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Applied and Environmental Microbiology, March 2007, p. 1976-1983, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02171-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Plasmid Replicon Typing of Commensal and Pathogenic Escherichia coli Isolates{triangledown}

Timothy J. Johnson,1 Yvonne M. Wannemuehler,1 Sara J. Johnson,1 Catherine M. Logue,2 David G. White,3 Curt Doetkott,2 and Lisa K. Nolan1*

Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, 1802 Elwood Drive, VMRI #2, Iowa State University, Ames, Iowa 50011,1 U.S. Food and Drug Administration, Center for Veterinary Medicine, Office of Research, 8401 Muirkirk Rd., Laurel, Maryland 20708,2 Information Technology Services, North Dakota State University, Fargo, North Dakota 581053

Received 15 September 2006/ Accepted 20 January 2007

Despite the critical role of plasmids in horizontal gene transfer, few studies have characterized plasmid relatedness among different bacterial populations. Recently, a multiplex PCR replicon typing protocol was developed for classification of plasmids occurring in members of the Enterobacteriaceae. Here, a simplified version of this replicon typing procedure which requires only three multiplex panels to identify 18 plasmid replicons is described. This method was used to screen 1,015 Escherichia coli isolates of avian, human, and poultry meat origin for plasmid replicon types. Additionally, the isolates were assessed for their content of several colicin-associated genes. Overall, a high degree of plasmid variability was observed, with 221 different profiles occurring among the 1,015 isolates examined. IncFIB plasmids were the most common type identified, regardless of the source type of E. coli. IncFIB plasmids occurred significantly more often in avian pathogenic E. coli (APEC) and retail poultry E. coli (RPEC) than in uropathogenic E. coli (UPEC) and avian and human fecal commensal E. coli isolates (AFEC and HFEC, respectively). APEC and RPEC were also significantly more likely than UPEC, HFEC, and AFEC to possess the colicin-associated genes cvaC, cbi, and/or cma in conjunction with one or more plasmid replicons. The results suggest that E. coli isolates contaminating retail poultry are notably similar to APEC with regard to plasmid profiles, with both generally containing multiple plasmid replicon types in conjunction with colicin-related genes. In contrast, UPEC and human and avian commensal E. coli isolates generally lack the plasmid replicons and colicin-related genes seen in APEC and RPEC, suggesting limited dissemination of such plasmids among these bacterial populations.

* Corresponding author. Mailing address: Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, 1802 Elwood Drive, VMRI #2, Iowa State University, Ames, IA 50011. Phone: (515) 294-3470. Fax: (515) 294-3839. E-mail: lknolan{at}iastate.edu.

{triangledown} Published ahead of print on 2 February 2007.

Applied and Environmental Microbiology, March 2007, p. 1976-1983, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02171-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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