J. Asia-Pacific Entomol.

8(3): 309-317 (2005)

www.entomology.or.kr
INDUSTRIAL ENTOMOLOGY
Mass Rearing of Apis cerana F. Queen
Dharam Pal Abrol*, R. M. Bhagat and Devinder Sharma
Division of Entomology, Faculty of Agriculture, Sher-e-Kashmir University of Agricultural Sciences and Technology,
Udheywalla, Jammu 180 002, Jammu & Kashmir, India
Abstract The conditions that determine the success of
mass rearing of Apis cerana F. queens were studied.
It was found that artificial queen cell cups with the
internal diameter of 6.2mm at base 8.6mm at the mouth
and 8.8mm depth were highly preferred by the bees
for rearing of queens from the grafted larvae. Likewise,
the wax obtained from old comb foundation was pre-
ferred over fresh comb foundation. Maximum accep-
tance was recorded for 12 and 6 number oflarval grafts.
High percentage and mean volume of queen cells. w ~ s
obtained from 12-6hr. old grafts. However, no sigru-
ficant differences were observed between grafts and
those provided with royal jelly. The same was true for
single and double grafts. The percentage acceptance
was in the order: March, April, August, and September.
Key words Apis cerana, cell cup materials, effect of
larval age, mass rearing, number of grafts, queens,
seasonal variations
Introduction
Productivity of a honey bee colony depends upon
the quality ofthe queen ofthe colony (Laidlaw, 1979;
Morse, 1979; Ruttner, 1986). Therefore, bee stocks
must be headed by the queen - having high fecundity
and oviposition pattern, gentle temperament, high
industriousness, longevity and disease resistance qua-
lities. Besides, queens may be required in large num-
ber for breeding and marketing purposes. Though
mass rearing of queens has extensively been studied
but mostly focused on Apis mellifera (Laidlaw and
Eckert, 1962; Johansson and Johansson, 1978: Gary
1979; Laidlaw, 1979; Morse, 1979; Ebadi and Gary,
1980; Kither and Pickard, 1983; Ruttner, 1986). Very
little is known about this technique in Apis cerana.
(Bhat, 1983; Wongsiri et al., 1988; Wongsiri and
Pthichot, 1990; Verma and Sharma, 1997). The pre-
sent paper reports the mass rearing of A. cerana.
Corresponding author.
E-mail: cispa@kangwon.ac.kr
Tel: +91-191-2462451; Fax: +91-191-2462982
(Received August 11, 2005; Accepted September 10, 2005)
Materials and Methods
The studies were conducted at Research Sub-Station
of Sher-e-Kashmir University of Agricultural Sciences
& technology located at Bhaderwah about 210 km
from Jammu city. Nine colonies of Apis cerana were
selected for the study. All the colonies were of 7-8
frames strength with sufficient stores of honey and
pollen. For queen rearing, the method of Laidlaw
(1979) was followed. Each set of experiment con-
sisted of three replications with one replication per
colony. The effect of following parameters on the
acceptance of queen cells was studied.
Size of artificial queen cups
Preparation of artificial Queen cell cups. Twenty
empty queen cells build under natural conditions
from honey bee colonies were collected during swar-
ming seasons (March). The dimensions of these na-
turally built queen cells was taken as a base and
three different sized cell forming rods were made
from seasoned hard wood. Bee wax used for making
artificial queen cell cups was obtained from old ho-
ney bee combs. Wax was wrapped in a muslin cloth
and immersed into water contained in a beaker, in
a water bath at 63-64C. The molten wax peculated
through the muslin into the beaker which was then
put into a trough of cold water. A thick layer of wax
which got solidified at the surface of the water was
removed. This wax was then placed in a 100 ml
beaker and was melted through indirect heat method
by placing the beaker over the water bath maintained
at 65C, just above the melting point.
The queen cell cups were prepared with the help
of cell forming rods. The rod of desired size was
first immersed into the cold water and after draining
excess water the rod was dipped up to desired depth
into the molten wax. The rod was then lifted and
the wax layer was allowed to set. The rod containing
wax layer was dipped again in the molten wax but
this time to a slightly lesser depth then the first dip.
310 1. Asia-Pacific Entomol. Vol. 8 (2005)
The process was repeated five times and every time
the depth to which the rod was dipped was reduced.
After the final dip, the rod was immersed into cold
water and the cell cup so formed was then removed
by twisting the rod.
Effect ofcell cup size. In order to determine the effect
of size of cells accepted, three types of cups were
prepared (Table 1).
Collection of royal jelly
A strong bee colony with sufficient young brood, nurse
bees and pollen stores was dequeened and allowed
to raise queen cells. Before sealing, the queen cells
were cut and larvae from the cells were removed.
A little warm distilled water was added to each such
cell and royal jelly was stirred. This diluted royal
jelly was taken out by means of a dropper and col-
lection in a small vial which was kept in the freezer
for further use. At the time 0: grafting a small drop
of this diluted royal jelly was placed at the bottom
of the cell cup with the help of a dropper to prime
the artificial queen cell cups R1d immediately a larva
was grafted into such a cell cup over the drop of
royal jelly.
Collection of larvae of desired age
The worker cells with freshly laid eggs were marked
with the marking fluid to obtain eggs of known age.
Marking fluid was prepared by dissolving dyes in
Table 1. Dimensions of queen cell cups
thick solution of shellac in absolute ethanol. This fluid
was applied to the margins of the cells with the help
of a fine camel hair brush. Marked egg combs were
then introduced into another colony (incubator colony)
for the development of the larvae. A bee colony with
sufficient nurse bees, pollen, sealed and emerging
brood was chosen for this purpose. The young brood
in this colony, if any, was removed. The incubator
colony was regularly provided with 40 % sugar syrup.
The pollen supplement was given at the time of pollen
dearth. After having removed the first batch of eggs
from a breeder colony another empty comb was in-
serted in its place and this sequence was repeated
to get a regular supply of eggs of known age.
Grafting of larvae
Honey bee colony with sufficient nurse bees, sealed
and emerging brood, and sufficient pollen and honey
stores was selected as a cell builder colony for rearing
queen honey bees from grafted larvae. The colony
was dequeened and young brood, if any, was removed.
Such a colony was liberally provided with 40 % sugar
syrup throughout the course of experiment starting
from one day prior to the introduction of grafted
larvae. Desired numbers of wooden bars were fitted
in the standard deep frame (interior length 26.5 em
and interior breadth 18 em) for building the queen
cell cups. Required number of wax blocks 0.5 em
thick, 1 em wide and 1.5 em long were fixed on the
lower surface of these bars to serve as bases for queen
cell cups. The queen cell cup bases were dipped into
the molten wax and were fixed over these blocks.
These frames containing cell cups were then intro-
Queen cell cup Diameter at base(mm) Internal diameter at mouth(mm) Depth (mm)
A 5.4 7.8 7.8
B 7.7 8.7 15.2
C 6.2 8.6 8.8
Table 2. Acceptance of larvae grafted in queen cell cups of different dimensions for rearing A. cerana queens
Number of grafts on Total
Size Top Bar Central Bar Lower Bar
A NA PA A NA PA A NA PA Graft A NA PA
A 0 12 0 0 12 0 0 12 0 36 0 36 0
B 4 8 33.33 4 8 33.33 5 7 41.7 36 13 23 36
C 7 5 58.33 9 3 75.00 8 4 66.70 36 24 12 66.66
Mean 3.67 8.33 30.55 4.33 7.67 36.11 4.33 7.67 36.11 36 12.33 23.66 34.26
A= Accepted; NA ~ Not accepted; PA =, Percent accepted, X'(cal) =35.60; X'(2 df, 0.05) ~ 5 . 9 9 0 ;
duced into the cell building colony for polishing and
smoothening of the queen cell cups at least for 24
hours before larvae were grafted into them. In the
frame holding cell cups the top most bar was fitted
at a distance of 5 em from the upper edge of the
frame and the second bar was fixed at a distance of
7 em from the first bar, when frames with three bars
were used, the bars were fixed at an equal distance
of 5 em.
On one side next to the frame holding grafted larvae
was placed frame containing pollen stores and on the
other a frame carrying young brood, for attracting
young nurse bees to the grafted larvae. These frames
on either side were then followed by frames having
emerging and sealed brood. Thereafter the colony was
copiously supplied with 60 % sugar syrup. Third day
after the introduction of the first grafts another batch
of grafted larvae was inserted in the centre of the
colony by pushing the first grafted frame aside.
From time to time the queen rearing colony was
supplemented with sealed brood from the colonies
maintained for this purpose. The graft records were
maintained by adopting the procedure of Laidlaw and
Eckert (1962). In the following proforma: date of
graft, number ofcells, breeder colony, incubator colony,
cell builder colony, number completed, date out and
remarks, if any. Three days after sealing, the queen
cells were gently removed from the wax blocks with
the help of knife and queen cells were either intro-
duced into small nuclei each for emergence or were
put into the individual queen cages within the colony
for emergence and for further observations, if any.
The queen cell cups made from bee's wax of three
different dimensions were used. The cups of each
dimension were fixed on three cell bars in a frame
and all cups prior to grafting were primed with royal
jelly. Larvae of the same age group (24 + 6 h) were
grafted in each cell cup. Each frame containing grafted
larvae was introduced into a colony of about the same
strength i.e., containing the same number of bees and
quantity of pollen and honey stores. All the three
colonies were managed uniformly during the course
of experiment.
The experiment was repeated thrice by using the
same bee colonies but the cell bars containing a
particular size of cell cups were so arranged that no
cell bar occupied the same position within the frame
more than once in order to compensate for the 10-
cational effect, if any. The number of grafts accepted
from the cell cups of each dimension i.e., the number
of queen cells sealed, was recorded and the per cent
acceptance was calculated. Grafts initially accepted
but left unattended half way were counted as rejected.
Mass rearing of Apis cerana queen 311
Material for queen cell cups
Artificial queen cell cups were made form the bees
wax obtained from old combs, fresh comb foundation
sheets and paraffin wax in the manner already des-
cribed. The size of the cell cups was kept uniform
at 6.2 and 8.6 mm internal diameter respectively at
the bees and at the mouth with depth equal to 8.8
mm. These dimensions were used in all subsequent
experiments. At one time 18 queen cell cups were
made from each material and in each frame six such
cups made of each material were fixed on each bar.
In all, nine such bars were fixed, three on each frame.
In such a way that no bar occupied the same position
in the three frames.
Suitable age of larvae for grafting
In order to ascertain the effect of larval age on the
acceptance of grafts for queen rearing and the quality
of resulting queens considered on the bases of the
volume of finished queen cells, two sets of experi-
ments were conducted. In each experiment twelve
larvae from each of the age group 12 (+6), 24 and
36 h old, were separately grafted into the queen cell
cups and fixed on the cell carrying bars. In the first
set of experiment: three bars, each carrying a different
age group of grafted larvae were fixed in a single
frame. Three such frames were prepared changing the
arrangement, so that the larvae of a particular age
group did not occupy the same position on the frame
more than once. Each such frame was introduced into
a dequeened colony. The purpose of introducing all
the three age groups of grafted larvae simultaneously
was to provide the colony ample opportunity to select
the preferred age group of larvae for queen rearing
out of the grafted larvae of three age groups.
In another experiment, twelve larvae from each age
group were grafted into the cell cups fixed on a single
bar and each bar was introduced into a dequeened
colony. The bee colonies utilized for such experiments
were devoid of all the unsealed brood so that the
colony had no option to select larvae for queen rearing
other then the ones introduced in artificial cell cups.
Both the experiments were repeated thrice and number
of grafts accepted from each age group and the volume
of all the resultant queen cells was recorded from
each colony.
Dry grafts versus grafts made over royal jelly
Effect of royal jelly on the acceptance of grafted larvae
and on the quality of resultant queens was studied
by fixing six cell cups on each cell bar. Two such
312 J. Asia-Pacific EntomoJ. Vol. 8 (2005)
cell bars were fitted into each frame. Each alternate
cell was primed with royal jelly obtained in the
manner described earlier and rest of the cell cups were
left dry. In each cell cup 12 h old larvae was
grafted. In each dequeened colony one such prepared
frame was introduced for queen rearing and number
of cells accepted from dry gra fts and grafts made on
royal jelly were recorded separately. Volume of re-
sultant queen cells so raised was also recorded for
judgment of the queens. The experiment was repli-
cated thrice.
Single versus double
For the purpose of comparing the effects of single
and double grafting on the acceptance of grafts and
the quality of queens so obtained, a batch of 24 dry
grafts, 12 on each bar were introduced into a queenless
colony. The age of the larvae was 24 (+6) h. After
two days of introduction the accepted larvae of the
same age group as grafted initially (double graft).
During the process of removal and substitution of
larvae, the consistency of the royal jelly was not
allowed to alter. In case of single grafting, the larvae
grafted initially were not disturbed and were left
intact. Data were recorded on acceptance of single
and double grafts, body length of the resultant queens
and the volume of queen cells. The experiment was
repeated thrice.
Number of grafts
Effect of number of grafts on the acceptance and
quality of queens was studied by dry grafting of
24(+6) h old larvae in batches of 24, 18, 12 and 6
grafts per frame on two cell bars. Each batch was
introduced into the colony. All the three colonies were
manipulated to be in balance with respect to bee
strength, brood area and pollen and honey stores and
their management was also kept identical. Experiment
was repeated thrice with the same bee colonies but
batches of grafts between the colonies were so
randomized that each colony got the chance to rear
a particular batch of larvae 0:11y once. Number of
grafts accepted and the volume of resultant queen cells
so raised was recorded.
Period for grafting
A batch of twelve grafts (24 + 6 hrs old larvae) was
introduced to a queenless colony each month April
to October at Bhaderwah. Bee colonies of similar
strength i.e. bees on four framers, one frame of sealed
brood, half a frame of pollen and quarter of a frame
of nectar were used to rear the grafted larvae during
the periods mentioned above. No sugar syrup was
fed to any colony during the experimental period
except a single feed of 40 % sugar syrup one day
prior to grafting. Number of grafts accepted and
volume of resultant queen cells was recorded. The
recorded data were analyzed following Snedecor and
Cochran (1967).
Results and Discussion
Besides, other factors the success of a colony depends
heavily on the qualities of a queen viz temper, in-
dustry, longevity and disease resistance etc. It, there-
fore, becomes imperative to select and rear the stock
with such qualities. The experimental data obtained
during the course ofpresent investigations is discussed
under the following heads
Queen cell cup size
The data presented in table! indicate that cell cup
size "A" was not accepted for rearing. Queen cell
cups of the size "C" recorded highest acceptance
followed by cell cup size "B". This shows that cell
cup of the size "C" are preferable to the bees as they
approximate to the natural queen cups. The degree
of acceptance and the size of the cell cups were found
to be significantly associated as revealed by Chi
square test. Given a preference to the right size, bees
are inclined to avoid queen cell cups which are
unusually smaller or larger. It could be that size "A"
is smaller to allow the proper development of the
grafted larvae and size "B" is bigger and such would
involve the wastage of material and labour to build
such giant queen cells. In the accepted, 36.1 per cent
cell cups from size "B", the bees reduce the bigger
cell cups to the proper size by coating the cups with
wax from inside, which however, seems to depend
upon the population of the young bees. Cell cup size
"c" is therefore suitable for A. cerana colonies. In
a similar study, Snelgrove (1966) considered 70 %
acceptance as good and 80 percent excellent. The
present investigations with acceptance of 75 % cell
cups of the size "C" corroborates the study of Snel-
grove (1966) who considered 70 % acceptance as good
and 80 % as excellent
Cell cup material
Out of the cell cup material tried, Paraffin cell cups
gave zero percent acceptance as against 77.8 % and
55.6 % obtained from the cell cups made from old
bees wax and wax from fresh comb foundation sheets,
respectively (Table 3). There is a significant difference
in mean volume of queen cells between other batches
of grafts made. This shows that the acceptance of
grafts was depended upon the material used for the
cell cup. The results are in line with those obtained
by Laidlew and Eckert (1962) and Ebadi and Gary
(1980) on A. mellifera.
The acceptance and volume of queen cells revealed
that batch of 12 grafts is an ideal number but where
only few queens are needed batch of 6 grafts in-
troduced into colony at a time would give better
acceptance and better qualities of queen as is evident
from the volume of the queen cells. In case more
number of queens are needed irrespective of their size
then introduction of 24 grafts at a time to a bee colony
for queen rearing could be the optimal number.
However, ifmore importance is attached to the quality
of the queens, as it should be, then batch of 12 grafts
be the optimal number. The performance can further
be improved by rearing queens in a colony with more
number of nurse bees and sufficient pollen and honey
stores. Earlier, Avetisyan et al. (1967) also found a
positive correlation between the volume of queen
cells, the weight of the queen and number of overioles
in the ovaries. Wafa and Hana (1967) also found
positive correlation between the volume of queen cells
and number of ovarioles in the ovaries. Number of
ovarioles is an index for the prolificacy of the queen.
These findings show that volume for the finished
queen cell can be taken as an index in the selection
of the better quality queens.
Mass rearing of Apis cerana queen 313
The results are in agreement with those obtained
by Doolottle (1909), who considered 12 cells as su-
fficient and 24 as too many for a colony and Snelgrove
(1966) who recommended a cell bar with 12 cells
as minimum and 24 maximum. The acceptance of
grafts was dependent upon the material used for the
construction of cell cups. The results are in full
agreement with those obtained by Laidlaw and Eckert
(1962) who advocated the use of cell cups made from
plastic were also accepted. The present findings also
corroborate the results of Ebadi and Gary (1980) who
reported that cell cups made from paraffin wax are
not accepted by bees for queen rearing but obtained
86.6% acceptance from bees wax, 70.0% from cap-
ping wax, bees wax foundation or equal parts of
paraffin and old bees wax.
From the present studies it may also be inferred
that royal jelly fad to the young larvae within the
cells imports some aroma to the combs which makes
them more acceptable to the bees. Aroma from pollen
stored into the cells also seems to make the old combs
more attractive to the bees as compared to the fresh
comb foundation sheets
Number of grafts
Batches of 6, 12, 18 and 24 larval grafts were intro-
duced into a queenless colony for rearing queens there
from. Data in table 4 exhibits highest acceptance
(72.22%) 12 and 6 grafts were introduced into a co-
lony each, followed by a batch of 18 grafts. Batch
of 24 grafts recorded lowest acceptance (48.61%).
This indicate that number of grafts introduced at a
Table 3. Effect of queen cell cup material on the acceptance of grafted larvae for rearing of A. cerana queens
Queen cup material Total number of Grafts A NA PA
Wax from old combs 36 28 8 77.8
Wax from fresh comb 36 20 16 55.6
Paraffin wax 36 0 36 0
Note: Data on total number of grafts is based on 3 colonies.
Accepted; NA = Not accepted; PA Percent accepted, X'(cal) X'(2 df, 0.05)
Table 4. Effect of number of grafts on the acceptance of grafted larvae for rearing of A. cerana queens
No. of grafts
Replication I Replication 2 Replication 3 total
introduced
A NA PA A NA PA A NA PA Graft A NA PA
24 13 II 54.16 12 12 50.0 10 14 41.66 72 35 37 48.61
18 II 7 61.11 11 7 61.11 9 9 50.0 54 31 23 57.41
12 9 3 75.0 9 3 75.0 8 4 66.66 36 26 10 72.22
6 5 1 83.33 5 2 66.66 4 2 66.66 18 13 5 72.22
Accepted; NA = Not accepted; PA = Percent accepted, X'(cal) =7.90; X
2(3
df, 0.05)
314 J. Asia-Pacific Entomol. Vol. 8 (2005)
time for queen rearing into a colony, exceeds 12,
acceptance decreases but not proportmetely. It is
further evident that there is no variation in the percent
acceptance of batch of 12 and 6 larval grafts. There-
fore, it is evident that batch of 12 grafts introduced
into a colony at a time for queen rearing is an ideal
number. Chi square test indicated the dependence of
acceptance of grafts upon the Lumber of grafts intro-
duced to a colony at a time. Taking into consideration
the volume of the resultant queen cells raised from
batches of different number of grafts, highest mean
value of 0.5567 ml was recorded for the batch of
6 grafts followed by 0.5533 ml in case of 12 grafts.
Batch of 24 grafts gave rise to queen cells, with least
mean volume (0.4967 ml) of finished queen cells.
It shows that as the number of grafts increase, the
volume of queen cells raised is reduced (Table 5).
Effect of larval age
The data in Tables 6 and 7 shows that the highest
acceptance (72.2 %) was recorded from 12 (+6) h
old grafts and lowest acceptance (24.9 %) from 36
(+6) h old grafts. The 24 (+6) h old larvae grafted
gave 63.8 % acceptance. It is, therefore evident that
younger the grafted larvae higher is the percent
acceptance. In another experiment (Table 4) where
only a single age group of larval grafts was introduced
into a bee colony, similar trend was observed. Highest
acceptance of 75.00 % was obtained from the larvae
of 12 (+6) h which was closely followed by 24 (+6)
h old larval grafts in which case 12.22 % acceptance
was recorded. Larval grafts of 36 (+6) h old gave
38.88 % acceptance which is on higher than that
obtained when all the three age groups of larval grafts
Table 5. Effect of number of grafts on volume of resultant queen cells
Number of grafts Mean volume of queen cells (ml)
Repl ication Replication 2 Replication 3 Average
24 3.50 0.49 0.50 0.4967
c
18 3.53 0.53 0.54 0.5333
b
12 3.55 0.55 0.56 0.5533
a
6 3.55 0.56 0.56 0.5567
a
Means represented by identical letters do not differ significantly.
Table 6. Simultaneous introduction of grafted larvae of 3 age groups and their effect on queen rearing of A. cerana
Age of the larvae (6hr) fotal No of Grafts A NA PA
12 36 26 10 72.22
24 36 23 13 63.88
36 36 9 27 24.99
Note: Data on total number of grafts is based on 3 colonies.
A ~ Accepted; NA ~ Not accepted; PA = Percent accepted, X'(cal) ~ 1 8 . 4 0 ; X2(2 df, 0.05) =5.99
Table 7. Effect of larval age on the acceptance of grafts and on the quality of resultant A. cerana
Colony no. Age of the grafted larvae in hr (6hr)
12 24 36
Total
A NA PA MV
Total
A NA PA MV
Total
A NA PA MV
grafts grafts grafts
I 12 8 4 66.66 0.55 12 9 3 75.00 0.54 12 5 7 41.66 0.48
2 12 9 3 75.00 0.56 12 8 4 66.66 0.53 12 4 8 33.33 0.49
3 12 10 2 83.33 0.56 12 9 3 75.00 0.54 12 5 7 41.66 0.51
Total 36 27 9 36 26 10 36 14 22
Mean 12 9 3 75.00 0.5567 12 8.3 3.3 72.22 0.5367 12 4.6 7.3 38.88 0.4930
A= Accepted; NA = Not accepted; PA ~ I'ercent accepted, MV= mean volume of queen cell (ml); X'(cal) =12.36; X'(2 df, 0.05) =5.99
Means represented by identical letters do n ot differ significantly
were simultaneously introduced into a bee colony.
Considering the mean volume of queen cells raised
from the three age groups of larval grafts it is clear
that younger the grafted larvae more is the volume
of resultant queen cells. There is a significant di-
fference in the volume of queen cells raised from
the three age groups of larval grafts. Maximum mean
volume of queen cells (0.5567 ml) was obtained from
12 (+6) h old grafted larvae followed closely by 24
(+6) h old larval grafts (0.5367 ml). These investi-
gations show that bees can rear queens from all the
three age groups of larvae tried but more preference
is given to the younger larvae. It also shows that when
a colony has no choice to select the most preferable
age group of larvae, it can rear queens in greater
number even from the higher age groups but the size
of such queens as is evident from the volume of
resultant queen cells will be smaller.
Types of grafts
The grafts made into dry cell cups and in the cell
cups primed with royal jelly (Tables 8, 9) indicated
a maximum acceptance of 77.78 % grafts made into
the artificial cell cups primed with royal jelly as
against 72.22% acceptance observed in grafts made
into dry cell cups. Ebadi and Gary (1980) obtained
93.3% acceptance for the grafts made into the cell
cups primed with royal jelly alone as against 50.9%
acceptance from the grafts made into cell cups primed
with royal jelly containing 10 % bee's stored pollen.
The variations in the percent acceptance of grafts
made into the present investigations and the obser-
vations of Ebadi and Gary (1980) could be out to
the difference in the condition of the colonies under
experimentation or due to the variations in the strength
of bees and stores present in the colony used for queen
Mass rearing of Apis cerana queen 315
rearing.
Considering the mean volume of queen cells raised
under the conditions when grafts were made into dry
cell cups and in the cell cups primed with royal jelly,
the results demonstrated that there is no difference
in the volume of queen cells raised from either type
of larval grafts. However, the present findings are
contrary to the observation of Taranov (1973) who
showed that the matter on which grafts were made
whas substantial effect on queen' s quality heaviest
queens obtained are those reared on royal jelly. His
statement is not justified on the grounds that if proper
aged larvae (12 to 24 h) are grafted there cannot be
any significant difference in the resulting queens
because such larvae would not face any feeding gap.
Even under the normal grafting technique larvae of
right age with sufficient of royal jelly present in the
worker cells is selected for grafting and the larvae
is lifted from such a cell along with royal jelly and
grafted as such to obtain good results. Under such
circumstances there is no reason to believe that larvae
of similar age grafted over royal jelly or into dry
cell cups would show variations in the body weights
of resulting queen. In addition to what has been stated
above, the process of procuring and storing of royal
jelly is not practicable keeping in view the facilities
a common beekeeper in India possess. The present
investigations are in agreement with those of Eckert
(1934) who reported that 24 hr old larvae are desirable
for grafting. Although younger larvae gave higher
acceptance and could be recommended for grafting
but are very minute and hence call for extreme care
while grafting otherwise they will easily get damaged.
The results are also is conformity with Woyke (1971)
who observed that younger the grafted larvae better
would be the resulted queens.
The variations in the acceptance of larval grafts
and volume of the resultant queen cells raised from
Table 8. Effect of type of grafts on the acceptance of grafted larvae of Apis cerana on the quality of resultant queens
Type of grafts
Grafts made over dry cell cups
Grafts made over royal jelly
A
26
28
NA
10
8
Number of grafts (total of 3 replications)
TOTAL PA Average volume of queen ell (ml)
36 72.22 0.56
36 77.78 0.56
A= Accepted; NA = Not accepted; PA Percent accepted, X'(cal) =0.4006; X'(l df, 0.05) =3.84
Table 9. Effect of number grafts on the acceptance of grafted larvae of Apis cerana on the quality of resultant queens
Type of grafts
Single grafts
Double grafts
IG
24
24
GA
16.5
16.5
PAT
71.39
71.39
A
7.3
8.0
NA
0.66
0.33
PA
91.66
95.83
MY
53.33
55.66
ML
15.63
15.77
Value of F for: mean volume of queen cells = 6.00; body length = 2.539.
IG =Initial grafted; attended; PAT=percent attended; A= accepted; NA=not accepted; acceptance; MV=mean volume of queen cells;
ML=mean linear body length of queens(mm).
316 J. Asia-Pacific Entomol. Vo.. 8 (2005)
the larval grafts of varying ages can be inferred from
the fact that in A. mellifera larvae up to first two
days of age are mass fed and afterward larvae destined
to become queens continue to be fed uninterruptedly
but worker larvae receive food after intervals. Even
the quality of the food for the two types of larvae
is changed after two days of age. This shows that
during first two days of larval life there is no di-
fference between the worker larvae and the larvae
to become queen. Therefore, if a larva is grafted up
to 2 days of age it could be readily accepted for queen
rearing as on differentiation had taken place by them.
Such a larval graft could produce good quality queens
as compared to higher age group larvae which would
have suffered a starvation gap both in quality and
quantity of food. But from the present findings it
seems that, if it is so, then 36 (+6) h old larval grafts
could have been accepted in as good number as 12
or 24 (+6) h old grafts and volume of resultant queen
cells could not have differed much. But, in earlier
studies, Morse (1979) stated hat the growth of the
ovaries of a worker larvae was considerably retarded
from the end of the first larval day in comparison
with that of a queen larvae of the same age and
possibly this could be the reason for less acceptance
of larval grafts of higher age group (above the age
of 24 h).
The difference in percent acceptance (0.56) of grafts
obtained in the present investigations between the
grafts made into cell cups primed with royal jelly
and those made into dry cell cups do not justify the
labour involved in collecting and shortage of royal
jelly.
Grafting technique
The data in Table 7 show that maximum acceptance
(95.83%) is possible by double grafts as compared
to single grafting (91.66%). The negligible difference
of 4.174% in acceptance obtained in case of single
grafting as compared to double: grafting may be due
to the disturbance caused to the nurse bees in feeding
the grafted larvae (single grafts) when the 50% of
larvae to be double grafted were being removed and
substituted with fresh grafts, two days after initial
grafting as a result of which single grafted larvae
might have faced a starvation period through very
short.
As regards mean volume of the queen cells raised
under two grafting techniques maximum volume of
queen cells (0.556 ml) was observed in case of double
grafting as against single grafting (0.543). The diffe-
rence (0.013 ml) is statistically no significant. The
average body length ofthe resulting queens for single
and double grafts recorded was 15.693 and 15.776
mm respectively. The decrease of 0.083 mm in the
body length of queens reared under single grafting
technique is also non-significant. This small variation
in the volume of queen cells and resulting queens
reared under two different conditions might have been
caused due to the short interruption in feeding the
larvae under single grafting technique, from the time
the larvae are grafted till they are fed by the nurse
bees.
Even if the differences between the two techniques
are taken cognizance of the time and labour involved
in double grafting of larvae is not justified by the
smaller difference in the acceptance and the volume
of queen cells. Again, if the larvae ofproper age which
are adequately fed before transfer are grafted, the
single grafts could give comparable results with dou-
ble grafting technique. The possibility of neglect and
inadequate feeding during short period immediately
following grafting is relatively minor compared with
skinpy feeding during the longer periods prior to
grafting (Laidlaw and Eckert, 1962).
Seasonal variability
Maximum acceptance (75.0%) of grafts was exhibited
during the month of March. This was followed by
April (66.66%), August and September (58.33% each),
May (50.00%). Lowest acceptance of 25.00% was
observed during October (Table 8). Zero acceptances
were obtained during June and July. Evidently, spring
season is the best season for queen rearing. This may
be due to the fact that during spring a tendency builds
up for more brood rearing resulting in the evolution
of swarming impulse. In a similar study, Bhat (1983)
reported the swarming season as the best season for
rearing of queens. Snelgrove (1966) and Ruttner
(1986) also highlighted the importance of weather
factors in queen rearing process.
Highest mean volume of resultant queen cells (0.55
ml) was obtained during March and August followed
by April and September (0.53 ml) and May (10.52
ml). Least mean volume (0.50 ml) was recorded
during October. These findings elucidate the point that
under Bhaderwah conditions the queen bee losses
caused during winter can be made good as early as
at the end of March by rearing queens artificially.
This will enable the colonies to develop earlier to
harvest the rich flora available during April. Similarly
for autumn requeening August is the best period for
queen rearing.
This variability in acceptance and volume for queen
cells depend upon the availability of nectar, pollen
and on atmospheric temperature. During March, April
and May major source of nectar and pollen like
Brassica, Stone fruits and other temperature fruits are
in bloom at Bhaderwah. Again during August and
September abundant pollen from maize (Zea mays)
is available whereas June and July are the dearth
periods coupled with high temperature. Rearing of
emergency queens under natural conditions during the
high atmospheric temperatures can be explained by
the fact that bees build such queen cells at the sides
of the frames and not in the centre where the tem-
perature conditions are less favorable. In producing
queens artificially the usual practice is that cells are
arranged uniformly in space between the frames
without taking into consideration the condition of
temperature in place where brood is found. Similar
observations have also been recorded by Bhat (1983)
who reported that body weight of queens varied with
season which is determined by specific conditions of
climate, honey flow, development and nutrition pro-
cess of bee colonies used in the work of queen rearing.
It is concluded that the parameters studied have
a profound influence on the mass rearing of A. cerana
queens which do not strictly follow the norms fixed
for mass rearing of A. mellifera queens. Further
studies need to be carried out to standardize the colony
condition for rearing of better quality queens.
Acknowledgment We thank Professor Yonggyun Kim,
Managing Editor JAPE and two anonymous reviewers for their
critical and very useful suggestions on an earlier draft of this
paper.
Literature Cited
Avetisyan, G.A., K.K. Bakhmatov and J.M. Zeide. 1967.
Influence of rearing periods on the external and internal
characteristics of queen honey bees. The XXIst IntI. Apic.
Congo University of Maryland, USA. 277-284 pp. Api-
mondia Publishing House Bucharest, Romania.
Bhat, A.A. 1983. Studies on queen rearing and breeding of
Apis cerana F. (Apidae: Hymenoptera). PhD. Thesis.
Himachal Pradesh Kristi Viswavidyalaya, Palampur,
Mass rearing of Apis cerana queen 317
Himachal Pradesh, India.
Ebadi, R. and N.E. Gary. 1980. Acceptance by honeybee
colonies of larvae in artificial queen cell cups. J. Apic.
Res. 19: 127-132.
Eckert, J.E. 1934. Studies in the number of ovarioles in queen
honey bees in relation to body size. J. Econ. Entomol.
27: 629-635.
Gary, N.E.1979. Activities and behaviour of honeybees. In
The hive and honeybee. Dadant and Sons, Hamilton,
Illinois.
Johansson, T.S.K. and M.P. Johansson.1978. Some important
operations in bee management. International Bee Research
Association, London.
Kither, G.V. and R.S. Pickard. 1983. Increasing the acceptance
of transplanted honeybee worker larvae by queen cell
starter colonies with the use of partially drawn artificial
queen cell cups. J. Apic, Res. 22: 175-183.
Laidlaw, RH. Jr. and J.E. Eckert. 1962. Queen Rearing. 2
nd
ed. University of California Press, Berkley. CA.
Laidlaw, H.W. 1979. Contemporary queen rearing. Dadant and
Sons, Hamilton, Illinois.
Morse, R.A. 1979. Rearing of queen honeybees. Wicwas Press
Ithaca, New York.
Ruttner, F. 1986. Geographical variability and classification.
In Bee genetics and breeding. Ed. T.E. Rinderer. Academic
press London.
Snedecor, G.W. and W.G. Cochran. 1967. Statistical methods.
Oxford & IBH Publishing Co. Ltd. New Delhi.
Snelgrove, L.E. 1966. Queen rearing. 3'd ed. Snelgrove,
Bleadon, Somerset, London.
Verma, S. and A. Sharma. 1997. Effect of various parameters
on queen rearing in Apis cerana F. in Himachal Pradesh,
India. Indian Bee J. 59: 150-153.
Wafa, A.K. and M.A. Hans. 1967. Some factors affecting the
production of royal jelly. The XXlst Inti. Apic. Congo
University of Maryland, USA. 477pp. Apimondia Publi-
shing House Bucharest, Romania.
Wongsiri, S. and S. Pthichot. 1990. Queen production and
controlled mating of Apis cerana. In Advances in in-
vertebrate reproduction. Eds. M. Hoshi and O. Yamashita.
Elsevier Science Publishers, B.V., Sweden.
Wongsiri, S., S. Pthichot and H. Fengzhi. 1988. Queen rearing
with Apis cerana in Thailand. Proc. 4
th
IntI. Conf. Bee
BioI. Research Unit Bangkok, Thailand.
Woyke, J. 1971. Correlation between the age at which honey-
bee brood was grafted, characteristics of resultant queens,
and results of insemination. J. Apic. Res. 10: 45-55.