CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))
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- Korean Journal of Veterinary Research
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- v.9 no.1
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- pp.23-62
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- 1969
Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about
The detail survey on the Songsanri tomb site including the Muryong royal tomb was carried out during the period from May 1 , 1996 to April 30, 1997. A quantitative analysis was tried to find changes of tomb itself since the excavation. Main subjects of the survey are to find out the cause of infiltration of rain water and groundwater into the tomb and the tomb site, monitoring of the movement of tomb structure and safety, removal method of the algae inside the tomb, and air controlling system to solve high humidity condition and dew inside the tomb. For these purposes, detail survery inside and outside the tombs using a electronic distance meter and small airplane, monitoring of temperature and humidity, geophysical exploration including electrical resistivity, geomagnetic, gravity and georadar methods, drilling, measurement of physical and chemical properties of drill core and measurement of groundwater permeability were conducted. We found that the center of the subsurface tomb and the center of soil mound on ground are different 4.5 meter and 5 meter for the 5th tomb and 7th tomb, respectively. The fact has caused unequal stress on the tomb structure. In the 7th tomb (the Muryong royal tomb), 435 bricks were broken out of 6025 bricks in 1972, but 1072 bricks are broken in 1996. The break rate has been increased about 250% for just 24 years. The break rate increased about 290% in the 6th tomb. The situation in 1996 is the result for just 24 years while the situation in 1972 was the result for about 1450 years. Status of breaking of bircks represents that a severe problem is undergoing. The eastern wall of the Muryong royal tomb is moving toward inside the tomb with the rate of 2.95 mm/myr in rainy season and 1.52 mm/myr in dry season. The frontal wall shows biggest movement in the 7th tomb having a rate of 2.05 mm/myr toward the passage way. The 6th tomb shows biggest movement among the three tombs having the rate of 7.44mm/myr and 3.61mm/myr toward east for the high break rate of bricks in the 6th tomb. Georadar section of the shallow soil layer represents several faults in the top soil layer of the 5th tomb and 7th tomb. Raninwater flew through faults tnto the tomb and nearby ground and high water content in nearby ground resulted in low resistance and high humidity inside tombs. High humidity inside tomb made a good condition for algae living with high temperature and moderate light source. The 6th tomb is most severe situation and the 7th tomb is the second in terms of algae living. Artificial change of the tomb environment since the excavation, infiltration of rain water and groundwater into the tombsite and bad drainage system had resulted in dangerous status for the tomb structure. Main cause for many problems including breaking of bricks, movement of tomb walls and algae living is infiltration of rainwater and groundwater into the tomb site. Therefore, protection of the tomb site from high water content should be carried out at first. Waterproofing method includes a cover system over the tomvsith using geotextile, clay layer and geomembrane and a deep trench which is 2 meter down to the base of the 5th tomb at the north of the tomv site. Decrease and balancing of soil weight above the tomb are also needed for the sfety of tomb structures. For the algae living inside tombs, we recommend to spray K101 which developed in this study on the surface of wall and then, exposure to ultraviolet light sources for 24 hours. Air controlling system should be changed to a constant temperature and humidity system for the 6th tomb and the 7th tomb. It seems to much better to place the system at frontal room and to ciculate cold air inside tombs to solve dew problem. Above mentioned preservation methods are suggested to give least changes to tomb site and to solve the most fundmental problems. Repairing should be planned in order and some special cares are needed for the safety of tombs in reparing work. Finally, a monitoring system measuring tilting of tomb walls, water content, groundwater level, temperature and humidity is required to monitor and to evaluate the repairing work.