Honey bees are crucial pollinators for agricultural and natural ecosystems, but are experiencing heavy mortality in Korea due to a complex suite of factors. Extreme winter losses of honey bee colonies are a major threat to beekeeping but the combinations of factors underlying colony loss remain debatable. Finding solutions involves knowing the factors associated with high loss rates. To investigate whether loss rates are related to Varroa control and climate condition, we surveyed beekeepers in korea after wintering (2021–2022 to 2022–2023). The results show an average colony loss rate of 46%(2022) and 17%(2023), but over 40% colony loss before wintering at 2022. Beekeepers attempt to manage their honey bee colonies in ways that optimize colony health. Disentangling the impact of management from other variables affecting colony health is complicated by the diversity of practices used and difficulties handling typically complex and incomplete observational datasets. We propose a method to 1) Varroa mite population Control by several methods , and 2) Many nursing bee put in hive before wintering.

Foot-and-mouth disease (FMD) is a highly contagious vesicular disease that affects cloven-hoofed animals, causing substantial economic losses to the livestock industry. The causative FMD virus (FMDV) comprises four structural proteins (VP1, VP2, VP3, and VP4) and several non-structural proteins. Among the capsid proteins, VP4 is the most conserved, making it an attractive target as a diagnostic and vaccine antigen, regardless of FMDV serotype. In this study, we attempted to express the VP4 protein N-terminally fused to a glutathione S-transferase (GST) tag in Escherichia coli. Whereas VP0 and VP2 proteins were expressed in the soluble fraction, we failed to detect VP4, even in the insoluble fraction. To investigate the effect of VP4 C-terminal amino acid residues on protein expression, we constructed three VP4 mutants fused to GST, among which the mutant in which the C-terminal 15 amino acid residues had been deleted showed the highest level of protein expression. Furthermore, protein expression was observed even in the mutant in which three amino acid residues (DKK) had been fused to the C terminus. However, unlike the other two mutants, the wild-type VP4 mutant was poorly expressed, thereby indicating that the C-terminal amino acid residues could play a pivotal role in determining expression of the VP4 protein in E. coli.

Royal jelly (RJ) is a gelatinous substance that bees produce to feed bees and queen bees. It’s frequently sold as a dietary supplement to treat a variety of physical ailments and chronic diseases. While it has long been used in traditional medicine, its applications in Western medicine remain controversial. The inhibitory effect of royal jelly on osteoarthritis was investigated in primary cultured rat cartilage cells and monosodium-iodoacetate (MIA)-induced arthritis rat model 10-hydroxy-2-decenoic acid (10-HAD) is the main fatty acid present in RJ. Among the criteria for RJ quality analysis, 10-HAD content has been proposed as a freshness parameter. We investigated the effect of RJ on the improvement of osteoarthritis on SD rats and they were divided into five groups. In this study, we examined the effect of enzymatic royal jelly (ERJ) administration on osteoarthritis. To determine the antiinflammatory effects of RJ, tumor necrosis factor alpha (TNF-α) and Interleukin-6 (IL-6) expression were measured after lipopolysaccharide (LPS) activation in RAW 264.7 cells. In in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. As a results, ERJ showed that TNF-α and IL-6 levels were decreased by ERJ treatment in a dosedependent manner. In conclusion, ERJ extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and cartilage cell damage. It was considered that ERJ extract may be a potential therapeutic treatment for degenerative osteoarthritis.

Ampelopsis brevipedunculata, a porcelain berry that is a kind of Korean domestic wild berry (Viticeae), has been known to be resistant to diseases and insects. A total of 2,622 unigenes containing 912 contigs and 1,710 singletons were obtained by sequencing 5,839 expressed sequence tag (EST) clones derived from the cDNA library of wild grape, A. brevipedunculata. In the gene ontology analysis, 1,175 genes related to biological process, 1,516 genes related to molecular function, and 1,413 ones related to cell components were annotated. Among the genes showing molecular function, 17 clones were classified into defense-related genes, and 102 were known to be related with responses to stress in plants. Domains, such as leucine-rich repeat, Serine/threonine protein kinase-related, WD40 repeat, EF-hand calcium-binding, pentatricopeptide repeat, and pathogenesis-related transcriptional factor were highly expressed. Genes encoding defense related proteins, such as chitinase, catalase, protein-serine/threonine kinases, were also clustered into an abundant group in cDNAs from A. brevipedunculata. Approximately, 80 simple sequence repeats with 2~5 nucleotides were detected in the cDNAs of A. brevipedunculata. These data could provide useful information for the genetic analysis of wild grapevines and in programs for breeding grape cultivars.

Transgenic chickens have been spotlighted as an highly potent bioreactor for their fecundity, short generation time, and eggs associated with mass production of protein. In this study, we generated transgenic chickens exhibiting oviduct specific expression of human growth hormone fused to human transferrin for oral administration. Gene of the modified growth hormone located at downstream ovalbumin promoter (∼3.6 kb) was introduced to stage X blastodermal cell employing retrovirus vector system. Several transgenic chickens were successfully generated. However, genomic analyses showed unexpected deletion within the transgene. The modification of the transgene seemed to occur during germ cell formation because the deletion was detected only from the sperm DNA of the G0 founder animal. There was no evidence of deletion in the somatic cell DNA samples of the same chicken. Consequently, same pattern of the deletion was confirmed in both somatic and germ cells of the G1 progeny.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is widely utilized for greenhouse pollination in South Korea. Understanding the magnitude of genetic diversity and geographic relationships is of fundamental importance for long term preservation and utilization. As a first step, we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to the “DNA barcode” region and the complete internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA from 88 individuals collected in nine South Korean localities. The complete ITS2 sequences were longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated 112-bp long repeats. The 658-bp long mtDNA sequences provided only six haplotypes with a maximum sequence divergence of 0.61% (4 bp), whereas the ITS sequences provided 84 sequence types with a maximum sequence divergence of 1.02% (21 sites). The combination of the current COI data with those of published data suggest that the B. ignitus in South Korea and China are genetically a large group, but those in Japan can be roughly separated into another group. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population were found to exist among the South Korean populations of B. ignitus, which suggests panmixia. This finding is consistent with our understanding of the dispersal capability of the species.

The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio- technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being.

The objective of this study was to evaluate in vitro production of bovine embryos in Hanwoo. Oocytes were collected by ovum pick up (OPU) from ovaries of genetically high-value Hanwoo or by needle puncture from ovaries of slaughtered cattle. OPU was done every 3 4 days duing experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, We compared the in vitro maturation rate in two groups (Experiment 1). 545 oocytes were recoverd from 4 females by 32 trials of OPU and then 433 oocytes were shown MⅡ stage after in vitro maturation (79.4%). In case of needle puncture group, 1905 oocytes were collected and then 1420 oocytes were matured to MⅡ stage during in vitro culture(74.5%). Second, we compared the developmental rate to blastocyst in two groups (Experiment 2). 1420 oocyte by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24 48 h after in vitro fertilization (IVF) was 88.6% and blastocyst development rate was 20.5% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (84.8%), blastocyst development rate was higher compared with needle puncture group (26.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio-technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being. This research was suppoted by Imsil-gun agricultural technology service center.