The entomopathogneic fungal colonization has become an interesting issue in ecological pest management. Plant-orientedfungi induce plant growth promotion and act as natural control agents against insect pest. Herein this work, we evaluatedthe ability of fungal colonization on bark, branches and leaves of pine tree (Pinus densiflora) to select optimal candidatesfor controlling Japanese pine sawyer (Monochamus alternatus), which is the pine wilt disease vector in Korea. As a result,some Beauveria and Metarhizium isolates could colonize on the substrates of P. densiflora and showed pathogenicity againstM. alternatus. This result suggests that the selected entomopathogenic fungi are possibly candidates for long-term managementof M. alternatus in pine tree forest.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopeniasyndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest.We evaluated three bioassay methods, such as spray, dipping and dropping. As a result, the dipping method was themost efficient way to measure the fungal virulence. Finally, we constructed a Haemaphysalis longicornis pathogenic fungallibrary to further facilitate the resources to be used as potential biological control agents. A conidial suspension of 1×107conidia/ml was exposed to the ticks by dipping. We confirmed that some isolates were highly virulent to the ticks. Thisresults indicated that some fungi could be used to effectively control the Haemaphysalis longicornis.

This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

The mesenchymal stem cells (MSC) has been investigated as a source of stem cell therapy to replace and treat damaged cells. Human endometrial epithelial and stromal cells was isolated from hysterectomy tissue and the direct evidence of stem/progenitor cells in the human endometrium was identified. Endometrium derived stem cells (EnMSCs) are known to have a high proliferative ability, genetic stability, lack of tumorigenicity and low immnunogenicity during long-term cultivation. Here, we aimed to identify MSC in canine endometrium and characterize its potential to differentiate into decidua cells. EnMSCs were isolated from thrown-away spayed uterus of adult canine depending on their estrus cycle, and identified by flow cytometry, immunocytochemistry and flow cytometry with MSC specific markers. We then characterized the ability of EnMSCs by the doubling-time analysis, colony-forming units and MSC differentiation assays. Isolated EnMSCs expressed stem cell specific genes (Sox2, Oct4, Nanog, MCAM, Endoglin, Susd2 and IGTB) and MSC surface markers (CD90, CD44 and CD117). EnMSCs are also differentiated into adipogenic, osteogenic and chondrogenic cells morphologically under modified conditions with the expression of lineage specific genetic markers. EnMSCs showed higher proliferation ability than canine amniotic fluid derived MSCs which were used as a positive control. EnMSCs were cultured at low density (10, 20, cells/cm2) and initiated to form small colonies of loosely-arranged cells and gradually formed large colonies of densely-packed cells which underwent self-renewal with high proliferative potential which is similar to the clonogenicity feature of human endometrium-derived stem cells. EnMSCs were then induced to differentiate into decidua cells with 0.5 mM dbcAMP. After 14 days, EnMSCs changed their morphology into the elongated and rounded shape. The induced decidual cells expressed PRL and IGFBP1 which are typically expressed in decidua cells. In conclusion, we successfully isolated and characterized MSC in the canine endometrium which differentiated into decidua cells. These results showed that endometrium may be a promising source of stem cells, and furthermore raise the possibility of canine EnMSCs as a novel hypothetical decidualisation model of infertility associated with decidualisation insufficiency and implantation failure.

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.

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 present study was made to assess annual changes on seasonal occurrence of Apiformes, commonly known as bees, as global warming apparently happens. Bees were collected from ten sites using a malaise trap for each site, and sample collection was made every two weeks from April to September in 2011 to 2014. In overall diversity, abundance and richness of bees appeared higher in northern sites, particularly Gwangneung and Hongcheon, than southern. The abundance of bees declined sharply in 2013 and decreased even further in 2014. Among the bee families the most decreased families were Halictidae and Andrenidae. In order to see the effect of annual temperature fluctuation correlation analysis was used with collection data and weather data. However, the result indicated that the decline of the bees was not significantly affected by changes of temperature although most of the bee families except Halictidae showed negative relation with temperature data(p < 0.05). In addition we also analysed the bee collection data and flowering data during spring in Gwangneung. Dufrene-Legendre indicator species analysis (ISA) for latitudinal sites identified seventeen indicator species (p < 0.05) that were limited to northern region.

In the present study, Cydia kamijoi Oku is newly recognized in korean insect fauna. This tortricid moth was first found damaging the cones of Abies koreana in Jeju Island 2014. The moth can be a serious insect pest on A. koreana because of high damage rate on the cones, up to 71% average. The genus Cydia now was 11 korean species including C. kamijoi. Regarding this species, some basic information such as collection records, morphological characters, and ecology were provided

This study was carried out to investigate insect community structure from different habitats. We performed day and night collection at three different habitats (mountain, coast and rural area) of island Deokjeok, island Soya and island Mungap from May to September in 2014. A total of 3,482 individuals of 725 species, 119 families belonging to 10 orders were collected and identified. A dominant species was Corymbia rubra (Cerambycidae) despite a very low percentage of the species among the catches. Results of ANOVA test showed a significant effect of habitats typeon species diversity. Also, combination of seasons and habitats types were significantly influential with species abundance and species diversity. Indicator species analysis (ISA) result identified 121 significant (p < 0.05) indicator species; one species for the habitats cluster, 93 species for the season cluster and 27species for combination of habitats with seasons.

An insect faunal survey was carried out to investigate insect community structure along the vegetation community to monitor insect species in forest ecosystem. We performed day and night collections from June to August along three vegetation communities of Is. Nam-hae in 2014: the first stand with Pinus thunbergii, the second P. thunbergii with Quercus serrata and the third P. thunbergii with various Quercus species. In total 2,259 individuals of 532 species, 99 families, 13 orders are identified. Cluster Analyses (CA) showed that all three vegetation communities were relatively similar between vegetation community types. According to indicator species analysis (ISA) result, nine significant indicator species were identified (p < 0.05); five species were found to be affected by the vegetation cluster and four species the month cluster.

To date there have been only two species in genus Pogonus and subgenus Europhilus of genus Agonum recognized from Korean insect fauna: Pogonus itoshimaensis Habu, 1954 and Agonum (Europhilus) bellicum Lutshnik, 1934. In the present study one additional species for each genus is newly recognized from Korea: Pogonus (Pogonus) japonicus Putzeys, 1875 and Agonum (Europhilus) gratiosum nipponicum Habu, 1972. A key to adults, redescription, diagnostic photos of adult and male and female genitalia are provided.

Insect pollinators of the endanger orchid Cypripedium japonicum were surveyed and identified during two years, as a part of a conservation project of the orchid at Jukyeup-san and Hwaak-san (Mt.), South Korea. In total 40 individuals of 16 species in 4 families were identified. The dominant family was Halictidae, and Lasioglossum exiliceps Vachal visited the most frequently C. japonicum during the surveys. The average visiting frequency was 2.5 individuals per hour and the highest 4.3, from 12:00 – 13:00 in a day. After 15:00 insects did not visit the flowers at all. However, all of the visiting insects were found to not carry a pollinium or pollens of the orchard on their bodies; pollen carryover by any of the visiting insects did not occur at all. The orchid seems to require certain pollinators in particular body thickness due to its unique pollination mechanism. The orchid has two exit route openings, around 1 cm in diametre, where the entrapped insects can exit and an anther is situated just in front of each opening. It was inferred that a pollen carrier should be around 1 cm in body thickness. Therefore, the candidate species as the proper pollen carriers can be Tetralonia nipponensis Perez, Xylocopa appendiculata circumvolans Smith and Bombus consobrinus Dahlbom among the surveyed visitors.

본 실험은 토마토 플러그묘의 접목활착율과 묘소질 향상을 위한 적정 상토 내 수분함량을 구명하기 위하여 수행되었다. 시험구는 접수와 대목의 상토내 수분함량을 각각 3처리구(고, 중 저)씩, 총 9개의 조합을 설계하여 각 처리구별로 접목활착율과 묘의 생육을 조사하였다. 접수의 상토내 수분 함량의 차이는 접목 활착율에 통계적 유의차는 나타나지 않았으나, 대목의 수분함량은 접목활착율에 영향을 미쳐 수분함량이 낮아질수록 활착율도 저하하는 경향을 보였다. 수분 함량에 따른 묘소질도 접목활착율과 유사한 경향을 보여 접수의 수분함량차이는 생육지표에 큰 영향을 미치지 않았으나, 대목에서는 잎(엽수, 엽장, 엽폭)을 제외한 초장, SPAD함량, 경경은 수분함량의 차이에 따라 유의적 차이를 나타냈다. 묘의 충실도는 상토내 수분 함량이 접수는 중간, 대목은 높은 조합에서 가장 좋게 나타났다. 근권부의 생육에서도 대목의 수분함량이 주로 영향을 미쳐, 수분함량이 저하될수록 전 뿌리 표면적, 전 근장, root tip수 모두가 감소되는 것으로 나타났다.

A long term monitoring was carried out to compare altitudinal effects on insect community structures on high mountains and evaluate responses the monitoring and changed of insect communities induced by the climate change. These mountains were choose for Mt. Jeombong for northern part, Mt. Ilwol for middle and Mt. Beakun for southern. Each mountain was divided into three altitudinal gradients. They were collected three times a season from spring to autumn, using pitfall traps for ground beetles and UV light trap for moths. The present study presents preliminary results of analysis for the first year monitoring. In total 41beetle species and 326 moths were collected from the monitoring sites in 2012. abundance of ground beetles and moths were the highest in Mt. Baekun followed by Mt. Ilwol and lowest in Mt. Jeombong. Analysis of variance (ANOVA) results showed statistically significant differences among sampling area, species evenness and Shannon’s diversity index with altitude in species abundance as a response variable. Also we found statistically significant differences to three species of ground beetle and six species of moths with altitude. Although we expected a distinct cluster with the difference of altitude at each study site. one of ordination analysis, nonmetric multidimensional scaling (NMS), showed distinct clusters with the ground beetles assemblage at some altitude and moths assemblage at sampling date.

We report a procedure for direct shoot regeneration via leaf segments of Lilium Oriental hybrid ‘Casa Blanca’. The segments were cultured with the abaxial side in contact with a Murashige and Skoog medium containing 0.1 mg • L-1 BA and 0.1 mg • L-1 NAA, 3% sucrose, and 0.8% agar for shoot regeneration. The cultures were incubated for 4 weeks under a 16 hrs photoperiod at 23 ± 2oC for adventitious shoot regeneration. With this procedure, a mean shoot regeneration frequency of 88-90% and a mean number of shoots of 2.5-3.6 per segment were obtained. Ploidy analysis of the regenerated plants using flow cytometer revealed the same ploidy level (diploid) with mother plant. This study will be used for large-scale multiplication and genetic transformation system in lily.

Escape pattern of the ambrosia beetle Platypus koryoensis (Murayama) based on individual gallery was investigated. The survey was conducted in four oak forests located in Chuncheon, Hanam, Paju and Seongnam, and escape patterns from individual gallery were surveyed using emergence trap attached at the entrances of galleries formed in 2010. The number of adults caught in each trap was periodically counted from May 2011 to September 2012. The adults ambrosia beetle was caught over two years after infestation. The escape patterns in the first and second year after infestation showed different types; in the first year, escape of adults from gallery was observed one or two times whereas escape from gallery was observed two times in the second year with variation in aspect of escape density and timing. Moreover, variations in escape pattern between trees or between galleries in the same tree was observed. Colonization success rate in living tree was higher than that of dead tree, but the number of progeny per gallery in living tree was lower than that of dead tree. Colonization success rate in the galleries of second year was lower than that of first year, but the number of progeny per gallery of second year was higher than that of first year.