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In this work, by high-throughput sequencing, antibiotic resistance genes, including class A ( bla   CTX-M ,  bla   Z ,  bla   TEM ,  bla   VEB ,  bla   KLUC , and  bla   SFO ), class C ( bla   SHV ,  bla   DHA ,  bla   MIR ,  bla   AZECL-29 , and  bla   ACT ), and class D ( bla   OXA )  β -lactamase genes, were identified among the pooled genomic DNA from 212 clinical  Enterobacter cloacae  isolates. Six  bla   MIR -positive  E. cloacae  strains were identified, and pulsed-field gel electrophoresis (PFGE) showed that these strains were not clonally related. The complete genome of the  bla   MIR   - positive strain (Y546) consisted of both a chromosome (4.78 Mb) and a large plasmid pY546 (208.74 kb). The extended-spectrum  β -lactamases (ESBLs) ( bla   SHV-12  and  bla   CTX-M-9a ) and AmpC ( bla   MIR ) were encoded on the chromosome, and the pY546 plasmid contained several clusters of genes conferring resistance to metals, such as copper ( pco ), arsenic ( ars ), tellurite ( ter ), and tetrathionate ( ttr ), and genes encoding many divalent cation transporter proteins. The comparative genomic analyses of the whole plasmid sequence and of the heavy metal resistance gene-encoding regions revealed that the plasmid sequences of  Klebsiella pneumoniae  (such as pKPN-332, pKPN-3967, and pKPN-262) shared the highest similarity with those of pY546. It may be concluded that a variety of  β -lactamase genes present in  E. cloacae  which confer resistance to  β -lactam antibiotics and the emergence of plasmids carrying heavy metal resistance genes in clinical isolates are alarming and need further surveillance.

The β-Lactamase Gene Profile and a Plasmid-Carrying Multiple Heavy Metal Resistance Genes of Enterobacter cloacae