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An Outbreak of Epidemic Keratoconjunctivitis by Adenovirus Type 8 in a Neonatal Intensive Care Unit (신생아 중환자실에서의 아데노바이러스 8형에 의한 유행성 각결막염의 발생)

  • Park, Na-Ri-Mi;Na, Ji-Youn;Joung, Kyoung-Eun;Lee, Ji-Na;Kim, Ee-Kyung;Kim, Han-Suk;Kim, Seong-Joon;Song, Jung-Suk;Oh, Hyang-Soon;Lee, Hoan-Jong;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.15 no.1
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    • pp.44-53
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    • 2008
  • Purpose : Epidemic keratoconjunctivitis (EKC) caused by adenovirus is a highly contagious disease, which has been reported as outbreaks involving adults in the community. However, there has been no report on EKC outbreak by adenovirus in a neonatal intensive care unit (NICU) in Korea. Aims of this study were to investigate the EKC outbreak by adenovirus type 8 in NICU and to confirm an effectiveness of polymerase chain reaction (PCR) for diagnosis. Methods : Conjunctival swab or nasopharyngeal aspirate specimens were taken from all patients and tested by viral culture and PCR. Adenovirus serotype was determined by sequencing of PCR product of selected region of hexon gene using the virus isolates or specimens. Results : An outbreak of EKC occurred which was involving 12 preterm infants in the NICU of the Seoul National University Children's Hospital between July 12th and August 1st, 2005. Three hospital staffs and one family member of the neonate were also affected. Adenovirus was detected in 12/12 (100%), 6/11 (54.5%) by PCR and virus culture, respectively. Eleven PCR-positive neonates were identified as serotype 8 by sequencing. The first affected 4 babies have had routine ROP (retinopathy of prematurity) examinations one week ago. While previous outbreaks were sustained for a few months, the event in our unit was controlled without complications in 3 weeks. Conclusion : We analyzed the EKC outbreak by adenovirus type 8 in NICU. Adenovirus serotype was identified by PCR and sequencing with high sensitivity for the first time in Korea, so we suggest this method can be very useful for rapid diagnosis and infection control.

Genomic analysis of Mycobacterium fortuitum by pulsed-field gel electrophoresis (Pulsed-field Gel Electrophoresis를 이용한 Mycobacterium fortuitum의 유전형 분석)

  • Lee, Tae-Yoon;Do, In-A;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.366-385
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    • 1995
  • Epidemiological studies are important in both the prevention and treatment of mycobacterial infections. This study was initiated to establish the pulsed-field gel electrophoresis (PFGE) method, which are not yet extensively studied. The most apprpriate restriction endonucleases included DraI, AsnI, and XbaI. The optimal PFGE condition was different according to the enzymes used. Two stage PFGE was performed, in case of DraI first stage was performed with 10 seconds of initial pulse and 15 seconds of final pulse, while the second stage was performed with 60 seconds of initial pulse and 70 seconds of final pulse. The electrophoresis time for DraI-PFGE was 14 hours for each stage. Electrophoresis was performed for 22 hours, in case of XbaI, with 3 seconds of initial pulse and 12 seconds of final pulse. Electrophoresis was performed for 22 hours, in case of AsnI, with 5 seconds of initial pulse and 25 seconds of final pulse. In all cases the voltage of the electrophoresis was maintained constantly at 200 voltage. Standard mycobacterial strains, which included Mycobacterium bovis BCG, M. tuberculosis, and M. fortuitum, could not be differentiated by PFGE analysis. PFGE analysis was performed to differentiate 9 clinically isolated M. fortuitum strains using AsnI. All M. fortuitum strains showed different genotypes except 2 strains. Cluster analysis divided M. fortuitum strains into 2 large groups. PFGE analysis was performed to further differentiate M. fortuitum isolates using XbaI. The undifferentiated 2 M. fortuitum strains showed different PFGE patterns with Xba I. Cluster analysis of the XbaI-PFGE patterns showed more complex grouping than AsnI-PFGE patterns, which showed that XbaI-PFGE analysis was better than AsnI-PFGE in M. fortuitum genotyping. The top dissimilarity values of AsnI-PFGE and XbaI-PFGE were 0.74 and 0.75, respectively. This value was higher than that of arbitrarily primed polymerase chain reaction (AP-PCR) analysis and lower than that of restriction fragment length polymorphism (RFLP) analysis. This suggested that PFGE can be used as a supportive or alternative genotyping method to RFLP analysis.

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A New Short Growth-Duration Rice Cultivar, "Keumo 3" (소득작물 전후작용 단기성 벼 품종 "금오3호")

  • Kang, Jong-Rae;Lee, Jong-Hee;Kwack, Do-Yeon;Lee, Jeom-Sik;Park, No-Bong;Ha, Woon-Gu;Park, Dong-Soo;Yeo, Un-Sang;Lim, Sang-Jong;Oh, Byeong-Geun
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.292-298
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    • 2009
  • A new rice cultivar "Keumo 3" was developed for adopting under double cropping system with after or before cash crop cultivation. It was selected from the cross-combination between YR17202 $F_2$/Shinkeumobyeo//YR15727-B-B-B-102. The parent, YR17202 $F_2$ individual plant, was used for tolerance to lodging, it derived from a cross between Nonganbyeo/Shinkeumobyeo. Nonganbyeo is well known to lodging tolerance cultivar, as well as biotic stress, because it was developed by crossing with Tongil type. And the YR15727-B-B-B-102 line used as another parent with short growth duration, likewise highly resistance to rice blast disease. The pedigree derived from the cross-combination YR17202 $F_2$/Shinkeumobyeo//YR15727-B-B-B-102 were generated to $F_7$, and a best line among them named as Milyang 201. After a series of yield trials, including local adaptability test conducted throughout the peninsular of Korea, Milyang 201 was registered with the name of "Keumo 3" in 2005. The cultivar belongs to a early maturing group and heads 4 days earlier than Keumobyeo, a standard cultivar. It has short culm, and less spikelet number per panicle than Keumobyeo. However, its milled rice yield grown under extremely late transplanting time, 10. July, over the 3 local sites for 2003-2005 years, averaged 4,48 MT/ha, which is 6% higher than the standard, Keumobyeo. "Keumo 3" has showed a durable resistance to leaf blast disease during fourteen blast nurseries screening covered from south to north in Korea for 2003-2007 years. And it was confirmed harbours pi-zt, a durable blast resistance gene. Moreover it was incompatible with 19 blast isolates under artificial inoculation, except one isolate, K1101. Additionally, "Keumo 3" exhibits resistance to $K_1$, $K_2$ and $K_3$ of bacterial blight pathogen, as well as strip virus disease resistance, and moderate resistance to dwarf virus disease. Consequently, the new rice cultivar "Keumo 3" would be well adopted where a bio stress makes a big problem annually.

Abundance of Harmful Algae, Cochlodinium polykrikoides, Gyrodinium impudicum and Gymnodinium catenatum in the Coastal Area of South Sea of Korea and Their Effects of Temperature, Salinity, Irradiance and Nutrient on the Growth in Culture (남해안 연안에서 적조생물, Cochlodinium polykikoides, Gyrodinium impudicum, Gymnodinium catenatum의 출현상황과 온도, 염분, 조도 및 영양염류에 따른 성장특성)

  • LEE Chang Kyu;KIM Hyung Chul;LEE Sam-Geun;JUNG Chang Su;KIM Hak Gyoon;LIM Wol Ae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.536-544
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    • 2001
  • Three harmful algal bloom species with similar morphology, Cochlodinium polykrikoides, Gyodinium impudicum and Gymodinium catenatum have damaged to aquatic animals or human health by either making massive blooms or intoxication of shellfishes in a food chain. Eco-physiological and hydrodynamic studies on the harmful algae offer useful informations in the understanding their bloom mechanism by giving promising data for the prediction and modelling of harmful algal blooms event. Thus, we studied the abundance of these species in the coastal area of South Sea of Korea and their effects of temperature, salinity, irradiance and nutrient on the growth for the isolates. The timing for initial appearance of the three species around the coastal area of Namhaedo, Narodo and Wando was between Bate July and late August in 1999 when water temperature ranged from $22.8^{\circ}C\;to\;26.5^{\circ}C$ Vegetative cells of C. polykrikoides and G. impudicum were abundant until late September when water temperature had been dropped to less than $23^{\circ}C$. By contrast, vegetative cell of G. catenatum disappeared before early September, showing shorter period of abundance than the other two species in the South Sea. Both G. impudicum and G. catenatum revealed comparatively low density with a maximal cell density of 3,460 cells/L and 440 cells/L, respectively without making any bloom, while C. polykrikoides made massive blooms with a maximal cell density more than $40\times10^6$cells/L, The three species showed a better growth at the relatively higher water temperature ranging from 22 to $28^{\circ}C$ with their maximal growth rate at $25^{\circ}C$ in culture, which almost corresponded with the water temperature during the outbreak of C. polykrikoides in the coastal area of South Sea. Also, they all showed a relatively higher growth at the salinity from 30 to $35\%$. Specially, G. impudicum showed the euryhalic characteristics among the species, On the other hand, growth rate of G. catenatum decreased sharply with the increase of water temperature at the experimental ranges more than $35\%$. The higher of light intensities showed the better growth rates for the three species, Moreover, C. polykrikoides and G. impudirum continued their exponential growth even at 7,500 lux, the highest level of light intensity in the experiment, Therefore, It is assumed that C. polykrikoides has a physiological capability to adapt and utilize higher irradiance resulting in the higher growth rate without any photo inhibition response at the sea surface where there is usually strong irradiance during its blooming season. Although C. poiykikoides and G. impudicum continued their linear growth with the increase of nitrate ($NO_3^-$) and ammonium ($NH_4^-$) concentrations at less than the $40{\mu}M$, they didn't show any significant differences in growth rates with the increase of nitrate and ammonium concentrations at more than $40{\mu}M$, signifying that the nitrogen critical point for the growth of the two species stands between 13.5 and $40{\mu}M$. Also, even though both of the two species continued their linear growth with the increase of phosphate ($PO_4^{2-}$) concentrations at less than the $4.05{\mu}M$, there were no any significant differences in growth rates with the increase of phosphate concentrations at more than $4.05{\mu}M$, signifying that the phosphate critical point for the growth of the two species stands between 1.35 and $4.05{\mu}M$. On the other hand, C. polykrikoides has made blooms at the oligotrophic environment near Narodo and Namhaedo where the concentration of DIN and DIP are less than 1.2 and $0.3{\mu}M$, respectively. We attributed this phenomenon to its own ecological characteristics of diel vertical migration through which C. polykrikoides could uptake enough nutrients from the deep sea water near bottom during the night time irrespective of the lower nutrient pools in the surface water.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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