Efficacy of Black Seeds Oil ( Nigella sativa ) against Hymenolepis nana in Infected Mice

Recently, many biological activities (e.g., antioxidant, anti-inflammatory, anticancer, antimicrobial, antifungal and antiparasitic) of Nigella sativa seeds have been reported. We carried out this study to investigate the therapeutic potential of Nigella sativa oil as an alternative and safe treatment against Hymenolepis nana based on an experimental study of white laboratory mice. Twenty-eight Swiss albino mice naturally infected with H. nana were divided into three groups; one group functioned as the control, and the remaining two groups were fed daily doses of black seed oil (2.5 and 5 ml/kg, respectively). We found that the efficacy of the 5 ml/kg Nigella sativa oil dose against H. nana attained 100% 14 days after treatment; the efficacy of the 2.5 ml/kg Nigella sativa oil dose attained an efficacy of 100% 21 days after treatment. Our results indicate that N. sativa oil exhibits significant efficacy against H. nana in infected mice.


INTRODUCTION
Hymenolepis nana, generally known as the dwarf tapeworm is a globally widespread zoonosis disease. Also, it is one of the most common cause of cestode infections, commonly infects rodents as well as human beings [1][2][3]. Mostly, H. nana infection has a cosmopolitan distribution

Short Communication
with the highest prevalence and heaviest parasite burden among children in warm, arid climates with poor sanitation conditions [4,5].
Synthetic anthelmintics are the most effective way of controlling parasitic infections. However, these medicines are costly and sometimes unavailable to smallholder farmers and pastoralists in developing countries. Moreover, there are cases of increased resistance to anthelmintics worldwide in animals [6][7][8].
Medicinal plants could be a provenance of new antiparasitic medicine with high efficacy, low toxicity and lower price [9]. At present, about 80% of the drugs which used around the world produced from natural products or some derivatives inspired by natural precursors [10]. Consequently, plants form a very rich source of bioactive chemical compounds against many diseases [11]. There are many studies on several plants to test their anthelmintic efficacy [12][13][14][15].
Nigella sativa (Family: Ranunculacea), generally recognized as black seed or black cumin or kalonji seed also renowned as habat ul Barakah, is an annual plant growing in Mediterranean countries. It has been traditionally used in Arabian countries, Europe and the Indian subcontinent for dietary and medicinal purposes as a natural remedy for a various of cases and illnesses that include inflammation, diabetes, hypertension, headache, cough, bronchitis, asthma, eczema, influenza, fever and dizziness [16]. Moreover, black seed (N. sativa) was used traditionally for increasing the milk production, as diuretic, appetizing, regulation of menstruation and becoming healthy. In addition, it is added to homemade pastry for decoration and booster of taste [17] also, it could be used as a natural growth promoter [18]. Furthermore, black cumin seeds and its essential oil have been widely used in nutraceuticals, functional foods and pharmaceutical products [19]. Recently, many biological activities of N. sativa seeds have been reported, including: antioxidant, antiinflammatory, anticancer, antimicrobial, antifungal and antiparasitic activities [20][21]. Abdel Daim and Ghazy [22] have revealed the preventive role of N. sativa oil against the toxic effects and it potent antioxidant activities. Also, Hassanien et al. [19] found that N. sativa oil showed stronger antioxidant potential in comparison with synthetic antioxidants. Nigella sativa seeds were reported to contain fixed and volatile oils [23]. Hassanein et al. [24] found that N. sativa seed oil to be rich in oleic and linoleic acids. Al-Naqeep et al. [25] reported N. sativa seeds to contain high amount of oil (30-48%) and the major unsaturated fatty acids were linoleic acid.

Black Seed Oil
Amazing Herbs™ Premium Black Seed Oil (100% Pure Cold-Pressed Black Seed Oil).

Animals
Twenty eight Swiss albino mice of aged 2 months and weight between 25-35 gm each, proven to be naturally infected with Hymenolepis nana (by detection of eggs in fecal samples smeared on microscopic slides), were obtained from the animal facilities of King Saud University, Riyadh, Saudi Arabia. The mice were bred under specified pathogen-free conditions and fed with feed (P 684) of the General Organization for Grain Silos and Flour Mills production in Riyadh, Kingdom of Saudi Arabia. The experiments were approved and followed Saudi Arabian rules for animal protection.

Experimental Design
Animals were divided into three groups, with seven animals in each group.

Control groups
One group functioned as the control group which have animals infected with H. nana untreated.

Experimental group
Two groups functioned as the experimental groups.
Groups 2 and 3 were fed daily by single oral gavage with black seed oil (N. sativa oil) (2.5 and 5 ml/kg) respectively [27], for twenty one days throughout the experimental period [35].

Parasitological Studies
Fresh faecal samples of mice were collected from cages on a day (pre-treatment period) and on day 1, 7, 14, and 21 (post-treatment period).

Statistical Analysis
Results were reported as mean ± SD for each group. Statistical analysis was performed with student's t-test using a Microsoft Excel 2010. All P<0.05 was considered as significant for all statistical analysis in this study.

RESULTS AND DISCUSSION
The present study was constructed to evaluate the effect of N. sativa oil against H. nana in infected mice. The results displayed in Table 1 showing the effects of N. sativa oil against H. nana in infected mice. It is shown the effects of treatment with N. sativa oil on the number of eggs output in fecal pellets of treated infected groups comparing with control group. The results revealed that N. sativa oil has lead to significant decline the mean number of eggs per gram faeces from 583±37.16 to 65±18.7 by 7 th day after treatment with 5 ml/kg with efficacy of 88.85% and reached 100% by 14 th day after treatment. However, treatment with 2.5 ml/kg contribute to decrease of eggs in faeces with efficacy 100% on day 21 after treatment. Moreover, both doses 5 and 2.5 ml/kg lead to disappearance of worms from the intestines of mice by day 14 and 21, respectively. However, that the adverse effects were not noted. Some herbals and plants can be a new source of medicines to treatment parasitic infections with low toxicity and high efficacy [9]. In addition, most drugs that used over the world create from natural materials specially plants due to contain chemical compounds which have bio-activity against several illness [10][11].
In present study N. sativa oil was used as alternative drug to treatment to H. nana in infected mice. The result show that treatment with N. sativa oil lead to highly reduce eggs passed per gram of faeces from one day until 14 day after treatment with efficacy14.24%, 88.85% and 100% respectively, by using does 5ml/kg. However, using dose 2.5ml/kg show efficacy reached to 100% by day 21. Our finding is in agreement with Ayaz et al. [17] who found N. sativa oil has reduced H. nana eggs starting from second day of treatment. Furthermore, the present results were supported by recent studies which reported that N. sativa has a significant efficacy against parasitic worms. Aboul-Ela [26] reported that N. sativa crushed seeds stimulate an oxidative stress against adult worms which was indicated by a reduction in the activities of both antioxidant enzymes and enzymes of glucose metabolism. This perturbation of such enzyme activities in adult worms could render the parasite vulnerable to damage by the host and may play a role in the anti-schistosomal strength of N. sativa .In addition, the extract of N. sativa was reported to work as protective agents against the chromosomal aberrations produce in mouse cells as a result of schistosomiasis. Moreover, Mahmoud et al. [27] found that N. sativa oil has decreased the number of Schistosoma mansoni worms in the liver and reduce the total number of ova stabilized in both the liver and the intestine of infected mice. Furthermore, N. sativa seeds demonstrated an inhibitory effect on egglaying of adult female worms and also exerted active biocidal effects against miracidia, cercariae, and adult worm stages of Schistosoma mansoni [28]. Additionally, N. sativa oil prevented most of the hematological and biochemical changes caused by schistomiasis and significantly improved the antioxidant capacity of Schistosoma mansoniinfected mice [29]. In another study, Kalonji (Black seed) was reported to possess significant efficacy against fascioliasis in buffalos. Kailani et al. [36] found that treatment with kalonji exerted lead to decreasing eggs per gram faeces by 88.2%. In addition, other studies found that N. sativa oil had anthelmintic effect in the rats infected with Trichinella spiralis infection and increased the production of antibodies generated during life cycle of T. spiralis [20]. Moreover, N. sativa oil reduce infection of Aspiculuris tetraptera and its eggs significantly in mice [17].

CONCLUSION
In conclusion, the results of the present study reinforced the effectiveness of N. sativa oil as effective treatment against H. nana through experimental study on the white laboratory mice.

CONSENT
It is not applicable.

ETHICAL APPROVAL
Author hereby declare that "Principles of laboratory animal care" (NIH publication No. 85-23, revised 1985) were followed, as well as specific national laws where applicable. All experiments have been examined and approved by the appropriate ethics committee.