Antischistosomal activity of ginger (Zingiber officinale) - Springer Link

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Feb 16, 2011 - Antischistosomal activity of ginger (Zingiber officinale) against Schistosoma mansoni harbored in C57 mice. Osama M. S. Mostafa & Refaat A.
Parasitol Res (2011) 109:395–403 DOI 10.1007/s00436-011-2267-x

ORIGINAL PAPER

Antischistosomal activity of ginger (Zingiber officinale) against Schistosoma mansoni harbored in C57 mice Osama M. S. Mostafa & Refaat A. Eid & Mohamed A. Adly

Received: 11 January 2011 / Accepted: 18 January 2011 / Published online: 16 February 2011 # Springer-Verlag 2011

Abstract The repeated chemotherapy of schistosomiasis has resulted in the emergence of drug-resistant schistosome strains. The development of such resistance has drawn the attention of many authors to alternative drugs. Many medicinal plants were studied to investigate their antischistosomal potency. The present work aimed to evaluate antischistosomal activity of crude aqueous extract of ginger against Schistosoma mansoni. Sixteen mice of C57 strain were exposed to 100±10 cercariae per mouse by the tail immersion method; the mice were divided into two groups: untreated group and ginger-treated one. All mice were sacrificed at the end of 10th week post-infection. Worm recovery and egg counting in the hepatic tissues and faeces were determined. Surface topography of the recovered worms was studied by scanning electron microscopy. Histopathological examination of liver and intestine was

done using routine histological procedures. The worm burden and the egg density in liver and faeces of mice treated with ginger were fewer than in non-treated ones. Scanning electron microscopical examination revealed that male worms recovered from mice treated with ginger lost their normal surface architecture, since its surface showed partial loss of tubercles’ spines, extensive erosion in intertubercle tegumental regions and numerous small blebs around tubercles. Histopathological data indicated a reduction in the number and size of granulomatous inflammatory infiltrations in the liver and intestine of treated mice compared to non-treated mice. The results of the present work suggested that ginger has antischistosomal activities and provided a basis for subsequent experimental and clinical trials.

O. M. S. Mostafa : M. A. Adly Biology Department, Faculty of Science, King Khaled University, Abha P.O. Box 9004, Saudi Arabia

Introduction

M. A. Adly e-mail: [email protected] O. M. S. Mostafa (*) Zoology Department, Faculty of Science, Ain Shams University, Abbassia, 11566, Cairo, Egypt e-mail: [email protected] R. A. Eid Faculty of Medicine, King Khaled University, Abha P.O. Box 9004, Saudi Arabia e-mail: [email protected] M. A. Adly Zoology Department, Faculty of Science, Sohag University, 82524, Sohag, Egypt

Ginger (Zingiber officinale) is a perennial plant with narrow, bright green, grass-like leaves and yellowish green flowers with purple markings. Ginger is cultivated in the tropics for its edible rhizome which used for a variety of purposes, including culinary and medicinal (Grant and Lutz 2000). The efficacy of ginger is purported to be a result of its aromatic, carminative and absorbent properties (Govindarajan 1982). The medicinal properties attributed to ginger include anti-arthritic (Srivastava and Mustafa 1989; Bliddal et al. 2000), anti-migraine (Mustafa and Srivastava 1990; Cady et al. 2005), anti-thrombotic (Bordia et al. 1997; Thomson et al. 2002), anti-inflammatory (Thomson et al. 2002; Penna et al. 2003), hypolipidaemic (Bordia et al. 1997; Thomson et al. 2002; Bhandari et al. 2005; Al-Amin et al. 2006),

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Parasitol Res (2011) 109:395–403

Table 1 Worm recovery and egg density in the hepatic tissues of C57 mice infected with S. mansoni, treated or nontreated with ginger

Worm recovery (mean±SE)

Non-treated Treated

Eggs/gm of liver (mean±SE)

Male

Female

Total

14.3±1.2 7.6±0.9

8.7±1.7 3.4±0.66

23±1.3 11±0.7

hypocholesterolaemic (Bhandari et al. 2005; Fuhrman et al. 2000), anti-nausea properties (Ernst and Pittler 2000; Portnoi et al. 2003) and anti-diabetic (Al-Amin et al. 2006). Few investigations were done upon the anthelmintic activity of ginger and its constituents. Iqbal et al. (2006) showed that crude powder and crude aqueous extract of dried ginger possessed anthelmintic activity in sheep. Lin et al. (2010a) suggested that the 6-gingerol, 10-shogaol, 10gingerol, 6-shogaol and hexahydrocurcumin, a constituent isolated from the ginger might be used as larvicidal agents against Angiostrongylus cantonensis. Lin et al. (2010b) investigated the anthelmintic activity of previous compounds against the nematode worm Anisakis simplex; they reported that these compounds kill or reduce spontaneous movement in A. simplex larvae. Concerned with schistosomiasis, the bioactivity of an ethyl acetate extract of ginger (Z. officinale) towards Schistosoma mansoni adult pairs, both cultured in vitro and in vivo in laboratory mice, was investigated. In vitro, the results are promising but in vivo no significant difference between treated and non-treated mice was observed (Sanderson et al. 2002). This work aimed to evaluate antischistosomal activity of crude aqueous extract of ginger against S. mansoni in C57 mice by assessment of five parameters, worm recovery, egg density in the liver of infected mice, eggs passed out with faeces, surface topography of the recovered worms, and finally histopathological examination of liver and intestine.

Material and methods Source of parasite and host Biomphalaria alexandrina snails shedding S. mansoni cercariae were supplied by the Schistosome Biological Supply Program at Theodor Bilharz Research Institute, Imbaba, Giza, Egypt. Sixteen adult male mice of C57 strain, weighing 18–22 g each, were obtained from the animal house of Biology Department, Faculty of Science, King Khalid University, Abha, Saudi Arabia. The

5148.3±211.6 4069±98.4

animals were given access to water and standard diet and were monitored daily for health status. Experimental design The mice were divided into two groups: group 1 consisted of eight mice infected and untreated, group 2 consisted of eight mice infected and treated with ginger extract. All mice were sacrificed at the end of 10th week post-infection. Infection of mice The mice were exposed to 100±10 S. mansoni cercariae per mouse by the tail immersion method, modified by Oliver and Stirewalt (1952). Treatment of mice Ginger rhizomes were purchased from local suppliers in Abha City, Saudi Arabia. Aqueous extract was prepared by homogenizing 30 g of fresh ginger rhizomes in 60 ml of distilled water then the ginger paste was squeezed out through a piece of cloth to obtain the extract. The filtrate was stored at −20°C and the extract freshly prepared every 3 days. The dose 500 mg/kg body weight was selected for this work. Ginger extract was given orally with an oesophageal tube every day for 5 weeks from the fifth week post-infection. Worm recovery The recovery of S. mansoni worms from the hepatic portal system and mesenteric veins of sacrificed mice was done by the perfusion technique described by Smithers and Terry (1965). Egg counting in hepatic tissues After scarification of mice, a piece of the liver tissues was stored at −20°C for counting of deposited eggs. The eggs were counted according to the method of Cheever (1968). Egg counting in faeces Faeces was collected from infected mice at the end of fifth, sixth, seventh, eighth and ninth weeks post-infection. The eggs were counted according to the method of Doenhoff et al. (1978).

Table 2 Eggs density (mean±SE) per gram of faeces of C57 mice infected with S. mansoni, treated or non-treated with ginger

Non-treated Treated

Fifth week post-infection

Sixth week post-infection

Seventh week post-infection

Eighth week post-infection

Ninth week post-infection

55±23.6 0

266.6±56.9 0

433.3±67.1 16.6±1.5

513.2±59.4 70.5±17.8

611.4±78.6 130.8±20.9

Parasitol Res (2011) 109:395–403

Fig. 1 Scanning electron micrograph of dorso-ventral surface of male worm recovered from non-treated mice showing numerous large tubercles (black arrow) bearing spines (S) on the dorsal part and numerous rows of minute spines (mS) on the ventral side

Scanning electron microscopical techniques Worms were fixed in 4% glutraldehyde in sodium cacodylate buffer for 2 h, washed in the same buffer at a pH of 7.4, dehydrated with ethanol and critical point dried. Specimens were mounted on stubs, coated with carbon and gold and examined with JOEL-1200EX2 electron microscope at King Khaled University, Abha, Saudi Arabia. Histological methods Tissue samples of the liver and intestine of all groups were immediately fixed after animal dissection in two fixatives; Bouin’s and Carnoy’s fixatives, dehydrated and processed for paraffin sectioning. Sections were then deparaffinized, stained with hematoxylin and eosin double stain and examined under Olympus light microscope. To assess the size of tissue granuloma, the mean diameter (μm) was measured. For each group, 30 granulomas were chosen from different sections and different mice.

Fig. 2 Scanning electron micrograph of dorsal surface of male worm recovered from treated mice showing abnormal surface architecture, erosion in the inter-tubercle area (black arrow), loss of spine and numerous small blebs around tubercles (sB)

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Fig. 3 Scanning electron micrograph of anterior end of male worm recovered from treated mice showing abnormal oral sucker (black arrow)

Statistical analysis Results were subjected to Student’s t test using SPSS program version 8 to determine the significant of data.

Results The mean number of S. mansoni worms recovered from treated group (11±0.73) was lower than that in the infected and untreated one (23±1.38), and the differences between them were significant (P