• Asian-Australasian Journal of Animal Sciences
• /
• v.1 no.3
• /
• pp.153-156
• /
• 1988

Bovine blastocysts were obtained by in-vitro culture of embryos derived from in-vitro fertilization of oocytes matured in-vitro. These blastocysts and blastocysts from inseminated donors were bisected by a simple method (without a holding pipette) using a microblade operated by a micromanipulator. A pair of demi-embryos was transferred nonsurgically into each uterine horn of a recipient cow 6 or 8 days after estrus. Pregnancy resulted from the third transfer. Ultrasound examination done 52 days after estrus (46 days after transfer) confirmed the presence of at least one fetus in the each uterine horn. This is the first report to show the viability of bisected bovine blastocysts obtained from in-vitro culture of embryos derived from in-vitro fertilization of oocytes matured in-vitro. In addition, a simple method to bisect bovine embryos is described.

• Journal of Embryo Transfer
• /
• v.14 no.2
• /
• pp.93-97
• /
• 1999

The development potential of bovine somatic cells was evaluated using nuclear transfer. A single donor cell derived from fetus of HanWoo(Korean Native Cattle) was selected and deposited into perivitelline space of each enucleated oocyte before electrical fusion and activation. Nuclei of donor cells starved for 7 days (37%) tended to support the development of reconstitute embryo the blastocyst stage better than those of donor cells starved 3, 14 and 30 days. The cleavage rate was significantly lower(P<0.05) in reconstitute embryos derived from large size donor cells(51.2%), than those from small medium size donor cells(76.6 and 73.5, respectively). The developmental rate to blastocyst of reconstructed embryos from medium size donor cells was higher than those from small and medium size donor cells. This study demonstrates that an appropriate culture period for induction into quiescent stage and the size of donor cells effect on the efficiency of nuclear transfer using cultured bovine cells.

• Proceedings of the Korean Society of Developmental Biology Conference
• /
• 2003.10a
• /
• pp.140-140
• /
• 2003

The study evaluated the effect of donor cell treatments for G0/Gl synchronization and the donor ceil type on development and incidence of apoptosis in cloned cattle embryos. Primary cultures were established from a female fetus on day 50 of gestation and adult ear skin biopsies. Cells were randomly allocated into 3 experimental treatment groups after 6~8 passages. Group 1 (Confluent), cells were cultured in DMEM supplemented with 10% FBS until 90% confluent. Group 2 (Serum-starvation), cells were cultured in DMEM Supplemented With 0.5% FBS for 5 days. Group 3 (Roscovitine), Cells were cultured in DMEM supplemented with 10% FBS and 30 $\mu$M Roscovitine for 12 h. Cell cycle and apoptosis were analyzed using flow cytometry after labelling with DAPI and YO-PRO-1. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and fused by a single DC pulse (1.6 kV/cm, 60 $\mu$sec). (중략)

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.3
• /
• pp.346-351
• /
• 2002

The effect of fetal number (single or twin) on plasma concentrations of progesterone, estradiol $17{\beta}$, cortisol, prolactin, growth hormone, triiodothyronine, thyroxine and insulin around parturition (periparturient period) were studied on ten $Alpine{\times}Beetle$ crossbred goats in their first to third lactation. The hormone profiles were studied on days -20, -15, -10, -5, -4, -3, -2, -1 prior to kidding and on day 0 and +1, +2, +3, +4, +5, +10, +15, +20 days postkidding. Plasma progesterone levels were significantly (p<0.01) higher in twin bearing goats comparison to single bearing goats during all the days of sampling. The decline in progesterone concentration from day 20 to day 1 before kidding was 56% in twin and 42% in single bearing goats. In single bearing goats plasma estradiol $17{\beta}$ was significantly (p<0.01) higher during prekidding days compared to twin bearing goats. The level of estradiol $17{\beta}$ was highest on the day of kidding in both the groups. The plasma prolactin level in twin bearing goats from day 10 to day 1 prepartum was higher as compared to single fetus bearing goats. However there was abrupt increase in prolactin level on the day of kidding in both the groups. The plasma growth hormone levels were significantly (p<0.01) higher in twin compared to single bearing goats. On the day of kidding growth hormone levels were significantly (p<0.01) higher in twin as compared to single bearing goats (1.40 vs. 0.95 ng/ml). In twin bearing goats plasma cortisol values from day 5 till the day of kidding remained elevated and the levels on the day of kidding was significantly highest in both the groups. The levels of triiodothyronine ($T_3$) were significantly higher (p<0.01) during all the periods of sampling in single compared to twin bearing goats. Plasma thyroxine ($T_4$) was significantly (p<0.01) lower in twin compared to single bearing goats. In single bearing goats plasma insulin levels were significantly (p<0.01) higher than twin bearing goats during prepartum period however during post partum period the levels in both the groups remained similar. It can be concluded that number of fetuses is having significant influence on the hormone profile during periparturient period.

• Journal of Embryo Transfer
• /
• v.25 no.1
• /
• pp.43-50
• /
• 2010

The specimens from 32 aborted fetus and 274 aborted cows were collected in 168 farms of Chonnam province from 2005 to 2008 and were tested the brucellosis. The results obtained are summarized as follows. In the 32 aborted fetus, bovine brucellosis was detected in 12 heads (37.5%), bovine viral diarrhea-mucosal disease was detected in 7 heads (19%), ainovirus infection was detected in 1 head (3.1%), and multi-infection of BVD and brucellosis was not detected, respectively. In the 306 cases of aborted fetus and cows, bovine brucellosis was detected in 44 heads (14%). Status of abortion were confirmed in 63 farms (38%) out of 168 farms from June to August. From the point of raising scale, studies found that 128 farms (76%) out of all raised under 20 heads. The incidence of abortion by brucellosis was mainly showed in 30 heads (68.1%) about 151~250 days of gestation. In the result of the 18 farms survey, the causes of infections were detected movement of infected cattle in 5 farms (28%), unknown cause in 12 farms (67%), and recurrence in 1 farm (5%). The results of this study suggest to take an advantage of the prevention and fundamental research for bovine brucellosis in Chonnam province.

• Journal of Animal Science and Technology
• /
• v.65 no.4
• /
• pp.818-837
• /
• 2023

Understanding adipocyte development in fetus during bovine pregnancy is important for strengthening fattening technology. Additionally, nutritional level of dams during pregnancy has the potential to improve offspring growth and fat development. The purpose of this study is to evaluate the intramuscular adipocyte development and expression level of related genes in bovine fetus, and the effect of increased crude protein (CP) intake during pregnancy on the growth performance and carcass characteristics of male offspring. Eighty six pregnant Hanwoo cows (average body weight, 551.5 ± 51.3 kg, age 5.29 ± 0.61 y) were used. Fetuses were collected at 90, 180 and 270 d of gestation from 18 pregnant Hanwoo cows. The remaining 68 pregnant cows were randomly assigned to 2 feeding groups. The control (CON) group was provided the standard protein diet (n = 34), and treatment (TRT) group was provided a diet with a 5% increase in CP intake (n = 34). Male offspring were divided into two groups according to protein treatment of the pregnant cows: CON male offspring (CON-O) and TRT male offspring (TRT-O). Intramuscular adipocytes were found in the fetal skeletal muscle after 180 days of gestation. Male calf's birth weight increased in the TRT group compared to that in the CON group (p < 0.002). The final body weight (p < 0.003) and average daily gain (p < 0.019) of male offspring were significantly higher in TRT-O than in CON-O. The feed conversion ratio was also improved by 10.5% in TRT-O compared to that in CON-O (p < 0.026). Carcass weight was significantly higher in the TRT-O group than that in the CON-O group (p < 0.003), and back fat was thicker in the TRT-O group (p = 0.07). The gross receipts and net income were higher in TRT-O than in CON-O (p < 0.04). Thus, fetal intramuscular fat can be formed from the mid-gestation period, and increased CP intake during pregnancy can increase net income by improving the growth and carcass weight of male offspring rather than intramuscular fat.

• Journal of Embryo Transfer
• /
• v.30 no.3
• /
• pp.219-224
• /
• 2015

The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.

• Korean Journal of Veterinary Research
• /
• v.40 no.1
• /
• pp.42-48
• /
• 2000

Akabane disease is transmitted through mosquitoes in cattle, sheep and goats. It shows congenital abnormalities including encephalomyetitis, hydranencephaly, neurogenic arthrogryposis, and deformed neonatal calves. Akabane viruses, 93FMX and K-9 strain, were isolated from fetal matrix of aborted cow and blood of healthy cow, respectively. S gene sequences of 93FMX and K-9 showed 100% homology with that of OBE-1 strain isolated in Japan. Based upon our sequencing data, we synthesized specific primers for PCR diagnosis. Using these primers, we were able to amplify the S gene of Akabane virus not only from the culture fluid of Vero cells but also from the brain tissue of suckling mouse inoculated with, Akabane virus. These PCR products were confirmed by Southern blot hybridization. Not only the sensitivity of PCR test was high enough to detect the viruses of $10^{1.0}TCID_{50}/ml$, but also the time for diagnosis was significantly shorter than that of the virus isolation by tissue culture method. This method was also effective for the detection of Akabane virus in the cerebrum of fetus. RT-PCR method may be used for a useful diagnostic test of the clinical cases of Akabane disease.

• Journal of Embryo Transfer
• /
• v.28 no.4
• /
• pp.343-348
• /
• 2013

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

• Journal of agriculture & life science
• /
• v.44 no.5
• /
• pp.65-74
• /
• 2010

Dairy cow undergoes major adaptations in glucose and lipid metabolism to meet fetal and mammary nutrient requirements during the periparturient period. These adaptations are characterized by major changes in response to the homeostatic signal of insulin. In response to insulin, fatty acids are mobilized to meet energy demand. And cow develop insulin resistance during the late prepartum period to facilitate glucose sparing for the fetus and mammary gland. Insulin resistance is an important adaptation and may be partly responsible for the large increase in plasma nonesterified fatty acid (NEFA) that occurs prior to parturition and coincides with the characteristic decrease in dry matter intake (DMI). Modulation of insulin resistance by administration of thiazolidinedione (TZD) during the periparturient period, likely through the activation of $PPAR{\gamma}$, has substantial potential to minimize energy and immune-related metabolic disorders, and thus may result in increased productivity and improved health of dairy cows.