4.5 Article

The identification and determination of toxin genes of Vibrio strains causing hemorrhagic disease on red drum (Sciaenops ocellatus) using PCR

Journal

AMB EXPRESS
Volume 11, Issue 1, Pages -

Publisher

SPRINGER
DOI: 10.1186/s13568-020-01161-w

Keywords

Sciaenops ocellatus; TDH; TRH; TLH; ToxR; Toxin gene; Vibrio

Funding

  1. Hue University
  2. Vietnam Ministry of Education and Training [CT-2018-DHH-07]

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Data was collected from 30 strains of Vibrio sampled from different organs of Sciaenops ocellatus infection. Results showed high similarity in 16S rRNA region nucleotide sequences with published species, and the presence of toxin genes in most Vibrio strains. Six parameters were used to measure DNA polymorphism in this population, revealing high diversity. A phylogenetic tree divided the strains into six groups based on their genetic differences. The study also identified the presence of specific Vibrio species on different organs of the infected fish.
Data were collected from 30 strains of Vibrio and sampled on different organs (brain, hemorrhagic site and digestive tract) of Sciaenops ocellatus infection. The results showed that the nucleotide sequences 16S rRNA region are highly similar to those of V. alginolyticus, V. azureus, V. fluvialis, V. natriengens and V. orientalis, which were published on Genbank and other, ranging from 98.05 to 100%. The digestive tract has the most common Vibrio strains (V. alginolyticus [16] V. azureus [7] and V. fluvialis). Thereout, 25 of 30 strains of Vibrio contained 1 to 3 toxin genes, except V. parahaemolyticus. Six parameters were used to measure the DNA polymorphism of 33 homologous DNA sequences in this Vibrio bacteria population. The results indicated that number of separate polymorphic sites (S), total number of mutant sites (Eta), number of haplotype (h), haplotype diversity (Hd), average number of nucleotide differences (k), nucleotide diversity (Pi) were 98 (S), 103 (Eta), 9 (h), 0.887 +/- 0.032 (Hd), 25.789 (k) and 17.980 x 10-3 +/- 0.003 (Pi), respectively (P < 0,05). The G + C content above 1434 sites positions of nucleotide sequences accounted for 0.542. The phylogenetic tree showed that these strains are divided into six groups. As observed, the appearance of isolated Vibrio on 3 organs of fish (S. ocellatus) hemorrhagic are V. azureus (27,67%), V. alginolyticus (50%), V. orientalis (6,67%) and V. fluvialis (16,67%). Through this result, we found that the diversity of Vibrio species that appeared on the red drum was used in the 16S rRNA region and the presence of toxin genes in these Vibrio species.

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