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Effect of Used Coffee Grounds on Larval Mortality of Aedes aegypti L. (Diptera: Culicidae): Suspension Concentration and Age versus Efficacy

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In a previous study, used coffee ground affected the larval development of Aedes aegypti. In this work, we evaluated the duration of the effect on larval mortality of aqueous suspensions of used coffee ground at 75, 150, 250 and 300 mg/ml concentrations. The larval mortality was followed daily, in the experimental breeding sites; 300 mg/ml was the most efficient concentration, producing 100% of larval mortality until nine days after preparation. These results were observed in experiments in which the larvae remained free in the vials, making contact both the used coffee ground deposits and the supernatant liquid, and also in experiments in which the larvae were maintained in a sieve of fine screen immersed in the liquid part of the suspension. Thus, although the elimination of the breeding sites remains being the best way to control Aedes aegypti population size, the results obtained herein reinforces the validity of considering used coffee ground preparations as possible auxiliary in the alternative control of this mosquito, mainly in the gardens. Used coffee ground has the advantage of being free of cost, since it is the powder that is left after coffee has been filtered out to drink
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Content Preview
Received: 06/X/2006
Accepted: 10/IV2007
Published: 31/V/2007

Available online at: www.bioassay.org.br/articles/2.5





CHEMICAL CONTROL

Effect of Used Coffee Grounds on Larval Mortality of Aedes aegypti
L. (Diptera: Culicidae): Suspension Concentration and Age versus
Efficacy

MARLUCI MONTEIRO GUIRADO AND HERMIONE ELLY MELARA DE CAMPOS BICUDO

Universidade Estadual Paulista (UNESP), Instituto de Biociências, Letras e Ciências Exatas (IBILCE),
Departamento de Biologia. Rua Cristóvão Colombo, 2265, Jardim Nazareth, CEP: 15054-000, São José do Rio
Preto, SP, Brazil. E-mail: bicudo@ibilce.unesp.br.

BioAssay 2:5 (2007)

Efeito da Borra de Café na Mortalidade Larval de Aedes aegypti L. (Diptera: Culicidae): Concentração e
Idade da Suspensão versus Eficácia

RESUMO –
Em estudo anterior, a borra do café afetou o desenvolvimento larval de Aedes aegypti L. No presente
trabalho, foi analisada a duração do efeito na mortalidade larval, de suspensões aquosas de borra do café, nas
concentrações 75, 150, 250 e 300 mg/ml. A mortalidade larval nos criadouros experimentais foi acompanhada
diariamente: a concentração 300 mg/ml foi a mais eficiente, produzindo 100% de mortalidade até nove dias após o
preparo da suspensão. Estes resultados foram observados tanto em experimentos nos quais as larvas permaneceram
livres podendo fazer contato com o depósito de borra do café no fundo dos frascos e com o sobrenadante líquido,
como também nos experimentos em que as larvas foram mantidas em uma peneira de tela fina imersa na parte
líquida da suspensão. Embora a eliminação dos criadouros seja a melhor maneira de controlar o tamanho das
populações de A. aegypti, os resultados deste estudo reforçam a validade de considerar a borra do café como um
possível auxiliar no controle deste mosquito, principalmente em jardins. A borra do café tem a vantagem de ser livre
de custo, pois é o pó deixado no coador e jogado fora depois que a bebida é preparada.

PALAVRAS-CHAVE: Aedes aegypti, controle alternativo, borra do café

ABSTRACT - In a previous study, used coffee ground affected the larval development of Aedes aegypti. In this
work, we evaluated the duration of the effect on larval mortality of aqueous suspensions of used coffee ground at 75,
150, 250 and 300 mg/ml concentrations. The larval mortality was followed daily, in the experimental breeding sites;
300 mg/ml was the most efficient concentration, producing 100% of larval mortality until nine days after
preparation. These results were observed in experiments in which the larvae remained free in the vials, making
contact both the used coffee ground deposits and the supernatant liquid, and also in experiments in which the larvae
were maintained in a sieve of fine screen immersed in the liquid part of the suspension. Thus, although the
elimination of the breeding sites remains being the best way to control Aedes aegypti population size, the results
obtained herein reinforces the validity of considering used coffee ground preparations as possible auxiliary in the
alternative control of this mosquito, mainly in the gardens. Used coffee ground has the advantage of being free of
cost, since it is the powder that is left after coffee has been filtered out to drink.

KEYWORDS: Aedes aegypti, alternative control, used coffee ground


The mosquito Aedes aegypti L. is presently
presently by human activities, part of them potential
considered the one that shows the greatest dispersion in
breeding sites for the mosquito and the globalization of
urban areas of the world (Silva et al. 2004). In many of
human activities allowing the transport of mosquitoes
those regions, A. aegypti is a vector of virus that causes
between regions and countries. The mosquito is
serious human diseases, such as dengue, dengue
controlled mainly by the use of insecticides, whose
hemorrhagic fever and yellow fever. Several factors
toxic effects for man and the environment are well
contribute for the high density of mosquito populations.
known (Slosek 1986, Marzochi 1994, Chauhan et al.
Among them are the great amount of garbage produced
2000, Tian et al. 2000). The situation is being



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1




Effect of coffee grounds on Aedes aegypti
Guirado e Bicudo

aggravated by the fact that the mosquitoes are
150, 225 and 300 mg/ml, corresponding approximately
developing insecticide-resistance, causing the frequent
to one, two, three or four level tablespoons,
need for the application of higher doses of those
respectively. Five glasses were prepared of each of the
substances, their improper use in mixtures and the
four concentrations and of the tap water used as control.
substitution of one insecticide by another of stronger
Daily, 10 larvae were placed into five glasses, four of
effects (Macoris et al. 2003, Sousa-Polezzi & Bicudo
which contained a different concentration of the
2004). These facts make it highly desirable to find
medium and the fifth the control, till all the replicates
alternative ways to control the Aedes population size.
had been used. The glasses were covered with a fine
Mainly products originate from plants have been
mesh fabric fixed with a rubber band. The analysis was
described for this aim (Furtado et al. 2005; Promsiri et
daily, approximately in the same schedule, computing
al. 2006).
the number of live and dead larvae and the range
Used coffee grounds (UCG), the powder that is
treatment time to death. When all the larvae in a glass
left after coffee has been filtered out to drink was used
had died or developed to the adult stage, it was reused
in tests for mortality of A. aegypti (Laranja et al. 2003).
for 10 new larvae. The analyses in this experiment
UCG suspensions in appropriate concentrations
involved suspensions aged up to 12 days, due to this
blocked the development of the mosquito before
reuse of the glasses. The age is the time elapsed since
reaching the adult phase, in which the virus
the suspension was prepared. Fish food was not added
transmission occurs through the bites of the female.
to the suspensions.
In the present study, experiments for increasing
Second Experiment. The larvae were maintained in a
information referring to the use of UCG were carried
sieve of fine screen immersed into the liquid part of the
out, looking at the possibility of its use as auxiliary in
UCG suspension, restricting them from touching or
the alternative control of A. aegypti. More specifically,
going into the UGC deposit located at the bottom of
in this study the durability of the effect of UCG
the treatment glass. This experiment involved the same
suspensions on larval mortality (LM) was analyzed,
concentrations and the same number of replicates of
since this is an important aspect of any substance-
the First Experiment. Two of the five replicates in each
candidate for the aforesaid purpose.
concentration (the one-day and four-day suspensions)

received the addition of 0.01 g of food per 200 ml of
Material and Methods
suspension. The analysis was carried out as in the First
Experiment, and suspensions aged up to 15 days were
In this study, larvae and pupae of A. aegypti were
used.
collected two or three times a month in breeding sites
(rain-filled tires, cans, etc.), in an urban area from São
Statistical Analysis
Statistical analysis involved the use of linear
José do Rio Preto - SP, by technicians from SUCEN
correlation between UCG suspension age and
and brought to the Vector Laboratory, at the
percentage larval mortality in 48h (Zar, 1999).
Department of Biology – UNESP / IBILCE, where

they were used to originate the cultures. In the
laboratory, the mosquitoes, in the above-mentioned
Results and Discussion
stages were placed for development in glasses
First Experiment. The results of this experiment,
(tumblers) containing tap water, inside wood cages
which involved the use of larvae placed unrestrictedly
with fine nylon screen walls. Their progeny, in the
in the medium, are set out in Table 1. The suspensions
larval sub-phases L3 and L4, recognized on the basis
containing 75 mg/ml UCG, out of the nine tests, three,
of size, were submitted to treatments with UCG.
aged between zero (used immediately after
The characteristics basically analyzed in the
preparation) and two days, produced 100% LM after
present study were the concentration and the age of the
one or two days’ treatment, and six tests with media
aqueous suspensions of UCG in number of days after
aged between three and 12 days produced from 70% to
their preparation. The treatments were carried out in
90% LM, after five to 13 days. These six tests yielded
two conditions: allowing the larvae to enter in contact
a total of 12 adults.
with the UCG deposits in the bottom of the
An increase in the concentration of the suspension
experimental glass or preventing them from doing so.
The effect of the addition or not of macerated fish food
also increased the percentage of tests with 100% LM,
to the experiments, for feeding larvae, was also
and its 100% mortality efficacy lasted longer,
analyzed.
extending to the age of nine days in the medium 300
The UCG suspensions involved the use of a
mg/ml UCG. For older suspensions of this
mixture of different brands of coffee.
concentration, the treatment time in which 100%
First Experiment. The larvae were placed directly into
mortality was obtained was extended to nine days,
the glasses bearing 200 ml of the suspension. In this
while for younger suspensions (up to four days old) the
case they were free to make contact both with the UCG
treatment time for 100% LM was 24h.
deposits in the glass bottom and with the supernatant
For more details, Table 2 shows the total results
liquid. The concentrations of dry UCG used were 75,
obtained in the First Experiment, in which the larvae



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2




BioAssay 2:5 (2007)

were free in the medium, being the normal way to use
days. The remaining three tests (aged two, seven and
the UCG.
nine days) showed from 70% to 80% mortality and
Statistical analysis using data from this Table
produced seven adults. In the two tests with the
showed that the linear correlation coefficients of UCG
addition of food (aged one and four days), LM was
suspension age and percentage larval mortality in 48h,
70% and 90% and the number of adults produced was
for every concentration used, was inverse: for 75mg/ml
four.
concentration, R= -0.758 (Figure 1A); for 150 mg/ml,
As in the First Experiment, in those tests carried
R= -0.854 (Figure 1B); for 225 mg/ml, R= -0.759
out without the addition of food, as the medium
(Figure 1C); and for 300mg/ml, R= -0.876 (Figure 1D).
concentration increased, the percentage of tests with
Second Experiment. Table 1 also shows that, in the
100% LM and the limit of the suspension age able to
experiments, in which the larvae were maintained in a
produce that rate also increased. The same was
sieve placed within the liquid part of the suspension,
observed in the media with the addition of food. The
two of the six tests using the medium without the
two tests using the medium with the highest UCG
addition of food, with 75 mg/ml UCG, aged zero and
concentration (300 mg/ml) and the addition of food
three days, produced 100% LM after six and four days’
produced 100% mortality. In the Second Experiment
treatment, respectively. A suspension with the same
the effect of the suspension on the 100% mortality, in
concentration, aged 15 days produced 100% LM in 19
tests without addition of food, lasted longer.


Figure 1. Scatter plots showing relation of used coffee ground suspension age and percentage larval mortality in 48
h treatment. Suspension concentrations: A. 75mg/ml, B. 150mg/ml, C. 225mg/ml and D. 300mg/ml.

In both experiments, we observed a variation in
(Laranja et al. 2003). In the same study UCG also
the treatment time that was necessary to produce 100%
affected developmental time increasing it.
mortality as the suspension age increased, but in the
In the present study, the effect of UCG on LM was
Second Experiment, in 225 and 300 mg/ml
analyzed in Experiments with two methods of treating
concentrations, as the suspension age increased, the
the larvae: (a) the larvae were set free in the medium so
treatment time required for 100% mortality decreased.
that they could enter into contact both with UCG grains
Previous data on treatments of A. aegypti eggs
deposited at the bottom of the recipient and with the
with UCG in the concentration 50 or 100 mg/ml
UCG suspension (the supernatant liquid) or (b) they
showed that this substance blocks the development in
were maintained in a fine-mesh sieve preventing them
the larval stage. Even when larvae do not die in three
from entering into contact with the deposit, but
or four days, the development is arrested in this stage
immersed in the liquid. The objective of these different



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3





S

E
o
f
c
f
Table 1 - Effect on Aedes aegypti larval mortality of used coffee ground (UCG) suspensions with different concentrations and ages after preparation, in two
e
i
e

c
t
d
experiments, the First with larvae free in the medium and the Second, with larvae inside a sieve immerse in the medium supernatant. Food was not added in the tests

o
a
f
d

c
e
of the experiment 1 (-), but was added to some tests of experiment 2 (+).
o

E

f
f
e
n
e
t
o

Larval Mortality

g
m
r
o
o
total
u
l
ó

100%
< 100%
n
UCG
d
g
Exp.
Food
N
number of
s
i
c

(mg/ml)
range
number of

o
a
tests
number of
suspension age
number of
suspension
mortality
range treat.
n

d

treat. time
adults

A
o
tests (%)
(days)
tests (%)
age (days)
range (%)
time (days)
e

B

(days)
produced
d
e
r
a

s
s

75
130
9
3 (33)
0 – 2
1 – 2

6 (67)
3 – 7; 12
70 – 90
5 – 13
12
a
i
l

e

g
y
p
150
130
9
4 (44)
0 – 3
1 – 2

5 (56)
4 – 7; 9
60 – 80
9 – 14
13
t
i

1
-
225
140
10
6 (60)
0 – 5
1

4 (40)
6; 7; 9; 12
60 – 90
7 – 11
10
w

300
140
10
9 (90)
0 – 7; 9
1 – 9

1 (10)
12
60
8
4
w
w
.
s
e
Control

water
50
1
0
-
-

1
0
70
30
15
b
.
o
r
g


.
b
75
60
6
3 (50)
0; 3; 15
4 – 19

3 (50)
2; 7; 9
70 – 80
14 – 34
7
r
/
b
i
o
a
150
70
7
3 (43)
0; 3; 15
2 – 14

4 (57)
2; 6; 8; 9
60 – 90
9 – 13
8
s
s
a
-
y
0; 2; 3; 6; 11;


225
80
8
7 (88)
2 – 9

1 (12)
13
80
16
2
12; 15
0; 2; 3; 6; 8; 9;
300
100
10
9 (90)
1 – 11

1 (10)
12
80
8
2
11; 13; 15
2
75
20
2
0 (0)
-
-
2 (100)
1; 4
70; 90
8 – 12
4
150
20
2
1 (50)
1
4

1 (50)
4
90
9
1
+
225
20
2
1 (50)
1
10

1 (50)
4
60
11
4
300
20
2
2 (100)
1; 4
2

0 (0)
-
-
-
-
G
u
i
-
water
30
1
0
-
-

1
0
27
37
22
r
a
d
Control
o

e
+
water
20
1
0
-
-

1
0
0
9
20

B

i
c
u
d
4
Exp. = experiment; treat. = treatment; N = number of larvae in each test.
o





BioAssay 2:5 (2007)


Table 2 -
Mortality (%) of Aedes aegypti in used coffee ground (UCG) suspensions with different concentrations
and ages after preparation, in the First Experiment (larvae free in the medium). Food was not added. MT =
Maximum Time (days) of mosquito surviving in the breeding sites; P = Mosquito death in the pupa stage. Each
test started with ten larvae, except the control (50 larvae).
Mortality percentage in the treatment time (days)
UCG
Suspension
MT Number
Test
(mg/ml)
age (days)
(days) of adults
1
2
3
4
5
6
7
8
9
10
1
0
98
20
-
-
-
-
-
-
-
-
-
-
2
1
80
100
-
-
-
-
-
-
-
-
-
-
3
2
100
-
-
-
-
-
-
-
-
-
-
-
4
3
50
60
60
60
70
70
70
70
70
70
10
3
75
5
4
50
50
50
60 (1P)
90
-
-
-
-
-
-
1
6
5
20
20
20
20
50
60
60
60
70
80
13
2
7
6
10
10
10
10
60
80
80
80
80
90
11
1
8
7
0
0
0
0
0
0
0
50
80
80
11
2
9
12
30
30
50
50
50
50
70
70
-
-
-
3
1
0
100
-
-
-
-
-
-
-
-
-
-
-
2
1
100
-
-
-
-
-
-
-
-
-
-
-
3
2
100
-
-
-
-
-
-
-
-
-
-
-
4
3
80
100
-
-
-
-
-
-
-
-
-
-
150
5
4
50
50
60
60
60
60
60
60
60
70
14
3
6
5
20
30
40
40
40
50
50
50
70
80
13
2
7
6
40
40
40
40
50
50
80
80
80
80
13
2
8
7
20
30
70
80
80
80
80
80
80
80
10
2
9
9
20
40
50
50
50
50
60
60
60
-
-
4
1
0
100
-
-
-
-
-
-
-
-
-
-
-
2
1
100
-
-
-
-
-
-
-
-
-
-
-
3
2
100
-
-
-
-
-
-
-
-
-
-
-
4
3
100
-
-
-
-
-
-
-
-
-
-
-
5
4
100
-
-
-
-
-
-
-
-
-
-
-
225
6
5
100
-
-
-
-
-
-
-
-
-
-
-
7
6
30
40
70
70
70
70
80
80
80
80
11
2
8
7
10
20
50
80
80
80
80
90
-
-
-
1
9
9
20
60
60
60
60
60
70
70
70
-
-
3
10
12
40
40
50
50
50
60
60
-
-
-
-
4
1
0
100
-
-
-
-
-
-
-
-
-
-
-
2
1
100
-
-
-
-
-
-
-
-
-
-
-
3
2
100
-
-
-
-
-
-
-
-
-
-
-
4
3
100
-
-
-
-
-
-
-
-
-
-
-
5
4
100
-
-
-
-
-
-
-
-
-
-
-
300
6
5
90
90
100
-
-
-
-
-
-
-
-
-
7
6
60
70
70
80 (1P) 100
-
-
-
-
-
-
-
8
7
40
60
60
70
70
80 (1P)
90
90
100
-
-
-
9
9
80
80
80
80
80
90
100
-
-
-
-
-
10
12
20
20
20
20
20
30
50 (2P) 60 (1P)
-
-
-
4
Control
1
0
8
14
20
34
34
34
34
36
36
36
30
15




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5




Effect of coffee grounds on Aedes aegypti
Guirado e Bicudo

treatments was to test the supposed “mechanical
The statistical analysis of data obtained in the First
effect” of UCG suggested by FUNASA (FUNDAÇÃO
experiment, considering the percentage of larval
NACIONAL DE SAÚDE do Ministério da Saúde) in
mortality in the first 48h treatment, in every UCG
the Technical Report published on the internet, on
concentration, and the suspension age showed inverse
21/11/2001 (no more available), concerning the use of
linear correlation coefficients, reinforcing the general
UCG, that had been propagated in the media.
observation that as UCG suspension age increased, the
The present results shows that the effect of UCG
efficiency decreased, but more slowly in the greater
on producing 100% LM is independent of contact
concentrations.
between larvae and the deposit at the bottom of the
The UCG medium at 300 mg/ml concentration
container. The mortality actually seems to be due to
was the most efficient for treatment of larvae. In both
toxic compounds of UCG present in both the
experiments, adults began to be produced in this
supernatant liquid and the deposit. UCG includes, in
medium only when the suspension was 12 days old.
addition to caffeine, anti-physiological components
Besides, the number of adults produced in both cases,
such as tannins, chlorogenic acid, caffeic acid and
after this time, in tests without addition of food was
potassium in excess (Brenes 1979). Laranja et al.
smaller than that produced in the media with the lower
(2003) considered that some of these components of
UCG concentrations (75 and 150 mg/ml) during the
UCG, in addition to caffeine, might be involved in the
entire time of treatment.
detrimental effects on A. aegypti because the effects of
Normally, in natural breeding sites, larvae feed on
UCG on the esterase pattern differed from those of
aquatic microorganisms (bacteria, algae and yeasts),
caffeine when used isolated. In fact, data in literature
pollen, molts, carcasses of dead larvae and a wide
shows that tannins have larvicidal activity (Silva et al.
variety of organic detritus that exists in the medium
2004). The property of the tannins to interact with
(Jones 1978). In the laboratory, because tap water is
proteins makes them highly toxic (Simões et al. 2001,
used, bacteria and carcasses of dead larvae are the main
in Silva et al. 2004).
food source when fish food is not added. In the present
A difference between the First and the Second
study, the addition of food to UCG media did not alter
Experiments was observed as to the duration of the
the results substantially. In UCG at 300 mg/ml (aged
effect of UCG on LM. The efficiency of the lowest
one and four days), no adult was produced when fish
concentrations of UCG (75 and 150 mg/ml), evaluated
food was added. In the lower concentrations, there was,
in terms of the treatment time necessary to cause 100%
similarly, no increase in the number of adults produced,
LM was smaller in the Second Experiment (1 to 2 days,
when compared with the tests without the addition of
in the First Experiment; 4 to 19 in the Second
food.
Experiment). These observations suggest that, in low
It is interesting to note that, in the Second
UCG concentrations, since the amount of toxic
Experiment (larvae in the sieve), adults were not
components is probably greater in the deposit than in
produced in the three smaller concentrations (75, 150
and 225 mg/ml) of 15-day-old suspensions and in the
the supernatant, direct contact with the deposit
300 mg/ml concentration aged 13 days. This result may
increases contact with the toxic agents. In addition, the
be attributed to water evaporation in the experimental
larvae feed on the UCG grains (as shown by the
breeding sites, which were only covered with a fine-
observation of black material inside their digestive
mesh fabric. The evaporation reduced the amount of
tract), and this certainly increases the effect. The
water, which may have impaired the survival of the
movement up and down of the larvae in the medium
larvae inside the sieve.
also may help to distribute the toxic components in the
In the controls without addition of food, the larva
experimental vial when they are free.
– adult developmental time increased (30 and 37 days),
Also in the lower concentrations (75 and 150
in comparison to the control with addition of food (9
mg/ml), in general, as the suspension age increased, the
days). The low productivity of adults in the control of
treatment time necessary for the larvae to die also
the First Experiment may be due to the fact that L3
increased (reaching 14 days in the First Experiment
larvae (more sensitive to the small availability of food)
and 34 days in the Second Experiment). Apparently,
were predominantly used in that test.
UCG components, which are toxic for A. aegypti
The present results reinforce those obtained by
become degraded and lose their effect as time goes by.
Laranja et al. (2003) showing that UCG causes larval
In the two experiments, tests using the higher
mortality in A. aegypti and then may be considered a
UCG concentrations (225 and 300 mg/ml) and no
potential agent for alternative control, when used in the
addition of food showed an increased percentage with
appropriate concentration and with the addition of new
100% LM (60% and 90% in the First Experiment and
UCG suspension at appropriate intervals. The present
88% and 90% in the Second, respectively) and showed
study shows that 300 mg/ml is a secure concentration.
an increased duration of the UCG effect (reaching nine
In relation to the interval, although the present results
days in the First Experiment and 11 days in the Second
have shown that, at this concentration, 100% mortality
Experiment for 300 mg/ml).
occurs in media up to 9 days old, we consider it safer



Sociedade Entomológica do Brasil
www.seb.org.br/bioassay
6




BioAssay 2:5 (2007)

to recommend the use of seven-day intervals for the
Jones JC. Feeding behavior of mosquito. Sci Am.
addition of new UCG suspension.
1978; 238: 112-20.
Thus, although the elimination of the breeding
Laranja A.T., A.J. Manzatto & H.E.M.C. Bicudo. 2003.
sites remains being the best way to control A. aegypti
Effects of caffeine and used coffee grounds on
population size, the present data reinforces the validity
biological features of Aedes aegypti (Diptera,
of considering UCG preparations as possible auxiliary
Culicidae) and their possible use in alternative
in the alternative control of this mosquito. UCG might
control. Genet. Mol. Biol. 26: 419-429.
be recommended mainly to be used in gardens inside
Macoris M.L.G., M.T.M. Andrighetti, L. Takaku C.M.
Bromeliaceae, in the dishes under vases (when the
Glasser, V. Garbeloto & J.E. Bracco. 2003.
dishes cannot be discarded) and over the land in the
Resistance of Aedes aegypti from the state of São
vases. UCG has the advantage of being free of cost (it
Paulo, Brazil, to organophosphates insecticides.
is normally through out after the drink preparation) and
Mem. Inst. Oswaldo Cruz. 98: 703-708.
Marzochi, K.B.F. 1994. Dengue in Brazil – situation,
used by many people as fertilizer for plants.
transmission and control: a proposal for ecological

control. Mem. Inst. Oswaldo Cruz. 89: 235-245.
Acknowledgements
Promsiri, S., A. Naksathit, M. Kruatrachue & U.
Thanks are due to SUCEN (Superintendência de
Thavara. 2006. Evaluations of larvicidal activity of
Controle de Endemias) of São José do Rio Preto - SP,
medicinal plant extracts to Aedes aegypti (Diptera:
for providing mosquitoes, to Dr. Antônio José
Culicidae) and other effects on a non target fish. J.
Manzatto for help with the statistical analysis and to
Insect Sci. 13: 179.
the Zoology and Botany Department of IBILCE for
Silva, H.H.G., I.G. Silva, R.M.G. Santos, E. Rodrigues
providing the mice for feeding the mosquitoes.
Fillho & C.N. Elias. 2004. Atividade larvicida de

taninos isolados de Magonia pubescens St. Hil.
(Sapindaceae) sobre
References
Aedes aegypti (Diptera,
Culicidae). Rev. Soc. Bras. Med. Trop. 37: 1-8.
Brenes, R.A.G. 1979. Processing of coffee pulp:
Slosek, J. 1986. Aedes aegypti mosquitoes in the
chemical treatments. In: Coffee pulp –
Americas: a review of their interactions with the
Composition, technology, and utilization. Institute
human population. Soc. Sci. Med. 23: 249-257.
of Nutrition of Central America and Panama. p. 72-
Sousa-Polezzi R.C. & H.E.M.C. Bicudo. 2004. Effect
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of phenobarbital on inducing insecticide tolerance
Chauhan LK, N. Pant, S.K. Gupta & S.P. Srivastava.
and esterase changes in Aedes aegypti (Diptera:
2000. Induction of chromosome aberrations,
Culicidae). Genet. Mol. Biol. 27: 275-283.
micronucleus formation and sperm abnormalities in
Tian Y, Ishikawa H & Yamauchi T 2000. Analysis of
mouse following carbofuran exposure. Mutat. Res.
cytogenetic and developmental effects on pre-
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implantation, mid-gestation and near-term mouse
Furtado, R.F., M.G.A. Lima, M. Andrade Neto, J.N.S.
embryos after treatment with trichlorfon during
Bezerra & M.G.S. Silva. 2005. Larvicidal activity
zygote stage. Mutat. Res. 471: 37-44.
of essential oils against Aedes aegypti L. (Diptera:
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Culicidae). Neotrop. Entomol. 34: 843-847.
New Jersey, USA.




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www.seb.org.br/bioassay
7


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