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

Extratos de Melia azedarach L. e sua atividade sobre Musca domestica L. (Diptera: Muscidae)

ARTIGO

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

Extratos de Melia azedarach L. e sua atividade sobre Musca domestica L. (Diptera: Muscidae)

Marise M. O. Cabral^{I, II, *}; Esteban R. F. Crescente^IV; Paloma M. Mendonça^IV; Celma M. S. Gomes^III; Vanderleia C. Oliveira^IV; Alphonse Kelecom^V

^ILaboratório de Diptera, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Pavilhão Carlos Chagas, 413, 21045-900 Rio de Janeiro-RJ, Brazil

^IILaboratório de Insetos Vetores, Universidade Severino Sombra, Av. Expedicionário Oswaldo de Almeida Ramos, 280, 27700-000 Vassouras-RJ, Brazil

^IIILaboratório de Hematozoários, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Pavilhão Carlos Chagas, 4º andar, 21045-900 Rio de Janeiro-RJ, Brazil

^IVLaboratório de Ecoepidemiologia da Doença de Chagas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, 21045-900 Rio de Janeiro-RJ, Brazil

^VDepartamento de Biologia Geral, Universidade Federal Fluminense, Outeiro de São João Batista s/n, Cx. Postal 100.436, 24001-970 Niterói-RJ, Brazil

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 flies was reduced and the delay from newly hatched larvae to adults had significant increase. In addition, the pupal weights were reduced and the sexual ratio altered. Toxicity to fly eggs was also observed.

Keywords: Plant extract, Diptera, Melia azedarach, growth development, biological activity.

RESUMO

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 significativo 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.

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 flu 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 first 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 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. The results were expressed by their means and standard deviations, and the significance 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 flies 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 flies 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 significant 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 significant 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 efficient 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 flies 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). Purifications 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 identified 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 flies, 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 first 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).

Received 4 October 2008

November 5 2008

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Publication Dates

• Publication in this collection
  09 Mar 2009
• Date of issue
  Dec 2008

History

• Accepted
  05 Nov 2008
• Received
  04 Oct 2008