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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REFERENCES
Levett, P.N., Leptospirosis, Clin. Microbiol. Rev., 2001, vol. 14, no. 2, pp. 296–326. https://doi.org/10.1128/CMR.14.2.296
Cameron, C.E., Zuerner, R.L., Raverty, S., Colegrove, K.M., Norman, S.A., Lambourn, D.M., Jeffries, S.J., and Gulland, F.M., Detection of pathogenic Leptospira bacteria in pinniped populations via PCR and identification of a source of transmission for zoonotic leptospirosis in the marine environment, J. Clin. Microbiol., 2008, vol. 46, no. 5, pp. 1728–1733. https://doi.org/10.1128/JCM.02022-07
Hornsby, R.L., Alt, D.P., and Nally, J.E., Isolation and propagation of leptospires at 37°C directly from the mammalian host, Sci. Rep., 2020, vol. 10, no. 1, pp. 9620–9628. https://doi.org/10.1038/s41598-020-66526-4
Shivakumar, S., Leptospirosis—current scenario in India, in Medicine Update, Association of Physicians of India, 2008, vol. 18, chap. 106, pp. 799–809. https://doi.org/10.13140/2.1.4905.6968
Shukla, S., Mittal, V., Karoli, R., Singh, P., and Singh, A., Leptospirosis in central & eastern Uttar Pradesh, an underreported disease: A prospective cross-sectional study, Indian J. Med. Res., 2022, vol. 155, no. 1, pp. 66–72. https://doi.org/10.4103/ijmr.IJMR_1811_19
Abas, F., Ismail, A., Lajis, N.H., and Shaari, K., Anti-diabetic activity and metabolic changes obese diabetic rats, Molecules., 2016, vol. 21, no. 8, pp. 1026–1036. https://doi.org/10.3390/molecules21081026
Kaushik, S., Dar, L., Kaushik, S., and Yadav, J.P., Identification and characterization of new potent inhibitors of dengue virus NS5 proteinase from Andrographis paniculata supercritical extracts on in animal cell culture and in silico approaches, J. Ethnopharmacol., 2021, vol. 267, p. 113541. https://doi.org/10.1016/j.jep.2020.113541
Paemanee, A., Hitakarun, A., Wintachai, P., Roytrakul, S., and Smith, D.R., A proteomic analysis of the anti-dengue virus activity of andrographolide, Biomed. Pharmacother., 2019, vol. 109, pp. 322–332. https://doi.org/10.1016/j.biopha.2018.10.054
Bell, J.R., A simple way to treat PCR products prior to sequencing using ExoSAP-IT®, Biotechniques, 2008, vol. 44, no. 6, pp. 834–840. https://doi.org/10.2144/000112890
Green, S.J., Venkatramanan, R., and Naqib, A., Deconstructing the polymerase chain reaction: Understanding and correcting bias associated with primer degeneracies and primer-template mismatches, PLoS One, 2015, vol. 10, no. 5, p. e0128122. https://doi.org/10.1371/journal.pone.0128122
Otto, P., Automated high throughput purification of BigDyeTM terminator fluorescent DNA sequencing reactions using Wizard® MagneSilTM paramagnetic particles, J. Assoc. Lab. Autom., 2002, vol. 7, no. 5, pp. 75–79. https://doi.org/10.1016/S1535-5535(04)00222-9
Hall, B.G., Building phylogenetic trees from molecular data with MEGA, Mol. Biol. Evol., 2013, vol. 30, no. 5, pp. 1229–1235. https://doi.org/10.1093/molbev/mst012
Mkumbe, B.S., Pangastuti, A., and Susilowati, A., Phylogenetic analysis based on internal transcribed spacer region (ITS1-5. 8S-ITS2) of Aspergillus niger producing phytase from Indonesia, AIP Conf. Proc., 2018, vol. 2014, no. 1, p. 020015. https://doi.org/10.1063/1.5054419
Thangavel, M., Umavathi, S., Thangam, Y., Thamaraiselvi, A., and Ramamurthy, M., GC-MS analysis and larvicidal activity of Andrographis paniculata (Burm.F) Wall. Ex Nees. against the dengue vector Aedes aegypti (L) (Diptera: Culicidae), Int. J. Curr. Microbiol. Appl. Sci., 2015, vol. 4, no. 7, pp. 392–403.
Jitapunkul, K., Poachanukoon, O., Hannongbua, S., Toochinda, P., and Lawtrakul, L., Simulation study of interactions between two bioactive components from Zingiber cassumunar and 5-lipoxygenase, Cell. Mol. Bioeng., 2018, vol. 11, no. 1, pp. 77–89. https://doi.org/10.1007/s12195-017-0515-6
Macwilliams, M.P. and Liao, M., Luria Broth (LB) and Luria Agar (LA) Media and Their Uses Protocol, American Society for Microbiology, 2006, pp. 7–9.
Zarantonelli, L., Suanes, A., Meny, P., Buroni, F., Nieves, C., Salaberry, X., Briano, C., Ashfield, N., Da Silva Silveira, C., Dutra, F., Easton, C., Fraga, M., Giannitti, F., Hamond, C., et al., Isolation of pathogenic Leptospira strains from naturally infected cattle in Uruguay reveals high serovar diversity, and uncovers a relevant risk for human leptospirosis, PLoS Neglected Trop. Dis., 2018, vol. 12, no. 9, p. e0006694. https://doi.org/10.1371/journal.pntd.0006694
Siddiqui, S., Ameen, F., Rehman, S., Sarwar, T., and Tabish, M., Studying the interaction of drug/ligand with serum albumin, J. Mol. Liq., 2021, vol. 336, p. 116200. https://doi.org/10.1016/j.molliq.2021.116200
Guerroudj, A.R., Boukabcha, N., Benmohammed, A., Dege, N., Belkafouf, N.E.H., Khelloul, N., Djafri, A., and Chouaih, A., Synthesis, crystal structure, vibrational spectral investigation, intermolecular interactions, chemical reactivity, NLO properties and molecular docking analysis on (E)-N-(4-nitrobenzylidene)-3-chlorobenzenamine: A combined experimental and theoretical study, J. Mol. Struct., 2021, vol. 1240, p. 130589. https://doi.org/10.1016/j.molstruc.2021.130589
Saluja, H., Mehanna, A., Panicucci, R., and Atef, E., Hydrogen bonding: Between strengthening the crystal packing and improving solubility of three haloperidol derivatives, Molecules, 2016, vol. 21, no. 6, p. 719. https://doi.org/10.3390/molecules21060719
Nagarajan, P., Jothiraj, M., Asirwatham, A.R., and Alagappan, U., In vitro anti leptospiral activity of chloroform extract of Piper betle L, World J. Pharm. Sci., 2014, vol. 2, no. 8, pp. 711–715. https://doi.org/10.13140/2.1.1209.1527
ACKNOWLEDGMENTS
The authors are grateful to Trichy Research Institute of Biotechnology PVT for providing the research facilities and technical support to carrying out the research work. We would like to express our sincere gratitude to Dr. E. Angel Jemima for her contributions, continuous support, insightful comments and suggestions to complete the study.
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Madhaiyan, A., Muthumani, V., Jemima, E.A. et al. Molecular Identification of Leptospira interrogans and Its Treatment with Andrographis paniculata Extract. Mol. Genet. Microbiol. Virol. 38, 277–286 (2023). https://doi.org/10.3103/S0891416823040110
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DOI: https://doi.org/10.3103/S0891416823040110