Abstract

Omega-3 polyunsaturated fatty acids (n-3 PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exhibit antibacterial and anti-inflammatory activities. Furthermore, diets rich in n-3 PUFAs are known to improve disease resistance and limit pathogen infection in commercial aquaculture fishes. In this study, we examined the effects of transgenic overexpression of n-3 PUFA biosynthesis genes on the physiological response to bacterial infection in tilapia. We first established tilapia strains with single or dual expression of salmon delta-5 desaturase and/or delta-6 desaturase and then challenged the fish with Vibrio vulnificus infection. Interestingly, our data suggest that n-3 PUFA-mediated alterations in gut microbiota may be important in determining disease outcome via effects on immune response of the host. Both liver- and muscle-specific single and dual expression of delta-5 desaturase and delta-6 desaturase resulted in higher n-3 PUFA content in transgenic fish fed with a LO basal diet. The enrichment of n-3 PUFAs in dual-transgenic fish is likely responsible for their improved survival rate and comparatively reduced expression of inflammation- and immune-associated genes after V. vulnificus infection. Gut microbiome analysis further revealed that dual-transgenic tilapia had high gut microbiota diversity, with low levels of inflammation-associated microbiota (i.e., Prevotellaceae). Thus, our findings indicate that dual expression of transgenic delta-5 and delta-6 desaturase in tilapia enhances disease resistance, an effect that is associated with increased levels of n-3 PUFAs and altered gut microbiota composition.

Highlights

  • The shared biosynthetic pathway of omega-3 polyunsaturated fatty acids (n-3 PUFAs) eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22: 6 n-3) utilizes α-linolenic acid (ALA, 18: 3 n-3) as a starting material and involves several desaturation and elongation steps that are performed by delta-4 desaturase, delta-5 desaturase, delta-6 desaturase, delta-5 elongase and delta-6 elongase [1]

  • The genes were under the control of a tetracycline responsive element (TRE) fused with CMV minimal (CMVmini) promoter, which is activated in liver or muscle by tTA transactivator expressed by the first vector

  • The second vector contains TcCFP13 or enhanced eTcRFP11 reporter genes that are bi-directionally expressed with delta-5 desaturase or delta-6 desaturase genes of Atlantic salmon, respectively; the genes are under the control of a TRE fused with CMV promoter, which is activated in liver or muscle by tTA transactivator expressed by the first vector (Fig 1A)

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Summary

Introduction

The shared biosynthetic pathway of omega-3 polyunsaturated fatty acids (n-3 PUFAs) eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22: 6 n-3) utilizes α-linolenic acid (ALA, 18: 3 n-3) as a starting material and involves several desaturation and elongation steps that are performed by delta-4 desaturase, delta-5 desaturase, delta-6 desaturase, delta-5 elongase and delta-6 elongase [1]. The delta-6 desaturase enzyme has been shown to be rate limiting for the conversion of ALA to EPA in the lipid biosynthesis pathway [3]. Once synthesized, these and other n-3 PUFAs may function to improve animal health, at least partially as a result of their antibacterial and anti-inflammatory effects. The ability to synthesize DHA and EPA may be highly beneficial to an organism’s defense against infection

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