Antimicrobial Activity of Combined Extracts of Trachyspermum, Thymus and Pistachio against Some Pathogenic Bacteria

Document Type : Original Article


1 MSc of Microbiology,Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran

2 Associate Professor of Microbiology, Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran


Background: Microbial biofilms are responsible for many human infections and increase of antibiotic resistant bacteria. Therefore, finding an efficient way to prevent infection and biofilm formation of bacteria is essential. Medicinal plants are among suitable candidates to inhibit biofilm formation of bacteria. The aim of this study was to evaluate the antimicrobial effects of Trachyspermum, Thymus and Pistachio (T.T.P) combined extracts (methanolic and ethanolic) against six pathogenic bacteria in planktonic and biofilm forms.
Methods: The antibacterial activities of T.T.P combined extracts against planktonic form of pathogenic bacteria were evaluated by disc diffusion and macro-broth dilution methods. Also, anti-biofilm activity of the combined extracts was evaluated by microtiter plate method.
Results: The results of disc diffusion analysis (MBC and MIC) showed that both methanolic and ethanolic extracts of T.T.P combined extracts were efficient for inhibition of planktonic forms of bacteria, although ethanol extract was more effective compared to the methanolic extract. The T.T.P combined extracts had inhibitory effect against biofilm formation of pathogenic bacteria. Also, these extracts efficiently demolished biofilm formation and prevent metabolic activity of bacteria in biofilm structures. The concentration of each extract had a direct relationship with its inhibitory effect. The maximum inhibitory effects of T.T.P combined extracts on biofilm formation, demolish of biofilm structure and inhibition of the metabolic activity of bacteria in the biofilm were 95.3% for P. aeruginosa, 97.2% for P. aeruginosa and 81.16% for E. coli.
Conclusion: The results of this research demonstrated that T.T.P combined extracts can be used as an alternative component with inhibitory effect against pathogenic bacteria in planktonic and biofilm forms.


  1. Mohsenipour Z, Hassanshahian M. Antibacterial Activity of Euphorbia hebecarpa Alcoholic Extracts against Six Human Pathogenic Bacteria in Planktonic and Biofilm Forms, Jundi J Microbiol; 2016; 9(6):e34701.
  2. Izquierdo-Cubas F, Zambrano A, Frometa I, Gutierrez A, Bastanzuri M, Guanche H, et al. National prevalence of nosocomial infections. Cuba 2004. J Hosp Infect 2008; 68(3):234-40.
  3. Sadeghian I, Hassanshahian M, Sadeghian S, Jamali S. Antimicrobial Effects of Quercus Brantii Fruits on Bacterial Pathogens, Jundi J Microbiol 2012; 5(3): 465-469.
  4. Sepehri Z, Hassanshahian M, Shahi Z, Nasiri A, Baigi S. Antibacterial Effect of Ethanol Extract of Camellia Sinensis L against Escherichia Coli, Asi Pacific J Microbiol Res 2014; 2 (1): 6-8.
  5. Hassanshahian M, Bayat Z, Saeidi S, Shiri Y. Antimicrobial activity of Trachyspermum ammi essential oil against human bacterial, Inter J Advan Biol Biomed Res 2014; 2(1); 18-24.
  6. Szczepanski S, Lipski A. Essential oils show specific inhibiting effects on bacterial biofilm formation. Food Control 2014; 36(1):224-9.
  7. Saeidi S, Shiri Y, Bokaeian M, Hassanshahian M. Antibacterial Activity of Essential Oil of Saturejahortensis Against Multi-Drug Resistant Bacteria, Inter J Enteric Path 2013; 2(2): 1-4.
  8. Fricks-Lima J, Hendrickson CM, Allgaier M, Zhuo H, Wiener-Kronish JP, Lynch SV, et al. Differences in biofilm formation and antimicrobial resistance of Pseudomonas aeruginosa isolated from airways of mechanically ventilated patients and cystic fibrosis patients. Int J Antimicrob Agents 2011; 37(4):309-15.
  9. Bokaeian M, Sheikh M, Hassanshahian M, Saeidi S, Sahraei S. The antibacterial activity of silver nanoparticles produced in the plant sesamum indicum seed extract: a green method against multi-drug resistant Escherichia coli, Inter J Enteric Path 2014; 2(2): e17928.
  10. Savithramma N, Rao ML, Suhrulatha D. Screening of medicinal plants for secondary metabolites. Middle-East Journal of Scientific Research 2011; 8(3):579-84.
  11. Rezaie Keikhaie K, Ghorbani S, Hosseinzadeh Z, Hassanshahian M. Antimicrobial activity of methanol extract of Citrullus colocynthis against antibiotic-resistant Staphylococcus aureus, Advan Herb Med 2017; 3 (3): 1-6.
  12. Chakraborty B, Nath A, Saikia H, Sengupta M. Bactericidal activity of selected medicinal plants against multidrug resistant bacterial strains from clinical isolates. Asian Pac J Trop Med 2014; 7S1:S435-41.
  13. Saeidi S, Amini Boroujeni N, Ahmadi H, Hassanshahian M. Antibacterial Activity of Some Plant Extracts Against Extended- Spectrum Beta-Lactamase Producing Escherichia coli Isolates, Jundishapur J Microbiol. 2015; 8(2):e15434.
  14. Weinstein RA. Controlling antimicrobial resistance in hospitals: infection control and use of antibiotics. Emerg Infect Dis 2001; 7(2):188-92.
  15. Mohsenipour Z, Hassanshahian M. The inhibitory effect of Thymus vulgaris extracts on the planktonic form and biofilm structures of six human pathogenic bacteria, Avicen J Phytomed 2015; 5 (4): 309-317.
  16. Fayad NK, AL-Obaidi OH, Al-Noor TH, Ezzat MO. Water and alcohol extraction of Thyme plant (Thymus vulgaris) and activity study against bacteria, tumors and used as anti-oxidant in margarine manufacture. Innovative Systems Design and Engineering 2013; 4(1):41-51.
  17. Hassanshahian M, Khosravi F. Study the antimicrobial effects of artemisia santonica extract on some pathogenic bacteria, Advan Herb Med 2015; 1(4): 43-46.
  18. Rezaie Keikhaie K, Bagheri G, Hassanshahian M, Saeidi S. Antimicrobial effects of zataria multiflora essential oils on acinetobacter strains isolated from clinical specimens, J Herb Drugs 2018; 8 (4): 251-256.
  19. Mohsenipour Z, Hassanshahian M. Antibacterial activity of Espand (Peganum harmala) alcoholic extracts against six pathogenic bacteria in planktonic and biofilm forms. Biol J Microorgan 2016; 4(16): 47-57.
  20. Zarshenas MM, Moein M, Samani SM, Petramfar P. An overview on ajwain (Trachyspermum ammi) pharmacological effects; modern and traditional. Journal of Natural Remedies 2013; 14(1):98-105.
  21. Javadian F, Sepehri Z, Saeedi S, Hassanshahian M. Antifungal effects of the extract of the Withania somnifera on Candida albicans, Adv Herb Med 2016; 2(1): 32-43.
  22. Mohsenipour Z, Hassanshahian M, Moradi M. Investigations of Antimicrobial Activity of Eucalyptus Camaldulensis Extracts against Six Pathogenic Bacteria in Planktonic Form and Biofilm. J Kerman Uni Med Scien 2015; 22 (2): 172-184.
  23. Sepehri Z, Javadian F, Khammari D, Hassanshahian M. Antifungal effects of the aqueous and ethanolic leaf extracts of Echinophora platyloba and Rosmarinus officinalis, Curr Med Mycol 2016; 2(1): 16-25..
  24. Yoshida H, Katsuzaki H, Ohta R, Ishikawa K, Fukuda H, Fujino T, et al. Antimicrobial activity of the thiosulfinates isolated from oil-macerated garlic extract. Biosci Biotechnol Biochem 1999; 63(3):591-4.
  25. Hamayeli H, Shoshtari A, Hassanshahian M, Askari M. Study the antimicrobial activity of six marine sponges and three parts of sea anemone on Candida Albicans, J Coas Life Med 2016; 4(8): 122-129.
  26. Jahani Z, Hosseinzadeh F, Shahi Z, Shikhzadeh M, Hassanshahian M, Saeedi S. In vitro study of antimicrobial effects of Rosmarinus officinalis and Glycyrrhiza glabra extracts against some pathogens, Adv Herb Med 2016; 2(4): 32-39.
  27. Mashhadi M, Fakhri J, Saeedi S, Hassanshahian M, Abkhoo A. Antimicrobial Effects of Medicinal Plants Collected in Zabol, Iran, on Pathogenic Food Pathogenic, J Med Bacteriol 2016; 5(3): 18-28.
  28. Masoumipour F, Hassanshahian M. Antimicrobial Activity of Five Medicinal Plants on Candida albicans, Iran J Toxicol 2016; 10(6): 65-77.
  29. Ramage G, Lopez-Ribot JL. Techniques for antifungal susceptibility testing of Candida albicans biofilms. Methods Mol Med 2005; 118:71-9.
  30. Mohsenipour Z, Hassanshahian M. Investigating the Effectiveness of Centaurea cyanus Extracts on Planktonic Growth and Biofilm Structures of Six Pathogenic Bacteria, SSU J 2014; 22 (4): 1358-1370.
  31. Trentin Dda S, Giordani RB, Zimmer KR, da Silva AG, da Silva MV, Correia MT, et al. Potential of medicinal plants from the Brazilian semi-arid region (Caatinga) against Staphylococcus epidermidis planktonic and biofilm lifestyles. J Ethnopharmacol 2011; 137(1):327-35.
  32. Heydari F, Saeedi S, Hassanshahian M. Antibacterial activity of Mentha longifolia against Salmonella typhimurium, Adv Herb Med 2015; 1 (3): 42-47.
  33. Ghotaslou R, Salahi B. Effects of oxygen on in-vitro biofilm formation and antimicrobial resistance of pseudomonas aeruginosae. Pharm Sci 2013; 19(3):96-9.
  34. Nascimento GG, Locatelli J, Freitas PC, Silva GL. Antibacterial activity of plant extracts and phytochemicals on antibiotic-resistant bacteria. Braz J Microbiol 2000; 31(4):247-56.
  35. Sienkiewicz M, Lysakowska M, Denys P, Kowalczyk E. The antimicrobial activity of thyme essential oil against multidrug resistant clinical bacterial strains. Microb Drug Resist 2012; 18(2):137-48.
  36. Tariq M, Gore M, Aruna K. Antibacterial and synergistic activity of ethanolic ajwain (trachyspermum ammi) extract on ESBL and MBL producing uropathogens. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6(6):278-84.
  37. Tohidi M, Khayami M, Nejati V, Meftahizade H. Evaluation of antibacterial activity and wound healing of Pistacia atlantica and Pistacia khinjuk. Journal of Medicinal Plants Research 2011; 5(17):4310-4.
  38. Hassanshahian M, Bayat Z, Saeidi S, Shiri Y. Antimicrobial activity of Trachyspermum ammi essential oil against human bacterial. International Journal of Advanced Biological and Biomedical Research 2014; 2(1):18-24.
  39. Mohsenipour Z, Hassanshahian M. The inhibitory effect of Thymus vulgaris extracts on the planktonic form and biofilm structures of six human pathogenic bacteria. Avicenna J Phytomed 2015; 5(4):309-18.
  40. Bisignano C, Filocamo A, Faulks RM, Mandalari G. In vitro antimicrobial activity of pistachio (Pistacia vera L.) polyphenols. FEMS Microbiol Lett 2013; 341(1):62-7.
  41. Sagdic O, Karahan A, Ozcan M, Ozkan G. Note: effect of some spice extracts on bacterial inhibition. Food Sci Tech Int 2003; 9(5):353-8.
  42. Burt S. Essential oils: their antibacterial properties and potential applications in foods--a review. Int J Food Microbiol 2004; 94(3):223-53.
  43. Allan G, Murghy MJ, Srivastava A, Cowan RA, O'Reilly DJ. Clinical Biochemistry: An Illustrated Colour Text. 2th ed. UK: Churchill Livingstone; 1999.
  44. Hosseini MH, Razavi SH, Mousavi MA. Antimicrobial, physical and mechanical properties of chitosan‐based films incorporated with thyme, clove and cinnamon essential oils. Journal of Food Processing and Preservation 2009; 33(6):727-43.
  45. Knowles JR, Roller S, Murray DB, Naidu AS. Antimicrobial action of carvacrol at different stages of dual-species biofilm development by staphylococcus aureus and salmonella enterica serovar typhimurium. Appl Environ Microbiol 2005; 71(2): 797–803.
  46. Kouidhi B, Al Qurashi YM, Chaieb K. Drug resistance of bacterial dental biofilm and the potential use of natural compounds as alternative for prevention and treatment. Microb Pathog 2015; 80:39-49.
  47. Pratt LA, Kolter R. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol Microbiol 1998; 30(2):285-93.
  48. Mohsenipour Z, M Hassanshahian M. Inhibitory effects of Tamarix hispida extracts on planktonic form and biofilm formation of six pathogenic bacteria, Biol J Microorgan 2015; 4 (13): 25-36.