臺灣博碩士論文加值系統

本論文探討飼料中添加胚芽乳酸桿菌 (Lactobacillus plantarum, LP) 及嗜酸乳酸桿菌(Lactobacillus acidophilus, LA)，其投餵白蝦 (Litopenaeus vannamei)對於水質、成長表現及非特異性免疫之影響。

實驗組別包括兩株乳酸菌及三種不同濃度 (10^4、10^6及10^8 cfu/g)，分別命名為LP4、LP6、LP8及LA4、LA6、LA8。觀察8週投餵結果顯示，兩種乳酸菌飼料均可有效降低水中總氨氮及亞硝酸－氮的累積，但對水中總菌數及弧菌數無控制效果。另外各組水中可檢測到乳酸菌，濃度介於1.1 × 10^1 - 2.46 × 10^2 cfu/ml之間。

白蝦成長表現方面，各實驗組皆可提升白蝦成長，其中以LP8組別最為顯著，擁有最佳的增重率 (70.81 ± 5.25 %)、特殊成長率 (0.96 ± 0.06 %)及飼料轉換率 (3.94 ± 0.38)。

探討非特異性免疫方面，於投餵第 0、1、4、7、14及28天進行觀察，結果顯示兩株乳酸菌皆可提升免疫能力。在投餵4天後，總血球數開始有顯著提升情形。酚氧化酵素活性提升在第4天可達到最高峰，其中LA8組有最高的酚氧化酵素活性。超氧陰離子產生率在第4天有最顯著提升，除了LP4組，其餘組別均顯著高於控制組 (p < 0.05)，其中LA8組有最高的超氧陰離子產生率。吞噬率在投餵第7天後可觀察到顯著提升情形，其中LA8組提升最顯著。吞噬指數第14天及第28天各處理組均顯著高於控制組 (p < 0.05)，其中LP8組在第14天有最高的吞噬指數。

本研究證實，飼料添加胚芽乳酸桿菌 (L. plantarum)及嗜酸乳酸桿菌 (L. acidophilus)，可有效提升白蝦成長表現與非特異性免疫反應能力。

The objective of this study was to investigate the effects of dietary Lactobacillus plantarum and Lactobacillus acidophilus on the water qualities, growth performance and non-specific immunity of white shrimp (Litopenaeus vannamei) aquaculture.

Two probiotics and three concentrations (10^4, 10^6, 10^8 cfu g-1) were included in the experiment diets and reffered as LP4, LP6, LP8 and LA4, LA6, LA8. During the 8 weeks of feeding period, dietary supplement with both Lactobacillus showed the abilities to reduce the accumulation of NH3-N and NO2-N, but not to control the Vibrio sp. and total bacteria counts. Moreover, the concentrations of Lactobacillus in aquatic water were detected in the range of 1.1 × 101 to 2.46 × 102 cfu/mL.

In viewing of the growth performance, it was shown that the growth of L. vannamei were improved in individuals fed with probiotics. The greatest improvements in growth performances were observed in group LP8, which has the best weight gain (70.81 ± 5.25 %), specific growth rate (0.96 ± 0.06 %) and feed conversion rate (3.94 ± 0.38).

The effects of dietary probiotics on non-specific immunity were evaluated on 0, 1, 4, 7, 14 and 28 days. The results showed that all treatment groups were significantly stimulated. The significant increases of THC (total haemocyte count) were found after day 4. The improvement of PO (phenoloxidase activity) reached the highest peak at day 4 and group LA8 showed the highest PO activity. The most significant improvement of O2- (superoxide anion production ratio) was observed at day 4 and group LA8 showed the highest. Except for group LP4, the O2- of treatment groups were significantly higher than that of control group at day 4 (p < 0.05). The significant increases of PR (phagocytic rate) were shown after day 7 and the increase of PR in group LA8 was the most significant. The PI (phagocytic index) of all treatment groups were significantly higher than that of control group at day 14 and 28 (p < 0.05). Group LP8 showed the highest PI at day 14.

This study demonstrated that dietary supplements with L. plantarum and L. acidophilus are effective for enhancing the growth and non-specific immune responses of white shrimp.

謝辭..................................................I
摘要.................................................II
Abstract............................................III
目錄.................................................IV
表目錄..............................................VIII
圖目錄..............................................VIII
第一章 前言...........................................1
第二章 文獻整理........................................3
2.1. 南美白對蝦.......................................3
2.1.1. 分類學位置.....................................3
2.1.2. 白蝦生態與養殖發展..............................3
2.2. 益生菌...........................................4
2.2.1. 何謂益生菌.....................................4
2.2.2. 水產益生菌定義..................................4
2.2.3. 益生菌抗病機制..................................5
2.3. 乳酸菌...........................................7
2.3.1.乳酸菌群........................................7
2.3.2.乳酸桿菌........................................8
2.3.3. 乳酸菌與腸胃道之關係............................8
2.3.4. 乳酸桿菌於水產養殖之應用.........................9
2.4. 甲殼類免疫機制....................................9
2.4.1. 體液性 (Humoral ) 免疫：.......................10
2.5.2. 細胞性 (Cellular ) 免疫：.....................11
第三章、材料與方法.....................................15
3.1. 益生菌之操作.....................................15
3.1.1. 菌株來源.......................................15
3.1.2. 培養基選擇.....................................15
3.1.3. 菌種保存.......................................15
3.2. 實驗一、益生菌飼料之製備及菌量變化.................16
3.2.1. 益生菌之接種液.................................16
3.2.2. 益生菌培養流程.................................16
3.2.3. 菌株吸光值與菌懸液所含活菌數之關係...............16
3.2.4. 益生菌飼料之製備流程............................16
3.2.5. 益生菌飼料菌量之確效............................16
3.3. 實驗二、飼料中添加益生菌對白蝦成長、水質、水中菌相菌之影響...................................................17
3.3.1. 白蝦來源......................................17
3.3.2. 實驗設計......................................17
3.3.3. 實驗方法......................................17
3.3.4. 飼料中添加乳酸菌對白蝦成長之影響.................18
3.3.5. 飼料中添加益生菌對白蝦水質改善之測試.............18
3.3.6. 飼料中添加益生菌對養殖池水中細菌之影響............19
3.4. 實驗三、飼料中添加乳酸菌對白蝦非特異性免疫之影響.....20
3.4.1. 白蝦來源.......................................20
3.4.2. 實驗流程圖.....................................20
3.4.3. 實驗方法.......................................21
3.4.4. 血球懸浮液製備..................................21
3.4.5. 酚氧化酵素 (phenoloxidase，PO)：...............21
3.4.6. 吞噬作用 (Phagocytic ratio , PR ; Phagocytic index , PI) 測定：.........................................21
3.4.7. 超氧陰離子 (superoxide anion，O2-) 之分析：....21
3.5. 統計分析.........................................22
第四章、結果..........................................23
4.1. 飼料中乳酸菌菌量變化..............................23
4.2. 飼料中添加益生菌對白蝦成長之影響...................23
4.3. 飼料中添加乳酸菌對白蝦養殖池水水質之影響............23
4.3.1. 總氨氮.........................................23
4.3.2. 亞硝酸-氮......................................23
4.4. 飼料中添加益生菌對白蝦養殖池水菌量之影響...........24
4.4.1. 總生菌數.......................................24
4.4.2. 弧菌數.........................................24
4.4.3. 乳酸菌數 (LAB).................................24
4.5. 飼料中添加乳酸菌對白蝦非特異性免疫之影響............24
4.5.1. 飼料添加不同乳酸菌對白蝦總血球數之影響............24
4.5.2. 飼料添加不同乳酸菌對白蝦酚氧化酵素 (PO)之影響.....25
4.5.3. 飼料添加不同乳酸菌對白蝦超氧陰離子 (O2-)產生率之影響 .....................................................25
4.5.4. 飼料添加不同乳酸菌對白蝦吞噬率之影響.............25
4.5.5. 飼料添加不同乳酸菌對白蝦吞噬指數之影響...........26
第五章、討論..........................................27
5.1. 飼料中乳酸菌菌量變化..............................27
5.2. 飼料中添加益生菌對白蝦成長之影響...................27
5.3. 飼料中添加乳酸菌對白蝦養殖池水水質改善之測試........27
5.3.1. 總氨氮.........................................27
5.3.2. 亞硝酸-氮......................................28
5.4. 飼料中添加益生菌對白蝦養殖池水菌量之影響...........28
5.4.1. 總菌數.........................................28
5.4.2. 弧菌數.........................................28
5.4.3. 乳酸菌數.......................................29
5.5. 飼料中添加乳酸菌對白蝦非特異性免疫之影響............29
5.5.1. 飼料添加不同乳酸菌對白蝦總血球數之影響............29
5.5.2. 飼料添加不同乳酸菌對白蝦酚氧化酵素 (PO)之影響.....29
5.5.3. 飼料添加不同乳酸菌對白蝦超氧陰離子 (O2-) 產生率之影響...................................................30
5.5.4. 飼料添加不同乳酸菌對白蝦吞噬率之影響.............30
5.5.5. 飼料添加不同乳酸菌對白蝦吞噬指數之影響...........31
第六章、結論..........................................32
參考文獻.............................................33
附錄.................................................64
附錄一、培養基成分....................................64
附錄二、緩衝溶液成分...................................67
附錄三：藥品配方......................................68

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