Neotrop Entomol (2016) 45:604–611

DOI 10.1007/s13744-016-0409-7

PEST MANAGEMENT

Toxicity of Piper aduncum (Piperaceae) Essential Oil

Against Euschistus heros (F.) (Hemiptera: Pentatomidae)
and Non-Effect on Egg Parasitoids

LM TURCHEN1,4, LP PITON2, EL DALL’OGLIO3, AR BUTNARIU2, MJB PEREIRA2

1
Dept of Entomology, Univ Federal de Viçosa, Viçosa, MG, Brasil
2
Dept of Agronomy, Univ do Estado de Mato Grosso, Tangará da Serra, MT, Brasil
3
Dept of Chemistry, Univ Federal do Mato Grosso, Cuiabá, MT, Brasil

Keywords
Biological control, brown stink bug,
phytoinsecticide, selectivity, sustainable
pest control
Correspondence
LM Turchen, Depto de Entomologia,
Programa de Pós Graduação em
Entomologia, Univ Federal de Viçosa,
Viçosa, MG 36570-000, Brasil; leonardo.
turchen@ufv.br
Edited by Moisés J Zotti – UFPel
Received 3 October 2015 and accepted 15
April 2016
Published online: 2 June 2016
* Sociedade Entomológica do Brasil 2016
Abstract
Plant essential oils have been recognized as significant natural resources for
insecticides. Herein, we have assessed the toxicity of the essential oil of Piper
aduncum (Piperaceae) against Euschistus heros (F.) (Hemiptera:
Pentatomidae), a key soybean pest in Neotropical America. In addition, we
have assessed its effect on the performance of egg parasitoids. The essential
oil was obtained from the leaves of P. aduncum via hydrodistillation.
Subsequently, bioassays of the concentration response to eggs (contact and
immersion methods), nymphs, and adults (topical application) were conduct-
ed, to assess the lethal effects on the stink bug. We also evaluated the
performance of parasitism and adult emergence of egg parasitoids, when
the host eggs were treated with essential oil. In the egg bioassay, both expo-
sure methods were efficient for unviable eggs (immersion
LC50 = 15.64 mg mL−1; contact LC50 = 21.29 mg mL−1), with the highlight on
the immersion method. The bioassay with nymphs indicated a higher toxicity
of essential oil, with lower concentrations (LC50 = 11.37 mg mL−1) being re-
quired to cause the death of insects. For adults, a reduction in
survival of insects was observed, and consequently, there was a
reduction in the number of individuals in the next generation.
Although the essential oil was toxic to E. heros, it exhibited lower
toxicity for egg parasitoids, as there was no effect on parasitism and
the emergence of wasps. We discuss likely explanations for such
selectivity. In summary, we found that the essential oil was promis-
ing for the control of E. heros, because it caused deleterious effects
at all development stages of the stink bug and had no effect on
parasitism and emergence of the egg parasitoids, which suggested
compatibility with biological control.

Introduction parasitoids (Turchen et al 2016). The integration of other

Insecticides are generally used for control of arthropod pests,
mainly because of the efficiency of these products. However,
these are often used incorrectly, which can contribute to
selection of resistant populations of insects (Sosa-Gomez &
Silva 2010) and suppression of natural enemies, such as egg
control strategies, such as phytoinsecticides and biological
controls, can be an alternative for reducing pesticide use
(Biondi et al 2013, Turchen et al 2014). In this context, plants
with insecticidal properties have been investigated as a strat-
egy for pest control, with particular emphasis on substances
of secondary metabolism, which are known to act as
Toxicity of Piper Essential Oil on Stink Bug and Egg Parasitoids
repellents or deterrents (oviposition or feeding) or may even
be lethal to herbivorous insects (Mithofer & Boland 2012).
Furthermore, they have different modes of action (El-Wakeil
2013), which make them interesting for researchers who aim
to develop new insecticidal molecules.
The Piperaceae family has been investigated for control-
ling insect pests, as its insecticide action has proved to be
efficient for many insect orders. For instance, Piper aduncum
(Piperaceae) has an insecticide effect against Coleptera
(Estrela et al 2006, Fazolin et al 2007), Hymenoptera
(Souto et al 2012), Diptera (Bernard et al 1995, Misni et al
2011), and Hemiptera (Piton et al 2014, Volpe et al 2015). It is
known that P. aduncum is commonly used to obtain an es-
sential oil with active compounds, such as terpenoids and
phenylpropanoids, which can act as repellents and insecti-
cides (Bernard et al 1995, Assis et al 2013, Souto et al 2012).
However, little is known about the effect of the essential oil
against the stink bug and its natural enemies. This study is
significant as it provides information about the possible com-
patibility between phytoinsecticides and biological control
methods.
In this context, we used as a model the Neotropical brown
stink bug, Euschistus heros (F.) (Hemiptera: Pentatomidae),
which is a pest of economic importance in soybean crops
(Corrêa-Ferreira & Azevedo 2002, Silva et al 2012), and its
natural enemies, Telenomus podisi (Ashmead)
(Hymenoptera: Platygastridae) and Trissolcus urichi
(Crawford) (Hymenoptera: Platygastridae) (Golin et al 2011,
Laumann et al 2010, Turchen et al 2014, 2016), to assess (1)
the toxicity of the essential oil against eggs, nymphs, and
adults of Pentatomidae and (2) its effect on parasitism and
the emergence of egg parasitoids.
Material and Methods
Insect populations
The populations of the egg parasitoids (T. podisi and T. urichi)
and of the brown stink bug, E. heros, were collected in soy-
bean fields in the municipality of Tangará da Serra (State of
Mato Grosso, Brazil) and reared in controlled environmental
conditions at 26 ± 2°C temperature, 70 ± 10% relative humid-
ity, and L12/D12 photophase, the same conditions in which
the bioassays were performed. The egg parasitoids were
identified by Dr. Norman Johnson and deposited in the C.A.
Triplehorn Insect Collection at the Ohio State University
(Columbus, OH, US), as voucher specimens (T. pod
isi = OSUC 480194, T. urichi = OSUC 480207). The wasps were
maintained in 15-mL glass test tubes sealed with cotton balls
containing thin honey strips for adult feeding, as described
by Peres & Corrêa-Ferreira (2004). The wasps multiplied in
the newly laid eggs of the brown stink bug (<24 h). The
605
brown stink bugs were mass reared as described elsewhere
(Silva et al 2008, 2011).
Plant material and extraction of volatile components
Plants of P. aduncum were collected from Tangará da Serra,
MT (14°29ʹS and 57°54ʹW), in December 2011 and January
2012. These plants were identified and deposited in the
Tangará herbarium (TANG) with a voucher (registration no.
1738). For the preparation of the extract, the leaves of
P. aduncum were dried in a drying oven with forced air cir-
culation for 96 h at 37°C, following the methodology of Prista
et al (1981). After drying, the leaves were ground to a vege-
table powder using a knife mill and a sieve with a 1-mm
diameter mesh. The powder obtained was subjected to
hydrodistillation (100 g by 3 h), with an extractor type
Clevenger. The hydrolat was partitioned with a separatory
funnel using dichloromethane (DCM), and the obtained or-
ganic fractions were combined and dried over anhydrous
calcium chloride (CaCl2). The salt was removed by vacuum
filtration and the solvent evaporated at room temperature.
Chemical composition of the volatile material
The oil was analyzed by gas chromatography and mass spec-
trometry (GC-MS) (Shimadzu, QP 5050A), with the following
conditions: column, DB-5 fused silica (30 m × 0.25 mm i.d.,
0.25 μm film thickness); carrier gas helium, adjusted to a
linear velocity of 1.2 mL/min, temperature programmed at
110–140°C (5°C/min), 140–290°C (20°C/min), and 290–330
(5°C/min), and mass spectra, 70 eV (in El mode). The individ-
ual components were identified by comparison of both the
mass spectra and their GC retention data with those authen-
tic compounds previously analyzed and were stored in the
data system. Other identifications were made by comparing
the mass spectra with those in the Wiley data system librar-
ies (software class 5 k) and were cited in the literature. The
retention time (RT) is exhibited in Table 1.
Stink bug bioassay
Egg, nymph, and adult bioassays were conducted in a
completely randomized design at concentrations of 5, 10,
20, 40, and 80 mg mL−1 of the essential oil of P. aduncum
and Acetone P.A. (control), with 10 replications/treatment.
For bioassays with eggs, 30 eggs (<24-h old) were used per
repetition (n = 300 eggs/treatment). For the bioassay with
nymphs, five third-instar individuals were used per repetition
(n = 50 nymphs/treatment), and for the bioassay with adults,
one pair of E. heros (<24-h old) was used per repetition
(n = 20 adults/treatment).
All bioassays were conducted following the methodology
described by Piton et al (2014). The eggs were treated using
606 Turchen et al

Table 1 Constituents identified, retention time (RT), and number of unviable eggs (10 days), dead nymphs (15 days),
concentration of compound (%) in the essential oil of Piper aduncum
and the ones that survive as well as the fecundity (number of
collected in Tangará da Serra-MT.
eggs/female) and fertility (number of eggs viable/female) of
Constituentsa RTa Percentage adults until death were assessed.
α-Pinene 1.938 0.23
Egg parasitoid bioassay
β-Myrcene 2.408 0.33
γ-Terpinene 3.319 1.57
Another set of bioassays was conducted following a
o-Cymene 3.504 4.20
completely randomized design, with the concentrations at
Cyclohexene 3.567 0.87
5, 10, 20, and 40 mg mL−1 of essential oil of P. aduncum
6-Methyl-5-hepten-2-one 3.913 0.60 and acetone P.A. (control) and five replications/treatment
3-Cyclohexene-1-carboxaldehyde 4.808 0.31 (n = 50 eggs/treatment). To assess the parasitism of
α-Cubebene 5.074 0.64 T. podisi and T. urichi on eggs treated with essential oil, an
Hexadecane 5.289 3.61 egg mass (>24-h old) of 10 eggs each was immersed in the
Tridecane 5.297 6.26 treatment for 10 s. After immersion, the eggs were placed on
Aromadendrene 5.997 9.20 a glass surface to dry for 1 h. Next, the eggs were individually
(Z,E)-α-Farnesene 6.005 2.40 offered to female parasitoids (adults), according to the meth-
2-Undecanethiol 2-methyl 6.334 0.19 odology of Turchen et al (2014). After 24 h of parasitism, the
α-Gurjunene 6.641 1.00 females were removed and the eggs maintained until emer-
(Z,Z)-α-Farnesene 6.764 0.70 gence of parasitoids. The eggs from which wasps did not
α-Serinene 7.087 7.31 emerge were dissected under a stereomicroscope for confir-
γ-Elemene 7.158 4.58 mation of parasitism. The number of parasitized eggs and
β-Gurjunene 7.438 1.08 wasps that emerged was recorded.
p-Cymen-8-ol 8.210 0.66
1-Nonyne 8.956 0.43 Statistical analysis
Dipentene dioxide 9.398 0.83
Caryophyllene oxide 9.514 2.84 Normality and homoscedasticity of the data obtained were
Humulene epoxide II 10.008 0.48 checked using the Shapiro-Wilk and Bartlett tests, respective-
Nerolidol 10.138 1.94 ly. The number of unviable eggs (contact and immersion),
1-Methyl-1,2-cyclohexene oxide 10.279 0.36 mortality (nymph), number of eggs (fecundity), and number
3-Pinanol 10.362 1.23 of nymphs hatched (fertility) were subjected to analyses of
2,5-Dimethyl-3-hexyne-2,5-diol 10.443 0.57 deviance and general linear model (GLM) with Poisson distri-
Ledol 10.487 0.31 bution and log as the function link. Significant treatments
Spathulenol 10.722 3.00 were compared by contrasts (p < 0.05). Lethal concentra-
Limonene 10.914 0.89 tions (LC50) were estimated for eggs and nymphs with
Viridiflorol 11.045 0.84 Probit analysis. The survival of adults (males and females,
T-muurolol 11.166 0.36 separately) over time was subjected to survival analyses,
Myristicin 11.215 30.03 using the Kaplan-Meier (Gehan-Breslow) estimators, and
α-Bisabolol 11.246 0.23 the survival curves were compared using the Holm-Sidak
β-Eudesmol 11.374 1.28 method (p < 0.05), always using the R software version,
Dillapiole 12.164 8.43 3.1.1 (R Development Core Team 2014).
α-Limonene diepoxide 13.264 0.21
Monoterpenes – 09.90
Sesquiterpenes – 48.06 Results
Phenylpropanoids – 38.46
Total 96.42
Essential oil composition

Text in bold is for highlighting the major compounds.
a
Obtained from the GC library.
two methods: (1) immersion and (2) contact with an impreg-
nated surface. The nymphs and adults were treated by top-
ical application of l and 5μL treatments, respectively. The
The average yield of essential oil was 10.2 g/kg. The chro-
matograph analysis of P. aduncum essential oil showed a
standard chemical of compounds, as expected, for this spe-
cies (Table 1); however, it was recognized that the chemical
concentration of compounds was variable in relation to other
researches. Here, it was observed that the more abundant
Toxicity of Piper Essential Oil on Stink Bug and Egg Parasitoids 607

compounds were myristicin (30.03%), aromadendrene
(9.20%), dillapiole (8.43%), α-serine (7.31%), tridecane
(6.26%), γ-elemene (4.58), and o-cymene (4.20%). These
compounds are highlighted in bold in Table 1.
Stink bug bioassay
The application of the essential oil of P. aduncum on eggs,
nymphs, and adults of E. heros exhibited promising results
for the control of this insect as it caused deleterious effects at
all development stages of the stink bug. For the egg bioassay,
a significant difference was observed between contamina-
tion methods (χ2 = 6.15; df = 1, 113; p < 0.05), the higher being
the mean of unviable eggs in the immersion method (17.63
± 8.8a), when compared to the contact method (15.78
± 7.7b).
The concentration mortality bioassay showed that the
number of unviable eggs differed between treatments used
in the immersion method (χ2 = 311.75, df = 5. 54, p < 0.01) and
contact method (χ2 = 265.41; df = 5, 54; p < 0.01). The per-
centage of unviable eggs was between 54 and 96% in the
immersion method and between 47.7 and 70.7% in the con-
tact method, in concentrations of 5.0 to 80.0 mg mL−1, re-
spectively. Although a lower proportion of unviable eggs was
observed for the contact method, a significant increase in
unviable eggs was noticed, with increasing concentrations,
regardless of the method of application (immersion or con-
tact) (Table 2). The lethal concentrations (IC 95%) estimated
for eggs treated with the essential oil of P. aduncum were
LC50 = 15.64 mg mL−1 (11.59–19.68) and LC50 = 21.29 mg mL−1
(20.50–22.08) for the immersion method and contact meth-
od, respectively. These results highlighted the immersion
method, because it needed a lower concentration, as com-
pared to the contact method.
In the bioassay with nymphs, the number of dead nymphs
differed between treatments (χ2 = 72.54; df = 5, 54; p < 0,01),
with mortality from 62 to 100%, in concentrations of 5.0 to
80.0 mg mL−1. A significant increase in the number of dead
nymphs was noted, with increasing concentrations, similar to
the results found for the bioassay with eggs (Table 2). The
lethal concentration (IC 95%) estimated for nymphs treated
with essential oil of P. aduncum was LC50 = 11.37 mg mL−1
(8.85–14.12) and LC90 = 38.95 mg mL−1 (29.15–60.83), which
indicated a higher toxicity of essential oil.
Adult survival, when treated with essential oil, exhibited
significant differences among treatments for males (Gehan–
Breslow = 66.61, df = 5, p < 0.001) and females (Gehan–
Breslow = 58.75, df = 5, p < 0.001). The longevity of males
treated with the essential oil of P. aduncum was concentra-
tion-dependent, with a survival of 1 to 86 days at concentra-
tions of 5 to 80 mg mL−1, which contrasted with that of the
control (126 days) (Fig 1a). Similar results were observed in
the longevity of females treated the essential oil, with a sur-
vival of 1 to 84 days at concentrations of 5 to 80 mg mL−1,
when compared with the control (96 days) (Fig 1b).
The median lethal time (LT50) of insects (males and fe-
males) was lower for insects treated with essential oil, when
compared with the control, with the highlight on concentra-
tions >20 mg mL−1, which exhibit LT50 <6 days for males and
females (Table 3). In addition, the surviving females exhibited
a significant reduction in fertility and fecundity when com-
pared to the control (Table 3). However, this was because of
the low survival rate of the females treated, which conse-
quently reduced the number of eggs.
Bioassay with egg parasitoids
When the host eggs were treated with essential oil and of-
fered to the egg parasitoid female (adults), no significant
difference was found for egg parasitism between treatments
(χ2 = 0.92; df = 4, 45; p = 0.92) and between the species stud-
ied (T. podisi or T. urichi) (χ2 = 2.42; df = 1, 44; p = 0.12).

Table 2 Mean of unviable eggs

treated by the method of Treatments Numbera Unviable eggs mortality
immersion and contact and
mortality of nymphs Euschistus (Immersion method) (Contact method)
heros treated with essential oil of
Piper aduncum. (X ± SD)b % (X ± SD)b % (X ± SD)b %

80.0 mg mL−1 10 28.8 ± 1.47a 96.0 21.2 ± 3.58a 70.7 5.00 ± 0.00a 100.0
40.0 mg mL−1 10 22.1 ± 2.99b 73.7 21.0 ± 2.78a 70.0 5.00 ± 0.00a 100.0
20.0 mg mL−1 10 19.4 ± 3.43bc 64.7 20.2 ± 4.75ab 67.3 5.00 ± 0.00a 100.0
10.0 mg mL−1 10 17.6 ± 4.55c 58.7 16.6 ± 3.53bc 55.3 4.30 ± 1.41ab 86.0
5.0 mg mL−1 10 16.2 ± 4.58c 54.0 14.3 ± 3.46c 47.7 3.10 ± 0.99b 62.0
Acetone P.A. 10 1.70 ± 1.05d 5.7 1.40 ± 1.89d 4.7 0.20 ± 0.42c 4.0
a
Number of repetition for biassay (postures: 30 eggs per repetition; nymphs: 5 insects per repetition).
b
Mean ± standard deviations followed by the same letter in the column do not differ by multiple comparison for
contrast test (P = 0.05).
608 Turchen et al

Fig 1 Survival curves of

Euschistus heros males (a) and
females (b), when treated with
essential oil of Piper aduncum.

Similarly, no difference was observed in the number of par-
asitoids that had emerged, both for the oil concentrations
(χ2 = 7.96; df = 4, 45; p = 0.09) and for the species studied
(χ2 = 0.30; df = 1, 44; p = 0.58) (Fig 2).
Discussion
The compounds that were present in the essential oil of
P. aduncum were previously reported in other researches
with chromatographic studies (e.g., Maia et al 1998, Estrela
et al 2006, Almeida et al 2009, Fazolin et al 2005, 2007,
Souto et al 2012, Potzernheim et al 2012, Volpe et al 2015).
However, as expected, we did not find a quantitative stan-
dard of compounds between studies; for example, in this
research, myristicin (30.03%) was the major compound in
the essential oil and not dillapiole (8.43%), as expected.
The explanation for these results may be on account of in-
numerous factors (e.g., region, seasonal, nutritional, and
others), which can interfere with the proportion of com-
pounds in a plant (Almeida et al 2009, Assis et al 2013).
The essential oil of P. aduncum exhibited promising results
for control of the brown stink bug, as it caused deleterious
effects at all developmental stages of E. heros. These essen-
tial oils and their constituents are potentially active as insec-
ticides, because they have terpenoid and phenylpropanoid
compounds (see Table 1), which are responsible for most of
the repellent and insecticidal effects (Scott et al 2007, Souto
et al 2012). Research that shows that the lethal effect of
phytoinsecticides on stink bugs (Hemiptera: Pentatomidae)
is scarce, however, has been reported for Azadirachta indica
(Meliaceae) against Oebalus poecilus (Dallas) (Pinheiro &

Table 3 Survival of male and

female (days), fecundity (number Treatments Numbera Survivalb Fecundityc Fertilityc
of eggs), and fertility (number of
nymphs hatched) of adults Male Female
Euschistus heros treated with
essential oil of Piper aduncum. 80.0 mg mL−1 10 01.00 ± 00.00c 01.00 ± 00.00c – –
40.0 mg mL−1 10 01.50 ± 00.26c 01.20 ± 00.20c – –
20.0 mg mL−1 10 05.00 ± 02.58c 04.70 ± 03.70c 08.60 ± 27.19c 00.20 ± 00.63c
10.0 mg mL−1 10 35.10 ± 08.76b 08.00 ± 03.56b 07.30 ± 20.12c 06.40 ± 19.20c
05.0 mg mL−1 10 43.30 ± 10.82b 43.00 ± 09.29a 163.20 ± 175.73b 114.20 ± 125.20b
Acetone P.A. 10 85.10 ± 10.21a 67.80 ± 08.06a 224.40 ± 145.16a 170.90 ± 120.05a
a
Number of repetition for biassay (one pair of E. heros/repetition n = 20 adults/treatment).
b
Median lethal time ± standard errors followed by the same letter in the column do not differ by multiple
comparison Holm-Sidak (p = 0.05).
c
Mean ± standard deviations followed by the same letter in the column do not differ by multiple comparison
contrast test (p = 0.05).
Toxicity of Piper Essential Oil on Stink Bug and Egg Parasitoids
Fig 2 Performance (parasitism
and emergence) of Telenomus
podisi and Trissolcus urichi on
host eggs treated with essential
oil of Piper aduncum. ns no
significant difference.
Quintela 2010), Nezara viridula (Linnaeus) (Abudulai
et al 2003, Singha et al 2007), and E. heros (Silva
et al 2013) and for Annonaceae against Tibraca
limbativentris Stal. (Krinski & Massaroli 2014),
Dichelops melacanthus Dallas (Souza et al 2007), and
E. heros (Silva et al 2013, Turchen et al 2014).
The ovicidal effect of P. aduncum on E. heros eggs was
previously reported by Piton et al (2014), using the crude
extract of this plant. Similar results were observed in this
research with essential oil, as there was higher percentage
of unviable eggs in the immersion method, when compared
to contamination by the contact method. This result was
expected, because, in the immersion method, the surface
contact was higher when compared to the contact method.
It was believed that essential oil increased the efficiency,
because of a greater affinity of the eggs to lipophilic com-
pounds, which allowed the entry of this compound into the
chorion layers. This explained the higher efficiency of the
immersion method, when compared to the contact method.
Another hypothesis that can be related to unviable eggs is
the fumigant action, as suggested by Coitinho et al (2011) and
Sahayaraj & Kalidas (2011), which is consistent with the re-
sults shown in the contact method.
The essential oil was highly toxic for nymphs of E. heros.
This could be clearly seen, as a low lethal concentration was
required to cause the death of the stink bug
(LC50 = 11.37 mg mL−1). Piton et al (2014) exhibited similar
results, although the amount of the crude extract required
to cause death in 50% of the nymphs was greater than
40 mg mL−1. These results indicated that by using the essen-
tial oil of P. aduncum there was more efficiency in pest
control.
For adults, a decrease in the survival rate (males and fe-
males), number of eggs, and nymphs hatched in the next
generation was observed when treated with essential oil.
These results were significant in pest management, as that
drastic reduction in survival rate (i.e., 80 and 40 mg mL−1)
could consequently reduce the damage caused by the stink
bug in soybean crops, in addition to suppressing the next
generations, because this insect needed 10 days (average)
609
for sexual maturity (Costa et al 1998, Piton et al 2014).
Although concentrations lower than 20 mg mL−1 do not
cause death of adults before they are 10 days old (pre-
oviposition period) (Fig 1), a significant reduction in the num-
ber of eggs (fecundity) and the number of nymphs hatched
(fertility) was observed (Table 3), which indicated the efficacy
of essential oil in compromising the population growth of
E. heros.
The reduction in longevity and inhibition of oviposition of
females of Hemiptera are also reported in N. viridula, when
treated with neem-based products containing compounds
rich in azadirachtin (95%) (Riba et al 2003) and in E. heros,
when treated with the P. aduncum extract (Piton et al 2014).
However, it is evident that essential oil shows promising re-
sults, in relation to other researches. The toxic effect of
P. aduncum against nymphs and adults of E. heros is likely
associated with the action of the bioactive compounds, such
as myristicin (essential oil = 30.03%, see Table 1), responsible
for the insecticidal and antifungal activity, as suggested by
Morais et al (2007) and Boulogne et al (2012). That said,
other compounds may be responsible, such as dillapiole (es-
sential oil = 8.43%, see Table 1) which is responsible for toxic
effects in other insects, as suggested by Estrela et al (2006),
Fazolin et al (2007), Fazolin et al (2005), and Piton et al
(2014). However, the possibility of synergistic action between
compounds cannot be discarded, as suggested by Nerio et al
(2010); therefore, further investigation is needed to confirm
this hypothesis.
The essential oil of P. aduncum was selective to T. podisi
and T. urichi, because it did not affect parasitism and the
emergence of wasps. It is believed that the non-
effectiveness of essential oil on the egg parasitoid can be
associated with rapid degradation of the phytoinsecticide,
but more research is needed to confirm this hypothesis.
Similar results were observed by Abudulai & Shepard
(2003) and Mitchell et al (2004), when testing a base insec-
ticide for azadirachtin on the parasitoid Trissolcus basalis
Wollaston (Hymenoptera: Scelionidae) in the eggs of
N. viridula and Gryon fulviventris (Crawford) (Hymenoptera:
Scelionidae) and in the eggs of Clavigralla scutellaris
610
(Westwood) (Hemiptera: Coreidae), with parasitism of 89
and 84.7%, respectively. However, contrasting results were
observed in the bioassay with azadirachtin and the Annona
extract, as there was a lower parasitism rate, when the egg
host was treated, indicating deterrence of oviposition
(Smaniotto et al 2013, Turchen et al 2014) and negative sub-
lethal effects on parasitism and population growth of the egg
parasitoid (Turchen et al 2016).
The emergence of egg parasitoids (T. podisi and T. urichi)
was not affected when the host eggs were treated with es-
sential oil. The intrinsic traits of the egg parasitoid, as it de-
velops inside the host egg (12 to 13 days—pre-imaginal peri-
od), protect the immature stages from external conditions
(Orr 1988, Austin et al 2005), although it may allow its asso-
ciation with insecticides or phytoinsecticides, favoring eco-
logical selectivity (Foerster 2002, Turchen et al 2014).
However, the residual period of phytoinsecticides should be
less than the pre-imaginal period, in order not to impair the
maintenance of wasps and, consequently, the ecological ser-
vices they provide, as seen in this research.
In summary, these results demonstrate the potential
phytoinsecticide properties of the essential oil of
P. aduncum against the brown stink bug (E. heros) and its
non-effect on the parasitism and emergence of T. podisi and
T. urichi. Thus, we can suggest that the essential oil of
P. aduncum is compatible with the biological control.
However, further research in semi-fields and fields are need-
ed, for decision making in management of this pest.
Acknowledgments The authors thanks the Fundação de Amparo a
Pesquisa do Estado de Mato Grosso (FAPEMAT) for financial assistance
(process number: 163079/2014), the Conselho Nacional de
Desenvolvimento Científico e Tecnológico (CNPq) for providing
Masters scholarship (process number: 134392/2015-9), and Dr.
Norman Johnson for confirmation of species of Telenomus podisi and
Trissolcus urichi.
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