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Toxicity and Repellency of Borate-Sucrose Water Baits to Argentine Ants (Hymenoptera: Formicidae)

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The oral toxicity of boron compounds to the Argentine ant, Linepithema humile (Mayr), was evaluated in laboratory tests. The ants were provided 25% sucrose water containing 0.5 and 1% boric acid, disodium octaborate tetrahydrate, and borax. Lethal times of these solutions were a function of the concentration of boron. In field tests, the ants showed no discrimination between disodium octaborate tetrahydrate and boric acid. There was a significant reduction in consumption of sucrose water with > 1% boric acid.
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HOUSEHOLD AND STRUCTURAL INSECTS
Toxicity and Repellency of Borate-Sucrose Water Baits to Argentine
Ants (Hymenoptera: Formicidae)
JOHN H. KLOTZ, LES GREENBERG, CHRISTOPHER AMRHEIN,
1
AND MICHAEL K. RUST
Department of Entomology, University of California, Riverside, CA 92521
J. Econ. Entomol. 93(4): 1256Ð1258 (2000)
ABSTRACT The oral toxicity of boron compounds to the Argentine ant, Linepithema humile
(Mayr), was evaluated in laboratory tests. The ants were provided 25% sucrose water containing 0.5
and 1% boric acid, disodium octaborate tetrahydrate, and borax. Lethal times of these solutions were
a function of the concentration of boron. In Þeld tests, the ants showed no discrimination between
disodium octaborate tetrahydrate and boric acid. There was a signiÞcant reduction in consumption
of sucrose water with 1% boric acid.
KEY WORDS Linepithema humile, ant baits, borates
BORON CONTAINING COMPOUNDS such as borax (sodium
tetraborate decahydrate) and boric acid have been
used since the early 1900s against ants (Rust 1986).
Our studies with sucrose water containing boric acid
for control of household ant pests have demonstrated
that at low concentrations (1%) boric acid is slow-
acting and nonrepellent thereby enhancing long-term
ingestion (Klotz and Moss 1996, Klotz et al. 1997). The
delayed activity of boric acid promotes a thorough
distribution of the active ingredient within the nest,
leading to death of the entire colony (Stringer et al.
1964, Klotz et al. 1996).
Commercial ant baits with boric acid or borax as an
active ingredient typically use concentrations of 5%.
For example, Niban (Nisus, Rockford, TN) and Bush-
whacker (Bethurum Research and Development,
Galveston, TX) granular baits use 5 and 18% boric acid,
respectively; and Terro Ant Killer II (Senoret Chem-
ical, Kirkwood, MO) liquid bait uses 5.4% borax. The
new liquid ant baits being developed use much lower
concentrations of boric acid. Drax Liquidator (Water-
bury Companies, Waterbury, CT), Dr. MossÕs Liquid
Bait System (JT Eaton, Twinsburg, OH), and Advance
Liquid Ant Bait (Whitmire Micro-Gen, St. Louis, MO)
use 1% boric acid in sucrose water. Another borate,
disodium octaborate tetrahydrate, has been formu-
lated at 1% in a sweet liquid bait, VG AB 3 (EPA
registration pending), for use against carpenter ants
(Wegner 1998).
Our two objectives in this study with Argentine ant,
Linepithema humile (Mayr), were to (1) compare the
oral toxicities of low concentrations of boric acid,
borax, and disodium octaborate tetrahydrate; and (2)
test for a feeding preference between disodium oc-
taborate tetrahydrate and boric acid and for different
concentrations of boric acid. Results of this study may
help optimize development of boron liquid bait for-
mulations for controlling ants.
Materials and Methods
Toxicity Tests. Argentine ants were collected from
a citrus grove on the Riverside campus, University of
California (Riverside County). The ants were pro-
vided water but no food for 1 d before bait exposure.
Concentration-mortality was determined with proce-
dures described by Klotz et al. (1998). Crystalline
boric acid and anhydrous borax (99% [AI]; Sigma, St.
Louis, MO), and TIM-BOR (98% disodium octaborate
tetrahydrate; U.S. Borax, Valencia, CA) were dissolved
in 25% (wt:vol) sucrose-deionized water solutions to
produce two concentrations (0.5 and 1%) of each
boron compound. Bait solutions were added to cotton
plugs daily inside small petri dishes with 10 ants. Treat-
ments and controls (25% sucrose-deionized water)
were replicated Þve times. The bait solutions were
available continuously to the ants for the duration of
the test. Daily observations on cumulative mortality
were recorded for 5 d.
Preference Tests. Binary choice tests were designed
to determine feeding preferences of the ants on dif-
ferent solutions. The consumption of solutions deliv-
ered to the ants in side-by-side feeding stations at-
tached to trees was used to measure preference.
Feeding stations were constructed from 50-ml capped
centrifuge tubes by drilling a 2-cm-diameter hole
through a 3.2-cm-diameter cap. A 4.5-cm square of
WeedBlock (Easy Gardener, Waco, TX), a porous
plastic material through which the ants could drink,
was centered over the top of the centrifuge tube Þlled
with solution and the cap screwed down to secure it
in place. The bait stations were inverted and taped at
eye level to the trunks of trees containing foraging
trails of Argentine ants. To test their preference be-
tween 1% disodium octaborate tetrahydrate and 1%
1
Department of Environmental Sciences, University of California,
Riverside, CA 92521.
0022-0493/00/1256Ð1258$02.00/0 2000 Entomological Society of America
boric acid (both in 25% sucrose water), tubes of each
solution were taped next to each other on each of 15
trees.
A similar test with 10 trees each was conducted for
sodium (Na
) and pH preference in solutions con-
taining equal concentrations of boron. When diso-
dium octaborate dissolves in water the following re-
action occurs:
Na
2
B
8
O
13
4 H
2
O
DSOBTH
9 H
2
O 3 2Na
2OH
8 H
3
BO
3boric acid
Because of the differences in molecular weight, a 1%
(10 g/liter) solution of disodium octaborate tetrahy-
drate is equivalent to a 1.2% boric acid solution with
1.9 g/liter of NaOH added. Boric acid solutions were
prepared with NaOH and NaCl, to equal the boron
and Na
content of 1% disodium octaborate tetrahy-
drate solutions. All solutions were prepared in 25%
sucrose water. The boric acid solution with NaCl (2.8
g/liter) was used to study the relative effects of Na
and pH preference. This solution had a pH that was
similar to the boric acid solution (4.7), but had the
same Na
concentration as the solution with NaOH.
The boric acid NaOH solution had a pH and Na
concentration that were similar to the 1% disodium
octaborate tetrahydrate (7.7).
To test for discrimination (preference test) of dif-
ferent concentrations of boric acid in sucrose water,
each boric acid solution (0.5, 1.0, 2.0, and 4.0%) was
paired with a control containing only sucrose water.
Each concentration was replicated on 10 trees. All test
solutions and controls used in these studies contained
25% sucrose in deionized water (wt:vol).
The ants were allowed to feed on the various solu-
tions for at least 24 h after which the amount con-
sumed by the ants in each of the vials was recorded.
Consumption was determined gravimetrically with 1%
boric acid and disodium octaborate tetrahydrate. Con-
sumption was determined volumetrically in the other
preference tests and standardized in milliliters per
hour by dividing the total volume consumed by the
number of hours the ants were allowed to feed.
Statistical Analysis. Mortality data were corrected
with AbbottÕs (1925) formula and analyzed by probit
analysis (Raymond 1985) to determine lethal times
(LT
50
) for each concentration. The value of chi square
was used to measure the goodness-of-Þt of the probit
regression line. Binary choice tests to determine feed-
ing preferences of ants for different solutions were
analyzed using a paired t-test comparison (StatView
1992). Regression analysis was performed on LT
50
Õs of
the different borates as a function of boron concen-
tration (Grapher 1996).
Results and Discussion
The time required to kill 50% of the workers of L.
humile (LT
50
) decreased with increasing concentra-
tion of the three boron compounds, indicated by non-
overlapping 95% CL (Table 1). Anhydrous borax, bo-
ric acid, and disodium octaborate tetrahydrate are
21.5, 17.5, and 21% boron by weight, respectively.
When the compounds are expressed in boron equiv-
alents, the LT
50
values are a function of the boron
concentration (Fig. 1). The log-linear relationship be-
tween LT
50
and boron concentration for the different
compounds was highly signiÞcant (P 0.01) with an
r
2
0.94.
Based on the consumption rates of 1% boric acid and
disodium octaborate tetrahydrate in sucrose water
Table 1. LT
50
values for L. humile workers fed boron compounds in 25% sucrose water
Treatment % concn LT
50
(95% CL), d Slope SE
No. of
ants
2
P
Boric acid 0.5 2.4 (2.1Ð2.6) 6.7 1.7 50 0.49 0.52
Boric acid 1.0 1.2 (1.0Ð1.4) 5.0 0.6 50 6.64 0.20
Disodium octaborate tetrahydrate 0.5 2.2 (1.8Ð2.9) 3.0 0.7 50 1.24 0.73
Disodium octaborate tetrahydrate 1.0 1.3 (1.1Ð1.5) 5.7 0.9 50 1.38 0.76
Borax 0.5 2.3 (2.1Ð2.6) 6.7 1.2 50 9.56 0.24
Borax 1.0 1.1 (0.9Ð1.3) 5.0 0.9 50 1.14 0.71
Fig. 1. Comparison of the relative toxicity of boric acid,
disodium octaborate tetrahydrate (DSOBTH), and anhy-
drous borax to Argentine ants. The relationship between
LT
50
and boron concentration was log-linear and highly sig-
niÞcant (P 0.01). [] Boric acid in 25% sugar water (this
study), [E] DSOBTH in 25% sugar water (this study), []
anhydrous borax in 25% sugar water (this study), [{] boric
acid in 25% sugar water (Klotz et al. 1998), [ƒ] boric acid in
10% sugar water (Klotz et al. 1998), Ñ log(Y) ⫽⫺0.83 *
log(X) 0.85 (r
2
0.94). Error bars are 95% CL.
August 2000 KLOTZ ET AL.: TOXICITY AND REPELLENCY OF BORATES TO ARGENTINE ANTS 1257
placed side-by-side in the binary choice test, we could
detect no preference (i.e., no signiÞcant difference)
by the ants for either solution (paired t-test, 1.22, df
14, P 0.24). When the pH is made approximately
equivalent for the two solutions by the addition of
NaOH, no preference was shown by the ants for either
solution (paired t-test, 1.46, df 9, P 0.18). Sim-
ilarly, no preference was shown for solutions with
different pH but equal Na
concentration when NaCl
was added to the boric acid solution (paired t-test,
0.96, df 9, P 0.36). Although there is no toxi-
cological difference or feeding preference between
boric acid and disodium octaborate tetrahydrate that
would make one better than the other, one advantage
to disodium octaborate tetrahydrate is that it dissolves
rapidly in water making it easier to formulate water
soluble baits. All solutions made from boric acid or
borate salts have in common the boron existing as
uncharged boric acid molecules at pH 8.
Consumption of sucrose water was signiÞcantly
higher than consumption of the 1% boric acid solu-
tions indicating unpalatibility of boric acid at 2 and 4%
(Table 2). Likewise, in a consumption test conducted
in the laboratory with Solenopsis invicta Buren, feed-
ing on solutions of 0.25, 1, and 5% boric acid in 10%
sucrose water there was a signiÞcant reduction in the
amount of the 5% solution consumed (Klotz et al.
1997). Therefore, to avoid repellency in liquid baits
with boron compounds as an active ingredient formu-
lations should use 1%.
Although there is little information available con-
cerning the physiological mode of action of borax and
boric acid on insects (Rust 1986), it has been shown
that borate ions form strong complexes with sugar
alcohols, such as inositol, and other organic functional
groups (Williams and Atalla 1981, Woods 1994, Hu et
al. 1997). In addition, boron may be involved in the
disruption of intercellular adhesion because saturated
boric acid solutions can be used to dissociate cells
(Goodrich 1942).
Acknowledgments
We thank the California Structural Pest Control Board
(Department of Consumer Affairs), University of Califor-
nia Statewide Integrated Pest Management Program, and
Citrus Research Board for their Þnancial support of this
research.
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Table 2. Consumption rate (ml/h) of Argentine ants feeding on
25% sucrose water and 25% sucrose water boric acid solutions
when ants were given a choice (10 trees per treatment)
Concn of boric
acid, %
a
Treated
(mean
b
SE)
Control
(mean
b
SE)
0.5 0.123 .019a 0.085 .016a
1 0.255 .034a 0.297 .059a
2 0.059 .009a 0.143 .018b
4 0.017 .008a 0.236 .040b
Means within a row followed by the same letter are not signiÞcantly
different (P 0.05, paired t-test comparison [StatView 1992]). NS,
not signiÞcant.
a
%, wt:vol.
1258 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 93, no. 4
... viscosity; Josens et al., 2017). In fact, boron baits are often consumed less than plain sucrose solutions in field conditions (Klotz et al., 2000;Ripa et al., 1999). ...
... Few studies investigate individual behaviour in the consumption of toxic bait (Hooper-Bùi & Rust, 2001;Silverman & Roulston, 2001), while most research was carried out with ant groups and colonies in laboratory or in the field (Baker et al., 1985;Greenberg et al., 2006). Moreover, bait consumption is typically measured several hours after bait access has been provided (Klotz et al., 2000;Ripa et al., 1999). ...
... We used 20% w/w as the control solution in each assay since it is included in the preferred range of concentrations for both species; however, we compare them taking into account that there are differences between optimal concentrations of each one (Josens et al., 1998). On the other hand, even when the motivation to sucrose might be similar, the non-toxic compounds added to the solutions may asymmetrically modify motivation due to the different nutritional requirements (Kaspari, 2020;Klotz et al., 2000). All in all, Argentine ants and Camponotus ants have shown clear differences in bait palatability preference (rejecting only boric acid in one species and only borax in the other one), and this was the starting point that we took as validation for our study. ...
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Managing household pests Advances in urban pest management StatView 4.5. 1992. StatView userÕs guide. Abacus Concepts Im-ported Þre ant toxic bait studies: evaluation of toxicants
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Rust, M. K. 1986. Managing household pests. In G. W. Ben-nett and J. M. Owens [eds.], Advances in urban pest management. Van Nostrand Reinhold, New York. StatView 4.5. 1992. StatView userÕs guide. Abacus Concepts, Berkeley, CA. Stringer, C. E., C. S. Lofgren, and F. J. Bartlett. 1964. Im-ported Þre ant toxic bait studies: evaluation of toxicants. J. Econ. Entomol. 57: 941Ð945.