The exogenous gene transfer by intracytoplasmic sperm injection (ICSI) procedure has been recently used to produce transgenic mice and pigs. Sperm-mediated DNA transfer has the potential to markedly simplify the generation of transgenic animals. This method may serve as an alternative to the pronucleus injection of DNA for the production of transgenic pigs. Therefore, in this study, we investigated the expression of transgene after co-injection of spermatozoon or sperm head with green fluorescent protein (GFP) gene into in vitro matured porcine oocytes. Spermatozoon and sperm head, that was obtained by sonication, were treated with 0.03% Triton X-100 to remove the membrane. They were preincubated with linearized pEGFP-N1 for 1 min, and then embryos cultured NCSU23 medium for 2.5 days after co-injected of sperm and DNA. We monitored expression of GFP in embryos under epifluorescent microscope. The remove of sperm membrane did not alter the developmental competence of embryos after ICSI. At 7 days following injection, the rates of blastocysts following injection of intact sperm (15.0%), and of sperm with disrupted membrane (14.2%) were higher than that following IVF (10.0%). Porcine oocytes injected with sperm which co-cultured with DNA concentration of 1, 0.1, and 0.01 ng were 60, 65.7 and 75% and 18.5, 37.4 and 22.2% for rates of cleavage and GFP expression, respectively. In vitro matured porcine oocytes injected with sperm and isolated sperm head resulted in 69 and 59.7% of cleavage rates, respectively The rates of embryo GFP expressed did not significantly different between sperm (20.4%) and sperm head (20.0%) injection. The transgenic embryos with the clusters of positive blastomeres were observed under fluorescent microscope. Most of embryos expressed GFP gene showed mosaicism. They showed GFP expression at 1/4, 2/4 and 3/4 of blastomeres at the 4-cell stage. Among these 4-cell embryos, the expression rate of 1/4 blastomere group (54.6%) was higher than the other groups (15.3-30.7%). These results indicate that membrane disrupted sperm could attach with exogenous DNA, and that this procedure may be useful to introduce foreign gene into porcine oocytes. Therefore, our data suggest that the ICSI car be a useful tool to efficiently produce transgenic pig as well as other mammals.
This study was conducted to investigate liquid pig manure (LPM) application rates on the growth characteristics, yield, and feed value of whole crop barley in Yeongsangang and Saemangeum reclaimed tidal land. Electronic conductivity (EC), organic matter (OM), and available phosphate (Av. $P_2O_5$) increased in chemical properties of Yeongsangang and Saemangeum soil as raising LPM application level. As increasing LPM application level, exchangeable $Na^+$ significantly increased in Yeongsangang, while exchangeable $K^+$ significantly increased in Saemangeum. Plant height was not significantly different from LPM 100% to LPM 200% in Yeongsangang and in Saemangeum. Dry matter yield of whole crop barley increased steadily, but crop yield of LPM 200% in Yeongsangang ($10.5ton\;ha^{-1}$) was as much as that of LPM 150% ($10.0ton\;ha^{-1}$). Yield of LPM 200% ($11.2ton\;ha^{-1}$) in Saemangeum was similar to that of LPM 150% ($10.5ton\;ha^{-1}$). Crude protein (CP) increased depending on LPM application level, but total digestible nutrients (TDN) increased regardless of LPM application level. LPM 200% was the highest in TDN yield (Yeongsangang: $7.4ton\;ha^{-1}$, Saemangeum: $6.9ton\;ha^{-1}$), but there was no statistical difference between LPM 150% (Yeongsangang: $6.9ton\;ha^{-1}$, Saemangeum: $6.6ton\;ha^{-1}$) and LPM 200%. From the results described above, optimum rate of LPM for cultivating whole crop barley is considered 100% in Yeongsangang reclaimed tidal land and 150% in Saemangeum reclaimed tidal land, showing that the effect of LPM application is better in Segmentation than that in Yeongsangang for yield of whole crop barley.
Cha, Ju-Su;Kim, Sun Hyo;Jung, Samuel;Kang, Ho Jin;Jo, Cheorun;Nam, Ki-Chang
Korean Journal of Poultry Science
/
v.41
no.1
/
pp.53-59
/
2014
To evaluate the meat quality differences of Korean Native Chickens (KNC) available in Korean market, the physicochemical and sensory characteristics of a broiler and two KNC (HH and WD breed) were analyzed. The fat content of KNC WD breast meat was higher than that of KNC HH. The breast and thigh meats of KNC HH had higher $L^*$ and $a^*$ values than the broiler or the KNC WD. WD meats showed greater DPPH radical scavenging activity and higher pH values than the HH. For the fatty acid composition, there was no significant difference in total content of the saturated fatty acids among chicken breeds (p>0.05). KNC had greater content of polyunsaturated fatty acids, especially arachidonic acid (20:4) and DHA (22:6), compared with the broiler. KNC WD meats had higher composition of linolenic acid (18:3) than the KNC HH. For sensory evaluation, the WD breast meat showed higher taste and tenderness scores than the HH. On the other hand, the soup made by broiler had higher taste scores than the KNC. The breast and thigh meat from two KNC breeds showed different quality parameters that may come from the differences of genetic attributes, feed, and production strategy between two breeds.
Kim, Na-Kyeong;Park, Jung-Min;Lee, Jung-Hoon;Kim, Ha-Jung;Oh, Sejong;Imm, Jee-Young;Lim, Kwang-Sei;Kim, Jin-Man
Food Science of Animal Resources
/
v.35
no.1
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pp.143-148
/
2015
Yogurt powder is fermented milk processed in the form of dry yogurt, and has advantages such as stability, storability, convenience, and portability. China and Vietnam are important export target countries because of the increased demand for dairy products. Therefore, we surveyed dairy product standardization in order to establish an export strategy. Lactic acid bacteria counts are unregulated in Korea and Vietnam. In China, lactic acid bacteria counts are regulated at $1{\times}10^6$ colonyforming units (CFU)/mL and detected at $6.24{\pm}0.33\;Log\;CFU/mL$. All three countries have regulated standards for total bacterial counts. In China, total bacterial counts of milk powder are regulated to n=5, c=2, m=50,000, M=200,000 and detected at $6.02{\pm}0.12\;Log\;CFU/mL$, exceeding the acceptable level. Lactic acid bacterial counts appeared to exceed total bacterial counts. Coliform group counts, Staphylococcus aureus, Listeria monocytogenes, and Salmonella species were not detected. Acidity is not regulated in Korea and Vietnam. In China, acidity was regulated to over $70^{\circ}T$ and detected $352.38{\pm}10.24^{\circ}T$. pH is unregulated in all three countries. pH was compared to that of general fermented milk, which is 4.2, and that of the sample was $4.28{\pm}0.01$. Aflatoxin levels are not regulated in Korea and China. In Vietnam, aflatoxin level is regulated at 0.05 ppb. Therefore, all ingredients of the yogurt powder met the safety standards. This data obtained in this study can be used as the basic data in assessing the export quality of yogurt powder.
Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.
This study was carried out to develop salt tolerant varities of rapeseed (Brassica napus L.) which can be grown in the high salty reclaimed land. The seeds of three varieties 'Naehan', 'Tammi', and 'Halla' were treated by proton ion beams and gamma rays with 0 to 2,000 Gy. For the selection of salt tolerant lines, emergence and survival rate, and growth characteristics of $M_2$ to $M_4$ generations were investigated in the Saemangeum reclaimed fields with the different salt concentrations. The lines with potential salt tolerance were selected in the $M_4$ generation and tested indoor for their growth characteristics. There was no significant changes in the soil pH for $M_4$ generation during growth period. However, soil EC was higher in early spring than sowing period (mid October). In $M_4$ generation test, the seeds of original and selected line showed high rates of emergence and survival, as determined one month after sowing. After wintering, however, the original varieties showed the significant reduction in the survival rate, while the selected lines showed a higher survival rate and good growth, leading to the completion of their life cycle. Consequently we selected 9 lines from $M_4$ generation with better performance in growth and yield. Soil EC was $2.8{\sim}4.3dS{\cdot}m^{-1}$ during $M_4$ generation growth period. The laboratory test of the lines selected from $M_4$ generation was made for their salt tolerance potential. The selected lines showed higher chlorophyll and proline contents than the original varieties. There was also no significant difference in the emergence rate of seed between the original and selected varieties. In 200 mM natural sea salt, the $N{\gamma}600-21-1-641$ line derived from 'Naehan' was the highest in growth rate, leaf chlorophyll and proline contents. $T{\gamma}800-20-2-461$ line derived from 'Tammi' didn't show significant difference in growth rate compared to original variety in 200 mM and withered in 250 mM like other lines as time passed. $H{\gamma}200-7-1-740$ line showed similar growth and chlorophyll content compared to its original variety.
The Hadano Basin is located at a distance of about 70kms and 60kms from Tokyo and Yokohama and lies in the south-west part of the Kanto region in Japan. The basin area, which correspoends to the catchment of the Kaname River, is about areal size of 60.7$\textrm{km}^2$ and extends about length of 8kms in E-W direction and about width of 5kms in N-S direction (Fig.1). The Hadano basin is filled with thick pile of the alluvum from deposits composed of volcanic materials, mostly came from the Hakone Volcano and overlain by Fuji Volcanic ashes. Fluvial deposits form the good aquifer, therefore water resources of Handano City has been largely depending upon the eroundwater. Urbanization and industrialization of the basin has been rapid in the last thirty years, after activation of "Factory Attraction Policy of Hadano City" in 1956. Growth in population and number of factory due to urbanization changed the land-use pattern of the basin rapidly and increased the water demands. Therefore, Hadano City exploited a new source of water supply, and have introduced the prefectureal waterworks since 1976. On the other hand, the rapid urbanization has brought about the pollution of streams in the basin by domestic sewage and industrial waste water. Diffusion rate of sewerage systems in Hadano City is 38% in 1993. In ordcr to examine the impact of anthropogenic factors on river environments, the author took up the change of land-use and diffusion area of sewerage as parameters, and performed field surveys on water discharge and quality. The survey has been made at upstream and downstream of the main stream regularly per month, to get informati ons about the variation of discharge and water quality aiong the stream and its diurnal fluctuation. Annual variation has been analyzed based the data from Hadano City Office. The results are summarized as follows. 1. Stream discharge has been increasing by urbanization (Fig.3). Water quality (C $l^{-10}$ , N $H^{+}$$_{ 4}$-N, BOD) has been improving gradually after the application of sewerage service, yet water pollution load at the lower station has increased than that at the upper one because of the larger anthropogenic discharge volumes (Fig.4). 2. Corrclation coefficient of discharges between upper and lower was 0.81-0.92. Pollutant loads of the R. Kamame after the confluence with R. Kuzuha grew up by 2.4-3.7 times as compared with its upper reaches, and it increased to 3.7-6.9 times after the confluence with the R. Muro (Fig.5). 3. The changes of water quality along the stream can be divided into two groups (Fig.6a). First: water quality of the R. Kaname and R. Shijuhachisse is becoming worse towards the lower reaches because the water from branches are polluted. Second: water quality are improved in the lower where spring and small branch streams supply clear water, for example R. Mizunashi, R. Muro and R. Kuzuha. 4. Measured discharge at the upper station in the R. Shijuhachisse is 0.153㎥/sec, and about 55% of this is recharged until it reaches to the lower point. The R. Mizunashi has a discharge of 1.155㎥/sec at the upper point, is recharged 0.24㎥/sec until the midstream and groundwater spring 0.2㎥/sec at the lower reaches. R. Kuzuha recharged all the mountain runoff (0.2㎥/sec) at the upper reaches. The R. Muro is supplied by many springs and the estimated discharge of spring was 0.47㎥/sec (Fig.6b). 5. Diurmal variations in discharge and water quality are influenced clearly by domestic and industrial waste waters (Fig.7, 8).ed clearly by domestic and industrial waste waters (Fig.7, 8).
Pressurization walk training (PWT) with blood flow occlusion has been investigated with regard to muscle hypertrophy and physical fitness function in athletes and healthy people. However, the cardiorespiratory and cardiovascular responses of obese people to PWT are unknown. Thus, we investigated the effects of PWT on body composition (Weight, FM, LBM, %fat, BMI), cardiovascular responses (HR, SV, CO, TVC), and cardiorespiratory responses ($VO_2max$, VEmax, HRmax) in middle-aged obese women. They participated in walk training with (n=15) blood flow occlusion and cross-sectional areas of the quadriceps on both legs. Five sets of 3-min walking (5.5 km/h at 5% grade) and 1-min resting were performed twice a day, 5 days/week for 3 weeks. The results showed that the LBM was significantly increased, and decreased body weight of reducing FM, %bodyfat in PWT ($p$<0.05). For the cardiovascular response, SBP and TPR were significantly decreased ($p$<0.05), and CO increased ($p$<0.05). In addition, the $VO_2max$ and VEmax were improved through PWT. Therefore, this study suggests that the presence of obesity in middle-aged women may result in body composition, cardiorespiratory, and cardiovascular responses caused by PWT.
The physico-chemical characteristics of water and sediment, and structures of vegetation of the vascular hydrophytes were investigated in the West Nakdong River. Water quality was eutrophic according to the mean values and the ranges of water properties such as pH, DO, BOD, chlorophyll a, total nitrogen and phosphate, and other nutrients. A few cases were hypereutrophic for chlorophyll a level in summer. Soil reaction was weak acid. Composition of sediment was mainly sand except in SI(Sinan chideung) of which was mainly clay, and SU(Suan chideung) of which was mainly silt. Flora of vascular hydrophyte had 26 species and 1 variety comprising 16 families. Trapa japonica was dominant species in the sites of DU(Dunchido), GA(Garak chideung) and SU. Nymphoides peltata and Hydrocharis dubia dominated in DA and SI, respectively. Species diversity and evenness were relatively high in SI and SU but dominance was high in DA. After June, water lettuce(Pistia stratiotes) and water hyacinth(Eichhornia crassipes) were flowed from tributary to the river. Standing crop of macrohydrophytes was high in DA from April to August, but it showed maximum standing crop (445g·dw/㎡) in DU after disturbance by explosive growth of exotic plants in October. In comparison with those in 1985, total productivities in DU and GA decreased to 33.5%, and the reduction ratio of dominant species, Trapa japonica was 56.7%. Najas marina, N. minor, Myriophyllum spicatum and Nymphoides indica have disappeared ever since the Nakdong barrage was constructed in the Nakdong river. They were divided into three groups (GA-SU-DU, DA, SI) by cluster analysis. Introduction of the exotic species in this river caused decreasing of endemic plants including endangered species Euryale ferox and rare species Hydocharis dubia, and food plants for waterfowl such as Trapa japonica, Vallisneria asiatica and Potamogeton crispus.
This study was conducted to prepare high quality grape seed oils by solvent extraction and chemical refining process. Additionally, quantitative analysis of several functional components in grope seed was carried out to compare quality characteristics of grape seeds from grapes grown by conventional and organic agricultural practices. There are no significant differences in several functional constituents of grape seeds between conventionally cultivated- and organically cultivated-grapes, although some functional compositions of grape seeds are different between two cultivation methods. The dried grape seed was pretreated with roasting heating for 5 min, milled and then extracted twice with n-hexane under reflux at $50^{\circ}C$ for overnight, followed by filtration and evaporation. The crude grape seed oil was successively purified by degumming with $0.1\%\;H_3PO_4$, deaciding with $20\%\;NaOH$, and then decoloring and deodorization by a steam distillation, and thereby producing purified grape seed oil(yield: $5.0\%/dried$ grape seed). Physicochemical characteristics of the purified grape seed oil were comparable to those of the imported grape seed oils.
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