Park, S. W.;M. R. Shin.;Kim, Y. H. .;H Shim;Kim, N. H.
Korean Journal of Animal Reproduction
/
v.25
no.1
/
pp.51-61
/
2001
This study was conducted to determine developmental ability of reconstructed embryos by nuclear transfer using somatic cell of Korean bull with high genetic value. Fibroblast cells obtained from ear biopsy of the bull were cultured in Dulbecco's Modified Eagle's medium (DMEM) at 37$^{\circ}C$ in air containing 5% $CO_2$. The cummulus-oocyte complexes were collected from slaughterhouse and were matured in vitro for 20 h in TCM 199 culture medium and the oocytes were then enucleated in modified phosphate buffered saline with cytochalasin B. Matured bovine oocytes were enucleated by aspirating the first polar body and metaphase chromatin using a beveled pipette in modified phosphate buffered saline. The ear fibroblast cells were fused into enucleated oocyte by electrical stimulation. The reconstructed oocytes were activated with ionomycine and 6-dimethylaminopurine, and then cultured in CR1aa medium for 7.5 days. Out of 524 bovine eggs reconstructed by nuclear transfer 65.6%(277/422) embryos were cleaved, and 30.7% (85/277) cleaved embryos were developed to the morula to blastocysts. There was no difference of developmental ability in vitro of reconstructed embryos regardless of donor cell passages. In order to determine fate of foreign mitochondria of donor nucleus, the Mito Tracker stained cells were fused into enucleated oocytes. The donor mitochondria were detected early stage of embryos, but disappeared rapidly. The developmental ability of reconstructed embryos was not impaired by Mito Tracker treatments. The results indicate that viable reconstructed embryos can be producted by nuclear transfer using somatic cell of Korean bulls.bulls.
Shin J. S.;Kim, C.;S. J. Uhm;Kim, N. H.;Lee, H. T.;K. S. Chung
Korean Journal of Animal Reproduction
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v.22
no.4
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pp.395-403
/
1998
In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence and absence of artificial activation. Electrical stimulation at 2 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocyto chemistry and laser scanning confocal microscopy study revealed that microtubuels were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. At 6 days following injection blastocoele formation was seen in the eggs following round spermatid (25%) and round spermatid nucleus injection (27%). However, none of oocytes developed to the blastocyst stage at 6 days following sham injection. The average cell numbers of blastocysts at 8 days following injection of spermatid and spermatid nucleus were 87 to 99. These results suggested that either round spermatid or it's nudeus can be used to produce viable embryos by injection into unfertilized oocytes in the pig.
For a long-term preservation of silkworm stocks by frozen gonad storage, fundamental topics such as freezing rate and transplanting stage of the gonad, proper cryoprotectant, and super-cooling temperature and freezing point of the freezing medium were examined and following results were obtained. Proper method to anesthetize the ovary-recipient silkworm was to dip the animal to cold water for 10 minutes, and the ovary taken from the 4th instar larvae was more suitable for freezing-preservation than that from the 5th. Concerning the cryoprotectant, glycerol and DMSO were effective to prevent cryoinjury of the ovary, but sorbitol was not. The supercooling temperature and freezing point of the medium to freeze the ovary and testes were checked, and consulting with the results desirable cooling rate was confirmed. On the desirable conditions of transplanting methods, freezing rate and cryoprotectant concentration ect., the next generation was obtained when the females implanted frozen-thawed ovaries mated with normal males, but none of the normal females mated with the males implanted frozen-thawed testes laid fertilized eggs. Now it is needed to improve the connecting ration of the ducts associated with the transplanted testis to those of the hosts.
Intraspecific diploid androgenesis was achieved in mud loach (Misgurnus mizolepis) by the inhibition of the first mitotic division using combined thermal treatment. A combined thermal treatment (heat shock at $40.5\;^{\circ}C$ for 120 sec followed by cold treatment at $1\;^{\circ}C$ for 45 min) applied to the 1st metaphase of cell division (28 min post insemination at $25\;^{\circ}C$) successfully recovered viable androgenetic diploidy. Mean hatching success of the androgenetic diploid group was 29.6%, and the average yield out of total eggs taken was about 7% assessed at 1 week of age. However, relatively large variations in the yield of diploid androgenesis were observed among different egg batches used as cytoplasmic donors. Successful diploidization was confirmed by flow cytometric analysis, and parthenogenic reproduction in a paternal exclusive manner was verified with transgene dosage. Significant mortality was found in most androgenetic groups especially from hatch to 1 month of age, although such mortality was stabilized later.
Selection of breeds, adequate environment and sanitary condition must be considered to improve production ability. Minimizing of production cost through reasonable management and farm system can introduce maximizing net income. Followings are the summary of poultry industry in Japan: (1) To decreasing consumers Price and to gurantee the quality of eggs, poultry farms changed their farm system as complex which contain feed factory, GP center and processing factory as well as raising laying hens. (2) Broiler farms moved from suburbs to several remote from cites owing to development of transferation , and rising price of land. (3) The egg Price in Japan was depended on season. From fall to spring, the Price was usually rised due to increased consumption, and from May to August the price always dropped. (4) The average production per one you was around 17kg/hen and average feed consumption was around 2.45. Rearing rate was around 98.5% and motality of hens was around 7%. And one person reared 15,000-20,000 hens in over 100 thousand chicken reared farms even though average ability per person showed 3,266 hens.
The main purposes of the nutrition and clinical surveys were to provide baseline information on the nutritional status of pre-school children in rural health demonstration project areas of the Korea Health Development Institute (KHDI) for nutrition guidance services for the MCH target group. The survey covered a total of 222 pre-school children and 135 mothers in Okgu Gun, Cholla Pukto Province from August 10 to August 17, 1979. The survey results are summarized as follows: 1) Family Environment Seventy percent of the households had more than three children, and the mean family sire was 6. Sixty-nine percent of the mothers and 47% of the fathers of the surveyed households were educated at or below the primary school level. The majority, 70% of the mothers, were aged between 20 years and 35 years. 2) Anthropometric Measurements and Hemoglobin Value 4.5% of the children were lower than 80% weight for age of the Korean standard, and 5.4% were lower than 85% arm circumference for age of the Jelliffe's standard resectively, and those were suffering from protein-energy malnutrition. Angular stomatitis were observed on 66.2% of the subjects. Mean hemoglobin value was 11.1g/100m1, and 44.2% of the subjects were categorized as anaemia. 3) Food and Nutrient Intake of animal foods was very low, ranging from 2.9 to 17%. Consumption of eggs was less than 2% of total food intake, and intake of legumes was also very scanty, between 0.8 to 3.7%. These data present evidence of very poor protein intake, quality as well as quantity. Energy intake of children was 60.0 to 64.4% of the recommended allowance, and mean protein intake only met 47.4% of the recommendation. Low intake of vitamins except thiamin were also found. 4) Mother's Nutrition Knowledge Eighty-five percent of the mothers were entirely ignorant regarding the 'five basic food group' which is most important fact on food and nutrition guidance. Mean knowledge score from 14 basic questions about food and nutrition was as low as 5.1. There was a significant positive correlation between mother's educational level and nutrition knowledge score. 5) Family Planning Variable There were significant correlation among maternal, family planning variables, and some of the nutritional and physical measurements. The study revealed that the mother's educational level and nutrition knowledge score are more crucial factors than the family planning variables on effecting food intakes on children. Recommendation : According to the results of the surveys, there were high incident rates of nutritional anaemia and angular stomatitis among pre-school children, and most of rural women had very limited knowledge about food and nutrition. As a main part of the health education activities, the community health workers should provide nutrition education to the village mothers to improve the nutrional status of young children in rural areas. Nutrional promotion at the primary health care level should be mainly based on appropriate nutrition education.
Purpose: The purpose of this study was to estimate dietary folate intake and food sources of children and adolescents using a nutrient database revised based on measured folate in selected foods. Methods: Folate content in 51 foods known as folate sources was measured by microbiological assay after trienzyme extraction. Folate intake was estimated from a part of the data of 'Dietary Intake Survey of Infants, Children and Adolescents in 2007~2008' conducted by the Korea Food and Drug Administration (KFDA) and the Korea Health Industry Development Institute (KHIDI). The study subjects were 567 children aged 1~19 years living in the Chungcheong area who completed two 24-hour recalls. Results: Folate values were revised by replacing the values in the current database with the analyzed values except when the value in the current database was between the analytical values or was not different from the mean analytical value by more than 10%. Among the revised values of 40 food items, folate values of 36 foods were lower than the current values. Mean folate intakes estimated with the revised database were approximately 70~80% lower than those estimated with the current database. Mean folate intakes of males aged 12~14 and females aged 12~19 were lower than the 2010 Recommended Nutrient Intakes (RNIs). Chicken's eggs, Kimchi, rice, mandarin, and laver were found to be main food sources of folate. Conclusion: In this study, mean dietary folate intakes were lower than those estimated with the current database. Further analyses for folate content especially in cooked foods commonly consumed in Korea are needed using a reliable assay in order to accurately assess folate intake of the Korean population. In addition, nutrition education should be provided for adolescent females in order to increase consumption of folate-rich foods.
The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.
Recently, we found and analyzed vitamin $B_{12}$ in some Korean traditional plant foods which had not reported, yet. This study was to investigate vitamin $B_{12}$ intake and its dietary sources and the vitamin $B_{12}$ status in the very old elderly Koreans. We measured serum vitamin $B_{12}$ level and estimated the amounts of vitamin $B_{12}$ intake from different dietary sources in female elderly Koreans aged 85 and over who had consumed a relatively low animal traditional diet for the whole life. The average age of the subjects (n = 127) was 98.0 years (85-108 years). The assessment on energy and nutrient intake involved a one-day 24-hour recall, and serum vitamin $B_{12}$ concentration was measured by radioimmunoassay. Overall diet pattern was not different between the 85-99 yr-old group and centenarians, except centenarians were taking more dairy product. The average ratio of plant food to animal food consumption was 87.5:12.5 in weight. The average vitamin $B_{12}$ intake of our subjects was 3.2 ${\mu}g$, and 52.7% of subjects consumed under estimated average requirement, 2.0 ${\mu}g$/day. On dietary source, 67.3% of dietary vitamin $B_{12}$ was from meat, eggs and fishes and 30.6% was from plant foods, such as soybean-fermented foods, seaweeds, and kimchi. The average serum vitamin $B_{12}$ concentration was 450.5 pg/mL, and low serum vitamin $B_{12}$ (< 200 pg/mL) was found in 9.6% of subjects. Dietary vitamin $B_{12}$ intake was significantly lower in subjects with low serum vitamin $B_{12}$ (0.79 ${\mu}g$/day) than those with normal serum vitamin $B_{12}$ (3.47 ${\mu}g$/day). There were no significant difference in vitamin $B_{12}$ intake and its dietary sources and serum vitamin $B_{12}$ level between the 85-99 yr- old group and centenarians. In conclusion, several plant-origin foods including seaweed, soybean-fermented foods, and kimchi, may contribute significantly to good vitamin $B_{12}$ status in very old elderly Koreans.
This study was aimed for development of a useful genes that has a transcript expressional specificity in the early embryonic stage of the silkworm, Bombyx mori. We constructed and analyzed a full-length cDNA library from silkworm's eggs which after a lapse of 2 ~ 6 hours post oviposit. A total 960 clones were randomly selected, and the 5' ends of the inserts were sequenced to generate 652 expressed sequence tags(EST). 334 unique ESTs were generated after the assembly of 652 ESTs. The annotation of 334 unique ESTs by BLAST search revealed that 156(47%) of the sequences represented known genes, whereas 178(53%) of the sequences has no matches in the database. Of the 156 known genes, the most abundant genes were heat shock protein hsp20.8 gene(12 times) and ubiqutin-like protein gene(11 times). The functional groups of these ESTs with matches in the database were constructed according to their putative molecular functions. Among thirteen functional categories, the largest groups were protein synthesis(9.6%) and cellular organization( 8.1%). Further defined studies on molecular functions and biological roles of their promoters will give us wellfined information and its application.
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