Journal of Advanced Pharmacy Education & Research
 
 
Review Article
Year : 2017   |  Volume : 7   |  Issue : 1   |  Page : 28-30  

Brine shrimp lethality assay of Sesbania grandiflora ethanolic extract – in vitro study

Priyangaa Sathasivam, T. Lakshmi

Correspondence Address:Department of Pharmacology, Saveetha Dental College, Saveetha University, Chennai, Tamil Nadu, India

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-4040.197331

Abstract  

To evaluate the lethality assay of Sesbania grandiflora using in vitro parameter. Brine shrimp lethality bioassay is a standard method to investigate the cytotoxicity of herbal extract. S. grandiflora commonly known as Agathi in Tamil that appears to have historical usage as an antibacterial and anti-inflammatory agent. A total of 20 nauplii were added into three replicates of each concentration of the plant extract. After 24 h, the surviving brine shrimps were counted, and percentage of mortality was assessed. This study reveals that S. grandiflora leaf possesses significant cytotoxic activity when tested in vitro. From this study, it can be concluded that ethanolic extract of S. grandiflora possessed marked cytotoxic effect.

Keywords: Antibacterial, Brine shrimp, cytotoxicity, Sesbania grandiflora

How to cite this article:

Sathasivam P, Lakshmi T. Brine shrimp lethality assay of Sesbania grandiflora ethanolic extract – in vitro study. J Adv Pharm Edu Res 2017;7(1):28-30.


Introduction   Top

Sesbania grandiflora is commonly known as Agathi keerai in Tamil. It is a popularly used herbal extract. Various literatures prove that there are about 60 global species belonging to genus Sesbania which are commonly found to be grown in Africa, Australia, and Asia. S. grandiflora is commonly found in tropical regions. The vernacular name of S. grandiflora is also known as agusta, bagphal, and agate (Bengali).[1,2] S. grandiflora is a small, erect, fast growing perennial (4-5 m in just 6 months), and sparsely branched tree that reaches 10-15 m in height and diameter up to 12 cm.[3] It is traditionally used for anti-inflammation,[4] antimicrobial activities,[5,6] anticancer,[7] antioxidative activities,[8,9] anti-ulcer activity,[10,11] and immunomodulatory activity,[12] and associated diseases such as hepatic diseases,[13,14] respiratory diseases,[15] and renal diseases.[16] The juice of the bark or root is traditionally used for the treatment of cancer in India.[17] Brine shrimp lethality bioassay is a simple, high throughput cytotoxicity test of bioactive chemicals. It is based on killing the ability of test compounds on a simple zoological organismbrine shrimp (Artemia salina). [18] Brine shrimp lethality bioassay is widely used in the evaluation of toxicity of heavy metals, pesticides, medicines especially natural plant extracts, etc.[19,20] Hence with this regard, we have chosen S. grandiflora leaf against the brine shrimp to evaluate the lethality assay. 

Materials and methods   Top

Plant material

S. grandiflora leaf was obtained as a gift sample from Green Chem Lab, Bengaluru, India.

Brine shrimp lethality assay

The eggs of brine shrimp were procured from Philadelphia, USA. In a small water tank containing seawater, the eggs were incubated for 48 h for hatching. Required light was provided with Philips 40 Watts lamp for 12 h cycle. After 24 h, the larvae were used for the experiments. The nauplii of brine shrimp were challenged in different test tubes containing 10 mL of sea water and 20 larvae. To this, extracts of leaves at different concentrations (10-50 mg/mL) were added. After 24 h, the viability of larvae was observed, and mortality was recorded. Nauplii were considered dead when they were immobile and stayed at the bottom of the test tubes.[21-23] The percent mortality of brine shrimp was calculated with the formula given below.

[% mortality = Number of brine shrimp dead/Number of brine shrimp introduced ×100]

Results Discussion   Top

This study was conducted to evaluate in vitro brine shrimp lethality assay of an ethonolic extract of S. grandiflora and compare the cytotoxicity results with beneficial effects of the plant.

The results obtained from different concentrations of the extract toward brine shrimp lethality assay are given in Table 1. It shows that extract is highly toxic to brine shrimp with an increase in concentration.

It was observed that at a concentration of 10 mg/ml 100% of mortality is seen in brine shrimp in 24 h. At 20 mg/ml concentration, 100% of brine shrimp was killed in 24 h and 30 mg/ml concentration, 100% of the brine shrimp was killed in 24 h. At a concentration of 40 mg/ ml and 50 mg/ml, 100% mortality of brine shrimp was noticed in 24 h, and these concentration prove the larvicidal efficacy of the S. grandiflora ethanolic leaf extract. The results were represented in Table 1 and Figures 1-6.

Conclusion   Top

Its well-known fact that various cell line studies are there to prove the cytotoxicity efficacy of the herbal extract. In those cell lines,

 

4aa1379c-67cc-46d4-991c-7afe0fef7c11.jpg 

Figure 1: Brine shrimp lethality assay of Sesbania grandiflora leaf extract

01f5ceef-31f2-44a2-872c-863f4a06d8b8.jpg

Figure 2: 10 mg/ml concentration

 

25ee9ef3-509e-4f6f-a837-2d66aeefe0ba.jpg

the maintenance is cumbersome and requires specialized media and protocol. Few cell lines are not economical also. Brine shrimp lethality assay has been emerged to prove the cytotoxicity efficacy of the herbal extract which in the recent years are gaining popularity. The procedure is cost effective. In conclusion, S. grandiflora leaf extract revealed that it possess significant cytotoxicity activity against brine shrimp.

Acknowledgement   Top

The authors wish to acknowledge Saveetha Dental College and Hospitals, Chennai, Tamil Nadu, India, for providing all the necessary facilities and encouragement.

ede02909-1960-47ab-a6e3-a2f4ae27c6d0.jpg

Figure 3: 20 mg/ml concentration

                                        

70d2b5f3-a600-4b60-b815-9986768074f4.jpg

Figure 4: 30 mg/ml concentration

25389110-f290-41c1-b480-75a5c8241d0d.jpg

Figure 5: 40 mg/ml concentration

                                        

b20134cc-9115-4eb4-88fb-3b012eab8fa0.jpg

Figure 6: 50 mg/ml concentration

References   Top

1. Orwa C, Mutua A, Kindt R, Jamnadass R, Simons A. Sesbania grandiflora. Agroforestree Database: A Tree Reference and Selection Guide. Agroforestry database 4.0; 2009.

2. Bhoumik D, Dwivedi J. A review on pharmacological activity of Sesbania grandiflora Linn. Columbia J Pharm Sci 2014;1:40-3.

3. Kashyap S, Mishra S. Phytopharmacology of Indian plant Sesbania grandiflora L. J Phytopharmacol 2012;1:63-75.

4. Patil RB, Nanjwade BK, Manvi FV. Effect of Sesbania grandiflora and Sesbania sesban bark on carragennan induce acute inflammation and adjuvant induced arthritis in rats. Int J Pharm Sci 2010;1:75-89.


5. Hasan N, Osman H, Mohamad S, Chong WK, Awang K, Zahariluddin AS. The chemical components of Sesbania grandiflora root and their antituberculosis activity. Pharmaceuticals 2012;5:882-9.

6. Karumari RJ, Sumathi S, Vijayalakshmi K, Balasubramanian SE. Anthelmintic efficacy of Sesbania grandiflora leaves and Solanum torvum fruits against the nematode parasite Ascaridia galli. Am J Ethnomed 2014;1:326-33.

7. Sreelatha S, Padma PR, Umasankari E. Evaluation of anticancer activity of ethanol extract of Sesbania grandiflora (Agati Sesban) against Ehrlich ascites carcinoma in Swiss albino mice. J Ethnopharmacol 2011;134:984-7.

8. Radhika J, Christia R, Jothi G. Effect of the aqueous extract of Sesbania grandiflora Linn in alloxan induced diabetes in albino rats. World J Pharm Res 2014;3:677-85.

9. Doddola S, Pasupulati H, Koganti B, Prasad KV. Evaluation of Sesbania grandiflora for antiurolithiatic and antioxidant properties. J Nat Med 2008;62:300-7.

10. Gowri SS, Vasabtha K. Free radical scavenging and antioxidant activity of leaves from agathi (Sesbania grandiflora) (L.) Pers. Am Eur J Sci Res 2010;5:114-9.

11. Bhalke RD, Giri MA, Anarthe SJ, Pal SC. Antiulcer activity of the ethanol extract of leaves of Sesbania grandiflora (Linn.). Int J Pharm Pharm Sci 2010;2:206-8.

12. Arunabha M, Satish N. Study the immunomodulatory effects of combined extracts of Sesbania grandiflora flowers and Cocculus hirsutus leaves on the circulating antibody response. Am J Phytomed Clin Ther 2015;3:199-208.

13. Roy A, Bhoumik D, Sahu RK, Dwivedi J. Anti-ulcer activity of aqueous extract of Sesbania grandiflora Linn stems in experimental animals. Indian J Res Pharm Biotechnol 2014;2:1254-6.

14. Pari L, Uma A. Protective effect of Sesbania grandiflora against erythromycin estolateinduces hepatotoxicity. Therapie 2003;58:439-43.

15. Tathe PR, Bheemachari, Uplanchiwar V, Modi A, Gahane A, Jain SK, et al. Hepatoprotective activity of fruit extract of Sesbania grandiflora, Pers. Pharmacologyonline 2010;3:423-30.

16. Ramesh T, Begum VH. Effect of Sesbania grandiflora on lung antioxidant defense system in cigarette smoke exposed rats. Int J Biol Chem 2007;1:141-8.

17. Jain R, Jain SK. Screening of in vitro cytotoxic activity of some medicinal plants used traditionally to treat cancer in Chhattisgarh state, India. Asian Pac J Trop Biomed 2011;1:S147-50.

18. Harwing J, Scott PM. Brine shrimp (Artemia salina L.) Larvae as a screening system for fungal toxins. Appl Microbiol 1971;21:1011-6.

19. Price KS, Waggy GT, Conway RA. Brine shrimp bioassay and seawater BOD of petrochemicals. JWater Pollut Control Fed 1974;46:63-77.

20. Sorgeloos P, van der Wielen CR, Persoone G. The use of Artemia nauplii for toxicity tests - A critical analysis. Ecotoxical Environ Saf 1978;2:249-55.

21. Meyer BN, Ferrigni NR, Putnam JE, Jacobsen LB, Nichols DE, McLaughlin JL. Brine shrimp: A convenient general bioassay for active plant constituents. Planta Med 1982;45:31-4.

22. Persoone G. Proceeding of the International Symposium on Brine Shrimp; Artemia salina. Belgium: University Press; 1988. p. 1-3.

23. Sawer IK, Berry MI, Ford JL. Biochemical mutations of brine shrimp. Lett Appl Microbial 2005;40:24-9.

Figures