• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1562-1570
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• 2014

The effects of Lactobacillus mucosae (L. mucosae), a potential direct fed microbial previously isolated from the rumen of Korean native goat, on the rumen fermentation profile of brewers grain were evaluated. Fermentation was conducted in serum bottles each containing 1% dry matter (DM) of the test substrate and either no L. mucosae (control), 1% 24 h broth culture of L. mucosae (T1), or 1% inoculation with the cell-free culture supernatant (T2). Each serum bottle was filled anaerobically with 100 mL of buffered rumen fluid and sealed prior to incubation for 0, 6, 12, 24, and 48 h from which fermentation parameters were monitored and the microbial diversity was evaluated. The results revealed that T1 had higher total gas production (65.00 mL) than the control (61.33 mL) and T2 (62.00 mL) (p<0.05) at 48 h. Consequently, T1 had significantly lower pH values (p<0.05) than the other groups at 48 h. Ammonia nitrogen ($NH_3$-N), individual and total volatile fatty acids (VFA) concentration and acetate:propionate ratio were higher in T1 and T2 than the control, but T1 and T2 were comparable for these parameters. Total methane ($CH_4$) production and carbon dioxide ($CO_2$) were highest in T1. The percent DM and organic matter digestibilities were comparable between all groups at all times of incubation. The total bacterial population was significantly higher in T1 (p<0.05) at 24 h, but then decreased to levels comparable to the control and T2 at 48 h. The denaturing gradient gel electrophoresis profile of the total bacterial 16s rRNA showed higher similarity between T1 and T2 at 24 h and between the control and T1 at 48 h. Overall, these results suggest that addition of L. mucosae and cell-free supernatant during the in vitro fermentation of dried brewers grain increases the VFA production, but has no effect on digestibility. The addition of L. mucosae can also increase the total bacterial population, but has no significant effect on the total microbial diversity. However, inoculation of the bacterium may increase $CH_4$ and $CO_2$ in vitro.

• Journal of the Korea Organic Resources Recycling Association
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• v.23 no.3
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• pp.85-92
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• 2015

For concerning the climate change issues, the carbon sequestration and importance of soil organic matter are receiving high attention. To evaluate carbon sequestration in soil is important to determine the soil organic carbon (SOC) fractions such as WESOC (Water extractable soil organic carbon), and $CO_2$ emission by soil microbial respiration. However, the analyses for those contents are time-consuming procedure. There were studied the feasibility of MIRS (Mid-Infrared Spectroscopy), which has short analysis time for determining the WESOC and an incubated carbon in this study. Oven-dried soils at $100^{\circ}C$ and $350^{\circ}C$ were scanned with MIRS and compared with the chemically analyzed WESOC and cumulative carbon dioxide generated during 30, 60, 90, and 120 days of incubation periods, respectively. It was observed that an optimized determination coefficient was 0.6937 between WESOC and untreated soil processed by spectrum vector normalization (SNV) and 0.8933 between cumulative $CO_2$ from 30 days incubation and soil dried at $350^{\circ}C$ after subtracting air-dried soil processed by 1st derivatives. Therefore, it was shown that Quantification of soil organic carbon fractions was possibility to be analyzed by using MIRS.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.133-139
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• 1997

Large scale production of cloned embryos requires the technology of multiple generational nuclear transfer(NT) by using NT embryos itself as the subsequent donor nuclei. In this work we investigated comparatively the effects of enucleated oocytes treated with ionomycin and 6-DMAP on the electrofusion rate and in vitro developmental potential in the first and second NT embryos. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FCS) at 47 hours after hCG injection. The recipient cytoplasms were obtained by removing the nucleus and the first polar body from the oocytes collected at 15 hours after hCG injection. The enucleated oocytes were pre-activated by 5 min incubation in 5$\mu$M ionomycin and 2 hours incubation in 2 mM 6-DMAP at 19~20 hours post-hCG before microinjection. In the first and second generation NT, the unsynchronized 16-cell stage embryos were used as nuclear donor. The separated donor blastomeres were injected into the enucleated activated recipient oocytes by micromanipulation and were electrofused by electrical stimulation of single pulse for 60 $\mu$sec at 1.25kV/cm in $Ca^2$+, $Mg^2$+ - free 0.28 M mannitol solution. In the non-preactivation group, the electrofusion and electrical stimulation was given 3 pulses for 60 $\mu$sec at 1.25 kV/cm in 100$\mu$M $Ca^2$+, $Mg^2$+ 0.28 M mannitol solution. The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in TCM-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The results obtained were summarized as follows: 1. In the first generational NT embryos, the electrofusion rate of preactivated and non-activated oocytes(80.4 and 87.8%) was not significantly different, but in the second generational NT embryos, the electrofusion rate was significantly(P<0.05) higher in the non-activated oocytes(85.7%) than in the preactivated oocytes(70.1%). 2) In the first and second generational NT embryos, the developmental potential to biastocyst stage was significantly(P<0.05) higher in the preactivated oocytes(39.3 and35.7%) than in the non-preactivated oocytes(16.0 and 13.3%). No significant difference in the developmental potential was shown between the first and second generational NT embryos derived from the preactivated oocytes. In conclusion, it may be efficient to use the oocytes preactivated with ionomycin and 6-DMAP for the multiple production of cloned embryos by recycling nuclear transfer.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.330-335
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• 1999

Pentachlorophenol(PCP), which is very persistent in soil and water environment, was tried to detoxify with oxidoreductive catalysts(peroxidase, laccase, tyrosinase and birnessite). To find out detoxification of PCP, the transformation of PCP through oxidative coupling was investigated in the presence of various oxidoreductive catalysts. PCP incubated with peroxidase was significantly transformed, however, in case of tyrosinase, the transformation was negligible. Using peroxidase, the optimal reaction condition was pH 5.6 and $16^{\circ}C$. The transformation of PCP was very fast in initiation step until 30 min but, that was not observed after 180 min. The transformation of PCP was increased by increasing peroacidase amount. When the effect of humic monomer was investigated as co-substrate on the transformation of PCP, the transformation of PCP was mostly decreased in the incubation with peroxidase, laccase, and birnessite. The transformation of PCP, however, was slightly increased by the incubation with tyrosinase in the presence of humic monomers as co-substrate, except catechol. On the basis of the results obtained, it may be suggested that PCP is able to be effectively detoxified through oxidative coupling mediated with oxidoreductive catalysts.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.187-193
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• 1998

The effects of medium, incubation temperature, incubation period, pH of medium and $CO_2$ ondition during mycelial growth were investigated to study the factors associated with the formation of oidia in Flammulina velutipes. Oidia formation was increased when mycelial growth was poor, while oidia formation was inhibited in optimum condition of mycelial growth. Mating type of oidia was investigated to examine the effect of oidia formation on dikaryotic strain. Di-mon matings between oidia strains and original dikaryotic strain were carried out. Monokaryotic strains derived from oidia showed only one genotype. Seventy percent among Dimon mating strains showed slow mycelial growth and low yield of fruit-body, but others showed similar or high mycelial growth and yields in comparision with original dikaryon strain. One strain from di-mon mating demonstrated some differences in isozyme band pattern.

• Korean Journal of Food Science and Technology
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• v.34 no.1
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• pp.73-78
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• 2002

Bacillus polyfermenticus SCD, which is commonly called as Bisroot strain, is being used for functional foods through the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine in both humans and animals. The cells of B. polyfermenticus SCD were treated for 24 h in artifical bile after incubation for 2 h in artificial gastric juice and final number of the strain was reached to around $3.3{times}10^7\;CFU/mL$. In test of API ZYM kit, ${\beta}-glucuronidase$ or ${\beta}-glucosidase$ was not produced by B. polyfermenticus SCD. B. polyfermenticus SCD was resistant to antibiotics, such as nisin, streptomycin, tetracycline, and rifamycin. B. polyfermenticus SCD was also affected by alcohol concentration up to 4%, but more than 8%, their growth was not affected significantly. Finally, B. polyfermenticus SCD was shown to inhibit the growth of Listeria monocytogenes ATCC 19111 completely within 24 h of incubation, which indicated its bactericidal nature.

• Korean Journal of Weed Science
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• v.30 no.4
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• pp.421-428
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• 2010

A foliage contact herbicidal substance was separated from ethyl ether fraction in n-hexane extract of Saussurea lappa roots and identified as dehydrocostus lactone [(3aS,6aR,9aR,9bS)-3,6,9-trimethylidene-3a,4,5,6a,7,8,9a,9b-octahydroazuleno[5,4-d]furan-2-one](DHCL). When DHCL at 4,000 ppm was foliage-applied to two grasses and two broadleaf plants, greater than 85% necrotic injury was obtained from large crabgrass, maize and soybean, whereas only about 40% necrotic injury appeared in black nightshade, indicating that DHCL has no gross morphological selectivity, but shows difference in contact response among the plant species tested. Conductivity in incubation medium of the leaf disks treated with DHCL increased as the incubation time continued. Relatively low contact injury in black nightshade as compared with the other three plant species tested was attributed to decrease in absorption of DHCL due to relatively high amount of cuticle. DHCL did not require light in the herbicidal action and there were no inhibitory effects on seed germination and cell elongation. Acetyl-CoA carboxylase activity was inhibited by 30% and 58% at $100\;{\mu}M$ and $1000\;{\mu}M$ DHCL, respectively. These results suggested that the herbicidal action of DHCL was related with inhibition of fatty acid synthesis which in turn caused to weaken cell membrane integrity.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.352-357
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• 2008

We evaluated cultivation methods to obtain pure cultures of previously uncultivated bacteria from soil. Soil bacteria (suspensions) were inoculated onto various oligotrophic media with one of the following additives: 1) soil extract; 2) anthraquinone disulfonate (humic acid analogue); 3) acyl homoserine lactones (quorum-signaling compounds); 4) catalase (for the protection of bacteria from exogenous peroxides). After the relatively long period (60 days) of incubation with elevated concentrations of $CO_2$ (5%, v/v), the media containing catalase showed the highest colony count. We purified 147 randomly selected colonies from the media and the isolates were subjected to the phylogenetic analyses of their 16S rRNA gene sequences. Phylogenetic analysis revealed that approximately 30% of the isolates might belong to novel species or novel family, suggesting that the media and incubation conditions used could be useful for the cultivation of as-yet-uncultured bacteria. Especially, bacteria belonging to the phylum Acidobacteria, ubiquitous bacterial taxon known as an uncultured bacterial group (at least difficult to culture from environmental samples), were successfully cultured in this study.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.4
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• pp.276-285
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• 2022

This study evaluated the effect of lactic acid bacteria (LAB, a mixture of Enterococcus faecium and Lactobacillus plantarum) supplementation, the storage temperature, and storage period on the fermentation characteristics and in vitro ruminal digestibility of a total mixed ration (TMR). The TMR was prepared into two groups, namely, CON (control TMR without the LAB) and ML (supplementing a mixture of E. faecium and L. plantarum in the ratio of 1% and 2% (v/w), respectively). Both groups were divided and stored at 4℃ or 25℃ for 3, 7, and 14 d fermentation periods. Supplementing LAB to the TMR did not affect the chemical composition of TMR except for the lactate and acetate concentration. Storage temperatures affected (p<0.05) the chemical composition of the TMR, including pH, lactate, and acetate contents. The chemical composition of TMR was also affected (p<0.05) by the storage period. During in vitro rumen fermentation study, the ML treatment showed lower (p<0.05) dry matter digestibility at 24 h incubation with a higher pH compared to the CON. There was no difference in the in vitro dry matter digestibility (IVDMD) of TMR between the CON and ML treatment however, at 24 h, ML treatment showed lower (p<0.05) IVDMD with a higher pH compared to the CON. The effects of storage temperature and period on IVDMD were not apparent at 24 h incubation. In an in vivo study using Holstein steers, supplementing LAB to the basal TMR for 60 d did not differ in the final body weight and average daily gain. Likewise, the fecal microbiota did not differ between CON and ML. However, the TMR used for the present study did include a commercial yeast in CON, whereas ML did not; therefore, results were, to some extent, compromised in examining the effect of LAB. In conclusion, storage temperature and period significantly affected the TMR quality, increasing acetate and lactate concentration. However, the actual effects of LAB supplementation were equivocal.

• Korean Journal of Microbiology
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• v.40 no.3
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• pp.244-247
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• 2004

To investigate the effects of elevated $CO_2$ on the denitrifying bacterial community structure in a wetland soil, dynamics of bacterial community structure was explored in an artificial wetland ecosystem with one of three plant species (T. latifolia, S. lacustris, and 1. effusus) under two levels of $CO_2$(370 ppm or 740 ppm) after 110day incubation. For the analysis of bacterial community structure, functional genes such as nitrite reductase genes (nirS) were PCR-amplified followed by cloning of PCR products and screening by restriction fragment length polymorphism (RFLP). nirS gene fragments were amplified in all analyzed soil samples. Species richness estimated by the number of distinct phylotypes were 83 and 95 in the ambient $CO_2$ treatment and the elevated treatment, respectively. Two phylotypes (type 1 and type 2) were dominant in both of the treatments. Elevated $CO_2$ treatment increased species richness of denitrifying as well as changed a large proportion of denitrifier phylotypes compared to those of the ambient treatment. Overall, the data in this study suggested that the denitrifying communities in the wetland soil are diverse and that the richness of denitrifying bacterial community might be affected by elevated $CO_2$ treatment.