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Evaluation of antiplasmodial properties of a cyanobacterium, Spirulina

platensis and its mechanism of action

Article  in  Natural Product Research · August 2017

DOI: 10.1080/14786419.2017.1360880

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 Natural Product Research
Formerly Natural Product Letters

ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20

Evaluation of antiplasmodial properties of a

cyanobacterium, Spirulina platensis and its
mechanism of action

Diah Anggraini Wulandari, Elizabeth Sidhartha, Iriani Setyaningsih, Jonathan

Marshall Marbun, Din Syafruddin & Puji Budi Setia Asih

To cite this article: Diah Anggraini Wulandari, Elizabeth Sidhartha, Iriani Setyaningsih, Jonathan
Marshall Marbun, Din Syafruddin & Puji Budi Setia Asih (2017): Evaluation of antiplasmodial
properties of a cyanobacterium, Spirulina platensis and its mechanism of action, Natural Product
Research, DOI: 10.1080/14786419.2017.1360880

To link to this article: http://dx.doi.org/10.1080/14786419.2017.1360880

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Download by: [University of Notre Dame] Date: 08 August 2017, At: 04:56
 Natural Product Research, 2017
https://doi.org/10.1080/14786419.2017.1360880

SHORT COMMUNICATION

Evaluation of antiplasmodial properties of a cyanobacterium,

Spirulina platensis and its mechanism of action
Diah Anggraini Wulandaria*, Elizabeth Sidharthab*, Iriani Setyaningsiha,
Jonathan Marshall Marbunb, Din Syafruddinb,c  and Puji Budi Setia Asihb 
a
Faculty Fisheries and Marine Science and Tehnology, Department of Aquatic Products Technology, Bogor
Agricultural University, Bogor, Indonesia; b Malaria and Vector Resistance Laboratory, Eijkman Institute
of Molecular Biology, Jakarta, Indonesia; cFaculty of Medicine, Department of Parasitology, Hasanuddin
University, Makassar, Indonesia
Downloaded by [University of Notre Dame] at 04:56 08 August 2017

ABSTRACT ARTICLE HISTORY

The rapid emergence of antimalarial drug resistance necessitates a Received 20 February 2017
continual effort on novel drug discovery. A cyanobacterium, Spirulina Accepted 10 July 2017
platensis, is a potential antimalarial agent that has been widely
KEYWORDS
consumed as food supplement in the form of crude extract. It is Antiplasmodial;
known to possess antiviral, antibacterial and antifungi activities. This cyanobacterium; in vitro;
study examined the antimalarial activities of several Spirulina formulas mechanism of action;
against Plasmodium falciparum 3D7, in vitro. The tested Spirulina Spirulina platensis
formulas included commercially available capsule, crude extract and
alkaloid fraction. Results showed that all tested formula possessed
antimalarial activities with the Spirulina capsule exhibited the highest
activities (IC50 = 2.16 μg/mL). Light and electron microscopies revealed
interference of the Spirulina with the parasite hemozoin formation.
In conclusion, all tested Spirulina formulas and fraction exhibited
moderate to high antimalarial activities.

1. Introduction
Provision of anti-malarial drugs along with vector control is the mainstay of malaria control
efforts for decades. Artemisinin-based combination therapy (ACT), the current first line drug
that was introduced by WHO since the year 2000 has greatly contributed to the significant

CONTACT  Puji Budi Setia Asih  puji@eijkman.go.id

*
These authors contributed equally to this work.
 Supplemental data for this article can be accessed at http://doi.org/10.1080/14786419.2017.1360880.
© 2017 Informa UK Limited, trading as Taylor & Francis Group
 2   D. A. WULANDARI ET AL.

reduction of the malaria burden in many countries worldwide. However, the emergence and
rapid spread of the parasite strains that are resistant to artemisinin in Greater Mekong
Subregion has raised concern and that increases the need for novel drugs against malaria.
Exploration of the natural products that have been empirically used as traditional medicine
offers several advantages which include a reduced or absence of toxicity. The cyanobacte-
rium Spirulina platensis has been reported to exhibit antipathogen activities such as antiviral
(Mader et al. 2016), antibacterial (El-Sheekh et al. 2014), antioxidant (Bashandy et al. 2016),
anti-inflammation (Abdel-Daim et al. 2015) and high protein source (Casio et al. 2017).
S. platensis contains active compounds against malaria, namely phycocyanin and alkaloid
and hence has the potential to be developed as a novel antimalarial drug. Phycocyanin
isolated from Nostoc spp., another cyanobacterium, has been reported to exhibit antimalarial
activity at a concentration 3.0 μg/mL, with IC50 of 10.27 ± 2.79 and 10.37 ± 1.43 μg/mL against
chloroquine-sensitive and chloroquine-resistant Plasmodium falciparum, respectively (Pankaj
Downloaded by [University of Notre Dame] at 04:56 08 August 2017

et al. 2010). Similarly, alkaloids extracted from the plant Albertisia villosa exhibit antimalarial
with IC50 of 73 nM against P. falciparum (Lohombo-Ekomba et al. 2004). This study aims to
determine the antimalarial activity of several Spirulina formulas including commercially
available Spirulina capsule, crude extract and alkaloid fraction.

2.  Results and discussion

A phytochemical analysis of the three formulas of Spirulina; commercially available capsule,
crude extract and the alkaloid fraction containing steroid and saponin (Table S1), demon-
strated antimalarial activity in vitro. Testing revealed that the Spirulina capsule exhibited an
antimalarial activity with an IC50 of 2.16 μg/mL. The crude extract and alkaloid fraction
demonstrated lower antimalarial activity (Table S2). The stronger antimalarial activity of
Spirulina capsule may be linked to its higher alkaloid contents as compared to the other
Spirulina-derived formulations. Alkaloids have been reported to exhibit antimalarial activities
particularly against Plasmodium falciparum. Alkaloids extracted from the plant Albertisia
papuana demonstrated potent antimalarial activities (Lohombo-Ekomba et al. 2004). The
alkaloids isoquinoline Dehaasia longipediqulata and carpaine papaya leaves were reported
to inhibit Plasmodium with IC50 0.031–30.4 μM (Zahari et al. 2014). The steroid derivatives
thiopurine and triazole were reported to inhibit Plasmodium growth with an IC50 of 13.9–
22.8 μM (Corrales et al. 2011). It is likely that the antimalarial activity of Spirulina capsules
and crude extract resulted from a combination of synergistic effect of several active com-
pounds including alkaloids, steroids and saponin. Examination of the exposed parasites
under light microscopy revealed that parasites treated with Spirulina capsules, crude extract
and active fraction at their respective IC50 levels for 48 h exhibited abnormal minute con-
densed structures in the ring form, with a darker stained nucleus (Figure S1). Observation
under transmission electron microscope (TEM) revealed several trophozoites that lacked
hemozoin crystals and indistinct membrane structure. These effects were also observed in
the parasites treated with artemisinin as a positive control. In contrast, the untreated parasites
(negative control) showed normal morphology with intact cell membranes. Figure S2 depicts
ultrastructural morphology of parasites treated with the Spirulina capsule at IC50 for 24 h.
Treated parasites displayed indistinct cell and organelle membranes, as well as a lack of
hemozoin crystals. However, untreated parasites exhibited normal morphology distinct
organelles with intact cell membranes, and hemozoin crystals in digestive vacuoles
 NATURAL PRODUCT RESEARCH   3

(Figure S2). Under normal conditions, the Plasmodium parasite internalises haemoglobin
from its host erythrocyte by endocytosis, and proceeds to hydrolyse the hemoglobin in the
digestive vacuole to produce hame and globin. The toxic hame byproduct is then polymer-
ised into non-toxic hemozoin crystals, which were observed under light microscopy and
electron microscopy in the control arm of study. Antimalarial drugs such as chloroquine
disrupt this process by interfering with hemozoin formation and increasing the pH of the
digestive vacuole. It is possible that the mechanism of antiplasmodial action by Spirulina
capsule is similar to that of chloroquine.

3. Conclusion
Spirulina formulas (commercially available capsule, crude extract and alkaloid fraction) exhib-
ited moderate to high antimalarial activities, through interference with the parasite hemozoin
Downloaded by [University of Notre Dame] at 04:56 08 August 2017

formation.

Supplementary material
Experimental details, figures and tables are available online as supplementary materials.

Acknowledgements
The authors wish to thank the Eijkman Institute for Molecular Biology (EIMB) and Bogor Agricultural
University (IPB) for the support and encouragement. We thank to Ismail E. Rozi for the TEM image and
Dr Neil F Lobo from Notre Dame University, USA, for critical reading of the manuscript. This study is
part of Diah A. Wulandari Master Program in IPB.

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
This study was supported by Research Grant from The Ministry of Research, Technology, and Higher
Education Republic of Indonesia, through EIMB and IPB.

ORCID
Din Syafruddin   http://orcid.org/0000-0001-7141-2545
Puji Budi Setia Asih   http://orcid.org/0000-0002-4582-9133

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 4   D. A. WULANDARI ET AL.

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