抗生素耐藥性定義為微生物在抗生素的影響下生存的能力，全球範圍內抗生素被濫用的情形，即使在衛生組織不斷警告下，仍未改善。而這一情況，導致抗生素耐藥性在微生物間傳播，也逐漸的演變成多重耐藥性，而現代醫學則加快了這轉變的過程，優勝劣敗，淘汰掉對於藥物較敏感的微生物，而自然的留下了耐藥菌繼續進行繁衍及傳播。而抗生素的殘留物抑或是衍生物會經過許多渠道流入自然環境中，其中廢水處理廠更是隱憂之一，如果廢水處理的不完善，更可能會導致廢水中擁有多重耐藥性的微生物去汙染自然界中的微生物，又因為自然環境符合許多抗生素耐藥性傳播的必要條件，因此更會加快微生物間傳播耐藥性現象的加劇。

Antibiotic resistance is defined as the ability of microorganisms to survive under the influence of antibiotics. The situation of antibiotic abuse worldwide has not improved even with constant warnings from WHO. This situation has led to the spread of antibiotic resistance among microorganisms, and gradually evolved into multi-drug resistance, and modern medicine has accelerated the process of this transformation, eliminating the microorganisms that are sensitive to drugs. Naturally, drug-resistant bacteria are left to continue multiplying and spreading. Antibiotic residues or derivatives will flow into the natural environment through many channels, and plants of wastewater treatment are one of the worries. If the wastewater treatment is imperfect, it may pollute thenature by microorganisms with multiple drug resistance. The natural environment meets many necessary conditions for the spread of antibiotic resistance, so it will accelerate the spread of resistance among microorganisms.

摘要 i
ABSTRACT ii
誌謝 iii
目錄 iv
表目錄 vii
圖目錄 viii
第1章 緒論 1
1.1 前言 1
1.2 研究背景及動機 2
第2章 文獻回顧 3
2.1 抗生素的介紹 3
2.2抗生素的機制 5
2.3抗生素耐藥性 6
2.4水平基因轉移( Horizontal Gene Transfer , HGT ) 7
2.4.1接合作用 8
2.5抗生素汙染與汙水處理廠的關係 8
2.6抗生素在環境中的影響 9
2.7 Metagenomics Next generation Sequencing ( mNGS ) 10
2.8菌種鑑定 10
2.9綠色螢光蛋白(Green Fluorescent Protein, GFP) 11
2.10抑菌圈 11
2.11 pCF10 11
第3章 研究方法 12
3.1 實驗儀器與材料 12
3.1.1 實驗儀器 12
3.1.2 藥品成分 13
3.1.3 培養基配方 14
3.1.4 藥品配方 14
3.2 實驗材料 15
3.3 實驗方法 16
3.3.1大腸桿菌轉型方法(E.coli Transformation) 16
3.3.2抽取廢水中細菌DNA 18
3.3.3 Metagenomics Next Generation Sequencing 19
3.3.4 菌種鑑定 19
3.4 實驗流程 20
3.4.1 廢水處理 20
3.4.2圓盤擴散法( 抑菌圈 ) 20
3.4.3 廢水菌之接合作用 21
3.4.4 處理後之廢水菌與MG1655 pHL 1278接合作用 21
3.4.5 菌種鑑定後之大腸桿菌與HL4195 pHL1278接合作用 22
3.4.6 多種抗藥性 23
第4章 結果與討論 27
4.1廢水菌之間的接合作用實驗 27
4.2 處理後之廢水菌與MG1655 pHL 1278接合作用實驗 28
4.3菌種鑑定 29
4.3.1 菌種鑑定後之大腸桿菌與HL4195 pHL1278接合作用 29
4.4尋找廢水菌內抗藥性基因轉移無法再現原因 30
4.4.1 圓盤擴散法 30
4.4.2 多種抗藥性 31
4.5 Metagenomics Next Generation Sequencing 33
第五章 結論 37
參考文獻 38

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