• Journal of Embryo Transfer
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• v.21 no.3
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• pp.225-231
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• 2006

The objective of this study was to determine changes in serum hormone concentrations, blood chemical values and recovery rate of in vivo embryos during the estrous cycle following super-ovulation treatments in Jeju black cows. Superovulation was induced by subcutaneous administration of FSH twice a day for 4 days. Serum hormones were assayed by radioimmunoassay (RIA) and blood chemical values were analyzed by blood analyser system. Embryos were collected from all treated black cows using nonsurgical technique on day 7 after artificial insemination (AI). The results of this study were summarized as follows: 1. The progesterone concentrations were $7.2{\pm}3.8ng/ml$ at day -11 and $0.3{\pm}0.1ng/mL$ at day 0 (Day 0 is the first day of AI). The estradiol concentrations were $10.6{\pm}4.48pg/ml$ at day -11 and $15.0{\pm}2.2pg/ml$ at day 0. The lowest level of progesterone was measured at day 0. The highest levels of estradiol was measured at day 0. 2. The blood chemical values of treated black cows were no significant differences in normal cow values. 3. Sixty two embryos were collected in 12 black cows. Among the collected embryos, 37 embryos (59.7%) could be transferred into recipients. These results would be used as the basic informations for changing patterns of hormonal level and blood biochemistry in Jeju black cow with superovulation.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.263-267
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• 2013

Pig producers have been shown keen interest of the number of spermatozoa in a semen dose since pig artificial insemination introduce. However, determining the minimal number of spermatozoa need per AI without detrimental effect on overall reproductive performances is not an easy question to answer. To increase the efficiency of semen utilization in pig AI, optimum number of spermatozoa per dose needed to determine. The objective of this study was to determine the reproductive performance and factors that affect on-farm application of low-dose semen insemination in sows. Data were collected from Darby Genetics AI studs from 4th of June to 7th of July, 2012 (n=401). The numbers of parturition were 84, 234 and 83 in sows inseminated with doses of $1.5{\times}10^9$, $2.0{\times}10^9$ and $2.5{\times}10^9$ spermatozoa in 100ml extender, respectively. There were no significant differences on reproductive performances such as gestation period, total born, total born alive, stillbirth and mummy in sows inseminated with different semen doses. The average number of born alive was 10.5, 11.0 and 10.4 from sows inseminated with $1.5{\times}10^9$, $2.0{\times}10^9$ and $2.5{\times}10^9$ sperms, respectively. Also, number of spermatozoa per dose did not affect litter size (p>0.10). There were no significant differences of maternal genetic line difference on gestation period, total number born, number born alive, born dead and mummy. The estimated correlation coefficients of the different semen doses with total number born, number born alive, born dead and mummy were r=-0.00, -0.01, 0.02 and 0.02, respectively. Taken together, the result of this study suggested that when semen was appropriately inseminated after induced ovulation, insemination with low-dose ($1.5{\sim}2.0{\times}10^9$) semen dose not adversely affect sow's fertility.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.375-383
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• 2000

The nuclear maturation and developmental competence of immature, oocytes collected from donors at various morphology of corpus luteum (CL) and fertilized in vitro was investigated by comparing the meiotic activity and the yields of embryos. Ovaries were divided and classified into 4 groups as the following criteria : Group 1 ; ovaries showed evidence of recent ovulation (corpus hemorragicum). Group 2 ; apex of CL was red or brown. Vasculization was limited to periphery of CL. Group 3 ; apex of CL was orange or tan. Vasculization was covered over apex of CL. Group 4 ; CL was light yellow to white and firm in texture and the vascular network on the surface of CL had disappeared. Modified TCM 199 was used for maturation in vitro of immature oocytes and development was induced by using TLP-PVA as a basic medium. When oocytes collected from each group of donors had been matured for 4, 14, and 24 hours in vitro, the proportion of oocytes reaching metaphase I and metaphase II were not different among oocytes from 4 group of ovaries. Mature metaphase II stage of oocytes in each group was first observed at 14 hours, whereas completion of maturation of. oocytes in each group was at 24 hours. Luteal morphology of ovaries had little effect on the proportion of embryos reached 2 cells and 8 cell stage. However, the proportion of embryos cleaved to morula and blastocyst stage was significantly higher in the oocytes obtained from group 1 and 3 than in the oocytes from group 2 and 4 (p＜0.05). This data suggest that reproductive status of the donor significantly influence the yield of in vitro embryos.

• Journal of Aquaculture
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• v.5 no.2
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• pp.117-126
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• 1992

Induced breeding and indoor rearing of the catfish, Silurus asotus were performed. Ovulation was induced by injection of 5 mg dried carp pituitary per kg body weight. Fertilization rates were $81.5\~98.0{\%}$ and hatching success was $67.0\~82.0{\%}$. There were no significant differences of daily growth rate (DGR) and food conversion ratio (FCR) between the groups fed extruded eel pellet and extruded carp pellet. No significant differences in DGR and FCR were also found between $3{\%}$ and $4{\%}$ daily feeding rates. DGR and FCR of small fry were slightly affected by dissolved oxygen (DO) level in aquaria. When the DO level maintained at $4.5\;mg/{\ell}$, the best result was obtained and DGR and FCR were $5.7{\%}$ and 0.65, respectively. At the higher level of DO, $5.5\;mg/{\ell}$, DGR and FCR were $5.52{\%}$ and 0.77, respectively, whereas at the lower level of DO, $3.5\;mg/{\ell}$, the DGR was decreased to $3.45{\%}$ and FCR was increased up to 1.01.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.925-936
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• 2005

The present study evaluated whether pentoxifylline added to the freezing extender could improve the sperm characteristics and function in canine frozen semen. Also the conception rate following AI with frozen-thawed semen was investigated. The beneficial effects of pentoxifylline supplementation were visible in motility, viability, acrosome reaction, and tail swelling patterns. Especially, highest sperm viability and function were obtained in the forzen semen supplemented with 1mM pentoxifylline. The follicle size measured by ultrasonography was 6.48 mm, 11.52 mm and 8.9 mm on 11, 13 and 15 days after the onset of natural estrus, respectively and ovulation occurred on 13 and 15 days. The pregnancy rates in bitches inseminated with frozen semen on natural and induced estrus were 71.4% and 75.0%, respectively. There was no significant difference between the pregnancy rates in bitches inseminated with frozen semen following natural and induced estrus, but the litter size was slightly increased in natural cycle.

• Journal of Embryo Transfer
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• v.24 no.1
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• pp.33-37
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• 2009

Multiple ovulation and embryo transfer (MOET) has the potential to increase the rates of genetic improvement in cattle. Thus this study was performed to investigate several factors influencing in vivo embryo production in Holstein cattle under field conditions. The donors were superovulated with Folltropin-V and $PGF_2{\alpha}$ combination method. From Day 10 onward, donors were superovulated by i.m., twice daily, administration of 400mg Folltropin-V given in a series of decreasing doses over a 4-day period: on the first day, 3.5ml; on the second day, 3.0ml; on the third day, 2.0ml; and on the fourth day, 1.5ml (20ml in total, equivalent to 400mg of NIH-FSH-P1). Estrus was induced by i.m. administration of 25mg prostaglandin $F_2{\alpha}$ on the sixth and seventh of FSH treatment. Estrus detection was performed twice daily beginning 24h after the first prostaglandin $F_2{\alpha}$ injection. Donor cows were artificially inseminated 12 and 24 h after first standing estrus with semen from a proven Holstein sire. Embryos used in this study were recovered Day 7.5 of the cycle (Day 0: first standing estrus). From 195 superovulated dairy cows, 2,104 eggs were recovered, of which 1,172 were classified as transferable embryos based on morphological evaluation of quality. The results are summarized as follows: 1. The numbers of recovered and transferable embryos did not significantly differ among the capacity of milk production that were < 10,000kg/305days (group 1), $10,000{\sim}12,000\;kg$/305days (group 2) or > 12,000kg/305 days (group 3) (p>0.05, Table 1). 2. No differences in the numbers of recovered and transferable embryos were found among the donor's postparient days (p>0.05, Table 2). 3. Also, the numbers of recovered and transferable embryos of each superovulation seasons did not significantly differ among the four groups (p>0.05, Table 3).

• Clinical and Experimental Reproductive Medicine
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• v.15 no.2
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• pp.103-117
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• 1988

Ovulation induction was done with 3 different regimens as clomid combined with HMG, HMG only, and FSH combined with HMG in 28 menstrual cycles for IVF-ET and GIFT program. The appearance of endogenous LH surge, estradiol plateau, atypical LH surge, and time from initiation to peak of LH surge in serum and urine were observed and compared in 3 groups. 1. The estradiol concentration of serum LH surge day was similar in three groups but 1st group (Clomiphene Citrate+Sequential HMG) was slightly higher at $1924.0{\pm}865.1\;pg/ml$. In regards to the existence of serum estradiol plateau, 3rd group (FSH+Sequential HMG) was highest at 60%, and 1st group and 2nd group (HMG only) were similar at 33% and 44% respectively. 2. The number of ovarian of ovarian follicle which was more than 18mm in diameter was $4.1{\pm}2.0$, $4.2{\pm}2.1$ respectibely for 2nd group and 3rd group. Although the numbers were slightly higher thean 1st group for each ovarian follicle, serum estradiol value per follicle was higher for 1st group at $583.0{\pm}261.2pg/ml$. 3. When measuring the urine LH surge according to Hi-Gonavi and according to the standard set by three different types of surge, simultameous satisfaction for 1st group, 2nd group, 3rd group was two cases, five cases, four cases respectively at 40%, and the remained cases were composed of numorous type combination which satisfied the two definition, simultaneously in this study, the LH surge starting time was determined only in the cases tow combination were satisfied simultaneously at first, but there are something to study more. In one case of the 3rd group. 4. The concentration of LH surge start in urine and serum of 2nd group was highest at306. $0{\pm}287.2IU/l$ and $34.0{\pm}9.9mIU/ml$ and 1st group was low at $116.6{\pm}66.1IU/l$ and 7.4mIU/ml. The urine and serum value of LH was highest at $1644.4{\pm}988.8IU/l$, $65.9{\pm}15.0mIU/ml$ for 2nd group, 1st group was low at urine, and 3rd group was low of serum. With pregnancy established, the LH concentration of urine was relatively high but on the contrary the LH concentration of serum was low compared to urine concentration. 5. Time from LH surge start to the maximun of urine and serum value was highest at 15. $7{\pm}9.1$ hrs and $10.8{\pm}4.9$ hrs for 1st group and 3rd group. With pregnancy established, time was shortened for urine but on the contrary serum showed an increase in time. 6. The concentration of LH which increases with time on urine was highest at 2nd group ($194.6{\pm}76.8\;IU/hour$). The lowest increase for serum was at 3rd group (2.1mIU/hour). With pregnancy established, urine showed more increase than control group ($266.5{\pm}47.4\;IU/hour$) and for serum there was similar increase ($3.4{\pm}0.8\;mIU/hour$). 7. There were two examples of non-typical surge from 1st group and 3rd group each. Among these three cases showed a reestablishment of LH surge after 10-24 hours. 8. Endogenous spontaneous Lh surge occurs once for each 2nd group and 3rd group.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.195-200
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• 2015

The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female $F-{_1}$ mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG ($81.1{\pm}15.1$), 1,2-PROH ($88.0{\pm}21.1$) or the control ($99.9{\pm}21.3$) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO ($14.7{\pm}1.3$), EG ($12.1{\pm}1.1$), Glycerol ($15.2{\pm}1.8$), and 1,2-PROH ($11.5{\pm}1.3$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.5{\pm}0.7$, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

• Korean Journal of Agricultural Science
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• v.13 no.1
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• pp.82-89
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• 1986

This experiment was carried out to determine the optimum freezing and thawing rates of the hamster embryos. The female hamsters were induced to superovulate by intraperitoneal injections of 30 i.u. PMSG and mated with males of the same strain of 4 days the PMSG injection. They were killed and embryos were flushed from the oviduct and uterine horn on 3 days after mating. Embryos were flushed with a modified Dulbecco's phosphate-buffered saline and equilibrated with 1.5 M-dimethylsulphoxide by a 3-step procedure. The freezing rates of the samples were $1^{\circ}C/min$ from room temperature to $-6^{\circ}C$ and the samples were seeded at $-6^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were $0.3^{\circ}C/min$ from $-6^{\circ}C$ to $-35^{\circ}C$. From $-35^{\circ}C$ to $-70^{\circ}C$, the rates were divided into $0.1^{\circ}C/min$, $1^{\circ}C/min$ and $10^{\circ}C/min$, respectively. At $-70^{\circ}C$ the samples were plunged directly into liquid nitrogen. The samples were thawed at $4^{\circ}C/min$ and $12^{\circ}C/min$ from $-196^{\circ}C$ to $37^{\circ}C$, and for 2 min in $37^{\circ}C$ water bath, respectively. The average numbers of ovulation points and embryos recovered were 35.1 and 27.0 appearing 77.0% recovery rates. Eight cell embryos in the embryos recovered were 24.8. The survival rates of embryos according to the freezing rates were 55.5~67.7% at $0.1^{\circ}C/min$, 58.8~64.9% at $1^{\circ}C/min$ and 40.5~44.7% at $10^{\circ}C/min$, respectively. The survival rates at $10^{\circ}C/min$ were significantly low. The survival rates of embryos according to the thawing rates were 53.5% at $4^{\circ}C/min$, 53.7% at $12^{\circ}C/min$ and 59.1% in $37^{\circ}C$ water bath. The survival rates, in $37^{\circ}C$ water bath were slightly higher, but we did not find any differences among them. In conclusion, the best freezing rates of hamster embryos were $1^{\circ}C/min$ from the room temperature to $-6^{\circ}C/min$, $0.3^{\circ}C/min$ from $-6^{\circ}C/min$ to $-35^{\circ}C$ and $-0.1^{\circ}C/min$ or $1^{\circ}C/min$ from $-35^{\circ}C$ to $-70^{\circ}C$. The hamster embryos thawed for 2 min in $37^{\circ}C$ water bath showed the best survival rates.