Hepcidin-Expressing Fish Eggs as A Novel Food Supplement to Modulate Immunity against Pathogenic Infection in Zebrafish ( Danio rerio )

Chung-Chih Tseng, Tah-Wei Chu, Ridha Handriany Danata, Yenny Risjani, Hui-Tsu Shih and Shao-Yang Hu
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Chung-Chih Tseng: Department of Dentistry, Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung 81357, Taiwan
Tah-Wei Chu: Department and Graduate Institute of Aquaculture, National Kaohsiung University of Science and Technology, Kaohsiung 811, Taiwan
Ridha Handriany Danata: Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan
Yenny Risjani: Department of Fisheries and Marine Science, University of Brawijaya, Malang 65145, Indonesia
Hui-Tsu Shih: Department of Clinical Pharmacy, Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung 81357, Taiwan
Shao-Yang Hu: Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan

Sustainability, 2020, vol. 12, issue 10, 1-15

Abstract: Hepcidin antimicrobial peptides are difficult to produce in prokaryotic expression systems due to their complex structure and antimicrobial activity. Although synthetic hepcidin provides an alternative to solve this issue, its high cost limits its practical application in various industries. The present study used zebrafish eggs as bioreactors to produce convict cichlid ( Amatitlania nigrofasciata ) hepcidin (AN-hepc) using the oocyte-specific zona pellucida ( zp3 ) promoter. The expression plasmid pT2-ZP3-AN-hepc-ZP3-EGFP, using EGFP as a reporter of AN-hepc expression, was designed to establish the transgenic line Tg(ZP3:AN-hepc:ZP3:EGFP) for the expression of AN-hepc. The AN-hepc peptide was produced successfully in fertilized eggs, as evidenced by RT-PCR and Western blotting. The AN-hepc-expressing eggs exhibited antimicrobial activity against a variety of aquatic pathogens and antibiotic-resistant pathogens, suggesting that the AN-hepc expressed in fish eggs was bioactive. The immunomodulatory effects of AN-hepc-expressing fertilized eggs on zebrafish innate immunity were evaluated by determining the expression of indicator genes after feeding with AN-hepc-expressing fertilized eggs for two months. Zebrafish supplementation with AN-hepc-expressing fertilized eggs significantly increased the expression of innate immunity-related genes, including IL-1β, IL-6, IL-15, TNF-α, NF-κb, complement C3b, lysozyme and TLR-4a. The zebrafish administered AN-hepc-expressing eggs exhibited higher cumulative survival than fish supplemented with wild-type and control eggs after infection with Aeromonas hydrophila and Streptococcus iniae . In conclusion, the present results showed that supplementation with AN-hepc-expressing fish eggs enhanced zebrafish innate immunity against pathogenic infections, suggesting that fertilized eggs containing AN-hepc have the potential to be developed as a food supplement for improving health status in aquaculture.

Keywords: hepcidin; bioreactor; innate immunity; disease resistance; zebrafish (search for similar items in EconPapers)
JEL-codes: O13 Q Q0 Q2 Q3 Q5 Q56 (search for similar items in EconPapers)
Date: 2020
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