Bactericidal Versus Bacteriostatic Activity of Origanum vulgare and Andrographis paniculata Extracts Against Streptococcus spp.: An in Vitro MBC/MIC Ratio-Based Assessment

Farah Razzaq Kbyeh; Mohamed Abdul Rida Yaseen

doi:10.70604/jmtbas.v3i2.196

Authors

Farah Razzaq Kbyeh Department of Clinical Laboratory Science, College of Pharmacy, University of Al-Qadisiyah, Al-Diwaniyah, Iraq.
Mohamed Abdul Rida Yaseen Department of Pharmaceutical Chemistry, College of Pharmacy, University of Al-Qadisiyah, Al-Diwaniyah, Iraq.

Abstract

Background and Objective: There is a growing emergence of antibiotic resistance bacteria and there is a need to search for new antimicrobial agents from natural sources. The present study was designed to evaluate and compare the antibacterial activity of ethanolic leaf extracts of Origanum vulgare and Andrographis paniculata against clinical isolates of Streptococcus spp. at concentrations of 50, 100, 200 and 400 mg/mL and compare their efficacy with standard reference antibiotics (Cloxacillin 30 µg, Penicillin 6 µg, Amoxicillin 25 µg and Ciprofloxacin 5 µg).

Methods: Antibacterial activity was evaluated by agar well diffusion method and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were calculated by broth microdilution method following NCCLS standards. To determine the nature of the antimicrobial effect as bactericidal or bacteriostatic, the MBC/MIC ratio was determined.

Results: Both plant extracts showed concentration dependent antibacterial activity against Streptococcus spp. The inhibition zone of O. vulgare ethanolic extract (13.73–44.89 mm) was substantially higher than that of A. paniculata (10.18–18.72 mm) at all tested concentrations (p < 0.05). At the highest concentration (400 mg/mL) O. vulgare extract was more effective than that of Cloxacillin and Ciprofloxacin i.e. 44.89 ± 0.92 mm vs 38.12 ± 0.65 mm and 38.47 ± 0.17 mm. For O. vulgare, the MIC and MBC values were 0.156 and 0.624 mg/mL (MBC/MIC ratio = 4.0, bactericidal), respectively, while for A. paniculata, the MIC and MBC values were 0.624 and 5.0 mg/mL (MBC/MIC ratio = 8.0, bacteriostatic), respectively.

Conclusion: The ethanolic extract of O. vulgare displayed a higher bactericidal effect on Streptococcus spp. compared to A. paniculata and several conventional antibiotics, showing its potential as a natural antibacterial agent. Further phytochemical characterisation and in vivo research are justified

Keywords: ,

Keywords:

Origanum vulgare, Andrographis paniculata, Streptococcus, antibacterial activity, MIC, MBC, agar well diffusion, plant extract, antimicrobial resistance

DOI

https://doi.org/10.70604/jmtbas.v3i2.196

References

[1] Z. Xu, Z. Qiu, Q. Liu, et al., Converting organosulfur compounds to inorganic polysulfides against resistant bacterial infections, Nat. Commun. 9 (2018) 3713.

[2] R.L. Pacheco, C.D.O.C. Latorraca, A.P. Pires dos Santos, et al., Efficacy and safety of home-based intravenous antibiotic therapy among adults: a systematic review, Int. J. Antimicrob. Agents 59 (2022) 106555.

[3] G. Tiseo, G. Brigante, D.R. Giacobbe, et al., Diagnosis and management of infections caused by multidrug-resistant bacteria, Int. J. Antimicrob. Agents 60 (2022) 106611.

[4] E. Adodo, D. Ahamioje, Y.I. Ibitoye, I. Matthew, Antibiogramic efficacy of Cymbopogan citratus, Ocimum gratissimum, and Kalanchoe pinnata methanolic and aqueous extracts on colistin-resistant non-clinical isolates of gram-negative bacteria, UMU Scientifica 3 (2024) 102.

[5] S. Safarzadeh, F. Shirban, M. Bagherniya, T. Sathyapalan, A. Sahebkar, The effects of herbal medicines on cancer therapy-induced oral mucositis, Phytother. Res. 36 (2022) 243–265.

[6] A.A. Alfuraydi, I.M. Aziz, F.N. Almajhdi, Assessment of antioxidant, anticancer, and antibacterial activities of the rhizome of ginger, J. King Saud Univ.- Sci. 36 (2024) 103112.

[7] P.S.X. Yap, B.C. Yiap, H.C. Ping, S.H.E. Lim, Essential oils, a new horizon in combating bacterial antibiotic resistance, Open Microbiol. J. 8 (2014) 6–14.

[8] G. Chikowe, L. Mpala, I.E. Cock, Eucalyptus grandis leaf and flower extracts lack antibacterial activity and are non-toxic in vitro, Pharmacognosy Commun. 10 (2019) 23–29.

[9] A. Sharma, R.D.C. Flores-Vallejo, A. Cardoso-Taketa, M.L. Villarreal, Antibacterial activities of medicinal plants used in Mexican traditional medicine, J. Ethnopharmacol. 208 (2017) 264–329.

[10] M. Alekseeva, T. Zagorcheva, I. Atanassov, K. Rusanov, Origanum vulgare L.—A review on genetic diversity, cultivation, biological activities and perspectives for molecular breeding, Bulg. J. Agric. Sci. 26 (2020) 1183–1197.

[11] R. Nurzyńska-Wierdak, M. Walasek-Janusz, Chemical Composition, Biological Activity, and Potential Uses of Oregano (Origanum vulgare L.) and Oregano Essential Oil, Pharmaceuticals 18 (2025) 267.

[12] A. Chbel, A. Elmakssoudi, M. Rey-Méndez, et al., Analysis of the chemical compositions of six essential oils and evaluation of their antioxidant and antibacterial activities, J. Herbmed Pharmacol. 11 (2022) 401–408.

[13] S. Soltani, A. Shakeri, M. Iranshahi, M. Boozari, A Review of the Phytochemistry and Antimicrobial Properties of Origanum vulgare L. and Subspecies, Iran J. Pharm. Res. 20 (2021) 268–285.

[14] Z. Ilić, L. Stanojević, L. Milenković, et al., The Yield, Chemical Composition, and Antioxidant Activities of Essential Oils from Different Plant Parts of the Wild and Cultivated Oregano, Horticulturae 8 (2022) 1042.

[15] J. Mesmar, R. Abdallah, A. Badran, M. Maresca, E. Baydoun, Origanum syriacum Phytochemistry and Pharmacological Properties: A Comprehensive Review, Molecules 27 (2022) 4272.

[16] M. Alagawany, M.E. Abd El-Hack, M.R. Farag, et al., The applications of Origanum and its derivatives in human, ruminant and fish nutrition—A review, Ann. Anim. Sci. 20 (2020) 389–407.

[17] J.P. Veenstra, J.J. Johnson, Oregano (Origanum vulgare) extract for food preservation and improvement in gastrointestinal health, Int. J. Nutr. 3 (2019) 43.

[18] M. Walczak, M. Michalska-Sionkowska, D. Olkiewicz, et al., Potential of Carvacrol and Thymol in Reducing Biofilm Formation on Technical Surfaces, Molecules 26 (2021) 2723.

[19] S. Peter, N. Sotondoshe, B.A. Aderibigbe, Carvacrol and Thymol Hybrids: Potential Anticancer and Antibacterial Therapeutics, Molecules 29 (2024) 2277.

[20] K. Kachur, Z. Suntres, The antibacterial properties of phenolic isomers, carvacrol and thymol, Crit. Rev. Food Sci. Nutr. 60 (2020) 3042–3053.

[21] F.P.K. Jesus, L. Ferreiro, K.S. Bizzi, et al., In vitro activity of carvacrol and thymol combined with antifungals or antibacterials against Pythium insidiosum, J. Mycol. Médicale 25 (2015) 89–93.

[22] S. Hossain, Z. Urbi, R. Capasso, Antibiofilm Activity of Andrographis paniculata, Andrographolide and Its Derivatives: A Systematic Review, Life 11 (2021) 348.

[23] N.F. Sholihah, L.S. Saula, G.S. Mally, Comparison of Antibacterial of Sambiloto (Andrographis paniculata) Leaf and Kemangi (Ocimum sanctum) Leaf Extract Against Staphylococcus aureus, J. Pharm. Sci. 5 (2022) 279–85.

[24] M. Majumdar, T.K. Misra, D.N. Roy, In Vitro Anti-Biofilm Activity of 14-Deoxy-11,12-Didehydroandrographolide from Andrographis paniculata against Pseudomonas aeruginosa, Braz. J. Microbiol. 51 (2020) 15–27.

[25] M. Fitzgerald, M. Heinrich, A. Booker, Medicinal Plant Analysis: A Historical and Regional Discussion of Emergent Complex Techniques, Front. Pharmacol. 10 (2020) 1480.

[26] C. Rodríguez-Melcón, C. Alonso-Calleja, C. García-Fernández, et al., Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for Twelve Antimicrobials, Biology 11 (2022) 46.

[27] F.K. Razzaq, Investigation the effect of alone and combined diclofenac and ciprofloxacin against urinary tract infection induced by resistant E. coli (O157-H7) in female rabbits, Doctoral dissertation, University of Baghdad (2024).

[28] M.A. Salam, M.Y. Al-Amin, M.T. Salam, et al., Antimicrobial Resistance: A Growing Serious Threat for Global Public Health, Healthcare 11 (2023) 1946.

[29] L. Tao, Y. Liang, Z. Xia, et al., Antibacterial activities of oregano essential oils and their active components, Front. Pharmacol. 16 (2025) 1579283.

[30] C.-M. Radu, C.C. Radu, I.-D. Venter, et al., Antimicrobial Activity of Oregano, Thyme, and Lavender Oils Against Oral Pathogens, Oral 5 (2025) 87.

[31] J.A. Avbunudiogba, S.E. Okafo, C.L. Nwobi, Antimicrobial investigation, Formulation and Evaluation of Andrographis paniculata aqueous herbal cream for topical application, Res. J. Pharm. Tech. 15 (2022) 3553–8.

[32] D. Gwiazdowska, A. Waśkiewicz, K. Juś, et al., Antimicrobial and Antibiofilm Activity of Origanum vulgare Extracts Obtained by Supercritical Fluid Extraction, Molecules 29 (2024) 5823.

[33] S. Tejada-Muñoz, D. Cortez, J. Rascón, et al., Antimicrobial Activity of Origanum vulgare Essential Oil against Staphylococcus aureus and Escherichia coli, Pharmaceuticals 17 (2024) 1430.

[34] S. Hossain, Z. Urbi, H. Karuniawati, et al., Andrographis paniculata: An Updated Review of Phytochemistry, Antimicrobial Pharmacology, and Clinical Safety, Life 11 (2021) 348.

[35] T.M. Uddin, A.J. Chakraborty, A. Khusro, et al., Antibiotic resistance in microbes: History, mechanisms, therapeutic strategies and future prospects, J. Infect. Public Health 14 (2021) 1750–1766.

[36] S.K. Sah, U. Rasool, S. Hemalatha, Andrographis paniculata extract inhibit growth, biofilm formation in multidrug resistant strains of Klebsiella pneumoniae, J. Tradit. Complement. Med. 10 (2019) 599–604.

[37] A. Rakhmawati, K.R. Pertiwi, Phytochemical Analysis of Andrographis paniculata Extract and Its Potential as Anti Bacterial Growth and Anti Biofilm Formation, J. Microbiol. Biotechnol. Food Sci. 15 (2025) e11968.

[38] M. Chroho, Y. Rouphael, S.A. Petropoulos, L. Bouissane, Carvacrol and Thymol Content Affects the Antioxidant and Antibacterial Activity of Origanum compactum and Thymus zygis Essential Oils, Antibiotics 13 (2024) 139.

[39] M.A. Mudasir, A. Saleem, J. Nazir, et al., Synthesis and pharmacological evaluation of Andrographolide and Ajwain as promising alternatives to antibiotics, Int. Immunopharmacol. 142 (2024) 113163.

[40] Z. Ren, Z. Chen, Y. Xie, P. Coghi, Andrographolide and Its Derivatives: A Comprehensive Review of Anti-Infective Properties and Clinical Potential, Molecules 30 (2025) 4273.

[41] S.D. Cox, C.M. Mann, J.L. Markham, et al., The mode of antimicrobial action of the essential oil of Melaleuca alternifolia (tea tree oil), J. Appl. Microbiol. 88 (2000) 170–175.

[42] A. Ultee, M.H.J. Bennik, R. Moezelaar, The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus, Appl. Environ. Microbiol. 68 (2002) 1561–1568.