Printer Friendly

Larvicidal activity of marine algae, Sargassum swartzii and Chondria dasyphylla, against malaria vector Anopheles stephensi.


Malaria is still a major endemic disease in foci located in south and southeast of Iran. The annual malaria cases have been reported from 66,075 to 6,211 during 1995-2009, indicating the sharp decline of disease. It is unstable with two seasonal peaks mainly in spring and autumn. These areas include the provinces of Sistan and Baluchistan, Hormozgan and Kerman (1). In this part of the country, six anopheline mosquitoes including An. culicifacies, An. stephensi, An. dthali, An. fluviatilis, An. superpictus, and An. pulcherrimus (Diptera: Culicidae) are known to be the malaria vectors and An. sacharovi and An. maculipennis are considered as malaria vectors in northern part of the country (2-7).

Chemical control methods have been applied against either the immature or the adult of malaria vectors. Applying chemical parricides is the most important part of such program. Mosquito control, using chemical larvicides has been performed during the fight against malaria in Iran and still considered as an important part of vector control. Chemical larvicides are now considered as toxic material to fish and other non-target organisms as well as the environment. They are also responsible for increase of insecticide resistance in arthropods. The extract of whole leaf and essential oil of certain plants have been investigated, and showed toxic effect against some public health pests (8-10). Several species of marine algae from coastlines of Iranian islands and Hormozgan province have been reported (11). Marine algae produce different secondary metabolites with a wide range of biological activities (12). Many studies have been achieved on the screening of biological effects of marine organisms and many active compounds were isolated and characterized (13). Red algae from genus Chondria are known as a producer of cyclic polysulfides, terpenoids, amino acids and amines. Domoic acid derivatives with larvicidal and lowering blood pressure activity have been identified in Chondria armata (14). Secondary metabolites with cytotoxic and antitumor activity have been extracted and identified in Sargassum species (15-16). This study was aimed to determine the larvicidal activity of different extracts of S. swartzii and C. dasyphylla, collected from coastlines of the Persian Gulf, southern Iran, against main malaria vector An. stephensi.


Plant material

Brown algae, Sargassum swartzii C. Agardh (Sargassaceae), Chondria dasyphylla (Woodward) C. Agardh (Rodomelacea), were collected from Asaluye-Niband marine protected area of the Persian Gulf in February 2008. The algae were identified by Dr J. Sohrabipour at the Agriculture and Natural Resource Research Center of Hormozgan (herbarium numbers are 20,424, 20,426 respectively) and the voucher specimens were deposited at this center.

Extraction of marine algae

The algae were air-dried in the shade at room temperature and were smashed to make a powder with a mortar and pestle. Each sample of 200 g was extracted with MeOH-[H.sub.2]O (70:30) (5x200 ml) at room temperature. The combined extracts were evaporated under vacuum. The residues were subjected to Silica gel (230) mesh and diluted successively with n-Hexane, CH[Cl.sub.3], EtOAc and Methanol. Removal of the solvents resulted in the production of n-Hexane, CH[Cl.sub.3], EtOAc and MeOH-[H.sub.2]O fractions.

Biological study

Different extracts of S. swartzii and C. dasyphylla were evaluated against late III and early IV instar larvae of An. stephensi. The mosquitoes were collected from malarious areas of Iran, and then were maintained at the insectary of School of Public Health & National Institute of Health Research, Tehran. The reared susceptible larvae to different insecticides were exposed to different concentrations of the S. swartzii and C. dasyphylla extracts which were prepared in methanol. The minimum concentration was 2.5 mg/l and the maximum was 40 mg/l. These concentrations gain the appropriate mortality to plot the regression line. Mortality was determined after 24 h exposure period. All the tests were conducted at 30 [+ or -] 1[degrees]C and 60 [+ or -] 5% relative humidity, and 10 : 14, dark : light periods respectively in the laboratory conditions (17,18). For each concentration, at least 4 replicates of (25) individuals were used (19).

Statistical analysis

The mortality data were subjected to probit analysis using Finney studies (20). From the regression line between logarithmic dose and probit mortality all the parameters including [LC.sub.50] and 95% confidence interval, [LC.sub.90] and 95% confidence interval were determined (21). The regression line was plotted using Microsoft Excel.


Mortality data of An. stephensi exposed to different extracts of two algae, S. swartzii and C. dasyphylla are shown in Table 1. The EtOAc fraction of both S. swartzii and C. dasyphylla were found to be more effective than the other fractions and total extract. Other fractions didn't show significant larvicidal effect against An. stephensi. For EtOAc fractions the chi-square values were significant at p <0.05 level (22). [LC.sub.50] and [LC.sub.90] values for S. swartzii were 11.7584 and 53.472 ppm respectively, and values for C. dasyphylla were 10.625 and 56.394 ppm, respectively (Table 2). The probit regression line is plotted in Fig. 1. From this probit regression line different parameters about efficacy of product against malaria vector can be calculated.


Secondary metabolites with broad range of activities have been found in marine algae. To evaluate the larvicidal effect of the algae from the Persian Gulf against An. stephensi, the samples were extracted with methanol (70%) and fractions were obtained by using various polar and non-polar solvents.


In a previous study on antiplasmodial and antimicrobial activities of South African marine algal extracts, the dichloromethane fraction of Sargassum heterophyllum showed the most antiplasmodial effect with [IC.sub.50] value of 2.8 [micro]g/ml against chloroquine sensitive strain of Plasmodium falciparum (D10) (23).

Exposure of An. stephensi larvae to sub-lethal doses of neem extracts in the laboratory prolonged larval development, reduced pupal weight, high oviposition deterrence and high mortality (24). Some researchers have shown ethanol extract of aerial parts of Tagetes minuta had larvicidal effects with [LC.sub.50] value about 2.5 mg/l (25). Also for Conyza albida, [LC.sub.50] value of 2 mg/l and for Artmisisa afra, [LC.sub.50] of 5 mg/l has been determined (26). In another report for Maytenus senegalensis, [LC.sub.50] value was about 3.9 mg/l and for Harrisonia abyssinica [LC.sub.50] 4.7 mg/l have been reported (27).


In conclusion, larvicidal effects of EtoAc fractions of S. swartzii and C. dasyphylla could be related to semi-polar compounds existing in both algae. The extracts from these plants may be useful for improvement of new natural insecticides, however, further investigations are needed to identify and purify the effective components and their mechanisms of actions of these algae.


This study is a part of Pharm. D. thesis funded and supported by the Tehran University of Medical Sciences



(1.) Manouchehri AV, Zaim M, Emadi AM. A review of malaria in Iran, 1957-1990. J Amer Mosquito Control Assoc 1992; 8(4): 381-5.

(2.) Abai MR, Mehravaran A, Vatandoost H, Oshaghi MA, Javadian E, Mashayekhi M, et al. Comparative performance of imagicides on Anopheles stephensi, main malaria vector in a malarious area, southern Iran. J Vector Borne Dis 2008; 45(4): 307-12.

(3.) Oshaghi MA, Sedaghat MM, Vatandoost H. Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran. East Mediterr Health J 2003; 9(4): 59-66.

(4.) Sedaghat MM, Linton YM, Nicolescu G, Smith L, Koliopoulos G, Zounos AK, et al. Morphological and molecular characterization of Anopheles (Anopheles) sacharovi Favre, a primary vector of malaria in the Middle East. Systematic Entomol 2003; 28: 241-56.

(5.) Sedaghat MM, Harbach RE. An annotated checklist of the Anopheles mosquitoes (Diptera: Culicidae) in Iran. J Vector Ecol 2005; 30: 272-6.

(6.) Zahirnia AH, Vatandoost H, Nateghpour M, Javadian E. Insecticide resistance/susceptibility monitoring in Anopheles pulcherrimus (Diptera: Culicidae) in Ghasreghand district, Sistan and Baluchistan province. Hakim 1998; 1: 97-106.

(7.) Zahirnia AH, Taherkhani H, Vatandoost H. Observation of malaria sporozoite in Anopheles culicifacies (Diptera: Culicidae) in Ghasreghand district, Sistan & Baluchistan province. Hakim 2001; 4: 149-53.

(8.) Hadjiakhoondi A, Sadeghipour-Roodsari HR, Vatandoost H, Khanavi M, Abaee MR, Vosoughi M, et al. Fatty acid composition and toxicity of Melia azedarach L. fruits against malaria vector Anopheles stephensi. Iranian J Pharm Sci 2006; 2(2): 97-102.

(9.) Hadjiakhoondi A, Vatandoost H, Jamshidi A, Bagherj Amiri E. Chemical constituents and efficacy of Cymbopogon olivieri (Boiss) bar essential oil against malaria vector, Anopheles stephensi. Daru 2003; 11(3): 125-8.

(10.) Vatandoost H, Moinvaziri VM. Larvicidal activity of neem tree extract (Neemarin) against mosquito larvae in the Islamic Republic of Iran. Eastern Med Health J 2004; 10: 573-8.

(11.) Sohrabipour J, Rabii R. A list of marine algae of sea shores of the Persian Gulf and Oman Sea in the Hormozgan province. Iran J Bot 1999; 8(1): 131-62.

(12.) Mayer AMS, Rodriguez AD, Berlinck RGS, Hamann MT. Marine pharmacology in 2003-04. Comp Biochem Phys 2007;145(c), 553-81.

(13.) Blunden G. Biologically active compounds from marine organisms. Phytother Res 2001; 15: 89-94.

(14.) Mangala B, Solimabi W. Constituents of Chondria armata. Phytochem 2000; 54(8): 979-81.

(15.) Numata A, Kanbara S, Takahashi C, Fujiki R, Yoneda M, Fujita E, et al. Cytotoxic activity of marine algae and a cytotoxic principle of the brown alga Sargassum tortile. Chem Pharm Bull 1991; 39(8): 2129-31.

(16.) Tang HF, Yi YH, Yao XS, Xu QZ, Zhang SY, Lin HW. Bioactive steroids from the brown alga Sargassum carpophyllum. J Asian Nat Prod Res 2002; 4: 95-105.

(17.) Senthil Nathan S, Kalaivani K, Murugan K, Chung PG. Effects of neem limonoids on malarial vector Anopheles stephensi Liston (Diptera: Culicidae). Acta Trop 2005; 96: 47-55.

(18.) Senthil Nathan S, Kalaivani K, Sehoon K. Effects of Dysoxylum malabaricum Bedd. (Meliaceae) extract on the malarial vector Anopheles stephensi Liston (Diptera: Culicidae). Biores Technol 2006b; 97: 2077-83.

(19.) Instructions for determining susceptibility or resistance of mosquito larvae to insecticides. WHO/VBC-81, 1981; p. 807.

(20.) Finney DJ. Probit analysis. III edn. Cambridge: Cambridge University Press 1971; p. 42-6.

(21.) Cary NC, Saxena SC, Sumithra L. Laboratory evaluation of leaf extract of new plant to suppress the population of malaria vector Anopheles stephensi Liston (Diptera: Culicidae). Curr Sci 1985; 54: 201-2.

(22.) Wandscheer CB, Duque JE, Da Silva MAN, Fukuyama Y, Wohlke JL, Adelmann J, Fontana JD. Larvicidal action of ethanolic extracts from fruit endocarps of Melia azedarach and Azadirachta indica against the dengue mosquito Aedes aegypti. Toxicon 2004; 44: 829-35.

(23.) Lategan C, Kellerman T, Afolayan AF, Mann MG, Antunes EM, Smith PJ, et al. Antiplasmodial and antimicrobial activities of South African marine algal extracts. Pharm Biol 2009; 47(5): 408-13.

(24.) Su T, Mulla MR. Oviposition bioassay responses of Culex tarsalis and Culex quinquefasciatus to neem products containing azadirachtin. Entomol Exp Appl 1999; 91: 337-45.

(25.) Hadjiakhoondi A, Vatandoost H, Khanavi M, Abaee M, Karami M. Biochemical investigation of different extracts and larvicidal activity of Tagetes minuta L. on Anopheles stephensi larvae. Iran J Pharm Sci 2005; 2: 81-4.

(26.) Clarkson C, Maharaj VJ, Crouch NR, Grace OM, Pillay P, Matsabisa MG, Bhagwandin N, Smith PJ, Folb PI. In vitro antiplasmodial activity of medicinal plants native to or naturalized in South Africa. J Pharmacol 2004; 92(2-3): 177-91.

(27.) El Tahir A, Satti G, Khalid S. Antiplasmodial activity of selected Sudanese medicinal plants with emphasis on Maytenus senegalensis (Lam) Exell. J Ethnopharmacol 1999; 64(3): 227-33.

Mahnaz Khanavi [1], Pouyan Bagheri Toulabi [1], Mohammad Reza Abai [2], Nargess Sadati [1], Farzaneh Hadjiakhoondi [1], Abbas Hadjiakhoondi [1] & Hassan Vatandoost [2]

[1] Department of Pharmacognosy and Medicinal Plant Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran; [2] Department of Medical Entomology & Vector Control, School of Public Health & National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran

Correspondence to: Dr Hassan Vatandoost, Department of Medical Entomology & Vector Control, School of Public Health & National Institute of Health Research, Tehran University of Medical Sciences, P.O. Box 6446-14155, Tehran, Iran.


Received: 13 May 2011

Accepted in revised form: 13 December 2011
Table 1. Comparison of larvicidal effect of different extracts
of Sargassum swartzii and Chondria dasyphylla on An. stephensi

                      Concentration    Total    Total   Mortality
Samples                   (ppm)        tested   dead       (%)

Control               70% (methanol)     50       0         0

Sargassum swartzii
Total extract               40           50       4         8
Chloroform                  40           50       0         0
Ethyl acetate               40           52      50       96.1
MeOH                        40           49       3        6.1

Chondria dasyphylla
Total extract               40           50       3         6
Chloroform                  40           49       0         0
Ethyl acetate               40           51      47       92.1
MeOH                        40           50       2         4

Note: The larvae were exposed to a 40 ppm concentration of
different extracts of Sargassum swartzii and Chondria dasyphylla
which were prepared in methanol. Mortality was determined after
24 h exposure period. For each extract, at least 2 replicates
were used.

Table 2. Probit regression line parameters of extract of
S. swartzii and C. dasyphylla against larvae of An. stephensi

Intercept (a)   Slope (b [+ or -] S.E.) [LC.sub.50] in   [LC.sub.90] in
                                        ppm (95% C.L.)   ppm (95% C.L.)

S. swartzii
-2.0854         1.9484 [+ or -] 0.446       4.9945          41.2960
                                        (11.75-33.78)    (53.47-75.35)
C. dasyphylla
-1.8145         1.7680 [+ or -] 0.385       4.5996          42.2081
                                        (10.62-12.46)    (56.39-83.64)

Intercept (a)      [e.sup.2]      [e.sup.2]    p-value
                (heterogeneity)   table (df)

S. swartzii
-2.0854             23.515         7.81 (3)     0.05

C. dasyphylla
-1.8145             18.334         7.81 (3)     0.05
COPYRIGHT 2011 Indian Council of Medical Research
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2011 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Khanavi, Mahnaz; Toulabi, Pouyan Bagheri; Abai, Mohammad Reza; Sadati, Nargess; Hadjiakhoondi, Farza
Publication:Journal of Vector Borne Diseases
Article Type:Clinical report
Date:Dec 1, 2011
Previous Article:Geographical distribution and evaluation of mosquito larvivorous potential of Aphanius dispar (Ruppell), a native fish of Gujarat, India.
Next Article:Plasmodium vivax malaria presenting with skin rash--a case report.

Terms of use | Privacy policy | Copyright © 2018 Farlex, Inc. | Feedback | For webmasters