臺灣博碩士論文加值系統

本研究延續Chen and Lin 於1997年所提出的連續式試驗方法進行藻類(Selenastrum capricornutum)重金屬毒性試驗。經由稀釋方法，將每次實驗時所需的藻液使用量由原系統的100 % 降為50 % 及6 % 後進行毒性試驗，以探討藻類細胞密度與試驗敏感度間之相對應關係。而由實驗結果發現: 當降低藻類細胞密度時，實驗所得之 EC50值、LOEC值及NOEC值皆隨之下降，亦即試驗敏感度隨之提高。而由於藻液使用量的降低，連續式試驗之實驗週期可由原系統的4天進一步縮短為2天甚至1天，可有效增加試驗頻率。
由於目前各藻類標準方法有逐漸禁止於試驗基質中添加螫合物的趨勢，因此在本研究中亦進行不含(或減量)螫合物-EDTA的相關實驗，藉以探討EDTA含量對連續式培養下之藻類生長及對連續式藻類毒性試驗結果所造成的影響。同時為對照之用，研究中亦操作不含EDTA的批次式藻類毒性實驗。根據結果顯示: 在將進流基質中的EDTA含量由30 μg/L降為15 μg/L後，連續式培養下之藻類在長時間下仍可維持正常之生長。而不含EDTA之批次式培養下，藻類細胞生長則明顯受到抑制。至於進行批次式及連續式藻類毒性試驗時，EDTA的減少將造成兩者在毒性試驗敏感度大幅提高。
最終，對各不同條件下之藻類毒性試驗進行參考毒物Cd2+之再現性比較。在50 % 細胞密度之連續式試驗中，實驗之再現性明顯優於批次式試驗。然而在6 % 之試驗中，由於由於藻類細胞數目較少，些微的細胞數量變動即可導致生長率產生大幅度變化，因此實驗之再現性較為不佳。

In this study , we performed the modified method of the continuous algal(Selenastrum capricornutum) toxicity test which was introduced by Chen & Lin in 1996 . By diluting , we decreased the use of algal cells per ml from 100 % to 50 %、6 % in each continuous experiment . We then researched the correlation between the algal cell density with the sensitivity of the continuous toxicity test . And we found that : When we lowered the algal cell density , the sensibility would be improved markedly i.e. the EC50 values 、NOEC values and LOEC values dropped off . In addition , the lower cell density could shorten the experimental period from 4 days to two or one day . The short period could increase the frequency of the operation of the experiment .
Because the new algal standard batch method is intend to inhibit the use of the chelator-EDTA in the medium , we also proceeded the correlative experiments which not including (or reduced) EDTA . Then , we researched how the lack of EDTA could effect on the algal growth , and compared the results of the toxicity test from the exist of EDTA or not . Additionally , we proceeded the batch toxicity test without EDTA in comparison to the results from continuous toxicity test. Then we found that : when the EDTA , in the medium , was reduced from 30 μg/L to 15 μg/L , the culture of algae in the continuous incubator was still maintained in healthy condition for a long time . In the toxicity test , however, when we reduced the use of the EDTA , the sensibility would be increased . Finally , we compared the reproducibility in each different toxicity test .

目錄
表次頁次
中文摘要Ⅰ
英文摘要Ⅲ
誌謝Ⅴ
目錄Ⅵ
表目錄Ⅷ
圖目錄Ⅸ
第一章 緒論1
1.1 研究緣起1
1.2 研究目的4
第二章 文獻回顧5
2.1 試驗藻種5
2.2 藻類的培養方式7
2.2.1 批次式培養技術7
2.2.2 連續式培養技術8
2.3 pH的控制10
2.4 植種數量11
2.5 毒性物質12
2.6 實驗培養基13
2.6.1 氮、磷的影響13
2.6.2 螫合物的影響15
第三章 基本理論17
3.1 濃度-反應曲線17
3.2 基本生長動力學18
3.3 數據分析19
第四章 實驗設備與方法23
4.1 實驗設備及藥品23
4.2 試驗藻種28
4.3 培養基28
4.4 試驗毒物29
4.5 實驗條件的控制31
4.6 實驗前的準備31
4.7 實驗步驟34
4.8 生長率(μ)之計算35
4.9 數據分析35
第五章 結果與討論39
5.1 連續式系統培養槽藻液取用量與實驗週期之關係39
5.2 於固定藻類初始胞密度(50 %)下，改變毒性試驗系統進流基質
之氮、磷濃度對生長率之影響42
5.3藻類之重金屬毒性試驗45
5.3.1 批次式及連續式實驗之EC50值比較45
5.3.2 不同初始細胞密度下，重金屬Cd2+對連續式實驗之藻類
生長影響48
5.4 EDTA添加量對藻類重金屬毒性試驗之影響51
5.4.1 EDTA含量對連續式(藻類培養系統)培養下之藻類生長的
影響51
5.4.2 EDTA含量對批次及連續式(毒性試驗系統)培養下之藻類
生長的影響57
5.4.3 降低EDTA含量對藻類重屬毒性試驗結果之影響59
5.5 EE50值與EC50值之比較65
5.6 各實驗之再現性研究69
第六章 結論與建議71
參考文獻75
附錄一 Ecological Effects Test Guidelines 摘要
(Algal Toxicity)80
附錄二 連續式藻類之重金屬毒性試驗(6 %細胞密度)82
附錄三 連續式藻類試驗數據87
附錄四 批次式藻類試驗數據(No-EDTA)99
表目錄
表次頁次
表2.1 批次式與連續式藻類毒性試驗系統優缺點比較9
表2.2 各藻類培養基組成份分析比較14
表4.1 以離子分析儀及分光光度計之偵測極限27
表4.2 藻類培養基之巨量營養組成份30
表4.3 藻類培養基之微量營養組成份30
表4.4 Coulter Counter電子計數器設定條件33
表5.1 進流基質中之氮、磷含量對細胞生長的影響44
表5.2 比較於不同藻類毒性試驗方法下，所得EC50值之差異性46
表5.3 連續式藻類毒性試驗(Cd2+)之實驗相關數據(50 %初始細胞密度)49
表5.4 連續式藻類毒性試驗(Cd2+)之實驗相關數據(100 %初始細胞密度)49
表5.5 標準批次式(U.S.EPA)及本研究修正過之藻類培養基組成分比較52
表5.6 比較在不同EDTA進流量下，連續式培養達平衡時之差異情形56
表5.7 EDTA含量對批次式及連續式(毒性試驗系統)培養之下藻類生長
的影響58
表5.8 比較EDTA含量變化對批次式及連續式藻類重金屬毒性試驗之
影響61
表5.9 連續式藻類毒性試驗(Cd2+)之實驗相關數據(減量-EDTA)63
表5.10 比較連續式藻類毒性試驗Cd2+及Ni2+之NOEC及LOEC值64
表5.11 比較不同初始細胞密度下之連續式藻類Cd2+及Ni2+毒性試驗
EE50值66
表5.12 以EE50值比較EDTA含量變化對批次式及連續式藻類重金屬
毒性試驗之影響68
表5.13 比較不同藻類試驗方法下，Cd2+之試驗再現性70
表5.14 比較藻類在不同初始細胞密度下EC50值之差異性(30μg/L-EDTA)84
表5.15 比較藻類在不同初始細胞密度下EC50值之差異性(減量-EDTA)85
表5.16 比較連續式藻類毒性試驗(6 %初始細胞密度)在不同EDTA含量下，重金屬Cd2+之NOEC及LOEC值86
圖目錄
圖次頁次
圖2.1 Selenastrum capricornutum之藻類圖鑑6
圖4.1 連續式藻類培養系統流程圖24
圖4.2 連續式毒性試驗系統概圖24
圖4.3 毒性試驗數據分析流程圖37
圖4.4 Visual Basic之Project程式選取所需資料並加以分析38
圖5.1 培養槽之藻液取出量與穩定達成時間兩者之關係41
圖5.2 在50 %初始細胞密度下，毒性試驗系統之氮、磷含量改變對藻類
細胞生長的影響(24hr)44
圖5.3 連續式培養系統(15 μg/L-EDTA)藻類細胞密度隨時間變化情形54
圖5.4連續式培養系統(15 μg/L-EDTA)藻類平均細胞體積隨時間之變化54
圖5.5 連續式培養系統(15 μg/L-EDTA)藻類細胞總體積隨時間之變化55
圖5.6 連續式培養系統(15 μg/L-EDTA)pH值隨時間之變化情形55

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