Abstract

As a kind of antimicrobial peptides (AMP) in crustacean, anti-lipopolysaccharide factors (ALFs) have broad spectrum antimicrobial activities. In the present study, we identified four ALF genes, EcALF2-5, from the ridgetail prawn Exopalaemon carinicauda. Tissue distribution analysis showed that EcALF2 and EcALF4 transcripts were mainly located in gill, epidermis, and stomach, while EcALF3 and EcALF5 were mainly in hemocytes. Peptides corresponding to the LPS binding domain (LBD) of EcALFs were synthesized for analyzing their antimicrobial activities. Minimal inhibitory concentration (MIC) analysis showed that the synthetic LBD peptides of EcALF3 and EcALF4 could inhibit the growth of Gram-positive and Gram-negative bacteria, while the synthetic LBD peptides of EcALF2 and EcALF5 showed antibacterial activity against Vibrio. Incubation of white spot syndrome virus (WSSV) with the synthetic LBD peptides of EcALF3, EcALF4, and EcALF5 could reduce the in vivo viral copy number in WSSV-infected prawns. After silencing of EcALFs, Vibrio exhibited a rapid proliferation in the hepatopancreas of the prawn. The present data showed the important function of different EcALFs in modulating the in vivo bacterial and viral propagation in E. carinicauda. This study will provide new clues into the disease control in aquaculture.

Highlights

  • Antimicrobial peptides (AMPs) are a kind of immune effectors in the innate immunity of the living organisms

  • Since the first AMP named cecropin was isolated from the pupae of the cecropia moth Hyalophora cecropia [1], thousands of AMPs have been found in various species to date, such as defensins and cathelicidins in mammals [2], thionins and lipid transfer proteins (LTPs) in plants [3], attacins and lebocins in insects [4]

  • Shrimp to bacterial infection [22,23,24]. All these studies revealed that anti-lipopolysaccharide factors (ALFs) played important roles in Various isoforms of ALFs co-existing in one species exhibited different antimicrobial activities

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Summary

Introduction

Antimicrobial peptides (AMPs) are a kind of immune effectors in the innate immunity of the living organisms. The LPS binding domain (LBD), as the functional domain of ALF, contains two cysteine residues that can form a disulfide bond to stabilize the beta sheet structure of the domain [7,8,9]. ALF proteins certain antibacterial activities or inhibition to the propagation shrimp infected. ALFs played important roles in crustacean mortality of shrimp infected with Vibrio harveyi [21]. Shrimp to bacterial infection [22,23,24] All these studies revealed that ALFs played important roles in Various isoforms of ALFs co-existing in one species exhibited different antimicrobial activities. ALFpresent isoforms were isolated from a clues for understanding the role of ALFs in crustacean transcriptome data and their functions were analyzed. The present data will provide new clues for understanding the role of ALFs in crustacean immunity

Sequences Characters and Phylogenic Analysis of EcALFs
Tissue
Minimal
Antiviral of EcLBD
Optimization
Hemolytic Property of EcLBD Peptides sequences
Discussions
Animals and Tissue Sampling
Sequence Analysis of EcALFs
Total RNA Extraction and RT-qPCR Analysis
Synthesis of EcLBD Peptides
Antiviral Activity of EcLBDs
DsRNA Synthesis and Gene Silencing of EcALFs
Hemolytic Activity of EcLBDs
Statistical Analyses

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