Anthelmintic and Antibacterial Activities of Hygrophila spinosa T. Anders


Arjun Patra1*, Shivesh Jha2, P Narasimha Murthy3 and Aher Vaibhav D1

1College of Pharmacy, IFTM, Moradabad, U.P., India,

2Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi, Jharkhand, India,

3Royal College of Pharmacy and Health Sciences, Berhampur, Orissa, India.

*Corresponding Author E-mail:



Petroleum Ether, chloroform, alcoholic and aqueous extracts of leaves of Hygrophila spinosa were evaluated for anthelmintic and antibacterial activities. The results revealed that the alcoholic extract produced significant anthelmintic activity, both alcoholic and chloroform extracts showed significant antibacterial activity as compared to other extracts. Piperazine citrate and ofloxacin were used as standard drug for anthelmintic and antibacterial activities respectively.


KEY WORDS: Hygrophila spinosa T. Anders, anthelmintic activity, antibacterial activity.




Hygrophila spinosa T. Anders (Acanthaceae)1, commonly known as Talmakhana is widely distributed throughout India, Srilanka, Burma, Malaysia and Nepal. It is a spiny, stout, annual herb and common in water logged places.2 The plant is having low molluscicidal activity against Bulinus truncates and antitumor activity.3, 4 The leaf, root and seed of this plant are being traditionally used for the treatment of rheumatism, inflammation, jaundice, hepatic obstruction, urinary infection, oedema, gout, diabetes, bacterial infection etc. 5, 6, 7 The vast ethnomedical uses insisted us to investigate anthelmintic and antibacterial activities of the leaves of H. spinosa.



Plant Material:

The leaves of H. spinosa were collected from Berhampur, Orissa, India during August to December and authenticated through Birla Institute of Technology, Ranchi, India. A voucher specimen has been deposited at the department of pharmaceutical sciences in the pharmacognosy department in the same institute. The leaves were dried under shade and coarsely powdered.


Preparation of Extract:

The powdered plant material was successively extracted with petroleum ether, chloroform and alcohol using soxhlet apparatus. Finally the aqueous extract was prepared by decoction.


The yield of petroleum ether, chloroform, alcoholic and aqueous extracts were 3, 2.4, 14.2 and 6.2 % respectively. The prepared extracts were tested for anthelmintic and antibacterial activity.


Table 1.  Anthelmintic Activity of Leaves of Hygrophila spinosa


Time Taken in minutes for







Piperizine Citrate

21.2 ± 0.5

26.7 ± 0.6

Petroleum Ether Extract






161 ± 5.7

122 ± 3.5




171.1 ± 12.4

Chloroform Extract





117.3 ± 3.5

103.5 ± 5.0

78.9 ± 3.3


142.0 ± 5.6

123.5 ± 3.7

90.9 ± 4.4

Alcoholic Extract





93.1 ± 4.2

65.8 ± 2.9

51.2 ± 1.6


117.4 ± 2.5

80.4 ± 2.9

62.5 ± 3.1

Aqueous Extract





177.0 ± 6.3

124.3 ± 4.2

68.7 ± 2.6



138.5 ± 2.3

80.5 ± 2.1


Phytochemical Screening:

All the extracts were subjected to preliminary phytochemical screening 8, 9 and it was observed that the petroleum ether extract of leaf contains flavonoids, proteins and fats and oils; chloroform extract of leaf contains

Test Organism

Different Extracts of the Leaf (100mg/disc)

Ofloxacin (5µg/disc)

Petroleum  ether




E. coli NCIM 2341

S. aureus NCIM 2654

B. subtilis NCIM 2195

P. aeruginosa NCIM  2914




























Table 2.  *Observations on Determination of Zone of Inhibition (mm)

* Average of three readings


alkaloids, flavonoids, steroids, proteins and fats and oils; alcoholic extract of leaf contains alkaloids, flavonoids, steroids, tannins and proteins; and aqueous extract of leaf contains tannins, proteins, organic acids and mucilage.


Study of Anthelmintic Activity:

The anthelmintic activity of the extracts of leaf of H. spinosa was determined by using the method of Rao et al.10 The activity was evaluated on adult Indian earthworm (Pheretima posthuma) due to its anatomical and physiological similarity with the intestinal roundworm parasites of human being.11, 12 Fifty milliliters of the extract solution containing three different concentrations, each of petroleum ether, chloroform, alcoholic and aqueous extract (25, 50, 100mg/ml in 1% Tween 80 in normal saline) were prepared and six worms were placed in it. Piperazine citrate (15 mg/ml) was used as reference standard while 1% Tween 80 in normal saline as control. The time taken to paralyse and kill individual worms was observed. Paralysis was noted when the worms became immobile even in the normal saline solution. Death was concluded when the worms lost their motility followed by fading away of their body color.


Study of Antibacterial activity:

The antibacterial activity was evaluated by disc-diffusion method. 13, 14, 15 The bacterial stains used were Escherchia coli (NCIM NO. 2341), Staphylococcus aureus (NCIM NO. 2654), Bacilus subtilis (NCIM NO. 2195) and Pseudomonas aeruginosa (NCIM NO. 2914).  Nutrient agar media was taken in a pre-sterilized Petri-dish and the microorganisms were grown. The extracts were dissolved separately in dimethyl sulphoxide (DMSO) and used in the concentration of 100mg/disc in triplicate, placed in Petri dishes and incubated at 370C for 24 hrs. The diameters of zone of inhibition (mm) were recorded and compared with standard drug ofloxacin.



Preliminary phytochemical investigation showed the presence of organic acids, alkaloids, tannins, mucilage, steroids, flavonoids, proteins and fats and oils in the leaves of H. spinosa..


Table-1 shows that the alcoholic extract at a dose of 100mg/ml has significant anthelmintic activity, whereas chloroform and aqueous extract showed moderate activity


and petroleum ether extract is having least anthelmintic activity.


The results of antibacterial activity (Table-2) indicated that the chloroform and alcoholic extract exhibited significant activity, whereas the aqueous extract has moderate activity and petroleum ether extract is having least activity against the microorganisms.



From the study it can be concluded that the Petroleum Ether, chloroform, alcoholic and aqueous extracts of leaves of Hygrophila spinosa were having both anthelmintic and antibacterial activities. However the active constituents responsible for these activities can be identified.



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Received on 24.07.2008       Modified on 22.08.2008

Accepted on 10.11.2008      © RJPT All right reserved

Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008;Page 531-532