This is the first record of endoparasitic Hymenoptera Apanteles galleriae recorded from Apis cerana colonies in Korea. A simple rearing protocol was established to allow the morphology, mating behavior and infestation rate of A galleriae. In total, 55 lesser wax moth fresh cocoons were kept in the tissue culture test plate at room temperature (25.6 ± 1.5˚C, RH 21 ± 3.7%). The females were 3.4 ± 0.3 longer than male 3.3 ± 0.2. The male antenna was longer than females. The copulation lasts 24.4 ± 2.4 seconds. The larvae of A. galleriae were pupated inside the cocoons of lesser wax moth. Ninety percent of adults A. galleriae was successfully emerged from the lesser wax moth cocoons. A. galleriae can be used as bio-logical control in store and in live colonies to control lesser wax moth.

We investigated the duration of laying worker oviposition and egg-laying behavior in three queenless colonies of A. cerana by in situ video recording. Egg load was determined by dissecting laying workers in September 2012. Egg size, length and breadth, shape index and egg elongation were calculated. To determine the number of eggs laid by laying workers per cell at 24, 53, 74, 120 and 171 hours was also monitored. To estimate the number of eggs per cell per week, a small comb was squeezed between two frames. The combs were collected at given hours and weekly to count the number of eggs, respectively. The results showed that the duration of oviposition of laying workers on average was 109.2 ± 67.5 seconds per cell. During oviposition, egg-laying workers showed two types of behaviors, viz; a still phase, where the egg-laying workers did not move, and a recovery phase, where the egg-laying workers vigorously wagged their abdomens after oviposition. The results showed that on average, 4.0 ± 5.1 of worker eggs per cell per week was recorded. The highest number of eggs was recorded at 120 hours compare to at 24, 53, 74 and 171 hours. Three different shapes of the eggs namely oval, elongated long and elongated curve shaped was laid by workers. The results showed that the laying worker carried 1 to 4 mature eggs in her ovaries and may lay from one to four per oviposition. In conclusion, the laying worker shows a still and a recovery phase during and after laying the eggs. The laying workers retain 1 to 4 eggs in their ovaries. The breadth of eggs is strong positive relationships with length. One worker cell can accumulate up to 33 eggs in queenless colonies.

Ovarioles are smooth, gradually widening white tubes with different stages of eggs. The ovarioles were gently removed, and the right and left ovarioles were separated and counted the ovarioles. We observed that the ovaries of laying queens were extended from second abdominal up to fourth abdominal segments. Each ovariole is supplied with tracheae. The tracheae are auriferous types characterized by coating spiracle tubules with permeable cuticle, which may bring the tracheal air into close contact with haemolymph.

We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

In the present study, the 17,694-bp long complete mitochondrial genome (mitogenome) of the dwarf honey bee, Apis florea (Hymenoptera: Apidae), is described with an emphasis on the noteworthy triplicated tRNAser(AGN) region and an extraordinary long A+T-rich region with repeat regions. The gene arrangement of A. florea mitogenome is identical to that of A. mellifera, but has triplicated tRNASer(AGN), each of which contains the precedent 44 bp-long and following another 64 bp-long repeats plus one complete first repeat abutting to tRNAMet. A total of 1,610-bp long two repeat regions in 1,987 bp-long A+T-rich region is composed of nearly identical 141 ~ 219-bp long five tandem repeats and 50 ~ 52-bp long 12 tandem repeats that are encompassed by three non-repeat sequences. One of the common interpretations for such repeat sequence is slipped-strand mispairing and unequal crossing-over events during DNA replication.

We compared the grafting success in total of 107 rearing Apis carana queens cells, to which we grafted 540 larvae. The wax for cups we prepared from A. mellifera and A. cerana wax. The A. cerana wax cups were found that artificial queen cell cups with the internal diameter of 8.0 mm at the mouth and 8.0 mm depth were highly preferred by the bees for rearing of queens from the grafted larvae. From the 210 grafted larvae into A. mellifera wax bees accepted 30 queens cells, only (16.67 %) ; A. cerana wax bees accepted 59 queens cells (33 %) ; plastic cup bees accepted 18 queens cells, only (10 %). In the preference test the grafting success in the A. cerana wax cups were better than in the A. mellifera wax and plastic cup. The results show better acceptance of larvae grafted into the pure A. cerana wax cups for rearing A. cerana queen. A new method for rearing honey bees, A. cerana, in vivo was developed and the effects of royal jelly from A. mellifera. We used royal jelly diluted 50:50 with sterile water (The royal jelly is kept frozen until used). A small amount of royal jelly is placed at the center of each cell cup. Young A. cerana larvae were transferred into the queen cups containing ± 10 ㎍ of the Royal jelly from A. mellifera and A. cerana. The average rates of acceptance were affected significantly due to the royal jelly source in the queen cell cups. It is so workable first to produce pure A. cerana wax for making the queen cups before a beekeeper starts with grafting.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.