Nosema disease caused by microsporidia which belongs to fungi is a major cause of honey production loss and is worldwide in distribution. The most common method of nosema disease controlling is the application of the fumagillin. It is an antibiotic derived from the fungus Aspergillus fumigates. It has been used for many years to treat microsporidiosis caused by Nosema in honeybees. However, Fumagillin does not kill Nosema spores, and has rapidly deteriorating potency after application, resulting in only partial and temporary anti-Nosema effect, since new bees emerge constantly in a colony, and re-application is required several times a year. Also it is no longer practised for the associated risk of honey contamination. In this study we compare the effect in spore reduction of Cordyceps militaris extract(CMX) and of fumagillin. Three weekly administrations of CMX showed a noticeable decrease in the number of spores infecting the honey bee intestines, without negative effects for adults and comb health. Therefore this product could play an important role in prevention and reduction of nosema disease without risks of honey with antibiotics.

This study was conducted to establish the optimized protocol for cytoplasm isolation of bee pollen. Data of biochemical parameters and amino acid profiles were obtained from acorn pollen grains treated with pulverization or lyophilization. Contents of moisture, ash, crude protein and crude fat of acorn pollen were 11.7%, 2.6%, 24.1% and 11.8%, respectively. After pulverizing, content of crude protein was decreased to 23.8% while crude fat was 22.5% which means 90% increase. Also content of crude protein was increased to 26.5% in case of the lyophilized pollen. Amino acids such as aspartic acid, glutamic acid, leucine and arginine were extensively found in acorn pollen while histidine, methionine and cystine were infrequent. The pulverized pollen was increased by 2.6% in the total amino acid percentage while the lyophilized pollen increased by 11.8% compared to the untreated pollen.

Most Korean beekeepers have moved from south to north of Korea to collect nectar from black locust (Robinia pseudoacacia) flowers for 2 months. This provided a valuable opportunity to sample bees originating from diverse areas in one location. We initiated a survey of honeybee (Apis mellifera) colonies on the blooming period of Acacia to determine the prevalence of Nosema apis and black queen cell virus (BQCV) in 2013. Nosema causes significant losses in population size of honeybees. Sixteenth hives were sampled for this study. Bees were collected on the 4th and 13th of May, 2013. Nosema spore counts ranged from zero to 1,948,333 spores per bee. The average number of nosema spores per bee was calculated to be 450,000. Approximately 94% of the apiaries examined were infected with nosema, based on the presence of spores in the flowering period of Acacia. Also nosema is thought to be associated with black queen cell virus. RT-PCR analysis shows that BQCV infection rate was 100%. This indicates that nosema and BQCV is the predominant species affecting honeybee colonies.

Chungbuk province has number of management honey bee(Apis cerana) colonies. A. cerana produce honey, and are pollinators with in agricultural crops and natural environmental ecosystems. Korean Sacbrood Virus(SBV) causes colony collapses of A. cerana Feb. in Korean apiaries from 2009 years. It causes a fatal disease(sacbrood) of honeybee larvae, which fail to pupate, change color and shape, and finally die. We thus conducted a molecular survey of honey bee RNA viruses, Nosema microsporidia, Foul broods and fungal disease(Chalk brood and Stone brood) associated with environmental conditions in apiaries and differential type(Traditional and Modern) of A. cerana hives in Chungbuk province. We found the presence of Israel Acute Paralysis Virus(IAPV), Nosema apis, and Sacbrood virus(SBV) was detected in the tested samples. IAPV were detected from mordern hive. Nosema apis, and Sacbrood virus(SBV) was detected from traditional hives. Our results demonstrate that honey bee RNA viruse(SBV) and N. apis are present in Traditional hives. It can suggest SBV and other disease might be related environment conditions(Type of hives).

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 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.

Nosema disease caused by Nosema apis which belongs to fungi is a major cause of honey production loss and is worldwide in distribution. N. apis infects the epithelial cells of the digestive system of adult honeybees. Nosema causes significant losses in population size of honeybee. There are about 25 thousand beekeepers caring for approximately 1,697,000 colonies in Korea. Honey production totaled almost 38,505 metric tons in 2010. This production was estimated to be worth about 274 billion Korean won. To determine infection level of nosema disease during the season, adult worker bees were collected from two colonies of experiment apiary from January to October. Our results indicate that the infection level of nosema disease was increased in spring and autumn. Also we initiated a survey of honeybee colonies on the blooming period of Acacia to determine the prevalence of N. apis. Twenty two hives owned by 18 beekeepers were sampled for this study. Bees were collected on 24th and 25th May of 2012. Nosema spore counts ranged from zero to 5,266,000 spores per bee. The average number of nosema spores per bee was calculated to be 1,375,000. Approximately 86% of the apiaries examined were infected with nosema, based on the presence of spores in the flowering period of Acacia. This indicates that nosema is the predominant species affecting honeybee colonies.

Honeybees (Apis mellifera) adapted themselves to different geographical and climatical conditions since they had been introduced in Korea. Beekeepers have tried to breed valuable lineages with artificial insemination or conventional mating techniques. However, evaluation of breeding resultants still relies on timeconsuming observation data. Genetical characterization of breeds has proven its usefulness to preserve genetic resources of livestock. In recent years, microsatellites are most commonly used to evaluate population structures and diversities of living organisms in that the characteristics of locus specificity, rich polymorphism, abundant and random distribution over the genome, and their co-dominant inheritance. Determining classic genetic distances using neutral, highly polymorphic microsatellite markers is a reliable method to investigate genetic relationships and breed differentiation. This methodology can be used to establish preservation priorities for livestock breeds. The final aim of this study was to develop potent markers for assessing genetic structure of lineages after artificial insemination. In this study, the genetic structure of ten microsatellite markers were sequenced or analysed with polymerase chain reaction for eleven European honeybee populations. The results may help to develop reliable microsatellite markers for more efficient preservation strategies of valuable honeybee breeds.

The average life expectancy has continuously increased with the development of medicine. However, Some of men have been suffered from the insufficient secretion of testosterone causing by physical factors, social and psychological factors. Testosterone is an essential steroid hormone controlling male reproductive function. Alternative medicines in plants, fungi, and insects have been studied to enhance sexuality. In the present study, we determined the effect of an acorn pollen and Cordyceps militaris fruiting body on testosterone concentration in Sprague-Dawley rats. Eighteen rats per group were housed to regular diet or diet supplemented with acorn pollen powder or fruiting body power for 4 weeks. Serum was collected from 6 rats per group. Results showed that changes of the body weight, food and water intake of the rats were not observed in this study. However, acorn pollen powder and Cordyceps militaris fruiting body significantly stimulated testosterone production (p < 0.05) compared to the control, respectively. Hence, it suggests that acorn pollen or Cordyceps militaris fruiting body might be developed as a complementary medicine to improve sexual hormones.

Cordyceps militaris (CM) has been used as a traditional folk medicine to strengthen the immune system and regain energy in Far East Asian countries. Some of men have been suffered from the insufficient secretion of testosterone causing by physical factors, social and psychological factors. Testosterone is an essential steroid hormone controlling male reproductive function. Alternative medicines in plants, fungi, and insects have been studied to enhance sexuality. Cordyceps species including Cordyceps sinensis and C. militaris has been used as for the enhancement of sexual function for hundreds of years. In the present study, we determined the effect of fruiting bodies of C. militaris which cultured on bee drone medium (DCM) on testosterone concentration in Sprague-Dawley rats. Eighteen rats per group were housed to regular diet or diet supplemented with DCM for 4 weeks. Serum was collected from 6 rats per group. Results showed that changes of the body weight, food and water intake of the rats were not observed in this study. However, DCM significantly stimulated testosterone production (p < 0.05) compared to the control. Hence, it suggests that fruiting bodies of C. militaris grown on the bee drone medium might be developed as a complementary medicine to improve sexual hormones.

Two species entomopathogenic fungi most widely used and valued in traditional Asian medical practice are Ophiocordyceps sinensis (formely Cordyceps sinensis) and Cordyceps militaris. Although O. sinensis may be the more famous and expensive fungus, it is also comparatively rare and cannot be grown or made to fruit readily in culture whereas C. militaris occurs worldwide, can be easily cultured, and is the easist of all Cordyceps species to fruit in culture. There is a well establihed cottage industry in Korea to produce C. militaris as a dietary supplement or even as a culinary ingredient used to promote improved health. Most of the Korean farmers raising C. militaris obtain fruiting bodies from silkworms that are injected with suspensions of hyphal bodies grown in liquid cultures. This study seeks to facilitate and to simplify the injection process used to produced this fungus by finding a simple cullture medium on which abundant supplies of the Lecanicillium conodial state of C. militaris are produced and can be used with simplified injection protocols involving spraying or dipping in conidial suspensions than the more material- and labor- intensive injection protocol. The studies to be repored include quantitative tests of conodial yields on varying carbon sources, varying nitrogen sources, and attempted to optimize the carbon/nitrogen ratio and pH of the medium for conidial production.

This study aimed to evaluate the antioxidant activity of chestnut honey which were harvested at various areas in South Korea. First at all, we measured the total phenols content through a spectrophotometric determination with a modified Folin-Ciocalteu method and total flavonoids content determined with aluminium chloride. Total phenolic compounds was highest in Sunchang of Chestnut honey(2.21mg/ml)and flavonoids contents was also the highest in Sunchang of Chestnut honey(1.02mg/ml) than other samples. For measured the antioxidant activity of chestnut honey, we performed DPPH(2,2-diphenyl-1-picrylhydrazyl) test and FRAP(ferric reducing-antioxidant assay)test. DPPH scavenging activity highest in Sunchang of Chestnut honey more than 50% DPPH scavenging activitywhile other samples (Gong-ju, Yechen, Chung-ju, Imsil, Ha-dong) showed more than 25% DPPH scavenging activity. The ferric reducing-antioxidant assay (FRAP) is based on the reduction of ferric 2,4,6-tris(2-pyridyl)-1,3,5-triazine [Fe(III)-TPTZ] by spectrophotometric analysis. Sunchang were found to have more than 532μM FRAP activity while other samples (Gong-ju, Yechen, Chung-ju, Imsil, Ha-dong) showed more than 300μM FRAP activity. The results suggested that chestnut honey strong antioxidant activity and it could be utilized as a source of natural antioxidant.

We report for the first time the occurrence of DWV-infected bumble bees (Bombus ignitus). For the present study, the detection of DWV virus from the female and male bumble bee was investigated in the same colony. The Deformed wing virus (DWV) of honeybee (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. Using specific RT-PCR protocols for the detection of DWV followed by sequencing of the PCR products we could demonstrate that the bumble bees were indeed infected with DWV. The virus was detected from Bombus ignitus, and its partial DWV gene was cloned and sequenced. The partial DWV gene encoding the polyprotein is 711-nt of 235 amino acid residues. The deduced nucleotide sequence of the polyprotein partial gene of DWV showed 96.9%, 96.2%, 96.8%, and 96.5% homology to other structure polyprotein partial gene of DWV from insects, respectively. Phylogenetic analysis further conformed that the deduced nucleotide sequence of the polyprotein partial gene of DWV divided to the outside tree. We describe the first time that presence of Deformed wing virus(DWV) from bumble bee(Bombus terrestris) in korea using RT-PCR.

The traditional use of insects as food continues to be widespread in tropical and subtropical countries and to provide significant nutritional, economic and ecological benefits for rural communities. Specially, Bee brood serves as a food source to humans in many countries although limited data exists concerning its nutrient composition. Bee brood (pupa and larvae) were analyzed for Carbohydrate, Saturated fatty acid, Cholesterol, protein, fat, fiber, minerals, and vitamins. Bee brood was high in protein(46.4%～46.73%), fat(18.84%～ 20.75%),carbohydrate(24.66 %～35.79 %), Folic acid(222.30 ㎍/100g), and vitamins. Differentially, folic acid had been contained by high density in pupa of drone. While low in iron, bee brood was a good source of folic acid, and carbohydrate. The fat was composed mostly of saturated and mono-unsaturated fatty acids. The present data suggest bee brood to be an excellent source of many valuable nutrients including energy, amino acids, many essential minerals, and B-vitamins. These data suggest bee brood could be a valuable source of nutrients to various populations.

The caterpillar-shaped Chinese medicinal mushroom (DongChongXiaCao) looks like a worm in the winter and likes a grass in the summer. The fruiting bodies of Cordyceps species have demonstrated various biological activities and used as traditional medicines in Asian countries. The price of wild Cordyceps have been ascending every year as their demand increase. Recently, the artificial cultivation of these fungi has been successfully established and they are currently produced on a rice or pupae media. This study was carried out to investigate the soluble sugar, amino acid and fatty acid profiles in the fruiting-bodies of Cordyceps militaris, C. pruinosa and Paecilomyces. tenuipes formed on the living silkworm (Bombyx mori) host. Soluble sugars such as glycerol, glucose, mannitol and sucrose were found in the fruiting bodies of Cordyceps militaris, C. pruinosa and P. tenuipes. Total soluble sugar content was 29.23 ㎎g-1in C. militaris, 8.61㎎g-1 in C. pruinosa and 24.00 ㎎g-1in P. tenuipes on dry weight basis. Total free amino acid content was 14.09 ㎎g-1in C. militaris, 34.60㎎g-1 in C. pruinosa and 17.09 ㎎g-1 in P. tenuipes. The content of oleic acid in fatty acids was above high more than 30% regardless of species.

경제적 부가 가치가 높은 붉은자루동충하초의 안정적 대량생산 기틀을 마련하고자 자실체 증식 특성 및 자실체 성분 분석을 한 결과를 요약하면 다음과 같다. 1. 붉은자루동충하초의 감염률은 관행인 누에 5령 기잠시분무 접종하는 방법보다 번데기 시기에 주사접종 하는 방법이 높았다. 2. 붉은자루동충하초의 자실체 발생은 온도 21∼25℃, 습도 91% 이상, 광 1500 lx 이상에서 양호하였다. 3. 붉은자루동충하초의 유리당 함량은 Glucose가 7.9 ㎎으로 가장 많았으며, 다음이 Sucrose 이었으며 Glycerol과 Mannitol은 미량 검출되었다 4. 붉은자루동충하초의 누에 자실체에 함유된 아미노산의 총 함량은 34.6 μmole/g이며, 16종의 구성 아미노산중 Arginine이 15.6%로 가장 높았으며, Lysine이 3.4%로 가장 낮게 분포하였다. 5. 붉은자루동충하초의 누에 자실체 유리 지방산 조성은 Oleic acid 함량이 가장 많았으며, Palmitoeic acid는 소량 함유되어 있었다. 6. 붉은자루동충하초의 누에 자실체 유기산 조성은 Citric acid가 가장 많았으며, Malic acid와 Propionic acid는 미량 존재하였다.