본 연구는 LMO 유전산물의 위해성평가를 위해 바실러스로부터 증폭된 Vip3Aa 유전자를 이용하여 대장균에서 단백질 순수분리 하였으며, MALDI-TOP 분석법을 통해 기존의 알려진 살충성 Vip3Aa 단백질과 동등한 단백질임을 증명하였다. 순수 분리한 Vip3Aa 단백질을 이용하여 꿀벌 과독성 급성섭식독성평가를 수행하였다. 그 결과 무처리군, Hepes buffer, Vip3Aa 단백질 처리군 모두 치사 및 일반 중독증상을 보이는 개체는 발견되지 않았다. 이 결과를 통해 Vip3Aa 단백질은 꿀벌에 위해성을 나타내지 않는다는 결론을 얻을 수 있었다. 본 연구 결과는 향후 국내 LMO 유전자산물 위해성평가에 유용하게 활용될 것이라 사료 된다.

낭충봉아부패병에 저항성인 신품종 토종벌을 농업과학원에서 시험재료로 제공받아 일반 농가에서 사육중인 일반벌과 비교하였다. 꿀벌 수밀력을 조사한 결과 일벌 개체당 수밀력은 비저항성 계통보다 저항성계통이 15% 높았으며, 화분채집량은 저항성계통에서는 358mg, 비저항성 계통은 330.3mg으로 저항성계통이 비저항성계통보다 8.4%의 높은 화분채집력을 보였다. 질병저항성을 알아보기 위해 봉개유충 청소행동 평가에서 8시간 후 저항성계통에 서는 51.3%, 비저항성계통에서는 50.7%이었으며, 24시간 후에는 두계통 모두 98.7~100%의 사충 제거율을 보였다. 봉군발육조사에서는 저항성 계통의 경우, 일벌, 유충, 번데기의 수가 비저항성계통에 비하여 41%, 7%, 73% 로 발육이 우수하였다.

Here, we investigated antioxidant defense mechanism in the spermatheca of A. mellifera queens via RNA-seq analysis of spermathecae in both mated and virgin queens. We identified the genes encoding antioxidant proteins, which were differentially expressed in the spermatheca of mated queens. The concentrations of antioxidant proteins, such as superoxide dismutase 1 (SOD1), catalase, glutathione peroxidase (GTPX), and transferrin (Tf) together with the levels of ROS, H2O2, and iron were higher in the spermathecal fluid of mated queens as opposed to those in the spermathecal fluid of virgin queens; this indicated that increase in antioxidant protein concentration is an antioxidant defense mechanism occurring in the spermathecal fluid of mated queens against ROS; this mechanism involves conversion of ROS using antioxidant enzymes and Tf-mediated inhibition of the Fenton reaction occurring between Fe2+ and H2O2. Our data indicate that an increased expression of antioxidant proteins could facilitate prolonged storage and survival of sperms in the spermatheca of mated queens, suggesting the role of antioxidant proteins in antioxidative defense against ROS.

We investigated the effects of honey bee (Apis mellifera L.) colony size on the pollination of greenhouse-cultivated watermelon grown under the forcing cultivation system. The highest pollination activity of bees was observed (14.3 ± 5.0 honey bees/day) when the bee colony size was 10,000 honey bees, followed by 7,500 and 5,000 honey bees. There was a positive correlation between the bee colony size and pollination activity (R = 0.262). There was no significant difference in the fruit set with different honey bee colony sizes (88%–91%). However, the larger the bee colony size, the higher the number of seeds fertilized and the rate of seed fertilization (p > 0.05). The number of seeds and the content of sugar were negatively correlated (R = -0.714). Overall, we found that a colony size of 5,000 honey bees was the most effective for the pollination of watermelon grown under forcing cultivation.

Learning and memory are essential for animals like honey bee, in which foragers leave their nest to collect food and then successfully learn to navigate back to colony. Chronic environmental exposure of neoniconitiod highly affect pollinators like honey bee. We investigated the effect of neonicotinoid, thiacloprid and imidacloprid on learning, memory and mortality of pollen foragers based on their toxicity level. Individual Pollen foragers Apis mellifera were trained using a proboscis extension proboscis. We used three treatments: 50% sugar solution, 50% sugar contaminated thiacloprid and imidacloprid as unconditioned stimulus and 2M 1-nonanol as conditioned stimulus. Our result shows that there is no significant difference between control and thiacloprid treatment both in learning and mortality of honey bee. But we found difference in memory between subjects exposed to control and thiacloprid contamination. Whereas imidacloprid contamination significantly lowered learning, eradicate memory retention and higher mortality of honey bee. Even thiacloprid and imidacloprid are in the same chemical class of Neonicotinoid insecticides, their toxic level would be different resulting in differential ecological impact to honey bee colony.

Comparative toxicity of different groups of pesticides on Apis mellifera L. were tested in the laboratory. Commercial formulations dissolved in water from the highest dose level marked on the label were serially diluted to 10^(-6) times to carry out oral trials on each pesticide. Fungicide (Metconazole), herbicide (Glyphosate), acaricide (Amitraz) and 3 insecticides (organophosphate Fenitrothion, neonicotinoids of Thiacloprid and Thiamethoxam) were compared. Thiamethoxam and fenitrothion caused higher mortality, and amitraz showed intermediate level of mortality while herbicide and fungicide showed minimal impacts. Among neonicotinoids, Thiamethoxam was highly toxic but thiacloprid showed mortality less than 25% even at the highest concentration. LC50 values were estimated as Amitraz 197.70, Fenitrothion 2.20, and Thiamethoxam 0.188 ppm at 24 h after treatment. Fungicide, herbicide and a neonicotinoid thiacloprid were all showed low oral toxicity. This study confirmed that the old pesticide, organophosphate was highly toxic and two different toxic path way exist for neonicotinoid pesticides.

기후변화, 외래병해충발생, 밀원수 감소 등 여러 요인으로 인해 우리 나라의 양봉농가는 벌꿀생산량 및 농가소득이 감소하는 추세이다. 또한 밀원이 없는 시기에 가축용 사료가 아닌 고가의 식용설탕을 꿀벌의 인공사료로 급여하고 있어 농가는 ‘생산비 상승’과 ‘밀원수 부족’을 국내양봉업이 직면한 가장 큰 문제점으로 지적하고 있다. 이에 따른 양봉농가의 생산비를 절감하고 꿀벌의 생리·생태의 개선효과를 가지는 저가형 당액사료개발을 위해 농산부산물에서 추출한 천연당을 봉군에 적용시험을 추진하였다. 양파(OS-1, OS-2), 양배추(CS-1, CS-2), 바나나(Banana)부산물에서 추출한 천연당의 봉군당 평균먹이소모량(mL)은 설탕당액(대조구) 처리구와 비교시 바나나추출당액 급여구에서 5500.00±0.00, 5133.33±404.15로 유사하였다. 그러나 꿀벌의 봉세발달은 대조구에 비해 21006.7±2137, 17403.3±2257마리로 조사되어 낮은 경향을 나타내었고 수명조사 결과는 큰 차이가 없었다. 농산부산물을 이용한 천연당은 우선 꿀벌의 기호도 향상을 우선으로 개선시켜야 할 것으로 보인다.

To control an external parasitic mite, a honey bee line possessing high hygienic behavior (HHB) against an external parasitic mite, Varroa destructor, has been bred in South Korea and an assessment method has been necessitated to diagnose HHB line from the low hygienic behavior (LHB) line. Thus, in this study, we developed single nucleotide polymorphism (SNP) markers from whole genome sequencing of each 20 worker bees from HHB and LHB lines of A. mellifera ligustica (Hymenoptera: Apidae). An average of 319,445,977 sequence reads was mapped to the known A. mellifera reference genome (an average of 87.46%). In 2,316,128 and 3,266,756 SNPs from each HHB and LHB line, an average of 93.6% and was located in the intergenic spacers and introns, whereas, the remaining 6.4% was located in the genic region, respectively. Among them 20 SNPs that were fixed at each line possessing within-individual homozygosity were selected and each four SNPs were used to diagnose the two honey bee lines either by typical PCR-restriction fragment length polymorphism method or allele-specific PCR. The remaining six SNPs had the size difference, enabling relatively easy differentiation between the two honey bee lines on typical agarose gel and another remaining six SNPs only has sequence difference including SNP sites. Thus, these SNP markers can be used to diagnose the honey bee line with HHB from LHB line against V. destructor.

Insect peptides have been extensively studied due to beneficial effects in the treatment of infectious diseases. Melittin, a fundamental component of honeybee venom produced by European honeybee Apis mellifera, has applied to prevent various inflammatory disease and bacterial infections in human. However, the therapeutic application of melittin is limited due to its low stability, hemolytic activity and expensive manufacturing costs. In this study, we aimed to discovery unknown peptides from the Apis mellifera and evaluate its antibacterial activity against Escherichia coli KACC 10005. A total 15,853 peptide sequences were diciphered using Illumina HiSeq 2500 next-generation sequencing (NGS) platform and analyzed based on the Apis mellifera official Gene Set Version 3.2 (amel_OGSv3.2) and the Collection of Anti-Microbial Peptides (CAMPR3) database. All the peptide sequences and annotation data sets were combined and sorted by physicochemical features of antimicrobial peptides (AMPs), such as short peptide length <=50, positive charge, isoelectric point (8.0<=pl<=12), and aggregation propensity (in-vitro: <=500, in-vivo: –40<= Na4vSS <=60). Among the screened peptides, four unknown peptide candidates, named AMP1-4, were chemically synthesized and tested for antimicrobial activity in comparison with a reference peptide, melittin. Inhibition of bacterial growth was observed in the AMP4 treated group from 6 hours to 48 hours post-treatment against E. coli. These results suggest that honeybee-derived peptide sequences can be applied as natural resources to acquire novel AMPs and the peptide sequences derived parameters are enough to recognize antibacterial peptides. In addition, the selected novel peptide candidate, AMP4, has antibacterial activity.