Faecal indicator bacteria and antibiotic-resistant β-lactamase producing Escherichia coli in blackwater: a pilot study

  • Urška Šunta Department of Sanitary Engineering, Faculty of Health Sciences, University of Ljubljana, Ljubljana
  • Miha Žitnik Department of Sanitary Engineering, Faculty of Health Sciences, University of Ljubljana, Ljubljana
  • Noemi Concetta Finocchiaro Department of Agriculture, Food and Environment, University of Catania, Catania
  • Tjaša Griessler Bulc Department of Sanitary Engineering, Faculty of Health Sciences, University of Ljubljana, Ljubljana
  • Karmen Godič Torkar University in Ljubljana, Faculty of Health Sciences, Ljubljana
Keywords: antimicrobial resistance, extended spectrum β-lactamases, metallo-β-lactamases, public health, wastewater treatment

Abstract

The aim of this study was to identify and quantify faecal indicator bacteria in blackwater collected from a source separation unit and determine the amount of E. coli isolates resistant to antimicrobials and their potential to produce extended spectrum β-lactamases (ESβLs) and metallo-β-lactamases (MβLs), which hydrolyse the most important antibiotics used in clinical practice. Most of the isolates were resistant to amoxicillin with clavulanic acid (36.4 %), followed by ticarcillin with clavulanic acid (22.7 %) and tetracycline (18.2 %). ESβL-producing genes blaCTX-M and blaTEM were found in three (13.6 %) and four (18.2 %) E. coli strains, respectively, while MβL genes were found in two (9.1 %). By separating at source, this pilot study clearly shows that gastrointestinal bacteria of healthy people can be an important source of antibiotic resistance released into the environment through wastewaters. One way to prevent that is to treat wastewater with a combination of TiO2, UV light, or ozone, as successful methods to remove resistant bacteria and prevent their spread in the environment.

Author Biography

Karmen Godič Torkar, University in Ljubljana, Faculty of Health Sciences, Ljubljana

Department for Sanitary Engeneering

References

Meinzinger F and Oldenburg M. Characteristics of source-separated household wastewater flows: a statistical assessment. Water Sci Technol 2009;59:1785–91. doi: 10.2166/wst.2009.185

Oarga-Mulec A, Jenssen PD, Krivograd Klemenčič A, Uršič M, Griessler Bulc T. Zero-discharge solution for blackwater treatment at remote tourist facilities. J Clean Prod 2017;166:798–805. doi: 10.1016/j.jclepro.2017.08.002

World Health Organization (WHO). Guidelines for the safe use of wastewater, excreta and greywater. Volume 4: Excreta and greywater use in agriculture. 4th ed. Geneva: WHO; 2006.

Oarga A, Griessler Bulc T, Jenssen DP, Mulec J. Monitoring of microbial indicator groups in organically heavily loaded wastewater treatment systems by using Rida®Count kits. Fresen Environ Bull 2012;21:3886–93.

Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA. Diversity of the human intestinal microbial flora. Science 2005;308:1635–8. doi: 10.1126/science.1110591

Bessa LJ, Barbosa-Vasconcelos A, Mendes A, Vaz-Pires P, Martins da Costa P. High prevalence of multidrug-resistant Escherichia coli and Enterococcus spp. in river water, upstream and downstream of a wastewater treatment plant. J Water Health 2014;12:426–35. doi: 10.2166/wh.2014.160

Office of Legislation Republic of Slovenia. Uredba o odvajanju in čiščenju komunalne odpadne vode Decree on the discharge and treatment of urban wastewater, in Slovenian. Uradni list Republike Slovenije 2015;98:12234.

Wendland C. Anaerobic digestion of blackwater and kitchen refuse dissertation. Hamburg: Institute of Wastewater Management and Water Protection, Technical University of Hamburg-Harburg; 2008.

Korzeniewska E, Korzeniewska A, Harnisz M. Antibiotic resistant Escherichia coli in hospital and municipal sewage and their emission to the environment. Ecotox Environ Saf 2013;91:96–102. doi: 10.1016/j.ecoenv.2013.01.014

Huddleston JR. Horizontal gene transfer in the human gastrointestinal tract: potential spread of antibiotic resistance genes. Infect Drug Resist 2014;7:167–76. doi: 10.2147/IDR.S48820

Bush K, Jacoby GA. Updated functional classification of β-lactamases. Antimicrob Agents Chemoth 2010;54:969–76. doi: 10.1128/AAC.01009-09

Woodford N, Ward ME, Kaufmann ME, Turton J, Fagan EJ, James D, Johnson AP, Pike R, Warner M, Cheasty T, Pearson A, Harry S, Leach JB, Loughrey A, Lowes JA, Warren RE, Livermore DM. Community and hospital spread of Escherichia coli producing CTX-M extended-spectrum beta-lactamases in the UK. J Antimicrob Chemother 2004;54:735–43. doi: 10.1093/jac/dkh424

Reinthaler FF, Feierl G, Galler H, Haas D, Leitner E, Mascher F, Melkes A, Posch J, Winter I, Zarfel G, Marth E. ESBL-producing E. coli in Austrian sewage sludge. Water Res 2010;44:1981–5. doi: 10.1016/j.watres.2009.11.052

ISO 19458:2006. Water Quality - Sampling for microbiological analysis. Brussels: International Organization for Standardization; 2006.

ISO 8199:2005. Water Quality - General guidance on the enumeration of microorganisms by culture. Brussels: International Organization for Standardization; 2005.

Finney M, Smullen J, Foster HA, Brokx S, Storey DM. Evaluation of Chromocult coliform agar for the detection and enumeration of Enterobacteriaceae from faecal samples from healthy subjects. J Microbiol Methods 2003;54:353–8. doi: 10.1016/S0167-7012(03)00068-X

ISO 9308–1:2014. Water Quality - Enumeration of Escherichia coli and coliform bacteria - Part 1: Membrane filtration method for waters with low bacterial background flora. Brussels: International Organization for Standardization; 2014.

ISO 7899–2:2000. Water Quality - Detection and enumeration of intestinal enterococci - Part 2: Membrane filtration method. Brussels: International Organization for Standardization; 2000.

Merck. Mannitol salt phenol-red agar [displayed 3 April 2019]. Available at http://www.merckmillipore.com/INTL/en/product/Mannitol-salt-phenol-red-agar,MDA_CHEM-105404#anchor_DS

ISO 6461–2:2013. Water quality - Detection and enumeration of the spores of sulphite-reducing anaerobes (clostridia) - Part 2: Method by membrane filtration. Brussels: International Organization for Standardization; 2013.

Ferens WA, Hovde CJ. Escherichia coli O157:H7: animal reservoir and sources of human infection. Foodborne Pathog Dis 2011;8:465–87. doi: 10.1089/fpd.2010.0673

Clinical and Laboratory Standards Institute (CLSI). M100-S23 Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Third Informational Supplement. Wayne (PA): CLSI; 2013.

Queipo-Ortuño MI, Colmenero J de D, Macias M, Bravo MJ, Morata P. Preparation of bacterial DNA template by boiling and effect of immunoglobulin G as an inhibitor in real-time PCR for serum samples from patients with brucellosis. Clin Vaccine Immunol 2008;15:293–6. doi: 10.1128/CVI.00270-07

Arlet G, Brami G, Decre D, Flippo A, Gaillot O, Lagrange PH, Philippon A. Molecular characterization by PCR-restriction fragment polymorphism of TEM beta-lactamases. FEMS Microbiol Lett 1995;134:203–8. doi: 10.1111/j.1574-6968.1995.tb07938.x

Saladin M, Cao VT, Lambert T, Donay JL, Herrmann JL, Ould-Hocine Z, Verdet C, Delisle F, Philippon A, Arlet G. Diversity of CTX-M β-lactamases and their promoter regions from Enterobacteriaceae isolated in three Parisian hospitals. FEMS Microbiol Lett 2002;209:161–8. doi: 10.1111/j.1574-6968.2002.tb11126.x

Ellington MJ, Kistler J, Livermore DM, Woodford N. Multiplex PCR for rapid detection of genes encoding acquired metallo-beta-lactamases. J Antimicrob Chemother 2007;59:321–2. doi: 10.1093/jac/dkl481

Woodford N. Rapid characterization of beta-lactamases by multiplex PCR. Methods Mol Biol 2010;642:181–92. doi: 10.1007/978-1-60327-279-7_14

Ktari S, Arlet G, Mnif B, Gautier V, Mahjoubi F, Ben Jmeaa M, Bouaziz M, Hammami A. Emergence of multidrug-resistant Klebsiella pneumoniae isolates producing VIM-4 metallo-beta-lactamase, CTX-M-15 extended-spectrum beta-lactamase, and CMY-4 AmpC beta-lactamase in a Tunisian university hospital. Antimicrob Agents Chemother 2006;50:4198–201. doi: 10.1128/AAC.00663-06

Acton DS, Tempelmans Plat-Sinnige MJ, van Wamel W, de Groot N, van Belkum A. Intestinal carriage of Staphylococcus aureus: how does its frequency compare with that of nasal carriage and what is its clinical impact? Eur J Clin Microbiol Infect Dis 2009;28:115–27. https://doi.org/10.1007/s10096-008-0602-7

Akinkunmi EO, Adeyemi OI, Igbeneghu OA, Olaniyan EO, Omonisi AE, Lamikanra A. The pathogenicity of Staphylococcus epidermidis on the intestinal organs of rats and mice: an experimental investigation. BMC Gastroenterol 2014;14:126–34. doi: 10.1186/1471-230X-14-126

Islam MMM, Hofstra N, Islam MA. The Impact of environmental variables on faecal indicator bacteria in the Betna River Basin, Bangladesh. Environ Processes 2017;4(2):319–32. https://doi.org/10.1007/s40710-017-0239-6

Hardalo C, Edberg SC. Pseudomonas aeruginosa: Assessment of risk from drinking water. Crit Rev Microbiol 1997;23:47–75. doi: 10.3109/10408419709115130

Laughlin RS, Musch MW, Hollbrook CJ, Rocha FM, Chang EB, Alverdy JC. The key role of Pseudomonas aeruginosa PA-I lectin on experimental gut-derived sepsis. Ann Surg 2000;232:133–42. doi: 10.1097/00000658-200007000-00019

Engelkirk PG, Duben-Engelkirk J. Chapter 12. Gram-negative bacilli: The family Enterobacteriaceae. In: Laboratory diagnosis of infectious diseases: essentials of diagnostic microbiology. Philadelphia: Lippincott Williams & Wilkins; 2008. p. 292–318.

Janda JM, Abbott SL. The Genus Hafnia: from soup to nuts. Clin Microbiol Rev 2006;19:12–8. doi: 10.1128/CMR.19.1.12-28.2006

Vinué L, Sáenz Y, Martínez S, Somalo S, Moreno MA, Torres C, Zarazaga M. Prevalence and diversity of extended-spectrum β-lactamases in faecal Escherichia coli isolates from healthy humans in Spain. Clin Microbiol Infect 2009;15:954–7. doi: 10.1111/1469-0691.2009.02803.x

Rodríguez-Baño J, López-Cerero L, Navarro MD, de Alba PD, Pascual A. Faecal carriage of extended-spectrum β-lactamase-producing Escherichia coli: prevalence, risk factors and molecular epidemiology. J Antimicrob Chemother 2008;62:1142–9. doi: 10.1093/jac/dkn293

Korzeniewska E, Harnisz M. Extended-spectrum beta-lactamase (ESβL)-positive Enterobacteriaceae in municipal sewage and their emission to the environment. J Environ Manage 2013;128:904–11. doi: 10.1016/j.jenvman.2013.06.051

Osínska A, Korzeniewska E, Harnisz M, Niestępski S. The prevalence and characterization of antibiotic-resistant and virulent Escherichia coli strains in the municipal wastewater system and their environmental fate. Sci Total Environ 2017;577:367–75. doi: 10.1016/j.scitotenv.2016.10.203

Woodford N, Carattoli A, Karisik E, Underwood A, Ellington JM, Livermore DM. Complete nucleotide sequences of plasmids pEK204, pEK499, and pEK516, encoding CTX-M enzymes in three major Escherichia coli lineages from the United Kingdom, all belonging to the international O25:H4-ST131 clone. Antimicrob Agents Chemother 2009;53:4472–82. doi: 10.1128/AAC.00688-09

Öncü NB, Menceloğlu YZ, Balcıoğlu IA. Comparison of the effectiveness of chlorine, ozone, and photocatalytic disinfection in reducing the risk of antibiotic resistance pollution. J Adv Oxid Technol 2016;14:196–203. doi: 10.1515/jaots-2011-0203

Karaolia P, Michel-Kordatou I, Hapeshi E, Drosou C, Bertakis Y, Christofilos D, Armatas GS, Sygellou L, Schwartz T, Xekoukoulotakis NP, Fatta-Kassinos D. Removal of antibiotics, antibiotic resistant bacteria and their associated genes by graphene-based TiO2 composite photocatalysts under solar radiation in urban wastewaters. Appl Catal B 2018;224:810–24. doi: 10.1016/j.apcatb.2017.11.020c

Ren S, Boo C, Guo N, Wang S, Elimelech M, Wang Y. Photocatalytic reactive ultrafiltration membrane for removal of antibiotic resistant bacteria and antibiotic resistance genes from wastewater effluent. Environ Sci Technol 2018:52:8666–73. doi: 10.1021/acs.est.8b01888

Published
2019-06-07
How to Cite
1.
Šunta U, Žitnik M, Finocchiaro NC, Griessler Bulc T, Godič Torkar K. Faecal indicator bacteria and antibiotic-resistant β-lactamase producing Escherichia coli in blackwater: a pilot study. Arh Hig Rada Toksikol [Internet]. 2019Jun.7 [cited 2024Apr.23];70(2). Available from: https://arhiv.imi.hr/index.php/arhiv/article/view/1047
Section
Original article