In this study, an experiment was conducted in order to determine what cryopreservatives (CPVs) were more effective in supporting the motility and viability of sperm from experimental animals. The sperm of mice, rats, beagle dogs, and rabbits were frozen using different CPVs, including DMSO, TYB, and Sperm CryoProtec. The results from freezing the sperm of each laboratory animal in Sperm CryoProtec showed a high level of sperm motility and viability in sperm samples from mice, rats, and beagle dogs melted at the end of the first week. For rabbits, a high level of motility was observed in sperm thawed during the first week, whereas a high level of viability was observed in sperm thawed during the second week. The results of analysis of sperm motility and viability using different CPVs according to laboratory animals showed a significantly higher level of sperm motility (26.28%) and viability (36.20%) for mice in Sperm CryoProtec and the lowest levels of motility and viability were observed in DMSO (p < 0.05). Significantly higher levels of motility (27.94%) and viability (37.94%) were observed for rats in Sperm CryoProtec compared with TYB, which showed the lowest levels of motility and viability (p < 0.05). The study findings described above suggest that the selection of appropriate cryopreservatives is required for each experimental animal. This is because there are differences in the levels of sperm motility and viability of experimental animals depending on the CPVs that are typically used for freezing human sperm, including Sperm CryoProtec and TYB.

Reactive oxygen species (ROS) production and F-actin cytoskeleton dynamics play important roles in the survival rate of blastocysts after the vitrifiedwarming process. However, the protective effects of Mito-TEMPO against cryo-injury and viability through F-actin aggregation and mitochondrial-specific ROS production in vitrificated-warmed bovine embryos have not been investigated. The aims of the present study were to: (1) determine the effects of Mito-TEMPO on embryonic developmental competence and quality by F-actin stabilization during in vitro culturing (IVC), and (2) confirm the effects of Mito-TEMPO through F-actin structure on the cryotolerance of vitrification-warming in Mito-TEMPO exposed in vitro production (IVP) of bovine blastocysts. Bovine zygotes were cultured with 0.1 μM Mito-TEMPO treatment for 2 days of IVC. Mito-TEMPO (0.1 μM) exposed bovine embryos slightly improved in blastocyst developmental rates compared to the non-treated group. Moreover, the viability of vitrified-warmed blastocysts from Mito-TEMPO treated embryos significantly increased (p < 0.05, non-treated group: 66.7 ± 3.2% vs Mito-TEMPO treated group: 79.2 ± 5.9%; re-expanded at 24 hours). Mito-TEMPO exposed embryos strengthened the F-actin structure and arrangement in the blastocyst after vitrification-warming. Furthermore, the addition of Mito-TEMPO into the IVC medium enhanced embryonic survival and quality through F-actin stabilization after the vitrification-warming procedure. Overall, our results suggest that supplementing the culture with 0.1 μM Mito- TEMPO improves the embryonic quality and cryo-survival of IVP bovine blastocysts.

This research aims to investigate pudding with grain-added yogurt for its quality characteristics and viability during cold storage. The yogurt was fermented until its pH was 5.10±0.05 after inoculating the probiotic strain (Bifidobacterium lactis, BB-12) into the milk base containing grains. The yogurt was added to prepare probiotic puddings. During cold storage of the puddings at 4±1oC for 4 week, the quality characteristics (pH, acidity, texture) and the viability of BB-12 in pudding were determined and compared to control (only milk base). As a result, MR had a significantly lower pH and higher acidity than those of other samples. In texture properties, including hardness, gumminess, and chewiness, MSIR showed the significantly highest value, and the pudding with inulin was significantly higher than rice flour in all textures. For the viability of BB-12, pudding with milk was significantly lower than pudding containing milk and soymilk, suggesting that soymilk helps maintain viability. MR showed significantly higher viability than MI in the milk-based pudding, indicating that rice flour is more effective than inulin. Therefore, the addition of soymilk, inulin, and rice can maintain quality characteristics and viability of BB-12 in the pudding.

This research aimed to investigate the effects of the inoculation method and diverse ingredients on the quality properties and probiotics viability of pudding with milk and/or soymilk. The probiotic strain (Lactobacillus acidophilus, LA-5) was inoculated into the yogurt base and fermented until the pH is 5.10 ± 0.05. The fermented yogurt was added into pudding base (yogurt inoculation), or LA-5 was directly inoculated (direct inoculation). During the storage of the puddings at 4±1oC for 4 wks, the quality characteristics (pH, acidity, texture) and the probiotics viability of pudding were measured and compared. As a result, the puddings with yogurt inoculation showed significantly lower pH and higher acidity than those with direct inoculation. In texture properties, including hardness, chewiness, and gumminess, the addition of rice powder increased those in milk pudding more but the addition of inulin in the milk-soymilk pudding more. The addition of rice powder increased probiotics viability more than inulin in milk and milk-soymilk puddings. Therefore, adding rice and/or inulin can potentially improve the probiotics viability and quality characteristics of pudding.

본 연구는 시중에 유통 중인 유산균 첨가 시료 6개를 7개월 동안 저장하여, 유산균 생균 수의 변화와 우수 균종을 분리 및 분석하고자 하였다. 제품별 저장 기간에 따라 유산균 생균 수의 변화는 MRS agar에서 9.04 Log CFU/g에서 7.20 Log CFU/g, BCP agar에 서 8.96 Log CFU/g에서 7.63 Log CFU/g으로 7개월이 경과한 후에 유의적으로 감소하였다. 식품위생검사기관에서 기간별 시료의 평균 유산균 생균 수는 6.58 Log CFU/g 이상 유지하여, 본 연구 결과와 76.0%–100% 일치하는 경향을 나타냈다. 제품의 유 산균 표시 함량은 2.7×106–6.2×106 CFU/g으로 표시되었으며, 본 연구 결과에서 1.5×107–1.4×108 CFU/g 이상 검출되어 실제 외포장지의 표시 함량은 올바른 표시에 부합되었다. 검출된 유산균 중 우점종(85%)을 최종 선발하여 16S rRNA 유전자 염기서열 분석 결과, Lactiplantibacillus(identity 100%), Streptococcus thermophilus 10%(identity 99%) 및 Weissella confusa 5% (identity 99%)로 동정되었다. 이는 본 연구의 시료에서 검출된 우수 균주로, 위 균주 외에 또 다른 우수한 균주를 선발하여 식품에 접목하는 연구가 필요한 것으로 생각된다. 또한 6개 제품 모두 7개 월 후 유산균 수 함량이 107–108 CFU/g으로 확인되었으며, 실험한 제품류(비스킷류, 샌드류 및 초콜릿류)의 외포장지에 표시된 유산균 함량은 본 실험의 결과보다 적게 표시되었으며, 유통기한(6개월) 동안 유산균의 함량은 유지되었다.

In the present study, we examined the effect of straw size on spermatozoa motility, viability, acrosome integrity, mitochondrial membrane potential, and plasma membrane integrity after freezing-thawing. Hanwoo semen was collected from three bulls and diluted with an animal protein-free extender, divided into two groups, namely, 10 million spermatozoa in 0.25 mL and 20 million spermatozoa in 0.5 mL straw, and cryopreserved. In Experiment 1, the motility and motility parameters of the frozenthawed spermatozoa were evaluated. After freezing-thawing, the spermatozoa motility parameters fast progressive, straight line velocity, and average path velocity were compared between the 0.25 mL straw and 0.5 mL straw groups. They were 35.2 ± 1.0 and 32.3 ± 0.7%, 34.6 ± 0.7 and 31.8 ± 0.5 μm/s, 51.4 ± 1.3 and 47.1 ± 1.1 μm/s, 0.25 mL straw and 0.5 mL straw groups, respectively. In Experiment 2, the viability, acrosome membrane integrity, and mitochondrial membrane potential of the frozen-thawed spermatozoa were assessed. After freezing-thawing, the percentages of spermatozoa with live, intact acrosomes and high mitochondrial membrane potential were compared between the in 0.25 mL straw and 0.5 mL straw groups. They were 48.0 ± 2.6% and 35.6 ± 2.8% between the 0.25 mL straw and 0.5 mL straw groups. In Experiment 3, the plasma membrane integrity of frozen-thawed spermatozoa was compared. After freezingthawing, the plasma membrane integrity was higher for the in 0.25 mL straw group than the 0.5 mL straw group. They were 62.0 ± 2.2 and 54.1 ± 1.3% between the 0.25 mL straw and 0.5 mL straw groups. In conclusion, our results suggest that freezing semen in 0.25 mL straw improves the relative motility, viability, and acrosomal, mitochondrial membrane potential, and plasma membrane integrity of Hanwoo bull spermatozoa.

Since the first detection of the African swine fever (ASF) virus in the Republic of Korea in 2019, the Korean government has applied interventions, including fencing, increasing the biosecurity level at domestic pig farms, and the capture-and-removal of wild boars. In particular, wild boars are an important risk factor for ASF control because they can spread disease among susceptible animals, such as wild boars or domestic pigs. A capture-and-removal method aims to reduce the likelihood of ASF transmission from wild boars to domestic boars or among wild boars by decreasing the number of susceptible wild boars. This study estimated the required number of wild boars captured and removed for ASF control using population viability analysis. Population factors, such as a life span, sex ratio, or an inbreeding depression with different capture-and-removal proportions of wild boars, were included in the analysis. Ten scenarios with different capture-and-removal proportions of wild boars and different periods of culling were considered. According to the results, a method in which 75% of wild boars are captured-and-removed for at least three years showed long-term effectiveness for more than ten years. The current ASF control method, in which 33% of wild boars are captured-and-removed, decreased the number of wild boars for three years, after which the wild boar population increased to more than its initial number. Given the limited human and material resources for controlling ASF in the Republic of Korea, it is recommended that resources be prioritized to increase the capture-and-removal proportion of wild boars to take full advantage of the ASF-control effectiveness.

Short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate are secondary metabolites produced by anaerobic fermentation of dietary fibers in the intestine. Intestinal SCFAs exert various beneficial effects on intestinal homeostasis, including energy metabolism, autophagy, cell proliferation, immune reaction, and inflammation, whereas contradictory roles of SCFAs in the oral cavity have been reported. Herein, we found that low and high concentrations of SCFAs induce differential regulation of intracellular Ca2+ mobilization and expression of pro-inflammatory cytokines, such as interleukin (IL)-6 and IL-8, respectively, in gingival fibroblast cells. Additionally, cell viability was found to be differentially regulated in response to low and high concentrations of SCFAs. These findings demonstrate that the physiological functions of SCFAs in various cellular responses are more likely dependent on their local concentration.

여주는 열대와 아열대 지역에서 재배되는 1년생 박과 채소로 혈당조절 등의 기능성이 알려지면서 재배면적이 증가하고 있다. 그러나 여주는 자웅동주이화 식물로 인공수분이 필요하고 온도나 일장에 민감하게 반응하는 개화특성 때문에 안정적인 수량 확보가 어려운 경향이 있다. 평균적으로 암꽃과 수꽃의 착생 비율은 1:25 정도이나 생육 초기 단일조건에서는 암꽃 발생이 많아 인공수분이 어렵다. 이 연구는 암꽃 비율이 높아지는 시기에 환경 조건과 관계없이 화분을 안정적으로 공급하고자 화분 활력 및 저장성 검정에 관한 연구를 수행하였다. 화분을 채취하여 상온(25℃), 냉장(4℃), 냉동(-20℃), 초저온냉동(-70℃)조건에서 180일간 저장하여 발아율을 조사한 결과, 상온에서는 저장 3일 경과 후 급격히 감소하였고 초저온냉동저장(-70℃)에서 발아율이 30%로 가장 높게 나타났다. 또한, 화분의 활력도 발아율과 같이 저장온도가 낮을수록 높은 경향을 보였으며, 초저온냉동저장(-70℃) 화분은 69%로 개화당일 화분의 74%와 유사한 수준이었다. 저장온도를 달리하여 저장한 화분으로 인공수분한 결과 결실률도 저장온도가 낮을수록 높게 나타났고, 상품과 평균과중은 처리 간에 큰 차이는 없었다. 이는 저장온도가 화분의 생존율과 결실률에 영향을 미치는 것으로 판단되어 여주의 안정적인 수량 확보를 위해서는 효과적인 화분 공급 체계가 필요할 것으로 생각된다.

The purpose of this study was to evaluate the effect of addition of ethylene glycol, glycerol and sucrose to TCG (Tris, Citric Acid, Glucose, Egg Yolk) and DMSO Frozen. The extender containing Egg yolk concentration (10%, 20%) affects viability and acrosome morphology of rabbit sperm. Sperm viability was then assessed for the freezing extenders TCGD (Tris + Citricacid + Glucose + DMSO), TCGED (Tris + Citricacid + Glucose + Egg yolk + DMSO), TCGGD (Tris + Citricacid + Glucose + Glycerol + DMSO) and TCGSD Tris + Citricacid + Glucose + Sucrose + DMSO) during thawing at 38oC. for 20 seconds, respectively. TCG + 10% egg yolk (viability: 77.0 ± 0.8, NAI: 73.3 ± 0.9) was significantly (sperm viability and normal acrosome interaction (NAI)) higher than TCG + 20% egg yolk (70.7 ± 1.1, 70.0 ± 0.9) in the sperm normalcy analysis according to the yolk concentration. TCGGD (53.4 ± 0.1, 62.3 ± 0.4), TCGSD (61.3 ± 0.0, 67.1 ± 0.1) sperm viability and normal acrosome interaction (NAI) in frozen spermatozoa are TCGD (46.4 ± 2.8 and 56.3 ± 1. 4) and TCGED (23.0 ± 1.1 and 54.6 ± 1.4) extenders was thawed at 38oC for 20 seconds. According to the results from each frozen bulking agent, sperm membrane integrity by hypotonic swelling test (HOST) analysis in TCGGD (59.8 ± 0.7), TCGSD (59.3 ± 0.5) was significantly high compared to other experimental groups (p < 0.05). In conclusion, these results suggested that TCGGD and TCGSD extenders enhance survivability of rabbit sperm after frozen-thawing.

본 연구는 돼지의 정자와 난소내 과립막세포에서 bisphenol S(BPS)가 생존성과 활성산소 생산에 미치는 영향을 알아보고자 연구하였다. 돼지정액은 0, 5μM BPS를 처리하여 3, 6시간동안 배양하였다. 정자의 생존성은 SYBR14/PI를 이중 염색하여 분석하였으며, 활성산소의 생산을 측정하였다. 또한, BPS(0, 5, 10, 20μM)를 과립막세포에 처리하여 24, 48, 72시간동안 처리하였다. 처리 후, 세포의 생존율과 활성산소 생산(단, 5μM BPS)을 측정하였다. 그 결과, 돼지에서 정자의 생존율은 BPS에 의해 감소하였고, 활성산소의 생산은 모든 처리시간에서 증가하였다(p<0.05). 또한 과립막세포의 생존은 BPS에 의해 억제되었고, 활성산소는 유의적으로 증가하였다(p<0.05). 이상의 결과를 토대로, BPS의 노출은 정자의 활성과 번식과 관련된 세포에 나쁜 영향을 미칠 것이다.

We compared the morphology, physiology, and stem-count change of natural versus transplanted endangered orchids (Cypripedium japonicum) undergoing habitat management (repeated removal of competing understory vegetation) during 2009 and 2013 in South Korea. The restored site had lower transmitted light and soil humidity than the natural site. The natural and restored populations differed in leaf morphology and chlorophyll content (natural: total chlorophyll = 1.00 ± 0.04, restored: total chlorophyll = 0.53 ± 0.06). No recruitment occurred during the monitoring period. Population viability tended to decrease in the restored population (λG=0.97,μ=– 0.05, σ2=0.036) and increase in the natural population (λG=1.07,μ = 0.03, σ2=0.075). In conclusion, restored C. japonicum have poor long-term viability compared with plants in the natural habitat, a difference caused mainly by inappropriate transplant-site selection. Repeated removal of competing vegetation differed in its effects on character, abundance, and reproductive properties in both populations. Notably, habitat management increased stem count and flowering rate in natural C. japonicum, but these changes did not cause an increase in fruit-setting rate.

This study was to investigate effect of tunicamycin (TM) on sperm viability, mitochondrial activity and motility in boar semen. Collected sperm were incubated with semen extender containing 0, 1, 2, and 5 μM TM for 3, 6 and 9 h. Sperm viability was analyzed using SYBR14/PI doubling staining, and mitochondrial activity was detected using Rhodamine123/PI staining methods. Sperm viability, mitochondrial activity and motility were measured every 3 h during incubation. In results, boar sperm viability, mitochondrial activity and motility were significantly decreased in 2 and 5 μM TM groups compare to control group at all incubation time (p<0.05). In addition, mitochondrial activity and motility were significantly decreased in 1, 2, and 5 μM TM groups compare to control group at 9 h after incubation (p<0.05). These results suggest that TM can inhibit sperm viability, mitochondrial activity and motility in boar semen, and it may influence on the fertility of sperm.

Antioxidants have been added to cryoprotectant or in vitro culture medium for sperm to reduce the detrimental damage, such as reactive oxygen species. However, curcumin, an antioxidant, shows dual effect on the viability and progressive motility of bovine sperm exposed to hydrogen peroxide. Low concentration of curcumin increases sperm viability and progressive motility, whereas high concentration of curcumin reduces them. This study was performed to identify whether TREK-1 channel is related to low sperm viability and motility induced by high concentration of curcumin. Curcumin reduced TREK-1 channel activity in a dose-dependent manner. TREK-1 channel was expressed in sperm obtained from Korean native bull. Treatment with TREK-1 channel blockers, such as curcumin, fluoxetine, GdCl3, and spadin, significantly reduced sperm viability and motility (p < 0.05). However, TREK-1 channel activators showed different effect; linoleic acid showed an increase in sperm viability and motility, and wogonin did not affect them. These results show that TREK-1 channel is involved in the regulation of sperm viability and motility. In particular, high concentration of curcumin might reduce sperm viability and progressive motility of Korean native bull through blockage of TREK-1 channel.

To preserve genetic materials, cryopreservation of the semen from live animals is the main technique to establish cryo-banking system which could be used for artificial insemination and embryo transfer. However, the population of Korean black goat (KBG) becomes to dwindle in number and is now faced genetic erosion by crossbreeding with non-native breeds in small KBG farms. In this study, simple freezing method was used to preserve frozen semen from KBG using spermatozoa of cauda epididymis (CE) and electro-stimulated semen (ES). The negative effects of seminal plasma on fresh sperm was confirmed using precipitation test of Triladyl egg yolk diluent and sperm viability after thawing was compared between CE and ES spermatozoa. When seminal plasma of fresh ES semen was washed with semen washing media (SWM), the rates of live sperm shown no significant difference between CE and ES spermatozoa before freezing. However, the survival rate of frozen/thawed CE sperm was higher than ES (74.6±10.6% vs 53.8±5.2%) with significant difference (p < 0.05). The results of longevity test on frozen/thawed sperm from CE showed healthier sperm than ES. Therefore, spermatozoa from CE could be used for cryo-banking system in KBG lines. The more studies are needed to increase survival rate of ES semen.

Spermatozoa viability can be assessed by microscopy, flow cytometry, and other methods using fluorescent stain. Flow cytometry can be used to examine the morphological and functional characteristics of spermatozoa in a short time. The purpose of this study was to compare the viability of cryopreserved spermatozoa in Jeju black cattle by two dual fluorescent stain methods. Semen of Jeju black cattle raised in Subtropical Livestock Research Institute, National Institute of Animal Science, RDA were collected with artificial vaginal technique. Sperm was diluted with Triladyl®-egg yolk diluent and then was performed cryopreservation.There was no significant difference in viability of spermatozoa according to the two dual fluorescent stain methods. However, when the distribution of spermatozoa according to the staining method was compared, the spermatozoa group stained with 6-CFDA/PI was more clearly distinguished than the spermatozoa group stained with calcein AM/PI.