Effects of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall on caecal amoebiasis in mice

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Abstract

The anti-amoebic effects of crude methanol extracts of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall against Entamoeba histolytica infecting the caecum of mice were studied. Caecal amoebiasis in mice was induced by injection of Entamoeba histolytica trophozoites directly into the caecum. The mice were then treated orally with the extract, a standard drug (metronidazole), or vehicle p.o. for five consecutive days, beginning 24 h after the infection and were examined on the sixth day. At a dose of 1000 mg/kg per day, the extracts of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall had a curative rate of 100, 40 and 26%, respectively. At a concentration of 500 and 250 mg/kg/day, extract from Piper longum fruit was still effective in 93 and 46% of the cases, respectively, while extract from Piper sarmentosum root at a dose of less than 1000 mg/kg per day did not cure any mice from amoebiasis. Extract of Quercus infectoria nut gall at a concentration of 500 and of 250 mg/kg per day cured 26 and 13% of mice, respectively. Metronidazole at a concentration of 125 and of 62.5 mg/kg per day had a curative rate of 100 and 60%, respectively. The severity of caecal wall ulceration was reduced in mice which received the extract and metronidazole as compared to the control animals.

Introduction

Among parasitic infections, amoebiasis ranks third worldwide in lethal infection, after malaria and schistosomiasis (Walsh, 1988, Petri and Mann, 1993). Although it is asymptomatic in 90% of cases, about 50 million people are estimated to suffer from the symptoms of amoebiasis such as haemorrhagic colitis and amoebic liver abscess (Ravdin, 1995). These infections result in 50 000–100 000 deaths annually. In South Africa and India, the disease is rather common (Walsh, 1986).

During 1987–1997, in Thailand, there were more than 50 000 cases of dysentery reported each year and approximately 400 people died. For about 90% of the cases, the cause of dysentery is unknown; however, Entamoeba histolytica was detected in 2–3% of cases (Anon, 1997). Among children under five years who were admitted with acute diarrhea in a hospital, Entamoeba histolytica was confirmed in 7.8% of the cases (Suwatana, 1997). The estimated number of infected cases may be much higher due to the lack of a sensitive and specific diagnostic test (Petri et al., 2000).

The most effective and commonly used drug for treatment of intestinal protozoa infection is metronidazole (Tracy and Webster, 1996). However, this drug has been reported to cause mutagenicity in bacteria (Legator et al., 1975) and is carcinogenic in rodents (Rustia and Shubik, 1972, Shubik, 1972). It has been reported that the human pathogenic bacterium, Helicobacter pylori, becomes resistant to metronidazole in vitro (Zwet et al., 1994). Moreover, it seems to act as an immunosuppressive agent in experimental rats, both in cell-mediated and humoral immune responses (Saxena et al., 1985). These are the main reason why there is a need to develop a safe and effective alternative antiamoebic agent.

For people in developing countries, medicinal plants are popular because their products are safe and widely available at low cost. Some compounds extracted from medicinal plants already play an important role against infectious diseases e.g. quinine from Cinchona sp., and artemisinin from Artemisia annua; both are effective against malaria. In the present study, Piper longum (Linn.) fruit (Plf), Piper sarmentosum (Roxb.) root (Psr) and Quercus infectoria (Oliv.) nut gall (Qin) were selected because these species are routinely used to cure bloody diarrhea in Thai traditional medical practice. It is, therefore, of interest to scientifically evaluate their effect on amoebiasis for potential antiamoebic activity in vivo. The selected plants were extracted with methanol and tested against caecal amoebiasis in mice. The anti-amoebic effect of the extract was compared with the standard drug metronidazole.

Section snippets

Isolation and cultivation of Entamoeba histolytica

The culture of Entamoeba histolytica used in this experiment was isolated from the bloody stool diarrhea of a patient from Maharaj hospital, Nakorn Srithamarat, Thailand. Briefly, the untreated fecal samples were collected after diagnosis as Entamoeba histolytica infection and cultured in Boeck & Drbohlav medium with some modification as described elsewhere (Sawangjaroen et al., 1993). Calf bovine donor serum (10%) was used instead of horse serum. The culture was incubated at 37 °C and Entamoeba

Results and discussion

The effects of extracts from Plf, Psr and Qin against experimental caecal amoebiasis in mice are shown in Table 1. The results from the present study demonstrate that methanol extracts from selected medicinal plants are effective against Entamoeba histolytica in mice as evaluated by the number of mice cured and the reduction of severity of the mice caecal content and caecal wall lesions in comparison to the untreated mice. The anti-amoebic effects of all extracts are clearly dose-dependent.

Acknowledgments

The authors are grateful for financial support by the Faculty of Science, Prince of Songkla University.

References (22)

  • M.S Legator et al.

    Detection of mutagenic activity of metronidazole and niridazole in body fluids of humans and mice

    Science

    (1975)
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