• 한국수정란이식학회지
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• 제31권4호
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• pp.349-353
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• 2016

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.

• 한국수정란이식학회지
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• 제9권1호
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• pp.23-29
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• 1994

The long term goal of this research is to develop an efficient procedure for large scale production of genetically identical or cloned animals. To improve nuclear transpalntation efficiency in the rabbit, this study evaluated the age of nuclear recipient oocytes on the different steps of nuclear transplantation. The ovulated oocytes in different ages were collected from the superovulated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) supplemented with 10% fetal calf serum(FCS) from 13 to 15, 17 to 20 and 23 to 26 hours after hCG injection. The denuded oocytes were used as nuclear recipient cytoplasm following enucleation by micromanipulation. The blastomeres separated from the 8-cell embryos were used as nuclear donor. The enucleated oocytes receiving a blastomere in the perivitteline space were fused in the 0.28 M mannitol solution at 1.5 kV/cm, 60 sec for three times. The fused oocytes were co-cultured with the monolayered rabbit oviductal epithelial cells in TGM-199 solution with 10% FCS for 72 hours at 37$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos and in vivo developed embryos collected at 72 hours after hCG injection were stained with Hoechst 33342 dye. Their cell numbers were counted under a fluorescent microscope. The results obtained were summarized as follows ; 1. The aged oocytes(20 hrs. post hCG) showed significantly(P<0.05) higher fusionrates(70 ~ 90%) than the recently ovulated oocytes(30.8%) 2. The aged oocytes which were electrically activated and fused at 20 hours developed to blastocyst at significantly(P<0.05) high rate, while none of the recently ovulated oocytes developed to blastocyst. 3. Even though the aged oocytes at 23~26 hours showed higher fusion rate(85.7%), not only they were inadequate to manipulate but also their developmental potential to blastocyst was highly impaired. 4. The developmental potential in vitro of nuclear transplant embryos was significantly retarded than in vivo deveolped embryos.

• 한국수정란이식학회지
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• 제30권3호
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• pp.213-218
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• 2015

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

• 한국수정란이식학회지
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• 제29권3호
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• pp.305-311
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• 2014

It is imperative to know the reproductive parameters of the existing swine breed for profitable farming in Bangladesh. This study was designed to determine the reproductive parameters of sows at hilly areas. A total of 245 pigs with 47 breedable sows were included. The data on age at puberty, oestrous cycle length, oestrus duration, gestation length, interval between furrowing and onset of oestrus, first service pregnancy rate, service per pregnancy and number of piglets born per sow in local and cross bred sows were determined. The present study revealed that age at puberty of local and cross breed sows was $232.5{\pm}8.4$, $221.3{\pm}6.9$ days, respectively. Oestrous duration was $41.1{\pm}3.1$ hours. The interval between furrowing and onset of oestrus was shorter in cross bred sows. There was no variation in values of oestrus cycle length and gestation length. The first service pregnancy rate was higher in both the local and cross bred sow. Number of piglets per sow per farrowing was $6.1{\pm}2.2$ and 60.9% local sows gave birth of 3 to 5 piglets per farrowing whereas 75.0% cross bred sows gave birth of 6 to 9 piglets. These results suggest that reproductive parameters of local sow need to be improved for better production, and cross bred sows should be reared for obtaining expected productivity.

• 한국수정란이식학회지
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• 제28권4호
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• pp.325-335
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• 2013

The study was set for one year to measure the effects of concentrate supplementation on reproductive performances and semen quality in indigenous rams. The study was conducted at the Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh during the period from May 2011 to April 2012. Forteen ram lambs (4~5 months) were randomly divided into two equal groups (n=7); supplemented vs control. The animals of control group were maintained on natural grazing. Along with natural grazing the supplemented group was on supplemented feeding. The concentrate supplementation (Wheat bran, Crushed maize, Soy bean meal, Fish meal, DCP powder, Vitamin mineral premix, Salt) was provided @ 300 g/head /day to the supplemented group. Body weight, scrotal circumference, BCS and libido index were measured weekly. Age, body weight and scrotal circumference at puberty were recorded. Semen was collected once in a weak using artificial vagina and chilled at $5^{\circ}C$ for 48h for evaluation. Concentrate supplementation did not influence (p>0.05) body condition score, age, weight, scrotal circumference at puberty and libido index. Final body weight (kg), growth rate (g/d), scrotal circumference (cm) and scrotal growth rate (mm/15d) were significantly (p<0.05) higher in supplemented group of rams compared to control. Volume, concentration, motility and membrane potentiality of spermatozoa were varied significantly (p<0.05) in supplemented and control groups. However, density, mass motility, viability and sperm with normal acrosome, midpiece and tail were not differed insignificantly (p>0.05) in different observation times. It was concluded that concentrate supplementation with free grazing improved weight and scrotal circumference gain and semen production with increased quality in indigenous ram.

• Clinical and Experimental Reproductive Medicine
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• 제44권2호
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• pp.85-89
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• 2017

Objective: The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. Methods: We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. Results: There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p<0.05). The implantation/pregnancy rate (IVF, 50.93%; ICSI, 55.10%), miscarriage rate (IVF, 12.57%; ICSI, 16.29%), and live birth rate (IVF, 42.12%; ICSI, 44.08%) were similar (p>0.05). No cycles were canceled due to the formation of no usable blastocysts. Conclusion: Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.

• 한국초지조사료학회지
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• 제9권2호
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• pp.62-67
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• 1989

A breeding program in progress at Suweon Livestock Experiment Station, RDA, involves the hybridization of italian ryegrass (Lm) and several leading forage grasses in an effort to combine the nutritive, productivity and palatability qualities of Lm with the adaptive and cold resistance qual; .les of several leading grasses. In order to study the fate of the hybridization between remotely related species, immatured hybrid embryos were cultured on media. The emasculated plants of Lm were Sikem and Tetrone. Reweille, 2n=14, of perennial ryegrass (Lp.), Forager, 2n=42, of tall fescue(Fa), First, 2n=14,of meadow fescue(Fp), Potomac, 2n=28, of orchardgrass(Dg), and Richmond, 2n=42, of timothy(Ph.p) were used as pollinators. Embryos were isolated on 4, 8, 12, 16, and 20 days after pollination and cultured them on modified Murashige and Skoog media. Calluses and plantlets have been obtained after 8 days old embryos crossing between $Lm{\times}Fa$, after 12 days embryos crossing between $Lm{\times}Lp$, Dg, and Ph.p, and after 16 days embryos crossing between $Lm{\times}Fp$. Both callus and shoot formation occurred on 6 % or less of the plated embryos of $Lm{\times}Fp$, Dg, and Ph.p. Embryoderived callus forming shoots have been obtained from 4 days old embryo crossing between $Lm{\times}Dg$ which has not successfully been done anywhere as far as we know. It means that hybrid plants of species without crossability can be obtained through the use of immatured embryo culture. Some of plated embryos developed directly shoots from embryos and the others shoot-forming callus. Cross between related species showed a high frequency of directly shoot formation from plated embryos and cross between remotely related species a high frequency of callus formation.

• 한국수정란이식학회지
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• 제12권2호
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• pp.171-179
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• 1997

The objectives of the present study were improvements in the efficiency of developmental rates to morula and blastocyst stages to produce a large number of genetically identical nuclear transplant embryos. The oocytes collected from slaughterhouse ovaries were matured for 24 h and then enucleated and cultured to allow cytoplasmic maturation and gain activation competence. And then the donor embryos were treated for 12 h with 10 $\pi$g /ml nocodazole and 7.5 $\pi$g /ml cytochalasin B to synchronize the cell cycle stage at 26 h after the onset of culture. The blastomeres were transferred into the perivitelline space of the enucleated nocytes and blastomeres and oocytes were fused by electrofusion. The cloned embryos were then cultured in various conditions to allow further development. The age of the recipient(30 vs 40 h) had no significant effect on the fusion rates(82.4 vs 82.1%) and the developmental rates to morula /blastocyst(9.8 vs 11.0%). Effect of Nocodazole treatment on the donor cell cyle synchronization to improve the developmental rates of bovine nuclear transplant embryos was significantly higher than control group(21.4 vs 10.1%, p<0.05). Significant differences were in the percentage of fusion rates(72.9,77.1vs 61.9%) in three types of fusion medium(PBS(+), mannitol and sucrose, p<0.01). The developmental rates of bovine nuclear transplant embryos appeared to be highest in mSOF medium under 5% 0$_2$ condition, but no significant differences were found when compared with TCM199-BOEC and mSOF under two different oxygen ratio(5 and 20%).

• Clinical and Experimental Reproductive Medicine
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• 제28권4호
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• pp.265-270
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• 2001

Objective : The aim of this study was to evaluate the outcome the GnRH antagonist (Cetrotide) short protocol in controlled ovarian hyperstimulation comparing with GnRH agonist long protocol. Materials and Method: From July 2000 to November 2001, 26 patients, 28 cycles were performed in controlled ovarian hyperstimulation by GnRH antagonist and GnRH agonist. GnRH antagonist (Cetrotide) was administered in 12 patients (14 cycles, Group 1) and GnRH agonist (Lucrin, Sub Q, Group 2) in 14 patients (14 cycles). Ovulation induction was performed by hMG (Pergonal) in group 1, and by Combo (Metrodine HP + Pergonal) in group 2. We compared the fertilization rate, good quality embryo, and clinical pregnancy rate between the two groups. Student-t test and Chi-square were used to determine statistical significance. Statistical significance was defined as p<0.05. Results: Ovarian hyperstimulation syndrome did not occurred in which estradiol (E2) level was $3874{\pm}809\;pg/ml$ and the number of retrieved oocytes was $18.4{\pm}2.4$. The number of used gonadotropin ampules was significantly decreased in Group 1 (26.0 vs. 33.1, p<0.04). There were no significant difference in the number of preovulatory oocyte ($10.6{\pm}6.9$ vs. $10.0{\pm}6.1$), fertilization rate ($74.8{\pm}23.4$ vs. $72.2{\pm}21.8$), good quality embryo ($58.7{\pm}23.6$ vs. $38.7{\pm}36.6$), and embryo transfer ($4.3{\pm}1.6$ vs. $4.4{\pm}1.6$). Although the age of the group 1 was older than the group 2 (34.4 vs. 30.8), there was no significant difference in clinical pregnancy rate (50.0% vs. 57.1%). Conclusions: We suggest that GnRH antagonist was a safe, effective, and alternative method in the controlled ovarian hyperstimulation, especially in PCOD patients who will be develop the ovarian hyperstimulation syndrome.

• 한국수정란이식학회지
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• 제13권3호
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• pp.219-226
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• 1998

본 연구는 토끼난자에 있어서 배란 후 경과시간에 따른 탈핵률을 조사하고 ionomycin과 DMAP을 사용하여 활성화와 아울러 자체 탈핵을 유도하는 방법을 고안하였으며 아울러 이들의 탈핵효율과 핵이식후 체외발달을 확인한 결과는 다음과 같다. 1. hCG 주사후 15∼16 시간에 채란된 토끼난자는 73.4%의 탈핵률을 보였고, 16∼17시간에는 75.8%, 19∼20 시간에는 58.5%의 탈핵률을 보여 토끼난자의 탈핵은 hCG 주사후 17시간 이내에 실시함이 효과적임을 알 수 있었다. 2. 제1극체와 극체주위 염색질을 제거하므로서 핵을 제거하는 기존방법에서는 70.4%의 탈핵률을 보였지만 개선된 방법을 사용할 때 96.8%의 탈핵률 보여 탈핵률이 항상되었다. 3. 개선된 탈핵방법으로 핵이식하였을 때 88.2%의 핵융합률과 33.3%의 배반포 발달률을 보여 기존의 탈핵방법과 대등한 효과가 인정되었으며 활용성이 있음을 확인하였다.