Ragavendran C, Srinivasan R, Kim Myunghee, Natarajan Devarajan
Natural Drug Research Laboratory, Department of Biotechnology, School of Biosciences, Periyar University, Salem, India.
Department of Food Science and Technology, College of Life and Applied Sciences, Yeungnam University, Gyeongsan, South Korea.
Front Pharmacol. 2018 Nov 26;9:1355. doi: 10.3389/fphar.2018.01355. eCollection 2018.
Vector-borne diseases like malaria, filariasis, and dengue are transmitted by mosquitoes and they cause global mortality and morbidity due to an increased resistance against commercial insecticides. The present study was aimed to evaluate the neurobehavioral toxicity, knock-down effect, histopathology, ovicidal, adulticidal, and smoke toxicity effect of extract against three mosquito species, namely , and (Diptera: Culicidae). The isolated fungal strain was identified as (GenBank accession no: KX694148.1) through morphological and molecular (phylogenetic) analysis. The morphological changes in the treated fourth instar larvae shown the demelanization of cuticle and shrinkage of the internal cuticle of anal papillae. The time duration of extract exposure against the larvae determines the level of toxicity. The extract treated larvae were displayed excitation, violent vertical and horizontal movements with aggressive anal biting behavior as the toxic effect on the neuromuscular system. The results of the biochemical analysis indicated that a decrease in the level of acetylcholinesterase, α-carboxylesterase, and β-carboxylesterase in extract treated fourth instar larvae of all tested mosquito species. The findings of histopathological investigation shown the disorganization of the abdominal region, mainly in mid, hindgut, and gastric caeca, loss of antenna, lateral hair, caudal hair, upper and lower head hairs in the mycelium extract treated , and . The ovicidal bioassay test results showed the mosquito hatchability percentage was directly related to the concentrations of mycelium extract. Nil hatchability of mosquito eggs was noticed at 500 μg/ml concentration. The adulticidal activity of fungal mycelia ethyl acetate extract resulted in a dose-dependent activity (15 and 30 min recovery periods). The higher concentration of extract (1000 mg/L) acted as a repellent, the adult mosquitoes showed restless movement, uncontrolled/anesthetic flight at last died. The better adulticidal activity was observed in the ethyl acetate extract against followed by with the best score of LD and LD values and nil mortality was found in the control. The results of smoke toxicity assay of the mycelia extract exhibited significant mortality rate against (91%), (89%), and (84%). In addition, the present investigation reported the stability and toxic effects of mycelium ethyl acetate extract on . The swimming speed (0.88 mm s) of was reduced with ethyl extract 24 h treatment whereas, the control showed the normal speed of 2.96 mm s. Altered behavior and swimming movement were observed in the 8 h mycelium extract treated . A pale yellow color substance (metabolites) was found in the mid-gut region of the mycelial extract exposed . The outcome of the present study, suggest that the metabolites might serve as an alternative, cost-effective, eco-friendly, and target specific mosquitocidal agent in the future.
疟疾、丝虫病和登革热等病媒传播疾病由蚊子传播,由于对商业杀虫剂的抗性增强,它们导致了全球范围内的死亡率和发病率上升。本研究旨在评估提取物对三种蚊虫(即致倦库蚊、白纹伊蚊和埃及伊蚊,双翅目:蚊科)的神经行为毒性、击倒效果、组织病理学、杀卵、杀成虫和烟雾毒性作用。通过形态学和分子(系统发育)分析,将分离出的真菌菌株鉴定为球孢白僵菌(GenBank登录号:KX694148.1)。经处理的四龄幼虫的形态变化表现为表皮脱黑色素沉着以及肛乳头内表皮收缩。提取物对幼虫的暴露持续时间决定了毒性水平。经提取物处理的幼虫表现出兴奋、剧烈的垂直和水平运动以及具有攻击性的肛门叮咬行为,这是对神经肌肉系统的毒性作用。生化分析结果表明,在所有测试蚊虫物种经提取物处理的四龄幼虫中,乙酰胆碱酯酶、α - 羧酸酯酶和β - 羧酸酯酶水平均有所下降。组织病理学研究结果显示,在经菌丝体提取物处理的致倦库蚊、白纹伊蚊和埃及伊蚊中,腹部区域出现紊乱,主要集中在中肠、后肠和胃盲囊,触角、侧毛、尾毛、头部上下毛缺失。杀卵生物测定试验结果表明,蚊虫孵化率与菌丝体提取物浓度直接相关。在500μg/ml浓度下,未观察到蚊卵孵化。真菌菌丝体乙酸乙酯提取物的杀成虫活性呈现剂量依赖性(恢复期为15和30分钟)。提取物的较高浓度(1000mg/L)起到驱避作用,成年蚊子表现出不安的运动、不受控制的/麻醉性飞行,最终死亡。乙酸乙酯提取物对致倦库蚊的杀成虫活性表现更佳,其次是白纹伊蚊,其LD50和LD90值得分最佳,而对照组未出现死亡情况。菌丝体提取物的烟雾毒性试验结果显示,对致倦库蚊(91%)、白纹伊蚊(89%)和埃及伊蚊(84%)有显著死亡率。此外,本研究报告了球孢白僵菌菌丝体乙酸乙酯提取物对斑马鱼的稳定性和毒性作用。经乙酸乙酯提取物处理24小时后,斑马鱼的游泳速度(0.88mm/s)降低,而对照组的正常速度为2.96mm/s。在经8小时球孢白僵菌菌丝体提取物处理的斑马鱼中观察到行为和游泳运动的改变。在暴露于菌丝体提取物的斑马鱼中肠区域发现一种淡黄色物质(代谢产物)。本研究结果表明,球孢白僵菌代谢产物未来可能成为一种替代的、具有成本效益、生态友好且靶向特定的杀蚊剂。