VECTOR CONTROL, PEST MANAGEMENT, RESISTANCE, REPELLENTS
Disruption of Chrysomya megacephala Growth Caused by
Lignan Grandisin
CAMILA DINIZ RIBEIRO NOGUEIRA,1 RUBENS PINTO DE MELLO,1 MASSUO JORGE KATO,2
1,3,4
AND MARISE MALECK DE OLIVEIRA CABRAL
J. Med. Entomol. 46(2): 281Ð283 (2009)
KEY WORDS Chrysomya megacephala, grandisin, lignan, Diptera, postembryonic development
Piper (Piperales: Piperaceae) species are rich source
of bioactive compounds, including amides, chromenes,
and lignoids (Sengupta and Ray 1987, Jensen et al. 1993,
Benevides et al. 1999, Lago et al. 2004). The tetrahydrofuran grandisin has been isolated as the major compound from Piper solmsianum C.DC., an endemic species from Mata Atlântica in Brazil, and it has displayed
powerful trypanocidal activity (Martins et al. 2000,
Kato and Furlan 2007).
Chrysomya megacephala F. (Diptera: Calliphoridae) originally from Africa was introduced into southern Brazil during 1975Ð1976 (Guimarães et al. 1978).
Due to feeding and reproductive traits, it has important role as vector for several pathogens and diseases
(Greenberg 1973, Oliveira et al. 2002). C. megacephala
also has the ability to produce primary and secondary
myiasis to animals, including humans (Sukontason et
al. 2005). Herein, the toxicity of grandisin on C. megacephala larvae, including postembryonic development, is described.
Materials and Methods
Grandisin Isolation. (⫺)-Grandisin was extracted
from dried inßorescences of P. solmsianum as described by Martins et al. (2003), collected from a
garden at the Institute of Chemistry, USP, São Paulo,
Brazil, and its identiÞcation has been reported previ1 Laboratório de Diptera/Entomologia/IOC, Fundação Oswaldo
Cruz, Av. Brasil 4365, 21040-900 Rio de Janeiro, RJ, Brazil.
2 Departamento de Quṍmica Fundamental/Instituto de Quṍmica,
Universidade de São Paulo, USP, Av. Prof. Lineu Prestes 748, 05508900 São Paulo, SP, Brazil.
3 Departamento de Ciências Biológicas, Universidade Severino
Sombra, Av. Expedicionário Oswaldo de Almeida Ramos 280, 27700000 Vassouras, RJ, Brazil.
4 Corresponding author, e-mail: mmaleck@ioc.Þocruz.br.
ously (Martins et al. 2000). It was dissolved in acetone
and diluted at 1:4 in 0.8% NaCl to a concentration of
100 ␮g/␮l.
Flies. Adult ßies collected at Seropédica, Rio de
Janeiro, maintained the colony at the Laboratory of
Vector Insect Biology and Control, IOC, FIOCRUZ,
RJ, Brazil, as described previously (Queiroz and Milward-de-Azevedo 1991).
Topical Treatment of Egg Masses. Grandisin solution was applied with a pipette to the egg masses (1
␮l/mg eggs). Two control groups were used: one
group received an application of only acetone solution, and one group of egg masses received no treatment.
Topical Treatment of Larvae and Starved Larvae.
Grandisin solution was applied to Þrst-instar larval
bodies (F1ÐF5) (1 ␮l per larva). In starved larvae,
treatment was carried out immediately after the larvae
hatched, and no meat was offered to any of the groups
of Þrst instars, except for the feeding control. The
larval test group was fed 45 min after treatment. Three
control groups were used: larvae with application of
diluted acetone solution (without grandisin and
food), larvae without treatment and food (starved
control), and larvae untreated with normal diet (feeding control).
Feeding Larvae with a Treated Diet. Grandisin solution was applied to an artiÞcial diet (Mendonça and
dÕAlmeida 2004) of dried whole milk (1 g per larvae
and 1 ␮l/mg diet). Two control groups were used:
larvae feeding on artiÞcial diet only with acetone solution and larvae feeding on untreated artiÞcial diet.
Groups of larvae receiving topical (egg masses and
larvae) treatments were fed on a diet of putreÞed
bovine meat (1 g per larva). Mature larvae were individually weighed on a precision balance and trans-
0022-2585/09/0281Ð0283$04.00/0 䉷 2009 Entomological Society of America
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ABSTRACT The toxicity of tetrahydrofuran lignan grandisin was evaluated against larvae of Chrysomya megacephala F. (Diptera: Calliphoridae). The bioassay involved topical treatment on larvae,
topical treatment on egg masses, and incorporation in the larval diet. Grandisin showed inhibition of
postembryonic development by ovicidal (30%) and larvicidal (38%) effects and reduced larval weight
(4 mg), when topically applied on egg masses and starving larvae (L1) at a concentration of 100 ␮g/␮l.
These Þndings elucidated the effect of grandisin on the C. megacephala life cycle and its potential to
control C. megacephala populations.
282
JOURNAL OF MEDICAL ENTOMOLOGY
Table 1.
Vol. 46, no. 2
Viability (percentage) of postembryonic development of C. megacephala treated with grandisin (100 ␮g/␮l)
Treatment
1.1. Topical on eggs
Control
Acetone control
Grandisin
1.2. Topical on larvae
Control
Acetone control
Grandisin
1.3. Treatment in diet
Control
Acetone control
Grandisin
1.4. Topical on starved larvae
Feeding control
Starved control
Acetone control
Grandisin
Eggs
Larvae
Pupae
Adults
Mean ⫾ SD
%
Mean ⫾ SD
%
Mean ⫾ SD
%
Mean ⫾ SD
%
70.7 ⫾ 3.8
67.7 ⫾ 11.2
39.0 ⫾ 19.5
81a
78a
45*b
57.3 ⫾ 3.8
58.7 ⫾ 8.7
32.3 ⫾ 18
81a
87a
83a
57.3 ⫾ 3.8
58.3 ⫾ 8.7
32.3 ⫾ 18
100a
99a
100a
52.3 ⫾ 5.7
56.0 ⫾ 7.9
29.3 ⫾ 19.7
91a
96a
91a
26.3 ⫾ 2.0
19.5 ⫾ 2.1
16.3 ⫾ 4.5
88a
65ab
54ⴙb
26.3 ⫾ 2.0
19.5 ⫾ 2.1
16.3 ⫾ 4.5
100a
100a
100a
26.0 ⫾ 1.7
19.5 ⫾ 2.1
15.3 ⫾ 5.5
99a
100a
94a
20.0 ⫾ 2.8
14.6 ⫾ 2.0
13.0 ⫾ 2.7
67a
49a
43a
20.0 ⫾ 2.8
14.6 ⫾ 2.0
13.0 ⫾ 2.7
100a
100a
100a
18.5 ⫾ 2.1
14.0 ⫾ 3.0
13.0 ⫾ 2.7
93a
95a
100a
24.3 ⫾ 0.6
22.3 ⫾ 3.1
20.6 ⫾ 1.5
12.7 ⫾ 6.8
97a
89a
83a
51*b
24.0 ⫾ 1.0
22.3 ⫾ 3.1
20.6 ⫾ 1.5
12.7 ⫾ 6.8
99a
100a
100a
100a
22.7 ⫾ 0.6
20.3 ⫾ 4.6
19.0 ⫾ 2.0
11.7 ⫾ 6.5
94a
91a
92a
92a
ferred to glass tubes containing vermiculite (one
fourth) for emergence of ßies.
The bioassays were performed in triplicate, repeated on three occasions with different batches of
insects, each with three repetitions maintained in a
climate chamber without light control. Ten adult ßies
were sexed, and the left tibia was measured under a
stereomicroscope to estimate size. The estimated
weight, size, viability, and sexual ratio were recorded
daily during each phase of development. The data
were analyzed by Tukey test (5% signiÞcance level)
(InSTAT; GraphPad Software Inc. 1999).
Results and Discussion
Egg masses receiving grandisin treatment showed
signiÞcant increases in duration of the larval stage
(5.67 ⫾ 0.59) (P ⬍ 0.001) compared with larvae from
the acetone control group (5.17 ⫾ 0.44). When C.
megacephala starved larvae were topically treated
with grandisin, the durations of larval stage and period
from newly hatched larva to adult were 4.92 ⫾ 0.36 and
9.71 ⫾ 0.51 d, respectively. These time lengths were
signiÞcantly different (P ⬍ 0.001) from those of the
acetone control (4.57 ⫾ 0.53 and 9.25 ⫾ 0.43).
The eggs and the whole period from egg to adult
seemed to be particularly sensitive to grandisin, with
viability of only 45% (P ⬍ 0.1) (Table 1, 1.1), causing
ultimately 33% lower egg eclosion. The larval stage
also seemed to be particularly sensitive to topical
treatment with grandisin, showing larval viability of
54% (Table 1, 1.2). Diet treatment with grandisin and
acetone decreased the viability of the larvae by ⬇20%
(Table 1, 1.3). Topical treatment on starved larvae
showed 51% larval stage viability (P ⬍ 0.1) 32% mortality (Table 1, 1.4). The topical grandisin treatment
on eggs caused a decrease of 4 mg in larval weight
(68.21 ⫾ 11.41) (P ⬍ 0.01) and 0.15 mm in size of ßies
(2.71 ⫾ 0.07) (P ⬍ 0.1), compared with acetone control (72.13 ⫾ 7.97 and 2.86 ⫾ 0.1, respectively).
Grandisin treatment caused inhibition of growth
and development of C. megacephala, through larvicidal and ovicidal activities. Cabral et al. (2007a,b)
showed that the viability of larvae at similar stages
decreased 60% in C. megacephala treated with
yangambin, burchellin neolignans, or both. These results demonstrate that lignoids with different skeletal
types lead to selective disruption in development of
dipterans (Chauret et al. 1996, Solis et al. 2005).
A possible source to consider for grandisin should
be P. solmsianum in which the content is substantial
(nearly 5% in dry weight mass). Although still growing
in the wild, it is not difÞcult to cultivate P. solmsianum.
In addition, grandisin can be obtained directly by
crystallization from crude ethanol extracts. Further
studies are required to elucidate the mechanism of
action for grandisin as a natural insecticide product for
uses in blowßy population control.
Acknowledgments
We are indebted to Chagas Disease Eco-epidemiology
Laboratory, FIOCRUZ, and Colégio Pedro II for technical
assistance with the insect colony. This research was supported by grants from Fundação de Amparo à Pesquisa do
Estado do Rio de Janeiro (FAPERJ) and Coordenação de
Aperfeiçoamento de Pessoal de Nṍvel Superior.
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