Transgenic microalgae as a non-antibiotic bactericide producer to defend against bacterial pathogen infection in the fish digestive tract
Abstract
Antibiotics are commonly employed in most fish aquacultures to prevent disease. One major risk in this practice is that antibiotic-resistant pathogens may be selected. Therefore, we wanted to examine the feasibility of producing an economical, non-antibiotic alternative. The microalga Nannochloropsis oculata is an essential phytoplankton used as live feed for fish larvae. We attempted to culture N. oculata in a way that would provide an organism against bacterial pathogenic infection. To test this idea, we constructed an algae-codon-optimized bovine lactoferricin (LFB) fused with a red fluorescent protein (DsRed) driven by a heat-inducible promoter, which is a heat shock protein 70A promoter combined with a ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit 2' promoter from Chlamydomonas reinhardtii. After electroporation, we examined 491 microalgal clones and generated two stable transgenic lines, each expressing a stable transgene inheritance for at least 26 months. This was confirmed by the positive detection of the mRNA transcript and the protein of LFB-DsRed produced by the transgenic microalgae. To test the efficacy of the antimicrobial peptide LFB, medaka fish (Oryzias latipes) were adapted from freshwater to seawater and were fed with the transgenic algae by oral-in-tube delivery method. Bacterial infection with 1 x 10(5)Vibrio parahaemolyticus per fish was induced 6h thereafter by oral-in-tube delivery as well. For medaka fish fed with 1 x 10(8) transgenic algae per fish, the average survival rate after a 24-h period of infection was much higher than that of medaka fed with wild-type algae (85+/-7.1% versus 5+/-7.1%). This result suggests that medaka fish fed with the LFB-containing transgenic microalgae will have bactericidal defense against V. parahaemolyticus infection in its digestive tract.
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Expression of bovine lactoferricin was achieved in nuclear transformants of Nannochloropsis oculata, and algal extracts from strains expressing this protein were effective against Escherichia coli and Vibrio parahemolyticus.
“Recently, expression of bovine lactoferricin was achieved in nuclear transformants of Nannochloropsis oculata. Algal extracts from strains expressing this protein were effective against Escherichia coli and Vibrio parahemolyticus [82].”
From Biofuels from algae: Challenges and potential (2010) · cites this paper as [82]
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- Authors
- Li, S.-S., & Tsai, H.-J.
- Affiliations
- National Taiwan University (TW)
- Corresponding author
- Tsai, Huai-Jen
- ORCID
- Tsai, Huai-Jen
- Venue
- Fish & Shellfish Immunology
- Publisher
- Elsevier BV
- Published
- 2009-02
- Volume
- 26
- Issue
- 2
- Pages
- 316-325
- ISSN
- 1050-4648 (ISSN-L)
- OpenAlex
- W1977835296
- Cited by
- 130
- References
- 40
- Citations by year
- 2026: 3 · 2025: 4 · 2024: 8 · 2023: 6 · 2022: 5 · 2021: 10 · 2020: 11 · 2019: 9 · 2018: 3 · 2017: 6 · 2016: 9 · 2015: 14 · 2014: 10 · 2013: 9 · 2012: 12
- FWCI
- 1.39
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Fish & Shellfish Immunology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Council
- National Taiwan University
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Aquaculture disease management and microbiota
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Agricultural and Biological Sciences › Aquatic Science › Aquaculture Nutrition and Growth
- Keywords
- transgenic microalgae, Nannochloropsis oculata, bovine lactoferricin, antimicrobial peptides, Vibrio parahaemolyticus, fish bacterial disease, medaka, live feed, antibiotic alternatives, heat-inducible promoter, codon optimization, electroporation, fish farming
- MeSH terms
- Animals, Fish Diseases, Lactoferrin, Oryzias, Plasmids, Protoplasts, Recombinant Proteins, Vibrio Infections, Vibrio parahaemolyticus, Blotting, Western, Survival Analysis, Algal Proteins, Organisms, Genetically Modified, Gastrointestinal Tract, Eukaryota
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