Assessing the Mating ‘Health’ of Commercial Honey Bee Queens

Author(s) :David R. Tarpy, Jennifer J. Keller, Joel R. Caren, and Deborah A.

Delaney
Source: Journal of Economic Entomology, 105(1):20-25. 2012.
Published By: Entomological Society of America
DOI:
URL: http://www.bioone.org/doi/full/10.1603/EC11276

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 APICULTURE AND SOCIAL INSECTS

Assessing the Mating ‘Health’ of Commercial Honey Bee Queens

DAVID R. TARPY,1,2 JENNIFER J. KELLER,1 JOEL R. CAREN,1 AND DEBORAH A. DELANEY3

J. Econ. Entomol. 105(1): 20Ð25 (2012); DOI: http://dx.doi.org/10.1603/EC11276

ABSTRACT Honey bee queens mate with multiple males, which increases the total genetic diversity
within colonies and has been shown to confer numerous beneÞts for colony health and productivity.
Recent surveys of beekeepers have suggested that Ôpoor queensÕ are a top management concern, thus
investigating the reproductive quality and mating success of commercially produced honey bee
queens is warranted. We purchased 80 commercially produced queens from large queen breeders in
California and measured them for their physical size (fresh weigh and thorax width), insemination
success (stored sperm counts and sperm viability), and mating number (determined by patriline
genotyping of worker offspring). We found that queens had an average of 4.37 ⫾ 1.446 million stored
sperm in their spermathecae with an average viability of 83.7 ⫾ 13.33%. We also found that the tested
queens had mated with a high number of drones (average effective paternity frequency: 17.0 ⫾ 8.98).
Queen “quality” signiÞcantly varied among commercial sources for physical characters but not for
mating characters. These Þndings suggest that it may be more effective to improve overall queen
reproductive potential by culling lower-quality queens rather than systematically altering current
queen production practices.

KEY WORDS honey bee, queen, insemination success, mating number

Honey bees (Apis mellifera L.) are one of the most
beneÞcial insects in agriculture because they serve as
invaluable pollinators of crops that are responsible for
an estimated 35% of the western human diet (Klein et
al. 2007, National Research Council 2007). In recent
years, much attention has focused on the overwinter-
ing lossesÑ both explained and unexplainedÑ of the
managed honey bee population in the United States
and Europe (e.g., vanEngelsdorp et al. 2009, Pettis and
Delaplane 2010, Potts et al. 2010, vanEngelsdorp
and Meixner 2010). Addressing overall colony health
and productivity is therefore of primary importance
for honey bee and apiculture research.
Because of the central role that queens play within
colonies, improving the productivity and health of
honey bee colonies is often synonymous with improv-
ing the quality of queens. Indeed, many standard tech-
niques of bee management focus on maximizing the
reproductive potential of queens. For example,
Ôswarm box cell-starterÕ colonies (Laidlaw and Page
1997), used to initiate queen rearing in commercial
operations, maximizes the number of nurse bees that
produce royal jelly to feed developing gyne brood.
However, most advances in improving queen quality
have been accomplished by various bee breeding pro-
grams, which typically use the instrumental insemi-
nation technique to artiÞcially select certain colony
1 Department of Entomology, Campus Box 7613, North Carolina
State University, Raleigh, NC 27695-7613.
2 Corresponding author, e-mail: david_tarpy@ncsu.edu.
3 Department of Entomology and Wildlife Biology, University of
Delaware, 252 Townsend Hall, Newark, DE 19716.
phenotypes. Numerous strains of honey bees have
been developed, each exhibiting an assortment of se-
lectable traits, such as disease- or mite-tolerance, gen-
tleness, honey production, and hygienic behavior
(e.g., Harbo and Harris 1999, Rinderer et al. 2001,
Spivak and Reuter 2001). The goal of these programs
has been to improve queen genetic quality, which, in
turn, improves overall colony productivity.
Independent of genetic stock, a colony can be af-
fected by numerous factors that greatly diminish the
reproductive potential of its queen. First, a queen can
be replaced by her worker offspring during superse-
dure events, typically when an aging queen depletes
her sperm supply (Butler 1957). Supersedure queens
are of lower quality compared with those raised under
more favorable conditions. For example, Kostarelou-
Damianidou et al. (1995) showed that colonies headed
by replacement queens produced up to 34% less brood
and 36% less honey. To maximize queen and colony
quality, the standard advice to beekeepers is to man-
ually replace their queens every spring. Such proce-
dures can be expensive both directly (for the costs of
the queens themselves) and indirectly (accidental
queen and colony loss, increased labor, and dimin-
ished honey yields), thus minimizing how often col-
onies need to be restocked will beneÞt beekeepers.
Second, a queen may produce brood that is not highly
viable. This is evidenced by “shotgun brood patterns,”
which are routinely observed by beekeepers and re-
sult in a colonyÕs inability to rapidly increase their
worker force and thus decreases productivity (Woyke
1980, Tarpy and Page 2002), usually as a result of local

0022-0493/12/0020Ð0025$04.00/0 䉷 2012 Entomological Society of America

February 2012 TARPY ET AL.: MATING HEALTH OF QUEEN BEES
inbreeding or low population-wide genetic diversity.
Therefore, increasing the health and quality of a queenÕs
brood has direct beneÞts for beekeepers. Finally, a col-
ony can host a litany of parasites and pathogens that
signiÞcantly diminish its productivity and overall health.
Like most domesticated animals, honey bees may ac-
quire any number of infectious agents as a consequence
of spatial overcrowding and equipment sharing. The cost
to prevent and treat disease is considerable, and a central
focus of apicultural research has been to reduce the
impact of economically important pests. Such ap-
proaches have relied heavily on chemotherapies, which
can have negative effects on queens (Haarmann et al.
2002) and hive products (e.g., Tsigouri and Menkissoglu
2001). Moreover, there is mounting evidence that many
pests are becoming increasingly resistant to the more
common treatments (Colin et al. 1997, Miyagi et al. 2000,
Mozes et al. 2000), making them less reliable and, ulti-
mately, ineffective.
Each of these concerns may be signiÞcantly impacted
by a single factor: how well a queen has mated. Multiple
mating by queen honey bees has been shown to 1)
mitigate the negative impacts of poor brood patterns as
a result of homozygosity at the sex locus and inbreeding
(Tarpy and Page 2002); 2) increase the number of stored
sperm (Schluns et al. 2005, Delaney et al. 2011), as well
as enhance queen attractiveness and pheromone proÞles
(Richard et al. 2007), thus prolonging longevity and re-
ducing the likelihood of supersedure; and 3) delay the
spread of infectious diseases among nestmates (Palmer
and Oldroyd 2003, Tarpy 2003, Seeley and Tarpy 2007).
As such, increasing the “mating health” of honey bee
queens will likely beneÞt beekeepers by simultaneously
increasing queen longevity, brood viability, and disease
tolerance.
A niche market within the apiculture industry is the
mass production and sale of open-mated queens to
beekeepers. Commercial queens typically sell for $15
to $25 each, and upwards of one million queens are
raised and sold every year in the United States (Cobey
et al. 2011). However, there have been surprisingly
few studies that have systematically and thoroughly
assessed mating frequency, insemination success, and
morphometric characters of queens produced by the
commercial queen-production industry in the United
States. Camazine et al. (1998) investigated the health
of commercially purchased queens in the United
States. In addition to bioassays of two adult parasites,
they estimated the stored sperm number of over 300
queens from many different sources. In that sample,
almost one in Þve queens were “poorly mated,” de-
Þned as fewer than three million stored sperm
(Woyke 1962). While very telling, these Þndings pro-
vide a limited and somewhat outdated view of the
mating health of commercial honey bee queens.
The physical and reproductive health of the queen
honey bee is a vital component to a colonyÕs well-
being, both genetically and socially. Commercial
queen breeders supply most of the queens used by
migratory beekeepers who move thousands of honey
bee colonies along pollination routes across the coun-
try. The commercial and hobbyist beekeepers alike
21
face many problems that challenge the health and
longevity of their stocks. Annual colony losses, in-
creases in chronic infections, and poor queen perfor-
mance have caused the apiculture industry to question
the quality of commercially produced queen honey
bees. Several surveys of U.S. beekeepers (vanEngels-
dorp et al. 2008, 2010, 2011) have consistently identi-
Þed “poor queens” as one of the top management
concerns in the apiculture community. While symp-
tomatic of many management problems, premature
supersedure and stored-sperm depletion are typical
complaints attributed to poor queens. Delaney et al.
(2011) recently found that commercial queens sam-
pled from Western and Southeastern queen breeders
were sufÞciently inseminated (⬇4 million sperm)
with a high average number of drones (⬇16 mates),
but they only tested 24 queens (two from each sam-
pled source) for their mating numbers. In this current
study, we concurrently examine the physical, insem-
ination, and mating quality of a large sample of com-
mercially produced queens.
Materials and Methods
Sampling. We purchased 10 queens each from
seven independent commercial queen producers lo-
cated in CaliforniaÕs central valley. All sources were
generically coded to remain blind and retain anonym-
ity. The genetic stock of all purchases was standard
ÔItalianÕ (presumably A. m. ligustica L.) honey bees,
although we purchased an additional 10 ÔCarniolanÕ
queens (presumably A. m. carnica) from Source C,
resulting in a total of 80 experimental queens. The
commercial operations were not informed as to
the purchasers (North Carolina State University
[NCSU]) or the beesÕ use in a scientiÞc study (all
queens were ordered by an individual in our program
and shipped to his private residence). Therefore, this
represents a random sampling of queens that are com-
mercially available from each operation.
Upon arrival at NCSU, we introduced each queen
into her own Þve-frame nucleus colony at the Lake
Wheeler Honey Bee Research Facility following stan-
dard methods. After their acceptance by the workers,
we permitted each queen to lay eggs for 3Ð5 wk before
sampling all queens within a source group. From each
colony, we caged a frame of capped brood and
emerged them in an incubator set at 34⬚C and ⬇50%
RH to sample newly emerged worker daughters from
each queen, which were subsequently frozen at
⫺80⬚C for paternity analysis (see below).
Morphological Measures. We measured all queens
from a given commercial source concurrently once
their brood was sampled (see Delaney et al. 2011). We
immobilized each laying queen by temporarily placing
her in a freezer and then recorded her fresh weight to
the nearest 0.1 mg on a digital scale. We then measured
her thorax width and head width to within 0.1 mm
using digital calipers, then sacriÞced each queen by
decapitation, dissected the queenÕs abdomen, and re-
moved her spermatheca. We measured the diameter
of the spermatheca using a calibrated ocular in a dis-
22 JOURNAL OF ECONOMIC ENTOMOLOGY
Table 1. Comparisons of morphological characters of commercial queens
Fresh wt.
Source n
(mg)
A 10 234.4 ⫾ 16.30a
B 8 221.0 ⫾ 15.27a
C(C) 7 233.0 ⫾ 15.42a
C(I) 6 225.8 ⫾ 14.96a
D 7 220.1 ⫾ 16.37ab
E 7 221.2 ⫾ 20.58a
F 9 195.9 ⫾ 11.42bc
G 5 191.1 ⫾ 7.90c
Avg. 218.7 ⫾ 20.66
Different commercial sources are coded for aninimity, both Italian (I), and Carniolan (C) queens were purchased from source C and were
analyzed separately. Different letters after each measure signify statistically signiÞcant differences using Tukey post hoc tests at ␣ ⫽ 0.05.
secting microscope and calculated its spherical vol-
ume (see Hatch et al. 1999).
Sperm Counts and Viability. To quantify stored
sperm number and viability, we followed the staining
protocol of Collins and Donoghue (1999). Immediately
after dissection, each spermatheca was ruptured in 0.5 ml
1.0 M HEPES. The ruptured spermatheca was removed
and the remaining sperm and HEPES was transferred
into a 2 ml vial. We added 10 ␮l of Sybr 14 in DMSO to
the vial and incubated the vial for 7 min in a 36⬚C water
bath. We then added 5 ␮l of propidium iodide and again
incubated the vial for 7 min in a 36⬚C water bath
(adapted from LIVE/DEAD Sperm Viability Kit, Mo-
lecular Probes, Eugene, OR). The stained sperm (⬇10 ␮l
of solution) was immediately loaded onto a hemacytom-
eter. A Zeiss Axioskopepi ßuorescent microscope (Carl
Zeiss, Hanover, MD) with a digital camera and FITC and
Rhodamine Þlters was used to visualize both live and
dead sperm. Live sperm ßuoresce at green wavelengths
(Em 520Ð570 nm) while dead sperm ßuoresce at red
wavelengths (Em 650 nm). We took Þve separate pic-
tures (nonoverlapping Þelds of view) using Þrst the
FITC Þlter and then the Rhodamine Þlter. The numbers
of live and dead sperm were counted for each picture
under the differing Þlters and the percent of viable sperm
present from each spermatheca was calculated. The av-
erage sperm count from the Þve Þelds of view was cor-
rected for dilution to calculate the total number of stored
sperm.
Mating Numbers. We calculated the observed mat-
ing number and effective paternity frequency for each
queen following Tarpy et al. (2010). Brießy, we ex-
tracted whole genomic DNA from the legs of up to 48
workers from each colony using Chelex 100 resin
(Walsh et al. 1991) and subjected it to two multiplexed
polymerase chain reactions (PCRs) for eight microsat-
ellite loci (Am010, Am043, Am052, Am059, Am061,
Am098, Am125, and Am553). We analyzed the PCR
products on a 3730 ABI sequencer through the NCSU
Genomic Science Laboratory, and we inferred the pa-
ternity of each worker using COLONY (Wang 2004)
from which we were able to calculate the observed
mating number (No) and the effective paternity fre-
quency (me) of each queen following Nielsen et al.
(2003).
Statistical Analyses. We compared the different
commercial sources using one-way analysis of vari-
Vol. 105, no. 1
Thorax width Head width Spermatheca
(mm) (mm) vol (mm3)
4.43 ⫾ 0.295a 3.60 ⫾ 0.269a 0.90 ⫾ 0.142bc
4.46 ⫾ 0.265a 3.59 ⫾ 0.232a 1.09 ⫾ 0.181ab
4.41 ⫾ 0.188ab 3.51 ⫾ 0.182ab 1.13 ⫾ 0.114a
4.45 ⫾ 0.067ab 3.44 ⫾ 0.190ab 0.93 ⫾ 0.050abc
4.36 ⫾ 0.221ab 3.38 ⫾ 0.211ab 0.90 ⫾ 0.176bc
4.24 ⫾ 0.192ab 3.46 ⫾ 0.238ab 0.95 ⫾ 0.123ab
4.13 ⫾ 0.111b 3.24 ⫾ 0.161b 0.73 ⫾ 0.079c
4.26 ⫾ 0.069ab 3.37 ⫾ 0.178ab 0.88 ⫾ 0.134abc
4.34 ⫾ 0.226 3.45 ⫾ 0.233 0.94 ⫾ 0.175
ance (ANOVA) with Tukey post hoc tests. We then
used multivariate analyses using pairwise correlations
to quantify the relationships among all measures of
queen quality. We then conducted a Principal Com-
ponent Analysis on the covariances of the measure-
ments and saved the top two PCs for comparisons
among the different commercial sources. All means
are reported with ⫾1 SD unless noted otherwise, and
all statistical comparisons are two-tailed with a signif-
icance of ␣ ⫽ 0.05.
Results
Several of the queens did not survive original ship-
ment, and several more were either not accepted by
the workers in their introduced nucleus colonies or
died before sampling, thus a total of 61 queens were
analyzed. There was no signiÞcant pattern of queen
mortality with respect to their purchased source and
thus were common losses experienced with overnight
shipping of queens. There were signiÞcant differences
among the sources with respect to all morphological
measurements of the queens, including fresh weight
(F ⫽ 7.74; df ⫽ 7, 51; P ⬍ 0.0001), thorax width (F ⫽
2.79; df ⫽ 7, 51; P ⬍ 0.05), head width (F ⫽ 2.64; df ⫽
7, 48; P ⬍ 0.05), and spermatheca volume (F ⫽ 7.19;
df ⫽ 7, 51; P ⬍ 0.0001; Table 1).
When comparing measurements of queen insemi-
nation, however, there were less pronounced differ-
ences among queen producers (Table 2). Overall, the
average number of stored sperm in the 59 queens
examined was 4.37 ⫾ 1.446 million, and there were no
signiÞcant differences in sperm counts among the
sources (F ⫽ 1.18; df ⫽ 7, 51; P ⫽ 0.33). While one
queen from Source E was shown to still be a virgin (no
detectable stored sperm), only eight queens (13.6%)
were Ôpoorly inseminatedÕ (i.e., had fewer than three
million stored sperm). Given the calculated volume of
the spermatheca (see above) and the average size of
an individual spermatozoon, we also calculated the
percentage of the maximum Þlled capacity of each
queen following (Tarpy et al. 2011). The average for
all queens was 47.0 ⫾ 20.05%, and there were no
signiÞcant differences among queen sources (F ⫽ 1.39;
df ⫽ 7, 51; P ⫽ 0.23). There were, however, highly
signiÞcant differences in the percentage of viable
sperm across sources (F ⫽ 7.24; df ⫽ 7, 49; P ⬍ 0.0001),
February 2012 TARPY ET AL.: MATING HEALTH OF QUEEN BEES 23

Table 2. Comparisons of insemination and mating characters of commercial queens

Stored sperm Sperm Percent Observed Effective paternity

Source n
(⫻106) viability (%) Þlled mating no. (No) frequency (me)
A 9 4.96 ⫾ 1.648 92.6 ⫾ 4.60ab 54.1 ⫾ 28.26 20.3 ⫾ 5.20 21.4 ⫾ 9.40
B 8 4.37 ⫾ 1.582 78.0 ⫾ 7.44b 39.0 ⫾ 12.79 18.1 ⫾ 4.94 16.4 ⫾ 7.94
C(C) 7 4.98 ⫾ 1.113 85.4 ⫾ 8.24ab 42.6 ⫾ 9.94 20.2 ⫾ 5.42 21.8 ⫾ 11.77
C(I) 6 3.54 ⫾ 1.452 86.0 ⫾ 8.86ab 36.4 ⫾ 14.39 16.7 ⫾ 5.32 14.6 ⫾ 9.20
D 7 4.43 ⫾ 1.018 82.8 ⫾ 9.38ab 49.9 ⫾ 18.02 16.3 ⫾ 2.87 11.4 ⫾ 6.32
E 7 3.67 ⫾ 1.748 79.6 ⫾ 5.96ab 38.4 ⫾ 18.22 20.0 ⫾ 4.84 21.0 ⫾ 8.58
F 9 3.95 ⫾ 1.005 94.0 ⫾ 1.73a 52.8 ⫾ 14.06 15.8 ⫾ 4.66 12.2 ⫾ 5.24
G 6 4.96 ⫾ 1.710 59.4 ⫾ 26.47c 60.5 ⫾ 31.34 17.6 ⫾ 4.04 15.1 ⫾ 8.59
Avg. 4.37 ⫾ 1.446 83.7 ⫾ 13.33 47.0 ⫾ 20.05 18.2 ⫾ 4.84 17.0 ⫾ 8.98

Coded sources are the same as in Table 1. Different letters after each measure signify statistically signiÞcant differences using Tukey post
hoc tests at ␣ ⫽ 0.05, which were only signiÞcant for sperm viability and none of the other variables.

where the average sperm viability was 83.7 ⫾ 13.33 but
with some queens having as low as 20.5% viable sperm.
There was a signiÞcant effect of processing date on
sperm viability (F ⫽ 4.09; df ⫽ 5, 51; P ⬍ 0.005), but
there was no consistent downward trend over time so
that temporal effects on sperm longevity (e.g., Burley
et al. 2008) was likely not a factor.
We genotyped a total of 2,798 individual workers
(45.9 ⫾ 1.97 per colony) and assigned them to their
respective patriline within their colonies to infer
queen mating numbers. Overall, the mating numbers
of the tested queens were higher than expected (Ta-
ble 2). The average observed mating number was
18.2 ⫾ 4.84 and the average effective paternity fre-
quency was 17.0 ⫾ 8.98, both of which are numerically
(but not statistically) higher than their global averages
for the species (Tarpy and Nielsen 2002). As with the
insemination measures, there were no signiÞcant dif-
ferences in mating frequency across commercial
sources (observed: F ⫽ 1.22; df ⫽ 7, 53; P ⫽ 0.31;
effective: F ⫽ 1.89; df ⫽ 7, 53; P ⫽ 0.09).
There were many statistically signiÞcant correla-
tions among the measured variables, particularly
among the morphological measurements (data not
shown). Similar to Schluns et al. (2005) and Delaney
et al. (2011), we found a signiÞcant nonlinear (loga-
rithmic) relationship between effective paternity fre-
quency and stored sperm number (F ⫽ 7.97; df ⫽ 1, 55;
P ⬍ 0.01). We did not, however, verify the observed
relationships between thorax width and sperm num-
ber (F ⫽ 2.97; df ⫽ 1, 57; P ⫽ 0.09) or mating frequency
(F ⫽ 1.03; df ⫽ 1, 56; P ⫽ 0.31) that were reported in
our previous study (Delaney et al. 2011).
No singular variable can adequately capture queen
reproductive potential in its entirety, and thus many
different measurements can each reßect a different
facet of this complex quantitative trait. A proxy of
these measures can be generated in multi-dimensional
space using Principle Component Analysis (PCA), in
essence to collapse multiple correlated variables into
one or a few composite variables. PCA on the cova-
riances of the nine measured variables yielded two
prominent principal components that accounted for
99.2% of the total variance in queen reproductive
potential, the Þrst (PC1) accounting for 81.6% of the
total variance and the second (PC2) accounting for an
additional 17.6%. The loading variables (eigenvectors)
for PC1 were all positive (except for percentage
Þlled), whereas PC2 was negatively associated with all
morphometric characters (listed in Table 1) and pos-
itively associated with all “mating” variables (listed in
Table 2). Overall, there was no signiÞcant correlation
between the two principal components (r2 ⫽ 0.049;
P ⫽ 0.66), although there was one commercial source
where there was a strong negative relationship
(Source G: r2 ⫽ ⫺0.807; P ⬍ 0.005). A one-way
ANOVA on PC1 demonstrates signiÞcant differences
among queen sources for this single composite of
queen reproductive quality (F ⫽ 7.28, df ⫽ 7, 46, P ⬍
0.0001; Fig. 1), but there were no differences for PC2
(F ⫽ 1.07; df ⫽ 7, 46; P ⫽ 0.40).
Discussion
The intent of this study was to determine if commer-
cially produced honey bee queens are of low reproduc-
tive potential and may therefore explain their diminished
quality perceived by the industry. However, our data
shows very little, if any, evidence that this is the case for
newly mated queens. These Þndings corroborate Dela-
ney et al. (2011), who also suggest that commercially
Fig. 1. Plot of the two principal components (PC1 and
PC2) grouped by commercial source. No one variable ade-
quately measures queen reproductive potential, but a compos-
ite of many different measures can be generated in multi-di-
mensional space using PCA. PCs were generated from a
principal component analysis on the covariances of all nine
measured variables (Tables 1 and 2) and together explain over
99% of the overall variation in queen quality. Ellipses signify 50th
percentiles on both axes for each identiÞed source.
24 JOURNAL OF ECONOMIC ENTOMOLOGY
produced queens are generally high in their reproduc-
tive potential. The current population of queens had very
low levels of the gut microsporidian Nosema, as only
three of the 59 queens tested had detectable levels as
determined by PCR analysis following (Webster et al.
2008, data not shown). However, because the queens
were sampled 3Ð5 wk after being in nucleus colonies, and
we did not sample for Nosema among the workers in
those colonies, we cannot determine if they were in-
fected prior or subsequent to colony introduction. Nev-
ertheless, it is quite clear that contemporary commercial
queens have signiÞcantly lower parasite loads than in the
past (see Farrar 1947, Furgala 1962, Liu et al. 1987, Ca-
mazine et al. 1998).
We did not consistently observe strong correlations
between body size and mating success. Delaney et al.
(2011) found signiÞcant relationships between thorax
width and both stored sperm and effective paternity
frequency, but fresh weight did not show a similar
correlation (see also Tarpy et al. 2011). In the current
study, we found a marginally signiÞcant relationship
between weight and mating number (No: r2 ⫽ 0.260,
P ⬍ 0.05; me: r2 ⫽ 0.245, P ⫽ 0.06), but no such
relationships with thorax width. It is possible that
sample size may explain these differences; the current
study compared a total of 61 queens for all morpho-
logical and mating variables, whereas Delaney et al.
(2011) tested a total of 117 queens but only 22 of them
for mating number. Thus the current data set is more
robust for mating number but less robust for size
variables and sperm number. In any event, it seems
that queen size does have signiÞcant albeit weak re-
lationship to mating success (Tarpy et al. 2011).
The potential economic implications of our current
Þndings are fairly clear. There is more variation in queen
reproductive potential within operations than there is
among them; thus, efforts aimed at improving overall
queen quality would be better suited at culling low-
quality queens within an operation rather than imple-
menting a systematic management change. In other
words, if the goal of the queen-rearing industry is to
increase the average quality of queens, then it would
likely be more effective to remove the low end of the
distribution rather than shift it upwards. This places a
premium on developing technologies that can rapidly
assess queen quality (particularly mating success) with-
out destructive sampling (e.g., Webster et al. 2009) so
that subsamples of queen cohorts can be vetted before
their commercial sale. This would provide substantial
economic opportunities for increased prices of graded
queens, which would beneÞt breeders and beekeepers
alike. Such an effort would beneÞt from consolidated and
experimentally induced measures of queen quality so as
to rate all commercial queens on a single- or two-dimen-
sional measurement scale (Tarpy et al. 2011).
Our large-scale study indicates that the ongoing issues
of poor queens experienced in the industry (e.g., vanEn-
gelsdorp et al. 2008) is unlikely because newly mated,
commercially produced queens are systemically of low
reproductive potential. Thus, we do not currently have
evidence as to why the perception among beekeepers is
that overall queen quality has diminished in recent years.
Vol. 105, no. 1
Therefore, it is likely that if queen quality is an important
factor in honey bee management, then testing the nu-
merous environmental factors that can inßuence queen
productivity after mating should take priority. Future
research should emphasize the potential roles of factors
that affect queen quality before (e.g., genetic bottle-
necks, pedigree relationships among breeding popula-
tions) and subsequent to mating (e.g., mechanisms of
supersedure, capacity to store live sperm) to address
these concerns.
Acknowledgments
We thank John Harman, Winnie Lee, Flora Lee, Mithun
Patel, and Matt Mayer for their help in DNA extractions and
PCR analyses. We also express our appreciation and gratitude
to Marla Spivak, the CA Bee Breeders, and the NC State
Social Insect Working Group for their cooperation and many
helpful discussions subsequent to the collection of queens
used in this study. Special thanks goes to Laura Mathies for
use of her ßuorescent microscope for the sperm analyses
without which those measures would have been impossible.
This study was supported by the North Carolina Department
of Agriculture and Consumer Services and grant number
2007-02281 from the National Research Initiative of the
USDA Cooperative State Research, Education and Extension
Service awarded to DRT.
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Received 17 August 2011; accepted 6 October 2011.