• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.37-37
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• 2015

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

• Applied Microscopy
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• v.40 no.2
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• pp.89-99
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• 2010

Fibroblasts are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. Mitochondria produce ATP through oxidative metabolism to provide energy to the cell under physiological conditions. Also, mitochondrial dysfunction and oxidative stress have been implicated in cellular senescence and aging. Alternations in mitochondrial structure and function are early events of programmed cell death or apoptosis and mitochondria appear to be a central regulator of apoptosis in most somatic cell. Clonorchis sinensis, one of the most important parasite of the human bile duct in East Asia, arouses epithelial hyperplasia and ductal fibrosis. Isolated fibroblast from the bile ducts of rats infected by C. sinensis showed increase of cytoplasmic process. In addition, decrease of cellular proliferation was observed in fibroblasts which was isolated from normal rat bile duct and then cultured in media containing C. sinensis excretory-secretory product. However, the effects of C. sinensis infection on the mitochondrial enzyme distribution is not clearly reported yet. Therefore, we investigated the structural change of C. sinensis infected bile duct and mitochondrial enzyme distribution of the cultured fibroblast isolated from the C. sinensis infected rat bile duct. As a result, C. sinensis infected SD rat bile ducts showed the features of chronic clonorchiasis, such as ductal connective and epithelial tissue dilatation, or ductal fibrosis. In addition, fibroblast in ductal connective tissue was damaged by physical effect of fibrotic tissue and chemical stimulation. Immunohistochemically detected mitochondrial electron transferase (ATPase, COXII, Porin) was decreased in C. sinensis infected rat bile duct and cultured fibroblast from infected rat bile duct. It can be hypothesized that the reason why number of electron transferase decrease in fibroblast isolated from the rat bile duct infected with C. sinensis is because dysfunction of electron transport system is occurred mitochondrial dysfunction, increase of ROS (reactive oxygen species) and apoptosis after chemical damage on the cell caused by C. sinensis infection. Overall, C. sinensis infection induces fibrotic change of ductal connective tissue, mutation of cellular metabolism in fibroblast and mitochondrial dysfunction. Consequently, ductal fibrosis inhibits fibroblast proliferation and decreases mitochondrial electron transferase on fibroblast cytoplasm. It was assumed that the structure of bile duct could not normalized and ductal fibrosis was maintained for a long period of time according to fibroblast metamorphosis and death induced by mitochondrial dysfunction.

• Journal of Preventive Medicine and Public Health
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• v.3 no.1
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• pp.1-16
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• 1970

The present status of control measures for public health important helminthic infections in Korea was surveyed in 1969 and the following results were obtained. The activities of parasitic examination and Ascaris treatment for the positives which were done during 1966 to 1969 were brought in poor result and could not decrease the infection rate. It is needed to improve or strengthen the activities. The mass treatment activities for paragonimiasis and clonorchiasis in the areas which were designated by the Ministry of Health were carried out during 1965 to 1968 with no good results in decrease of estimated number of the patients. There were too many pharmaceutical companies where many kinds of anthelmintics were produced. It may be better to reduce the number of anthelmintics produced and control the quality. The human feces, the most important source of helminthic infections, was generally not treated in sanitary ways because of the poor sewerage system and no sewage treatment plant in urban areas and insanitary latrines in rural areas. The field soils of 170 specimens were collected from 34 areas out of 55 urban and tourist areas where night soil has been prohibited by a regulation to be used as a fertilizer, and examined for parasites contamination with the result of Ascaris egg detection in 44%. Some kinds of vegetables of 64 specimens each from the supply agents of parasite free vegetables and general markets were collected and examined for parasites contamination with the results of Ascaris egg detection in 25% and 36% respectively. The parasite control activities and the ability of parasitological examination techniques in the health centers of the country were not satisfactory. The budget of the Ministry of Health for the parasite control was very poor. The actual expenditure needed for cellophane thick smear technique was 8 Won per a specimen. As a principle the control of helminthic infections might be led toward breaking the chain of events in the life cycle of the prasites and eliminating environmental and host factors concerned with the infections, and the following methods nay be pointed out. 1) Mass treatment might be done to eliminate human reservoirs of an infection. 2) Animal reservoirs which are related with human infections night be eliminated. 3) The excretes of reservoirs, particularly human feces, should be treated in sanitary ways by the means of sanitary sewerage system and sewage treatment plant in urban areas and sanitary latrines such as waterborne latrine, aqua privy and pit latrine in rural areas. The increase of national economical development and prohibition of the habit of using night soils as a fertilizer might be very important factors to achieve the purpose. 4) The control of vehicles and intermediate hosts might be done by the means of prohibition of soil contamination with parasites, food sanitation, insect control and snail control. 5) The improvement of insanitary attitudes and bad habits which are related with parasitic infections night be done by the means of prohibition of habit of using night soils as a fertilizer, and improving eating habits and personal hygiene. 6) Chemoprophylactic measure and vaccination may be effective to prevent the infections or the development of a parasite to adult in the bodies when the bodies were invaded by parasites. Further studies and development of this kind of measures are needed.

• Food Science and Preservation
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• v.14 no.1
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• pp.35-41
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• 2007

As the consumption of environmentally friendly agricultural products increases, food safety is at the forefront of public health concerns. We analyzed the biological hazards of 26 species of leaf vegetables and 4 species of fruit according to types of cultivation (conventional or organic filming) and distribution system (giant retailers or organic food stores) using various culture media, automatic bacterial identification systems, and microscopy, Total bacterial count of unwashed agricultural product ranged from $5.2{\times}10^{3}\;to\;1.5{\times}10^{5}\;CFU/mL$ (from 0.1 g of agricultural products), and the average count dropped 25-fold (range, 8-60-fold) after water washing. Microbial levels of washed organic agricultural products were $6.0{\times}10^{2}-1.9{\times}10^{4}\;CFU/mL$, and were not significantly different f개m the microbial loads on conventionally farmed produce. There was no significant difference in bacterial count from agricultural produce purchased from giant retailers or organic food stores. Total microbial count of Chinese cabbage, welsh onion, red chicory and kale were comparatively high, and Enterobacter cloacae was isolated most frequently. Parasites were detected in agricultural products purchased from conventional farm products in the stores of giant retailers, and in organic food stores, and parasite prevalence was especially high in Chinese cabbages and welsh onion. The study indicated that cultivation methods and distribution systems did not cause significant differences in biological contamination levels of agricultural produce. Some vegetables and fruits were highly contaminated effective sanitizing methods to reduce these biological hazards are needed.

• Parasites, Hosts and Diseases
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• v.26 no.1
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• pp.39-44
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• 1988

A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.

• Journal of Food Hygiene and Safety
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• v.24 no.2
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• pp.174-181
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• 2009

The consumption of "ready-to-eat" agricultural products is recently increasing and the safety of these agricultural products is forefront of public concerns. The 120 samples of paprikas, strawberries and tomatoes, which are the representative exported agricultural products, were purchased at the department stores and discount stores in Daejeon. And we determined the microbiological and parasitological contamination level of these agricultural products using culture media, multiplex PCR, commercial bacterial detection kit and microscopy, and also evaluated the decontamination method. Mean counts of total aerobic bacteria from these agricultural products ranged from $1.3{\times}10^4$ CFU/g to $1.8{\times}10^5$ CFU/g, and mean counts of coliforms ranged from $1.4{\times}10^3$ CFU/g to $9.6{\times}10^3$ CFU/g. There was no significant difference in the level of bacterial contamination between the agricultural products from department stores and the ones from discount stores. Strawberry showed the highest contamination level for the bacteria and we also found the unidentified parasite eggs. Enterobacter cloacae was the most frequently isolated bacteria strain, but no food poisoning pathogenic bacteria except Staphylococcus aureus was isolated from the products by multiplex PCR. Compared to unwashed products, tab water-washed ones showed 80% decrease of the counts of total aerobic bacteria on the agricultural products, and the rates decreased more by incorporating detergent or ultrasonic wave treatment. We concluded that the biological contamination levels among paprikas, strawberries and tomatoes were the highest in strawberries, but there were not significant difference according to distribution systems.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.720-728
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• 2001

Cercaria yamagutii Ito, 1957 (C. yamagutii) was found in Lunatia fortuni (L. fortuni) and Neverita didyma (N. didyma) collected from the tideland of Sim-po located at the estuary of the Mankyong River, Chonbuk. It was finally confirmed that the parasite is Acanthoparyphium tyosenense Yamaguti, 1939 (A. tyosenense) and its life history was clarified in this study. Mactra veneriformis (M. veneriformis) was artificially infected with C. yamagutii isolated from L. fortuni and N. didyma. It began to intrude into M. veneriformis through the inhalent canal. Five hours after infection, the tails of the cercaria began to be separated from the main body and the cercaria started to form cysts. Mature cysts were formed 340 hours (14 days) after infection. The cysts were $300\sim360{\mu}m$ in diameter and the encysted metacercarias were $790\sim800\times300\sim310{\mu}m$ in size. The metacercarias were administered orally to Larus crassiostris (L. crassiostris), and adult worms of $84.5\sim112.5\times55\sim65{\mu}m$ were found full of eggs with $2.20\~3.70$ mm long and $0.40\~0.59$ mm wide after 10 days. In a field study, it was observed that the infection rate of A. tyosenense is $99.5\%$ in M. venerifomis, $76.3\%$ Solen strictus (S. strictus), and $37\%$ Ruditapes philippinarum (R. philippinarum), No difference was found among different host sizes, It was concluded that the first intermediate hosts of A. tyosenense Yamaguti were L. fortuni, N. didyma, Tympanotonus microptera, Cerithidea (Cerithidea) largillierti, Cerithidea (Cerideopsilloa) cingulata, the second intermediate hosts M. venerifomis, S. strictus and R. philippinamn, and the final hosts L. crassiostris and Melanitta fusca stejnegeri.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.17 no.4
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• pp.292-302
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• 2012

This paper reviews comparison analysis of current and latest application for stock identification methods of Todarodes pacificus, and the pros and cons of each method and consideration of how to compensate for each other. Todarodes pacificus which migrates wide areas in western North Pacific is important fishery resource ecologically and commercially. Todarodes pacificus is also considered as 'biological indicator' of ocean environmental changes. And changes in its short and long term catch and distribution area occur along with environmental changes. For example, while the catch of pollack, a cold water fish, has dramatically decreased until today after the climate regime shift in 1987/1988, the catch of Todarodes pacificus has been dramatically increased. Regarding the decrease in pollack catch, overfishing and climate changes were considered as the main causes, but there has been no definite reason until today. One of the reasons why there is no definite answer is related with no proper analysis about ecological and environmental aspects based on stock identification. Subpopulation is a group sharing the same gene pool through sexual reproduction process within limited boundaries having similar ecological characteristics. Each individual with same stock might be affected by different environment in temporal and spatial during the process of spawning, recruitment and then reproduction. Thereby, accurate stock analysis about the species can play an efficient alternative to comply with effective resource management and rapid changes. Four main stock analysis were applied to Todarodes pacificus: Morphologic Method, Ecological Method, Tagging Method, Genetic Method. Ecological method is studies for analysis of differences in spawning grounds by analysing the individual ecological change, distribution, migration status, parasitic state of parasite, kinds of parasite and parasite infection rate etc. Currently the method has been studying lively can identify the group in the similar environment. However It is difficult to know to identify the same genetic group in each other. Tagging Method is direct method. It can analyse cohort's migration, distribution and location of spawning, but it is very difficult to recapture tagged squids and hard to tag juveniles. Genetic method, which is for useful fishery resource stock analysis has provided the basic information regarding resource management study. Genetic method for stock analysis is determined according to markers' sensitivity and need to select high multiform of genetic markers. For stock identification, isozyme multiform has been used for genetic markers. Recently there is increase in use of makers with high range variability among DNA sequencing like mitochondria, microsatellite. Even the current morphologic method, tagging method and ecological method played important rolls through finding Todarodes pacificus' life cycle, migration route and changes in spawning grounds, it is still difficult to analyze the stock of Todarodes pacificus as those are distributed in difference seas. Lately, by taking advantages of each stock analysis method, more complicated method is being applied. If based on such analysis and genetic method for improvement are played, there will be much advance in management system for the resource fluctuation of Todarodes pacificus.

• Journal of agricultural medicine and community health
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• v.1 no.1
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• pp.10-21
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• 1976

The traditional application of night soil to vegetable gardens and rice paddies results in a most wide spread condition of parasitism, with a variety of helminths found in Korea. In addition to the above fact, the peculiar habit of the consumption of raw vegetables, fish, crustaceans and mammals provides a means of infestations of helminths. During the last sixty years numerous reports were found on the prevalence of helminths amongst the Korean population in different parts of the country, and it was generally recognized that ascariasis, hookworm disease, filariasis, clonorchiasis and paragonimiasis constitute the important helminthic disease in Korea. In practical measures of parasite control activities the main measures are summarized as mass-treatment, night-soil disposal and transmission control. Among the three, the mass-treatment has been commonly applied, however, no reduction of transmission has been obtained by treatment of a population. Therefore, the ultimate eradication of parasites will depend upon the application of comprehensive environmental sanitation measures. The basic environmental measures will be concerned with (a) the safe disposal of human excreta, (b) the provision of adequate and safe water supplies in such a way as to promote a higher standard of personal hygiene in the population, and (c) the prevention of food contamination by faecal material. Additional environmental measures will deal with the improvement of housing and housing hygiene and with general community development. Community development means social and cultural as well as economic development. The control measures on the parasitic endemic diseases, such as clonorchiasis and paragonimiasis are the good examples for community health development in Korea. The control of Clonorchis and Paragonimus infections are theoretically very simple, as the infection can only invade the human body by way of encysted metacercaria which are taken into the body when eating passive intermediate hosts(fishes, crabs and crayfishes). Although prophylactic measures in the case of the infections deal with above merely consist the fishes in cooking or submerging in hot water before eating them, it is exceedingly difficult to carry out such simple measures in face of century old traditions, to which the relatively primitive population clings with great tenacity. There is no one universally applicable method of control. The choice of methods must be dictated by the nature of the environment. the habit and custom of the people. the pattern of transmission and the resources of the country. There must exist a well organized public health infrastructure. Since a control programme is of necessity on a longterm basis and continuity in its implementation is essential. An investigation should be made on the prevalence of the diseases and its relationships to irrigation engineering, freshwater ecology, agricultural methods, hydro-electric schemes, and the development of communities in affected areas. In conclusion, however. the control of clonorchiasis and paragonimiasis in Korea is not an impossible task. A combination of efforts with major emphasis on health education and mass chemotherapy coupled with governmental aid in enforcing legislative public health measures could reduce the diseases. Health education in particular attempts following four things: (a) It supplies a person with enough general knowledge about a disease to make the preventive measures. (b) It makes a person feel sufficiently about the importance of his own health to make him alter his behavior and adopt these preventive measure. (c) It makes him concerned for the health others. (d) It tries to make him feel so strongly about the first three that be supports and even initiates preventive action by the community. Educational efforts should be directed primarily toward school children because it is during the early years that most persons become infected, and also because children are less entrenched in their food habits so that, the educational process should be involved at various levels in successive changes of knowledge, attitude, beha viour, habit and custom of their lives. The most parasitic endemic diseases are related to community diseases. In caring for a sick community. the first stage is to gather epidemiological data, the next is to make inferences from it-to make the community diagnosis. The third is to prescribe community treatment or community health action part of a community health action programme. The community health action is the sum of the steps decided upon to remedy the critical features revealed by the community diagnosis. Action takes various forms; health education is the most important.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.