Metagonimus miyatai n. sp. (Digenea: Heterophyidae) is described based on adult flukes collected from experimental dogs and hamsters fed with metacercariae encysted in the sweetfish, Plecoglossus altivelis, dace, tribolodon hakonensis and T. taczanowskii, common fat-minnow, Morocco steindachneri, pale chub, Zacco platypus, and dark chub, Zacco temmincki, and on those collected from naturally infected humans. The new species was morphologically compared with M. yokogawai and M. takahashii obtained from experimental animals fed with the sweetfish and the crucian carp, Carassius carassius, respectively. The uterine loops of M. miyatai reached near the posterior end of the body through the space between the two testes, whereas those of M. yokogawai, occupied only the space between the acetabulum and anterior border of two testes. This uterine tubule distribution was similar to that of another closely related species, M. takahashii. However, vitellaria of M. miyatai ended in front of the posterior end of the left testis, while those of M. takahashii reached the posterior end of the left testis and ran it over. By raising M. miyatai as a new species, differentiation of M. yokogawai and M.
takahashii became very clear. A key to the species of the genus Metagonimus in the Far East has been proposed.
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Genus Metagonimus has been a subject of taxonomic debates for several years. In morphological aspects, M. yokogawai has been thought to have three subtypes, namely Yokogawa type (M. yokogawai in strict sense), Miyata type and Koga type, But differences in the intestinal pathology induced by these subtypes have not been studied yet. In this study we compared the pathological reactions induced by M. yokogawai and Metagonimus Miyata type using proliferating cell nuclear antigen (PCNA) index. Metacercariae (Mc) of M. yokogawai were collected by artificial digestion of Plecoglossus altivelis and Mc of Metagonimus Miyata type were collected from Zacco platypus. Three hundreds Mc of each species were infected orally to ICR mice. The mice were sacrificed at 3, 6, 10, 16 and 23 days after infection and the small intestines were resected into three portions (proximal, middle, and distal). Immunohistochemical staining for PCNA was done using PC-10 (DAKO-PCNA, CA, USA). The PCNA indices in M. yokogawai infected group on the 6th and 23rd day after infection were lower than in the control and Miyata type infected groups (p < 0.05) from all of the three intestinal regions. On the other hand, the control group and Metagonimus Miyata type infected group did not make any differences in PCNA indices. The villus/crypt (V/C) ratio was also decreased significantly in M. yokogawai infected mice but not in Metagonimus Miyata type infected ones. It is suggested that M. yokogawai induce villous atrophy through a decrease in the cell proliferation at the crypt. The results of this study suggested that M. yokogawai induce more serious intestinal pathology than Metagonimus Miyata type.
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We tried to compare the three kinds of Metagonimus species, M. yokogawai, Metagonimus Miyata type, and M. takahashii, which were known to be distributed in Korea with polymerase chain reaction based-restriction fragment length polymorphism (PCR-RFLP) patterns. We amplified the internal transcribed spacer 1 (ITS1) site of ribosomal RNA and mitochondrial cytochrome c oxidase I (mCOI) gene. The restriction patterns of ITS1 gene with Rsa I, Alu I and Msp I showed multiple fragmented bands of different sizes between three kinds of Metagonimus. In case of mCOI gene, Rsa I and Alu I enzymes produced differentially fragmented band patterns. According to the parsimony analysis of PCR-RFLP patterns, the estimated genetic divergence between M. yokogawai and Metagonimus Miyata type was 0.034880, between Metagonimus Miyata type and M. takahashii was 0.028098, between M. yokogawai and M. takahashii was 0.018179. It is suggested that Metagonimus Miyata type may be separate species and evolutionize at the older time than the other two species.
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