Optimization of Growth Response Parameters, Screening and Molecular Detection of Pesticide Degradation Genes in Bacterial Isolates from Agricultural Soils

Anas, A., Shamsudeen, U., Ibrahim, Y.
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Abstract

Study’s Novelty/ Excerpt This study demonstrated the capability of bacteria isolated from farmland soils in Kano Metropolis to biodegrade dichlorvos and carbofuran pesticides, identifying Bacillus sp., Serratia sp., and Pseudomonas sp. as key degraders. Optimization experiments revealed that Serratia sp. thrived at 100 mg/L dichlorvos concentration and 35°C, while Pseudomonas sp. showed maximum growth at 300 mg/L carbofuran concentration and 30°C, both with a pH of 7.0, 100 rpm agitation, and 5-day incubation period. Molecular analysis confirmed the presence of opd and mcd genes in Serratia sp. and Pseudomonas sp., respectively, highlighting their potential for effective bioremediation of pesticide-contaminated soils. Full Abstract Pesticides are organic compounds synthesized and used for pest control. The excessive and continuous dispersion of pesticides in the environment results in environmental pollution, necessitating remediation. This study investigated the potential of bacteria isolated from farmland soils in Kano Metropolis, Kano State, Nigeria, with a history of dichlorvos (2,2-dichlorovinyldimethylphosphate) and carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl methylcarbamate) application, to biodegrade these pesticides. Three sampling sites were involved in sample collection, and the soil physicochemical parameters from each sample were determined. Isolation, identification, and screening of the bacterial isolates capable of utilizing the pesticides as sole sources of carbon were carried out. The following parameters (concentration of the pesticides, pH, temperature, agitation, and incubation time) were optimized to maximize degradation. The potent bacterial isolates were further subjected to molecular analysis for the detection of opd and mcd genes. The pesticide-degrading bacteria were identified as Bacillus sp., Serratia sp., and Pseudomonas sp. Serratia sp. recorded the highest growth in the presence of 1% v/v dichlorvos, while Pseudomonas sp. exhibited maximum growth at a 1% w/v carbofuran concentration. The optimized conditions that yielded the maximum microbial growth are: 100 mg/L pesticide concentration for Serratia sp and 300 mg/L for Pseudomonas sp, a pH of 7.0 and an agitation level of 100 rpm for both organisms, a temperature of 35°C for Serratia sp and 30°C for Pseudomonas sp, and an incubation time of 5 days for both organisms. The opd and mcd genes were identified from Serratia sp. and Pseudomonas sp. respectively. These results suggest that the isolated bacteria have the potential to degrade dichlorvos and carbofuran pesticides from the contaminated soil
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农业土壤中细菌分离物的生长响应参数优化、农药降解基因的筛选和分子检测
研究的新颖性/摘要本研究证明了从卡诺市农田土壤中分离的细菌生物降解敌敌畏和呋喃丹农药的能力,确定芽孢杆菌、沙雷氏菌和假单胞菌为主要降解菌、优化实验表明,沙雷氏菌在敌敌畏浓度为 100 毫克/升、温度为 35 摄氏度的条件下生长旺盛,而假单胞菌在呋喃丹浓度为 300 毫克/升、温度为 30 摄氏度的条件下生长旺盛,两者的 pH 值均为 7.0,搅拌转速均为 100 转/分,培养期均为 5 天、分子分析证实,沙雷氏菌和假单胞菌分别含有 opd 和 mcd 基因,这表明它们具有对受农药污染的土壤进行有效生物修复的潜力。农药在环境中的过量和持续散布造成了环境污染,因此有必要采取补救措施。本研究调查了从尼日利亚卡诺州卡诺市农田土壤中分离出的细菌降解这些农药的潜力,这些土壤曾使用过敌敌畏(2,2-二氯乙烯基二甲基磷酸酯)和克百威(2,3-二氢-2,2-二甲基-7-苯并呋喃基甲基氨基甲酸酯)。研究人员在三个采样点采集了样品,并测定了每个样品的土壤理化参数。对能够利用农药作为唯一碳源的细菌进行了分离、鉴定和筛选。对以下参数(农药浓度、pH 值、温度、搅拌和培养时间)进行了优化,以最大限度地提高降解效果。对分离出的强效细菌进一步进行分子分析,以检测 opd 和 mcd 基因。在 1%(体积分数)敌敌畏浓度下,沙雷氏菌的生长速度最快,而在 1%(体积分数)呋喃丹浓度下,假单胞菌的生长速度最快。微生物生长量最大的优化条件是沙雷氏菌的杀虫剂浓度为 100 毫克/升,假单胞菌的杀虫剂浓度为 300 毫克/升;两种生物的 pH 值均为 7.0,搅拌水平均为 100 转/分;沙雷氏菌的温度为 35 摄氏度,假单胞菌的温度为 30 摄氏度;两种生物的培养时间均为 5 天。分别从沙雷氏菌和假单胞菌中鉴定出了 opd 和 mcd 基因。这些结果表明,分离出的细菌具有降解受污染土壤中敌敌畏和呋喃丹农药的潜力。
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