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Potentiation of Burmese Pink Cassia Cassia Renigera Benth Extracts Against Brine Shrimp Nauplii Artemia Salina (l.) and Lymnaea Acuminata Lamarck

Meherun Nesa
Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Sahadat Hossain
Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Sadia Afrin Rimi
Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Nurul Islam*
Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

Pet. (Petroleum) ether, CHCl3 (Chloroform) and CH3OH (Methanol) extracts of leaves and stem-bark of Cassia renigera Benth. were evaluated against brine shrimp Artemia salina (L.) nauplii and Lymnaea acuminata Lamarck adults through cytotoxicity and molluscicidal activity tests. Except the Pet. ether extract of leaves all the rest were found effective. Against A. salina nauplii the C. renigera leaves gave LC50 values 428.54, 315.20, 240.30, 180.20, 163.22 and 126.54 ppm for the CHCl3 extract and 528.90, 358.21, 270.70, 192.56, 195.63 and 140.20 ppm for the CH3OH extract after 18, 24, 30, 36, 42 and 48 h of exposure respectively. The stem bark gave LC50 values 536.23, 339.43, 271.56, 192.56, 186.58 and 140.20 ppm for Pet. ether extract; 662.82, 368.90, 283.48, 205.04, 189.97 and 156.85 ppm for CHCl3 and 551.60, 368.90, 334.24, 233.07, 215.52 and 140.22 ppm for CH3OH extract, all after 18, 24, 30, 36, 42 and 48 h of exposure respectively. Against L. acuminata the C. renigera leaves gave LC50 values 117.15, 103.30, 91.65, 66.74, 66.74, 52.12 and 49.35 ppm for the CHCl3 extract, and 216.04, 177.94, 149.50, 111.61, 92.34, 59.43 and 52.20 ppm for the CH3OH extract; the stem bark gave LC50 values 105.77, 140.07, 100.62, 95.48, 86.76, 71.47 and 66.40 ppm; 129.37, 111.72, 96.67, 85.28, 77.66, 78.82 and 77.70 ppm, and 134.76, 107.35, 102.26, 94.21, 83.25, 80.70 and 77.00 ppm for Pet. ether, CHCl3 and CH3OH extracts after 12, 18, 24, 30, 36, 42 and 48 h of exposure respectively.

Keywords: Potentiation; Cassia renigera; Extracts; Artemia salina; Lymnaea acuminata.

Location: The Botanical Garden of the University of Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Pest Management

Commodity: Extract (plant, seed)

Objectives:

In this investigation, leaves and stem-barks of the plant C. renigera have been taken into consideration to find out the cytotoxic effects against Artemia salina and molluscicidal activity against Lymnaea acuminata.

Methodology:

2.1. Collection and Preparation of Test Materials: Fresh leaves and stem-barks of the Burmese Pink Cassia C. renigera were collected separately from the Botanical Garden of the University of Rajshahi, Bangladesh. Identification of the target plant was confirmed by the experts on plant taxonomy in the Department of Botany, University of Rajshahi, Bangladesh. Collected plant materials were cleaned properly with the help of a brush and sliced into small pieces. The chips were spread out on wooden-tray to dry. This was done at room temperature under shade avoiding direct sunshine. The materials were rearranged thrice daily to avoid heaping. It needed 10 to 15 days to ready for grinding. Then the plant materials were powdered in an electric grinder machine avoiding additional heat during grinding. The ground materials were placed separately into conical flasks to add solvents in a proportion of 10:1 as solvent and plant dust materials sealed in conical flask (250 ml) to keep on a shaker for 48 h. The Pet. ether, CHCl3 and CH3OH (Merck, Germany) were used (200 g × 600 mL × 2 times) successively while each of which kept 48 h on a shaker. For each of the extracts filtration was done by using filter paper at 24 h interval in the same flask followed by evaporation until the extract was left as a scum. The extracts were removed to glass vials and preserved in a refrigerator at 4°C with proper labeling.

2.2. Collection and Culture of Test Insects The test agent Artemia salina nauplii were emerged from the cysts released in a beaker containing salty water. The nauplii were hatched from the cysts within a period of 24h of age. Another test agent, adult snails of Lymnaea acuminata were collected from the nearby ponds and low-lying submerged areas in the Rajshahi University Campus. The collected snails were reared in pond water in an aquarium under laboratory conditions for the continuous supply of snails of same size and age during experimentation.

2.3. Brine Shrimp Lethality Test First of all, the test tubes were filled with 10 ml of water and marked the lower meniscus with a marker pen and the tubes were made empty. Then, the doses were prepared at five different concentrations in the test tubes by adding a calculated amount of DMSO (dimethylsulfoxide) with the extracts in order to make them hydrophilic prior to add half of the required amount of water in each. The nauplii were counted by visual inspection and were released in each of the test-tubes with the help of a pipette; the additional amount of water was added to fill the test tubes up to the pre-marked level containing 10 ml of water including the test-tube considered as a control batch. Observation of mortality was made after 18, 24, 30, 36, 42 and 48 h of exposure. Doses for all three extracts (Pet. ether, CHCl3 and CH3OH) were 200, 400 and 600 ppm, respectively.

2.4. Molluscicidal Activity Test From the aquarium culture adult L. acuminata snails were collected. In each of the 50 ml beakers ten individuals were kept. Test samples at different concentrations considered as doses were prepared in beakers by adding calculated amount of DMSO (Dimethylsulfoxide) to make them hydrophilic before adding half of the required amount of water in each. After that a certain amount of pond water was added to fill up the pre-marked beakers with the help of a pipette. The snails were counted by visual inspection and were released in the beakers containing 10 ml of water. Observation of mortality was made after 12, 18, 24, 30, 36, 42 and 48 h of exposure with an interval of 6 h. After experimenting pilot doses, the final applicable doses for leaf extracts were fixed as 50, 60 and 70 ppm; for stem-bark extracts the fixed doses were 70, 80 and 90 ppm. All the experiments were carried out in the Crop Protection & Toxicology Laboratory of the Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

2.5. Statistical Analysis The mortality (%) was corrected using Abbott’s formula (Abbott, 1925). The data were then subjected to Probit analysis by using GW Basic software (Busvine, 1971; Finney, 1947).

Source of Information: Rajshahi University Journal of Environmental Science, 8: 121-126, 2019

Article Link (Online Journal): www.ru.ac.bd/ies

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

After analyzing the results of this investigation and considering the outcome of the previous works done on C. renigera and its relative species, it could be concluded that further developments on the secondary metabolites of this could be used in the pest control strategy. Further investigation should be taken into consideration to explore the exact compound for which these plant extracts show pesticidal effects.

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