臺灣博碩士論文加值系統

澎湖縣是台灣目前箱網數量最多，產值最高的區域。產業發展卻有疾病感染之瓶頸，疾病會造成魚隻死亡及經濟上的損失。影響疾病的發生之要素有三:環境因素、病原菌因素、魚體體質因素。本實驗目的為調查箱網養殖區及非養殖區之環境，以菌相及其抗藥性等指標分析養殖對環境之影響，實驗方法則自94年6月至95年6月每月一次，調查澎湖縣兩家較具規模之箱網養殖場，每場三點箱網之海水採樣。採得之樣本則分別進行水質與微生物分析。水質分析的項目包含pH值、溶氧、水溫、鹽度、總油脂、總磷、氨氮、亞硝酸氮、硝酸氮等水質參數在時間上的改變。微生物分析則包括可培養菌數計算、生化鑑定以鑑定菌類及抗生素敏感性試驗分析。
調查結果顯示兩家養殖場的各項水質參數，數據相似。營養源方面，則以總磷的濃度除在其中幾月略超過甲類規範值外，平均值仍保持在甲類海域規範之內。兩場水中之分離出的病原性弧菌的百分比並無差異，其中箱網養殖海域分離株當中以Vibrio spp.為主要菌種（85%），其他以Vibrio alginolyticus（19%）、Vibrio harveiyi（9%）、Vibrio parahaemolyticus（8%）為主。
將分離到的Vibrio spp.以紙錠擴散法的藥物感受性試驗、結果發現弧菌菌樣對水產養殖常用之抗生素的抗藥性以amoxicillin（Resistant: 77%；
Mildly Sensitive : 4%）、ampicillin （R: 76%；M: 4%）、cephalothin （R: 56%；M: 15%）及sulfamethoxazole （R: 64%；M: 24%）之抗藥性比率最高（R: ＞ 50%）；gentamicin （R: 16%；M: 35%）、oxolinic acid （R: 10%；M: 29%）及nitrofurantoin （R: 8%；M: 19%）之抗藥性比率次之；而其他抗生素如 florfenicol（R: 0.4%；M: 5%）、chloramphenicol （R: 3%；M: 2%）、doxycycline （R: 2%；M: 3%）、oxytetracycline （R: 5%；M: 8%）、flumequine （R: 7%；M: 6%）之抗藥性比率較低。但二場之水環境甚至未養殖海域皆已都有抗藥性細菌之存在。
在此海域分離出的弧菌已對多種抗生素具有抗藥性，這可能顯示出澎湖地區有部分業者多年來使用抗生素的累積後果。在抗藥性菌株存在的情況下箱網養殖場抗生素將漸失效，導致抗生素使用量將更投高，而必導致疾病控制將必更困難。

Penghu County is the earliest area practicing cage culture in Taiwan, with its extensive and lengthy use of ocean site, consequently, disease outbreaks. Diseases not only cause death of fish, but also lead to substantial economical loses and non-sustainable production.

The occurance of fish diseases are affected by related three important factors: environment, health of fish, and pathogen. We tried to monitor the water quality, bacterium density and frequency of resistance to antibiotics in order to evaluate farming impact on the environment.
In this study, we survey two cage farms at different location for a period of one year to cover seasonal variation in Penghu area； collecting water samples for bacterial and water quality analysis. Our data suggested that the water quality parameters qualified the standard of first class aquaculture water condition. In term of bacterial content in the water of cage area; the bacteria total count and their species varied in seasons and areas. Among the major species of fish pathogen were vibrios that accounted for 85% of bacteria isolated. Among vibrios, Vibrio alginolyticus, V. harveyi and V. parahaemolyticus were the major population and accounted for 19%, 9% and 8% of vibrios, respectively. There is no clear correlation between the number of pathogen or species of pathogen with disease outbreak, except the bacteria number was higher in summer than that of the winter.

Results of antimicrobial susceptibility to twelve commonly used antibiotics indicated that the vibrios found in thees regions had a very high frequency of resistance to amoxicillin(Resistant: 77%；Mildly Sensitive : 4%), ampicillin(R: 76%；M: 4%), cephalothin(R: 56%；M: 15%), sulfamethoxazole(R: 64%；M: 24%); and medium resistance to gentamicin(R: 16%；M: 35%), oxolinic acid,(R: 10%；M: 29%), nitrofurantoin(R: 8%；M: 19%), and low frequency of resistance to florfenicol(R: 0.4%；M: 5%), chloramphenicol(R: 3%；M: 2%), doxycycline(R: 2%；M: 3%), oxytetracycline(R: 5%；M: 8%), flumequine(R: 7%；M: 6%).

We hope this study will be useful for the planning of farming management and the evaluation of the use of antibiotics in aquaculture.

中文摘要-------------------------------------------------------------------------------Ⅲ
英文摘要-------------------------------------------------------------------------------Ⅴ
目錄-------------------------------------------------------------------------------------Ⅶ
表目錄----------------------------------------------------------------------------------Ⅹ
圖目錄---------------------------------------------------------------------------------ⅩΙ
附錄----------------------------------------------------------------------------------ⅩⅢ
第一章 前言
第一節、研究背景………………………………………………………1
第二節、魚病問題………………………………………………………5
第三節、抗生素抗藥性…………………………………………………7
第四節、研究目的……………………………………………………12
第二章 研究方法與步驟
第一節、藥品與儀器…………………………………………………13
第二節、工作內容及實驗方法………………………………………14
第三節、海域水質監測方法…………………………………………18
第四節、海域微生物監測方法………………………………………22
第五節、藥物感受性試驗方法………………………………………26
第三章 結果
第一節、氣象統計……………………………………………………28
第二節、兩箱網養殖場之養殖紀錄…………………………………29
第三節、本研究在箱網養殖區進行水質環境監測…………………30
第四節、本研究在箱網養殖區進行海水環境微生物監測…………33
第五節、本研究在箱網養殖區進行海水中致病性弧菌密度分析…34
第六節、本研究在澎湖地區進行海水中弧菌對十二種藥物感受性百
分比…………………………………………………………39
第四章 討論
第一節、養殖環境與Stress部份………………………………………45
第二節、病原菌與流行疫情分析……………………………………47
第三節、箱網養殖水域微生物抗藥性現況…………………………50
第四節、養殖戶用藥習慣之探討……………………………………62
第五節、結語…………………………………………………………63
參考文獻……………………………………………………………………65

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