臺灣博碩士論文加值系統

中文摘要
本研究分別選定基隆和平島八尺門岸邊及基隆嶼外島之岸邊為實驗地點，以檜木片、FRP片、鋼片及玻璃片為材料，在海面下一公尺進行懸吊片試驗，探討真菌附著狀況。八尺門海水與基隆嶼海水之水溫、pH值與溶氧量之差異不大，水溫為17~25oC，pH值為8.1~8.5，溶氧量為7.5~8.2 ppm，但真菌含量則以八尺門含量較高。八尺門海水和基隆嶼海水總有機碳 (TOC) 和氨氮含量，二者分別為494和337 ppm (TOC)，以及0.57和0.37 ppm (氨氮)。分別針對不同地點的真菌含量作比較，以八尺門的含量高於基隆嶼；而季節變化的影響以冬季附著量較高；材料片種類對真菌附著之影響隨季節而不同。至於四種不同材料片上附著真菌種類，並無明顯不同。附著酵母菌分別屬於下列八屬：Candida、Rhodotorula、Cryptococcus、Trichosporon、Pichia、Klosckera、Saccharomyces及Debaryomyces。已鑑定之附著黴菌主要包括Penicillium、Geotrichum、Mucor、Aspergillus、Fusarium、Trichoderma與Cladosporium。由懸吊片所得269株真菌分離株，挑選出生長及附著能力最佳之三株菌Candida guilliermondii 379、Candida humicola 423及Debaryomyces polymorphus 471。另外，Rhodotorula rubra 091雖然附著力不高，但其在懸吊片上出現頻率很高，本研究乃以這四株海洋酵母菌為測試菌株，探討其在海水中生存，以及環境因子如碳源、氮源、無機鹽、pH值及溫度對其生長及對玻璃片附著能力之影響。此四株酵母菌在自然海水中可增殖100~1000倍，且在海水中存活110天以上。海水pH值 (6-9) 及溫度 (20-37oC) 對其生長及附著能力影響不顯著。海水中碳源、氮源及鹽類對酵母菌附著力之影響會隨菌株不同而異。海水中添加0.5% 碳源可能降低菌株附著力，但添加0.1% 纖維素、酪蛋白與硝酸鉀有助於菌株附著。Ca2+和Mg2+亦有助於菌株附著。另由酵母菌C. guilliermondii 379與海水附著細菌分離株Psedumonas cepacia在海水中所進行之試驗，顯示酵母菌與細菌同時存在，對彼此之間的附著影響不大；但固體表面若先有細菌附著，會降低酵母菌附著力；反之酵母菌之附著並不影響細菌的附著力。以MATH分析酵母菌體表面疏水性得知在Brain Heart Infusion Broth (BHIB) 中生長之Candida guilliermondii與Debaryomyces polymorphus，菌體表面疏水性隨菌齡增加而增大，其附著力亦隨之增大。但Candida humicola及Rhodotrula rubra卻得相反結果，表面疏水性及附著力皆隨菌齡增加而下降。海水中C. guilliermondii之表面疏水性隨時間增加而遞減，成為疏水性很低之現象。然其附著力卻隨時間增加而增大，顯示菌體表面電荷，可能為影響海水中酵母菌附著力的主要決定因子，而非疏水性。

Abstract
Substrata of wood, fiber-reinforced plastic (FRP), steel and glass were immersed in seawater under 1 meter of sea level at the seashores of Pa-Zhi-Men and Keelung Isle. The amounts and flora of the attached fungi on these substrata were investigated. The temperature, pH value and dissolved oxygen of seawater from these two different experimental areas were similar, which were in the ranges of 17~25oC, 8.1~8.5 and 7.5~8.2 ppm, respectively. However, higher fungal content was found in seawater at Pa-Zhi-Men. The total organic contents in seawater at Pa-Zhi-Men and Keelung Isle were 494 and 337 ppm, respectively; while ammonium nitrogen contents being 0.57 and 0.37, respectively. The effects of location and season on the fungal attachment were found. Higher counts of attached fungi were obtained at Pa-Zhi-Men than at Keelung Isle. Higher fungal counts were observed in winter than in summer. The effect of the types of substrata on the fungal attachment was various with season. The fungal flora on the tested substrata was similar. The attached yeast isolates were included in the genera of Candida, Rhodotorula, Cryptococcus, Trichosporon, Pichia, Klosckera, Saccharomyces and Debaryomyces. The identified mold isolates were mainly in the genera of Penicillium、Geotrichum、Mucor、Aspergillus、Fusarium、Trichoderma and Cladosporium. Three fungal isolates, Candida humicola 423, Candida guilliermondii 379 and Debaryo polymorphus 471, were chosen as the tested microorganisms due to their high adhesive capability among 269 fungal isolates. In addition, one strain of Rhodotorula rubra 091 was also tested due to its high incidence on the substrata. Their proliferation in seawater and the environmental factors of seawater, such as carbon and nitrogen, salt contents, pH value and temperature, on the growth and attachment on glass slide were investigated. These 4 isolates could proliferate by 100-1000 times and survived over 110 days in seawater. Temperature in the ranges of 20-37oC and pH value (6-9) did not affect their growth and adhesive capability. The effects of carbon sources and nitrogen sources in seawater varied with the tested isolates. In general, the addition of 0.5% various carbon sources obviously decrease the fungal attachment. 0.1% cellulose, casein or KNO3 in seawater stimulated yeast adhesion. Cations of Ca2＋ and Mg2＋ also showed positive effects for fungal attachment. The co-existence of yeast C. guilliermondii and bacterium Pseudomonas cepacia did not affect their individual adhesive ability. However, the pre-existence of bacteria on glass slide decreased the adhesive ability of yeast. The bacterial attachment was similar, irrespective of the presence of yeast. When C. guilliermondii and D. polymorphus were cultivated in brain heart infusion broth (BHIB), the cell surface hydrophobicity, measured by microbial adhesion to hydrocarbon method (MATH), and adhesive ability increased with increasing of cell age. However, the surface hydrophobicity and adhesive ability of C. guilliermondii and R. rubra decreased as their cell ages increased. On the other hand, the cell surface hydrophobicity of C. guilliermondi in seawater gradually decreased and became very small; however, its adhesive ability increased. This demonstrated that the factor of the cell surface charge may become the determinant factor for the attachment of yeast in seawater.

目 錄
中文摘要- ----------------------------------------------------------------------------I
英文摘要-- --------------------------------------------------------------------------II
前言 ---------------------------------------------------------------------------------IV
第一章 文獻回顧 ---------------------------------------------------------------1
一、生物膜的定義 ---------------------------------------------------------------1
二、生物膜之形成 ----------------------------------------------------------------1
三、影響微生物附著的因子 ---------------------------------------------------2
四、真菌在微生物附著中的角色 ---------------------------------------------6
五、影響真菌附著之因子 ------------------------------------------------------7
六、菌體表面疏水性程度之測定 ---------------------------------------------8
第二章 臺灣北部海域真菌附著及其組成 ---------------------------------12
中文摘要 --------------------------------------------------------------------------12
英文摘要 --------------------------------------------------------------------------13
壹、前言 --------------------------------------------------------------------------14
貳、材料與方法 ------------------------------------------------------------------15
一、材料 -----------------------------------------------------------------------15
二、方法 -----------------------------------------------------------------------16
(一)懸吊片與放置架 ------------------------------------------------16
(二)懸吊片處理 ------------------------------------------------------16
(三)懸吊片放置及採樣 ---------------------------------------------16
(四)微生物分析 ------------------------------------------------------17
(五)海水總有機碳及氨氮含量分析 --------------------------------17
(六)真菌之分離與純化 ---------------------------------------------17
(七)真菌之鑑定 ------------------------------------------------------17
參、結果與討論 ------------------------------------------------------------------18
一、海水分析 --------------------------------------------------------------18
二、不同季節真菌附著之比較 ----------------------------------------18
三、不同地點真菌附著之比較 ----------------------------------------19
四、不同材料片真菌附著之比較 --------------------------------------19
五、附著真菌之分離與鑑定 --------------------------------------------20
肆、結論 ---------------------------------------------------------------------------22
第三章、海水環境因子對真菌生長及附著能力之影響 ------------------40
中文摘要 --------------------------------------------------------------------------40
英文摘要 --------------------------------------------------------------------------41
壹、前言 ---------------------------------------------------------------------------42
貳、材料與方法 ------------------------------------------------------------------43
一、材料 --------------------------------------------------------------------43
二、方法 --------------------------------------------------------------------44
(一)菌株活化 ---------------------------------------------------------44
(二)生長及附著測試菌株之篩選 -----------------------------------44
(三)酵母菌生長曲線 ------------------------------------------------44
(四)玻璃片附著菌數之分析 --------------------------------------45
(五)環境因子對真菌生長及附著影響之試驗 --------------------45
壹、 結果與討論 -----------------------------------------------------------------47
一、測試菌株之篩選 -----------------------------------------------------47
二、海水中生長測試 -----------------------------------------------------47
三、碳源對酵母菌的生長及附著之影響 -----------------------------47
四、氮源對酵母菌的生長及附著之影響 -----------------------------48
五、鹽類對酵母菌的生長及附著影響 --------------------------------49
六、海水溫度與pH值對酵母菌生長及附著的影響 ---------------49
肆、結論 ---------------------------------------------------------------------------50
第四章 酵母菌表面疏水性及細菌存在與否對其附著能力之影響 ---61
中文摘要 --------------------------------------------------------------------------61
英文摘要 --------------------------------------------------------------------------62
壹、前言 ---------------------------------------------------------------------------63
貳、材料與方法 ------------------------------------------------------------------64
一、材料 --------------------------------------------------------------------64
二、方法 --------------------------------------------------------------------65
(一)細菌對酵母菌附著之影響 ------------------------------------65
(二)菌體表面疏水性之測定 ---------------------------------------66
(三)菌株於海水中表面疏水性及附著力變化之試驗 -----------67
(四)玻璃片上附著菌數之分析 ------------------------------------67
(五)懸浮液中菌數之分析 ------------------------------------------67
參、結果與討論 ------------------------------------------------------------------68
一、細菌對真菌附著能力之影響 --------------------------------------68
二、菌體表面疏水性對附著之影響 -----------------------------------69
肆、結論 ---------------------------------------------------------------------------71
參考文獻 --------------------------------------------------------------------------79

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