This study was carried out to produce superior dairy cattle by embryo transfer. Seven dairy cows were superovulated with divided injection of FSH 4Omg for 5 days started on day 9 to 14 of the estrus cycle and injection of PGF$_2$$\alpha$ 45mg on day 4 of FSH injection. Donor cows were flushed to collect embryos on day 7 or 8 of the estrus cycle. Fresh embryos collected were transferred to synchronized dairy recipients or frozen using glycerol 3 step method to he equilibrated. And 35 embryos which were frozen using glycerol 6 step method were imported from U.S.A. After glycerol dilution of frozen embryos was done by reverse density during freezing. frozen-thawed embryos were transferred to synchronized dairy or beef recipients. The results obtained were as follows; 1. Total of 24 embryos were collected from 7 donor cows flushed and transferable embryos were 18 (75.0%). 2. Among 24 embryos. morula, early blastocyst, blastocyst, expanded blastocyst and unfertilized ova were 3 (12.5%), 1 (4.2%), 10 (41.6%), 4 (16.7%) and 6 (25.0%), respectively. 3. Heat inducing rate after 1st and 2nd injections of PGF$_2$$\alpha$ in Holstein and beef cattle was 83.3% and 71.4% and 62.5% and 69.2%, respectively. 4. Among 56 recipients, 23 head were pregnant (41.1%). The pregnancy rate of fresh embryos was 50.0% (1/2 heads) and the pregnancy rate of frozen embryos which were frozen using glycerol 3 step and using glycerol 6 step imported from U.S.A. was 52.6%(l0/19 heads) and 34.3%(12/35 heads), respectively. 5. The pregnancy rate of blastocyst (60.0%) was higher than that of morula (39.0%), early blastocyst (25.0%) and expanded blastocyst (0%). 6. The pregnancy rate of grade I embryos (52.2%) was higher than that of grade 2 (34.6%) and grade 3 (28.6%). 7. The pregnancy rate according to synchrony of recipient with donor was higher in simultaneous recipient (55.0%) and +l2hrs' (53.8%) than -24hrs' (23.5%), -l2hrs' (20.0%) and +24hrs' (0%).
The purpose of the present study was to produce live offspring from in vitro fertilized goat embryos. Oocytes were collected from abattoir ovaries and kept in oocyte collection medium. Oocytes were washed 4-5 times with maturation medium containing medium-199 with 5 ${\mu}g/ml$ FSH, 100 ${\mu}g/ml$ LH, 1 ${\mu}g/ml$ estradiol-$17{\beta}$ 50 ${\mu}g/ml$ gentamycin, 10% inactivated estrus goat serum, and 3% BSA (fatty acid free). Oocytes were placed in 100 ${\mu}l$ drops of maturation medium containing granulosa cell monolayer and incubated in a 5% $CO_2$ incubator at $38.5^{\circ}C$ for 27 h. For capacitation of spermatozoa fresh semen was processed and mixed in 3 ml fertilization TALP medium containing 50 ${\mu}g/ml$ heparin and kept in the above incubator for 2 h. The capacitated spermatozoa were coincubated with matured oocytes for fertilization. Cleaved embryos were separated and cultured in embryo development medium with oviductal cells and 494 embryos were produced. Recipient goats were synchronized with two injections of 15 mg $PGF_{{2}{\alpha}}$/goat 10 days apart. Eighty early stage embryos were transferred into the uterotubal junction of 14 surrogate mothers using laparoscopy techniques. One recipient delivered twin kids, whereas another two recipients each.delivered a single kid The parentage of these kids was evaluated using highly polymorphic co-dominant microsatellites markers. From the present study, it was concluded that live goat kids can be produced from in vitro matured and fertilized goat embryos, to the best of our knowledge for the first time in India.
Kim, Ill-hwa;Son, Dong-soo;Lee, Kwang-won;Chang, In-ho
Korean Journal of Veterinary Research
/
v.32
no.1
/
pp.143-151
/
1992
Sixty Four fresh and 142 frozen embryos of dairy cattle were transferred to synchronized dairy, beef or Korean Native Cattle nonsurgically at National Animal Breeding Institute from 1985 to 1990. The results obtained were as follows ; 1. The pregnancy rate of fresh embryos(39.1%) was higher than that of frozen embryos(32.4%) and average pregnancy rate was 34.5%. 2. The pregnancy rate of grade 1 embryos was higher than that of grade 2 embryos for both fresh(41.3% vs 33.3%) and frozen embryos(35.4% vs 25.6%). 3. The pregnancy rate according to development stage of fresh embryos was increased with maturity as 29.2%, 33.3%, 50.0% and 54.5% for morula, early blastocyst, blastocyst and expanded blastocyst, respectively. For frozen embryos, the pregnancy rate of blastocyst(44.4%) was higher than those of morula(31.3%) and early blastocyst(28.0%). 4. The pregnancy rate according go recipient-donor synchrony for fresh embryos was higher when the recipients exhibited estrus 1 day earlier than the donors(43.8%) than when the recipients exhibited estrus 1 day later than the donors(38.1%) or when the recipients and donors exhibited estrus at the same time(37.0%). For forzen embryos, the pregnancy rate was decreased when the recipients and donors exhibited estrus at the same time(37.9%), when the recipients exhibited estrus 1 day later than the donors(32.0%) and when the recipients exhibited estrus 1 day earlier than the donors(23.5%), in sequence. 5. The pregnancy rate of heifers was higher than that of cows for both fresh(50.5% vs 37.9%) and frozen embryos(39.7% vs 25.7%). 6. The pregnancy rate according to recipient breed for fresh embryos was higher in dairy cattle(42.1%) and beef cattle(40.%) than in Korean Native Cattle(33.3%). For frozen embryos, the pregnancy rate was decreased beef cattle(39.1%), dairy cattle(30.3%) and Korean Native Cattle(14.3%), in sequence. 7. The pregnancy rate according to equilibrium steps of glycerol and freezing rate was higher when transferred after 3-steps equilibrium and freezing by the rate of $0.3^{\circ}C$/min from $-6^{\circ}C$ to $-35^{\circ}C$ and $0.1^{\circ}C$/min to $-38^{\circ}C$(39.4%) than when transferred after 6-steps equilibrium and freezing by the rate of $0.5^{\circ}C/min$ from $-6^{\circ}C$ to $-30^{\circ}C$(30.3%).
Cloning technology continues to capture widespread attention by the international news media and biomedical and agricultural industries. The future uses of this technology could potentially contribute to major advances in biomedical and agricultural sciences. Cloned transgenic dairy cattle possessing milk promoters directing transgenes will produce pharmaceutical proteins in their milk faster, more efficiently and less expensively than transgenic cattle created using microinjection techniques. Additionally, cloned transgenic fetuses and animals may become a source of cells, tissue and organs for xenotransplantation. Lastly, but maybe most importantly, enhanced production traits and disease resistance may be realized in animal agriculture by utilizing these new technologies. The recent advances in the cattle cloning technology are important but there are still major obstacles preventing widespread commercial use of this technology. The type of donor nucleus, recipient cytoplasm, and cloning procedures used will impact the potential number of clones produced and the uses of the technology. In addition, the new advances in cloning methodology have not improved the relatively low pregnancy rates or reduced the incidence of health problems observed in cloned offspring. These problems may require novel techniques to decipher their cause and new methods of preventing and/or diagnosing them in the preimplantation embryo. The commercial potential is enormous for cloning technology; however, little has been done to improve the efficiencies of the procedure. Improving procedural efficiencies is a critical developmental milestone especially for potential uses of cloning technology in animal agriculture.
This study evaluated the effect of recipient's breed on the gestation length and birth weight of Hanwoo calves produced by Hanwoo ovum pick up (OPU) derived embryos. Embryos of OPU derived Hanwoo cows were transferred into the Hanwoo and Holstein recipients at 6~7 days of estrus cycle. Gestation length was expressed in days from the day of embryo transfer to the birth day of offspring, and birth weight of offspring was recorded within 24 h of birth. Breed of recipient cows (Hanwoo and Holstein) has no effect on overall gestation length ($280.9{\pm}6.2$ vs. $284.4{\pm}9.8$ days) and birth weight of calves ($23.56{\pm}3.75$ vs. $27.70{\pm}7.92\;kg$). The gestation length was higher (p<0.05) for male calves than female calves in both Hanwoo ($283.8{\pm}6.7$ vs. $277.3{\pm}3.3$ days) and Holstein ($287.3{\pm}8.9$ vs. $280.1{\pm}9.1$ days) recipient cows. Birth weight of Hanwoo calves did not differ when Hanwoo recipients ($26.5{\pm}1.3$ vs. $23.3{\pm}1.2\;kg$) were used for embryo transfer. However, male calves were heavier (p<0.05) at birth than that of female calves when embryos were transferred into the Holstein cow ($33.5{\pm}4.9$ vs. $27.8{\pm}4.9\;kg$). In conclusion, the present results demonstrated that breed of recipient cows have no effect on gestation length and birth weight of OPU derived Hanwoo calves. However, gestation length and birth weight of male calves were higher in both Hanwoo and Holstein recipient cows.
The purposes of this study were to: (a) investigate the characteristics of recipients' of the non-government foodbank program, (b) examine the health and dietary related conditions of them, and (c) evaluate the benefits and effectiveness of the foodbank program from the recipients' perspective. A total of 21 groups (n = 755) and 75 individual recipients participated in the survey. The main results of the study were as follows; (a) Generally, the individual recipients were 74-year-old female, livelihood protectee, and those who received government assistance or funds from private donators as their source of livelihood. (b) The ages of group recipients varied widely, and they also received government assistance or funds from private donators as their source of livelihood. (c) Most of the donated foods were bakery and confectionery Items. rice, and milk and other dairy products. (d) Benefits such as the decrease in the recipients' food expenses and an enhancement of their nutritional statuses were identified. (Korean J Community Nutrition 8(2) : 231∼239, 2003)
The development of flip-graft, yield of antler and growth of antler in Sika deer, Red deer and Elk were investigated to establish the technology of transplantation of flip-graft. Pedicle flip-grafts were transplanted on the sides pedicle, on the midline of forehead and on the middle line of the skull. The success rate of transplantation was 42%, 5 animals out of 12. The yields of antler and number of point were 716g and 1,071g, and 2.3 and 1.0 in control and donor of flip-graft, respectively in red deer. The length and weight of anther of recipient of flip-graft in elk were 17cm, 302g. The length and weight of hard antler of recipient of flip-graft in red deer were 26cm and 122g. Flip-graft should be transplanted on the scull after removal of the corium for strong adherence.
In order to study the changes of pregnancy rates according to the factors, age, season, repeated utilization, synchrony, of recipients (29 holstein heifers, one Korean native heifer and 3 holstein cows) were transfered one embryo (from morulae stage to advanced blastocyst) to each, and the results are as follows ; 1. The effect of age of recipient on pregnancy rate in cows and 18-24months of aged group are 100% and 78%, respectively. Under 14months of aged group recorded the worst, 67%. 2. In winter (Nov.-Jan.) and spring (Feb.-Apr.), the pregnancy rate were better than others 100%, 83%, respectively. In the autumn (Aug.-Oct.), it was only 50%, the worst. 3. Among 31 recipients which were utilized at the first, 25 were pregnanted (80.7%), but in the second utilization after the failure of the first transfering, 2 heifers, none of pregnancy was obtained. 4. Pregnancy rates related to synchronization of estrus between donor and recipients are;-(before donor)12hrs. (100%), -6hrs. (86%), +( after donor)6hrs. (67%), + 12hrs. (79%), and +over 12hrs. (50%), respectively. 5. Differences in the pregnancy rate of the recipients +over 12hrs. were significantly large, so in order to ger good pregnancy rate in embryo transfer, the differences of the synchronization must be reduced inner ${\pm}12hrs$. In general, recipients estrus before donor showed better pregnancy rate than estrus after donor.
Park, Sang-Hoon;Lee, Mi-Ran;Kim, Tae-Suk;Baek, Sang-Ki;Jin, Sang-Jin;Kim, Jin-Wook;Jeon, Sang-Gon;Yoon, Ho-Baek;Lee, Joon-Hee
Reproductive and Developmental Biology
/
v.38
no.4
/
pp.147-158
/
2014
Differentiated nuclei can experimentally be returned to an undifferentiated embryonic status after nuclear transfer (NT) to unfertilized metaphase II (MII) oocytes. Nuclear reprogramming is triggered immediately after somatic cell nucleus transfer (SCNT) into recipient cytoplasm and this period is regarded as a key stage for optimizing reprogramming. In a recent study (Dai et al., 2010), use of m-carboxycinnamic acid bishydroxamide (CBHA) as a histone deacetylase inhibitor during the in vitro early culture of murine cloned embryos modifies the acetylation status of somatic nuclei and increases the developmental competence of SCNT embryos. Thus, we examined the effects of CBHA treatment on the in vitro preimplantation development of porcine SCNT embryos and on the acetylated status of histone H3K9 on cloned embryos at the zygote stage. We performed the three groups SCNT: SCNT (NT), CBHA treatment at the porcine fetus fibroblast cells (PFFs) used as donor cells prior to SCNT (CBHA-C) and CBHA treatment at the porcine SCNT embryos during the in vitro early culture after oocyte activation (CBHA-Z). The PFFs were treated with a $15{\mu}M$ of CBHA (8 h) for the early culture and the porcine cloned embryos were treated with a $100{\mu}M$ concentration of CBHA during the in vitro early culture (10 h). Cleavage rates and development to the blastocyst stage were assessed. No significant difference was observed the cleavage rate among the groups (82.6%, 76.4% and 82.2%, respectively). However, the development competence to the blastocyst stage was significantly increased in CBHA-Z embryos (22.7%) as compared to SCNT and CBHA-C embryos (8.6% and 4.1%)(p<0.05). Total cell numbers and viable cell numbers at the blastocyst stage of porcine SCNT embryos were increased in CBHA-Z embryos as compared to those in CBHA-C embryos (p<0.05). Signal level of histone acetylation (H3K9ac) at the zygote stage of SCNT was increased in CBHA-Z embryos as compared to SCNT and CBHA-C embryos. The results of the present study suggested that treatment with CBHA during the in vitro early culture (10 h) had significantly increased the developmental competence and histone acetylation level at the zygote stage.
This study was conducted to investigate the effect of embryo stage on bisection rates of embryos, on development of separated demi-embryos and on subsequent development to full term following transfer of demi-embryos to recipients. The results obtained in this study are summarized as follows. The 2-, 4-, 8-cell and morula embryos were mostly obtained at 36-42, 48-54, 62-66 and 72-78 hours after injection of hCG, respectively, and the average number of embryos recovered per head was 17.0. The bisection rate of 8-cell embryos was 83.6%, which was significantly higher than that of morula embryos, 65.5%. But the development of morula demi-embryos to blastocyst after in-vitro culture was better than that of 8-cell demi-embryos and the rate was 76.5% and 60.9%, respectively. When the 2, 10 and 15 cultured demi-embryos or intact embryos were transferred to each recipient mouse, respectively. the highest pregnancy rate was obtained when 15 embryos were transferred. And the overall offspring production rate of intact embryos was higher than that of demi-embryos.
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