본 연구는 현재 국립축산과학원 가축유전자원세터에서 보유 하고 있는 재래닭을 순수화된 품종인 것으로 판단하고, 일반 적으로 이용되고 있는 산란사료 및 사양관리 방법을 적용하여 특히, 동절기에 있어 재래닭의 정자의 보존 기간과 수정률 및 초기배자의 생존율을 각각 비교함으로써 재래닭의 생산성 향 상을 위한 기초자료를 제공하고, 나아가 표준능력을 고찰하고 자 수행하였다. 본 시험에 사용된 공시계는 39주령의 재래닭 6계통 적갈색(R, Red Brown Strain), 황갈색(Y, Yellow Brown Strain), 회갈색(G, Gray Brown Strain), 흑색(L, Black Strain), 백색(W, White Strain) 그리고 오계(O, Ogol Strain)를 대상 으로 하고 대조군으로는 3계통 즉, 외래도입종 중에서 대표적 인 다산종인 White Leghorn (F Strain), Rhode Island (C Strain) 그리고 육용종인 Cornish (H Strain)의 수정률 및 초기배자 생존율을 조사하였다. 단 한번의 인공수정 후, 3 주간 생산된 알의 수정률 확인을 한 결과, 재래닭의 경우, 6 계통간의 유의 적인 차이는 없지만 93.3 ~ 100.0비율로 인공 수정 후, 2일째 부터 수정률이 6일째 동안 최고 높음을 확인했다. 6일째까지 상대적으로 일정하게 높은 수정률을 유지하다가 7일부터 17일 째 까지 점진적으로 감소함을 확인했다. 17일 이후 생산된 알 의 경우 무정란임을 확인 할 수 있었다. 재래닭(R, Y, 그리고 O) 21일간 생산된 알의 배발생정지율의 결과, 인공수정 후, 약 4일째 생산된 알(3 ~ 6일)에서 외래도입종 3품종간의 유의적인 차이는 보이지 않았지만, 배발생정지율이 0%임을 확인하였다. 세 품종 모두 약 7일째 생산된 알부터 배발생정지율이 13.8 ~ 26.7%로 급격히 증가하고 12일부터 감소하는 것을 확 인했다. 재래닭 3 계통의 결과도 인공수정 후, 약 4 일째 생 산된 알에서 외래도입종 3 품종과 유사한 패턴을 나타내면서 배아 사망율이 6 일째까지 0%를 보였다. 금후, 체내 정자보존 기간이 수정률 및 초기배자 생존율에 미치는 영향을 좀더 엄 밀하게 조사하기 위해서는 정액 성상 및 활력 검사와 더불어 암탉의 주령에 따른 변화도 함께 보다 세부적이고 입체적인 방법의 체계적인 조사가 반드시 필요하다고 할 수 있겠다.

Humulus japonicus is an ornamental plant in the Cannabaceae family. Although the mode of action of Humulus japonicus is not fully understood, a strong relationship was observed between anti-inflammatory and anticancer in some types of cells. Recent studies also have shown that Humulus japonicus possesses anti-inflammatory activities and may significantly improve antioxidant potential in Raw 264.7 macrophage cells. Thus, the aim of this study was eva-luated the effect of Humulus japonicus extract on sperm motility and subsequent preimplantation developmental com-petence of the bovine embryos. After in vitro maturation, the oocytes with sperms were exposed in in vitro fertilization (IVF) medium supplemented with Humulus japonicus extract (0.01, 0.05, 0.1 μg/mL, respectively) for 1 day. In our results, exposure of IVF medium to Humulus japonicus extract did not affect sperm motility and percentage of pene-trated oocytes but ROS intensity was significantly decreased by 0.01 μg/mL compared with other groups (p< 0.05). Moreover, treatment with 0.01 μg/mL of Humulus japonicus extract was higher the frequency of blastocyst formation than the any other groups (p<0.05). Otherwise, treatment with 0.01 μg/mL of Humulus japonicus extract not increased the total cell number but reduced apoptotic-positive nuclei number. In conclusion, our results indicate that supple-mentation of Humulus japonicus extract in IVF medium may have important implications for improving early embryo-nic development in bovine embryos

The main purpose of this study is to estimate the effect of adding Tea-N-Tris (TES) to the freezing buffer for miniature pig sperm. In particular, we attempted to identify the association between the MMPs expression and the fertility and viability of frozen sperm from each extender (LEY (Lactose Egg-Yolk), TLE (TES + LEY), TFGE (TES + Fructose + Glucose Egg-Yolk)). In accordance with this, Hypoosmotic Swelling Test (HOST) respond test was the lowest among sperms frozen in LEY while the highest HOST respond was observed among sperms frozen in TLE. Furthermore, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of MMP-9 and MMP-2 was the highest in sperms frozen in LEY, Meanwhile, sperms from the TFGE and TLE group showed lower level of MMP-9 and MMP-2 expression in the order of TLE being the lowest. LEY group showed lower rate of blastocyst development than the TES supplement group, although the difference was not statistically significant. Meanwhile the rate of blastocyst development appeared similar when sperms from TLE and TFGE group were used for IVF. Together, these results indicate that adding Tea-N-Tris to the sperm freezing buffer only suppresses MMPs protein activation but also maximize in-vitro fertility, providing a means to improve the success rate in the in vitro manipulation of miniature pig sperm.

본 연구는 돼지 정자의 동결 건조 시 동결 보호제인 trehalose의 효과와 동결 건조 시간과 동결 건조 후 저장 기간에 따라 정자의 생존성과 체외 성숙 난자 내 동결 건조 정자를 직접 주입한 후 전핵 형성율, 난할율 그리고 배발달 성적을 조사하였다. 동결 건조 후 정자의 생존율은 trehalose 무첨가구에 비해 trehalose를 첨가한 처리구에서 높은 생존율을 보였으며, 75 mM의 trehalose를 첨가하여 동결 건조한 정자들의 생존율이 가장 높은 것으로 나타났다. 또한, 동결 건조 후 저장 기간이 길어질수록 생존율이 낮아지는 경향이었다. 체외 성숙 난자 내 동결 건조 정자를 직접 주입 후 전핵 형성율은 trehalose 첨가구에서 유의적으로 높았으며(p<0.05), 난할율과 배발달 성적도 trehalose 첨가구에서 유의적으로 높았고(p<0.05), 정자의 동결 건조 시간이 짧을수록 높은 난할율과 배발달율을 보였다.

Cryopreservation of epididymal spermatozoa offers a potential tool for rescuing genetic material from males of genetically elite populations. Castration, catastrophic injury, sudden death or any other event that makes semen collection or mating impossible may prematurely terminate a stallion reproduction. Stallion epididymal spermatozoa vary widely in the loss of progressive motility, acrosomal integrity, and viability during freezing and thawing. The objective of this work was to investigate the effect of (1) freezing package types on cryopreservation efficiency, (2) thawing temperatures (37, 56 or 70℃) on Computer Assisted Sperm Analysis (CASA) parameters and (3) post-thawing incubation time (0, 1, 2 or 4h) on castrated stallion epididymis. Post-thawed sperm motility ranged between 59.69% and 64.28% (56℃ and 37℃) in various thawing temperatures. When stallion epididymis sperm was frozen, straw was better than freezing tube on VCL (Velocity of Curvilinear Line) and VAP (Velocity of Average Path) parameter. Higher percentage of motility was observed at 37℃ thawing temperature even though no significant difference was observed among various temperatures. The motility, VCL, ALH (Amplitude of Lateral Head displacement), VAP, BCF (Beat-Cross Frequency) and STR (Straightness index) parameter of post-thawed sperm were significantly decreased by increasing the incubation time for all thawing temperatures. The present study showed that type of freezing package (Straw vs. Freezing tube) was not significantly different on cryopreservation efficiency. Furthermore, stallion epididymal spermatozoa frozen-thawed at 37℃ for 1 min resulted the highest proportion of motility and velocity movement. In addition, motility and viability of frozen-thawed stallion epididymal spermatozoa were also decreased by incubation.

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG 0.2 μM, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% eggyolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide(H2O2) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under 30℃ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in 0.3℃. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group (43.3±4.7%) was significantly (p<0.05) higher than other treatment groups (Tissue: 16.3±2.7% and Water: 27.5± 3.1%), dying sperm (SYBR+/PI+) in Air treatment group (55.6±4.7%) was significantly lower than other treatment groups (Tissue: 77.6±3.2% and Water: 67.6±3.3%) (p<0.05). Acrosome reaction in Air treatment group (0.2±0.1%) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: 0.7±0.2% and Water: 0.5±0.1%), the acrosome reaction in Air treatment group (28.6±2.8%) within all sperm also was significantly lower than other treatment groups (Tissue: 44.2±1.8% and Water: 36.2±2.0%) (p<0.05). And mitochondrial intact in Air treatment group within live (97.1±0.4%) and all (61.9±3.3%) sperm were significantly higher than other treatment groups (Tissue: 85.2±3.3%, Water: 87.8±2.9% within live sperm and Tissue: 49.28±3.7%, Water: 42.0±3.1% within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol (39.85% ±11.41) than in 5% glycerol treatment (18.08%±1.61). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol (34.12%±11.02) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

Male factor infertility or sub-fertility contributed half of all cases of infertility while the semen abnormality is the current topic of argument. Conventional analysis of semen showed poor correlation with fertility. Therefore, evaluation of current semen analysis method is necessary to improve standards of semen assessment. The goal of this study was to investigate that correlation between motion kinematic before and after capacitation and litter size in porcine. Sperm motility and kinematics were measure by computer-assisted sperm analysis (CASA). The motility of spermatozoa was positively correlated with curvilinear velocity (VCL), average path velocity (VAP), and mean amplitude of head lateral displacement (ALH) (p<0.05). Where as VCL positively correlated with VSL, VAP and ALH (p<0.01). Straight-line velocity (VSL) was positively correlated with VAP and ALH (p<0.01). VAP was significantly positively correlated with ALH (p<0.01). Also, we found significant positive correlation among variation of VSL, VAP and ALH (p<0.05). No motility and kinematic parameter are correlated with litter size. However, litter size was significantly correlated with breed (p<0.05). Our results suggested that analysis of sperm motility and kinematics using CASA is questionable for prediction of litter size. However, it has some practical importance to evaluate semen commercially.

This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22～24 hr incubation from counting agar plate in which sperm dilute to 10 ～106 in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome- intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 (77.24±6.47, p<0.001) and 7 days (77.24±6.47, p<0.001) after preservation compared to 1 (15.71±7.18) and 3 days(18.39±7.22) after preservation, respectively. Sperm viability was significantly lower (53.25± 35.03, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 (15.71±7.18) and 3 (18.39±7.22) days compared to 5 (21.84±7.91) and 7 (22.59±9.93) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.

This study was designed to determine whether low-density lipoporoteins (LDL) extracted from egg yolk in extender improve the function of Korean Jeju Black Bull semen. The semen was cryopreserved with 5% ethylene glycol (EG) or 7% glycerol (G) extenders containing 10% egg yolk (EY), 4% LDL and 5% EY or 8% LDL. Frozen-thawed sperm were evaluated sperm motility, viability, membrane integrity and acrosome integrity. Post-thawed sperm motility has been significantly higher (p<0.05) in 4% LDL + 5% EY (; EG and ; 7% G) than 8% LDL (; EG and ;G). Treatment of 4% LDL + 5% EY-EG () has been significantly improved sperm viability compared to other treatments except 10% EY - EG. Moreover, in membrane integrity, swollen sperm ratio has been only significantly increased (p<0.05) in 4% LDL + 5% EY - EG () among all treatments. In assess to detect acrosome integrity, especially, AR pattern ratio has been significantly decreased (p<0.05) in 4% LDL + 5% EY - EG among all treatments. In sperm viability as time passes, between 4% LDL + 5% EY and 10% EY, there was no significant difference, but 8% LDL was significantly decreased sperm viability in EG (1 and 2 hrs) and G (30 min, 1, 2, 5 and 12 hrs) extender. However, there were no significant differences among all treatments except 8% LDL-G in sperm membrane integrity. 8% LDL-G has been significantly decreased swollen sperm ratio at 5 hrs after thawed. It is concluded from these results that 4% LDL + 5% EY to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Korean Jeju Black bull.

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

This study mainly inquired characteristics and changes of 'Chang-aelgool' through 38 cases(with 161 Ssang-chang) of annex and pavillion buildings in Yeong-nam region which are built during the Chosun dynasty. The method of inquiry included actual survey of windows along with bibliographical research, and the results are as below. First, through the discovery of the term 'Chang-aelgool' as an indication of the window-forming frame in 'YeongGeonUiGwe'(1680 A.D), it is apparent that the term 'Chang-aelgool' was widely used in Korea from the late 17th century. Second, the 'Chang-aelgool' of study objects are classified into 4 categories. Type Ⅰ and Ⅱ are comprised of mitre-joints which cover the 4 corners of 'Chang-aelgool' and mainly used in building annex and pavillion buildings during the early period of the Chosun dynasty. Type Ⅲ was widely used during the early and middle period of the Chosun dynasty and drastically dropped in number during the late period of the dynasty. Type Ⅳ is comprised of mitre-joint of the upper-half, tenon-jointing of the lower-half and widely used in annex and pavillion building during the late period of the Chosun dynasty. Third, the form of 'Chang-aelgool' has changed from rectangular form with longer width during the early period of Chosun dynasty to square form during the middle period and eventually ended up as a rectangular form with longer height during the late period of the dynasty. Fourth, it is considered that while mullion which is located in the center of 'Chang-aelgool' was mainly used around the main floored room during the early period of the Chosun dynasty, became commonly used in main floored room and 'ondol' rooms during the middle period and drastically dropped in number from then and ended up being not in use after the mid 18th century.

닭 정액의 주요성분인 glutamine은 정장의 주요 성분으로 정자의 생리 활성에 매우 중요 한 것으로 알려져 있으며 조류 정액 희석액의 주요성분으로 이용되어져 왔다. 그러나 glutamine은 액상의 배양액에서 변성이 일어날 수 있어 세포배양에는 alanyl glutamine을 이용하 여 장기 세포배양에 이용되고 있다. 본 연구에서는, alanyl glutamine을 이용하여 개발된 희 석제를 이용하고 닭 정액의 냉장보존에 이용하였으며, 닭 정자의 장기 생존성에 미치는 영 향을 조사하였다. 국립축산과학원 가축유전자원시험장에서 사양관리중인 오골계 수탉 또 는 레그혼에서 복부 마사지법으로 채취된 정액을 이용하였다. 채취한 정액은 96 mM L- Glutamine 또는 Alanyl Glutamine을 함유하는 희석액에 각각 1:8의 부피비율로 상온에서 희석하 여 상업용 냉장고와 4℃로 조절된 BOD 배양기에 보존을 실시하였다. 냉장되는 온도의 저 하 속도를 낮추기 위하여 상온에서 희석하는 온도와 동일한 온도의 증류수 300 ml을 비이 커에 담고 물위에 1.5 ml E-tube에 담아서 파리필름으로 밀봉하였다. 희석된 정자의 활력은 상업용 냉장고에 보존할 경우, 4일차까지 75.6%의 활력정자가 유지 되었으나 5일 이후에는 급격히 생존성이 저하되었으며 6일째에는 활력 정자의 수가 18%로 낮게 나타났다. 한편, BOD 배양기에서 보존된 동일한 정자는 9일까지 약 73%의 활력정자가 유지되었으며, 10일 이후에는 활력 정자의 수가 급격히 감소되는 현상을 관찰하였으며, 15일째에는 약 20%의 정자 활력이 유지되는 현상을 관찰하였다. 이러한 정자의 활성 유지는 정자의 미토콘드리 아 에 필요한 글루타민 에너지 공급원을 안정적으로 공급하는 기능에 의하여 유지 되는것으로 추정되며 특히 hyper activated sperm의 비율을 줄여주어 정자군의 노화를 방지할 수 있는 효과가 있는 것으로 추정되었다. 그러므로 조류 정액을 냉장 보존할 때, 알라닐글루타민을 희석액의 주요 성분으로 이용될 수 있음을 보여주고 있다.

칡한우는 한우의 품종의 하나로 FAO에 등록되어 있고, 1,700여두가 사육되고 있는 것으 로 추정하고 있다. 그러나 사육두수가 적어 멸종위험품종으로 분류되고 있고, 특히 번식 가 능한 수소의 숫자가 매우 적어 근친의 위험에 노출되어 있다. 칡소에 대한 연구는 유전적 특성(이 등, 2002), 정액동결(조 등, 2011), 모색분포(박 등, 2007) 등으로 아주 제한적으로 보고되고 있어서 증식에 필요한 정액 동결관련 연구가 필요할 것으로 생각된다. 따라서 본 연구에서는 칡한우 수소의 활용도를 높이기 위하여 생후 15개월령 단계에서 2-3일 간격으 로 2회 채취하여 동결한 후 거세·비육하여 도체성적을 평가하였다. 정액채취는 인공질(FHK, Japan), 동결희석제는 Triladyl(Minitube, Germany) 희석제를 이 용하였다. 정자의 motility는 CASA system(Minitube, Germany)상에서 Total motile (TM)과 Progressive motile (PM) 비율을 측정하였다. 15개월령 칡소와 30개월령 칡소의 신선 및 동결융해 정액의 활력을 조사한 결과, 15개월 령 수소의 정액량이 평균 2.3 ml이었고, 30개월 수소의 정액량은 평균 5.0 ml로서 유의하게 높았다(p<0.05). 채정 직후의 신선정자의 TM 비율이 15개월 및 30개월 수소가 각각 평균 93.7% 및 88.3%, PM 비율은 각각 97.0% 및 88.3%로서 PM 비율은 15개월령이 유의하게 높았다(p<0.05). 한편 동결융해 정자의 TM 비율은 각각 평균 56.0% 및 58.0%였고, PM 비 율은 각각 평균 64.0% 및 70.7%로서 차이가 없었다. 칡소 15개월 정액채취가 완료된 후 거세 및 비육하여 출하한 결과와 한우 거세우의 도축 성적을 비교한 결과, 육량은 A등급이 칡한우는 35.7%, 한우는 27.5%였고, B등급은 각각 42.9% 및 52.75였다. 육질등급은 1++등급이 각각 35.7% 및 15.0%, 1+등급이 각각 35.7% 및 30.8%, 1등급은 각각 14.3% 및 32.0% 그리고 2등급이 각각 14.3% 및 20.1%였다. 본 연구를 통하여 칡한우 수소 생후 15개월령에서 정액채취, 동결 및 활용이 가능한 것 으로 판단되며, 향후 칡한우 고유 유전자원의 확보 및 증식에 활용도가 높을 것으로 기대된 다.

닭 정액의 동결보존기술은 유전자원 보존의 수단으로 이용될 수 있는 기술로서 고병원성 조류인플루엔자 같은 악성질병에 의한 멸실을 방지하는 매우 중요한 기술이다. 닭의 정자 는 동결보호제와 동결에 이용되는 희석액에 따른 융해 후 정자의 활성도, 수정율 및 부화율의 변이가 매우 큰 것으로 알려져 있다. 가장 널리 사용되는 동결보호제인 Glycerol은 동결 정 액 제조에 가장 많이 이용되며 융해 후 닭 정자의 생존성이 비교적 우수한 것으로 알려져 있으나, 인공수정에 직접 이용할 경우에 수정율의 저하현상이 나타난다. 본 연구에서는 동 결보호제중 하나인 N-Methylacetamide(MA)를 이용하여 한국 재래계인 오계 수탉의 정자 를 동결보존, 융해 및 인공수정을 실시하여 수정율과 부화율을 조사하였다. 복부 마사지법으 로 채취한 정액을 5℃ 저온수조에 담아 실험실로 이송하였으며 MA을 함유하지 않은 희석액 (HS-1)에 1:1의 비율로 희석하여 30분간 5℃ 온도로 유지시켰다. 2차 희석액은 14, 18 그 리고 22%의 MA가 함유된 희석제를 이용하였으며 1차 희석된 정액과 1:1 비율로 희석하여 0.5ml 스트로에 충진 하였다. 액체질소 표면 위 4 cm에 설치된 간이 지지대에 밀봉된 스트 로를 30분간 정치하여 동결하는 간이동결법을 이용하였다. 동결된 정액을 1～2주간 보관한 후, 5℃로 조절된 저온수조에서 융해하고 운동성 있는 정자와 활력이 우수한 정자의 비율 을 비교 분석하였다. 동일한 방법으로 융해한 정액으로 인공수정을 실시하고 7일 간 수정 란을 수집하여 부화기에 배양하여 7일차에 발생란을 검란하였다. 대조군에서 희석된 정액 의 경우 98.4% 수정율이 관찰되었으며 실험군에서는 7, 9 및 11% MA의 경우 각각 21.5 %, 34.7% 및 25% 수정율이 관찰되었다. 수정율에 대한 부화율은 대조군에서 88.9%를 관 찰하였고 실험군은 100%, 89.5% 및 87.5%로 관찰되었다. 이러한 결과로 유추해 볼 때, 오 계 정자의 동결보존에 유용할 수 있는 MA의 농도의 범위는 약 9% 내외로 판단되며 특히 MA는 오계 수정란의 수정율과 부화율에 미치는 영향이 낮은 것으로 사료되었다.