In vitro the Antiprotozoal Activity of Zingibar officinale Alcohol Extract and Metronidazole in Entamoeba gingivalis which Isolated from Patients with Periodontal Disease

Entamoeba gingivalis is parasite found in mouth, causes dental disease. This parasite discovered in dental plaque, the last studies found relation between E. gingivalis and periodontal disease. In present study collected 30 samples collected from patient with age (20-30) years, result show infection with this parasite 46.6%. These samples are cultured in TYSGM-9 media, and then treated with different concentration of alcoholic extract ginger and metronidazole which show inhibited growth of parasite at (25, 50, 75,100 μg/ml. )


Introduction
Periodontitis have different forms of the generality oral disease.About 5% to 20% of people are affected by intense periodontitis.(Burt, 2005) the Parasite has not an intermediate host to infect the host and transmitted through contamination of food and drinks (Idowu et al, 2006).
Study was conducted show that Entamoeba gingivalis was only protozoan found in saliva and plaque.(Lucht et al, 1998) Metronidazole effects on different protozoans like Entamoeba histolytica and most Gram-negative and Gram-positive anaerobic bacteria.(Collin et al, 1997) E. gingivalis is oversensitive to Metronidazole.(Azzouni et al, 1994) Ginger is the rhizomes of the Zingiber officinale Roscoe; it's used as spice and medicine for some cases like diarrhea, headaches, cold, stomach disorder, nausea and others cases.(Baliga et al, 2011;Baliga et al, 2012;Haniadka et al, 2013) it also has strong anxiolytic and activities against emetic.(Vishwakarma et al. 2002).this herbal is safety for many people especially pregnant women.Clinical tests prove ginger have effect against chemotherapy like vomiting and nausea.(Westfall et al, 2004)

Material And Method
A/ sample collection: 30 samples collected from Private clinic by using sterile cotton swap from patients with periodontitis to detect Entamoeba gingivalis.B/ Isolation procedures a / examine under microscope: Used wet mount and normal saline to detect parasite and examined directly under microscope.(Onyido et al, 2011).

C/ Prepared extract of ginger and metronidazole:
Ginger powder gets from market (50gm).Mix powder with 100ml ethanol (99.9%) in an electric blender for 30 min.This suspension was filtered.Then methanol was removed in a rotary evaporator to produce a dry powder.To obtain concentrations 12.5μg/ml, 25μg/ml, 50μg/ml and 100 μg/ml, powder was dissolved in ethanol (Naz et. al., 2007).

Result
30 samples were collected from patients aged 20 to 30 years, found 46.6% was positive with Entamoeba gingivalis.All samples were positive when cultured in TYSGM-9.
In present study was investigated in vitro effect of Ethanol Extract of ginger (Zingiber officinale) on the growth and motility of Entamoeba gingivalis compared to the standard drug metronidazole with significant at the 0.05 level Growth of parasite show in figures 1 and 2. Take drop of broth and calculate dead E. gingivalis / 100, which show 53% of E. gingivalis was killed with ethanol extract of ginger and 89% of this parasite killed with metronidazole.

Discussion
The trophozoite of Entamoeba gingvalis ranges in size from 8 to 20 μm and morphologically resembles that of E. histolytica found in mouth .In present study samples take from people whom have from continue inflammation in periodontal are more infection with parasite especially Entamoeba gingivalis.In this study show rate of infection with parasite among people with age 20-30years, which was 46.6%, this result was closely to another study in Babylon city rate of infection with E. gingivalis was 42.9 % ( Al-hamiary et al., 2011).while31.67%rate of infection with parasite in another study (Onyido et. al., 2011) in culture noted all of them are grow in 37C˚ for 48-72 hours (Gannon JT and Linke H A, 1990).Cause of infection with E. gingivalis possibly to no oral hygiene (Onyido et al., 2011) or this parasite is opportunists (Talaro and Talaro, 2002).After cultured parasite and deal with extract of ginger, noted growth of parasite was inhibition in 100 μg/ml (0.045) in compare with metronidazole 100 μg/ml (0.087) (figure1and2) with significant difference.This result show the effect of ginger on growth of parasite like another research which proves the effect of ginger as inhibited for parasite and microorganisms (Aly and Mantawy , 2013).In other study, used ginger against Haemonchus contortus which killed all of these worms (Qbal et al., 2001).We don't found any research conformity with this study.
Metronidazole was killed E. gingivalis and this result was similar to another result in other research which insures Metronidazole killed the parasite (Eloufir et. al., 2014) while other study prove metronidazole treat amoebiasis (Klasco ,2006).