PROPAGATION OF Apis cerana FABR.

(HYMENOPTERA: APIDAE)
PROSPECTIVE QUEEN BEE

Jasmi Jasmi1*, Syarifuddin Syarifuddin2 & Dewirman Prima Putra3

1
Biology Study Program, STKIP Sumatera Barat, Jl.
Gunung Pangilun Padang, Indonesia.
2
Biology Study Program, FMIPA,
State University of Medan, Medan, Indonesia.
3
Technology of Agricultural Product,
Ekasakti University, Padang, Indonesia.
*Corresponding author: [Link]@[Link]

ABSTRACT

Honey bee, Apis cerana Fabr. (Hymenoptera; Apidae) has been domesticated in West Sumatra.
Various attempts to obtain practical and effective methods for multiplying superior colonies
have been made. Optimizing the role of worker bees as a determinant of the future colony can
be done by creating an emergency colony condition (queen bee not present). In this kind of
colony, the worker bees immediately produced a new queen bee candidate from young larvae.
This research was conducted at Trigona Mandiri Apiary, Padang Pariaman, West Sumatra,
Indonesia from February to May 2019. The method used was a complete randomized design
with four treatments and 10 replications. Results showed that the comb isolation technique was
the best method for producing new queen bee candidates and increasing the appearance of
young queen bee (gyn). The mean appearance of gyn from pupae was 4.8 per colony so that
the number and quality of pupae to be maintained can be selected.

Key words: Apis cerana, worker, queen cell, emergency colony, colony multiplication.

ABSTRAK

Lebah madu, Apis cerana Fabr. (Hymenoptera; Apidae) telah dibiakkan di Sumatera Barat.
Pelbagai usaha telah dilakukan untuk mendapatkan kaedah yang praktikal dan efektif dalam
meningkatkan koloni-koloni unggul yang dipelihara. Mengoptimalkan penanan lebah pekerja
sebagai penentu masa depan koloni dapat dilakukan dengan menciptakan koloni emergency
(ratu lebah tidak hadir). Dalam koloni sebegini, lebah pekerja segera menghasilkan calon ratu
lebah baru dari larva muda. Kajian ini telah dijalankan dari bulan Februari hingga Mei 2019 di
Apiari Trigona Mandiri, Padang Pariaman, Sumatera Barat. Kaedah yang digunakan adalah
melalui rekabentuk rawak dengan empat perlakuan dan 10 ulangan. Dari hasil kajian, diketahui
bahawa teknik isolasi comb adalah kaedah terbaik untuk menghasilkan calon ratu lebah baru
serta kemunculan ratu lebah muda (gyn). Rata-rata kemunculan gyn dari pupa adalah sebanyak
4.8 per koloni sehingga dapat dilakukan pemilihan terhadap jumlah dan kualiti pupa yang akan
dipelihara.
Serangga 25(2): 80-95 Jasmi et al.

Kata kunci: Apis cerana, lebah pekerja, sel ratu, koloni emergency, perbanyakan koloni

INTRODUCTION

Honeybees are a group of social insects that form caste structures with specific functions in
their colonies. The composition of the castes are queen bee, sterile female bees (worker bees)
and male bees (Michener 1974; Wilson 1971). Each caste has distinct duties and functions so
it cannot be replaced by a different caste. The division of duties in honey bee colonies is the
most studied behavioral phenomenon. Research on the honey bees behavior has investigated
since the 1800s, but serious experimental research only started in the early 1930s and continues
until today with various laboratory approaches based on various biological perspectives
(Beekman et al. 2007; Schmickl & Crailsheim 2007; Smith et al. 2008). This is because worker
bees play an important role in determining the future of the colony such as preparing hives,
food foraging even changing the sex of the larvae. The process of changing the sex of the larvae
is carried out by worker bees if the colony is in an emergency condition that occurs if the colony
lost the queen bee (Ribbands 1953).

Queen bee is the most important individual in a honeybee colony, because only queen
bee that can produces both fertile and sterile eggs and produces pheromones to prevent worker
bees from developing new queens and ovaries (Delaney et al. 2011; Winston 1987). Good
quality queen bee is a fundamental factor for successful apiculture practices. A high-quality
queen bee of Apis mellifera can be obtained by various methods such as isolation of queen bee
(Tarpy et al. 2016), moving young larvae (12-24 hours old) from worker cells to queen bee
cells that hung vertically in the same hive (Büchler et al. 2013), the Doolittle grafting method
with wax and plastic bowls and utilization of the Cupkit and Karl Jenter apparatus (Dhaliwal
et al. 2017). From those previous studies, the easy and practical methods were relatively few.

Apis cerana has an important role as a pollinator, especially for agricultural crops and
orchards (Partap 2011; Sung et al. 2006) such as coffee plantation (Klein et al. 2003; Saepudin
et al. 2011), Jathropha curcas (Atmowidi et al. 2008), Mangifera indica L. (Deuri et al. 2018)
and Prunus armeniaca (Gurmani et al. 2016). In West Sumatra, this honeybee is easily found
in lowland farming until altitude 1400m above sea level. Aside from being a pollinator, A.
cerana also cultivated for producing honey. In West Sumatra, especially Padang Pariaman
region, the cultivation of A. cerana was established from 1984 to the present. Honeybee colony
are obtained by moving the wild colony to beehives by using the hiving method (Jasmi et al.
2014). The continuous poaching of wild A. cerana colony is directly affecting the presence and
balance of insect’s pollinator (Erniwati & Kahono 2009). Other factor that affect the wild A.
cerana colony is the increase of wasp population, due to the wasp’s predatory behavior (Arif
et al. 2014; Omran et al. 2011).

Three common conditions in the natural maintenance of queen bee are loss of queen
(resulted in an emergency colony), supersedure cells and swarming (Wade 2014). The principle
of an emergency colony can be utilized to produce prospective queen bee. The manipulation
of emergency conditions in superior colonies is very useful in producing new colonies that
have the same relative qualities as the original colonies. Emergency conditions in bee colonies
can be manipulated by various methods such as isolating the queen bee in A. mellifera (Tarpy
et al. 2016), isolating combs (Büchler et al. 2013; Maramis & Rompas 2015; Tofilski &
Czekonska 2004) and dividing the colony in Tetragonula carbonaria (Nunes et al. 2014). In
addition to the emergency colony, queen bee production can also be stimulated by cutting the
top of the combs (Puspitaningrum 1995; Snelgrove 1981). This study was focused on testing

ISSN 1394-5130 81
Serangga 25(2): 80-95 Jasmi et al.

the effectiveness of the isolation and comb cutting method in producing A. cerana queen bee
cells.

MATERIALS AND METHODS

This study was conducted at Apiari Trigona Mandiri located in Kanagarian Batu Gadang
Kuranji Hulu, Sungai Geringging District, Padang Pariaman, West Sumatra, Indonesia from
February - May 2019. Geographically, the study site is located at 100’ 07’ 00” East longitude
and 0’ 33’00” South latitude (BPS 2018). The colonies were maintained by Apiary Trigona
Mandiri originated from wild Apis cerana colonies that were transferred to the beehive. Colony
age was varied from five to 30 months, but the colony used for the sample was ranged from
five to six months. This study used 4 treatments and 10 replicates, thus a total of 40 colonies
were involved. The treatments were comb isolation (A), comb-cutting with a triangle pattern
(B), comb-cutting with a horizontal pattern (C) and control (D). The comb isolation method
used refers to Büchler et al. (2013) whereas comb cutting following to Snelgrove (1981) and
Stahlman (2013). Beehive size used in this study refers to Schouten et al. (2019) which is 40 x
30 x 20 cm. A beehive has nine frames with the area of each frame is 32 x 16 cm (Figure 2).

The colonies used in this study must be in a good condition, have exactly eight combs
with a comb area in each frame more than 256 cm2 or >50% of the total frame space and at
least there are three frames containing young brood cells (white-colored comb). If a colony has
nine combs, one of the combs (preferably the broken or old one) was cut and the rest were put
back into the beehive. All frames containing the combs were examined to ensure that there are
no queen bee cells in all combs of the test colony. The results of beehive sorting were labeled
on-site.

The treatment of comb insulation was done by placing the isolator between the 4th and
th
5 frames. The isolator was made from cardboard sized 35 x 26 x 0.3 cm (Figure 1). In each
beehive there was nine frames (eight frames containing a comb and one empty frame). The
isolator was divided and isolated the frame into two groups with four frames on the left (1st to
4th frame, along with the combs and adult bee occupying it) and five frame on the right (5th to
9th frame, along with the combs and adult bees occupying it and one empty frame) (Figure 2A
& 2B). The model and size of the isolator was following the model and size of the beehive so
that the isolator can play a role mainly to isolate the transfer of the queen bee from the left
frame-group to the right frame-group or vice versa. After the isolator was installed, the beehive
was closed.

Figure 1. Model of comb isolation device used for propagation of Apis cerana queen bee

ISSN 1394-5130 82
Serangga 25(2): 80-95 Jasmi et al.

A B

C D
Figure 2. The methods used to produces Apis cerana queen bee. (A) The isolator was
located between the 4th and 5th frame, (B) the placement of the isolator between
the 4th or 5th frame which already contains the comb, (C) comb with horizontal
cutting pattern and (D) comb with triangle cutting pattern.

The frame chosen for the comb cutting method was the one with a minimum 75% comb
area and have young combs which were marked with white or brownish wax. This process was
carried out on 20 colonies by cutting the comb edge with two patterns. A total of 10 colonies
were cut in a triangular pattern, 10 colonies horizontally and 10 other colonies were used as
controls. The triangle pattern was made by cutting the comb from the top part to form the peak
of the triangle in the middle of the comb at 50% height from the bottom of the frame (Figure
2D). The comb area cut following the triangle pattern was approximately 64 cm2. Horizontal
pattern was made by cutting the ¼ part from the comb edge (Figure 2C) or cutting the m-p area
(Figure 3).

ISSN 1394-5130 83
Serangga 25(2): 80-95 Jasmi et al.

Figure 3. The used frame model to estimate the increase in comb area of Apis cerana. a-
p = sub-areas in a frame with an size of 32cm2, equivalent to 6.25%

Examination of the prospective queen bee cells were carried out on each beehive comb
for all treatment colonies on the 18th day. For the isolated colonies, the dismantling of isolator
was done if the prospective queen cells were formed. All queen bee cells were allowed to
develop naturally. An observation of the emerged pupae of prospective queen bee was done on
the 24th day. The emergence of prospective queen bee pupae was marked by the abandoned
pupae case.

Measured Parameters
Number of prospective queen bee cells
The number of prospective queen bee cells produced by each colony was calculated by
checking the entire comb at the end of the observation (day 24th). The number of prospective
queen bee cells counted was only normal queen bee cells (emergency cells and supersedure
cells were not counted).

The total of emerged queen bee cells

Emerged queen bee cells were characterized by an open-top and empty cells. The number of
emerged queen bee cells was determined for each comb per colony, following this formula,

𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑒𝑚𝑒𝑟𝑔𝑒𝑑 𝑞𝑢𝑒𝑒𝑛 𝑏𝑒𝑒 𝑐𝑒𝑙𝑙𝑠

= 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑝𝑟𝑜𝑠𝑝𝑒𝑐𝑡𝑖𝑣𝑒 𝑞𝑢𝑒𝑒𝑛 𝑏𝑒𝑒 𝑐𝑒𝑙𝑙𝑠 – 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑒𝑚𝑒𝑟𝑔𝑒𝑑 𝑐𝑒𝑙𝑙𝑠

Area of the comb

The size of the used frame was 32 x 16 cm. To estimate the area of the comb formed in the
frame, a frame model was made by dividing the frame area into 16 sub-area with size 32 cm2
each (Figure 3). The model frame was attached to the frame containing the comb. The comb
addition was calculated from the area of the newly formed comb on the frame from the cut
comb and blank frames. Comb area (%) was estimated in the frame at the end of the observation

ISSN 1394-5130 84
Serangga 25(2): 80-95 Jasmi et al.

based on Delaplane et al. (2013) criteria by modifying the area of the frame into sub-areas
(Figure 3).

Data Analysis
The treatment effects towards the total of prospective queen bee were analyzed using the one-
way ANOVA Test (Analysis of Variance) followed by a post-hoc test using the Duncan test.
The comb area size was expressed in % and analyzed using the chi-square formula. All analyses
were performed using SPSS software version 21. Data for total emerged queen bee was
expressed in percent (%) following the formula below,

Total emerged prospective queen bee

Percentage of the emerged prospective queen (%)= x100%
Total of normal queen bee cells

RESULTS AND DISCUSSIONS

ANOVA results showed that there were differences between treatments on the comb size area
and the average number of prospective queen bee cells formed from each sample colony (P
<0.05). Duncan’s analysis results (α = 0.05) showed that the largest comb average increase
within a 18 days was found on the control colony, while the least was found on triangle pattern
comb cutting treatment (Table 1). The highest average number of queen bee cells produced
was found in the isolated treatment comb, while the least was found in comb cutting treatment.

Table 1. The results of Duncan's analysis for an average increase in the area of Apis
cerana (cm2) combs and the number of prospective queen bee cells over a period
of 18 days
Measured Parameters
Total
Treatment Number of prospective queen
Samples
Comb area (cm2) bee (cell)
Comb isolation 10 38.40a 6.80b
Horizontal comb cutting 10 69.60b 0.00a
Triangle comb cutting 10 72.00 b 0.00a
No treatment (control) 10 133.60c 0.50a
* Different letters that indicate differences between treatments

Comb isolation is a method of dividing one bee colony into two groups, where one of the
group has no queen bee, with the expectation to produce two independent colonies. The part of
the colony without a queen bee is known as an emergency condition. The loss of the queen bee
is a serious threat to the sustainability of the honeybee colony. In such condition, worker bees
will play a reproductive-related role for the survival of the colony (Dixon et al. 2014). Worker
bees plays an important role during the rearing phase of the larvae of prospective queen bee
(AL-Kahtani 2011). According to Brouwers et al. (1987) during the stages of larval
development, the worker bees will provide different food (in quality and quantity) for the cells
of the prospective queen bee and worker bees (royal jelly vs worker jelly). This condition
induced different development processes in these two cell types. As explained by Linksvayer
et al. (2011), the control of nutritional factors is very important to larvae development, which
is proven by the disappearance of dimorphism between the queen and the worker if the nutrients
given to the larvae are the same.

ISSN 1394-5130 85
Serangga 25(2): 80-95 Jasmi et al.

The number of colonies produced the prospective queen bee cells were only 10 of 40
colonies, those were eight colonies from the comb isolation method and two colonies from
control (Table 2). From 10 colonies that produced prospective queen bee cells, as many as
seven colonies produced normal prospective queen bee cells, three colonies produced abnormal
prospective queen bee cells and two colonies produced both types of cells. The number of
frames occupied by prospective queen bee cells was only 26 out of the total 90 frames used.
Out of these 26 frames, 17 frames are occupied by normal prospective queen bee cells, three
frames are occupied by abnormal prospective queen bee cells and six frames have both types
of cells.

Table 2. Number of colonies that produce queen bee cells and combs are occupied by
Apis cerana queen bee cells after 18 days of observation
Total comb occupied
Total Total colony produced
by prospective queen
Methods sample prospective queen bee cells
bee cells
colony
Normal Abnormal Normal Abnormal
Triangle cutting pattern 10 0 0 0 0
Horizontal cutting pattern 10 0 0 0 0
Comb isolation 10 8 5 23 6
No treatment (control) 10 2 0 3 0
Total 40 10 5 26 6
Average 10 2.5 1.75 7.00 1.50

The propagation of the queen bee in Apis cerana was more effective with the comb
isolation method (Table 2). The comb isolation method created an emergency condition for
some part of the colony, then worker bees will immediately take action to produces prospective
queen bee. Shi et al. (2011) stated that the emergency colony only had six days to produce a
new queen after the last egg was produced by the old queen. If six day were passed, the egg
has developed into a larva and loses its ability to develop into a functional queen (eggs hatch
after 3 days and larvae under the age of 3 days still have totipotential ability).

As mentioned in the previous paragraph, the time required to produce A. cerana

prospective queen bee in an emergency colony was relatively short. Emergency conditions will
affect the behavior of worker bees in determining the future of the colony by producing new
prospective queen bee. Büchler et al. (2013) reported that the success and quality of queen bee
production depend on the strength and adequacy of the nutrition of a colony, as well as the
quality of colony management. The important thing in raising queen bees for emergency
colonies according to Woodward (2007) is the process of transferring 12-24 hour old larvae
from worker cells to queen cells located vertically in the middle of the hive. These larvae will
be fed royal jelly by worker bees. Royal jelly is a high-quality food source that can change the
sex of the bee while it is still in the larval phase for less than three days. Rafique et al. (2019)
reported that the addition of royal jelly in prospective queen bee cells before the new larvae
grafted was more effective compared to the addition during the queen rearing process.

The distribution of normal queen bee cells were on frames 1–8 (Table 3). A total of
eight colonies separated by an insulator (8 parts on the left and 8 parts on the right) were
produced normal queen bee cells. All parts of the colony that were separated by an insulator
can produce prospective queen bee cells both from the part of the colony (with or without queen

ISSN 1394-5130 86
Serangga 25(2): 80-95 Jasmi et al.

bee). The total number of normal prospective queen bee cells was higher in the frame located
to the left side of the insulator (46 cells) compared to the right side (22 cells).

Table 3. Distribution of normal queen bee cells found on each comb of 10 Apis cerana
colonies on 18th observation day
Comb (frame) number per total prospective queen
Colony Total
Method bee cells
code
1 2 3 4 5 6 7 8 9
Comb 1 0 3 3 4 3 1 0 1 0 15
isolation 2 0 3 2 4 2 2 0 0 0 13
3 0 0 1 2 1 0 0 0 0 4
6 0 1 8 2 3 1 2 0 0 17
7 0 1 3 0 1 0 0 0 0 5
8 1 0 2 1 1 0 0 0 0 5
9 0 2 1 2 0 1 0 0 0 6
10 0 0 0 0 0 2 1 0 0 3
Summary 1 10 20 15 11 7 3 1 0 68
Average 0.22 2.22 4.44 3.33 2.44 1.56 0.67 0.22 0.00 15.11
Control 5 0 0 0 1 0 1 0 0 0 2
8 0 1 0 0 2 0 0 0 0 3
Summary 0 1 0 1 2 1 0 0 0 5
Average 0.00 0.50 0.00 0.50 1.00 0.50 0.00 0.00 0.00 2.50
Total 1 11 20 17 13 8 3 1 0 73

The highest number of normal prospective queen bee cells was found in the 3rd frame
(20 cells) and 4th frame (15 cells), while no cells found on the 9th frame. The highest number
of normal queen bee cells in one frame was 10 cells which were consisted of eight normal cells
and two abnormal cells (Figure 4b). The highest number of normal prospective queen bee cells
was found in the comb isolation method which was 7 cells per frame. The highest number of
normal queen bee cells produced by one colony was 17 cells per colony.

Prospective queen bee cells were placed in all the comb area categories (Table 4). The
average number of prospective queen bee cells produced was 18.25±33.25 cells per treatment
or 1.82±0.83 cells per colony. The largest number of queen bee cells was found in the comb
area >384 cm2 (comb area covers >75% of frame space) which was 41 cells and at least in the
comb area less than 128 cm2 (comb area covers <25% of frame space) which was 2 cell. The
wider the comb in the isolation treatment, the more number of queen bee cells were found. For
the comb cutting treatment no queen bee cells were produced within the 18 days observation
period.

ISSN 1394-5130 87
Serangga 25(2): 80-95 Jasmi et al.

Table 4. Number of prospective queen bee cells found in various groups of comb area
from 10 Apis cerana colonies after 18 days of treatment
Comb area (cm2) per
total number of prospective queen bee Total cells of
Method cells prospective
256- queen bee
>384 129-256 <128
384
Triangle cutting pattern 0 0 0 0 0
Horizontal cutting pattern 0 0 0 0 0
Comb isolation 37 23 6 2 68
No treatment (control) 4 1 0 0 5
Total comb 41 24 6 2 73
Stdev for treatment 17.93 11.34 3.00 1.00 33.25
Average per colony 1.02 0.6 0.15 0.05 1.82
Stdev per colony 0.45 0.28 0.08 0.03 0.83

The comb structure occupied by the prospective queen bee cells was varied. Prospective
queen bee cells were placed on a comb that occupied by brood cells, food storage and empty
cells (Figure 4). Some queen bee cells were also placed on the same comb as the worker and
drone/ male bee brood cells (Figure 4a and 4b), drone brood cells only (Figure 4d) and worker
bee brood cells only (Figure 4c). Prospective queen bee cells were also found on the same comb
as food storage cells containing pollen and honey (Figure 4a) and pollen only (Figure 4d).
There are also queen bee cells that are placed on a comb with only empty cells (Figure 4c).

Figure 4. Some area and cells conditions of Apis cerana combs occupied by prospective
queen bee cells after 18 days of observation. (a) The comb area was >384 cm2
occupied by prospective queen bee, worker bees and male bees brood cells, food

ISSN 1394-5130 88
Serangga 25(2): 80-95 Jasmi et al.

storage and empty cells. (b) The comb area was 256-384 cm2 occupied by
prospective queen bee, worker bees and male bees brood cells, food reserves
and empty cells. (c) The comb area was 129-256 cm2 occupied by prospective
queen bees and worker bee brood cells also empty cells. (d) The comb area was
<128 cm2 occupied by prospective queen bees brood cells, pollen cells and
empty cells.
1 = queen cell, 2 = empty cell, 3 = worker pupa cell, 4 = drone pupae cell, 5 =
honey cell, 6 = pollen cell.

The Total Prospective Queen Bee Cells that Emerged as New Queen Bee (Gyn)
The total number of prospective queen bees emerged as young queen bee (gyn) on the 24th day
was 71.23% which consisted of 65.75% from comb isolation treatment and 5.48% from control
colonies (Table 5). The success of emerged pupae to gyn was higher (100%) in colonies with
fewer queen bee cells. A higher percentage of non-emerged pupae (>30%) was found in
colonies that had more than 5 prospective queen bee cells.

Table 5. Nmber of Apis cerana prospective queen bee cells that emerged on the 24th
observation day
Total emerged pupae of prospective queen bee/
Colony code / colony
Method
number Percentage of (%)
Pupae Emerged
emergence
Comb isolation 1 15 11 73.33
2 13 9 69.23
3 4 3 75.00
6 17 11 64.70
7 5 3 60.00
8 5 4 80.00
9 6 4 66.66
10 3 3 100.00
Summary 68 48 588.92
Average 8,5 6 73.62
Stdev 5.55 3.66 12.37
No treatment 5 2 2 100.00
(control) 8 3 2 66.66
Summary 5 4 166.66
Average 2.5 2 83.33
Stdev 0.71 0.00 23.57
Total 73 52 71.23

The percentage of emerged pupae using the comb isolation method was 65.75% (Table
5). The success of the young bees’ emergence will vary for each method and type of bee, for
example in Apis mellifera with cupkit queen rearing technique the percentage of emergence
was 54.67% (Dhaliwal et al. 2017) whereas Doolittle grafting method the queen bee emergence
percentage was 53% and 77% (Emsen et al. 2003). The usage of in-vitro rearing method with

ISSN 1394-5130 89
Serangga 25(2): 80-95 Jasmi et al.

the addition of a highly concentrated sugar solution also able to increase the number of the
prospective queen bee (Kaftanoglu et al. 2011).

The new queen bees were found at 18th observation day in a colony which had 17 cells
of prospective queen bee cells (Figure 5). Prospective queen bee that emerged as gyn was
marked by the opening of the lid (cup) at the top of the pupae cell. The wings of newly emerged
queen bee appeared a bit tangled but slowly the wings will dry and look tidy. Prospective queen
bee cells that were failed to emerge were contained of dead pupae in damaged condition.

Figure 5. Newly emerged young queen bee (gyn) of Apis cerana on the colony with an
initial comb of nine frames

The comb isolation method is more effective to produce a high number of prospective
queen bee cells (Table 5). One colony can produce as many as 17 prospective queen bee cells,
where in one comb 8 normal prospective queen bee cells can be found (Figure 5). The
emergence of relatively high number of prospective queen bee in a single colony can be caused
by various factors such as colonies in emergency conditions (Tarpy et al. 2016; Tofilski &
Czekonska 2004) and artificial prospective queen bee cells through various methods (AL-

ISSN 1394-5130 90
Serangga 25(2): 80-95 Jasmi et al.

Kahtani 2011; Dhaliwal et al. 2017; Emsen et al. 2003; Kuntadi 2013; Okuyan & Akyol 2018;
Rafique et al. 2019).

CONCLUSION

In this study the comb isolation technique was the best method for producing new prospective
queen bees and to increasing the appearance of young queen bees. Comb isolation is an
excellent technique for the apiary with difficulties in practicing colony propagation. The
average appearance of young queen bees (gyn) from pupae was 4.8 per colony so that the
number and quality of pupae to be maintained can be selected.

ACKNOWLEDGEMENTS

We would like to thank the General of Research and Technology of Higher Education for
funding this research through the PTUPT GRANT 2019. Our gratitude conveyed to Ajo Lapau,
Ramadhani Saputra as a field data collector assistant, the head of the Trigona Mandiri Apiary
group, Batu Gadang community and Pakandangan Padang Pariaman who have provided
coconut plantation facilities for placement of beehives in this research.

ISSN 1394-5130 91
Serangga 25(2): 80-95 Jasmi et al.

REFERENCES

AL-Kahtani, S. N. S. 2011. Selection of larvae for queen rearing by workers in the honey bee
(Apis mellifera L.) − Nepotism or different maternal supply? Dissertation, Humboldt-
Universität zu Berlin.

Arif, M. M. J., Norashirene, M. J., Khairiyah, M. H. S., Nicholas, A. S., Aman, A. Z., Abdul-
Latiff, M. A. B., Aifat, N. R., Yaakop, S. & Yusof, M. D. F. Z. 2014. Initial study of
social vespid wasps by using molecular approach of 28s region. Serangga 19(1): 51-
61.

Atmowidi, T., Riyanti, P. & Sutrisna, A. 2008. Pollination effectiveness of Apis cerana
Fabricus and Apis mellifera Linnaeus (Hymenoptera: Apidae) in Jathropha curcas L.
(Euphorbiaceae). Biotropia 15(2): 129-134.

Beekman, M., Gilchrist, A. L., Duncan, M. & Sumpter, D. J. 2007. What makes a honeybee
scout? Behavioral Ecology and Sociobiology 61(7): 985-995.

BPS [Badan Pusat Statistik]. 2018. Statistics of West Sumatra in 2018. Padang: The central
statistics agency of west sumatra province [Indonesia].

Brouwers, E. V. M., Ebert, R. & Beetsma, J. 1987. Behavioural and physiological aspects of
nurse bees in relation to the composition of larval food during caste differentiation in
the honeybee. Journal of Apicultural Research 26(1): 11-23.

Büchler, R., Andonov, S., Bienefeld, K., Costa, C., Hatjina, F., Kezic, N., Kryger, P., Spivak,
M., Uzunov, A. & Wilde, J. 2013. Standard methods for rearing and selection of
Apis mellifera Queens. Journal of Apicultural Research 52(1): 1-30.

Delaney, D. A., Keller, J. J., Caren, J. R. & Tarpy, D. R. 2011. The physical, insemination, and
reproductive quality of honey bee queens (Apis mellifera L.). Apidologie 42(1): 1-13.

Delaplane, K. S., Van Der Steen, J. & Guzman, E. 2013. Standard Methods for estimating
strength parameters of Apis mellifera colonies. Journal of Apicultural Research 52(1):1-
12.

Deuri, A., Rahman, A., Gogoi, J., Borah, P. & Bathari, M. 2018. Pollinator diversity and effect
of Apis cerana F. pollination on yield of mango (Mangifera indica). Journal of
Entomology and Zoology Studies 6(5): 957-961.

Dhaliwal, N. K., Singh, J. & Chhuneja, P. K. 2017. Comparative evaluation of Doolittle, Cupkit
and Karl Jenter techniques for rearing Apis mellifera Linnaeus queen bees during
breeding season. Journal of Applied and Natural Science 9(3): 1658 -1661.

Dixon, L. R., Kuster, R. D. & Rueppell, O. 2014. Reproduction, social behavior, and aging
trajectories in honey bee workers. AGE 36(1): 89-101.

Emsen, B., Dodologlu, A., Genc, F., Smith, B. H. & Cobey, S. 2003. Effect of larvae age and
grafting method on the larvae accepted rate and height of sealed queen cell (Apis
rnellifera L.). Journal of Applied Animal Research 24 (2): 201-206.

ISSN 1394-5130 92
Serangga 25(2): 80-95 Jasmi et al.

Erniwati & Kahono, S. 2009. The role of wild plants in the conservation of pollinating insect
order Hymenoptera. Jurnal Teknik Lingkungan 10 (2): 195-203. [Indonesia]

Gurmani, H. T. W., Jaleel, W., Saeed, S., Gurmani, A. R., Corresp, Q. S., Naqqash, M. N.,
Ghouri, F., Munawar, M. S., Sial, M. U., Rahman, S. & Aine, Q. U. 2016. The
pollination potential of Apis cerana Fab., Apis mellifera L. (Hymenoptera: Apidae) on
yield of apricot (Prunus armeniaca). Peer Journal Preprints 1-19.

Jasmi, Salmah, S. & Dahelmi. 2014. Nesting site Apis cerana Fabr. (Hymenoptera: Apidae) in
two different altitude of polycultureplantations in West Sumatra. Hayati Journal of
Bioscience 21 (3): 135-143.

Kaftanoglu, O., Linksvayer, T. A. & Page, R. E. 2011. Rearing honey bees, Apis mellifera, in
vitro 1: effects of sugar concentrations on survival and development. Journal of Insect
Science 11(96): 1-12.

Klein, A-M., Steffan-Dewenter, I. & Tscharntke, T. 2003. Bee pollination and fruit set of
Coffea arabica and C. canephora (Rubiaceae). American Journal of Botany 90 (1):
153-157.

Kuntadi. 2013. Effect of larval ages on the quality of Apis cerana L.(Hymenoptera: Apidae)
queens breed in captivity using queen-grafting technique. Jurnal Entomologi Indonesia
10 (1): 1-6. [Indonesia].

Linksvayer, T. A., Kaftanoglu, O., Akyol, E., Blatch, S., Amdam, G. V. & Page Jr, R. E. 2011.
Larval and nurse worker control of developmental plasticity and the evolution of honey
bee queen-worker dimorphism. Journal of Evolutionary Biology 24(9): 1939-1948.

Maramis, R. T. D. & Rompas, J. J. I. 2015. Utilizing the queen rearing method of Apis mellifera
for beekeping development. Zootek 35 (2): 235-246. [Indonesia].

Michener, C. D. 1974. The Social Behavior of the Bees a Comparative Study. Cambridge.
Massachusetts: The Belknap Press of Havard University.

Nunes, T. M., Heard, T. A., Venturieri, G. C. & Oldroyd, B. P. 2014. Emergency queens in
Tetragonula carbonaria (Smith, 1854) (Hymenoptera: Apidae: Meliponini). Austral
Entomology 54(2): 154-158.

Okuyan, S. & Akyol, E. 2018. The effects of age and number of grafted larvae on some
physical characteristics of queen bees and acceptance rate of queen bee cell. Turkish
Journal of Agriculture - Food Science and Technology 6(11): 1556-1561.

Partap, U. 2011. The pollination role of honeybees. In Hepburn, H. R. & Radloff, S. E. (eds.)
Honeybees of Asia, pp. 227-255. Heidelberg: Springer-Verlag Berlin.

Puspitaningrum, W. I. 1995. The effect of comb cutting on the number of queen cells built by
Apis cerana honeybees. Undergraduate Thesis. FMIPA. Universitas Diponegoro.
Semarang. [Indonesia]

ISSN 1394-5130 93
Serangga 25(2): 80-95 Jasmi et al.

Omran, N. S. M., Hussein, M. H., Khodairy, M. M. & Awad, A. M. 2011. Predators of

honeybee and its impact on activities of honeybee colonies under conditions of South
Valley, Egypt. Journal Agricultural and Biological Science 7(1): 79-88.

Rafique, M. K., Mahmood, R., Qadir, Z. A., Bodlah, I. & Shaheen, F. A. 2019. Effects of
rearing interlude and grafting technique on honeybee Apis mellifera L. queen under
field conditions. Pakistan Journal of Zoology 51(6): 1-4.

Ribbands, C. R. 1953. The Behavior and Social Life of Honey Bees. London: Bee Res. Assn.

Saepudin, R., Fuah, A. M. & Sumantri, C. 2011. Increased productivity of honeybee through
the application integration with coffee plantations. Jurnal Ilmu-Ilmu Peternakan 21: 29-
39. [Indonesia]

Schmickl, T. & Crailsheim, K. 2007. HoPoMo: A model of honeybee intracolonial population

dynamics and resource management. Ecological Modelling 204(1-2): 219-245.

Schouten, C., Lloyd, D. & Lloyd, H. 2019. Beekeeping with the asian honey bee (Apis cerana
javana Fabr) in the Indonesian islands of Java, Bali, Nusa Penida, and Sumbawa.
Bee World 1-5.

Shi, Y. Y., Huang, Z. Y., Zeng, Z. J., Wang, Z. L., Wu, X. B. & Yan, W. Y. 2011. Diet and
cell size both affect queen-worker differentiation through DNA methylation in honey
bees (Apis mellifera, Apidae). PLoS ONE 6(4): e18808.

Smith, C. R., Toth, A. L., Suarez, A.V. & Robinson, G. E. 2008. Genetic and genomic analyses
of the division of labour in insect societies. Nature Review Genetics 9:735-748.

Snelgrove, L. E. 1981. Queen Rearing. 4th Edition. Bleadon Hill, Weston-Super Mare, Avon:
Snelgrove & Smith.

Stahlman, D. 2013. A Queen Manual. London: Department of Agriculture Division of Plant

Industry- Apiary.

Sung, I. H., Lin, M. Y., Chang, C. H., Cheng, A. S. & Che, W. S. 2006. Pollinator and their
behaviors on mango flowers in Southern Taiwan. Formosan Entomol 26: 161-170.

Tarpy, D. R., Simone-Finstrom, M. & Linksvayer, T. A. 2016. Honey bee colonies regulate
queen reproductive traits by controlling which queens survive to adulthood. Insectes
Sociaux 63(1): 169-174.

Tofilski, A. & Czekonska. K, 2004. Emergency queen rearing in honeybee colonies with brood
of known age. Apidologie 35(3): 275-282.

Wade, A. 2014. Queen Rearing and Use of Queens. Cambera: Beekeepers Association of the
ACT (Department of Primary Industries).

Wei, S., Raina, S. K. & Fries, I. 2001. Colony development and queen rearing in Kenyan
honeybees (Apis mellifera scutellata). Durban, South Africa. Proc. 37th Int. Apic.
Congr. 28 Oct-1 Nov 2001.

ISSN 1394-5130 94
Serangga 25(2): 80-95 Jasmi et al.

Wilson, E. O. 1971. The Insect Societies. Cambridge: Harvard University Press.

Winston, M. L. 1987. The Biology of the Honey Bee. Cambridge: Harvard University Press.

Woodward, D. 2007. Queen Bee: Biology, Reading and Breeding. New Zealand: Balclutha.

ISSN 1394-5130 95