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Cytotoxicity of Certain Seed Extracts Against Artemia Fransiscana

Raufun Patoary
Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

Omar Ali Mondal
Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

Wahedul Islam
Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

Ataur Rahman Khan*
Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

The chloroform and methanolic extracts of seed coat and seed kernel of Mucuna pruriens, Terminalia. bellirica, Syzygium cumini and Myristica fragrans were tested against the brine shrimp, Artemia fransiscana nauplii for mortality at 24h post exposure. All the test extracts were found to be effective. The toxicity of the chloroform extracts of seed coat could be arranged in the order: T . bellirica > M. pruriens > S. cumini > M. fragrans. In case of seed kernel extracts (chloroform) the results could be arranged in the following order: S. cumini > M. fragrans > T . bellirica > M. pruriens. For methanolic extracts of seed coat and seed kernel the results could be arranged in the following order: M. fragrans > T . bellirica > S. cumini > M. pruriens, and T . bellirica > S. cumini > M. fragrans > M. pruriens. The brine shrimp bioassay is a simple, reliable and convenient method for the assessment of bioactivity of medicinal plants and lends support for their use in traditional medicine.

Keywords: Seed coat and kernel extracts, Cytotoxicity, Artemia fransiscana .

Location: Taxonomial Section, Department of Botany, University of Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Pest Management

Commodity: Extract (plant, seed)

Objectives:

To evaluate the efficacy of the seed coat and seed extracts as possible sources of potential secondary metabolites to be used as environment friendly pest control agents.

Methodology:

Fresh seeds of M. pruriens (alkushi), T. bellirica (bahara), S. cumini (kalojam) and M. fragrans (jayfal) were collected from the Botanical Garden, Rajshahi University and the identification of voucher specimens were confirmed at the Taxonomical Section, Department of Botany, University of Rajshahi, Bangladesh. Preparation of extract: The seeds were chopped into small pieces, dried under internal shade and powdered using a hand grinder separately. The seed and seed coat powder were extracted with chloroform and methanol (BDH, Pooleg, England) using Soxhlet apparatus according to Feuerhake and Schmutterer. The extracts obtained were stored in a refrigerator at – 20 ⁰C with proper labeling. Preparation of simulated seawater: The eggs of shrimps were collected from the Department of Pharmacy, University of Rajshahi, Bangladesh. Since the lethality test involves the culture of brine shrimp nauplii, i.e., the nauplii should be grown in water with salinity similar to that of seawater. Accordingly, a 3.8% sodium chloride solution was prepared by dissolving 38 gm sodium chloride in 1000 ml distilled water. The P H of the brine water was maintained between 8.0 and 9.0 by using NaHCO₃. Brine water was taken in a small tank and shrimp eggs (1.5 gm/L) were added to one side of the perforated tank with a constant oxygen supply. A constant temperature (35 ±2ºC), sufficient light and air supply were maintained to give the sufficient aeration. After 48 hours, shrimp nauplii were collected and used for the experiment. Preparation and application of doses on A. fransiscana: The chloroform and methanolic extracts of seed coat and kernel of the test plants were applied against the brine shrimp nauplii. For the seed coat and kernel of M. pruriens, T. bellirica, S. cumini and M. fragrans samples were initially dissolved in 200µl of pure dimethylsulfoxide (DMSO) to make them hydrophilic. Clean vials were taken for the 10 samples in five concentrations (two vials for each concentration) and 10 vials were also taken for control test. Five milliliter of seawater containing 10 brine shrimp nauplii was kept in each vial. With the help of a micropipette, specific volumes of samples were transferred from the stock solutions to the vials to get final concentrations of 5.0, 10.0, 20.0, 40.0 and 80.0 µ g/5 ml of brine, because above this concentration cytotoxicity due to DMSO may arise. In the control vials the same volume of DMSO (as in the sample vials) and 5 ml of seawater were taken and 20µl DMSO was added to each vial. Brine shrimp eggs were hatched in simulated seawater to get nauplii. Test samples were prepared by the addition of the requisite amounts of DMSO for obtaining desired concentrations of the test sample. The nauplii were counted by visual inspection and were taken in vials containing 5ml of brine water. Then samples of different concentrations were added to the pre-marked vials with the help of a micropipette. The vials were left for 24 hours and then the nauplii were counted again to find out the cytotoxicity of the test agents and compared to the results with positive control.

Source of Information: Bangladesh J. Zool. 42(2): 133-139, 2014

Article Link (Online Journal): http://www.banglajol.info/index.php/BJZ/article/view/23345/16048

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From this experiment, it is revealed that each of the test samples showed different mortality rates at different concentrations. This significant lethality of several plant extracts to brine shrimp is an indicative of the presence of potent cytotoxic components which warrants further investigation. A perusal of the data shows that all test extracts produced significant mortalities against A. fransiscana nauplii. However, more comprehensive studies are needed in this line.

Knowledge Management: Journal