IPSIndian Journal of Pharmacology
Home  IPS  Feedback Subscribe Top cited articles Login 
Users Online : 1025 
Small font sizeDefault font sizeIncrease font size
Navigate Here
  Search
 
  Next article
  Previous article 
  Table of Contents
  
Resource Links
   Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
   Article in PDF (296 KB)
   Citation Manager
   Access Statistics
   Reader Comments
   Email Alert *
   Add to My List *
* Registration required (free)

 
In This Article
   Abstract
   Materials and Me...
   Results
   Discussion
   Acknowledgment
   References
   Article Figures

 Article Access Statistics
    Viewed8008    
    Printed125    
    Emailed1    
    PDF Downloaded510    
    Comments [Add]    
    Cited by others 6    

Recommend this journal

 


 
RESEARCH ARTICLE
Year : 2008  |  Volume : 40  |  Issue : 2  |  Page : 64-68
 

Paralytic effect of alcoholic extract of Allium sativum and Piper longum on liver amphistome, Gigantocotyle explanatum


Division of Pharmacology and Toxicology, Indian Veterinary Research Institute, Izatnagar - 243 122, UP, India

Date of Submission20-Aug-2007
Date of Decision29-Apr-2008
Date of Acceptance05-May-2008

Correspondence Address:
D Kumar
Division of Pharmacology and Toxicology, Indian Veterinary Research Institute, Izatnagar - 243 122, UP
India
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0253-7613.41040

Rights and Permissions

  Abstract 

Objective: To investigate the effects of alcoholic extract of Allium sativum and Piper longum on the muscular activity of a parasitic amphistome, Gigantocotyle explanatum.
Materials and Methods: Amphistomes were isometrically mounted to record the spontaneous muscular activity by using Chart 4 software program (Power Lab, AD Instruments, Australia) and to examine the effects of cumulative doses (100, 300, 1000, and 3000 g/ml) of the plant extracts on the amplitude (g), frequency (per 10 min), and baseline tension (g) of the spontaneous muscular activity of the amphistome.
Results: Alcoholic extract of A. sativum produced significant reduction in the frequency and amplitude of contractile activity of the amphistome at 1000 and 3000 g/ml bath concentrations. Complete paralysis of the amphistome was observed after 15 min of addition of 3000 g/ml concentration. Alcoholic extract of P. longum also caused paralysis following 15-20 min exposure of the amphistome to 3000 g/ml concentration. In both the cases the amphistomes did not recover from paralysis following 2-3 washes.
Conclusion: The observations demonstrate the paralytic effect of alcoholic extract of A. sativum and P. longum on G. explanatum.


Keywords: Alcoholic extract, Allium sativum, Gigantocotyle explanatum, Piper longum, spontaneous muscular activity


How to cite this article:
Singh T U, Kumar D, Tandan S K. Paralytic effect of alcoholic extract of Allium sativum and Piper longum on liver amphistome, Gigantocotyle explanatum. Indian J Pharmacol 2008;40:64-8

How to cite this URL:
Singh T U, Kumar D, Tandan S K. Paralytic effect of alcoholic extract of Allium sativum and Piper longum on liver amphistome, Gigantocotyle explanatum. Indian J Pharmacol [serial online] 2008 [cited 2019 Oct 20];40:64-8. Available from: http://www.ijp-online.com/text.asp?2008/40/2/64/41040


Parasitic infection is a major health problem throughout the world and is responsible for considerable economic losses to the livestock industry, particularly to poor livestock owners in developing countries. Other adverse effects of these parasites include loss of meat, wool, and egg production. Amongst helminths, infection caused by trematodes like amphistomes ( Gigantocotyle explanatum and Gastrothylax crumenifer ) and Fasciola sp. is more serious than that due to round worms. The prevalence of G. explanatum is very high in ruminants in the Indian subcontinent. These parasites cause hemorrhages and connective tissue proliferation at the site of attachment, vacuolar degeneration in the liver, and hyperplasia in the bile duct, thereby seriously affecting the health and productivity of infected animals. [1]

Chemotherapy is the only efficient and effective tool to cure and control the helminth infection, as efficacious vaccines against helminths have not been developed so far. Indiscriminate use of synthetic anthelmintics in domestic animals has resulted in the development of resistance in helminth parasites. [2],[3],[4] Further, residual toxicity, adverse reactions, high cost, and inaccessibility to the rural farmers are problems associated with these agents. Consequently, there is an urgent need to develop newer, selective, and eco-friendly agents to control helminth infections. Plant-based anthelmintics offer an alternative to overcome some of these problems and they can be both sustainable and environmentally acceptable. Unlike synthetic anthelmintics, plant-based anthelmintics with different modes of action could be of value in preventing the development of resistance. [5] Herbal drugs have been in use since ancient times for the treatment of a variety of acute and chronic parasitic diseases, both in human and in veterinary medicine. [6],[7] The use of an extract of male fern ( Dryopteris felix mas ) against cestodes and trematodes and that of arecoline (from Areca catechu ) against tapeworms of dogs and poultry has also been reported. [8]

In the search for plant-based anthelmintics, extracts of different medicinal plants have been tested for action against flatworms and roundworms in vitro and in vivo and have been found to possess anthelmintic activity. For example, an alcoholic extract of Mallotus philippinensis caused complete paralysis of F. gigantica in vitro , [9] garlic protected mice against Schistosoma infection, A. sativum has shown anthelmintic action in vitro against Heterakis gallinae and Ascaridia galli , [10] Haemonchus contortus, [11] a free-living nematode of Rhabditis sp., larvae of Nippostrongylus brasiliensis , and eggs of Ascaris summ. [12] In vivo A. sativum has demonstrated activity against strongyloids in donkeys. [13] P. longum has been reported to produce paralysis of Ascaris lumbricoides. [14] Moreover, extracts of the plants used in the present study, A. sativum and P. longum , have shown good hepatoprotective activity in rats. [15],[16] Although A. sativum and P. longum have shown activity against roundworms, to the best of our knowledge, there is no study on the effects of these plants on G. explanatum . The present study evaluates the effects of alcoholic extract of A. sativum and P. longum on spontaneous muscular activity of G. explanatum .


  Materials and Methods Top


Collection of parasites

Mature and healthy G. explanatum were collected from the bile ducts of freshly slaughtered buffaloes from the local abattoir in warm (38 1C) Hank's balanced salt solution (HBSS) containing antibiotics (streptomycin sulphate - 6900 units @ 10mg/ml; and benzyl penicillin - 9900 units/liter). They were brought to the laboratory in an insulated container and were kept in the BOD incubator at 38 1C until further use.

Preparation of alcoholic extract of the plant material

The plant materials (dried fruits of P. longum and bulbs of A. sativum ) were pounded and then extracted with 70% ethanol under reflux. The alcoholic extract was concentrated under reduced pressure to a semisolid mass and made free from solvent.

Preparation of plant extract suspension

Suspensions (100 mg/ml) of the alcoholic extracts of A. sativum and P. longum were prepared in Tween-80 (final concentration 0.1%) and distilled water just before their use and further dilutions were prepared in HBSS solution.

Isometric mounting of G. explanatum and mechanical recording of the spontaneous muscular activity

Mature and active G. explanatum were mounted isometrically in HBSS solution at 38 1C as per the method described for Gastrothylax crumenifer. [17] Briefly, the amphistomes were mounted with the help of two fine steel hooks. One hook was inserted 1-2 mm caudal to the anterior sucker and fixed to the tip of an aeration tube and another hook was inserted through the surface of the acetabulum and connected to the transducer (Power Lab, AD Instruments, Australia).

The isometrically mounted parasite was equilibrated without any tension for 30 min, following which 300 mg tension was applied and spontaneous muscular activity was recorded using Chart 4 software program (Power Lab, AD Instruments, Australia). Control recordings were made for 15 min before the addition of a drug. During the equilibration period, the bath fluid was changed once every 10 min. Three parameters, namely, frequency (total number of contractions in 10 min), amplitude (average of all peaks per 10 min or average tension) of spontaneous muscular contractions, and baseline tension (average of all minimum levels of contractions used for measuring amplitude) of the isometrically mounted G. explanatum were measured. Measurements were made for a period of 10 min immediately before the application of a dose of the extract or before a wash following application of a dose.

Effect of cumulative concentrations of plant extracts on spontaneous muscular activity of isometrically mounted G. explanatum

Cumulative doses (100, 300, 1000, and 3000 g/ml) of the plant extract were added to the tissue bath with the isometrically mounted amphistome. Each dose was allowed to act for 15 min with concomitant recording of spontaneous muscular activity of the G. explanatum . The effects of various concentrations of plant extracts on frequency and amplitude of spontaneous muscular contractions and on the baseline tension of the isometrically mounted G. explanatum were recorded and compared with the control recording.

Statistical analysis

The results are presented as mean standard error of mean. To measure the level of significance, one way ANOVA with Tukey's multiple comparison tests were applied.


  Results Top


Recording of spontaneous muscular activity (SMA) of G. explanatum

The isometrically mounted G. explanatum exhibited SMA for several hours without significant change in amplitude, baseline tension, and frequency of the rhythmicity. The control (i.e., SMA recorded within 15 min of tension application) amplitude, baseline tension, and frequency of SMA were 0.31 0.03 g ( n = 6), 0.17 0.02 g ( n = 6), and 51.50 3.52/10 min ( n = 6), respectively. The amplitude (0.32 0.02 g; n = 6), baseline tension (0.18 0.02 g; n = 6), and frequency (51.83 1.33/10 min; n = 6) of spontaneous contractions recorded after a period of 2 h, were not significantly different from those recorded within 15 min after applying the tension to G. explanatum [Figure 1A],[Figure 1B].

Effect of cumulative concentrations of alcoholic extract of A. sativum on SMA of G. explanatum

The amplitude of contractions of the amphistomes was significantly decreased to 0.20 0.01 g ( P < 0.05) and 0.10 0.01 g, ( P < 0.001) with 1000 and 3000 g/ml concentrations of A. sativum extract, respectively, as compared to control (0.32 0.03). The extract, at 1000 and 3000 g/ml concentrations, also produced significant ( P < 0.01 and P < 0.001, respectively) reduction in the frequency of the SMA of amphistome. The values of frequency of SMA of amphistome at 1000 and 3000 g/ml concentration of the extract were 33.33 2.73 and 24.00 2.79, respectively. The alcoholic extract of A. sativum did not cause any significant change in the baseline tension up to a concentration of 1000 g/ml. However, at 3000 g/ml concentration, there was significant ( P < 0.05) reduction in the baseline tension of the SMA of the amphistome. The extract produced complete paralysis of the fluke after 15-20 min of administration of 3000 g/ml concentration [Figure 2A],[Figure 2B] and the SMA of the amphistome did not revive following 2 washes at 10 min intervals (data not shown). The recordings demonstrate a dose-dependent and progressive reduction in the SMA of the amphistomes from 100 to 3000 g/ml concentration of the extract.

Effect of cumulative concentrations of alcoholic extract of P. longum on the SMA of G. explanatum

In isometrically mounted G. explanatum , the normal amplitude, baseline tension, and frequency of contractions recorded were 0.33 0.02 g ( n = 6), 0.17 0.02 g ( n = 6), and 52.83 3.01/10 min ( n = 6), respectively. The average baseline tension was decreased significantly ( P < 0.01) with 3000 g/ml concentration of the extract of P. longum . The amplitude was reduced significantly to 0.19 0.01 g ( P < 0.001) and 0.11 0.02 g ( P < 0.001) at concentrations of 1000 g/ml and at 3000 g/ml, respectively, as compared to control. The frequency of SMA of the amphistome was significantly ( P < 0.001) reduced at 1000 g/ml and 3000 g/ml concentrations of the extract [Figure 3A],[Figure 3B]. Paralysis of the amphistome ensued after 15-20 min of administration of 3000 g/ml concentration of the alcoholic extract of P. longum . The paralyzed amphistome did not revive following 2-3 washes at 10 min intervals.


  Discussion Top


The main finding from this investigation was that alcoholic extracts of A. sativum and P. longum produced complete paralysis of G. explanatum after 15 min of addition of 3000 g/ml concentration of the extracts and the amphistomes did not recover from paralysis even after 2-3 washes.

Chemotherapeutic agents available for treatment of helminth infection act mainly through three different mechanisms, viz, by disruption of the neuromuscular physiology, by blocking the energy metabolism, and by disturbing the highly efficient reproductive system of the parasites. [18] Several important anthelmintics cause paralysis of helminth parasites by disrupting one or the other aspect of their neuromuscular system. [8],[19]

The muscular activity of helminth parasites can be appreciated as SMA, which can be recorded mechanically with the help of a physiograph. The SMA can be quantified in terms of frequency, amplitude of rhythmic contractions, and baseline tension and these parameters can be measured before and following drug treatment and the values compared. [20] The changes produced in the SMA of isometrically mounted amphistome by a drug, shows the involvement of the neuromuscular system on account of rapidity of action. Rapid and marked change in the SMA of an isometrically mounted parasite by a drug indicates that the neuromuscular system of the parasites has been affected. Thus, SMA of isometrically mounted parasite can be used to evaluate anthelmintic activity of new compounds in vitro . In the present study, the SMA of G. explanatum is grossly similar to that of G. crumenifer, [17] S. mansoni , [21] and F. hepatica. [22]

We have observed in a separate study that alcoholic extracts of A. sativum and P. longum inhibited the gross (visually assessed) motility of G. explanatum at 4 h of incubation (under publication). In the present study both the extracts induced marked decrease in the amplitude and frequency of rhythmic contractions at 1000 g/ml concentration and produced complete paralysis of the amphistome within 15 min recording in presence of their highest concentration (3000 g/ml). It can be appreciated from the recording of the effect of alcoholic extract of A. sativum that the inhibitory effect was dose-dependent from 100 to 3000 g/ml bath concentrations, although same could not be verified statistically. In case of P. longum also, a dose-dependent effect could be seen. Alcoholic extract of P. longum also demonstrated concentration-dependent inhibitory effect on the SMA of G. crumenifer from 1000 to 3000 g/ml bath concentrations. Thus, the paralysis of G. explanatum is the effect of the alcoholic extracts of A. sativum and P. longum .

The chemotherapeutic value of both the extracts is also evident from an earlier study, [23] wherein P. longum (fruits), along with Azadirachta indica (bark), Butea frondosa (seeds), and Nigella sativa (seeds), produced broad spectrum anthelmintic action against roundworms ( H. contortus and Oesophagostomum columbianum ) and flukes ( Paramphistomum cervi ) in calves. In vitro , P. longum produced paralysis of A. lumbricoides. [14] Furthermore, A. sativum has also been shown to possess anthelmintic action in vitro against H. gallinae and A. galli, [10] H. contortus, [11] etc., and in vivo against strongyloides in donkeys. [13] The present observations provide evidence for the paralytic effect of alcoholic extracts A. sativum and P. longum on amphistomes.

In conclusion, the observations demonstrate a paralytic effect of alcoholic extract of A. sativum and P. longum on G. explanatum by progressive reduction in the SMA, which may be associated with their inhibitory effect on the neuromuscular system of the amphistome.


  Acknowledgment Top


The authors are thankful to the Director, Indian Veterinary Research Institute, Izatnagar (U.P.), for providing the necessary facilities to complete this work.

 
  References Top

1.Swarup D, Pachauri SP, Mukherjee SC. Prevalence and clinico-pathology of naturally occurring fascioliasis and biliary amphistomiasis in buffaloes. Indian J Anim Sci 1987;57:252-56.  Back to cited text no. 1    
2.Singh D, Swarnkar CP, Khan FA. Anthelmintic resistance in gastrointestinal nematodes of livestock in India. J Vet Parasitol 2002;16:115-30.  Back to cited text no. 2    
3.Von Samson-Himmelstjerna G, Blackhall W. Will technology provide solutions for drug resistance in veterinary helminths? Vet Parasitol 2005;132:223-9.  Back to cited text no. 3    
4.Wolstenholme AJ, Fairweather I, Prichard R, von Samson-Himmelstjerna G, Sangster NC. Drug resistance in veterinary helminths. Trends Parasitol 2004;20:469-76.  Back to cited text no. 4    
5.Hammond JA, Fielding D, Bishop SC. Prospects for plant anthelmintics in tropical veterinary medicine. Vet Res Com 1997;21:213-28.  Back to cited text no. 5    
6.Chopra RN, Nayyar SL, Chopra IC. Glossary of Indian Medicinal Plants. New Delhi: Council of Scientific and Industrial Research; 1956. p. 160.  Back to cited text no. 6    
7.Kumar D, Rao GS, Raviprakash V, Tripathi HC, Tandan SK, Lal J, et al. Indigenous plants active against helminthic infections of domestic animals in India. Proc Natl Acad Sci India 2005;75:287-98.  Back to cited text no. 7    
8.Reinemeyer CR, Courtney CH. Chemotherapy of parasitic diseases. In: Adam, H.R. Editor, Veterinary Pharmacology and Therapeutics, 8 th Edition, Ames, Iowa State University Press; 2001.p.947-91.  Back to cited text no. 8    
9.Kushwaha DS, Kumar D, Tripathi HC, Tandan SK. Effect of some indigenous medicinal plant extracts on Fasciola gigantica in vitro. Indian J Anim Sci 2004;74:143-6.  Back to cited text no. 9    
10.Nagaich SS. Studies on the anthelmintic activity of Allium sativum (Garlic) oil on common poultry worms Ascaridia galli and Heterakis gallinae. J Parasitol App Anim Biol 2000;9:47-52.  Back to cited text no. 10    
11.Zafar-Iqbal, Nadeem QK, Khan MN, Akhtar MS, Waraich FN. In vitro anthelmintic activity of Allium sativum, Zingiber officinale, Cucurbita mexicana and Ficus religiosa. Int J Agric Biol 2001;3:454-7.  Back to cited text no. 11    
12.Chybowski J. Study of the anthelmintic activity of garlic extracts. Herba Polonica 1997;43:383-7.  Back to cited text no. 12    
13.Sutton GA, Haik R. Efficacy of garlic as an anthelmintic in donkeys. Israel J Vet Med 1999;54:23-7.  Back to cited text no. 13    
14.D'-Cruz JL, Nimbkark AY, Kokate CK. Evaluation of fruits of Piper longum and leaves of Adhatoda vasica nees for anthelmintic activity. Indian Drugs 1980;17:99-101.  Back to cited text no. 14    
15.El-Manakhy EM, Yaussef SA, Hassieb MM. The protective effect of garlic against carbon tetrachloride-Induced hepatotoxicosis in male rats: Some histopathologic and histochemical studies. Egyptian J Comp Path Clin Pathol 1998;11:35-45.  Back to cited text no. 15    
16.Rege N, Dhanukar S, Karandikar SM. Hepatoprotective effects of P. longum against carbon tetrachloride induced liver damage. Indian Drugs 1984;21:569.  Back to cited text no. 16    
17.Noble TG. In vitro studies on some medicinal plant extracts against Gastrothylax crumenifer, a rumen amphistome. M.V.Sc Thesis, Izatnagar: Indian Veterinary Research Institute; 2004. p. 21.  Back to cited text no. 17    
18.Geary TG, Klein RD, Vanover L, Bowman JW, Tompson DP. The nervous system of helminths as target for drugs. J Parasitol 1992;78:215-30.  Back to cited text no. 18    
19.Loukas A, Hotez PJ. Chemotherapy of helminth infections. In: Brunton LL, Lazo JS, Parker KL. Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 11 th ed. USA: McGraw-Hill Companies; 2005. p. 1073-93.  Back to cited text no. 19    
20.Sukhdeo SC, Sangster NC, Mettrick DF. Effects of cholinergic drugs on longitudinal muscle contraction of Fasciola hepatica. J Parasitol 1986;72:858-64.  Back to cited text no. 20    
21.Mellin TN, Busch RD, Wang CC, Kath G. Neuropharmacology of the parasitic trematode Schistosoma mansoni. Am J Trop Med Hyg 1983;32:83-93.  Back to cited text no. 21    
22.Fairweather I, Holmes SD, Threadgold LT. Fasciola hepatica: A technique for monitoring in vitro motility activity. Exp Parasitol 1983;56:369-80.  Back to cited text no. 22    
23.Raje AA, Jangde CR, Kolte SW. Evaluation of anthelmintic activity of mixture of indigenous medicinal plants in cow calves. J Vet Parasitol 2003;17:97-9.  Back to cited text no. 23    


    Figures

  [Figure 1A], [Figure 1B], [Figure 2A], [Figure 2B], [Figure 3A], [Figure 3B]

This article has been cited by
1 Study of antimicrobial activity and root symbionts of Hemionitis arifolia
Karmakar, J., Mukhopadhyay, S.K.
Physiology and Molecular Biology of Plants. 2011; 17(2): 199-202
[Pubmed]
2 Anthelmintic activity of Thirikadugu chooranam and its ingredients
Shree Devi, M.S., Sampath Kumar, B.
Asian Journal of Pharmaceutical and Clinical Research. 2011; 4(3): 58-60
[Pubmed]
3 Inhibitory effects of essential oil of allium sativum and piper longum on gross visual motility and glucose uptake of fasciola gigantica and gigantocotyle explanatum
Singh, T.U., Kumar, D., Tandon, S.K., Mishra, S.K.
Journal of Veterinary Parasitology. 2010; 24(1): 45-49
[Pubmed]
4 Inhibitory effect of essential oils of Allium sativum and Piper longum on spontaneous muscular activity of liver fluke, Fasciola gigantica
Thakur Uttam Singh,Dinesh Kumar,Surendra Kumar Tandan,Santosh Kumar Mishra
Experimental Parasitology. 2009; 123(4): 302
[Pubmed] | [DOI]
5 Inhibitory effect of essential oils of Allium sativum and Piper longum on spontaneous muscular activity of liver fluke, Fasciola gigantica
Singh, T.U., Kumar, D., Tandan, S.K., Mishra, S.K.
Experimental Parasitology. 2009; 123(4): 302-308
[Pubmed]
6 Anthelmintic activity of Trikatu churna and its ingredients
Reddy, B.U., Seetharam, Y.N.
Pharmacologyonline. 2009; 1(922): 927
[Pubmed]



 

Top
Print this article  Email this article

    

Site Map | Home | Contact Us | Feedback | Copyright and Disclaimer
Online since 20th July '04
Published by Wolters Kluwer - Medknow