTICLE
Start-up of PN-anammox system under low inoculation quantity and its restoration after low-loading rate shock
Guoliang Zhang1, Liang Zhang1(), Xiaoyu Han2, Shujun Zhang2, Yongzhen Peng1
1. National Engineering Laboratory for Advanced Municipal Wastewater Treatment and Reuse Technology, Engineering Research Center of Beijing, Beijing University of Technology, Beijing 100124, China
2. Research and Development Center of Beijing Drainage Group Technology, Beijing 100124, China
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Abstract

• PN-A was start-up under low inoculation amount and a higher NRR was achieved.

• PN-anammox system was successfully restored by aggressive sludge discharge.

• Increase in granular sludge was the important factor to rapid recovery.

• Enrichment of AOB and AnAOB in granular sludge favors the stable operation.

Partial nitritation (PN)-anaerobic ammonium oxidation (anammox) is a promising pathway for the biological treatment of wastewater. However, the destruction of the system caused by excessive accumulation of nitrate in long-term operation remains a challenge. In this study, PN-anammox was initialized with low inoculation quantity in an air-lift reactor. The nitrogen removal rate of 0.71 kgN/(m3·d) was obtained, which was far higher than the seed sludge (0.3 kgN/(m3·d)). Thereafter, excess nitrate build-up was observed under low-loading conditions, and recovery strategies for the PN-anammox system were investigated. Experimental results suggest that increasing the nitrogen loading rate as well as the concentration of free ammonium failed to effectively suppress the nitrite oxidation bacteria (NOB) after the PN-anammox system was disrupted. Afterwards, effluent back-flow was added into the reactor to control the up-flow velocity. As a result, an aggressive discharge of sludge that promoted the synergetic growth of functional bacteria was achieved, leading to the successful restoration of the PN-anammox system. The partial nitritation and anammox activity were in balance, and an increase in nitrogen removal rate up to 1.07 kgN/(m3·d) was obtained with a nitrogen removal efficiency of 82.4% after recovery. Besides, the proportion of granular sludge (over 200 mm) increased from 33.67% to 82.82%. Ammonium oxidation bacteria (AOB) along with anammox bacteria were enriched in the granular sludge during the recovery period, which was crucial for the recovery and stable operation of the PN-anammox system.
Keywords PN-anammox      Granular sludge      Excess nitrate build-up      Recovery strategy for partial nitrification      Aggressive discharge of sludge     
Corresponding Author(s): Liang Zhang   
Issue Date: 07 September 2020
 Cite this article:   
Guoliang Zhang,Liang Zhang,Xiaoyu Han, et al. Start-up of PN-anammox system under low inoculation quantity and its restoration after low-loading rate shock[J]. Front. Environ. Sci. Eng., 2021, 15(2): 32.
 URL:  
https://academic.hep.com.cn/fese/EN/10.1007/s11783-020-1324-z
https://academic.hep.com.cn/fese/EN/Y2021/V15/I2/32
Phase Operation time (d) HRT (h) Inf ammonia concentration (mg/L) NLR
(kg/(m3·d))		 pH
Inf. Eff.
I 1–94 12 200–500 0.4–1.0 7.9–8.3 7.6–7.8
II 95–133 12 400 0.8 8.4–8.6 7.5–7.9
III 134–152 20 400 0.48 8.0–8.3 7.4–7.6
IV 153–267 8 400 1.2 8.5–8.6 7.6–7.8
Tab.1  The long-term operation condition of reactor
Fig.1  Schematic diagram of the air-lift reactor set-up.
Specificity Pre-denaturation Amplification cycle (40 cycles)
AOB 95°C 30 s 95°C 30 s; 52°C 30 s; 72°C 40 s
Nitrospira 95°C 30 s 95°C 30 s; 52°C 30 s; 72°C 40 s
Nitrobacter 94°C 30 s 94°C 1 min; 50°C 1 min; 72°C 2 min
AnAOB 95°C 3 min 95°C 10 s; 56°C 30 s; 72°C 45 s
Total bacterial 94°C 3 min 94°C 30 s; 57°C 1 min; 72°C 1 min
Tab.2  Amplification procedure and annealing temperature for different strains
Strains Probe Sequence
AOB amoA-F 5′-GGGGTTTCTACTGGTGGT-3′
amoA-R 5′-CCCCTCKGSAAAGCCTTCTTC-3′
Nitrospira NSR1113F 5′-CCTGCTTTCAGTTGCTACCG-3′
NSR1264R 5′-GTTTGCAGCGCTTTGTACCG-3′
Nitrobacter FGPS872f 5′-TTTTTTGAGATTTGCTAG-3′
FGPS1269r 5′-CTAAAACTCAAAGGAATTGA-3′
AnAOB Amx368f 5′-TTCGCAATGCCCGAAAGG-3′
Amx820r 5′-AAAACCCCTCTACTTAGTGCCC-3′
Total bacterial Total bacterial -1055F 5′-ATGGCTGTCGTCAGCT-3′
Total bacterial -1392R 5′-ACGGGCGGTGTGTAC-3′
Tab.3  The 16S rRNA-targeted oligonucleotide probes for different strains
Fig.2  Overall performance of the reactor during continuous operation: (a) nitrogen concentration; (b) NLR(nitrogen loading rate) and NRR(nitrogen removal rate);(c) NRE(nitrogen removal efficiency);(d) DNO3?/DNH4+ ratio;(e) MLSS.
Fig.3  Result of SAA experiment in different phases:(a)phase II(the 127th day);(b)phase III (the 156th day);(c)phase IV (the 210th day).
Fig.4  Variations of the percentage of sludge with different particle size during long-term operation.
Fig.5  Variation of abundance of AnAOB, AOB, Nitrospira and Nitrobacter in different phases.
Sample AOB Nitrospira Nitrobacter AnAOB Total bacterial
D115 floc 8.47 × 1010 2.63 × 109 6.68 × 108 3.23 × 109 7.89 × 1011
D115 granule 1.68 × 1010 1.34 × 109 4.28 × 108 8.63 × 1011 1.59 × 1012
D172 floc 9.42 × 109 1.10 × 1010 4.91 × 107 4.68 × 108 1.33 × 1011
D172 granule 3.40 × 1010 7.39 × 109 2.09 × 108 1.38 × 1011 9.44 × 1011
D220 floc 7.27 × 1010 9.79 × 109 1.23 × 108 1.76 × 109 2.82 × 1011
D220 granule 1.71 × 1010 2.46 × 109 1.31 × 108 1.07 × 1011 2.82 × 1011
D255 floc 3.93 × 1011 8.46 × 109 1.71 × 108 2.54 × 109 5.10 × 1011
D255 granule 2.04 × 1011 8.05 × 109 1.23 × 108 1.55 × 1011 4.83 × 1011
Tab.4  Variation of copy number (copies/gVSS) of AnAOB, AOB, Nitrospira, Nitrobacter and total bacterial in different phases
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