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World Journal of Microbiology & Biotechnology 2005 21: 45–50

DOI: 10.1007/s11274-004-1555-3
Springer 2005

Production of antibacterials from the freshwater alga Euglena viridis (Ehren)

B.K. Das*, J. Pradhan, P. Pattnaik, B.R. Samantaray and S.K. Samal

Aquatic Animal Health Division, Central Institute of Freshwater Aquaculture (CIFA), P.O. Kausalyagnaga,
Bhubaneswar 751002, Orissa, India
*Author for correspondence: Tel.: +91-674-2465446, Fax: +91-674-2465407, E-mail: basantadas@yahoo.com

Received 28 January 2004; accepted 20 May 2004

Keywords: Aeromonas hydrophila, bio control, Euglena viridis, organic extract, Pseudomonas, Vibrio

Summary

The antibacterial properties of Euglena viridis, collected from a freshwater pond at the Central Institute of
Freshwater Aquaculture (CIFA), Bhubaneshwar, India, were tested against various strains of virulent pathogens
viz. Pseudomonas putida (PP1, PP2), P. aeruginosa (PA1, PA2, PA3, PA4), P. fluorescens (PF1, PF2, PF3, PF4),
Aeromonas hydrophila (AH30, AH31, AH32, AH34), Edwardsiella tarda, Vibrio alginolyticus (VA1), V. anguillarum
(VN1, VN2 & VN3), V. fluvialis (VF1), V. parahemolyticus (VP1) and V. harveyi (VH1) and Escherichia coli (O115,
O1, O156, O164, O111 & O109). Four organic extracts viz. methanolic, ethanolic, acetone and acetone/ethanol of
the E. viridis showed moderate to high antibacterial activity to all the bacterial pathogens. Rotavapor extraction
products showed higher sensitivity in comparison to cold and hot extractions.

Introduction Edwardsiella tarda, Vibrio alginolyticus, v. anguillarum,

Freshwater microalgae comprise a vast group of pho-
tosynthetic, heterotrophic organisms, which have an
extraordinary potential for cultivation as energy crops.
They are able to produce a wide range of commercially
interesting byproducts such as fats, oils, sugars and
functionally bioactive compounds. Discovering new
therapeutic molecules is becoming increasingly impor-
tant as more and more bacteria become resistant to the
commonly used antibiotics. Traditionally used in Asiatic
medicines, algae, since the second half of the 20th
century, have been screened for their biological activi-
ties. Thus, antibacterial effects have been noticed in all
the algal classes and notably in diatoms, the major
component of the phytoplankton (Duff et al. 1966;
Cooper et al. 1983; Reichelt & Borowitska 1984; Viso
et al. 1987). However, most of these antibiotic activities
have only been tested against human pathogens and the
active molecule(s) have rarely been purified. The anti-
bacterial activities of marine algae have been well
documented (Vora & Babu 1994; Padmakumar 1997;
Naviner et al. 1999). However, a few studies have been
initiated in freshwater algae (Moore et al. 1996).
The present preliminary investigation was carried out
to examine the production of antimicrobial/bioactive
compounds from freshwater Euglenoids by using mi-
crobially based assay systems. In this preliminary work
we have investigated the effects of organic extracts of
Euglena viridis (Ehren) against different strains of fish
and shellfish pathogens, e.g. Aeromonas hydrophila,
Pseudomonas putida, P. aeruginosa, P. fluorescens,
V. harveyi, V. fluvialis, and V. parahaemolyticus and
animal isolates of Escherichia coli.
Materials and methods
Euglena viridis blooms were collected from the ponds
(0.2–2 ha) of the Central Institute of Freshwater Aqua-
culture (CIFA), Kausalyaganga, Bhubaneswar, India in
month of September 2002 with the help of a plankton net
made of bolting silk cloth. The samples were simulta-
neously (4%) preserved in formalin for identification and
identified as E. viridis as per the protocols cited in the
Records of the Botanical Survey of India (Biswas 1949).
Collected samples were washed three times in sterile
filtered water in order to remove suspended particles
adhered to it and finally centrifuged at 1000 · g using
macro rotor (Sorvall CE, UK). The pellets were har-
vested and 4.0 kg of E. viridis (w/w) was dried under
room temperature for 2–3 days. The dried weight of the
algae was taken and found to be 2.46 kg. Then it was
powdered. Five grams of dry euglena powder were taken
for each solvent extraction. Eight samples were taken, to
which different solvents were added.
Solvent extraction
Chloroform, methanol, ethanol, acetone and acetic acid
were used for solvent extraction. Three types of extrac-
tion, i.e. cold extraction, hot extraction and Rotavapor
extraction were followed.

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