In March of 2015, severe sooty mold was observed on blueberries cultivated in a farmer’s greenhouses in Goseong-gun, Gyeongsangnam-do, South Korea. Typical symptoms included a black or dark brown, superficial fungal growth on the leaves, stems, and fruit of blueberry plants. The fungal colonies on PDA were flat and black. The conidia were dark green in color, globose to sub-globose in shape and 3-5×3-4 μm in size. The ramoconidia were cylindrical and 18-42×3 μm in size. The conidiophores were dark brown, cylindrical-oblong in shape and 42-138×3-4 μm in size. To confirm the identity of the fungus(F130), the 18S rDNA as well as actin and β-tubulin gene sequences were PCR-amplified and sequenced. The pathogenicity of the isolate was then explored on 2-month-old potted blueberry plants. Based on the mycological characteristics, molecular identification, and pathogenicity on host plants, this fungus was identified as Cladosporium sphaerospermum. This is the first report on sooty mold of blueberry caused by C. sphaerospermum in Korea.

The ovum pick up(OPU) technique can be used to produce embryos after in vitro culture of ovarian oocytes, can be used for early securement for effective herd early proliferation and excellent Hanwoo genetic resources, It is attracting attention as a very important technique for establishing technology. In addition to in vitro culture techniques, the number of oocytes retrieved depends on the environment and timing of the OPU. This study was conducted to compare and examine seasonal effect to the differences in the number of recovered oocytes, recovery rate and embryo development rate using Korean cattle kept in animal genetic resource research center by OPU technique. The grade of COCs was evaluated by microscopic examination. Grade A had 3 or more layers of cumulus cell and compact cytoplasm. Grade B had 1~3 layers of cumulus cell and compact cytoplasm. Grade C had 1 layers cumulus cell and compact cytoplasm. Grade D was denuded oocyte and poor cytoplasm. The recovery rate was 47.8±3.4% in summer (June to August) and 51.6±3.8% in autumn (September to October). The number of oocytes was 5.7±0.6 in summer and 5.2±0.7 in autumn. Oocyte grade A and B was 46.2%±6.3% in summer and 51.1±5.0% in autumn. The cleavage rate was 46.1±7.1% in summer and 65.0±11.3% in autumn. Blastocyst development rate was 19.9±9.4% in summer and 29.0±8.7% in autumn. There was no difference the recovery rate of oocytes and the number of embryos between summer and autumn. Cleavage rate and blastocyst rate of autumn was higher than summer. we will investigate to study the appropriate method for the production of Hanwoo embryos and the systematic comparison.

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Recently, polymorphisms reported to be significant association with sperm MOT. This study was conducted to evaluate the SNP in the coding region of ESR1 (g.672C>T inexon 1) as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results,The g.672C>T was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP Also, the SNP was low significantly associated with ALH.Therefore, we suggest that theSNP in the coding region of ESR1 (g.672C>T in exon 1) may be used as a molecular marker for Duroc boar Post-thawed semen quality.

Estrogen receptor 2 (ESR2) is involved in oestrogen-related apoptosis in cell cycle spermatogenesis but their effects have not yet confirmed in pig. Therefore, this study was performed to investigatetheir association with semen motility and kinematic characteristics. DNA samples from 126 Duroc pigs with records of sperm motility and kinematic characteristics [Total motile spermatozoa (MOT), Curvilinear velocity(VCL), Straight-line velocity(VSL), the ratio between VSL and VCL(LIN), Amplitude of Lateral Head displacement(ALH)] were used in present study. A single nucleotide polymorphism (g.35547A>G) was associated with MOT, VCL, VAP and ALH in Duroc population (p < 0.05). Therefore, we suggest that the porcine ESR2 may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not clear yet. These results will improve the understanding of the functions of the ESR2 in spermatogenesis within the reproductive tracts, and will shed light on ESR2 as a candidate gene in the selection of good sperm quality boars.

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

Cryopreservation of bovine embryos is used to efficiently implant surrogate mothers. It has been widely accepted that high lipid content in the oocyte interrupts its survival during freeze-thaw cycles. Serum component in the culture medium is thought to increase the embryo`s lipid contents. Conversely, L-carnitine stimulates lipid metabolism by transporting long chain fatty acids into the mitochondria. Objective of this study was to analyze the effect of L-carnitine supplementation in IVM medium and defined IVC medium on the development, lipid contents and the cryosurvival of bovine IVF embryos. 0.0, 1.5, 3.0 and 6.0 mM L-carnitine was supplemented in IVM medium, respectively (IVM-LC 0.0, LC 1.5, LC 3.0 and LC 6.0). Development rate from the 2cell to the morula stages was higher in IVM-LC 3.0 groups than those of IVM-LC 6.0 (p<0.05). But there were no significant differences among the other groups in the blastocyst rates and lipid content results. When 0.0, 1.5, 3.0 and 6.0 mM L-carnitine were supplemented in IVC medium (IVC-LC 0.0, LC 1.5, LC 3.0 and LC 6.0), development competence was not significantly different between those embryos. Lipid contents of embryos treated L-carnitine (IVC-LC 1.5, 3.0 and 6.0) were significantly lower than embryos of non-treated group. L-carnitine was supplemented 0.0, 1.5, 3.0, 6.0 mM during IVM and 3.0 mM during IVC (LC 0.0 - 3.0, LC 1.5 – 3.0, LC 3.0 – 3.0, LC 6.0 – 3.0) and cryosurvival of blastocysts confirmed after freezing-thawing. There were no significant differences on development, but LC 3.0 – 3.0 was significantly lower lipid contents than other groups. And LC 3.0 – 3.0 had better survival rates and hatched rates of blastocysts than LC 0.0 – 0.0. In conclusion, supplementation of L-carnitine in defined IVC medium decreases lipid contents. And L-carnitine supplementation improves cryosurvival and developmental ability of bovine IVF embryos.

To improve survival rates of vitrified pig oocytes, the treatment of cytoskeletal stabilizer on an appropriate time is one of the possible approaches. However, the exact treatment timing and effect of cytoskeletal stabilizer such as cytochalasin B (CB) is not well known during oocyte vitrification procedures. Thus, the present study was conducted to determine optimal treatment timing of CB during vitrification and warming procedures. In experiment 1, the survival rates of the post-warming pig oocytes were analyzed by fluorescein diacetate (FDA) assay with 4 classifications. In results, post-warming oocytes showed significantly (p<0.05) decreased number of alive oocytes (31.8% vs. 86.4%) compared to fresh control. In detail, the significant difference (p<0.05) was found only in strong fluorescence (18.2% vs. 70.5%) not in intermediate fluorescence groups (13.6% vs. 15.9%). In experiment 2, CB was treated before (CB-Vitri) and after (Vitri-CB) vitrification. In results, group of Vitri-CB showed significantly (p<0.05) higher (91.6%) survival rates compared to group of CB-Vitri (83.7%), significantly (p<0.05) and comparable with group of Vitri Control (88.7%) by morphological inspection. In FDA assay results, group of Vitri-CB showed significantly (p<0.05) higher (44.2%) survival rates compared to groups of CB-Vitri (36.7%) and Vitri Control (35.1%). In conclusion, the increased survival rates of post-warming pig oocyte treated with Vitri-CB method are firstly described here. The main finding of present study is that the CB treatment during recovery could be helpful to refresh the post-warming pig oocyte resulting its improved survival rates.

The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.

쌀가루의 활용도를 높이기 위해서 제과 품목 중에 쌀파이를 제조하였고, 쌀파이에 볶음땅콩가루, 새싹땅콩가루를 첨가하여 쌀파이의 품질 특성을 분석하였다. 수분함량은 밀 파이에 비해 쌀파이가 낮았고, 볶음땅콩가루, 새싹 땅콩가루 등 부재료를 첨가했을 때 수분함량이 낮게 나타났다. pH는 밀파이에 비해 쌀로 만든 파이에서 pH가 낮게 나타났으며, 볶음땅콩가루나 새싹 땅콩가루를 첨가하였을 때 pH가 높게 나타났다. 반죽의 밀도는 밀파이에 비해 쌀로 만든 파이에서 밀도가 낮게 나타났으며, 볶음땅콩가루나 새싹 땅콩가루를 첨가했을 때 쌀파이보다는 밀도가 높게 나타났다. 손실률은 밀파이에 비해 쌀로 만든 파이에서 높게 나타났으며, 볶음땅콩가루나 새싹 땅콩가루를 첨가했을 때 쌀파이보다는 손실률이 낮게 나타났다. 쌀로 제조한 파이에서 볶음땅콩가루와 새싹땅콩가루를 첨가했을 때 명도(L)와 황색도(b)는 감소하고, 적색도(a)는 증가한 경향을 보였다. 쌀파이의 경도는 밀파이보다는 쌀가루로 만든 파이에서 경도가 증가하였으며, 볶음땅콩가루, 새싹땅콩가루를 첨가할수록 경도는 낮아졌다. 쌀파이의 지방산 조성은 포화 지방산은 myristic acid, palmitic acid, stearic acid로 나타났다. 단일불포화지방산은 주로 oleic acid로 구성된 것으로 확인되었고, 다가불포화지방산의 조성은 linoleic acid로 구성된 것으로 확인되었으며, 밀에 비해서 쌀가루로 제조한 파이에서 높게 나타났다.

Epizootic HPAIV, H5N6, and H5N8 infections produced severe loss in poultry and wild birds in the Republic of Korea from 2016 to 2017. But pathological lesions and antigen distribution of the novel HPAIV H5N6 clade 2.3.4.4 in natural cases have been rarely reported. Herein, we describe the pathological lesions and antigen localization in chickens (layer and Korean native), ducks, and Japanese quail naturally infected by HPAIV H5N6. Grossly, severe reddening, swelling, and some necrotic foci, which were similar to septicemia or viremia, were observed in skin and many visceral organs including trachea, lung, liver, spleen, and pancreas. Histopathologically, pulmonary congestion and edema, as well as necrotizing hepatitis, splenitis, pancreatitis, myocarditis, and encephalitis were observed. Immunohistochemically, numerous HPAIV antigens were detected in necrotic parenchymal cells and in blood vessels of the respiratory, lymphoid, digestive, urinary, nervous, and cardiovascular systems. The results indicate that HPAIV H5N6 spread to the entire body via blood and caused severe damage throughout the entire body. The HPAI H5N6 clade 2.3.4.4 virus was isolated from samples of all four cases.