RESUMO: &quot;Extratos de Os bioensaios mostraram inibição no desenvolvimento pós-embrionário das moscas e um signifi cativo aumento do período larva recém eclodida-adulto. Além disso, o peso pupal foi reduzido e a proporção sexual alterada. Foi observada toxicidade para os ovos das moscas. Unitermos: Extratos de planta, Diptera, Melia azedarach, regulação do desenvolvimento, atividade biológica. ABSTRACT: Crudes extracts and fractions from seeds of Melia azedarach L. (Meliaceae) have been assayed on Musca domestica Linnaeus, 1758 (Diptera: Muscidae). Thus, the postembryonic development of the fl ies was reduced and the delay from newly hatched larvae to adults had signifi cant increase. In addition, the pupal weights were reduced and the sexual ratio altered. Toxicity to fl y eggs was also observed

Alphonse Kelecom

Celma M S Gomes

Esteban R F Crescente

Marise M O Cabral

Paloma M Mendonça

Revista Brasileira De Farmacognosia

Vanderleia C Oliveira

CiteSeerX

699
A
rti
go
ISSN 0102-695X
Revista Brasileira de Farmacognosia
Brazilian Journal of Pharmacognosy
18 (Supl.): 699-702, Dez. 2008
* E-mail: mmaleck@oi.com.br, Tel. +55-21-25621213, Fax +55-21-25755968
Received 4 October 2008; November 5 2008
Melia azedarach L. extracts and their activity on Musca domestica 
L. (Diptera: Muscidae)
Marise M. O. Cabral,*,1,2 Esteban R. F. Crescente,4 Paloma M. Mendonça4 Celma M. S. 
Gomes,3 Vanderleia C. Oliveira,4 Alphonse Kelecom5
1Laboratório de Diptera, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Pavilhão Carlos 
Chagas, 413, 21045-900 Rio de Janeiro-RJ, Brazil,
2Laboratório de Insetos Vetores, Universidade Severino Sombra, Av. Expedicionário Oswaldo de Almeida Ramos, 
280, 27700-000 Vassouras-RJ, Brazil,
3Laboratório de Hematozoários, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Pavilhão 
Carlos Chagas, 4º andar, 21045-900 Rio de Janeiro-RJ, Brazil,
4Laboratório de Ecoepidemiologia da Doença de Chagas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. 
Brasil 4365, 21045-900 Rio de Janeiro-RJ, Brazil,
5Departamento de Biologia Geral, Universidade Federal Fluminense, Outeiro de São João Batista s/n, Cx. Postal 
100.436, 24001-970 Niterói-RJ, Brazil
RESUMO: “Extratos de Melia azedarach L. e sua atividade sobre Musca domestica L. 
(Diptera: Muscidae)”. Os extratos brutos e as frações obtidas das sementes de Melia azedarach 
L. (Meliaceae) foram testados em Musca domestica Linnaeus, 1758 (Diptera: Muscidae). 
Os bioensaios mostraram inibição no desenvolvimento pós-embrionário das moscas e um 
signifi cativo aumento do período larva recém eclodida- adulto. Além disso, o peso pupal foi 
reduzido e a proporção sexual alterada. Foi observada toxicidade para os ovos das moscas.
Unitermos: Extratos de planta, Diptera, Melia azedarach, regulação do desenvolvimento, 
atividade biológica. 
ABSTRACT: Crudes extracts and fractions from seeds of Melia azedarach L. (Meliaceae) 
have been assayed on Musca domestica Linnaeus, 1758 (Diptera: Muscidae). Thus, the post-
embryonic development of the fl ies was reduced and the delay from newly hatched larvae to 
adults had signifi cant increase. In addition, the pupal weights were reduced and the sexual ratio 
altered. Toxicity to fl y eggs was also observed.
Keywords: Plant extract, Diptera, Melia azedarach, growth development, biological activity.
INTRODUCTION
The Meliaceae Melia azedarach L. is a huge 
tree, natural from South Asia and very common in 
Brazil, where it is known as “Cinamomo”. A number 
of biological activities have already been described for 
crude extracts, fractions and metabolites isolated from 
this plant such as giardicial activity (Amaral et al., 
2006), inhibitors of the enzyme acetylcholinesterase 
(Barbosa-Filho et al., 2006a), inhibitors of the 
angiotensin converting enzyme (Barbosa-Filho et al., 
2006b) and antileprotic activity (Barbosa-Filho et al., 
2007). Antitumor (Hartwell, 1971), anti-rheumatic, 
sedative and antiulcer properties (Duke & Wain, 1981; 
List & Horhammer, 1979) and used against fl u and 
hypertension (Agra et al., 2008) are reported in traditional 
medicine. The plant also possesses insecticide (Cabral 
et al., 1995; Lepage et al., 1946) and vermicide activities 
(Zhao, 1984). Among secondary metabolites found in 
M. azedarach, one should mention triterpenes (Cabral 
et al., 1996; Mulholland et al., 2000), steroids, aromatic 
compounds (Mulholland et al., 2000) and the interesting 
limonoid meliaternin (Carpinella et al., 2003). In 
previous studies, we described a new euphane triterpene 
(Kelecom et al., 1996) and four lignans (Cabral et al., 
1995) from the methanol extract of the seeds of M. 
azedarach collected in the city of Niterói, RJ. This was 
the fi rst report of lignans in the family Meliaceae. We 
also reported anti-moulting activity of this crude extract 
against the bloodsucking bug Rhodnius prolixus (Cabral 
et al., 1996). The study described the biological activity 
of extracts and fractions of M. azedarach L. on the post-
embryonic development of Musca domestica Linnaeus, 
1758 (Diptera: Muscidae). 
MATERIAL AND METHODS
Seeds of M. azedarach were dried, powdered 
Revista Farmacognosia Suplemento 2008.indd   Sec1:699 23/01/2009   10:07:05
700
Marise M. O. Cabral, Esteban R. F. Crescente, Paloma M. Mendonça Celma M. S. Gomes, Vanderleia C. Oliveira, Alphonse Kelecom
Rev. Bras. Farmacogn.
Braz J. Pharmacogn.
18 (Supl.): Dez. 2008
and successively extracted with n-hexane, AcOEt (A) 
and MeOH (B). The oily hexane crude extract was 
then partitioned between cold hexane and 5% aqueous 
methanol to yield a hexane fraction (C) and a methanol 
fraction (D). Aliquots of the extracts A and B, and of the 
fractions C and D were carefully dried, traces of organic 
solvents eliminated under vacuum, and the organic 
residues bioassayed on M. domestica, a cosmopolitan 
diptera with high degree of synanthropy that acts as a 
mechanical and biological vector for enteroviruses, 
enterobacteria, protozoan cysts, helminth eggs and 
larvae, and fungi (Cox et al., 1912, Coutinho et al., 
1957, Greenberg, 1971, 1973, Harwood & James, 1979; 
Oliveira, 1999; Oliveira et al., 2002). Insects used in 
this study were brought up in the colony maintained in 
laboratory (Queiroz et al., 1991). Samples were tested 
in triplicate by topic application at the concentration of 
100 μg/μL on groups of 50 larvae (L1) of M. domestica 
and on egg masses (5 mg) at the concentration of 200 
μg/mg (1 μL/mg). Insect viability, duration periods of 
larval, pupal and newly hatched larvae to the adult stage, 
were observed as well as sexual ratio and pupal weight. 
Treatment 
Larval stage 
viability 
(%) 
Pupal stage 
viability 
(%) 
Larvae to adult 
viability 
(%) 
Sex ratio 
Control 85 a 85 
a 
wt (mg) = 22.10 ±3.90 a 73 
a 0.44 
A 79 c *** 68 
b ** 
wt (mg) = 20.17 ±3.28 b*** 53 
b ** 0.50 
B 77 c *** 65 
b * 
wt (mg) =20.88 ±3.31 a 50 
b *** 0.55 
C 73 c *** 68 
b *** 
wt (mg) =21.52 ±3.47X a, c* 50 
b *** 0.49 
D 54 b *** 69 
b ** 
wt (mg) = 20.41 ±3.69 b, c** 37 
c *** 0.38 
Treatment Larval stage (days)  
Pupal stage 
(days)  
Larvae to adult 
(days)  
 X ± SD VI X ± SD VI X ± SD VI 
Control 6.58 ± 0.77 a 5-10 5.56 ± 0.69 a 2-7 12.08 ± 0.62 a 11-14 
A 6.45 ± 0.66 a 5 -8 5.89 ± 0.52 b** 4-8 12.25 ± 0.68 a 10-14 
B 6.43 ± 0.76 a* 5-10 5.96 ± 0.52 b*** 5-7 12.35 ± 0.86 a 11-14 
C 6.72 ± 0.81 b* 5-10 5.96 ± 0.64 b*** 5-8 12.63 ± 0.83 b*** 11-14 
D 6.65 ± 0.74 a 5-10 5.98 ± 0.65 b*** 2-7 12.59 ± 0.65 b*** 11-14 
Treatment Number of eggs Number of larvae Egg viability (%) 
Control 167 152 91 a 
Acetone control 266 233 88 b * 
Fraction D 245 54 22 c *** 
Table 1. Viability (%) and sex ratio of post-embryonic development of M. domestica treated with extracts from M. azedarach, topic 
application on L1 larvae.
Triplicate experiments with groups of 50 larvae of M. domestica, each. Numbers followed by the same letter did not differ among 
themselves and those followed by different letters have a signifi cant difference (* P < 0.05, ** P < 0.01, ***P < 0.001) when the 
Tukey test was used; wt  = pupal weight (mg), values are mean ± standard deviation (X ± SD); sex ratio refer to ratio number of 
females versus number of males.
Table 2. Duration in days of post-embryonic development of M. domestica treated with extracts from M. azedarach, topic 
application on L1 larvae.
Triplicate experiments with groups of 50 larvae of M. domestica, each. VI= variation interval. Means followed by the same letter 
did not differ among themselves and those followed by different letters have a signifi cant difference (* P < 0.05, ** P < 0.01, ***P 
< 0.001), when the Tukey test was used. Values are mean ± standard deviation (X ± SD). 
Table 3. Viability of eggs (%) of M. domestica treated with fraction D from M. azedarach, topic application on eggs mass at the 
concentration of 200 μg/mg.
Triplicate experiments with groups of eggs mass (5mg) of M. domestica, each.  Numbers followed by the same letter did not differ 
among themselves and those followed by different letters have a signifi cant difference (* P < 0.05, ***P < 0.001) when the test χ2 
was used.
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701
Melia azedarach L. extracts and their activity on Musca domestica L. (Diptera: Muscidae)
Rev. Bras. Farmacogn.
Braz J. Pharmacogn.
18 (Supl.): Dez. 2008
The results were expressed by their means and standard 
deviations, and the signifi cance established statistically 
using the χ2 and the Tukey tests.
RESULTS
The results are summarized in Tables 1-3. Thus, 
the post-embryonic development of the fl ies appeared 
to be drastically reduced on treatment with samples 
B, C and D, showing a viabilility of 50, 50 and 37% 
for each sample, respectively (Table 1). The viability 
of the larval stage was reduced by all tested samples 
of M. azedarach, mainly by fraction D that induced 
a reduction of 31% when compared with those of the 
control groups.  The pupal weight was reduced by all 
the samples as compared to the control group (22 mg) 
with concomitant reduction of the fl ies size; thus, mean 
weights of 20 mg (spreading: 10-28 mg) were observed 
when larvae were treated with A (P < 0.001) and D (P < 
0.01) respectively (Table 1). In addition, the sexual ratio 
(number females/number males) showed a reduction of 
the number of females (ratio = 0.38) in the group treated 
with fraction D (Table 1).
The duration of the pupal stage showed a 
signifi cant increase when M. domestica were treated with 
samples A (P < 0.01), B (P < 0.001), C (P < 0.001) and 
D (P < 0.001). The duration of the newly hatched larvae 
to adults also had signifi cant increase when treated with 
samples C (P < 0.001) and D (P < 0.001) (Table 2). 
Treatment of egg mass with D, at the 
concentration of 200 μg/mg egg mass, resulted in only 
22% of hatched larvae (Table 3) thus proving the high 
toxicity of this fraction from M. azedarach seeds on M. 
domestica eggs.
DISCUSSION AND CONCLUSION
This study describes preliminary results of 
the analysis of the biological activity of M. azedarach 
extracts focusing toxicity, pupation inhibition and 
outbreak of M. domestica, looking for fractions and 
substances that might be effi cient to control dipterans 
vectors of pathogens. Inhibition of the post-embryonic 
development of M. domestica was observed in extract D 
that induced 46% and 63% larval and outbreak inhibitions 
respectively. Some authors described that an extract of 
the seeds of this tree is able to exert a repellent activity 
on Locusta migratoria (Pradhan et al., 1962), and, that 
the acetone extract interferes on the pupal development 
of Culex pipiens (Al Sharook et al., 1991). Similarly, 
anti-molting activity was found in M. azedarach against 
the triatomine insect R. prolixus, vector of Chagas’s 
disease (Cabral et al., 1995, 1996). 
High toxicity against larvae and pupae of M. 
domestica was evidenced in fractions C and D, both 
obtained by partition of the hexane crude extract from the 
Meliaceae. Moderate to high mortality of M. domestica 
was observed when these fl ies were exposed to extracts 
from Gymnospermae from New Zeeland (Singh & 
Upadhyay, 1993). A similar result was found for the 
AcOEt (80%) and hexane (85%) crude extracts on R. 
prolixus (Cabral et al., 1996). The same authors assayed 
extracts of M. azedarach and demonstrated molting 
inhibition and toxicity on the Hemiptera O. fasciatus and 
on the hematophagous triatomine R. prolixus (Cabral et 
al., 1999). Purifi cations of the crude extracts increased 
the activities. On fractionation, the anti-molting activity 
was found in fraction B, that inhibited 80% of the 
ecdysis of R. prolixus (Cabral et al., 1996), the active 
principle being identifi ed as pinoresinol, that showed 
90% (25 μg/μL) and 65% (25 μg/μL) molting inhibition 
on O. fasciatus and R. prolixus, respectively (Cabral et 
al., 1999).
Studies on species of Meliaceae of the genus 
Trichilia also appeared promising for insecticide use 
on soil plagues and on the armyworm Spodoptera 
frugiperda (Hernández et al., 1983). Similarly in the 
health area, several vegetal extracts are under study 
aiming at the control of insects, vectors of illnesses 
to man, such as hematophagous mosquitoes, domestic 
fl ies, cockroaches and earwigs (Lagunes et al., 1984, 
Simas et al., 2004).
In conclusion, the seed extracts of M. azedarach 
showed potent bioactivity against M. domestica, which 
is an important urban pest all over the world. Sample 
D demonstrated the highest activity. This seems to be 
the fi rst report of biological activities of M. azedarach 
extracts against Muscidae insects.
ACKNOWLEDGEMENTS
This work was supported by grants from the 
Fundação de Amparo à Pesquisa do Estado do Rio de 
Janeiro (FAPERJ).
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Melia azedarach L. extracts and their activity on Musca domestica L. (Diptera: Muscidae)

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Melia azedarach L. extracts and their activity on Musca domestica L. (Diptera: Muscidae)

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