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Lau & Nieh - 2016 - Salt Preferences of Honey Bee Water Foragers
Lau & Nieh - 2016 - Salt Preferences of Honey Bee Water Foragers
132019
RESEARCH ARTICLE
790
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
0.8 Start-high
concentration order (Fig. 1); (2) 0%, 0.05%, 0.4%, 0.75%, 1.5%,
Start-low 6% and 10% of all salts for the full-range tests (Fig. 2A); and (3)
0.7 ranges individually tailored for bee PER responses to each salt
B
concentration: NaCl (0–3%), MgCl2 (0–6%), and KCl and
0.6 a
Na2HPO4 (0–1.5%, Fig. 2B). These concentrations are all much
lower than NaCl concentrations used as training punishment in
0.5
Mean PER
791
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
0.4 b 0.4
0.2 0.2 b
Mean PER
0 0
0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6
1 KCl 1
0.8 a 0.8
0.6 0.6
0.4 0.4
b
0.2 0.2
0 0
0 1 2 3 4 5 6 7 8 9 10 0 0.4 0.8 1.2 1.6
1 Na2HPO4 1
aa
0.8 0.8
0.6 0.6
0.4 0.4
0.2 b 0.2
0 0
0 1 2 3 4 5 6 7 8 9 10 0 0.4 0.8 1.2 1.6
diameter, and contained water that was subject to higher rates of (continuous fixed effect) on bee PER responses (bee and colony
evaporation because it was in a shallow vessel (a bird bath) or in a identity were random effects). To determine the significance of bee
small fountain that constantly recirculated water over a broad surface identity on PER responses to the full salt concentration ranges, we
area. More water samples were collected from urban and semi-urban also ran a model with bee identity as a fixed effect. We then tested
sites; however, 36% of our water samples came from natural water responses to each salt separately, using a one-way repeated measures
sources. ANOVA and Tukey’s honestly significant difference (HSD) tests to
make pairwise comparisons (concentration coded as an ordinal
Statistics fixed effect). These analyses used data gathered on bees tested with
We used repeated-measures ANOVA with a REML algorithm to the same concentrations of each salt. Finer scale salt preferences
determine the effect of concentration order on bee PER responses were analyzed with one-way repeated measures ANOVA models for
with salt concentration as a continuous fixed effect and bee identity each salt. We did not use an overall ANOVA on these data because
and colony as random effects. We used the same analysis method to different concentrations were used for different salts. We used JMP
determine the effect of salt type (fixed effect) and salt concentration v10.0 statistical software.
792
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
c
N
Na 0.93±0.09
0.08 b
0.06
Na (%)
0.04
a a a a a
0.02
0
b
Beach Mg 0.11±0.01
Swimming pool
Pond (artificial, small)
Pond (artificial, large) 0.008
Pond (natural)
a a a a a a
Major roads
Irrigation 0.006
Mg (%)
Stream
Natural areas
50 0.004
km
0.002
Fig. 3. Sources of water collected by honey bees. Samples were obtained
from the locations shown for analysis. 0
b
K 0.05±0.01
RESULTS
Effect of concentration order 0.008
We presented bees with a series of Na concentrations and measured
their PER responses. There was a significant effect of concentration 0.006 a a a a a a
K (%)
e)
l)
ol
h
NaCl were not significantly different from responses to 1.5% NaCl
al
io
al
ac
po
rg
ur
at
sm
re
Be
(Tukey HSD, P>0.05), matching bee preferences in our subsequent
la
at
ig
g
St
in
(n
l,
l,
Irr
ia
ia
m
nd
tests.
fic
fic
im
Po
rti
rti
Sw
(a
(a
nd
nd
Po
793
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
PER responses (Fig. 2B). There were significant overall effects of Table 1. Review of the published concentration ranges of salt ions that
concentration for NaCl (F8,392=3.42, P=0.0008), MgCl2 attracted or repulsed bees in honey and in water and nectar visited by
(F7,687=7.85, P<0.0001) and KCl (F6,228=3.37, P=0.0033). The bees
only significant pairwise differences (Tukey’s HSD test, P<0.05) Concentration (% w/w)
occurred for MgCl2, between 0.03% and 1.5% (higher PER for Salts Acceptance Rejection
1.5%, as in the full concentration range test). There was no
Na 0.00191,a nectar 0.00541,g nectar
significant effect of Na2HPO4 (F6,324=0.65, P=0.69) over the
0.00752 honey 0.00591,g honey
concentration range 0–1.5%. 0.00791 honey 0.01303,b nectar
0.00933,a nectar 0–0.05c water
Salt concentrations in water that bees collected 0.02264 nectar 0.585,d water
We analyzed water collected by bees from 36 sites in San Diego and 0.04 water 3.599,h water
Los Angeles Counties (Fig. 3) from June 2012 to June 2013. In all 0.05c water 17.5310,f water
analyses, internal control ICP-AES standards showed negligible 0.07 swimming pool
0.155,d water
levels of each ion (0.0 ppm). There was a significant effect of water 0.396,d water
type on the concentration of Na (F6,29=43.0, P<0.0001), Mg 0.587,e water
(F6,29=5.79, P=0.0005) and K (F6,29=9.08, P<0.0001), but not on 1.5c water
the concentration of phosphate (F6,29=1.67, P=0.16). As expected, 2.348,f water
seawater (0.93%) and swimming pool water (0.6%) contained Mg <0.00051,a nectar 0.00243,b nectar
higher sodium concentrations than all other water sources (Tukey 0.007 swimming pool 0.01881,g nectar
0.00902 honey 0.02051,g honey
HSD, P<0.05; Fig. 4). For Mg and K, only seawater had
0.012 water 0.955,d water
significantly elevated ion levels compared with all other water 0.01851 honey 10c water
sources (Tukey HSD, P<0.05; Fig. 4). 0.065,d water
1.5c water
DISCUSSION K 0.000759,g water 0.06593,a nectar
Honey bees are known to collect and, in some cases, prefer ‘dirty 0.005 swimming pool 0.15–0.7511,i nectar
0.03251,b honey 0.36–1.303,b nectar
water’ that contains salts (Butler, 1940). Understanding these
0.06 water 0.37681,g honey
preferences has practical importance because such water can contain 0.14802 honey 0.39461,g nectar
harmful xenobiotics such as pesticides. Using the classic PER assay, 0.215,d water 0.42–0.892,b nectar
we found differences in the responses of water foragers to the tested 0–1.5c water 0.53–0.6913,i nectar
salts. Overall, mean PER responses to NaCl and MgCl2 were 22% 1.665,d water
higher than for Na2HPO4 and KCl. In general, water foragers 10e water
exhibited the fewest responses to 10% salt concentrations. However, Phosphate 2–343 (×10−6) water 0.05111,g nectar
11–48 (×10−6) swimming pool 0.06521,g honey
water foragers demonstrated tolerance to a far wider range of Na 0–1.5c water 1.425,d water
concentrations than to other ions, perhaps explaining why honey 0.00191,a nectar 10c water
bees can collect seawater. The concentrations that elicited the most 0.00471,a honey
PER responses varied somewhat depending upon the range of Concentration ranges are from minimum to maximum. Swimming pool water
concentrations tested (Fig. 2). The optimal salt concentrations were has higher ion concentrations and is separately denoted. Data from the present
1.5% for NaCl and MgCl2, 0–1.5% for KCl and 0.4–0.75% for study are in bold.
1
Na2HPO4 (Fig. 2A). The analysis of finer salt concentration ranges Afik et al. (2006); 2McLellan (1975); 3Waller et al. (1972); 4Nicolson and
yielded similarly shaped PER curves (Fig. 2B), though fewer W.-Worswick (1990); 5Butler (1940); 6Hooper (1932); 7de Brito Sanchez et al.
significant pairwise differences because we did not include the (2014); 8de Brito Sanchez et al. (2005); 9Abramson et al. (2013); 10Bhagavan
and Smith (1997); 11Waller (1972); 12Waller et al. (1976); 13Hagler (1990).
generally aversive 10% concentration. a
Non-avocado and non-onion nectar; bonion nectar; cPER assay (this paper);
d
water feeder dishes; esalt solutions applied to bee tarsomeres; fPER assay;
Responses to each salt g
avocado nectar and honey; hchoice between NaCl and sucrose; isugar
Bees exhibited the most PER responses for intermediate solution with K added.
concentrations of sodium and magnesium, corresponding to the
794
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
795
RESEARCH ARTICLE Journal of Experimental Biology (2016) 219, 790-796 doi:10.1242/jeb.132019
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