The effects of phenanthrene (Phe) on the denitrification activity and denitrifying genes (narG, nirS and nosZ) were evaluated by dose-response experiments in sediments of Dagu River Estuary (DRE) and Jiaozhou Bay (JZB). The results showed that potential denitrification activity (PDA), N2O, NO3- and NO2- reduction rates of both areas were inhibited with an increase of Phe concentrations. The PDA, N2O, NO3- and NO2- reduction rates of both areas was highest and lowest in the control (DRE: 0.453, 0.427, 7.439 and 3.222mgNkg-1T-1, JZB: 0.592, 0.555, 8.470 and 3.793mgNkg-1h-1) and highest Phe amended treatments (DRE: 0.069, 0.001, 4.486, and 1.563mgNkg-1h-1; JZB: 0.114, 0.024, 5.527 and 2.200mgNkg-1h-1). The inhibition rate of PDA was highest, follow by NO2- reduction and then NO3- reduction. Moreover, with the increasing of Phe concentrations, total bacteria count and the abundance of denitrifying genes were decreased. And N2O accumulation was promoted with the addition of Phe for both areas. Based on the comparison of EC50 values, denitrifiers harboring three genes were more sensitive to Phe than PDA, and denitrifiers harboring nirS gene were more sensitive, followed by nosZ gene, and then narG gene. Furthermore, according to correlation analysis, the relative abundance of denitrifying genes was much more positively correlated with PDA, NO2- and NO2- reduction than total bacteria count. In addition, the denitrification activity and total bacteria count in JZB were more inhibited than that of DRE. This study is useful for understanding the impact of Phe pollution on denitrification in estuary and marine sediments, with profound implications for the management of aquatic ecosystems regarding eutrophication (N-removal) and greenhouse effect.
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Alburquerque, J. A., Sánchez-Monedero, M. A., Roig, A., and Cayuela, M. L., 2015. High concentrations of polycyclic aromatic hydrocarbons (naphthalene, phenanthrene and pyrene) failed to explain biochar’s capacity to reduce soil nitrous oxide emissions. Environmental Pollution, 196 (196C): 72–77.
Bodelier, P., Libochant, J. A., Blom, C. W. P. M., and Laanbroek, H. J., 1996. Dynamics of nitrification and denitrification in root-oxygenated sediments and adaptation of ammonia-oxidizing bacteria to low-oxygen or anoxic habitats. Applied and Environmental Microbiology, 62 (11): 4100–4107.
Cao, Y., Green, P. G., and Holden, P. A., 2008. Microbial community composition and denitrifying enzyme activities in salt marsh sediments. Applied and Environmental Microbiology, 74 (24): 7585–7595.
Canfield, D. E., Glazer, A. N., and Falkowski, P. G., 2010. The evolution and future of Earth’s nitrogen cycle. Science, 330 (6001): 192–196.
Cannarsa, S., Abete, M. C., Zanardi, M., and Squadrone, S., 2014. Polycyclic aromatic hydrocarbons (PAH) in marine sediment of the northwestern Mediterranean Sea (Italy). Journal of Black Sea/Mediterranean Environment, 20 (2): 137–141.
Chon, K. M., and Cho, J. W., 2015. Abundance and expression of denitrifying genes (narG, nirS, norB, and nosZ) in sediments of wastewater stabilizing constructed wetlands. Environmental Engineering Research, 20 (1): 51–57.
Collado, S., Oulego, P., Alonso, S., and Díaz, M., 2017. Flow cytometric characterization of bacterial abundance and physiological status in a nitrifying-denitrifying activated sludge system treating landfill leachate. Environmental Science and Pollution Research, 24 (26): 21262–21271.
Contreras-Ramos, S. M., Alvarez-Bernal, D., Montes-Molina, J. A., Van Cleemput, O., and Dendooven, L., 2009. Emission of nitrous oxide from hydrocarbon contaminated soil amended with waste water sludge and earthworms. Applied Soil Ecology, 41 (1): 69–76.
Foladori, P., Bruni, L., Tamburini, S., and Ziglio, G., 2010. Direct quantification of bacterial biomass in influent, effluent and activated sludge of wastewater treatment plants by using flow cytometry. Water Research, 44 (13): 3807–3818.
Ghosal, D., Ghosh, S., Dutta, T. K., and Ahn, Y., 2016. Current state of knowledge in microbial degradation of polycyclic aromatic hydrocarbons (PAHs): A review. Frontiers in Microbiology, 7 (386): 1369.
Guo, G. X., Deng, H., Qiao, M., Mu, Y. J., and Zhu, Y. G., 2011. Effect of pyrene on denitrification activity and abundance and composition of denitrifying community in an agricultural soil. Environmental Pollution, 159 (7): 1886–1895.
Guo, G. X., Deng, H., Qiao, M., Yao, H. Y., and Zhu, Y. G., 2013. Effect of long-term wastewater irrigation on potential denitrification and denitrifying communities in soils at the watershed scale. Environmental Science & Technology, 47 (7): 3105–3113.
Hallin, S., Jones, C. M., Schloter, M., and Philippot, L., 2009. Relationship between N-cycling communities and ecosystem functioning in a 50-year-old fertilization experiment. The ISME Journal, 3 (5): 597.
Hammes, F. A., and Egli, T., 2005. New method for assimilable organic carbon determination using flow-cytometric enumeration and a natural microbial consortium as inoculum. Environmental Science & Technology, 39 (9): 3289–3294.
Haritash, A. K., and Kaushik, C. P., 2009. Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): A review. Journal of Hazardous Materials, 169 (1-3): 1–15.
Henry, D., 2006. Geometric Theory of Semilinear Parabolic Equations. Vol. 840. Springer, 1–44.
Heiri, O., Lotter, A. F., and Lemcke, G., 2001. Loss on ignition as a method for estimating organic and carbonate content in sediments: Reproducibility and comparability of results. Journal of Paleolimnology, 25 (1): 101–110.
Holtan-Hartwig, L., Dörsch, P., and Bakken, L. R., 2002. Low temperature control of soil denitrifying communities: Kinetics of N2O production and reduction. Soil Biology and Biochemistry, 34 (11): 1797–1806.
Hou, L., Yin, G., Liu, M., Zhou, J. L., Zheng, Y. L., Gao, J., Zong, H. B., Yang, Y., Gao, L., and Tong, C. F., 2015. Effects of sulfamethazine on denitrification and the associated N2O release in estuarine and coastal sediments. Environmental Science & Technology, 49 (1): 326–333.
Liu, N., Yin, P., Zhu, Z. G., Liu, Q. J., Hou, G. H., Lin, X. H., Yan, D. Y., and Zhang, D. L., 2016. Distribution, sources and ecological risk assessment of PAHs in surface sediments from the Dagu River Estuary in Jiaozhou Bay, China. Marine Environment Science, 35 (6): 831–837.
Liu, Y., Shen, K., Wu, Y., and Wang, G., 2018. Abundance and structure composition of nirK and nosZ genes as well as denitrifying activity in heavy metal-polluted paddy soils. Geomicrobiology Journal, 35 (2): 100–107.
Livak, K. J., and Schmittgen, T. D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25 (4): 402–408.
López-Gutiérrez, J. C., Henry, S., Hallet, S., Martin-Laurent, F., Catroux, G., and Philippot, L., 2004. Quantification of a novel group of nitrate-reducing bacteria in the environment by realtime PCR. Journal of Microbiological Methods, 57 (3): 399–407.
Lu, X., Zhang, T., Fang, H. H. P., Leung, K. M., and Zhang, G., 2011. Biodegradation of naphthalene by enriched marine denitrifying bacteria. International Biodeterioration & Biodegradation, 65 (1): 204–211.
Magalhães, C., Kiene, R. P., Buchan, A., Machado, A., Teixeira, C., Wiebe, W. J., and Bordalo, A. A., 2012. A novel inhibitory interaction between dimethylsulfoniopropionate (DMSP) and the denitrification pathway. Biogeochemistry, 107 (1-3): 393–408.
Maliszewska-Kordybach, B., Klimkowicz-Pawlas, A., Smreczak, B., and Janusauskaite, D., 2007. Ecotoxic effect of phenanthrene on nitrifying bacteria in soils of different properties. Journal of Environmental Quality, 36 (6): 1635–1645.
Marton, J. M., Herbert, E. R., and Craft, C. B., 2012. Effects of salinity on denitrification and greenhouse gas production from laboratory-incubated tidal forest soils. Wetlands, 32 (2): 347–357.
McKenney, D. J., and Vriesacker, J. R., 1985. Effect of cadmium contamination on denitrification processes in Brookston clay and Fox sandy loam. Environmental Pollution Series A, Ecological and Biological, 38 (3): 221–233.
Or, D., Smets, B. F., Wraith, J. M., Dechesne, A., and Friedman, S. P., 2007. Physical constraints affecting bacterial habitats and activity in unsaturated porous media - A review. Advances in Water Resources, 30 (6-7): 1505–1527.
Portet-Koltalo, F., Ammami, M. T., Benamar, A., Wang, H., Le Derf, F., and Duclairoir-Poc, C., 2013. Investigation of the release of PAHs from artificially contaminated sediments using cyclolipopeptidic biosurfactants. Journal of Hazardous Materials, 261 (20): 593–601.
Ribeiro, H., Almeida, C. M. R., Mucha, A. P., Teixeira, C., and Bordalo, A. A., 2013. Influence of natural rhizosediments characteristics on hydrocarbons degradation potential of microorganisms associated to Juncus maritimus roots. International Biodeterioration & Biodegradation, 84 (6): 86–96.
Ribeiro, H., Almeida, C. M. R., Magalhães, C., Bordalo, A. A., and Mucha, A. P., 2015. Salt marsh sediment characteristics as key regulators on the efficiency of hydrocarbons bioremediation by Juncus maritimus rhizospheric bacterial community. Environmental Science and Pollution Research, 22 (1): 450–462.
Ribeiro, H., Mucha, A. P., Azevedo, I., Salgado, P., Teixeira, C., Almeida, C. M. R., Joye, S. B., and Magalhães, C., 2016. Differential effects of crude oil on denitrification and anammox, and the impact on N2O production. Environmental Pollution, 216: 391–399.
Rockne, K. J., and Strand, S. E., 2001. Anaerobic biodegradation of naphthalene, phenanthrene, and biphenyl by a denitrifying enrichment culture. Water Research, 35 (1): 291–299.
Schläppy, M. L., Schöttner, S. I., Lavik, G., Kuypers, M. M., de Beer, D., and Hoffmann, F., 2010. Evidence of nitrification and denitrification in high and low microbial abundance sponges. Marine Biology, 157 (3): 593–602.
Seitzinger, S., Harrison, J. A., Böhlke, J. K., Bouwman, A. F., Lowrance, R., Peterson, B., Tobias, C., and Drecht, G. V., 2006. Denitrification across landscapes and waterscapes: A synthesis. Ecological Applications, 16 (6): 2064–2090.
Shao, M. F., Zhang, T., Fang, H. H. P., and Li, X., 2011. The effect of nitrate concentration on sulfide-driven autotrophic denitrification in marine sediment. Chemosphere, 83 (1): 1–6.
Sun, F. L., Wang, Y. S., Sun, C. C., Peng, Y. L., and Deng, C., 2012. Effects of three different PAHs on nitrogen-fixing bacterial diversity in mangrove sediment. Ecotoxicology, 21 (6): 1651–1660.
Sørensen, J., 1978. Denitrification rates in a marine sediment as measured by the acetylene inhibition technique. Applied and Environmental Microbiology, 36 (1): 139–143.
Throbäck, I. N., Enwall, K., Jarvis, Å., and Hallin, S., 2004. Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE. FEMS Microbiology Ecology, 49 (3): 401–417.
Trautwein, K., Kühner, S., Wöhlbrand, L., Halder, T., Kuchta, K., Steinbüchel, A., and Rabus, R., 2008. Solvent stress response of the denitrifying bacterium Aromatoleum aromaticum’ strain EbN1. Applied and Environmental Microbiology, 74 (8): 2267–2274.
Weston, N. B., Vile, M. A., Neubauer, S. C., and Velinsky, D. J., 2011. Accelerated microbial organic matter mineralization following salt-water intrusion into tidal freshwater marsh soils. Biogeochemistry, 102 (1-3): 135–151.
Wang, L., Zheng, B., Nan, B., and Hu, P., 2014. Diversity of bacterial community and detection of nirS- and nirK- encoding denitrifying bacteria in sandy intertidal sediments along Laizhou Bay of Bohai Sea, China. Marine Pollution Bulletin, 88 (1-2): 215–223.
Wilson, S. C., and Jones, K. C., 1993. Bioremediation of soil contaminated with polynuclear aromatic hydrocarbons (PAHs): A review. Environmental Pollution, 81 (3): 229–249.
Wu, Y. Y., Wu, Q. H., Huang, S., Ye, J. X., Zhang, H. J., and Zhang, R. D., 2012. Effect of polycyclic aromatic hydrocarbons on the vertical distribution of denitrifying genes in river sediments. Huanjing Kexue, 33 (10): 3592–3597.
Wuebbles, D. J., 2009. Nitrous oxide: No laughing matter. Science, 326 (5949): 56–57.
Xu, F., Qiu, L., Cao, Y., Huang, J., Liu, Z., Tian, X., Li, A. C., and Yin, X., 2016. Trace metals in the surface sediments of the intertidal Jiaozhou Bay, China: Sources and contamination assessment. Marine Pollution Bulletin, 104 (1-2): 371–378.
Yan, J., Wang, L., Fu, P. P., and Yu, H., 2004. Photomutagenicity of 16 polycyclic aromatic hydrocarbons from the US EPA priority pollutant list. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 557 (1): 99–108.
Yang, X., Ye, J., Lyu, L., Wu, Q., and Zhang, R., 2013. Anaerobic biodegradation of pyrene by Paracoccus denitrificans under various nitrate/nitrite-reducing conditions. Water, Air, & Soil Pollution, 224 (5): 1578.
Yu, H., 2002. Environmental carcinogenic polycyclic aromatic hydrocarbons: photochemistry and phototoxicity. Journal of Environmental Science and Health, Part C, 20 (2): 149–183.
Zeng, W., Zhang, J., Wang, A., and Peng, Y., 2016. Denitrifying phosphorus removal from municipal wastewater and dynamics of ‘Candidatus Accumulibacter’ and denitrifying bacteria based on genes of ppk1, narG, nirS and nirK. Bioresource Technology, 207: 322–331.
Zhang, X. Z., Xie, J. J., and Sun, F. L., 2014. Effects of three polycyclic aromatic hydrocarbons on sediment bacterial community. Current Microbiology, 68 (6): 756–762.
Zhang, Z., Lo, I. M. C., Zheng, G., Woon, K. S., and Rao, P., 2015. Effect of autotrophic denitrification on nitrate migration in sulfide-rich marine sediments. Journal of Soils and Sediments, 15 (4): 1019–1028.
Zhu, J., Wang, Q., Yuan, M. D., Tan, G. Y. A., Sun, F. Q., Wang, C. X., Wu, W. X., and Lee, P. H., 2016. Microbiology and potential applications of aerobic methane oxidation coupled to denitrification (AME-D) process: A review. Water Research, 90: 203–215.
Zhou, H. W., Wong, A. H. Y., Richard, M. K., Park, Y. D., Wong, Y. S., and Tam, N. F., 2009. Polycyclic aromatic hydrocarbon-induced structural shift of bacterial communities in mangrove sediment. Microbial Ecology, 58 (1): 153–160.
Zhou, Z. F., Yao, Y. H., Wang, M. X., and Zuo, X. H., 2017. Coeffects of pyrene and nitrate on the activity and abundance of soil denitrifiers under anaerobic condition. Archives of Microbiology, 199 (8): 1091–1101.
This work was supported by the National Major Project of Water Pollution Control and Management Technology in China (No. 2013ZX07202-007).
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Sun, P., Bai, J., Li, K. et al. Impact of Phenanthrene on Denitrification Activity and Transcription of Related Functional Genes in Estuarine and Marine Sediments. J. Ocean Univ. China 19, 124–134 (2020) doi:10.1007/s11802-020-4006-3
- phenanthrene effect
- denitrifying genes
- estuarine sediment
- marine sediment