In vivo evaluation of anti-parasitic effects of Artemisia absinthium extracts on Syphacia parasite
B Esfandiari, M Youssefi, M Keighobadi, H Nahrevanian
Keywords
artemisia, extract, herbs, iran, syphacia
Citation
B Esfandiari, M Youssefi, M Keighobadi, H Nahrevanian. In vivo evaluation of anti-parasitic effects of Artemisia absinthium extracts on Syphacia parasite. The Internet Journal of Parasitic Diseases. 2006 Volume 2 Number 2.
Abstract
Herbs are the common resources for drugs in the both forms of traditional or pure products. WHO prepared a comprehensive list of herbs as well as their extracts are currently using for experimental trials against a variety of diseases. Such herbal products could be used for providing novel effective drug targets. This study has been carried out in order to evaluate the effects of various native herbs are currently used for anti parasitic purposes in Iran as well as their application in traditional medicines. The authors examined direct effects of
Introduction
Using herbs for treatment of diseases returns to thousands years ago; and nowadays, one third of medicines has been estimated to contain natural herbs or their extracts (1). Herbs always account for one of the general medicine drugs because such products could be used for producing very important and effective medicines (2, 3).
Artemisia is a stable and very aromatic plant with 75 cm in length. Its stems covered by a soft fluff coloring grayish having needle leaves with coarse edges. This is the native of Mediterranean area and central Europe (4). In tropical regions, Artemisia grows in margins of roads and overt regions self-growingly with small, globular yellow- greenish flowers which appear in the tops of thin stems during middle summer to middle of the fall.
Syphacia parasite is a nematode worm including two genus Syphacia obvolata and Syphacia murris, living mostly in cecum and colon of mice. This parasite is called mouse pinworm and its size ranging 1.1 to 1.5 mm with direct transmission. There have been reported only three Syphacia infections in Iranian children (5). In regard to the investigations of various native herbs efficacy used for their anti parasitic purposes in Iran as well as using these herbs in traditional medicine; this study was designed to examine direct effects of Artemisia extract in removing Syphacia infection in mouse.
Materials and Methods
Plants and extract preparation
The authors examined direct effects of Artemisia absinthium extract in removing Syphacia parasite in the experimental mouse. Following preparation of Artemisia from Agricultural Faculty of Shiraz University located in 12 km of Shiraz - Isfahan Road, in the Bajgah region, it transferred to the Medical Faculty of Babol. They were then air-dried in the sun shade, mixed using Blender device, drilled and powdered. Herbal extract was prepared using Reflux Condenser by adding 40 g of dried plant weight including leaves and floral top branch trimming of plant to 150 ml absolute ethanol (Merck), mixed well, incubated for 24 h at room temperature (22-25 C) and finally extracted by evaporator rotation device during 5 h.
Parasite
To prepare Syphacia nematode worm, young mice were trapped and spinal cord cut by vivisection from cecum and colon. After washing in normal sera, adult Syphacia was specified; female worms containing ova were identified and isolated from males. Mature female worms were selected and their ova were collected by removing uterus.
Animals and groups
Male Balb/c mice weighing 20-25 g and 4-6 weeks old were purchased from Karaj Laboratory Animal Section of Pasture Institute of Iran. All mice primarily were examined for Syphacia infection by fecal examination of floated samples; all infected mice were exempted form study. Twenty five animals were divided into five groups (n= 5 mice) and were kept at starvation for 12 h before study. Entire groups were fed independently and orally by 200 µL of suspension containing Syphacia ova at concentration of 10 ova in 20 µL of solution.
Herbal treatment
Experimental mice were treated with A. absinthium extract 10 days after infection by orally inoculated with
Results
Microscopical examination of feces indicated no Syphacia ova in all experimental groups, except control mice, which was verified later by histopathological study of target organs at the terminal stage of experiment. Syphacia ova were only observed in feces of control group starting 25 days after infection. Different concentrations of 2.5%, 5% and 10% of Artemisia extract represented high anti-parasitic effects on Syphacia parasite. The efficacy of extract was similar to choice treatment of nematode with Pyrantel-Pamoate. In extract-treated groups, some clinical symptoms were observed at third day of treatment, including confusion, imbalance and ventral fast respiration, which may be side effects of ethanol consumption.
In comparison of number of dead worms among control and test groups, a Statistical Mann-Whithney test was applied. It was indicated a significant difference in control mice when compared with test groups (P<0.05). In fourth group with the highest doses of extract treatment; two mice died at the middle stage of study. This may be a consequence of remaining toxic Comarrin substance (Thujune) in the extract.
Discussion
This study demonstrated that crude extracts of A. absinthium are effective at inhibiting the in vivo growth and development of the Syphacia parasite. Microscopic examination of the fecal samples showed a significant decline of parasite survival. There are some reports are supporting this study. Arab (2006), introduced Artemisia sieberi as one of the most effective candidates against Coccidiosis in the chickens infected with Eimeria tenella and Eimeria acervolina, but not on Eimeria maxima (6). In Dominican region located in the west of India, some worms became resistant against common medicines called Worm Bag; then various herbs or a combination of them used for curing infected people. Artemisia is one of the most common plants used for removing intestinal helminthiasis (7). The natural and synthetic Artemisia was able to act against malaria parasite, especially its drug resistant type of Plasmodium falciparum (8). Moreover, Artemisia can be used against micro organisms including Helicobacter pylori (9).
Therapies were well tolerated however; nausea, vomiting, dizziness, sleep disorders, and other neurological side effects were also reprted (10). The clinical use and the toxicity and teratogenicity of Artemisia and its derivatives, however, raise some queries and require further studies. Nevertheless, the new compounds are of considerable promise and further intense collaboration has now been planned between the WHO and research laboratories (11). Although there have been considerable scientific advances over the past hundred years, the overall burden of plant chemistry is currently increasing. The WHO is recommending new therapies, based on the use of Artemisin derivatives or combination therapy (12). In addition to some of therapeutic effects of Artemisia, modern scientific studies are required to clarify the effective fraction sites of this herb (13).
Correspondence to
Dr. H. Nahrevanian, Department of Parasitology, Pasteur Institute of Iran, Pasteur Avenue, Tehran 13164, Iran Tel / Fax: +98-21-66968855, E-mail: mobcghn@yahoo.co.uk