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 %) 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 %) from 12 species detection rates in other species of digenetic trematode metacercariae from various fishes were; Exorchis oviformis, 261 fishes (57 %) from 28 species; Cyathocotyle orientalis, 47 fishes (12 %) from 12 species; Metorchis orientalis, 21 fishes (6 %) from 12 species; Metagonimus yokogawai, 164 fishes (43 %) from 26 species; Pseudexorchis major, 71 fishes (19 %) from 18 species; Metacercaria hasegawai, 77 fishes (20 %) from 25 species; Centrocestus armatus, 24 fishes (6 %) from 7 species; Echinochasmus japonicus, 2 fishes (0.5 %) from 2 species, and unidentified species, 34 fishes (9 %) 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.

A pair of synthetic oligonucleotide primers, designed from the gene encoding a 32-kDa intraerythrocytic piroplasm surface protein of Theileria sergenti, were used to amplify parasite DNA from the blood of T. sergenti-infected cattle by means of the polymerase chain reaction (PCR). PCR-amplified DNA was examined by electrophoresis and by dot blot or microplate hybridization using a parasite-specific cDNA probe. PCR was specific for T. sergenti, since no amplification was detected with DNA from Anaplasma centrale, Babesia ovata, uninfected erythrocytes, and leukocytes. This method was sensitive enough to detect about 4.5 parasites per I.al of blood with a 10-p.l sample volume. Moreover, of 66 specimens from grazing cattle, 40 were microscopically positive, whereas PCR revealed that 54 samples were positive. Therefore, PCR provides a useful diagnostic tool for detecting T. sergenti-infected cattle, and it is significantly more sensitive than the current methods.

ABSTRACT. Benign Theileria species distributed in China and Korea were characterized by allele-specific polymerase chain reaction (PCR), based on the sequences of major immunodominant piroplasm surface protein genes. In China, all the isolates contained Chitose (C) type parasites. One out of 5 isolates tested was a mixed population of Ikeda (I), C and B-2 types, whereas, all the isolates from Korea consisted of I type parasites. Except for 4 isolates, 29 isolates from Korea consisted of more than two types of parasites. The present data showed that benign Theileria species distributed in these countries were mixed parasite populations. -KEY WORDS: allele, PCR, Theileria sergenti/ buffeli/orientalis.

More than 100 cases of canine ehrlichiosis, with three fatalities, were serologically negative by the indirect immunofluorescent antibody (IFA) test with Ehrlichia canis or E. sennetsu antigen but were reactive at titers of 10 to 640 with E. ristcii. Ehrlchia-like agents were isolated from three such cases. The agents isolated from those cases were morphologicalBy indistinguishable from each other and from a prototype, E. risticii, the etiologic agent of equine monocytic ehrlichiosis, in terms of growth characteristics and by light or electron microscopy. The patterns of and products from PCR were identical to those of E. risticii. The 16S rRNA sequences were distinct from those of E. canis and E. ewingu but were identical to those of E. risticii. A PCR product corresponding to the 5' half of the 16S rRNA gene was obtained from amplification of DNA from E. risticii and both sources of the atypical canine ehrlichiosis agent but was not obtained from uninfected host cells. The entire sequence of 719 nucleotides was identical for all three sources. The percentages of relatedness of the partial 16S rRNA gene of the atypical canine ehrlichiosis agent to E. ristcii, E. sennetsu, E. plays, E. equi, E. phagocytophika, E. canis, E.

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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 %) 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 the 3 parts of the river; 53.8 percent in upper, 80.7 % in middle, and 61.0 % 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 %, 29 %, 11 %, 7.9 % and 6.3 %, 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 % and 0.8 %, respectively, out of a total 923 persons.

Anaplasma marginale initial bodies of the Florida strain were purified from infected erythrocytes using a combination of ultrasonic disruption, nonionic detergent and differential centrifugation. Immunochemical analysis revealed at least 12 A. marginale proteins in the molecular mass (m) range 81-15 kDa with a prominent band at 38 kDa. Several of these proteins remained insoluble in the presence of nonionic detergent. Preparations of purified Anaplasma initial bodies contained negligible erythrocytic contamination, as confirmed by the minimal induction of isoantibodies against bovine blood group antigens and the absence of delayed-type hypersensitivity to erythrocytic antigens in immunized animals. A total of 33 crossbred and purebred Holstein cattle were vaccinated with either 1.5, 1.0, or 0.1 mg protein of intact initial bodies, or with 1.0 mg of solubilized Anaplasma protein. The immunogens were supplemented with 3.0 mg Quil-A saponin adjuvant and administered in 2 subcutaneous injections given at a 4-week interval. A similar number of nonvaccinated cattle served as controls. Three months after vaccination, all cattle were challenged by inoculation of 10(9) virulent A. marginale of either the homologous (Florida) or heterologous (Venezuelan) strains. Vaccinated cattle showed solid protection after homologous and heterologous challenge, characterized by parasite clearance and minimal hematocrit reductions. Initial data from four field vaccine trials revealed a reduced incidence of clinical anaplasmosis among immunized animals. Use of immunogens consisting of purified A. marginale initial bodies offers a potential immunoprophylactic approach to control of bovine anaplasmosis.