This study was conducted to comparing on vitrification of mouse oocytes and embryos using CPS, conventional straws and CPS by evaluating in morphological survival for oocytes, and embryonic cleavages and blastocyst formation for embryos. The morphological survival in vitro after thawing of vitrified oocytes using CPS (75%) and conventional straws (72%) were significantly higher (p<0.05) than that using OPS (68%). The blastocyst formation rates of vitrified embryos using CPS (48.6%) and unfrozen control embryos (56.0%) were significantly higher (p<0.05) than those of conventional straws (43.4%) and OPS (37.7%). The rates of morula formation were also higher to control, CPS, conventional straws and OPS in orderly. These results show that CPS has the advantages of achieving a high survival and safety preservation.
To develop an effective vitrification method, we examined the use of a conventional straw as vessel fur vitrification of mouse oocytes, and to compare the post-thaw survival and chromosome configuration of these oocytes with those vitrified in grids. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded into straws and onto eletron microscopic copper grid fur storing in liquid nitrogen. Intact vitrified and thawed oocytes were karyotying for chromosome. The rates of post-thawed survival were 88.5% in vitrified oocytes with straws, and 83% in vitrified ooctyes with grids. Vitrified and thawed oocytes with straws and grids were increased chromosomal abnormality (31.4% and 30.9%) compared with fresh oocytes (17.8%). The conventional straws can be used as vessel for vitrification to prevent of inflection in liquid nitrogen.
Park, Young-Hwan;Chang, Hak-Gil;Jung, Chung-Sam;Kim, Dong-Soo
The Korean Journal of Mycology
/
v.2
no.1
/
pp.21-24
/
1974
Present experiments were conducted to determine the possibility of cultivation of tropical and subtropical saprophytic fungus, Volvariella volvacea (Bull. ex Fr.) Sing. under Korean climatic conditions and the-possibility of applying the method of indoor cultivation which was modified from the cultivation method of the common mushroom, Agaricus bisporus (Lange) Sing.. The results obtained were as follows: 1. In conventional outdoor cultivation, the adequate cultivation period of this fungus was from around the end of June to the end of August. 2. When rice straws submerged in water for 5 days were peak-heated for 6 days in a mush room house, the yield of the fungus was higher than that obtained from the rice straws peak-heated immediately after outdoor composting. 3. The highest yield, 20.49kg per 100kg rice straws, was obtained from the water-soaked and peak-heated rice straws which were cased with a 2 cm layer of loam at 7 days after spawning.
No-till direct seeding cultivation of rice has major advantages such as saving of labor and cost by eliminating tillage, preparation of seed bed and trans-planting procedure compared to the conventional transplanting cultivation. A field experiment was conducted to evaluate the effects of straw treatment and nitrogen levels on the rice growth in no-till direct-seeding cultivation. Rice straw, vetch straw, and the mixture of both of the straws were mulched on the surface of soil before seeding while 4 levels of nitrogen fertilizer, 0, 7, 9, and 11 kkg/10a respectively, were applied at 3 split times, 3-weeks after sowing, 5-weeks after sowing and the panicle initiation stage. Mulching of vetch straw significantly reduced seedling establishment of rice which may be attributed to low oxidation-reduction potential of soil by vetch mulching treatment. Vetch straw increased the concentration of soil ammonium leading to an extension of the greenish leaf to panicle initiation stage. Agronomic nitrogen use efficiency (AD $E_{N}$) in heavy-mixed straw mulching plots was lower than other treatments. Grain yield and AU $E_{N}$ in the vetch treatment were less affected by fertilized N levels. Conclusively, it is suggested that heavy straw mulching was not efficient for rice seedling establishment and nitrogen usage.e.
The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.
The cryopreservation of Hanwoo embryos has become an integral part of assisted reproduction in animal. The objective of this study was to assess the effect of The objectives of this study were: (1) to evaluate the influence of bovine embryo developmental stage on in vitro embryo development after freezing, (2) to study the efficiency compared with conventional freezed embryos at different embryo source. For conventional slow-freezing, day 7 or 8 expanded blastocysts were collected. The standard freezing medium was 1.8 M ethylene glycol (EG). Embryos were equilibrated in 1.8 Methylene glycol(EG) with 0.1 M sucrose in Dulbecco's phosphate-buffered saline (D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25 ml-straw and placed directly into cooling chamber of programmable freezer precooled to $-7^{\circ}C$, after 2 min, the straw was seeded, maintained at $-7^{\circ}C$ for 8 min, and then cooled to $-35^{\circ}C$ at $0.3^{\circ}C$/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 see and exposed to $37^{\circ}C$ water for 20 sec. Straws were then removed from $37^{\circ}C$ water. Rates of blastocyst survive and hatched were evaluated at 12 to 48h post-warming. The re-expansion and hatched rates of morula embryos were significantly lower than those obtained for blastocysts and expansion blastocysts (31.6%, 10.5% vs, 68.9%, 22.2% vs, 73.7%, 53.6%, respectively). No differences in re-expansion rates were found between in vivo and in vitro blastocysts. whereas hatched rates was significantly higher (51.2%) in vivo compared with in vitro embryos (18.6%). in conclusion, demonstrate that conventional freezing can be used successfully in cryopreservation of in vitro and in vivo bovine embryos, and that it might be considered for use in commercial programs and embryo preservation.
This study was conducted to investigate the effects of vitrification solution and developmental stage on the survival rate of vitrified-thawed human blastocyst embryos. Human blastocyst embryos were cryopreserved by vitrification using EFS and GE solution, and their survival rates were examined after thawing and further culture. EFS solution was consisted of 40% ethylene glycol, 18% Ficoll 70 and 0.3M sucrose. GE solution was consisted of 25% glycerol and 25% ethylene glycol. Embryos were exposed to EFS and GE solution by 2 steps and 3 steps, respectively, and plunged into liquid nitrogen after loading into 0.25ml plastic straws. Blastocysts were classified into 4 groups in accordance with their developmental stage: into 1) EEB, 2) MEB and 3) EdB, of blastocysts developed on day 5, and 4) 6d-Bla(the blastocysts which formed on day 6). The blastocysts at each stage were vitrified by GE solution and cryopreserved in LN2. After thawing them, we examined their survival rates, respectively. The resulted of this study were as follows: 1. The survival rate of blastocysts vitrified by GE solution was 64.4%, significantly higher than that (5.7%) vitrified by EFS solution (P<0.001). 2. When the blastocysts were vitrified by GE solution according to each developmental stage, the survival rates of EEB, MEB, EdB and 6d-Bla were 65.9%, 65.9%, 73.2% and 58.1%, respectively. In conclusion, the cryopreservation of human blastocysts by vitrification is likely to have a marked advantage in terms of cost, work and time as compared to the conventional slow freezing in IVF-ET programs.
Real time and robust algorithm to extract the features of watermelon was developed from the remotely transmitted image of the watermelon. Features of the watermelon at the cultivation site such as size and shape including position are crucial to the successful tole-robotic operation and development of the cultivation data base. Algorithm was developed based on the concept of task sharing between the computer and the operator utilizing man-computer interface. Task sharing was performed based on the functional characteristics of human and computer. Identifying watermelon from the image transmitted from the cultivation site is very difficult because of the variable light condition and the complex image contents such as soil, mulching vinyl, straws on the ground, irregular leaves and stems. Utilizing operator's teaching through the touch screen mounted on the image monitor, the complex time consuming image processing process and instability of processing results in the watermelon identification has been avoided. Color and brightness characteristics were analyzed from the image area specified by the operator's teaching. Watermelon segmentation was performed using the brightness and color distribution of the specified imae processing area. Modified general Hough transform was developed to extract the shape, major and minor axes, and the position, of the watermelon. It took less than 100 msec of the image processing time, and was a lot faster than conventional approach. The proposed method showed the robustness and practicability in identifying watermelon from the wireless transmitted color image of the cultivation site.
Kim, Young Il;Lee, Sang Moo;Park, Keun Kyu;Kwak, Wan Sup
Journal of The Korean Society of Grassland and Forage Science
/
v.33
no.4
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pp.290-297
/
2013
This study was carried out to investigate the effects of a by-product feed-based silage (BF silage) feeding on behavior patterns of growing Hanwoo steers. A total of 10 Hanwoo steers (11 months old, 302kg of body weight) were assigned to 2 dietary treatments: the control (concentrate mix + free access to rice straw), and the treatment (concentrate mix + free access to BF silage). The behavior patterns were observed for 48 hours. The intakes of dry matter (DM) and neutral detergent fiber (NDF) of the treatment group were higher than those of the control group. Eating time, ruminating time and resting time were not different between the control and treatment. But, the intake time per kg DM was higher for the control than treatment. The number of bolus, total chewing frequency, number of ruminating per bolus and number of bolus per minute were not different between the control and treatment. But the chewing frequency per bolus was higher in the treatment than control (p<0.05), and feed value index was lower in treatment than control (p<0.05). Frequencies of drinking and defecating were not different between the two groups, but the frequency of urinating was higher for the treatment (p<0.05) than control. Eating rate, ruminating efficiency and chewing efficiency were much higher in the treatment group than control (p<0.05). These results indicate that the replacement of conventional rice straws with the BF silage (physically effective NDF, about 25%) did not affect the ruminating behaviors of Hanwoo steers significantly.
The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 7°C, after 2 min, the straw was seeded, maintained at 7°C for 8 min, and then cooled to 35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.
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