• Journal of Embryo Transfer
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• v.33 no.3
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• pp.185-194
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• 2018

The aim of this study was performed to evaluate the effects of ice-binding protein from the arctic yeast Leucosporidium (LeIBP) supplementation on cryopreservation of boar sperm. The collected semen was diluted ($1.5{\times}10^8/ml$) in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h. The cooled semen was then diluted ($1{\times}10^8/ml$) in LeIBP containing LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min. The semen was divided into six experimental groups (control, 0.001, 0.005, 0.01, 0.05 and 0.1 mg/ml of LeIBP). The straws were kept on above the liquid nitrogen ($LN_2$) vapors for 20 minutes and then plunged into $LN_2$. After thawing, computer-assisted sperm analysis was used for sperm motility and flow cytometry was performed to assess the viability, acrosome integrity (FITC-PSA/PI), ROS (DCF/PI), lipid peroxidation (BODIPY C11/PI) and apoptosis (Annexin V/PI), respectively. No significant responses were observed for sperm motility. However, sperm viability was significantly increased on 0.05 and 0.1 mg/ml of LeIBP groups compared to control (P < 0.05). In addition, acrosome integrity was significantly increases LeIBP groups (P < 0.05) and both ROS and lipid peroxidation level were lower in all LeIBP groups than those of control (P < 0.05). On the other hand, a significant higher apoptosis rate was observed in 0.05 and 0.1 mg/ml of LeIBP groups compared to control (P < 0.05). It can be assumed that a supplementation of LeIBP in boar sperm freezing extender is an effective method to increase the sperm qualities after cryopreservation.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.271-274
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• 2010

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 $6{\sim}14{\times}10^7\;cells/ml$. 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 em above $LN_2$ gas for 5 or 10 min. Survival rates of pre-frozen sperm were $65.0{\pm}13.2%$, $68.3{\pm}10.4%$, $66.7{\pm}11.5%$ and post-frozen were $53.3{\pm}23.1%$, $45.0{\pm}15.0%$, $50.0{\pm}18.0%$ in 3, 5, 7% glycerol concentration, respectively. There was no difference between glycerol concentrations. Survival rates of frozen-thawed sperm on freezing speed were $36.7{\pm}10.4%$, $40.0{\pm}7.1%$, $30.0{\pm}13.2%$ at 3 cm-5 min and $33.3{\pm}11.5%$, $31.7{\pm}2.9%$, $21.7{\pm}10.4%$ at 3 cm-10 min in 3, 5, 7% glycerol concentration, respectively. Survival rates of frozen-thawed sperm on freezing speed were $43.3{\pm}15.3%$, $32.0{\pm}17.9%$, $22.3{\pm}15.7%$ at 5cm-5 min and were $47.5{\pm}15.0%$, $43.3{\pm}12.6%$, $48.3{\pm}15.3%$ 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 $LN_2$ gas for 10 min for pre-freezing and 7% of glycerol concentration. These results lead to commercial AI with frozen-thawed stallion semen.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.61-65
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• 2011

The objective of this study was to examine the effect of various discontinuous Percoll washing conditions on motile sperm recovery rate and motion kinematics. Frozen semen samples from 3 bulls (0.5 ml plastic straws, 6% glycerol in egg yolk-Tris-glycerol extender) were thawed in $37^{\circ}C$ water bath for 1 min. After thawing, the mixed semen samples were randomly allocated to 12 treatment groups. Briefly, the spermatozoa were centrifuged for three different time lengths (10, 20, and 30 min) at two gravities ($300{\times}g$ and $700{\times}g$) through two concentrations of discontinuous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll and 2 ml 90%: 2 ml 45% Percoll to remove extender, debris, and dead spermatozoa. Motile sperm recovery rate and motion kinematics were evaluated by computer assisted sperm analyzer using Makler counting chamber. Sperm motility (%) and motile sperm recovery rate showed similar pattern in all treatment groups. However, sperm motility (%) and motile sperm recovery rate were highest at $700{\times}g$ for 30 min through a discontionous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll. There were no significant differences in motion kinematics after various Percoll washings. These results suggest that force of centrifugation, centrifugation time, and Percoll volume significantly affect motile sperm recovery rate.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.35 no.5
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• pp.62-71
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• 2003

This study was carried out to develop the bio-chemical pulping method to enhance the energy saving and decrease the capital cost through the soda pulping under atmospheric pressure ($100^{\circ}C$). Nonwood substrates, rice straw, were pretreated by white-rot fungi, Pleurotus cornucopiae. Several basic pieces of data that can be applied in soda pulping were acquired. The results of this study were as follows. Under the conditions without any nutrients or with glucose, N and glucose + N, the weight losses of rice straws inoculated by Pleurotus cornucopiae were 12.1∼32.6 %, 12.0∼26.3 %, 13.0∼25.4 % and 15.3∼24.7 % for 5, 10, 15, 20, 25 and 30 days incubation periods respectively. The more the fungal incubation was extended, the more the weight losses were gained. The yield of untreated rice straw was 54.8 % after pulping. When any nutrients was not added or glucose, N and glucose ＋ N were added for the pretreatment, the total yields were ranged to 57.3∼42.9 %, 51.0∼43.3 %, 51.7∼43.9 % and 52.1∼46.1 % for 5 different incubation periods respectively. The yields were gradually decreased based on the extending of the incubation periods. The physical properties of the rice straw soda pulp without fungal treatment, the density, breaking length, burst index, tear index and folding endurance were 0.24g/㎤, 2.32 Km, 0.91 kPaㆍ$m^2$/g, 46.7 mNㆍ$m^2$/g and 21 times, respectively. In the case of pretreatment without any nutrients or with glucose, N and glucose + N as nutrients, the density was 0.24g/㎤, the breaking length was 3.30∼6.46 Km, the burst index was 1.36∼3.01 kPaㆍ$m^2$/g, the tear index was 33.0∼57.0 mNㆍ$m^2$/g and the folding endurance was 14∼381 times at most incubating periods, when pulping was done. The physical properties were increased as the incubation duration was extended. Especially, when N and glucose ＋ N were added, the physical properties showed superior results during each incubation period.

• The Korean Journal of Pesticide Science
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• v.20 no.4
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• pp.341-348
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• 2016

Residual patterns of fungicides fludioxonil and metconazole applied 2 or 3 times on wheat were investigated with consideration for their distribution rates in each compartment of wheat. Wheat samples collected at harvesting day were divided into three compartments such as grain, hull and straw, and the pesticide residue analysis was individually carried out to such compartments of wheat. The analytical methods of fungicide residues in wheat were acceptable, with recoveries of 84.3 to 113.3% and correlation coefficients of 0.1 to 4.7%. Due to the systemic characteristics, the residual amounts of metconazole in the grain part of wheat were greater as 0.13~0.17 mg/kg than those amounts (0.01-0.03 mg/kg) for fludioxonil. To absolute residue amounts of the fungicides in one wheat plant, their distribution rates in each compartment of wheat were the highest in straws (68.5-70.7%), followed by hulls (29.0-31.0%) and grains (0.2-0.8%).

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.2
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• pp.230-240
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• 2016

Information on the effects of different yeast species on ruminal fermentation is limited. This experiment was conducted in a $3{\times}4$ factorial arrangement to explore and compare the effects of addition of three different live yeast species (Candida utilis 1314, Saccharomyces cerevisiae 1355, and Candida tropicalis 1254) at four doses (0, $0.25{\times}10^7$, $0.50{\times}10^7$, and $0.75{\times}10^7$ colony-forming unit [cfu]) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover, and rice straw by mixed rumen microorganisms in dairy cows. The maximum gas production (Vf), dry matter disappearance (IVDMD), neutral detergent fiber disappearance (IVNDFD), and methane production in C. utilis group were less (p<0.01) than other two live yeast supplemented groups. The inclusion of S. cerevisiae reduced (p<0.01) the concentrations of ammonia nitrogen ($NH_3$-N), isobutyrate, and isovalerate compared to the other two yeast groups. C. tropicalis addition generally enhanced (p<0.05) IVDMD and IVNDFD. The $NH_3$-N concentration and $CH_4$ production were increased (p<0.05) by the addition of S. cerevisiae and C. tropicalis compared with the control. Supplementation of three yeast species decreased (p<0.05) or numerically decreased the ratio of acetate to propionate. The current results indicate that C. tropicalis is more preferred as yeast culture supplements, and its optimal dose should be $0.25{\times}10^7$ cfu/500 mg substrates in vitro.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.35-40
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• 2013

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Holstein Donor Cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_2{\alpha}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II(first ovulation did not occur until ${\geq}40$ days after superovulation, i.e. delayed first ovulation 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and $10{\leq}$ of corpora lutea(CL) was $7.8{\pm}1.8$ and $12.7{\pm}2.7$, respectively. The number of transferable embryos differed between < 10 and $10{\leq}$ CL was $5.4{\pm}1.3$ and $8.1{\pm}3.4$, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. Diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.23-28
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• 2003

The present study was undertaken to investigate the effects of PVP concentration and exposure temperature to vitrification solution on the post-thaw survival, in vitro maturation and development of immature bovine oocytes (germinal vesicle stage). The vitrification solution (VS) consisted of 40% ethylene glycol (EG)+0.5 M sucrose (S)+10% FBS. PVP was added to VS: 0%, 5% or 10%. The cumulus-oocyte complexes (COCs) were diluted in VS as one step, after 2 min the COCs were loaded in straw and vitrified by direct immersion into liquid nitrogen. For thawing, the straws were plunged into $30^{\circ}C$ water bath for 10s. After thawing, the oocytes were diluted in 0.5 M (in DPBS with 10% FBS) sucrose solution for 5 min. The survival rate (FDA-test and trypan blue) of immature bovine oocytes was measured. The survival rate was higher in 5% PVP (91.5%) than in 0% (64.2%) or in 10% PVP (79.7%). The proportion of metaphase II formation was 69.35% in control (no vitrified COCs), 9.3% in 40% EG+0.5 M S+0% PVP and 21.05% in 40% EG+0.5 M S+5% PVP (p<0.05). The effect of room temperature ($25^{\circ}C$ for 10 min) and cold temperature ($4^{\circ}C$ for 10 min) on COCs were determined in this study. After IVF, the cleavage and blastocysts rate of oocytes exposed to room temperature and cold temperature in VS+5% PVP was significantly different (2 cell: 63.20% vs 37.97%, blastocysts: 18.40% vs 2.53%). The cleavage rates of frozen-thawed oocytes were 20.53% with PVP and 22.13% without PVP (p>0.05). Two out of 151 oocytes (1.32%) developed to blastocyst stage after frozen-thawed with 5% PVP (p>0.05). Development of oocytes after frozen-thawing to the 2 cell were not significantly affected with or without PVP following IVF. However, the vitrification of immature bovine oocytes with PVP maintained the ability to develop to the blastocyst stage after IVM-IVF and IVC, while no blastocysts were obtained from oocytes vitrified without PVP. These results suggested that PVP has a protective role for vitrification of immature bovine oocytes as far as survival is concerned, however, the protection was not sufficient enough to support blastocyst formation.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.66-74
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• 2012

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

• Magazine of the Korean Society of Agricultural Engineers
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• v.17 no.3
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• pp.3795-3814
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• 1975

The purpoes of this thesis is to study the effect of the variation of the beginning date of the suspension of irrigation and the length of intermittent suspension period of irrigation in the paddy field on the growth and yield of rice, so as the provide a critical limit of saving irrigation water and an irrigation method to prevent drough damage in rice cultivation. In this experiment, the rice variety adopted was NONGRIM No.29. There were seven main test plot, each test plot having a different beginning date of the suspension of irrigation. A main test plot was susdivided into five small test plots, each having a different length of the suspension period of irrigation. The results obtained in this experiment are summarized as follows: 1. The number of tillers is controlled by the treatment of the suspension of irrigation, its beginning date being early of late. The reductive effects of beginning dates of suspension upon the number of tillers. investigated on July 30, are about 84% for the treatment suspended on July 8, 87% on July 12, and 92% on July 19, respectively, in comparison with the standard plot. However. the suspenion treatments after the foregoing dates does not affect the control of their numbers. On the other hand, the lengths of intermittent suspension periods influence highly on the number of tillers to be restrained considerably, the decrease ratio ranging from 91% to 80% Both treatments on dates and periods are so intermingled that the restraining effects of suspension periods become greater as the dates of its beginning are earlier. 2. The elongation of plant hieghts also restrained considerably by the longer periods and earlier dates of suspension treatments of irrigation. Especially, the effects of the lengths of suspension periods become more serious. 3. Heading dates are delayed by two to five days through the suspension treatments of irrigation. However, the heading stage ends almost on the same day without relation to the differences between the irrigation suspension treatments. In the test plot where the suspension date of irrigation comes after the young panicle forming stage, the heading stage ends one or two days later than in the standard test plot. 4. Both culm lengths and panicle lengths show significant differences in their values, i.e., their lengths are shorter, as the begining dates of irrigation suspension are earlier and the suspension periods are longer. 5. The earlier the beginning date and the longer the period of irrigation suspension, the less is the number of panicles per hill in comparison with the standard plot. 6. The earlier the beginning date and the longer the period of irrigation suspension, the higher is the significant difference in the number of kernels per panicle in comparison with that in the standard test plot, i.e., the less is the number of kernels per panicle. 7. The earlier the beginning date and the longer the period of irrigation suspension, the lighter are the weights of rough rice and straws per hill in comparison with those in the standard plot.