Antiparasitic Efficacy of Artemisia Ludoviciana Nutt. (Asteraceae) Essential Oil for Acanthamoeba Castellanii, Leishmania Infantum and Trichomonas Vaginalis
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Original Article Antiparasitic Efficacy ofArtemisia ludoviciana Nutt. (Asteraceae) Essential Oil for Acanthamoeba castellanii, Leishmania infantum and Trichomonas vaginalis Ayşe Baldemir1*, Ülkü Karaman2, Selen İlgün1, Gamze Kaçmaz2, Betül Demirci3 1Erciyes University, Faculty of Pharmacy, Department of Pharmaceutical Botany, Kayseri, TURKEY. 2Ordu University, Faculty of Medicine, Department of Medical Parasitology, Ordu, TURKEY. 3Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy, Eskişehir, TURKEY. ABSTRACT Background: Artemisia ludoviciana Nutt. (Asteraceae) is an aromatic, herbaceous, perennial plant and known commonly name as “White Sage”, “Black Sage”, “Prairie Sage” or “Cudweed Sagewort”. It is traditionally used as an antispasmodic, anthelminthic, antidiarrhoeal, stomachic, hepatic colic, appetizer, and regulator of menstruation, antimalaric and antiparasitic efficiancy. Objective: The essential oil composition of the flowering herb of A. ludoviciana (AL) was investigated and for the first time, the oil was screened for antiparasitic activity. Methods: The chemical composition of the hydrodistilled essential oil (EO) obtained from the herb with flowered of A. ludoviciana was analyzed by Gas Chromatography–Flame Ionization Detector (GC-FID) and Gas Chromatography–Mass Spectrometry (GC-MS). Results and Discussion: The outcomes showed that the major components of oil were camphor (40.6%), 1,8-cineole (25.5%) and camphene (4.7%), among 45 identified compounds, comprising 98.5 of the total oil. In addition, the oil was tested against Acanthamoeba castellanii, Leishmania infantum and Trichomonas vaginalis. Conclusion: In this study, it is first demonstrated that A. ludoviciana essential oil (AL-EO) is effective against three important parasites. Key words: Artemisia ludoviciana, Camphor, 1,8-Cineole, Camphene, Antiparasitic effect, GC-FID, GC/MS. Submission Date: 18-08-2017; INTRODUCTION Revision Date: 12-10-2017; Accepted Date: 29-10-2017 Plants/plant extracts and their active com- The genus Artemisia L. comprises important DOI: 10.5530/ijper.52.3.48 ponents have been used for many centuries Correspondence: medicinal plants, which have gained phyto- Ayşe Baldemir, as treatments for diseases from headaches chemical attention due to their biological Assistant Prof., Department to parasite infections. In the last 20-30 years, and chemical diversity, and essential oil of Pharmaceutical Botany, Faculty of Pharmacy, Erciyes researchers have seriously focused on deter- production. Artemisia species, widespread University, 38039 Kayseri, mining whether plant-derived traditional throughout the world, are frequently used TURKEY. Phone: +90-352-2076666 remedies are effective and what their mode for the treatment of diseases such as E-mail: aysebaldemir@gmail. of action is. Several studies proving the malaria, hepatitis, cancer, inflammation and com effects of plants on parasite infections have infections by fungi, bacteria and viruses.2 been undertaken using aqueous or alcoholic The genus Artemisia includes 23 perennial extracts and essential oils (EOs).1 The EOs aromatic herbs and shrubs that grow wild generally have a broad spectrum of bioac- in Turkey.3 In the literature, there are only tivity, owing to the presence of several active a few papers dealing with the essential oil ingredients or secondary metabolites, which composition and properties of Artemisia work through various modes of action.2 ludoviciana Nutt. from among these www.ijper.org 416 Indian Journal of Pharmaceutical Education and Research | Vol 52 | Issue 3 | Jul-Sep, 2018 Baldemir, et al.: Antiparasitic Activity of Artemisia ludoviciana Nutt. Essential Oil species. Previous studies reported that main components aminacrine, sodium edetate and docusate sodium for of leaf oils of A. ludoviciana (AL) grown in USA 2 weeks duration administered by the vaginal route and Mexico analyzed by gas chromatography–mass twice per week.21,22 spectrometry (GC-MS) were alpha-pinene, camphene, Acanthamoeba species may proliferate easily in natural 1,8-cineole, camphor, borneol, nonanal, linalool, carva- environments, infected organs, and on xenic and axenic crol and p-alpha-dimethylbenzyl alcohol.4,5 The dried cultures. Acanthamoeba have been identified in natural whole plant, a leaf infusion and essential oil (EO) of environments like water, thermal water, sea water, soil and A. ludoviciana Nutt. have traditionally been used due air. Additionally they have been reported to be isolated to antispasmodic, anthelminthic, antidiarrhoeal, stomachic, from human-made environments such as drinking hepatic colic, appetizer, regulator of menstruation, anti- water, bottled spring water, distilled water in laboratories, 4,6,7 malarial, and antiparasitic activity. Hexane, acetone, chlorinated swimming pools and contact lens storage methanol and aqueous extracts of A. ludoviciana cases.23-26 were found to be active in vitro against the parasitic 6 Granulamatous amoebic encephalitis (GAE) cases are protozoa Entamoeba histolytica and Giardia lamblia. commonly detected to involve Acanthamoeba culbertsoni, In Africa, Asia, Europe and South and North America, A. castellanii and A. rhysoides species. Acanthamoeba kera- 8 nearly 350 million people are at risk of leishmaniasis. titis is a parasitosis caused by a variety of Acanthamoeba Between 2000 and 2014 in Turkey a total of 413 cases species. Preparatory factors are trauma, contact lens were reported of visceral leishmaniasis (VL) infection use, and contact of infected water with the cornea. It is due to the vector Leishmania infantum. There were observed in healthy individuals and causes severe ocu- 29,845 cases of cutaneous leishmaniasis (CL) due to lar pain, burning, vision disruption and stromal infiltra- the vector Leishmania tropica from 2000-2014.9 Treat- tion with ring shape. Over time vision is impaired and ment of the parasite uses antileishmanial medications the eye may even be lost. In Acanthamoeba keratitis cases, like sodium stibogluconate, miltefosine, parmomycin, most commonly A. castellanii and A. polyphaga are iden- amfotericin B and pentamidine.10 However, the effects tified.27,28 For treatment a variety of medications have of these medications are limited and there are serious been tried. Among effective agents, propamidine isethi- side effects including nephrotoxic, hepatotoxic and onate, ketoconazole, miconazole and itraconazole may teratogenic effects.11,12 Globally and in Turkey, it has been reported that resistance has developed to the primary be listed. Surgical cleaning of the lesion and additionally oral and local administration of miconazole have been medication for treatment of CL and VL of five valu- 29 able antimony species, sodium stibogluconate (Pento- emphasized as effective. stam®) and megluminantiamoniate (Glucantime®).12-16 In this study, EO of the A. ludoviciana flowering herb Due to the lack of an effective prophylactic against the was analyzed with GC and GC-MS and antiparasitic disease, the toxic effects of currently-used medications efficacy of the oil against Acanthamoeba castellanii, Leish- and the increasing resistance to these medications, the mania infantum and Trichomonas vaginalis was examined for necessity for discovery and development of new thera- the first time. peutic agents has been reported.11,17,18 Trichomoniosis is a common infection everywhere in MATERIALS AND METHODS the world and the infection rates are reported to show great variations from country to country and society to Plant material society. Researchers have stated that the different results The flowering herb of A. ludoviciana was obtained from obtained by different people in different regions may be Zeytinburnu Medicinal Plant Garden, Istanbul, Turkey due to factors such as the use of different techniques for in 2016. diagnosis, and deficient and mistaken assessments.19,20 Trichomonas may be treated with metronidazole, Isolation of the essential oil tinidazole, nimorazole, secnidazole and ornidazole. The plant materials were air dried at room temperature It is recommended to administer paired treatment at and were subjected to hydrodistillation for 3 h using the same time. Metranidazole may be administered to a Clevenger-type apparatus to extract essential oils. resistant cases 4-6 weeks after first treatment. Patients The oils were dried over anhydrous sodium sulfate to developing side effects linked to metranidazole may remove moisture and stored at +4°C until analyzed and be given agents like polyoxethylene nonylphenol, tested further. Indian Journal of Pharmaceutical Education and Research | Vol 52 | Issue 3 | Jul-Sep, 2018 417 Baldemir, et al.: Antiparasitic Activity of Artemisia ludoviciana Nutt. Essential Oil Analysis of essential oil Preparation of Medium GC-MS analysis Agar of 1.5 g was heated and dissolved in 100 mL Page The GC-MS analysis was carried out with an Agilent solution, autoclaved for 15 min at 121°C for 15 min and 5975 GC-MSD system. An Innowax FSC column distributed to petri dishes. The cultures were stored at (60 m x 0.25 mm, 0.25 mm film thickness) was used 4°C until use. with helium as carrier gas (0.8 ml/min). The GC Culture oven temperature was kept at 60°C for 10 min and programmed to 220 °C at a rate of 4°C/min, and kept The prepared media were watered with 0.5 mL Page constant at 220 °C for 10 min and then programmed to solution and 24-h E.coli strains were spread on the agar. 240 °C at a rate of 1°C/min. Split ratio was adjusted to Samples taken from A. castalleni strains