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Ecotoxicology

, Volume 22, Issue 2, pp 402–414 | Cite as

Molecular diversity of class 2 integrons in antibiotic-resistant gram-negative bacteria found in wastewater environments in China

  • Ruirui Xia
  • Ye Ren
  • Xianhu Guo
  • Hai XuEmail author
Article

Abstract

The molecular architecture of class 2 integrons among gram-negative bacteria from wastewater environments was investigated in Jinan, China. Out of the 391 antibiotic-resistant bacteria found, 38 isolates harboring class 2 integrons encoding potentially transferrable genes that could confer antibiotic resistance were found. These isolates were classified into 19 REP-PCR types. These strains were identified using 16S rRNA gene sequencing and found to be as follows: Proteus mirabilis (16), Escherichia coli (7), Providencia spp. (7), Proteus spp. (2), P. vulgaris (3), Shigella sp. (1), Citrobacter freundii (1), and Acinetobacter sp. (1). Their class 2 integron cassette arrays were amplified and then either analyzed using PCR–RFLP or sequenced. The typical array dfrA1-sat2-aadA1 was detected in 27 isolates. Six atypical arrays were observed, including three kinds of novel arrangements (linF2(∆attC1)-dfrA1(∆attC2)-aadA1-orf441 or linF2(∆attC1)-dfrA1(∆attC2)-aadA1, dfrA1-catB2-sat2-aadA1, and estX Vr -sat2-aadA1) and a hybrid with the 3′CS of class 1 integrons (dfrA1-sat2-aadA1-qacH), and dfrA1-sat1. Twenty-four isolates were also found to carry class 1 integrons with 10 types of gene cassette arrays. Several non-integron-associated antibiotic resistance genes were found, and their transferability was investigated. Results showed that water sources in the Jinan region harbored a diverse community of both typical and atypical class 2 integrons, raising concerns about the overuse of antibiotics and their careless disposal into the environment.

Keywords

Antibiotic-resistant Hybrid integron Multigene cassette Non-integron associated antibiotic resistance gene Novel gene cassette arrays 

Notes

Acknowledgments

This work was supported by the grants from the National Natural Science Foundation of China (30870084 and 31170112), the Medical and Health Technology Foundation of Shandong (2011HZ050), and the State Key Laboratory of Microbial Technology, Shandong University.We would like to thank Professor Julian Davies and Dr. Vivian Miao from the University of British Columbia for helpful comments on manuscript revision and George A. Jacoby for providing E. coli J53 (Azide R).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10646_2012_1034_MOESM1_ESM.doc (5.1 mb)
Supplementary material 1 (DOC 5237 kb)

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  1. 1.State Key Laboratory of Microbial TechnologySchool of Life Science, Shandong UniversityJinanChina
  2. 2.Shandong Longlive Bio-technology Co. Ltd., High-tech ZoneYucheng, DezhouChina

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