EVALUATION OF ANTIBACTERIAL AND HAEMOLYTIC ACTIVITY OF PHYTOCHEMICALS FROM FRESHWATER MICROALGA, EUGLENA VIRIDIS (EHREN)

  • Jyotirmayee Pradhan Department of Zoology, KKS Womens College, Balasore, Odisha
  • Basanta Kumar Das ICAR-Central Inland Fisheries Research Institute, Barrackpore, Kolkata
  • Swagatika Sahu KVK, Bhanjanagar, Odisha

Abstract

Objective: The main aim of this research work was to evaluate the antibacterial and haemolytic activities of different extracts of Euglena viridis (E. viridis), a freshwater microalga.

Methods: The solvent extraction has been followed by a preliminary screening of phytochemicals. The ethanolic extract, Eu(EtOH) was chromatographed on a silica gel column. The column was eluted with hexane and then with ethyl acetate/hexane mixtures of increasing polarity, 16 fractions (Ef1-Ef16) were collected and grouped according to their TLC (Thin layer chromatography). Antibacterial activities of different fractions of E. viridis against Pseudomonas putida (P. putida) ATCC49828, P. aeruginosa MTCC 35672, Aeromonas hydrophila (A. hydrophila) MTCC 646, ATCC 49140, eleven strains of Staphylococcus aureus (S. aureus) and thirteen strains of Flavobacterium columnare (F. columnare) was done using disc diffusion methods. Haemolytic activity was carried out by using blood agar plate method. The MIC (Minimum inhibitory concentration) values of active fractions were determined by the broth dilution method.

Results: The results showed that the Eu(EtOH) poses significantly (p≤0.5) higher zone of inhibition (14.0±0.28, 13.5±0.28 mm) against FLV8 and FLV9 respectively. Three strains of Flavobacterium (FLV5, FLV6 and FLV10) were highly sensitive (zone size, 17 mm, 17.5 mm) towards 30% EA: Hex chromatographic eluents (Ef11) with lowest MIC values, e. i 60 µg and 30 µg respectively. Two chromatographic fractions, Ef11 and Ef13 were highly effective (zone size, 14.5 mm and 13.5 mm) against S. aureus (SA5) with lowest MIC value (60µg). Haemolytic activities of all the algal extracts were noticed that both Eu(EtOH) and methanolic extract, Eu(MeOH) of Euglena gives negative results.

Conclusion: These findings suggest that the extract obtained from E. viridis have active substances contributing to the increasing antibacterial potential.

Keywords: Alkaloid, Antibacterial activity, Minimum inhibitory concentration, Phytochemical screening, Staphylococcus aureus

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References

1. Brooks GF, Carroll KC, Butel JS. The growth and death of microorganism. In Jawetz, Melinick, Adelberg’s Medical Microbiology. 24th ed Mcgraw-Hill Toronto; 2007. p. 57.
2. Latha S, Kannabiran K. Antimicrobial activity and phytochemicals of Solanum trinobatum Linn. Afr J Biotechnol 2006;5:2402-4.
3. Bhattacharjee M. Pharmaceutically valuable bioactive compounds of algae. Asian J Pharma Clin Res 2016;9:43-7.
4. Abd El-Baky HH, El Baz FK, El-Baroty GS. Evaluation of marine alga Ulva lactuca L. as a source of the natural preservative ingredient. Am Eurasian J Agric Environ Sci 2008;3:434-44.
5. Ehrenberg CG, Neue beobachtungen über blutartige erscheinungen in aegypten, arabien und sibirien, nebst einer uebersicht und kritik der früher bekannten. Annalen Der Physik Und Chemie Ser 1830;28:477-514.
6. Gojdics M. The Genus Euglena. University of Wisconsin Press, Madison. [Includes keys, descriptions, and fig. of all species known at the time]; 1953.
7. Biswas K. Common fresh and brackish water algal flora of India and Burma. Part I. Records of the Botanical Survey of India; 1949. p. 15.
8. Das BK, Pradhan J, Pattnaik K, Samantray BR, Samal SK. Production of antibacterials from the freshwater alga Euglena viridis (Ehern). World J Microbiol Biotechnol 2005;21:45-50.
9. Foltinova P, Lahitova N, Ebringer L. Antimutagenicity in Euglena gracilis. Mutation Res 1994;323:167-71.
10. Nakashima H, Ohshiro Y, Miyano N, Yamamoto N, Ichikawa S, Kondo H, et al. Synergistic inhibition of human immunodeficiency virus type 1 (HIV-1) replication in vitro by sulphated paramylon and 3’-azido-2’,3’-dideoxythymidine (AZT). Lett Appl Microbiol 2008;18:24-6.
11. Das BK, PradhanJ, Sahu S. The effect of Euglena viridis on the immune system of rohu, Labeo rohita. Fish Shellfish Immunol 2009;26:871-6.
12. Das BK, Pradhan J, Mahammad S, Nayak KK. In vitro cytotoxic and antibacterial activity of ethanolic extract of Euglena viridis. Int J Pharma Bio Sci 2012;3:321-31.
13. Barsanti L, Vismara R, Passarelli V, Gualtieri P. Paramylon (b-1,3-glucan) content in wild-type and WZSL mutant of Euglena gracilis effects of growth conditions. J Appl Phycol 2001;13:59-65.
14. Borowitzka MA, Borowitzka LJ. Vitamins and fine chemicals from microalgae. In: Microalgal Biotechnology press syndicate of the University of Cambridge, New York, USA; 1989.
15. Das BK, Pradhan J. Antibacterial properties of selected freshwater microalgae against pathogenic bacteria. Indian J Fish 2010;57:61-6.
16. Pradhan J, Sahu S, Nilima PM. Antibacterial properties of freshwater Microcystis aeruginosa (Kütz) to bacterial pathogen–a comparative study of bacterial bioassays. Indian J Anim Sci 2011;81:1266-7.
17. Pradhan J, Das BK, Sahu S, Mishra BK, Das BK. Traditional antibacterial activity of freshwater microalga Spirulina platensis to aquatic pathogens. Aquacult Res 2012;43:1287-95.
18. Sofowara A. Medicinal plants and traditional medicine in Africa. Spectrum Books Ltd, Ibadan, Nigeria; 1993. p. 289.
19. Trease GE, Evans WC. Pharmacognosy. 15th Edn. Saunders; 2002. p. 214-393.
20. Motl O, Novotny L. Adsorption column chromatography. In: Laboratory Hand Book of Chromatography and Allied Methods. (Ed. by Mikis, O.) Ellis Horwoods Ltd. Chichester; 1979. p. 150-217.
21. Stahl E. Apparatus and general technique in TLC, In: Thin Layer Chromatography. Ed. Stahl E. Springer Verlag, Berlin; 1969. p. 52-85.
22. Chabbert YA. L’ antibiogramme. Sensibilite et resistancedes bacteries aux antibiotiques. De la Tourelle; 1963. p. 257.
23. Izzo AA, DiCarlo G, Biscardi D, De Fusco R. Biological screening of Italian medicinal plants for antibacterial activity. Phytother Res 1995;9:281-6.
24. Alderman D, Smith P. Development of draft protocols of standard reference methods for antimicrobial agent susceptibility testing of bacteria associated with fish disease. Aquaculture 2001;196:211-43.
25. Kumar G, Karthik L, Bhaskara Rao KV. In vitro anti-candida activity of Calotropis gigantea against clinical isolates of Candida. J Pharm Res 2010;3:539-42.
26. Duncan DB. Multiple range and multiple ‘F’ tests. Biometrics 1955;11:1-42.
27. Ayandele AA, Adebiyi AO. The phytochemical analysis and antimicrobial screening of extracts of Olax subscorpioidea. African J Biotechnol 2007;6:868-70.
28. Das BK, Das DP, Pradhan J, Priyadarshinee B, Sahu I, Roy P, Mishra BK. Evaluation of antimicrobial activity and phytochemical screening of ethanolic extract of greater duckweed, Spirodela polyrrhiza. Int J Pharma Bio Sci 2012;3:822-33.
29. Shihabudeen MS, Priscilla HD, Kavitha T. Antimicrobial activity and phytochemical analysis of selected Indian folk medicinal plants. Int J Pharm Sci Res 2010;1:430-4.
30. Dayong S, Xiao F, Jie S, Lijun H, Jiangong S. Steroids from green alga Chaetomorpha basiretorsa setchell. Chinese J Oceanol Limnol 2008;26:415-8.
31. Bhaigyabati T, Kirithika T, Shiny K, Usha K. Phytochemical screening and antioxidant activity of various extracts Ofsargassum muticum. Int J Pharm Res Dev 2011;3:25-30.
32. Lustigman B, Brown C. Antibiotic production by marine algae isolated from the New York/New Jersey Coast. Bull Environ Cont Toxicol 1991;46:329-35.
33. Yuvaraj N, Kanmani P, Satishkumar R, Paari KA, Pattukumar V, Arul V. Extraction, purification and partial characterization of Cladophora glomerata against multidrug-resistant human pathogen Acinetobacter baumannii and fish pathogens. World J Fish Mar Sci 2011;3:51-7.
34. Zeb A, Ali Z, Ahmad T, Abdulmanon A. Physiochemical characteristics of wheat varieties growing in the same and different ecological regions of Pakistan. Pakistan J Biol Sci 2006;9:1823-8.
35. Dhanalakshmi M, Angayarkanni J. Phytochemistry and antibacterial activity of chlorosarcinopsis species. Int J Sci Technol Res 2013;2:315-21.
36. Woldemichael GM, Wink M. Identification and biological activities of triterpenoid saponins from Chenopodium quinoa. J Agric Food Chem 2001;49:2327-32.
37. de Freitas Araujo MG, Hilario F, Vilegas W. Correlation among antioxidant, antimicrobial, hemolytic, and antiproliferative properties of Leiothrix spiralis leaves extract. Int J Mol Sci 2012;13:9260-77.
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Pradhan, J., B. K. Das, and S. Sahu. “EVALUATION OF ANTIBACTERIAL AND HAEMOLYTIC ACTIVITY OF PHYTOCHEMICALS FROM FRESHWATER MICROALGA, EUGLENA VIRIDIS (EHREN)”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 9, Sept. 2018, pp. 98-104, doi:10.22159/ijpps.2018v10i9.28324.
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