Screening and Characterization of Potential Antioxidant Probiotics Isolated from the Gut of Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)
Screening and Characterization of Potential Antioxidant Probiotics Isolated from the Gut of Hybrid Grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)
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从混合石斑鱼（Epinephelus fuscoguttatus — — Epinephelus lanceolatus —）肠道中分离的潜在抗氧化益生菌的筛选和表征
DOI：
10.3389/fmars.2022.933304
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发表时间：
2022-07
影响因子：
3.7
通讯作者：
Shifeng Wang
中科院分区：
文献类型：
--
作者：
Qian Zhang;Yan Cai;Zihan Chen;Huizhong Shi;Yongcan Zhou;Yang Yang;Rui Tu;Guisen Chen;Shifeng Wang
一些共生益生菌具有抗氧化活性，可以提高宿主的抗氧化能力。目前还没有关于石斑鱼养殖中宿主源抗氧化益生菌筛选的报道。本研究从583株杂交石斑鱼肠道分离菌株中筛选出369株具有非溶血性的菌株。随后的初步筛选与2，2-二苯基-1-苦肼（DPPH）清除率测定产生36个潜在的抗氧化剂分离物。通过DPPH自由基清除率、2，2 '-连氮双（3-乙基苯并噻唑啉）-6-磺酸自由基清除率、铁离子还原能力、还原活性、O2- ·清除率、·OH清除率、亚铁离子螯合率、过氧化氢耐受性、氧自由基清除能力等9项抗氧化试验进行综合评价，从36株具有抗氧化活性的菌株中筛选出10株抗氧化能力较强的菌株。然后对这10株菌株的模拟胃肠液耐受性、粘附性、消化酶活性和抗菌活性等其他益生菌特性进行了评价。所有10个分离株也进行了鉴定，使用分子方法。最后，从10个分离物中选择红色弧菌GO 91和珊瑚希瓦氏菌GO 310作为从10个分离物鉴定得到的两个属的代表，并且具有最佳的总体益生菌特性。进一步检测了分离株GO 91和GO 310的安全性能。抗生素敏感性试验结果表明，GO 91和GO 310对多种常用水产抗生素敏感。GO 91和GO 310的体内攻毒试验均未引起杂鱼的发病症状和死亡。因此，GO 91和GO 310菌株具有作为益生菌应用于杂交石斑鱼养殖的潜力。
Some symbiotic probiotics have antioxidant activities and could improve the antioxidant capacity of the host. There is still no report on the screening of host-derived antioxidant probiotics for grouper farming. In this study, 369 out of 583 isolates were screened from the gut of hybrid grouper ( Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂ ) based on their non-hemolytic characteristics. Subsequent preliminary screening with 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) scavenging rate assay resulted in 36 potential antioxidant isolates. After comprehensive evaluation with nine different antioxidant assays (DPPH scavenging rate, 2,2’-azino-bis (3-ethylbezothiazoline)-6-sulfonic acid radical scavenging rate, iron ion reducing ability, reducing activity, O 2 - · scavenging rate,·OH scavenging rate, ferrous ion chelating rate, hydrogen peroxide tolerance, oxygen-free radical scavenging ability, et al.), 10 isolates with strong antioxidant abilities were screened from 36 potential antioxidant isolates. Then some other probiotic properties, such as simulated gastrointestinal fluid tolerance, adhesion, digestive enzyme activity, and antibacterial activity of the 10 selected isolates were evaluated. All 10 isolates were also identified using the molecular method. Finally, Vibrio rhodolitus GO 91 and Shewanella corallii GO 310, as representatives of the two genera resulting from the identification of the 10 isolates, and with the best overall probiotic properties, were selected from the 10 isolates. Isolates GO 91 and GO 310 were further tested for their safety performances. Antibiotic sensitivity tests showed that GO91 and GO310 were sensitive to many commonly used aquaculture antibiotics. The in vivo challenge test of GO 91 and GO 310 didn’t cause any disease symptoms or death in hybrid grouper. Therefore, isolates GO 91 and GO 310 showed great potential to be used as probiotics in hybrid grouper farming.
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