Screening of Antibiotics Biodegradability from Wastewater

Year 2023, , 70 - 85, 26.06.2023

Souhila Saım , Mokrani Slimane Isabel Martínez-alcalá Ramazan Erenler

https://doi.org/10.26650/EurJBiol.2023.1299300

Abstract

Objective: One of the sources of environment antibiotics contamination is wastewater treatment plants (WWTPs), there by constituting a global public health risk. This present study aimed to investigate the biodegradability of antibiotics and antiseptics and highlights the biodegradation of Ciprofloxacin as a sole carbon source by a bacterium isolated from the sludge "El Kouwaer," WWTP located in Mascara.
Materials and Methods: In the present study, biodegradability of some antibiotics and antiseptic were tested at 50 mg/l concentration through active sludge microorganisms by Manometric Respirometry Method (OECD 301F). Further analysis of 16S rRNA gene sequencing used to identify MK4 strain isolated from the sludge. Furthermore, ATR-FTIR spectroscopy was conducted in order to identify its biodegradation in the presence of different carbon sources and LCMS/MS spectrometry were used to identify the metabolite degradation.
Results: Our Results revealed that four antibiotics tested were readily biodegradable (60%) as Ciprofloxacin, Doxycycline, Amoxicillin, Ampicillin, and Penicillin. Conversely, other was not readily biodegradable, such as Azithromycin (36.11%), Cephalexin (36.20%), and Metronidazole (33.33%). Meanwhile, the remaining antibiotics under examination were degraded, with Sulfamethoxazole (25.75%), Clarithromycin (25.36%), and Nifuroxazide (16.33%) recording degradation. Ciprofloxacin was chosen to represent the most biodegraded antibiotic. Based on 16S rRNA gene, MK4 strain was related to Klebsiella oxytoca (99.99%). ATR-FTIR revealed that the strain K. oxytoca MK4 caused changes in the structure of the Ciprofloxacin, in the presence of various sources of carbon, with varying effects on bacterial growth and biodegradation.
Conclusion: In this study, the identified strain K. oxytoca MK4 facilitated the degradation of Ciprofloxacin.

Keywords

Wastewater, antibiotic, antiseptics, biodegradability, ATR-FTIR spectroscopy, LCMS/MS metabolites.

References

• 1. Yang YSO, Kim KH, Kwon EE, Tsang YF. Occurrences and removal of pharmaceuticals and personal care products (PPCPs) in drinking water and water/sewage treatment plants: A review. Sci Total Environ. 2017;596-597:303-320. google scholar
• 2. Liu H, Pu C, Yu X, Sun Y, Chen J. Removal of tetracyclines, sulfonamides, and quinolones by industrial-scale composting and anaerobic digestion processes. Environ Sci Pollut and Environ Monitoring. 2018;18:313-318. google scholar
• 3. Amorim IS, Moreira AS, Maia ME, Tiritan PM, Castro P. Biodegradation of ofloxacin, norfloxacin, and ciprofloxacin as single and mixed substrates by Labrys portucalensis F11. Appl Microbiol Biotechnol. 2014;98:3181-3190. google scholar
• 4. Kumar A, Pal D. Antibiotic resistance and wastewater: Correla-tion, impact and critical human health challenges. J Environ Chem Eng. 2018;6: 52-58. google scholar
• 5. Carvalho RN, Ceriani L, Ippolito A, Lettieri T. Development of the first Watch List under the Environmental Quality Standards Directive. EUR-Scientific and Technical Research series 2015 -ISSN 1831-9424-166. google scholar
• 6. Sabri NA, van Holst S, Schmitt H, et al., Fate of antibiotics and antibiotic resistant genes during conventional and addi-tional treatment technologies in wastewater treatment plants. Sci Total Environ. 2020;741:140199. google scholar
• 7. Sorinolu AJ, Tyagi N, Kumar A, Mariya Munir M. Antibiotic resistance development and human health risks during wastewa-ter reuse and biosolids application in agriculture. Chemosphere. 2021; 265:129032. google scholar
• 8. Aziz A, Sengar A, Basheer F, Farooqi IH, Isa MH. Anaerobic digestion in the elimination of antibiotics and antibiotic-resistant genes from the environment-A comprehensive review. J Environ Chem Eng. 2022;10(1):106423. google scholar
• 9. Kinigopoulou V, Ioannis P, Kalderis D, Anastopoulos I. Mi-croplastics as carriers of inorganic and organic contaminants in the environment: A review of recent progress. J Mol Liq. 2022; 350:118580. google scholar
• 10. Sahlin, S, Larsson JD. Agerstrand M.CIP: EQS Data Overview. In The Department of Environmental Science and Analytical Chem-istry (ACES) Report; Stockholms Universitet: Stockholm, Swe-den, 2018; p.15. google scholar
• 11. Dorival-Garcia N, Zafra-Gomez A, Navalon A, Gonzalez J, Vilchez JL. Removal of quinolone antibiotics from wastewaters by sorption and biological degradation in laboratory-scale mem-brane bioreactors. Sci Total Environ. 2013; 442:317-328. google scholar
• 12. Liu Z, Sun P, Pavlostathis SG, Zhou X, Zhang Y. Adsorption, inhibition, and biotransformation of ciprofloxacin under aerobic conditions. Bioresour Technol. 2013;144: 644-651 google scholar
• 13. Alexandrino DA, Mucha AP, Almeida CMR, Gao W, Jia Z, Carvalho MF. Biodegradation of the veterinary antibiotics en-rofloxacin and ceftiofur and associated microbial community dy-namics. Sci Total Environ. 2017;581:3593. google scholar
• 14. Lima VB, Goulart LA, Rocha RS, Steter JR, Lanza MR. Degra-dation of antibiotic ciprofloxacin by different AOP systems using electrochemically generated hydrogen peroxide. Chemosphere. 2020;125807. google scholar
• 15. Wang J, Mao D, Mu Q, Luo Y. Fate and proliferation of typi-cal antibiotic resistance genes in five full-scale pharmaceutical wastewater treatment plants. Sci Total Environ. 2015;526:366-373. google scholar
• 16. OECD 310. Ready biodegradability-CO2 in sealed vessels (Headspace test). 2014 google scholar
• 17. Osinska A, Korzeniewska E, Harnisz M, Niestçpski S. The preva-lence and characterization of antibiotic-resistant and virulent Escherichia coli strains in the municipal wastewater system and their environmental fate. Sci Total Environ. 2017; 577: 367-375. google scholar
• 18. OECD. Guideline for Testing of Chemicals (301 D). Closed Bottle Test. Organisation of Economic Cooperationand Development, Paris. 1992. google scholar
• 19. Arrete interministeriel fixant la liste des cultures pouvant etre imguees avec des eaux usees epurees du 2 janvier 2012. Journal Officiel de la Republique Algerieme (JORADP).2012;41:21. google scholar
• 20. American Public Health Association, Water Pollution Control Federation, Water Environment Federation. Standard methods for the examination of water and wastewater. Washington, D.C., USA: American Public Health Association; 2005. google scholar
• 21. Brown DM, Hughes CB, Spence M, Bonte M, Whale G. As-sessing the suitability of a manometric test system for determin-ing the biodegradability of volatile hydrocarbons. Chemosphere. 2018;195:381-389. google scholar
• 22. Liyanage GY, Pathmalal MM. Risk of prophylactic antibiotics in livestock and poultry farms; a growing problem for human and animal health and for the environment. Proceeding of 2nd Environment and Natural Resources International Conference; 20. 2016; Thailand. google scholar
• 23. Liyanage GY, Pathmalal MM. Isolation and characterization of oil degrading bacteria from coastal waters and sediments in Sri Lanka. J Natn Sci Foundation. 2016; 44(4):351-358. google scholar
• 24. Heuer H, Krsek M, Baker P, Smalla K, Wellington E. Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in dena-turing gradients. Appl Environ Microbiol.1997;63:3233-3241. google scholar
• 25. NCBI (2023) Available from: http://www.ncbi.nlm.nih.gov/. Ac-cessed on 22 March 2023. google scholar
• 26. Tamura K, Stecher G, Kumar S. MEGA 11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol. 2021;38(7):3022-3027. google scholar
• 27. Saitou N, Nei M. The neighbor-joining method. A new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987;4:406-425. google scholar
• 28. Felsenstein J. Confidence limits on phylogenies: An approach using the bootstrap. Evolution. 1985; 39:783-791. google scholar
• 29. Tamura K, Nei M, Kumar S. Prospects for inferring very large phy-logenies by using the neighbor-joining method. Proc Natl Acad Sci (USA). 2004;101:11030-11035. google scholar
• 30. Lan-jia P, Bao-wei Y, Jun L, Xiao-li S, Jian-Hua S. Effect of sodium acetate on ciprofloxacin elimination and micro-bial community in a sequencing batch reactor. Chemosphere. 2017;168:1479-1485. google scholar
• 31. Wackerlig J, Schirhagl R. Biodegradation of UV absorbers in wastewater using moving bed biofilm reactors. Water Res. 2015; 69:1-10. google scholar
• 32. Pan LJ, Li J, Li CX, Yu GW, Wang Y. Study of ciprofloxacin biodegradation by a Thermus sp. isolated from pharmaceutical sludge. J Hazard Mater. 2018;343:59-67. google scholar
• 33. Garland JL, Mills AL. Classification and characterization of heterotrophic microbial communities on the basis of patterns of community level sole-carbon-source utilization. Appl Environ Microbiol. 1991; 57(8):2351-2359. google scholar
• 34. Touyer M, Lashermes G, Baudez, JC. Effect of temperature on the kinetics of organic matter degradation in anaerobic sludge digesters. Bioresour Technol. 1998;63(2): 141-147. google scholar
• 35. FAO, Irrigation with treated wastewater: Operating manual, re-gional office for the near East and North Africa, Cairo, Egypt, 2003. google scholar
• 36. Painter HA. Detailed review paper on biodegradability testing (No. 98). Organisation for Economic Co-Operation and Develop-ment. 1995. google scholar
• 37. Nouha K, Yan S, Tyagi RD, Surampalli RY. EPS producing mi-croorganisms from municipal wastewater activated sludge. Civil Engineering, 2016. google scholar
• 38. Xu S, Yao J, Ainiwaer M, Hong Y, Zhang Y. Analysis of bac-terial community structure of activated sludge from wastewater treatment plants in winter. Biomed Res Int. 2018;2018:8278970. doi:10.1155/2018/8278970. google scholar
• 39. Sukumaran M, Rama Murthy V, Raveendran S, et al. Biodiversity of Microbes in Tannery Effluent. J Ecotoxicol. 2008. google scholar
• 40. Bechac JP, Boutin P, Mercier B, Nuer P. Traitements des eaux usees; Eyrolles: Paris, France, 1987. google scholar
• 41. Abdouni AE, Bouhout S, Merimi I, Hammouti B, Haboubi K. Physicochemical characterization of wastewater from the Al-Hoceima slaughterhouse in Morocco. Caspian J Environ Sci. 2021; 19(3):423-429. google scholar
• 42. Bliefert C, Perraud, D. Mass balances of organic matter and nu-trients in the biodegradation of hydrocarbons. J Hazard Mater. 2009; 163(2-3): 1185-1192. google scholar
• 43. Kümmerer K, Al-Ahmad A, Mersch-Sundermann V. Biodegrad-ability of some antibiotics, elimination of the genotoxicity and affection of wastewater bacteria in a simple test. Chemosphere. 2000; 40(7):701-710. google scholar
• 44. Seyfried M, Boschung A, Miffon F, Ohleyer E, Chaintreau A. Elu-idation of the upper pathway of alicyclic musk Romandolide® degradation in OECD screening tests with activated sludge. Environ Sci Pollut Res. 2014;21(16):9487-9494. google scholar
• 45. ECHA. Guidance on information requirements and chemical safety assessment 2017; chapter R.11: PBT/vPvB Assessment. Version 4.0, June 2017. google scholar
• 46. O’Malley LP. Evaluation and modification of the OECD 301F Respirometry biodegradation test method with regards to test sub-stance concentration and inoculum. Water Air Soil Pollut. 2006; 177(1-4):251-265. google scholar
• 47. Guzialowska-Tic K, Tic W. Biodegradation of selected volatile organic compounds in groundwater. J Ecol Engineering. 2016; 17(3):145-150. google scholar
• 48. Bergheim M, Giere R, Kümmerer K. Biodegradability and eco-toxicity of tramadol, ranitidine, and their photoderivatives in the aquatic environment. Environ Sci Pollut Res. 2012;19(1):72-85. google scholar
• 49. Piçtka-Ottlik M, Fr^ckowiak R, Maliszewska I, Kolwzan B, Wilk KA. Ecotoxicity and biodegradability of antielectrostatic dicephalic cationic surfactants. Chemosphere. 2012; 89(9):1103-1111. google scholar
• 50. Cristobal-Garrfa J, Gonzâlez-Martmez A, Dionysiou DD Biodegradation of pharmaceutical compounds: A review. J Hazard Mater. 2011;167(1-3):355-367. google scholar
• 51. Al-Ahmad A, Daschner FD, Kümmerer K. Biodegradabil-ity of Cefotiam, ciprofloxacin, Meropenem, Penicillin G, and Sulfamethoxazole and inhibition of wastewater bacteria. Arch Environ Contam Toxicol. 1999;37:158-163. google scholar
• 52. Deng Y, Zhang Y, Gao Y, et al. Microbial community compo-sitional analysis for series reactors treating high level antibiotic wastewater. Environ Sci Technol. 2012; 46:795-801. google scholar
• 53. Li L, Guo C, Fan S, et al. Dynamic transport of antibiotics and antibiotic resistance genes under different treatment processes in a typical pharmaceutical wastewater treatment plant. Environ Sci Pollut Res. 2018; 25:30191-30198. google scholar
• 54. Blackwell PA, Kay P, Pemberton EJ, Croxford A. Veterinary medicines in the environment. Rev Environ Contam Toxicol. 2007; 180:1-91. google scholar
• 55. Li H, Helm PA, Metcalfe CD. Sampling in the Great Lakes for pharmaceuticals, personal care products, and endocrine-disrupting substances using the passive polar or-ganic chemicalintegrative sampler. Environ Toxicol Chem. 2010; 29(4):751-762.https://doi.org/10.1002/ETC. google scholar
• 56. Bertelkamp C, Reungoat J, Cornelissen ER, et al. Sorption and biodegradation of organic micropollutants during river bank fil-tration: A laboratory column study. Water Res. 2014; 52:231-241. doi:10.1016/j.watres.2013.10.068 google scholar
• 57. Cetecioglu Z, Bahar I, Meritxell G, Sara Rodriguez-Mozaz, Damia B, Orhan I, Derin O. Biodegradation and reversible in-hibitory impact of sulfamethoxazole on the utilization of volatile fatty acids during anaerobic treatment of pharmaceutical industry wastewater. Sci Total Environ. 2015; 536:667-674. google scholar
• 58. Prado B, Seco A, Ferrero L, Fdz-Polanco M. Behaviour of tetra-cyclines in soils: Mobility, degradation, and interaction with clay fractions. J Environ Sci Health. Part B. 2009;44(1):72-83. google scholar
• 59. Gartiser S, Urich E, Alexy R, Kümmerer K. Ultimate biodegrada-tion and elimination of antibiotics in inherent tests. Chemosphere. 2007;67:604-613. doi: 10.1016/j.chemosphere.2006.08.038 google scholar
• 60. Nyholm N. The European system of standardised legal tests for as-sessing the biodegradability of chemicals. Environ Toxicol Chem. 1991;11:1237-1246. google scholar
• 61. Pepper IL, Charles PG, Terry J. G. Environmental microbiology. Third edition. United States of America. 2015. google scholar
• 62. Luo Y, Xu L, Rysz M, Wang Y, Zhang H, Alvarez PJ. Occur-rence and transport of tetracycline, sulfonamide, quinolone, and macrolide antibiotics in the Haihe River Basin, China. Environ Sci Technol. 2011; 45(5):1827-1833. google scholar
• 63. Chen Y, Zhiping W, Lili L, Hanbin Z, Pin W. Stress-responses of microbial population and activity in activated sludge un-der long-term ciprofloxacin exposure. J Environ Manage. 2021; 281:111896. google scholar
• 64. Decre D, Burghoffer B, Gautier V, Petit JC, Arlet G. Out-break of multi-resistant Klebsiella oxytoca involving strains with extended-spectrum beta-lactamases and strains with extended-spectrum activity of the chromosomal beta-lactamase. J Antimicrob Chemother. 2004;54:881-888. google scholar
• 65. Lowe C, Willey B, O’Shaughnessy A, et al., Mount Sinai Hos-pital Infection Control Team. Outbreak of extended-spectrum beta-lactamase-producing Klebsiella oxytoca infections associ-ated with contaminated handwashing sinks (1). Emerg Infect Dis. 2012; 18:1242-1247. google scholar
• 66. Schulz-Stubner S, Kniehl E. Transmission of extended-spectrum beta-lactamase Klebsiella oxytoca via the breathing circuit of a transport ventilator: root cause analysis and infec-tion control recommendations. Infect Control Hosp Epidemiol. 2011;32:828-829. google scholar
• 67. Zârate MS, Gales AC, Picâo RC, Pujol GS, Lanza A, Smayevsky J. Outbreak of OXY-2-producing Klebsiella oxytoca in a renal transplant unit. J Clin Microbiol. 2008; 46:2099-2101. google scholar
• 68. Gheorghiu R, Yuan M, Hall LM, Livermore DM. Bases of varia-tion in resistance to beta-lactams in Klebsiella oxytoca isolates hyperproducing K1 beta-lactamase. J Antimicrob Chemother. 1997;40:533-541. google scholar
• 69. Paterson DL, Bonomo RA. Extended-spectrum beta-lactamases: a clinical update. Clin Microbiol Rev. 2005; 18:657-686. google scholar
• 70. Brisse S, Verhoef J. Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes se-quencing and automated ribotyping. Int J Syst Evol Microbiol. 2001;51:915-924. google scholar
• 71. Wang Y, Zhang S, Ye J, Huang J, Jiang X. Enhanced degrada-tion of norfloxacin and sulfamethoxazole in aqueous solution by Klebsiella oxytoca under aerobic conditions. Ecotoxicol Environ Saf. 2020; 201:110843. doi: 10.1016/j.ecoenv.2020.110843 google scholar
• 72. Xiong MB, Kurade JR, Kim HS, Roh B, Jeon H. Ciprofloxacin toxicity and its co-metabolic removal by a freshwater microalga Chlamydomonas mexicana. J Hazard Mater. 2016; 323:212-219. google scholar
• 73. Pan LJ, Tang XD, Li CX, Yu GW, Wang Y. Biodegrada-tion of sulfamethazine by an isolated Thermophile-Geobacillus sp. S-07. World J Microbiol Biotechnol. 2017;33(5):85. doi:10.1007/s11274-017-2245-2 google scholar
• 74. Guo HG, Gao NY, Chu WH, et al., Photochemical degradation of ciprofloxacin in UV and UV/H2O2 process: kinetics, parameters, and products. Environ Sci Poll Research. 2013; 20(5) :3202- 3213. google scholar
• 75. Tan Z, Tan F, Zhao L, Li J. The Synthesis, characterization and application of ciprofloxacin complexes and its coordination with Copper, Manganese and Zirconium Ions. J Cryst Process Technol. 2012;2(02) :55 google scholar
• 76. Yan SW, Chen HY, Xiao DR, et al. An unprecedented 2D—3D polythreaded metal-lomefloxacin complex assembled from sidearm-containing 2D motifs. Inorg Chem Commun. 2012;15: 47-51. DOI: 10.1016/j.inoche.2011.09.036. google scholar
• 77. Prieto A, Möder M, Rodil R, Adrian L, Marco-Urrea E. Degrada-tion of the antibiotics norfloxacin and ciprofloxacin by a white-rot fungus and identification of degradation products. Biores Technol. 2011;102(23):10987-10995. google scholar
• 78. Parshikov I, Heinze T, Moody J, Freeman J, Williams A, Suther-land J. The funguPestalotiopsis guepini as a model for bio-transformation of ciprofloxacin and norfloxacin. App Microbiol Biotechnol. 2001; 56(3-4):474-477. google scholar
• 79. Dewitte B, Dewulf J, Demeestere K, et al., Ozonation of ciprofloxacin in water: HRMS identification of reaction products and pathways. Environ Sci Technol. 2008; 42(13): 4889-4895. google scholar
• 80. Santoke H, Song W, Cooper WJ, Greaves J, Miller GE. Free-radicalinduced oxidative and reductive degradation of fluoro-quinolone pharmaceuticals: kinetic studies and degradation mech-anism. J Physical Chem A. 2009;113(27):7846-7851. google scholar
• 81. Nguyen TT, Bui XT, Dang BT, et al., Effect of ciprofloxacin dosages on the performance of sponge membrane bioreactor treat-ing hospital wastewater. Bioresour Technol. 2019; 273: 573-580. google scholar
• 82. Polesel F, Lehnberg K, Dott W, Trapp S, Thomas KV, Plosz BG. Factors influencing sorption of ciprofloxacin onto activated sludge: Experimental assessment and modelling implications. Chemosphere. 2015;119:105-111. google scholar
• 83. Shokoohi R, Ghobadi N, Godini K, Hadi M, Atashzaban Z. An-tibiotic detection in a hospital wastewater and comparison of their removal rate by activated sludge and earthworm-based vermifilter-ation: Environmental risk assessment. Process Saf Environ Prot. 2020;134:169-177. google scholar
• 84. Xiaobin L, Bingxin L, Rusen Z,Yu D, Shuguang X, Baoling Y. Biodegradation of antibiotic ciprofloxacin: Pathways, influential factors, and bacterial community structure. Environ Sci Pollut Res. 2016;23:7911-7918. google scholar
• 85. Feng NX, Yu J, Xiang L, et al. Co-metabolic degradation of the antibiotic ciprofloxacin by the enriched bacterial consortium XG and its bacterial community composition. Sci Total Environ. 2019;665:41-51. google scholar
• 86. Wetzstein HG, Stadler M, Tichy HV, Dalhoff A, Karl W. Degra-dation of 650 ciprofloxacin by basidiomycetes and identifica-tion of metabolites generated by the brown 651 rot fungus Gloeophyllum striatum. Appl Environ Microb. 1999;65(4):1556-1563. google scholar
• 87. Wetzstein HG, Schneider J, Karl W. Patterns of metabolites pro-duced from the fluoroquinolone norfloxacin by basidiomycetes indigenous to agricultural sites. Appl Microbiol Biotechnol. 2006; 71: 90-100. google scholar
• 88. Cvancarova M, Moeder M, Filipova A, Cajthaml T. Bio-transformation of fluoroquinolone antibiotics by ligninolytic fungi-metabolites, enzymes and residual antibacterial activity. Chemosphere.2015; 136:311-320. google scholar
• 89. Jia Y, Khanal SK, Shu H, Zhang H, Chen GH, LuH. Ciprofloxacin degradation in anaerobic sulfate-reducing bacteria (SRB) sludge system: Mechanism and pathways. Water Res. 2018;136:64-74. google scholar
• 90. Jung CM, Heinze TM, Strakosha R, Elkins CA, Sutherland JB. Acetylation of fluoroquinolone antimicrobial agents by an Escherichia coli strain isolated from a municipal wastewater treat-ment plant. J Appl Microbiol. 2009; 106(2):564-571. google scholar