Pest Management Science Pest Manag Sci (2007)

Residue distribution of the

acaricide coumaphos in honey following
application of a new slow-release formulation
Emmanuel Karazafiris,1 Chrysoula Tananaki,1 Urania Menkissoglu-Spiroudi2 and
Andreas Thrasyvoulou1
1 Laboratory of Apiculture-Sericulture, School of Agriculture, PO Box 251, Aristotle University of Thessaloniki, 54 124 Thessaloniki, Greece
2 Laboratory of Pesticide Science, School of Agriculture, PO Box 251, Aristotle University of Thessaloniki, 54 124 Thessaloniki, Greece

Abstract: Acaricide used in beehives for the control of varroa often leaves residues in bee products. The behaviour
and distribution of the acaricide coumaphos in honey following the application of a new slow-release strip
formulation (CheckMite+) was assessed. The bee colonies were allowed to build new combs without foundation,
and two strips were hung in the brood chamber of each colony for a period of 42 days. The distribution of
coumaphos residues in honey in relation to the position of the frame and the duration of treatment was examined
by collecting samples from each comb at various time intervals up to 145 days after treatment. In the brood
chamber, coumaphos was incorporated into honey from the first day of application, and residues accumulated
mainly in combs placed next to strips. In the adjacent combs, residues remained at low concentrations with slight
variations. In the honey chamber, residue concentrations on the day of strip removal ranged between 0.006 and
0.020 mg kg1 , while 79 days after application the concentration of coumaphos residues was below 0.020 mg kg1 .
Residues above the EC fixed maximum residue limit (MRL) of 0.1 mg kg1 were measured only in brood chamber
honey obtained from those combs placed next to strips. In these samples, 0.0600.111 mg kg1 of coumaphos was
detected up to 103 days after strip removal. Coumaphos residues in honey extracted from combs that were placed
at the edge of the brood chamber were found below the MRL value, even during the 42 day period of CheckMite+
strip treatment.
2007 Society of Chemical Industry

Keywords: CheckMite+; coumaphos; distribution; honey; residues

1 INTRODUCTION was granted a limited use permit in 1998 in the

One of the major problems in apiculture is the parasitic
mite Varroa destructor (Anderson & Trueman), which,
unless properly treated, seriously damages honey bee
colonies (Apis mellifera L.) worldwide. In Greece,
varroatosis was first detected in 1978,1 and since
then varroa mites have spread rapidly throughout
the country, causing heavy losses in apiculture.
Although great efforts have been made to find
effective pesticides for the control of this mite
infestation of the honey bee, few control agents are
available to the beekeeping industry. One of the first
agents used against the varroa mite in Greece was
the organophosphate insecticide coumaphos, active
ingredient of Asuntol-50 and Perizin.2 Coumaphos,
along with the non-authorized malathion, became the
most widely used chemicals against the varroa mite in
Greece during the 19791988 period. Since then, the
pyrethroid tau-fluvalinate has been used worldwide
by beekeepers to prevent varroatosis. The extensive
use of fluvalinate resulted in resistance developing in
Europe and the United States.3 5 In order to deal with
this, a slow-release strip formulation of coumaphos
United States and Canada. The liquid formulation
of coumaphos (Perizin) has already been used in
Europe. Moreover, in Greece extensive use of Perizin
started in 1999, when the resistance of the varroa
mite to tau-fluvalinate developed.4 After only 3 years
of coumaphos strip use in the USA, coumaphos-
resistant mites appeared.5 Mite resistance to acaricides
is becoming all too common in many parts of the
world. Varroa destructor populations resistant to both
fluvalinate and coumaphos are currently found in the
USA, and few mite control alternatives are on the
horizon.
Pesticide applications in beehives produce unde-
sirable residues in bee products. Several publications
report on acaricide residues in honey and other bee-
hive products.6 9 The levels of residues determined in
bee products depend on the amount of active ingre-
dient used, its chemical properties, the application
technique, the duration of the treatment, the sea-
son and the activity of the bees. Extended exposure
time or unregistered treatments result in high residue
concentrations and should be avoided.

Correspondence to: Urania Menkissoglu-Spiroudi, Laboratory of Pesticide Science, School of Agriculture, PO Box 251, Aristotle University of Thessaloniki,
54 124 Thessaloniki, Greece
E-mail: rmenkis@agro.auth.gr
(Received 3 March 2006; revised version received 11 August 2006; accepted 12 December 2006)
DOI: 10.1002/ps.1493

2007 Society of Chemical Industry. Pest Manag Sci 1526498X/2007/$30.00

E Karazafiris et al.
Coumaphos residues in Greek honey have been
monitored for several years,10 12 and model exper-
iments have been used to study the transfer of
coumaphos from contaminated beeswax to honey.13,14
All these studies report increased coumaphos diffusion
into honey and sugar feed when the concentration
of the acaricide in wax is high. According to these
studies, among all acaricides studied (coumaphos, tau-
fluvalinate, flumethrin, bromoproplate), coumaphos
has the greatest potential to contaminate honey, as it
is the least lipophilic substance studied and has the
highest tendency to diffuse out of wax into honey.7
A recent study of coumaphos distribution in the hive
ecosystem, conducted in experimental hives treated
with Perizin, the commercial liquid formulation of
coumaphos, showed diffuse contamination.15
Bayer recently offered coumaphos in a new
formulation under the name CheckMite+ in the
form of plastic strips (1.4 g active ingredient per strip)
which are hung in the brood chamber. This slow-
release formulation is used in the United States and
Canada under a limited use permit, and a registration
in Greece was also granted in 2006. CheckMite+
strips release sufficient active ingredient to control V.
destructor and the small hive beetle (Aethina tumida
Murray). According to the manufacturer, two strips
should be placed into the colony for 4245 days once
or twice per year. This new formulation facilitates
the treatment against varroa, as no supplementary
measures are needed and no liquid is sprayed in hives
during the cold months.
As in the case of other chemicals applied in
bee colonies, acaricide residues are expected to be
incorporated into honey and beeswax. The main safety
rule to decrease the risk of honey contamination is not
to collect the product during treatment. Bayer not
only recommends avoiding collecting honey during
the 42 days of treatment with CheckMite+ strips
but also suggests taking out honey supers before
application and replacing them 14 days after removal
of the strips. Some beekeepers, and especially those
with a great number of bee colonies, will not readily
accept removing the honey supers for 56 days. A few
of them may disregard the crucial suggestions and
harvest honey from hives that are under treatment.
In addition, one of the basic beekeeping procedures
is to move honey out of the brood nest, to relieve
congestion and put it where it can later be harvested.
Moving brood combs around while using the Demaree
system of swarm control is another common procedure
for beekeepers. It is thus imperative to study the
distribution of coumaphos residues in each separate
comb of the treated colony and relate the level of honey
contamination to a strips position and of course to
the duration of treatment and the time lapsed since
treatment. Tremolada et al.15 studied coumaphos
distribution in a hive ecosystem treated with Perizin
solution sprinkled on the top of hives, and found
increased coumaphos residues 1 month after treatment
and also diffuse contamination. The distribution of
coumaphos residues in the hive after application
of CheckMite+ strips is expected to be different
from that reported by Tremolada et al.15 because of
the difference in mode of application (CheckMite+
is a controlled-release formulation), the variability
of the distance and the space between each frame
and the strips. Heukamp16 reported that coumaphos
residues in brood chamber honey increased during the
application of CheckMite+ strips.
Several studies have shown that the technique used
to apply acaricides in beehives, the duration of the
treatment, the season and bee activity can all affect
residue levels in bee products.17 In the literature only
one study reports on coumaphos residues released
after CheckMite+ strip application.16
The goal of the present research was (i) to study the
distribution of coumaphos residues in each separate
comb of the hive after acaricide had been applied in a
slow- release formulation; (ii) to correlate the level
of honey contamination with coumaphos residues
to distance from strips, duration of treatment and
time lapsed after removal of strips. Honey samples
from each separate comb of the treated colonies
were collected and analysed to determine coumaphos
residues during the 42 days of treatment and for up
to 103 days after removal of strips. Honey chamber
residues were also assessed.
2 MATERIALS AND METHODS
2.1 Bee colonies and treatments
Four colonies that had not been treated against varroa
mites for the previous 12 months were used. All the
combs were removed and the bees were allowed to
build new combs without foundation. To facilitate
the building, colonies had been fed on syrup (1:1)
for 2 weeks. The experimental colonies occupied 20
frames of population, 68 frames of brood and were
in hives with two-chamber Langstroth bodies.
Two CheckMite+ strips (Bayer, Germany) were
placed in three colonies for 42 days. The strips were
hung between the 3rd and 4th and the 7th and 8th
frames of the brood chamber. The fourth colony was
the control where no chemicals were applied.
2.2 Sampling
Honey samples from three treated hives were collected
from each of the ten combs of the brood chamber on
days 0, 1, 3, 7, 14, 21, 28, 35, 42 (day of strip removal),
56, 70, 84, 98 and 145. Samples from each comb of
the control were collected on days 0, 42 and 145. The
sampling on day 0 was done immediately before the
application of CheckMite+ strips. In order to examine
the variability of coumaphos residues in honey within
a comb, honey was collected and analysed from five
different locations on each of two combs from each
colony. This preliminary experiment showed that
coumaphos residues are unevenly distributed within
the comb. For this reason, each honey sample of
this experiment was collected in the following way:
Pest Manag Sci (2007)
DOI: 10.1002/ps

35 pieces of comb were cut with a sharp knife
from different areas of the examined comb. Honey
was obtained from these pieces of comb by dripping
through a fine mesh in the laboratory. Special care was
taken not to press the sample and thus release wax
pieces into the honey.
Samples from the honey chamber were collected in
a similar way on days 0, 7, 16, 29, 42 and 79.
2.3 Chemical analysis
2.3.1 Reagents and chemicals
Ethanol, hexane, isooctane and water, purchased from
Merck Company (Germany), were of residue analysis
grade. Disposable RP-C18 solid-phase extraction
cartridges (BondElut, 500 mg) were obtained from
Varian (USA). The coumaphos certified reference
standard 99.9%, supplied by Bayer AG Animal
Health, Germany, was dissolved in acetone (1 mg
mL1 ) to obtain the stock standard solution, which
was stored at 18 C in the dark. Working standard
solutions were prepared by proper dilution of the
stock solution with matrix extract. For the purpose of
sample fortification, coumaphos solutions in acetone
were used.
2.3.2 Instrumentation
A Varian SPE-20 vacuum manifold was used for solid-
phase extraction. A 6890 Hewlett-Packard (USA) gas
chromatograph equipped with an HP 7683 autosam-
pler, a cool on-column injector and a microelectron
capture detector (ECD) was used. The HP Chem-
station software was used for instrument control and
data acquisition. A fused-silica capillary column HP-
608, 30 m 0.53 ID, 0.5 m film thickness, connected
to a deactivated fused-silica uncoated precolumn
(0.5 m 0.53 mm), was used. The column temper-
ature was initially 40 C, held for 0 min, and then
programmed up to 250 C, using two gradients: at
25 C min1 up to 200 C, where it was held for 1 min,
and at 5 C min1 up to 250 C, where it was held for
14 min. The injector was set at 40 C initially and
then it followed the column temperature programme
(oven temperature +3 C, track oven mode), while the
detector was set at 300 C. The carrier gas was helium
and the flowrate was 6 mL min1 . The make-up gas
was nitrogen at 60 mL min1 . The injected volume
was 2 L. Under these conditions, the retention time
of coumaphos was 23.609 0.012 min. Quantifica-
tion was done by the external standard method, and
the calibration curve was matrix matched.
2.3.3 Chemical measurements
A modified analytical method involving the extraction
of coumaphos from honey using a solid-phase
extraction procedure followed by GC determination
was used for the analysis of coumaphos residues in
honey.6 The method employed is simple, fast, reliable
and accurate.
Pest Manag Sci (2007)
DOI: 10.1002/ps
Residues in honey from slow-release coumaphos formulation
2.3.4 Sample preparation
A sample (5 g) was dissolved in ethanol + water (1 + 1
by volume; 10 mL) mixture in a screw-stoppered vial
with heating (40 C, 10 min). The honey solution
was passed under constant vacuum through a C18
cartridge that had been previously activated by washing
with ethanol (3 mL), followed by water (3 mL). The
cartridge was then washed with ethanol + water
(5 + 95 by volume; 3 mL) and finally dried under
vacuum for 15 min. Coumaphos was eluted with ethyl
acetate (2 mL) and hexane (2 mL). The solvent was
evaporated to dryness under vacuum and the residue
was redissolved in isooctane (2 mL), thus being ready
for GC/ECD analysis.
2.3.5 Recovery study
The recovery study was carried out by fortifying
untreated samples collected from the control hive
(5 g) with coumaphos standard solutions in acetone
(50 L). The method was assessed at three fortification
levels (5, 100 and 500 g kg1 ). After evaporation of
the solvent, the samples were treated as described
above and analysed. Five replicates of each recovery
assay and five unfortified blank samples were analysed.
The sensitivity of the method and the limit of detection
were estimated according to Thier and Zeumer.18
Interday and intraday repeatabilities were estimated by
analysing ten samples of matrix (990 L) fortified with
10 L of coumaphos standard solution (10 g mL1 ).
Regression analysis of the data obtained by running a
series of working solutions containing 5500 ng mL1
of coumaphos showed the ECD detector response to
be linear in the range 101000 pg, with a regression
coefficient of 0.9996. The calibration equation was y =
125567x 364.55. The recovery data are summarized
in Table 1. The relationship between added and
found amount was described by the linear regression
y = 1.0794x + 0.002, with R2 = 0.9999. According to
the above regression, the sensitivity of the method
was 107.94%. The limit of detection was calculated
on the basis of standard deviations of blanks and
samples at the lowest fortification level (0.005 mg
kg1 ), with f = 10 at the 95% confidence level.18
The estimated limit of detection was 0.0004 mg kg1 ,
and the limit of quantification was 0.005 mg kg1 ,
which was the lowest fortification level with acceptable
recovery. The interday and intraday repeatabilities
were respectively 0.1069 0.0009 with a coefficient
of variation of 0.86% and 0.0913 0.0016 with a
coefficient of variation of 1.76%.
Table 1. Mean recoveriesa of coumaphos from honey at various
fortification levels
Fortification level Accuracy Related standard Recovery
(mg kg1 ) (mg kg1 ) deviation (%) (%)
0.005 0.0047 3.04 93
0.1 0.0885 2.77 88
0.5 0.4618 0.95 92
a
Five replicates at each fortification level.
E Karazafiris et al.

3 RESULTS
3.1 Distribution of coumaphos within the comb
The distribution of coumaphos residues in honey
samples obtained from five different parts of two
combs is presented in Table 2. Great variation was
observed among coumaphos residue levels in honey
samples collected from different parts of the same
comb. The coefficients of variation were 28.02 and
32.11% for the two combs, indicating the uneven
distribution of residues within the comb. As many
portions as possible were collected from each comb
in order to obtain more representative samples. Most
of the brood comb honey samples came from three
pieces of the upper part of each frame, while samples
from combs in the honey chamber derived from five
positions.
3.2 Residues in the brood comb chamber
Data presented in Table 3 show the distribution of
coumaphos residues in the honey of each frame of
the brood chamber in relation to the position and
distance of the comb from the strips and the duration
of treatment of the three colonies. The values are the
mean values from the three hives. All samples collected
Table 2. The distribution of coumaphos residues in honey within a
comb
Concentration of coumaphos
(mg kg1 )
Position of sampling Comb No. 1
Upper left 0.076
Upper right 0.085
Centre 0.083
Down left 0.137
Down right 0.077
Average 0.092
Standard deviation 0.0257
Coefficient of variation (%) 28.02
from the control colony during the trial were free of
coumaphos residues (<LOQ = 0.005 mg kg1 ). Also,
residues were not detected 1 day before treatment,
which is an indication that bees and honey were free
from the acaricide. A dash in some dates indicates
that no analysis was made because no honey was
found in the respective comb. However, bees built
new combs to cover the gaps left by the removal of
the samples. The authors did try to avoid these new
combs, but sometimes it was impossible, especially
in combs with brood where the stored honey was
less. The variation of values shown in Table 3 could
be explained by the new wax that bees produce in
order to cover the gap, the presence of sealed or
unsealed honey, the presence of pollen and other
factors that could not be controlled. Residues were
incorporated into honey very rapidly and, by the third
day of application, residue concentrations measured
in two of the colonies were 0.101 and 0.162 mg kg1 ,
exceeding the maximum residue limit (MRL) of
0.1 mg kg1 .19 As the days of treatment proceeded,
residues accumulated in combs and coumaphos
concentration increased. At the end of the treatment,
the mean coumaphos concentrations in the three hives
were found to be 0.103, 0.161 and 0.133 mg kg1 .
Although differences between the three colonies
exist, the mode of residue distribution in combs is
similar. Figure 1 presents the average concentration
of coumaphos residues, considering the frames as
three groups. Concentrations measured in honey from
combs next to acaricide strips frames 3 and 4 and
Comb no. 2
frames 7 and 8 were higher and ranged from 0.097
0.042 to 0.289 mg kg1 . Residues in honey samples from
0.081 the adjacent combs frames 1 and 2, frames 5 and
0.107 6 and frames 9 and 10 were at lower levels and
0.105 ranged from 0.052 to 0.082 mg kg1 . The maximum
0.109 average coumaphos concentration of 0.131 mg kg1
0.089
was measured 35 days after treatment, while the
0.0285
32.11
maximum concentration of 0.289 mg kg1 was found
in the comb at position 8 on the last day of treatment

Table 3. Residues of coumaphos (mg kg1 ) in brood chamber honey during the 42 days of application and up to 103 days after the removal of
CheckMite+ strips. Average from three coloniesa

Day of honey collection

Position of frame 0 0 + 1 0 + 3 0 + 7 0 + 14 0 + 21 0 + 28 0 + 35 0 + 42 0 + 56 0 + 70 0 + 84 0 + 98 0 + 145

1 n.d.b 0.031 0.068 0.072 0.029 0.039 0.036 0.037 0.060 0.019 0.012
2 n.d. 0.009 0.045 0.085 0.081 0.062 0.077 0.075 0.076 0.066 0.055 0.062 0.065 0.066
3a n.d. 0.040 0.048
0.122 0.101 0.155 0.209 0.195 0.186 0.141 0.090 0.074 0.064 0.111
4 n.d. 0.047 0.072
0.133 0.142 0.130 0.138 0.198 0.160 0.095 0.095 0.076 0.088 0.082
5 n.d. 0.040 0.063 0.112 0.057 0.066 0.085 0.101 0.050 0.051 0.050 0.047 0.037
6 n.d. 0.063 0.062 0.063 0.065 0.065 0.067 0.023 0.031 0.039 0.030 0.043
7 n.d. 0.015 0.080 0.107 0.103 0.112 0.135 0.184 0.115 0.096 0.055 0.073 0.046 0.060
8 n.d. 0.162 0.226 0.126 0.177 0.108 0.232 0.229 0.105 0.130 0.085 0.048 0.071
9 n.d. 0.047 0.068 0.053 0.057 0.081 0.058 0.056 0.013 0.048 0.048 0.030 0.043
10 n.d. 0.114 0.101 0.100 0.048 0.042 0.084 0.091 0.077 0.029 0.049 0.025 0.040
Total n.d. 0.175 0.281 0.417 0.587 0.636 0.736 1.133 0.959 0.445 0.395 0.431 0.324 0.463
Mean n.d. 0.043 0.074 0.130 0.090 0.090 0.105 0.131 0.129 0.073 0.053 0.059 0.045 0.058
a
Bold represents the combs that were next to CheckMite+ strips during the application.
b
n.d. = not detectable.

Pest Manag Sci (2007)

DOI: 10.1002/ps
 Residues in honey from slow-release coumaphos formulation

0.25 frames 1+2+9+10 0 Table 4. Residues of coumaphos (mg kg1 ) in honey chamber during
frames 3+4+7+8 0 the 42 days of application and up to 37 days after the removal of
residues mg kg-1

0.2 frames 5+6 0 CheckMite+ strips

0.15 Number Day of sampling

MRL of comb
0.1
and hivea 0 0 + 1 0 + 7 0 + 16 0 + 29 0 + 42 0 + 79
0.05
1A n.d.b n.d. n.d. n.d. n.d. n.d. n.d.
0 4Ac n.d. n.d. n.d. n.d. n.d. 0.010 0.009
1
3
7
14

0+ 1
0+ 8
0+ 5
0+ 2
0+ 6
0+ 0
0+ 4
0+ 8
5
7A n.d. n.d. n.d. n.d. n.d. 0.013 0.014
0+
0+
0+

2
2
3
4
5
7
8
9
14
0+
0+

10A n.d. n.d. n.d. n.d. n.d. 0.006 0.006

days after treatment
1B n.d. n.d. n.d. n.d. n.d. 0.015 0.006
4B n.d. n.d. n.d. n.d. 0.006 0.012 0.013
Figure 1. Distribution of coumaphos residues in brood chamber;
7B n.d. n.d. n.d. n.d. 0.006 0.020 0.013
frames are divided in three groups according to their distance from
CheckMite+ strips. Numbers are mean values of three colonies. 10B n.d. n.d. n.d. n.d. 0.007 0.006 0.007
1C n.d. n.d. n.d. n.d. n.d. n.d. n.d.
4C n.d. n.d. n.d. n.d. 0.010 0.018 0.005
(day 0 + 42). The average coumaphos concentration 7C n.d. n.d. n.d. n.d. 0.011 0.016 0.007
14 days after the removal of strips was reduced to 10C n.d. 0.005 0.005 n.d. n.d. 0.018 n.d.
0.073 mg kg1 and remained rather constant till the a
The number and the letter in column 1 indicate the position of the
end of the trial. Even 103 days after the removal of frame and the hive respectively.
the strips, coumaphos concentrations of 0.112 and b
n.d. = not detectable.
0.111 mg kg1 were found in honey from two colonies. c Letters in bold represent the combs that were on top of the

CheckMite+ strips during the application.

Both samples were collected from combs at position 3
next to the CheckMite+ strip.
3.3 Residues in the honey chamber
Samples from the honey chamber were collected from
four combs of each hive, and the results of residue
analysis are presented in Table 4. The position of
frames 4 and 7 was on the top of the CheckMite+
strips hung in the brood chamber underneath. In
the honey super no detectable coumaphos residues
were found until the last application day. Coumaphos
residues in samples on the last day of treatment (day
0 + 42) ranged between 0.006 and 0.020 mg kg1 . The
concentration of coumaphos residues on day 0 + 79,
i.e. 37 days after removal of the strips, was below
0.02 mg kg1 .
4 DISCUSSION
The behaviour and distribution of the acaricide
coumaphos released from CheckMite+ strips in
treated colonies was assessed. Applications of strips
according to recommendation resulted in residues
below 0.1 mg kg1 . However, these experiments
show that a number of precautions should be
considered when assessing coumaphos residues in
honey. Coumaphos the active ingredient of the
strip formulation is slowly released and dispersed
throughout the colony by the legs and bodies of
bees. The distribution is uneven and depends on
the activity of the bee colony and mainly on the
proximity of combs to the acaricide strips. This
variability was verified from the analysis of individual
honey samples obtained from different parts of the
same comb. For this reason, and in order to obtain
more accurate results, 35 parts of a comb were
combined to give representative samples. It must be
noted that collection of representative samples for
residue analysis during a trial in beehives is a point of
serious concern. Variability in residue concentrations
of honey or wax samples between the combs of a
hive or even within a single comb has been reported
for fluvalinate applied as Apistan strips.17 Bogdanov
et al.6 have also stressed the need for representative
samples for honey residue analysis. The experimental
results of this study showed considerable variability
in residue levels in brood chamber honey derived
from each of the ten combs of a hive. During the
42 days the strips remained in the hives, coumaphos
residues accumulated and their concentration in brood
honey rose to an average of 0.129 mg kg1 in the
three hives on the last day (0 + 42) of treatment
(Table 3). After removal of the strips, coumaphos
residues declined, but they were still detectable on
the last day (0 + 145). Data presented in Table 3 and
Fig. 1 indicate considerable contamination of honey
obtained from combs near the strips. Specifically,
on day 0 + 42, residue levels in honey samples
from combs in contact with the strips exceeded the
MRL established value of 0.100 mg kg1 . These high
concentrations contribute to the average concentration
of 0.129 mg kg1 , which also exceeds the MRL.
Similar results were reported by Heukamp,16 who
found that coumaphos concentrations in the brood
chamber honey increased during the application to
even higher values, reaching 0.441 mg kg1 by the
end of the treatment. It must be mentioned that the
coumaphos concentration was below the MRL when
at each sampling time the frames were replaced with
new ones, so the residue results of the frames covered
the period since the preceding sampling day.16
In contrast, in the honey chamber samples, acaricide
residues were not detected until day 0 + 29, when
values up to 0.011 mg kg1 were measured, mainly

Pest Manag Sci (2007)

DOI: 10.1002/ps
E Karazafiris et al.
in samples from frames close to acaricide strips. On
the last day of treatment (0 + 42), residues reached
0.02 mg kg1 and remained below 0.02 mg kg1 until
the end of the trial. It is worth noting that Tremolada
et al.15 found 0.0006 mg kg1 of coumaphos residues
in honey 44 days after spraying with Perizin solution.
The difference in the results of the two studies could
be explained by the different formulation used in
each case, and by the fact that the amount of active
ingredient is released by CheckMite+ formulation
over a longer period.
The importance of collecting honey from the honey
super and not from the brood chamber is indicated by
this research. Since coumaphos is rather persistent in
honey, great care should be taken in the manipulation
of bees, especially when moving combs from one
chamber to another to relieve congestion or to apply
techniques like Demaree to control swarming.20
Concerning the efficacy of CheckMite+ strips
against the varroa mite, in a previous study the authors
found that, during the first 15 days of application,
91.4% of the mites were killed, while the percentage
of dead varroa reached 96.9% after 42 days.21 Average
residues in a single Langstroth hive were 0.090 and
0.129 mg kg1 at 14 and 42 days after treatment
respectively. According to the label instructions, strips
must remain in the colony for 42 days in order
to achieve maximum efficacy. Since the greatest
percentage (91.4%) of mites are killed during the first
14 days of application, it would be useful to examine
whether the residues in honey could be minimized by
reducing the days of treatment from 42 to 15.
Another concern with coumaphos residues, besides
the contamination of beehive products, should be
their impact on honeybee brood rearing. Colonies
treated with CheckMite+ contained coumaphos
residues averaging 45.5 mg kg1 in wax and
0.0030.008 mg kg1 in honey,22 and 6.1 and
5.9 mg kg1 of coumaphos in wax from brood
chambers and honey chambers of CheckMite+-
treated hives.16 The presence of coumaphos at high
concentration in wax causes queen failure. Collins
et al.23 found that queens could not develop in
beeswax cups containing 1000 mg kg1 of coumaphos,
and more than 50% of queen cells were rejected
at concentrations of 100 mg kg1 . Koeniger et al.24
also reported negative effects on queen rearing with
CheckMite+ strips.
The main goal of this research was to study
in vivo the behaviour and distribution of coumaphos
residues in the beehive, considering the loading of
honey production with coumaphos residues. The
authors did not examine residues in wax. Considerable
contamination of brood chamber honey was found,
especially of that derived from combs near the strips.
Considering published data on the behaviour of
coumaphos residues in beehive products, it is assumed
that a similar contamination of wax is likely. The
effects of coumaphos residues on queen rearing, and
the results from this study, raise concerns about the
impact of repeated application of CheckMite+ on the
development of all brood stages of honey bees.
To conclude, the results of the study show a
significant distribution of coumaphos residues in
honey within the various parts of a beehive. On the
last treatment day, residue levels in brood chamber
honey exceeded 0.100 mg kg1 only in samples from
combs next to CheckMite+ strips, and were lower in
honey obtained from the other combs. Also, a slight
contamination of the honey chamber was observed.
It is evident that higher residue concentrations are
found in the final product after acaricide application
as a slow-release formulation than as a spray solution.
Consequently, avoiding harvesting honey from combs
placed next to CheckMite+ strips and treating the
colonies for a shorter period will, provided efficacy
is not compromised, result in honey of acceptable
quality. Since the use of CheckMite+ strips is very
simple, beekeepers might be tempted to leave the
strips permanently in the hives. Such practices may
encourage development of mite resistance and the
presence of residues in honey.
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