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IOSR Journal of Pharmacy
Vol. 2, Issue 3, May-June, 2012, pp.395-397



                 Comparative anthelmintic activity of Aegle marmelos
                               Linn leaves and pulp
                  Kishan Singh*, Krishn Kumar Agrawal, Jeetendra Kumar Gupta
                       Institute of Pharmaceutical Research, GLA University Mathura, (U.P.)-281406


ABSTRACT
        In the present work, experiments were conducted to evaluate the possible anthelmintic effects of different extracts of
Aegle marmelos leaves and pulp on Indian earthworm (Pheretima posthuma) at 20mg/ml concentration. Results were
expressed in terms of time for paralysis and time for death of worms. Piperazine citrate, pyrantal pamoate and albendazole
were used as a standards and DMSO in distill water as a control group. The result revealed that leaves extracts of Aegle
marmelos has better activity that pulp extracts but not better than the standards.

Keywords: Anthelmintic, Albendazole, Aegle marmelos, pyrantel pamoate

INTRODUCTION
          Helminthes parasite infections are global problems with serious social and economic repercussions in the Third
World countries. Parasitic helminths affect animals and man and animals than any single group of parasites, causing
considerable hardship and stunted growth. Most diseases caused by helminthes are of a chronic, debilitating nature; they
probably cause more morbidity and greater economic and social deprivation among humans The diseases affect the health
status of a large fraction of the human population as well as animals.1 Herbal drugs have been in use since ancient times for
the treatment of parasitic disease in human and could be of value in preventing the development of resistance.2, 3 A tree has
unarmed branches and glabrous pedicles in pairs. The fruits are used as laxative and refrigerant. Prunes are often added to
cathartic decoctions, improving their flavor and promoting their effect. They are also given in combination with other drugs
in leucorrhoea and irregular menstruation. Literature survey revealed that plant extract has yet not been screened for
anthelmintic activity. Therefore, the objective of this work was to assess the anthelmintic activity of Aegle marmelos leaves.
Material and Methods
Drugs and chemicals: Piperazine citrate, albendazole and pyrantal pamoate were used as standard drugs.
Chemicals: Petroleum ether (60-80), Ethanol, Di methyl sulfoxide (DMSO) and sodium chloride were used in various part of
experiment.
Collection of plant material: Leaves of Aegle marmelos were collected from local area of Mathura and authenticated by
Birbal Sahani Institute of Palaeobotony, Lucknow, Uttar Pradesh. (Voucher specimen no.13371/2011). Collected leaves were
washed and dried at room temperature. After complete drying, leaves were powdered and passed through a sieve and stored
in air tight container.
Preparation of extracts: The Aegle marmelos were collected from Mathura (Uttar Pradesh) and dried in shade and coarsely
powdered. It was than passed through the sieve no. 20. A weighted quantity (460g) of the powder drug was extracted with
petroleum ether (60-800C) using soxhlet extractor. Defatted drug was subjected to ethanolic extraction and extract was dried
by distilling off the solvent and then dried in desiccator. The marc collected after ethanolic extraction was subjected to
aqueous extraction by maceration process for seven days consecutively and then extract was dried by evaporating the water
and stored for further activity.
Experimental worms: The earthworms of 3-5 cm in length and 0.1-0.2 cm in width were used for all the experimental
protocol due to its anatomical and physiological resemblance with the intestinal roundworm parasites of human beings.5, 6




ISSN: 2250-3013                                       www.iosrphr.org                                       395 | P a g e
IOSR Journal of Pharmacy
Vol. 2, Issue 3, May-June, 2012, pp.395-397


Anthelmintic screening: The anthelmintic activity was carried out as per the protocol of Ajaiyeoba et al.7 four groups of
approximately equal size Indian earthworms consisting of six earthworms in each group were used for the study. Each group
was treated as following:

     Group-I - Vehicle (DMSO in Distill water)
     Group-II - Piperazine citrate, Albendazole, Pyrantel pamoate (20 mg/ml)
     Group-III- Aqueous extract of leaves and pulp (20mg/ml)
     Group-IV- Ethanol extract of leaves and pulp (20mg/ml.)

    Observations were made for the time taken to paralysis and death of individual worms. Time for paralysis was noted
when no movement of any sort could be observed except when the worms were shaken vigorously. Death was concluded
when the worms lost their motility followed with fading away of their body colors.8, 9

Observation table:

                 Table 1: Anthelmintic Activity of ethanol extracts of Aegle marmelos against Earthworm
                                                                       Time taken to paralysis and death of worms
    Groups       Treatment                    Concentration
                                                                       Paralysis time (Min.)      Death Time (Min.)
                                              5% DMSO in Distill
    Control      Drugless                                              A                          A
                                              water

                 Aegle marmelos (Pulp)


                     a.   Aqueous                20mg/ml               32.38± 1.007                59.03± 0.3295

                     b.   Ethanolic              20mg/ml               33.75 ± 0.8167              54.16 ± 0.5757
    Test
                     Aegle            marmelos
                     (Leaves)

                     c.   Aqueous                20mg/ml               26.98± 0.7411               51.43± 1.182

                     d.   Ethanolic              20mg/ml               27.67±0.4875                55.07 ± 1.307
                 Albendazole                 20mg/ml                   28.76± 0.3812               50.39 ± 0.8105
    Standard     Piperazine Citrate          20mg/ml                   7.81±0.1661                 A
                 Pyrantel pamoate            20mg/ml                   21.60 ± 0.4647              64.58 ± 1.948
   Results are expressed as Mean± SEM, n=6 in each group
   A=absent of activity,

Result and Discussion
         In this anthelmintic study both the extracts (Aqueous and Ethanolic) of Aegle marmelos produce paralysis as well as
death of the worms. As shown in table2, aqueous and ethanolic extract exhibited anthelmintic activity as in standards (i.e.
Piperazine citrate, Albendazole and Pyrantel pamoate). Although, Piperazine citrate caused only paralysis not death of the
worms. Furthermore the anthelmintic effect of aqueous and ethanolic extract of Aegle marmelos was comparable with the
standard drugs, although it caused paralysis as well as death of worms similar to albendazole and Pyrantel pamoate.
Conclusion
          It can be concluded that the ethanolic and aqueous extract of Aegle marmelos leaves have profound anthelmintic
activity against tested worms but not better than standards. Further in future researches will also being carried out in our
department to establish the prescribed mechanism of action and to identify phytochemical moiety which is responsible for
this activity.

ACKNOWLEDGMENT


ISSN: 2250-3013                                      www.iosrphr.org                                      396 | P a g e
IOSR Journal of Pharmacy
Vol. 2, Issue 3, May-June, 2012, pp.395-397


        Authors are thankful to Prof .Pradeep Mishra Director, Institute of Pharmaceutical Research, GLA University for
providing the necessary facilities and support to carry out this work.

REFERENCE
      1.  Daniel Vivek, Daniela Kratika, Goyal Swapnil, Parihar S Mahendra and Singh Mangal. (2010). Synthesis and In-
          vitro Anthelmintic Activity of Some 4-aminophenol Derivatives. International Journal of Pharmaceutical
          Research, Vol 2, (2): 21-24.
      2. Chopra RN, Naver SC, ChopraIC. (1956).Glossary of Indian Medicinal Plants. New Delhi, CSIR 160.
      3. Hammond DA, Fielding D, Bishop SC. (1997). Prospects for plant anthelmintic in tropical veterinary medicine.
          Vet Res Comm, 21: 213-228.
      4. Kirtikar KR, Basu BD. (2005) .Indian Medicinal Plants .vol.2.International book distr., page no.
      5. Thorn GW,Admas RD,Braunwald E.Isselbacher KJ,Petersdorf RG.(1977).Harrisons Principles of internal
          Medicine, McGraw Hill Co, New York.
      6. Vigar Z. (1984).Atlas of medical parasitology, 2nd Edn. P.G.Publishing Housing, Singapore.
      7. Ajaiyeoba EO, Onocha PA and Olarenwaju OT. (2001).Invitro anthelmintic properties of buchholzia coriaceae
          and Gynandropsis gyanendra extract .pharm boil. 39:217-20
      8. Vagdevi HM, Latha KP, Vaidya VP, Vijayakumar M and Pai KS. (2001). Synthesis and Pharmacological
          screening of some novel naphtha (2, 1-b) Furo-pyrazolines, isoxazoles and isoxazolines, Indi. J. Pharma. Sci, 63:
          286-291.
      9. Boltan S:in pharmaceutical Statistic-practical and Clinical Applications.(1997).new York Marcel Dekker,67-78
      10. Thompson DP, Geary TG. (1995). The structure and function of helminths surfaces in Biochemistry and
          Molecular Biology of parasites 1st ed. Academic press, New York, pp.203-232.




ISSN: 2250-3013                                     www.iosrphr.org                                      397 | P a g e

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IOSRPHR(www.iosrphr.org) IOSR Journal of Pharmacy

  • 1. IOSR Journal of Pharmacy Vol. 2, Issue 3, May-June, 2012, pp.395-397 Comparative anthelmintic activity of Aegle marmelos Linn leaves and pulp Kishan Singh*, Krishn Kumar Agrawal, Jeetendra Kumar Gupta Institute of Pharmaceutical Research, GLA University Mathura, (U.P.)-281406 ABSTRACT In the present work, experiments were conducted to evaluate the possible anthelmintic effects of different extracts of Aegle marmelos leaves and pulp on Indian earthworm (Pheretima posthuma) at 20mg/ml concentration. Results were expressed in terms of time for paralysis and time for death of worms. Piperazine citrate, pyrantal pamoate and albendazole were used as a standards and DMSO in distill water as a control group. The result revealed that leaves extracts of Aegle marmelos has better activity that pulp extracts but not better than the standards. Keywords: Anthelmintic, Albendazole, Aegle marmelos, pyrantel pamoate INTRODUCTION Helminthes parasite infections are global problems with serious social and economic repercussions in the Third World countries. Parasitic helminths affect animals and man and animals than any single group of parasites, causing considerable hardship and stunted growth. Most diseases caused by helminthes are of a chronic, debilitating nature; they probably cause more morbidity and greater economic and social deprivation among humans The diseases affect the health status of a large fraction of the human population as well as animals.1 Herbal drugs have been in use since ancient times for the treatment of parasitic disease in human and could be of value in preventing the development of resistance.2, 3 A tree has unarmed branches and glabrous pedicles in pairs. The fruits are used as laxative and refrigerant. Prunes are often added to cathartic decoctions, improving their flavor and promoting their effect. They are also given in combination with other drugs in leucorrhoea and irregular menstruation. Literature survey revealed that plant extract has yet not been screened for anthelmintic activity. Therefore, the objective of this work was to assess the anthelmintic activity of Aegle marmelos leaves. Material and Methods Drugs and chemicals: Piperazine citrate, albendazole and pyrantal pamoate were used as standard drugs. Chemicals: Petroleum ether (60-80), Ethanol, Di methyl sulfoxide (DMSO) and sodium chloride were used in various part of experiment. Collection of plant material: Leaves of Aegle marmelos were collected from local area of Mathura and authenticated by Birbal Sahani Institute of Palaeobotony, Lucknow, Uttar Pradesh. (Voucher specimen no.13371/2011). Collected leaves were washed and dried at room temperature. After complete drying, leaves were powdered and passed through a sieve and stored in air tight container. Preparation of extracts: The Aegle marmelos were collected from Mathura (Uttar Pradesh) and dried in shade and coarsely powdered. It was than passed through the sieve no. 20. A weighted quantity (460g) of the powder drug was extracted with petroleum ether (60-800C) using soxhlet extractor. Defatted drug was subjected to ethanolic extraction and extract was dried by distilling off the solvent and then dried in desiccator. The marc collected after ethanolic extraction was subjected to aqueous extraction by maceration process for seven days consecutively and then extract was dried by evaporating the water and stored for further activity. Experimental worms: The earthworms of 3-5 cm in length and 0.1-0.2 cm in width were used for all the experimental protocol due to its anatomical and physiological resemblance with the intestinal roundworm parasites of human beings.5, 6 ISSN: 2250-3013 www.iosrphr.org 395 | P a g e
  • 2. IOSR Journal of Pharmacy Vol. 2, Issue 3, May-June, 2012, pp.395-397 Anthelmintic screening: The anthelmintic activity was carried out as per the protocol of Ajaiyeoba et al.7 four groups of approximately equal size Indian earthworms consisting of six earthworms in each group were used for the study. Each group was treated as following: Group-I - Vehicle (DMSO in Distill water) Group-II - Piperazine citrate, Albendazole, Pyrantel pamoate (20 mg/ml) Group-III- Aqueous extract of leaves and pulp (20mg/ml) Group-IV- Ethanol extract of leaves and pulp (20mg/ml.) Observations were made for the time taken to paralysis and death of individual worms. Time for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Death was concluded when the worms lost their motility followed with fading away of their body colors.8, 9 Observation table: Table 1: Anthelmintic Activity of ethanol extracts of Aegle marmelos against Earthworm Time taken to paralysis and death of worms Groups Treatment Concentration Paralysis time (Min.) Death Time (Min.) 5% DMSO in Distill Control Drugless A A water Aegle marmelos (Pulp) a. Aqueous 20mg/ml 32.38± 1.007 59.03± 0.3295 b. Ethanolic 20mg/ml 33.75 ± 0.8167 54.16 ± 0.5757 Test Aegle marmelos (Leaves) c. Aqueous 20mg/ml 26.98± 0.7411 51.43± 1.182 d. Ethanolic 20mg/ml 27.67±0.4875 55.07 ± 1.307 Albendazole 20mg/ml 28.76± 0.3812 50.39 ± 0.8105 Standard Piperazine Citrate 20mg/ml 7.81±0.1661 A Pyrantel pamoate 20mg/ml 21.60 ± 0.4647 64.58 ± 1.948 Results are expressed as Mean± SEM, n=6 in each group A=absent of activity, Result and Discussion In this anthelmintic study both the extracts (Aqueous and Ethanolic) of Aegle marmelos produce paralysis as well as death of the worms. As shown in table2, aqueous and ethanolic extract exhibited anthelmintic activity as in standards (i.e. Piperazine citrate, Albendazole and Pyrantel pamoate). Although, Piperazine citrate caused only paralysis not death of the worms. Furthermore the anthelmintic effect of aqueous and ethanolic extract of Aegle marmelos was comparable with the standard drugs, although it caused paralysis as well as death of worms similar to albendazole and Pyrantel pamoate. Conclusion It can be concluded that the ethanolic and aqueous extract of Aegle marmelos leaves have profound anthelmintic activity against tested worms but not better than standards. Further in future researches will also being carried out in our department to establish the prescribed mechanism of action and to identify phytochemical moiety which is responsible for this activity. ACKNOWLEDGMENT ISSN: 2250-3013 www.iosrphr.org 396 | P a g e
  • 3. IOSR Journal of Pharmacy Vol. 2, Issue 3, May-June, 2012, pp.395-397 Authors are thankful to Prof .Pradeep Mishra Director, Institute of Pharmaceutical Research, GLA University for providing the necessary facilities and support to carry out this work. REFERENCE 1. Daniel Vivek, Daniela Kratika, Goyal Swapnil, Parihar S Mahendra and Singh Mangal. (2010). Synthesis and In- vitro Anthelmintic Activity of Some 4-aminophenol Derivatives. International Journal of Pharmaceutical Research, Vol 2, (2): 21-24. 2. Chopra RN, Naver SC, ChopraIC. (1956).Glossary of Indian Medicinal Plants. New Delhi, CSIR 160. 3. Hammond DA, Fielding D, Bishop SC. (1997). Prospects for plant anthelmintic in tropical veterinary medicine. Vet Res Comm, 21: 213-228. 4. Kirtikar KR, Basu BD. (2005) .Indian Medicinal Plants .vol.2.International book distr., page no. 5. Thorn GW,Admas RD,Braunwald E.Isselbacher KJ,Petersdorf RG.(1977).Harrisons Principles of internal Medicine, McGraw Hill Co, New York. 6. Vigar Z. (1984).Atlas of medical parasitology, 2nd Edn. P.G.Publishing Housing, Singapore. 7. Ajaiyeoba EO, Onocha PA and Olarenwaju OT. (2001).Invitro anthelmintic properties of buchholzia coriaceae and Gynandropsis gyanendra extract .pharm boil. 39:217-20 8. Vagdevi HM, Latha KP, Vaidya VP, Vijayakumar M and Pai KS. (2001). Synthesis and Pharmacological screening of some novel naphtha (2, 1-b) Furo-pyrazolines, isoxazoles and isoxazolines, Indi. J. Pharma. Sci, 63: 286-291. 9. Boltan S:in pharmaceutical Statistic-practical and Clinical Applications.(1997).new York Marcel Dekker,67-78 10. Thompson DP, Geary TG. (1995). The structure and function of helminths surfaces in Biochemistry and Molecular Biology of parasites 1st ed. Academic press, New York, pp.203-232. ISSN: 2250-3013 www.iosrphr.org 397 | P a g e