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Short Communication

Rice weevil response to basil oil fumigation


Peter A. Follett , Keao Rivera-Leong, Roxana Myers
U.S. Pacic Basin Agricultural Research Center, USDA-ARS, 64 Nowelo St., Hilo, HI 96720, USA
a b s t r a c t a r t i c l e i n f o
Article history:
Received 29 July 2013
Accepted 29 November 2013
Keywords:
Sitophilus oryzae
Plant essential oil
Quarantine treatment
Ocimum basilicum
Rice
Pest management
Basil oil, Ocimum basilicum L., is a volatile plant essential oil that is known to have insecticidal activity against
stored product pests such as rice weevil, Sitophilus oryzae (L.). Basil oil was diluted in acetone and applied to a
sponge held inside a tea strainer for fumigations in containers with and without rice. Basil oil fumigation
(3 ml, 10% concentration) caused high mortality in adult rice weevils when weevils were exposed in air in a
sealed 1 L plastic container. However, when basil oil was placed in packaged rice, weevil mortality was low
and reproduction was not affected. Effectiveness of plant essential oil fumigation should be conducted under
realistic conditions to avoid experimental artifacts and misleading results.
Published by Elsevier B.V. on behalf of Korean Society of Applied Entomology, Taiwan Entomological Society and
Malaysian Plant Protection Society.
Introduction
Rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae) is the
major pest of stored rice worldwide (Ahmed, 2001). Most domestic
and export markets have zero tolerance for grain contamination by
insects such as rice weevil. Fumigation treatment with phosphine or
other chemical fumigants ensures that grain can be stored and exported
in a clean and uninfested condition. Resistance to phosphine is a major
challenge to the control of rice weevil and other stored product pests
(Nayak et al., 2007), and alternative control methods are needed.
Adult rice weevils lay eggs on the rice grain and a single larva
hollows out an entire grain. Pupation occurs inside the grain and the
emerging adult bores its way to the outside (Shari and Mills, 1971).
The generation time from egg to adult is about 1 month (Howe,
1952). Adult weevils can live for 68 months or more, and a female
may lay 300400 eggs in her lifetime (Metcalf et al., 1962; Longstaff,
1981).
Several volatile plant essential oils have shown toxicity to stored
product pests (Bekele et al., 1997; Koul et al., 2008; Cardiet et al.,
2011; Regnault-Roger et al., 2012). Most studies have tested the effec-
tiveness of plant essential oils in Petri dishes or under other unrealistic
conditions. Basil oil, Ocimum basilicum L., was shown to have activity
against rice weevil in laboratory tests (Deshpande and Tipnis, 1977;
Popovi et al., 2006). We identied a toxic concentration of basil oil in
air under articial conditions, and then tested its toxicity to rice weevil
in commercial packages of rice.
Methods
Arice weevil colony was started froma heavily infested bag of whole
grain rice (Shirakiku Sukoyaka Genmai, Nishimoto Trading Co., Santa Fe
Springs, California, USA) obtained from a residence in Hilo, Hawaii. The
stock colony was maintained in multiple 3.8-liter plastic containers
with approximately 1 kg rice at 24 C (2 C) and a photoperiod of
12:12 (L:D) h (Davis and Dry, 1985). Adult weevils were periodically
moved to new containers with fresh rice. Gas chromatography of the
basil oil (Ocimum basilicum, 100% purity, Plant Therapy Essential Oils,
Twin Falls, ID) used in the experiments showed that it contained
70.6% methyl chavicol (also called estragole) and 21.2% linalool.
Mortality rate study
Pure basil oil was diluted in acetone to a concentration of 10%. A
preliminary study in Petri dishes suggested this concentration might
be effective in controlling rice weevil. A 3.0 mL aliquot of basil oil
solution was applied to a sponge (10 10 8 mm) and placed in a 2
mesh tea strainer (RSVP International, Inc., Seattle, WA). The acetone
was allowed to evaporate for 10 min (Cardiet et al., 2011). The tea
strainer with basil oil was suspended from the lid inside a 1 L plastic
container and 20 g rice (Diamond G brand, extra fancy Calrose Rice,
Sacramento, CA) and 35 adult weevils (24 weeks old) were added.
Weevil mortality was scored daily for six days. Weevils showing no
leg or antennal movement when prodded with a camel hair brush
were scored as dead. Acetone only was used as a control. A 1:1 sex
ratio was assumed (Howe, 1952). The basil oil treatment was replicated
six times and the control treatment four times in a single experiment.
Percentage mortality data were arcsin transformed and subjected to
analysis of variance and linear regression analysis (SAS Institute, 2010).
Journal of Asia-Pacic Entomology 17 (2014) 119121
Corresponding author at: USDA ARS, 64 Nowelo St., Hilo, Hawaii 96720.
E-mail address: Peter.follett@ars.usda.gov (P.A. Follett).
1226-8615/$ see front matter. Published by Elsevier B.V. on behalf of Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection Society.
http://dx.doi.org/10.1016/j.aspen.2013.11.008
Contents lists available at ScienceDirect
Journal of Asia-Pacic Entomology
j our nal homepage: www. el sevi er . com/ l ocat e/ j ape
Basil oil fumigation of packaged rice
An experiment was conducted to determine if the results from the
mortality rate study in an empty plastic container could be duplicated
in a commercial bag of rice. Basil oil was diluted in acetone to a concen-
tration of 10%, 3 ml was applied to a sponge, and the sponge was placed
in a tea strainer as described above. Acetone alone was used as a control.
The tea strainer with basil oil was positioned in the middle of a 2.26 kg
(5 lb) or 4.52 kg (10 lb) bag of rice (Diamond G brand, extra fancy
Calrose Rice, Sacramento, CA) and 50 adult rice weevils were added.
After three weeks, packages of rice were opened and all weevils were
removed and scored for mortality. The bags were then resealed and
held for an additional six weeks and opened to count any emerged F
1
adults. No emergence of F
1
adults would indicate that parental adults
were killed or sterilized, or that progeny did not develop in the rice
grains, whereas emergence of F
1
adults would indicate that the basil
oil treatment was not completely effective in preventing reproduction.
The basil oil treatments were replicated ve times for each bag size
(2.26 and 4.52 kg bags), and four untreated bags of rice of each size
served as controls. Percent mortality data were arcsin transformed
and subjected to analysis of variance and means separations were
calculated using a Tukey's test at alpha = 0.05. Untransformed data
for F1 progeny were analyzed similarly.
Dose response test in packaged rice
After results fromthe packaged rice study showed a limited effect of
10% basil oil on rice weevil mortality and reproduction, a study was
conducted to examine the effect of basil oil concentration of weevil
mortality. Basil oil concentrations of 0 (acetone only), 25%, 50%, and
100% (no acetone) were applied to sponges in tea strainers and placed
in 4.52 kg rice bags with 50 adult rice weevils. Bags were opened after
three weeks and weevil mortality was scored. Mortality data were
arcsin transformed and subjected to analysis of variance.
Results
Mortality rate study
The effect of day, basil oil treatment (3 ml, 10% concentration), and
the day * basil oil interaction on weevil mortality were all highly signif-
icant (P b 0.0001). After exposure to basil oil fumigation for ve or six
days, weevil survival was 2.9 and 1.7% respectively (Fig. 1). During the
same period survivorship in control weevils was N96%. The equation
describing the linear regression of percentage mortality against time
was y (% mortality) = 131.2 23.3 (days) (R
2
= 0.90). The linear
regression of control mortality against time was not signicantly
different from zero.
Basil oil fumigation of packaged rice
Although basil oil was highly effective in controlling rice weevil in
empty plastic containers, its effectiveness in commercial rice packages
of similar volume was limited. Adult weevil exposure to basil oil
(3 ml, 10% concentration) had no effect on mortality, regardless of
package size (2.26 or 4.52 kg bag) compared to the untreated control
(P = 0.6). Mean mortality after three weeks ranged from 2.6 to 5.3%
in rice packages containing basil oil, and from 2.5 to 3.3% in untreated
bags (Fig. 2). Basil oil and package size also had no effect on weevil
reproduction (P = 0.58). The pooled mean (SE) number of offspring
after six weeks was 145.4 (15.5) and 143.3 (13.6) in the basil oil
and control treatments, and 155.6 (14.1) and 133.2 (14.7) in the
2.26 and 4.52 kg packages of rice (Fig. 3).
Dose response test in packaged rice
Increasing the concentration of basil oil did not result in signicantly
higher weevil mortality (P = 0.5) (Fig. 4). The numerical trend was for
increasing mortality as the percentage concentration increased, but
when 100% pure basil oil was used mean mortality was still only 12.7%.
Discussion
Basil oil fumigation can cause high mortality in rice weevils as dem-
onstrated by the nearly complete mortality when weevils were exposed
in an empty plastic container. However, when basil oil was placed in
packaged rice, weevil mortality was low and reproduction was not
Days after Treatment
0 1 2 3 4 5 6 7
%

S
u
r
v
i
v
a
l
0
20
40
60
80
100
Control
Basil Oil
Fig. 1. Percentage mortality in rice weevil treated with 10% basil oil or left untreated.
Size of Rice Bag
2.26 kg 4.52 kg
%

M
o
r
t
a
l
i
t
y
0
2
4
6
8
Basil Oil
Control
Fig. 2. Percentage mortality inrice weevil after three weeks incommercial packages of rice
of two sizes treated with 10% basil oil.
Size of Rice Bag
2.26 kg 4.52 kg
N
u
m
b
e
r

o
f

O
f
f
s
p
r
i
n
g

0
20
40
60
80
100
120
140
160
180
200
Basil oil
Control
Fig. 3. Number of adult rice weevil F1 offspring after six weeks in commercial packages of
rice treated with 10% basil oil.
120 P.A. Follett et al. / Journal of Asia-Pacic Entomology 17 (2014) 119121
affected. These results suggest that the basil oil vapor may be adsorbed
on the rice (Ciesla and Ducom, 2008; Cardiet et al., 2011), or that
weevils nd less affected or unaffected pockets of air within the densely
packed rice grains. Our working model was that basil oil might be
incorporated into a polymer matrix product that would be inserted
into a package of rice or applied as an internal patch or coating to the
package itself. The polymer matrix would provide a controlled-release
of basil oil fumes that would kill rice weevil in the package and prevent
reproduction. The use of a treated sponge inside a tea strainer was an
attempt to simulate the function of a polymer matrix. We did not
examine contact toxicity of repellency of basil oil to rice weevil.
Popovi et al. (2006) studied the biological activity basil oil against
Sitophilus oryzae in laboratory experiments. Contact toxicity using treat-
ed lter paper increased with increasing concentration to a maximum
of 28.1% after 48 h with a 2% basil oil concentration. When 100 g
wheat was mixed with 3 ml of a 2% solution of basil oil and sealed in a
plastic bag, mortality was 53% after 4 weeks. The number of offspring
from 20 unsexed weevils per container (n = 6) was signicantly
reduced after 7 weeks froma mean (SE) of 208.6 (2.0) per contain-
er in untreated wheat to 14.5 (1.7) in the wheat treated with 2% basil
oil. Our results showed no effect on offspring production in the
commercial bags of rice, but we were testing the efcacy of basil oil
vapor rather than its contact toxicity. Also, we tested the effects of
basil oil using a larger volume of rice (2.26 kg or 4.52 kg) than
Popovi et al. (2006) (100 g), which may have diluted the toxic affect
due to increased adsorption.
We note that the volume of basil oil (3 ml) used during the experi-
ments was not varied throughout the study, and use of a higher volume
of the basil oil solution might have resulted in increased effectiveness.
Our study did not examine repellency or contact toxicity, and these
properties may be important to basil oil effectiveness in certain situa-
tions. Future studies of plant essential oil fumigation effectiveness
should be conducted under realistic conditions to avoid experimental
artifacts and misleading results. Basil oil may have potential as a syner-
gist when used in combination with other fumigants, or heat or irradia-
tion treatments to control stored product pests (Follett et al., 2013). In
addition to insects, basil oil has shown fungicidal activity and may be
useful to preserve high-moisture stored grain against toxigenic fungi
(Cardiet et al., 2011).
Acknowledgments
We are grateful to Justin Bisel, Brandi Antonio and Kirsten Snook
(USDA-ARS, Hilo, Hawaii) for their help in conducting the experiments,
and to Marisa Wall and Nicholas Manoukis (USDA-ARS, Hilo, Hawaii)
for their comments on an early draft of the paper. USDA is an equal
opportunity provider and employer.
References
Ahmed, M., 2001. Disinfestation of stored grains, pulses, dried fruits and nuts, and other
dried foods. In: Molins, R. (Ed.), Food Irradiation: Principles and Applications. John
Wiley & Sons, New York, pp. 77112.
Bekele, A.J., Obeng-Ofori, D., Hassanali, A., 1997. Evaluation of Ocimum kenyense as a
source of repellents, toxicants and protectants in storage against three major stored
product insect pests. J. of Appl. Entomol. 121, 169173.
Cardiet, G., Fizeau, b, Barreau, C., Fleurat-Lessard, F., 2011. Contact and fumigant tox-
icity of some essential oil constituents against a grain insect pest Sitophilus
oryzae and two fungi, Aspergillus westerdijkiae and Fusarium graminearum.
J. Pestic. Sci. 85, 351358.
Ciesla, Y., Ducom, P., 2008. Interests in the mixture ethyl formate/allylisothiocyanate for
the fumigation of infested wheat by the rice weevil: Sitophilus oryzae L. and the
granary weevil: S. granarius L. In Proceedings of 8th International Conference of
Controlled Atmosphere and Fumigation in Stored Products, Chengdu, pp. 688692.
Davis, R., Dry, R.E., 1985. Sitophilus granarius, Sitophilus oryzae and Sitophilus zeamais, In
Handbook of Insect Rearing, Amsterdam. Elsevier Science Publishers 287289.
Deshpande, R.S., Tipnis, H.P., 1977. Insecticidal activity of Ocimum basilicum Linn. Pestic.
11, 1112.
Follett, P.A., Snook, K., Brown, S., Bisel, J., Okamura, M., Antonio, B., Haruki, A., Jansen, A.,
2013. Irradiation quarantine treatment for control of Sitophilus oryzae (Coleoptera:
Curculionidae) in rice. J. Stored Prod. Res. 52, 6367.
Howe, R.W., 1952. The biology of the rice weevil, Calandra oryzae (L.). Annal. Appl. Biol. 2,
168180.
Koul, O., Walia, S., Dhaliwal, G.S., 2008. Essential oils as green pesticides: potential and
constraints. Biopesticides International 4, 6384.
Longstaff, B.C., 1981. Biology of the grain pest species of the genus Sitophilus (Coleoptera:
Curculionidae): a critical review. Prot. Ecol. 2, 83130.
Metcalf, C.L., Flint, W.P., Metcalf, R.L., 1962. Destructive and Useful Insects, 4th ed.
McGraw-Hill Book Co., New York.
Nayak, M.K., Collins, P.J., Pavic, H., 2007. Developing fumigation protocols to manage
strongly phosphine-resistant rice weevils, Sitophilus oryzae (L.). In: Donahaye,
E.J., Navarro, S., Bell, C., Jayas, D., Noyes, R., Phillips, T.W. (Eds.), Proc. Int. Conf.
Controlled Atmosphere and Fumigation in Stored Products Gold-Coast
Australia. FTIC Ltd. Publishing, Israel, pp. 267273 (August 813, 2007).
Popovi, Z., Kostic, M., Popovic, S., Skoric, S., 2006. Bioactivities of essential oils from basil
and sage to Sitophilus Oryzae L. Biotechnol. Biotechnol. Equip. 20 (1), 3640.
Regnault-Roger, C., Vincent, C., Arnason, J.T., 2012. Essential oils in insect control: low-risk
products in a high-stakes world. Annu. Rev. Entomol. 57, 405424.
SAS Institute, 2010. JMP 9.0.0 User's Guide.SAS Institute, Cary, NC.
Shari, S., Mills, R.B., 1971. Developmental activities and behavior of the rice weevil inside
wheat kernels. J. Econ. Entomol. 64, 11141118.
% Basil Oil
0 25 50 100
%

M
o
r
t
a
l
i
t
y
0
5
10
15
20
Fig. 4. Percentage mortality after three weeks in rice weevil treated with various concen-
tration of basil oil.
121 P.A. Follett et al. / Journal of Asia-Pacic Entomology 17 (2014) 119121