The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).
Park, Mi Na;Jang, Hyun-Jun;Keum, Dae Ho;Choi, Jin Ae;Yoo, Jae Gyu;Byun, Sung June;Park, Jong Ju;Ji, Ju Young;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong
Korean Journal of Poultry Science
/
v.40
no.4
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pp.299-304
/
2013
Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.
This study was conducted to examine in vitro development of porcine embryos constructed by the microinjection of cultured fetal fibroblast cells into porcine oocytes matured in vitro. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at 38.5$^{\circ}C$ for 6 to 8 days in 5% $CO_2$ and air. In experiment 1, fusion rates of nuclear transfer embryos did not differ for fetal fibroblast cells incubated in 5% FBS + NCSU-23 or 5% FBS + TL Heaps medium, nor did fusion rates of donor cells differ between 1-8 hr incubation durations. Fusion rates for the four treatment subclasses ranged from 72.1% to 78.0%. In experiment 2, Pre-synchronization in medium containing 0.1 $\mu\textrm{g}$/m Hoechst 33342 an increase from 0 and 8 versus 15 h culture an increased percentage of porcine fibroblast cells in G2/M at the end of the synchronization period (12.4%, 17.5% and 47.6%). Neither an increase in the concentration of H 33342 (0.2-1.6 $\mu\textrm{g}$/$m\ell$) nor a longer exposure time (12h, 18h and 24h) increased the proportion of porcine G2/M fibroblasts. In experiment 3, fusion rates did not differ significantly far nuclear transfer embryos constructed using donor cells cultured in 5% FBS + NCSU-23 medium for 1-2, 6-8 or 12-14 days (60.0%, 73.3% and 62.5%), respectively. The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1-2 days was 44.0%, significantly less than 56.7% and 50.0%. for 6-8 or 12-14 days duration of culture, respectively. In experiment 4, the proportions of nuclear transfer embryos that developed to the $\geq$2 cell and to the blastocyst stage were not affected significantly by culture medium (5% FBS + NCSU-23 or 5% FBS + TL-Heaps) or by $O_2$ concentration of the culture (5% vs 10%). Rates of development to the $\geq$2 cell stage ranged from 65.9% to 70.1%, and development rates to the blastocyst stage ranged from 9.8% to 12.5% for the four treatment subclasses. Developmental rate was highest for embryos cultured in 5% FBS + NCSU-23 under a gas atmosphere of 5% $O_2$ in air.
The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.
Oh, Seung Min;Kim, Ji Yun;Lee, Bae Hun;Peng, Jinglun;Chemere, Befekadu;Nejad, Jalil Ghassemi;Sung, Kyung Il;Kim, Byong Wan
Journal of The Korean Society of Grassland and Forage Science
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v.36
no.2
/
pp.124-128
/
2016
The objective of this research was to determine the effects of harvesting frequency and fertilization levels on botanical composition, dry matter yield, and forage feed compositions of Alpine grassland at 800 m altitude. This research lasted for three years at National Alpine Agricultural Research Institute in Pyeongchang with two harvesting frequency schedules (two and three times annually) and two levels of fertilizer application (conventional level of fertilizer at 280-200-240 kg/ha and a lower level of fertilizer at 200-200-200 kg/ha for N, $P_2O_5$, and $K_2O$). Mixture combinations with seeding rate (kg/ha) were as follows: Orchardgrass 18, Tall fescue 9, Timothy 8, Kentucky bluegrass 3, and Ladino Clover 2. The gramineae ratio ranged from 93.2 to 95.3%. Therefore, gramineae forage was considered as the dominant plant in this experiment. No significant (p>0.05) difference was observed in forage dry matter yield between the two harvesting frequency treatments (two times at 9.8 ton/ha and three times at 8.6 ton/ha). However, forage dry matter yield in the two times of harvesting frequency tended to be greater than that in the three times of harvesting frequency. Significantly (p<0.05) higher forage dry matter yield in the standard fertilization level group than the lower fertilization level group (9.8 ton/ha vs. 8.7 ton/ha) was observed. However, there was no significant (p>0.05) difference in forage crude protein concentration between the two harvesting frequency treatment groups, although the concentration in the group with three times of harvesting frequency tended to be higher. In contrast, crude fiber concentration in the group with two times of harvesting frequency tended to be higher, although the difference was not statistically significant (p>0.05). Crude protein, ether extract, crude fiber, and organic matter concentrations were not significantly (p>0.05) different between the two groups with different fertilization levels. Based on these results, it was concluded that the group with two times of harvesting frequency with conventional fertilization level might be proper for obtaining better forage productivity for Alpine grassland at 800 m altitude.
One of the critical problems to preserve books and documents in libraries and archives is the deterioration. Some of previous results showed that the major cause of paper deterioration was the acid-catalyzed hydrolysis of the cellulose in paper fibres and aging rate of acidic paper was faster than that of alkaline paper. Therefore, It is necessary to remove the acid in the paper for reducing the rate of paper deterioration. It has been reported to extend the useful life of acidic paper by three to five times. Recently, It has been recognized the need for an effective method of deacidifying large quantities of books and document. However, in the previous many reports little attention was paid to the effect of paper additives. In this paper, We carried out experiment about the effect of additives on paper aging and the effect of deacidification by the gaseous ethanolamines (monoehtanolamine, diethanolamine, triehtanolamine). In result, it was found that the strength of aging was in the order of the alum+rosin>alum >AKD> control and the rate of deacidification was in the order of the monoethanolamine>diethanolamine>triethanolamine. The treatment with the gaseous ethanolamines caused decreasing of brightness and dropping of fold endurances. However, deacidification by combination treatment of the various gaseous ehtnaolamines prevented from decreasing of brightness and dropping of folding endurances.
Journal of the Korea Academia-Industrial cooperation Society
/
v.19
no.2
/
pp.56-63
/
2018
The management of Combined Sewer Overflows(CSOs) and Separated Sewer Overflows(SSOs) discharge directly to the effluent system in an untreated state, which occurs when the facility capacity is exceeded due to heavy rain, has become an important issue in recent years as the heavy rain becomes a regular phenomenon. Despite the continuous development of filtration technology, targeting densely populated urban areas, CSOs are rarely applied. Therefore, this study was carried out to optimize the process to apply CSOs in a pilot-scale horizontal flow filtration system with a rope-type synthetic fiber. The research was carried out in two steps: a preliminary study using artificial samples and a field study using sewage. In the preliminary study using an artificial sample, head loss of the filter media itself was analyzed to be approximately 1.1cm, and the head loss was increased by approximately 0.1cm as the linear velocity was increased by 10m/hr. In addition, the SS removal efficiency was stable at 81.4%, the filtration duration was maintained for more than 6 hours, and the average recovery rate of 98% was obtained by air backwashing only. In the on-site evaluation using sewage, the filtration duration was approximately 2 hours and the average removal efficiency of 83.9% was obtained when belt screen (over 450 mesh) was applied as a pre-treatment process to prevent the premature clogging of filter media. To apply the filtration process to CSOs and SSOs, it was concluded that the combination with the pre-treatment process was important to reinforce the hydraulic dimension for the stable maintain of operation period, rather than efficiency. Compared to the dry season, the quality of incoming sewage was lower in the rainy season, which was attributed to the characteristics of the drainage area with higher sanitary sewerage. In addition, the difference in removal efficiency according to the influent quality of the wet season and dry season was small.
Journal of the Korean Society of Fisheries and Ocean Technology
/
v.14
no.2
/
pp.63-68
/
1978
A boat seine has been used as a major fishing gear for catching anchovy (Engraulis japonica) in the southern coastal waters of Korea since the 1920s. Since the 1950s some improvement from the original seine has been made; powered boats equipped with net hauler has been used instead of rowing boats with hand-driven capstan, and the seining method has been changed into the trawling method. But even now, there are many problems to be solved in the view point of decreasing man power without decreasing catching efficiency. For the purpose, patti-net has been introduced from Japan and experimented on the commercial base since 1972, and it was known that the patti-net could be operated with man power as half as needed in the coventional net, but catching efficiency was not so desirable. Therefore, the study on the characteristics of it were required. The authors carried out a model experiment with a Qne-twentieth scale model net towed by a powered boat on the sea. The obtained results run as follows: 1. Hydrodynamic resistance of the model net can be explained as $R_p=69.6 V_{I.66}$$R_h=37 v^2$ where $R_p$ and $R_b$ denote the resistance of the whole gear and the cod end in kg respectively, and v the towing speed in mlsec. 2. Performance of wing and cod end showed no deformation such as observed at the conventional net. 3. The ratio of opening at the entrance of bag net to that of cod end showed about 2: 1. Therefore, when we intend to enlarge the net to be able to operate in the deep fishing ground, the cod end should be enlarged in the same proportion and increased towing power is needed .. Then, it will be better to increase the ratio for increasing fishing efficiency without increasing towing power.
Jo, Beom-Ho;Seol, Min-A;Shin, Su Young;Kim, Il Ryong;Choi, Wonkyun;Eum, Soon-Jae;Song, Hae-Ryong;Lee, Jung Ro
Journal of Plant Biotechnology
/
v.43
no.1
/
pp.91-98
/
2016
The growth area of living modified (LM) cotton has steadily increased every year, since its first commercialization in 1996. Development of environmental risk assessment tools and techniques for LM cotton is required for ecosystem safety. We therefore developed multiplex PCR assays for simultaneous detection of two (MON15985, MON531) and four (GHB614, LLCOTTON25, MON88913 and MON1445) LM cotton events approved in Korea, with event specific primer pairs. The PCR reactions were optimized by using event specific primers of six LM cottons at various concentrations. The reactions allows amplification of estimated amplicons of MON15985 (214 bp), MON531 (270 bp), GHB614 (119 bp), LLCOTTON25 (164 bp), MON88913 (276 bp), and MON1445 (389 bp) from multiplex PCR reactions. The multiplex PCR assay developed allowed that two annealing steps (15 cycles at $55^{\circ}C$ and 25 cycles at $60^{\circ}C$) were performed for amplification of distinguished two LM cottons, and only one annealing step (50 cycles at $60^{\circ}C$) was necessary for tetraplex PCR. Primer extension step of all PCR reactions was skipped for time-effective amplification. Our methods suggest that two multiplex PCR assays can be cost-effective and a rapid diagnostic tool for environmental LMO monitoring of six LM cottons.
Background: As determined from the recent investigations of discordant cardiac xenotransplantation, hyperacute rejection occurs mainly at the endothelial cells in donor microvascular systems, but this does not occur at cardiac valve leaflets or at medium-to-large caliber vessels. On the basis of this background, this study was performed to look into the biocompatibility for transplantation of a middle or large diameter xenogenic blood vessel by conducting xenogenic arterial transplantation with the carotid artery in a pig-to-goat model. Material and Method: The experimental group was composed of 10 pairs of pig-to-goat combinations. They were divided into each period of 1 week, and 1, 3, 6 and 12 months. Four carotid artery grafts obtained through collection of the bilateral carotid arteries from two pigs were preserved at $-70^{\circ}C$ without other treatment, and then they were transplanted into the bilateral carotid arteries of two goats. Doppler ultrasonography was done on a periodic basis after transplantation to evaluate the patency of the grafted blood vessel. At the ends of a predetermined period, the grafts were explanted from the goats and they underwent gross examination. Hematoxylin-eosin and Masson's trichrome staining were conducted. In addition, in order to examine the immunological rejection of the grafted xenogenic blood vessel, immunohistochemical staining was conducted with T-lymphocyte indicator and von Willebrand factor. Result: Two goats at the each one-week period and the one-year period died during the experimental period because of a reason unrelated to the experimental procedure, and the remaining 8 goats survived until the end of each experiment period. On Doppler ultrasonography, unilateral carotid artery occlusion was found in a goat, whose period was specified as 3 months, among the 8 survived goats. However, the vascular patency was maintained well and there was no graft that formed aneurysms in the other goats. On gross examination, the region of vascular anastomosis was preserved well, and calcification of the grafted blood vessel was not shown. Histologically, the endothelial cells of the graft disappeared one week after transplantation, and then there was progressive spread of the recipients' endothelial cells from the anastomotic site. The reendothelialization occurred over the whole graft at one month after transplantation. The neointimal thickening and adventitial inflammation became severe by 3 months after transplantation, but this lessened at 6 months and 12 months, respectively. The rate of CD3 positive cells was very low among the infiltrated inflammatory cells. Conclusion: The fresh-frozen xenogenic artery kept its patency without being greatly influenced by xenogenic immune reaction.
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