Abstract
The occurrence of clarithromycin in wastewater samples and of the activated sludge bacteria possibly resistant to this pharmaceutical was the object of the study. Samples of wastewater or activated sludge were taken from a municipal wastewater treatment plant in summer and winter and characterised regarding their clarithromycin concentrations and the presence of nucleic acid fragments (Cla-sequences) known to be responsible for clarithromycin resistance in Helicobacter pylori. The concentrations of clarithromycin in raw wastewater were about 1086–2271 ng/L. Around 50–60% less of the pharmaceutical was found in treated wastewater. The concentrations were much higher in winter samples, as compared to summer samples. The clarithromycin resistance markers in H. pylori were detected by fluorescence in situ hybridisation in activated sludge bacterial cells. Cla-sequences were found in all the detected Proteobacteria, independently of the sampling season. Among nitrifying or phosphate or glycogen accumulating bacteria only Nitrosomonas spp. revealed presence of the clarithromycin sequences.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahn Y, Choi J (2016) Bacterial communities and antibiotic resistance communities in a full-scale hospital wastewater treatment plant by high-throughput pyrosequencing. Water 8:580. https://doi.org/10.3390/w8120580
Alfaresi MS, Elkoush AA (2010) Characterization of clarithromycin resistance in isolates of Helicobacter pylori from the UAE. Indian J Gastroenterol 29:116–120. https://doi.org/10.1007/s12664-010-0034-z
Bai X, Xi C, Wu J (2016) Survival of Helicobacter pylori in the wastewater treatment process and the receiving river in Michigan, USA. J Water Health 14:692–698. https://doi.org/10.2166/wh.2016.259
Barancheshme F, Munir M (2018) Strategies to combat antibiotic resistance in the wastewater treatment plants. Front Microbiol. https://doi.org/10.3389/fmicb.2017.02603
Bellanger X, Guilloteau H, Bonot S, Merlin C (2014) Demonstrating plasmid-based horizontal gene transfer in complex environmental matrices: a practical approach for a critical review. Sci Total Environ 493:872–882. https://doi.org/10.1016/j.scitotenv.2014.06.070
Bouki C, Venieri D, Diamadopoulos E (2013) Detection and fate of antibiotic resistant bacteria in wastewater treatment plants: a review. Ecotoxicol Environ Saf 91:1–9. https://doi.org/10.1016/j.ecoenv.2013.01.016
Coenen S, Ferech M, Malhotra-Kumar S, Hendrickx E, Suetens C, Goossens H (2006) European Surveillance of Antimicrobial Consumption (ESAC): outpatient macrolide, lincosamide and streptogramin (MLS) use in Europe. J Antimicrob Chemother 58:418–422. https://doi.org/10.1093/jac/dkl184
Costanzo SD, Murby J, Bates J (2005) Ecosystem response to antibiotics entering the aquatic environment. Mar Pollut Bull 51:218–223. https://doi.org/10.1016/j.marpolbul.2004.10.038
Daims H, Stoecker K, Wagner M (2005) Fluorescence in situ hybridization for the detection of procaryotes. In: Osborn AM, Smith CJE (eds) Advanced methods in molecular microbial ecology. Bios-Garland, Abingdon
European Centre for Disease Prevention and Control (ECDPC) (2018) Antimicrobial consumption. In: ECDC. Annual epidemiological report 2017. ECDC, Stockholm
Ferreira Da Silva M, Vaz-Moreira I, Gonzalez-Pajuelo M, Nunes OC, Manaia CM (2007) Antimicrobial resistance patterns in Enterobacteriaceae isolated from an urban wastewater treatment plant. FEMS Microbiol Ecol 60:166–176. https://doi.org/10.1111/j.1574-6941.2006.00268.x
Forster S, Snape JR, Lappin-Scott HM, Porter J (2002) Simultaneous fluorescent gram staining and activity assessment of activated sludge bacteria. Appl Environ Microbiol 68:4772–4779. https://doi.org/10.1128/AEM.68.10.4772-4779.2002
Goossens H, Ferech M, Vanderstichele R, Elseviers M (2005) Outpatient antibiotic use in Europe and association with resistance: a cross-national database study. Lancet 365:579–587. https://doi.org/10.1016/S0140-6736(05)70799-6
Gupta SK, Shin H, Han D, Hur HG, Unno T (2018) Metagenomic analysis reveals the prevalence and persistence of antibiotic- and heavy metal-resistance genes in wastewater treatment plant. J Microbiol 56:408–415. https://doi.org/10.1007/s12275-018-8195-z
Hakemi Vala M, Eyvazi S, Goudarzi H, Sarie HR, Gholami M (2016) Evaluation of clarithromycin resistance among iranian Helicobacter pylori isolates by E-test and real-time polymerase chain reaction methods. Jundishapur J Microbiol. https://doi.org/10.5812/jjm.29839
Halling-Sørensen B (2001) Inhibition of aerobic growth and nitrification of bacteria in sewage sludge by antibacterial agents. Arch Environ Contam Toxicol 40:451–460. https://doi.org/10.1007/s002440010197
Hardy DJ, Guay DRP, Jones RN (1992) Clarithromycin, a unique macrolide. Diagn Microbiol Infect Dis 15:39–53. https://doi.org/10.1016/0732-8893(92)90055-X
Hu Y, Yang X, Qin J, Lu N, Cheng G, Wu N, Pan Y, Li J, Zhu L, Wang X, Meng Z, Zhao F, Liu D, Ma J, Qin N, Xiang C, Xiao Y, Li L, Yang H, Wang J, Yang R, Gao GF, Wang J, Zhu B (2013) Metagenome-wide analysis of antibiotic resistance genes in a large cohort of human gut microbiota. Nat Commun 4:3151. https://doi.org/10.1038/ncomms3151
Hughes D, Andersson DI (2017) Environmental and genetic modulation of the phenotypic expression of antibiotic resistance. FEMS Microbiol Rev 41:374–391. https://doi.org/10.1093/femsre/fux004
Iamsawat S, Surawut S, Prammananan T, Leelaporn A, Jearanaisilavong J (2010) Multiplex PCR for detection of clarithromycin resistance and simultaneous species identification of Mycobacterium avium. Southeast Asian J Trop Med Public Heal 41:590–601
Jäger T, Hembach N, Elpers C, Wieland A, Alexander J, Hiller C, Krauter G, Schwartz T (2018) Reduction of antibiotic resistant bacteria during conventional and advanced wastewater treatment, and the disseminated loads released to the environment. Front Microbiol 9:1–16. https://doi.org/10.3389/fmicb.2018.02599
Kang J, Kang D (2016) International Journal of Food Microbiology Enhanced antimicrobial effect of organic acid washing against foodborne pathogens on broccoli by vacuum impregnation. Int J Food Microbiol 217:85–93. https://doi.org/10.1016/j.ijfoodmicro.2015.10.004
Kargar M, Doosti A, Ghorbani-Dalini S (2013) Detection of four clarithromycin resistance point mutations in Helicobacter pylori: comparison of real-time PCR and PCR-RFLP methods. Comp Clin Path 22:1007–1013. https://doi.org/10.1007/s00580-012-1519-1
Kitahara K, Yasutake Y, Miyazaki K (2012) Mutational robustness of 16S ribosomal RNA, shown by experimental horizontal gene transfer in Escherichia coli. Proc Natl Acad Sci 109:19220–19225. https://doi.org/10.1073/pnas.1213609109
Lebel M (1993) Pharmacokinetic properties of clarithromycin: a comparison with erythromycin and azithromycin. Can J Infect Dis 4:148–152
Leon G, Roy PH (2003) Excision and integration of cassettes by an integron integrase of Nitrosomonas europaea. J Bacteriol 185:2036–2041. https://doi.org/10.1128/JB.185.6.2036-2041.2003
Lépesová K, Kraková L, Pangallo D, Medveďová A, Olejníková P, Mackuľak T, Tichý J, Grabic R, Birošová L (2018) Prevalence of antibiotic-resistant coliform bacteria, Enterococcus spp. and Staphylococcus spp. in wastewater sewerage biofilm. J Glob Antimicrob Resist 14:145–151. https://doi.org/10.1016/j.jgar.2018.03.008
Li B, Qiu Y, Zhang J, Liang P, Huang X (2019a) Conjugative potential of antibiotic resistance plasmids to activated sludge bacteria from wastewater treatment plants. Int Biodeterior Biodegrad 138:33–40. https://doi.org/10.1016/j.ibiod.2018.12.013
Li R, Jay JA, Stenstrom MK (2019b) Fate of antibiotic resistance genes and antibiotic-resistant bacteria in water resource recovery facilities. Water Environ Res 91:5–20. https://doi.org/10.1002/wer.1008
Liu H-Q, Lam JCW, Li W-W, Yu H-Q, Lam PKS (2017) Spatial distribution and removal performance of pharmaceuticals in municipal wastewater treatment plants in China. Sci Total Environ 586:1162–1169. https://doi.org/10.1016/j.scitotenv.2017.02.107
López-Vázquez CM, Hooijmans CM, Brdjanovic D, Gijzen HJ, van Loosdrecht MCM (2008) Factors affecting the microbial populations at full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants in The Netherlands. Water Res 42:2349–2360. https://doi.org/10.1016/j.watres.2008.01.001
Loy A, Horn M, Wagner M (2003) Probebase: an online resource for rRNA-targeted oligonucleotide probes. Nucleic Acids Res 31:514–516. https://doi.org/10.1093/nar/gkv1232
Łuczkiewicz A, Felis E, Ziembinska A, Gnida A, Kotlarska E, Olanczuk-Neyman K, Surmacz-Gorska J (2013) Resistance of Escherichia coli and Enterococcus spp. to selected antimicrobial agents present in municipal wastewater. J Water Health 11:600–612. https://doi.org/10.2166/wh.2013.130
Łuczkiewicz A, Fudala-Książek S, Szopinska M, Jankowska K, Svahn O, Björklund E (2019) Fate of pharmaceuticals—via wastewater to the Baltic Sea. In: Proceedings of IWA 11th micropol and ecohazard conference, Seoul, Korea
McArdell CS, Molnar E, Suter MJF, Giger W (2003) Occurrence and fate of macrolide antibiotics in wastewater treatment plants and in the Glatt Valley watershed, Switzerland. Environ Sci Technol 37:5479–5486. https://doi.org/10.1021/es034368i
Novo A, André S, Viana P, Nunes OC, Manaia CM (2013) Antibiotic resistance, antimicrobial residues and bacterial community composition in urban wastewater. Water Res 47:1875–1887. https://doi.org/10.1016/j.watres.2013.01.010
Polikanov YS, Aleksashin NA, Beckert B, Wilson DN (2018) The mechanisms of action of ribosome-targeting peptide antibiotics. Front Mol Biosci 5:48. https://doi.org/10.3389/fmolb.2018.00048
Pray L (2008) Antibiotic resistance, mutation rates and MRSA. Natur Educ 1:30
Prestinaci F, Pezzotti P, Pantosti A (2015) Antimicrobial resistance: a global multifaceted phenomenon. Pathog Glob Health 109:309–318. https://doi.org/10.1179/2047773215Y.0000000030
Rizzo L, Manaia C, Merlin C, Schwartz T, Dagot C, Ploy MCC, Michael I, Fatta-Kassinos D (2013) Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci Total Environ 447:345–360. https://doi.org/10.1016/j.scitotenv.2013.01.032
Schmieder R, Edwards R (2012) Insights into antibiotic resistance through metagenomic approaches. Future Microbiol 7:73–89. https://doi.org/10.2217/fmb.11.135
Seyama S, Wajima T, Nakaminami H, Noguchi N (2016) Clarithromycin resistance mechanisms of epidemic β-lactamase-nonproducing ampicillin-resistant Haemophilus influenzae strains in Japan. Antimicrob Agents Chemother 60:3207–3210. https://doi.org/10.1128/AAC.00163-16
Shoemaker NB, Vlamakis H, Hayes K, Salyers AA (2001) Evidence for extensive resistance gene transfer among Bacteroides spp. and among bacteroides and other genera in the human colon. Appl Environ Microbiol 67:561–568. https://doi.org/10.1128/AEM.67.2.561-568.2001
Terada A, Usui H, Bao Q, Nakai S, Hosomi M (2017) Feasibility of biodegradation of clarithromycin by Nitrosomonas europaea. Kagaku Kogaku Ronbunshu 43:264–270. https://doi.org/10.1252/kakoronbunshu.43.264
Terzic S, Udikovic-Kolic N, Jurina T, Krizman-Matasic I, Senta I, Mihaljevic I, Loncar J, Smital T, Ahel M (2018) Biotransformation of macrolide antibiotics using enriched activated sludge culture: kinetics, transformation routes and ecotoxicological evaluation. J Hazard Mater 349:143–152. https://doi.org/10.1016/j.jhazmat.2018.01.055
Trebesius K, Panthel K, Strobel S, Vogt K, Faller G, Kirchner T, Kist M, Heesemann J, Haas R (2000) Rapid and specific detection of Helicobacter pylori macrolide resistance in gastric tissue by fluorescent in situ hybridisation. Gut 46:608–614. https://doi.org/10.1136/gut.46.5.608
UNESCO and HELCOM (2017) Pharmaceuticals in the aquatic environment of the Baltic Sea region—a status report. France, Paris
Von Wintersdorff CJH, Penders J, Van Niekerk JM (2016) Dissemination of antimicrobial resistance in microbial ecosystems through horizontal gene transfer. Front Microbiol 7:1–10. https://doi.org/10.3389/fmicb.2016.00173
Wagner M, Loy A (2002) Bacterial community composition and function in sewage treatment systems. Curr Opin Biotechnol 13:218–227. https://doi.org/10.1016/S0958-1669(02)00315-4
Wagner M, Hornt M, Daims H (2003) Fluorescence in situ hybridisation for the identification and characterisation of prokaryotes. Curr Opin Microbiol 6:302–309. https://doi.org/10.1016/S1369-5274(03)00054-7
Wang Z, Zhang XX, Huang K, Miao Y, Shi P, Liu B, Long C, Li A (2013) Metagenomic profiling of antibiotic resistance genes and mobile genetic elements in a tannery wastewater treatment plant. PLoS One 8:1–9. https://doi.org/10.1371/journal.pone.0076079
West BM, Liggit P, Clemans DL, Francoeur SN (2011) Antibiotic resistance, gene transfer, and water quality patterns observed in waterways near CAFO farms and wastewater treatment facilities. Water Air Soil Pollut 217:473–489. https://doi.org/10.1007/s11270-010-0602-y
WHO (2018) Antibiotic resistance. https://www.who.int/news-room/fact-sheets/detail/antibiotic-resistance. Accessed 21 Nov 2019
Yamagata A, Kato J, Hirota R, Kuroda A, Ikeda T, Takiguchi N, Ohtake H (1999) Isolation and characterization of two cryptic plasmids in the ammonia-oxidizing bacterium Nitrosomonas sp. strain ENI-11. J Bacteriol 181:3375–3381
Yang Y, Li B, Ju F, Zhang T (2013) Exploring variation of antibiotic resistance genes in activated sludge over a four-year period through a metagenomic approach. Environ Sci Technol 47:10197–10205. https://doi.org/10.1021/es4017365
Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214. https://doi.org/10.1089/10665270050081478
Zhou Z-C, Feng W-Q, Han Y, Zheng J, Chen T, Wei Y-Y, Gillings M, Zhu Y-G, Chen H (2018) Prevalence and transmission of antibiotic resistance and microbiota between humans and water environments. Environ Int 121:1155–1161
Ziembińska-Buczyńska A, Felis E, Folkert J, Meresta A, Stawicka D, Gnida A, Surmacz-Górska J (2015) Detection of antibiotic resistance genes in wastewater treatment plant – molecular and classical approach. Arch Environ Prot 41:23–32. https://doi.org/10.1515/aep-2015-0035
Zuckerman JM (2000) The newer macrolides: azithromycin and clarithromycin. Infect Dis Clin North Am 14:449–462. https://doi.org/10.1016/S0891-5520(05)70257-9
Acknowledgements
The research was financed by the Polish Ministry of Science and Higher Education under project entitled “Transfer of antibiotics resistance in activated sludge bacteria” (project no N N 523493134). Preparation of the article was financed by the Polish Ministry of Science and Higher Education (08/080/BK_18/0054). We want to thank Katarzyna Kunda for her valuable assistance during the laboratory part and for the FISH images.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
Rights and permissions
About this article
Cite this article
Gnida, A., Felis, E., Ziembińska-Buczyńska, A. et al. Evidence of mutations conferring resistance to clarithromycin in wastewater and activated sludge. 3 Biotech 10, 7 (2020). https://doi.org/10.1007/s13205-019-1989-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-019-1989-9