Chronologic change of serum IgG antibody response in chickens reinfected with Cryptosporidium baileyi

Eight 2-day-old SPF chickens were each inoculated orally with a single dose of 5+O105 oocysts of Cryptosporidium baileyi, and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. baileyi IgG antibody levels remained high(1:106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1+O107 oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation(PCI), at 1:1,024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles, but did not exhibit any oocyst shedding before or after experimental reinfection.

The wormicidal substances of fresh water fishes on Clonorchis sinensis VII. The effect of linoleic acid and ethyl linoleate on parasite viability

In an attempt to analyze the clonorchicidal activity of linoleic acid and ethyl linoleate in vitro, the wormicidal effects on Clonorchis sinensis were chronologically monitored in dose titration experiments. Excysted metacercariae were killed within a period of 31.0+-4.0 min, 149.3+-4.l min and 207.0+-13.5 min with 100.0 mg, 0.1 mg and 0.001 mg linoleic acid, respectively. The time required for the linoleic acid to kill adult worms was 167.0+-0.8 min with 100.0 mg, 253.0+-0.8 min with 0.1 mg, and 277.0+-0.8 min at 0.001 mg titration. Clonorchicidal activity of ethyl linoleate was relatively delayed as death was observed within 263.3+-2.9 min, 286.0+-0.5 min, and 318.0+-0.8 min for 100.0 mg/ml, 0.1 mg/ml and 0.001 mg/ml concentrations, respectively. The wormicidal effects observed with these pure anticlonorchal substances were found to be similar to the biological activity of native products derived from the mucus of the fresh water fish.

The karyotype of Fischoederius cobboldi (Poirier, 1883) from Korean cattle

As a series of systematic classification of paramphistomes, the worms in the rumen and reticulum of 310 Korean cattle slaughtered at Chonju abattoir were collected from February 1986 to June 1987 and were classified by morphology of the worms. Afterwards, the karyotype of Fischoederius cobboldi (Poirier, 1883), which is a very rare species in Korean cattle, was studied with germ cells of the worm by means of modified air-drying method. The chromosome numbers in the haploid and diploid cells of 315 F. cobboldi were n=9 and 2n=18, respectively. The meiotic divisions were observed frequently; 1,904 haploid and 49 diploid cells were recognized. Nine pairs of mitotic chromosomes were homologous in the metaphase stage and the chromosomes were composed of seven medium-sized metacentrics (m) or submetacentrics (sm) and two small-sized submetacentrics (sm). While, meiotic metaphases were composed of seven medium and two small-sized chromosomes. The 3rd, 4th, 2nd and 5th pairs of chromosomes was metacentric having centromere indices of 40.4 per cent, 40.0 per cent, 39.7 per cent and 38.9 per cent, respectively, and the remaining ones were submetacentric with centromere indices from 32.4 per cent to 36.2 per cent. As a series of C-banding method, C-band was shown in centromeric region from all of the haploid germ cells, except chromosome No. l which included heterochromatin at the tip region. Chromosomes No. 4, 6 and 8 showed remarkable C-band distinguished from others.

The karyotype of Paramphistomum cervi (Zeder, 1790) from Korean cattle

As a series of systematic classification of paramphistomes, the worms in the rumen and reticulum were collected on 214 Korean cattle slaughtered at Jeonju abattoir from January 1986 to April 1987 and were classified by means of morphology. Afterwards, the karyotype of Paramphistomum cervi (Zeder, 1790) was detected by means of modified air-drying method from germ cells of the worms. The results were summarized as follows: In the chromosome number of 254 P. cervi, the haploid cell was n=9 and the diploid 2n=18. The meiotic divisions were observed frequently; 1,924 haploid and 32 diploid cells were reliable. Nine pairs of mitotic chromosomes were homologous in the metaphase stage, and the chromosomes were composed of five medium-sized metacentrics (m), subtelocentrics (st) or submetacentrics (sm) and four small-sized subtelocentrics(st) or submetacentrics(sm). Meiotic metaphase was composed of five medium and four small chromosomes in size. As a series of C-banding method, C-band was showed in centromeric region from all of the haploid germ cells. Whereas chromosome No. 3 and 5 included heterochromatin on the tip region, chromosome No. 4 on the distal region and No. 6 proximal region. And chromosomes No. 2 and 8 showed a remarkable C-band distinguished from other chromosomes.

Karyotype of Fasciola sp. obtained from Korean cattle

As a series of systematic classification for Korean common liver fluke, Fasciola sp., karyotype was investigated by means of the modified air-drying technique and of the regular Giemsa staining, Also, C-staining method was applied for detailed karyological analysis from the germ cells of the fluke. The following is a brief summary of the leading facts gained through the experiment. Korean Fasciola sp. was classified into three types based on their chromosomal complements; individuals with 20 or 30 chromosomes and with a 20/30 mosaic constitution. Worms having 30 chromosomes represent a triploid form with 3 sets of 10 basic chromosomes, while those with 20 chromosomes were diploid and mosaic individuals were 2n/3n mixoploid. The frequency of the individual type calculated is as follows; 67.45% of 212 flukes examined was of diploid, 10.85%, triploid, and the rest, 21.7%, mixoploid, respectively. In many cases, two or three types were found in the peculiar bovine host while single type inhabitant was about 20% out of 52 cases. The twenty chromosomes consisted of 1 parir of large metacentrics, 4 pairs of medium-sized subtelocentrics, and 5 pairs of small submetacentrics, while constitution of the thirty chromosomes was nearly interpreted as a triploid form with 3 sets of 10 basic chromosomes. The high centromeric indexes of both types are the first pairs among all the examined, and 37.93% was of diploid and 47.93%, triploid, respectively. In mixoploid individuals, constitution of the chromosomes of diploid or triploid cells was the same as that of diploid or triploid individuals. All the chromosomes of the germ cells in both types showed C-band around the centromeic region and especially the chromosomes no's 3, 7, and 8 showed a remarkable C-band distinguished from other chromosomes. The variance for the sizes of the worms and the eggs were not parallel with three different genotypes in Korean common liver fluke.

The karyotype of Paramphistomum explanatum (Creplin, 1849) obtained from Korean cattle

As a series of systematic classification of paramphistomes, in the first step, paramphistomes in the rumen and reticulum were collected on 170 Korean cattles (2-3 years age, male) slaughtered at Jeonju abattoir from July 1984 to September 1985 and were classified by means of morphology of the worms. Afterwards, the karyotype of Paramphistomum explanatum (Creplin, 1849) which is the common in Korean cattle was detected by means of modified air-drying method from testis cells of the worm. The following is a brief summary of the leading facts gained through the experiment. Most of the cattle slaughtered at the abattoir were infected with paramphistomes. The 5 species of the worms were detected on 170 Korean cattle and the worm burden per head was from 2 to 784 (on the average 170) worms, 120(70.59 percent) heads out of them involving 2-100 worms. In 28,900 individuals of paramphistomes obtained on 170 Korean cattle, appearance rates of various worms were as follows : 49.74 percent in P. explanatum, 48.08 percent in P. cervi, 0.98 percent in Orthocoelium orthocoelium, 0.89 percent in Fischoederius cobboldi and 0.14 percent in Cotylophoron cotylophorum. The chromosome number of 620 P. explanatum in the haploid and diploid cells was n=9 and 2n=18, and abundant cells in meiotic division were observed; 1,420 haploid and 38 diploid cells were reliable. Nine pairs of mitotic chromosomes were homologous and the chromosomes were composed of five medium-sized metacentrics (m), subtelocentrics (st) or submetacentrics (sm)and four small-sized subtelocentrics (st) or submetacentrics (sm), while meiotic metaphase chromosomes were composed of five medium and four small-sized. The haploid of the testis cells showed C-band in the centromeric region from 8 of them, whereas the remaining chromosome No. 5 included heterochromatin on the tip region, and chromosomes No. 3 and No. 7 showed a remarkable C-band distinguished from other chromosomes.

Fractionation of antigen for ELISA of bovine fascioliasis

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: The specificity (95 per cent confidence interval in negative sera of bovine fascioliasis ; Mean+2 x SD of absorbance ) of the first (MW>150,000) and the second antigens (MW 120,000) were 93.7 per cent, but those of others including crude antigen showed 100 per cen.t. 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 per cent, 87.5 per cent and 0 per cent, respectively, but those of others showed all 100 per cent. 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. 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 forth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. The wheal size for the bovine infected with F. hepatica was 2.46+-0.15 cm 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 forth, 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.

Trachelobdella sinensis Blanchard, 1896 found from Cyprinus carpio nudus in Korea

Trachelobdella sinensis was found in Cyprinus carpio nudus. The morphological chararacteristics were described. It is the first record of Trachelobdella sinensis in Korea(summarized from text).

A record of Diplozoon nipponicum Goto, 1891 found from Cyprinus carpio nudus in Korea

Diplozoon nipponicum, a monogenean trematode from Cyprinus carpio was described according to the characteristic findings of X-form adult, metrical data, and presence of a pair of sticky glands. It is the first record of the presence of Diplozoon nipponicum in Korea.

Experimental studies on the second intermediate hosts of Clonorchis sinensis IV. Observations on the fate of metacercariae of Clonorchis sinensis in the fish host, Cultriculus eigenmanni

In order to judge appropriately the suitability as the second intermediate host of Clonorchis sinensis, the present experiments were proposed to survey the metacercaria of C. sinensis in the fish host, Cultriculus eigenmanni, in the field, and to observe, by experimental infection of the fish with Clonorchis cercaria, the penetrating ability of the cercaria, maturity, the process of degeneration and extinction, and infectivity of the metacercaria. The following is a brief summary of the leading facts gained through the experiments. All the ten fish (mean body weight 13.4 +/- 3.43 g) which were caught in Ok-ku irrigation reservoir were infected with the metacercariae and the average number of the metacercariae found in the fish body/gram was 4.55. The fish was easily subject to invasion of the cercaria, and the examination of the cercariae after having the fish infected disclosed that 24 hours after the infection, all of the cercariae formed their cysts in muscle and the metacercariae kept growing, that 7 days later the metacercariae were found folding their bodies twice, and that in 15 days the characteristic structure of the metacercariae was complete and made a vigorous rotary movement intermittently. In the case of control fish, Pseudorasbora parva, both penetrating ability of the cercariae and the infective degree of the metacercariae to control host were stronger as compared with the fish host because that the control fish had higher suceptibility to C. sinensis. Afterwards, the metacercariae in the fish host came to a state of maturity and beyond this stage some metacercariae started the process of degeneration and extinction in as early as 30 days, and as more days elapsed, mortality of the metacercariae was not uniformly increased but difference of mortality appeared to be according to the individual of fish host. That is, in 390, 540 and 586 days, all of the metacercariae in the fish hosts became dead, meanwhile almost all of those found in the fish host of 596 days after the infection survived (mortality; 8.51 percent). Percentage of orally administered metacercariae developing to maturity in rabbit is 10.46 percent in the last case. As the days were progressed the shapes of the metacercariae in the fish host tended to change from ellipsoid to almost round shapes, the length became shorter while the width was wider. The results shown above revealed that C. eigenmanni was, though it could not be proper host than P. parva, suitable as the second intermediate host in addition to our previous observation results.

Application of micro-ELISA in serodiagnosis of fascioliasis in cattle

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 Fasciola hepatica, peroxidase of conjugate anti-cattle IgG and orthophenylenediamine as a substrate by micro-method technique of Voller et al. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows: In assay for optimal dilution of stock antigen, the antigen (protein contents; 0.8 mg/ml) was diluted from 1/50 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. In assay for optimal dilution of sera, the sera were diluted from 1/25 to 1/400 with in PBS/ 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). In the 27 cases of negative intradermal test, OD values of the ELISA are 0.447 +/- 0.144, the 95 percent confidence interval (Mean+2 x 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 percent 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. 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. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was 0.846+0.224(-0.224). The 75 percent (44 cattle) among them had higher value with compared to the criterion, and the 60 percent (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. Prevalence of fascioliasis was 43.4 percent in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

The wormicidal substance of fresh water fishes on Clonorchis sinensis V. Purification and chemical characterization of clonorchicidal substance from epidermal mucus of Cyprinus carpio

As a series of studies to clarify clonorchicidal substances in body surface mucus of some fresh-water fishes, the substance in the epidermal mucus of Cyprinus carpio was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature. Wormicidal trial was done in vitro, and the results obtained are summarized as follows: The mucus was extracted by ethyl ether and separated into 4 fractions by column chromatography using benzene as solvent. The second fraction with yellowish red colour among them showed the strongest clonorchicidal effect. The yellowish red fraction obtained by column chromatography was then fractionated into 6 spots by thin layer chromatography with petrol. ether/chloroform(30/70, v/v), and the Rf. 0.714 spot among the 6 spots showed the strongest effect. The Rf. 0.714 spot was further fractionated into 6 spots by thin layer chromatography with benzene/acetone (90/10, v/v), and the Rf. 0.800 spot among the later 6 spots revealed the strongest effect. The Rf. 0.800 spot was chromatographed on column with benzene and 2 fractions were obtained. The second fraction of light brown colour represented the final purified fraction. By these purification procedures, clonorchicidal substance was purified 15-fold with 0.03 percent yield from the mucus of C. carpio, and 10mg of the final fraction killed the cercaria in 26 min, the metacercaria in 115 min, and the adult in 443 min. Infra red and nuclear magnetic resonance spectrometric analysis of the purified substance revealed that the substance belongs to an ethyl ester of unsaturated fatty acid with 2 double bonds, 15 methylene groups and l methyl group.

Survey on encysted cercaria of trematodes from fresh-water fishes in Tongjin riverside areas in Korea

In an attempt to clarify the epidemiological feature of distomiasis in Tongjin riverside area, the prevalence of distomiasis in the residents and infection rates of the metacercariae in fresh-water fishes were investigated at the upper, middle and lower reaches of the river from January to April 1984. The results obtained were summarized as follows: Out of a total of 931 fresh-water fishes which composed of 33 different species, 611 fishes (65.6 percent) of 31 species were found positive with digenetic trematode metacercariae of 16 different species, and there were some differences in infection rates of the metacercariae among the fishes in the3 parts of the river; 53.8 percent in upper, 80.7 percent in middle, and 61.0 percent in lower reaches, respectively. Infection rates of the metacercariae of Exorchis oviformis, Metagonimus yokogawai, Echinochasmus japonicus, Metorchis orientalis and Clonorchis sinensis in the fishes were 48 percent, 29 percent, 11 percent, 7.9 percent and 6.3 percent, respectively. The average number of the encysted larvae of Clonorchis found in fish body/gram showed 4.44 in Pseudorasbora parva, Gnathopogon coreanus (1.2), Microphysogobio yaluensis(0.76), Abbottina springeri(0.4), Acanthorhodeus asmussi (0.21) and Cultriculus eigenmanni (0.17), respectively. The average number of the metacercariae of Metagonimus found in fish body/gram disclosed 34.01 in Zacco platypus, Zacco temmincki (16.46), Carassius carassius (5.35), Moroco oxycephalus (1.54), Aphyocypris chinensis (1.5) and etc., respectively. Detection rates of the eggs of Clonorchis and Metagonimus among residents were 1.1 percent and 0.8 percent, respectively, out of a total 923 persons.

Experimental Infection of Clonorchis sinensis to Cyprinus carpio nudus

As a part of observation on the suitability as a second intermediate host of Clonorchis sinensis, various infection experiments were tried to Cyprinus carpio nudus. The results obtained were summarized as follows: The cercariae which attempted to contact with Cyprinus carpio nudus in the water were observed under the stereomicroscope. After contact, cercariae began to separate its tails from the bodies at 4 minutes increasingly, and then the number ranged to 80 percent at 13 minutes after the encounter. But very few cercariae could actually invade into the epidermis of the fish. When Cyprinus carpio nudus were exposed to a number of cercariae in the beaker, only a few cercariae could invade through the epidermis. Most of the invaded cercariae were killed before forming the cyst. Also rare encysted cercariae were found to be dead within 48 hours. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in the indoor aquarium. When observed after 12 days, to determine the susceptibility, no metacercaria was found infected. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in Kimhae lake for 2 months in summer of 1982, in order to check the natural infection. No metacercaria of Clonorchis sinensis was found from the fish. In the cases of control fish, Pseudorasbora parva, numerous cercariae were inspected on the course of invasion through epidermis of the fish under the stereomicroscope. And many metacercariae were also found from the fish whenever they were kept in beaker, indoor aquarium, or the lake for natural infection. The results suggested that Cyprinus carpio nudus might not be proper intermediate host of Clonorchis sinensis.

The wormicidal substance of fresh water fishes on Clonorchis sinensis IV. Preliminary research on the wormicidal substance from mucus of Cyprinus carpio nudus

As a series of studies on the suitability as a second intermediate host of Clonorchis sinensis, the wormicidal effects of mucus from epidermis of Cyprinus carpio nudus were observed. The results obtained were summarized as follows: On the silica gel thin layer chromatography with the greenish yellow supernatant in acetone:benzene/10:90 as carrier, seven spots were observed in iodine chamber. The spot of Rf. 0.225 value among them disclosed the strongest wormicidal effect on the cercaria, the excysted metacercaria, and the adult of C. sinensis. Meanwhile, no seasonal variations were found in Rf. values and effect of the wormicidal fractions. The wormicidal materials in epidermal mucus of C. carpio nudus and Carassius carassius were compared in their effect, and the former was more effective killing substance than the latter. It seemed that C. carpio nudus could not be proper intermediate host of C. sinensis according to results of the present study, in addition to our previous observation results.

Epidemiological studies of Clonorchis sinensis in Mangyeong riverside areas in Korea

In an attempt to clarify the epidemiological feature of C. sinensis in Mangyeong riverside area, the prevalence of clonorchiasis, infestation rate of the cercariae in Parafossarulus manchouricus, and detection rate of the metacercariae in fresh-water fishes were investigated from March 1 to September 30, 1983 at the upper, middle and lower reaches of the river. The results obtained were summarized as follows:Detection rate of C. sinensis egg among inhabitants was 8.2 percent out of a total of 1,266 persons. but the differences in detection rates were not statistically signifcant among upper, middle and 1ower reaches. According to sex, the detection rates were 10.3 percent in male and 6.1 percent in female (p<0.05), but by age groups, increases of the rates were observed as increase in age (p<0.05). Out of a total of 380 fresh-water fishes of 32 different species, 93 fishes (25 percent) of 12 species were found positive with Clonorchis metacercariae, and there were differences in infection rates of the metacercariae among the fishes in 3 parts of the river; 11 percent in upper, 35 percent in middle, and 34 percent in lower reaches respectively. The metacercarial detection rates from various fishes were 97 percent in Pseudorasbora parva, Cultriculus eigenmanni (85 percent), Gnathopogon strigatus (67 percent), Microphysogobio yaluensis (50 percent), Gnathopogon coreanus (47 percent), Pungtungia herzi(44 percent), Abbottina rivularis (40 percent), Moroco oxycephalus (33 percent), Coreoleuciscus splendidus (32 percent), Gnathopogon majimae (26 percent), Rhodeus ocellatus (7 percent), and Aphyocypris chinensis (3 percent) respectively. Although very few P. manchouricus were collected at upper reach, 12 snails (0.7 percent) among a total of 1,713 were found infected with Clonorchis cercariae. Also the cercariae of Echinochasmus japonicus (7.99 percent), Lexogenes liberum (0.99 percent), Cyathocotyle orientalis (0.75 percent), Exorchis oviformis (0.23 percent) and Asymphylodora japonica (0.05 percent) were detected from the snails.

Survey on encysted cercariae of trematodes from fresh-water fishes in Mangyeong riverside area

In order to clarify infestation pattern for the encysted larvae of digenetic trematodes from fresh-water fishes, this survey was carried out from March to September, 1983. A total of 380 fishes of 32 species were collected with netting at the three reaches, upper, middle and lower in Mangyeong riverside area. After the fishes were dissected into small scraps, they were pressed under cover glass and examined for the presence of those of digenetic trematodes with a microscope. The results obtained were as follows: Out of a total of 380 fishes inspected, 320 fishes (84 percent) from 31 species were found positive with digenetic trematode metacercariae; more than 10 species of the metacercariae were detected in Pseudorasbora parva; Gnathopogon majimae, Microphysogobio yaluensis, Cultriculus eigenmanni and Gnathopogon coreanus (more than 8 species); Aphyocypris chinensis(8 species) and etc. respectively. Clonorchis sinensis metacercariae were found positive from 93 fishes (25 percent) from 12 species detection rates in other species of digenetic trematode metacercariae from various fishes were; Exorchis oviformis, 261 fishes (57 percent) from 28 species; Cyathocotyle orientalis, 47 fishes (12 percent) from 12 species; Metorchis orientalis, 21 fishes (6 percent) from 12 species; Metagonimus yokogawai, 164 fishes (43 percent) from 26 species; Pseudexorchis major, 71 fishes (19 percent) from 18 species; Metacercaria hasegawai, 77 fishes (20 percent) from 25 species; Centrocestus armatus, 24 fishes (6 percent) from 7 species; Echinochasmus japonicus, 2 fishes (0.5 percent) from 2 species, and unidentified species, 34 fishes (9 percent) from 15 species respectively. The sums of average number of the encysted larvae of all species found in fish body/gram showed 83 in P. parva, Cobitis taenia (74.2), A. chinensis (28.5), Pseudoperilampus uyekii (26.6), G. majimae (19.6) and etc. respectively and the average peak number of each metacercaria in fish body/gram showed 21.7 C. sinensis, 24 E. oviformis , 15.3 M. orientalis and 6.1 E. japonicus in P. parva ; 42.7 C. orientalis and 25.1 M. yokogawai in C. taenia; 8.3 C. armatus and 8.3 M. hasegawai in P. uyekii; 6.3 P. major in Carassius carassius, and 2.9 unidentified species in G. majimae respectively.

Clavate Cells Of Epidermis In Cyprinus Carpio Nudus With Reference To Its Defence Activity To Clonorchis Sinensis

No abstract available.

Localization and isolation of anthelmintic activity on Clonorchis sinensis from stem cortex of Machilis thunbergii

The present work was to localize and isolate the anthelmintic substance from stem cortex of Machilis thunbergii which grows in all parts of the Korea peninsula. The dried M. thunbergii bark was extracted with a solvent mixture (Methanol: Ethyl acetate/1:1) by a Soxhlet extractor at 70C for 24 hours, and then the extract was separated into solid matter (negative wormicidal) and benzene extract (positive wormicidal) by benzene solvent. The anthelmintic action was observed on the cercaria, the liberated metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with a spot of benzene extract in benzene solvent, nine spots were observed in iodine chamber and the spot of Rf. 0.16 value of them appeared the most strong wormicidal effect on the cercaria and the liberated metacercaria of C. sinensis. Afterward, a 0.3 g colourless solid substance which was equivalent to the Rf. 0.16 value spot was isolated from the benzene extract of 3 kg M. thunbergii stem cortex by means of silica gel column chromatography using benzene carrier, and the adult of C. sinensis was killed within 6 minutes by 0.05 percent solution of the substance.

Isolation of anthelmintic substance on Clonorchis sinensis from tuber of Bletilla striata

The present work was done to isolate the anthelmintic substance from the tuber of Bletilla striata which grows Gangweon-do of the Korea peninsula. The dried B. striata tuber was extracted with ethyl ether by a Soxhlet extractor at 40C for 24 hours, and then the extract was separated into insoluble material (negative wormicidal) and soluble material (positive wormicidal) by petroleum ether solvent. The wormicidal substance was separated in the silica gel column chromatography from the petroleum ether extract, using benzene (90 percent) and acetone (10 percent) as a solvent. The anthelmintic activity was observed in vitro on the cercaria, the excysted metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with the wormicidal substance in benzene:acetone/90:10, five spots were observed in iodine chamber and the spot of Rf. 0.18 value of them appeared the most strong wormicidal effect on the cercaria of C. sinensis. Afterward, a substance which was equivalent to the Rf. 0.18 value spot was isolated from the wormicidal substance by means of silica gel column chromatography using 1 part of ethyl acetate and 6 parts of chloroform carrier, and the excysted matacercaria and the adult of C. sinensis were killed within 14 minutes and 128 minutes in the substance.