Removal of pathogenic indicator microorganisms during partial nitrification: the role of free nitrous acid

doi:10.1007/s11783-024-1793-6

Front. Environ. Sci. Eng.

2024, Vol. 18

Issue (3) : 33 https://doi.org/10.1007/s11783-024-1793-6

RESEARCH ARTICLE

Removal of pathogenic indicator microorganisms during partial nitrification: the role of free nitrous acid

Jiaojiao Xu¹, Xiaotian Chen², Rui Tang³, Jingwei Feng¹, Shoujun Yuan¹, Wei Wang¹, Zhen-Hu Hu¹(

)

¹. Anhui Engineering Laboratory of Rural Water Environment and Resource, School of Civil Engineering, Hefei University of Technology, Hefei 230009, China
². Hefei 168 High School, Hefei 230025, China
³. Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, Shanghai 200090, China

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Abstract

● 83% ± 13% E. coli and 59% ± 27% Enteroco ccus were removed by partial nitrification.

● FNA exposure leads to surface collapse of E. coli and Enterococcus .

● Bacteria inactivation was due to the breakdown of cell walls and cell membranes.

● Enterococcus was more resistant to FNA treatment than E. coli .

Digested wastewater contains pathogenic microorganisms and high ammonia concentrations, which can pose a potential risk to public health. Effective removal of pathogens and nitrogen is crucial for the post-treatment of digested wastewater. Partial nitrification-anammox is an energy-saving nitrogen removal process. Free nitrous acid (FNA), an intermediate product of partial nitrification, has the potential to inactivate microorganisms. However, the efficiency and mechanisms of FNA-related inactivation in pathogens during partial nitrification remains unclear. In this study, Enterococcus and Escherichia coli (E. coli) were selected to investigate the efficiency and mechanisms of FNA-related inactivation in partial nitrification process. The results revealed that 83% ± 13% and 59% ± 27% of E. coli and Enterococcus were removed, respectively, in partial nitrification process at FNA concentrations of 0.023−0.028 mg/L. When the concentration of FNA increased from 0 to 0.5 mg/L, the inactivation efficiencies of E. coli and Enterococcus increased from 0 to 99.9% and 89.9%, respectively. Enterococcus exhibited a higher resistance to FNA attack compared to E. coli. 3D-laser scanning microscopy (3D-LSM) and scanning electron microscopy (SEM) revealed that FNA exposure caused the surface collapse of E. coli and Enterococcus, as well as visible pore formation on the surface of E. coli cells. 4',6-Diamidino-2-phenylindole dihydrochloride n-hydrate (DAPI)/propidium iodide (PI) and biomolecule leakage confirmed that inactivation of E. coli and Enterococcus occurred due to breakdown of cell walls and cell membranes. These findings indicate that partial nitrification process can be used for the removal of residual pathogenic microorganisms.

Keywords Partial nitrification Free nitrous acid Pathogenic indicator microorganism Inactivation Cell structure

Corresponding Author(s): Zhen-Hu Hu

About author:

Peng Lei and Charity Ngina Mwangi contributed equally to this work.

Issue Date: 14 November 2023

Cite this article:

Jiaojiao Xu,Xiaotian Chen,Rui Tang, et al. Removal of pathogenic indicator microorganisms during partial nitrification: the role of free nitrous acid[J]. Front. Environ. Sci. Eng., 2024, 18(3): 33.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-024-1793-6
https://academic.hep.com.cn/fese/EN/Y2024/V18/I3/33

Tab.1 Conditions for treatment of E. coli and Enterococcus with FNA

Fig.1 Concentrations of FNA, NO₂⁻−N, NO₃⁻−N, and NH₄⁺−N in the partial nitrification SBR reactor.

Fig.2 Removal of E. coli and Enterococcus during the partial nitrification process. (a) E. coli; (b) Enterococcus. (E. coli_inf: cell density of E. coli in the influent; E. coli_eff: cell density of E. coli in the effluent; Enterococcus_inf: cell density of Enterococcus in the influent; Enterococcus_eff: cell density of Enterococcus in the effluent).

Fig.3 Inactivation efficiencies of E. coli and Enterococcus at various FNA concentrations for 24 h. (a) E. coli; (b) Enterococcus.

Fig.4 3D-LSM images of E. coli and Enterococcus with and without FNA treatment. (a) and (c) E. coli without FNA treatment; (b) and (d) E. coli with FNA treatment; (e) and (g) Enterococcus without FNA treatment; (f) and (h) Enterococcus with FNA treatment.

Tab.2 Arithmetic average roughness and maximum height of E. coli and Enterococcus

Fig.5 SEM images of E. coli and Enterococcus with and without FNA treatment. E. coli without (a) and with (b) FNA treatment; Enterococcus without (c) and with (d) FNA treatment.

Fig.6 Fluorescence staining of E. coli and Enterococcus treated with FNA for 24 h. (a–c) E. coli; (d–f) Enterococcus.

Fig.7 Biomolecule leakage from FNA-treated and FNA-untreated bacteria for 48 h. (a) protein release from E. coli; (b) protein release from Enterococcus; (c) intracellular DNA concentration.

1	A W Abbew, A A Amadu, S Qiu, P Champagne, I Adebayo, P O Anifowose, S Ge. (2022). Understanding the influence of free nitrous acid on microalgal-bacterial consortium in wastewater treatment: a critical review. Bioresource Technology, 363: 127916 https://doi.org/10.1016/j.biortech.2022.127916
2	A Akhiar, A Battimelli, M Torrijos, H Carrere. (2017). Comprehensive characterization of the liquid fraction of digestates from full-scale anaerobic co-digestion. Waste Management, 59: 118–128 https://doi.org/10.1016/j.wasman.2016.11.005
3	O O Alegbeleye, A S Sant’Ana. (2020). Manure-borne pathogens as an important source of water contamination: an update on the dynamics of pathogen survival/transport as well as practical risk mitigation strategies. International Journal of Hygiene and Environmental Health, 227: 113524 https://doi.org/10.1016/j.ijheh.2020.113524
4	M Amiri, G A Crawford, J C Earthman. (2021). Quantitative percussion diagnostics for evaluating porosity and surface roughness of cold sprayed and laser deposited materials. Journal of Materials Research and Technology, 14: 312–323 https://doi.org/10.1016/j.jmrt.2021.06.047
5	APHA (2005). Standard Methods for the Examination for Water and Wastewater, 21st ed. Washington, DC: APHA, AWWA and WEF
6	X Bai, P M Schenk, Z Yuan, P A Lant, S Pratt. (2015). Enhanced triacylglyceride extraction from microalgae using free nitrous acid pre-treatment. Applied Energy, 154: 183–189 https://doi.org/10.1016/j.apenergy.2015.04.045
7	S Borowski, J Domanski, L Weatherley. (2014). Anaerobic co-digestion of swine and poultry manure with municipal sewage sludge. Waste Management, 34(2): 513–521 https://doi.org/10.1016/j.wasman.2013.10.022
8	H Chen, M Zhang. (2013). Effects of advanced treatment systems on the removal of antibiotic resistance genes in wastewater treatment plants from Hangzhou, China. Environmental Science & Technology, 47(15): 8157–8163 https://doi.org/10.1021/es401091y
9	M Chislett, J Guo, P L Bond, A Jones, Z Yuan. (2020). Structural changes in cell-wall and cell-membrane organic materials following exposure to free nitrous acid. Environmental Science & Technology, 54(16): 10301–10312 https://doi.org/10.1021/acs.est.0c01453
10	M Chislett, J Guo, P L Bond, Y Wang, B C Donose, Z Yuan. (2022). Reactive nitrogen species from free nitrous acid (FNA) cause cell lysis. Water Research, 217: 118401 https://doi.org/10.1016/j.watres.2022.118401
11	M Chislett, J Guo, P L Bond, Z Yuan. (2021). Structural changes in model compounds of sludge extracellular polymeric substances caused by exposure to free nitrous acid. Water Research, 188: 116553 https://doi.org/10.1016/j.watres.2020.116553
12	M Cho, J Kim, J Y Kim, J Yoon, J H Kim. (2010). Mechanisms of Escherichia coli inactivation by several disinfectants. Water Research, 44(11): 3410–3418 https://doi.org/10.1016/j.watres.2010.03.017
13	C Dennehy, P G Lawlor, G E Gardiner, Y Jiang, P Cormican, M S McCabe, X Zhan. (2017). Process stability and microbial community composition in pig manure and food waste anaerobic co-digesters operated at low HRTs. Frontiers of Environmental Science & Engineering, 11(3): 4 https://doi.org/10.1007/s11783-017-0923-9
14	H Duan, S Gao, X Li, N H Ab Hamid, G Jiang, M Zheng, X Bai, P L Bond, X Lu, M M Chislett, S Hu, L Ye, Z Yuan. (2020). Improving wastewater management using free nitrous acid (FNA). Water Research, 171: 115382 https://doi.org/10.1016/j.watres.2019.115382
15	S H Gao, L Fan, Z Yuan, P L Bond. (2015). The concentration-determined and population-specific antimicrobial effects of free nitrous acid on Pseudomonas aeruginosa PAO1. Applied Microbiology and Biotechnology, 99(5): 2305–2312 https://doi.org/10.1007/s00253-014-6211-8
16	Y Guo, Z Luo, J Shen, Y Y Li. (2022). The main anammox-based processes, the involved microbes and the novel process concept from the application perspective. Frontiers of Environmental Science & Engineering, 16(7): 84 https://doi.org/10.1007/s11783-021-1487-1
17	X Han, S Zhang, S Yang, L Zhang, Y Peng. (2020). Full-scale partial nitritation/anammox (PN/A) process for treating sludge dewatering liquor from anaerobic digestion after thermal hydrolysis. Bioresource Technology, 297: 122380 https://doi.org/10.1016/j.biortech.2019.122380
18	D Huffman. (2000). Inactivation of bacteria, virus and Cryptosporidium by a point-of-use device using pulsed broad spectrum white light. Water Research, 34(9): 2491–2498 https://doi.org/10.1016/S0043-1354(00)00014-2
19	C Jiang, S Xu, R Wang, S Zhou, S Wu, X Zeng, Z Bai, G Zhuang, X Zhuang. (2018). Comprehensive assessment of free nitrous acid-based technology to establish partial nitrification. Environmental Science. Water Research & Technology, 4(12): 2113–2124 https://doi.org/10.1039/C8EW00637G
20	G Jiang, O Gutierrez, Z Yuan. (2011). The strong biocidal effect of free nitrous acid on anaerobic sewer biofilms. Water Research, 45(12): 3735–3743 https://doi.org/10.1016/j.watres.2011.04.026
21	G Jiang, A Keating, S Corrie, K O’Halloran, L Nguyen, Z Yuan. (2013). Dosing free nitrous acid for sulfide control in sewers: results of field trials in Australia. Water Research, 47(13): 4331–4339 https://doi.org/10.1016/j.watres.2013.05.024
22	G Jiang, Z Yuan (2013). Synergistic inactivation of anaerobic wastewater biofilm by free nitrous acid and hydrogen peroxide. Journal of Hazardous Materials, 250–251: 91–98 https://doi.org/10.1016/j.jhazmat.2013.01.047
23	M Li, G Song, R Liu, X Huang, H Liu. (2022). Inactivation and risk control of pathogenic microorganisms in municipal sludge treatment: a review. Frontiers of Environmental Science & Engineering, 16(6): 70 https://doi.org/10.1007/s11783-021-1504-5
24	Y Li, H Liu, G Li, W Luo, Y Sun. (2018). Manure digestate storage under different conditions: chemical characteristics and contaminant residuals. Science of the Total Environment, 639: 19–25 https://doi.org/10.1016/j.scitotenv.2018.05.128
25	Y Li, D Wang, G Yang, X Yuan, Q Xu, Q Yang, Y Liu, Q Wang, B J Ni, W Tang, L Jiang. (2020). Enhanced dewaterability of anaerobically digested sludge by in-situ free nitrous acid treatment. Water Research, 169: 115264 https://doi.org/10.1016/j.watres.2019.115264
26	Y Liang, R Ma, A Nghiem, J Xu, L Tang, W Wei, H Prommer, Y Gan (2022). Sources of ammonium enriched in groundwater in the central Yangtze River Basin: anthropogenic or geogenic? Environmental Pollution, 306: 119463 https://doi.org/10.1016/j.envpol.2022.119463
27	M Lin, A Wang, L Ren, W Qiao, S M Wandera, R Dong. (2022). Challenges of pathogen inactivation in animal manure through anaerobic digestion: a short review. Bioengineered, 13(1): 1149–1161 https://doi.org/10.1080/21655979.2021.2017717
28	B Ma, L Yang, Q Wang, Z Yuan, Y Wang, Y Peng. (2017). Inactivation and adaptation of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria when exposed to free nitrous acid. Bioresource Technology, 245: 1266–1270 https://doi.org/10.1016/j.biortech.2017.08.074
29	A Manero, A R Blanch. (1999). Identification of Enterococcus spp. with a biochemical key. Applied and Environmental Microbiology, 65(10): 4425–4430 https://doi.org/10.1128/AEM.65.10.4425-4430.1999
30	G Maniakova, I Salmeron, S Nahim-Granados, S Malato, I Oller, L Rizzo, M I Polo-Lopez. (2021). Sunlight advanced oxidation processes vs ozonation for wastewater disinfection and safe reclamation. Science of the Total Environment, 787: 147531 https://doi.org/10.1016/j.scitotenv.2021.147531
31	G Mao, D Wang, Y Bai, J Qu. (2023). Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: benefits of wastewater treatment plant upgrade. Frontiers of Environmental Science & Engineering, 17(7): 82 https://doi.org/10.1007/s11783-023-1682-4
32	I Ofori, S Maddila, J Lin, S B Jonnalagadda. (2018). Chlorine dioxide inactivation of Pseudomonas aeruginosa and Staphylococcus aureus in water: the kinetics and mechanism. Journal of Water Process Engineering, 26: 46–54 https://doi.org/10.1016/j.jwpe.2018.09.001
33	K Oguma, H Katayama, H Mitani, S Morita, T Hirata, S Ohgaki. (2001). Determination of pyrimidine dimers in Escherichia coli and Cryptosporidium parvum during UV light inactivation, photoreactivation, and dark repair. Applied and Environmental Microbiology, 67(10): 4630–4637 https://doi.org/10.1128/AEM.67.10.4630-4637.2001
34	S Qiu, Z Li, Y Hu, Q Yang, L Chen, R Liu, X Zhan. (2021). N2O generation via nitritation at different volumetric oxygen transfer levels in partial nitritation-anammox process. Journal of Cleaner Production, 293: 126104 https://doi.org/10.1016/j.jclepro.2021.126104
35	R P Schwarzenbach, B I Escher, K Fenner, T B Hofstetter, C A Johnson, U von Gunten, B Wehrli. (2006). The challenge of micropollutants in aquatic systems. Science, 313(5790): 1072–1077 https://doi.org/10.1126/science.1127291
36	V M Vadivelu, Z Yuan, C Fux, J Keller. (2006). The inhibitory effects of free nitrous acid on the energy generation and growth processes of an enriched nitrobacter culture. Environmental Science & Technology, 40(14): 4442–4448 https://doi.org/10.1021/es051694k
37	C A Vergara-Castiblanco, F Freire-Santos, A M Oteiza-Lopez, M E Ares-Mazas. (2000). Viability and infectivity of two Cryptosporidium parvum bovine isolates from different geographical location. Veterinary Parasitology, 89(4): 261–267 https://doi.org/10.1016/S0304-4017(00)00210-7
38	H Wang, J Wang, M Zhou, W Wang, C Liu, Y Wang. (2022). A versatile control strategy based on organic carbon flow analysis for effective treatment of incineration leachate using an anammox-based process. Water Research, 215: 118261 https://doi.org/10.1016/j.watres.2022.118261
39	Q Wang, L Ye, G Jiang, P D Jensen, D J Batstone, Z Yuan. (2013). Free nitrous acid (FNA)-based pretreatment enhances methane production from waste activated sludge. Environmental Science & Technology, 47(20): 11897–11904 https://doi.org/10.1021/es402933b
40	G Wen, Z Liang, X Xu, R Cao, Q Wan, G Ji, W Lin, J Wang, J Yang, T Huang. (2020). Inactivation of fungal spores in water using ozone: kinetics, influencing factors and mechanisms. Water Research, 185: 116218 https://doi.org/10.1016/j.watres.2020.116218
41	J Wu, Q Yang, W Luo, J Sun, Q Xu, F Chen, J Zhao, K Yi, X Wang, D Wang, et al. (2018). Role of free nitrous acid in the pretreatment of waste activated sludge: extracellular polymeric substances disruption or cells lysis? Chemical Engineering Journal, 336: 28–37 https://doi.org/10.1016/j.cej.2017.11.038
42	X Zhan, L Xiao. (2017). Livestock Waste 2016−International Conference on Recent Advances in Pollution Control and Resource Recovery for the Livestock Sector. Frontiers of Environmental Science & Engineering, 11(3): 16 https://doi.org/10.1007/s11783-017-0958-y
43	T Zhang, Q Wang, J Khan, Z Yuan. (2015). Free nitrous acid breaks down extracellular polymeric substances in waste activated sludge. RSC Advances, 5(54): 43312–43318 https://doi.org/10.1039/C5RA06080J
44	Y Zhang, Y Jiang, S Wang, Z Wang, Y Liu, Z Hu, X Zhan. (2021). Environmental sustainability assessment of pig manure mono- and co-digestion and dynamic land application of the digestate. Renewable & Sustainable Energy Reviews, 137: 110476 https://doi.org/10.1016/j.rser.2020.110476
45	Q Zhao, J Ma, I Zeb, L Yu, S Chen, Y Zheng, C Frear. (2015). Ammonia recovery from anaerobic digester effluent through direct aeration. Chemical Engineering Journal, 279: 31–37 https://doi.org/10.1016/j.cej.2015.04.113
46	M Zheng, Z Wang, J Meng, Z Hu, Y Liu, Z Yuan, S Hu. (2021). Inactivation kinetics of nitrite-oxidizing bacteria by free nitrous acid. Science of the Total Environment, 752: 141876 https://doi.org/10.1016/j.scitotenv.2020.141876
47	B W Zhou, S G Shin, K Hwang, J H Ahn, S Hwang. (2010). Effect of microwave irradiation on cellular disintegration of Gram positive and negative cells. Environmental Science & Technology, 87(2): 765–770

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