Biodegradation of natural and synthetic steroid hormones by river biofilms: Impacts on bacterial community structure

Tatiani Andressa Modkovski; Luis Otávio Miranda Peixoto; Juliane Ribeiro das Chaves; Gabriel Helmer Baer; Rafaela  Imoski; Ludmila da Cruz Souza; Samanta Teixeira Medeiros; Barbara Alves de Lima Nawate; Charles Windson Isidoro Haminiuk; Júlio César Rodrigues de Azevedo

doi:10.5327/Z2176-94782651

Authors

Tatiani Andressa Modkovski Universidade Federal do Paraná (UFPR) - Brazil https://orcid.org/0000-0001-5575-4051
Luis Otávio Miranda Peixoto Universidade Federal do Paraná (UFPR) - Brazil https://orcid.org/0000-0003-4634-6798
Juliane Ribeiro das Chaves Universidade Federal do Paraná (UFPR) - Brazil https://orcid.org/0000-0001-5141-1529
Gabriel Helmer Baer Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0000-0003-1324-8753
Rafaela Imoski Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0000-0002-9665-5794
Ludmila da Cruz Souza Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0009-0001-8084-9500
Samanta Teixeira Medeiros Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0009-0005-2560-0410
Barbara Alves de Lima Nawate Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0000-0003-1842-0532
Charles Windson Isidoro Haminiuk Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0000-0002-1432-9427
Júlio César Rodrigues de Azevedo Universidade Tecnológica Federal do Paraná (UTFPR) - Brazil https://orcid.org/0000-0001-5358-0713

DOI:

https://doi.org/10.5327/Z2176-94782651

Keywords:

bacterial resistance; bioremediation; emerging contaminants; endocrine-disrupting; sampler.

Abstract

Steroid hormones released into the environment by human activities are increasingly found in aquatic environments. Even at low concentrations, they can disrupt natural systems. One of the most affected components is river biofilms, thin layers of microorganisms that grow on submerged surfaces. They play an important role in nutrient cycling, pollutant degradation, and act as indicators of water quality. This study evaluated the ability of natural biofilms from two sites to remove four steroid hormones: natural estrogenic compounds estrone (E1) and 17β-estradiol (E2), synthetic estrogenic compound 17α-ethinylestradiol (EE2), and progesterone (PRO), a natural progestogen. Microcosm assays demonstrated the efficient removal of E1, E2, and EE2, with maximum removal rates of 97.4 (4 days), 93.8 (3 days), and 88.2% (6 days), respectively. PRO was also removed, but with lower efficiency (48.7% in 3 days). The predominant mechanism used by biofilms for hormone removal was biodegradation. Hormone exposure affected biofilms differently depending on their origin. Biofilms collected upstream showed reduced diversity and richness, indicating sensitivity. In contrast, those collected near a wastewater treatment plant had higher diversity and evenness, suggesting resilience and possible adaptation to micropollutants. The predominant phyla were Proteobacteria and Firmicutes, with a reduction in Alloprevotellaand an increase in Paenibacillus following exposure. These results highlight the dual role of river biofilms as natural barriers and biological indicators of contamination by steroid hormones, as well as reinforce their relevance in natural attenuation and the potential for bioremediation in urban rivers.

Downloads

Download data is not yet available.

References

Barcellos, D.S.; Bollmann, H.A.; Azevedo, J.C.R., 2019. Priorização de fármacos nos rios urbanos: o caso dos contraceptivos orais na bacia do rio Belém, Curitiba/PR, Brasil. Revista Ambiente e Água, v. 9 (3), 445-458. https://doi.org/10.4136/1980-993X.

Battin, T.J.; Besemer, K.; Bengtsson, M.M.; Romani, A.M.; Packmann, A.I., 2016. The ecology and biogeochemistry of stream biofilms. Nature Reviews Microbiology, v. 14 (4), 251-263. https://doi.org/10.1038/nrmicro.2016.15.

Bayode, A.A.; Emmanuel, S.S.; Sanni, S.O.; Olalekan, O.A.; Ore, O.T.; Koko, D.T.; Omorogie, M.O., 2024. Current status and performance evaluation of emerging advanced remediation techniques for the removal of steroidal hormones in water. Environmental Chemistry and Ecotoxicology, v. 6, 315-337. https://doi.org/10.1016/j.enceco.2024.07.006.

Bilal, M.; Barceló, D.; Iqbal, H.M.N., 2021. Occurrence, environmental fate, ecological issues, and redefining of endocrine disruptive estrogens in water resources. Science of the Total Environment, v. 800, 18-26. https://doi.org/10.1016/j.scitotenv.2021.149635.

Boopathy, R., 2000. Factors limiting bioremediation technologies. Bioresource Technology, 74 (1), 63-67. https://doi.org/10.1016/s0960-8524(99)00144-3.

Brugnari, T.; Braga, D.M.; Santos, C.S.A.; Torres, B.H.C.; Modkovski, T.A.; Haminiuk, C.W.I.; Maciel, G.M., 2021. Laccases as green and versatile biocatalysts: from lab to enzyme market⠴an overview. Bioresources And Bioprocessing, v. 8 (1), 131. https://doi.org/10.1186/s40643-021-00484-1.

Carles, L.; Wullschleger, S.; Joss, A.; Eggen, R.I.L.; Schirmer, K.; Schuwirth, N.; Stamm, C.; Tlili, A., 2021. Impact of wastewater on the microbial diversity of periphyton and its tolerance to micropollutants in an engineered flow-through channel system. Water Research, v. 203, 117486. https://doi.org/10.1016/j.watres.2021.117486.

Carles, L.; Wullschleger, S.; Joss, A.; Eggen, R.I.L.; Schirmer, K.; Schuwirth, N.; Stamm, C.; Tlili, A.,2022. Wastewater microorganisms impact microbial diversity and important ecological functions of stream periphyton. Water Research, v. 225, 119119. https://doi.org/10.1016/j.watres.2022.119119.

Companhia de Saneamento do Paraná (SANEPAR), 2022. Estudo Técnico e Operacional002 – Almirante Tamandaré/PR. 1-60 (Accessed January 10, 2025) at:. https://www.sanepar.com.br/sites/default/files/Fornecedores/Editais/PPP/00122/Editais/a002_-_almirante_tamandare.pdf

Desiante, W.L.; Minas, N.S.; Fenner, K., 2021. Micropollutant biotransformation and bioaccumulation in natural stream biofilms. Water Research, v. 193, 116846. https://doi.org/10.1016/j.watres.2021.116846.

Ding, N.; Yu, W.; Mo, J.; Rehman, F.; Kasahara, T.; Guo, J., 2024. Does exposure timing of macrolide antibiotics affect the development of river periphyton? Insights into the structure and function. Aquatic Toxicology, v. 275, 107070. https://doi.org/10.1016/j.aquatox.2024.107070.

Du, B.; Fan, G.; Yu, W.; Yang, S.; Zhou, J.; Luo, J., 2020. Occurrence and risk assessment of steroid estrogens in environmental water samples: A five-year worldwide perspective. Environmental Pollution, v. 267, 115405. https://doi.org/10.1016/j.envpol.2020.115405.

Fernandes, G.; Bastos, M.C.; Vargas, J.P.R.; Le Guet, T.; Clasen, B.; Santos, D.R., 2020. The use of epilithic biofilms as bioaccumulators of pesticides and pharmaceuticals in aquatic environments. Ecotoxicology, v. 29 (9), 1293-1305. https://doi.org/10.1007/s10646-020-02259-4.

Fonseca, T.G.; Motta, E.A.; Mass, A.P.; Fongaro, G.; Ramos, F.M.; Machado, M.S.; Bocchese, D.C.F.; Viancelli, A.; Michelon, W., 2021. Toxicity and Enterobacteriaceae Profile in Water in Different Hydrological Events: a case from south brazil. Water, Air, & Soil Pollution, v. 232, 278. https://doi.org/10.1007/s11270-021-05208-x.

Goeury, K.; Munoz, G.; Vo Duy, S.; Prévost, M.; Sauvé, S., 2022. Occurrence and seasonal distribution of steroid hormones and bisphenol A in surface waters and suspended sediments of Quebec, Canada. Environmental Advances, v. 8, 100199. https://doi.org/10.1016/j.envadv.2022.100199.

He, K.; Hain, E.; Timm, A.; Blaney, L., 2021. Bioaccumulation of estrogenic hormones and UV-filters in red swamp crayfish (Procambarus clarkii). Science of the Total Environment, v. 764, 142871. https://doi.org/10.1016/j.scitotenv.2020.142871.

Ide, A.H.; Osawa, R.A.; Marcante, L.O.; Costa Pereira, J.; Azevedo, J.C.R., 2017. Occurrence of Pharmaceutical Products, Female Sex Hormones and Caffeine in a Subtropical Region in Brazil. Clean - Soil, Air, Water, v. 45 (9), 1700334. https://doi.org/10.1002/clen.201700334.

Ilyas, H.; Van Hullebusch, E.D. 2020. A review on the occurrence, fate and removal of steroidal hormones during treatment with different types of constructed wetlands. Journal of Environmental Chemical Engineering, v. 8 (3), 103793. https://doi.org/10.1016/j.jece.2020.103793.

Kaur, R.; Salwan, R.; Sharma, R.; Sharma, V., 2023 Characterization and immobilization of extracellular broad pH active multicopper oxidase from Streptomyces flavomacrosporus for dye-discoloration. Biocatalysis And Agricultural Biotechnology, v. 54, 102955. https://doi.org/10.1016/j.bcab.2023.102955.

Kiel, M.; Engesser, K. 2015. The biodegradation vs. biotransformation of fluorosubstituted aromatics. Applied Microbiology And Biotechnology, v. 99 (18), 7433-7464. https://doi.org/10.1007/s00253-015-6817-5.

Kramer, R.D.; Filippe, T.C.; Prado, M.R.; Azevedo, J.C.R., 2018. The influence of solid-liquid coefficient in the fate of pharmaceuticals and personal care products in aerobic wastewater treatment. Environmental Science and Pollution Research, v. 25 (25), 25515-25525. https://doi.org/10.1007/s11356-018-2609-7.

Li, S.; Zheng, Y.; Wang, C.; Xu, D.; Liu, H.; Song, Y., 2025. Nitrate nitrogen concentrations drive microbial community composition and functional responses in microplastic surface biofilms. Journal of Environmental Chemical Engineering, v. 13 (2), 115504. https://doi.org/10.1016/j.jece.2025.115504.

Li, W.; Luo, D.; Yan, N.; Miao, L.; Adyel, T.M.; Kong, M.; Hou, J., 2023. Effects of polyethylene microplastics with different particle sizes and concentrations on the community structure and function of periphytic biofilms. Journal of Environmental Chemical Engineering, v. 11 (6), 111287. https://doi.org/10.1016/j.jece.2023.111287.

Li, Y.; Zhang, P.; Wang, L.; Wang, C.; Zhang, W.; Zhang, H.; Niu, L.; Wang, P.; Cai, M.; Li, W., 2019. Microstructure, bacterial community and metabolic prediction of multi-species biofilms following exposure to di-(2-ethylhexyl) phthalate (DEHP). Chemosphere, v. 237, 124382. https://doi.org/10.1016/j.chemosphere.2019.124382.

Liang, J.; Li, C.; Mo, J.; Iwata, H.; Rehman, F.; Song, J.; Guo, J., 2024. Metatranscriptomic profiles reveal the biotransformation potential of azithromycin in river periphyton. Water Research, v. 251, 121-140. https://doi.org/10.1016/j.watres.2024.121140.

Lima, B.A.; Modkovski, T.A.; Otavio, L.; Peixoto, M.; Medeiros, S.T.; Baer, G.H., 2025. The influence of hydrological variables on river biofilms: a review of relationships and environmental implications. RBRH, v. 30, e11. https://doi.org/10.1590/2318-0331.302520240083.

Machado, K.S.; Cardoso, F.D.; Azevedo, J.C.R.; Braga, C.B., 2014. Ocorrência de hormônios sexuais femininos na bacia do Rio Iguaçu, Curitiba, Estado do Paraná, Brasil. Acta Scientiarum - Technology, v. 36 (3), 421-427. https://doi.org/10.4025/actascitechnol.v36i3.18477.

Makk, J.; Toumi, M.; Krett, G.; Lange-Enyedi, N.T.; Schachner-Groehs, I.; Kirschner, A.K.T.; Tóth, E., 2024. Temporal changes in the morphological and microbial diversity of biofilms on the surface of a submerged stone in the Danube River. Biologia Futura, v. 75 (3), 261-277. https://doi.org/10.1007/s42977-024-00228-0.

Marques, L.M.T.; Reichert, G.; Cesar, R.M.; Cano, T.C.N.; Azevedo, J.C.R., 2024. Determinação por espectrometria de massas do potencial de bioacumulação de contaminantes emergentes em biofilme. Quim. Nova, v. 47 (10), 1-7. https://doi.org/10.21577/0100-4042.20240071.

Méndez, E., González-Fuentes, M., Rebollar-Perez, G., Méndez-Albores, A., Torres, E. 2016. Emerging pollutant treatments in wastewater: cases of antibiotics and hormones. Journal Of Environmental Science And Health, v. 52 (3), 235-253. https://doi.org/10.1080/10934529.2016.1253391.

Mishra, S.; Huang, Y.; Li, J.; Wu, X.; Zhou, Z.; Lei, Q.; Bhatt, P.; Chen, S.,2022. Biofilm-mediated bioremediation is a powerful tool for the removal of environmental pollutants. Chemosphere, v. 294, 133609. https://doi.org/10.1016/j.chemosphere.2022.133609.

Montagner, C.C.; Sodré, F.F.; Acayaba, R.D.; Vidal, C.; Campestrini, I.; Locatelli, M.A.; Pescara, I.C.; Albuquerque, A.F.; Umbuzeiro, G.A.; Jardim, W.F., 2019. Ten years-snapshot of the occurrence of emerging contaminants in drinking, surface and ground waters and wastewaters from São Paulo State, Brazil. Journal of the Brazilian Chemical Society, v. 30 (3), 614-632. https://doi.org/10.21577/0103-5053.20180232.

Mpupa, A.; Nqombolo, A.; Mizaikoff, B.; Nomngongo, P.N., 2022. Beta-Cyclodextrin-Decorated Magnetic Activated Carbon as a Sorbent for Extraction and Enrichment of Steroid Hormones (Estrone, β-Estradiol, Hydrocortisone and Progesterone) for Liquid Chromatographic Analysis.Molecules, v. 27 (1), 248. https://doi.org/10.3390/molecules27010248.

Ojoghoro, J.O.; Scrimshaw, M.D.; Sumpter, J.P., 2021. Steroid hormones in the aquatic environment. Science of the Total Environment, v. 792, 148-306. https://doi.org/10.1016/j.scitotenv.2021.148306.

Rahman, M.U.; Ullah, M.W.; Shah, J.A.; Sethupathy, S.; Bilal, H.; Abdikakharovich, S.A.; Khan, A.U.; Khan, K.A.; Elboughdiri, N.; Zhu, D., 2024. Harnessing the power of bacterial laccases for xenobiotic degradation in water: a 10-year overview. Science Of The Total Environment, v. 918, 170498. https://doi.org/10.1016/j.scitotenv.2024.170498.

Reichert, G.; Hilgert, S.; Alexander, J.; Rodrigues de Azevedo, J.C.; Morck, T.; Fuchs, S.; Schwartz, T., 2021. Determination of antibiotic resistance genes in a WWTP-impacted river in surface water, sediment, and biofilm: Influence of seasonality and water quality. Science of the Total Environment, v. 768, 144526. https://doi.org/10.1016/j.scitotenv.2020.144526.

Rodrigues, F.; Durães, L.; Simões, N.E.C.; Pereira, A.M.P.T.; Silva, L.J.G.; Feio, M.J., 2025. Pharmaceuticals in urban streams: A review of their detection and effects in the ecosystem. Water Research, v. 268 (Part B), 122657. https://doi.org/10.1016/j.watres.2024.122657.

Saini, S.; Tewari, S.; Dwivedi, J.; Sharma, V., 2023. Biofilm-mediated wastewater treatment: a comprehensive review. Materials Advances, v. 4 (6), 1415-1443. https://doi.org/10.1039/d2ma00945e.

Santos, L.H.M.L.M.; Freixa, A.; Insa, S.; Acuña, V.; Sanchís, J.; Farré, M.; Sabater, S.; Barceló, D.; Rodríguez-Mozaz, S., 2019. Impact of fullerenes in the bioaccumulation and biotransformation of venlafaxine, diuron and triclosan in river biofilms. Environmental Research, v. 169, 377-386. https://doi.org/10.1016/j.envres.2018.11.036.

Shabbir, S.; Faheem, M.; Dar, A.A.; Ali, N.; Kerr, P.G.; Yu, Z.G.; Li, Y.; Frei, S.; Albasher, G.; Gilfedder, B.S., 2022. Enhanced periphyton biodegradation of endocrine disrupting hormones and microplastic: Intrinsic reaction mechanism, influential humic acid and microbial community structure elucidation. Chemosphere, v. 293, 133515. https://doi.org/10.1016/j.chemosphere.2022.133515.

Torres, N.H.; Aguiar, M.M.; Ferreira, L.F.R.; Américo, J.H.P.; Machado, Â.M.; Cavalcanti, E.B.; Tornisielo, V.L., 2015. Detection of hormones in surface and drinking water in Brazil by LC-ESI-MS/MS and ecotoxicological assessment with Daphnia magna. Environmental Monitoring and Assessment, v. 187, 379. https://doi.org/10.1007/s10661-015-4626-z.

Torres, N.H.; Santos, G.O.S.; Romanholo Ferreira, L.F.; Américo-Pinheiro, J.H.P.; Eguiluz, K.I.B.; Salazar-Banda, G.R., 2021. Environmental aspects of hormones estriol, 17β-estradiol and 17α-ethinylestradiol: Electrochemical processes as next-generation technologies for their removal in water matrices. Chemosphere, v. 267, 128888. https://doi.org/10.1016/j.chemosphere.2020.128888.

Viancelli, A.; Avalos, D.M.; Reis, P.; Málaga, P.R.S.; Shah, M.P.; Dwivedi, N.; Michelon, W., 2023. The Impact of 17β-estradiol (E2) on the Growth Profile of Environmental Enterobacteriaceae. Water, Air, & Soil Pollution, v. 234 (20). http://doi.org/10.1007/s11270-022-06036-3.

Vilela, C.L.S.; Bassin, J.P.; Peixoto, R.S., 2018. Water contamination by endocrine disruptors: Impacts, microbiological aspects and trends for environmental protection. Environmental Pollution, v. 235, 546-559. https://doi.org/10.1016/j.envpol.2017.12.098.

Wang, D.; Zhu, X.; Xi, W.; Pan, H.; Yao, H.; Du, Y.,. 2023. Purification capacity of natural biofilms and their physiochemical and biological properties: a case study in the Jishan River, a heavily polluted river. Water Supply, v. 23 (4), 1611-1625. https://doi.org/10.2166/WS.2023.078.

Yan, S.; Ding, N.; Yao, X.; Song, J.; He, W.; Rehman, F.; Guo, J., 2023. Effects of erythromycin and roxithromycin on river periphyton: Structure, functions and metabolic pathways. Chemosphere, v. 316, 137793. https://doi.org/10.1016/j.chemosphere.2023.137793.

Yuan, K.; Li, S.; Zhong, F., 2020. Treatment of coking wastewater in biofilm-based bioaugmentation process: Biofilm formation and microbial community analysis. Journal of Hazardous Materials, v. 400, 123117. https://doi.org/10.1016/j.jhazmat.2020.123117.

Zhang, C.; Li, Y.; Wang, C.; Niu, L.; Cai, W., 2016. Occurrence of endocrine disrupting compounds in aqueous environment and their bacterial degradation: A review. Critical Reviews in Environmental Science and Technology, v. 46 (1), 1-59. https://doi.org/10.1080/10643389.2015.1061881.

Zhang, F.; Yu, Y.; Pan, C.; Saleem, M.; Wu, Y., 2021. Response of periphytic biofilm in water to estrone exposure: Phenomenon and mechanism. Ecotoxicology and Environmental Safety, v. 207, 111513. https://doi.org/10.1016/j.ecoenv.2020.111513.