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In vitro Development of Somatic Cell Nuclear Transferred Bovine Embryos Following Activation Timing in Enucleated and Cryopreserved MII Oocytes (탈핵 후 동결한 MII 난자의 활성화 시기가 체세포 핵치환 이후 소 난자의 체외발달에 미치는 영향)

  • 박세필;김은영;김선균;이영재;길광수;박세영;윤지연;이창현;정길생
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.245-252
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    • 2002
  • This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.

Paradoxical Upper Airway Obstruction and Central Sleep Apnea Developed After Anterior Cervical Spine Fusion (전방경추융합술 후 발생한 역설상기도폐쇄 및 중추성 수면 무호흡)

  • Lee, Sang Haak;Choi, Young Mee;Park, Ye Ree;Kang, Ji Ho;Kim, Young Kyoon;Kim, Kwan Hyoung;Song, Jeong Sup;Park, Sung Hak;Moon, Hwa Sik
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.3
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    • pp.295-298
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    • 2005
  • We report a case of a 48-year-old man with a paradoxic upper airway obstruction and central sleep apnea that developed after an anterior cervical spinal fusion. Nine months before being admitted to this hospital, he was diagnosed with a herniated intervertebral disc between the 5th and 6th cervical spine, and the first operation was carried out. Two months later, a pseudoarthrosis has developed and a second operation, an anterior interbody fusion of the C5 and C6 using autogenous strut bone graft, was performed. After the second operation, he began to complain of snoring, excessive daytime sleepiness, insomnia, and a bizarre sound heard near the upper airway during breathing. Nasopharyngoscope and magnetic resonance imaging disclosed a paradoxical narrowing of the nasopharynx during expiration. On the overnight polysomnography, the apnea index was 8.7/h (central apnea, 7.0/h; obstructive apnea, 1.7/h). Nasal continuous positive airway pressure was applied, but he complained of pressure-intolerance, and laser-assisted uvulopalatoplasty was then performed. Two months after surgery, clinical symptoms as well as the apneas had improved markedly. We suggest that this paradoxic upper airway obstruction might be associated with the anterior cervical spinal surgery even though the mechanism is unclear. This case also emphasizes that an upper airway obstruction can contribute to the development of central sleep apnea.

Quality Characteristics of Saccharified Rice Gruel Prepared with Different Cereal Koji (곡류 코지를 이용하여 제조한 당화쌀죽의 품질 특성)

  • Hwang, In-Guk;Yang, Ji-Won;Kim, Ja-Young;Yoo, Seon-Mi;Kim, Gi-Chang;Kim, Jin-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1617-1622
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    • 2011
  • We conducted this study to investigate the quality characteristics of saccharified rice gruel prepared with different cereal koji mold (rice, buckwheat, sorghum, adlay, or Italian millet koji). The moisture, crude protein, crude lipid, and crude ash content of rice, buckwheat, sorghum, adlay, and Italian millet showed a range of 11.12~12.85, 5.81~16.24, 0.56~4.36, and 0.28~1.93%, respectively. The proximate composition of saccharified rice gruel prepared with different cereal koji was significantly (p<0.05) higher than unsaccharified rice gruel. The pH, L, a, and b values of saccharified rice gruel ranged between 6.09~6.39, 60.38~78.25, -0.87~5.70, and 7.74~13.37, respectively. The viscosity of saccharified rice gruel prepared with different cereal koji was significantly (p<0.05) lower than the unsaccharified rice gruel. After saccharification, the soluble solids, glucose, and maltose content of saccharified rice gruel prepared with different cereal koji were significantly increased in the range of 11.47~13.03$^{\circ}Brix$, 0.60~1.44%, and 0.47~0.88%, respectively. A sensory evaluation indicated that saccharified rice gruel was significantly (p<0.05) better than unsaccharified rice gruel. In conclusion, cereal koji could be used as a gruel processing method to increase the sensory properties and nutritional values of gruels.

Hormonal Changes in Cultured Medium on In Vitro Culture of Bovine Oviduct Epithelial Cells (BOEC) Supplemented with UK-4 (소 난관 상피 세포의 체외 배양시 IL-4의 첨가에 의한 배양액내 호르몬 변화)

  • Choi, S.H.;Cho, S.R.;Han, M.H.;Kim, H.J.;Son, D.S.;Sang, B.D.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.281-286
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    • 2006
  • This study was conducted to investigate the hormonal changes in cultured medium during in vitro culture of bovine oviduct epithelial cells (BOEC) supplemented with interleukin (IL)-4 of 0.001, 0.01, 0.1 or 1 ng/ml. BOEC were collected from the oviduct and washed 3 times with 1% antibiotic-mycotic-DMEM medium and cultured at $39^{\circ}C$, 5% $CO_2$, 95% air for 24$\sim$120 hrs. The cultured media were analyzed hormonal changes with hormonal analyzing kit (progesterone (P4), estradiol (E2) : Perkin Elmer, USA) and Transforming growth factor (TGF)-$\beta$ with Eliza kit (Promega, USA). The production of P4 in 0.001 IL-4 was increased as the culture time increased. P4 production was significantly higher in the medium cultured for 120 hrs than 24 hrs (P<0.05). P4 production in 0.01 ng/ml group was similar to that of 0.001 ng/ml. The production of E2 in 0.001 and 0.01 ng/ml groups were increased to 72 hrs like P4 production and showed significantly different between the culture periods (P<0.05). After the culture for 96 hrs, P4 and E2 production were increased to 96 hrs, but decreased at 120 hrs. The production of TGF-$\beta$ showed no changes according to culture period or supplementation of IL-4. In conclusion, the supplementation of IL-4 can increase the production of P4 and E2 and might have important role for the successful pregnancy in bovine.

Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes III. Effects of Fertilization Media and the Sperm Concentration during Fertilization on In Vitro Fertilization and Development (돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 III. 체외수정배양액과 정자농도가 체외수정 및 체외발달에 미치는 영향)

  • Yeon S. H.;Son D. S.;Jean H. J.;Choi S. H.;Kim I. C.;Park C. S.;Lee K. S.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.265-273
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    • 2004
  • This study was carried out to examine the effects of fertilization media and sperm concentration on in vitro fertilization (IVF) and development (IVD) of porcine oocytes matured in vitro. Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10% porcine follicular fluid (pFF). After the fertilization by experimental scheme, putative embryos were developed in vitro in NCSU-23. The results are as follows. When the oocytes were fertilized in vitro in modified TBM or modified TLP-PVA by 1 ${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were not significantly different between two media. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were not different between two media, either. When the oocytes were fertilized in vitro in mTBM by 5${\times}$10$^4$, 1${\times}$10$^{5}$ or 5${\times}$10$^{5}$ sperm/$m\ell$, all of the fertilization parameters were significantly (P<0.05 or P<0.01) increased as sperm concentration was elevated. Subsequently, as these putative embryos were developed in vitro in NCSU-23, the percentage of oocytes cleaved and of blastocysts were significantly boosted (P<0.01) as sperm concentration at fertilization was elevated from 5${\times}$10$^4$ to 1${\times}$10$^{5}$ sperm/$m\ell$, but were not different between 1${\times}$10$^{5}$ and 5${\times}$10$^{5}$ sperm/$m\ell$.

Characteristics of Phytoplankton Succession Based on the Functional Group in the Enclosed Culture System (대형 배양장치에서 기능그룹에 기초한 식물플랑크톤 천이 특성)

  • Lee, Kyung-Lak;Noh, Seongyu;Lee, Jaeyoon;Yoon, Sungae;Lee, Jaehak;Shin, Yuna;Lee, Su-Woong;Rhew, Doughee;Lee, Jaekwan
    • Korean Journal of Ecology and Environment
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    • v.50 no.4
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    • pp.441-451
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    • 2017
  • The present study was conducted from August to December 2016 in a cylindrical water tank with a diameter of 1 m, a height of 4 m and a capacity of 3,000 L. The field water and sediment from the Nakdong River were also sampled for the experimental culture (field water+sediment) and control culture (field water), respectively. In this study, we aimed to investigate phytoplankton succession pattern using the phytoplankton functional group in the enclosed culture system. A total of 50 species in 27 genera including Chlorophyceae (30 species), Bacillariophyceae (11 species), Cyanophyceae (7 species), and Cryptophyceae (2 species) were identified in the experimental and control culture systems. A total of 19 phytoplankton functional groups (PFGs) were identified, and these groups include B, C, D, F, G, H1, J, K, Lo, M, MP, N, P, S1, $T_B$, $W_0$, X1, X2 and Y. In particular, $W_0$, J and M groups exhibited the marked succession in the experimental culture system with higher biovolumes compared to those of the control culture system, which may be related to the internal cycling of nutrients by sediment in the experimental culture system. The principal component analyses demonstrated that succession patterns in PFG were associated with the main environmental factors such as nutrients(N, P), water temperature and light intensity in two culture systems. In conclusion, the present study showed the potential applicability of the functional group for understanding the adaptation strategies and ecological traits of the phytoplankton succession in the water bodies of Korea.

Conversion of Fermented Feed by Basidiomycetes (담자균(擔子菌)을 이용(利用)한 발효(醱酵) 사료(飼料)에 관한 연구(硏究))

  • Hong, Jae-Sik;Kim, Joong-Man;Jeong, Jin-Chul;Lee, Tae-Kyu;Kim, Dong-Han;Kim, Myung-Kon;Lee, Keug-Ro
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.157-168
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    • 1985
  • To investigate nutritive values of a feed fermented with basidiomycetes, among the isolated strains, Lyophyllum decastes (Fr.) Sing. was found with the greatest enzyme productivity and rapid mycelial growth in rice straw medium. Optimum temperature, pH and moisture content for mycelial growth and enzyme production of the strain were $25{\sim}30^{\circ}C,\;pH\;4.0{\sim}7.0\;and\;70{\sim}75\;%$, respectively. Fifteen days of culture were required for the highest enzyme productivity. Among the sub-materials added, $30{\sim}40\;%$ of rice bran and $10{\sim}20\;%$ of defatted perilla seeds were effective for the enzyme production, but caused a reduced mycelial growth. The greatest effect of an addition of inorganic salts was obtained with $0.36{\sim}0.72\;%\;of\;(NH_4)_2HPO_4$. When 40 mesh or smaller rice straw and steam treatment at $0.5\;kg/cm^2$ were used, the mycelial growth decreased, whereas the enzyme production increased. The mycelial growth and enzyme production increased when $Ca(OH)_2$ was used as the alkali treatment, but decreased with increasing concentration of NaOH. As the fermentation proceeded, the amounts of ash, reducing sugar and total nitrogen increased, but cellulose, lignin and pentosan decreased. When the rice straw was treated with alkali, the amounts of ash, total nitrogen and lignin decreased, but reducing sugar and cellulose increased. At higher NaOH concentration, the variation become greater. The in vitro dry matter digestibility of the products increased from 55.03 % at the beginning of the fermentation to 62.72 % at 45 days after fermentation. The most effective alkali treatment on the digestibility of rice straw was KOH followed by NaOH. However, the digestibility increased with increasing concentration of NaOH. The digestibility of pretreated with alkali increased after fermentation as well.

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Disease Incidences in Radish and Chinese Cabbage. I. Major Diseases in Radish and Chinese Cabbage grown in Alpine Areas in Jeonbug Province (고냉지단경기채소(무우. 배추) 및 평야지 추작채소단지에 발생하는 주요병해조사 I. 전북고냉지단경기 무우, 배추 주요병해)

  • So I.Y.;Lee S.H.;Kim H.M.;Lee W.H.
    • Korean journal of applied entomology
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    • v.20 no.3 s.48
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    • pp.135-145
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    • 1981
  • Vegetables including Chinese cabbage and radish have been grown in alpine areas such as Muju, Namweon, Jinan and Jangsu during the shortage period of vegetables. The incidence of various diseases, environmental factors such as temperatures and rainfalls, and aphid populations as virus vector were observed at 10-day intervals from July to September in those areas. Disease incidence showed no significant difference among locations. Major diseases in Chin ese cabbage were bacterial soft rot, white spot, downy mildew, mosaic virus, Alternaria leaf spot and Fusarium seedling blight. Major diseases in radish were virus, white rust, black rot, root rot, leaf spot and Fusarium seedling blight. Disease incidence reached peak on Aug. 20 with $27\%$ infection in radish and with $20\%$ infection in Chinese cabbage during the growing season, and declined thereafter. Percentage of infection in each growth stage showed $25\%$ at root thickening stage and $26\%$ at the harvest time in radish; and the head formation stage, $24\%$ at the harvest time. The data indicate that disease incidence in radish increased rapidly at late growing stage and progressively increased in Chinese cabbage Seedling blight caused by Fusarium sp. and root rot caused by Aphanomyces sp. were also observed in those areas. Cool and wet weather appeared to be favorable for disease incidences during the rainy period of growing season although average temperature was about $25^{\circ}C$. Populations of aphids were lower in the alpine vegetable growing area than that of flat areas. Aphids as virus vectors from total aphids collected were $73.5\%$ or 289 virus vectors /993 total aphids in Namweon and $18.1\%$ or 31 virus vectors/171 total aphids in Muju. The most prerevalent species of aphids was Myzus persicae Sulz.

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Distribution of foodborne pathogens in red pepper and environment (고추와 재배환경의 식품매개 병원균 분포)

  • Jung, Jieun;Seo, Seung-Mi;Yang, SuIn;Jin, Hyeon-Suk;Jung, Kyu-Seok;Roh, Eunjung;Jeong, Myeong-In;Ryu, Jae-Gee;Ryu, Kyoung-Yul;Oh, Kwang Kyo
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.799-808
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    • 2021
  • This study was performed to investigate the extent of microbial contamination, the presence of enterotoxin genes, and the antibiotic susceptibility of Bacillus cereus in 58 red pepper plants and 43 environmental samples (soil, irrigation water, and gloves) associated with the plant cultivation. The detected counts of total aerobic bacteria, coliform bacteria, Escherichia coli, Bacillus cereus, and Staphylococcus aureus were lower in these samples, as compared to the regulations of standards for foods; moreover, pathogens, such as E. coli, E. coli O157:H7, Listeria monocytogenes, and Salmonella spp., were not detected. Genes encoding hemolysin BL enterotoxins (hblA, hblC, and hblD) as well as non-hemolytic enterotoxins (nheA, nheB, and nheC) were detected in 23 B. cereus specimens that were isolated from the test samples and had β-hemolytic activity. Interestingly, B. cereus is resistant to β-lactam and susceptible to non-β-lactam antibiotics. However, in this case, the isolated B. cereus specimens exhibited a shift from resistant to intermediate in response to cefotaxime and from susceptible to intermediate in case of rifampin, trimethoprim-sulfamethoxazole, vancomycin, clindamycin, and erythromycin. Therefore, the levels of B. cereus should be monitored to detect changes in antibiotic susceptibility and guarantee their safety.

Liquid-Composting Conditions of By-product Obtained from Degradation of Animal Carcass for Agriculture Recycling (폐가축사체의 농업적 재활용을 위한 가축사체 액상부산물의 액비화 조건 구명)

  • Seo, Young-Jin;Seo, Dong-Cheol;Kang, Se-Won;Lee, Sang-Gyu;Park, Ju-Wang;Choi, Ik-Won;Sung, Hwan-Hoo;Kang, Seog-Jin;Cho, Ju-Sik
    • Korean Journal of Environmental Agriculture
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    • v.32 no.4
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    • pp.348-354
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    • 2013
  • BACKGROUND: Globally, concern about emerging infectious diseases of livestock is growing. For the disposal of the animal carcass, it is necessary to recycle the carcass into an agriculturally usable product. The objective of this study was to investigate the composting conditions of liquid by-product obtained from degradation of animal carcass. METHODS AND RESULTS: Optimum conditions of liquid fertilizer were investigated using different microorganisms, pHs, and volumes of microorganisms (Lactobacillus rhamnosus+Pichia deserticola). Based on the results from the optimum conditions, compost maturity and quality of liquid fertilizer were evaluated for 112 days. The compost maturity of liquid fertilizer were higher in the order of LP(Lactobacillus rhamnosus + Pichia deserticola) > BC(Bacillus cereus) > BS(Bacillus subtilis). The optimum condition under different volumes of LP was injection of 0.5 mL/100 mL. The compost maturity under different pHs were higher in the order of pH 7 > $$5{\geq_-}9{\frac{._-}{.}}11$$. The liquid by-product at 56 days after composting was completely decomposed. The concentrations of T-N, T-P and $K_2O$ in liquid fertilizer at 56 days were 0.94, 0.17 and 3.78%, respectively, and the sum of those concentrations was 4.89%. CONCLUSION(S): Liquid fertilizer of by-product using pig carcass was decomposed with optimum conditions(LP, pH 7, injection of 0.5 mL/100 mL) in 56 days after composting, and was suitable for official standard of commercial fertilizer.