ISOLATION AND IDENTIFICATION OF FIMBRIAE AND TOXIN PRODUCTION BY ESCHERICHIA-COLI STRAINS FROM COWS WITH CLINICAL MASTITIS

A total of 20 Escherichia coli strains isolated from cases of bovine mastitis were examined for fimbriae production, for the presence of genes coding for enterotoxins (LT and ST1), verotoxins (VT), and for the production of cytotoxic necrotizing factors (CNF1 and CNF2). Fimbriae could be isolated from four strains. The N-terminal amino acid sequence of the fimbriae from two strains, was determined. The two sequences were almost identical and homologous to that of the major subunit of E. coli F17 fimbriae. A DNA probe was derived from this N-terminal sequence and used as probe in hybridization experiments with chromosomal DNA of the 20 strains. To lest if the strains contained genes that code for the F17 adhesin and the F17 major subunit, a Polymerase Chain Reaction (PCR) assay was performed with primers based on the nucleotide sequence of the genes. Eleven of 20 strains contained sequences that were homologous with sequences for the F17 fimbrial subunit and the F17 adhesin. Strains were tested directly for toxin production on Hela cells and by PCR for the presence of toxin genes. One of the twenty strains, produced a CNF toxin. No strains reacted positive in the PCR for LT, ST1 and verotoxin genes.