Abstract

Simple SummaryThe application of chemicals and drugs in fish culture has increased the existence of antimicrobial-resistant bacteria and drug residues in fish. Developing an effective vaccine to reduce specific disease-related losses and avoid antibiotic use has become an increasingly important part of aquaculture. Our current study found that the oral inactivated V. harveyi strain VH1 vaccine confers high protection in fish post-infection with virulent Vibrio spp. under experimental conditions. The oral vaccine was further tested for field efficacy and showed a higher survival and growth performance in vaccinated fish than unvaccinated fish. This current vaccine was shown to potentially provide sufficient protection to the host in both controlled and field environments against vibriosis.Vibriosis is one of the most common threats to farmed grouper; thus, substantial efforts are underway to control the disease. This study presents an oral vaccination against multiple Vibrio spp. in a marine fish with double booster immunisation. The Vibrio harveyi strain VH1 vaccine candidate was selected from infected groupers Epinephelus sp. in a local farm and was formalin inactivated and combined with commercial feed at a 10% ratio (v/w). A laboratory vaccination trial was conducted for seventy days. The induction of IgM antibody responses in the serum of Asian seabass Lates calcarifer immunised with the oral Vibrio harveyi strain VH1 was significantly (p < 0.05) increased as early as week one post-primary vaccination. Subsequent administration of the first and second booster for 5 consecutive days, starting on days 14 and 42, respectively, improved the specific antibody level and reached a highly significant (p < 0.05) value at days 35 and 49 before slightly decreasing from day 56 onwards. Antibody titres of the control unvaccinated group remained relatively stable and low throughout the experimental period. At the end of the 70-day vaccination trial, 23 days post final boost, an intraperitoneal challenge with a field strain of Vibrio harveyi, V. alginolyticus, and V. parahaemolyticus was carried out. Our challenge study showed that oral Vibrio harveyi strain VH1 vaccine candidate could induce significant protection, with an RPS of 70–80% against different Vibrio species. Thereafter, a field trial was conducted in a mariculture farm to study the effect of field vaccination using the oral Vibrio harveyi strain VH1 vaccine candidate. A total of 3000 hybrid grouper juveniles were divided into two groups in triplicate. Fish of Group 1 were not vaccinated, while Group 2 were vaccinated with the feed-based vaccine. Vaccinations were carried out on days 0, 14, and 42 via feeding the fish with the vaccine at 4% body weight for 5 consecutive days. At the end of the study period, the fish survival rate was 80% for the vaccinated group, significantly (p < 0.05) higher than the 65% seen in the control unvaccinated group. Furthermore, the vaccinated fish showed significantly (p < 0.05) better growth performances. Therefore, the oral Vibrio vaccine from the inactivated Vibrio harveyi strain VH1 is a potential versatile vaccine candidate that could stimulate good immune responses and confer high protection in both Asian seabass, Lates calcarifer, and farm hybrid grouper Epinephelus fuscoguttatus × Epinephelus lanceolatus.

Highlights

  • In Malaysia and its neighbouring countries, with a year-round tropical climate of 28 ◦ C, vibriosis has been frequently recorded at many marine aquaculture farms

  • V. harveyi and V. parahaemolyticus increased significantly (p < 0.05) until day 35, when they reached a high value, while in V. alginolyticus, they increased until day 28, before slightly dropping until day 42

  • Following the second booster dose on day 42, the IgM levels against V. harveyi and V. alginolyticus increased significantly (p < 0.05) on day 49, while for V. parahaemolyticus, they increased on day

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Summary

Introduction

In Malaysia and its neighbouring countries, with a year-round tropical climate of 28 ◦ C, vibriosis has been frequently recorded at many marine aquaculture farms. Vibrio harveyi, V. parahaemolyticus, V. alginolyticus, and V. anguillarum were most frequently isolated, affecting Asian seabass Lates calcarifer, brown-marbled grouper. Mohamad et al [5] reported an outbreak in Selangor, Malaysia, where farmed juvenile hybrid grouper (E. polyphekadion × E. fuscoguttatus) experienced losses of 29% in 10 days, with diseased fish becoming lethargic, displaying excessive mucus production, fin rot, congestion of the brain, liver, and kidneys, and splenic enlargement. Vibrio harveyi and V. alginolyticus were successfully recovered from the diseased hybrid groupers and may cause the infection of the fish host. Another study by Amalina et al [6] on groupers obtained from nine farms located at different geographical regions in Malaysia had detected multiple Vibrio spp. from. The incidence creates an urgent need to develop versatile or combined vaccines to simultaneously control fish disease caused by multiple pathogens [8]

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