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.

For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at —7 ℃ to —35 ℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

The objective of this study was to investigate the motility and kinematics of boar sperm that while stored at 4C. The samples of fresh boar semen were place into an extender, Androhep, and stored at . In three of these samples, cryoprotectants were added. The sperm's motilities and kinematics were evaluated by using microscope () and the viability status was evaluated by using with eosin staining method. The 5 sample groups are; Goup A:Androhep (extender), stored at . Group B:Androhep (extender), stored at . Group C:Androhep (extender), + 3% glycerol (cryoprotectant), stored at . Group D:Androhep (extender), + 3% DMSO (cryoprotectant), stored at . Group E:Androhep (extender), + 3% ethylene glycol (cryoprotectant), stored at . In group A, the sperm's motility was reduced. On day one the sperm's motility was () and day 5 the motility was (). In group B, C and D the sperm's motility were reduced to 0 on day 5. In group E the sperm's percentage of motility decreased. On day one the sperm's motility was () and day 5 the motility was (). When comparing cryoprotectant in samples of boar sperm there is a slight improvement in the results when the use of Androhep Lite (extender), + 3% ethylene glycol (cryoprotectant), stored at are used. Based on these results, ethylene glycol can protect sperm from heat shock at , but not satisfactory level. However, it showed the possibilities of liquid semen preservation at by using cryoprotectant.

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ( for 20 sec, 45 sec and for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : , NAI : ), TLE (, ) extender significantly(p<0.05) increased than that in LEY (, ) extender thawed at for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE (, ) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE () while that in LEY () is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

넓은 범위의 염분에 대한 망둑어 3종의 적응성을 파악하기 위하여 이들의 생존 및 성장에 대한 염분농도의 영향을 검토하였다. 두줄망둑은 0~33.6‰의 염분범위에서 유의한 생존율 차이가 없었고, 점망둑의 생존율은 염분농도 13.4‰ 이상에서 90% 이상을 나타내었으며, 3.4‰ 이하에서는 80% 이하로 유의하게 감소하였다. 또한, 날개망둑의 생존율은 염분농도 3.4‰ 이상에서 90% 이상을 나타내었으나, 담수에서는 87.5%로 유의한 감소가 관찰되었다. 두줄망둑은 염분농도 13.4 및 20.2‰에서 가장 높은 일일성장률은 나타냈고, 담수에서 유의한 감소가 관찰되었다. 점망둑의 일일성장률은 염분농도 20.2~33.6 ‰ 이상에서 가장 높았고, 10.1‰ 이하에서 가장 낮아 이들 사이의 유의한 차이를 나타냈다. 날개망둑의 일일성장률은 염분농도 10.1~27.4‰에서 가장 높았으나, 담수에서 유의한 감소가 나타났다. 이상의 결과는 3종의 망둑어, 특히 점망둑은 낮은 염분에 노출되었을 때, 그들의 생존 및 성장에 영향을 받는다는 것을 의미하고 있어 이들의 분포 및 개체수에 잠재적인 영향을 받을 수 있을 것으로 예상된다.

In this study, the contamination levels of total aerobic bacteria, E. coli, total coliform and S. aureus of seasoned dried fishes (SDF) in Korea were investigated. A total of 81 SDF samples were purchased randomly from 28 stores. Contamination range of total aerobic bacteria, total coliform and S. aureus were 150~1,700,000, 10~31,000and 10~220 CFU/g, respectively. E. coli was detected in only one samples in the qualitative test. We have analyzed quantitatively Staphylococcal enterotoxins (SE-type A, C and D) produced by S. aureus contaminated in SDF using a TECRA kit and standard curve. The curve equation was Y = 0.1499 * X + 0.1183 and maximum amount of SEs in SDF was 0.71 ng/ml. Reduction speed of S. aureus in SDF stored at 37 ℃ was the highest among the samples stored for 8 days at different temperature of 7, 18 and 37℃ . On the basis of the results, SDF in Korea can be contaminated by a variety of pathogenic bacteria. Therefore, precautionary measures are necessary for consumer protection, including the improvement of sanitary conditions in the processing plants in Korea.

본 연구는 누에의 식엽성 곤충을 대체할 실험곤충으로서의 유용성과 자외선이 한국산 곤충병원성선충 (Heterorhabditis sp. 202 strain, Heterorhabditis sp. Gyeongsan strain, Steinernema sp. 223 strain, S. carpocapsae Pocheon strain, S. glaseri Dongrae strain, S. longicaudum Nonsan strain) 의 병원성과 생존에 미치는 영향을 알아보기 위하여 수행하였다. 누에에 대한 곤충병원성선충의 병원성을 조사한 결과 누에의 령기나 곤충병원성선충의 계통이나 종에 따라 상이한 병원성을 나타내었는데 5령충에 대해서는 Steinernematidae선충의 병원성이 높았고, 1령충에 대해서는 Heterorhabditidae선충의 병원성이 높았다. UV-C는 곤충병원성선충에 유해하여 노출 60분후에는 모든 선충이 치사되었다. UV-C 조사는 곤충병원성선충의 병원성에도 영향을 미쳐 노출 10분 후에도 누에나방 3령충에 대하여 6.7% 이하의 보정사충율을 나타내었다. 야외 뽕나무밭(UV량: 2.3mW/㎠)에서 곤충병원성선충, S. carpocapsae Pocheon strain을 엽면 살포 4시간 후 채취한 뽕나무 잎을 급상한 누에는 치사되지 않았다. 뽕잎에서 4.0mW/㎠의 UV 1시간 노출은 곤충병원성선충, S. carpocapsae Pocheon strain의 누에에 대한 병원성에 차이를 보이지 않았다.

 ,  , The effects of gamma irradiation on the survival, development, fecundity, and sex ratio of the multicolored Asian ladybird beetle (Harmonia axyridis) were investigated. A dose of more than 50 Gy dose of irradiation on eggs completely inhibited hatch, and 1st and 3rd instar larva irradiated with over 40 Gy almost died. Adverse effects of gamma irradiation over 70 Gy were also observed in the survival and development of pupae and adults. tail indicating DNA damage was showed in survival individuals exposed to gamma irradiation.Although irradiated individuals usually survived, they had longer periods of larval and pupal development than untreated individuals. Slight female-biased sex ratios were observed in some treatments. However, sex ratio differences soon equilibrated. Generally, inherited harmful effects of gamma irradiation were observed in F1 and F2 generations. Comet tail indicating DNA damage was showed in survival individuals exposed to gamma irradiation.

식물의 초기 생활사가 종자크기에 따라 어떤 영향을 받는지를 알아보기 위하여 우리나라 산림군락에서 상수리나무, 굴참나무, 갈참나무, 졸참나무 참나무 4종의 열매를 3등급의 크기(대, 중, 소)로 구분하고, 그에 따른 4종의 발아율, 겨울철 생존율과 초기 생장율을 측정하였다. 평균 발아율은 종자가 큰 상수리나무와 굴참나무에서 높았고, 상대적으로 크기가 작은 갈참나무와 졸참나무에서 낮았다. 그러나 종 내 종자크기에 따른 발아율은 굴참나무에서만 종자가 작을수록

돼지 정액 채취과정은 무균적으로 진행되기가 어려우므로 채취된 정액의 세균오염 (bacteriospermia)은 일반적인 것으로 알려져 있다. 돼지 정액에서 주로 분리되는 오염 세 균은 그람음성균이고, 장내세균과(Enterobacteriaceae)에 속하는 것으로 조사되고 있다. 정액 내 세균오염은 오염 정도에 따라 정액의 품질과 수명에 부정적인 영향을 미치는 것 으로 보고되고 있다. 본 실험에서는 돼지 정액에서 순수분리한 대장균을 실험적으로 농 도별로 오염시켜 돼지 정액의 활력, 생존율 및 pH에 대한 대장균 오염의 영향을 확인코 자 하였다. 정자 농도와 세균오염 정도를 조절하기 위하여 항생제가 첨가되지 않은 시판 돼지 정 액 희석제(Seminark Pro)를 이용하였고 돼지 정자수 대비 대장균수가 4,000 : 1(T1), 400 : 1(T2), 40 : 1(T3), 4 : 1(T4)이 되도록 액상정액 시료를 제조하였다. 모든 시료는 17℃ 저온배양기에 보존하면서 유세포분석기(flow cytometer)를 이용하여 0일차, 1일차, 3일차 및 5일차에 각 시료별 정자 활력(미토콘드리아 함량 측정)과 생존율(Live/DeadⓇ 염색)을 측정하였고, 산도측정기(pH meter)로 정액 pH를 측정하였다. 정자의 활력은 T3, T4에서 0일차(17℃, 4시간 배양)부터 대조군(C)에 비하여 유의성 있 게 감소하였고, 모든 시료에서 3일차 이후부터 활력이 감소(p<0.05)하였다. 정자 생존율 의 경우에도 T3, T4에서 0일차부터 C에 비하여 유의성 있게 감소하였고, 모든 시료에서 3일차부터 생존율이 감소(p<0.05)하였다. 보존일 경과에 따른 정액의 pH 변화의 경우, C, T1 및 T2에서는 0일차에 pH 7.00 정도에서 5일차(p<0.05)에 pH 7.10 이상으로 높아진 반면, T3에서는 3일차(p<0.05)부터 pH 6.90 이하로 낮아져 5일차에는 pH 6.86이었고, T4 에서는 1일차(p<0.05)부터 pH 6.86 이하로 낮아져 3일차(p<0.05)와 5일차(p<0.05)에는 pH 6.65 이하로 낮아졌다. 본 결과로부터 돼지 정자수 대비 대장균수가 40 : 1(20×106 sperm cells/ml : 5×105 cfu/ ml) 이상으로 오염되었을 경우 오염 당일부터 5일차까지 대조군에 비하여 정자의 활력과 생존율이 유의성 있게 감소되었으며 40 : 1의 경우는 3일차부터, 4 : 1의 경우는 1일차부 터 정자의 사멸 혹은 세균오염에 의한 산패로 정액의 pH가 낮아지는 경향을 확인할 수 있었다. 이는 정액 내 세균오염이 정액의 품질과 수명에 부정적인 영향을 미친다는 보고 와 일치하며 위생적인 정액 채취를 위한 노력과 고품질 정액 공급을 위한 정기적인 모니 터링 검사 및 보존액 내 유효한 항생제 첨가가 중요할 것으로 사료된다.

한우 인공수정용 액체질소 공급이 월1회로 정액과 같이 공급되고 있어 특히 고온 계절 에는 질소 탱크에 액체질소가 항상 일정하게 유지되고 있지 않은 실정이며 유통과정 중 에 외기에 노출되는 기회가 많이 있음으로써 동결정액의 품질저하가 쉽게 발생된다. 생산 후 유통과정에 발생되는 동결정액의 품질 평가 시기존방법의 단점인 스트로 폐기와 장비 구입비 및 검사 소요시간을 보완하고 동결정액의 품질 변화가 일어나는 ‒160℃ ‒35℃ 사이의 품질을 조사할 수 있게 하여 유통과정상의 소동결정액의 품질저하를 방지 하고 품질 평가를 실시하기위하여 제작하여 조사 하였다. 가축 인공 수정소에서 소동결 정액이 ‒100℃에 노출되는 기회가 56.5%, ‒35℃에 노출은 8.7%가 발생되었으며, 이러한 결과는 동결정액의 생존성을 ‒100℃에서 노출 될 때는 26.9%, ‒35℃에서 노출될 때는 ‒54.4%나 저하됨으로써 농가 인공수정 수태율 저하를 일으키는 주요한 원인이 되고 있 는 것으로 조사되었다.

The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into LN2. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol (72.5±5.00%, 54.88±0.66% and 46.00±2.40%; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol (34.69±4.64% vs 46.00±2.40%; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: 55.81±2.94, 55.19±3.34 vs 47.94±3.48%; p<0.05 and membrane integrity: 44.94±3.51, 46.06±2.25 vs 40.38±1.03%; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

This study was conducted to establish a freezing method of miniature pig spermatozoa. The semen ejaculated from PWG M-type miniature pig was collected by gloved-hand method. The semen was diluted with same volume extender (m-Modena B). The frozen solution used frozen solution of four different (LEY, TCG, BF-5 and m-Modena+egg yolk) for find optimal frozen solution in miniature pig sperm. The diluted semen for frozen rate assay was added to LEY solution (solution Ⅰ: 11% lactose+egg yolk; solution Ⅱ: solutionⅠ+glycerol+OEP), and frozen depending on freezing rate by the three different freezing methods (A: until 5℃ for 1 hrs, holding at —102℃ for 10 min; B: until 5℃ for 2 hrs, holding at —102℃ for 10 min; C: until 5℃ for 3 hrs, holding at —80 and —102℃ for 10 min). Semen cooled until 5℃ was added with glycerol 1, 3 and 5%, and take a equilibrium time for 0, 10 and 30min. Frozen-thawed sperm were evaluated for viability, acrosomal status and morphological abnormality. The results of frozen-thawed sperm ability by frozen solution, viability was higher in LEY solution compared to other three different frozen solution. AR pattern of LEY solution were lower than other three different frozen solution. The results of freezing rate, viability was higher in B method compared to other methods (p<0.05). Acrosomal statute was intacted in A and B methods than C method. The experiment for glycerol condition was showed that sperm viability was higher in extender with 1% and 3% glycerol and equilibrium time of 0 min. The acrosome damage was lower in extender with 1% glycerol and equilibrium time of 10 min than other conditions. In conclusion, the optimal conditions for cryopreservation of miniature pig spermatozoa obtained in LEY frozen solution, cooling rate of 1～2 hrs, 1～3% glycerol concentrations and glycerol equilibrium time of 0～10 min.

The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted (30×106 spermatozoa/ml) in different extenders. Semen was stored at 17℃ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection. The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significantly different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.

포유동물 수정란의 동결보존기술은 최근 기후 변화에 따른 생물종 다양성을 보존하기 위해서 중요하게 여겨지는 연구 분야이다. 따라서 멸실 위험에 처한 동물의 개량과 증식, 보존과 복원 및 생명공학의 분야에 이르기까지 응용 기술은 다양하게 이용되어진다. 본 연구에서는 한우 수정란의 동결 후 생존성 향상을 위해서 동결 방법에 따른 체내 외수정한의 내동성을 조사하였다. 완만동결에 따른 체내 외수정란의 동결 융해 후 수정란의 재확장률은 89.6%와 81.5% 그리

This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN. The presented straws were examined the viability and motility after thawed at 37 water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

Spermatozoa sorted by flow cytometry have been successfully used to produce offspring in domestic animals and are commercially available for cattle. Also sheath fluid is the important environment for viability of sex-sorted sperm in flow cytometry. The aim of this study was to investigate whether or not HEPES (N-2-hydroxyethylpiperazine-N'-2-Ethanesulfonic acid) has any effect on the viability in sex-sorted Hanwoo (Korean native cattle) sperm. In this study, the semen was collected from Hanwoo of Hoengseong Livestock Cooperation by artificial vagina method then pooled and subjected to cryopreservation in straws. Sperm were cultured for 0, 30, 60 and 120 min with 0, 2.5, 5, 7.5 and 10 mM of HEPES added to the sheath fluid and incubated at 4, 20 and 38, respectively. For the cytometric analysis the frozen-thawed semen was extended with 5 mM HEPES extender to final concentration ( spermatozoa) at 4, 20 and 37. Sperm viability was assessed with SYBR-14 and propidium iodide (PI) staining. This study shows that the viability of sperm was decreased with prolongation of incubation time in all of test. But the viability of sperm which were treated with 38 was gently decreased than that of treated with other temperature. The viability of the control was sharply decreased (p<0.05) than all of the HEPES treatment group at 60 to 120 min in 38. X-sexed sperm was more sensitive than Y-sexed sperm to temperature during f10w cytometry (p<0.05). In conclusion, the results of this study suggest that the sheath fluid with 5 mM HEPES has effect on maintenance of viability after sperm sexing at 37 in Hanwoo.

An Unmanned Aerial Vehicle (UAV) is a powered pilotless aircraft, which is controlled remotely or autonomously. UAVs are an attractive alternative for many scientific and military organizations. UAVs can perform operations that are considered to be risky

This research is to select a path planning algorithm to maximize survivability for Unmanned Aerial Vehicle(UAV). An UAV is a powered pilotless aircraft, which is controlled remotely or autonomously. UAVs are currently employed in many military missions(surveillance, reconnaissance, communication relay, targeting, strike etc.) and a number of civilian applications(communication service, broadcast service, traffic control support, monitoring, measurement etc.). In this research, a mathematical programming model is suggested by using MRPP(Most Reliable Path Problem) and verified by using ILOG CPLEX. A path planning algorithm for UAV is selected by comparing of SPP(Shortest Path Problem) algorithms which transfer MRPP into SPP.