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Research article2019Peer reviewedOpen access

PirAB(VP) Toxin Binds to Epithelial Cells of the Digestive Tract and Produce Pathognomonic AHPND Lesions in Germ-Free Brine Shrimp

Kumar, Vikash; De Bels, Lobke; Couck, Liesbeth; Baruah, Kartik; Bossier, Peter; Van den Broeck, Wim

Abstract

Acute hepatopancreatic necrosis disease (AHPND), a newly emergent farmed penaeid shrimp bacterial disease originally known as early mortality syndrome (EMS), is causing havoc in the shrimp industry. The causative agent of AHPND was found to be a specific strain of bacteria, e.g., Vibrio and Shewanella sps., that contains pVA1 plasmid (63-70 kb) encoding the binary PirA(VP) and PirB(VP) toxins. The PirAB(VP) and toxins are the primary virulence factors of AHPND-causing bacteria that mediates AHPND and mortality in shrimp. Hence, in this study using a germ-free brine shrimp model system, we evaluated the PirAB(VP) toxin-mediated infection process at cellular level, including toxin attachment and subsequent toxin-induced damage to the digestive tract. The results showed that, PirAB(VP) toxin binds to epithelial cells of the digestive tract of brine shrimp larvae and produces characteristic symptoms of AHPND. In the PirAB(VP)-challenged brine shrimp larvae, shedding or sloughing of enterocytes in the midgut and hindgut regions was regularly visualized, and the intestinal lumen was filled with moderately electron-dense cells of variable shapes and sizes. In addition, the observed cellular debris in the intestinal lumen of the digestive tract was found to be of epithelial cell origin. The detailed morphology of the digestive tract demonstrates further that the PirAB(VP) toxin challenge produces focal to extensive necrosis and damages epithelial cells in the midgut and hindgut regions, resulting in pyknosis, cell vacuolisation, and mitochondrial and rough endoplasmic reticulum (RER) damage to different degrees. Taken together, our study provides substantial evidence that PirAB(VP) toxins bind to the digestive tract of brine shrimp larvae and seem to be responsible for generating characteristic AHPND lesions and damaging enterocytes in the midgut and hindgut regions.

Keywords

PirAB(VP) toxins; brine shrimp; digestive tract; sloughing; epithelial cells; necrosis

Published in

Toxins
2019, Volume: 11, number: 12, article number: 717
Publisher: MDPI

    UKÄ Subject classification

    Fish and Aquacultural Science

    Publication identifier

    DOI: https://doi.org/10.3390/toxins11120717

    Permanent link to this page (URI)

    https://res.slu.se/id/publ/104031