Virulence properties and pathogenicity of multidrug-resistant Vibrio harveyi associated with luminescent vibriosis in Pacific white shrimp, Penaeus vannamei

Wan Wan Ibrahim Nurhafizah; Kok Leong Lee; Abdul Razzak Laith A; Musa Nadirah; Muhd Danish-Daniel; Sandra Catherine Zainathan; Musa Najiah

doi:10.1016/j.jip.2021.107594

Virulence properties and pathogenicity of multidrug-resistant Vibrio harveyi associated with luminescent vibriosis in Pacific white shrimp, Penaeus vannamei

J Invertebr Pathol. 2021 Nov:186:107594. doi: 10.1016/j.jip.2021.107594. Epub 2021 Apr 18.

Authors

Wan Wan Ibrahim Nurhafizah¹, Kok Leong Lee¹, Abdul Razzak Laith A¹, Musa Nadirah¹, Muhd Danish-Daniel², Sandra Catherine Zainathan¹, Musa Najiah³

Affiliations

¹ Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia.
² Institute of Marine Biotechnology, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia.
³ Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia. Electronic address: najiah@umt.edu.my.

PMID: 33878330
DOI: 10.1016/j.jip.2021.107594

Abstract

Global high demand for Pacific white shrimp Penaeus vannamei has led to intensified cultivation and a wide range of disease problems, including bacterial diseases due to vibrios. Three presumptive luminescent Vibrio harveyi strains (Vh5, Vh8 and Vh10) were isolated from the hepatopancreas (Vh5) and haemolymph (Vh8 and Vh10) of diseased growout Pacific white shrimp from a farm in Setiu, Terengganu, Malaysia, using Vibrio harveyi agar (VHA) differential medium. All three strains were identified as V. harveyi by biochemical characteristics. 16S rRNA gene-based phylogenetic analyses by neighbour-joining, maximum likelihood and maximum parsimony methods showed all three strains in the V. harveyi cluster. All three strains were β-haemolytic and positive for motility, biofilm formation and extracellular products (caseinase, gelatinase, lipase, DNase, amylase and chitinase). Vh10 was subjected to pathogenicity test in Pacific white shrimp by immersion challenge and determined to have a LC₅₀ of 6.0 × 10⁸ CFU mL^-1 after 168 h of exposure. Antibiotic susceptibility tests showed that all strains were resistant to oxytetracycline (OXT30), oleandomycin (OL15), amoxicillin (AML25), ampicillin (AMP10) and colistin sulphate (CT25) but sensitive to doxycycline (DO30), flumequine (UB30), oxolinic acid (OA2), chloramphenicol (C30), florfenicol (FFC30), nitrofurantoin (F5) and fosfomycin (FOS50). Each strain was also resistant to a slightly different combination of eight other antibiotics, with an overall multiple antibiotic resistance (MAR) index of 0.40, suggesting prior history of heavy exposure to the antibiotics. Vh10 infection resulted in pale or discoloured hepatopancreas, empty guts, reddening, necrosis and luminescence of uropods, as well as melanised lesions in tail muscle. Histopathological examination showed necrosis of intertubular connective tissue and tubule, sloughing of epithelial cells in hepatopancreatic tubule, haemocytic infiltration, massive vacuolation and loss of hepatopancreatic tubule structure.

Keywords: Antibiotic susceptibility; Histopathological analysis; Luminescent Vibrio harveyi; MAR index; Pathogenicity; Penaeus vannamei.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Drug Resistance, Multiple, Bacterial*
Penaeidae / microbiology*
Vibrio / pathogenicity*
Vibrio / physiology*
Virulence

Supplementary concepts

Vibrio harveyi