A survey of extended-spectrum β-lactamase-producing Enterobacteriaceae in environmental water in Okinawa Prefecture of Japan and relationship with indicator organisms
Surveys of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-pE) in stream water and untreated wastewater were carried out in Okinawa Prefecture, Japan. Thirty-six samples of water were collected from 18 streams in Okinawa Prefecture, as well as ten samples of wastewater flowing into four wastewater treatment plants (WWTPs). We investigated bacterial species, Escherichia coli O antigen, ESBL phenotype, ESBL genotype, and pulsed-field gel electrophoresis (PFGE) type of isolates, and total viable count and fecal coliforms as indicator organisms. The relation between indicator organisms and ESBL-pE was also validated using the same samples. A total of 141 ESBL-pE including 107 E. coli, 15 Klebsiella pneumoniae, 2 Proteus mirabilis, and 17 other species was isolated from stream water and wastewater. Of the 141 ESBL-pE, 14.9% and 54.6% were found to be blaCTX-M-15 and blaCTX-M-14-like types, respectively, which have been found in hospital isolates in Okinawa. Two pairs of possibly related patterns according to PFGE criteria were isolated from stream water and wastewater in two districts. When ESBL-pE was significantly isolated, total viable count and fecal coliform boundaries were ≥ 6.0 × 103 CFU/ml and ≥ 4.3 × 102 most probable number/100 ml, respectively. These results suggested that ESBL-pE isolated from stream water is human derived, and that total viable count and fecal coliforms will be useful as indicators for confirming the spread of ESBL-pE to the environment by means of simple hygiene surveys.
KeywordsESBL-producing Enterobacteriaceae Survey Stream water Wastewater Indicator organism Relationship
We thank the staff of Okinawa Prefecture Sewerage Management Office who allowed us to conduct the research and provided access to wastewater.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Boyd DA, Tyler S, Christianson S, McGeer A, Muller MP, Willey BM, Bryce E, Gardam M, Nordmann P, Mulvey MR, the Canadian Nosocomial Infection Surveillance Program, Health Canada (2004) Complete nucleotide sequence of a 92-kilobase plasmid harboring the CTX-M-15 extended-spectrum beta-lactamase involved in an outbreak in long-term-care facilities in Toronto, Canada. Antimicro Agents Chemother 48:3758–3764CrossRefGoogle Scholar
- Chia J-H, Chu C, Su LH, Chiu C-H, Kuo A-J, Sun C-F, Wu T-L (2005) Development of a multiplex PCR and SHV melting-curve mutation detection system for detection of some SHV and CTX-M β-lactamases of Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae in Taiwan. J Clin Microbiol 43:4486–4491CrossRefGoogle Scholar
- Clermont O, Dhanji H, Upton M, Gibreel T, Fox A, Boyd D, Mulvey MR, Nordmann P, Ruppé E, Sarthou JL, Frank T, Vimont S, Arlet G, Branger C, Woodford N, Erick Denamur E (2009) Rapid detection of the O25b-ST131 clone of Escherichia coli encompassing the CTX-M-15-producing strains. J Antimicrob Chemother 64:274–277CrossRefGoogle Scholar
- Conte D, Palmeiro JK, da Silva Nogueira K, de Lima TMR, Cardoso MA, Pontarolo R, Degaut Pontes FL, Dalla-Costa LM (2017) Characterization of CTX-M enzymes, quinolone resistance determinants, and antimicrobial residues from hospital sewage, wastewater treatment plant, and river water. Ecotoxicol Environ Saf 136:62–69CrossRefGoogle Scholar
- Farmer III JJ, Boatwright KD, Michael J (2007) 42 Enterobacteriaceae: introduction and identification. In: Murray PR (ed) Manual of clinical microbiology, Vol. 1, 9th edn. ASM, Washington D.C., pp 649–669Google Scholar
- Japan Ministry of the Environment (1993) Article 16, standard of environment, environmental standards lawGoogle Scholar
- Kittinger C, Lipp M, Folli B, Kirschner A, Baumert R, Galler H, Grisold AJ, Luxner J, Weissenbacher M, Farnleitner AH, Zarfel G (2016) Enterobacteriaceae isolated from the River Danube: antibiotic resistances, with a focus on the presence of ESBL and carbapenemases. PLoS One 11:e0165820CrossRefGoogle Scholar
- Nakama R, Shingaki A, Miyazato H, Higa R, Nagamoto C, Hamamoto K, Ueda S, Hachiman T, Touma Y, Miyagi K, Kawahara R, Toyosato T, Hirai I (2016) Current status of extended spectrum β-lactamase-producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis in Okinawa Prefecture, Japan. J Infect Chemother 22:281–286CrossRefGoogle Scholar
- Nakamura T, Komatsu M, Yamasaki K, Fukuda S, Miyamoto T, Higuchi T, Ono T, Nishio H, Sueyoshi N, Kida K, Satoh K, Toda H, Toyokawa M, Nishi I, Sakumoto M, Akagi M, Nakai I, Kofuku T, Orita T, Wada Y, Zikimoto T, Koike C, Kinoshita S, Hirai I, Takahashi H, Matsumura N, Yamamoto Y (2012) Epidemiology of Escherichia coli, Klebsiella species, and Proteus mirabilis strains producing extended-spectrum β-lactamase from clinical samples in the Kinki region of Japan. Am J Clin Pathol 137:620–626CrossRefGoogle Scholar
- Nakamura A, Komatsu M, Noguchi N, Ohno Y, Hashimoto E, Matsutani H, Abe N, Fukuda S, Kohno H, Nakamura F, Matsuo S, Kawano S (2016) Analysis of molecular epidemiologic characteristics of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli colonizing feces in hospital patients and community dwellers in a Japanese city. J Infect Chemother 22:102–107CrossRefGoogle Scholar
- Paterson DL, Hujer KM, Hujer AM, Yeiser B, Bonomo MD, Rice LB, Bonomo RA, the International Klebsiella Study Group (2003) Extended-spectrum β-lactamases in Klebsiella pneumoniae bloodstream isolates from seven countries: dominance and widespread prevalence of SHV- and CTX-M-type β-lactamases. Antimicro Agents Chemother 47:3554–3560CrossRefGoogle Scholar
- Randall LP, Lodge MP, Elviss NC, Lemma FL, Hopkins KL, Teale CJ, Woodford N (2017) Evaluation of meat, fruit and vegetables from retail stores in five United Kingdom regions as sources of extended-spectrum beta-lactamase (ESBL)-producing and carbapenem-resistant Escherichia coli. Int J Food Microbiol 241:283–290CrossRefGoogle Scholar
- Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B (1995) Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33:2233–2239Google Scholar
- The European Parliament and The Council (2006) Directive 2006/7/EC. In: The European UnionGoogle Scholar
- United States Environmental Protection Agency (2012) Office of Water EPA-820-F-12-061, 2012 recreational water quality criteriaGoogle Scholar
- Ye Q, Wu Q, Zhang S, Zhang J, Yang G, Wang H, Huang J, Chen M, Xue L, Wang J (2017) Antibiotic-resistant extended spectrum ß-lactamase- and plasmid-mediated AmpC-producing Enterobacteriaceae isolated from retail food products and the Pearl River in Guangzhou, China. Front Microbiol 8:96Google Scholar
- Zarfel G, Lipp M, Gürtl E, Folli B, Baumert R, Kittinger C (2017) Troubled water under the bridge: screening of River Mur water reveals dominance of CTX-M harboring Escherichia coli and for the first time an environmental VIM-1 producer in Austria. Sci Total Environ 593-594:399–405CrossRefGoogle Scholar