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研究生: 伊庭蝶
Prabawati Estuningdyah
論文名稱: 一種含杏鮑菇廢棄物萃取物及植物乳酸菌之益合素飼料改善白蝦成長、腸道微生物菌相及健康
A synbiotic prepared from king oyster mushroom by-product and lactic acid bacterium improves the growth and health of white shrimp, Litopenaeus vannamei
指導教授: 劉俊宏
Chun-Hung Liu
胡紹揚
Shao-Yang Hu
學位類別: 碩士
Master
系所名稱: 農學院 - 生物科技系
Department of Biological Science and Technology
畢業學年度: 110
語文別: 英文
論文頁數: 93
中文關鍵詞: 白蝦杏鮑菇副產物生長性能免疫反應腸道微生物菌相
外文關鍵詞: king oyster mushroom by-product, intestinal microbial community
DOI URL: http://doi.org/10.6346/NPUST202200074
相關次數: 點閱:35下載:11
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  • 本研究評估了含有杏鮑菇副產品萃取物 (KOME) 和Lactobacillus plantarum 7-40 的益合素飼料對南美白對蝦生長、免疫反應和腸道菌群的影響。 KOME 顯著刺激了益生菌的生長,但不刺激弧菌病原體的生長,包括Vibrio alginolyticus, V. parahaemolyticus, 和V. harveyi。配製四種飼料,包括魚粉基礎飼料(control)、添加KOME(5 g kg-1)的基礎飼料(ME)、添加益生菌的基礎飼料(1×108 CFU kg-1)(LP),以及添加 KOME (5 g kg-1) 和益生菌 (1×108 CFU kg-1) (SYN) 的基礎飼料。攝食 ME、LP 和 SYN 飼料的白蝦存活率顯著高於對照組。在SYN組中,蝦的增重和最終總重明顯高於對照組。 SYN 組的蝦腸中的乳酸菌數量顯著增加,而 ME 組的弧菌數量顯著高於對照組。分析總體基因體檢測到 13 個門、21 個綱、51 個目、74 個科和 104 個屬。雖然沒有發現顯著差異,但SYN組表現出比對照組更高的豐富度、均勻度和辛普森多樣性指數。文氏圖數據顯示,對照組和 SYN 組共享 57 個操作分類單元 (OTU),但僅在對照組中發現了額外的細菌病原體。然而,與對照組相比,SYN 組顯著增加了腸道中乳酸桿菌Lactobacillus, Shewanella和 Hypnocyclicus 的積累。由於增加的免疫反應,包括酚氧化酶 (PO)、超氧化物歧化酶 (SOD)、溶菌酶活性 (LYZ) 和吞噬活性 (PA),餵食 SYN 的蝦對V. alginolyticus具有顯著抗性。此外,與對照組相比,給予益合素飼料後,血細胞中 pexn 和 pen4 基因以及肝胰腺中 lgbp、sp、propoii、pexn、pen3a、pen4 和 gpx 的基因表達顯著增加。因此,本研究的結果表明,將 KOME 和益生菌作為益合素飼料使用可改善白蝦的生長、健康及病原體抗性,並改變白蝦的腸道微生物菌相。

    This study evaluated the effects of a synbiotic diet containing the by-product of king oyster mushroom, Pleurotus eryngii (KOME), and the probiotic Lactobacillus plantarum 7-40 on growth performance, immune response, and intestinal microflora of Litopenaeus vannamei. KOME significantly stimulated the growth of probiotics, but not the growth of Vibrio-like pathogens, including V. alginolyticus, V. parahaemolyticus, and V. harveyi. Four feeds were formulated, including a fish meal basal diet (control), a basal diet supplemented with KOME (5 g kg-1) (ME), a basal diet supplemented with probiotic (1×108 CFU kg-1) (LP), and a basal diet supplemented with KOME (5 g kg-1) and probiotic (1×108 CFU kg-1) (SYN). Shrimp-fed ME, LP, and SYN diets showed significantly high survival rates than controls. In the SYN treatment, shrimp had significantly higher weight gain and total final weight than the control treatment. The number of lactic acid bacteria in the shrimp intestine was substantially higher in the SYN treatment, whereas the Vibrio-like bacteria count was significantly higher in the ME treatment than the control. Metagenomic analysis data detected 13 phyla, 21 classes, 51 orders, 74 families, and 104 genera. Although no significant difference was found, the SYN group showed a higher richness, evenness, Shannon and Simpson diversity index than the control group. Venn diagram data showed that 57 of the operational taxonomic units (OTUs) were shared between the control and SYN groups, but additional bacterial pathogens were only found in the control group. However, the SYN group significantly increased the accumulation of Lactobacillus, Shewanella, and Hypnocyclicus in the intestine compared to the control treatment. Shrimp fed SYN had significant resistance to V. alginolyticus due to increased immune responses, including phenoloxidase (PO), superoxide dismutase (SOD), lysozyme activity (LYZ), and phagocytic activity (PA). In addition, gene expression of pexn and pen4 genes in hemocyte and lgbp, sp, propoii, pexn, pen3a, pen4, and gpx in hepatopancreas was significantly increased after administration of synbiotic feed when compared to controls. Therefore, the results of this study suggest that the combined use of KOME and probiotics as a synbiotic diet improved the growth performance, health status, pathogen resistance and positively altered the intestinal microbial community of white shrimp and concurrently contributes to a more sustainable aquaculture practice.

    摘 要 I
    Abstract II
    Acknowledgments IV
    Table of Contents VI
    List of Tables VIII
    List of Figures IX
    1. PREFACE 1
    1.1 Introduction 1
    1.2 The objectives of the study 3
    2. LITERATURE REVIEW 5
    2.1 Prebiotic concept 5
    2.2 Mushroom as potential prebiotics 6
    2.2.1 Mushroom general characteristic and bioactive compounds 6
    2.2.2 King oyster mushroom characteristics and benefits 8
    2.2.3 Mushroom as potential prebiotics 9
    2.3 Probiotic concept 13
    2.4 Lac. plantarum as probiotic agent 14
    2.5 Synbiotic application 19
    2.5.1 Synbiotic concept 19
    2.5.2 Synbiotic in aquaculture 19
    2.6 White shrimp (L. vannamei) immune system 23
    3. MATERIAL AND METHODS 25
    3.1 King oyster mushroom water extract (KOME) preparation 25
    3.2 Probiotic preparation 25
    3.3 Experimental design 25
    3.4 Bacteria growth stimulated by KOME 26
    3.5 Growth Trial 27
    3.5.1 Experimental diet preparation 27
    3.5.2 Shrimp rearing 30
    3.6 Intestinal microflora counts 30
    3.7 Health status assessment 31
    3.7.1 Challenge test 31
    3.7.2 Immune response 32
    3.7.3 Immune-related gene expression analysis 35
    3.8 16S metagenomic microbial analysis 39
    3.8.1 Sample collection and DNA extraction 39
    3.8.2 16S DNA amplicon sequencing and data processing 39
    3.9 Statistical analysis 40
    4. RESULTS 41
    4.1. Bacteria growth stimulated by KOME 41
    4.2 Shrimp growth performance 41
    4.3 Intestinal microbiological analysis 44
    4.4 Health status analysis 44
    4.4.1 Challenge test 44
    4.4.2 Immune parameters 47
    4.4.3 Immune gene expression analysis 47
    4.5 Shrimp gut microbiome characteristic 53
    4.6 Intestinal microbiota similarities and differences 53
    4.7 Intestinal microbiome structure and diversity 56
    5. DISCUSSION 62
    6. CONCLUSION 69
    REFERENCES 70

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