Somatic cell nuclear transfer (SCNT) is the technique which generates embryos by transferring diploid nucleus into an enucleated oocyte, it has produced specific animals successfully in a variety of species. However, the developmental capacity of SCNT embryos is still relatively lower than that of embryos produced in vivo. Oocyte is a kind of lipid rich cells, its quality limits the efficiency of embryo production. L-carnitine is a co-enzyme facilitating the transportation of long chain fatty acids across the inner mitochondria membrane where fatty acids are used for generating adenosine triphosphate (ATP) via beta-oxidation. It also has antioxidant actions which may protect mitochondrial membranes and DNA against damage induced by reactive oxygen species (ROS). Whether L-carnitine is functional in bovine SCNT embryos are unknown. Therefore, the objective of this study was to examine the effects of L-carnitine on oocyte maturation and developmental competence of subsequent SCNT embryos. L-carnitine was supplemented during IVM, then intracellular ROS and GSH levels, mitochondrial activity, gene expression of COCs were analyzed at the end of IVM. SCNT embryos were produced subsequently, apoptosis detection and gene expression evaluation were performed in blastocysts. In the results, treatments with 1.5 mM and 3 mM L-carnitine significantly improved maturation rates (P<0.05). Treatments with 3 mM L-carnitine effectively induced improvement in nuclear maturation, intracellular GSH levels and mitochondrial activity, as well as a reduction in intracellular ROS levels (P<0.05). mRNA levels of CPT1A, ACAA1, ACAA2, AREG, EREG, SOD1, GPX4, GLUT1 and CDC2 transcripts were effectively up-regulated by 3 mM L-carnitine treatments (P < 0.05). Similarly, 3mM L-carnitine induced an increase in blastocyst developmental rates and an improvement in blastocyst quality (P<0.05). Our study indicates that L-carnitine treatment during IVM improves oocyte nuclear maturation and subsequent SCNT embryo development.

Bovine somatic cell nuclear transfer (bSCNT) embryos can develop to the blastocyst stage at a rate similar to that of embryos produced by in vitro fertilization (IVF). However, the efficiency of somatic cell cloning has remained low, and applications have been limited, irrespective of the nuclear donor species or cell types. One possible explanation is that the reprogramming factors of each oocyte is insufficient or not properly adapted for the receipt of a somatic cell nucleus, because it is naturally prepared only for the receipt of a gamete. Here, we would like to introduce the aggregation method (agSCNT), a new experimental system that enables and increase oocyte volume and examined its subsequent development. Judgement by the blastocyst formation rate or total cell number was significantly higher in the agSCNT group than that in the SCNT group, and was very similar to that in the control IVF group. Moreover, the cleavage formation rate in the agSCNT group (61.5 ± 1.3) was higher than that in the SCNT group (39.7 ± 2.1), while still less than that in the IVF group (75.4 ± 1.3). We also analyzed the epigenetic modifications in bovine IVF, agSCNT, and untreated SCNT embryos. In conclusion, the present study demonstrated that agSCNT improves the in vitro developmental competence and quality of cloned embryos, as evidenced by increased total cell numbers (TC).

Embryo transfer is one of the important process of assisted reproductive technology (ART) and that is associated with uterus endometrial receptivity. Recently, mouse endometrial stimulation by artificial injury had shown the favorable effect on conception. In this experiment, we used uterus stimulation method that injury the endometrium to increase implantation rate for spontaneous Diabetes Mellitus (sDM) rat. Rats are divided into several groups involved a control group. We performed the surgical method to Experimental group bilaterally or unilaterally After that, we investigated morphological change and calculated implanted embryos respective sides of the uterus. The number of implanted embryo in the experimental group was significantly higher and there were lots of morphological changes including glands and endometrial cells that support implantation. Our results showed that rat uterus endometrial injury in ART help enhancing implantation rate.

한우 사육은 소비자의 요구와 동시에 농가의 소득을 향상시키기 위하여 고급육 생산을 목표로 사육되고 있다. 하지만 오랜 기간의 사양기술의 발전에도 불구하고 아직까지 거세비육을 하는 농 가에서 요로결석 발생으로 인한 피해가 종종 발생하고 있다. 요로결석은 비육후기 복통 및 배뇨장 애에 의한 스트레스로 인한 증체량 감소뿐만 아니라 심한 경우에는 폐사에 이르기도 한다. 본 연 구는 국립축산과학원에서 동일한 연령과 사양 프로그램으로 사육되던 거세비육우 22개월령 24두 중 1두에서 이상증상이 발생하여 수의사가 현장에서 진단을 하였다. 해당 가축은 전형적인 요로결 석 증상을 보였으며 배뇨를 못하는 요도폐색 증상을 나타내어 긴급도축을 하였다. 그러나 긴급도 축 2주 후 나머지 23두 중 1두에서 동일한 증상이 추가로 발생하여 긴급도축을 하였으며 수의사 가 도축장에 동행하여 방광과 요도내에 결석유무를 확인하여보았다. 해당 긴급도축우의 방광을 확 인하여 본 결과 방광근육 출혈 및 방광내에 결석존재와 결석으로 인한 요도폐색을 직접 확인하였 다. 2두의 긴급도축 후 남은 22두의 거세 비육우에서는 30개월령 출하 시기까지 더 이상의 요로결 석 임상증상이 발생하지 않았다. 하지만 동일 연령과 사양 프로그램으로 비육된 축군에서 방광 내 결석이 존재할 수 있다는 가정하에 나머지 22두의 출하시에 도축장에 동행하여 방광 내 결석유무 를 확인하였다 22두 중 개체에 따라서 결석 크기와 형태 및 양에서 차이가 있었지만 11두(50%)에 서 방광 내 결석을 확인하였다. 결석 성분분석결과 Carbonate apatite 20%, Magnesium Ammonium Phosphate (struvite) 80% 성분으로 이루어진 결석과 Calcium Oxalate Monohydrate 50%, Uric acid 50% 성분으로 구성된 2종의 결석을 확인할 수 있었다. 또한 13두에서는 방광내에 혈액, 방광조직 출혈, 점막비후 등 육안적으로 이상소견을 확인 후 조직학적 검사결과 염증소견을 확인할 수 있었 다. 방광 내 결석이 발견된 11두 전 두수에서 방광염증이 확인되었으며, 방광내에 결석이 발견되 지 않은 11두 중 2두에서 방광염증이 확인되었다. 방광 내 결석존재와 방광염증 발생에 있어서 유 의성을 확인할 수 있었다.

본 연구는 농후사료 다량 급여 시 나타나는 산통은 발생하지 않으면서 증체량은 높일 수 있는 적정 농후사료 급여 횟수를 설정하기 위해 수행하였다. 제주산마 13두를 이용하여 12주간 농후사 료 급여 횟수(체중의 2.5%를 2, 3, 4회)를 달리하여 급여하였다. 이때 건초와 물은 무제한 급여하 였다. 번식마의 농후사료 급여 횟수에 따른 산통 발생을 조사한 결과 혈액 내 피브리노겐 함량은 처리구간 유의적 차이가 없었다(p>0.05). 농후사료 급여 횟수별 증체량 조사 결과, 총 증체량은 일 일 4회 급여구에서 높아지는 경향을 보였고, 일당증체량은 8~12주에서 일일 4회 급여구의 경우 높 아지는 경향을 보였으나 유의적인 차이는 없었다(p>0.05). 장내 pH와 직장온도는 모든 처리구에서 정상 범위 내에 있었고, 유의적인 차이가 없었다(p>0.05). 이상의 결과를 종합하면 번식마에게 체 중의 2.5%의 농후사료의 급여 횟수 조절에 따른 산통 발생과 증체에는 차이가 없는 것 판단된다.

This study determined diagnostic threshold of polymorphonuclear cells (PMN) for cytological endometritis (CEM) and its impacts on reproductive performance in dairy cows. Uterine cytology using 407 Holstein cows was performed at 4 weeks postpartum and proportion of endometrial cells and PMN was evaluated. A receiver operating characteristics curve was built to assess the cutoff level above which the PMN proportion affected the hazard of pregnancy by 200 days postpartum. The cutoff level for CEM was set at ≥14% PMN (sensitivity = 31.3%, specificity = 81.7%, P < 0.05). Cows with CEM had a lower probability of resumption of cyclicity (OR = 0.58 P < 0.05) and lower hazard of pregnancy by 200 days postpartum (hazard ratio = 0.58, P = 0.0001) than cows without CEM. Cows with CEM tended to have a lower probability of pregnancy to the first insemination (OR = 0.65, P < 0.1) and a greater number of insemination per conception (2.3 vs 2.2, P < 0.1) than cows without CEM. In conclusion, the threshold level of PMN was 14% to define CEM at 4 weeks postpartum and CEM decreased subsequent reproductive performance in dairy cow

Post-ejaculation of sperms into the female reproductive tract, acquisition of sperm capacitation is an essential step in the fertilization process. Accordingly, during in-vitro fertilization, the successful fertilization requires necessarily induction of capacitation in the retrieved sperms. To date, many candidate substances have been considered as capacitation inducers. However, there were no reports about the comparison of efficiency inducing sperm capacitation among diverse capacitation inducers. Therefore, we tried to determine an inducer showing the best capacitation performance in mouse sperms by comparing the preimplantation development of in-vitro-fertilized embryos using sperms experiencing capacitation by a variety of capacitation inducers. For these, calcium, progesterone, bovine serum albumin (BSA), heparin, lysophosphaticylcholine (Lyso-PC) were used as candidate capacitation inducers. Optimized concentration of each inducer were determined by accessing ratio of sperms experiencing acrosome reaction using coomassie G-250 blue staining. Subsequently, in vitro fertilization was performed using sperms incubated in each optimized concentration inducer. The ratio of fertilized oocytes was observed. As the results, Calcium at 2.7 mM and 0.3% (w/v) BSA showed the highest fertilization rates compared to 15 μM progesterone, 50 mM heparin, and 100 μM Lyso-PC. From these results, we found that 2.7 mM calcium and 0.3% (w/v) BSA were the most effective sperm capacitation inducers of mouse sperm for in vitro fertilization. From these results, we could identify that, among diverse sperm capacitation inducers, 2.7mM calcium and 0.3% (w/v) BSA were the most effective inducers for in vitro fertilization.

Generally, fate of spematogonial stem cells (SSCs) can be determined specifically by microenvironments enclosed with various extracellular matrix (ECM) components and integrins recognizing directly ECM proteins play an pivotal role in transporting ECM-derived signals into cytoplasm, resulting in inducing a variety of biological functions such as cell attachment, self-renewal and differentiation. However, to date, studies on type of integrins expressed on the undifferentiated SSCs remain unclear. Therefore, we tried to investigate systematically what kind of integrin subunits are expressed transcriptionally or translationally in the SSCs derived from testis of hybrid B6CBAF1 mouse. For these, isolation of SSCs from testis were conducted by magnetic activated cell sorting (MACS) using Thy1 antibody. Subsequently, transcriptional and translational level of integrin α and β subunits in the isolated SSCs were measured by real-time PCR and fluorescene immunoassay, respectively. As the results, transcriptional levels of genes encoding total 25 integrin subunits were quantified, and integrin α4, α6, α7, α9, αV, αL and αE and integrin β1, β5 showed higher expression levels than other subunits. By contrast, integrin α3, α5, α 10 and α11 and integrin β2, β3, β4, β7 were weakly transcribed. When translational levels of the integrin α subunits showing high transcription level (α4, α6, α7, α9, αV αL, and αE) were measured, integrin α6, α7, α9, αV and αL were higher than integrin α4 and αE. In case of integrin β subunit, β1 evaluated more expression than β5. From these results, we speculate that the undifferentiated SSCs derived from hybrid B6CBAF1 mouse may express integrin α4β1, α6β1, α7β1, α9β1, αVβ1 and/or αVβ5 on plasma membrane. Moreover, this information will greatly contribute to constructing non-cellular niche supporting self-renewal of SSCs in the future.

The present study was conducted to investigate the effect of BHT supplementation on sperm motility, viability, acrosomal integrity and plasma membrane integrity after frozen-thawing. One ejaculate was collected from one fertile Hanwoo bull by using artificial vagina at Hanwoo Research Institute. The ejaculate was transferred to laboratory immediately and diluted with pre-warmed semen extender (Optixcell, France) (1:1). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and divided into 5 groups according to BHT concentration (0, 0.5, 1.0, 2.0 and 4.0 mM) and cryopreserved in LN2 tank until evaluation. Frozen-thawed semen was transferred to 1.5 ml tube and incubated for 0, 2 and 4h. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain). There were not significant effects of BHT supplementation (0, 0.5, 1.0 and 2.0 mM) on total motile and VSL with 25μm≥ at 0, 2 and 4h. However, 4.0 mM of BHT supplementation showed negative effect on total motile (26.3%), VSL with 25μm≥ (1.3%) at 0 h (p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method and divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). There were no significant differences of LIA, LDA, DIA and DDA on various BHT concentrations at 0 and 2 h. However, 4.0 mM BHT supplementation showed decreased LIA compared with 0, 0.5, 1.0 and 2.0 mM BHT at 4 h (34.6, 37.1, 43.6, 45.4 and 14.7% vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.01). Addition of 4.0 mM of BHT showed negative effect on plasma membrane integrity compared with that of 0, 0.5, 1.0 and 2.0 BHT at 2 h (71.9, 64.2, 64.6, 67.5 and 31.7 % vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.05). In conclusion, various BHT concentrations on optixcell extender showed no improvement on sperm motility, viability and plasma membrane integrity.

Severe combined immune deficiency (SCID) pig is very important research model for biomedical research, such as the development of humanized tissues and organs for transplantation and long-term evaluation of transplanted cancer or stem cell of human origin. FOXN1 gene encodes a transcription factor essential for the development and function of thymic epithelial cells (TECs), the primary lymphoid organ that supports T-cell development and selection. In this study, we are going to produce the FOXN1 KO SCID pigs using the Crispr/Cpf1 method. Porcine genomic DNA sequences were analyzed and the target sequences were selected using a web tool, Benchling (https://benchling.com/). The designed crDNA oligos was synthesized by the Oligonucleotide Synthesis Service (Macrogen Inc., Seoul, Korea). To generate the AsCpf1-mCherry-Puro construct, pTE4396 (#74041; Addgene, Cambridge, MA, USA) was modified by removing the NeoR/KanR sequence using BstBI and SmaI. Then, the mCherry-Puro sequence from pSicoR-Ef1a-mCh-Puro (#31845; Addgene, Cambridge, MA, USA) digested with the same restriction enzymes was inserted into the aforementioned NeoR/KanR-deleted vector. The crDNA #1 or crDNA #2 was inserted into the pTE4396 and AsCpf1-mCherry-Puro vectors in the U6 promoter region using BsmBI enzyme, respectively. The two vectors were transfected with lipofectamine 3000 (Life Technologies, Grand Island, NY, USA) and selected with puromycin and G-418 antibiotics. As a result, we established a cell line into which two vectors (pTE4396+crFOXN1#2 and AsCpf1- mCherry-Puro+ crFOXN1#1) and were inserted. Further studies are needed to characterize FOXN1 KO cell lines.

Lysophosphatidic acid (LPA) is an important signaling molecule which mediates many different cellular responses. The purpose of this study was to investigate the effect of in vitro culture (IVC) medium supplemented with LPA on the preimplantation embryonic development of porcine embryos derived from in vitro fertilization (IVF). Embryos derived from IVF were cultured in PZM-3 medium supplemented with 30 μM LPA on Day 1 to Day 7, Day 1 to Day 3 (early stage), or Day 4 to Day 7 (late stage), or without LPA. Moreover, the messenger RNA (mRNA) expression of obtaining blastocysts from each group were analyzed. Data were analyzed by ANOVA followed by Duncan using SPSS (Statistical Package for Social Science) mean ± SEM. There was a significantly higher cleavage rate in Day 1 to Day 7 than control (71.25% and 57.46%, respectively) and significantly higher total cell number of blastocysts in Day 1 to Day 3 and Day 4 to Day 7 than control (56.07,56.53 and 45.19, respectively). The results also showed that the mRNA expression level of PCNA, Bcl-2 and Bax in Day 1 to Day 7 group blastocysts were significantly higher than control and the expression level of Bax in Day 1 to Day 3 was also significantly higher than control. Moreover, it also showed that Bcl-2/Bax mRNA ratio in D1-3 group was significantly lower than control but D4-7 and D1-7 groups were comparable to control group. In conclusion, our results suggest that treatment with 30 μM LPA during IVC improves the porcine early embryo cleavage and the blastocyst total cell number after IVF and regulating the mRNA expression of blastocysts during blastocyst formation.

Recently, we published a microinjection method for generating transgenic cattle using the DNA transposon system and their analysis by next-generation sequencing (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In that study, we generated transgenic cattle using two different types of DNA transposon system, sleeping beauty (SB) and piggybac (PB), carrying Yellow fluorescent protein with SB (SB-YFP, female) and green fluorescent protein with PB (PB-GFP, male) under the control of the ubiquitous CAG promoter, respectively. The female and male founder cattle have been grown up to date (the female age: 40 months old, the male age: 33 months old) without any health issues. In genomic instability and blood analysis, there was no significant differences between wild type and founder cattle. In the present study, we confirmed germ-line transmission of the transposon-mediated transgene integrations and ubiquitous and persistent expression of transgene in second generation of offspring (F1). The F1 was born without any assistance and expressed GFP in the eyes without UV light. The ubiquitous expression of GFP was detected in skin fibroblast from the ear tissue and confirmed by genomic DNA PCR, which suggest that the transgene from the PB-GFP was successfully transmitted. Unfortunately, no transgene from SB-YFP were identified. To confirm the transgene integration site, the genomic DNA from blood was extracted and performed next-generation sequencing (NGS). The GFP gene was integrated in chromosome 4 (two copies), and 6. As results, a total of two copies of paternal transgene transmitted into the F1. All the integrated position was not related with coding region and there was no significant difference in genomic variants between transgenic and non-transgenic cattle. To our knowledge, this is the first report of germ-line transmission through non-viral transgenic founder cattle. Those transgenic cattle will be valuable resource to many fields of biomedical research and agricultural science.

Several studies on the correlation between temperament and genetic diversity are conducted in animals as well as human. Horse temperament is especially important because it is important factor for horse riding and racing. In this study, we performed targeted exome sequencing to find single nucleotide polymorphisms (SNPs) served as genetic markers that can evaluate the aggressive and docile levels of horses. We selected 71 candidate genes related to animal and human temperament through previous researches and verified it on the human reference genome (hg38) and horse reference genome (equCab2). We found that 16 orthologous genes were present in horse reference genome and 17 homologous genes found in horses based on the human reference genome. Finally, we designed probes to find the genetic variation in selected 33 genes. The sequencing libraries were constructed using the designed probe and DNA samples extracted from the blood of 8 aggressive and 8 docile horses. The constructed libraries were sequenced using the Illumina Hiseq2500 platform. SNPs data obtained from targeted exome sequencing will be used for genome wide association study (GWAS) and Sanger sequencing validation. This study will help to assess the horse temperament and to select superior horses for riding or racing.

Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.

Previously, we reported a quantitative trait locus (QTL) that affect total teat number (TTN) on pig chromosome 7 (SSC7) in a large F2 intercross population between Landrace and Korean native pigs. The aim of this study was to refine the QTL associated with TTN and to identify positional candidate gene(s) within the refined genomic region. TTN was recorded in 1,105 F2 progeny. All experimental animals were genotyped using 998 informative single nucleotide polymorphism (SNP) markers located on SSC7. A haplotype-based linkage and association analysis using the PHASEBOOK programme was applied to perform high-resolution QTL analysis. Additionally, linear mixed-effect models were used to assess the effect of a positional candidate gene on TTN and other economically important traits [i.e., thoracic vertebrae number (THO), carcass body length (CBL) and weight (CW), back fat thickness (BFT) and intramuscular fat content (IMF) in loin muscles]. Joint linkage and association analysis refined the critical region to a 1.07 Mb region that included a novel positional candidate gene, BRMS1L, that encodes the breast cancer metastasis-suppressor 1-like protein, which could possibly be implicated in normal mammary gland development. Significant association of an SNP marker (g.-1087 G>A) in the 5’-flanking region of BRMS1L with TTN (P=1.10x10-8), THO (P=5.80x10-4), and CBL (P=0.038) was observed. Based on these data, we propose BRMS1L as a positional candidate gene for TTN in pigs. After validation of the association in other independent populations and further functional studies, these results could be useful in optimizing breeding programmes that improve TTN and other economically important traits in swine

Alterations affecting the status of robustness and health can bring about physiological changes including hematological parameters in pigs. To identify quantitative trait loci (QTL) associated with 8 hematological phenotypes (one leukocyte trait, six erythrocyte traits, and one platelet trait), we performed a genome-wide association study using the Porcine SNP 60K BeadChip in an intercross population between Landrace and Korean native pigs. A total of 36,740 SNPs from 816 F2 offspring were analysed for each blood related traits after filtering by quality control. Data were analysed genome-wide rapid association using the mixed model and regression (GRAMMAR) approach. A total of 257 significant SNPs (P<1.36x10-6) on SSC3, 6, 8, 13, and 17 were detected for blood related traits in this study. Interestingly, the genomic region between 17.9 and 130 Mb on SSC8 was found to be significantly associated with RBC, MCV, and MCH. Our results include 5 significant SNPs within five candidate genes (KIT, IL15, TXK, ARAP2, and ERG) for hematopoiesis. Further validation of these identified SNPs could give valuable information for understanding the variation of hematological traits in swine.

Porcine litter size is a quantitative trait and its heritability is especially low. So it is necessary to identify porcine reproductive gene and protein. The establishment of pregnancy requires performance of a receptive endometrium. We analyzed the endometrial tissue protein of porcine and would find out biomarker proteins related to porcine litter size. We sorted the two groups according to litter size of porcine: a small litter size group (SLSG) (n=2) and a large litter size group (LLSG) (n=2). The porcine endometrial tissue samples were analyzed separately using 2-dimensional electrophoresis (2-DE) within the isoelectric point ranges of 3.0 to 10.0, and then differential proteins were identified using MALDI-TOF analysis. In comparison of SLSG(small litter size group) with LLSG(large litter size group), a total of 9 protein spots differentially expressed on porcine endometrial tissue 2-DE gels, among which 5 spots were up-regulated proteins as retinol dehydrogenase 16-like isoform 1, Acrosin-binding protein, alpha-N-acetylgalactosaminidase. phosphoglycerate kinase 2, Acrosin-binding protein in LLSG. And 4 spots were up-regulated proteins as phosphoglycerate kinase 2, prenylcysteine oxidase in SLSG.

An intensive analysis was conducted of the maternal lineages of the Jeju native pigs (JNPs). A total of 100 mtDNA sequences from Asian wild boars (AWB), European wild boars (EWB), Asian domestic pigs (ADP), European domestic pigs (EDP), and JNPs were used for the phylogeny and network analyses. Two distinct JNP groups were found JNPA and JNPE in the Asian and European cluster. The maternal lineage of JNPE was the closest to that of EWB and a clear haplogroup sharing an identical haplotype (hap16) among 15 individuals of JNPE and 2 individuals of EWB was identified. However, except for hap18, no EDP shared any identical haplotypes with JNPE, suggesting that no obvious maternal contribution of EDP has occurred in JNPE in recent years. The possible existence of an additional and unknown path of maternal lineage from EDP into JNPE could therefore be postulated, in addition to those from AWB and ADP into the JNPA groups. Thus, JNPE appeared to have a pure maternal lineage that had no recent contact with EDP, and both the JNPA groups and JNPE are pure Jeju native pigs.

Embryo development is very important in reproductive physiology of domestic animal experiments. Therefore, in the above experiment, we want to provide a lot of important information with regard to fertilization breeding by looking at the expression of transcription factor by early embryo development. It is known that mice affect early embryonic development of many transcription factors, many experiments are underway. Different types of mammals showed different expression patterns, thus, we used pigs, which are known to be the most similar to humans, to observe the expression of transcription factors in early embryonic development. Transcription factors were observed using CDX2, OCT4 and E-CADHERIN. CDX2 was expressed in 2 cells, OCT4 and E-CADHERIN were expressed in blastocyst. OCT4 was expressed specifically in ICM (inner cell mass) in blastocyst, and E-CADHERIN was expressed in cell wall and junction of blastocyst. These results show that CDX2, OCT4 and E-CADHERIN play an important role in early embryonic development in pigs.

한우농가의 사육기반을 확보하기 위해서는 번식관리를 통해 송아지 생산율을 향상시킬 필요가 있다. 본 연구는 소규모에서부터 대규모 농가 사육 조건에서의 번식우 관리의 현황을 파악하므로 서 한우 송아지 생산율을 향상 방안을 찾기 위하여 2016년 하반기에 농가 실태조사를 실시하였다. 조사 대상은 총 45농가였으며, 지역별로 남부(경남), 중부(경북, 충남), 북부(경기, 강원)로 나누었 고, 사육 마릿수에 따라 소(20마리 이하), 중(20〜50마리), 대(50마리 이상)규모로 나누어 진행했다. 총 조사 두수는 번식우로 사용하는 가임암소 1800여두를 대상으로 하였다. 조사에 의하면 조사대 상 농가의 초임월령은 평균 20.9개월로 조사되었다. 최초 분만월령은 평균 28.7개월령 이었으며, 수태당 인공수정 횟수는 1.45회였다. 수태율은 소규모 농가가 75.2%로 중규모(70.6%)나 대규모 (71.4%)보다 높은 것으로 나타났으며 평균 72.4%였다. 분만 후 인공수정일수는 평균 119.8일로 조 사되었다. 수태율과 BCS와의 관계에서 소규모 농가의 수태율이 가장 높았으며(75.2±16.9%), BCS는 소규모 농가가 3.1로 중규모(3.0) 또는 대규모 농가(2.8)보다 높았다. 일반농가에서는 송아지 생산율 이 67.64%였으나 발정관찰 보조기구를 사용하는 농가는 78.06%로 송아지 생산율이 10.42% 높았 다. 농가여건에 따라 별도의 운동장 또는 방목을 사용하는 경우 수정율이 92.54%였고, 사사관리만 하였을 때 89.07%였고, 분만율도 운동장 사용시 88.12%로서 사사관리의 69.83%보다 약 18.3% 향 상되는 것으로 나타났다. 번식우를 대상으로 예방접종(IBR, BVDV)을 실시한 농가는 분만율이 79.67%로서 그렇지 않은 농가의 69.27%에 비해 10.4%가 높았고, 또한 분만시 유사산·폐사율도 예 방접종 농가는 5.36%인데 비해 미실시 농가에서는 9.77%로 예방접종을 하였을 때 4.41% 감소한 것으로 나타났다. 따라서 번식우는 충분한 운동과 발정관찰에 노력해야 하고 예방접종을 하므로서 분만율 향상 등 송아지 생산과 육성율에 효과적인 것으로 나타났다.