Pallavi Rawat Vidya Chauhan Juhi Chaudhary Chhaya Singh Neha Chauhan


Most bacteria are becoming resistant to almost all of the currently recommended drugs, leading to difficulty in their treatment. The present study focused on evaluating the therapeutic potential of Piper longum fruit extracts in terms of bactericidal, antioxidant and phytochemical evaluation by conducting antibacterial sensitivity tests against four multidrug-resistant Salmonella strains (Salmonella enterica subsp. arizonae, Salmonella Newport, Salmonella enterica ser. Typhi, and Salmonella enterica ser. Paratyphi) obtained from the Institute of Microbial Technology (IMTECH), Chandigarh. The Agar Well Diffusion method and the Minimum Inhibitory concentration (MIC) methods were performed to implement the anti-sensitivity test of crude extracts of the plant. The present study showed that the MIC of the P. longum  was between 0.25-0.0625mg/ml, which was lowest in the aqueous extract at 0.5mg/100µl, and the highest in the methanol extract (1mg/100µl). The Minimum Bactericidal Concentration (MBC) was lowest in aqueous (0.5mg/100µl) and highest in methanol plant extract (1mg/100µl). The methanol extract had the maximum antibacterial potency, whereas the aqueous extract had the lowest. The antioxidant capacity of the plant extracts was determined using a DPPH assay. Methanol plant extract revealed the highest antioxidant power (81.92%) and the lowest was found in the aqueous extract (62.84%). The GC-MS approach identified active bioingredients, important botanicals including caryophyllene, eicosane, and piperazine (potent antibacterial agent) as naphthyridine (having antimicrobial, anticancer, and anti-inflammatory activities), among others. The unique aspect of the study was the effectiveness of P. longum against Salmonella strains that are resistant to multiple antibiotics. This suggests that P. longum can be a great source of novel antibacterial compound for the development of herbal formulations.




Antioxidant, DMSO, DPPH, Multidrug-resistant, Piper longum

Akhtar, M.S., Birhanu, G. & Demisse, S. (2014). Antimicrobial activity of Piper nigrum L. and Cassia didymobotyra L. leaf extract on selected foodborne pathogens. Asian Pac. J. Trop. Med. 4(Suppl 2), S911-S919. https://doi.org/10.1016/S2222-1808(14)60757-X
Brundha, M.P. (2015). A comparative study-the role of skin and nerve biopsy in hansen’s disease. J. Pharm. Sci. Res. 7(10), 837-44.
Brundha, M.P., Pathmashri, V.P. & Sundari, S. (2019). Quantitative changes of red blood cells in cancer patients under palliative radiotherapy-a retrospective study. Res. J. Pharm. Technol. 12(2): 687–92. https://doi.org/10.5958/0974-360X.2019.00122.7
Chauhan, R., Chaudhary, E. & Chauhan, N. (2017). Investigation of Inhibitory Activity and Bioactive Compounds of Piper nigrum Seeds on Selected Human Pathogens. Int. J. Curr. Microbiol. App. Sci. 6(7): 2670-679. https://doi.org/10.20546/ijcmas.2017.607.376
Choudhary, N. & Singh, V. (2018). A census of P. longum’s phytochemicals and their network pharmacological evaluation for identifying novel drug-like molecules against various diseases, with a special focus on neurological disorders. PLoS One. 13(1):e0191006. https://doi.org/10.1371/journal.pone.0191006
do Nascimento, K. F., Moreira, F. M. F., Santos, J. A., Kassuya, C. A. L., Croda, J. H. R., Cardoso, C. A. L., do Carmo Vieira, M., Ruiz, A.L.T.G., Foglio, M.A., de Carvalho, J.E. & Formagio, A. S. N. (2018). Antioxidant, anti-inflammatory, antiproliferative and antimycobacterial activities of the essential oil of Psidium guineense Sw. and spathulenol. Journal of ethnopharmacology, 210: 351-58. https://doi.org/10.1016/j.jep.2017.08.030
Fuchs, P., Loeseken, C., Schubert, J.K. & Miekisch, W. (2010). Breath gas aldehydes as biomarkers of lung cancer. Int. J. Cancer. 126(11): 2663–670. https://doi.org/10.1002/ijc.24970
Go, J., Seo, J.Y., Park, T.S., Ryu, Y.K., Park, H.Y., Noh, J.R., Kim, Y.H, et al. (2018). Piperlongumine activates Sirtuin1 and improves cognitive function in a murine model of Alzheimer’s disease. J. Funct. Foods, 43: 103-11. https://doi.org/10.1016/j.jff.2018.02.002
Gowthami, R., Sharma, N., Pandey, R. & Agarwal, A. (2021). Status and consolidated list of threatened medicinal plants of India. Genet Resour Crop Evol. 68(6): 2235-263. http//doi.org/10.1007/s10722-021-01199-0
Guerrero, T., Caldero´n, D., Zapata, S. & Trueba, G. (2020). Salmonella grows massively and aerobically in chicken fecal matter. Microb Biotechnol. 13: 1678–684. https://doi.org/10.1111/1751-7915.13624
Hwang, I.M., Yang, J.S., Kim, S.H., Jamila, N., Khan, N., Kim, K.S. & Seo, H.Y. (2016). Elemental sea, rock, and bamboo salts are analysed by inductively coupled plasma-optical emission and mass spectrometry. Anal. Lett. 49(17): 2807-821. https://doi.org/10.1080/00032719.20 16.1158831
Jindal, D. & Rani, V. (2022). In Silico Studies of Phytoconstituents from Piper longum and Ocimum sanctum as ACE2 and TMRSS2 Inhibitors: Strategies to Combat COVID-19. Appl Biochem Biotechnol. https://doi.org/10.1007/s12010-022-03827-6
Jubie, S., Ashish, W., Sabaritha, K., Nishanthini, P., Thomas, A., & Antony, J. (2016). Synthesis and In-vitro Anti-Cancer Screening of N^ sup 1^[(Substituted Phenyl) Benzylidene] Benzohydrazides. J. Pharm. Sci & Res. 8(7): 582-85.
Kabir, Y. (2020). Health Benefits of Octacosanol and Other Long-Chain Aliphatic Fatty Alcohols from Plants. Herbal Medicine in India. 413-425. Springer, Singapore. https://doi.org/10.1007/978-981-13-7248-3_25
Khushbu, C., Roshni, S., Anar, P., Carol, M. & Mayuree, P. (2011) Phytochemical and therapeutic potential of Piper longum Linn. a review. Int J Res Ayurveda Pharm 2(1):157–61
Koleva, I.I., van Beek, T.A., Linssen, J.P.H., deGroot, A. & Evstatieva, L.N. (2002). Screening of plant extracts for antioxidant activity: A comparative study on three testing methods. Phytochemical Analysis: Int. J. Plant Chem. Biochem. Techs. 13(1): 8-17. https://doi.org/10.1002/pca.611
Koyama, S., Purk, A., Kaur, M., Soini, H.A., Novotny, M.V., Davis, K., Kao, C.C., Matsunami, H. & Mescher, A. (2019). Beta-caryophyllene enhances wound healing through multiple routes. PloS one. 14(12), e0216104. https://doi.org/10.1371/journal.pone.0216104
Kumar, S., Kamboj, J., Suman. & Sharma, S. (2011). Overview for Various Aspects of the Health Benefits of Piper longum Linn. Fruit. J Acupunct Meridian Stud. 4(2):134-140. https://doi.org/10.1016/S2005-2901(11)6002 0-4
Kumar, S., Kar, A., Beena, C., Patel, J., Sohil, V. & Singh, R. (2021). Antioxidant activities, phenolics, and piperine contents in four Piper species from India. Am. J. Essent. Oil. Nat. Prod. 9(1): 24-31.
Lakhera, S., Devlal, K., Ghosh, A. & Rana, M. (2021). In silico investigation of phytoconstituents of the medicinal herb ‘Piper longum’ against SARS-CoV-2 by molecular docking and molecular dynamics analysis. Results Chem. 3: 100199. https://doi.org/10.1016/j.rechem.2021.100199
Machado, KDC., Paz, M.F.C.J., Oliveira Santos, J.V.D., da Silva, F.C.C., Tchekalarova, J.D., Salehi, B., Islam, M.T., Setzer, W.N., Rad, J.S., Sousa, J.M.D.C. & Cavalcante, A.A.D.C. (2020). Anxiety Therapeutic Interventions of β-Caryophyllene: A Laboratory-Based Study. Nat. Prod. Commun. 15(10), 1934578X20962229. https://doi.org/10.1177/1934578X20962229
McDermott, P.F., Zhao, S. & Tate, H. (2018). Antimicrobial resistance in nontyphoidal Salmonella. Microbiol Spectr. 6. https://doi.org/10.1128/microbiolspec.ARBA-0014-2017.
Mgbeahuruike, E.E., Yrjönen, T., Vuorela, H. & Holm, Y. (2017) Bioactive compounds from medicinal plants: focus on Piper species. S Afr J Bot., 112:54–69. https://doi.org/10.1016/j.sajb.2017.05.007
Moradi, S. & Yousofvand, N. (2016). Peganum Harmala and Piper Longum Plant Rubbing Oil Effect on Pain in Small Male Mice. Biosci. Biotechnol. Res. Asia. 13(2): 821-826. https://dx.doi.org/10.13005/bbra/2102
National Committee for Clinical Laboratory Standards. (1998). Performance Standards for antimicrobial susceptibility testing- eighth informational supplement: Approved Standard M100 S8.
National Committee for Clinical Laboratory Standards. (2000). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Approved Standard, Fifth edition: M7-A5.
Ojha, M., Yadav, D., Kumar, A., Dasgupta, S., & Yadav, R. (2021). 1, 8-naphthyridine derivatives: a privileged scaffold for versatile biological activities. Mini Reviews in Medicinal Chemistry. 21(5): 586-601. https://doi.org/10.21 74/1389557520666201009162804
Radhakrishnan, N., Gnanamani, A. & Mandal, A.B. (2011). A potential antibacterial agent Embelin, a natural benzoquinone extracted from Embelia Ribes. Biol. Med. 3(2): 1-7. https://dx.doi.org/10.1007/s11051-010-9934-1
Rajeshkumar, S. (2016). Anticancer activity of ecofriendly gold nanoparticles against lung and liver cancer cells. J. Genet. Eng. Biotechnol. 14(1): 195–202. https://doi.org/10.1016/j.jgeb.2016.05.007
Reddy, P.S., Jamil, K., Madhusudhan, P., Anjani, G. & Das, B. (2001). Antibacterial activity of isolates from Piper longum and Taxus baccata. Pharm. Biol. 39(3): 236-38. https://doi.org/10.1076/phbi.
Sawhney, S.S., Painuli, R.M. & Chauhan, N. (2011). Evaluation of bactericidal and anticancer properties of fruits of P. longum. Int. J. Pharm. Sci. 3: 282-87.
Sharma, R., Kumari, N., Ashawat, M.S. & Verma, C.P.S. (2020). Standardication and phytochemical screening analysis for herbal extracts: Zingiber officinalis, Rosc., Curcuma longa Linn., Cinnamonum zeylanicum Nees., Piper longum Linn., Boerhaavia diffussa Linn. Asian J. Pharm. Technol. 10(3): 127- 133. https://dx.doi.org/10.5958/2231-5713.2020.00022.7
Smilkov, K., Ackova, D.G., Cvetkovski, A., Ruskovska, T., Vidovic, B. & Atali, M. (2019). Piperine: Old spice and new Nutraceutical? Curr. Pharm. Des. 25(15): 1729-39. https://doi.org/10.2174/1381612825666190701150803
Sultana, N.A., Zilani, M.N.H., Taraq, K.T.M. & Al-Din, M.K. (2019). Phytochemical, antibacterial and antioxidant activity of Piper longum leaves. PharmacologyOnLine. 1, 27-35.
Tack, B., Vanaenrode, J., Verbakel, J.Y., Toelen, J. & Jacobs, J. (2020). Invasive non-typhoidal Salmonella infections in sub-Saharan Africa: A systematic review on antimicrobial resistance and treatment. BMC Medicine. 18: 1–22. https://doi.org/10.1186/s12916-020-01652-4
Ulanowska, A., Kowalkowski, T., Trawi´nska, E. & Buszewski, B. (2011). The application of statistical methods using VOCs to identify patients with lung cancer. J. Breath Res. 5: 046008. https://doi.org/10.1088/1752-7155/5/4/046008
Wang, Y., Chen, Y., Chen, X., Liang, Y., Yang, D., Dong, J., ... & Liang, Z. (2019). Angelicin inhibits the malignant behaviours of human cervical cancer potentially via inhibiting autophagy. Experimental and Therapeutic Medicine, 18(5), 3365-3374. http://doi.org/10.3892/etm.201 9.7985
Wayne, P.A. (2002). National Committee for Clinical Laboratory Standards. Performance Standards for antimicrobial susceptibility testing. 12: 01-53.
Wayne, P.A. (2018). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Clinical and Laboratory Standards Institute. 11th edition. CLSI standard M07
World Health Organization. (2017). Critically Important Antimicrobials for Human Medicine – 5th rev. Geneva: World Health Organization.
World Health Organization. (2018). Salmonella (non-typhoidal). Available at: https://www.who.int/news-room/fact-sheets/detail/Salmonella-(nontyphoidal), accessed August 27, 2020.
Yadav, V., Krishnan, A. & Vohora, D. (2020). A systematic review on Piper longum L.: Bridging traditional knowledge and pharmacological evidence for future translational research. J. Ethnophar macol., 247, 112255. https://doi.org/1 0.1016/j.jep.2019.112255
Zaveri, M., Khandhar, A., Patel, S.G. & Patel, A. (2010). Chemistry and pharmacology of Piper longum L. Int J Pharm Sci Rev Res. 5(1), 67–76.
Zhong, Z., Wheeler, M. D., Li, X., Froh, M., Schemmer, P., Yin, M., Bunzendaul, H., Bradford, B. & Lemasters, J. J. (2003). L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. Curr Opin Clin Nutr Metab Care. 6(2), 229–240. https://doi.org/10.109 7/00075197-200303000-00013
Research Articles

How to Cite

Antibacterial, antioxidant, and phytochemical analysis of Piper longum fruit extracts against multi-drug resistant non-typhoidal Salmonella strains in vitro. (2022). Journal of Applied and Natural Science, 14(4), 1225-1239. https://doi.org/10.31018/jans.v14i4.3774