Human interferon alpha 2b (hIFNα-2b) is an important immune regulator widely used in clinic, for the treatment of chronic hepatitis, hairy cell leukemia, chronic myelogenous leukemia and multiple myeloma, etc. The clinically used hIFNα-2b is generally produced by E. Coli, which lacks the post-translational O-glycosylation of naturally synthesized protein, and has a short serum half-life. In this study, we report the successful generation of transgenic chickens that produce hIFNα-2b in the egg white using a feline immunodeficiency virus (FIV)-based lentiviral vector. In preliminary in vitro study, the hIFNα-2b gene under the control of CMV promoter expressed as much as 2,650 ng/㎖ in CEF-LNC-hIFNα-2bW cell. A FIV vector packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected underneath the blastoderm of freshly laid chicken eggs (stage X) to produce a hIFNα -2b transgenic chicken. Out of 187 injected eggs, 55 chicks were hatched after 21 days of incubation, and 27 of the G0 hatched chicks expressed the vector-encoded hIFNα-2b gene. The expression of recombinant hIFNα-2b in transgenic chickens constitutes an important step towards low-cost and full biological activity production of this protein drug in bioreactor. This work was supported by the Bio-industry Technology Development Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea, and by a grant from the Next-Generation BioGreen 21 Program (No. PJ011178), Rural Development Administration, Republic of Korea.

In the present study, using a MoMLV-based retrovirus vector, we successfully generated a new transgenic chicken line expressing high levels of hEPO. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). One rooster was mated to wild-type hens to produce 748 G1 progeny. PCR analysis of blood samples from these progeny revealed that there were seven G1 transgenic offspring, corresponding to a 0.9% germline transmission rate. Subsequently, Southern blot analysis of the genomic DNA from three G1 transgenic chickens was carried out to verify the stable genomic integration and copy number of the transgene in the genome. Quantitative analyses of the blood samples taken from G1 transgenic chickens resulted in 4,150 ～ 10,823 IU/㎖ (34.6 ～ 90.2 ㎍/㎖) of hEPO in the blood. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. Red blood cell numbers were more than three-fold higher in the transgenic chickens compared to the non-transgenic chickens. Successful germline transmission of the transgene was also confirmed in G2 transgenic chicks produced from crossing G1 transgenic roosters with non-transgenic hens. We confirmed that 13 transgenic chicks of 45 G2 progeny, corresponding to a 28.9% germline transmission rate. These results will help establish a useful transgenic chicken model system for studies of embryonic development and for efficient production of transgenic chickens as bioreactors. This work was supported by the Bio-industry Technology Development Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea, and by a grant from the Next-Generation BioGreen 21 Program (No. PJ011178), Rural Development Administration, Republic of Korea.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence of 5.433% (32 bp). A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35-3.97-fold lower in the USA, Canada, and Italy than that in the populations ancestral ranges.

To increase the stability of anthocyanin from black rice, β-cyclodextrin (β-CD) formed a complex with anthocyanin, and thermal and UVB stability of the complex was investigated at various pH levels. The formation of β-CD complex with anthocyanin was confirmed by the decreased UV-Vis spectrophotometer absorbance (at 511nm) with increasing the concentration of β-CD at pH 2, 2.5, 3, 4, and 5. ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid)assay was conducted to measure radical scavenging capacity (RSC) of the complex at pH 2, 4, 6, and 8, after which the RSCs were recorded over the 24 h heating(95°C) and 24 h UVB irradiating periods. The RSC values of the complexes increased as the β-CD concentration increased (0.1-2%) at pH 6 and 8. Upon thermal treatment, the RSC readings of the complexes tended to decrease to a lesser extent compared to the anthocyanin control (without β-CD) at pH 2. This result suggests that the thermal stability of anthocyanin was more retained in the presence of β-CD at low pH (pH 2). However, in the case of UVB irradiation, the effect of β-CD complex on the anthocyanin RSC was not significant, though the RSC values for both the anthocyanin control and complexes trended downward as the UVB irradiation time increased at all pH ranges except for pH 8 (no downward trend). In short, this study suggests that the effect of β-CD complex on the stability of anthocyanin antioxidant capacity depends on pH and the susceptibility to the degradation process.

Soil salinity is the most critical problem at reclaimed tidal saline soil in Korea. The objective of this study was carried out investigate the salt movement in upland soils at Gyehwa-do region in Saemangeum reclaimed land on the south Korea coasts. The experiment was conducted 5% sloped polt (3.0 m × 10.0 m) with 14 treatments and it were separated by low salinity division (LSD) and high salinity division (HSD) install. The cation content in ground water increased during time course, but in the case of land surface water the content was variable, and K+was lower than that of Na and Mg. At the LSD under rainproof condition, the salinity was directly proportional to soil water content, but at the HSD the tendency was no reversed. In condition of rainproof, the amount of soil salinity was higher at the HSD than at the LSD. Positive correlation was obtained between the soil water content and available phosphorous content at the rainfall division, but there was no significance at the surface soil of the rainproof division. Sodium adsorption ratio (SAR) and anion (Cl−, SO4 −) contents in soil were repressed in the order of vinyl-mulching > Rice straw-mulching > control plot. Our results indicate that soluble salt control is critical at reclaimed tidal saline soil and the results can provide some useful information for deciding management plans to reduce salt damage for stable crop production and diverse utilization or cultivation could be one of the management options to alleviate salt damages at reclaimed tidal saline soil in Korea.

Distillers dried grain (DDG) and makgeolli spent grain (MSG) are agricultural by-product to produce alcoholic beverage. However, they are known to contain enough nutrients. Mealworm is a promising insect resource for an animal feed ingredient as well as alternative human food. With low cost, DDG and MSG were investigated as a feed ingredient for rearing high quality mealworms. DDG and MSG were mixed with wheat bran and compared to control feed (only wheat bran) for its effects on larval survivorship, larval weight, duration for larval development, pupation rate, and pupal weight. Adding DDG on wheat bran showed positive results for larval weight, duration for larval developmental period, and pupation rate. However, adding MSG made longer duration for larval development, but it also improved larval weight, pupal weight with more than 90% pupation rate. We confirmed that adding 30～50% of DDG or MSG to conventional wheat bran have a strong potential to replace the conventional wheat bran insect feed for quality insect production.

This study was performed to apply and to utilize the digital forecasting information for Integrated Pest Management at pepper field in Chungbuk province. Chemical spray according to the forecasting prediction using FarmIPM and NCPMS were compare to the that of no control and conventional control. Pests such as Oriental tobacco budworm, Western flower thrips, and Cotton aphids were investigated the control effect by the forecasting modeling information at three pepper plots. Damaged fruit ratio (%) of Oriental tobacco budworm was ordered into no treatment (30) > forecasting (20) > conventional (12), but damages by other pest was insignificant. The frequency of forecasting control was ordered into NCPMS (31) > conventional control (17) > FarmIPM (8). Damaged fruit ratio (%) of Oriental tobacco budworm was ordered into no treatment (35.5) > NCPMS (26.1) > FarmIPM (24.6) > conventional (13.9). Density of western flower thrips was higher but had no problematic by the chemical control when it is hard to analyze the damage by cotton aphid occurrence. Damage by cotton aphids has no prediction of forecasting but done with 6th chemical spray, and their damage ratio (%) was ordered into no treatment (67.7) > FarmIPM (16.2) > NCPMS (11.3) > conventional treatment (4.8). At an analysis of economic value, gross profit was highest in FarmIPM and next to NCPMS > conventional control.

In order to understand evolutionary characteristics of gene rearrangement in Lepidoptera, we collected all available complete mitogenome (mitogenome) sequences registered in GenBank (274 mitogenomes from 44 families in 23 superfamilies as of August 6, 2015). It turned up six rearrangements that differ from the arrangement of ancestral insects, including that of the gelechioid Mesophleps albilinella that we sequenced in this study. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARNESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSEF that is found in ancestral insects. Most of the rearrangements can be explained by the tandem duplication-random loss model, but inversion, which requires recombination, is also found in two cases, including M. albilinella. Excluding the MIQ rearrangement at the A+T-rich region and ND2 junction, which is found in nearly all Ditrysia, most of the remaining rearrangements found in Lepidoptera appear to be independently derived in that they are automorphic at several taxonomic scales. Current mitogenomic data are limited, particularly for congeneric data. Thus, future research focused on congenerics could clarify evolutionary independency at the generic level also.

The Gelechioidea is the second most species-rich group of Lepidoptera, but only limited number of mitochondrial genome (mitogenome) sequences is available. Thus, we sequenced the complete mitogenome of a gelechioid Hieromantis kurokoi (Lepidoptera: Stathmopodidae) to use the data for future study for the higher phylogeny of Ditrysia in Lepidoptera. The arrangement of the genome was identical to typical one found in Ditrysia (trnM-trnI-trnQ) (underline for inverted gene). The COI began with CGA, which has been designated as the start codon for majority of lepidopteran species, whereas other protein-coding genes (PCGs) began with the typical ATN codon. The 360-bp long A+T-rich region harbored the conserved sequence blocks Phylogenetic analysis using the 13 PCGs both by Bayesian inference (BI) and Maximum-likelihood (ML) methods indicated that H. kurokoi belonging to the family Stathmopodidae grouped together with within-familial species Atrijuglans hetaohei with the highest nodal support (BI, 1.0; ML, 100%).

Soil salinity at newly reclaimed land in west coast of Korea is highly variable and depending on soil characteristics and weather conditions. The objective of this study was to evaluate the spatiotemporal distribution characteristics changes on soil electrical conductivity and exchangeable sodium percentage in Saemangeum newly reclaimed land. Experimental sites was selected at Gyehwa (35o75'N, 126o60'E) in Saemangeum reclaimed land and their dominant soil series was Munpo (coarseloamy, mixed, non-acid, mesic, typic Fluvaquents). Soil samples were periodically collected at 0 ~ 20 cm and 20 ~ 40 cm layer from each site. Soil electrical conductivity had a wide range from 0.15 to 41.76 dS/m, which was variable according to the weather conditions. The average soil electrical conductivity from March 2014 to 2015 was 6.4 and 3.4 dS/m at Gyehwa in Seamangeum reclaimed land. Calcium concentration in soil solution was negatively correlated with soil electrical conductivity and soluble sodium. Soluble sodium concentration had great variations and it was the most influencing single factor for temporal variations of soil electrical conductivity regardless of soil textural properties. The characteristics of Saemangeum reclaimed land had different shares of saline and sodic properties during the experimental period. Our results indicate that persistent monitoring and modeling on soil salinity at coastal tide land is fundamental and the results can provide some useful information for deciding management plans for diverse utilization or to reduce salt damage for stable crop production at reclaimed tidal land.

The present study prepared molybdenum trioxide (MoO3), the most important intermediate of molybdenum metal, by using a fluidized bed reactor for the thermal decomposition of ammonium molybdate (AM) in the presence of an air flow. During the process of fluidizing the sample inside the reactor, the reaction time and temperature were optimized with a close analysis of the X-ray diffraction (XRD) data and with thermogravimetric analysis (TGA). In particular, the temperature level, at which the AM decomposition is completed, is very important as a primary operating parameter. The analysis of the XRD and TGA data showed that the AM decomposition is almost completed at ~350 oC with a reaction time of 30 min. A shorter reaction time of 10 min. required a higher reaction temperature of ~500 oC with the same air flow rate to complete the AM decomposition. A sharp rise in the decomposition efficiency at a temperature ranging between 320 and 350 oC indicated a threshold for the AM decomposition. The operating conditions determined in this study can be used for future scale-ups of the process.

Grapholita molesta occur four times a year and Carposina sasakii occur twice a year, and both pests do damage on stone fruits such as peach, apple, plum, apricot, etc. Grapholita molesta is worldwide distributed in temperate and subtropical areas including South Korea. But, Carposina sasakii distributed in South Korea, Japan, China and Asia, and has been managed as an important import quarantine pest by the authorities of United States, Canada and Taiwan. Forecasting of both pests in Korea is currently done through the investigation of 1,000 fruits per 10 trees (100 fruits / tree) in designated peach orchard. However, this method is very difficult to observe the pest by bagging of peach and require too much time and labor. Therefore, we tried to carry out a new forecasting method by using of sex pheromone traps for newly standardized method as an alternative. Using sex pheromone trap, attractiveness of G. molesta was proved to be 2.5 > 1.5 > 0.5 m by the height and the border => outside > center by the position. Attractiveness of C. sasakii made no difference in height, but, more trapped at the center and border than outside in position at peach orchard.