J Vet Sci.  2008 Mar;9(1):75-83. 10.4142/jvs.2008.9.1.75.

The expression of plasmid mediated afimbrial adhesin genes in an avian septicemic Escherichia coli strain

• Stehling E
• Campos TA
• Brocchi M
• de Carvalho Azevedo VA
• da Silveira WD

• KMID: 1102956
• DOI: http://doi.org/10.4142/jvs.2008.9.1.75

Abstract

An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD(50) of 4.0 x 10(5) CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin. The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa. Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD(50) of 1.2 x 10(12) CFU/ml. All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid. The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively. These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion. The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes.

Keyword

adhesion; avian; Escherichia coli; plasmids

MeSH Terms

Adhesins, Escherichia coli/*genetics/*metabolism
Animals
Chickens
Escherichia coli/genetics/*metabolism
Gene Expression Regulation, Bacterial
Plasmids/*genetics/metabolism
Poultry Diseases/*microbiology
Sepsis/*microbiology

Figure

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  Fig. 1 Agarose gel electrophoresis (0.7%) of plasmid DNA from the SEPT13 strain, its derivative transformant strains and the reference plasmids Lane 1: Strain V517 (32 MDa), Lane 2: Strain SEPT13, Lane 3: Strain ST16, Lane 4: Recipient strain MS101 harboring the 93 MDa plasmid (Strain transformant A).

• 

  Fig. 2 Adhesion of strain SEPT13 and its derivative transformant strains to Hep-2 cells. (A) Strain SEPT 13; (B) Strain MS101 (C) Strain ST16; (D) Strain transformant A; (E) Strain HB101; (f) Strain transformant B. ×1,000.

• 

  Fig. 3 Adhesion of strains SEPT 13 and its derivative transformant strains to tracheal epithelial cells. (A) Strain SEPT 13; (B) Recipient strain MS101; (C) Strain ST16; (D) Recipient strain MS101 harboring the 93 MDa plasmid (Strain transformant A). Arrowheads identify bacterial cells adherent to the tracheal epithelial cells. ×1,000.

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  Fig. 4 Electron microscopy studies of fimbria expression by the E. coli strains. (A) Strain SEPT 13, ×32,000; (B) Recipient strain HB101, ×80,000; (C) Recipient strain HB101 harboring the 93MDa plasmid (Strain transformant B), ×40,000; (D) Strain MS101, ×18,000.

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