Ribotyping and plasmid profiling of Vibrio anguillarum serovar O2 and Vibrio ordalii

Abstract

One hundred and twenty-nine strains of Vibrio anguillarum serovar O2 and 14 strains of Vibrio ordalii were ribotyped and examined for plasmid contents. A total of 35 different ribotypes were detected. The V. anguillarum serovar O2 strains were divided into 32 different ribotypes. The V. ordalii strains showed three different ribotypes, clearly distinct from those of the V. anguillarum strains. Ribotypes were separated into seven clusters, of which one comprised the V. ordalii strains. Clustering of the strains indicated a genetic difference between North European and South European V. anguillarum O2 strains. Sero-subgroups O2a and O2b shared ribotypes; however, three of the clusters did not include O2a strains. All V. ordalii strains had a plasmid of 32 kb. This plasmid was not detected in any of the V. anguillarum strains. Seventeen different plasmid profiles with 17 different sized plasmids were detected among the V. anguillarum strains. Most of the plasmids were small (< 6 kb) and found in several strains. Except for one South European strain, plasmids were detected only in the North European strains of V. anguillarum O2.

Plasmid profiles, restriction fragment length polymorphisms and O-serotypes amongVibrio anguillarumisolates

“…This figure is somewhat lower than previously reported by Larsen and Olsen [13] who found that 54 out of 103 (52%) 02 strains carried plasmids. Tiainen and colleagues [33] detected plasmids in 48 out of 129 (37 %) 02 strains, a figure comparable to that reported in the present study. However, Tiainen and colleagues [33] found a remarkable difference between northern European and southern European strains.…”

“…Tiainen and colleagues [33] detected plasmids in 48 out of 129 (37 %) 02 strains, a figure comparable to that reported in the present study. However, Tiainen and colleagues [33] found a remarkable difference between northern European and southern European strains. Plasmids were almost exclusively found in the northern European isolates, and since in the study by Larsen and Olsen [13] only Danish isolates were included, this may very well account for the reported difference in ,,t tr _ ) Rt C' £, _" _q (7 plasmid content.…”

Restriction fragment length polymorphism of the pMJ101-like plasmid and ribotyping in the fish pathogenVibrio ordalii

“…A plasmid of approximately 32 kb has been detected in all V. ordalii strains studied so far [3,5,8]. The plasmid of the V. ordalii type strain, DF3K, has been designated pMJ101 [1].…”

“…In addition, the clinical symptoms and pathological changes caused by this organism in fish are different from those recorded for V. anguillarum [7]. It was recently demonstrated that V. ordalii and V. anguillarum were easily distinguished by ribotyping [8]. V. ordalii cross-reacts serologically with V. anguillarum serogroup 02a [9] although minor antigenic differences have been demonstrated.…”

“…Briefly, chromosomal DNA was extracted using an AutoGen 540 (AutoGen Instruments, Bioclear, Potterne, UK) automate, and 2-5 ,ug DNA were digested with Mlu I (Pharmacia, SaintQuentin-Yvelines, France). Fragments were separated by agarose gel electrophoresis in TBE buffer (tris 89 mm, boric acid 89 mM, EDTA 2 mM, pH 8 3) and blotted onto nylon membranes (Hybond-N, Amersham International, Amersham, England) using a VacuGene System (Pharmacia). After transfer, DNA was fixed to the membranes by baking at 80°C for 15 min and hybridized with an acetylaminofluorene (AAF)-labelled 16S and 23S rRNA probe from E. coli (Eurogentec, Seraing, Belgium) as described by Grimont and colleagues [26].…”