• Journal of Animal Science and Technology
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• 제62권4호
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• pp.521-532
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• 2020

The production performance of broiler breeder hens in response to different levels of total lysine during the early laying period was investigated. A total of 126 Ross 308 parent stock hens were offered one of seven dietary treatments formulating elevated contents of total lysine ranging from 0.55% to 0.79% (0.04 scale; 133 g of feed) from 23 to 29 weeks of age. Each treatment had six replicates with three birds per pen. Body weight was recorded triweekly and eggs were collected and weighted at 9:00 am daily. One hen from each pen was euthanized to collect blood samples and visceral organs were harvested and weighed. Egg production, egg weight and egg mass were lower (p < 0.05) in hens offered a diet containing 0.55% total lysine compared to those fed the diet containing higher total lysine. Hens offered a diet containing 0.71%, 0.75%, and 0.79% total lysine had greater (p = 0.008) egg production rate compared to those offered a diet containing lysine less than 0.71%. The number of total eggs produced tended to be greater (p = 0.083) in hens offered a diet containing 0.71 and 0.75% total lysine compared to the other treatments. The number of settable egg production was higher (p < 0.001) in hens offered a diet contacting 0.79% total lysine compared to those fed the diet containing lower levels of total lysine. The relative weights of oviduct and ovary were lower (p < 0.05) in hens offered a diet containing 0.59% total lysine compared to the other treatments. No difference found in body weight, the number of total eggs, double-yolk eggs and abnormal shell eggs among the treatments. The urea nitrogen, estradiol-17 beta and progesterone in plasma were not affected by treatments. Based on linear- and quadratic-plateau models, total lysine requirements for egg production, settable egg production and egg mass at the early laying period were to be 0.73%, 0.77%, and 0.71%, respectively. Modern broiler breeder hens likely require higher total lysine than NRC recommendation in a diet for enhancing productivity during the early-laying period.

• 한국수정란이식학회지
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• 제25권3호
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• pp.141-144
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• 2010

In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

• 한국수정란이식학회지
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• 제23권4호
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• pp.269-274
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• 2008

The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.

• 한국수정란이식학회지
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• 제23권4호
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• pp.263-267
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• 2008

The purpose of this study was to determine toxic effect of sucrose and trehalose prior to cryopreservation on nuclear maturation and embryonic development in immature bovine oocytes. All cryoprotectant was prepared in tissue culture medium 199-HEPES (TCM 199-HEPES) with 10% fetal bovine serum (FBS). Immature oocytes were exposed to 1.2M ethylene glycol (EG) and 0.1M sucrose or 1.2M EG and 0.1M trehalose for 3 min and then were exposed to 3.2 M EG and 0.25 M sucrose or 3.2 M EG and 0.25 M trehalose for 1 min. Oocytes treated with cryoprotectants were exposed to 0.25 M sucrose or 0.25 M trehalose for 5 min and then 0.1 M sucrose or 0.1 M trehalose for 5 min. Depending on type of sugar added to cryopreservation solution, oocytes were allocated to sucrose group and trehalose group, respectively. Oocytes exposed to TCM 199-HEPES with 10% FBS were considered as control. Oocytes were cultured in TCM 199 supplemented with 10% FBS, 5 ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone, and $1\;{\mu}g/ml$ estradiol for 24 h in $39^{\circ}C$, 5% $CO_2$. Nuclear maturation was assessed by staining oocytes with 1% aceto-orcein. Oocytes were fertilized in vitro and were cultured in TCM 199 supplemented with 10% FBS, 5 mM sodium pyruvate, and antibiotics in $39^{\circ}C$, 5% $CO_2$. The rates of cleavage and blastocyst, and cell number in blastocyst were assessed. Metaphase II rates were not different among experimental groups regardless of type of sugar. The cleavage rate of trehalose group (73.3%) was significantly higher (p<0.05) than those of sucrose group (62.8%) and control group (60.8%). The blastocyst rate was significantly higher in trehalose group (p<0.05). Mean cell number in blastocyst were not different among experimental groups, although cell number of blastocyst in trehalose group was significantly higher on day 7 (p<0.05). In conclusion, sucrose and trehalose were not toxic to immature bovine oocytes prior to cryopreservation. In particular, trehalose was more effective on embryonic development.

• 대한수의학회지
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• 제29권3호
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• pp.373-381
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• 1989

The aim of the present study was to test the efficiency of estrous induction in the premature, metestrous and anestrous bitches. The estrus was induced with prostaglandin $F_{2{\alpha}}$, estradiol-$17{\beta}$, pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin(HCG) in the treatment A, and with PMSG and HCG in the treatment B. Day 0 was the first day of estrone injection in the treatment A and the day of PMSG injection in the treatment B. Twenty three of the twenty six bitches were laparotomized under general anesthesia between 11 and 18 days after onset of behavioral estrus, whereas three bitches were not laparotomized and remained until parturition. Ovarian responses were evaluated with the total number of corpora lutea or ovulation sites. The uterine horns were flushed with phosphate-buffered saline added heat treated canine serum(10%), the flushing media was collected into watch glass and the ova were examined under stereomicroscope. The results obtained were as follows: 1. Standing estrus was observed on the day $17.7{\pm}1.5$ after injection of estrone in the treament A, but ovarian responses were not detectable. 2. Standing estrus was observed on the day $12.2{\pm}0.2$ after injection of PMSG in the treament Band 14 of 17 bitches showed ovarian responses. Ova were recovered in 9 of the 14 bitches. 3. Ovarian responses were observed in one of the three premature bitches. two of the three metestrous bitches and all of the 11 anestrous bitches. The average number of the ova collected from 9 bitchs were $12.2{\pm}1.4$. 4. Three bitches in the treament B exhibited behavioural estrus and all of them were mated with fertile male dog, resulting the pregnancy in only one bitch. The pregnant bitch gave the birth of two pups.

• 한국수정란이식학회지
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• 제22권2호
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• pp.101-105
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• 2007

본 연구는 한우와 홀스타인 젖소에서 연속적인 다배란 처리 시 효율성 및 부작용에 대한 문제점을 연구하고자 다른 종간 다배란 처리효과를 비교, 조사하였다. 연속적인 다배란 처리를 위하여 estradiol benzoate(EB)와 CIDR plus, FSH 호르몬을 이용하여 한우에서는 17두, 젖소에서는 9두를 2회 이상 반복 이용하였다. 다난포 발생을 위해서는 FSH 호르몬을 이용하고 호르몬 처리 동기화를 위해서는 CER plus 기구를 질 내 삽입하였으며, 주요 조사 항목으로는 번식기관의 문제점, 배란 효율, 회수된 수정란 수, 우수 수정란 수 등을 분류하고, 종합적으로 종간 연속적인 다배란 효과를 비교, 조사하였다. 한우에서는 17두 중 4두(23.5%)에서 자궁경관의 협착이 발생하였고, 자궁 내에 손상이 발생하여 채란이 되지 않는 경우도 4두(23.5%)의 발생이 조사되었으나, 상대적으로 젖소에서는 9두 중 4두에서 일시적으로 배란이 전혀 일어나지 않는 경우가 있었다. 연속 처리 후 채란된 수정란의 수를 종합해 보면, 젖소에서는 평균 $4.3{\pm}3.4$개, 한우에서는 $2.7{\pm}2.5$개였으며, 종간 호르몬 효율과 연관된 배란율 조사에서는 한우에서는 89.7%, 젖소에서는 80.4%의 성적을 보였다. 또한, 3회까지 연속 다배란 처리를 한 결과, 한우나 젖소에서 채란된 수정란의 수는 비슷한 수준으로 채란되었다. 이와 같은 성적을 종합해 볼 때 EB제나 CIDR plus, FSH를 이용하여 1개월 주기로 연속적인 수정란 채란을 위한 다배란 처리가 소의 종에 무관하게 가능하다고 사료되지만 일시적으로 배란이 일어나지 않는 소에 대한 문제나 번식기관의 손상, 채란되는 수정란의 수가 적다는 것에 더 많은 연구가 필요하다고 사료된다.

• 한국동물학회지
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• 제39권3호
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• pp.257-265
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• 1996

참개구리를 사용하여 난소내 여포들이 성장시기에 획즉하게 되는 스테로이드들의 생성능력을 조사하였다. 개구리로부터 중간(0.7-1.11mm) 크기 이하의 여포들을 분리한 다음 전구스테로이들이나 뇌하수체추출물(FPH)을 포함한 배양액에서 6시간 배양 후 이들에 의해 산물스테로이드로 전환된 양을 방사면역 측정법으로 조사하였다. FPH의 처리는 중간 크기의 여포만 스테로이드들의 생성을 현저히 촉진하고 작은 여포들(0.35-0.7mm)에서는 촉진효과가 매우 미약하였다. Progenenolone의 첨가는 가장 작은 여포 (0.22-0.4mm)들에 의해서도 progesterone(P$_4$)으로 전환되었다. 그러나 가장 작은 여포에서는 P$_4$를 첨가했을 때에도 역시 같은 현상이 나타났다. 작은 여포와 중간 여포들에 의한 전환된 스테로이드들의 양을 단위표면적의 전환량으로 계산 비교해 본 결과 전환된 양은 거의 같았다. 이는 작은 여포의 여포세포들 (granulosa cells)이 이미 E$_2$에 이르는 모든 중간 산물들을 생성할 능력을 가졌다는 것을 의미한다. 그러나 이들이 FPH에 의해서는 스테로이드를 조금밖에 생성하지 못하는 것으로 보아 아직 부족한 또다른 요인들이 있는것으로 추정된다.

• Toxicological Research
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• 제27권3호
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• pp.181-184
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• 2011

Screening of estrogenic activity on dichloro diphenyl trichloroethane (DDT), dichloro diphenyl dichloro ethylene (DDE), dieldrin, heptachlor, aldrin, chlordane, lindane, polybrominated diphenyl ethers (PBDE) and parabens was compared using Organization for Economic Cooperation and Development (OECD) test guideline 455 (TG455). The estrogenic activity of DDT was 58,000-fold ($PC_{50}$, $1.67{\times}10^{-6}$ M) less than $17{\beta}$-estradiol($E_2$) ($PC_{50}$, $2.88{\times}10^{-11}$ M) but DDE, dieldrin, heptachlor, aldrin, chlordane, lindane and PBDE did not show any estrogenic activity in this assay system. In the case of paraben compounds, the rank of relative transcriptional activation (logRTA) was butyl paraben -1.63752 ($PC_{50}$, $1.25{\times}10^{-7}$ M) > isobutyl paraben -2.34008 ($PC_{50}$, $6.3{\times}10^{-7}$ M) > ethyl paraben -2.64016 ($PC_{50}$, $1.26{\times}10^{-6}$ M) > isopropyl paraben -2.73993 ($PC_{50}$, $1.58{\times}10^{-6}$ M) > propyl paraben -2.84164 ($PC_{50}$, $2.0{\times}10^{-6}$ M). Our data suggest that OECD test guideline TG455 may be useful as a screening tool for potential endocrine disruptors.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.31-31
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• 2003

The incidence of reproductive abnormalities in the male has been reported to have increased during the past 50 years. These changes may be attributable to the presence of chemical with oestrogenic activity in our environment. Present study was carried out to determine the effects of maternal exposure to xenoestrogens on the testicular development and on the transcriptional expression of the steroidogenic enzyme and subunits of inhibin/activin in testis of male offspring. Pregnant female mice were administrated with 4-tert-octylphenol (OP; 2, 20, 200mg/kg), Bisphenol A (BPA; 2, 20, 200$\mu\textrm{g}$/kg), $\beta$-estradiol 17-valerate (EV; 2$\mu\textrm{g}$/kg) or vehicle (CV; corn oil) during gestational days 11 to 17. Offsprings were sacrificed on gestational day 18 (fetal 18) and neonatal day 7. Body weights were significantly increased in groups treated with all doses of OP and BPA. Maximum seminiferous tubules diameter on gestational day 18 were not changed in any treatment group, however, they were significantly increased on the neonatal day 7 in the group treated with low-dose of OP (2 mg/kg) and BPA (2 $\mu\textrm{g}$/kg). Increased expression of the P450$_{17a}$-hydroxylase dehydrogenase (P450$_{17a}$), 3$\beta$-hydroxylase dehydrogenase (3$\beta$-HSD), and 17$\beta$-hydroxylase dehydrogenase (17$\beta$-HSD) on gestational day 18 were observed in the groups treated with 2 or 20 mg/kg of OP. However, expression of the steroidogenic enzymes were not changed in the groups treated with all the doses of BPA. In contrast with the results from fetal testis, no expressional changes of these enzymes was found in all the OP-treated group and increased expression of inhibin/activin $\beta$B subunit mRNA were obseued in the 200 $\mu\textrm{g}$/kg BPA-treated group in the neonatal day 7. These results suggest that gestational exposure to low level of xenoestrogen causes a stimulatory effects on the transcriptional expressions of steroidogenic enzymes and subunits of inhibin/activin and on the seminiferous tubule development by their estrogen-like actions.ons.

• 한국수정란이식학회지
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• 제10권1호
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• pp.45-51
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• 1995

In order to determine the optimum condition and timing for in vitro maturation of oocytes to metaphase of meiosis II (M II), the immatured follicular oocytes were recovered by puncturing the large(1.0~1.5 mm in diameter) and small(<1.0 mm in diameter) follicles in the ovaries of rabbits treated intramuscularly with a single dose of 100 TU PMSG 68 hours previously. The follicular oocytes were classified into three grades by the attachment of cumulus cells. The Grade I and II follicular oocytes from large follicles were cultured in BO-DM medium with 10% FCS, 35 $\mu$g /nl of FSH, 10 $\mu$g /ml of LH and 1 $\mu$g /ml of estradiol-17$\beta$ at 39t in a 5% $CO_2$ incubator for 11 to 23 hours. In 3 hours interval during the culture period, the oocytes were harvested and their cumulus cells were removed with hyaluronidase. The denuded oocytes were stained with Hoechst 33342 dye and their meiotic status and extrusion of the first polar body (PB) were examined under a fluorescence microscope. Also the fragmentation of the first PB and the distance between the first PB and nucleus were examined. The results obtained were as follows: 1. The mean recovery rate of follicular oocytes from the large and small follicles was 59. 9 and 31.3%, respectively. The mean number of oocytes recovered per rabbit and the Grade I percentage were 14.6 and 94.4% in large follicles, but 2.1 and 61.1% in small follicles, respectively. All the parameters examined were different significantly (p<0.05) between both the folliclular size. 2. Most of the follicular oocytes(86.8%) were matured in vitro to M II phase in 14 hours in Grade I oocytes, but the significantly(p<0.05) less oocytes(45.5%) were matured in Grade II oocytes. 3. The first PB was extruded in most of the oocytes(94.7%) in 14 hours of culture with the fragmentation rate of 29.6%, but the fragmentation rate of the first PB increased significantly (p<0.05) as the culture period for maturation was longer to 20 hours(63.5%). 4. The distance between the first PB and nucleus was increased linearly (p<0.05) as the maturation time passed from 14(7.1$\mu$rn) to 23 hours(58.4$\mu$m). 5. From the above results it was concluded that the optimum time for in vitro maturation culture might be 14 hours in the follicular oocytes from rabbit primed with PMSG for 68 hours, expecially when these follicular oocytes were used for recipient cytoplasms in embryo cloning.