In-Vitro Anthelmintic Activity on Aqueous and Ethanol Extracts of  Abies pindrow Bark

 

Shashi Pal Bharti1*, Hitesh Kumar1, Bharat Parashar1, Sandeep Deswal2, Nitika Chahar2 and Manish Devgan2

1Department of Pharmacy, Manav Bharti University, Solan, Himachal Pradesh, India

2Chemistry Research Lab., Faculty of Pharmacy, R.P. Educational Trust Group of Institutions, Bastara, Karnal, Haryana, India

*Corresponding Author E-mail:  palshashi31@gmail.com

 

ABSTRACT:

The anthelmintic activity of ethanol and aqueous extract of Abies pindrow was studied against earthworm Eisenia foetida (redworm). The ethanolic and aqueous extract was prepared by hot and cold respectively. Albendazole was used as the standard drug. The anthelmintic activity was carried out to study the effect of plant extract on adult earthworm Eisenia foetida (redworm), and noted down the paralysis time (Vermifuse), and death time (Vermicidal). It was found that both ethanolic and aqueous extract of bark of Abies pindrow found to have significant action against adult earthworm Eisenia foetida (redworm) at the dose of 25 mg/ml or more; which may be due to the presence of glycosides, carbohydrates, tannins and saponins in the extract. Thus it can be concluded that the bark of Abies pindrow can be used as anthelmintic agent.

 

KEYWORDS: Abies pindrow, Eisenia foetida and anthelmintic.

 

 

INTRODUCTION:

Herbal medicines is still the mainstay of about 75-80% of the world population, mainly in the developing countries, for primary health care because of better cultural acceptability, better compatiblibility with the human body and lesser side effects.1 Herbs have been used as food and for medicinal purposes for centuries. However, the use of medicinal herbs has increased over the past few years and research interest has focused on various herbs that possess hypolipidemic, antiplatelets, antitumour or immune stimulating properties that may be useful adjuncts in helping reduce the risk of cardiovascular disease and cancer.2 Helminthic infections are among the most widespread infections in humans, distressing a huge population of the world, although the majority of infections due to helminthes are generally restricted to tropical regions and cause enormous hazards to health and contribute to the prevalence of undernourishment, anemia, eosinophilia and pneumonia parasitic diseases cause ruthless morbidity affecting principally population in endemic areas. The gastro-intestinal helminthes becomes resistant to currently available anthelmintic drugs therefore there is a foremost problem in treatment of helminthes diseases.

 

Hence there is an increasing demand towards natural anthelmintics.3 Anthelmintics are those agents that expel parasitic worms (helminthes) from the body, by stunning gastro intestinal helminthes becomes resistant to currently available anthelmintic drugs therefore there is a foremost problem in treatment of helminthes diseases.4 A number of medicinal plants have been used to treat parasitic infections in man and animals.5-8 The literature survey reveals that use of plants as anthelmintic agent is still in infancy in modern system of medicine. Some of the plants which are reported as anthelmintic agent are Moringa oleifera, Vitex negundo9, Thespesia lampas (Cav.)10, Trigonella foenum-graecum11, Enicostemma littorale12, and Cassia tora13, Momordica charantia14, Thespesia lampas15, Capparis zeylanica16, Piper longum17, Cucurbita maxima18, Punica granatum19, Ficus racemosa20, Ficus benghalensis21. The present study aims to experimentally prove the Abies pindrow bark as anthelmintic.

 

MATERIALS AND METHODS:

Plant material

The bark of plant Abies pindrow was collected from forest of Kufari–Shimla (H.P.) in the month of March, 2013. The plant was taxonomically identified and authenticated as Abies pindrow by Dr. R. Raina, Professor and Head, Plant products, Herbarium and Museum Division, The herbarium sheet of the said sample is linked to UHF-Herbarium with field book no. 12423, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni-Solan (H.P.). The stem bark was dried at room temperature (30-40 C). The bark was pulverized in a mechanical grinder for prior to use.

 

Extraction

Bark of Abies pindrow was air dried, cut into small pieces and pulverized into a coarsely powdered drug. The powdered drug of the plant was subjected to hot extraction process in soxhlet apparatus with ethanol for 72 h and then macerated with distilled water for 8-10 days. The extracts were filtered and concentrated to dryness using a Lyophilizer. Dry extracts were stored at 4°C till further use. The ethanolic extract was reddish brown and aqueous extract was dark brown in color.

 

Experimental animals

Adult earthworm Eisenia foetida (red worm) were used for the evaluation of in vitro anthelmintic activity. They were collected from Agriculture Office and identified by Dr. J.L. Sharma, deputy Director, Solan (H.P.). The average length of earthworm was 2-6 cm and width was 0.1-0.3 cm.

 

Drugs and Chemicals

Albendazole (Glaxo Smithkline), ethanol (Jianghu Huaxi International Trade Co. Ltd., China).

 

Experimental Procedure

The anthelmintic activity was performed according to the method of Agrahari et al.22 The plant extract of Abies pindrow were evaluated for anthelmintic activity in Eisenia foetida (red worms) of nearly equal size (2-6 cm). Eisenia foetida is used to its anatomical and physiological resemblance with the intestinal roundworm parasite of human beings. Because of easy availability of earthworms, they have been used widely for the initial evaluation of the anthelmintic compounds. The worms were acclimatized to the laboratory condition before experimentation. The earthworms were divided into 10 groups of five earthworms in each and placed in petri dishes containing the extract solutions or the reference drugs as mentioned below-

 

Group-1: Received 1% gum acacia which is used as the control

Group-2: Received Albendazole suspension at a dose of 20mg/ml which served as the standard

Group-3: Received Ethanolic extract at a dose of 25mg/ml as 1% suspension of gum acacia

Group-4: Received Ethanolic extract at a dose of 50mg/ml as 1% suspension of gum acacia

Group-5: Received Ethanolic extract at a dose of 100mg/ml as 1% suspension of gum acacia

Group-6: Received Ethanolic extract at a dose of 150mg/ml as 1% suspension of gum acacia

Group-7: Received Aqueous extract at a dose of 25mg/ml as 1% suspension of gum acacia

Group-8: Received Aqueous extract at a dose of 50mg/ml as 1% suspension of gum acacia

Group-9: Received Aqueous extract at a dose of 100mg/ml as 1% suspension of gum acacia

Group-10: Received Aqueous extract at a dose of 150mg/ml as 1% suspension of gum acacia

 

All petri dishes were kept under room temperature. The living or viable worms kept under close observations. Observation was made for time taken to complete paralysis and death for individual worms. Each worm was frequently applied with external stimuli which stimulates and induce movement in earthworms, if alive. Paralysis was said to occur when the worms do not revive even in normal saline. Death was concluded when the worms lose their motility followed with fading of the body color. The motionless worms were transferred at 40°C to confirm that they were dead. The motionless worms were transferred at 400 C to confirm that they were dead.

 

RESULTS AND DISCUSSION:

Preliminary phytochemical screening of alcoholic extract revealed the presence of glycosides, saponins, carbohydrates, tannins and aqueous extract reveals the presence of glycosides, saponins, carbohydrates, tannins. Both ethanolic and aqueous extract of bark at conc. (25, 50, 100, 150 mg/ml) has shown a significant (P<0.01) anthelmintic activity when compared with the standard, i.e., albendazole (20 mg/ml), this can be clearly seen in the Table. The above discussed response is due to the presence of several phytoconstituents in the bark.

 

Table: Anthelmintic activity of ethanolic and aqueous extract of Abies pindrow bark.

Sr. No.

Group

Concentration

 (mg/ml)

Eisenia foetida (earthworm/redworm)

Time taken for Paralysis (P) in min.

(Mean and SEM)

Time taken for Death (D) in min. (Mean and SEM)

1

Aqueous extract

25

30.40 ± 15.53**

100.20 ± 2.28

50

17.40 ± 2.66**

64.00 ± 6.89**

100

17.00 ± 0.99**

39.60 ± 9.34**

150

06.60 ± 1.14**

19.20 ± 3.11**

2

Ethanolic extract

25

21.20 ± 4.76**

43.60 ± 4.27**

50

14.00 ± 6.81**

38.80 ± 4.20**

100

10.40 ± 2.88**

32.40 ± 6.10**

150

4.600 ± 2.40**

23.40 ± 4.77**

3

Albendazole

20

7.400 ± 2.79**

21.00 ± 6.20**

4

Control

--

--

--

**P<0.01 When compared with standard Albendazole

Results are mean ± SEM (n=5)

 

CONCLUSION:

From the above results it is concluded that alcoholic and aqueous extracts of bark of Abies pindrow have a potent anthelmintic activity which is comparable with standard drug albendazole. Further studies involving in-vivo model are required to find out and to establish effectiveness and pharmacological rationale for the use of bark of Abies pindrow as anthelmintic drug.

 

ACKNOWLEDGMENT:

The author is highly grateful to Dr. R. Raina, Professor and Head, University of Horticulture and Forestry- Nauni, Solan (H.P.) for identification of the plant; Dr. J. L. Sharma, Deputy Director, Agriculture Office, Solan for identification of earthworms and Department of Pharmacy, Manav Bharti University, Solan for providing facilities to carry out this work.

 

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Received on 31.03.2014       Modified on 18.04.2014

Accepted on 01.06.2014      ©A&V Publications All right reserved

Res.  J. Pharmacognosy & Phytochem. 6(3): July-Sept.2014; Page 112-114