環保專案成果報告查詢系統

專案計畫編號	EPA-98-1401-02-01
經費年度	098
計畫經費	490 千元
專案開始日期	2009/03/12
專案結束日期	2009/10/30
專案主持人	劉秀美
主辦單位	環檢所
承辦人
執行單位	國立台灣海洋大學
專案分類	環境監測及檢測
中文關鍵字	原油、去氫酶活性、脂溶性肥料、最確數方法
英文關鍵字	Crude oil、Dehydrogenase activity、Lipophilic fertilizers、The Most Probable Number Method
協同主持人
共同主持人
計畫聯絡人	劉秀美
計畫聯絡電話	02-24622192#5307
計畫聯絡信箱	smliu@mail.ntou.edu.tw

中文摘要	於此計畫我們將石門鄉油污染海岸劃定一污染試驗區 (經度121°33'36" E、緯度 25°15'56"N) ，將其分為潮汐區與非每月滿潮潮汐區，此二區又再細分為添加肥料區與非添加肥料區。選用一脂溶性氮磷肥料，每月定期噴灑此肥料於上述二添加肥料區。也定期追蹤這四個試驗區岩石上的藻類生長速率，以及岩石上沉積物與底沙中原油被分解情形，我們也追蹤岩石上沉積物與底沙中氨氮、硝酸氮、總磷濃度與其中微生物去氫酶酵素活性、原油分解菌菌數之變化以及其菌相。研究發現在原油污染海岸岩石上藻類之覆蓋率除了潮汐添加肥料區較不顯著外，其他三區之覆蓋率主要有兩次高峰。不過這些藻類的生長與原油(碳氫化合物)之分解或是肥料的添加似無關聯性。雖然岩石上沉積物與底沙中逐月氨氮、硝酸氮、總磷濃度無明顯趨勢變化，不過其中之油污是逐漸下降，添加肥料區油污濃度下降則較快。而非每月滿潮潮汐區添加肥料區在6月初，潮汐添加肥料區在7月初原油分解菌之菌數達最高點。非每月滿潮潮汐非添加肥料區與潮夕非添加肥料區，菌數無顯著的最高點。可是從7月11日到8月6日間，各區菌數都降低很多。底沙或沉積物中微生物去氫酶活性，不論在潮汐與非每月滿潮潮汐區中、添加肥料區或是非添加肥料區其去氫酶活性，在5月11日至7月11日間都有增加之趨勢，添加肥料區有較高的去氫酶活性。同樣地7月11日至8月6日間各區去氫酶活性皆下降。可能7月11日到8月6日間，環境中的原油量下降，再加上氣溫炎熱造成菌數與去氫酶活性的降低。因此綜觀此研究結果，添加肥料的確增加原油分解菌之菌數與去氫酶活性，也加速原油之分解。以Bushnell-Haas broth培養基，利用MPN方法篩選出耐鹽且具原油分解功能的微生物，並比較每月滿潮潮汐區與非每月滿潮潮汐區的微生物菌相差異，及添加肥料與非添加肥料微生物菌相差異。結果發現潮汐區與非潮汐區之微生物菌相差異不大。添加肥料後珊瑚礁岩沉積物之微生物種類增加，底沙則差異不大。從原油污染之底沙與珊瑚礁岩沉積物中共篩選出15種菌株，其中柴油食烷菌（Alcanivorax dieselolei）、威尼斯不動桿菌（Acinetobacter venetianu）、堅強芽胞桿菌（Bacillus firmus）、維斯假絲酵母菌（Candida viswanathii）、瑪麗絲迪茨菌（Dietzia maris）、新鞘氨醇單胞菌（Novosphingobium pentaromativorans）、類產鹼假單胞菌（Pseudomonas pseudoalcaligenes）、施氏假單胞菌（P. stutzeri）、移動醋桿菌（Tistrella mobilis）、纖維化纖維菌（Cellulosimicrobium cellulans）、碘短桿菌（Brevibacterium iodinum）等11株具油分解能力。其餘4株愛希顏氏海洋桿菌（Oceanbacillus iheyensis）、美麗鹽單胞菌（Halomonas venusta）、海藻希瓦氏菌（Shewanella algae）、腐生葡萄球菌（Staphylococcus saprophyticus）等不具原油分解能力，為常見海洋魚類微生物。另經文獻查詢及測試發現稀有芽胞桿菌（Bacillus pumilus）及枯草芽胞桿菌（Bacillus subtilis）等2種菌株可耐乾旱、抗UV且具原油分解之能力。本研究嘗試利用篩選出之15種菌株應用於生物復育整治實驗，發現油污染海岸現地直接添加菌種效果不顯著，但實驗室外加菌種翻堆方式，則有顯著效果，可能是現地試驗易受到氣象天候（如颱風）、潮汐及周圍環境的影響，導致效果不佳。未來可將菌株製成混合製劑用於現地復育，期有助於原油污染海岸油污之清除。
中文關鍵字	原油、去氫酶活性、脂溶性肥料、最確數方法

中文摘要

於此計畫我們將石門鄉油污染海岸劃定一污染試驗區 (經度121°33'36" E、緯度 25°15'56"N) ，將其分為潮汐區與非每月滿潮潮汐區，此二區又再細分為添加肥料區與非添加肥料區。選用一脂溶性氮磷肥料，每月定期噴灑此肥料於上述二添加肥料區。也定期追蹤這四個試驗區岩石上的藻類生長速率，以及岩石上沉積物與底沙中原油被分解情形，我們也追蹤岩石上沉積物與底沙中氨氮、硝酸氮、總磷濃度與其中微生物去氫酶酵素活性、原油分解菌菌數之變化以及其菌相。研究發現在原油污染海岸岩石上藻類之覆蓋率除了潮汐添加肥料區較不顯著外，其他三區之覆蓋率主要有兩次高峰。不過這些藻類的生長與原油(碳氫化合物)之分解或是肥料的添加似無關聯性。雖然岩石上沉積物與底沙中逐月氨氮、硝酸氮、總磷濃度無明顯趨勢變化，不過其中之油污是逐漸下降，添加肥料區油污濃度下降則較快。而非每月滿潮潮汐區添加肥料區在6月初，潮汐添加肥料區在7月初原油分解菌之菌數達最高點。非每月滿潮潮汐非添加肥料區與潮夕非添加肥料區，菌數無顯著的最高點。可是從7月11日到8月6日間，各區菌數都降低很多。底沙或沉積物中微生物去氫酶活性，不論在潮汐與非每月滿潮潮汐區中、添加肥料區或是非添加肥料區其去氫酶活性，在5月11日至7月11日間都有增加之趨勢，添加肥料區有較高的去氫酶活性。同樣地7月11日至8月6日間各區去氫酶活性皆下降。可能7月11日到8月6日間，環境中的原油量下降，再加上氣溫炎熱造成菌數與去氫酶活性的降低。因此綜觀此研究結果，添加肥料的確增加原油分解菌之菌數與去氫酶活性，也加速原油之分解。以Bushnell-Haas broth培養基，利用MPN方法篩選出耐鹽且具原油分解功能的微生物，並比較每月滿潮潮汐區與非每月滿潮潮汐區的微生物菌相差異，及添加肥料與非添加肥料微生物菌相差異。結果發現潮汐區與非潮汐區之微生物菌相差異不大。添加肥料後珊瑚礁岩沉積物之微生物種類增加，底沙則差異不大。從原油污染之底沙與珊瑚礁岩沉積物中共篩選出15種菌株，其中柴油食烷菌（Alcanivorax dieselolei）、威尼斯不動桿菌（Acinetobacter venetianu）、堅強芽胞桿菌（Bacillus firmus）、維斯假絲酵母菌（Candida viswanathii）、瑪麗絲迪茨菌（Dietzia maris）、新鞘氨醇單胞菌（Novosphingobium pentaromativorans）、類產鹼假單胞菌（Pseudomonas pseudoalcaligenes）、施氏假單胞菌（P. stutzeri）、移動醋桿菌（Tistrella mobilis）、纖維化纖維菌（Cellulosimicrobium cellulans）、碘短桿菌（Brevibacterium iodinum）等11株具油分解能力。其餘4株愛希顏氏海洋桿菌（Oceanbacillus iheyensis）、美麗鹽單胞菌（Halomonas venusta）、海藻希瓦氏菌（Shewanella algae）、腐生葡萄球菌（Staphylococcus saprophyticus）等不具原油分解能力，為常見海洋魚類微生物。另經文獻查詢及測試發現稀有芽胞桿菌（Bacillus pumilus）及枯草芽胞桿菌（Bacillus subtilis）等2種菌株可耐乾旱、抗UV且具原油分解之能力。本研究嘗試利用篩選出之15種菌株應用於生物復育整治實驗，發現油污染海岸現地直接添加菌種效果不顯著，但實驗室外加菌種翻堆方式，則有顯著效果，可能是現地試驗易受到氣象天候（如颱風）、潮汐及周圍環境的影響，導致效果不佳。未來可將菌株製成混合製劑用於現地復育，期有助於原油污染海岸油污之清除。

中文關鍵字

原油、去氫酶活性、脂溶性肥料、最確數方法

類型	檔名	檔案大小	說明	最新版公開日期
期末報告	Microsoft Word - EPA-98-1401-02-01.pdf	1MB		2010/3/1 上午 12:00:00

摘要	The aim of this study was to examine the effects of lipophilic fertilizers (uric acid and lecithin) on the degradation of crude oil hydrocarbons in the oil-spilled coast. An area of oil-spilled coast was selected and dived into tidal and non tidal zones. These zones were further divided into fertilizer amendment and non-amendment zones. Lipophilic fertilizers were spread into fertilizer amendment zones monthly. Degradation of crude oil in these test zones were investigated by measuring concentration of the total hydrocarbon, NH4+-N, NO3--N, total phosphorus, number of petroleum degraders, and microbial dehydrogenase activity in sands or sediments in these zones monthly. Although there were no any trend in the concentration of NH4+-N, NO3--N, and total phosphorus, concentration of total hydrocarbon in these zones decreased during the test period. The number of petroleum degrader reached the highest in the beginning of June in non tidal-fertilizer amendment zone, while in the beginning of July in the tidal-fertilizer amendment zones. The number of petroleum degraders did not change much in non-fertilizer amendment zones. The number of petroleum degraders in these zones all decreased sharply after July. Dehydrogenase activity increased from May to July and then decreased in sand/sediment of all these zones. However, dehydrogenase activity in sand/sediment was much higher in fertilizer-amended than in fertilizer-non amended zones. Based on these results, we know that fertilizer amendment increased the number of petroleum degraders and microbial activity, and enhanced crude oil removal. In this study, we also used Bushnell-Haas broth to enrich and isolate petroleum degraders from sand/sediment of these oil-spilled zones. We found there was not much difference in the microbial community of tidal and non tidal zones. Amendment of fertilizer increased species of petroleum degraders in sediments but not in sands. We isolated 15 strains. 11 strains, e.g. Alcanivorax dieselolei, Acinetobacter venetianu, Bacillus firmus, Candida viswanathii, Dietzia maris, Novosphingobium pentaromativorans, Pseudomonas pseudoalcaligenes, P. stutzeri, Tistrella mobilis, Cellulosimicrobium cellulans, Brevibacterium iodinum can degrade crude oil. Four strains, e.g. Oceanbacillus iheyensis, Halomonas venusta, Shewanella algae, Staphylococcus saprophyticus can not degrade crude oil. However, using consortia of these 11 petroleum-degrading bacteria for on site bioremediation did not get good results. Various environmental factors such as weather, tidal might affect these results. Further works are needed to prepare better consortia for bioremediation.
英文關鍵字	Crude oil、Dehydrogenase activity、Lipophilic fertilizers、The Most Probable Number Method

摘要

The aim of this study was to examine the effects of lipophilic fertilizers (uric acid and lecithin) on the degradation of crude oil hydrocarbons in the oil-spilled coast. An area of oil-spilled coast was selected and dived into tidal and non tidal zones. These zones were further divided into fertilizer amendment and non-amendment zones. Lipophilic fertilizers were spread into fertilizer amendment zones monthly. Degradation of crude oil in these test zones were investigated by measuring concentration of the total hydrocarbon, NH4+-N, NO3--N, total phosphorus, number of petroleum degraders, and microbial dehydrogenase activity in sands or sediments in these zones monthly. Although there were no any trend in the concentration of NH4+-N, NO3--N, and total phosphorus, concentration of total hydrocarbon in these zones decreased during the test period. The number of petroleum degrader reached the highest in the beginning of June in non tidal-fertilizer amendment zone, while in the beginning of July in the tidal-fertilizer amendment zones. The number of petroleum degraders did not change much in non-fertilizer amendment zones. The number of petroleum degraders in these zones all decreased sharply after July. Dehydrogenase activity increased from May to July and then decreased in sand/sediment of all these zones. However, dehydrogenase activity in sand/sediment was much higher in fertilizer-amended than in fertilizer-non amended zones. Based on these results, we know that fertilizer amendment increased the number of petroleum degraders and microbial activity, and enhanced crude oil removal. In this study, we also used Bushnell-Haas broth to enrich and isolate petroleum degraders from sand/sediment of these oil-spilled zones. We found there was not much difference in the microbial community of tidal and non tidal zones. Amendment of fertilizer increased species of petroleum degraders in sediments but not in sands. We isolated 15 strains. 11 strains, e.g. Alcanivorax dieselolei, Acinetobacter venetianu, Bacillus firmus, Candida viswanathii, Dietzia maris, Novosphingobium pentaromativorans, Pseudomonas pseudoalcaligenes, P. stutzeri, Tistrella mobilis, Cellulosimicrobium cellulans, Brevibacterium iodinum can degrade crude oil. Four strains, e.g. Oceanbacillus iheyensis, Halomonas venusta, Shewanella algae, Staphylococcus saprophyticus can not degrade crude oil. However, using consortia of these 11 petroleum-degrading bacteria for on site bioremediation did not get good results. Various environmental factors such as weather, tidal might affect these results. Further works are needed to prepare better consortia for bioremediation.

英文關鍵字

Crude oil、Dehydrogenase activity、Lipophilic fertilizers、The Most Probable Number Method