苯甲醯尿素類殺蟲劑二福隆、氟芬隆及諾伐隆在土壤的垂直分佈及對不同土層細菌族群的影響

The movement and distribution of insecticides Diflubenzuron, flufenoxuron and novaluron in soils and the effects on bacterial community of different soil layers

蕭鈺齡(Yu-Ling Hsiao)

指導教授 ： 王一雄 ； 顏瑞泓

國立臺灣大學/生物資源暨農學院/農業化學研究所/碩士(2008年)

https://doi.org/10.6342/NTU.2008.01221

摘要

本研究中利用土壤管柱淋洗試驗針對三種苯甲醯尿素類殺蟲劑：二福隆 (Diflubenzuron)、氟芬隆 (Flufenoxuron)及諾伐隆 (Novaluron) 在土壤中的垂直移動及淋洗過程中，土壤細菌族群結構改變情形進行探討。二福隆在國內常用於田間害蟲的防治，例如：毛豆、茶樹，或是居家防治蟑螂、蚊子；氟芬隆則為豆類、薑的蟲害推薦用藥；諾伐隆是近三年內才在國內公告的殺蟲劑，主要用於蕃茄夜蛾及玉米螟的防治。供試土壤選用酸性及微鹼性的二種土壤，分別來自桃園農業改良場 (Pu) 及花蓮農業改良場 (Wl)，其質地分別為壤土及砂質壤土，pH為5.06及7.62。本實驗於管柱上方加入65 g含50 mg/kg殺蟲劑之風乾土壤，在室溫下 (30℃) 進行淋洗，收集淋洗液，以HPLC分析其中農藥含量。淋洗結束後，分析土壤管柱不同深度之農藥殘量，同時進行土壤總DNA的萃取，再使用聚合酶鏈鎖反應 (Polymerase chain reaction, PCR) 擴增微生物族群的16S rRNA片段，配合變性梯度凝膠電泳 (denaturing gradient gel electrophoresis, DGGE) 觀察不同處理下各個深度土壤中的細菌族群結構。
結果顯示三種苯甲醯尿素類殺蟲劑二福隆、氟芬隆及諾伐隆在兩種土壤中經過10週的淋洗後，皆在0~5公分處有最大的殘留量，其在Pu土壤中的殘留量分別為0.15%、37.06%、40.79%；在Wl土壤中的殘留量則為0.32%、63.34%、54.51%。而在5~10公分處則有少量的殺蟲劑殘留，由此可知三種苯甲醯尿素殺蟲劑在兩種土壤中都不易隨淋洗液發生垂直移動。
在不同處理的土壤管柱中的細菌族群結構變化情形，未添加殺蟲劑之空白試驗中可發現，經過10週的淋洗後，隨著土壤管柱深度增加，菌群結構會發生變化，推測淋洗可能會造成菌群的改變。而在添加殺蟲劑的處理中，從PCR-DGGE指紋圖譜可看出，添加殺蟲劑後可能造成土壤中某些優勢菌群的生長，並導致部分細菌族群消失。經過UPGMA方法進行聚類分析後，發現在Wl土壤管柱第一層之菌群和其他深度的菌群比較相似度較低，則和土壤管柱淋洗試驗中殺蟲劑主要分布於管柱0~5公分處的結果相符合，由此可推測，添加殺蟲劑對於Wl土壤細菌族群結構會造成一定的影響；Pu土壤中細菌族群則可能受淋洗或添加殺蟲劑影響，環境有劇烈的變動而使細菌族群結構改變。

關鍵字

二福隆 ； 氟芬隆 ； 諾伐隆 ； 變性梯度凝膠電泳 ； 淋洗

並列摘要

The movement and distribution of benzoylurea insecticides diflubenzuron, flufenoxuron and novaluron in soils and the effects on bacterial community of different soil layers were studied in this research. Diflubenzuron was usually used for prevention and cure of the pests in the field, or in the house. Flufenoxuron recommended for the pests of legumes and ginger. However novaluron was a pesticide that licensed in these three years in Taiwan, mainly prevents pests of tomatoes and corns. Two soils used in this study were sampled from Taoyuan District Agricultural Research and Extension Station and Hualien District Agricultural Research and Extension Station. The soil (Pu) from Taoyuan was loam, pH 5.06 and the other one (Wl) from Hualien was sandy loam, pH 7.62. In this research, dried soil with 50 mg/kg pesticides was added on the top of the soil columns. The soil columns were leached at the room temperature (30℃), and the leachates were collected and analyzed by HPLC (high-performance liquid chromatography) for the pesticides. After 10 weeks leaching, soil columns were analyzed for the residues of pesticides layer by layer; meanwhile the DNA of the total soil microorganisms extracted for the next step in the procedure for the experiment was PCR-DGGE (Polymerase chain reaction- denaturing gradient gel electrophoresis). The variation of bacterial communities was performing with counting the 16S rDNA band on DGGE patterns. The results showed that after leaching periods the pesticides diflubenzuron, flufenoxuron and novaluron hardly moved downward in the soil columns, and mainly appeared in the 0~5 cm depth of columns. The residues in Pu soil columns were 0.15%, 37.06%, and 40.79%; in Wl soil columns were 0.32%, 63.34%, and 54.51%. The DGGE fingerprints showed variations in bacterial communities accompanied depth of soil for blank columns. However, when it comes to the columns treated with pesticides, the DGGE fingerprints indicated that some bacterial communities would be induced, some would be inhibited, others would be inaccessible to the pesticides. Performing Cluster analysis by unweighted pairwise grouping method with mathematical averages (UPGMA) of bacterial community structures with the Wl soil columns, the similarity of the bacterial communities in the 0~5 cm depth is less than the other depth of the soil columns. It matched with the soil leaching test. According to the results, the soil bacterial communities would be influenced by the pesticides; otherwise the bacterial communities in Pu soil could be affected by the variation of the environments due to leaching or pesticides addition.

並列關鍵字

diflubenzuron ； flufenoxuron ； novaluron ； denaturing gradient gel electrophoresis, DGGE ； leaching

參考文獻

3. 行政院環境保護署。2007。住家殺蟲劑使用手冊。

8. Amann, R. I., W. Ludwig and K. H. Schleifer 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol. Rev. 59: 143-169.

9. Ball, D.F. 1964. Loss-on ignition as estimate of organic matter and organic carbon in non-calcareous soils. J. Soil Sci. 15: 84-92.

10. Boopathy R. 2004. Anaerobic biodegradation of no. 2 diesel fuel in soil: a soil column study. Bioresour. Technol. 94: 143-151.

11. Burrows, L. A. and C. A. Edwards. 2004. The Use of Integrated Soil Microcosms to Assess the Impact of Carbendazim on Soil Ecosystems. Ecotoxicology. 13: 143-161.