INTERNATIONAL JOURNAL OF PHYTOTHEARPY RESEARCH Original Article Anthelmintic Activity of Moringa oleifera(lamk.)

INTERNATIONAL JOURNAL OF PHYTOTHEARPY RESEARCH ISSN 2278 – 5701

21

Original Article

Anthelmintic Activity of Moringa oleifera(lamk.)

Shrikant R. Inchulkar 1,2, B. Ravishankar1 , C.R.S. Pillai 1, J.M. Sharma1

1. Institute of Post Graduate Teaching and Research, Gujarat Ayurved University, Jamnagar, Gujarat, India

2. Government Ayurvedic College ,Raipur, CG, India

ABSTRACT

The quest for health and happiness is probably the most primitive of all quests. In Ayurveda there is some controversy as to the primitive formulation of a drug (1) Whether it was a single ingredients or (2) a compound of multiple ingredients. In veda, samhita and nighantu the use of single drug therapy for specific purpose as well as general purpose is prescribed. The prescription of Gana (dashemani ) falls under second purpose, in it the drugs may be used either as single or in compound formulation in any of the pharmaceutical preparations. Keeping this in view the drug Akshiva seeds(Shobhanjana- Moringa oleifera) the constituents of krimighna Dashemani has been evaluated for its krimighna effect (Anthelmentic Activity ) experimentally to the claims made in the classics, as krimi roga is most commonly occurring diseases in our country.During experimental trails its anthelmintic activity was evaluated in different species taking at least one representative each from cestodes, trematodes and nematodes ( in vitro condition), in two different preparation. Simultaneously the root bark of drug was also evaluated to find out whether it could be substituted for seeds, when seeds as not available at the same time to know the probable mechanism of action of drug preparations, studies were also carried out on isolated tissue preparation (i.e. rabbit jejunum and worms) as well as on faecal output and stool characteristics in mice.

Keywords: Moringa oleifera , Krimi roga , Anthelmintic activity, invitro studies.

INTRODUCTION

Vedas are considered to be the oldest among the existing religious expositions. They have a rich documentation of contemporary social, cultural and economic mores. It is not surprising that all the encompassing Vedas strolled into the

medicinal field too (Sharma DC). Shobhanjana too finds its description in the Vedas. In the Rigveda it is mentioned as Shigru for Janpad Vishesh (Rigveda 1951). Atharva parishista prescribing it for Ucchaatana Karma ( Atharvaveda, 1987).


22

Kaushik sutra prescribes its use along with butter in krimiroga (Kaushik sutra 1972). Several Dharma sutras forbid its use in washing and cleaning of teeth as well as in funeral rites (Dravya guna vigyana, samvat 2038). Thus it is clear that shobhanjana was widely used right from the vedic period.

Krimi Roga is a most commonly occurring disease, due to prevailing conditions of proverty, poor hygienic conditions and poor nutritional status in our country. (S.R.Inchulkar, M.S. Baghel, AYU, 1991). In the present study, Shobhanjana Beeja (Akshiva seeds) a constituent of the Krimighna Dashemani has been evaluated for its Krimighna effect experimentally. This is being done to provide an experimental basis to the claims made in the classics. During experimental trials its anthelmintic activity was evaluated in different species taking at least one representative each from cestodes, trematodes and nematodes (in invitro conditions), in two different preparations. Simultaneously, the root bark of the drug was also evaluated to find out whether it could be substituted for seeds, when the seeds are not available. At the same time to know the probable mechanism of action of the drug preparation, studies were also carried out on isolated tissue preparations (i.e. Rabbit jejunum and worms) as well as on faecal output and stool characteristic in mice. MATERIAL AND METHODS Preparation of extracts:

1. The Total Methanolic Extract of Seeds (TME): It was prepared from pharmacognistically identified plant by continuous process of extraction with a soxhlet apparatus and was dried over a hot

water and used for experimentation in the form of 3% suspension in Tween 80.

2. Cold Aqueous Infusion of Root Bark (CAI): 25 g. of coarsely powdered root bark from pharmacognostically identified plant material was poured into a conical flask containing 100 ml of distilled water and periodically mixed with thorough shaking and kept at room temperature for 48 hours. The solution was filtered using an ordinary filter paper and the filtrate was stored in a refrigerator till its usage for experimental studies.

3. Kwatha of Seeds (KW): This was prepared using standard procedure. 25 g of coarsely powdered (between mesh No. 40 - 60) seed material was mixed with 200 ml of tap water and the solution was reduced to 1/8 of its original volume by warming over a hot plate. The resultant kwatha was filtered and kept in a refrigerator till its usage for experimentation. Part I- Invitro studies Worms for the Study: The helminthes were collected at the slaughter houses from the intestine of goats, sheep , hens and from the liver of sheep and within half an hour of their slaughter they were transported to the laboratory in a thermos flask containing tyrode solution. The worms were thoroughly washed, cleaned and kept in tyrode solution at 34 ± 1°C. The worms were identified by examining morphological and anatomical features. The helminthes used in the study were Ascaridia galli, Taenia Saginata, Hymenolepis nana and fasciolepsis hepatica. The worms selected for experimental purpose were tape worms with scolax intact


23

and adult round worms. Only those worms showing vigorous spontaneous activity were used for experimentation. To determine viability of affected worms, pinch technique ( Holten and hot water technique (Garg et al) were used. The extracts were evaluated for anthelmintic activity in invitro experimental set-up. Petri dishes (4” Diameter) containing 25 ml of tyrode solution were taken - control dish containing 3% Tween 80 solution, and test extracts in different concentrations, which are given below were added to the dishes before placing worms. Either 5 tape or round worms or when small 6 to 7 or 6 liver flukes were placed in each dish. After placing the worms, the dishes were transferred to an incubator and incubate at 340C±10C. The worms were observed for spontaneous movement every hour, up to 4 to 8 hours and then once in 24 hours, if they were found to be without movement they were further tested by pinch and hot water method to ascertain in whether they are alive or dead.The gradation of mortality was done as follows: Motile worms were given the code “A“. If the worms were almost immobile but if after keen observation some movement were noted they were termed as sluggish “S” or very sluggish “VS’ and when even after pinch and hot water technique if no motility was noticed , worms were noted as dead “D;. Dose

A. For tape worms: - T. Saginta 1. TME of the seeds – 2 and 3 mg/ml of the solution. 2. CAI of the root barks – 8.25, 16.50, and 50 mg/ml of the solution. 3. KW of the seeds – 500, 1000 and 2000 mg/ml of the solution. H.nana 1. TME of the seeds – 1.25, 2 and 2.5 mg/ml of the solution. 2. CAI of the root bark – 16.25 and 50 mg/ml of the solution. 3. KW of the seeds – 500 and 1000 mg/ml of the solution. B. For liver flukes : - F. hepatica 1. TME of the seeds – 1 and 3 mg/ml of the solution 2. CAI of the root bark – 10 and 30 mg/ml of the solution. C. For round worms: - A. galli 1. TME of the seeds – 2 and 8 mg/ml of the solution. 2. CAI of the root bark – 8 and15 mg/ml of the solution. 3. KW of the seeds – 500 and 1000 mg/ml of the solution.

Part II - Studies on Isolated Tissue Preparations 1. Rabbit Jejunum : Young rabbit fasted over night was sacrificed by sharp blow on the head and exsanguinations of carotid arteries. The abdomen was opened by a middle line incision. Jejunum was identified and a piece of suitable length was excised

out, cleaned of extraneous connective tissue and mounted in an isolated organ bath containing aerated ringer’s solution. Pendular movements of the tissue were recorded on a kymograph with an isotonic lever system (1:10 magnification with 1 g tension). Initially dose response curve with acetylcholine were recorded to choose sub


24

maximal dose for further studies. The effect TME perse, CAI perse and KW perse and modifying effects if any, on acetylcholine induced contractions were noted. 2. Worms: As mentioned earlier the collected worms, showing vigorous spontaneous activity were used for this experimentation. A piece of suitable length of each worm was excised out and mounted in an isolated organs baths containing tyrode solution. Rhythmic movement of each worms were recorded on a kymograph with isotonic lever with frontal writing points (1:7 magnification with 1 g. tension). The effect of TME, CAI and KW were recorded on T. sangita and A. galli. PART III - Effect of Shobhanjana Preparation on Faecal Output and Stool Characteristic in Mice TME of seeds, CAI of root bark and KW of the seeds of the Shobhanjan were studied for their effect on faecal on faecal out put and stools formation in mice. Albino mice (Swiss-albino) of either sex weighing between 15-32 g. were allotted to 4 different groups, to the first group distilled water was administered and served as control. Second group received TME in the dose of 1000 mg.kg-1, to the third group CAI of root bark was administered in the dose of 5000 mg. kg-1 and fourth group was given Kwatha of the seeds in the dose of 10 ml kg-1. The drug preparations were administered by gavage daily for 7 consecutive days. After drug administration the mice in individual

groups were transferred to metabolism cages and stools were collected in a paper, placed in the separator of the metabolism cages. Stools were periodically observed every day for changes in the consistency and characteristics. 24 hour faecal outputs were collected and dried by placing them in a incubator for 6 hours of 500C and weighted in a single pan balance. RESULTS PART I Invitro Studies Effect on Tape Worm:- T.saginata TME in the dose of 2 & 3 mg/ml. of physiological salt solution (tyrode) did not affect the worms.

‐ CAI of root bark in the lower doses ( 8.25 & 16.5mg/ml ) had no effect , but 2/5 worms kept in 50 mg/ml of tyrode solution became sluggish one hour after their placing in the tyrods solution and remained like that till their death.

‐ KW in the dose of 500 mg/ml. killed 1/6 worms at 5th hours of incubation, 2 became sluggish. 3 were dead and 3 became sluggish at 6th hour. At the dose of 1000 mg/ml, 2 worms died and 2 worms became sluggish at 5th hours and by 6th hours 3 were dead and 3 became sluggish. At still higher dose (2000 mg/ml) 4 worms died and 2 became sluggish at 5th hours, and at 6th hour 5 were dead and the remaining one became sluggish.( Table-1)


25

Cold Aqueous Infusion I Control I Mebendazole II

Cold Aqueous Infusion II Cold Aqueous Infusion III Mebendazole I

Control II

Fig-1. Effect of Moringa Oleifra on Tape Worms (T.saginata), in invitro condition.


26

Table No. – 1: Effect of shobhanjana preparations on tape worms (T.saginata) in invitro conditions.

Name of the preparation in mg/ml.

Observation made at different time Intervals (hrs.)

0 1 3 5 6 24 Control A= 5 A= 5 A= 5 A=5 A=5 D = 5

TME - 2 - 3

A= 5 A= 5

A= 5 A= 5

A= 5 A = 5

A= 5 A= 5

A= 5 A = 5

D = 5 D = 5

CAI - 8.25 - 16.50 - 50

A = 5 A = 5 A = 5

A= 5 A= 5 S = 2 A = 3

A= 5 A = 5 S = 2 A = 3

A= 5 A= 5 S = 2 A = 3

A= 5 A = 5 S = 2 A = 3

D = 5 D = 5 D = 5

Control A= 6 A = 6 A = 6 A = 6 A = 6 KW -500

A= 6

A= 6

A=6 A=6

D = 3 S = 2 A= 1

-1000 A= 6 A= 6

S = 1 A = 5

D = 2 S = 2 A = 2

D = 3 S = 3

-2000 A = 6 A = 6 S = 2 A = 4

D = 5 S = 1

D = 4 S = 2

Key to abbreviations:

A = Active, S = Sluggish, VS = Very sluggish, D = Dead, TME = Total Methanol Extract of seeds, CAI = Cold aqueous Infusion of root bark, KW = Kwatha of seeds.


27

SME II Cold Aqueous Infusion I Cold Aqueous Infusion III

SME I Mebendazole Control

Control II

Fig 2. Effect of Moringa Oleifra on Tape Worms (H.nana), in invitro conditions.

H.nana : - In one set of experiment TME in the dose of 2 mg/ml of tyrode solution caused death of all worms (7/7) after 24

hours compared to worms in control dish which lived for more than 3 days.

In the second set at the dose of 1.25 mg/ml, 4/6 worms became sluggish after 8 hours. At


28

higher dose of 2.5 mg/ml of tyrode solution 6/6 worms became sluggish at 2nd hour after incubation and most of them dead by 8th hours compare to control where all the worms were active even after 24 hours. CAI of root bark of the lower dose (16.25 mg/ml)

had only weak effect at higher dose level (50 mg/ml) worms became sluggish after 24 hours. KW of the seed powder in the doses studied (500 and 1000mg/ml) did not produce any effect on H.nana tape worms (Table No. – 2 ).

Table No. – 2: Effect of shobhanjana preparations on tape worms (H.nana ) in invitro conditions.

Name of the preparation in mg/ml.


0 2 4 6 8 24 3rd day Control A= 7 A= 7 A= 7 A= 7 A= 7 A= 7 A= 7

D= 7 TME - 2 Control

A= 7 A= 6

A= 7 A= 6

A= 7 A= 6

A= 7 A= 6

A= 7 A= 6

A= 7 A= 6

TME -1.25 - 2.5

A= 6

A= 6

A= 6

S= 6

S = 1 A= 5 S= 6

S = 1 A= 5 S= 6

S = 4 A= 2 D= 5 S = 1

S = 4 A= 2 D= 6

CAI -16.25 - 50

A= 6

A= 6

A= 6

A= 6

A= 6

A= 6

A= 6

A= 6

A= 6

S = 2 A = 4

S = 3 A = 3 S = 6

Control A= 6 A = 6 A = 6 A = 6 A = 6 D = 6 KW -500 A= 5 A= 5 A= 5 A= 5 A= 5 D = 5 -1000 A= 5 A= 5 A= 5 A = 5 A = 5 D = 5

In the experiments in one set mebendazole was used in the form of 3% Tween 80 suspension and it did not affect the worms in the dose of 5mg/ml. of tyrode solution . After the reference given in the IPC, the tablets were dissolved in few drops of formic acid and diluted to requisite concentration with tyrode soln. With this the worms were dead within 10 min. of addition to the dishes. However, few drops of formic acid added to the 2nd vehicles control groups also caused death of the worms indicating that the death in the previous groups was perhaps due to the solvent. Hence, the result

could not be effectively compare with a relative standard. However in the experimental conditions extracts were found to be effective.

Effect on Liver Flukes :F.hepatica

TME: In the Petri dishes containing 1 mg/ml of extract in tyrode solution, 3/6 flukes became sluggish at 4th hour after incubation. With the higher dose (3 mg/ml) 3/6 were dead at 4th hour of incubation.

CAI of root bark at the same dose of 10 mg/ml of tyrode solution caused death of 3/6


29

flukes and with higher dose (30 mg/ml) all 6/6 flukes were dead at 4th hour of incubation.

Mebendazole in the form of suspension in 3% Tween 80 solution at the dose of 2 mg/ml caused death 5/6 flukes at 4th hours, of incubation (Table -3 ).

Seeds (TME) 1mg/ml Root bark (CAI) 10mg/ml Mebendazole 2mg/ml

Seeds (TME) 3mg/ml Root bark (CAI) 30mg/ml Control (Tyrode)

Liver Flukes

Fig- 3. Effect of Moringa Oleifra on liver flukes (F. hepatica), in invitro conditions.

Table-3: Effect of Shobhanjana preparations on liver flukes (F. hepatica) in invitro condition.

Name of preparation in mg/ml.

Observation made at different time Intervals (h A=6rs.)

0 1 2 3 4 Control A = 6 A = 6 A = 6 A = 6 A = 6 TME – 1 A = 6 A = 6 A = 6 S = 2 S = 3 A = 4 A = 3 – 3 A = 6 A = 6 S = 2 S = 2 D = 3 A = 4 A = 4 S = 3 CAI – 10 A = 6 A = 6 S = 2 S = 3 D = 3 A = 4 A = 3 S = 3


30

Kwatha CAI Root Bark 8mg/ml

Standard TME Seeds 4mg/ml Control

Fig 4a : Effect of M. Oleifera on round worm (A. galli ), in invitro conditions.

Effect on Round Worms: A. galli TME in the dose of 2 mg/ml. of incubation medium , made the worm ( 6/6 ) very sluggish at 24th hr. of incubation –compared to control where only 1/6 became very sluggish at that time. At the higher dose level of 8 mg/ml. 3/6 worms became

sluggish at 2nd hr., 4/6 at 3rd hr.,6/6 at 4th hr. and 1/6 found dead at 6th hr. and the remaining became very sluggish. CAI of root bark treated worms were not affected significantly at the dose levels studied i.e.8 and 15 mg/ml. of incubation medium.

– 30 A = 6 A = 6 S = 2 S = 3 D = 6 A = 4 A = 3 Mebedazole -2 A = 6 A = 6 S = 2 S = 4 D = 5 A= 4 A = 2 S = 1


31

Table-4: Effect of Shobhanjana preparation on round worms (A.galli) in invitro conditions.

Name of the preparation in

mg/ml.


0 1 2 3 4 5 6 24 48 Control

A=6 A= 6 A=6 A= 6 A=6 A=6 S=2

A=4

VS=1

A=5

Vs=1

A=5

TME - 2 A= 6 A= 6 S = 1 A = 5

S=1 A=5

S=1 A=5

S=1 A=5

VS=1 S=1 A=4

VS=6 Vs=6

- 8 A= 6 A= 6 S=3 A=3

S=4 A=2

S=6 VS=6 D=1 VS=5

D=2 VS=4

D=4 VS=2

CAI - 8 A= 6

A= 6

A=6

S=1 A=5

S=2 A=4

S=2 A=4

VS=2 S=2 A=2

D=1 VS=5

D=1 VS=4

-15

A= 6

A = 6

S=1 A=5

S=1 A=5

S=1 A=5

S=2 A=4

VS=1 S=2 A=3

VS=6

VS=6

KW -500 A= 6 A= 6 A=6 A=6

A=6

S=3 A=3

D=1 S=5

D=1 VS=5

D=2 VS=4

-1000 A= 6 A= 6

A=6

S=3 A=3

S=4 A=2

D=1 VS=5

D=3 VS=3

D=3 VS=3

D=5 VS=1

Piperazine -10 A=7 A=7 S=3 A=4

S=4 A=3

S=4 A=3

S=4 A=3

S=6 A=1

VS=6 A=1

D=7

KW of seeds at the dose of 500 mg/ml., made 3/6 worms sluggish at 5th hr. of incubation. One worm was dead and 5 became sluggish at 6th hr. and 24th hr. two dead and 4 very sluggish at 48th hr. at the dose level of 1000 mg/ml.,3/6 were sluggish

at 3rd hr of incubation .4 became sluggish at 4th hr. One worm was found dead and remaining 5 were very sluggish at 5th hr. By 6th ht. 3/6 were dead and 3 became very sluggish. The same was noted at 24th hr.


32

In the piperazine treated group (10mg/ml of the incubation medium ), 3/7 worms became sluggish at 2nd hr. of incubation ,4/7 at 3rd and 4th hr. all the 6 became sluggish at 6th hr.

and very sluggish by 24th hr ( this group served as relative standard for comparison of the drug effects) Table – 4.

PART II – Studies on isolated tissue preparations

(1) Rabbit jejunum: TME Kwatha of seeds and CAI of root bark had no effect perse. However, CAI in the dose level of (10mg/ml.) produced complete antagonism of ACh induced contraction. TME in the dose of 500 µg and 1mg/ml/ produced marked antagonism of ACh induced contractile response .Kwath in the doses studied (50 and 100mg/ml) had no effect on the tissue responses to Ach (Fig -5).

(2) Worms-

(A) Tape worms –T. Saginate.

TME of seeds at the dose of 500µg and 1000 µg/ml of bath fluid produced weak contraction. After washing out the drug, worm did not regain rhythmicity (Fig-6a). CAI of root bark at the lower dose (5mg/ml of bath fluid) produced contractile response. However, at higher dose level (10 mg/ml the contractile response was not noted (Fig-6a). KW of seeds in the dose studied ( 0.2 ml/40ml baths equivalent to 5 mg/ml of seed material) did not affect rhythmic movements of worms(Fig -6b).

Fig -5. Effects of M.Oleifera extracts on Rabbit jejunum.

S : Submaximul response to acetylcholine (ACh) 1 um /ml of bathfluid.

KW : Response to ACh in presence of seed kwatha; 0.1 ml /40ml of bathfluid .


33

A & A1 : Response to ACh in presence of CAI; 5 & 10 mg/ ml of bathfluid .

B & B1: Response to Ach in presence of TME; 500 & 1000ug/ mg/ ml of bathfluid.

Fig 6a and Fig 6b

Effects of M.Oleifera extracts on tape worm (T.Saginata).

C : Control Response.

A & A1 : Response to CAI : 5 & 10 mg /ml of bathfluid respectively.

B & B1 : Response to TME : 500 ug& 1 mg /ml of bathfluid respectively.

W : Change in the bathfluid.

KW & KW1 : Response to seed Kwatha ; 0.2 & 0.4 ml/40ml of bathfluid

( equi. to 50 & 100 mg of seed powder/ml of bathfluid)

(B) Round worms – A galli -TME of seeds at both the dose level studied

(500ug/ml and 1000ug/ml of bath fluid), the extracts did not produced any marked effect. Only slight increase in the amplitude of rhythmic contraction was noted (Fig-7b).

-CAI of root bark at both the dose level (5 and 10 mg/ml of bath fluid), a moderate contractile response was noted. However, after the higher dose rhythmicity was

completely abolished which reappeared after repeated washing (Fig-7a).

-KW of seed at the dose level studies (0.2 and 0.4 ml/40ml bath fluid equivalent to 5 and 10 mg/ml of seed material), did not produced any significant effect on the rhythemicity of the worm (Fig-7b).


34

Legends to the figure:

Fig 7a and fig 7b

Effects of M.oleifera extracts on Round worms (A .galli)

C : Control Response

A & A1 : Responses to CAI : 5 & 10 mg /ml of bathfluid respectively.

B & B1 : Responses to TME : 500 ug& 1 mg /ml of bathfluid respectively.

W : Change in the bathfluid.

KW & KW1 : Responses to seed Kwatha ; 0.2 & 0.4 ml/40ml of bathfluid ( equi. to 50 & 100 mg of seed powder/ml of bathfluid)

PART III-

Effect of shobhanjana preparation on faecal output and stool characteristics in mice.

None of the drug preparations administered could produce changes in the consistency

and characteristics of stools. Which looked normal and same in all the 4 groups. However, faecal output per day significantly decreased in drug treated groups compared to control as could be seen from the data presented in (Table 5).


35

Table 5. Effect of shobhanjana preparation on 24 hrs faecal output in mice.

Treatment (mg.kg-1)

Twenty four hour faecal output (g)

Mean SEM

‘P’ value

Control (Distilled water)

5.79 ± 0.11 -

TME(1000) 4.45 ± 0.28 < 0.01 CAI (500) 4.28 ± 0.13 < 0.01 KW (10 ml.kg-1 equivalent to 10 g. kg-1 of seed material)

4.68 ± 0.21 < 0.01

DISCUSSION

Parasitic worms infesting human belong to a wide spectrum of zoological species and vary with regard to their morphology, physiology, drugs sensitivity and host habitat. Hence, it will be very much difficult to find out the exact mechanism of anthelmintic activity found in any new drug. Before attempting to discuss the mode of anthelmintic activity found in Shobhanjana preparations, it would be useful to briefly dwell on the mechanism of anthelmintic activity of some of the well known drugs.

The best known mechanism of action is depolarizing neuromuiscular blocking as represented by pyrental pamoate and piprazine which cause paralysis of the worm muscle resulting in expulsion of the worm by peristalsis. However, there is different in the mechanism of two drugs. Whereas pyrental pamoate causes depolarization and increase spike discharge frequency accompanied by increase in tension in single

muscle cells of helminthes. Piprazine cause hyperpolarisation with reduction in spike discharge frequency in them. Pyrental also inhibits the activity of the enzyme acetylcholine esterate further potentiating its effects on neuromuscular junction. The second well known mechanism of action is inhibition of oxygen and glucose uptake as exemplified by pyruvium pamoate. Hycanthone like drugs interfere with the laying of eggs, induces separation of paired worms and degenerative changes. The mechanism of action on this is assumed to be due to the stimulation of serotonin uptake into the non neuronal tissue, depriving neuronal tissue of its putative excitatory neurotransmitter.

Thiabendazole like drugs prevent embryonic development of ascaris eggs invitro. Niridazole produces vermicidal activity by interfering with activity of reproductive system of the worm. Tetrachloroethalene types of drugs are presumed to cause death of helminthes by interfering with the release


36

of lysosomal enzymes which are essential for the intracellular digestion of absorbed nutrients.

The main focus of our study was to evaluate Shobhanjana prepration for anthelmintic activity to provide an experimental basis to the claims made in the classics. Perusals of the result and of the study reveal that two of the preparation possesses significant anthelmintic acitivity i.e. KW and TME of seeds.

Of the three preaparation studied – Total methanolic extract (TME) of the seed possess significant anthelmintic activity against tape worms of H.nana variety , however its effect was not significant against T.saginata – pointing towards the necessity of taking into consideration the change of sensitivity due to species differences. It has moderate effect against liver flukes. and against round worms TME possess marked paralyzing effect which compares quite favorably with the activity obtained in the relative standard i.e. Piperazine. Presence of significant anthelmintic activity in TME is a pointer towards the necessity of elucidating the mechanism of action and isolation of active principle(s), responsible for the activity, from the extract.

Cold aqueous infusion (CAI) of the root bark also possesses significant anthemintic activity against liver fluke. However its activity against tape worms was not marked and it seems to be ineffective against rounds worms. Considering the easiness of its

preparation, it could play an important role in therapy of fluke infestation provided it is found effective in invivo conditions.

Kwatha of the seeds (KW) –unlike TME of seed possess significant activity against T. saginata variety while it has no effect on H.nana. Like TME it also possesses significant vermicidal activity against round worm (A.galli). Easiness of preparation makes it preferable to TME – which cannot be prepared readily at home. Since it is equally effective as TME it would be interesting to isolate and characterize active principle in the preparations. The active principle seems to be heat stable. Further evaluation in invitro conditions is necessary to give unequivocal basis to the recommendation in the classics. Since CAI of root bark is effective only against liver flukes, it cannot be used as substitute to seeds in tape worm and round worm infestations.

It can be used in liver fluke infestation as a substituted to seed preparation if need arises While coming to the probable mechanism of action the activity of the extracts on rabbit jejunum do not provide clear evidence – while CAI and TME produced acetylcholine antagonism, kwatha had no effect, but in anthelmintic activity evaluation, kwatha and TME had significant activity against round worms while CAI had no effect.

In isolated worm’s experiments TME produced weak contraction of T.saginata and significantly decreased the rhythmicity and on round worms it had no marked


37

effect, indicating that worm muscle contraction do not add significantly to the anthelmintic activity of the extracts. Kwatha of the seeds also did not produced any marked effect on round worm, again indicating worms muscle paralysis is not the mechanism of action of the preparation. Since none of the preparation could affect the consistency and characteristics of stools in mice, involvement of hypermotility of the G.I tract of the host a possible contributing factor to the probable anthelmintic activity is ruled out. The decrease in the 24 hr faecal out put may be due to some other reason.

Result of the study indicate that Shobhanjana preparation do possess

anthelmintic activitry, though narrow in the spectrum, of worms affected. It would be interesting to elucidate the probable mechanism of activity by studying on the parameters reprentative of the mechanisms enumerated in the beginning of this discussion.

Though experimental studies were also planned to study the effect of shobhanjana preparation on larva of Anchylostomaduodenale and Necator americunnus, but they could not be completed due to technical difficulties in separationg the developed larvae.

REFERENCES

1. Sharma D.S. “Vedon me dravya guna shastra: M.D. (Ay) Thesis, G.A.U.,Jamnagar.

2. Rigveda with Sayan Bhashya; Edi.1st, Pub. By sontakke,N.S., vedic sanshodhan mandal,poona ,1951.

3. Atharvaveda with Sayan Bhashya & Hindi trans.by Pd. Ramachandra Sharma, Edi 1st, Pub. Sanatan Dharma Press, Moradabad,1987.

4. Kaushik sutra by Bloomfield, M.; Edi 1st, Pub. Motilal Banarasidas 1972.

5. Dravya guna vigyan Part IV, V; by Sharma, P.V., Edi. 1st, Pub. Chowkhambha Bhartiya Academy, Varanasi, Samvat 2038.

6. S.Inchulkar, M S Baghel “ Correlation of krimis classifications and terminologies with the parasites of Modern Medicines (An retrospect) ,AYU, 1991, 1-12.

7. Holton (1947) Quoted from shrivastava et al “Anthelmintic activity of Cucurbita maxima (kaddu) seeds.” Ind.Jour. Med. Res., 55, 6, June, 1967.

8. Garg, L.C. & atal , C.K. “ Anthemintic activity of calotropain & bromelain “; Ind. Jour.Pharm. 25,422,1963.

9. Goodman & gilman “The pharmacological basis of therapeutics “Edi. 5th, Pub. Macmilan Publishing Co. INC, New York, 1975, pp. 1018-1042.

Documents

INTERNATIONAL JOURNAL OF PHYTOTHEARPY RESEARCH Original Article Anthelmintic Activity of Moringa oleifera(lamk.)