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从尼罗罗非鱼肠道中分离潜在益生菌种以构建表达CC趋化因子的重组益生菌及其对尼罗罗非鱼先天免疫反应的有效性。

Isolation of Potential Probiotic spp. from the Intestine of Nile Tilapia to Construct Recombinant Probiotic Expressing CC Chemokine and Its Effectiveness on Innate Immune Responses in Nile Tilapia.

作者信息

Nakharuthai Chatsirin, Boonanuntanasarn Surintorn, Kaewda Jirawadee, Manassila Pimpisut

机构信息

School of Animal Technology and Innovation, Institute of Agricultural Technology, Suranaree University of Technology, 111 University Avenue, Muang, Nakhon Ratchasima 30000, Thailand.

出版信息

Animals (Basel). 2023 Mar 8;13(6):986. doi: 10.3390/ani13060986.

DOI:10.3390/ani13060986
PMID:36978530
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10044694/
Abstract

This study aimed to investigate the potential probiotic spp. from the intestine of Nile tilapia in order to construct a recombinant probiotic for the enhancement of the Nile tilapia immune response. One hundred bacterial isolates from the intestine of Nile tilapia were characterized for species identification using the 16s ribosomal RNA (rRNA). Only isolates with exhibited antagonistic activity were investigated for their biological functions, which included protease-producing capacity, bile salts and pH tolerance, antibiotic susceptibility, and pathogenicity tests. According to the best results, isolate B29, as closely related to , was selected to construct a recombinant probiotic for the delivery of CC chemokine protein (pBES-CC). The existence of recombinant probiotics was confirmed by Western blotting before the feeding trial. In addition, the CC chemokine mRNA level was quantified in the intestine of fish fed probiotics after 30 days of feeding. Total immunoglobulin, lysozyme activity, alternative complement 50 activity (ACH50), and phagocytic activity of fish fed either wild-type or recombinant probiotics were significantly increased, indicating that probiotics could stimulate the Nile tilapia immune system through different processes. Interestingly, the dietary supplementation of recombinant probiotics has a stronger immune response enhancement than the wild-type strain.

摘要

本研究旨在从尼罗罗非鱼肠道中筛选潜在的益生菌种，以构建重组益生菌来增强尼罗罗非鱼的免疫反应。利用16S核糖体RNA（rRNA）对从尼罗罗非鱼肠道中分离出的100株细菌进行菌种鉴定。仅对表现出拮抗活性的分离株进行生物学功能研究，包括产蛋白酶能力、胆汁盐和pH耐受性、抗生素敏感性及致病性测试。根据最佳结果，选择与[具体菌种名称未给出]密切相关的分离株B29构建用于递送CC趋化因子蛋白（pBES-CC）的重组益生菌。在饲喂试验前通过蛋白质免疫印迹法确认重组益生菌的存在。此外，在饲喂30天后对饲喂益生菌的鱼肠道中CC趋化因子mRNA水平进行定量分析。饲喂野生型或重组益生菌的鱼的总免疫球蛋白、溶菌酶活性、替代补体50活性（ACH50）和吞噬活性均显著增加，表明益生菌可通过不同过程刺激尼罗罗非鱼免疫系统。有趣的是，日粮中添加重组益生菌比野生型菌株具有更强的免疫反应增强作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/183dc292e040/animals-13-00986-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/b8cdb051e722/animals-13-00986-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/97904bec90e1/animals-13-00986-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/366ec9d397c0/animals-13-00986-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/fcc1baf29a64/animals-13-00986-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/10b5353d209e/animals-13-00986-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/6fa50add1778/animals-13-00986-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/0ab59eda1410/animals-13-00986-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/183dc292e040/animals-13-00986-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/b8cdb051e722/animals-13-00986-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/97904bec90e1/animals-13-00986-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/366ec9d397c0/animals-13-00986-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/fcc1baf29a64/animals-13-00986-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/10b5353d209e/animals-13-00986-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/6fa50add1778/animals-13-00986-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/0ab59eda1410/animals-13-00986-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8e3/10044694/183dc292e040/animals-13-00986-g008.jpg

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