Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Cho, Chang-Yeon;Suh, Sangwon;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
Korean Journal of Poultry Science
/
v.41
no.4
/
pp.261-270
/
2014
This study was conducted to establish the method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of simple freeze-thaw treatment on viability of PGCs in chickens and to the optimal protocol for PGCs freezing. PGCs obtained from the germinal gonade of an early embryos of 5.5~6 day (stage 28) of Isa Brown, Korean Ogye (KO), White Leghorn and Commercial breeds, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15%, and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to simple freezing, with different concentrations of the cryoprotectant solution, were examined. After simple freezing, the viability of PGCs after freeze-thawing was significantly higher for Commercial breeds ($88.7{\pm}2.4%$) than KO ($85.1{\pm}0.4%$), Isa Brown ($84.6{\pm}0.2%$) and White Leghorn ($85.9{\pm}0.1%$) (p<0.05) using 10% EG cryoprotectant. Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a database.
Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Byun, Mi Jeong;Kim, Young Sin;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
Korean Journal of Poultry Science
/
v.41
no.4
/
pp.249-259
/
2014
Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.
Kim, Kwang-Sik;Kim, Kil-Yong;Son, Bo-Gyun;Lee, Young-Hwan;Kim, Yong-Woong;Seong, Ki-Young
Korean Journal of Soil Science and Fertilizer
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v.25
no.2
/
pp.181-188
/
1992
This study was carried out to evaluate the fate of inoculant Bradyrhizobium japonicum and the inoculation effect on soybean in complex soil environment. To moniter Rhizobium strains from the root, streptomycine and streptomycine and nalidixic acid resistant marker strains were prepared by spontaneous mutagenesis. The characteristics and properties of antibiotic marked strains were not altered by the mutagenesis. The comparison of properties of wild type and antibiotic resistant Bradyrhizobium strains are summarized as follow : 1) The strains of USDA110K-$STR^r$, USDA110N-$STR^r$ and R318-$STR^r$ showed weak tolerance to pH 9.0. The utilization of carbon sources by fast growing group was different from that of slow growing group. The marked strains of R214-$STR^rNAL^r$, USDA110K-$STR^r$ and USDA110N-$STR^r$ was doubtful in utilization of sorbitol and R138-$STR^rNAL^r$ was doubtful in utilization of xylose as a carbon source. 2) By examining the agglutination reaction of serogroups, the strains used were identified as different ones. There were no differences between wild type and marked strains in agglutination titer values. 3) The plasmid size of fast group was slightly greater than that of slow group. However, there was no differences in plasmid size between the wild type and antibiotic resistant strains. This result indicates that the antibiotic resistance was not encoded in plasmid. 4) The recovery of the inoculated strains was up to 12.5 % in soybean cultivated soil and was up to 25 % in soybean uncultivated soil. 5) When the wild type or marked strains were inoculated. there was no significant effect on soybean plant, whereas the inoculation effect was pronounced in soybean uncultivated soil. The inoculation effect seemed to be more pronounced in wild type strains than antibiotic resistant strains. however, the difference was not significant.
These study were conducted to determine the effects of a whole or steam-flaked corn based diet on rumen microbial fermentation in vitro and ruminal metabolism in the Korean Native Goat(KNG) in vivo. The experiments consisted of two dietary treatments: control, steam-flaked corn(SFC) based diet(80%) + rice straw mixed(20%)(SFCR); 100% whole corn based diet(WC). The first experiment was conducted to investigate the effect of whole corn on ruminal metabolism in vitro for 0 to 48 h. pH values were optimally maintained during incubation time, and were not significantly different between treatments. Gas production of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration tended to increase for WC, but not significantly different between treatments. The mean value of total volatile fatty acid concentration of WC was significantly lower than SFCR(p<0.01), but SFCR and WC linearly increased as the time of incubation approached 48 h. Mean value of acetate concentration of SFCR was significantly higher than WC(p<0.01). Propionate concentration of WC for the total incubation time was significantly higher than SFCR(p<0.01). The digestibility of dry matter was not significantly different between treatments, but SFCR was somewhat higher than WC. The second experiment was conducted to effect of whole shelled corn based diet on rumen metabolism in KNG. pH values tended to decrease through all treatments. There was not a significantly difference between treatments. Microbial protein yield of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration of WC was significantly (p<0.01) higher than SFCR. Total VFA and propionate concentration of WC was significantly higher than SFCR(p<0.01), but acetate concentrate of WC was not significantly higher than SFCR. The mean value of total lactate concentration was significantly(p<0.01) different but the value of SFCR and WC were lower than the average concentration of acidosis. In sacco DM disappearance rate of SFC was significantly(p<0.01) higher than WC.
This study was conducted to investigate the effects of dietary full-fat soybeans and linseed as fat sources on in vitro ruminal disappearances of dry matter and unsaturated fatty acids and fatty acids profile. The full-fat soybeans and linseed were high in linoleic acid (C18:2n-6) and $\alpha$-linolenic acid (C18:3n-3), respectively. The incubation times were 0, 3, 6, 12, 24, 48 and 72 h. After each time of incubation, medium digesta was lyophilized for analyzing its DM and fatty acids contents. DM disappearance was significantly higher in linseed treatment compared to full-fat soybeans treatment on 6 h (p<0.01), 12 h (p<0.05) and 24 h (p<0.01), but cumulative gas production was not significantly different between both treatments. Stearic acid (C18:0) content in medium digesta was increased in both soybeans and linseed as a result of complete biohydrogenation with increased incubation time and C18:0 and C18:1 contents of full-fat soybeans were significantly higher than those of linseed (p<0.05). The content of C18:2 and C18:3 in digesta of each treatment were decreased by biohydrogenation as incubation time was increased. The content of C18:2 in full-fat soybeans was significantly higher than that of linseed (p<0.05) while the content of C18:3 in linseed was significantly higher than that of full-fat soybeans (p<0.001). Net C18:0 production was significantly higher in full-fat soybeans (332.24%) than linseed (133.16%) on 72 h. Disappearance of C18:1 was significantly lower in full-fat soybeans than linseed (p<0.05), especially full-fat soybeans showed negative (-) values on 3, 6, 12 and 24 h. The disappearance of C18:3 was significantly higher in linseed than full-fat soybeans (p<0.05). The disappearance of C18-unsaturated fatty acid was significantly higher in linseed than full-fat soybeans. In conclusion, polyunsaturated fatty acid (PUFA) in both full-fat soybeans and linseed were extensively biohydrogenated. In addition, biohydrogenation of PUFA was more completed to C18:0 in full-fat soybeans than linseed, reflecting dietary PUFA composition.
To comply with stricter regulations provoked by increasing odor nuisance, it is imperative to practice effective odor control for sustainable livestock production. This study was conducted to assess odor and odorous compounds emitted from liquid animal manure with different treatment methods such as Fresh Manure(without treatment, FM), Anaerobic Digestion(AD) and Thermophilic Aerobic Digestion(TAD) and their application to soil. Air samples were collected at the headspace of liquid manure, upland and paddy soil, and analyzed for odor intensity and offensiveness using an olfactometry; odor concentration index using odor analyser; nitrogen-containing compound such as ammonia(NH3) using fluorescence method; and sulfur containing compounds such as hydrogen sulfide(H2S), methyl mercaptan(MeSH), dimethyl sulfide(DMS) and dimethyl disulfide(DMDS) using gas chromatography-pulsed flame photometric detector, respectively. Odor intensity, offensiveness and concentration index from TAD liquid manure was statistically lower than those from FM and AD(p<0.01). Mean concentrations of H2S, MeSH, DMS, DMDS and NH3 were 65.93ppb, 18.55ppb, 5.26ppb, 0.33ppb and 10.57ppm for liquid manure with AD; and 5.15ppb, 0.97ppb, 0.80ppb, 0.56ppb and 1.34ppm for liquid manure with TAD, respectively. More than 60% of malodorous compounds related to nitrogen and sulfur were removed by heterotrophic microorganisms during TAD treatment. When liquid manure was applied onto upland and paddy soil, NH3 removal efficiencies ranged from 51 to 94% and 22 to 91% for AD and TAD liquid manure, respectively. The above results show that liquid manure with TAD is superior to AD and FM with respect to the odor reduction and odor problem caused by land applied liquid manure is directly related to the degree of odor generated by the manure treatment method.
Proceedings of the Korean Society for Applied Microbiology Conference
/
2005.06a
/
pp.154-164
/
2005
Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.
Investigation of the $CO_2$ exchange between biosphere and atmosphere at regional, continental, and global scales can be directed to combining remote sensing with carbon cycle process to estimate vegetation productivity. NASA Earth Observing System (EOS) currently produces a regular global estimate of gross primary productivity (GPP) and annual net primary productivity (NPP) of the entire terrestrial earth surface at 1 km spatial resolution. While the MODIS GPP algorithm uses meteorological data provided by the NASA Data Assimilation Office (DAO), the sub-pixel heterogeneity or complex terrain are generally reflected due to coarse spatial resolutions of the DAO data (a resolution of $1{\circ}\;{\times}\;1.25{\circ}$). In this study, we estimated inputs retrieved from MODIS products of the AQUA and TERRA satellites with 5 km spatial resolution for the purpose of finer GPP and/or NPP determinations. The derivatives included temperature, VPD, and solar radiation. Seven AmeriFlux data located in the Corn Belt region were obtained to use for evaluation of the input data from MODIS. MODIS-derived air temperature values showed a good agreement with ground-based observations. The mean error (ME) and coefficient of correlation (R) ranged from $-0.9^{\circ}C$ to $+5.2^{\circ}C$ and from 0.83 to 0.98, respectively. VPD somewhat coarsely agreed with tower observations (ME = -183.8 Pa ~ +382.1 Pa; R = 0.51 ~ 0.92). While MODIS-derived shortwave radiation showed a good correlation with observations, it was slightly overestimated (ME = -0.4 MJ $day^{-1}$ ~ +7.9 MJ $day^{-1}$; R = 0.67 ~ 0.97). Our results indicate that the use of inputs derived MODIS atmosphere and land products can provide a useful tool for estimating crop GPP.
The study was conducted to investigate the effects of climate on indoor environment of a swine house with natural. This study was tested in the beef swine stall at Young-in, Kyung-ki do. The test was experimented for the effect of interior environment by the outdoor environment and the interior-pan. The results are as follows. 1. In test 1 ($T_{out}$ : $25.7^{\circ}C$, without fan), an indoor air flow pattern was showed that entered from sidewall winch-curtain to went out of a indoor by the ridge winch-curtain. And the velocity of a section of the center was measured two times as large as the velocity of the floor. It is the acceleration of the velocity by thermal buoyancy. And, the entered air was rapidly dissipated by flow energy. So that in the swain livestock with sidewall winch-curtain is effected by thermal buoyancy. And the air temperature of the indoor was distributed more higher as compared with the outdoor temperature. This result is caused by the sensible heat from swine and the ventilation is restricted. 2. In test 2 (($T_{out}$ : $25.7^{\circ}C$, with fan), the velocity of a section of the center was measured more higher as compared with the test 1. And the variance of air velocity was distributed higher as compared with the test 1. This result is showed dead region of air flow with a fan operation. And, the variance of gas density was distributed lower as compared with the test 1.
The objective of this study was to investigate the effect of chronic unpredictable stress on the reproductive function and ovarian luteinizing hormone receptor (LHR) expression. 9-week-old C57BL/6 female mice were randomly divided into two groups: control group and stressed group. Mice have been stressed twice a day for 35 days with 12 different stressors which were randomly selected. The results demonstrate that there is significant increase in the anxiety-related behaviors (P < 0.05), decrease body weight gain rate (P < 0.01) and decrease in the average of litter size in stressed mice compared with control group (P < 0.01). Furthermore, the rate of primary, secondary and early antral follicles in stressed mice significantly decreased (P < 0.05), whereas that of atretic follicles significantly increased compared with control mice (P < 0.01). The immunohistochemical analysis revealed that reduced LHR expression in granulosa cells of follicle and luteal cells of corpus luteum in response to chronic unpredictable stress. The western blot analysis revealed significantly decrease in LHR expression in the stressed mice ovaries compared with the control (P < 0.05). These results suggest that ovarian LHR expression affected by chronic unpredictable stress and the modulated ovarian LHR is responsible for ovarian follicular maldevelopment and reproductive dysfunction.
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