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Larvicidal activities of some Euro-Asiatic

plants against Culex quinquefasciatus Say
(Diptera: Culicidae)

Article in Parasitology Research · June 2009

DOI: 10.1007/s00436-009-1511-0 · Source: PubMed

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Parasitol Res (2009) 105:887–892
DOI 10.1007/s00436-009-1511-0

SHORT COMMUNICATION

Larvicidal effects of some Euro-Asiatic plants against

Culex quinquefasciatus Say larvae (Diptera: Culicidae)
Roman Pavela

Received: 27 February 2009 / Accepted: 25 May 2009 / Published online: 5 June 2009
# Springer-Verlag 2009

Abstract Methanol extracts of the aerial parts from 62 environmental threat these chemicals pose, the effects on
Euro-Asiatic plant species were tested for larvicidal activity non-target organisms, and the resistance of mosquitoes to
against the mosquito Culex quinquefasciatus Say (Diptera: insecticides have all increased during the last five decades
Culicidae) under laboratory conditions. The fourth larval (VCRC 1989, Severini et al. 1993, Rao et al. 1995). There
instar was tested. The extracts of the plants Seseli pallasii is an urgent need to develop new materials for controlling
and Schisandra chinencis displayed the highest larvicidal mosquitoes in an environmentally safe way, using biode-
activities with LD50 6 and 15 ppm, respectively, followed gradable and target-specific insecticides against them.
by Arthemisia campestris, Verbena officinalis, and Imper- Plants have been evolving for over 400 million years and
atoria osthruthium with LD50 23, 38, and 49 ppm, have developed protection mechanisms, such as repellents
respectively. The appraised value of LD50 for two species and even insecticidal effects, to defend themselves against
was between 51–100 ppm; eight species had lethal doses insect attack (Andor 1995). The fact that plants have
from 101 to 500 ppm, 13 species showed lethal doses from developed, through evolution, an effective defense system
501 to 1,000 ppm, and 34 species did not show lethal doses against most insects makes them the richest natural source
for low mortality (LD50 >1,000 ppm). of biocidal compounds, such as toxicity (Pavela 2006),
antifeedants (Wheeler and Isman 2001; Pavela 2004a),
insect growth regulators (Akhtar and Isman 2004; Pavela
Introduction 2004b, Pavela 2005), oviposition deterrence (Dimock and
Renwick 1991; Zhao et al. 1998), suppression of calling
Insect-transmitted disease remains a major cause of illness behavior (Khan and Saxana 1986), and reduction of
and death worldwide. Mosquitoes alone transmit disease to fecundity and fertility (Pavela et al. 2005). Such a wide
more than 700 million people annually (Aregawi et al. variety of effects provide potential natural alternatives to
2008). Therefore, the control of mosquitoes is an important the use of synthetic chemical insecticides. Certain plant
public health concern around the world. For example, Culex families, particularly Maliaceae, Asteraceae, Rutaceae,
quinquefasciatus Say (Diptera: Culicidae) is a pantropical Labiaceae, Annonaceae, and Canellaceae, are viewed as
pest and urban vector of Wuchereria bancrofti, Plasmodium exceptionally promising sources of plant-based insecticides
(avian malaria), myxomatosis, and other diseases in some (Pavela 2007). Nevertheless, the current worldwide trend of
parts of the world (Holder 1999). increasing the proportion of biological pesticides forces us
Chemical control is an effective strategy used extensive- to seek new, environmentally safe substances to apply in
ly in daily life. Synthetic insecticides are today in the producing safe new botanical insecticides.
forefront of mosquito-controlling agents. However, the For this reason, it is necessary to test those plant extracts
which could be expected to contain substances of environ-
R. Pavela (*) mental potential and, at the same time, provide adequate
Crop Research Institute,
efficacy against mosquitoes (Trongtokit et al. 2008).
Drnovská 507,
Prague 6 161 06, Czech Republic Screening of the insecticide efficacy of wild plants in the
e-mail: [email protected] Euro-Asian region was performed as part of an international
888 Parasitol Res (2009) 105:887–892

Table 1 Plants used in this study, their part used, origin, voucher references and yield of extracts

Species Family Plant part Yield (%) Voucher Origin

assayed references

Ailanthus altissima (Mill.) Swingle Simaroubaceae Leaves 15.3 907 Prague, Czech Republic
Alexitoxicon scandens Somm. et Lever Asclepiadaceae Stem 10.2 855 Krasnodarskiy region, Russia
Angelica archangelica L. Apiaceae Fruit 4.7 809 Prague, Czech Republic
Anthemis tinctoria L. Asteraceae Flower 5.1 945 Prague, Czech Republic
Arctium lappa L. Asteraceae Stem 7.3 917 Ljulin Mountain, Bulgaria
Artemisia abrotanum L. Asteraceae Stem 8.3 919 Prague, Czech Republic
Artemisia campestris L. Asteraceae Stem 6.9 910 Ljulin Mountain, Bulgaria
Astragalus chinensis L. f. Fabaceae Stem 7.7 944 Prague, Czech Republic
Balsamita major Desf. Asteraceae Stem 7.5 930 Prague, Czech Republic
Borago officinalis L. Boraginaceae Stem 3.9 835 Prague, Czech Republic
Bryonia dioica Jacq. Cucurbitaceae Stem 6.8 940 Prague, Czech Republic
Carthamnus lanatus L. Asteraceae Stem 7.9 916 Ljulin Mountain, Bulgaria
Cichorium intybus L. Asteraceae Stem 5.9 911 Ljulin Mountain, Bulgaria
Clematis vitalba L. Ranunculaceae Stem 8.2 912 Ljulin Mountain, Bulgaria
Convolvulus lineatus L. Convolvulaceae Stem 9.3 854 Krasnodarskiy region, Russia
Dictamnus albus L. Rutaceae Rhizome 2.6 806 Prague, Czech Republic
Dorycnium graeeum (L.) Ser. Fabaceae Stem 6.1 850 Krasnodarskiy region, Russia
Dracocephalum moldavicum L. Lamiaceae Stem 5.8 929 Prague, Czech Republic
Echinacea purpurea (L.) Moench Asteraceae Root 8.1 808 Prague, Czech Republic
Echinops sphaerocephalus L. Asteraceae Stem 8.9 915 Ljulin Mountain, Bulgaria
Ferula assa-foetida L. Apiaceae Stem 3.5 926 Prague, Czech Republic
Galium verticillatum Danth. Rubiaceae Stem 6.9 853 Krasnodarskiy region, Russia
Gilia capitata Sims Polemoniaceae Stem 8.1 816 Prague, Czech Republic
Grindelia camporum Greene Asteraceae Stem 6.9 939 Prague, Czech Republic
Hypericum perforatum L. Clusiaceae Flower 7.8 831 Planice, Czech Republic
Imperatoria ostruthium L. Apiaceae Root 11.1 805 Prague, Czech Republic
Inula auriculata Boiss. & Balansa Asteraceae Stem 4.7 852 Krasnodarskiy region, Russia
Jacobaea maritima (L.) Pelser & Meijden Asteraceae Stem 4.8 833 Prague, Czech Republic
Lapasana intermedia Bieb. Asteraceae Stem 4.5 844 Krasnodarskiy region, Russia
Lobelia siphilitica L. Campanulaceae Stem 5.6 937 Prague, Czech Republic
Lythrum salicaria L. Lythraceae Stem 9.9 924 Prague, Czech Republic
Melilotus albus Medik. Fabaceae Stem 8.9 953 Prague, Czech Republic
Onobrychis miniata Stev. Fabaceae Stem 6.5 841 Krasnodarskiy region, Russia
Onopordon acanthium L. Asteraceae Stem 9.3 914 Ljulin Mountain, Bulgaria
Physalis alkekengi L. Solanaceae Stem 6.9 935 Prague, Czech Republic
Polygala comosa Schk. Polygonaceae Stem 17.5 851 Krasnodarskiy region, Russia
Potentila erecta L. Rosaceae Stem 3.6 846 Krasnodarskiy region, Russia
Psoralea bituminosa L. Fabaceae Stem 13.3 856 Krasnodarskiy region, Russia
Pyrethrum corymbosum (L.) Willd. Asteraceae Flower 4.1 840 Krasnodarskiy region, Russia
Reseda odorata L. Polemoniaceae Stem 7.9 819 Prague, Czech Republic
Reynoutria × bohemica Chrtek & Chrtková Polygonaceae Leaves 11.8 901 Prague, Czech Republic
Reynoutria sachalinensis F. Schmidt Polygonaceae Leaves 12.6 904 Prague, Czech Republic
Rubia tinctorum L. Lamiaceae Stem 8.5 942 Prague, Czech Republic
Salvia glutinosa L. Lamiaceae Stem 4.1 941 Prague, Czech Republic
Saponaria officinalis L. Caryophyllaceae Stem 6.3 943 Prague, Czech Republic
Saponaria officinalis L. Caryophyllaceae Rhizome 8.7 859 Planice, Czech Republic
Scrophularia nodosa L. Scrophulariaceae Stem 5.6 936 Prague, Czech Republic
Sedum rosea (L.) Scop. Crassulaceae Flower 11.3 950 Prague, Czech Republic
Parasitol Res (2009) 105:887–892 889

Table 1 (continued)

Species Family Plant part Yield (%) Voucher Origin

assayed references

Seseli pallasii Besser Apiaceae Stem 6.1 927 Prague, Czech Republic
Schisandra chinensis (Turcz.) Baill Schisandraceae Leaves 8.7 928 Prague, Czech Republic
Silphium perfoliatum L. Asteraceae Leaves 8.7 905 Prague, Czech Republic
Stachys byzantina K.Koch Lamiaceae Stem 11.2 938 Prague, Czech Republic
Symphytum officinale L. Boraginaceae Rhizome 9.1 860 Planice, Czech Republic
Teucrium hircanicum L. Lamiaceae Stem 12.6 946 Prague, Czech Republic
Teucrium chamaedrys L. Lamiaceae Stem 8.3 925 Prague, Czech Republic
Thymus fragrantissimus Samen. Lamiaceae Stem 3.6 920 Prague, Czech Republic
Thymus serphyllum L. Lamiaceae Stem 2.3 921 Prague, Czech Republic
Trifolium arvense L. Fabaceae Stem 13.6 858 Krasnodarskiy region, Russia
Tropaeolum majus L. Tropaeolaceae Leaves 7.2 832 Prague, Czech Republic
Verbena hirta Spreng. Verbenaceae Stem 5.2 955 Prague, Czech Republic
Verbena officinalis L. Verbenaceae Stem 5.4 810 Prague, Czech Republic
Vincetoxicum hirundinaria Medik. Asclepiadaceae Stem 8.2 934 Prague, Czech Republic
Xeranthemum cylindraceum Smith. Asteraceae Stem 4.3 848 Krasnodarskiy region, Russia

project currently being carried out. Plant extracts obtained 28±2°C, 70±5% RH, and a photo regime of 16:8 (L:D) h.
from 56 species, sampled at localities in the Czech Republic Early fourth instar larvae were used in the study.
and Russian Federation, have been assessed for their larvicidal
efficacy against C. quinquefasciatus (Pavela 2008). Larvicidal activity
In this study, we are introducing additional results of the
larvicidal efficacy of extracts, obtained from 62 other Mosquito larvicidal trials were carried out according to
species of plants gathered at various locations in the Czech WHO standard procedures (1996), with slight modifica-
Republic and Russia, against C. quinquefasciatus Say tions. The extracts were diluted in dimethyl sulphoxide
(Diptera: Culicidae). (DMSO) in order to prepare a serial dilution of test dosage.
For experimental treatment, 1 ml of serial dilution was
added to 224 ml of distilled water in a 500-ml glass bowl
Materials and methods and shaken lightly to ensure a homogeneous test solution.
The larvae were transferred in distilled water into a bowl of
Plant extracts prepared test solution with a final surface area of 125 cm2
(25 larvae/beaker). Four duplicate trials were carried out for
Fresh plant material from each of the selected species (see every sample concentration, and for each trial a negative
Table 1) was collected in 2008. Voucher specimens of all control was included using distilled water containing the
the plant species studied were deposited in the respective same amount of DMSO as the test sample. A different
herbaria of our institute. The plant material was shade dried series of concentrations (resulting from the previous
and powdered. The dry powder was extracted with an screening) was used for each extract in order to obtain
excess of methylalcohol (500 ml of MeOH for 100 g of mortality ranging between 10% and 90%. At least five
plant powder) for 24 h. The crude extracts were separately concentrations were selected for the calculation of lethal
filtered and evaporated under reduced pressure in a rotary doses. A dose of 1,000 ppm was determined to be the
evaporator. uppermost limit for ascertaining the larvicidal activity of
extracts. The trials were placed in a growth chamber (L16:
Test organisms D9, 26°C). Mortality was determined after 24 h of
exposure, during which no food was offered to the larvae.
A laboratory colony of C. quinquefasciatus was used for The control mortality was corrected by Abbott's formula
the larvicidal activity. The larvae were fed on dog biscuits (Abbott 1925) and an LC50, LC90 regression equation, and
and yeast powder in a 3:1 ratio. Adults were provided with a 95% confidence limit was calculated using probit analysis
a 10% sucrose solution and blood. Mosquitoes were held at (Finney 1971).
890 Parasitol Res (2009) 105:887–892

Table 2 Larvicidal activity of extracts from Euro-Asiatic plants against larvae of Culex quinquefasciatus after 24 h of exposure

Species Mortality % (mean±SE)a LD50b (CI95) LD90b (CI95) Chic

Ailanthus altissima (Mill.) Swingle 3.3±1.5 >1,000 >1,000

Alexitoxicon scandens Somm. et Lever 15.2±3.2 >1,000 >1,000
Angelica archangelica L. 100.0±0.0 60 (52–68) 146 (121–191) 1.952
Anthemis tinctoria L. 100.0±0.0 396 (338–452) 628 (595–6,388) 3.830
Arctium lappa L. 11.6±2.8 >1,000 >1,000
Artemisia abrotanum L. 100.0±0.0 609 (551–653) 836 (780–893) 3.188
Artemisia campestris L. 100.0±0.0 23 (21–24) 36 (32–42) 1.719
Astragalus chinensis L. f. 38.3±6.2 >1,000 >1,000
Balsamita major Desf. 100.0±0.0 732 (703–761) 891 (849–951) 0.875
Borago officinalis L. 95.2±3.8 371 (289–396) 915 (900–941) 1.486
Bryonia dioica Jacq. 100.0±0.0 112 (79–130) 320 (310–335) 2.885
Carthamnus lanatus L. 25.7±5.2 >1,000 >1,000
Cichorium intybus L. 3.3±1.1 >1,000 >1,000
Clematis vitalba L. 3.3±1.3 >1,000 >1,000
Convolvulus lineatus L. 5.1±0.3 >1,000 >1,000
Dictamnus albus L. 60.0±3.3 982 (970–996) >1,000 3.356
Dorycnium graeeum (L.) Ser. 0.0±0.0 >1,000 >1,000
Dracocephalum moldavicum L. 93.3±5.5 789 (765–812) >1,000 3.189
Echinacea purpurea (L.) Moench 100.0±0.0 278 (250–308) 503 (440–598) 3.973
Echinops sphaerocephalus L. 38.3±6.3 >1,000 >1,000
Ferula assa-foetida L. 28.3±8.1 >1,000 >1,000
Galium verticillatum Danth. 0.0±0.0 >1,000 >1,000
Gilia capitata Sims 78.3±6.9 687 (620–732) >1,000 3.578
Grindelia camporum Greene 0.0±0.0 >1,000 >1,000
Hypericum perforatum L. 92.3±12.5 496 (428–512) 980 (923–997) 3.286
Imperatoria ostruthium L. 100.0±0.0 49 (44–54) 83 (74–101) 0.048
Inula auriculata Boiss. & Balansa 0.0±0.0 >1,000 >1,000
Jacobaea maritima (L.) Pelser & Meijden 100.0±0.0 270 (96–320) 428 (396–465) 3.745
Lapasana intermedia Bieb. 5.2±1.2 >1,000 >1,000
Lobelia siphilitica L. 100.0±0.0 633 (586–671) 929 (896–998) 4.126
Lythrum salicaria L. 30.0±5.3 >1,000 >1,000
Melilotus albus Medik. 0.0±0.0 >1,000 >1,000
Onobrychis miniata Stev. 8.6±3.4 >1,000 >1,000
Onopordon acanthium L. 1.7±0.5 >1,000 >1,000
Physalis alkekengi L. 91.7±9.3 899 (850–928) >1,000 3.112
Polygala comosa Schk. 0.0±0.0 >1,000 >1,000
Potentila erecta L. 0.0±0.0 >1,000 >1,000
Psoralea bituminosa L. 0.0±0.0 >1,000 >1,000
Pyrethrum corymbosum (L.) Willd. 10.0±2.5 >1,000 >1,000
Reseda odorata L. 100.0±0.0 381 (289–406) 915 (900–941) 1.386
Reynoutria × bohemica Chrtek & Chrtková 0.0±0.0 >1,000 >1,000
Reynoutria sachalinensis F. Schmidt 6.7±3.2 >1,000 >1,000
Rubia tinctorum L. 91.7±12.3 725 (712–762) 943 (932–1,002) 1.408
Salvia glutinosa L. 3.3±5.2 >1,000 >1,000
Saponaria officinalis L. (stem) 98.3±5.6 516 (498–552) 928 (895–998) 2.215
Saponaria officinalis L. (rhizome) 48.5±6.2 >1,000 >1,000
Scrophularia nodosa L. 75.3±8.2 982 (932–996) >1,000 1.863
Sedum rosea (L.) Scop. 0.0±0.0 >1,000 >1,000
Seseli pallasii Besser 100.0±0.0 6 (6–7) 10 (9–11) 0.065
Parasitol Res (2009) 105:887–892 891

Table 2 (continued)

Species Mortality % (mean±SE)a LD50b (CI95) LD90b (CI95) Chic

Schisandra chinensis (Turcz.) Baill 100.0±0.0 15 (13–17) 33 (28–39) 4.109

Silphium perfoliatum L. 0.0±0.0 >1,000 >1,000
Stachys byzantina K.Koch 100.0±0.0 65 (59–72) 133 (111–180) 3.565
Symphytum officinale L. 0.0±0.0 >1,000 >1,000
Teucrium hircanicum L. 100.0±0.0 316 (298–352) 688 (695–708) 1.285
Teucrium chamaedrys L. 100.0±0.0 710 (654–755) 957 (891–1,007) 2.858
Thymus fragrantissimus Samen. 10.5±2.3 >1,000 >1,000
Thymus serphyllum L. 100.0±0.0 711 (595–795) >1,000 1.023
Trifolium arvense L. 0.0±0.0 >1,000 >1,000
Tropaeolum majus L. 90.0±7.8 780 (695–898) >1,000 1.734
Verbena hirta Spreng. 10.0±2.3 >1,000 >1,000
Verbena officinalis L. 100.0±0.0 38 (28–46) 115 (100–141) 0.094
Vincetoxicum hirundinaria Medik. 100.0±0.0 717 (651–743) 957 (911–998) 3.860
Xeranthemum cylindraceum Smith. 8.3±3.4 >1,000 >1,000
a
Larvicidal activity (mean mortality) of plant extracts against fourth instar larvae of C. quinquefasciatus after 24 h of exposure and maximal dose
1,000 ppm
b
The lethal concentration with the corresponding 95% confidence intervals are shown in parenthesis
c
Chi-square for heterogeneity (calculated)

Results and discussion development of botanical larvicides. The good efficacy of

plant extracts, or substances obtained from plants, on the
The results of the larvicidal activity of plant extracts are larvae mortality of mosquitoes has been found by other
presented in Table 2. Forty-nine plants showed larvicidal authors as well. Treatment of stagnant water with larvicides
activity in 24-h exposure tests at the maximum tested dose is generally regarded as a very good method of reducing the
(1,000 ppm). The methanolic extracts of the plants Seseli total number of adult mosquitoes. Our results and the works
pallasii and Schisandra chinencis displayed the highest of other authors indicate great potential for using plant
larvicidal activities with LD50 6 and 15 ppm, respectively, extracts as larvicides. Botanical larvicides can contribute
followed by Arthemisia campestris, Verbena officinalis, and remarkably to lowering the vector population of mosquitoes
Imperatoria osthruthium with LD50 23, 38, and 49 ppm, (Ascher and Meisner 1989; Pizzarro et al. 1999; Omena et
respectively. The appraised value of LD50 for two species al. 2007; Kamaraj et al. 2008; Rahuman et al. 2008).
was between 51–100 ppm; eight species had lethal doses The biological activity of these plant extracts might be
from 101 to 500 ppm; 13 species showed lethal doses from due to the various compounds, including phenolics,
501 to 1,000 ppm; and 34 species did not show lethal doses terpenoids, and alkaloids, that exist in plants; these
for low mortality (LD50 >1,000 ppm). compounds may contribute jointly or independently to
Today, the environmental safety of an insecticide is producing larvicidal and adult emergence inhibition activity
considered to be of paramount importance. The plants against C. quinquefasciatus. Further investigation is needed
tested for this and the former work (Pavela 2008) were to elucidate this activity against a wide range of all stages
predominantly medicinal plants, which highly presuppose of mosquito species. In addition, the active ingredients of
the absence of health damaging substances. For this reason, the extract responsible for larvicidal and adult emergence
it is necessary to test those plant extracts which could be inhibition activity in C. quinquefasciatus should be identi-
expected to contain substances of environmental potential, fied and used, if possible, in preparing a commercial
and at the same time, provide adequate efficacy against product/formulation for use as a mosquitocide.
mosquitoes (Trongtokit et al. 2008). This work has
identified five plants with lethal doses estimated at lower
than 50 ppm. Although the efficacy of the extracts on larvae Acknowledgements This study was supported by grants from the
Czech Republic Ministry of Education, Youth, and Sports (ME09079).
mortality has been studied by a lot of authors, such good The author is grateful to Prof. V. M. Kalinkin, Dr. T. Chermenskaya,
efficacy on mortality is infrequent. For this reason, these and Dr. A. Shchenikova from VIZR, St. Petersburg, Russia for help in
plants can be regarded as having strong prospects for the carrying out the expedition and the collection of plants.
892 Parasitol Res (2009) 105:887–892

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