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研究生:周廷耀
研究生(外文):Tin-Yao Chou
論文名稱:碳源的添加對等鞭金藻增殖的影響
論文名稱(外文):The influence of carbon addition on the growth of microalga Isochrysis galbana
指導教授:蘇惠美蘇惠美引用關係
學位類別:碩士
校院名稱:國立中山大學
系所名稱:海洋生物研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:46
中文關鍵詞:等鞭金藻pH迴饋控制微藻碳酸氫鈉二氧化碳
外文關鍵詞:Isochrysis galbanacarbon dioxidecarbonpH feedbacksodium bicarbonate
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對大多數藻類而言,在天然的水域中,二氧化碳的濃度是充足的,但在人工培養的情況下,藻類的密度很高,二氧化碳的供應往往不足,成為光合作用的限制因子,進而影響藻細胞的生長繁殖。本研究嘗試小量靜置培養用添加二氧化碳的方式,調整初始pH至7.5、6.5和5.5(CO2-7.5i、CO2-6.5i、CO2-5.5i),且以鹽酸調整至相同pH(HCl-7.5i、HCl-6.5i、HCl-5.5i)及都未添加 (blank) 為對照,連續照光養殖。結果為第二至第五天的細胞數都以添加二氧化碳至初始pH為6.5的最佳,5.5的最差。再選用CO2-6.5i、HCl-6.5i和blank三組,用每日調回pH的方式,亦有相同的結果,以CO2-6.5e的最佳,第三天的細胞數為blank的221 %。另一個小量靜置培養為添加碳酸氫鈉,起始濃度為1 g/l、0.5 g/l、0.1 g/l、0.05 g/l、0.01 g/l和未添加的blank,在五天的養殖期間,每天的細胞數除1 g/l較低外,其餘五組無顯著差異。再選用 0.5 g/l為起始碳酸氫鈉濃度,調整初始pH為6.5(SBH),以不調整pH (SB)及都未處理(Blank)為對照,SBH第二至第五天有最大的細胞數,為blank的212 %。大量打氣式培養則採用二氧化碳迴饋控制的方式,控制藻水的pH為6.5、7.0和7.5,另以鹽酸迴饋控制pH為7.5和未控制為對照,重覆實驗兩次,兩次結果為添加二氧化碳的三組間,細胞的增殖雖無差異,但都明顯高於鹽酸控制組和未控制組,最大細胞數於第七天左右可達1100-1400 x 104 個/ml,約為未控制組的兩倍,且藻細胞數達到最大值時,水中的氮源剛好用盡,而磷源還有餘。本研究顯示等鞭金藻在良好的培養環境下,適量碳源的添加及維持適當的pH,可以使此藻有較快的生長和獲得較高的藻細胞濃度。
The concentration of carbon dioxide in natural seawater is sufficient for microalgal growth, but insufficient for high algal-density culture due to limitation of photosynthesis in the artificial medium. This study was adjusted the initial pH to 5.5, 6.5 and 7.5 by adding CO2 or HCl in 1L flask cultured stagnantly with continuous illumination. The best growth of Isochrysis galbana was found in the culture (CO2-6.5i) with initial pH 6.5 using CO2 adjustment and maintaining the same during day 2 to day 5, while the worst was in CO2-5.5i. Furthermost, we adjusted the pH of cultures daily to the set values; the best growth was also found in CO2-6.5e having the cell number 221% of the blank. Initial addition of NaHCO3 with doses of 0.01 g/L, 0.05 g/L, 0.1 g/L, 0.5 g/ L and 1g/ L in the culture, showed the lowest cell number after 5 days culture is in the group of 1.0 g/L and no significant difference among the rest groups. Addition of 0.5 g/L NaHCO3 and adjusted the pH to 6.5 by HCl in the beginning promoted algal growth and resulted in the culture having 212 % cell number of the blank. Using the feedback control system, 100 L algal cultures with aeration and providing CO2 gas or HCl liquid to maintain the pH as 5.5, 6.5 and 7.5 or 7.5 individually were conducted to test the effect of pH control on the mass culture of I. galbana. Better growth was found in the culture with CO2 feedback control than HCl-control in duplicate experiments. It also showed significant difference among the groups adjusted pH between 6.5-7.5. The cell concentration could reach 1100-1400 x 104 cells/ml and was about double the amount of the blank without pH control cultured in 7 days. Meanwhile, the NO3-N concentration was nearly exhausted while the PO4-P still replete. This study reveals the high concentration and fast growth of I. galbana can be maintained under the suitable physical condition providing the carbon source in an optimal pH.
中文摘要………………………………………………………………..Ⅰ
英文摘要………………………………………………………………..Ⅱ
總目錄…………………………………………………………………..Ⅲ
表目錄…………………………………………………………..………Ⅵ
圖目錄…………………………………………………………………..Ⅸ
附錄目錄……………………………………………………………….Ⅹ
壹、前言……………………………………………………………….…1
一、微藻的功用……………………………………………………….…1
(一)、微藻在水產養殖的應用…………………………………………..1
(二)、微藻在醫藥食品的應用………………………………………..…3
(三)、微藻在污染上的應用……………………………………………...4
二、微藻成長與營養鹽的關係……………………………………….….6
(一)、微藻成長與碳的關係……………………………………………...6
(二)、微藻成長與氮的關係……….……………………………………..8
(三)、微藻成長與磷的關係……………………………………………..9
(四)、微藻成長與pH的關係……………………...……………………10
三、等鞭金藻的相關研究……………………………………………....11
四、本研究探討的方向…………………………………………………12
貳、材料方法……………………………………………………………14
一、實驗材料與實驗生物的來源與培養………………………………14
(一)、等鞭金藻大溪地株(Isochrysis aff. galbana )及培養設備………14
(二)、韋音培養液……………………………………………………….14
(三)、培養用海水處理………………………………………………….15
(四)、CO2氣體的裝置…………………….…………………………….15
(五)、pH迴饋控制…………………………………………………. 16
二、實驗方法……………………………………………………….…..15
(一)、小量靜置培養…………………………………………………….16
1. 起始添加二氧化碳和鹽酸試驗…………………………………….16
2. 每日添加二氧化碳和鹽酸試驗………………………………….…17
3. 碳酸氫鈉試驗…………………………………………………...…..17
4. 碳酸氫鈉加酸試驗……………………………………………...…..17
(二)、大量培養……………………………………………………..….18
1. 打氣量試驗……………………………………………………….…19
2. 接種濃度試驗………………………………………………….……19
3. pH迴饋控制試驗…………………………...…………………..…...19
(三)、營養鹽測定…………………………………………………..…..20
1. 海水中亞硝酸鹽(N02-)含量測定…………………………...…....…20
2. 海水中硝酸鹽(NO3-)含量測定………………...………………...…21
3. 海水中銨(NH4+)含量測定……………………………………….….21
4. 海水中無機磷(PO43-)含量測定………………………………..…....22
5. 總鹼度測定……………………………………………………….…22
(四)、增殖率和細胞密相對百分比的計算……………………………23
1. 增殖率………………………………………………………………..23
2. 細胞密度相對百分比………………………………………………..24
(五)、統計分析…………………………………………………………24
參、結果……………………………………………………………...….25
一、1l小量靜置培養………………………………………………..….25
(一)、起始添加二氧化碳和鹽酸試驗……………………………….…25
(二)、每日添加二氧化碳和鹽酸試驗………………………………….26
(三)、碳酸氫鈉試驗……………………………………………….……27
(四)、碳酸氫鈉加酸試驗………………………………………….……28
二、大量培養試驗………………………………………………………29
(一)、不同打氣量試驗………………………………………………….29
(二)、不同接種濃度試驗……………………………………………….30
(三)、pH迴饋試驗………………………………………………….…..31
肆、討論………………………………………………………………....34
一、起始添加和每日添加二氧化碳和鹽酸試驗………………...…….35
二、碳酸氫鈉和碳酸氫鈉加酸試驗……………………………………37
三、pH迴饋試驗………………………………………………….…….39
伍、參考文獻…………………………………………………………....46
陸、附錄……………………………
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