Secretory leukocyte protease inhibitor (SLPI), also known as neutrophil elastase and cathepsin-G protease inhibitor, functions in protection of epithelial cells from proteases. SLPI is expressed and secreted by many mucosal tissues, including lungs, seminal vesicles and cervix in women. SLPI plays an important role in protection of endometrial epithelial cells during pregnancy from degradation by degradation by proteases derived from trophoblast at the maternal-conceptus interface. In pigs, SLPI mRNA is known to be expressed in endometrial tissues, but the expression of SLPI in the endometrium throughout the estrous cycle and pregnancy has not been determined. Therefore, we analyzed the expression and regulation of SLPI mRNA in the endometrium throughout the whole stages of the estrous cycle and pregnancy in pigs. We obtained endometrial tissues from gilts on Days 0 (day of estrus), 3, 6, 9, 12, 15, and 18 of the estrous cycle and on Days 10, 12, 15, 30, 60, 90, and 114 of pregnancy. Real-time RT-PCR analysis showed that the expression of SLPI mRNA in the endometrium increases during midt-o late pregnancy. During the estrous cycle, levels of SLPPI mRNA in estrus and proestrus were higher than those in diestrus and metestrus. In situ hybridization analysis showed that SLPI mRNA was specifically localized to the glandular epithelial cells in the endometrium during pregnancy with strong signal intensity during mid-to late pregnancy. SLPI mRNA was not detectable in conceptus tissues on Days 12 and 15 of pregnancy, but SLPI mRNA was expressed in chorioallantoic tissues during mid-to term pregnancy with increasing levels toward term pregnancy. To determine the effects of steroid hormones, estrogen and progesterone, on the expression of SLPI mRNA, endometrial explant tissues from immature pigs were treated with increasing doses of estradiol-17β (E2) and progesterone (P4). Increasing doses of E2 and P4 increased the expression of SLPI mRNA in endometrial tissues. These results showed that SLPI was expressed in the endometrium in a pregnancy stage-and cell type-specific manner and the expression of SLPI was regulated by E2 and P4 in endometrial tissues, suggesting that SLPI may play an important role in regulating the endometrial epithelial cell function during mid-to late pregnancy in pigs. Further analysis to determine the roles of SLPI at the maternal-conceptus interface is still needed.

Caspases are a family of cysteine protease enzymes composed of more than 10 members that play essential roles in apoptosis and inflammation. It has been reported that caspases play a critical role in regulating apoptosis at the maternal-conceptus interface in many species. However, the expression and regulation of caspases have not been determined in the endometrium in pigs. Therefore, we analyzed the expression, localization, and regulation of caspases in the endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy. The expression of CASP6, CASP7, and CASP8 during the estrous cycle and CASP3, CASP6, CASP7, and CASP10 changed during pregnancy. Levels of CASP3 mRNA in the endometrium were higher on Day 12 of pregnancy than the estrous cycle and levels of CASP7 mRNAs were highest on Day 15 of estrous cycle and pregnancy. Immunohistochemistry analysis showed that CASP3 protein was localized to endometrial epithelial cells on Days 12 and 15 the estrous cycle and pregnancy, but cleaved CASP3 was localized only to luminal epithelial (LE) cells on Days 12 and 15 of pregnancy in the endometrium. CASP7 protein was localized to endometrial LE cells only on Day 15 of pregnancy. CASP3, CASP6, CASP7, CASP8, and CASP10 mRNAs were detectable in conceptus on D12 and D15 of pregnancy, and chorioallantoic tissues expressed CASP3, CASP6, CASP7, CASP8, CASP8, and CASP10 with increasing levels toward term pregnancy, except CASP3 mRNA. The effect of steroid hormones and interleukin-1βß (IL1B) on CASP3 expression and the effect of interferon-γ(IFNG) on CASP7 expression was determined by endometrial explant cultures and we found that CASP3 expression was increased by IL1B and CASP7 expression was increased by IFNG in a dose-dependent manner. These results showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy in a stage- and/or pregnancy-specific dependent manner and some caspases were regulated by IL1B or IFNG in the endometrial tissues, suggesting that caspase may play an important role in regulating apoptosis for the establishment and maintenance of pregnancy at the maternal-conceptus interface in pigs.

Although some factors, including season, age, type of estrus (natural estrus vs. induced estrus) and semen type (conventional vs. sexed), affect the conception rate following artificial insemination (AI) in dairy cattle, there is little information about the influence of ovarian characteristics, such as preovulatory follicle (PF) location at estrus, on fertility in dairy cattle. In most breeds of cattle the right ovary appears to function more actively than the left and about sixty percent of pregnancies in dairy cattle occur in the right horn of uterus (Reece and Tuner, 1938). Our study aimed to compare conception rates in dairy cattle between PFs that developed in the left ovary and those that developed in the right ovary at estrus. In this study, we examined the locational effect (left or right ovary) of the preovulatory follicle (PF) on fertility in dairy cattle. In total, 955 artificial inseminations (AI) were analyzed. At AI, PF locations were examined using rectal palpation, and dairy cattle were divided into two groups on their PF locations: (i) the PF located in the left ovary (L-PF); and (ii) the PF located in the right ovary (R-PF). Pregnancy was diagnosed by rectal palpation or ultrasonographic examination 60 days after AI. The conception rate was 38.1% in all dairy cattle. Conception rate was higher in the R-PF (40.8%) than in the L-PF (33.2%). In summary, PF development in the right ovary was associated with increased conception rates in dairy cattle.

The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

The objective of this study was to evaluate the effect of treating dairy cattle with exogenous human chorionic gonadotrophin (hCG), five (5) days post artificial insemination (AI) on serum progesterone (P4) concentration and pregnancy rate. In this experiment, five days after AI, cows were assigned randomly to two groups namely: a) treated group (67) which were administrered with 1500 IU hCG (Chorulon) and b) control group (61), which received no treatment. On day 5, 10, 15 and 20 after the artificial insemination, blood samples from a total of 8 cows (4 from each group) were collected and were analyzed for serum P4 concentration. Cows were detected for estrus according to standing heat by visual observation. Cows that were detected still in estrus after days 18-24 were re-inseminated and recorded as not pregnant (open). Pregnancy diagnosis was conducted by ultrasonographic examination and transrectal palpation of the uterus on approximately 60 days in cows that observed to be not in estrus. The conception rate in hCG treated and control groups were 52.5 and 36.1%, respectively. The results proved that there were no significant differences in conception rate between two groups (p=0.0568). However, pregnancy rates were reduced by hCG treatment. Average serum P4 concentrations did not differ between Hcg-treated and control groups on day 5 (0.377 versus 0.375 ng/ml). On day 20 serum P4 concentrations were greater in the treated group compared with the control group (3.085 versus 2.010 ng/ml). The treatment with hCG seemed to increase P4 level compared with the control. In conclusion, the results of this study showed that 1500 IU of hCG administered on 5 day post AI increased conception rate in dairy cows. This was supported by the results on serum P4 concentration which was greater in hCG treated group.

The major focus of this study is to analyze the expression of bovine MMPs and to monitor their activity during the estrus cycle and pregnancy. During pregnancy, MMP-2 expression was detectable around 30 days but became insignificant by 60 days, then started to increase again around 90 days and reached the maximum at 250 days. The activity of MMP-2 protein changed in accordance with its expression level. As expected, the level of TIMP-2 exhibited a reverse pattern. About MMP-9, high level expression was observed as early as 30 days and gradually increase until 90 days. Then started to decrease after 250 days. Again, the sites of MMP-9 expression were similar to those of MMP-2. On the other hand, expression of TIMP-3 remained low until 90 days but showed a small and temporal increase around 250 days. In summary, expression of different MMPs were differentially regulated during estrus cycle and pregnancy. While the expression of MMP-2 was high in estrus cycle, MMP-9 slowly takes over with the progression of pregnancy. These results indicated that the luteal tissue perform distinct functions during pregnancy and estrus. Perhaps the activity of MMP-2 is required for the structural remodeling of luteum, resulting the suppression of P4 inflow from blood. On the other hand, steady maintenance of MMP-9 throughout luteal development is important for the activation of cell proliferation, maturation and angiogenesis.

Assays for detecting pregnancy-associated glycoproteins (PAGs) levels in the blood of pregnant cattle have been developed and commercialized to determine pregnancy status. This study was conducted to determine the accuracy of a PAG-based ELISA test for pregnancy detection in Hanwoo. The anticoagulated treated whole blood was sampled from 595 Hanwoo that were 28-71 d (mean 49) and 85-128 d (mean 106) after artificial insemination. At the 2nd blood collection time, a transrectal palpation test was used as the standard for pregnancy diagnosis. For PAGs and the transrectal palpation test (n=595), the sensitivity was 98.4% (pregnancy, 481/489), the specificity was 100% (open, 106/106), and the accuracy was 98.7% (pregnancy+open, 587/595). In conclusion, the rapid PAG-based ELISA pregnancy test was highly sensitive and specific for pregnancy detection in Hanwoo.

The artificial insemination (AI) is one of the best assisted reproductive technologies for increasing reproductive capacity and facilitating the genetic improvement in farm animals. AI has been used in Uganda for over 60 years, but a small population of the total herd has been used. This study was conducted to investigate the efficacy of AI with estrus synchronization technique and to propose ways of improving the productivity of dairy farms through AI services in Uganda. In total, 78 cows from 11 dairy farms were selected for timed-AI. Synchronization was performed according to the ovsynch programs followed by AI using frozen semen from Korean Holstein (0.5 ml straws). Pregnancy rate was varying among farms (0-50%) and the overall pregnancy rate was 28.2%. Cows in luteal phase at the time of treatment was 40.0% whereas that in follicular phase was 20.8%. After treatment, cows that showed normal estrus signal were 45.5% (25/55). Abnormal estrus was categorized into pre-estrus (9.1%), cystic ovaries (21.8%), anestrus (18.2%) and delayed ovulation (5.5%), respectively. These results imply that an assured protocol for timed-AI should be developed to improve the productivity of dairy farms through AI services in Uganda.

Embryo transfer (ET) could be a relevant tool for genetic improvement programs in horses similar to those already underway in other species and produce multiple foals from the same mare in one breeding season. However, there have been no reports describing equine embryo transfer performed in Korea. In the present study, we performed an equine embryo collection and transfer procedure for the first time. We examined the embryo collection and pregnancy, size of embryo during the incubation period after collection, and progesterone (P4) and estradiol-17ß (E2) concentrations in mare’s serum at embryo collection and transfer. A total of 16 donors responded to estrus synchronization; estrus was induced in 12 donors and 4 recipients, and artificial insemination was successful in 10 donors and six blastocysts were collected from donors. Of these blastocysts, we monitored the size of blastocysts for 3 day during incubation and transferred 2 blastocysts to a recipient, with 1 successful pregnancy and foal achieved. The dimensions of equine embryo at day 7 to day 9 were 409 μm, 814 μm and 1,200 μm. The serum P4 and E2 concentrations were 7.91±0.37 ng/μL and 45.45±12.65 ng/μL in the donor mare, and 16.06±3.27 ng/μL and 49.13±10.09 ng/μL in the recipient mare.

The successful establishment and maintenance of pregnancy is achieved by well-coordinated interactions between the maternal uterus and the implanting conceptus. In pigs, the conceptus undergoes dramatic morphological and functional changes, and secretes various biological products such as estrogens and cytokines, interleukin-1beta (IL1B), interferon-gamma (IFNG), and IFN-delta (IFND) during the implantation period. The uterine endometrium in response to the conceptus-derived molecules and ovarian progesterone becomes receptive to the conceptus by changing cell adhesion molecule expression, epithelial cell depolarization and secretory activity. Conceptus-derived estrogen acts as the maternal pregnancy recognition signal which changes the direction of endometrial prostaglandin (PG) F2 secretion from the uterine vasculature into the uterine lumen. Estrogen also induces the expression of a variety of endometrial genes, including AKR1B1, FGF7, LPAR3, and SPP1. The function of cytokines, IL1B, IFNG, and IFND, in the endometrium is not fully understood, but some recent work shows that IL1B is involved in the synthesis and transport of endometrial PGs by regulating endometrial expression of PG-synthetic enzymes, PTGS1, PTGS2, and AKR1B1, and PG transporters, ABCC4 and SLCO2A1. Estrogen and IL1B also stimulate endometrial expression of IFN signaling molecules, suggesting that estrogen and IL1B act cooperatively on priming the endometrial function of conceptus IFNG and IFND. In turn, IFNG derived from the elongating conceptuses, induces many endometrial genes, including CXCL9, CXCL10, CXCL12, and SLA-DQ. The role of IFND at the maternal-conceptus interface is not well understood yet. Further analysis of the molecules derived from the endometrium and conceptus will provide insights into the cellular and molecular basis of maternal-conceptus interactions for the establishment of successful pregnancy in pigs.

담관낭종은 담도가 비가역적으로 확장된 선천성 질환으로 담도암의 발생 위험으로 인해 수술적 절제를 시행하는 것이 원칙이다. 임신 중에는 호르몬의 영향 및 확대된 자궁, 증가된 복압 등으로 담관낭종이 증상을 일으킬 수 있다. 그러나 임신 중에는 생리적 변화로 나타나는 임신 자체의 증상과 낭종에 의한 증상을 구분하기가 어렵고, 임신 중 시행할 수 있는 영상 검사가 제한되어 있어 담관낭종의 진단이 지연됨으로써 산모와 태아에게 치명적인 결과를 초래할 수 있다. 또한 수술로 인한 태아와 산모의 이환율과 사망률을 고려할 때, 즉각적인 외과 수술보다는 담관낭종으로 인한 합병증을 예방하고 출산 시까지 산모와 태아를 안전하게 보호하기 위한 단계적인 치료 전략이 중요하다. 저자들은 임신 중 담관낭종에 의해 발생한 황달 및 복통으로 내원한 26세 산모에서 투시 조영술 도움 없이 내시경 초음파를 이용한 담도 배액술을 시행하여 증상 완화와 함께 최대한 임신을 유지할 수 있었다. 이후 성공적인 출산 및 산후 회복기를 거쳐 담관낭종을 수술적으로 제거한 1예 를 경험하였기에 문헌고찰과 함께 보고하고자 한다.

This study was carried out to evaluate the influencing factors that affect the reproductive performance of cows at the Monirampur upazila in Jessore district of Bangladesh. A total of 224 cows were brought to the upazila livestock hospital for artificial insemination (AI). The cows were inseminated between 12 to 18 hours from the onset of estrus and data was obtained from the owner. Out of 224 cows, 133 became pregnant and 91 were non pregnant. In this study, the overall pregnancy rate was 59.29%. Among the age variability, the highest pregnancy rate (70.27%) was at the age of 4 years old. In case of breed variation, the highest pregnancy rate was observed in local breed (69.07%) compared with other crossbred cows. Hence the breed variations significantly influence the conception rate of cows. According to the parity, we found that the pregnancy rate was increasing with their parity but decreasing after 4th parity. The highest conception rate was observed in 3rd parity (67.74%) which was significantly higher than that of heifers (Parity-0). Here we also found that the types of bull semen used for AI had no significant effect for pregnancy rate. The skills of AI technician for AI to cows were significantly affecting the pregnancy rate. However, this study is not enough for rating and comment about the reproduction performance of cows. Therefore, further extensive study is needed for rating and recommendation for the cattle up gradation at that particular area.

The migration, adhesion, and proliferation of conceptuses during pregnancy are tightly controlled processes that are mediated by various factors including cytokines, growth factors, and hormones. Among many factors, chemokines play key roles in lymphocyte trafficking, cellular proliferation, vascularization, and embryogenesis in many mammalian species. Especially, it has been shown that C-X-C chemokine ligand 12 (CXCL12) plays an important role in early pregnancy by promoting trophoblast invasion, proliferation, and differentiation through its receptor, C-X-C chemokine receptor 4 (CXCR4) in humans. However, expression and function of CXCL12 in the uterine endometrium during pregnancy have not been well studied in pigs. Thus, we determined expression of CXCL12 and its receptor, CXCR4, in the uterine endometrium during the estrous cycle and pregnancy in pigs. We obtained endometrial tissues from gilts on day (D) 12 and D15 of the estrous cycle and D12, D15, D30, D60, D90, and D114 of pregnancy, conceptus tissues from D12 and D15 of pregnancy, and chorioallantoic tissues from D30, D60, D90, and D114 of pregnancy. Real-time RT-PCR analysis showed that levels of CXCL12 and CXCR4 mRNAs changed in the uterine endometrium during pregnancy. Levels of CXCL12 and CXCR4 mRNAs on D15 of pregnancy were higher than those on D15 of the estrous cycle. After D15 of pregnancy levels of CXCL12 and CXCR4 mRNAs gradually decreased toward term of pregnancy, and CXCL12 and CXCR4 were expressed in the chorioallantoic tissues during the mid- to late pregnancy. CXCL12 and CXCR4 mRNAs were expressed in chorioallantoic tissues during mid- to late pregnancy, and RT-PCR analysis showed that CXCL12 and CXCR4 mRNAs were detectable in conceptus on D12 and D15 of pregnancy. Immunohistochemistry showed that CXCL12 proteins were localized to endometrial luminal and glandular epithelial cells during the estrous cycle and pregnancy, and to chorionic epithelial cells during mid- to late pregnancy. Abundance of CXCL12 mRNAs, but not CXCR4, in the uterine endometrium was increased by the treatment of IFNG. These results showed that CXCL12 and CXCR4 were expressed in the uterine endometrium, conceptus, and chorioallantoic tissues and IFNG increased endometrial CXCL12 expression in pigs, suggesting that CXCL12 and its receptor may play a key role in regulation of the establishment and maintenance of pregnancy by affecting the conceptus development in pigs. [supported by the Next Generation BioGreen 21 Program (#PJ01110301), Rural Development Administration]

For the establishment and maintenance of successful pregnancy the maternal immune system must tolerate semi-allogenic fetus during pregnancy. Several mechanisms explaining immune tolerance have been proposed. Among those, it has been suggested that the CD40/CD40L system is involved in immune tolerance in several tissues. However, expression and function of CD40/CD40L in the maternal-fetal interface during pregnancy have not been studied in pigs. Thus, this study determined expression and localization of CD40 and CD40L in the uterine endometrium during pregnancy in pigs. We obtained uterine endometrial tissue samples from day (D) 12 and D15 of the estrous cycle and from D12, D15, D30, D60, D90 and D114 of pregnancy. Quantitative real-time PCR analysis showed that levels of CD40L mRNA expression during pregnancy increased on D15 of pregnancy and decreased thereafter whereas levels of CD40 mRNA was highest on D30 of pregnancy. Localization of CD40 and CD40L proteins by immunohistochemistry showed that CD40 was localized to vascular endothelial cells with strongest signal intensity on D15 of pregnancy, and CD40L was localized to luminal epithelial cells on D15 of pregnancy and amniotic membrane during mid- to late pregnancy. To determine the effect of IFNG on CD40 and CD40L expression, we took advantage of endometrial explant culture using tissues from D12 of the estrous cycle, and found that CD40 was up-regulated by IFNG in a dose-dependent manner. These results showed that CD40 and CD40L were expressed in the uterine endometrium in a cell-type and stage-specific fashion during pregnancy, and IFNG induced CD40, indicating that the CD40/CD40L system may be important for establishment and maintenance of pregnancy in pigs. [Supported by the Next Generation BioGreen21 Program (#PJ01110301), Rural Development Administration]

High reproductive efficiency is a prerequisite for dairy animals in order to optimize dairy production. An accurate and early pregnancy diagnosis is a crucial aspect for better reproductive management in livestock. The indigenous, visual and clinical methods have various limitations including accuracy, sensitivity, specificity, later stages of applicability and requires highly skilled manpower. This hurdles the researchers to ensue further research on developing novel early pregnancy diagnostics for dairy animals. However, the advantage of molecular techniques like proteomics has given a new hope to look for pregnancy biomarkers in animal research. In this study, we assayed the pregnancy-associated glycoprotein (PAG) levels using anti-bPAG antibody. Serum plasma samples were collected 30, 60, 90 and 120 days after insemination. Cattle diagnosed pregnant with singleton are used for this experiment in which blood samples were collected. The plasma PAG level is gradually increased after insemination to until 60 days and drastically decreased after that. The PAG level was approximately 2-fold greater during 30 days compared with 60 days. These results concluded to validate our other pregnancy diagnosis methods including circulating microRNA and cell free DNA concentration (data not shown). Despite this proteomic approach, validating all other molecular technique results may give the exact time for early pregnancy diagnosis.

Early pregnancy results in th production of various signal molecules such as steroids, prostaglandins, and many protein factors. The proteins especially produced by the placenta have been used to detect pregnancy for many years in other species. More recently, pregnancy-specific protein B, which is a placental glycoprotein can be measured by RIA or proteomic methods in serum of pregnant cow. And 2D Fluorescence difference gel electrophoresis (DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. For this reason, we are analyzed serum of bovine. The purpose of this study was to apply DIGE technique for identification of bovine pregnancy-specific proteins using bovine pregnant and non-pregnant serum samples. Serums of 2 pregnant Holstein dairy cattle at day 21 after AI and those of 2 non-pregnant were used in this study. The molecular weight and charge matched cyanine dyes enable pre-electrophoretic labeling of non-pregnancy and pregnant serum proteins which are then mixed and labeled with Cy2 were used as an internal standard. Two pools of proteins are labeled with Cy3 and Cy5 fluorescent dyes, respectively. Labeled proteins with Cy2, Cy3 and Cy5 mixed together and separated in same gel and then were detected by fluorescence image analyzer. The 2D DIGE analysis using fluorescence CyDye flour showed higher sensitivity and better reproducible results than conventional 2D gel electrophoresis. Approximately 1,500 protein spots were detected by 2D DIGE. The differentially expressed proteins were identified by MALDI-TOF Mass spectrometer. Total 16 protein spots differentially expressed in the pregnant serum were detected, among which 7 spots were up-regulated proteins identified as conglutinin precursor, modified bovine fibrinogen, IgG1 etc, and 6 spots were down-regulated proteins identified as hemoglobin, complement component 3, bovine fibrinogen, IgG2a etc. These results indicated that DIGE system could be advantageous for the analysis of serum proteomics diversified by physiological conditions.

Embryo transfer (ET) has been applied to many species, which also useful tools for genetic improvement in horses. ET enables to produce a multiple foals from one donor mare in the same breeding season. However, there have been no reports describing equine embryo transfer performed in Korea. In the present study, we performed an equine embryo collection and transfer procedure for the first time. We examined the embryo collection and pregnancy, size of embryo during the incubation period after collection, and progesterone (P4) and estradiol-17ß (E2) concentrations in mare’s serum at embryo collection and transfer. A total of 16 donors responded to estrus synchronization; estrus was induced in 12 donors and 4 recipients, and artificial insemination was successful in 10 donors and six blastocysts were collected from donors. Of these blastocysts, we monitored the size of blastocysts for 3 day during incubation and transferred 2 blastocysts to a recipient, with 1 successful pregnancy and foal achieved. The dimensions of equine embryo at day 7 to day 9 were 409 μm, 814 μm and 1,200 μm. The serum P4 and E2 concentrations were 7.91±0.37 ng/μL and 45.45±12.65 ng/μL in the donor mare, and 16.06±3.27 ng/μL and 49.13±10.09 ng/μL in the recipient mare.

The objective of this study was to determine the effect of different pregnancy stages on the eating and ruminating behavior of Hanwoo heifers (Bos taurus coreanae). A total of twelve Hanwoo heifers were divided into four groups depending on pregnancy stage: C: before pregnancy, 9 months old, T1: 3 months pregnant, 16 months old, T2: 6 months pregnant, 19 months old, T3: 9 months pregnant, 22 months old (3 replicates per group). Concentrate of 1% of the body weight was fed to them. Roughage was provided at ad libitum to all treatments. Our results revealed that the intake of roughage was the highest in the T3 group (at 3.68 kg) and the lowest in the C group (at 1.02 kg, p<0.05). Total intake in the order from high to low was: T3 > T2 > T1 > C (p<0.05). The rate of dry matter intake compared to body weight (DMI/BW, %) from high to low order was: T3 > T2 > T1 > C (p<0.05). Eating time, ruminating time, and chewing time were the highest in T3 group (p<0.05). Resting time was the highest in C group (p<0.05). The number of bolus and the number of total chews were significantly higher in T3 compared to those in other groups. However, ruminating time per bolus was not significantly difference among groups. The number of chews per bolus from high to low order was: C > T1 > T3 > T2 (p<0.05). The number of bolus per minute was not significantly different among groups. The number of drinking and the number of defecating were significantly higher in C and T1, respectively (p<0.05). However, the number of urinating had no effect according to pregnancy stage. Eating rate and chewing efficiency were significantly higher (p<0.05) in accordance with pregnancy stage (T3 > T2 > T1 > C). Ruminating efficiency was significantly (p<0.05) higher T1 than that in other groups.

Raising replacement heifers is important aspect of dairy farm management that can be characterized as a long-term performance and high-cost production period. This study was investigated the effect of month of age at the first pregnancy on calving difficulty of Holstein heifers. Heifers was inseminated with semen of easy-calving sire and the reproductive performance was compared for the effect of first pregnancy at 12-14 months of age (12 month old), 15-16 months of age (15 month old), 17-19 months of age (17 month old) and > 20 months of age (20 month old) in Holstein heifers. Holstein heifers showed different easy calving score (ECS) depending on their ages; frequency of ECS 1 was 55.2%, 64.9%, 59.4% and 52.0% at 12, 15, 17 and 20 month of age, respectively. Frequency of ECS 4 was 5.2%, 1.8%, 6.3% and 4.8% at 12, 15, 17 and 20 month of age, respectively. The rates of retained placenta postparturition in first pregnancy age category was 12.3% at 12 months of age, 5.3% at 15 months of age, 9.4% at 17 months of age and 19.0% at 20 months of age. When It was evaluated by present results, the reproductive performance like easy calving score (ECS), retained placenta rates, and abortion and stillbirth rates of heifers which was inseminated at 14 months old or less and at 17 months old or more was reduced. Therefore, the first pregnancy age for optimum reproductive performance was 15-16 months.

This study suggests designs of maternity clothes wearable after childbirth and aims at confirming that maternity wear is continuously wearable during a post-natal period. As for research methods, theoretical and empirical studies with research on a production of work were conducted, focusing on the design development of the maternity dress which may be worn even after childbirth. The results of theoretical and empirical studies possibly presented various designs which require fashionability going with the trend of time. The maternity wear also requires the adaptability for a changing body shape to enable a woman to wear it during a pregnancy period and after childbirth. Besides, the convenience of dressing and undressing with wearing comfort is considered. 6 sets of one-piece dress were made as the result of the study and three factors of fashionability, adaptability and convenience were considered. The conclusions are as follows : Firstly, it verified that the maternity wear is fashionable clothes not to cover protruded abdomen but to satisfy women's desire of self expression. Secondly, it overcame the limit of dress design through fitting it to completely different two body sizes and shapes with design factors and patterns. Lastly, it suggested a research model of maternity wear corresponding to economical, environmental and ethical principles as giving an effect of “two” dresses with one. This study presented a new direction of design study with meaningful actions of fashion designing in the aspects of economical, environmental and ethical consumption unlike artificial disposal on SPA brands.