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Antibiotic Resistant Microbial Contamination (Enterobacter cloacae) Derived from Egg Yolk and Frozen Semen Extender in Porcine In Vitro Fertilized Embryos

  • Kwak, Seong-Song;Jeong, Se-Heon;Jang, Seung-Hoon;Jeon, Yu-Byeol;Nam, Young-Hee;Biswas, Dibyendu;Lee, Wan-Kyu;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.267-272
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    • 2010
  • The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

Cellular Responses and Morphological Changes of RDX-degrading Bacterium, Pseudomonas sp. HK-6 Exposed by Explosive Hexahydro-1,3,5-triaitro-1,3,5-triazine (RDX). (폭약 Hexahydro-1,3,5-trinitro-1,3,5-triazine(RDX)에 노출된 분해세균 Pseudomonas sp. HK-6의 세포반응과 형태변화)

  • 장효원;강형일;김치경;오계헌
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.75-82
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    • 2003
  • The cellular responses of RDX-degrading bacterium, Pseudomonas sp. HK-6 to explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) were examined. Strain HK-6 grown at different RDX concentrations was found to demonstrate the survival rate in proportional to the rate of the stress shock proteins produced in this bacterium. Analysis of total cellular fatty acid acids showed that lipids 10:0 iso and 14:1 $\omega$5c/$\omega$5t increased approx three times in strain HK-6 grown on RDX media than TSA media. SDS-PAGE and Western blot using anti-DnaK and GroEL revealed that several stress shock proteins including 70 kDa DnaK and 60 kDa CroEL were newly synthesized in strain HK-6 exposed to different RDX concentrations in exponentially growing cultures. 2-D PAGE of soluble protein fractions from the culture of HK-6 exposed to RDX demonstrated that approximately 300 spots were observed on the silver stained gel ranging from pH 3 to pH 10. As a result, 10 spots were significantly induced and expressed in response to RDX. Scanning electron microscopy fur the cells treated with 0.135 mM RDX for 12 hrs showed the presence of perforations and irregular rod shapes with wrinkled surfaces.

Autotrophic Perchlorate-Removal Using Zero-Valent Iron and Activated Sludge: Batch Test (영가철과 활성슬러지를 이용한 독립영양방식의 퍼클로레이트 제거: 회분배양연구)

  • Ahn, Yeong-Hee;Ha, Myoung-Gyu
    • Journal of Life Science
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    • v.21 no.3
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    • pp.444-450
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    • 2011
  • Perchlorate ($ClO_4^-$) is a contaminant found in surface water and soil/ground water. Autotrophic perchlorate-reducing bacteria (PRB) use hydrogen gas ($H_2$) as an electron donor to remove perchlorate. Since iron corrosion can produce $H_2$, feasibility of autotrophic perchlorate-removal using zero-valent iron (ZVI) was examined in this study using activated sludge that is easily available from a wastewater treatment plant. Batch test showed that activated sludge microorganisms could successfully degrade perchlorate in the presence of ZVI. The perchlorate biodegradation was confirmed by molar yield of $Cl^-$ as perchlorate was degraded. Scanning electron microscope revealed that rod-shaped microorganisms on the surface of iron particles used for the autotrophic perchlorate-removal, suggesting that iron particles could serve as supporting media for the formation of biofilm as well. DGGE analyses revealed that microbial profile of the inoculum (activated sludge) was different from that of biofilm sample obtained from the ZVI-added enrichment culture used for $ClO_4^-$-degradation. A major band of the biofilm sample was most closely related to the class Clostridia.

Studies on Proteolytic and Fibrinolytic Activity of Bacillus subtilis JM-3 Isolated from Anchovy Sauce (멸치액젓으로부터 분리한 Bacillus subtilis JM-3의 단백질 분해활성과 혈전 용해 활성에 관한 연구)

  • Lee, Sang-Soo;Kim, Sang-Moo;Park, Uk-Yeon;Kim, Hee-Yun;Shin, Il-Shik
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.283-289
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    • 2002
  • This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

Evaluation of Neutron Flux Distributions of SMART-P IST Region for the Design of Ex-Core Detector (SMART 연구로 노외계측기 설계를 위한 IST 영역의 중성자속 분포 평가)

  • Koo, Bon-Seung;Kim, Kyo-Youn;Lee, Chung-Chan;Zee, Sung-Quun
    • Journal of Radiation Protection and Research
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    • v.30 no.2
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    • pp.55-60
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    • 2005
  • The evaluation of neutron flux distribution was performed for the ex-core detector design of SMART-P. DORT and MCNP code were used for the calculation of energy-dependent neutron flux distribution at 100% full power condition. Two code results show that maximum thermal flux appears at the $1^{st}$ water region in IST region and agree within 10% difference. In addition, another evaluation was performed code with assumptions that cote was composed of fission source and control rod without fuel assemblies. These assumptions make neutron count rate to be minimized. As a results, maximum thermal flux showed $6.99{\times}10^{-2}(n/cm^2-sec)$, when the strength of initial fission source was assumed as $1.0{\times}10^8(n/sec)$. The main reason of these results is due to the thermalization of fast neutrons in the water region and thermal flux is proportional to 80% of total neutron flux. Therefore, optimization of filler material of detector guide tube, position of installation and axial length of detector segments is necessary for the design of ex-core detector to enhance the neutron count rate and above results could be used in ex-core detector design as a fluence requirement.

Exploring the Accuracy and Methods of Estimation on Base Physical Quantities (기본물리량 어림의 정확성 및 방법에 대한 탐색)

  • Song, Jin-Woong;Kim, Hae-Sun
    • Journal of The Korean Association For Science Education
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    • v.21 no.1
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    • pp.76-88
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    • 2001
  • This study explored people's accuracy and methods of estimating some base physical quantities, i.e. length, mass, time and temperature. A total of 40 members, ranging from freshmen to professors, of a physics education department of a local university were asked to make two different kinds of estimations, intuitive and operational, on two sets of objects. For intuitive estimation, they were asked to make estimations on four given objects (length - wood chopsticks, mass - rubber eraser, time electric fan, temperature - water in a cup) as soon as they faced with the objects, usually within a few seconds of seeing. For operational estimation, they were allowed to make estimations on a different set of objects (length - plastic rod, mass - lock, time - simple pendulum, temperature - water in a cup) with enough time and they could apply various available methods (e.g. using pencil to estimate the object's length, counting their own pulse rate to estimate time) for the estimation. The findings of this study can be summarized as follows: (1) for length, mass and temperature the intuitive estimations were better performed while for the time estimation the result was the reverse; (2) there was no positive relationship between the amount of physics experience and the accuracy of the estimation; (3) in general, people's accuracy of the length estimation was best performed while their mass estimation was worst performed; (4) people used their own various methods for estimation, esp. using nearby objects around them and applying mental units which have convenient values (e.g. 30cm, 50cm, 1kg, 1 Keun, 1 second).

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Fine Structure and Histopathological Changes Exposed to Acute High Salinity of the Gill of Japanese Clam, Corbicula japonica (일본재첩, Corbicula japonica 아가미의 정상구조와 고염분 급성노출에 의한 조직병리학적 변화)

  • Park, Jung-Jun;Lee, Jung-Sick;Lee, Jae-Seong
    • The Korean Journal of Malacology
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    • v.27 no.1
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    • pp.15-27
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    • 2011
  • This study was performed to observe ultrastructure of the gill and to ascertain the effect of salinity on histopathological and ultrastructural changes in the gill of the Japanese clam, Corbicula japonica. Experimental period was 7 days. Experimental groups consisted of control, 5, 10, 20 psu. $LC_{50}$ (96 h.) by the probit was 19.55 psu. Mortality was significantly different from the control (p < 0.05). Inner demibranch of the gill of C. japonica was wider 1.37 times than outer demibranch (p < 0.001). The filament zone on the plica can be distinguished by the six epithelial celll cell; frontal ciliated epithelium ($7{\mu}m$), latero-frontal ciliated epithelium ($5{\mu}m$), postlatero-frontal epithelim ($3{\times}8{\mu}m$), and lateral ciliated epithelium ($5{\mu}m$) in the frontal zone, endothelial cellin the intermediate zone, and abfrontal cell in the abfrontal zone. It had one type of secretory cell that was filled with fibrous substances of low electron density. The gill of C. japonica exposed to 5 psu for 7 days was observed partially disappearance of the cilia, and glycogen granule in the filament. In the 10 psu, gill appeared partially modification of epithelial cell and destruction of the glycocalyx. Gill exposed to 20 psu was extended nuclus of the ciliated epithelial cell, destruction of the organelles, and observed glycogen granules infiltration and numerous vacuoles. Moreover, more than 50% filaments were observed that come out chitinous rod from disappearance of epithelial cell in the filament. Therefore, the destruction of the cilia and epithelial cell induce physiological activity and it may be leading directly to death.

EFFECT OF RED GINSENG ON MICE EXPOSED TO VARIOUS STRESSES (홍삼이 스트레스에 노출된 생쥐의 행동에 미치는 영향)

  • Saito Hiroshi;Bao Tiantong
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.97-105
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    • 1984
  • Effect of water extract of red ginseng (Panax ginseng C.A. Meyer) from Nagano prefecture on (1) forced exercise in mice using rope climbing test, (2) extinction of memory in hanging stressed mice and rectal temperature during the exposure to hanging stress, (3) sex and learning behavior of chronic hanging stressed mice, (4) sex cycle in the adult female mice using chronic hanging stress, and (5) motor coordination and one trial passive avoidance response in $40\%$ alcohol administered mice using rotar-rod and step-through tests, were studied. Drugs tested were given orally. (1) When it was given before the forced exercise, it potentiated the performance of the forced exercise. When it was given just after the forced exercise once a day for 2 weeks, it protected the mice against the reduction of the performance on the forced exercise. (2) When it was given just after the stress once a day for 4 days, it delayed the extinction of passive avoidance response in both step through and stepdown tests, and protected the stressed mice against the decrease in rectal temperature slightly. (3) It protected the stressed mice against the decrease of sex behaviour and the increase in the failure of performance of passive avoidance response, and delayed the extinction of passive avoidanc

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Color and surface change of teeth restoration result of treatment of fruit extracts (과일추출물 처리에 따른 치아수복물의 색조 및 표면변화)

  • Choi, Eun Jung;Kim, Mi-Jin;Kim, Min-Ji;Park, Min-Ji;Shim, So-Yeon;Oh, Su-Jin;Lee, Ah-Min;Yang, Seok-Jun;Kang, Kyung-hee
    • Journal of Digital Convergence
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    • v.14 no.11
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    • pp.449-457
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    • 2016
  • At this study, we select five colorful and high-acid fruits: grape, lemon, orange, grapefruit and kiwi, and we made a composite resin specimen and dipped into fruit juice in order to figure out influence that affects on surface corrosion and coloring. We dipped resin specimen into fruit juice by three-day and seven-day interval at $37^{\circ}C$. Control group used the resin that was not dipped into fruit juice. For surface change according to the time, it was observed by using scanning electron microscopy, and it was measured by using spectrophotometer for change of coloring degree. As a result of surface change of resin by using scanning electron microscopy, furrow size between surface particles increased, and rectangular rod-shape structure was emerged, and small prominent particles on bumps disappeared. As a result of coloring degree of resin by measuring spectrophotometer, a difference between experimental group appears with the darkest brightness in the case where it is dipped into grape extracts.

Inhibition Effect of Ginseng Saponin on the Growth of Citrobacterer sp. Isolated from Contaminated Ginseng (오염된 인삼으로부터 분리된 Citrobacter sp.에 대한 인삼사포닌의 생육억제 효과)

  • Park, Chae-Kyu;Kwak, Yi-Seong;Hong, Soon-Gi;Lee, Hoon-Sang;Hwang, Mi-Sun;Rhee, Man-Hee;Won, Jun-Yeon;Han, Gyeong-Ho
    • Journal of Ginseng Research
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    • v.32 no.3
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    • pp.270-274
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    • 2008
  • A bacterium isolated from contaminated white ginseng was identified using API kit and electron microscope. This isolate was determined as rod shaped bacterium having about 1.0 ${\mu}m$ in diameter and 2.0 to 6.0 ${\mu}m$ in length. It had motility by peritrichous flagellum. The isolate had ${\beta}-galactosidase$, arginine dihydrolase and ornithin decarboxylase. It did not have ability not only to use citrate as sole carbon source and but also to produce $H_2S$. However, it could ferment glucose, manitol, sorbitol, rhamnose, arabinose and amygdalin. From these obserbations, the isolate was identified as Citrobacter sp. Ginseng saponin was added to culture of Citrobacter sp. in order to investigate saponin's influence on its growth. The strain was incubated at $38^{\circ}C$ for 3 days after addition of 0.05, 0.5, 2.0 and 4.0% (w/v) of saponin, respectively and the growth rates was investigated. The relative bacterial growth inhibition rates showed 28.6, 66.7, 92.4 and 97.7%, respectively, when compared with saponin non-treated group. These results suggest that the growth of Citrobacter sp. is inhibited by saponin in a concentration-dependent manner.