ANTHELMINTIC ACTIVITY OF BLACK MANGROVE OF AEGICERAS CORNICULATUMHTML Full Text
ANTHELMINTIC ACTIVITY OF BLACK MANGROVE OF AEGICERAS CORNICULATUM
J. Raju* and L. R. Patro
Siddhartha Institute of Pharmacy, Narapally, Hyderabad - 500088, Telangana, India.
ABSTRACT: This study evaluated the scientific basis for the traditional use of Aegiceras corniculatum (Myrsinaceae) stem extract as an anthelmintic agent. The study design were investigation of the traditional anthelmintic medicinal plant Aegiceras corniculatum using in vitro anthelmintic properties of four extracts of the plant were evaluated using earthworms. The earthworm (Pheretima posthuma) resembles both anatomically and physiologically to the intestinal roundworm parasites of human beings. The four crude stem extracts of Aegiceras corniculatum were Petroleum Ether extract (50 &100 mg/ml), Chloroform extract (50 &100 mg/ml), Methanol extract (50 &100 mg/ml) and Aqueous extract (50 &100 mg/ml). The paralysis time of Petroleum Ether extract (96 ± 6.33 &76 ± 2.31), Chloroform extract (116 ± 4.36 & 92 ± 7.50), Ethanol extract (62 ± 3.20 & 34 ± 3.52) and aqueous extract (200 ± 2.22 & 180 ± 3.19) were compared to the standard drug Albendazole suspension 100mg/5ml (5 ± 1.00). The results shown that the four extracts produced dose dependent and significant anthelmintic activities. From these findings the four extracts of Aegiceras corniculatum are rich source of naturally occurring anthelmintic activity. Further work is recommended to evaluate the in vivo anthelmintic activity and toxicity of the extracts.
Aegiceras corniculatum, Earthworms, Anthelmintic activity
INTRODUCTION: Helminth infections resulting to diseases such as ascariasis, hookworm infection and schistosomiasis constitute the bulk of the 13 diseases classified as neglected tropical diseases (NTDs) by the WHO 1. These incapacitating diseases have continued to inflict severe disability and often deaths. It is more pronounced among the impoverished population living in marginalized areas of the world 2. In most developing countries, intestinal helminth infections are a major health concern because factors that pre-dispose humans to these infections abound in these areas.
Children, especially those at a preschool age have been identified as the most vulnerable group with very high rates of infection. Due to the asymptomatic nature of these diseases, the helminths remain undetected and children born in an endemic region may harbour the worms for the most part of their lives3. The manifestation of most parasitic diseases is due to the host responses to the presence of the parasite 4. Also worth considering is the fact that the immune response triggered by helminth infection may drain the body’s ability to fight other diseases, making affected individuals more prone to co-infection 5.
Chemical control of helminthes coupled with improved management has been the important worm control strategy throughout the world. However, development of resistance in helminthes 6 - 7 against conventional anthelmintics is a foremost problem in treatment of helminthes diseases. Henceforth it is important to look for alternative strategies against gastrointestinal nematodes, which have led to the proposal of screening medicinal plants for their anthelmintic activity. Mangrove forests are a special type of vegetation found in the coastal regions of the tropical and subtropical parts of the world. Global area that comprises mangrove forest is about 181000 square km. Mangrove forests are still quite unfamiliar to a vast population due to their limited distribution. However, the people inhabiting areas near mangrove forests heavily depend on these forests to meet their needs including their healthcare. During the early stage of human civilization, mangrove forests drew very little or no attention.
This is to some extent because of the difficulty to access these areas. As the population continued to grow, people had to find new and unexplored sources including mangrove forests 8. Like other terrestrial plants, many mangrove plants have ethnopharmacological relevance and have also been exploited by the local people in the search for remedies for various ailments. However, only a few of the mangrove plants have so far been included in any books listing medicinal plants. This may be due to the difficulty in collecting and identifying these plant species and lack of adequate information available about their uses 9.
The preliminary studies of A. corniculatum bark showed the presence of alkaloids, glycosides, steroids, flavonoids, saponins and tannins 10. The therapeutic applications of the black mangrove like Aegiceras corniculatum (Linn.) (Myrsinaceae) distributed in coastal and estuarine areas of India are well studied. Also, the ethnopharmacological consequence pointed out that the mangrove plants are traditionally used for the treatment of rheumatism, painful arthritis, inflammation, asthma antioxidant, free radical scavenging, anti-inflammatory, antinociceptive, diabetes and hepatoprotective actions 11. However, there are no proper scientific reports available regarding the effects of Aegiceras corniculatum on Diabetes mellitus. Mangroves and associated plants provide a wide domain for therapeutic application in recent years, most yet to be explored. The leaves of A. corniculatum are reported that it have rich in flavonoids with proven anti-inflammatory and antioxidant property 12 - 13. Aegiceras has been used as traditional medicine for years with array of biological activity such as antioxidant, antibacterial, antifungal, antiulcer, anticancer, antiplasmodial and antitumour properties 14 - 15.
MATERIALS AND METHODS:
Plant Material Collection: Aegiceras corniculatum stem were collected from the coringa mangrove forest, Andhra Pradesh and authenticated by the taxonomist, Department of Botany, Andhra University in the month of July 2016 and the specimen voucher (AC01/2016) were preserved in the herbarium.
Plant Extract Preparation: Aegiceras corniculatum stem was dried under shade then made into coarse powder and was weighed for 250 gm and it was soaked in 1litres of Petroleum ether by maceration. The mixture were kept for 3 days in tightly sealed vessels at room temperature and stirred several times daily with a glass rod. This mixture was filtered through Whatman No-1 filter paper. The extract was subjected to Rotavapour evaporation to remove the solvent. The same procedures were used for the chloroform, methanol and aqueous extraction. The extracts obtained were kept in dessicator containing calcium carbonate.
These extracts were stored at - 4 °C until use and dissolved in normal saline on the day of the experiments for the purpose of evaluating pharmacological activity.
Parasites: Adult earthworms (Pheretima posthuma) of 8-10cm in length were used to evaluate in vitro anthelmintic activity 16.
Experimental Treatments: The earthworms were divided into six groups containing six earthworms in each group. Since the earthworm resembles both anatomically and physiologically to the intestinal roundworm parasites of human beings, Adult earthworms (Pheretima posthuma) were used to study anthelmintic activity. All the earthworms were washed in normal saline solution before they were released into 10ml of respective formulation as follows: Vehicle (5% DMF in 10ml normal saline) used as control, Albendazole oral suspension (100 mg/5ml) used as standard. The selected amount of stem extracts of Aegiceras corniculatum were weighed and taken into the test tubes containing 10ml of normal saline. The extracts were Petroleum Ether extract (50 &100 mg/ml), Chloroform extract (50 &100 mg/ml), Methanol extract (50 &100 mg/ml) and Aqueous extract (50 &100 mg/ml). All the extracts and the standard drug solution were freshly prepared before starting the experiments. Diﬀerent extracts and the standard drug solution were poured in diﬀerent petridishes. Observations were made for the time taken to paralyze (Paralysis is when the worm did not revive even in normal saline) and Death (Death is when the worms lost their motility followed with their body colors fading away) the results of each petridish were recorded 17-20.
Statistics: All the results was expressed as mean ± SEM and followed by student ‘t’ test (n = 6)
RESULTS AND DISCUSSION: The crude extracts samples, which were used to evaluate anthelmintic activity, showed variable times at different concentrations and the mean time values were calculated for each parameter. The crude extracts of ethanol showed the significant anthelmintic effect causing death of the worm at all the concentrations but the time of death was different in each case. However, when observed the response of worms in case of paralysis, there was significant variation among the results produced by the different extracts at different concentrations like 25, 50, 100, and 200mg/ml. The ethanol extract showed more significant effect on paralyzing the worms, in terms of paralysis time, at every concentration compared to that of aqueous extracts.
TABLE 1: AN EFFECT OF A. CORNICULATUM PET. ETHER AND CHLOROFORM EXTRACTS ON EARTHWORMS
|Pet. ether extract||Chloroform extract|
|Extracts (mg/ml)||Paralysis time (min)||Death time(min)||Paralysis time(min)||Death time(min)|
|50||96 ± 6.33||125 ± 7.03||116 ± 4.36||138 ± 2.62|
|100||76 ± 2.31||98 ± 1.21||92 ± 7.50||126 ± 7.89|
|Albendazole Suspension100mg/5ml||5 ± 1.00||23 ± 6.34|
Each value represents mean ±SEM (N=6).
TABLE 2: AN EFFECT OF A. CORNICULATUM AQUEOUS AND ETHANOL EXTRACTS ON EARTHWORMS
|Aqueous extract||Ethanol extract|
|Extracts (mg/ml)||Paralysis time (min)||Death time(min)||Paralysis time(min)||Death time(min)|
|50||200 ± 2.22||240 ± 2.12||62 ± 3.20||84 ± 8.29|
|100||180 ± 3.19||210 ± 3.62||34 ± 3.52||56 ± 4.36|
|Albendazole Suspension100mg/5ml||5 ± 1.00||23 ± 6.34|
P<0.001 significantly different compared with reference compound, Albendazole, student’s t-test.
The four crude stem extracts of Aegiceras corniculatum were Petroleum Ether extract (50 & 100 mg/ml), Chloroform extract (50 & 100 mg/ml), Methanol extract (50 & 100 mg/ml) and Aqueous extract (50 & 100 mg/ml). The paralysis time of Petroleum Ether extract (96 ± 6.33 &76 ± 2.31), Chloroform extract (116 ± 4.36 & 92 ± 7.50), Ethanol extract (62 ± 3.20 & 34 ± 3.52) and aqueous extract (200 ± 2.22 & 180 ± 3.19) were compared to the standard drug Albendazole suspension 100mg/5ml (5 ± 1.00). Results were tabulated in Table 1 and 2. Mangroves and associated plants provide a wide domain for therapeutic application in recent years, most yet to be explored. The leaves of A. corniculatum are reportedly rich in flavonoids with proven anti-inflammatory and antioxidant property 12.
CONCLUSION: The results of the present study clearly indicated that the crude methanol extract of A. corniculatum produced significant anthelmintic activity against Indian earthworms Pheretima posthuma. Nevertheless, activities of extracts of the plant investigated on the earthworms were lower than that of the reference compound, Albendazole suspension. Thus this plant having a potential novel and cheap source of potent anthelmintic agent.
To conclude, in the future studies, there is need for thorough phytochemical, clinical and possible studies on molecular mechanism of action. At the same time efforts should be made to standardize the plant extract and formulate the best alternative herbal preparations to replace or complement the synthetic drugs which are in current use.
ACKNOWLEDGEMENT: The authors are thankful to the Department of Botany, Andhra University. Hearty thankful to the management of Siddhartha Institute of Pharmacy, Narapally, Hyderabad, India for providing facilities to carry out the research work.
CONFLICT OF INTEREST: On behalf of all authors, the corresponding author states that there is no conflict of interest.
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How to cite this article:
Raju NJ and Patro LR: Anthelmintic activity of black mangrove of Aegiceras corniculatum. Int J Pharmacognosy 2017; 4(9): 305-08:.doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.4(9).305-08.
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