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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/30161

Title: CBM 複合益生菌應用在水質改善及提升白蝦(Litopenaeus vannamei)免疫力
Application of Complex Beneficial Microorganisms (CBM) in water quality improvement and immunity enhancement of white shrimp (Litopenaeus vannamei)
Authors: Pei-Hsin Huang
黃姵馨
Contributors: NTOU:Department of Aquaculture
國立臺灣海洋大學:水產養殖學系
Keywords: 益生菌;水質;白蝦免疫
probiotic;water quality;white shrimp immunity
Date: 2011
Issue Date: 2011-11-25T07:31:43Z
Abstract: 益生菌是指對宿主有益的微生物和其產物,應用在水產養殖上,直接影響產量、物質循環、動物營養、水質、疾病控制。過去在水產養殖上大都以光合菌與硝化菌作為水質與底質改善劑,但效果有限。本研究之目標為將複合益生菌應用在水產養殖上,為達到此一目標,本論文進行了下列研究;(1)以即時定量聚合酶連鎖反應偵測定量複合型益生菌組合中各類微生物的含量;(2)應用複合型益生菌處理養殖廢水改善水質;(3)以複合型益生菌作為直接投餵微生物增加 白 蝦 免 疫 力 與 抗 病 能 力 。 由 於 CBM ( Complex Beneficial Microorganisms) 菌是一複合益生菌配方,有必要知道特定菌株在共棲培養過程中各類菌數的變化,本研究利用Real-time PCR對CBM菌進行定量,其中R. palustris, R. rubrum, R. sphaeroides, L. pentosus, L. lactis, Bacillus, S. fibuligera, O. anthropi, T. pantotropha, A. denitrificans, P. citrinum, C. globosum, A. oryzae, M. verrucaria, A. niger在共棲培養階段菌量分別為4.10 × 10 9 , 2.56 × 10 9 , 1.39× 10 8 , 3.57 × 10 8 , 7.90 × 10 8 , 1.42 × 10 8 , 2.13 × 10 8 , 2.75 × 10 8 , 4.69 × 10 8 , 5.22 × 10 8 , 3.85 × 10 8 CFU‧mL -1 ; 第一次擴量培養中,菌量分別為1.58 × 10 7 ~ 7.94 × 10 8 , 3.027 × 10 6 ~ 3.92 × 10 8 , 7.6 × 10 6 ~ 6.4 × 10 8 , 9.91 × 10 6 ~ 5. 16 × 10 8 , 9.91 × 10 6 ~ 6.77 × 10 8 , 3.55 × 10 6 ~ 7.36 × 10, 4.89 × 10 6 ~ 4.23 × 10 8 , 8.9 × 10 6 ~ 1.40 × 10 8 , 5.57 × 10 6 ~ 11.40 × 10 8 , 1.16 × 10 6 ~ 3.04 × 10 8 , 1.6 × 10 6 ~ 5.08 × 10 8 CFU‧mL -1 ; 第二次擴量培養後的菌量分別為0.58 × 10 7 ~ 7.94 × 10 8 , 3.02 × 10 6 ~ 3.92 × 10 8 , 1.6 × 10 6 ~ 2.4 × 10 7 , 3.12 × 10 6 ~ 1.41 × 10 7 , 3.12 × 10 5 ~ 1.41 × 10 7 , 1.84 × 10 6 ~ 7.5 × 10 7 , 1.30 × 10 6 ~ 4.17 × 10 7 , 4.67 × 10 6 ~ 2.76 × 10 7 , 6.1× 10 5 ~ 2.9 × 10 7 , 6.1× 10 5 ~ 3.75 × 10 7 , 1.04× 10 6 ~ 2.29 × 10 7 CFU‧mL -1。在水質處理試驗中,添加 100 ppm CBM菌 可改善淡水養殖廢水、海水養殖廢水、工業廢水中的氨氮、亞硝酸含量;在廢水中額外添加碳源可以增加 CBM 菌促進反硝化作用及廢水中氨氮及亞硝酸的降解能力。以直接投餵(Direct-Fed-Microbial)方式於南美白對蝦 (Litopenaeus vannamei)飼料裡添加 CBM 菌結果顯示 haemocyanin 、 crustin 及 penaeidin 等先天免疫基因表現量均有增加趨勢,其中haemocyanin、crustin mRNA表現量高出對照組兩倍以上。以CBM菌刺激白蝦免疫反應後以病源菌株及活化白點病毒株進行攻毒試驗中,結果顯示在哈威弧菌攻毒實驗中投餵CBM菌處理組白蝦可提高存活率約30﹪;在白點病毒攻毒實驗中投餵CBM菌處理組可提高白蝦存活率約 20﹪。
Probiotics, which are micro-organisms and their products with health benefit to the host, have used in aquaculture as a means of disease control, supplementing or even in some cases replacing the use of antimicrobial compounds. Water quality is the most important essentials in aquacluture industry. In past, it has been shown most probiotics can for improving water quality, are phototrophic bacteria and nitrifying bacteria. The objective of present study is to stablish quantitation method to measure specific bacteria in complex microorganisms and applications of Complex Beneficial Microorganisms (CBM) in water quality improvement and immunity enhancement in white shrimp(Litopenaeus vannamei). To accomplish this objective, several specific aims were established ; 1)apply real-time quantitative polymerase chain reaction (Real-Time Quantitative PCR, Q-PCR) to quantitate composite probiotic of CBM which contaning various of strains of microorganisms, 2)apply of CBM in simulated wastewater treatment for improving water quality, 3)use CBM as a direct-fed-microbial to enhance innate immunity and disease resistance in white shrimp. In present study, Q-PCR quantification of CBM bacteria, R. palustris, R. rubrum, R. sphaeroides,L. pentosus, L. lactis, Bacillus, S. fibuligera, O. anthropi, T. pantotropha, A. denitrificans, P. citrinum, C. globosum, A. oryzae, M. verrucaria, A.niger in symbiotic amplification culture process were; 4.10 × 10 9 , 2.56 × 10 9 , 1.39× 10 8 , 3.57 × 10 8 , 7.90 × 10 8 , 1.42 × 10 8 , 2.13 × 10 8 , 2.75 × 10 8 ,4.69 × 10 8 , 5.22 × 10 8 , 3.85 × 10 8 CFU‧mL -1 respectively; the cells number in first molasses amplification culture process were; 1.58 × 10 7~ 7.94 × 10 8 , 3.027 × 10 6 ~ 3.92 × 10 8 , 7.6 × 10 6 ~ 6.4 × 10 8 , 9.91 × 10 6 ~ 5. 16 × 10 8 , 9.91 × 10 6 ~ 6.77 × 10 8 , 3.55 × 10 6 ~ 7.36 × 10, 4.89 × 10 6 ~ 4.23 × 10 8 , 8.9 × 10 6 ~ 1.40 × 10 8 , 5.57 × 10 6 ~ 11.40 × 10 8 , 1.16 × 10 6~ 3.04 × 10 8 , 1.6 × 10 6 ~ 5.08 × 10 8 CFU‧mL-1 respectively; the cell numbers in second molasses amplification culture process were; 0.58 × 10 7 ~ 7.94 × 10 8 , 3.02 × 10 6 ~ 3.92 × 10 8 , 1.6 × 10 6 ~ 2.4 × 10 7 , 3.12 × 10 6 ~ 1.41 × 10 7 , 3.12 × 10 5 ~ 1.41 × 10 7 , 1.84 × 10 6 ~ 7.5 × 10 7 , 1.30 × 10 6 ~ 4.17 × 10 7 , 4.67 × 10 6 ~ 2.76 × 10 7 , 6.1× 10 5 ~ 2.9 × 10 7 , 6.1× 10 5 ~ 3.75 × 10 7 , 1.04× 10 6 ~ 2.29 × 10 7 CFU‧mL -1 respectively. In water quantity treatment tests, adding 100 ppm into freshwater aquaculture wastewater, marine wastewater, and industrial wastewater, had shown that the ammonia and nitrite levels reducing significantly. Extra carbon source additation can enhance denitrification process and reducing ammonia and nitrite level significantly. Using CBM as DFM experiment in white shrimp, the result indicated that the expression of haemocyanin, crustin and penaeidin genes increased, the mRNA levels of haemocyanin, crustin were twice amount than the control group. After CBM administration, the results of pathogens challenged test demonstrated that shrimp fed with survival rate CBM can be improved about 30% after Vibrio harveyi infection. The group fed with CBM had 20% higher survival rate after WSSV infection.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M98330023
http://ntour.ntou.edu.tw/handle/987654321/30161
Appears in Collections:[水產養殖學系] 博碩士論文

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