Mohammed Youssef M M, Badawy Mohammed E I
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.
J Environ Sci Health B. 2017 Oct 3;52(10):752-761. doi: 10.1080/03601234.2017.1356666. Epub 2017 Sep 22.
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.
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。