Journal of Phytopathology and Pest Management 5(2): 1-11, 2018
pISSN:2356-8577 eISSN: 2356-6507
Journal homepage: http://ppmj.net/
∗
Corresponding author:
M.F. Abou-Ghadir,
E-mail: mfaboughadir@yahoo.com
1
Copyright © 2018
Evaluation of commercial honey bee
queens quality in Egypt
A.M. Ahmed
1
, A.M. Moustafa
1
, M.M. Khodairy
2
, M.F. Abou-Ghadir
2*
1
Plant Protection Research Institute, Agricultural Research Center, Dokki, Giza, Egypt
2
Plant Protection Department, Faculty of Agriculture, Assiut University, 71526 Assiut, Egypt
Abstract
Keywords: Apis mellifera, virgin queen, morphological characters, successful acceptance and mating process.
To evaluate numbers of morphological characteristics of virgin honey bee queens, a
total of 720 queens were obtained from five queen rearing stations were used. Queen
samples were collected in March and August during seasons of 2015 and 2016.
General characters as wet weight, thorax width and length, sum of third and fourth
tergites length, abdomen length, number of ovarioles and volume of spermatheca
were measured. The present result indicated that the queen weight was ranged from
134.33 to 156.34 mg in all tested samples. Insignificant difference of queen weights
between the two trials in March or August was recorded. The averages of ovariole
numbers were ranged from 118.93 to 130.11 in March, whereas they ranged from
125.32 and 131.26 in August. Both queen characters, weight and ovariole numbers
were under the international standard for queen quality. The general mean of
queen's acceptance percentage in August (83.33%) was differed significantly from
the acceptance percentage in March (69.35%). The percentage of successful natural
queen mating was ranged from 81.33 to 88.8 % in March and August, respectively.
The present results Manifest a lack of queen quality and queen rearing practices in
queen rearing stations in Egypt.
Ahmed et al., 2018
2
Introduction
Honey bee (
Apis mellifera
L.) is the most
beneficial insect in agriculture because it
produces honey and other valuable
products, and fulfills an important role as
a pollinator of crops. Approximately 35%
of arable crops depend directly on
pollinators (Klein et al., 2007). In recent
years much attention has focused on the
overwintering losses of the managed
honey bee population in the United States
and Europe (Pettis & Delaplane, 2010;
Potts et al., 2010; Van Engelsdorp &
Meixner, 2010) as well as in Egypt
(Moustafa et al., 2014; Hassan, 2009).
Because of the central role that queens
play within colonies, improving the
productivity and the health of honey bee
colonies is often synonymous with
improving the quality of queen. The
physical and internal characters of a
virgin honey bee queen are vital
components to the performance of the
colonies, which this queen is heading
later on. The present investigation is
aimed to evaluate the characteristics of
commercial honey bee virgin queens
obtained from different local rearing
sources and to examine the acceptance
percentage of them and the success of
natural mating as a first step for a
successful maintenance of honey bee
colony.
Materials and methods
The experimental works were conducted
in the laboratories of Agriculture
College, Assiut University, Assiut,
Egypt. Nuclei, equal in strength of
Carniolan honey bee hybrid (
Apis
mellifera carnica
) located in a private
apiary at Abnoub district, were used for
field studies during March and August,
2015 and 2016 seasons.
Virgin queen samples collection:
A
total of 720 virgin queens obtained from
five stations (Table 1); forty queens from
everyone at different local geographical
areas. The queen rearing stations were
coded by alphabetical letters. The queens
were gained on six-day from emergence
date as described by Tarpy et al. (2004).
Every queen was placed into standard
wooding cages and attended by few
honey bee workers until inspection.
Virgin queens from each source were
weighed individually using electrical
balance with 0.0001g accuracy. After
queen weighting, every sample was
divided into two groups: Group A: 25
queens for external and internal
morphological characters measurements,
Group
B: 15 queens for field studies
(acceptance and mating).
Table 1: The virgin queen sources and trial times of
introducing.
Sources
Cod
March
August
Assiut
(A)
+
+
Elgharbya
(B)
+
+
Menoufiya
(C)
+
+
Menoufiya
(D)
+
+
New Valley
(E)
+
-
External and internal body
characteristics:
Abdominal length,
maximal thorax width and length, and
the sum length of the third and fourth
tergite were measured. Every queen was
carefully dissected and numbers of
ovarioles of right ovary were counting
under stereoscopic self-illuminated
binocular according to the method of
Ibrahim (1977).
The diameter of
spermatheca was measured under
stereoscopic using a micrometer lens.
Spermatheca volume (SV) was
Ahmed et al., 2018
3
calculated according to the formula
(Hatch et al., 1999):
SV = (4/3) (π) (r
3
)
Where r = average radius of the r for the
length and the r for the width of the
spermatheca and π = 3.14.
Queen acceptance and natural mating:
The virgin queens obtained from
different source were introduced into
mating nuclei, contains five frames, using
a Benton mailing cage, mating nuclei
should be queenless two days before
introducing the virgin queens. The cage
was held between the frames with the
screen facing downward. Releasing of
the caged virgin queens was done after 2
days from the introduction. After 24
hours from releasing queens, the mating
nuclei were examined to determine the
queen acceptance percentage (Rhodes et
al., 2004). Accepted queens were
inspected daily until they started egg
laying, then, percentages of queen mating
success after queen releasing were
recorded (El-Sarrag & Nagi, 1989).
Statistical analysis:
Data were analyzed
using one-way analyses of variance by
MSTAT-C (1988) software package
while means were separated using the
least significant differences method,
when a significant "F" test was obtained.
The t-test was used to evaluate the
differences between the means, when
only two variables were tested.
Results
Virgin queens were obtained from
different sources in Egypt to assess
certain of their external and internal
characters in relation to the successful
acceptance in new nucleus. Queen
weight is often used as a good character
for overall the other variables of queen
quality. The average weight of the virgin
queens obtained from the five queen
rearing stations introduced in the
experimental nuclei at two trials on
March and August of both years 2015
and 2016 is presented in Table (2). The
recorded weights indicated numerical
and significant differences among the
inspected sources on the two trials.
Queens of A and E sources showed over
the heaviest weight in comparing with
other sources, while those of D source
presented the lowest weight. The general
mean of queen weight from A source on
March trial was 155.14 ± 14.70 mg over
other compared sources, while the lowest
134.33 ± 17.30 mg belonged to D source.
On August trial, the wet weight of queen
from A source gained 156.34 ± 14.11 mg
over the other compared sources, while
the lowest weight 145. 4 ± 16.32 mg
reported for the queens of D source. The
t-test supported the insignificant
difference of queen's weight introduced
on two trials on March and August
(Table 2).
Morphometrical characteristics of
virgin queens:
Data represented in Table
(3) show the variations in all
measurements of studied morphological
characteristics including the thorax
length and width, sum of third and fourth
tergites length and abdomen length
during March season of 2015 and 2016
years. The maximum length of thorax
was 4.93 ± 0.37 mm recorded for E
source, while the shortest length of 4.59
± 0.55 mm recorded for B source. The
Ahmed et al., 2018
4
maximum width of thorax was 4.11 ±
0.38 mm recorded for B source, while the
shortest length of 2.81 ± 1.42 mm
recorded for E source. The length of the
sum of third and fourth tergites was 3.68
± 0.50 mm recorded for A source, while
the shortest length was 3.18 ± 0.40 mm
and recorded for C source. The maximum
length of abdomen (8.71 ± 0.80 mm) was
recorded for E source, while the shortest
length (8.05 ± 0.54 mm) belongs to B
source. Data show significant differences
among the sources with respect to most
morphometric measurements of the
virgin queens at March introducing trial.
It can be concluded that all the longest
measurements were concerned to queens
of E source, while the lowest figures
described the queen of B source, during
March trial. Data represented in Table (3)
show also the results of August trial in
which the variations in all measurements
of studied characteristics including the
thorax length and width, the sum of third
and fourth tergits length, and the
abdomen length. The maximum length of
thorax was 4.98 ± 0.30 mm recorded for
A source, while the shortest length was
4.64 ± 0.26 mm which recorded for B
source. The maximum width of thorax
was 4.15 ± 0.25 mm and recorded for A
source, while the shortest length was
4.02 ± 0.26 mm and recorded for C
source. The length of the sum of third
and fourth tergites was 3.93 ±0.37 mm
and recorded for A source, while the
shortest length was 3.79 ± 0.44 mm and
recorded for C source. The maximum
length of abdomen was 8.95 ±0.44 mm
and recorded for A source, while the
shortest length (8.74 ±0.77 mm) was
recorded for E source. Data show
significant differences among the sources
with respect to most morphometric
measurements of the virgin queens in
August trail.
Table 2: Body weight of virgin queens obtained from different rearing sources, introduced into two trials March
and August of 2015 and 2016.
Production
Date
Mean of body weight (mg) ± SD
Virgin queen sources
Mean
±SD
A
B
C
D
E
March
146.81
±11.99 a
142.45
±18.96 a
144.95
±20.80 a
133.85
±17.48 a
145.99
±17.32a
142.81
±5.27 b
August
155.37
±14.53 a
136.66
±13.30 c
147.42
±26.18 ab
140.90
±21.72bc
-
145.09
±7.06 ab
March
163.47
±12.33 a
140.64
±14.74 c
135.28
±17.37 c
134.82
±17.35 c
155.39
±
11.80b
145.92
±12.86ab
August
157.31
±18.70 a
154.14
±14.41 a
150.14
±14.75 a
152.80
±16.11 a
-
153.60
±2.58 a
March
(2015&2016)
155.14
±14.70 A
141.56
±16.91 B
140.18
±19.66 B
134.33
±17.30C
150.62
±15.49A
144.34
±8.35 ns
August
(2015&2016)
156.34
±14.11 A
145. 4
±16.32 B
149.13
±20.90 B
146.60
±20.02 B
-
149.36
±4.74 ns
Means in the same row followed by different letters are significantly different, at 5% probability, ns:
not significant.
Ahmed et al., 2018
5
Table 3: Morphometric characteristics of virgin queens obtained from different rearing sources introduced in March and
August of 2015 and 2016.
Mean values (± SD) of external characters
Trials
Sources
Abdomen
Length
Tergits
(3
rd
+4
th
)
length
Thorax
Length
Width
March
2015&
2016
A
8.59±0.60 a
3.68±0.50 a
4.81±0.46 ab
3.91±0.40 a
B
8.11±0.61 b
3.26±0.43 c
4.59±0.55 c
4.08±0.38 a
C
8.05±0.54 b
3.18±0.40 c
4.72±0.29 bc
4.04±0.33 a
D
8.29±0.60 b
3.46±0.41 b
4.79±0.30 ab
4.11±0.30 a
E
8.71±0.80 a
3.67±0.35 a
4.93±0.37 a
2.81±1.42 b
August
2015&
2016
A
8.95±0.4 a
3.93±0.37 a
4.98±0.3 a
4.15±0.25 a
B
8.86±1.06 a
3.84±0.44 a
4.64±0.26 c
4.03±0.24 b
C
8.77±0.75 a
3.79±0.44 a
4.80±0.26 b
4.02±0.26 b
D
8.74±0.77 a
3.87±0.82 a
4.83±0.33 a
4.14±0.22 a
General mean
March
8.35±0.29 B
3.45±0.22 B
4.76 ± 0.12 ns
3.79 ± 0.55 ns
General Mean
August
8.83 ± 0.08A
3.85 ± 0.05 A
4.81 ± 0.12 ns
4.08±0.06 ns
Means in the same column followed by different letters are significantly different, at 5% probability, ns: not significant.
Internal characteristics of virgin
queens:
The virgin queens obtained from
different rearing sources were dissected
to count the ovariols. The average
numbers of ovarioles were presented in
Table (4). During the trial of March
(2015 & 2016), the highest number of
ovarioles (130.11 ± 12.60) was recorded
for E source, while the lowest number of
(121.32±12.26) ovarioles was recorded
for C source. When the trial was
impacted on August (2015 & 2016) the
queens of B source hold the highest
numbers of ovarioles over the other
compared sources with an average of
131.35 ± 13.65. The lowest average
(123.41 ± 10.02) was reported for queens
of C source.
Table 4: Ovarioles number of commercial virgin queens introduced in March and August 2015 and 2016 years.
Years
Production
Date
Ovarioles number /ovary (Mean ± SD)
Virgin queen sources
General mean
±SD
A
B
C
D
E
2015
March
120.81
±13.77 ab
114.00
±14.149 b
114.90
±11.360 b
118.47
±9.15 ab
123.4
±8.5a
118.32
±3.96 c
August
126.67
±9.02 ab
130.10±
14.05 a
121.63
±12.52 b
119.95
±9.88 b
-
124.59
±4.03 b
2016
March
130.05
±11.85 ab
128.86
±11.52 b
127.75
±14.52 b
133.19
±10.57 ab
137.1
±12.5a
131.39
±3.78 a
August
131.25±
11.21 ab
132.60
±13.41 a
125.20
±9.12 b
131.50
±11.76 ab
-
130.14
±2.89 a
General mean
March
(2015&2016)
125.43
±21.55 B
121.43
±22.06 B
121.32
±12.26 AB
125.83
±14.39 B
130.1
±12.6A
124.85
±4.35 ns
August
(2015&2016)
128.81
±10.04 AB
131.35
±13.65 A
123.41
±11.96 BC
125.72
±10.02 C
-
127.32
±3.06 ns
Means in the row followed by different letters are significantly different at 5% probability, ns: not significant.
Ahmed et al., 2018
6
Volume of the spermatheca:
The
average volume of the spermathecal, in
virgin queens from the rearing sources
introduced to nucleus through the months
of March and August, are presented in
Table (5). The highest volume of the
spermatheca volume in March trial (2015
& 2016) was (1.28±0.30 mm
3
) recorded
for E source while the lowest volume
(1.03±0.27 mm
3
) was recorded in B
source. On the other hand, the August
rearing supported that the highest
volume of the spermatheca (2015 &
2016) was (1.37±0.28 mm
3
) recorded for
A source while the lowest volume
(1.07±0.23 mm
3
) was recorded for B
source. The general mean of the
spermatheca volume in March was 1.14
mm
3
less numerically but not
significantly than 1.26 mm
3
of August
trial. Moreover, the E source did not put
in comparison during August.
Table 5: Spermatheca volumes (mm
3
) of virgin queens during two trials (March and August) of 2015 and 2016.
Years
Production
Date
Spermatheca volumes means (mm
3
) ± SD
Virgin queen sources
General mean
±SD
A
B
C
D
E
2015
March
1.15
±0.33 ab
0.94
±0.17 c
1.11
±0.18 b
1.14
±0.29 b
1.31
±0.35a
1.13
±0.13 a
August
1.43
±0.31 a
0.98
±0.22 b
1.29
±0.36 a
1.32
±0.31 a
-
1.25
±0.16 a
2016
March
1.25
±0.31 a
1.12
±0.32 b
1.11
±0.23 a
1.11
±0.28 a
1.25
±0.23a
1.16
±0.07 a
August
1.31
±0.25 ab
1.18
±0.21 b
1.24
±0.21a
1.35
±0.34 a
-
1.27
±0.06 a
General mean
March
(2015&2016)
1.20
±0.32 AB
1.03
±0.27 C
1.11
±0.20 BC
1.12
±0.28 BC
1.28
±0.30A
1.14
±0.09 ns
August
(2015&2016)
1.37
±0.28 A
1.07
±0.23 B
1.27
±0.31 A
1.33
±0.3 A
-
1.26
±0.11 ns
Means in the row followed by different letters are significantly different at 5% probability, ns: not significant.
Virgin queen’s acceptance:
The
present study was conducted to evaluate
the successful acceptance of virgin
queens obtained from different
commercial sources, through two trials in
March and August of 2015 and 2016
years. The obtained results are presented
in Table (6) and elucidate that the
general means of both March and August
of years 2015 and 2016 reported a high
respective percentages of 69.35% and
83.33% successful acceptance. A
fluctuated percentage of successful
acceptance was remarkable according to
the source of queens and the breeding
seasons within range between 46.6 to
100%. The general means of queen
acceptance percentage in August
(83.33%) was highly significant over the
mean of March trial (69.35%).
Virgin queens mating:
The successful
mating percentages of virgin queens
were calculated after laying eggs and
insuring in the established colonies. .
The obtained results (Table 7) indicated
that highly successful records for most
compared colonies that had successful
mating percentages ranged from 57.1 to
100%. The queens obtained from A, B
Ahmed et al., 2018
7
and E sources registered a full
percentage of successful mating, while
those from C and D got the lowest
position. The lowest successful mating of
57.1% was registered for D queens
introduced in August trial of 2015.
Moreover, the t-test supported the
insignificant difference between the
general means of the two trials in March
and August (Table 7).
Table 6: Percentages of queen's acceptance during two trials, March and August of 2015 and 2016.
Years
Production
Date
Virgin queens acceptance (%)
Sources
General mean (%)
A
B
C
D
E
2015
March
86.6
66.6
93.3
53.3
60.0
72.00
August
86.6
100
100
93.3
-
95.00
2016
March
80.0
46.6
53.3
80.0
73.3
66.70
August
73.3
73.3
73.3
66.6
-
71.66
General mean (%)
March (2015&2016)
83.3
56.6
73.3
66.6
66.6
69.35 ns
August (2015&2016)
80.0
86.6
86.6
80.0
-
83.33 ns
Table 7: Percentages of successful mating of virgin queens during two trials (March and August) of 2015 and 2016.
Years
Production
Date
Mating success (%)
Virgin queen sources
General mean (%)
A
B
C
D
E
2015
March
76.9
90.0
71.4
87.5
77.8
79.62
August
92.3
100.0
66.7
57.1
-
78.94
2016
March
100.0
100.0
87.5
100.0
100.0
98.00
August
90.9
81.8
72.7
90.0
-
83.72
General mean (%)
March (2015&2016)
88.4
95.0
79.3
93.75
88.9
88.80 ns
August
(2015&2016)
91.6
90.9
69.7
73.5
-
81.33 ns
ns: not significant.
Discussion
There are many problems currently
facing beekeeping in Egypt. One of these
problems is the overwintering losses of
the managed honey bee population. A
survey of beekeeping operations in
Assiut Governorate showed that
beekeepers classify the “poor queens” as
the second suspected cause for colony
losses (Moustafa, 2013). Similar survey
of certain characteristics of virgin honey
bee queens had been conducted by
Hamza (2015) but the current study is the
most comprehensive in the quantification
of physical characters of virgin queens
obtained from different rearing
commercial sources in relation to the
successful acceptance and mating of
queens. Lack of queen importance and
the current queen breeding practices may
reduced the genetic diversity of Egyptian
honey bee population. In turn, queen
breeders buy most of their queens from a
relatively small number of reliable
sources; import stock; and select within
their own gene pool. Weight of virgin
queen used by many investigators as a
good proxy among other morphological
parameters to evaluate their quality. In
this study queen weights found to be
ranged from 133.85 to 163.40 mg in
Ahmed et al., 2018
8
March rearing and from 136.66 to 137.31
mg in August rearing which is no
comparable with the data of Woyke
(1971), Nelson and Gary (1983), Van
Eaton (1986) and Delaney (2011).
Queen
weight is recommended criteria to assess
queen quality as it relates to different
management practices in commercial
queens
(Abd Al- Fattah et al., 2011;
Nelson, 1989; Kaftanoglu & Peng, 1980).
The weight of queens at emergence
showed a wide range of difference
because of factors such as age of larvae,
season, condition of rearing colonies and
racial differences (Skowronek et al.,
2004; Gençer et al., 2000; Weiss, 1974;
Woyke, 1971; Mirza et al., 1967).
Genetically different subfamilies within a
group of bee larvae can also contribute in
to the variability of emerged queens
(Moritz et al., 2005). There were
differences among the sources and
between the two rearing seasons in terms
of queen weight. The highest mean
weight was recorded when queens were
reared in late summer (August). Shawer
et al. (1980) found that the body weight
of virgin queens differed significantly
depending on the rearing season. They
also reported that queens produced in
Egypt during May and August were
heavier than those reared in other
months. Delaney et al. (2011) found the
mean wet weight for non-laying queens
to be 0.184±0.217g; they also reported
significant differences between the
various sources of queen bee suppliers.
Hegazy (1974) also mentioned that the
mean weight of queens was significantly
affected by rearing them in different
seasons. He recorded the maximum
weight in summer (July). Mustafa
et al.
(2002) indicated that the most suitable
seasons for rearing queens in Egypt were
late summer, followed by summer and
then spring. Data of the present study
showed that certain morphological
characteristics of queens were not
significantly affected by rearing them
either in March rearing or in August
rearing. The production and quality of
queens are affected by the rearing season
(Abd Al-Fattah et al., 2003 ; Hassan &
Mazeed, 2003; Abou El-Enain, 2000). It
was found that the numbers of ovarioles
in inspected queens were varied
significantly according to the source, but
only numerically among the introducing
time. The inspected queens introduced in
August trial had more ovarioles than
those of March trial. Data of Moustafa et
al. (2014) confirmed our results.
Moreover, the volume of spermatheca
was also varied in virgin queens from
different rearing sources, without
statistical difference, between the two
introducing trials. Hegazy (1974)
supported these results. The acceptance
rate of queens by queenless colonies is
very important for colony survival.
Beekeepers buy queens, and re-queen the
colonies in the spring or in the fall. If the
queens are not accepted, worker bees
raise new queens from existing larvae.
Lots of inexperienced beekeepers end up
losing their colonies in an effort to re-
queen them personally. There are many
reasons for rejection of queen
introduction. Moretto et al. (2004)
reported that there was some seasonal
variation in the acceptance rates.
Generally, there was greater acceptance
in months with good honey flows than
other months. Also, the acceptance of
introduced queens was influenced by
their origin. The rate of acceptance of
daughter queens from 11 different
mother queens varied significantly, and
Ahmed et al., 2018
9
ranging from 33 to 75%. There appears
to be a genetic influence of the mother
queen on the introduced queen's
acceptance rate. Gloria et al. (2007)
supported these results. The presence of
old or virgin queens in the colony will
always cause the rejection. Good quality
queens probably produce more
pheromones, start laying sooner and
become accepted
(
Rangel et al., 2016).
Moreover, the genetics of the colonies,
climatic conditions, nectar and pollen
flow, and queen introduction methods are
also important factors for the successful
queen introduction. The obtained results
indicated highly successful
records of
queens mating process for most
compared colonies, with numerical
variation among them.
Tarpy et al.
(2015) found that no significant
differences in mean mating frequency
between the feral and managed queens,
suggesting that queens in the remote,
low-density population of colonies in the
Arnot Forest are neither mate-limited nor
adapted to mate at an especially high
frequency. These findings support the
hypothesis that the hyper polyandry of
honey bees has been shaped on an
evolutionary timescale rather than on an
ecological one. These studies have shown
that queen reproductive capacity and
mating success are highly variable across
various commercial sources, because
these characteristics are impacted by
numerous factors that affect queen
development.
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