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.
The objective of this study was carried out to evaluate the efficiency of sperm collection methods on the post-thaw viability of cat semen. The cat semen was collected by artificial virginal (AV) and electronic ejaculate (EE) methods. The composition of semen extender was consisted of Tris-buffer supplemented with 20% egg yolk and 1% P/S antibiotics in Ext I, and more added 8% glycerol, 1.0% Equex STM paste of total volume in Ext II. The collected semen was adjusted the concentration and then diluted in Ext I for optimal concentration. The diluted semen was cooling to 5℃ temperature in refrigerator for at least 2 hrs and then diluted stepwise with Ext II for at least 1 hrs. After an equilibration for 1 hrs, the cooled semen was packaged in 0.5 ml straw and then freezing on the LN2 vapor over 5 cm above from LN2 and then immersed directly in LN2 for cryopreservation. The frozen semen was thawed in 38℃ water for 15 sec and then evaluated the motility, viability, and morphology. Post-thaw semen were calculated the motility by SMI (sperm motility index). The live-dead sperm was evaluated by Eosin-B and morphological evaluation was by Diff-quik kit staining. The post-thaw concentration (89×106 /ml vs. 128×106 /ml), viability (22.6±10.6% vs. 37.1±26.1%), morphological normality (27.0±50.2% vs. 45.6±123.0%) of EE and AV groups were not significant different, but the post-thaw motility was significant lower in EE than that in AV group (53.1±3.6 vs. 73.6±5.7) (p<0.05). In conclusion, semen collection methods did not significant different between EE and AV groups except of post-thaw motility and so both semen collection methods could be applied in feline semen collection methods.
Arcobacter butzleri is one of the aerotolerant Campylobacter species which cause persistent diarrhea,abdominal pain, nausea and vomiting in human. The aim of this study was to determine the growth and survival of A. butzleri under the heat treatment or freezing storage condition. In heat treatment, two Korean isolates of A. butzleri were treated at 40 to 80℃ for various times. In freeze treatment, two Korean isolates of A. butzleri were kept at 4, −20 and −70℃ from 1 to 15 d. The survivability of the A. butzleri Korean isolates significantly decreased at higher than 60℃ heating condition but it didn't show any significant difference in 40℃ treated group. Under the cold stress condition, survivability of A. butzleri were significantly decreased at −20℃ storage. Like other foodborne pathogens,survivability of A. butzleri was controlled by heat and freezing treatment.
NAC는 GSH의 전구물질로, thiol기를 포함하는 항산화제 중 하나로 잘 알려져 있으며, 방사선 조사 시 발생하는 생체 내 영향을 감소시켜 생체 손상의 방호 및 회복에 도움을 주는 방사선 방어제로 이용된다. S. cerevisiae에서 항산화제 NAC를 전처리 함에 따라 이온화 방사선 조사에 따른 효모의 세포사멸 방어효과 및 superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx)와 같은
Recent competitive and technological changes during the past decade have accelerated the need for better capital recovery methods. Competition and technology have together shortened the expected lives of property which could not have been forecasted sever
This study was conducted to investigate the survival rate of frozen-thawed spermatozoa in equine by glycerol concentration and freezing speed. Two stallions (1 Thoroughbred-13 year old and 1 Arab-7 year old) bred in Korea Racing authority was examined for 1 times in a couple of weeks. Semen was collected by condom method standing heated mare and were centrifuged 650 g for 15 min. and isolated the seminal plasma. Thick fraction of semen was diluted EDTA-Lactose-egg yolk diluents to 1:1 and contained in 0.5 ml straw as 6~14×107cells/ml. Final concentrations of glycerol were 3, 5 and 7% in cryopreseved diluents and added 4 times for 2 hours equilibration. For the freezing, equilibrated straws were located 3 or 5 cm above LN2 gas for 5 or 10 min. Survival rates of pre-frozen sperm were 65.0±13.2%, 68.3±10.4%, 66.7±11.5% and post-frozen were 53.3±23.1%, 45.0±15.0%, 50.0±18.0% in 3, 5, 7% glycerol concentration, respectively. There was no difference between glycerol concentrations. Survival rates of frozen-thawed sperm on freezing speed were 36.7±10.4%, 40.0±7.1%, 30.0±13.2% at 3 cm-5 min and 33.3±11.5%, 31.7± 2.9%, 21.7±10.4% at 3 cm-10 min in 3, 5, 7% glycerol concentration, respectively. Survival rates of frozen-thawed sperm on freezing speed were 43.3±15.3%, 32.0±17.9%, 22.3±15.7% at 5cm-5 min and were 47.5±15.0%, 43.3±12.6%, 48.3±15.3% at 5cm-10 min in 3, 5, 7% glycerol concentration, respectively. There were significantly different between groups (p<0.05). These results suggest that glycerol concentration did not affect cryopreservation of stallion semen within 3~7% but freezing speed affects. In our experiment, the best cryopreservation condition was at 5 cm above LN2 gas for 10 min for pre-freezing and 7% of glycerol concentration. These results lead to commercial AI with frozen-thawed stallion semen.