Zanco Journal of Medical Sciences
Skip Navigation Links
Available Issues
Editorial Board
Information for Authors
Review Process
Links and Contacts
  Zanko J Med Sci:  Apr. 2017; 21 (1): 1664-1672

Antibacterial and antibiofilm activity of Ammi majus seed against Gram-positive bacteria

Aveen Nozad Adham & Zheen Abdulelah Abdulah


Background and objective: Ammi majus belongs to family Apiaceae is an important plant used in a different country traditionally for treatment bacterial infection. This study aimed to investigate in vitro antibacterial and antibiofilm activity of Ammi majus seed extracts against six isolated Gram-positive bacteria.

Methods: The antibacterial activity of seed extracts were screened by disk diffusion and agar overlay bioautography method and their activities were further determined by minimum inhibitory and minimum bactericidal concentration. Biofilm formation was evaluated by the microtiter plate crystal violet assay. The quantity of bound bacteria was determined by measuring absorbance at (OD630 nm) using microtiter plate reader.

Results: Ethanol 80% and ethyl acetate extracts showed variable activity against all isolated bacteria while petroleum ether extract revealed resistance against studied bacteria. Streptococcus mitis showed more pronounced sensitivity against both extracts by disk diffusion (12 mm and 10 mm) and bioautography method with minimum inhibitory concentration 7.8125 mg/ml of ethanol 80% and 15.625 mg/ml for ethyl acetate extracts. Ethanol 80% and ethyl acetate extract significantly reduced biofilm formation as compared to control, showed antibiofilm activity at 0.4883-62.5 mg/ml and exhibited strongest antibiofim activity against Staphylococcus aureus.

Conclusion: Ammi majus seed extracts revealed highest antibacterial activity against Streptococcus species and strongest antibiofilm activity on Staphylococcus species.

Keywords: Ammi majus; Disk diffusion; Bioautography; minimum inhibitory concentration; Antibiofilm.


1. Guarrera PM. Traditional phytotherapy in Central Italy (Marche, Abruzzo, and Latium). Fitoterapia 2005; 76:1–25.

2. Høiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O. Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 2010; 35(4):322–32.

3. Hugo WB, Russell AD. Pharmaceutical Microbiology, 7th edn. USA: Blackwell publishing company; 2004. p. 3–38.

4. Parsek M, Greenberg. Socio microbiology: the connections between quorum sensing and biofilms. Trends in Microbiol 2004; 13:3–27.

5. Roland PS. Chronic suppurative otitis media: a clinical overview. Ear Nose Throat J 2002; 81:8–10.

6. Head NE, Yu HW. Cross-sectional analysis of clinical and environmental isolates of Pseudomonas aeruginosa: Biofilm formation, virulence and genome diversity. Infect Immun 2004; 72:133–44.

7. Essawi T, Srour M. Screening of some Palestinian medicinal plants for antibacterial activity. J Ethnopharmacol 2000; 70:343–9.

8. Knetsch MLW, Koole LH. New strategies in the development of antimicrobial coatings: the example of increasing usage of silver and silver nanoparticles. Polym 2011; 3:340–66.

9. Sagar S, Kaistha DS. Virulence profile of an emerging coagulase negative Staphylococcus auricularis NC clinical isolate. J Pure Appl Microbiol 2011; 5(2):787–92.

10. Nostro A, Roccaro AS, Bisignano G, Marino A, Cannatelli MA, Pizzimenti FC et al. Effects of oregano, carvacrol, and thymol on Staphylococcus aureus and Staphylococcus epidermidis biofilms. J Med Microbiol 2007; 56(4):519–23.

11. Standar K, Kreikemeyer B, Redanz S, Münter LW, Laue M,  Podbielski A. Setup of an in vitro test system for basic studies on biofilm behavior of mixed-species cultures with dental and periodontal pathogens. PLOS One 2010; 5(10):1–14.

12. Central Council for Research in Unani Medicine. Standardization of single drugs of Unani medicine – Part I. New Delhi, Ministry of Health and Family Welfare; 1987. p. 182–4.

13. Saeed MA, Khan F Z. Studies on the contact dermatitis properties of Indigenous Pakistani   medicinal Plants. J Faculty Pharm Gazi Univer 1994; 11(1):17–24.

14. Lin J, Zhan SM, Wu K, Willett WC, Fuchs CS, Giovannucci E. Flavonoid intake and colorectal cancer in men and women. Am J Epidemiol 2006; 164:644–51.

15. Abdulla WA. Preliminary studies on the anti-schistosomal effect of Ammi majus L. Egypt J Bilharziasis 1978; 4:19–26.

16. Shihab H. Mutlag. Dose-Dependent Anti-inflammatory effect of Ammi majus alcoholic extract in the rat: Chronic study. Iraqi J Pharm Sci 2012; 21(1):82–6.

17. Alupuli A, Calinescu I, Lavric V. Ultrasonic vs. microwave extraction intensification of active principles from medicinal plants. AIDIC Conference Series 2009; 09:1–8.

18. Maidment C, Dyson A, Haysom I. A study into the antimicrobial effects of cloves (Syzygium aromaticum) and cinnamon (Cinnamomum zeylanicum) using disc diffusion assay. Nutr Food Sci 2006; 36(4):225–30.

19. Silva MT, Simas SM, Batista TG, Cardarelli P, Tomassini TC. Studies on antimicrobial activity, in vitro, of Physalis angulata L.(Solanaceae) fraction and physalin B bringing out the importance of assay determination. Mem Inst Oswaldo Cruz 2005; 100:779–82.

20. Teanpaisan R, Senapong S, Puripattanavong J. In vitro antimicrobial and antibiofilm activity of Artocarpus Lakoocha (Moraceae) Extract against Some Oral Pathogens. Trop J Pharm Res 2014; 13 (7):1149–55

21. Raja A, Ali F, Khan I. Antistaphylococcal and biofilm inhibitory activities of acetyl-11-keto-β-boswellic acid from Boswellia serrata. BMC Microbiol 2011; 11:54–62.

22. Dogruoz N, Zeybek Z, Karogoz A. Antibacterial activity of some plants extracts. J Biol 2008; 67(1):17–21.

23. Al-Hadidi AK, Al-Numan YA, Al-Daody CA. The interaction between some phenolic compounds in Ammi majus herb: (khillah) extracts and antibiotics against some selected bacterial isolates in vitro. Raf J Sci 2013; 24(2):17–30.

24. Bavington C, Page C. Stopping bacterial adhesion: a novel approach to treating infections. Respir 2005; 72:335–44.

25. Dean SN, Bishop BM, van Hoek ML. Natural and synthetic cathelicidin peptides with anti-microbial and anti-biofilm activity against Staphylococcus aureus. BMC Microbiol 2011; 11:114.

26. Ergün B, Çoban T, Onurdag F, Banoglu E. Synthesis, antioxidant and antimicrobial evaluation of simple aromatic esters of ferulic acid. Arch Pharmacal Res 2011; 34(8):1251–61.

27. Nostro A, Marino A, Blanco AR. In vitro activity of carvacrol against Staphylococcal preformed biofilm by liquid and vapor contact. J Med Microbiol 2009; 58(6):791–7.

28. Lee JH, Park JH, Cho HS, Joo SW, Cho MH, Lee J. Anti-biofilm activities of quercetin and tannic acid against Staphylococcus aureus. Biofouling 2013; 29(5):491–9.

29. Adham NA. Comparative extraction methods, fluorescence, qualitative and quantitative evaluation of Ammi majus seed extracts. J Pharmacogn Phytochem 2015; 4(1):41–4.

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.