In Vitro Antileishmanial Activity of Achillea santolina Essential Oil against Leishmania infantum Promastigote by Methylthiazole Tetrazolium (MTT) and Trypan Blue Colorimetric Methods

Document Type : Original Articles


1 Department of Pathobiology, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Parasitology, Faculty of Veterinary, Urmia Branch, Islamic Azad University, Urmia, Iran


Leishmaniasis causes parasitic infections, especially in developing countries. The disease has not yet been controlled because of the absence of an effective vaccine and low-cost treatment. Achillea santolina essential oil (ASEO) might control the disease as it has antimicrobial properties. This study investigated the in vitro antileishmanial activity of ASEO against Leishmania infantum promastigote using the methylthiazole tetrazolium (MTT) and trypan blue colorimetric methods. The standard strain of L. infantum (MCAN/IR/96/LON49) promastigotes was prepared and cultured in a 96-well Novy-MacNeal-Nicolle (NNN) medium. The effects of different concentrations of saline, ASEO, and glucantime (10, 50, 100, 200, 500, and 1000 mg/mL) were examined in 24-, 48-, and 72-hour intervals using the MTT and trypan blue test methods.The use of ASEO reduced viability in all concentrations compared to the control group in times of 48 (p<0.05) and 72 h (p<0.05). Treatment with glucantime and ASEO had similar efficiency with the concentration of 1000 mL/mg in both methods after 72 h. The results showed that viability was significantly lower in the ASEO group with increases in time using both methods (p<0.05). Cohen’s Kappa coefficient showed a significant agreement between the obtained results for the two methods (Kappa=0.856; p<0.001).In sum, the results showed in vitro antileishmanial activity of ASEO, but more clinical studies are needed to confirm the efficiency. ASEO can be used as an agent and/or in combination with synthetic agents for the treatment of leishmaniasis disease.


Main Subjects

Article Title [French]

Activité Antileishmanienne In vitro de l'huile Essentielle d'Achillea santolina contre Leishmania infantum Promastigote par les Méthodes Colorimétriques au Méthylthiazole Tétrazolium (MTT) et au Bleu Trypan

Abstract [French]

La leishmaniose provoque des infections parasitaires, en particulier dans les pays en développement. La maladie n'a pas encore été maîtrisée en raison de l'absence d'un vaccin efficace et d'un traitement peu coûteux. L'huile essentielle d'Achillea santolina (ASEO) pourrait contrôler la maladie car elle possède des propriétés antimicrobiennes. Cette étude a examiné l'activité antileishmanienne in vitro de l'ASEO contre le Promastigote de Leishmania infantum en utilisant les méthodes colorimétriques au méthylthiazole tétrazolium (MTT) et au bleu trypan. La souche standard de promastigotes de L. infantum (MCAN/IR/96/LON49) a été préparée et cultivée dans un milieu 96 puits Novy-MacNeal-Nicolle (NNN). Les effets de différentes concentrations de solution saline, d'ASEO et de glucantime (10, 50, 100, 200, 500 et1000 mg/mL) ont été examinés à des intervalles de 24, 48 et 72 heures à l'aide des méthodes de test MTT et bleu trypan. L'utilisation d'ASEO a réduit la viabilité à toutes les concentrations par rapport au groupe témoin dans des délais de 48 (p<0.05) et 72 h (p<0.05). Le traitement au glucantime et à l'ASEO a eu une efficacité similaire avec la concentration de 1000 mL/mg dans les deux méthodes après 72 h. Les résultats ont montré que la viabilité était significativement plus faible dans le groupe ASEO avec des augmentations de temps en utilisant les deux méthodes (p<0.05). Le coefficient Kappa de Cohen a montré une concordance significative entre les résultats obtenus pour les deux méthodes (Kappa=0.856; p<0.001). En somme, les résultats ont montré une activité antileishmanienne in vitro de l'ASEO, mais d'autres études cliniques sont nécessaires pour confirmer l'efficacité. ASEO peut être utilisé comme agent et/ou en combinaison avec des agents synthétiques pour le traitement de la leishmaniose.

Keywords [French]

  • activité antileishmanienne
  • Leishmania Infantum
  • Achillea santolina
  • méthylthiazole tétrazolium
  • méthode au bleu trypan
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