This study was carried out to establish an effective embryo production system by analyzing several factors ifluencing in vivo embryo production in Hanwoo. The results are summarized as follows : 1. The number of total, fertilized and transferble embryos by gonadotropins were no significant difference among PMSG, FSH and FSH+PMSH. 2. The numbers of recovered and transferable embryos by type of FSH were no significant difference in both FSH-P and FOLLTROPIN-V but significant difference with SUPER-OV(p<0.01). 3. The numbers of recovered and transferable embryos by age of donor were highest in 5 years old. 4. The recovered number of transferable embryos was highest in the cows at the 1 st and 3rd parity but significantly lower in hifers and the cows at more than 6th parities(p<0.01). 5. The numbers of trasferable embryos by postparient were more<270 days than > 270 days in significant difference(p<0.01). 6. Embryo production after treatment of repeated super ovulation was no significant difference. 7. Recovered and trasferable embryos production after treatment of repeated super ovulation interval was highest 81∼100 days and significant difference(p<0.01). 8. The numbers of recovered and transferable embryos by seasons were no significant difference.
The objective of this study was to investigate the comparison of transferable embryos and pregnancy rate between Hanwoo and Chickso. The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on Hanwoo 78%, and Chickso 85%, respectively. The mean number of total embryos are each $14.76{\pm}2.16$ and $6.23{\pm}1.07$. So the mean number of transferable embryos are each $10.94{\pm}1.91$ and $4.58{\pm}1.05$. In addition, the mean number of total Hanwoo embryo from <10 and $10{\leq}$ of corpora luteum was $0.50{\pm}0.50$, $11.56{\pm}1.92$, respectively. In case of Chickso, The mean number of transferable embryo from <10 and $10{\leq}$ of CL was $2.75{\pm}1.39$, $6.00{\pm}1.00$, respectively. The pregnancy rates were Hanwoo 40%, and Chickso 37% following transfer of fresh embryos produced in vivo. Also, the pregnancy rates of Chickso 60% were significantly greater (p<0.05) than the Hanwoo 42.48% following transfer of following transfer of frozen embryos, respectively. In conclusion, these results suggest that Chickso may be effectively used for transferable embryos production in Hanwoo. Although the transferable embryos number was not enough, it seems the Chickso greatly affect pregnancy rate. The results indicated that the possibility of transferable embryos from Chickso for embryo transfer could be confirmed in this study. Based on the present findings, it was suggested that it is very important to evaluate in vivo embryo production and pregnancy rate of embryo transfer following superovulation for effective Hanwoo and Chickso production.
This study was performed in order to simplify the operation and minimize stress of donor and be readily available in the field with low cost and high quality embryos using the Direct Embryo Collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3rd day administration of FSH, 25 mg $PGF_2{\alpha}$ was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1 st insemination and embryos were recovered 8 days after the 1st insemination. Embryo collection from superovulated donors was performed to flushing by non-surgical methods of 3-way, 2-way and DEC (l-way). The average number of recovered embryos were 11.25${\pm}$0.63, 12.5${\pm}$0.65 and 11.75${\pm}$0.48 from operations of 3-way, 2-way and DEC methods, respectively. There were no significant differences among the embryo collection methods. Also, The average number of transferable embryos were 6.25${\pm}$0.48, 7.25${\pm}$0.48 and 7.25${\pm}$0.63 from each embryo collection procedures. The number of transferable embryos was no differences among the 3-way, 2-way and DEC methods, respectively. Meanwhile, the ratio of transferable embryos for all recovered embryos from DEC methods was higher as 61.7 % than 55.6 %, 58 % from methods of 3-way, 2-way. And the flushing solution required for recovering embryos by DEC method was significantly lower as 0.28${\pm}$0.32 1 than 1.8${\pm}$0.12 1, 1.75${\pm}$0.10 1 from 3-way, 2-way methods (p<0.05). Also, the time required for recovering embryos by DEC methods was significantly lower as 27${\pm}$2 min than 51${\pm}$3, 45${\pm}$2 min, respectively (p<0.05). In conclusion, these results suggest that DEC method for embryo collection may be effectively used for production of in vivo embryos using less flushing solution and, it might be effectively available in the field compared to conventional embryo recovery methods using 3-way or 2-way balloon catheter.
This study was carried out to establish an effective system for embryo transfer techniques by analyzing several factors affecting in-vivo embryo transfer in Korean cattle. Embryos produced in-vivo were transferred into a total of 301 recipients. The results obtained in studies on the factors affecting pregnancy rate after embryo transfer by condition of embryos were as follow ; 1. The pregnancy rate of 301 recipients was 45.2% and higher with fresh embryos than with frozen embryos(63.5% : 21.4%, P<0.01). Embryos superovurated by FSH-P had slightly greater than by SUPER-OV in pragnancy rate, athough these were no difference between two treatments. 2. The pregnancy rates of transferred morulae and blastocysts showed no difference between fresh and frozen embryos(63.5% : 63~6% ; 20.0% : 25.8%). However, the pregnancy rates by quality of flesh and frozen embryos were significantly different(P<0~05). The pregnancy rates were outstandingly high in the grade A, B of fresh embryos(59.0~66.4%), and in the grade A of frozen embryos(43.6%). 3. The number of transferred embryos showed no difference in pregnancy rate, but when frozen embryos transferred, the pregnancy rate was slightly higher with two embryos than that with one embryo.
An ecological investigation was conducted in winter from November 2007 to February 2008, around Wangdol Reef in the East Sea to analyze the population structure and fecundity of Chinoecetes opilio. The sex ratio was 30.11:69.89% (female:male) in November, 62.03:37.97% in December, 37.59:62.41% in January and 14.69:85.31% in February. Regression equations indicated highly positive relationships between size and weight parameters (carapace length, width, total weight). During the sampling period ovigerous females were continuously collected. The percentage of ovigerous females was 96% in November, 90.57% in December 89.80% in January, and 88.46% in February. The average number of embryos was 64,800. The regression equation between carapace width and the number of embryos was y = 2.6805x - 0.9182 ($R^2$ = 0.7166). Embryo volume increased as embryo development proceeded. The mean size of an embryo was 0.72 mm. Embryo volume ranged from 0.42 to $0.84\;m^3$ during embryo developmental stages 1 to 3.
To investigate the usability of frozen canine embryos for embryo transfer in the dog, 19 donors, 3 recipients, and 6 male dogs were used for the experiment. Natural mating or artificial insemination was performed for breeding the bitches in natural estrus. Vaginal smear test along with progesterone titre test were performed to detect the appropriate mating time and the bitches were bred twice during 3-6days following LH surge. Embryo collection was done on 8, 9-11, 12-13 days after the second mating to collect morula and blastocyst. Embryos were frozen using a programmable freezer and preseued in LNE tank. Embryos were thawed in 37$^{\circ}C$ water for 15 seconds and transferred into each uterine horn within 30 minutes. Embryos were collected from 13 bitches of 19 donors(68.4%) and the collected embryos were from between 9 and 13 days after 2nd mating. Embryos were produced both by natural mating(60.0%, 9115) and AI with frozen semen(100.0%, 4/4). Embryos were collected from the donors weighed between 2.5 and 30 kg and their age was from 1.5 to 3 years. 52 embryos were collected from 13 donors and the mean number of embryos was four. The stage of embryos was from 2-cell to gastrula and morulae were colledted mostly from 10 to 11 days after 2nd mating. Embryos were collected evenly from each uterine horn and the rate of embryo collection for the number of corpus luteum was 83.9%. Embryos were transferred to 3 recipients(morula 8, blastocyst 1, gastrula 8), however, no offspring was produced.
Embryos formed in vivo were collected from 171 donors housed in Chung Cheong Buk-Do Institute of Livestock and Veterinary Research of the Chungbuk community during the years 2009~2012. We evaluated annual embryo collection, effect of follicle stimulating hormone (FSH), controlled internal drug release (CIDR) and prostaglandin (PG) administration to the donor for superovulation and controlling the estrus cycle, seasonal effects of embryo collection and compared the number of embryos recovered as per the collection days and pregnancy rate. In all, 1,243 embryos were collected from 118 donors with an average of $7.31{\pm}5.35$ embryos per donor, out of which 69.4% were transferable. Dosages of FSH required for inducing superovulation in various donors were compared. Average number of embryos collected from donors administered with 30 AU of FSH ($7.13{\pm}5.74$ per donor) was not significantly different from that of donors who were given an injection of 24 AU of FSH ($7.53{\pm}4.91$ per donor). However, the percentage of transferable embryos in the 30AU FSH-administered group (63.2 %, 449 of 711) was higher than that in the 24AU FSH-administered group (77.8%, 414 of 532). In the group of donors under a natural estrus cycle, the FSH dose administered did not influence the number of transferable embryos produced ($7.49{\pm}6.25$ per donor for 30 AU of FSH vs $7.49{\pm}4.92$ per donor for 24 AU of FSH). However, in donors administered with CIDR and PG for controlling the estrus cycle, the FSH dose affected the average number of transferable embryos collected ($4.25{\pm}2.87$ per donor for 30 AU of FSH vs $8.50{\pm}6.36$ per donor for 24 AU of FSH). We collected embryos from donors 6, 7 or 8 days after artificial insemination (AI). Results showed that the percentage of transferable embryos among those collected 8 days after AI was significantly higher than that among embryos collected 6 or 7 days after AI. Seasonal variations did not affect number of recovered embryos and pregnancy rates in natural estrus cycle and CIDR treatment groups (48.28% and 42.55%) but higher than pregnancy rate of frozen embryos (19.63%). These results indicated that administration of FSH beyond a threshold dose (at least 24 AU) has no beneficial effect on the production embryos and that collection of embryos 7~8 days after AI is optimal for embryo recovery. CIDR treatment induced superovulation in short term and had no influence on the natural estrus cycle. Finally, although good-quality embryos were transferred, freezing significantly reduced the pregnancy rates after transfer.
The possibility of embryo transfer technique in pigs to introduce a new genetic material into closed herds for disease control was investigated. The results investigated were as follows: 1. The exhibiting rate of estrus on the administration of altrenogest and PMSG ranged from 83.3 to 100% and the estrus exhibited within 4.0 - 5.3 days after the administration of altrenogest. 2. The average number of ovulation points per pig by the injection of PMSG and HCG were 14.0 - 30.7. 3. The average number of recovered embryos per pig was 15.7, and 72.8% of embryos were recovered. 4. The pregnancy rate of recipients was 63.6% and the survival rate of transferred embryos were 21%. 5. When the 1-cell embryos were cultured for 24hrs, they were all developed. Therefore it is possible to recover and preserve the embryos from the donor pigs which have high genetic ability, and to transfer embryos to recipient pigs which are separated from donor pigs in Korea.
Fine wool sheep (n=18) maintained in a tropical environment were allocated to three treatment groups. Estrus was induced with two injections of $PGF_{2{\alpha}}$ (10 mg. im) at 10 days interval. Superovulation treatment started 2 days prior to the second injection of $PGF_{2{\alpha}}$. Each ewe was treated with a total dose of 25 units FSH (Super-OV) i.m. every 12 hover 3 days; Group 2 were also injected i.m. with 200 IU PMSG at the first injection of FSH; Group 3 was treated as in Group 2 and also with GnRH ($4{\mu}g$ Buserelin) at the onset of estrus. The ewes in estrus were mated with a fertile ram. Ovarian examination and recovery of embryo and ova were performed at laparoscopy and laparotomy on day 3 or 4 after mating. Data for onset of estrus, duration of estrus, number of corpora lutea (CL), number of unnovulated large follicle (LF), embryo recovery rate, embryo quality and fertilization recorded for the 3 groups. Ewes in the Group 1 set in estrus later (p<0.05; $50.0{\pm}7.29h$) than the ewes in Group 2 ($24.5{\pm}3.58$) and 3 ($32.5{\pm}3.58h$). The duration of estrus, ovarian size and ovarian response (number of CL and LF) did not differ significantly (p>0.05) among the 3 groups. The proportion of ewes with a superovulatory response (${\geq}2$ CL) was the lowest (50%) in Group 1 treated with FSH alone but ova/embryo recovery (100%) and fertilization (100%) was significantly (p<0.05) higher than Group 2 (58.3 and 85.7%, respectively) and Group 3 (48.6 and 50%, respectively). It is concluded that in tropical fine wool sheep, there is no difference in the 3 treatments for yield of good quality embryos but ovarian response and ovulation rate increased on additional use of PMSG and GnRH respectively to FSH alone.
Park, Joung-Jun;Yoo, Han-Jun;Choi, Hye-Won;Cheong, Ki-Soo;Kim, Ji-Tae;Park, Choon-Keun;Yang, Boh-Suk
한국수정란이식학회지
/
제27권4호
/
pp.253-258
/
2012
This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozen-thawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h ($94.2{\pm}2.6%$, $94.67{\pm}3.4$ and $129.67{\pm}5.5$). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to $98.3{\pm}1.7%$ after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.
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