Morphological and molecular characterization of Austrodiplostomum compactum metacercariae in the eyes and brains of fishes from the Ivaí River, Brazil / Caracterização morfológica e molecular de metacercárias de Austrodiplostomum compactum em olhos e cérebros de peixes do Rio Ivaí, Brasil

Abstract Austrodiplostomum spp. (Platyhelminthes: Digenea) are endoparasites with a broad geographic distribution in South America. During the larval stage, they parasitize the eyes, brains, muscles, gill, kidneys and swim bladder of a wide variety of fishes. The metacercariae of Austrodiplostomum spp. have several morphological characteristics during development, but are very similar among species, which makes it necessary to use molecular tools to contribute to the elucidation during the larval stage. The objective of this study was to perform morphological and molecular analyses of Austrodiplostomum sp. found in specimens of Hypostomus sourced from the Ivaí River in the state of Paraná, Brazil. Of the 93 analyzed specimens (H. hermanni [n = 50], H. albopunctatus [n = 9], Hypostomus sp. 1 [n = 24], and Hypostomus sp. 2 [n = 10]), 60 were parasitized. A total of 577 Austrodiplostomum sp. metacercariae was collected from the infected hosts; DNA from seven of these samples was extracted, amplified, and sequenced. The morphological data associated with the genetic distance values and the relationships observed in the COI gene tree, indicate that all metacercariae were A. compactum. This is the first record of A. compactum parasitizing H. hermanni, H. albopunctatus, Hypostomus sp. 1, and Hypostomus sp. 2 in the Ivaí River.

Resumo Austrodiplostomum spp. (Platyhelminthes: Digenea) são endoparasitos com uma ampla distribuição geográfica na América do Sul. Durante a fase larval, parasitam os olhos, cérebros, músculos, brânquias, rins e bexiga natatória de uma grande variedade de peixes. As metacercárias de Austrodiplostomum spp. apresentam várias características morfológicas durante o desenvolvimento, as quais são muito semelhantes entre as espécies, o que torna necessário o uso de ferramentas moleculares para contribuir para a elucidação durante a fase larval. O objetivo deste estudo foi realizar análises morfológicas e moleculares de Austrodiplostomum sp. encontradas em espécimes de Hypostomus provenientes do rio Ivaí, no Paraná, Brasil. Dos 93 espécimes analisados (H. hermanni [n = 50], H. albopunctatus [n = 9], Hypostomus sp. 1 [n = 24], e Hypostomus sp. 2 [n = 10]), 60 foram parasitados. Um total de 577 metacercárias de Austrodiplostomum foram coletadas dos hospedeiros infectados; o DNA de sete dessas amostras foi extraído, amplificado e sequenciado. Os dados morfológicos, associados aos valores de distância genética e as relações observadas na árvore gênica do COI, indicam que todas as metacercárias são A. compactum. Este é o primeiro registo de A. compactum parasitando H. hermanni, H. albopunctatus, Hypostomus sp. 1, e Hypostomus sp. 2 no rio Ivaí.

Distribution of cysts holding Ascocotyle (Phagicola) longa metacercariae in tissues and organs of mugilid / Distribuição de cistos contendo metacercárias de Ascocotyle (Phagicola) longa em tecidos e órgãos de mugilídeos

Abstract Ascocotyle (Phagicola) longa is an etiological agent of human phagicolosis. Mugilids are the second intermediate host, the first being Heleobia australis, and mugilids predatory birds and mammals are its definitive hosts. The occurrence of cysts holding A. longa metacercariae is described in mugilids with a prevalence of up to 100%. The wide geographical distribution of A. longa and its intermediate hosts coupled with the rise in the consumption of raw or poorly cooked fish may elevate the risk of human infection. Therefore, in this study, we aimed to verify the distribution pattern of cysts holding A. longa in mugilids. The tissue and organ samples of these fish were processed in a domestic blender and examined under a stereoscopic microscope to identify the cysts holding the digenetic metacercariae. Of the 24 (100%) fish samples that were analyzed, 12 of Mugil curema and 12 of Mugil liza possessed cysts holding A. longa metacercariae. Digenetic cysts were identified to be present in the gills, heart, stomach, liver, intestines, mesentery, and muscular tissues collected from M. curema and M. liza. Conclusively, in M. curema, the cysts holding A. longa metacercariae were found to be distributed randomly throughout the fish body in almost every tissue and organ that was examined.

Resumo Ascocotyle (Phagicola) longa é o agente etiológico da fagicolose humana. Os mugilídeos são os segundos hospedeiros intermediários. O primeiro é Heleobia australis e pássaros e mamíferos predadores de mugilídeos, os hospedeiros definitivos. A ocorrência de cistos contendo metacercárias de A. longa é descrita em mugilídeos, com até 100% de prevalência. A ampla distribuição geográfica de A. longa e seus hospedeiros intermediários, acompanhado do aumento do consumo de peixe cru ou mal cozido, pode aumentar o risco de infecção humana, portanto o objetivo do presente estudo foi verificar o padrão de distribuição de cistos, contendo metacercárias de A. longa, em mugilídeos. Amostras de tecidos e órgãos desses peixes foram processadas em liquidificador doméstico e observadas em microscópio estereoscópico à procura de cistos contendo metacercárias do digenético. Todos os 24 (100%) mugilídeos, 12 Mugil curema e 12 Mugil liza examinados apresentaram cistos contendo metacercária de A. longa. Foram observados cistos do digenético nas brânquias, coração, estômago, fígado, intestino, mesentério e tecido muscular de M. curema e de M. liza. Em M. curema, os cistos contendo metacercárias de A. longa estão distribuídos de forma aleatória por praticamente todos os tecidos e órgãos dos mugilídeos examinados.

First report of Diplostomidae metacercariae (Trematoda: Digenea) in African catfish Clarias gariepinus (Siluriformes: Clariidae) in Brazil / Primeiro relato de metacercárias de Diplostomidae (Trematoda: Digenea) em bagre africano Clarias gariepinus (Siluriformes, Clariidae) no Brasil

Abstract Clarias gariepinus is a fish from North of South Africa and was later introduced in several countries, including Brazil. The present study aimed to describe the first report of Diplostomidae metacercariae in C. gariepinus in Brazil. For this, 30 C. gariepinus were captured in a lake in the city of Campos dos Goytacazes, RJ, Brazil. Fishes were euthanized using freezing and necropsied for collection of parasites. The organs were dissected and analyzed for the presence of parasites that were processed for light and scanning electron microscopy. Trematodes were stained with Semichon's Carmine and Gomori's Trichrome, observed and schematized under a light microscope with image analysis software. A total of 190 trematodes were collected from the gills, suprabranchial organs, heart, stomach, intestinal mesentery, liver and body cavity of the fish. The parasites had a foliaceous body divided by a discrete constriction, without genital primordia, and a holdfast organ present at the posterior region, typical of metacercariae of the family Diplostomidae. It was classified as the 'Diplostomulum' morphotype, based on the morphology of the reserve bladder structure. This is the first report of the metacercariae of Diplostomidae parasitizing C. gariepinus in the Americas. This fish acts as an intermediate or paratenic host of this digenean in Brazil.

Resumo Clarias gariepinus é um peixe do Norte da África do Sul e posteriormente introduzido em diversos países, incluindo o Brasil. O presente estudo teve como objetivo descrever o primeiro relato de metacercárias em C. gariepinus no Brasil. Para tanto, 30 C. gariepinus foram capturados em um lago na cidade de Campos dos Goytacazes, RJ, Brasil. Os peixes foram eutanasiados por congelamento e necropsiados para a coleta de parasitos. Os órgãos foram dissecados e analisados para a presença de parasitos que foram processados para microscopia ótica e electronica de varredura. Os Trematodeos foram corados com carmim de Semichon e tricrômico de Gomori, observados e esquematizados sob microscópio ótico equipado com software de análise de imagens. Um total de 190 trematodeos foram coletados das brânquias, órgãos suprabrânquiais, coração, estômago, mesentério intestinal, fígado e cavidade corporal dos peixes. Os parasitos tinham corpo foliáceo, dividido por uma discrete contrição, sem primórdio genital, e órgão tribocítico presente na região posterior, típico das metacercárias da fámilia Diplostomidae. Foram classificados como do morfotipo 'Diplostomulum', baseado na morfologia da estrutura da bexiga de reserva. Este é o primeiro relato de metacercárias de Diplostomidae parasitando C. gariepinus nas Américas. Este peixe atua como hospedeiro intermediário ou paratênico deste Digenea no Brasil.

Parasitic fauna of Hyphessobrycon takasei and Hyphessobrycon amapaensis (Osteichthyes: Characidae), ornamental fish of two basins from Amapá state, Brazil

This study compared the parasites fauna of Hyphessobrycontakasei and H. amapaensisfrom hydrographic basins in Amapá state, Brazil. Of the 42 H. takasei examined, 95.2% were parasitized by one or more species such as Dactylogyridae gen. sp., encysted metacercariae of digenean, adults and larvae of Procamallanus (Spirocamallanus) inopinatus and larvae of Camallanus sp. A total of 224 parasites were collected from H.takasei, and there was a dominance of nematode species and dispersion pattern aggregated of parasites. The species richness of parasites varied of 0 to 3 species, Brillouin index of 0-0.97 and evenness of 0-0.70. Of the 32 specimens of H. amapaensis examined, 28.1% were parasitized by Camallanus sp. larvae, which presented random dispersion pattern. For H. takasei, no correlation of host length with Brillouin diversity and species richness of parasites was found. The presence of these nematode species and metacercariae of digenean indicates that these fish may act as intermediate hosts. This is the first study to gather information about the parasite fauna of these endemic fish from the eastern Amazon, providing records that contribute new reports on occurrence of parasite species in new localities. This study on parasites of H. takasei and H. amapaensis may help prevent the introduction of parasites in other regions of the country, as well as in other countries due to aquarium fish.Since the ornamental fish exportation from the Brazil is largely dominated by wild Amazonian species, biosecurity export conditions should be require all exported fish to be inspected and certified to show no clinical signs of parasite and diseases. Lastly, this knowledge on the parasitic epidemiology may help improve the health and quality of these fish being exported from the Amazon region, reducing losses on all stages of this production chain.

Metacercariae of Austrodiplostomum spp. (Digenea: Diplostomidae) infecting the eyes and brains of fish in Brazilian Amazon / Metacercaria de Austrodiplostomum spp. (Digenea: Diplostomidae) infectando olhos e cérebros de peixes na Amazônia Brasileira

The aim of this paper was to report the occurrence of Austrodiplostomum spp. in the eyes and brain of Acaronia nassa, Caquetaia spectabilis, Satanoperca acuticeps, Curimatella sp. and Crenicichla marmorata in a lake of the Amazon River system in the state of Pará (Brazil). Of 49 fish examined, 10.2% were parasitized by Austrodiplostomum spp. metacercariae, and A. nassa was the host with higher number of metacercariae. In five examined fish species, a total of 51 metacercariae were collected, with 45 found in the eyes and 6 in the cranial vault. In the eyes, the metacercariae were free and active in the vitreous humor, but no opacification was observed. In the brain, the metacercariae were also free and active, and located mainly below the encephalon, on the cranial floor, at the height of the ophthalmic lobes and near the optic nerve. In laboratory observations, however, the infected fish did not exhibit any behavioral disorders, and this may be related to the low level of parasitism. This was the first report of this digenean for C. spectabilis, S. acuticeps, Curimatella sp., C. marmorata and A. nassa.(AU)

O objetivo deste trabalho foi relatar a ocorrência de Austrodiplostomum spp. nos olhos e cérebros de Acaronia nassa, Caquetaia spectabilis, Satanoperca acuticeps, Curimatella sp. e Crenicichla marmorata em um lago do rio Amazonas no estado do Pará (Brasil). Dos 49 peixes examinados, 10,2% foram parasitados por metacercárias de Austrodiplostomum spp., A. nassa sendo o hospedeiro com maior número de metacercárias. Em cinco espécies de peixes examinadas, foram coletadas 51 metacercárias, sendo 45 encontradas nos olhos e 6 na caixa craniana. Nos olhos, as metacercárias estavam livres e ativas no humor vítreo, mas nenhuma opacificação foi observada. No cérebro, as metacercárias também eram livres e ativas, localizadas principalmente abaixo do encéfalo, no assoalho do crânio, na altura dos lobos oftálmicos e próximo ao nervo óptico. Em observações laboratoriais, no entanto, o peixe infectado não apresentou distúrbios comportamentais, o que pode estar relacionado ao baixo nível de parasitismo. Este foi o primeiro relato de Austrodiplostomum spp. em C. spectabilis, S. acuticeps, Curimatella sp., C. marmorata e A. nassa.(AU)

Echinostoma macrorchis Metacercariae in Cipangopaludina chinensis malleata from Xiengkhuang Province, Lao PDR and Morphologies of Adults from Experimental Animals

We identified the echinostome metacercariae in Chinese mystery snails, Cipangopaludina chinensis malleata, from Xiengkhuang Province, Lao PDR with morphologies of adult worms recovered. Total 20 snails were examined with artificial digestion method and then the collected metacercariae were orally infected to a mouse and a rat. Adult worms recovered from experimental animals were observed with a light microscope and a SEM. The metacercariae were round, 125×123 μm in average size, with a moderately thick cyst wall, collar spines distributed in the head collar and excretory granules in 2 canals of excretory tube. Adult flukes (3-week-old in a rat) were elongated, ventrally curved and 5.310×1.023 mm in average size. Head collar distinct, bearing 43 collar spines with 5 end group ones on each side. Oral sucker subterminal, prepharynx very short, pharynx well developed, and esophagus relatively short. Cirrus sac well developed, with a saccular seminal vesicle, and ventral sucker very large. Ovary round and on the median line of the body. Testes tandom and elongated. Eggs operculated, elliptical and 90×57 μm in average size. In the SEM observation, the head crown prominent, with 43 collar spines resembled with horns of younger stag. Scale-like tegumental spines were densely distributed on the surface between the head collar and ventral sucker, and their densities were decreased posteriorly. Conclusively, the metacercariae detected in C. chinensis malleata from Lao PDR were identified as those of Echinostoma macrorchis based on the morphological characteristics of adult worms.

Partial Characterization of Two Cathepsin D Family Aspartic Peptidases of Clonorchis sinensis

Cathepsin D (CatD, EC 3.4.23.5) is a member belonging to the subfamily of aspartic endopeptidases, which are classified into the MEROPS clan AA, family A1. Helminth parasites express a large set of different peptidases that play pivotal roles in parasite biology and pathophysiology. However, CatD is less well known than the other classes of peptidases in terms of biochemical properties and biological functions. In this study, we identified 2 novel CatDs (CsCatD1 and CsCatD2) of Clonorchis sinensis and partially characterized their properties. Both CsCatDs represent typical enzymes sharing amino acid residues and motifs that are tightly conserved in the CatD superfamily of proteins. Both CsCatDs showed similar patterns of expression in different developmental stages of C. sinensis, but CsCatD2 was also expressed in metacercariae. CsCatD2 was mainly expressed in the intestines and eggs of C. sinensis. Sera obtained from rats experimentally infected with C. sinensis reacted with recombinant CsCatD2 beginning 2 weeks after infection and the antibody titers were gradually increased by maturation of the parasite. Structural analysis of CsCatD2 revealed a bilobed enzyme structure consisting of 2 antiparallel β-sheet domains packed against each other forming a homodimeric structure. These results suggested a plausible biological role of CsCatD2 in the nutrition and reproduction of parasite and its potential utility as a serodiagnostic antigen in clonorchiasis.

Infection Status with Digenetic Trematode Metacercariae in Fishes from Coastal Lakes in Gangwon-do, Republic of Korea

The infection status of digenetic trematode metacercariae (DTM) was investigated in fishes from coastal lakes in Gangwon-do, the Republic of Korea (Korea). All fishes collected in 5 lakes were examined with the artificial digestion method. More than 10 species, i.e., Metagonimus spp., Pygidiopsis summa, Centrocestus armatus, Metorchis orientalis, M. taiwanensis, Clinostomum complanatum, Echinostoma spp., Stictodora spp., Diplostomum sp. and Diplostomid No. 1. by Morita (1960), of DTM were detected in fishes from 5 coastal lakes in Gangwon-do. Metagonimus spp. metacercariae were found in 52 (41.3%) out of 126 sea rundace, Tribolodon hakonensis, from 5 lakes, and their density was 14.6 per fish infected. P. summa metacercariae were detected in 48 (84.2%) out of 57 mullets from 5 lakes, and their density was 316 per fish infected. C. armatus metacercariae were detected in 7 (14.6%) T. hakonensis and 3 (15.0%) Tridentiger brevispinis from Hyang-ho, and 5 (19.2%) Acanthogobius flavimanus from Gyeongpo-ho. Stictodora spp. metacercariae were found in 4 fish species, i.e., Tridentiger obscurus, Tridentiger trigonocephalus, Chelon haematocheilus, and Acanthogobius lactipes, from Gyeongpo-ho. Total 15 C. complanatum metacercariae were detected in 2 (9.1%) crucian carp, Carassius auratus, from Songji-ho. M. taiwanensis metacercariae were found in T. hakonensis from Hyang-ho and Gyeongpo-ho and in Pseudorasbora parva from Gyeongpo-ho. Total 11 M. orientalis metacercariae were detected in 3 (6.3%) T. hakonensis from Hyang-ho. From the above results, it was confirmed that various species of DTM are infected in fishes from coastal lakes in Gangwon-do, Korea.

Survey of Gymnophalloides seoi Metacercariae in Natural and Cultured Oysters from Several Western Coastal Areas, Korea

Gymnophalloides seoi (Digenea: Gymnophallidae) is a human intestinal trematode contracted by eating raw oysters (Crassostrea gigas) in the Republic of Korea (=Korea). It has been known to be highly endemic in Aphae Island, Shinan-gun, Jeollanam-do (Province). However, recent epidemiological status of G. seoi has not been reported since the 1990s. In this study, we investigated the prevalence of G. seoi metacercariae in natural and cultured oysters collected from 3 islands and 2 coastal areas in western parts of Korea. The oysters were examined using the artificial digestion method followed by stereomicroscopy. The overall positive rate of G. seoi metacercariae in natural oysters was 66.0% (99/150), and the oysters collected from Yubu Island showed the highest infection rate (74.0%). However, the metacercarial density per oyster was relatively low (1.5–2.4 per oyster). By contrast, no metacercaria was found in cultured oysters purchased from 2 coastal areas in Chungcheongnam-do. Thus, we could confirm that natural oysters produced from 3 western coastal islands are infected with G. seoi metacercariae, whereas cultured oysters purchased from 2 coastal areas were free from infection.

Infection Status with Clinostomum complanatum Metacercariae in Fish from Water Systems of Nakdong-gang (River) in Korea

The infection status of Clinostomum complanatum metacercariae (CcMc) was broadly surveyed in fishes from water systems of Nakdong-gang (River) in the Republic of Korea (Korea) for 5 years. All 4,468 fishes collected were individually examined by the artificial digestion methods. No CcMc were detected in fishes from Naeseongcheon in Yecheon-gun, Gigyecheon in Gyeongju-si, Gyeongsangbuk-do, and Hamancheon in Haman-gun, Gyeongsangnam-do. In fishes from Wicheon in Gunwi-gun, Gyeongsangbuk-do, CcMc were detected in 180 (15.4%) out of 1,168 fishes and their intensity was 5.8 per fish infected. The prevalences with CcMc in positive fish species from 3 other surveyed areas, i.e., Banbyeoncheon in Yeongyang-gun, Nakdong-gang in Sangju-si and Hoecheon in Goryeong-gun, in Gyeongsangbuk-do were 19.3%, 33.3%, and 19.0% and their intensities were 1.5, 17.6, and 2.6 per fish infected respectively. In fishes from Yangcheon in Sancheong-gun, Gyeongsangnam-do, CcMc were detected in 811 (45.6%) out of 1,779 fishes examined and their intensity was 9.8 per fish infected. The prevalence was most high in Squalidus spp. (97.7%) and followed by acheilognathinid fish (66.8%), Pungtungia herzi (52.0%), rasborinid fish (39.6%) and Hemibarbus spp. (25.9%) from Yangcheon. The intensity was also most high in Squalidus spp. (27.0). The endemicity with CcMc was very low in fishes from water systems of Nakdong-gang in the east coast of Korea. Conclusively, it is confirmed that various fish species act as the second intermediate hosts of C. complanatum, and the endemicities of CcMc are more or less different by the fish species and group from Yangcheon, in Sancheong-gun, Gyeongsangnam-do, Korea.

Morphological and Molecular Identification of Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae) in Thailand

We tried a series of morphological and molecular approaches to identify a new species of Stellantchasmus (Digenea: Heterophyidae) originating from the wrestling half-beaked fish, Dermogenys pusillus of Thailand. Adult worm samples of the new species were recovered from hamsters experimentally infected with the metacercariae from D. pusillus in Thailand. Two isolates (Thai and Korean) of Stellantchasmus falcatus were used as comparative control groups. Worm samples of 3 Stellantchasmus groups were morphologically observed and molecularly analyzed with the mitochondrial cytochrome c oxidase 1 gene. The morphological characteristics of S. dermogenysi n. sp. are similar to S. falcatus originating from brackish water fish, but minor difference was noted including the absence of the prepharynx, position of the ovary near the ceca end, smaller body size, and shorter esophageal length. A phylogenetic tree derived from neighbor-joining and maximum-likelihood methods suggests that S. dermogenysi n. sp. is separated from S. falcatus supported by high bootstrap values. The relative divergences persist between these host-specific trematodes, which we suggest should be recognized as 2 distinct species. Comparisons of S. dermogenysi n. sp. with S. falcatus isolated from mullets in Thailand and Korea indicate a genetic divergence of mitochondrial DNA of 19.4% and 21.7%, respectively. By the present study, a new species, Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae), is proposed in Thailand based on molecular evidences, in addition to minor morphological differences between S. falcatus and the new species.

Infections with Digenetic Trematode Metacercariae in Freshwater Fishes from Two Visiting Sites of Migratory Birds in Gyeongsangnam-do, Republic of Korea

The infection status of digenetic trematode metacercariae (DTM) was investigated in fishes from 2 representative visiting sites of migratory birds in Gyeongsangnam-do, the Republic of Korea (Korea). A totaly 220 freshwater fishes (7 species) were collected from Junam-jeosuji (reservoir), and 127 fishes (7 species) were also collected from Woopo-neup (swamp) in June and October 2017. As the control group, total 312 fish (22 spp.) from Yangcheon in Sancheong-gun, Gyeongsangnam-do were also collected in June and October 2017. All fishes collected in 3 sites were examined with the artificial digestion method. In the fishes from Junam-jeosuji, more than 4 species, i.e., Clonorchis sinensis, Echinostoma spp., Diplostomum spp. and Cyathocotyle orientalis, of DTM were detected and their endemicy was very low, 0.70. More than 6 species, i.e., C. sinensis, Echinostoma spp., Metorchis orientalis, Clinostomum complanatum, Diplostomum spp. and C. orientalis, of DTM were found in the fishes from Woopo-neup, and their endemicy was low, 5.16. In the fishes from Yangcheon, more than 8 species, i.e., C. sinensis, Metagonimus spp., Centrocestus armatus, C. complanatum, C. orientalis, M. orientalis, Echinostoma spp., and Diplostomum spp., of DTM were detected, and their endemicity was relatively high, 95.48. The percentages of avian trematode metacercariae (ATM) were 99.6% and 94.7% in fishes from Junam-jeosuji and Woopo-neup whereas it was 74.1% in the control site, Yangcheon. The above findings suggested that migratory birds partly affect in endemicity of DTM in the fish in the 2 visiting sites in Gyeongsangnam-do, Korea.

Infection Status with Clonorchis sinensis Metacercariae in Fish from Yangcheon (Stream) in Sancheong-gun, Gyeongsangnam-do, Korea

The infection status with Clonorchis sinensis metacercariae (CsMc) was examined in freshwater fishes from Yangcheon (a branch of Gyeongho-gang), which is located in Sancheong-gun, Gyeongsangnam-do, the Republic of Korea. Total 2,201 fishes in 26 species were examined by the artificial digestion method through 7 years. CsMc were detected in 1,171 (53.2%) fishes in 21 spp. (80.8%) and their density was 85 per fish infected. Total 532 (99.6%) out of 534 Pungtungia herzi (index fish) examined were infected with 147 CsMc per fish infected. Metacercarial densities in this fish were highest in 2015 (179 CsMc), followed by 2012 (168), 2013 (152), 2016 (145), 2014 (114), and 2017 (89) respectively. In the gobioninid fish group, i.e., P. herzi, Sarcocheilichthys spp., Squalidus spp., Pseudogobio esocinus, Hemibarbus longirostris, and Hemibarbus labeo, 841 (92.7%) fishes were infected with 117 CsMc per fish infected. Total 250 (54.7%) acheilognathinid fish (bitterlings), Acheilognathus spp. and Acanthorhodeus spp. were infected with 5.8 CsMc. In the rasborinid fish (chubs) group, i.e., Zacco platypus, Zacco temminckii, and Zacco koreanus, 77 (13.7%) out of 563 fish examined were infected with 2.4 CsMc in average. The susceptibility indices of CsMc were 49.09 in the overall positive fish group, 104.15 in the gobioninid group, 3.17 in the acheilognathinid group and 0.35 in the rasborinid fish group respectively. Only 1 CsMc was detected in 3 fish species, Coreoperca herzi, Channa argus, and Lepomis macrochirus, respectively. Conclusively, it was confirmed that CsMc are moderately prevalent in fishes from Yangcheon in Sancheon-gun, Gyeongsangnam-do, Korea.

Metacercariae of Tylodelphys sp. (Trematoda: Diplostomidae) parasite of Brochis multiradiatus and Corydoras splendens (Siluriformes: Callichthyidae) from the Peruvian Amazon / Metacercárias de Tylodelphys sp. (Trematoda: Diplostomidae) parasito de Brochis multiradiatus e Corydoras splendens (Siluriformes: Callichthyidae) da Amazônia peruana

Corydoras splendens (Castelnau, 1855) and Brochis multiradiatus (Orcés V., 1960) are highly demanded for exporters in the Peruvian Amazon and importers all over the world. The exportation of this fish species may represent a gateway for endoparasites to enter in a new environment with serious effects, so, the present study reports the presence of Tylodelphys sp. infecting C. splendens and B. multiradiatus. Fish were acquired from fishermen of the city of Iquitos, Peru. After the necropsy and analyses of the fish, metacercariae of Tylodelphys sp. were found in the liver and intestine. This finding represents the first report of Tylodelphys Diesing, 1850 infecting a fish in the Peruvian Amazon. Considering the possible damages caused by species of Tylodelphys, highlighting the nutritional and respiratory ones, prophylactic measures must be taken into consideration for exporters to avoid some damages in the fish that can deteriorate their health and also to avoid the introduction of this parasite to other countries.

Corydoras splendens (Castelnau, 1855) e Brochis multiradiatus (Orcés V., 1960) são altamente demandados por exportadores da Amazônia peruana e importadores em todo o mundo. A exportação dessas espécies de peixes pode representar uma entrada de endoparasitas a um novo ambiente com efeitos graves. Assim, o presente estudo relata a presença de Tylodelphys sp. infectando C. splendens e B. multiradiatus. Os peixes foram adquiridos de pescadores da cidade de Iquitos, Peru. Após a necropsia e análise dos peixes, metacercarias de Tylodelphys sp. foram encontrados no fígado e no intestino, representando o primeiro registro de Tylodelphys Diesing, 1850 infectando um peixe na Amazônia peruana. Considerando os possíveis danos causados por espécies de Tylodelphys, destacando-se os danos nutricionais e respiratórios; medidas profiláticas devem ser levadas em consideração pelos exportadores para evitar alguns danos nos peixes que podem comprometer a saúde e também a introdução deste parasita em outros países.

Infection Status of Isthmiophora hortensis Metacercariae in Dark Sleepers, Odontobutis Species, from Some Water Systems of the Republic of Korea

Present study was performed to survey on infection status of Isthmiophora hortensis (formerly Echinostoma hortense) metacercariae (IhMc) in dark sleepers, Odontobutis spp., from some water systems of the Republic of Korea. A total of 237 Odontobutis spp. was collected in the water systems of 5 rivers, i.e., Mangyeong-gang (gang means river), Geum- gang, Tamjin-gang, Seomjin-gang, and Nakdong-gang. They were all examined with artificial digestion method for 5 years (2013–2017). A total of 137 (57.8%) Odontobutis spp. were infected with 14.8 IhMc in average. The prevalence was the highest in Nakdong-gang areas (62.9%) and followed by in Mangyeong-gang (57.1%), Geum-gang (56.3%), Tamjingang (54.8%), and Seomjin-gang (53.9%) areas. Metacercarial densities were 28.1 (Geum-gang), 13.9 (Mangyeong-gang), 13.3 (Nakdong-gang), 13.1 (Tamjin-gang), and 2.3 (Seomjin-gang) per infected fish. Especially, in case of Yugucheon (cheon means stream), a branch of Geum-gang, IhMc were detected in all fish (100%) examined and their density was about 48 per fish. By the present study, it was confirmed that the infection status of IhMc is more or less different by the surveyed areas and the dark sleepers, Odontobutis spp., are suitable fish hosts of I. hortensis.

Morphological Characteristics and Fish Hosts of Isoparorchis sp. (Digenea: Isoparorchiidae) in Korea

We described here the morphological characteristics for the species identification and fish hosts of Isoparorchis sp. (Digenea: Isoparorchiidae) in the Republic of Korea (Korea). Total 1,371 freshwater fishes collected in Yangcheon (Stream) in Sancheong-gun, Gyeongsangnam-do were examined by the artificial digestion methods to survey the infection status of digenetic trematode metacercariae for 4 years (2013–2016). Adult and larval worms of Isoparorchis sp. were detected in 38 (8.4%) out of 451 fish in 4 species, i.e., Pungtungia herzi, Acheilognathus koreensis, Squalidus japonicus coreanus and Odontobutis platycephala, examined. The infection density was 1.1 worm per fish infected. They were mainly found in the subcutaneous tissues and abdominal cavities. Nodules with worms in the subcutaneous tissues were revealed as the blue ink-colored bulges. Adults leaf-like, 21.6×9.84 mm in average size. The ratio of body length to body width was 2.20: 1. Oral sucker subterminal, 1.03×1.22 mm. Pharynx muscular, 0.55×0.54 mm. Esophagus very short. Ceca convoluted, terminated near the posterior end. Ventral sucker anterior 1/3.75, 1.99×2.10 mm. The ratio of ventral sucker to oral sucker was 1.74: 1. Testes round to elliptical, both sides of ventral sucker, 1.43×1.33 mm. Vitellaria highly dendritic, posterior 1/3 level. Eggs operculated, embryonated, 52×32 μm in size. By the present study, 4 fish species aforementioned are to be listed as the fish hosts of Isoparorchis sp. in Korea and additionally the morphological characteristics are to be described for the species identification.

Infections with Digenean Trematode Metacercariae in Two Invasive Alien Fish, Micropterus salmoides and Lepomis macrochirus, in Two Rivers in Chungcheongbuk-do, Republic of Korea

Present study was performed to survey infection status of digenetic trematode metacercariae in 2 alien fish species, Micropterus salmoides (largemouth bass) and Lepomis macrochirus (bluegill), in 2 rivers draining Chungcheongbuk-do, Republic of Korea. A total of 107 largemouth bass and 244 bluegills were caught in Daecheong-ho (ho=lake) and Musim-cheon (a branch of Geum-gang), in Chungcheongbuk-do April–July 2015. Additionally, 68 native fish of 5 species, i.e., Zacco platypus, Hemibarbus longirostris, Carassius auratus, Pseudogobio esocinus and Puntungia herzi, were caught from the same water bodies. All of the fish collected were examined by artificial digestion method. The metacercariae of Centrocestus armatus, Clinostomum complanatum, Metagonimus sp. and Diplostomum spp. were detected from 4 out of 5 native fish species in Daecheong-ho. However, any metacercariae were not found from 87 M. salmoides and 177 L. macrochirus in Daecheong-ho. In Musim-cheon, metacercariae of Exorchis oviformis and Metacercaria hasegawai were detected from 78% Z. platypus and 34% L. macrochirus, but any metacercariae not found in M. salmoides. We report here that the 2 alien fish species were less infected with the metacercariae than the native ones. Surveys on the metacercariae in the alien fish species in geographically various rivers should be undertaken for better understanding on the role of alien fish species in the trematode infections in Republic of Korea.

Infections with Centrocestus armatus Metacercariae in Fishes from Water Systems of Major Rivers in Republic of Korea

The infection status of Centrocestus armatus metacercariae (CaMc) was broadly surveyed in freshwater fishes from major river systems in the Republic of Korea (Korea) during 2008–2017. A total of 14,977 fishes was caught and examined by the artificial digestion method. CaMc were detected in 3,818 (97.1%) (2,114 Z. platypus: 96.1% and 1,704 Z. temminckii: 98.4%) out of 3,932 Zacco spp. examined and their density was 1,867 (2,109 in Z. platypus and 1,567 in Z. temminckii) per fish infected. The prevalences with CaMc were high, 93.7–100%, in Zacco spp. from all surveyed areas. However, their densities were more or less different by the surveyed areas and fish species. They were most high in Nakdong-gang in Gyeongsangnam-do (4,201 in average), and followed by Geum-gang (2,343), Nakdong-gang in Gyeongsangbuk-do (1,623), Han-gang (1,564), Tamjin-gang and Yeongsan-gang (1,540), streams in the east coast (1,028), Seomjin-gang (488) and Mangyeong-gang (170). In another species of rasborinid fish, Opsariichthys uncirostris amurensis, CaMc were detected in 222 (74.8%) out of 297 ones examined and their density was 278 (1–4,480) per fish infected. CaMc were also detected in total 41 fish species except for the rasborinid fish, Z. platypus, Z. temminckii and O. uncirostris amurensis. Conclusively, it was confirmed that among the 3 species of rasborinid fish, Z. platypus and Z. temminckii are highly prevalent and O. uncirostris amurensis is moderately prevalent with CaMc. Additionally, we could know that variety of fish species act as the second intermediate hosts of C. armatus in Korea.

Infection Status with Metagonimus spp. Metacercariae in Fishes from Seomjin-gang and Tamjin-gang in Republic of Korea

To grasp the infection status of Metagonimus spp. metacercariae (MsMc), the freshwater fishes were surveyed from Seomjin-gang (river) and Tamjin-gang in the Republic of Korea. Total 1,604 fishes from 7 local sites of Seomjin-gang and 1,649 fishes from 2 sites of Tamjin-gang were examined for 6 years (2012–2017) by the artificial digestion method. MsMc were detected in fishes from 7 sites, i.e., Osucheon in Imsil-gun (36.3% fish in 6 spp.), Seomjin-gang in Sunchanggun (49.8% in 18 spp.), Songdaecheon in Namwon-si (64.5% in 8 spp.), Seomjin-gang in Gokseong-gun (72.4% in 14 spp.) and in Gurye-gun (78.8% in 17 spp.), Hoengcheon (75.9% in 11 spp.) and Namsancheon (58.9% in 7 spp.) in Hadong-gun. Their average densities were 4.2, 86.8, 39.9, 43.1, 246.5, 173.6 and 67.5 per fish infected respectively. Prevalence with MsMc in rasborinid fish from Seomjin-gang was 89.2% and their intensity was 73.6 per fish infected. Endemicities of MsMc in rasborinid fish from Seomjin-gang were significantly higher in fishes from the lower reaches (prevalence: 98.7%; density: 137) rather than in fishes from the middle (93.5%; 38) and upper (72.4%; 13) reaches. MsMc were also detected in 56.1% and 66.4% fishes from 2 surveyed areas, i.e., the middle reaches in Jangheung-gun and the lower reaches in Gangjin-gun, of Tamjin-gang, and their densities were 147 and 121 per fish infected. In susceptible fishes from Tamjin-gang, the prevalence was 84.1% and density was 227 per fish infected. By the present study, it was confirmed that MsMc is highly prevalent in the fishes from Seomjin-gang and Tamjin-gang in Korea.

Freshwater Snail Diversity in Mae Lao Agricultural Basin (Chiang Rai, Thailand) with a Focus on Larval Trematode Infections

The aim of this study was to conduct a freshwater snail survey in Mae Lao agricultural basin to assess the diversity with a focus on habitat types and their larval trematode infections. Snails were collected and examined in 14 sites of Mae Lao agricultural basin from August 2016 to October 2017. A total of 1,688 snail individuals were collected and classified into 7 families, 8 genera, and 12 species. Snail diversity and habitat types were higher in rice paddies than irrigation canals and streams. The most abundant species was Bithynia siamensis siamensis, representing 54.6% of the sample. Three species of snails act as first intermediate host were found with cercarial infections. They were Filopaludina sumatrensis polygramma, B. s. siamensis, and Melanoides tuberculata. The cercariae were categorized into 7 types; echinostome, monostome, gymnocephalous, virgulate, parapleurolophocercous, pleurolophocercous and megalurous cercariae. Parapleurolophocercous cercariae constituted the most common type of cercariae recovered, contributing 41.2% of all infections in snails. Echinostome metacercariae infections were found in 6 snail species with 7.6% prevalence. In addition, the metacercaria of avian trematode, Thapariella sp. were found in Filopaludina spp. snails and B. funiculata with a prevalence of 0.5%. This is the first report for Thapariella metacercariae in the snail host, B. funiculata, and also confirmed that viviparid and bithyniid snails act as the second intermediate hosts of this trematode. This work will provide new information on the distribution and intermediate host of trematode in this area.