Simulating 2,4,6-trinitrotoluene (TNT) elimination in a pond inhabited by freshwater algae of the Rhizoclonium genus

Keywords: Rhizoclonium; explosives; 2,4,6-trinitrotoluene; aquatic ecosystems; biodegradation; decontamination

Abstract Military operations over large areas of Ukraine lead to release of explosives and their derivatives into the environment with subsequent accumulation in natural and artificial water bodies, which unwittingly serve as reservoirs for collecting pollutants from the catchment area. The need to restore aquatic ecosystems dictates the search for efficient, cost-effective and environmentally friendly methods for the elimination of explosives, which corresponds to the processes of biological treatment. In this work, we examined the ability of common freshwater algae of the genus Rhizoclonium to detoxify 2,4,6-trinitrotoluene (TNT) under model conditions of water pollution (at a TNT concentration of 100 mg/L). The exposure time of the algae to TNT was 48 hours, during which the content of TNT and nitrites in the aqueous medium was monitored, as well as the content of chlorophyll and the activity of glutathione S-transferase in plant tissues. 2,4,6-trinitrotoluene was extracted from the aqueous medium with toluene, followed by separation in a separatory funnel, removal of residual water with sodium sulfate, and reduction of the extract volume using a rotary evaporator. The decrease in the concentration of TNT, established by GC-MS technique, was 66.4% by the end of the experiment, while the content of nitrites increased almost 15-fold. In the algae cells, a threefold increase in the enzymatic activity was observed already in the second hour of exposure, followed by a gradual decrease and maintenance at a level of 50% of the control until the end of the experiment. The total chlorophyll content increased significantly from the sixth hour of exposure to the end of the experiment due to an increase in the content of chlorophyll b. The results obtained indicate the efficient biodegradation process and prospects of using algae of the genus Rhizoclonium for cleaning water bodies contaminated with TNT.

References

Adamia, G., Chogovadze, M., Chokheli, L., Gigolashvili, G., Gordeziani, M., Khatisashvili, G., Kurashvili, M., Pruidze, M., & Varazi, T. (2018). About possibility of alga Spirulina application for phytoremediation of water polluted with 2,4,6-trinitrotoluene. Annals of Agrarian Science, 16 (3), 348–351.
Ankit, Bauddh, K., & Korstad, J. (2022). Phycoremediation: Use of algae to sequester heavy metals. Hydrobiology, 1(3), 288–303.
Brentner, L. B., Tanaka, S., Merchie, K. M., Schnoor, J. L., & VanAken, B. (2008). Expression of glutathione S-transferases in poplar trees (Populus trichocarpa) exposed to 2,4,6-trinitrotoluene (TNT). Chemosphere, 73(5), 657–662.
Chugh, M., Kumar, L., Shah, M. P., & Bharadvaja, N. (2022). Algal bioremediation of heavy metals: An insight into removal mechanisms, recovery of by-products, challenges, and future opportunities. Energy Nexus, 7, 100129.
Dietz-Vargas, C., Valenzuela-Ibaceta, F., Carrasco, V., & Pérez-Donoso, J. M. (2023). Solid medium for the direct isolation of bacterial colonies growing with polycyclic aromatic hydrocarbons or 2,4,6-trinitrotoluene (TNT). Archive of Microbiology, 205(7), 271.
Ding, N., Wang, A., Zhang, X., Wu, Y., Wang, R., Cui, H., Huang, R., & Luo, Y. (2017). Identification and analysis of glutathione S-transferase gene family in sweet potato reveal divergent GST-mediated networks in aboveground and underground tissues in response to abiotic stresses. BMC Plant Biology, 17, e225.
Eisentraeger, A., Reifferscheid, G., Dardenne, F., Blust, R., & Schofer, A. (2007). Hazard characterization and identification of a former ammunition site using microarrays, bioassays and chemical analysis. Environmental Toxicology and Chemistry, 26(4), 634–646.
Habig, W. H., & Jakoby W. B. (1981). Assays for differentiation of glutathione S-transferases. Methods in Enzymology, 77, 398–405.
Kalderis, D., Juhasz, A. L., & Boopathy, R. (2011). Steve comfort soils contaminated with explosives: Environmental fate and evaluation of state-of the art remediation processes (IUPAC Technical Report). Pure and Applied Chemistry, 83(7), 1407–1484.
Kröger, M., & Fels, G. (2000). 14C-TNT synthesis revisited. Journal of Labelled Compounds and Radiopharmaceuticals, 43(3), 217–227.
Kumar, A., Nighojkar, A., Varma, P., Prakash, N. J., Kandasubramanian, B., Zimmermann, K., & Dixit, F. (2023). Algal mediated intervention for the retrieval of emerging pollutants from aqueous media. Journal of Hazardous Materials, 455(5), 131568.
Kumar, S., & Trivedi, P. K. (2018). Glutathione S-transferases: Role in combating abiotic stresses including arsenic detoxification in plants. Frontiers in Plant Sciences, 9, 751.
Lachance, B., Renoux, A. Y., Sarrazin, M., Hawari, J., & Sunahara, G. I. (2004). Toxicity and bioaccumulation of reduced TNT metabolites in the earthworm Eisenia andrei exposed to amended forest soil. Chemosphere, 55(10), 1339–1348.
Leal, P., Ojeda, J., Sotomayor, C., & Buschman, A. H. (2020). Physiological stress modulates epiphyte (Rhizoclonium sp.)-basiphyte (Agarophyton chilense) interaction in co-culture under different light regime. Journal of Applied Phycology, 32, 3219–3232.
Lewis, T. A., Newcombe, D. A., & Crawford, R. L. (2004). Bioremediation of soils contaminated with explosives. Journal of Environment Management, 70(4), 291–307.
Li, Y., Wang, D., Xu, X. T., Gao, X. X., Sun, X., & Xu, N. J. (2017). Physiological responses of a green algae (Ulva prolifera) exposed to simulated acid rain and decreased salinity. Photosynthetica, 55(4), 623–629.
Liu, Q., Zhang, G., Ding, J., Zou, H., Shi, H., & Huang, C. (2018). Evaluation of the removal of potassium cyanide and its toxicity in green algae (Chlorella vulgaris). Bulletin of Environmental Contamination and Toxicology, 100(2), 228–233.
Luo, J., Huang, C., Peng, F., Xue, X., & Wang, T. (2017). Effect of salt stress on photosynthesis and related physiological characteristics of Lycium ruthenicum Murr. Acta Agriculturae Scandinavica, Section B – Soil and Plant Science, 67(8), 680–692.
Ma, X., Chen, X., Fan, J., Wang, Y., & Zhang, J. (2022). The response of three typical freshwater algae to acute acid stress in water. Journal of Environmental Science and Health. Part A Toxic/Hazardous Substances and Environmental Engineering, 57(2), 102–110.
Martseniuk, V. M., Prychepa, M. V., & Marenkov, O. M. (2023). Changes of activity of energy and ion exchange enzymes and the energy substrates content in tissues of Perca fluviatilis and Rutilus rutilus under toxic water pollution. Hydrobiological Journal, 59(3), 66–77.
Mercimek, H. A., Dincer, S., Guzeldag, G., Ozsavli, A., Matyar, F., Arkut, A., Kayis, F., & Ozdenefe, M. S. (2015). Degradation of 2,4,6-trinitrotoluene by P. aeruginosa and characterization of some metabolites. Brazilian Journal of Microbiology, 46(1), 103–111.
Morsi, H. H., El-Sabbagh, S. M., Mehesen, A. A., Mohamed, A. D., Al-Harbi, M., Elkelish, A., El-Sheekh, M. M., & Saber, A. A. (2023). Antibacterial activity of bioactive compounds extracted from the egyptian untapped green alga Rhizoclonium hieroglyphicum. Water, 15(11), 2030.
Osuna-Ruiz, I., Salazar-Leyva, J. A., Saiz, C. L., & Burrgos-Hernandes, A. (2019). Enhancing antioxidant and antimutagenic activity of the green seaweed Rhizoclonium riparium by bioassay-guided solvent partitioning. Journal of Applied Phycology, 31, 3871–3881.
Piotrowska-Niczyporuk, A., Czerpak, R., Pietryczuk, A., & Wędołowska, M. (2008). The effect of indomethacin on the growth and metabolism of green alga Chlorella vulgaris Beijerinck. Plant Growth Regulation, 55(2), 125–136.
Polechońska, L., & Klink, A. (2022). Macrophytes as passive bioindicators of trace element pollution in the aquatic environment. Wiley Interdisciplinary Reviews: Water, 10(2), e1630.
Polechońska, L., Klink, A., Dambiec, M., & Rudecki, A. (2018). Evaluation of Ceratophyllum demersum as the accumulative bioindicator for trace metals. Ecological Indicators, 93(4), 274–281.
Rylott, E. L., & Bruce, N. C. (2019). Right on target: Using plants and microbes to remediate explosives. International Journal of Phytoremediation, 21(11), 1051–1064.
Rylott, E. L., Gunning, V., Tzafestas, K., Sparrow, H., Johnston, E. J., Brentnall, A. S., Potts, J. R., & Bruce, N. C. (2015). Phytodetoxification of the environmental pollutant and explosive 2,4,6-trinitrotoluene. Plant Signaling and Behavior, 10(1), e977714.
Saber, A. A., Ichihara, K., & Cantonati, M. (2017). Molecular phylogeny and detailed morphological analysis of two freshwater Rhizoclonium strains from contrasting spring types in Egypt and Italy. Plant Biosystems, 151(5), 800–812.
Salama, E.-S., Roh, H.-S., Dev, S., Ali Khan, M., Abou Shanab, R. A. I., Chang, S. W., & Jeon, B.-H. (2019). Algae as a green technology for heavy metals removal from various wastewater. World Journal of Microbiology and Biotechnology, 35, 74–93.
Senze, M., & Kowalska-Góralska, M. (2019). Evaluation of the bioaccumulation of metals in submerged plants of the Verdon River and Lake Sainte-Croix (France) – preliminary research. Journal of Elementology, 25(1), 297–314.
Serrano-González, M. Y., Chandra, R., Castillo-Zacarias, C., Robledo-Padilla, F., Rostro-Alanis, M. de J., & Parra-Saldivar, R. (2018). Biotransformation and degradation of 2,4,6-trinitrotoluene by microbial metabolism and their interaction. Defence Technology, 14(2), 151–164.
Shemer, B., Yagur-Kroll, S., Hazan, C., & Belkin, S. (2018). Aerobic transformation of 2,4-dinitrotoluene by Escherichia coli and its implications for the detection of trace explosives. Applied and Environmental Microbiology, 84(4), e01729-17.
Snellinx, Z., Nepovím, A., Taghavi, S., Vangronsveld, J., Vanek, T., & van der Lelie, D. (2002). Biological remediation of explosives and related nitroaromatic compounds. Environmental Science and Pollution Research International, 9(1), 48–61.
Subashchandrabose, S. R., Ramakrishnan, B., Megharaj, M., Venkateswarlu, K., & Naidu, R. (2013). Mixotrophic cyanobacteria and microalgae as distinctive biological agents for organic pollutant degradation. Environment International, 51, 59–72.
Wang, L., Fu, H., Zhao, J., Wang, J., Dong, S., Yuan, X., Li, X., & Chen, M. (2023). Genome-wide identification and expression profiling of glutathione S-transferase gene family in foxtail millet (Setaria italica L.). Plants, 12(5), 1138.
Wintermans, J. F. G. M., & De Mots, A. (1965). Spectrophotometric characteristics of chlorophyll a and b and their phaeophytins in etanol. Biochimica et Biophysica Acta, 109(2), 448–453.
Xu, M., He, L., Sun, P., Wu, M., Cui, X., Liu, D., Adomako-Bonsu, A. G., Geng, M., Xiong, G., Guo, L., & Maser, E. (2023). Critical role of monooxygenase in biodegradation of 2,4,6-trinitrotoluene by Buttiauxella sp. S19-1. Molecules, 28(4), 1969.
Yesipova, N., Marenkov, O., Sharamok, T., Nesterenko, O., & Kurchenko, V. (2022). Development of the regulation of hydrobiological monitoring in circulation cooling system of the Zaporizhzhia Nuclear Power Plant. Eastern-European Journal of Enterprise Technologies, 2(10), 6–17.
Zhang, H., Zhong, H., Wang, J., Sui, X., & Xu, N. (2016). Adaptive changes in chlorophyll content and photosynthetic features to low light in Physocarpus amurensis Maxim and Physocarpus opulifolius “Diabolo”. PeerJ, 4, e2125.

Comments (0)

No login
gif