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分离出的废水真菌土曲霉YESM3对吡虫啉在液体培养基中的生物降解作用。

Biodegradation of imidacloprid in liquid media by an isolated wastewater fungus Aspergillus terreus YESM3.

作者信息

Mohammed Youssef M M, Badawy Mohammed E I

机构信息

a Department of Botany and Microbiology, Faculty of Science , Damanhour University , Damanhour , Egypt.

b Department of Pesticide Chemistry and Technology, Faculty of Agriculture , Alexandria University , Alexandria , Egypt.

出版信息

J Environ Sci Health B. 2017 Oct 3;52(10):752-761. doi: 10.1080/03601234.2017.1356666. Epub 2017 Sep 22.

Abstract

In the present study, a new fungal strain capable of imidacloprid degradation was isolated from agricultural wastewater drain. The fungal strain of YESM3 was identified as Aspergillus terreus based on ITS1-5.8S rDNA-ITS2 gene sequence by PCR amplification of a 500 bp sequence. Screening of A. terreus YESM3 to the insecticide imidacloprid tolerance was achieved by growing fungus in Czapek Dox agar for 6 days at 28°C. High values (1.13 and 0.94 cm cm) of tolerance index (TI) were recorded at 25 and 50 mg L of imidacloprid, respectively in the presence and absence of sucrose. However, at 400 mg L the fungus did not grow. Effects of the imidacloprid concentration, pH, and inoculum size on the biodegradation percentage were tested using Box-Behnken statistical design and the biodegradation was monitored by HPLC analysis at different time intervals. Box-Behnken results indicated that optimal conditions for biodegradation were at pH 4 and two fungal discs (10 mm diameter) in the presence of 61.2 mg L of imidacloprid. A. terreus YESM3 strain was capable of degrading 85% of imidacloprid 25 mg L in Czapek Dox broth medium at pH 4 and 28°C for 6 days under static conditions. In addition, after 20 days of inoculation, biodegradation recorded 96.23% of 25 mg L imidacloprid. Degradation kinetics showed that the imidacloprid followed the first order kinetics with half-life (t) of 1.532 day. Intermediate product identified as 6-chloronicotinic acid (6CNA) as one of the major metabolites during degradation of imidacloprid by using HPLC. Thus, A. terreus YESM3 showed a potential to reduce pollution by pesticides and toxicity in the effected environment. However, further studies should be conducted to understand the biodegradation mechanism of this pesticide in liquid media.

摘要

在本研究中,从农业废水排放口分离出一种能够降解吡虫啉的新真菌菌株。通过PCR扩增500bp序列,基于ITS1-5.8S rDNA-ITS2基因序列将YESM3真菌菌株鉴定为土曲霉。通过在28°C下于查氏琼脂培养基中培养真菌6天,实现了土曲霉YESM3对杀虫剂吡虫啉耐受性的筛选。在有和没有蔗糖的情况下,分别在25mg/L和50mg/L的吡虫啉浓度下记录到较高的耐受指数(TI)值(分别为1.13和0.94cm/cm)。然而,在400mg/L时真菌未生长。使用Box-Behnken统计设计测试了吡虫啉浓度、pH值和接种量对生物降解率的影响,并通过不同时间间隔的HPLC分析监测生物降解情况。Box-Behnken结果表明,生物降解的最佳条件是在pH值为4且存在61.2mg/L吡虫啉的情况下使用两个真菌圆盘(直径10mm)。在静态条件下,土曲霉YESM3菌株能够在pH值为4、28°C的查氏肉汤培养基中于6天内降解25mg/L吡虫啉的85%。此外,接种20天后,生物降解率达到25mg/L吡虫啉的96.23%。降解动力学表明,吡虫啉遵循一级动力学,半衰期(t)为1.532天。通过HPLC鉴定出中间产物6-氯烟酸(6CNA)是吡虫啉降解过程中的主要代谢产物之一。因此,土曲霉YESM3显示出在受影响环境中降低农药污染和毒性的潜力。然而,应进一步开展研究以了解该农药在液体介质中的生物降解机制。

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