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姓名	許育瑄(Yu-hsuan Hsu)  查詢紙本館藏	畢業系所	環境工程研究所
論文名稱	藉由非抗性模式細菌對鎘之攝取機制探討量子點的生態毒性潛勢 (Probing the Potential Ecotoxicity of Quantum Dots through the Investigation of the Microbial Cadmium Uptake Mechanism)
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摘要(中)	奈米科技被譽為是二十一世紀最重要的科技之一，其蓬勃發展的結果雖可對各產業帶來劃時代的革命，但同時也免不期然的將會讓這些對生態造成何種影響仍不明確的新興物質排入環境中，使其發展的背後仍有隱憂。因此，了解奈米材料的傳輸、宿命及毒性，甚至更進一步地研究這類物質會對生態及人類健康帶來的衝擊為何，對於奈米科技的永續發展將極有助益，而探究人造奈米顆粒與微生物之間的相互作用則可視為推估這類材料潛在生態風險的第一步。然而，近期文獻雖已指出含鎘量子點在風化過程中，會因為鎘離子的溶出而對微生物產生制菌性或殺菌性的毒性效應，但文獻對於胞外化學物種組成在過程中如何影響量子點最後毒性作用的探討仍相當缺乏。有鑒於此，為釐清一般不具鎘抗性基因的原核細胞的鎘攝取機制，本研究利用革蘭氏陰性菌 Escherichia coli K-12 (ATCC 25404)為模式生物，以其存活率多寡做為鎘攝入的指標，在操縱試驗培養液化性的狀態下，探討胞外化學物種組成對於含鎘量子點所造成的微生物毒性之影響，並藉此推論及評估量子點潛在的生態衝擊程度。實驗結果證實原先設定的假說，即「溶解鎘主要以自由型態的鎘離子Cd2+進入菌株細胞中，且細胞對鎘的攝取過程主要依賴主動運輸此等機制」。這些結果意味著當水相系統維持在以自由態的鎘離子為優勢物種時，量子點將有可能對生態帶來最大的毒性衝擊。
摘要(英)	Recent advances in nanotechnology have created numerous and promising applications in all sectors of society; as a result, large scale developments of engineered nanomaterials (ENMs) have increased the likelihood of the release of these novel materials to the environment. Yet, the behavior of these nanomaterials in the environment is still poorly understood, thus raising the concerns of their potential ecological health risks. Given that microbes are the foundation of many ecosystems, a better understanding of the factors that control the microbial toxicity of ENMs is crucial for their sustainable use. To date, available data have indicated that toxicity of quantum dots (QDs) to bacteria is predominantly attributed to the release of toxic inorganic ions, in particular metal species, from weathered QD-cores. Hence, extracellular metal speciation is considered to play an important role in determining the ultimate toxicity of QDs towards microbial cells. In this study, we conducted exposure experiments to investigate the importance of cadmium (Cd) speciation in toxicity to pure cultures of Escherichia coli K-12 (ATCC 25404), a known Gram-negative strain lacking metal resistance czc genes in its genome and thus can be used as a model organism to represent generic non-resistant bacterial cells. Variable chloride chemistry experiments were carried out to modify Cd(II) speciation in assay media, and cell death or growth inhibition was used as an indicator of toxicity. Results show that the toxicity of Cd decreased along the chloride gradient where the severest growth inhibition occurred at the lowest salinity. Under the experimental conditions, inhibition was not a function of the total Cd(II) concentration but strongly correlated with free Cd2+ concentration. In addition, when strong chelating agents such as citrate and EDTA were supplemented to the assay medium, the Cd2+ concentrations were also modulated, resulting in reduced deleterious effects. As such, Cd toxicity to non-czc-mediated Cd resistant bacteria seemed to be explained by the free ion activity model (FIAM). We further examined if energy dependence of Cd uptake was required by strain K-12. Interestingly, differential viability was not observed when starved cells were exposed to media containing Cd(II) and variable chloride concentrations, suggesting that active transport may be the underlying uptake mechanism of Cd in this strain. Together, these results suggest that Cd toxicity to non-resistant bacterial cells is directly proportional to the uncomplexed, free Cd activity in solution, and uptake of Cd may be mediated via an energy-dependent transport system. Accordingly, on the basis of the predictions from the FIAM, these results also suggest that except for acidic and oligotrophic environments, under circumneutral and alkaline conditions cadmium-based QDs may not pose a significant threat to the ecosystem.
關鍵字(中)	★ 量子點 ★ 胞外化學物種組成 ★ 原核微生物 ★ 鎘攝取機制 ★ 微生物毒性	關鍵字(英)	★ quantum dots (QDs) ★ extracellular chemical speciation ★ prokaryote ★ mechanism of Cd(II) uptake ★ microbial toxicity
論文目次	摘要 I ABSTRACT II 目錄 IV 圖目錄 VII 表目錄 XI 第一章 前言 1 1.1 研究動機 1 1.2 研究目的 5 第二章 文獻回顧 6 2.1 奈米科技 6 2.1.1奈米顆粒對環境可能的影響 7 2.1.2 量子點基本介紹與含鎘量子點 8 2.1.3 量子點的毒性機制 11 2.2微生物在人造奈米材料毒理研究的重要性 14 2.3 鎘的化學、生物化學與毒性介紹 17 2.3.1 鎘的基本特性與應用 18 2.3.2環境中的鎘與化學物種組成 19 2.3.3 鎘的細胞毒性 21 2.4化學物種組成與生物有效性 24 2.4.1自由離子活性模型(free ion activity model , FIAM) 24 第三章 實驗方法 27 3.1 實驗藥品與儀器 27 3.1.1實驗藥品與試劑 27 3.1.2實驗儀器 28 3.2 化學物種組成模擬軟體 29 3.3 實驗用模式生物 30 3.4 試驗培養液的成分及製備 30 3.5 E. COLI懸浮菌液準備 38 3.6 微生物毒性試驗方法 38 3.6.1 平板計數法 41 3.6.2 光學密度生長曲線法 41 3.6.3 live/dead微生物存活率試驗 42 3.7 測量胞內外鎘含量之細胞洗滌試驗 44 第四章 結果與討論 47 4.1 ASSAY MEDIUM之物種組成分佈曲線 47 4.2試驗培養液之最低抑菌濃度 54 4.3 鎘物種組成對E. COLI的毒性影響探討 57 4.3.1 不同氯離子濃度下毒性試驗結果探討 57 4.3.2 不同citrate濃度下毒性試驗結果探討 63 4.4 不同PH下毒性試驗結果探討 66 4.4.1 小結 76 4.5 E. COLI對鎘的攝取方式 77 4.6測量進入細胞中的鎘之探討 80 4.7菌存活率試驗 85 4.8環境意義 91 第五章 結論與建議 96 5.1 結論 96 5.2 建議 97 參考文獻 98
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指導教授	林居慶(Chu-ching Lin)	審核日期	2015-8-26
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