• Parasites, Hosts and Diseases
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• v.25 no.2
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• pp.159-167
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• 1987

To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10~15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10kDa bands. Two bands in sparganum extract (130 and 64kDa) and two bands in hydatid cyst fluid (52 and 27kDa) were cross-reacting bands with sera from cysticercosis patients. Saline extracts of Fasciola, ClonorchiJ and Paragonimus did 'not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7kDa.

• Parasites, Hosts and Diseases
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• v.23 no.1
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• pp.95-101
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• 1985

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.171-176
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• 1986

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay (ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: 1. The specificity (95% confidence interval in negative sera of bovine fascioliasis; Mean+$2{\times}SD$ of absorbence) of the first (MW>150,000) and the second antigens(MW 120,000) were 93.7%, but those of others including crude antigen showed 100%. 2. The sensitivity(positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6%, 87.5% and 0%, respectively, but those of others showed all 100%. 3. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8. 43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. 4. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the fourth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. 5. The wheal size for bovine infected with F. hepatica was $2.46{\pm}0.15cm$ in the intradermal test antigen(saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that. of intradermal test antigen, whereas those of the fouth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

• Korean Journal of Veterinary Research
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• v.14 no.1
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• pp.59-71
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• 1974

The studies were conducted on the allergic skin reaction of bovine eurytremiasis. In order to confirm the appearance of allergic skin reaction in this disease, a protein fraction of Eurytrema SPP(EPF) was extracted, and it's antigenicity was studied by means of intradermal reaction on the rabbits, goats and bovine. Rabbits used in this studies were divided into three groups: the first group was sensitized with EPF, the second group was sensitized with EPF, and the third group was served as control which is consisted with 5 non-sensitized and 2 saline injected rabbits. Three healthy native goats and bovine also used. Among these, 86 cases were eurytremiasis, 31 cases were fascioliasis and 71 cases non-infected. A protein fraction was also made from adult worm of Fasciola SPP(FPF) and tested the cross reaction between these two different fluke infestations. The results obtained were as follow: 1. Intradermal reaction was appeared in the infected cattle(bovine eurytremiasis) as well as in the EPF sensitized rabbits and goats. 2. In the EPF sensitized rabbits, the wheal of the intradermal reaction reacted the maximum size (increased wheal size of 5.0 to 9.0 mm) in 40 to 60 minutes and erythema in 60 minutes after intradermal injection of EPF antigen. In the EPF sensitized goats, the size of wheal reacted the maximum(increased wheal size of 5.5 to 8.0 mm) in 30 to 40 minutes, and then disappeared in 120 to 180 minutes after injection of EPF antgen. On the contrary, in the control groups of rabbits and goats, the wheal (7.0 mm) produced following antigen injection reached only to 7.5 to 8.5 mm. in 10 minutes without developing erythema, and then perished in 40 to 60 minutes after injection of EPF antigen. In the infected cattle, the wheal of the intradermal reaction reached the maximal sizes(increased wheal size of 4.0 to 11.0 mm) in 15 to 20 minutes and perished in 60 to 90 minutes after injection of EPF antigen. 3. It would be standardized that the positive reaction is above 4.0 mm, the negative below 2.0 mm and the suspective is 3.0 mm of the infected cattle. In the results of intradermal reaction, 87.2%(75 of 86 cases) was positive, 4.7%(4 of 86 cases) negative and 8.1% (7 of 86 cases) was suspective. However, in the non-infected control cattle, the wheal produced following injection disappeared in 30 to 40 minutes without increasing the wheal size in the most subjects or with incresing only to 1.0 to 2.0 mm in a few cattle. 4. In the results of intradermal reaction which were reciprocally tested in EPF sensitized rabbits by FPF antigen and in FPF sensitized. rabbits by EPF antigen, there were cross reacted. In the 31 cases if fascioliasis tested by EPF antigen, 11 cases were not specifically reacted with EPF antigen. Judging from the results of cross reaction in experimental rabbits and natural bovine fascioliasis, it would be expected when both EPF and EPF antigens are given intracutaneously on the same bovine patients of eurytremiasis or fascioliasis, the former antigen will produce the larger wheal in eurytremiasis, while the latter antigen will produce also the larger wheal in fascioliasis.

• Journal of The Korean Society of Grassland and Forage Science
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• v.3 no.2
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• pp.100-111
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• 1983

The research results reported herein had the objectives to understand and analyze the present problems of saemaeul woodland joint cattle grazing system in Kang Won Do and to take steps of improvement. The study results on actual management conditions, problems analyzed and improvement plan of total 208 joint cattle grazing area which was established 105 area in 1981 and 103 area in 1982 were summarized as follows: 1. the effectiveness of joint cattle grazing projects 1) Average daily weight gain of cattle during joint cattle grazing period was 0.4kg, showing higher daily than the conventional feeding of 0.33kg. 2) Increase of total farm income over the conventional feeding system were \1,031,357,320 during the grazing period from May to October in 1982 by adapting the 208 joint cattle grazing system, of which effectiveness of weight gain was \293,075,300 and labor saving was \543,838,750. 3) According to the results of questionaire investigation from 208 joint cattle grazing area, effectiveness of joint cattle grazing system over the conventional system were (1) labor saving, (2) feed cost saving, (3) reduced diseases, (4) increase of number of feeding, (5) inspiration of joint endeavor, (6) effect of more gain, (7) easiness of feeding and feed cost savings. 2. Problems of joint cattle grazing system. 1) Shortages of grass were a problem at second year of joint cattle grazing period due to the low regrowth rate of wild grass. 2) Proper land for woodland joint cattle grazing is belonging to land of Government ownership and it is very hard to get the permission from office of forestry for cattle grazing purpose. 3) It is also difficult to find a proper time of breeding in grazing area by the difficulty of estrus detection. 4) There are a difficulty to give a proper vaccination and medical examination for the grazing cattle. 3. Improvement plans for woodland joint cattle grazing projects. 1) Obtain sufficient roughages by hoof cultivation and oversowing pasture from the second year of joint cattle grazing period. 2) In order to increase the beef production and to use for a calf production area, Government should arrange that all proper grazing land of Government owned in Kang Won Do convert into woodland joint cattle grazing area. 3) Make a good reproductive record by mixed grazing with a excellent breeding cow in a remote area. And carry out the collective artificial insemination with synchronous puberty induced by injection of puberty stimulation hormone. 4) Make a preventive injection for blackleg, twice medication of fasciola hepatica in a year, and spray and medication of tick insecticide. 4. A policy towards upbringing of woodland joint cattle grazing area. 1) Government should thoroughly investigate about a proper land for woodland joint cattle grazing from all forests. 2) When the area is suitable for the woodland joint cattle grazing, though it is national forest or restricted area, government should make it possible to establish a grazing area. 3) On the proper land foe joint cattle grazing in the remote place, Government should support for the road construction and electric fence equipments by using of national funds. 4) There should be an administrative consideration for well promotion of the project that make woodland joint cattle grazing suitable to the characteristics of Kang Won Do. 5) In order to improve the reproduction record, Government should reform the insufficiency of artificial insemination in the joint cattle grazing area. 6) In order to maintain a proper price of cow, Government should carry out the price plan. 7) When there is any request for grassland formation in the woodland joint cattle grazing area, Government should permit it with preference.