본 연구는 엘크 암사슴(Cervus canadensis)의 사육방식에 따른 사료섭취량, 체중변화 및 자록의 성장에 미치는 영향을 확인하고 적정 방목강도를 구명하기 위하여 수행되었다. 본 실험에 사용 된 공시가축은 3~4년생 엘크 암사슴 16두(평균체중 : 236.2 ± 15.7 kg)를 이용하였으며, 방목초지는 3년 이상된 톨 페스큐 위주의 기성 혼파초지로서 초종구성은 톨 페스큐(약 50%), 오차드 그라스(약 10%), 켄터기 블루그라스(약 5%)와 피, 바랭이 등으로 구성되었다. 방목초지의 수분함량은 19.51∼22.61%로 방목기간 동안 유사하였으며, 조단백질 함량은 6~7월에 유의적으로 높게 나타났다. 조지방과 조회분 함량은 가을로 갈수록 점차 증가하였으며, NDF와 ADF함량은 각각 53.65∼60.18%과 26.08∼29.10% 로 조사되었다. 실험기간동안 보충사료 섭취량은 월별로 처리구 사이에 유의적인 차이가 나타나지 않았으나, 조사료와 총 건물섭 취량은 5월을 제외하고 처리구 사이에 유의적인 차이가 나타났다 (p<0.05). 특히 8월에는 GR처리구의 조사료 섭취량이 급격히 감소하였는데, 여름철 환경변화에 따른 것으로 사료된다. 반면, 9월 포유를 위해 부족한 영양소를 섭취하기 위해 사초섭취량이 급격히 증가하였다. 우리 연구에서 암사슴의 채식패턴은 수사슴과 차이를 보였으며, 이는 실험기간 자록의 이유 및 포유기간에 따른 영향을 받은 것으로 사료된다. 본 실험에서 사양방식에 따른 엘크 암사슴의 체중은 유의적 차이가 나타나지 않았으나, GR처리구에서 자록의 성장률이 빠른 것으로 나타났다. 그러나, GR처리구에서 BF처리구에 비해 낮은 이유율이 나타나 분만 전후 사사사육을 통해 이유율을 높여주는 것이 필요할 것으로 판단되며, 목지 내에서 분만 시 포유경험이 있는 산차가 높은 암사슴을 이용하고 방목강도를 높여주는 것이 이유율을 향상시키는 데 도움을 줄 수 있을 것으로 사료된다. 엘 크 암사슴의 방목강도는 연평균 15두/ha로 나타났는데, 이는 다마 암사슴(Dama dama)에 비해서는 체중대비 높게 나타났으나, 엘크 수사슴에 비해서는 낮은 수치를 보여주었다. 이러한 결과는 기성초지 이용으로 인해 사초의 생산성이 감소한 결과로 사료되며, 앞으로 방목초지의 혼파방법과 기성초지의 비배관리 등을 통해 초지 활용성을 높일 수 있는 연구가 추가적으로 필요할 것으로 사료된다.

In this study, the effect of forage sources in the total mixed ration (TMR) on in vitro goat rumen fermentation was investigated. Rice straw (RS), Italian ryegrass (IRG), timothy (TIM), and alfalfa (ALF) were used as forage sources. Each forage source was mixed with a commercial goat concentrate diet in the ratio of 1:1. Total 4 TMR were prepared. Rumen simulated in vitro fermentation using goat rumen fluid collected from the slaughterhouse was conducted until 72 th . For fermentation parameters, gas production (GP), volatile fatty acids (VFAs), and ammonia nitrogen (NH3-N) were examined. All assays were performed at 24 th , 48 th , and 72 th h of incubation individually. Contents of crude protein and non-fibrous carbohydrate were greater in the order of RS < IRG < TIM < ALF. Significant treatment effects were found in valerate and NH3-N at 24 th h of incubation (p<0.05). ALF showed the greatest contents of them and RS was the lowest. At 48 th incubation, a significant effect was detected at GP (p<0.05) and RS was greater than others. However, GP of RS was lower than others at 72 th . Significant effects on Total VFA, butyrate, and valerate productions were found at 72 th h of incubation (p<0.05). ALF showed the greatest production. Methane production from all treatments was not significantly different for each incubation time (p>0.05). The present study provided primary information on how goat rumen fermentation responds to different nutrient contents and forage sources of TMR. And the information could be used for the design or optimizing economical diet formulation for goats.

엘크 수사슴의 체중 성장곡선 및 일당증체량을 추정하기 위하여 국립축산과학원 가축유전자원센터에서 사육중인 엘크 수사슴 183두의 체중-일령 자료 1,509개에 대해 비선형 회귀식인 Gompertz와 Logistic 모형을 적용하였다. 추정된 엘크 수사슴 성장곡선 모수인 성숙체중(A), 성장비율(b)과 성숙률(ｋ)은 Gompertz 모형의 경우 각각 335.5±1.415kg, 2.18±0.045와 0.0025±0.00005였고, Logistic 모형에서는 330.5±1.337kg, 5.46±0.209와 0.0355±0.00009였다. 적합도 검정을 위한 결정계수(R2)는 두 모형에서 큰 차이가 없어 두 모형 다 적합한 것으로 나타났다. 다만, 실제 측정값과 추정값에 대한 차이인 평균제곱근 오차 값이 엘크 암사슴에 비해 크게 나타났으며 따라서 엘크 수사슴의 낙각 및 번식기 체중 증감을 고려한 성장곡선 모형에 대한 추가적인 연구가 필요할 것으로 판단된다. 엘크 수사슴에서 Gompertz 모형으로 추정된 함수식은 f(t)=335.5Xe^-2.18e^-0.0025t였고, Logistic 모형의 경우 F(t)=330.5X(1+5.465e^-0.00355t)^-1였다. 각 모형에서 제1차 도함수로 유래된 일당증체량 곡선은 Gompertz 모형의 경우 F'(t)=0.0025Xf(t)X1n(335.5/f(t)), Logistic 모형의 경우f'(t)=0.0036Xf(t)X[1-f(t)/330.5]였다. 이 함수식을 통해 도출된 엘크 수사슴에서 최대 증체량을 나타내는 시기인 315~478.3일령, 123.4~165.3㎏ 도달 전후에는 보다 세심한 사양관리가 필요할 것으로 보인다. 본 연구에서 도출된 엘크 수사슴의 성장곡선 모수 추정값, 변곡점 및 변곡점 도달일령 등은 사슴산업 종사자들이 사양관리 및 육종계획을 수립하는 데에 도움이 될 수 있을 것으로 사료된다.

The objective of this study was to estimate the growth curve parameters of body weight for female Elk. Weight and age data from 115 does raised at the Animal Genetic Resources Research Center in Korea were used in this study. The growth curve parameters were estimated from a nonlinear regression using Gompertz and Logistic models. Mature weight (A), growth ratio (b) and maturing rate (k) of female Elk were 214.1±2.17 kg, 2.12±0.0045 and 0.0043±0.00011, respectively, according to the Gompertz model. They were 208.3±2.17 kg, 5.56±0.234 and 0.0065±0.00017, respectively, according to the Logistic model. The goodness of fit determined by R2 was higher in the Gompertz model than that in the Logistic model. The growth curve functions obtained from the Gompertz and Logistic models in female Elk were f(t)=214.1×e-2.124×e-0.00484×t and f(t)=208.3×(1+5.561×e-0.00651×t)-1, respectively. The absolute growth rate functions from the Gompertz and Logistic models in female Elk were f'(t)=0.0043×f(t)×1n(214.1/f(t)) and f'(t)=0.0065×f(t)×[1-f(t)/208.3]respectively. The growth pattern of female Elk generated from this study can be useful in determining the most appropriate feeding plans and the best breeding strategies for deer.

Cryopreservation of germ cells from genetically proven animals could be a source of restoration tools from the risk of extinction or disappearance of wanted characteristics. Using frozen semen, the genetic gains of Korean native cattle have been increased greatly for 70 years. The preservation of genetic resources as a form of frozen semen straw has limited availability due to the numbers. To circumvent this weakness of frozen semen, we tested two re-freezing methods with different initial thawing temperatures using frozen Korean proven semen and rare breed semen from albino, black and chikso breeders. It has been known that human sperm could resist to cryo-damages by repeated freeze-thaw cycles, but not for Korean proven bulls number (KPN) or for rare breeds. Total 7 frozen semem from brindled(2), black(1), Korean Albino(2) and KPN(1) bulls were used for our research. After thawing straws under 5°C/2min or 37°C/40sec with low temperature water bath and thermo jug, spermatozoa were re-diluted with triladyl diluents after first thawing and re-frozen. Sperm motilities were compared between animals and treated groups after re-thawing. Mean values of motility and viability of refrozen/thawed sperm for expansion of the number of straws were significantly higher in 5°C than in 37°C (P< < 0.05). However, the activity of viable sperm thawed at 5°C was significantly decreased after first and second thawing. It is estimated that re-freezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa.

난자생검(Ovum pick up, 이하 OPU) 기법은 유효축군 조기확보 및 우수한우 유전자 원의 조기증식, 희소유전자원 다량확보를 위해 매우 중요한 기법으로 주목받고 있다. 이 러한 OPU기술에는 체외배양기술 외에도 현장의 환경과 시기에 따라 회수되는 난자의 수 역시 중요하다. 현재, 가축센터에서는 한우의 유전자 다양성 확보를 위한 일반 한우 외에 도 희소한우 증식을 위한 연구를 진행 중에 있다. 본 연구는 OPU기법을 활용하여 가축 유전자원센터에서 보유중인 한우 또는 칡소를 대상으로 계절별 및 모색별로 난포란의 개 수와 난자회수율의 차이를 비교 · 검토하고자 실험을 진행하였다. 실험시기는 3월에서 5 월(봄), 6월에서 8월(여름), 9월부터 11월(가을)까지 구분하여 실시하였으며, 계절별 및 모 색별로 채취되는 난자의 개수와 회수율을 분석을 하였다. 봄에는 8.20±6.57, 여름에는 5.47±4.01, 가을에는 5.88±3.65개가 회수되었으며, 또한 계통별로는 가을에 실시한 결과 한 우는 6.40±6.15개, 칡소는 2.75±0.55개가 회수되었다. 그리고, 계통별로 수정란발달률은 한 우는 난할률이 25.0±44.4%, 배반포발달률이 6.3±50.0%, 칡소의 경우 난할률이 27.3±21.0%, 배반포발달률이 9.1±16.7%로 나타났다. 계절별로 OPU를 실시한 결과 여름, 가을경에 실 시하는 것에 비해서 봄에 실시를 하였을 경우 회수되는 난자의 수가 많은 것으로 확인을 하였다. 그리고, 현재 일반한우가 칡소에 비해서 회수되는 난자가 더 많은 것으로 확인되 었다. 이에 반하여 난할률 및 배반포발달률은 차이가 없는 것으로 확인되었으나, 향후 실 험진행에 따라 정확한 결과를 도출할 것으로 사료된다.

본 연구는 칡소의 동결정액 생산 시 항산화제 첨가가 정자 생존성 및 운동성 등에 미 치는 영향을 조사하고자 수행하였다. 공시축의 사양관리는 한우사양관리(국립축산과학원, 2007) 기준에 준하여 사육하였다. 공시축은 정액채취를 위해 승가훈련이 완료된 칡소 5두 를 공시하였고 정액채취 빈도는 주 1회로 하였다. 정액 채취는 암소 보정 후 공시축의 승가를 유도한 다음 인공질에 음경을 삽입하여 채 정 후 원정액 정자가 80∼90% 생존율을 나타내는 것을 동결정액 생산에 활용하였다. 동 결보존 희석액의 기본조성은 Tris(hydroxymethyl)amino-methane 250mM, Citric-acid monohydrate 80mM, Fructose 60mM, Penicillin G Streptomycin 1%, Egg-Yolk 20%, Glycerol 7%에 항산화제인 Ascorbic-acid를 무처리, 0.5mM, 1.0mM, 2.0mM로 농도로 하 여 본 실험에 공시하였다. 동결정액의 생산은 처리군별로 원정액을 1차 희석한 다음 저온 실로 이동하여 2차 희석 및 Glycerol 평형을 유도한 다음 0.5ml Straw에 충진시켜 액체 질소를 활용한 동결을 실시하였다. 융해 후 정자 특성분석은 위상차 현미경과 컴퓨터정자 분석기(Computer Assisted Sperm Analysis)를 활용하여 생존율, 운동성, 선형성, 직진도 등을 검사한 결과 생존율은 2.0mM, 무처리, 1.0mM, 0.5mM(83.3%, 81.6%, 65%, 60%)였 고 운동성은 무처리, 2.0mM, 1.0mM, 0.5mM(97%, 96.5%, 94.4%, 91.3%)순이였고, 선형성 은 2.0mM, 무처리, 1.0mM, 0.5mM(29%, 29.7%, 32.9%, 36.4%)순으로 낮은 경향을 보였 다. 직진도는 2.0mM, 무처리, 1.0mM, 0.5mM(65.6%, 62.5%, 62.2%, 60.4%)순으로 낮은 경향을 보였다. 동결보존 희석액에 항산화제인 Ascorbic-acid 첨가함으로써 정자의 대사 과정에 따라 생성되는 활성산소의 발생 억제 등으로 인하여 동결융해 후 생존율에 영향 을 미치는 것을 감안하여 분석한다면 항산화제의 농도별로 각각 상이한 결과를 나타낸 것은 Ascorbic-acid의 첨가가 정자의 동결 융해 후 생존율 및 운동성 등에 영향을 미쳤 을 것으로 판단된다. 이와 같은 결과를 바탕으로 칡소 동결정액 생산 시 생존율 극대화를 위해 적정 항산화제 및 첨가 농도를 확립하고 이를 활용한다면 가축유전자원 보존·증식 에 기여할 것으로 사료된다.

Historically, Korea old cattle had been consisted with various lines of coat color brindle, black and white-brown breeds or more. The two rare lines of black and white coat color are maintained for animal resources and preserved critically. The present study was carried out to evaluate potential usage of cysteamine supplementation during in vitro matration (IVM) and in vitro culture/production of embryo (IVP) by transvaginal ultrasound-guided follicle aspiration (Ovum Pick-Up: OPU) for the establishment of cryo-banking system. Immature slaughterhouse-derived cumulus-oocyte complexes (SL-COCs) were matured in IVM medium supplemented with 0, 0.1, 0.3 or 0.9 mM cysteamine, and then cultured in mSOF-BAS for 8 days after in vitro fertilization. The treatment of 0.1 mM cysteamine on SL-COCs showed higher rate of blastocyst, so OPU-derived COCs from rare breeds were matured in TCM media supplemented with or without 0.1 mM cysteamine, FSH and 5% FBS. The embryos were evaluated their developmental stages on day 8. During IVM, cysteamine treatment significantly increased the embryo production rate of slaughterhouse-derived COCs (19.6% vs. 30.5%). The presence of cysteamine during IVM of OPU-derived COCs from rare Korean cattle breeds (albino white and black line) also increased embryo production rates than those from SL-COCs (27.4% vs. 41.9% and 36.4%). With these results, cysteamine treatment during IVM is one of key factors IVP of blastocysts to establish banking system of endangered rare Koarean cattle with OPU derived transferable blastocysts.

During the ovary preservation in low temperature, the cumulus oocyte complexes(COCs) lose their developmental competences after in vitro fertilization. We used phosphate-buffered saline (PBS) as a basic solutions of at various temperatures (25, 15 or 5 ℃) and supplemented them with 1mM glucose and 0.5mM glutamine as a source of carbohydrate metabolites. After recovery of COCs and in vitro fertilization, a significantly higher number of oocytes developed into blastocysts. The developmental competence of embryos that were originated from ovaries preserved at 15 ℃ was increased compared to those of 25 or 5 ℃. The maturation rate of oocytes was not differed between 24 and 36 h at 15 ℃ but showed lower than control group (71% versus 78%). In vitro-fertilized oocytes from ovaries stored at 25 ℃ for 24 h or at 5 ℃ for 24 h had a significantly decreased developmental potentials, but at 15 ℃ did not (27% versus 29% of blastocysts to develop into day 8). With these results, bovine ovaries can be preserved at 15 ℃ for 36 h without decreasing developmental capacity of in vitro-fertilized oocyte at least to the blastocyst stage. This information provides valuable information of preserving ovaries for embryo transfer or in vitro embryo production.

The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be 6.20±2.28/donor from the control group and was significantly lowly recovered to be 1.57±1.72/donor from the group treated at a sperm concentration of 10×106 (p<0.05). The number of unfertilized embryo was 0.8±1.30/donor in control group which was significantly lower than the group treated at a sperm concentration of 4×106 (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

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±15.1), 1,2-PROH (88.0±21.1) or the control (99.9±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±1.3), EG (12.1±1.1), Glycerol (15.2±1.8), and 1,2-PROH (11.5±1.3) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.5±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.

The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was 20×106/straw. Sexed X frozen semen with 2×106 cells or 4×106 cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was 24.9 ± 7.31% and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between 2×106 cells or 4×106 cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with 2×106 cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

It is generally accepted that chronic stress impairs female reproduction. It negatively affects ovarian function and the number of ovulated oocytes. Chronic stress lowers the number of retrieved oocytes. Ovarian follicular development is regulated by both pituitary-derived gonadotropins and intraovarian regulatory factors. The main corticosteroids are cortisol, cortisone, 11-deoxycortisol and corticosterone, cortisol being one of the most commonly used welfare and stress physiological indicator. In this study, we investigated the effect of cortisol level on progesterone patterns and ovulation in the dog. Cortisol and progesterone level of serum were analyzed by radioimmunoassay. The day of ovulation was considered as the day when serum progesterone concentration was 6.0∼8.0 ng/ml. In vivo dog oocytes were collected by flushing oviducts of mixed-breed bitches at three days after ovulation. We classified dogs as having group 1 (cortisol level, 0 ≤ or < 2 μg/dl), group 2 (corisol level, 2 ≤ or < 4 μg/dl), group 3 (cortisol level, 4 ≤ or < 6 μg/dl) and group 4 (cortisol level, 6 μg/dL ≤). The patterns of progesterone were not different in four cortisol groups. The average numbers of retrieved oocytes was not different in four cortisol groups. These results suggest that different cortisol levels on estrus dogs do not affect ovulation, number of ovulated oocytes and progesterone changes.

The objective of this study was to monitor health conditions of four genetically identical somatic cells cloned Labrador retriever puppies by estimation of body weight and analysis of hematologic and serologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and four live puppies were delivered by Caesarean section. The cloned Labrador retrievers were genetically identical to the nuclear donor dog. The body weight of clone-1, -2, -3, and -4 was increased from 0.66, 0.40, 0.39, and 0.37 kg at birth to 6.2, 6.6, 6.2, and 6.0 kg at 8 weeks of age, respectively. Although clone-4 had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned puppies were not different when compared to reference range. In serological analysis, Glucose, ALP and inorganic phosphate level of four cloned puppies was significantly higher than the reference ranges. However, there was no significant difference among four cloned dogs. This study suggests that cloned puppies derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological and serological parameters.