沙福芽孢桿菌 (Bacillus safensis) 對白蝦成長促進與免疫調節之研究

Effects of dietary Bacillus safensis on growth performance and immune modulation of white shrimp (Litopenaeus vannamei)

米俊仲(Mi Jun-Zhong)

指導教授 ： 胡紹揚

國立屏東科技大學/農學院/生物科技系/碩士(2021年)

https://doi.org/10.6346/NPUST202100403

摘要

本研究由吳郭魚腸道中篩選出一株具澱粉酶、蛋白酶、纖維素酶及木聚醣酶活性之菌株，經過16S rDNA序列鑑定及生化分析，確認該菌株為沙福芽孢桿菌 (Bacillus safenesis)。抗生素敏感性試驗結果顯示此菌株對於數種抗生素均無抗藥性。為評估其是否具有作為水產養殖益生菌之潛力，在自製飼料中添加105 CFU/g、106 CFU/g與107 CFU/g的B. safenesis，並餵食白蝦8週，分析該菌株對白蝦的生長促進、免疫調節、疾病抵抗及肝胰腺菌相之改善。結果顯示，添加106 CFU/g與107 CFU/g的組別，其體重增加及飼料效益方面皆顯著高於未添加B. safenesis的控制組。在免疫指標分析中，攝食107 CFU/g的B. safenesis的白蝦，其血液中酚氧化酶、超氧化物歧化酶、呼吸爆、溶菌酶及吞噬活性皆顯著高於控制組；另外，105 CFU/g及106 CFU/g組別在酚氧化酶、呼吸爆及吞噬活性分析中，亦有提升活性的趨勢。在免疫基因分析中，以real-time PCR分析免疫相關基因prophenoloxidase、TNF-α、astakine以及lysozyme的表現量，其中106 CFU/g與107 CFU/g的組別在四種基因表現量皆顯著高於控制組，105 CFU/g的組別在TNF-α、astakine及lysozyme的表現量相較於控制組亦有顯著提升。以溶藻弧菌 (Vibrio alginolyticus) 注射白蝦感染7天後，結果顯示餵食106 CFU/g與107 CFU/g的組別，其累積死亡率顯著低於控制組。次世代定序結果說明白蝦攝食B. safenesis有助於改變其菌相。綜上所述，每日以每克飼料含106 CFU之B. safenesis餵食白蝦，有助於白蝦之生長促進、免疫調節、疾病抵抗以及肝胰腺菌相之變化，說明B. safenesis具有潛力開發作為應用於水產養殖之益生菌。

關鍵字

南美白蝦 ； 益生菌 ； 沙福芽孢桿菌 ； 成長促進 ； 免疫調節

並列摘要

In the present study, a strain with amylase, protease, cellulase and xylanase activities, was isolated from the intestine of Nile tilapia and characterized as Bacillus safensis by 16S rDNA sequencing and biochemical analysis. In order to evaluate the efficacy of B. safensis as a probiotic on improvement of growth performance, immune modulation, disease resistance and gut microbiota of white shrimp (Litopenaeus vannamei), shrimp was fed with diet containing 105 CFU/g (G1), 106 CFU/g (G2) or 107 CFU/g (G3) for eight weeks. The results showed that weight gain and feed efficiency in the shrimps of G2 and G3 groups were significantly increased than those in shrimp of control group. Immune parameters including phenoloxidase, superoxide dismutase (SOD), respiratory burst, lysozyme and phagocytic activities in shrimp fed diet containing 107 CFU/g of B. safenesis were significantly higher than those in shrimp of control group. In addition, shrimps of G1 and G2 groups were also showed significant difference from the shrimp of control group in phenoloxidase, respiratory burst and phagocytic activity. The expression level of prophenoloxidase, TNF-α, astakine and lysozyme genes in hepatopancreas of shrimps in G1, G2 and G3 groups were significantly higher than those in shrimp of control group. A significant decreasing of cumulative mortality was exhibited in white shrimp fed diet containing 106 CFU/g and 107 CFU/g of B. safenesis after challenging with Vibrio alginolyticus. The shrimps dietary supplemented with B. safensis revealed an improved microbiota in hepatopancreas of white shrimp were performed by next generation sequencing (NGS). In summary, dietary feeding of diet containing 106 CFU/g of B. safenesis could improve growth performance, immune modulation, disease resistance and microbiota of white shrimp. The results suggested that B. safenesis has potential to be developed as a probiotic for aquaculture.

並列關鍵字

Litopenaeus vannamei ； probiotics ； Bacillus safensis ； growth performance ； immune modulation

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