轉機因生物的研究.doc
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轉機因生物的研究,頁數(shù)：43字數(shù)：26739摘要1abstract21.前言31.1 轉基因生物安全性評價的必要性及評價體系的建立現(xiàn)狀31.2 評價轉bt基因抗蟲棉對魚類安全性的意義41.3 對魚類安全性的評價方法71.4 有關斑馬魚的研究102 材料和方法142.1 材料142.1.1 實驗動物142.1.2 實驗用棉...
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轉機因生物的研究
頁數(shù)：43 字數(shù)：26739
摘要 1
ABSTRACT 2
1. 前言 3
1.1 轉基因生物安全性評價的必要性及評價體系的建立現(xiàn)狀 3
1.2 評價轉Bt基因抗蟲棉對魚類安全性的意義 4
1.3 對魚類安全性的評價方法 7
1.4 有關斑馬魚的研究 10
2 材料和方法 14
2.1 材料 14
2.1.1 實驗動物 14
2.1.2 實驗用棉籽 14
2.1.3 試劑 14
2.1.4儀器 15
2.2 方法 17
2.2.1 實驗處理 17
2.2.2急性毒性實驗 18
2.2.3 慢性毒性實驗 18
3 結果 20
3.1 急性毒性實驗 20
3.2 慢性毒性實驗 20
3.2.1器官和組織的形態(tài)、結構
20
3.3 微核試驗 24
3.3.1陽性對照物試驗 24
3.3.2 正式實驗 24
3.3.3 紅細胞的測量 27
4. 討論 28
4.1急性毒性試驗 28
4.2慢性水平的毒性試驗 29
4.3微核試驗 31
4.4結論： 33
參考文獻 34
附錄Ⅰ 39
附錄Ⅱ 40
附錄Ⅲ 41
圖版說明 42
摘要
以斑馬魚(Brachydanio rerio)為實驗材料，從一般毒性（急性，慢性），遺傳毒性兩個層次評價轉Bt基因抗蟲棉棉籽粉對魚類的生物安全性。急性毒性試驗結果顯示轉Bt基因抗蟲棉對斑馬魚無急性致死毒性，也沒有急性非致死性毒性。慢性毒性試驗的病理學檢查表明，肝臟、脾臟和腸均未發(fā)現(xiàn)解剖觀察水平和光鏡水平的病變。采用斑馬魚外周血紅細胞微核試驗的遺傳毒性方法，證明誘變劑氟化鈉可以明顯誘導斑馬魚外周血紅細胞微核率升高，并確定了合適的陽性反應條件。運用改進的小型魚采血方法和建立的斑馬魚微核試驗檢測方法，檢測了轉Bt基因抗蟲棉的細胞遺傳毒性，結果表明轉Bt基因抗蟲棉沒有誘發(fā)斑馬魚紅細胞微核的遺傳毒性。實驗結果表明在魚類飼料中使用規(guī)范比例的轉Bt基因棉籽粉是安全的。
關鍵詞：轉Bt基因抗蟲棉,斑馬魚,安全性評價
Abstract
The biosafety of transgenic Bt cottonseed to fish is assessed by means of regular toxicity test (acute test and chronic test) and genetoxicity test. Zebrafish (Brachydanio rerio) is chosen as model experimental animal. No acute lethal toxicity and other acute toxicity of transgenic Bt cottonseed to zebrafish is observed. After chronic toxicity test, liver, spleen and intestine of all experimental groups are examined with help of pathological methods. No pathological change on morphology level and on microscopic level is observed. Micronucleus test (MNT) is chosen on behalf of genetoxicity test. NaF is found out to be a kind of mutagens to zebrafishes and appropriate conditions for NaF to induce Micronucleus (MN) in peripheral erythrocytes is also set up. The size of peripheral erythrocytes, their nuclei, and MN is measured. The size of them shows that peripheral erythrocytes of zebrafishes are a kind of good materials for MNT. The result of MNT shows that transgenic Bt cottonseed does not induce raise of MN ratio significantly.
On base of the experimental results, a primary conclusion is made that the regular proportion of transgenic Bt cottonseed in fish meal is safe to fishes.
Key words: Transgenic Bt cottonseed, Zebrafish, Biosafety assessment
頁數(shù)：43 字數(shù)：26739
摘要 1
ABSTRACT 2
1. 前言 3
1.1 轉基因生物安全性評價的必要性及評價體系的建立現(xiàn)狀 3
1.2 評價轉Bt基因抗蟲棉對魚類安全性的意義 4
1.3 對魚類安全性的評價方法 7
1.4 有關斑馬魚的研究 10
2 材料和方法 14
2.1 材料 14
2.1.1 實驗動物 14
2.1.2 實驗用棉籽 14
2.1.3 試劑 14
2.1.4儀器 15
2.2 方法 17
2.2.1 實驗處理 17
2.2.2急性毒性實驗 18
2.2.3 慢性毒性實驗 18
3 結果 20
3.1 急性毒性實驗 20
3.2 慢性毒性實驗 20
3.2.1器官和組織的形態(tài)、結構
20
3.3 微核試驗 24
3.3.1陽性對照物試驗 24
3.3.2 正式實驗 24
3.3.3 紅細胞的測量 27
4. 討論 28
4.1急性毒性試驗 28
4.2慢性水平的毒性試驗 29
4.3微核試驗 31
4.4結論： 33
參考文獻 34
附錄Ⅰ 39
附錄Ⅱ 40
附錄Ⅲ 41
圖版說明 42
摘要
以斑馬魚(Brachydanio rerio)為實驗材料，從一般毒性（急性，慢性），遺傳毒性兩個層次評價轉Bt基因抗蟲棉棉籽粉對魚類的生物安全性。急性毒性試驗結果顯示轉Bt基因抗蟲棉對斑馬魚無急性致死毒性，也沒有急性非致死性毒性。慢性毒性試驗的病理學檢查表明，肝臟、脾臟和腸均未發(fā)現(xiàn)解剖觀察水平和光鏡水平的病變。采用斑馬魚外周血紅細胞微核試驗的遺傳毒性方法，證明誘變劑氟化鈉可以明顯誘導斑馬魚外周血紅細胞微核率升高，并確定了合適的陽性反應條件。運用改進的小型魚采血方法和建立的斑馬魚微核試驗檢測方法，檢測了轉Bt基因抗蟲棉的細胞遺傳毒性，結果表明轉Bt基因抗蟲棉沒有誘發(fā)斑馬魚紅細胞微核的遺傳毒性。實驗結果表明在魚類飼料中使用規(guī)范比例的轉Bt基因棉籽粉是安全的。
關鍵詞：轉Bt基因抗蟲棉,斑馬魚,安全性評價
Abstract
The biosafety of transgenic Bt cottonseed to fish is assessed by means of regular toxicity test (acute test and chronic test) and genetoxicity test. Zebrafish (Brachydanio rerio) is chosen as model experimental animal. No acute lethal toxicity and other acute toxicity of transgenic Bt cottonseed to zebrafish is observed. After chronic toxicity test, liver, spleen and intestine of all experimental groups are examined with help of pathological methods. No pathological change on morphology level and on microscopic level is observed. Micronucleus test (MNT) is chosen on behalf of genetoxicity test. NaF is found out to be a kind of mutagens to zebrafishes and appropriate conditions for NaF to induce Micronucleus (MN) in peripheral erythrocytes is also set up. The size of peripheral erythrocytes, their nuclei, and MN is measured. The size of them shows that peripheral erythrocytes of zebrafishes are a kind of good materials for MNT. The result of MNT shows that transgenic Bt cottonseed does not induce raise of MN ratio significantly.
On base of the experimental results, a primary conclusion is made that the regular proportion of transgenic Bt cottonseed in fish meal is safe to fishes.
Key words: Transgenic Bt cottonseed, Zebrafish, Biosafety assessment
