Artemia salina lethality and histopathological studies on Bacopa monnieri leaf extract

DOI: 10.18805/ijar.v0iOF.7606    | Article Id: B-462 | Page : 610-614
Citation :- Artemia salina lethality and histopathological studies onBacopa monnieri leaf extract .Indian Journal of Animal Research.2018.(52):610-614

Natthawut Charoenphon, Niwat Kangwanrangsan and Wannee Jiraungkoorskul

Address :

Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.

Submitted Date : 15-02-2016
Accepted Date : 10-11-2016


Bacopa monnieri leaf aqueous and methanolic extractions at 1, 3, 5 and 24 hours were determined for the highest amount of total phenolic compound and used for evaluating the toxicity test against Artemia salina at varied concentrations as 0, 5, 50, 100, 500, 1000, 2,500 and 5,000 ppm, by determining the median and 90% lethal concentration, LC50 and LC90, respectively, within 24 hours. The result revealed that the total phenolic compound measurements in 1, 3, 5 and 24 hours were 70.58+2.74, 70.52+1.63, 67.11+2.51 and 79.01+2.87 mg of gallic acid equivalent per gram of aqueous extractions, and 37.88+1.57, 43.08+2.74, 45.86+2.68 and 55.31+3.60 mg of gallic acid equivalent per gram of methanolic extractions, respectively. Due to the highest amount of total phenolic compound, the 24-hour aqueous extract of B. monnieri leaf expressed the 24-h LC50 and LC90 values in A. salina and they were 3,577.90 and 6,440.22 ppm, respectively. Under histological analysis, pathological lesions like cellular swelling, and elongation of the epithelial cells and edema were observed. More additional, cells protruding into the lumen and absence of microvilli were also found. 


Artemia salina Brine shrimp Bacopa monnieri Histopathology Leave Total phenolic compound.


  1. Agrawal, H., Kaul, N., Paradkar. A.R. and Mahadik. K.R. (2006). Separation of bacoside A3 and bacopaside II, major  triterpenoid saponins in Bacopa monnieri, by HPTLC and SFC. Application of SFC in implementation of uniform design for herbal drug standardization, with thermodynamic study. ACTA Chromatogr. 17: 125-150.
  2. Alam, M., Wahed, T. Sultana, F., Ahmed, J. and Hasan, M. (2012). In vitro antioxidant potential of the methanolic extract of Bacopa monnieri L. Turkish J. Pharm. Sci. 9: 285-292.
  3. Charoenphon, N., Anandsongvit, N., Kosai, P., Sirisidthi, K., Kangwanrangsan, N. and Jiraungkoorskul, W. (2016). Brahmi  (Bacopa monnieri): Up-to-date of memory boosting medicinal plant: A review. Indian J. Agri. Res. 50: 1-7.
  4. Conforti, F., Sosa, S., Marrelli, M., Menichini, F., Statti, G., Uzunov, D., et al. (2009). The protective ability of Mediterranean  dietary plants against the oxidative damage: The role of radical oxygen species in inflammation and the polyphenol, flavonoid and sterol contents. Food Chem. 112: 587-594.
  5. Doke, S. and Dhawale, S. (2015). Alternatives to animal testing: A review. Saudi Pharm. J. 23: 223-229. 
  6. D’Souza, P., Deepak, M., Rani, P., Kadamboor, S., Mathew, A., Chandrashekar, A. and Aqarwal, A. (2002). Brine shrimp lethality assay of Bacopa monnieri. Phytotherapy Res. 16: 197-198.
  7. Finney, D.J. (1971). Probit analysis, Cambridge University Press, Cambridge, UK.
  8. Gajardo, G. and Beardmore, J. (2012). The brine shrimp Artemia: Adapted to critical life conditions. Frontiers Physiol. 3: 185.
  9. Gunasekara, R., Rekecki, A., Cornillie, P., Cornelissen, M., Sorgeloos, P., Simoens, P., et al. (2011). Morphological characteristics of the digestive tract of gnotobiotic Artemia franciscana nauplii. Aquaculture 321: 1-7.
  10. Hamidi, M., Jovanova, B. and Panovska, T. (2014). Toxicîlogical evaluation of the plant products using brine shrimp (Artemia salina L.) model. Macedonian Pharm. Bull. 60: 9-18.
  11. Hootman, S. and Conte, F. (1974). Fine structure and fuction of the alimentary epithelium in Artemia salina nauplii. Cell  Tissue Res. 155: 423-436.
  12. Jiraungkoorskul, W. (2016). Larvicidal and histopathological effects of Andrographis paniculata leaf extract against Culex  quinquefasciatus larva. Walailak J. Sci. Technol. 13: 133-140.
  13. Kikuchi, S. (1971). The fine structure of the alimentary canal of the brine shrimp, Artemia salina: the midgut. Ann. Report Iwate Med. Uni 6: 17-47.
  14. Kjanijou, M., Jiraungkoorskul, K., Kosai, P. and Jiraungkoorskul, W. (2012). Effect of Murraya paniculata leaf extract  against Culex quinquefasciatus larva. Asian J. Biol. Sci. 5: 201-208.
  15. Mcdonald, S., Prenzler, P.D., Antolovich, M. and Robards, K. (2001). Phenolic content and antioxidant activity of olive oil extracts. Food Chem. 73: 73-84. 
  16. Meyer, B., Ferrign, N., Putnam, J., Jacobsen, L., Nichols, D. and McLaughlin, J. (1982). Brine shrimp: A convenient  general bioassay for active plant constituents. J. Med. Plant Res. 45: 31-34.
  17. Michael, A., Thompson, C. and & Abramovitz, M. (1956). Artemia salina as a test-organism for bioassay. Sci. 123: 464.
  18. Nunes, B.S., Carvalho, F., Guilhermino, L. and Stappen, G. (2006). Use of the genus Artemia in ecotoxicity testing. Environ.  Poll. 144: 453-462.
  19. Organisation for Economic Co-operation and Development (2002). OECD Guidelines for the Testing of Chemicals / OECD Series on Testing and Assessment Harmonised Integrated Classification System for Human Health and Environmental Hazards of Chemical Substances and Mixtures. Volume 33 of OECD Guidelines for the Testing of     Chemicals / OECD Series on Testing and Assessment. OECD Publishing, Paris, 249p.
  20. Pavananundt, P., Jiraungkoorskul, K., Kosai, P. and Jiraungkoorskul, W. (2013). Larvicidal properties of Cassia siamea  leaf against Aedes aegypti larvae. Int. J. Modern Agri. 2: 1–8. 
  21. Piluzza, G. and Bullitta, S. (2011). Correlations between phenolic content and antioxidant properties in twenty-four plant species of traditional ethnoveterinary use in the Mediterranean area. Pharm. Biol. 49: 240-247.
  22. Rajabi, S., Ramazani, A., Hamidi, M. and Naji, T. (2015). Artemia salina as a model organism in toxicity assessment of  nanopaticles. Daru J. Pharm. Sci. 23: 20.
  23. Shah, M., Behara, Y. and Jagadeesh, B. (2012). Phytochemical screening and in vitro antioxidant activity of aqueous and  hydroalcoholic extract of Bacopa monnieri Linn. Int. J. Pharm. Sci. Res. 3: 3418-3424.
  24. Street, R. and Prinsloo, G. (2013). Commercially important medicinal plants of South Africa: A review. J. Chem. Article ID   205048, 16 pages. doi:10.1155/2013/205048
  25. Subashri, B. and Pillai, Y.J.K. (2014). A comparative study of antioxidant activity of Baccopa monnieri (L.) Pennell using various solvent extracts and its GC-MS analysis. Int. J. Pharm. Pharm. Sci. 6: 494-498.
  26. Surveswaran, S., Cai, Y., Corke, H. and Sun, M. (2007). Systematic evaluation of natural phenolic antioxidants from 133  Indian medicinal plants. Food Chem. 102: 938-953.
  27. Tripathi, N., Chouhan, D.S. Saini, N. and Tiwari, S. (2012). Assessment of genetic variations among highly endangered  medicinal plant Bacopa monnieri (L.) from central India using RAPD and ISSR analysis. 3 Biotech 2: 327-336.
  28. Tupe, R., Kemse, N. and Khaire, A. (2013). Evaluation of antioxidant potentials and total phenolic contents of selected Indian herbs powder extracts. Int. Food Res. J. 20: 1053-1063.
  29. Veeru, P., Kishor, M. and Meenakshi, M. (2009). Screening of medicinal plant extracts for antioxidant activity. J. Med. Plants Res. 3: 608-612.
  30. Verma, M. (2014). Ethnomedicinal and antimicrobial screening of Bacopa monnieri (L.) pennell. J. Phytol. 6: 1-6.
  31. Yanishlieva, N., Marinova, E. and Pokorny, J. (2006). Natural antioxidants from herbs and spices. Eur. J. Lipid Sci. Technol.  108: 776-793.

Global Footprints