MEDICAL ENTOMOLOGY

Boric Acid Bait Kills Adult Mosquitoes (Diptera: Culicidae)

RUI-DE XUE AND DONALD R. BARNARD
Center for Medical, Agricultural, and Veterinary Entomology, USDAÐARS, P.O. Box 14565, Gainesville, FL 32604

J. Econ. Entomol. 96(5): 1559Ð1562 (2003)

ABSTRACT The toxicity of boric acid solutions to adult Anopheles quadrimaculatus Say, Culex
nigripalpus Theobald, and Aedes albopictus Skuse was evaluated in the laboratory. Median lethal
concentrations (LC50 in %) at 24-h exposure for male and female An. quadrimaculatus were 0.317 and
0.885, respectively; for Cx. nigripalpus, 0.273 and 0.560, respectively; and for Ae. albopictus, 0.174 and
0.527, respectively. The LC50 values at 48-h exposure for male and female An. quadrimaculatus were
0.101 and 0.395, respectively; for Cx. nigripalpus, 0.098 and 0.255, respectively; and for Ae. albopictus,
0.078 and 0.244, respectively. In laboratory tests, access for 48 h to sucrose (10%) water containing
1% boric acid (boric acid bait) resulted in 98% mortality in blood fed, gravid, and parous Ae. albopictus.
When offered a choice between boric acid bait and sucrose water, 52% of male and 33% of female Ae.
albopictus ingested sufÞcient boric acid bait in 24 h to cause death; after 48 h, respective percent
mortalities were 88 and 58%. In outdoor tests, in a walk-in screened cage (156 m3) containing 1,250
female Ae. albopictus, mosquito biting rates on the exposed forearm of a human subject in 3-min
exposure were reduced ⱖ78% for the boric acid bait treatment, compared with a sucrose water control.

KEY WORDS toxicity, Aedes albopictus, Anopheles quadrimaculatus, Culex nigripalpus

CONVENTIONAL CONTROL OF ADULT mosquitoes is based
primarily on aerial and ground ultra-low volume ap-
plication of insecticides. But these techniques risk
environmental pollution because of the large areas
that are treated and the quantities of insecticides that
are applied. There is also the potential for develop-
ment of insecticide resistance in the mosquito popu-
lation. Each of these consequences compels us to Þnd
new ways to reduce the use of adulticides and to
develop safe and effective alternative control methods
for adult mosquitoes.
Because adult mosquitoes require carbohydrates
for energy, survival, and reproduction (Yuval 1992,
Foster 1995), one alternative to aerial/ground appli-
cation of insecticides is to provide mosquitoes a poison
mixed with their food. A bait formulation of boric acid
has been used in this way to control pest ants (Klotz
et al. 1997) but not to kill mosquitoes. Sucrose is an
excellent candidate for use in a bait product for mos-
quitoes because it is a component of the sugars they
acquire in nature from honeydew, plant sap, and nec-
tar (Schlein and Muller 1995, Burkett et al. 1999).
The study reported herein was performed to deter-
mine the potential usefulness of boric acid as a mos-
quito control agent. In devising it, we sought answers
to three questions. First, will a mixture of boric acid,
Written informed consent was obtained for all human subjects used
in this study in accordance with protocol IRB-01 #445-96 as currently
approved by the University of Florida, Health Sciences Center, In-
stitutional Review Board for Human Subjects. Approval to use animals
in this research has been granted by the Institutional Animal Care and
Use Committee at the University of Florida under project #A622.
sucrose, and water kill adult mosquitoes and, if so,
what is the lethal concentration? Second, does the
effectiveness of a bait solution vary with mosquito
species, sex, or the physiological status of the female?
Finally, will mortality in a mosquito population caused
by boric acid ingestion result in a reduction of the
mosquito biting rate on humans?
Boric acid baits could be used to reduce adult mos-
quito populations if they are determined to be lethal
to mosquitoes. In this regard, bait stations augmented
with a chemical feeding attractant and designed for
economy and portability would provide a safe, non-
pesticidal, point-source control technology for mos-
quitoes in urban and suburban environments and in
some types of livestock housing.
Materials and Methods
Mosquitoes. Anopheles quadrimaculatus Say, Culex
nigripalpus Theobald, and Aedes albopictus Skuse
were reared and maintained in the laboratory under
controlled temperature (27⬚C), relative humidity
(70%), and a photoperiod of 14:10 (L:D) h. Adults
emerged in stock cages (⬇500 cm3), were provided
10% sucrose in water ad libitum, and were blood fed
weekly on restrained 5Ð7-wk-old chicks or membrane
fed on bovine blood. Methods for rearing mosquito
larvae followed those described by Gerberg et al.
(1994).
Both sexes of each species were evaluated in acute
toxicity tests in the laboratory. We used Ae. albopictus
to determine the effect of boric acid on blood en-
1560 JOURNAL OF ECONOMIC ENTOMOLOGY
Table 1. Toxicity of boric acid baits to male and female Anopheles quadrimaculatus, Culex nigripalpus, and Aedes albopictus in the
laboratory after 24- and 48-h exposure
Species and sex 24 h
LC50 (%) 95% C.I.
Aedes albopictus
Male 0.174 0.146Ð0.199
Female 0.527 0.479Ð0.577
Culex nigripalpus
Male 0.273 0.117Ð0.397
Female 0.56 0.498Ð0.626
Anopheles quadrimaculatus
Male 0.317 0.289Ð0.344
Female 0.885 0.800Ð1.005
gorged, gravid, and parous females, whereas both male
and female Ae. albopictus were used in choice tests
between boric acid in sucrose water (i.e., boric acid
bait) and sucrose water alone. Nulliparous 5Ð7-d-old
Ae. albopictus were used to determine the effect of
boric acid bait on mosquito biting rates on a human
subject in an outdoor cage.
Acute Toxicity Tests. Treatments in toxicity bioas-
says comprised Þve concentrations of boric acid
(0.0001, 0.001, 0.01, 0.1, and 1%) dissolved in deionized
water containing 10% sucrose (i.e., sucrose water).
Ten percent sucrose in deionized water was used as
the control. Before each test, 50 ml of one of the Þve
treatments, or sucrose water control, was transferred
to a 60-ml plastic cup containing a single cotton ball.
A cup with either treatment or control solution was
then placed inside a mosquito cage (45 ⫻ 38 ⫻ 38 cm)
that held 100 male and 100 female mosquitoes. In
separate sets of bioassays that ended after 24 h and
after 48 h, the cups were removed from each cage and
the number of dead mosquitoes in the cage collected,
sexed, and their numbers recorded. Each toxicity bio-
assay was replicated four times for each mosquito
species.
Boric Acid Bait-Induced Mortality in Blood Fed,
Gravid, and Parous Aedes albopictus. In these tests,
the effect of boric acid (1%) bait with sucrose water
on mortality in blood fed, gravid, and parous Ae. al-
bopictus was compared. Blood fed mosquitoes were
obtained by feeding 7-d-old females on restrained
5Ð7-wk-old chicks and then testing them immediately
(blood fed), after holding for 3 d (with sucrose water
available ad libitum) (gravid), or after 7 d (with su-
crose water available ad libitum, and after oviposition)
(parous). Blood fed and gravid females were tested
using 25 mosquitoes per cage (45 ⫻ 38 ⫻ 38 cm), and
parous mosquitoes using 200 females per cage. Mor-
tality was assessed after 48 h using the methods de-
scribed above. Each test was replicated four times.
Choice Tests of Boric Acid Bait versus Sucrose Wa-
ter with Aedes albopictus. These tests were made to
determine the preference of male and female mos-
quitoes for boric acid (1%) bait or sucrose water when
both were simultaneously provided in a cage. Two
cages were used in each test, each containing 100 male
and 100 female, 5Ð7-d-old Ae. albopictus. One cage
Vol. 96, no. 5
Exposure time
48 h
Slope LC50 (%) 95% C.I. Slope
4.143 0.078 0.069Ð0.089 4.961
3.036 0.244 0.220Ð0.268 4.876
5.047 0.098 0.085Ð0.112 3.905
9.92 0.255 0.233Ð0.277 5.337
4.568 0.101 0.019Ð0.196 3.719
3.252 0.395 0.328Ð0.461 5.193
(treatment) received one cup each of boric acid bait
and sucrose water. The second cage (control) re-
ceived two cups of sucrose water. Choice was deter-
mined by assessing mortality in males and females in
each cage after 24 h. Choice tests were repeated four
times.
Effect of Boric Acid Treatment on Aedes albopictus
Biting Rates on Humans. An outdoor, walk-in,
screened cage (156 m3) was used for this study. A total
of 1,250 female Ae. albopictus was released into the
cage and 1 h later the number of mosquito bites in 3
min on the exposed forearm of a human volunteer was
recorded. Afterward, four 200-ml black plastic cups,
each containing 100 ml of boric acid (1%) bait, were
placed in each corner of the cage 1 m above ground
level on a wood rack. After 48 h, the same human
volunteer reentered the cage and recorded the num-
ber of mosquito bites received on an exposed forearm
in 3 min. The control scenario used the same regimen
with sucrose water substituted for the boric acid bait.
The same cage and human subject were used each
time to assess mosquito biting rates. Treatment and
control tests were each replicated six times.
Data Analysis. All tests were organized using a com-
pletely randomized design. Acute toxicity data were
analyzed using probit analysis (Finney 1971). Re-
sponse data for percent mortality in blood fed, gravid,
and parous mosquitoes, choice tests, and the mosquito
biting rates in outdoor cage tests were analyzed using
StudentÕs t-test or analysis of variance (SAS Institute
1988) with means separation in the latter case using
TukeyÕs honestly signiÞcant difference test.
Results
Acute Toxicity Tests. Boric acid baits were more
toxic to male than to female mosquitoes and were
more toxic to Ae. albopictus and Cx. nigripalpus than to
An. quadrimaculatus (Table 1). In general, LC50 values
decreased 50 Ð70% after 48-h exposure, compared with
24-h exposure. There was attenuation of toxicity (i.e.,
decreased slope of the Þtted regression line) after 48 h
for male An. quadrimaculatus and for Cx. nigripalpus,
with the opposite effect for female An. quadrimacu-
latus and Ae. albopictus.
October 2003 XUE AND BARNARD: BORIC ACID BAIT FOR MOSQUITOES
Table 2. Mean percent mortality (SE) in blood fed, gravid, and
parous Aedes albopictus after 48-h exposure to boric acid bait or
sucrose water in laboratory tests
Physiological
Boric acid baita Sucrose water
state
Blood fed 99.0 (1.7)a 1.0 (0.4)b
Gravid 99.0 (1.7)a 4.0 (2.8)b
Parous 98.0 (2.4)a 4.0 (2.6)b
a
Means in each row followed by same letter are not signiÞcantly
different (P ⫽ 0.05) using StudentÕs t-test (SAS 1988).
Boric Acid-Induced Mortality in Blood Fed, Gravid,
and Parous Aedes albopictus. Access to boric acid bait
signiÞcantly increased mortality in blood fed (t ⫽
84.47, df ⫽ 24, P ⬍ 0.001), gravid (t ⫽ 49.61, df ⫽ 24,
P ⬍ 0.001), and parous Ae. albopictus (t ⫽ 42.52, df ⫽
196, P ⬍ 0.001), compared with females that had access
to sucrose water only (Table 2). Percent mortality for
treatment and control groups was ⱖ98% and ⱕ4%,
respectively.
Choice Tests of Boric Acid Bait versus Sucrose Wa-
ter with Aedes albopictus. When presented with a
choice between boric acid bait and sucrose water, 52%
of males (t ⫽ 6.53, df ⫽ 99, P ⬍ 0.01) and 33% of
females (t ⫽ 5.68, df ⫽ 99, P ⬍ 0.05) fed at least once
from the boric acid bait cup and died in the Þrst 24-h
exposure (Table 3). After 48 h, mortality increased to
88% for males (t ⫽ 16.67, df ⫽ 99, P ⬍ 0.01) and 58%
for females (t ⫽ 8.85, df ⫽ 99, P ⬍ 0.01). Conversely,
12% of males and 42% of females did not ingest sufÞ-
cient boric acid bait in 48 h to induce mortality.
Effect of Boric Acid Treatment on Aedes albopictus
Biting Rates on Humans. Availability of boric acid bait
in the outdoor cage test signiÞcantly (F ⫽ 29.45, df ⫽
3,20, P ⬍ 0.001) reduced biting rates (ⱖ78%) by Ae.
albopictus on the human subject (Table 4). Biting rates
before introduction of boric acid bait or sucrose water
and 48 h after introduction of sucrose water were not
signiÞcantly different.
Discussion
Boric acid has been tested as a toxic bait for house
ßies and ants. In laboratory tests, LC50 values for boric
acid in 10% sugar water for adult house ßies, Musca
Table 3. Mean percent mortality (SE) of male and female Aedes
albopictus in choice tests after 24- and 48-h exposure to boric acid
bait and/or sucrose water in laboratory tests
Exposure period
Boric acid baita Sucrose water
and sex
24 h
Male 51.8 (11.3)a 3.5 (3.8)b
Female 33.5 (10.1)a 0.3 (0.4)b
48 h
Male 88.0 (2.4)a 7.5 (5.6)b
Female 58.0 (10.5)a 1.0 (0.7)b
Tests comprised the choice of boric acid bait or sucrose water in one
cage and sucrose water only in the second cage (see text for details).
a
Means in each row followed by same letter are not signiÞcantly
different (P ⫽ 0.05) using StudentÕs t-test (SAS 1988).
1561
Table 4. Number of bites in 3 min by Aedes albopictus on
exposed forearm of a human subject in an outdoor cage before and
after 48-h exposure of mosquitoes to boric acid bait or sucrose
water
Boric acid bait Sucrose water
Test
Before After 48 h Before After 48 h
1 1.8 0.2 2 1.2
2 2.2 0.2 1.8 1.4
3 2.3 0.5 1.9 1.4
4 2.2 0 1 1.4
5 1.9 0.2 2.3 1.7
6 1.2 0.2 2.6 1.8
Mean (SE)a 1.9 (0.4)a 0.2 (0.1)b 1.9 (0.5)a 1.5 (0.2)a
a
Means (as percent of 1,250 female Aedes albopictus) followed by
same letter are not signiÞcantly different (P ⫽ 0.05) using TukeyÕs
HSD test (SAS 1988).
domestic (L.), ranged from 0.37% to 0.88% (Hogsette
and Koehler 1992). For the red imported Þre ant,
Solenopsis invicta Buren, the LC50 decreased from
1.27% at 3 d, to 0.11% at 8 d (Klotz et al. 1997). The
results of our study show that boric acid bait has
toxicity to mosquitoes similar to house ßies and is more
toxic to mosquitoes than to Þre ants.
Our results also showed that blood fed, gravid, and
parous mosquitoes ingested sufÞcient boric acid bait
in 48 h to cause mortality. This observation is impor-
tant because it reveals that the toxicity of boric acid
baits to mosquitoes is unaffected by the physiological
state of the female.
Choice tests showed that boric acid bait was not
ingested at lethal levels in 24 h by 12 and 67% of male
and female mosquitoes, respectively; however, these
percentages were halved after 48 h of exposure. High
concentrations of boric acid are known to inhibit in-
gestion of baits by house ßies, cockroaches, and ants
(Hogsette and Koehler 1994, Strong et al. 1993, Klotz
et al. 1997). The reason for such inhibition is not
presently understood and the basis for repellency of
high concentrations of boric acid to mosquitoes needs
further study. One objective, in this regard, would be
to determine whether baits can be formulated to re-
duce boric acid concentration while maintaining or
improving efÞcacy and palatability to mosquitoes.
Use of boric acid bait may be an effective strategy
to control adult mosquitoes. Boric acid mixed with
sucrose and made available to mosquitoes in nature
could mitigate some of the safety and environmental
concerns associated with conventional mosquito adul-
ticides. Furthermore, it should be possible to formu-
late and deliver boric acid baits in the environment in
an effective way. Possible delivery systems include
various types of point-source bait containers, such as
hanging bait stations, or traps, that are augmented with
mosquito feeding attractants. Targeted spray applica-
tions of boric acid baits could also be made to vege-
tation (provided phytotoxicity is not a limiting factor)
or to inanimate surfaces, including those in mosquito
refugia.
1562 JOURNAL OF ECONOMIC ENTOMOLOGY
Acknowledgments
Pierre Larochelle and Johnny Jackson provided technical
support for this study.
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Received for publication 27 September 2002; accepted 3 June
2003.