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.

Internal sperm storage after mating is important for insect reproduction, because it permits delayed fertilisation, and post-copulatory mate choice in polyandrous females. The polyandry is common in many animal taxa including insects, because it increases female fitness by reducing the risk of infertility and providing opportunities for sperm competition and choice. The reproductive success of males, on the other hand, often depends upon avoidance of sperm competition by preventing mated females from copulating and receiving sperm from other male suitors. A widespread strategy used by males is the use of the male seminal fluid proteins (SFPs) that form the mating plug and alter female behaviors, for example by suppressing mating receptivity and elevating egg-laying. Under these circumstances, females are expected to evolve mechanism(s) to control exposure to the male SFPs in order to maximize fitness by balancing the positive and negative impacts of polyandry. Here, we discover that Drosophila melanogaster females eject male ejaculates 1-6 h after mating with a stereotypic behaviour, and that this is regulated by a brain neuropeptide pathway composed of diuretic hormone 44 (Dh44), and its receptor Dh44R1. We showed that suppressing Dh44 or Dh44R1 signals in the brain expedites sperm ejection, whereas enhancing Dh44 or Dh44R1 signals delays sperm ejection. This study uncovers a molecular mechanism by which females can influence sperm competition and selection, and counter actively the negative impact of polyandry.

The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted (30×106 spermatozoa/ml) in different semen extenders. Semen samples were stored at 17℃ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at 17℃.

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.

Preservation of sperm is essential for long-term storage of valuable animal genetic resources and for the conservation of threatened mammalian species undergoing progressive extinction. In this study, using pig as a model system, we evaluated the feasibility of germ-plasm preservation via sperm cell lyophilization. We show that, pig sperm can be successfully lyophilized and stored in a liquid nitrogen-free condition for at least 6 months. Intracytoplasmic injection of lyophilized sperm (ICSI), stored at 4℃ for four months, into in vitro matured pig oocytes could successfully develop up to blastocyst stage (13.0±3.0%). Lyophilized sperm could also be stored at room temperature for at least three weeks without further compromising their in vitro development up to the blastocyst stage (14.6±3.2 vs. 16.6±5.1%; p>0.05). Blastocysts produced from ICSI of lyophilized sperm stored at 4℃ or room temperature contained similar number of cells per blastocyst (44.9±3.2 vs. 44.0±4.3; p>0.05) but was significantly lower than those produced from non-lyophilized fresh sperm (52.1±5.8 p>0.05). Interestingly, use of a custom-designed HEPES-buffered, calcium-free, defined medium for the lyophilization resulted in normal post-ICSI embryonic development up to blastula stage (23.4±2.8 vs. 24.0±2.9%) and, the resultant blastocysts contained similar number of cells per blastocyst (47.9±4.3 vs. 50.6±7.0) compared to those generated from non-lyophilized fresh sperm (p>0.05). These lyophilized sperm could also be stored at room temperature for at least three weeks with slight reduction in post-ICSI embryonic development (19.6±1.4%). Therefore, these results suggest that, pig sperm could be successfully and efficiently lyophilized for their long-term storage at 4℃. Lyophilization of sperm could be a practical option for long-term storage of mammalian germ-plasm.

This study was undertaken to find out the effect of cholesterol and serum albumin on sperm ability and lipid peroxidation levels period to the liquid storage of miniature pig sperm. Ejaculated semen from miniature pigs was collected by gloved-hand method into a pre-warmed () thermos bottle, and extended with Modena solution {with and without BSA, methyl-beta-cyclodextrin (-cholesterol) and cholesterol loaded cyclodextrin (+cholesterol)}. Each semen was assessed for viability (SYBR-14/PI staining) and acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining at 1, 3, 5, 7 and 10 days of storage. At for the effects of cholesterol and serum albumin on lipid peroxidation, semen were incubated with (), and lipid peroxidation level were measured by flow cytometry using the lipid peroxidation reporter probe . The result, lipid peroxidation level in sperm added with cholesterol were lower in compared to the added sperm with serum albumin. Also, added cholesterol to sperm had significant (p<0.05) higher viability when storage for 7 and 10 days and lower when 10 days of storage percentage of acrosome-reacted sperm (AR pattern) in acrosome state as say result compared to other treated groups. In conclusion, role of cholesterol during lipid storage in miniature pig spermatozoa was protected boar spermatozoa from lipid peroxidation prior to lipid storage. Addition serum albumin during lipid storage in sperm may be induce sperm membrane damage by lipid peroxidation. Therefore, addition of cholesterol to miniature pig sperm will be lead to extension of liquid storage periods.

Antioxidants partially ameliorated the detrimental effects of reactive oxygen species (ROS) on sperm characteristics during in vitro storage. The objective of the present study was to investigate the single or synergetic antioxidative effect of curcumin and Vit. E on the characteristics of fresh boar sperm during in vitro storage. The sperm viability in curcumin, Vit. E supplementation and curcumin+Vit. E+H2O2 groups remained over 85.0% in 3 hr incubation period, but in 6 hr incubation period, curcumin+Vit. E+H2O2 groups was sharply dropped than those of curcumin and Vit. E group. The membrane intergrity in all evaluated groups except for H2O2 group did not significantly difference in 3 hr incubation period. The viability in curcumin or Vit. E supplementation were significantly increased than in curcumin+H2O2 and Vit. E+H2O2 group in 6 hr incubation period. The percentage of mitochondrial activity and acrosome intergrity obtained similar trends within same incubation periods irrespective of treatment. The lipid peroxidation of spermatozoal plasma membrane ranged from 11.6∼17.5 nM/l×106 and 14.0∼ 19.0 nM/l×106 in 3 hr and 6 hr incubation periods. In conclusion, curcumin or Vit. E rpplementation alone or cooperatively improved sperm viability index (motility, membrane intergrity, viability and survival rates) and fertility index (mitochondria activity, acrosome intergrity and lipid peroxidation) of fresh boar sperm, indicating that curcumin and Vit. E have a antioxidative properties through its scavenging activity against hydrogen peroxide.

In present study, attempts were made to preserve abalone (Haliotis discus hannai) sperm in liquid form at low temperature, to evaluate the effect of various diluents in short-term storage on sperm, and cryopreservation procedures were optimized for the cryoprotectants as well as freezing rates, in terms of the motility and survival rate, and the ultrastructural changes of sperm after short-term storage and cryopreservation were observed. The abalone sperm reached maximum motility until about 4min after activation. The motility was constant for about 16min, after which it dropped gradually, and about 50min later all motility ceased. Threshold activation of sperm was found in 40% artificial seawater (ASW), and motility increased as the concentration of ASW increased. In Hanks balanced salt solution without calcium (Ca-Free HBSS, 300 and 400 mOsmol/kg) and 10%, 20%, and 30% ASW the sperm was immotile, and motility once again restored incompletely only in HBSS of 300 and 400 mOsmol/kg, 20% and 30% ASW after 100% ASW was added. Sperm motility was extended following 20 days of cold storage only in 70% and 100% ASW. A high motility index of 3.5-4.5 was observed for the first 8 days in 70% and 80% ASW. In other diluents sperm motility was constant less than 10 days, and the motility index was obviously lower than that of sperm in 70% and 100% ASW. After 20 days of cold storage survival rates of 10.2%-20.7% were obtained in ASW and 300 mOsmol/kg HBSS, and that in 400 HBSS (65.3%) was significantly higher than others. The constant period of sperm motility stored in 70% ASW was longer obviously than that in 100% ASW after 6 days of storage, and the time to maximum motility of sperm stored in 70% increased gradually, while the difference in which of sperm in 100% ASW was not significant. The sperm plunged into liquid nitrogen all died except that sperm using 15% glycerol as cryoprotectant restored 10.4% of motility. The highest motility index (3.4) was obtained with 5% glycerol and freezing procedure: 50℃/min from to .

말똥성게 정자의 냉장보존을 위한 여러 가지 희석액의 보존 효과를 비교하였다. 다양한 농도의 포도당 용액, 인공해수(ASW)를 희석액으로 하여 정자를 4에서 보존하였다 보존효과는 정자 활성지수 (SAI), 생존율 및 말똥성 게 난에 대 한 수정률로 평가하였다. ASW와 1.2 M glucose액을 희석 액으로 하여 30일간 냉장보존하였을 때 가장 높은 운동성 및 생존율을 유지하였다. 정자의 냉장보존을 위한 희석액의 적정 pH는 7.0∼8.0 이었다. 정자