In Vitro Inhibitory Potential of Lawsonia inermis Extracts against Multidrug Resistant Clinically-Relevant Bacteria: a Phytochemical, Quantitative Antimicrobial and Toxicological Assessment

Objective: Majority of the current antibiotics have become less effective due to widespread of multidrug-resistant microorganisms. Medicinal plants are promising candidates that could be used to manage this menace. Therefore, phytochemical, toxicological and antimicrobial potentiality of Lawsonia inermis extracts against MDR clinical bacteria were carried out. Material-Method: Henna leaf and seed were extracted by cold maceration technique using methanol and water and screened phytochemically. Eight MDR isolates, four of which are ESβL-producers were used for this study. In vitro antimicrobial efficacy and quantitative antimicrobial potency of extracts were estimated. MIC and MBC were determined using broth macrodilution technique. Cytotoxicity test was conducted using brine shrimp lethality assay and LC50 was determined. Results: The findings of this study revealed that aqueous leaf extract possesses maximum percentage yield of 25.58%. Tannins and phenolic compounds were detected in all extracts, while steroid was absent. Methanol seed extract showed the highest antimicrobial efficacy against all bacteria with 100 percent activity. The highest and lowest zones of inhibition were recorded at 30.0±0.00 and 10.0±0.00 mm, respectively. The zones of inhibition of extracts differed significantly. All extracts displayed highest activity index against the ESβL-producing Enterobacter aerogenes 196 that was isolated from wound with highest value at 4.28. Pseudomonas aeruginosa U109 showed maximum susceptibility index (93.75%); majority of MIC values recorded were within the range of 1.95-62.5 mg/mL. Cytotoxicity test of methanol and aqueous extracts displayed 10001000, respectively. Conclusion: Findings from this study elucidate the efficacy of Lawsonia inermis as a potential remedy to manage MDR-related infectious bacteria.

Keywords:

Antimicrobial Resistance, Multidrug Resistant Microorganisms, ESBL, Antimicrobial Activity Indigenous Plant, Phytotherapy,

PDF

___

Martin JB. The Missing Microbes: How the Overuse of antibiotics is fueling our modern plagues. Chapter five: The Wonder Drugs. HarperCollins e-books; 2014:49-60
Yang X, Ye W, Qi Y, Ying Y, Zhongni X. Overcoming Multidrug Resistance in Bacteria through Antibiotics Delivery in Surface-engineered Nano-Cargos: Recent Developments for Future Naano-Antibiotics. Front Bioeng Biotechnol. 2021;9:696514.
Esme B, Nick G, Max L, Pablo ARM, Anjela T, Diego AVP. The inequality virus: Bringing together a world torn apart by coronavirus through a fair, just and sustainable economy. Oxfam International; 2021:82 DOI: 10.21201/2021.6409.
O’Neill J. Tackling drug-resistant infections globally: final report and recommendations–The Review on Antimicrobial Resistance. Wellcome Trust; HM Government: London; 2016
Murray CJL, Ikuta KS, Sharara F, Swetschinski L, Aguiler GR, Gray A, et al. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancent. 2022.
Overbye KM, Barrett F. Antibiotics: where did we go wrong? Drug Discov Today. 2005;10(1):45-52.
WHO. Antimicrobial resistance. World Health Organization; 2021. Accessed 22/11/2021 https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance
Heinrich M, Joanne-Barnes J, Gibbons S, Kinghorn AD. Fundamentals of Pharmacognosy and Phytotherapy. Elsevier, Churchill-LivingStone; 2012:166-173
Furner-Pardoe J, Anonye BO, Cain R, Moat J, Ortori CA, Lee C, et al. Anti-biofilm efficacy of a medieval treatment for bacterial infection requires the combination of multiple ingredients. Scientific reports, natureresearch. 2020;10:12678.
Harrison F, Roberts AEL, Gabrilska R, Rumbaugh KP, Lee C, Diggle SP. A 1,000-year-old antimicrobial remedy with antistaphylococcal activity. mBio. 2015;6(4):e01129-15 doi:10.1128/mBio.01129-15
Osungunna MO. Screening of Medicinal Plants for Antimicrobial Activity: Pharmacognosy and Microbiological Perspectives. J Microbiol, Biotech & Food Sci. 2020;9(4):727-735.
El-Ghani MMA. Traditional medicinal plants of Nigeria: an overview. ABJNA 2016;7(5)220-247.
Singh YV, Regar PL, Rao SS, Jangid BL, Chand K. Potential of Planting Configuration and Water Harvesting in Improving the Production of Henna in Arid Fringes. Henna, Cultivation, Improvement and Trade. India; 2005:28-30.
Ruchi BS, Deepak KS, Sandra C, Catherine C, Alvaro V. Lawsonia inermis L. (henna): Ethnobotanical, phytochemical and phatmacological aspects. J Ethnopharmacol. 2014:1-24
Wang S, Tao Z, Li P. Lawsone suppresses azoxymethane mediated colon cancer in rats and reduces proliferation of DLD-1 cells via NF-kB pathway. Biomed Pharmacother. 2017;89:152-161
Nawagish M, Ansari SH, Ahmad S. Preliminary pharmacognostical standardization of Lawsonia inermis Linn. seeds. Res J Bot. 2007;2:161-164.
Wadankar GD, Malode SN, Sarambekar SL. Traditionally used medicinal plants for wound healing in the Washim district, Maharashtra (India). Int J PharmTech Res. 2011;3:2080–2084
Mina B, Jeevani VC, Revathy S, Pramod C, Ragav R, Manjula SN, Mruthunjaya K. Phytochemical and microscopical investigations on Lawsonia inermis roots. Int J Curr Pharm Rev Res. 2012;3:54–59
Mastanaiah J, Prabhavathi N, Varaprasad B. In vitro anti-bacterial activity of leaf extracts of Lawsonia inermis. Int J of PharmTech Res. 2011;3:1045-9
Gull I, Sohail M, Aslam SM, Athar AM. Phytochemical, Toxicological and Antimicrobial Evaluation of Lawsonia inermis Extracts against Clinical Isolates of Pathogenic Bacteria. Ann Clin Microbiol & Antimicrob. 2013;12(36):1-6
Elgailany HAM, Elnil YFH. Antibacterial Activity of Lawsonia inermis (Sudanese Henna) Leaves Extracts against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa among Recurrent Urinary Tract Infection patients in Omdurman Military Hospital. World J Pharm Sci. 2016;4(5):183-194
Muhammad HS, Muhammad S. The use of Lawsonia inermis L. (henna) in the management of burn wound infections. Afr J Biotechnol. 2005;4:934-937
Kumari P, Joshi GC, Tewani LM. Diversity and status of ethno-medicinal plants of Almora district in Uttarakhand, india. Int J Biodivers Conserv. 2011;3:298-326
Habbal O, Hasson SS, El-Hag AH, Al-Mahrooqi Z, Al-Hashim N, Al-Bimani Z, Al-Balushi MS, Al-Jabri AA. Antibacterial activity of Lawsonia inermis Linn (Henna) against Pseudomonas aeruginosa. Asian Pac J Trop Biomed. 2011:173-176
Idowu OA, Soniran OT, Ajana O, Aworinde DO. Ethnobotanical survey of antimalarial plants used in Ogun State, Southwest Nigeria. Afr J Pharm Pharmacol. 2010;4:055-060
Ojewunmi OO, Oshodi T, Ogundele OI, Micah C, Adeneke S. In vitro Antioxidant, Antihyperglycaemic and Antihyperlipidaemic Activities of Ethanol Extract of Lawsonia inermis Leaves. British J Pharm Res. 2014;4(3):301-314.
Tiwari P, Kumar B, Kaur M, Kaur G, Kaur H. Phytochemical Screening and Extraction: A Review. Int Pharm Sci. 2011;1(1):98-106
Geetha DH, Indhiramuthu J, Rajeswari M. Micro-morphological and phytochemical studies of aerial parts of Indigofera enneaphylla Linn. J Pharmacogn Phytochem. 2016;5(1): 216-220
Singh V, Kumar R. Study of Phytochemical Analysis and Antioxidant Activity of Allium sativum of Bundelkhand Region. Int J Life-sci Sci Res. 2017;3(6):1451-1458.
Sofowora A, Ogunbodede E, Onayade A. The role and place of medicinal plants in the strategies for disease prevention. Afr J Trad, Complement & Altern Med. 2013;10(5):210-229.
Pandey A, Tripathi S. Concepts of Standardization, Extraction and Prephytochemical Screening Strategies for Herbal drug. J Pharmacog Phytochem. 2014;2(5):115-119
Silva GO, Abeysundara AT, Aponso M. Extraction methods, Quantitative and Qualitative Techniques for Screening of Phytochemicals from plants. American J Essent Oil. 2017;5(2):29-32
Ali KS, Al-hood FA Obad K, Alshakka M. Phytochemical Screening and Antibacterial (Lawsonia inermis) against some Bacterial Pathogens. IOSR J Pharm Biol Sci. 2016;11(2)III:24-27.
Rajput M, Kumar N. In vitro Antimicrobial and Antibiofilm Efficacy of Medicinal Plant Extracts against Clinical MDR Isolates from Scalp Infection Cases. Int J Sci Technol Res. 2020;9(2):4216-4228
Idowu PA. Antibacterial Activity of Crude Extracts and Alkaloidal Fractions of Argemone meicana Linn. (Papaveraceae). Nigerian J Sci. 2012;46:23-28
Andrews JM. Determination of minimum inhibitory concentration. J Antimicrob Chemother. 2001;48(1):5-16.
Cheesbrough M. District laboratory practice in tropical countries. Part 2nd Ed. Cambridge University Press, New York; 2006:137-138.
McMurray RL, Ball MEE, Tunney MM, Corcionivoschi N, Situ C. Antibacterial activity of four plants extracts extracted from Traditional Chinese Medicinal plants against Listeria monocytogenes, Escherichia coli, and Salmonella enterica subsp. enterica serovar enteritidis. Microorganisms. 2020;8(962).
Rani P, Khullar N. Antimicrobial Evaluation of some Medicinal Plants for their Anti-enteric Potential aginst Multi-drug Resistant Salmonella typhi. Phytother Res. 2004;18:670-673.
Al-Rubiay KK, Jaber NN, Al-Mhaawe BH, Alrubaiy LK. Antimicrobial Efficacy of Henna Extracts. Oman Med J. 2008;23(4):253-256
Kannahi M, Vinotha K. Antimicrobial activity of Lawsonia inermis leaf extracts against some human pathogens. Int J Curr Microbiol Appl Sci. 2013;2(5):342-349
Shahabinejad S, Kariminik A. Antibacterial activity of methanol extract of Lawsonia inermis against uropathogenic bacteria. MicroMedicine. 2019;7(2):31-36
Usman RA, Rabiu U. Antimicrobial Activity of Lawsonia inermis (Henna) Extracts. Bayero J Pure Appl Sci: Special Conference Edition, November. 2018;11(1):167-171
Harrison F, Furner-Pardoe J, Connelly E. An assessment of the evidence for antibacterial activity of stinging nettle (Urtica dioica) extracts. Access Microbiology. 2022;4:000336 DOI 10.1099/acmi.0.000336
Khameneh B, Iranshahy M, Soheili V, Bazzaz BSF. Review on plant antimicrobials: a mechanistic viewpoint. Antimicrob Resistance Infection Control. 2019;8(118):1-28.
Maitera ON, Louis H, Oyebanji OO, Anumah AO. Investigation of tannin content in Diospyros mespiliformis extract using various extraction solvents. J Anal Pharm Res. 2018;7:1.
El-Mahmood MA. Antibacterial activity of crude extracts of Euphorbia hirta against some bacteria associated with enteric infections. J Med Plants Res. 2009;3(7):498-505
Nguyi AA. Tannins of some Nigerian flora. Nigerian J Biotechnol. 1988;6:221-226
Nature’s Pharmacopeia. A World of Medicinal Plants. Dan Choffnes (eds). Columbia University Press. New York; 2016:58-81;317,321,327
Ibrahim SMS, Rasool CS, Al-Asady AA. Antimicrobial activity of crude extract against Gram-positive bacteria. Iraq Med J. 2021;5(3):89-93.
Soliman SSM, Alsaadi AI, Youssef EG, Khitrov G, Noreddin AM, Husseiny MI, Ibrahim AS. Calli essential oils synergize with lawsone against multidrug resistant pathogens. Molecules. 2017:22(2223):1-13.
Agunloye OM, Oboh G. Effect of different processing methods on antihypertensive property and antioxidant activity of sandpaper leaf (Ficus exasperata) extracts. J Diet Suppl. 2018;15(6):871-883.
Sharmeen R, Hossain N, Rahman M, Foysal J, Miah F. In-vitro antibacterial activity of herbal aqueous extract against multi-drug resistant Klebsiella sp. Isolated from human clinical samples. Int Curr Pharm J. 2012:1(6):133-137
Aqil F, Ahmad I. Antibacterial properties of traditionally used Indian medicinal plants. Methods Find Exp Clin Pharmacol. 2007;29(2):79-92.
Eloff JN. Avoiding pitfalls in determining antimicrobial activity of plant extracts and publishing the results. BMC Complement Altern Med. 2019;19(106):1-8.
Daemi A, Farahpour MR, Oryan A, Karimzadeh S, Tajer E. Topical administration of hydroethanolic extract of Lawsonia inermis (henna) accelerates excisional wound healing process by reducing tissue inflammation and amplifying glucose uptake. Kaohsiung J Med Sci. 2019;35:24-32.
Thormar H. Lipids and essential oils as antimicrobial agents. John Wiley & Sons Ltd, The Atrim, South Gate, Chichester, West Sussex, United Kingdom; 2011:212-213.
Al-kurashy HMK, Ai-windy SA, Al-buhadilly AK. Evaluation of the antimicrobial activity of Lawsonia inermis: in vitro study. Iraqi J Sci. 2011;52:16-19.
Michael AS, Thompson CG, Abramovittz M. Artemia salina as a Test Organism for Bioassay. Science. 1956;123(3194):464.
Meyer BN, Ferrigni NR, Putnam JE, Jacobsen LB, Nichols DE, McLaughlin JL. Brine Shrimp: A convenient general bioassay for active plant constituents. Planta Medica. 2011;45:31-34.
Waghulde S, Kale MK, Patil VR. Brine Shrimp Lethality Assay of the Aqueous and Ethanolic Extracts of the Selected Species of Medicinal Plants. MDPI proceedings. 2019;41: 47.
Nerdy N, Lestari P, Sinaga JP, Ginting S, Zebua NF, Mierza V, Bakri TK. Brine Shrimp (Artemia salina Leach.) Lethality Test of Ethanolic Extract from Green Betel (Piper betle Linn.) and Red Betel (Piper crocatum Ruiz and Pav.) through the Soxhletation Method for Cytotoxicity Test. Journal of Medical Sciences 2021;9(A):407-412.
Clarkson C, Maharaj VJ, Crouch NR, Grace OM, Pillay P, Matsabisa MG, et al. In vitro antiplasmodial activity of medicinal plants native to or naturalized in South Africa. J Ethnopharm. 2004;92:177-191.
Gosselin RE, Smith RP, Hodge HC, Braddock J. Clinical Toxicology of Commercial Products. Williams & Wilkins, Balyimore; 1984:5.
Karchesy YM, Kelsey RG, Constantine G, Karchesy JJ. Biological screening of selected Pacific Northwest forest plants using the brine shrimp (Artemia salina) toxicity bioassay. Springer Plus. 2016;5(510):1-9.