Year-Round Production System of the Korean Native Bumblebee Bombus ignitus for Crop Pollination

Hyung Joo Yoon 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05

The bumblebee is an important pollinator of various greenhouse crops, especially for tomatoes and there has been increasing interest in commercial use of the insects for pollination. Recent advances in commercial rearing of the European bumblebee (Bombus terrestris) made it possible to package bumblebee for crop pollination. Bumblebees are distributed world widely including alpine, cool temperate and even arctic environments of the northern continents. We chose B. ignitus out of seven Korean native bumblebees, because the species showed the best results both in artificial multiplication and in pollinating ability. Now, we are studying an artificial year-round mass rearing of B. ignitus selected as the most reliable native species in crop pollination. Therefore, we investigated the optimum temperature and humidity, effect of photoperiod and CO2-treatment, facilitating effects of helper, and artificial hibernation of B. ignitus to establish year-round mass rearing of B. ignius. The experimental regimes of temperature and humidity were defined as 23℃, 27℃ and 30℃ under a constant humidity of 65% R.H., and 50%, 65% and 80% R.H. under a constant temperature of 27℃, respectively. Among the temperature regimes, 27℃-rearing showed the best results, i.e., the rates of colony initiation, colony foundation and progeny-queen production at 27℃ were 83%, 63% and 46%, respectively, which corresponded to 2.2-5.5 times the respective values at other temperature regimes. The numbers of progeny produced at 27℃-rearing, 164±33 workers, 553±174 males and 33±48 queens were also higher, corresponding to 21.8 and 1.5 times those at 23℃ and 30℃, respectively. In terms of humidity, 65% R.H. was favorable for big colony formation. Under the same humidity, the rates of colony initiation, colony foundation and progeny-queen production were 85%, 70% and 50%, respectively, and the number of progenies reached 180±30 workers, 578±179 males and 35±38 queens. Therefore, 27℃ and 65% R.H. were determined to be the favorable environmental conditions for colony development of B. ignitus in indoor rearing. It was investigated whether developmental characteristics of foundation queens of B. ignitus collected in the 4 localities in Korea would be affected by the first oviposition days of them. The first ovipostion day was classified as 1 - 4 days (immediate early), 5 - 6 days (early), 7 - 10 days (delayed early), 11 - 20 days (medium), 21 - 40 days (late), and above 41days (very late). The queen that had the early first oviposition day, i.e., laid eggs so early after starting to be raised indoors, showed much higher rate of colony foundation and progeny-queen production and much shorter period of colony foundation and worker emergence. Besides, the numbers of worker and progeny-queen emerged from the queen that had the early first oviposition day were higher than those of the queen that had the late first oviposition. In results, the queen that had the early first oviposition day could make colony stronger and could make colony formation period shorter, therefore, the first oviposition day of foundation queen was proved to be a criterion for the selection of super colonies when B. ignitus is raised indoors. It was investigated whether or not such helpers as worker bee, bee-cocoon and egg-cup etc, have any effects on oviposition and colony foundation of the bumblebee queen, B. ignitus. Among the helpers tested, the callow workers of B. ignitus and B. terrestris showed the most remarkable effects on the oviposition rates to 92% and 88%, respectively. The live cocoon as a helper improved oviposition rate over 60%. A narcotized old worker 10 days-aged after emergence, showed similar effects to a callow worker on the colony development such as oviposition rate, colony foundation and progeny-queen production. On the other hand, dried cocoon, callow honeybee worker or egg-cup did not show a positive effect as a helper. In the number of workers recruited to a foundation queen, two workers showed better effect than one worker on the colony development, with no difference between two and more. The effect of photoperiodic regimes on the oviposition and colony development of B. ignitus queens was examined with 0L, 8L, and 16L under 2 7℃ and 65% R. H. Among these photoperiod regimes, the oviposition rate at 8L and 16L was 80.2% and 83.1%, respectively, which was 12-15% higher than that at the dark condition (0L). Duration up to first oviposition at 8L and 16L was 17.5 days and 16.5 days, respectively, which was 2-3 days shorter than that at 0L. The colony foundation rate at 8L and 16L was 9.2% and 10.4%, respectively, which corresponded to 1.7-2.0 fold the value at 0L. In addition, the rate of progeny-queen production at 8L and 16L was also two fold higher than that at 0L. Taken there together, the light conditions (8L and 16L) rather than dark condition (0L) were more suitable for oviposition and colony development for B. ignitus in the indoor rearing condition. We investigated mating conditions of photoperiod, illumination and temperature during mating periods, care temperature of queen before mating, mating period and number of queen per mating cage to improve mating rate of B. ignitus. Among photoperiodic regimes of 12L, 14L and 16L during mating periods, queen mated at 14L showed better results than at 12L and 16L in egg-laying characteristics and colony development. In case of illumination during mating periods, intensity of 1000 lux was more effective than at intensity of 100 lux and 2000 lux in mating B. ignitus queen. Mating temperature and care temperature of queen before mating favorable for B. ignitus queen were 22-25? and 19?, respectively. The period need to mating B. ignitus queen was 3 days, and the number of queen suitable per mating cage of 55× 45× 65 ㎝ was 30. The effect of CO2-treatment on interrupting diapause of B. ignitus was examined to provide a means for year-round rearing of the bumblebee. When mated young queens were exposed to 65% or 99% CO2 for 30 min daily during two consecutive days, oviposition rate increased to 75% and 77%, respectively, comparing 50% in CO2-untreated queens. At the same time, the days needed to first oviposition shortened to 17-18 days in CO2-treated queens, comparing to 30 days in CO2-untreated queens. CO2-treatment at the second day after mating was appropriate to the oviposition and colony development. CO2-treatment showed a positive effect on the oviposition and colony development, but less than them of over-wintered queen in numbers of produced progeny. It can be concluded that CO2-treatment to B. ignitus is insufficient to produce commercial grade bumblebee colony in spite of its capability for promoting oviposition, because the treatment failed to form a big colony. Artificial hibernation is essential for year-round rearing of the bumblebee, B. ignitus that undergoes one generation per year. It is known that keeping the queens in low temperature for two or three months is effective to terminate their diapause and develop the colony. Temperature, time and surroundings to keep the queens during artificial hibernation were investigated. Among the tested temperatures, -2.5°C, 0°C, 2.5°C, and 5°C, the optimum temperature was 2.5°C. At the temperature (2.5°C), survival rate after chilling of the queens was high and colony development thereafter was enhanced. The proper time to initiate chilling queen was 10 to 14 days after adult eclosion, and the survivability of the queens after chilling was good during the upper period. For the surrounding to keep the queen during artificial hibernation, we proposed the method to preserve them in a bottle filled with perlite and keep it around 80% R. H.

Interspecific Cross-mating of the Bumblebees Bombus terrestris and B. ignitus

Hyung Joo Yoon,Seong Yeol Kim,Kyeong Yong Lee,Sang Beom Lee,In Gyun Park,Iksoo Kim,Byung Rae Jin 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

The large bumblebee, Bombus terrestris, which is indigenous to Europe and is used extensively for high-value crop pollination, has been artificially introduced throughout the world. Here we show the interspecific cross-mating between bumblebee species, B. terrestris and B. ignitus under laboratory conditions. The mating and oviposition rates of interspecific cross-mating of B. terrestris queen with B. ignitus male are higher than those of intraspecific matings of B. ignitus. Furthermore, the competitive copulation experiment indicates that cross-mating of a B. ignitus male with B. terrestris queen is 1.8-fold more frequent than with a B. ignitus queen. The cross-mated B. ignitus queen with B. terrestris male produced B. ignitus workers and B. ignitus male offspring, and the cross-mated B. terrestris queen with B. ignitus male produced both B. terrestris males and B. ignitus male offspring. The genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid offspring indicate that mitochondrial DNA was maternally inherited. Our results indicate that interspecific cross-mating occurred between B. ignitus and B. terrestris, which suggests that the cross-mating is not an exclusive force against the negative impact of competition and genetic contamination on native bumblebees.

Interspecific Hybridization of the Bumblebees Bombus ignitus and B. terrestris

Hyung Joo Yoon,Seong Yeol Kim,Kyeong Yong Lee,Sang Beom Lee,In Gyun Park,Ik Soo Kim 한국잠사학회 2009 International Journal of Industrial Entomology Vol.18 No.1

The large bumblebee, Bombus terrestris, indigenous to Europe and used extensively for high-value crop pollination, has been artificially introduced in several parts of the world. Here we show the interspecific hybridization between bumblebee species, B. terrestris and B. ignitus, under laboratory conditions. The mating and oviposition percentages of the interspecific hybridization of a B. terrestris queen with a B. ignites male were higher than those of the intraspecific mating of B. ignitus. Furthermore, the competitive copulation experiment indicated that the mating of B. ignitus males with B. terrestris queens was 1.8-fold more frequent than with B. ignitus queens. The interspecific hybridization of a B. ignitus queen with a B. terrestris male produced either B. ignitus workers or the B. ignitus male phenotype, and the hybridization of a B. terrestris queen with a B. ignitus male produced B. terrestris males. Genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid phenotypes indicated that mitochondrial DNA in the interspecific hybridization was maternally inherited. Our results indicated that interspecific hybridization occurred between B. ignitus and B. terrestris, which suggests that the hybridization will have a negative impact of competition and genetic pollution of native bumblebees.

Interspecific Hybridization of the Bumblebees Bombus ignitus and B. terrestris

Yoon, Hyung-Joo,Kim, Seong-Yeol,Lee, Kyeong-Yong,Lee, Sang-Beom,Park, In-Gyun,Kim, Ik-Soo Korean Society of Sericultural Science 2009 International Journal of Industrial Entomology Vol.18 No.1

The large bumblebee, Bombus terrestris, indigenous to Europe and used extensively for high-value crop pollination, has been artificially introduced in several parts of the world. Here we show the interspecific hybridization between bumblebee species, B. terrestris and B. ignitus, under laboratory conditions. The mating and oviposition percentages of the interspecific hybridization of a B. terrestris queen with a B. ignitus male were higher than those of the intraspecific mating of B. ignitus. Furthermore, the competitive copulation experiment indicated that the mating of B. ignitus males with B. terrestris queens was 1.8-fold more frequent than with B. ignitus queens. The interspecific hybridization of a B. ignitus queen with a B. terrestris male produced either B. ignitus workers or the B. ignitus male phenotype, and the hybridization of a B. terrestris queen with a B. ignitus male produced B. terrestris males. Genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid phenotypes indicated that mitochondrial DNA in the interspecific hybridization was maternally inherited. Our results indicated that interspecific hybridization occurred between B. ignitus and B. terrestris, which suggests that the hybridization will have a negative impact of competition and genetic pollution of native bumblebees.

Single Oral Dose Toxicity Study of an Alcohol Extract of Bombus ignitus pupae in Rats

Mi Young Ahn,Jea Woong Han,Hyung Ju Yoon,Jae Sam Hwang,Hae Chul Park,Yun Jung Seo,Wan Tae Chung 한국잠사학회 2009 International Journal of Industrial Entomology Vol.19 No.1

Recently, as the male silkworm pupae, bee pupae have the potential that strengths men`s vitality on vascular endothelial nitric oxide in endothelial cells. Especially we prepared alcohol extract of pupae of bumblebee, native bee named Hobakbul, Bombus ignitus. The alcohol extract of pupae of B. ignitus was administered to rats at doses of 0, 0.04, 0.2, 1 or 2 g/kg as a single oral dose. There were no observed clinical signs or deaths related to treatment in all the groups tested. Therefore, the approximate lethal dose of the alcohol extract B. ignitus pupae was considered to be higher than 2 g/kg in rats. Mild decreases in body weight gain in male were observed dose-dependently within B. ignitus pupae alcohol extract treated groups in dose response manner over 2 weeks. Throughout the administration periods, no significant changes in diet consumption, ophthalmologic findings, clinical pathology (hematology, clinical chemistry and coagulation) or gross pathology were detected. Minor changes in male and female rats were found in hematological parameters for all or partial of B. ignitus pupae extract treated groups but all the changes observed were within the physiological range. From these results, it was concluded that there was no-evidence of specific toxicity related to the ingestion of alcohol extract of B. ignitus pupae.

Single Oral Dose Toxicity Study of an Alcohol Extract of Bombus ignitus pupae in Rats

Ahn, Mi-Young,Han, Jea-Woong,Yoon, Hyung-Ju,Hwang, Jae-Sam,Park, Hae-Chul,Seo, Yun-Jung,Chung, Wan-Tae Korean Society of Sericultural Science 2009 International Journal of Industrial Entomology Vol.19 No.1

Recently, as the male silkworm pupae, bee pupae have the potential that strengths men's vitality on vascular endothelial nitric oxide in endothelial cells. Especially we prepared alcohol extract of pupae of bumblebee, native bee named Hobakbul, Bombus ignitus. The alcohol extract of pupae of B. ignitus was administered to rats at doses of 0, 0.04, 0.2, 1 or 2 g/kg as a single oral dose. There were no observed clinical signs or deaths related to treatment in all the groups tested. Therefore, the approximate lethal dose of the alcohol extract B. ignitus pupae was considered to be higher than 2 g/kg in rats. Mild decreases in body weight gain in male were observed dose-dependently within B. ignitus pupae alcohol extract treated groups in dose response manner over 2 weeks. Throughout the administration periods, no significant changes in diet consumption, ophthalmologic findings, clinical pathology (hematology, clinical chemistry and coagulation) or gross pathology were detected. Minor changes in male and female rats were found in hematological parameters for all or partial of B. ignitus pupae extract treated groups but all the changes observed were within the physiological range. From these results, it was concluded that there was no-evidence of specific toxicity related to the ingestion of alcohol extract of B. ignitus pupae.

Antibacterial Activity of Peptides Synthesized Based on the Bombus ignitus abaecin, A Novel Proline-Rich Antimicrobial Peptide

( Seong Ryul Kim ),( Eun Mi Lee ),( Hyung Joo Yoon ),( Yong Soo Choi ),( Eun Young Yun ),( Jae Sam Hwang ),( Byung Rae Jin ),( In Hee Lee ),( Ik Soo Kim ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.14 No.2

Abaecin is a largest member of the proline-rich antimicrobial peptide family found only in the hymenopterans. A cDNA of abaecin was previously isolated and cloned from Bombus ignitus: the mature peptide of Bombus ignitus abaecin was composed of 39 amino acid residues. In the present study, we determined the antibacterial effect of B. ignitus abaecin synthesized at several lengths against several bacteria by radial diffusion assay. The 37-mer peptide (Ab37) inhibited the growth of Gram-negative bacteria Escherichia coli ML-35, Pseudomonas aeruginosa and Salmonela typhimurium, but showed limited inhibitory activity toward Gram-positive bacteria, except for Micrococcus luteus. The truncated 26-mer peptide (Ab26), which was synthesized after truncating some amino acid residues at both N-terminus and C-terminus from the Ab37 peptide, still showed equivalent antibacterial activity to the Ab37. On the other hand, several further truncated peptides exhibited lower activity then did Ab37 peptide.

Mating Conditions Favorable for Improving Mating Rate of the Bumblebee, Bombus ignitus

( Hyung Joo Yoon ),( Sam Eun Kim ),( Kyeong Yong Lee ),( Sang Beom ),( In Gyun Park ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.15 No.2

We investigated mating conditions of photoperiod, illumination and temperature during mating periods, care temperature of queen before mating, mating period and number of queen per mating cage to improve mating rate of Bombus ignitus. Among photoperiodic regimes of 12L, 14L and 16L during mating periods, queen mated at 14L showed better results than at 12L and 16L in egg-laying characteristics and colony development. In case of illumination during mating periods, intensity of 1000 lux was more effective than at intensity of 100 lux and 2000 lux in mating B. ignitus queen. Mating temperature and care temperature of queen before mating favorable for B. ignitus queen were 22-25℃ and 19℃, respectively. The period need to mating B. ignitus queen was 3 days, and the number of queen suitable per mating cage of 55×45×65㎝ was 30.

Isolation of Insect pathogenic bacteria from Bombus terrestris and B. ignitus in Korea

Won Tae Kim,Byeong Do Park,Ji Young Choi,Jong Gill Kim,Seok Jo Whang,Hyung Joo Yoon,Sang Mi Han,Mi Ae Kim 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

Bumblebees are important pollinators in greenhouse and have colonized all parts of the World. In Korea, the value of bumblebees is increasing as pollinator. However, the more recent use of reared colonies may ultimately allow pathogens to spread to peripheral areas for bumblebees. Generally, bumblebees are hosts to a large number of parasites which are mites, flies, protozoa, fungi, virus and bacteria. For most of these, very little is known about their effects against host insect, epidemiology or evolutionary ecology. So, we report pathogenic bacteria isolated from Bombus terrestris and B. ignitus at first time in Korea. Bacillus fusiformis and Klebsiella oxytoca are isolated from B. terrestris and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of B. fusiformis was 35~40% against B. terrestris. Pantoea dispersa and K. oxytoca are isolated from B. ignitus and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of these species were 35~40% against B. ignitus. These pathogenicity are considered as low-level.

Mating Conditions Favorable for Improving Mating Rate of the Bumblebee, Bombus ignitus

Yoon, Hyung-Joo,Kim, Sam-Eun,Lee, Kyeong-Yong,Lee, Sang-Beom,Park, In-Gyun Korean Society of Sericultural Science 2007 International Journal of Industrial Entomology Vol.15 No.2

We investigated mating conditions of photoperiod, illumination and temperature during mating periods, care temperature of queen before mating, mating period and number of queen per mating cage to improve mating rate of Bombus ignitus. Among photoperiodic regimes of 12L, 14L and 16L during mating periods, queen mated at 14L showed better results than at 12L and 16L in egg-laying characteristics and colony development. In case of illumination during mating periods, intensity of 1000 lux was more effective than at intensity of 100 lux and 2000 lux in mating B. ignitus queen. Mating temperature and care temperature of queen before mating favorable for B. ignitus queen were $22-25^{\circ}C$ and $19^{\circ}C$, respectively. The period need to mating B. ignitus queen was 3 days, and the number of queen suitable per mating cage of $55{\times}45{\time}65\;cm$ was 30.