Environmental Science and Pollution Research

, Volume 26, Issue 18, pp 18730–18738 | Cite as

Enhanced inactivation of antibiotic-resistant bacteria isolated from secondary effluents by g-C3N4 photocatalysis

  • Ning Ding
  • Xueming Chang
  • Na Shi
  • Xiufeng Yin
  • Fei Qi
  • Yingxue SunEmail author
Research Article


The extensive use of antibiotics has resulted in the development of antibiotic-resistant bacteria (ARB), which may not be completely removed by traditional wastewater treatment processes. More effective approaches to disinfection are needed to prevent the release of ARB into the surface water. The metal-free photocatalyst graphitic carbon nitride (g-C3N4) has aroused great interest as a possible agent for water and wastewater treatment, due to its low cytotoxicity and photoactivity with visible light. In this study, the efficacy of g-C3N4 was assessed as a possible means to enhance ARB inactivation by irradiation. ARB were isolated and purified from secondary effluents in 4 municipal wastewater treatment plants. Of these, 4 typical multi-drug ARB isolates, belonging to Enterobacteriaceae, were selected for irradiation experiments. Inactivation was seen to increase with irradiation time. At 60 min, the inactivation of the 4 ARB isolates by light at > 300 nm and > 400 nm was in the range of 0.25–0.39 log and 0.16–0.19 log, respectively. The use of g-C3N4-mediated photocatalysis at the same wavelengths significantly enhanced that to 0.64–1.26 log and 0.31–0.41 log, respectively. The antibiotic susceptibility of the ARB isolates remained unchanged either prior to or after irradiation and was independent of photon fluence, reaction time, and the presence of g-C3N4. This study establishes a baseline for understanding the effectiveness of g-C3N4 photocatalysis on inactivation of ARB in wastewaters and lays the foundation for further improvement in the use of photocatalysis for wastewater treatment.


Antibiotic-resistant bacteria g-C3N4 Photocatalysis Inactivation Antibiotic susceptibility Secondary effluents 


Funding information

This work was supported by the National Natural Science Foundation of China (no. 21306003), the General Project of Beijing Educational Committee (KM201810011011), and China Scholarship Council.

Supplementary material

11356_2019_5080_MOESM1_ESM.docx (2.7 mb)
ESM 1 (DOCX 2716 kb)


  1. Allahverdiyev AM, Abamor ES, Bagirova M, Rafailovich M (2011) Antimicrobial effects of TiO2 and Ag2O nanoparticles against drug-resistant bacteria and leishmania parasites. Future Microbiol 6:933–940CrossRefGoogle Scholar
  2. Andersen SR (1993) Effects of waste water treatment on the species composition and antibiotic resistance of coliform bacteria. Curr Microbiol 26:97–103CrossRefGoogle Scholar
  3. Baier J, Maisch T, Maier M, Engel E, Landthaler M, Baumler W (2006) Singlet oxygen generation by UVA light exposure of endogenous photosensitizers. Biophys J 91:1452–1459CrossRefGoogle Scholar
  4. Besaratinia A, Kim SI, Bates SE, Pfeifer GP (2007) Riboflavin activated by ultraviolet A1 irradiation induces oxidative DNA damage-mediated mutations inhibited by vitamin C. Proc Natl Acad Sci U S A 104:5953–5958CrossRefGoogle Scholar
  5. Bouki C, Venieri D, Diamadopoulos E (2013) Detection and fate of antibiotic resistant bacteria in wastewater treatment plants: a review. Ecotoxicol Environ Saf 91:1–9CrossRefGoogle Scholar
  6. Buchanan RE, Gibbons NR (1974) Bergey’s manual of determinative bacteriology, 8th edn. Williams & Wilkins, BaltimoreGoogle Scholar
  7. CLSI (2018) Performance standards for antimicrobial susceptibility testingGoogle Scholar
  8. Devi LG, Kavitha R (2013) A review on non metal ion doped titania for the photocatalytic degradation of organic pollutants under UV/solar light: role of photogenerated charge carrier dynamics in enhancing the activity. Appl Catal B-Environ 140:559–587CrossRefGoogle Scholar
  9. Dong F, Wu L, Sun Y, Fu M, Wu Z, Lee SC (2011) Efficient synthesis of polymeric g-C3N4 layered materials as novel efficient visible light driven photocatalysts. J Mater Chem 21:15171–15174CrossRefGoogle Scholar
  10. Ferro G, Fiorentino A, Alferez MC, Polo-Lopez MI, Rizzo L, Fernandez-Ibanez P (2016) Urban wastewater disinfection for agricultural reuse: effect of solar driven AOPs in the inactivation of a multidrug resistant E-coli strain. Appl Catal B-Environ 178:65–73CrossRefGoogle Scholar
  11. Flach C-F, Genheden M, Fick J, Larsson DGJ (2018) A comprehensive screening of Escherichia coli isolates from Scandinavia’s largest sewage treatment plant indicates no selection for antibiotic resistance. Environ Sci Technol 52:11419–11428CrossRefGoogle Scholar
  12. Gomez-Pastora J, Dominguez S, Bringas E, Rivero MJ, Ortiz I, Dionysiou DD (2017) Review and perspectives on the use of magnetic nanophotocatalysts (MNPCs) in water treatment. Chem Eng J 310:407–427CrossRefGoogle Scholar
  13. Guan Y, Wang B, Gao Y, Liu W, Zhao X, Huang X, Yu J (2018) Occurrence and fate of antibiotics in the aqueous environment and their removal by constructed wetlands in China: a review. Pedosphere 27:42–51CrossRefGoogle Scholar
  14. Hernandez F, Calisto-Ulloa N, Gomez-Fuentes C et al (2019) Occurrence of antibiotics and bacterial resistance in wastewater and sea water from the Antarctic. J Hazard Mater 363:447–456CrossRefGoogle Scholar
  15. Huang J, Ho W, Wang X (2014) Metal-free disinfection effects induced by graphitic carbon nitride polymers under visible light illumination. Chem Commun 50:4338–4340CrossRefGoogle Scholar
  16. Hwangbo M, Claycomb E, Liu Y, Alivio T, Alivio T, Banerjee S, Chu K (2019) Effectiveness of zinc oxide-assisted photocatalysis for concerned constituents in reclaimed wastewater: 1,4-dioxane, trihalomethanes, antibiotics, antibiotic resistant bacteria (ARB), and antibiotic resistance genes (ARGs). Sci Total Environ 649:1189–1197CrossRefGoogle Scholar
  17. Ito K, Hiraku Y, Kawanishi S (2007) Photosensitized DNA damage induced by NADH: site specificity and mechanism. Free Radic Res 41:461–468CrossRefGoogle Scholar
  18. Kang S, Huang W, Zhang L, He M, Xu S, Sun D, Jiang X (2018a) Moderate bacterial etching allows scalable and clean delamination of g-C3N4 with enriched unpaired electrons for highly improved photocatalytic water disinfection. ACS Appl Mater Interfaces 10:13796–13804CrossRefGoogle Scholar
  19. Kang S, Zhang L, He M, Zheng Y, Cui L, Sun D, Hu B (2018b) “Alternated cooling and heating” strategy enables rapid fabrication of highly-crystalline g-C3N4 nanosheets for efficient photocatalytic water purification under visible light irradiation. Carbon 137:19–30CrossRefGoogle Scholar
  20. Karaolia P, Michael I, Garcia-Fernandez I, Aguera A, Malato S, Fernandez-Ibanez P, Fatta-Kassinos D (2014) Reduction of clarithromycin and sulfamethoxazole-resistant Enterococcus by pilot-scale solar-driven Fenton oxidation. Sci Total Environ 468:19–27CrossRefGoogle Scholar
  21. Li Y, Zhang C, Shuai D, Naraginti S, Wang D, Zhang W (2016) Visible-light-driven photocatalytic inactivation of MS2 by metal-free g-C3N4: virucidal performance and mechanism. Water Res 106:249–258CrossRefGoogle Scholar
  22. Lin LS, Cong ZX, Li J, Ke KM, Guo SS, Yang HH, Chen GN (2014) Graphitic-phase C3N4 nanosheets as efficient photosensitizers and pH-responsive drug nanocarriers for cancer imaging and therapy. J Mater Chem B 2:1031–1037CrossRefGoogle Scholar
  23. Lubart R, Lipovski A, Nitzan Y, Friedmann H (2010) A possible mechanism for the bactericidal effect of visible light. Laser Ther 20:17–22CrossRefGoogle Scholar
  24. Maeda K, Wang X, Nishihara Y, Lu D, Antonietti M, Domen K (2009) Photocatalytic activities of graphitic carbon nitride powder for water reduction and oxidation under visible light. J Phys Chem C 113:4940–4947CrossRefGoogle Scholar
  25. Makowska N, Koczura R, Mokracka J (2016) Class 1 integrase, sulfonamide and tetracycline resistance genes in wastewater treatment plant and surface water. Chemosphere 144:1665–1673CrossRefGoogle Scholar
  26. Mao D et al (2015) Prevalence and proliferation of antibiotic resistance genes in two municipal wastewater treatment plants. Water Res 85:458–466CrossRefGoogle Scholar
  27. Martins da Costa P, Vaz-Pires P, Bernardo F (2006) Antimicrobial resistance in Enterococcus spp. isolated in inflow, effluent and sludge from municipal sewage water treatment plants. Water Res 40:1735–1740CrossRefGoogle Scholar
  28. Matsunaga T, Tomoda R, Nakajima T, Wake H (1985) Photoelectrochemical sterilization of microbial cells by semiconductor powders. FEMS Microbiol Lett 29:211–214CrossRefGoogle Scholar
  29. McConnell MM, Hansen LT, Jamieson RC, Neudorf KD, Yost CK, Tong A (2018) Removal of antibiotic resistance genes in two tertiary level municipal wastewater treatment plants. Sci Total Environ 643:292–300CrossRefGoogle Scholar
  30. Michael I, Hapeshi E, Michael C, Varela AR, Kyriakou S, Manaia CM, Fatta-Kassinos D (2012) Solar photo-Fenton process on the abatement of antibiotics at a pilot scale: degradation kinetics, ecotoxicity and phytotoxicity assessment and removal of antibiotic resistant enterococci. Water Res 46:5621–5634CrossRefGoogle Scholar
  31. Michael-Kordatou I, Karaolia P, Fatta-Kassinos D (2018) The role of operating parameters and oxidative damage mechanisms of advanced chemical oxidation processes in the combat against antibiotic-resistant bacteria and resistance genes present in urban wastewater. Water Res 129:208–230CrossRefGoogle Scholar
  32. Pan M, Chu LM (2018) Occurrence of antibiotics and antibiotic resistance genes in soils from wastewater irrigation areas in the Pearl River Delta region, southern China. Sci Total Environ 624:145–152CrossRefGoogle Scholar
  33. Pawlowski AC, Wang W, Koteva K, Barton HA, McArthur AG, Wright GD (2016) A diverse intrinsic antibiotic resistome from a cave bacterium. Nat Commun 7Google Scholar
  34. Peng S, Shuyu J, Xu-Xiang Z, Tong Z, Shupei C, Aimin L (2013) Metagenomic insights into chlorination effects on microbial antibiotic resistance in drinking water. Water Res 47:111–120CrossRefGoogle Scholar
  35. Pruden A, Arabi M, Storteboom H (2012) Correlation between upstream human activities and riverine antibiotic resistance genes. Environ Sci Technol 46:11541–11549CrossRefGoogle Scholar
  36. Pugazhenthiran N, Murugesan S, Anandan S (2013) High surface area Ag-TiO2 nanotubes for solar/visible-light photocatalytic degradation of ceftiofur sodium. J Hazard Mater 263:541–549CrossRefGoogle Scholar
  37. Rizzo L, Sannino D, Vaiano V, Sacco O, Scarpa A, Pietrogiacomi D (2014) Effect of solar simulated N-doped TiO2 photocatalysis on the inactivation and antibiotic resistance of an E. coli strain in biologically treated urban wastewater. Appl Catal B-Environ 144:369–378CrossRefGoogle Scholar
  38. Sinha RP, Hader DP (2002) UV-induced DNA damage and repair: a review. Photochem Photobiol Sci 1:225–236CrossRefGoogle Scholar
  39. Tsai TM, Chang HH, Chang KC, Liu YL, Tseng CC (2010) A comparative study of the bactericidal effect of photocatalytic oxidation by TiO2 on antibiotic-resistant and antibiotic-sensitive bacteria. J Chem Technol Biotechnol 85:1642–1653CrossRefGoogle Scholar
  40. Wang X, Maeda K, Thomas A et al (2009) A metal-free polymeric photocatalyst for hydrogen production from water under visible light. Nat Mater 8:76–80CrossRefGoogle Scholar
  41. Wu T, Liu G, Zhao J, Hidaka H, Serpone N (1998) Photoassisted degradation of dye pollutants. V. Self-photosensitized oxidative transformation of Rhodamine B under visible light irradiation in aqueous TiO2 dispersions. J Phys Chem B 102:5845–5851CrossRefGoogle Scholar
  42. Xu SJ, Shen JQ, Chen S, Zhang MH, Shen T (2002) Active oxygen species (1O2, O2·) generation in the system of TiO2 colloid sensitized by hypocrellin B. J Photochem Photobiol B-Biol 67:64–70CrossRefGoogle Scholar
  43. Xu J, Xu Y, Wang H et al (2015) Occurrence of antibiotics and antibiotic resistance genes in a sewage treatment plant and its effluent-receiving river. Chemosphere 119:1379–1385CrossRefGoogle Scholar
  44. Xu J, Wang Z, Zhu Y (2017) Enhanced visible-light-driven photocatalytic disinfection performance and organic pollutant degradation activity of porous g-C3N4 nanosheets. Acs Appl Mater Inter 9:27727–27735CrossRefGoogle Scholar
  45. Yan SC, Li ZS, Zou ZG (2009) Photodegradation performance of g-C3N4 fabricated by directly heating melamine. Langmuir 25:10397–10401CrossRefGoogle Scholar
  46. Zhang T, Wang X, Zhang X (2014) Recent progress in TiO2-mediated solar photocatalysis for industrial wastewater treatment. Int J Photoenergy:607954Google Scholar
  47. Zhang Q-Q, Ying G-G, Pan C-G, Liu Y-S, Zhao J-L (2015) Comprehensive evaluation of antibiotics emission and fate in the river basins of China: source analysis, multimedia modeling, and linkage to bacterial resistance. Environ Sci Technol 49:6772–6782CrossRefGoogle Scholar
  48. Zhang C, Li Y, Shuai DM, Zhang WL, Niu LH, Wang LF, Zhan HJ (2018a) Visible-light-driven, water-surface-floating antimicrobials developed it from graphitic carbon nitride and expanded perlite for water disinfection. Chemosphere 208:84–92CrossRefGoogle Scholar
  49. Zhang C, Li Y, Zhang W, Wang P, Wang C (2018b) Metal-free virucidal effects induced by g-C3N4 under visible light irradiation: statistical analysis and parameter optimization. Chemosphere 195:551–558CrossRefGoogle Scholar
  50. Zhang C, Li Y, Shuai D, Shen Y, Xiong W, Wang L (2019) Graphitic carbon nitride (g-C3N4)-based photocatalysts for water disinfection and microbial control: a review. Chemosphere 214:462–479CrossRefGoogle Scholar
  51. Zheng Q, Durkin D, Elenewski J et al (2016) Visible-light-responsive graphitic carbon nitride: rational design and photocatalytic applications for water treatment. Environ Sci Technol 50:12938–12948CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Ning Ding
    • 1
    • 2
  • Xueming Chang
    • 1
  • Na Shi
    • 3
  • Xiufeng Yin
    • 1
  • Fei Qi
    • 1
  • Yingxue Sun
    • 1
    • 2
    Email author
  1. 1.Department of Environmental Science and EngineeringBeijing Technology and Business UniversityBeijingChina
  2. 2.Key Laboratory of Cleaner Production and Comprehensive Utilization of Resources, China National Light IndustryBeijing Technology and Business UniversityBeijingChina
  3. 3.Beijing Boda Water Co., LtdBeijingChina

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