Molecular Identification of Leptospira interrogans and Its Treatment with Andrographis paniculata Extract

Pub Date : 2024-03-20 DOI:10.3103/s0891416823040110
Abirami Madhaiyan, V. Muthumani, E. Angel Jemima, K. Rubika, V. Bharathi, N. Parameswari, V. Kowsalya Devi, K. Ramesh, P. Poongothai
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Abstract

Background: Leptospirosis is a zoonotic disease that transmitted to humans by rodents and domestic animals that infected by Leptospira bacteria and considered as a endemic disease in South India and North-East India. Andrographis paniculata is an ancient herb with various therapeutic properties and has Andrographolide bioactive compound. Purpose: The aim of the present study is to isolate and molecular identification by ITS sequencing of the Leptospira interrogans from the soil sample of paddy field and to analyse the anti-leptospirosis activity of Andrographis paniculata extract in in vitro. Methods: Andrographis paniculata extract was extracted using methanol and screening of phytocompound was done by GC-MS analysis. Leptospira species was isolated from the soil sample and molecular identification was performed by 16s rRNA gene sequencing. BLAST and phylogenetic tree analysis confirmed the presence of Leptospira interrogans. Molecular Docking analysis was performed to study the interaction between the Andrographolide and Leptospira LRR protein. Anti-leptospirosis activities of Andrographis paniculata extracted using different solvents were determined by Minimum Inhibitory Concentration (MIC) methods. Results: The 16s rRNA sequencing and BLAST results confirmed the presence of Leptospirosis interrogans. The phylogenetic tree analysis showed the distance between the Leptospirosis species. Furthermore, the GCMS analysis of Andrographis paniculata methanolic extract showed the presence of phytocompound Andrographolide. Molecular docking analysis of Andrographolide with the LRR protein showed stable binding. The in vitro anti-leptospiral activity of different Andrographis paniculata extract showed the maximum inhibition at 500 μg/mL and the maximum MIC was observed at methanolic extract of A. paniculata at 31.2 μg/mL with 64.65 ± 7.33% of inhibition. Conclusion: From our study we concluded that Andrographis paniculata extracted using methanol has good anti-leptospirosis activity and Andrographolide will act as a good therapeutic agent against leptospirosis.

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钩端螺旋体的分子鉴定及穿心莲提取物对其的治疗
背景:钩端螺旋体病是一种人畜共患病,由啮齿动物和家畜感染钩端螺旋体细菌后传染给人类,被认为是印度南部和东北部的地方病。穿心莲是一种古老的草药,具有多种治疗功效,并含有穿心莲内酯生物活性化合物。目的:本研究旨在通过 ITS 测序从稻田土壤样本中分离和分子鉴定钩端螺旋体,并分析穿心莲提取物的体外抗钩端螺旋体病活性。方法:穿心莲提取物用甲醇提取穿心莲提取物,并通过气相色谱-质谱分析筛选植物化合物。从土壤样本中分离出钩端螺旋体,并通过 16s rRNA 基因测序进行分子鉴定。BLAST 和系统树分析证实了钩端螺旋体的存在。进行了分子对接分析,以研究穿心莲内酯与钩端螺旋体 LRR 蛋白之间的相互作用。采用最小抑菌浓度法测定了不同溶剂萃取的穿心莲内酯的抗钩端螺旋体活性。结果:16s rRNA 测序和 BLAST 结果证实了审讯钩端螺旋体的存在。系统发生树分析表明钩端螺旋体物种之间存在距离。此外,穿心莲甲醇提取物的气相色谱分析表明存在植物化合物穿心莲内酯。穿心莲内酯与 LRR 蛋白的分子对接分析表明其结合稳定。不同穿心莲提取物的体外抗钩端螺旋体活性显示,在 500 μg/mL 时抑制作用最大,穿心莲甲醇提取物的最大 MIC 值为 31.2 μg/mL,抑制率为 64.65 ± 7.33%。结论通过研究,我们得出结论:用甲醇提取的穿心莲具有良好的抗钩端螺旋体病活性,穿心莲内酯将成为一种良好的钩端螺旋体病治疗剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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