First report of neurotoxic effect of the cyanobacterium Cylindrospermopsis raciborskii on the motility of trematode metacercariae.

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Lopes, K C; Ferrão-Filho, A S; Santos, E G N; Santos, C P

2018-03-01

Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.

[Identification of Clonorchis sinensis metacercariae based on PCR targeting ribosomal DNA ITS regions and COX1 gene].

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Yang, Qing-Li; Shen, Ji-Qing; Jiang, Zhi-Hua; Yang, Yi-Chao; Li, Hong-Mei; Chen, Ying-Dan; Zhou, Xiao-Nong

2014-06-01

To identify Clonorchis sinensis metacercariae using PCR targeting ribosomal DNA ITS region and COX1 gene. Pseudorasbora parva were collected from Hengxian County of Guangxi at the end of May 2013. Single metacercaria of C. sinensis and other trematodes were separated from muscle tissue of P. parva by digestion method. Primers targeting ribosomal DNA ITS region and COX1 gene of C. sinensis were designed for PCR and the universal primers were used as control. The sensitivity and specificity of the PCR detection were analyzed. C. sinensis metacercariae at different stages were identified by PCR. DNA from single C. sinensis metacercaria was detected by PCR targeting ribosomal DNA ITS region and COX1 gene. The specific amplicans have sizes of 437/549, 156/249 and 195/166 bp, respectively. The ratio of the two positive numbers in PCR with universal primers and specific primers targeting C. sinensis ribosomal DNA ITS1 and ITS2 regions was 0.905 and 0.952, respectively. The target gene fragments were amplified by PCR using COX1 gene-specific primers. The PCR with specific primers did not show any non-specific amplification. However, the PCR with universal primers targeting ribosomal DNA ITS regions performed serious non-specific amplification. C. sinensis metacercariae at different stages are identified by morphological observation and PCR method. Species-specific primers targeting ribosomal DNA ITS region show higher sensitivity and specificity than the universal primers. PCR targeting COX1 gene shows similar sensitivity and specificity to PCR with specific primers targeting ribosomal DNA ITS regions.

Host records and tissue locations for Diplostomum mordax (metacercariae) inhabiting the cranial cavity of fishes from Lake Titicaca, Peru.

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Heckmann, R A

1992-06-01

Metacercariae of Diplostomum mordax were found in the cranial cavity of Orestias agasii, Orestias olivaceous, Orestias luteus, and Basilichthys bonariensis, fishes from Lake Titicaca, Peru. Metacercariae were not found in Oncorhynchus mykiss introduced into the lake during 1939 and 1940. Compression of neural tissue within and on the surface of the brain was observed in all infected fishes. Metacercariae migrating into the cerebrum and cerebellum of the piscine host caused hemorrhaging, cell necrosis, inflammation, fiber formation, and nerve fiber disruption. The presence of D. mordax in B. bonariensis and the 3 species of Orestias constitute new host records. Infections in the cerebrum and cerebellum add new information on specific parasite location.

La infección de cangrejos procedentes del valle de Condebamba (Cajamarca por metacercarias de Paragonimus

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Manuel Tantaleán V.

2014-06-01

Full Text Available En los meses de febrero y julio de 1973 se colectaron cangrejos del Valle de Condebamba (Cajamarca correspondientes a Pseudothelphusa chilensis, 69 de ellos (34 machos y 35 hembras fueron examinados, buscando metacercarias de Paragonimus; 22 (64.7% de los machos y 29 (28.85 % de las hembras resultaron infectados con metacercarias, o sea 52 (72.46% de los 69 cangrejos estaban infectados, esta cifra es coincidente con la obtenida por Cuba y cols. (1974.

Paragonimus y paragonimiasis en el norte peruano. Infección natural de Pseudothelphusa chilensis por metacercarias de Paragonimus Braun, 1899

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César Cuba

2014-06-01

Full Text Available Se ha estudiado el índice de infección natural y la intensidad del parasitismo en cangrejos recolectados en tres provincias del Departamento de Cajamarca, Perú, habiéndose encontrado metacercarias de Paragonimus Braun, 1899 en el 43.97% de 539 especímenes de Pseudothelphusa chilensis Milne Edwards, 1843, único cangrejo hallado en la zona. El órgano parasitado fue casi exclusivamente el hepatopancreas, siendo 10.33 el número promedio de metacercarias por cangrejo. Las formas adultos logradas mediante inoculación de las metacercarias, fueron identificadas como Paragonimus peruvianus Miyazaki, Ibáñez y Miranda, 1969, con excepción de tres ejemplares que correspondieron a lo especie Paragonimus caliensis Little, 1968.

Prevalence of Clonorchis sinensis Metacercariae in Fish from Water Systems of Seomjin-gang (River).

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Sohn, Woon-Mok; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Kim, Cheon-Hyeon; Hwang, Min-Ah; No, Kyeong-Woo; Yoon, Ki-Bok; Lim, Hyun-Cheol

2017-06-01

The prevalence of Clonorchis sinensis metacercariae ( Cs Mc) was examined in freshwater fish from the water systems of Seomjin-gang (River), the Republic of Korea. Total 1,604 fish from 7 local sites of Seomjin-gang were examined by artificial digestion methods. The metacercariae of C. sinensis were detected in 102 (39.8%) out of 256 fish (14 species) from the upper reaches of Seomjin-gang, i.e., Osucheon (22.3% in 6 fish species) in Imsil-gun, and Seomjin-gang (63.9% in 9 fish species) in Sunchang-gun, Jeollabuk-do. Their average density was 9.0 per infected fish. They were also found in 132 (48.0%) out of 275 fish (12 spp.) from the middle reaches of Seomjin-gang, i.e., Songdaecheon (58.9% in 4 fish species) in Namwon-si, Jeollabuk-do, and Seomjin-gang (45.2% in 10 fish species) in Gokseong-gun, Jeollanam-do. Their average density was 21.0 per infected fish. Cs Mc were detected in 77 (56.6%) out of 136 fish (11 species) from the lower reaches of Seomjin-gang, i.e., Seomjin-gang (73.3% in 11 fish species) in Gurye-gun, Jeollanam-do, and Namsancheon (8.6% in 1 fish species) in Hadong-gun, Gyeongsangnam-do. Their average density was 64.9 per infected fish. The metacercariae of Metorchis orientalis were also detected in 6 fish species from 4 sites of Seomjin-gang. Conclusively, it has been confirmed that Cs Mc are more or less prevalent in fish from some water systems of Seomjin-gang in Korea.

Prevalence of Centrocestus formosanus Metacercariae in Ornamental Fish from Chiang Mai, Thailand, with Molecular Approach Using ITS2.

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Wanlop, Atcharaphan; Wongsawad, Chalobol; Prattapong, Pongphol; Wongsawad, Pheravut; Chontananarth, Thapana; Chai, Jong-Yil

2017-08-01

The prevalence of Centrocestus formosanus metacercariae was investigated in ornamental fish purchased from a pet shop in Chiang Mai, Thailand, including Carassius auratus (goldfish), Cyprinus carpio (Koi), Poecilia latipinna (Sailfin Molly), Danio rerio (Zebrafish), and Puntigrus tetrazona (Tiger barb). The parasite species was identified by the morphology of worms as well as by a molecular approach using ITS2. The results showed that 50 (33.3%) of 150 fish examined were infected with the metacercariae. The highest prevalence was found in C. auratus (83.3%), and the highest intensity was noted in C. carpio (70.8 metacercariae/fish). The most important morphological character was the presence of 32-34 circumoral spines on the oral sucker. The phylogenetic studies using the rRNA ITS2 region revealed that all the specimens of C. formosanus in this study were grouped together with C. formosanus in GenBank database. This is the first report on ornamental fish, C. carpio, P. latipinna, D. rerio, and P. tetrazona, taking the role of second intermediate hosts of C. formosanus in Thailand. Prevention and control of metacercarial infection in ornamental fish is urgently needed.

Effect of citric acid on the acidification of artificial pepsin solution for metacercariae isolation from fish.

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Kim, Min-Ki; Pyo, Kyoung-Ho; Hwang, Young-Sang; Chun, Hyang Sook; Park, Ki Hwan; Ko, Seong-Hee; Chai, Jong-Yil; Shin, Eun-Hee

2013-11-15

Artificial digestive solution based on pepsin is essential for collecting metacercariae from fish. To promote the enzymatic reactivity of pepsin, the pH of the solution has to be adjusted to pH 1.0-2.0. Hydrochloride (HCl) is usually used for this purpose, but the use of HCl raises safety concerns. The aim of this work was to address the usefulness of citric acid as an alternative for HCl for the acidification of pepsin solution, and to examine its potential to damage metacercariae during in vitro digestion as compared with HCl. Changes in pH after adding 1-9% of citric acid (m/v) to pepsin solution were compared to a 1% HCl (v/v) addition. Digestion of fish muscle was evaluated by measuring released protein concentrations by spectrophotometry. In addition, survival rates of metacercariae in pepsin solution were determined at different citric acid concentrations and were compared that of with 1% HCl. The present study shows that addition of citric acid reduced the pH of pepsin solutions to the required level. Addition of more than 5% of citric acid resulted in the effective digestion of fish muscle over 3h in vitro, and 5% citric acid was less lethal to metacercariae than 1% HCl in pepsin solution. Pepsin solution containing 5% citric acid had digestive capacity superior to pepsin solution containing 1% HCl after 3h incubation with released protein concentrations of 12.0 ng/ml for 5% citric acid and 9.6 ng/ml for 1% HCl. Accordingly, the present study suggests that the addition of 5% citric acid to pepsin solution is a good alternative to 1% HCl in infection studies because citric acid is a stable at room temperature and has a good safety profile. In addition, we suggest that the use of citric acid enables the preparation of commercial digestive solutions for the detection of microorganisms in fish and other vertebrate muscle tissue. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

In vitro cultivation of Cymatocarpus solearis (Brachycoeliidae) metacercariae to obtain the adult stage without the marine turtle definitive host.

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Grano-Maldonado, Mayra; Alvarez-Cadena, José

2010-03-01

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at 38 to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adult-stage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n = 87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis.

In Vitro Cultivation of Cymatocarpus solearis (Brachycoeliidae) Metacercariae to Obtain the Adult Stage without the Marine Turtle Definitive Host

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Álvarez-Cadena, José

2010-01-01

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at 38℃ to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adult-stage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n = 87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis. PMID:20333285

Distribución parcial de Paragonimus (Digenea: Troglotrematidae en Antioquia, por presencia de metacercarias en cangrejos dulciacuícolas

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Luz Elena Velásquez

2008-12-01

Full Text Available Introducción. La paragonimosis, o distomatosis pulmonar, es una enfermedad con sintomatología similar a la observada en la tuberculosis. Es causada por parásitos del género Paragonimus (Digenea: Troglotrematidae. Las personas se infectan al consumir cangrejos crudos o mal cocidos, con metacercarias del parásito. El primer foco de paragonimosis humana en Colombia se registró durante 1995 en Urrao, Antioquia, donde se hallaron dos especies de cangrejos que hospedaban el parásito. En el 2005 se capturaron cangrejos con metacercarias de Paragonimus en Medellín, lo que motivó la búsqueda del parásito en otras localidades, mediante su presencia en estos crustáceos. Objetivo. Establecer la distribución de Paragonimus en Antioquia, evaluando la presencia de metacercarias en macrocrustáceos braquiuros, dulciacuícolas. Materiales y métodos. Desde 2005 hasta 2007 se capturaron cangrejos en 13 municipios antioqueños. Se relajaron y sacrificaron para la búsqueda del digeneo y la identificación taxonómica. Resultados. En nueve municipios se capturaron 52 cangrejos, 42 (80,76% con metacercarias de Paragonimus. Todos los crustáceos se determinaron como Pseudothelphusidae, de los géneros Hypolobocera y Strengeriana, y se asignaron a cuatro especies. Tres se registran por primera vez como huéspedes del parásito. Conclusión. Se inicia la construcción de un mapa con la distribución de Paragonimus en Antioquia que incluye por primera vez zonas urbanizadas. Se ratifican el consumo y la manipulación de los cangrejos crudos y mal cocidos como factores de riesgo para la infección humana. Se propone a los cangrejos como agentes focalizadores de paragonimosis por ser asequibles y de fácil diagnóstico.

Morphological and molecular identification of the fish-borne metacercaria of Ascocotyle (Phagicola) longa Ransom, 1920 in Mugil liza from Argentina.

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Martorelli, S R; Lino, A; Marcotegui, P; Montes, M M; Alda, P; Panei, C J

2012-12-21

This is the first report of Ascocotyle (Phagicola) longa Ransom, 1920 (Digenea: Heterophyidae) in Argentina confirmed by morphological and molecular studies. The metacercaria was found encysted in myotomal musculature, heart and mesentery of the mullet Mugil liza (Pisces: Mugilidae) from Samborombon bay. We provide a morphological description of the metacercaria which we identified using species-specific primers for A. (Phagicola) longa and nucleotid sequence. This worldwide parasite has been reported as one of the causative agents of heterophyiosis, an emerging fish-borne disease of humans, contracted by the consumption of raw mullet. The discovery of A. (Phagicola) longa in Argentina represents a warning of the potentially great impact of this parasite on public health. Copyright © 2012 Elsevier B.V. All rights reserved.

Occurrence and effect of trematode metacercariae in two endangered killifishes from Greece.

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Kalogianni, Eleni; Kmentová, Nikol; Harris, Eileen; Zimmerman, Brian; Giakoumi, Sofia; Chatzinikolaou, Yorgos; Vanhove, Maarten P M

2017-11-01

We report digeneans (Diplostomidae, Crassiphialinae) in the endangered freshwater fishes Valencia letourneuxi and Valencia robertae, endemics of Western Greece. Digenean metacercariae occurred in two forms in the abdominal cavity, excysted and encysted, the latter attached to the gonads, liver and alimentary tract. Parasites were, using morphological and molecular techniques, identified as two representatives of Crassiphialinae, specifically part of the Posthodiplostomum-Ornithodiplostomum clade. The spatial, seasonal, and age class variation in parasite prevalence was examined. Autumn parasite prevalence varied between the six populations sampled (18.2 to 100%). Seasonal prevalence at the two sites sampled quadannually peaked in autumn and reached its lowest value in spring; prevalence increased with size to 100% in young adult fish. We did not find a correlation between prevalence and host sex. Overall parasites' weight averaged 0.64% of the host's, while parasite weight increased with host weight. A comparison of relative condition and hepatosomatic and gonadosomatic indices of infected and metacercariae-free specimens showed that infection did not have a significant effect on host body condition and reproduction. Regarding the parasite's life cycle, planorbid gastropods are proposed as potential first intermediate hosts in view of the host's diet and occurrence data of molluscs in the ecosystem. This is the first record of a diplostomid digenean in valenciid fishes and of representatives of the Posthodiplostomum-Ornithodiplostomum clade in a native Greek freshwater fish. Our findings are discussed in conjunction to fish conservation interventions, since parasites may contribute to the decline of endangered species.

Co-existence of Paragonimus harinasutai and Paragonimus bangkokensis metacercariae in fresh water crab hosts in central Viet Nam with special emphasis on their close phylogenetic relationship.

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Doanh, Pham Ngoc; Hien, Hoang Van; Nonaka, Nariaki; Horii, Yoichiro; Nawa, Yukifumi

2012-09-01

During our epidemiological surveys for Paragonimus species in central Viet Nam, we found four morphologically different Paragonimus metacercariae in mountainous crabs. They were identified as metacercariae of Paragonimus westermani, P. bangkokensis, P. proliferus, and P. harinasutai in the order of their prevalence in crab hosts. This is the first discovery of P. harinasutai in Viet Nam, co-inhabiting with P. bangkokensis and other species. Metacercariae of P. harinasutai were given orally to a cat to obtain adult worms. Then, ITS2 and CO1 sequences of metacercariae and adults of P. harinasutai, and metacercariae of P. bangkokensis collected from the same place were determined for analyses of phylogenetic relationships to other P. harinasutai and P. bangkokensis populations as well as related species. The results of molecular analyses showed that P. harinasutai from Quang Binh province of central Viet Nam was almost completely identical with those from Vientiane, Lao PDR; P. bangkokensis from Quang Binh, Viet Nam was also almost completely identical with those from Lao PDR and from Quang Ninh province, Viet Nam. Except for one P. harinasutai isolate from China, all populations of P. harinasutai and P. bangkokensis from Thailand, Lao and Viet Nam make a single clade in both ITS2 and CO1 trees. In ITS2 sequences, AT deletion and ATC insertion were observed in some isolates of both species, indicating recent gene flow between P. harinasutai and P. bangkokensis. Moreover, because of their extremely high genetic similarities and their co-inhabitation in the same crab hosts found in Thailand, Lao PDR and Viet Nam, they should be considered as the sister species at the early stage of divergence. In addition, P. microrchis previously described from Yunnan, China should be placed as the synonym of P. harinasutai, because of their morphological and molecular similarities. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Electrophoretic pattern of sera from lambs and kids vaccinated with irradiated Amphistome metacercariae (Cercariae indicae XXVI)

International Nuclear Information System (INIS)

Hafeez, Md.; Rao, B.V.

1986-01-01

Preliminary work has been done to study certain responses induced by irradiated amphistome metacercariae used as a vaccine to immunise lambs, kids and calves. The electrophoretic pattern of the sera collected from lambs and kids vaccinated with gamma irradiated amphistome matacercariae (C.I. XXVI) has been reported in this study. (author). 10 refs., 1 table

Identification and Characterization of Paramyosin from Cyst Wall of Metacercariae Implicated Protective Efficacy against Clonorchis sinensis Infection

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Lv, Xiaoli; Tian, Yanli; Men, Jingtao; Zhang, Xifeng; Lei, Huali; Zhou, Chenhui; Lu, Fangli; Liang, Chi; Hu, Xuchu; Xu, Jin; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing

2012-01-01

Human clonorchiasis has been increasingly prevalent in recent years and results in a threat to the public health in epidemic regions, motivating current strategies of vaccines to combat Clonorchis sinensis (C. sinensis). In this study, we identified C. sinensis paramyosin (CsPmy) from the cyst wall proteins of metacercariae by proteomic approaches and characterized the expressed recombinant pET-26b-CsPmy protein (101 kDa). Bioinformatics analysis indicated that full-length sequences of paramyosin are conserved in helminthes and numerous B-cell/T-cell epitopes were predicted in amino acid sequence of CsPmy. Western blot analysis showed that CsPmy was expressed at four life stages of C. sinensis, both cyst wall proteins and soluble tegumental components could be probed by anti-CsPmy serum. Moreover, immunolocalization results revealed that CsPmy was specifically localized at cyst wall and excretory bladder of metacercaria, as well as the tegument, oral sucker and vitellarium of adult worm. Both immunoblot and immunolocalization results demonstrated that CsPmy was highly expressed at the stage of adult worm, metacercariae and cercaria, which could be supported by real-time PCR analysis. Both recombinant protein and nucleic acid of CsPmy showed strong immunogenicity in rats and induced combined Th1/Th2 immune responses, which were reflected by continuous high level of antibody titers and increased level of IgG1/IgG2a subtypes in serum. In vaccine trials, comparing with control groups, both CsPmy protein and DNA vaccine exhibited protective effect with significant worm reduction rate of 54.3% (psinensis regarding its high immunogenicity and surface localization. PMID:22470461

Morphological and molecular characterization of the metacercaria of Paragonimus caliensis, as a separate species from P. mexicanus in Costa Rica.

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Hernández-Chea, Roderico; Jiménez-Rocha, Ana Eugenia; Castro, Ruth; Blair, David; Dolz, Gaby

2017-04-01

The trematode Paragonimus mexicanus is the etiological agent of paragonimiasis, a food-borne zoonotic disease in Latin America. This species, as well as Paragonimus caliensis, have been reported from Costa Rica, but it is not known if the two are synonymous. Two types of Paragonimus metacercariae from freshwater pseudothelphusid crabs from several localities in Costa Rica were recognized by light microscopy. Morphologically, these corresponded to descriptions of P. mexicanus and P. caliensis. Metacercariae of the former species lacked a membrane or cyst and their bodies were yellow in color. Those of P. caliensis were contained in a transparent thin cyst and were pink in color. Morphotypes of metacercariae were determined using scanning electron microscopy (SEM). Based on the number and distribution of papillae in the ventral sucker, three morphotypes were found for P. mexicanus and two for P. caliensis. Analysis of DNA sequences (nuclear ribosomal 28S and ITS2 genes, and partial mitochondrial cox1 gene) confirmed the presence of P. mexicanus and provided the first molecular data for P. caliensis. The two species are phylogenetically distinct from each other and distant from the Asian species. The confirmation of P. caliensis as a separate species from P. mexicanus raises several questions about the ecology, biological diversity, and epidemiology of the genus Paragonimus in Costa Rica. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Descriptions of diplostomid metacercariae (Digenea: Diplostomidae from freshwater fishes in the Tshwane area

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Esmey B.E. Moema

2013-09-01

Full Text Available The metacercarial (larval stages of diplostomid digeneans are known to inhabit freshwater fish, causing tissue damage in the process. Due to their widespread diversity, little is known about their life cycle. The classification of these parasitic stages to the species level using only the morphology is very challenging due to the lack of genitalia; they are regarded to be the most important structures in the identification of these organisms. In this study, additional morphological information through light and scanning electron microscopy is given for two different diplostomids found in the cranial cavity of Clarias gariepinus and the vitreous chambers of Tilapia sparrmanii and Pseudocrenilabrus philander. The diplostomid metacercaria inhabiting the cranial cavity of Clarias gariepinus was morphologically identified as Diplostomulum (Tylodelphys mashonenseand an unknown metacercaria of the genus Diplostomumwas found in the vitreous chambers of Pseudocrenilabrus philander and Tilapia sparrmanii. Both parasitic species’ 28S recombinant deoxyribonucleic acid genomic regions were successfully amplified using Dig 125/1500R primer pairs. The assay yielded a product of approximately 1300 base pairs as seen on the gel images. There were 14 nucleotide differences over the entire analysed sequences resulting in a 1.1% (14/1273 nucleotide difference. In line with the morphological characteristics of these parasites, there seemed to be a slight difference in their genetic makeup. The application of molecular techniques on digenetic trematodes seems very promising and may yield great potential in future descriptions of morphologically similar parasitic species.

First report of metacercariae of Cyathocotyle prussica parasitising a fish host in the Czech Republic, Central Europe

Czech Academy of Sciences Publication Activity Database

Kvach, Yuriy; Ondračková, Markéta; Jurajda, Pavel

2016-01-01

Roč. 53, č. 3 (2016), s. 257-261 ISSN 0440-6605 R&D Projects: GA ČR(CZ) GBP505/12/G112; GA ČR(CZ) GAP505/12/2569 Institutional support: RVO:68081766 Keywords : Cyathocotyle prussica * Proterorhinus semilunaris * metacercariae * morphology * first finding Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.472, year: 2016

Infección experimental del hámster (Mesocricetus auratus con metacercarias de Paragonimus mexicanus (peruvianus

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Alina Huiza F

2001-07-01

Full Text Available Un hámster hembra adulta infectada oralmente con 11 metacercarias de Paragonimus mexicanus (peruvianus murió a los 10 días de ser infectado. A la autopsia del animal, se observaron y recuperaron 5 ejemplares juveniles de Paragonimus mexicanus (peruvianus aún móviles: 2 en tejido pulmonar, 2 en cavidad toráxica y 1 en tejido graso del peritoneo abdominal.

Artyfechinostomum malayanum: Metacercariae Encysted in Pila sp. Snails Purchased from Phnom Penh, Cambodia.

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Sohn, Woon-Mok; Yong, Tai-Soon; Eom, Keeseon S; Sinuon, Muth; Jeoung, Hoo-Gn; Chai, Jong-Yil

2017-06-01

The metacercariae of Artyfechinostomum malayanum (Leiper, 1911) Mendheim, 1943 were discovered in Pila sp. snails purchased from a market in Phnom Penh, Cambodia. They were isolated from the snails using the artificial digestion technique and were orally fed to 2 hamsters, 1 rat, and 2 mice to obtain the adult flukes. The metacercariae were round, 145-165 μm in diameter, having a cyst wall of 6-10 μm in thickness, a head collar and collar spines, and characteristic features of excretory granules. Adult flukes were recovered in the small intestines of the animals at days 14 and 32 post infection and were morphologically observed using a light microscope and a scanning electron microscope. They were plump or elongated, ventrally curved, 6.0-8.1 × 1.6-2.0 mm in size, and characterized by the head collar bearing 43 collar spines, including 5 end group ones on each side, a long cirrus sac extending beyond the posterior margin of the ventral sucker, a submedian ovary, and 2 deeply lobed testes. Eggs in uteri were operculate, ovoid to ellipsoid, and 120-135 × 68-75 μm in size. In scanning electron microscopy, the head collar was prominent with collar spines looking like horns. Scale-like tegumental spines were densely distributed on the ventral surface between the head collar and ventral sucker. Sensory papillae were distributed mainly on the tegument around suckers. By this study, it has been first confirmed that the life cycle of A. malayanum exists in Cambodia.

Identification and characterization of paramyosin from cyst wall of metacercariae implicated protective efficacy against Clonorchis sinensis infection.

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Xiaoyun Wang

Full Text Available Human clonorchiasis has been increasingly prevalent in recent years and results in a threat to the public health in epidemic regions, motivating current strategies of vaccines to combat Clonorchis sinensis (C. sinensis. In this study, we identified C. sinensis paramyosin (CsPmy from the cyst wall proteins of metacercariae by proteomic approaches and characterized the expressed recombinant pET-26b-CsPmy protein (101 kDa. Bioinformatics analysis indicated that full-length sequences of paramyosin are conserved in helminthes and numerous B-cell/T-cell epitopes were predicted in amino acid sequence of CsPmy. Western blot analysis showed that CsPmy was expressed at four life stages of C. sinensis, both cyst wall proteins and soluble tegumental components could be probed by anti-CsPmy serum. Moreover, immunolocalization results revealed that CsPmy was specifically localized at cyst wall and excretory bladder of metacercaria, as well as the tegument, oral sucker and vitellarium of adult worm. Both immunoblot and immunolocalization results demonstrated that CsPmy was highly expressed at the stage of adult worm, metacercariae and cercaria, which could be supported by real-time PCR analysis. Both recombinant protein and nucleic acid of CsPmy showed strong immunogenicity in rats and induced combined Th1/Th2 immune responses, which were reflected by continuous high level of antibody titers and increased level of IgG1/IgG2a subtypes in serum. In vaccine trials, comparing with control groups, both CsPmy protein and DNA vaccine exhibited protective effect with significant worm reduction rate of 54.3% (p<0.05 and 36.1% (p<0.05, respectively. In consistence with immune responses in sera, elevated level of cytokines IFN-γ and IL-4 in splenocytes suggested that CsPmy could induce combined cellular immunity and humoral immunity in host. Taken together, CsPmy could be a promising vaccine candidate in the prevention of C. sinensis regarding its high immunogenicity

Effect of infection with Metacercariae of Himasthla elongata (Trematoda: Echinostomatidae) on cardiac activity and growth rate in blue mussels (Mytilus edulis) in situ

Science.gov (United States)

Bakhmet, Igor; Nikolaev, Kirill; Levakin, Ivan

2017-05-01

Trematode parasites can affect their molluscan hosts, which serve as the first intermediate hosts in their life cycles, in manifold ways, but little is known about trematode-induced effects on their second intermediate hosts. Experimental infection of blue mussels Mytilus edulis serving as second intermediate hosts for larval stages (metacercariae) of the trematodes Himasthla elongata was studied in field experiments during one year. The heart rates and growth rates of noninfected mussels were significantly higher than those of infected mussels. During the summer, the heart rates of noninfected mussels showed rhythmic oscillations, whereas the parasitized animals displayed no any rhythmicity. There was a significant difference between the infected and uninfected mussels in relation to heart rates and temperature. The results indicate that mussels infected with H. elongata metacercariae may be at an energetic disadvantage relative to noninfected mussels. Furthermore, trematode infection may disrupt neuronal control of cardiac function.

A rabbit eye model for in vivo transformation of progenetic metacercariae of Clinostomum complanatum into ovigerous adult worms.

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Rizvi, A; Zaidi, Z A; Alam, M M; Zafar, A; Shareef, P A A; Saifullah, M K; Saleemuddin, M; Abidi, S M A

2014-03-01

Clinostomum complanatum is a digenetic trematode that causes yellow grub disease in some fish species and also shows zoonotic potential by sporadically infecting humans. In this study, progenetic metacercariae of C. complanatum were obtained from the fish Trichogaster fasciatus, and were aseptically placed in conjunctival incisions made in the superior and inferior fornices of the eye of rabbits, which served as the experimental hosts. Worms were harvested without necropsy of the host on days 4 and 8 post infection, to observe in vivo transformation of the progenetic metacercariae into ovigerous adult worms. The worms appeared to cause minimal damage to the host although they were tenaciously attached. In vivo maturation was evident by the development of the vitellaria, enlargement of gonads, the presence of a large number of shelled eggs in a distended uterus and ramifications of the intestinal caeca. Obtaining mature ovigerous worms without sacrificing the host clearly gives the rabbit eye model an advantage over those described previously. Due to the relative advantage of the short time required for maturation and the prolific egg production by C. complanatum, it is suggested that this host-parasite system could be used as an excellent model for classroom teaching of trematode biology and to investigate the cues involved in in vivo transformation and host-parasite interactions.

Distribución parcial de Paragonimus (Digenea: Troglotrematidae) en Antioquia, por presencia de metacercarias en cangrejos dulciacuícolas

OpenAIRE

Luz Elena Velásquez; Mónica Uruburu; Mabel Granada

2008-01-01

Introducción. La paragonimosis, o distomatosis pulmonar, es una enfermedad con sintomatología similar a la observada en la tuberculosis. Es causada por parásitos del género Paragonimus (Digenea: Troglotrematidae). Las personas se infectan al consumir cangrejos crudos o mal cocidos, con metacercarias del parásito. El primer foco de paragonimosis humana en Colombia se registró durante 1995 en Urrao, Antioquia, donde se hallaron dos especies de cangrejos que hospedaban el parásito. En el 2005 se...

The Irradiation Effect Against To The Infectivity Of Metacercaria Of Fasciola Gigantica On Goat

International Nuclear Information System (INIS)

Arifin, M.; Boky, J.T.; Tarmizi

2000-01-01

An experiment was carried out to study the effect of irradiation against to the infectivity of F.gigantica by using observation the exchange of goat bodies condition. Four groups of experiment animals were inoculated by irradiated meta cercaria of F. gigantica at a doses of 0, 45, 55 and 65Gy, and the other one is the negative control. Each experiment animal received 350 lives meta cercaria. The infectivity of irradiated meta cercaria in goat has been followed in the development of body weight, blood value described as the number of red blood cells (RBC), level of haemoglobin (Hb), percentages of packed cell volume (PCV), eosinofil and the last is pathology anatomic inspection. The results obtained showed that 45 Gy is the optimal dose of irradiation for decreasing infectivity of metacercariae of F.gigantica which have the ability to stimulate the good immune response in the goat

The effectiveness of different intervention strategies for the prevention of zoonotic metacercariae infection in cultured fish

DEFF Research Database (Denmark)

Clausen, Jesper Hedegaard; Madsen, Henry; Murrell, Kenneth Darwin

2013-01-01

interventions was reduced 91.7% compared to before interventions. The intensity of FZT was also significantly lower in the pond management group, compared to the drug treatment group and the control group after interventions. The results demonstrate that improving farm and pond management practices can......Fish-borne zoonotic trematodes (FZT) are a major public health problem. It is estimated that in Vietnam 26,366 "disability-adjusted life years" (DALYs) are lost due to FZT. Fish from aquaculture are a main source of protein and of great economic importance in both rural and urban areas...... with two intervention groups; a drug treatment of human and animal groups and a farm management group (control of snail vectors and fecal pollution of pond). A third group (non-intervention) served as control. Fish were examined for FZT metacercariae prevalence and intensity before and after interventions...

Immunization of zebu calves against Fasciola gigantica, using irradiated metacercariae

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Younis, S.A.; Yagi, A.I.; Haroun, E.M.; Gameel, A.A.; Taylor, M.G.

1986-06-01

The pathogenesis of unirradiated, 3 krad-irradiated and 20 krad-irradiated metacercarial infections was compared in zebu calves studied over 10 weeks. Calves exposed to 1000 unirradiated metacercariae (MC) became hypoalbuminaemic, and showed elevated serum concentrations of liver enzymes, whereas neither of the other groups was significantly affected. At slaughter, a mean of 332 flukes was recovered from the 0 krad group, while only 23% and 12% of this number were recovered from the 3 krad and 20 krad groups, respectively. All the worms recovered from the 20 krad group were stunted, but a mean of 13% of the flukes recovered from the 3 krad group were large. Liver lesions typical of acute fascioliasis were present in the 0 krad group, but lesions in the other groups, were far less severe. A single vaccination of calves with 10003 krad-irradiated mc induced partial resistance to a challenge with 1000 normal mc eight weeks later, but the reduction in worm recovery was not statistically significant. There was less evidence of protection when two vaccinating doses of 3 krad mc were given within four weeks, with challenge at week 8, and a single vaccination was ineffective against a challenge four weeks later. However, when the irradiation dose was increased to 20 krad, a high level of resistance was induced by a single vaccination, given eight weeks before challenge, and liver pathology was strikingly reduced in the vaccinated animals.

Immunization of zebu calves against Fasciola gigantica, using irradiated metacercariae

International Nuclear Information System (INIS)

Younis, S.A.; Yagi, A.I.; Haroun, E.M.; Gameel, A.A.; Taylor, M.G.

1986-01-01

The pathogenesis of unirradiated, 3 krad-irradiated and 20 krad-irradiated metacercarial infections was compared in zebu calves studied over 10 weeks. Calves exposed to 1000 unirradiated metacercariae (mc) became hypoalbuminaemic, and showed elevated serum concentrations of liver enzymes, whereas neither of the other groups was significantly affected. At slaughter, a mean of 332 flukes was recovered from the 0 krad group, while only 23% and 12% of this number were recovered from the 3 krad and 20 krad groups, respectively. All the worms recovered from the 20 krad group were stunted, but a mean of 13% of the flukes recovered from the 3 krad group were large. Liver lesions typical of acute fascioliasis were present in the 0 krad group, but lesions in the other groups, were far less severe. A single vaccination of calves with 10003 krad-irradiated mc induced partial resistance to a challenge with 1000 normal mc eight weeks later, but the reduction in worm recovery was not statistically significant. There was less evidence of protection when two vaccinating doses of 3 krad mc were given within four weeks, with challenge at week 8, and a single vaccination was ineffective against a challenge four weeks later. However, when the irradiation dose was increased to 20 krad, a high level of resistance was induced by a single vaccination, given eight weeks before challenge, and liver pathology was strikingly reduced in the vaccinated animals. (author)

Resistance to reinfection in rats induced by irradiated metacercariae of Clonorchis sinensis

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Fu Shi Quan

2005-08-01

Full Text Available A study was made to observe the association between the resistance to reinfection induced by irradiated metacercariae (MC of Clonorchis sinensis and antigen specific Th1- and Th2-type cytokine productions in rats. Rats were infected with 20 MC of C. sinensis, previously exposed to a single dose of gamma irradiation, which varied from 0 to 100 Gy. All of them, single dose of 12 Gy showed higher IgG antibody titer with lowest worm recovery. Thus, 50 MC were used to challenge infection in rats previously infected with 20 MC irradiated at 12 Gy and the highest resistance to challenge infection was observed. The results of lymphocyte proliferation with specific antigen, ES Ag were shown no difference of proliferative responses as compared with primary and challenge infection at 12 Gy irradiation dose. In the case of cytokines production were observed that interferon (IFN-gamma and interlukin (IL-2 were significantly enhanced, while IL-4 and IL-10 was almost unchanged to make comparison between primary and secondary infection at 12 Gy irradiation dose. In conclusion, the single dose of 12 Gy could be adopted for induction of the highest resistance to challenge infection. Up-regulation of Th1 type cytokines, IFN-gamma and IL-2 may be affected to develop vaccine by irradiated MC.

Inactivation of Paragonimus westermani metacercariae in soy sauce-marinated and frozen freshwater crabs.

Science.gov (United States)

Kim, Tae Im; Oh, Se-Ra; Dai, Fuhong; Yang, Hyun-Jong; Ha, Sang-Do; Hong, Sung-Jong

2017-03-01

Soy sauce-marinated freshwater crabs (Eriocheir japonicus) are a source of human paragonimiasis. The viability of Paragonimus westermani metacercariae (PwMc) in marinated crabs was investigated in an experimental setting. The PwMc collected from freshwater crayfish were inoculated into freshwater crabs, which were then frozen or marinated in soy sauce. All PwMc in the freshwater crabs were inactivated after freezing for 48 h at -20 °C and after freezing for 12 h at -40 °C. After marinating for 32 days, the survival rate of PwMc in 5% NaCl soy sauce was 50%, in 7.5% NaCl soy sauce it was 33.3%, and in 10.0% NaCl soy sauce it was 31.3%. When marinated for 64 days, all PwMc were inactivated in all experimental groups. These results revealed that freezing and soy sauce marination were detrimental to the survival of PwMc in freshwater crabs. Specifically, freezing crabs for more than 48 h or soaking them in soy sauce containing at least 5.0% NaCl for 64 days can inactivate PwMc. These results can inform the production of the traditional Korean soy sauce-marinated freshwater crabs known as gejang.

INFECCIÓN DE HYPOLOBOCERA CHILENSIS EIGENMANI POR METACERCARIAS DE PARAGONIMUS MEXICANUS (= PERUVIANUS EN EL DISTRITO DE CONDEBAMBA (CAJAMARCA, PERÚ

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Alina Huiza F.

2014-06-01

Full Text Available Cangrejos de rio Hypolobocera chilensis eigenmanni fueron colectados de acequias durante el año 1997 en estación seca (mayo a diciembre en Chaquicocha, Área que pertenece al distrito de Condebamba (departamento de Cajamarca en la parte norte del Perú. Ciento treinta y un cangrejos colectados fueron transportados al Laboratorio de Parasitología y examinados por disección, 27 de 131 (20,6% estaban infectados por metacercarias de Paragonimus mexicanus(=peruvianus. La intensidad de la infección fue de 1 a 5 en la mayoría de los; casos (81,5% con un promedio de 4,85 por cangrejo. Estos datos son diferentes; a los de estudios anteriores; en la misma Área donde fueron más; altos, lo que indica una tendencia al decrecimiento del número de cangrejos infectados.

Molecular analysis of echinostome metacercariae from their second intermediate host found in a localised geographic region reveals genetic heterogeneity and possible cryptic speciation.

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Waraporn Noikong

2014-04-01

Full Text Available Echinostome metacercariae are the infective stage for humans and animals. The identification of echinostomes has been based until recently on morphology but molecular techniques using sequences of ribosomal RNA and mitochondrial DNA have indicated major clades within the group. In this study we have used the ITS2 region of ribosomal RNA and the ND1 region of mitochondrial DNA to identify metacercariae from snails collected from eight well-separated sites from an area of 4000 km2 in Lamphun Province, Thailand. The derived sequences have been compared to those collected from elsewhere and have been deposited in the nucleotide databases. There were two aims of this study; firstly, to determine the species of echinostome present in an endemic area, and secondly, to assess the intra-specific genetic diversity, as this may be informative with regard to the potential for the development of anthelmintic resistance and with regard to the spread of infection by the definitive hosts. Our results indicate that the most prevalent species are most closely related to E. revolutum, E. trivolvis, E. robustum, E. malayanum and Euparyphium albuferensis. Some sites harbour several species and within a site there could be considerable intra-species genetic diversity. There is no significant geographical structuring within this area. Although the molecular techniques used in this study allowed the assignment of the samples to clades within defined species, however, within these groupings there were significant differences indicating that cryptic speciation may have occurred. The degree of genetic diversity present would suggest the use of targeted regimes designed to minimise the selection of anthelmintic resistance. The apparent lack of geographic structuring is consistent with the transmission of the parasites by the avian hosts.

Infeccion natural de Ptychophallus tristani (Crustacea: decapoda con metacercarias de Paragonimus mexicanus (Trematoda en Tabarcia de Mora, Costa Rica

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Eduardo Monge

1985-02-01

Full Text Available Se determinó el grado de infección de los crustáceos Ptychophallus tristani presentes en una área endémica de Paragonimus mexicanus, encontrándose que de un total de 182 cangrejos, 161 resultaron positivos (88,5% por metacercarias. Al comparar la infección entre machos y hembras no se encontró diferencias estadísticamente significativas y en relación a su tamaño se observó una relación directa entre tamaño y porcentaje y grado de infección. Se estableció que no existen diferencias significativas en relación a la época del año en que los cangrejos fueron capturados lo que epidemiológicamente es importante pues indica que las fuentes de infección permanecen potencialmente activas durante todo el año.

Metacercárias de Neascus sp. em Geophagus brasiliensis (Perciformes: Cichlidae do rio do Peixe, Juiz de Fora, Brasil - DOI: 10.4025/actascibiolsci.v30i3.493 Neascus-type metacercariae in Geophagus brasiliensis (Perciformes: Cichlidae of Peixe river, Juiz de Fora, Brazil - DOI: 10.4025/actascibiolsci.v30i3.493

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José Luis Luque

2008-10-01

Full Text Available A metacercária de Neascus sp., causadora da doença dos pontos pretos, é descrita em espécimes de Geophagus brasiliensis coletados no Brasil. Entre março e maio de 2006, foram capturados 33 espécimes de G. brasiliensis, coletados com o auxílio de vara e anzol no Rio do Peixe, Monte Verde, município de Juiz de Fora, Estado de Minas Gerias. Dos 33 espécimes examinados, 30 (90,9% apresentaram-se parasitados por metacercárias causadoras dos pontos pretos, com intensidade média de infecção de 15,6 ± 16,1 (1 – 75. Foi observada correlação significativa entre o comprimento total de G. brasiliensis e a abundância parasitária (r = 0,459; P Neascus-type metacercariae, which cause the black spot disease, are described in specimens collected from 33 samples of Geophagus brasiliensis from the Peixe river, in Monte Verde, Juiz de Fora, state of Minas Gerais, Brazil, during March and May 2006. The main morphological features of these metacercariae are: genital primordia composed of three cell masses located in the hind body, and a median dorsal vessel of the reserve bladder around the acetabulum. Thirty fish specimens (90.9% were parasitized by metacercariae, with mean infection intensity of 15.6 ± 16.1 (1-75. Also, a positive correlation was detected between G. brasiliensis total length and the parasite abundance (r = 0.459; p < 0.01.

COMPARAÇÃO ENTRE OS MÉTODOS DE EXTRAÇÃO DE METACERCÁRIAS DE ASCOCOTYLE SP (TREMATODA: DIGENEA DOS TECIDOS DE MUGIL LIZA VALENCIENNES, 1836 (TELEOSTEI: MUGILIDAE

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Renato Ribeiro Nogueira Ferraz

2014-09-01

Full Text Available The demand and consumption of fish and their derivatives has increased considerably in recent years. However, fish are ideal hosts of numerous parasites, highlighting the need to develop new research methodologies for its detection. The aim of present study was to compare the efficacy of Ascocotyle metacercariae (Trematoda: Digenea extraction from visceral tissues of Mugil liza Valenciennes, 1836 (Teleostei: Mugilidae by two methods: homogenization by blender or mixer. Twentysix samples of M. liza were collected, being 16 liver samples and 10 samples of muscle tissue. Approximately 5g of each sample were processed by blender and mixer techniques homogenization for metacercariae extraction. In liver samples, up to 46 metacercariae were found in samples homogenized in blender. The lowest amount found was 2 metacercariae for blender and mixer techniques. In samples of muscle tissue, 4 metacercariae were observed in the mixer extraction. The lowest amount was found to be 2 parasites to blender and mixer. The mean metacercariae found and extracted from muscle tissue were 0.2 (+0.357 and 1.2 (+0.963 for blender and mixer, respectively. The averages of metacercariae found and extracted from fish liver, in blender and mixer, were 24 (+15.145 and 18 (+8.246, respectively. The homogenization techniques for blender and mixer were effective for the extraction of metacercariae of mullet fish tissues, suggesting that they may be directly applicable in the field of study, especially due to the ease of testing.

Prevalence and Intensity of Clonorchis sinensis Metacercariae in Freshwater Fish from Wicheon Stream in Gunwi-gun, Gyeongsangbuk-do, Korea

Science.gov (United States)

Sohn, Woon-Mok; Na, Byoung-Kuk; Cho, Shin-Hyeong; Ju, Jung-Won; Son, Dong-Chul

2018-01-01

The infection status of Clonorchis sinensis metacercariae (CsMc) was examined in freshwater fish from a highly prevalent site, Wicheon (a branch of Nakdong-gang), which is located in Gunwi-gun, Gyeongsangbuk-do, the Republic of Korea. Total 1,162 fish in 32 species were examined by the artificial digestion method through 6 years. CsMc were detected in 720 (67.5%) out of 1,067 fish (26 spp.) and their density was 610 per fish infected. In the susceptible gobioninid fish group, i.e., Pungtungia herzi, Squalidus gracilis majimae, Squalidus japonicus coreanus, Sarcocheilichthys variegatus wakiyae and Pseudorasbora parva, all of 323 fish were infected with an average of 1,310 CsMc. Total 23 (95.8%) gobioninid fish, i.e., Pseudogobio esocinus, Abbottina springeri, Hemibarbus longirostris, Microphysogobio koreensis, and Microphysogobio jeoni, were infected with 127 CsMc in average. In the acheilognathinid fish (bitterlings) group, the prevalence was 77.0%, and the density was 50 CsMc per fish infected. In the rasborinid fish (chubs) group, i.e., Zacco platypus, Zacco temminckii, Zacco koreanus, and Opsariichthys uncirostris amurensis, 147 (36.5%) out of 403 fish examined were infected with 15 CsMc in average. The susceptibility indices of CsMc were 412 in the overall positive fish group, 1,310 in the gobioninid group-1, 122 in the gobioninid group-2, 38.5 in the acheilognathinid group, and 5.5 in the rasborinid fish group. Conclusively, it was confirmed that CsMc are highly prevalent in fish from Wicheon, and their infection tendency varied according to the subfamily groups in Cyprinidae fish hosts. PMID:29529849

Comparative study of the sensibility of Metacercaria cysts of Phagicola Faust, 1920 (Trematoda: Heterophyidae) to ionizing radiation and to freezing in raw mullets Mugil Linnaeus, 1758 (Pisces: Mugilidae); Estudo comparativo da sensibilidade de cistos de Metacercarias de Phagicola Faust, 1920 (Trematoda: Heterophyidae) a radiacao ionizante e ao congelamento em peixes crus preparados a partir de tainha Mugil Linnaeus, 1758 (Pisces: Mugilidae)

Energy Technology Data Exchange (ETDEWEB)

Moraes, Ivany Rodrigues de

2005-07-01

Foodborne illnesses have been a growing concern to the governments, international organizations and consumers, mainly regarding the damages they cause to human health. This issue is the result of the substitution of imported food habits, which enable the consumption of raw fish or wrongly prepared fish. Among the emergent parasitic zoonosis, the Fagicolose stands out. It is transmitted by Phagicola longa, a trematoda of great importance in public health and responsible for high percentages of Mugilidae infections, that is able to produce humans' parasitism. In this experiment, the studies were focused to the radio sensibility of metacercaria of Phagicola longa exposed to a treatment with ionizing radiation (gamma rays and electron beams) and the sensibility to freezing, in artificially infected fish. Some data were acquired regarding the occurrence of Phagicola longa in mullets bought in the local market, the freezer temperature of 10 domestic refrigerators, and the origin and preservation of the mullets in some Japanese-type restaurants in Sao Paulo. The results indicated more effectiveness in the treatment with the electron accelerator, as well as with the freezing at the temperature of-16 deg C. (author)

Comparative study of the sensibility of Metacercaria cysts of Phagicola Faust, 1920 (Trematoda: Heterophyidae) to ionizing radiation and to freezing in raw mullets Mugil Linnaeus, 1758 (Pisces: Mugilidae)

International Nuclear Information System (INIS)

Moraes, Ivany Rodrigues de

2005-01-01

Foodborne illnesses have been a growing concern to the governments, international organizations and consumers, mainly regarding the damages they cause to human health. This issue is the result of the substitution of imported food habits, which enable the consumption of raw fish or wrongly prepared fish. Among the emergent parasitic zoonosis, the Fagicolose stands out. It is transmitted by Phagicola longa, a trematoda of great importance in public health and responsible for high percentages of Mugilidae infections, that is able to produce humans' parasitism. In this experiment, the studies were focused to the radio sensibility of metacercaria of Phagicola longa exposed to a treatment with ionizing radiation (gamma rays and electron beams) and the sensibility to freezing, in artificially infected fish. Some data were acquired regarding the occurrence of Phagicola longa in mullets bought in the local market, the freezer temperature of 10 domestic refrigerators, and the origin and preservation of the mullets in some Japanese-type restaurants in Sao Paulo. The results indicated more effectiveness in the treatment with the electron accelerator, as well as with the freezing at the temperature of-16 deg C. (author)

Antibody fluctuations of infected cattle with Fasciola gigantica and the effect of triclabendazole treatment

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S Widjajanti

2001-12-01

Full Text Available Observation on the antibody fluctuations of infected cattle with metacercariae of Fasciola gigantica and the effect of triclabendazole treatment were made by means of ELISA technique. Seven cattle were infected with 700 metacercariae and one cattle remained uninfected, as negative control animal. Treatment with triclabendazole was given to 6 cattle, when the mean antibody levels of infected cattle reached the peak, and the other one remained untreated, as positive control animal. One week after treatment the mean antibody levels started to drop and then decreased gradually. After eight weeks of treatment, the mean antibody levels of the treated cattle reached the lowest level or the same value as before infection, thereafter, 6 cattle were reinfected with different dosages of metacercariae of F. gigantica. Two cattle were infected with 400 metacercariae, the other two were infected with 600 metacercariae and the rest of them were infected with 800 metacercariae. The results showed that the immunological responses of re-infected cattle are quicker (5 weeks after infection and the peak of the antibody levels are higher (ELISA OD = 1.7 than after the first infection (11 weeks after infection and ELISA OD = 1.2. However, after re-infection, there were no significant different on the antibody fluctuations and antibody levels among the infected group, although those cattle received different dosages.

Effects of gamma irradiation on the survival and development of Gymnophalloides seoi in C3H mice. Final report for the period 1 November 1994 - 31 October 1995

International Nuclear Information System (INIS)

Chai Yong-Vil

1996-01-01

Gymnophalloides seoi is a peculiar human intestinal trematode in Korea transmitted by oysters. This study was carried out to observe the effects of radiation on the infectivity of G. seoi metacercariae to C3H mice and to assess the applicability of radiation for use in the control of gymnophalloidiasis. Oysters were collected from the endemic area. Non-irradiated control, metacercaria-irradiation, and oyster-irradiation groups were prepared. One hundred metacercariae were infected orally to each mouse, and worm recovery rates of three groups were compared at the seventh day post-infection. In the metacercaria-irradiation group, the worm recovery rate was significantly reduced at radiation doses higher than 200 Gy, and the number of intrauterine eggs was significantly reduced at doses over 50 Gy. In the oyster-irradiation group, 50 Gy significantly reduced the worm recovery rate and number of uterine eggs. In the two groups, no worm was recovered at 1,000 Gy. In conclusion, G. seoi metacercariae showed some resistance to radiation at lower doses than 200 Gy, but irradiation of oyster with 200-1,000 Gy could be applied as a control measure for gymnophalloidiasis. (author). 10 refs, 3 figs, 2 tabs

Usefulness of high-resolution sonography in early diagnosis of rabbit clonorchiasis

International Nuclear Information System (INIS)

Lim, Jae Hoon; Choi, Don Gil; Chung, Il Gyu; Phyun, Lae Hyun; Pyeun, Yong Seon; Hong, Sung Tae; Lee, Me Jeong

1999-01-01

To determine the role of high-resolution sonography in the early diagnosis of experimentally induced clonorchiasis in rabbits. We performed sonographic examination weekly in 22 lightly-infected rabbits (10 rabbits infected with 10 metacercariae, 6 rabbits infected with 20 metacercariae, and 6 rabbits infected with 40 metacercariae), and 10 heavily-infected rabbits (500 metacercariae). The sonographic criterion of diagnosis with dilatation of the intrahepatic ducts. We sacrificed lightly-infected rabbits and counted numbers of adult worms of clonorchis sinensis 9 weeks after infection. Sonographic abnormalities were found 3 weeks after infection in 2 lightly-infected rabbits and 5 heavily-infected rabbits. On sonography at 9 weeks after infection, we observed dilatation of the intrahepatic ducts in 11 (65%) of 17 lightly-infected rabbits and all of 10 heavily-infected rabbits. High-resolution sonography is very useful in early diagnosis of rabbits clonorchiasis.

Usefulness of high-resolution sonography in early diagnosis of rabbit clonorchiasis

Energy Technology Data Exchange (ETDEWEB)

Lim, Jae Hoon; Choi, Don Gil; Chung, Il Gyu; Phyun, Lae Hyun; Pyeun, Yong Seon [Sungkyunkwan University College of Medicine, Seoul (Korea, Republic of); Hong, Sung Tae; Lee, Me Jeong [Seoul National University College of Medicine, Seoul (Korea, Republic of)

1999-09-15

To determine the role of high-resolution sonography in the early diagnosis of experimentally induced clonorchiasis in rabbits. We performed sonographic examination weekly in 22 lightly-infected rabbits (10 rabbits infected with 10 metacercariae, 6 rabbits infected with 20 metacercariae, and 6 rabbits infected with 40 metacercariae), and 10 heavily-infected rabbits (500 metacercariae). The sonographic criterion of diagnosis with dilatation of the intrahepatic ducts. We sacrificed lightly-infected rabbits and counted numbers of adult worms of clonorchis sinensis 9 weeks after infection. Sonographic abnormalities were found 3 weeks after infection in 2 lightly-infected rabbits and 5 heavily-infected rabbits. On sonography at 9 weeks after infection, we observed dilatation of the intrahepatic ducts in 11 (65%) of 17 lightly-infected rabbits and all of 10 heavily-infected rabbits. High-resolution sonography is very useful in early diagnosis of rabbits clonorchiasis.

Presence of three distinct genotypes within the Paragonimus westermani complex in northeastern India.

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Rekha Devi, K; Narain, Kanwar; Mahanta, Jagadish; Nirmolia, Tulika; Blair, David; Saikia, Sidhartha P; Agatsuma, Takeshi

2013-01-01

The name Paragonimus westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus westermani complex in NE India. Two of these were previously unknown.

Sphincterodiplostomum musculosum (Digenea, Diplostomidae) infecting Steindachnerina insculpta (Characiformes, Curimatidae) in the Chavantes Reservoir, Southeastern Brazil.

Science.gov (United States)

Zago, Aline Cristina; Franceschini, Lidiane; Ramos, Igor Paiva; Zica, Erica de Oliveira Penha; Wunderlich, Alison Carlos; Carvalho, Edmir Daniel; Silva, Reinaldo José da

2013-01-01

This study aimed to report the infection by Sphincterodiplostomum musculosum metacercariae in Steindachnerina insculpta from the Chavantes Reservoir, medium Paranapanema River, municipality of Ipaussu, State of São Paulo, Brazil. Thirty specimens of S. insculpta were collected and 29 were infected with S. musculosum metacercariae (prevalence = 96.67%) in the eyes and visceral cavity. The mean intensity of infection and abundance were 96.6 ± 29.41 (7-846) and 93.3 ± 28.6 (0-846), respectively. Positive correlation was observed between parasite abundance in the eyes and standard length (rs = 0.5, p = 0.005), total weight (rs = 0.649, p = 0.0001), and condition factor (rs = 0.439, p = 0.0154). The high parasitism rates by S. musculosum metacercariae in S. insculpta can be an indicative that this fish species is highly susceptible to infection by this diplostomid, and even reflect the presence of a large abundance of the intermediate host. Moreover, S. musculosum metacercariae are reported for the first time in S. insculpta.

Echinostoma revolutum: freshwater snails as the second intermediate hosts in Chiang Mai, Thailand.

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Chantima, Kittichai; Chai, Jong-Yil; Wongsawad, Chalobol

2013-04-01

The occurrence of 37-collar spined echinostome metacercariae in freshwater snails was investigated in 6 districts of Chiang Mai Province, Thailand, from October 2011 to April 2012. A total of 2,914 snails that belong to 12 species were examined, and 7 snail species (Clea helena, Eyriesia eyriesi, Bithynia funiculata, Bithynia siamensis siamensis, Filopaludina doliaris, Filopaludina sumatrensis polygramma, and Filopaludina martensi martensi) were found infected with echinostome metacercariae. The prevalence of metacercariae was the highest in Filopaludina spp. (38.5-58.7%) followed by B. funiculata (44.0%), E. eyriesi (12.5%), B. siamensis siamensis (8.2%), and C. helena (5.1%). Metacercariae were experimentally fed to hamsters and domestic chicks, and adult flukes were recovered from both hosts at days 15 and 20 post-infection. The adult flukes were identified based on morphological features, morphometrics, host-parasite relationships, and geographical distribution. They were compatible to Echinostoma revolutum or Echinostoma jurini, with only minor differences. As the adults were recovered from both hamsters and chicks, our specimens were more compatible to E. revolutum rather than E. jurini (reported only from mammals). This is the first report for metacercariae of E. revolutum in the snail host, C. helena, and also confirmed that Filopaludina spp., E. eryresi, and Bithynia spp. act as the second intermediate hosts of E. revolutum under natural conditions, which are indigenously distributed in Chiang Mai province.

The importance of wild fish in the epidemiology of Clonorchis sinensis in Vietnam.

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Bui, Thanh Ngoc; Pham, Thanh Thi; Nguyen, Nguyen Thi; Nguyen, Ha Van; Murrell, Darwin; Phan, Van Thi

2016-09-01

Preliminary findings of a high prevalence of Clonorchis sinensis in wild-caught fish in a North Vietnam reservoir (Thac Ba reservoir, Yen Bai Province) prompted a longitudinal epidemiological study of fish infections. Monthly collections of fish from September 2014 to August 2015 were processed for recovery of metacercariae; 1219 fish, representing 22 species, were examined. Seven species were infected with C. sinensis metacercariae. Four species, Toxabramis houdemeri, Hemiculter leucisculus, Cultrichthys erythropterus, and Culter recurvirostris, had high prevalence (31.1 to 76.7 %); metacercarial intensities ranged from 3.9 to 65.7 metacercariae/fish. A seasonal variation of C. sinensis prevalence was observed in T. houdemeri. Variation in intensity of infection occurred in C. erythropterus and H. leucisculus. Intensity and prevalence of C. sinensis in the most highly infected species, T. houdemeri, varied by fish size; prevalence was higher in fish weighing more than 3 g, and intensity was higher in fish weighing more than 5 g. The distribution of metacercariae in the body region of T. houdemeri was significantly higher in the caudal fin (14.7 metacercariae/g), compared to the body and head regions (0.7 and 1.4 metacercariae/g, respectively). Further epidemiological investigations on C. sinensis in this reservoir region should include assessing the relative risk of the different fish species for humans based on the latter's food preferences, and the prevalence of C. sinensis in the community. The snail intermediate host(s) in the reservoir should also be identified along with the ecological factors influencing its exposure to C. sinensis eggs and its subsequent transmission of cercariae to fish. Also needed are investigations on the relative importance of wild and domestic reservoir hosts as sources of egg contamination of the reservoir.

Effect of gamma irradiation on different stages of Fasciola hepatica

Energy Technology Data Exchange (ETDEWEB)

Torres, J.L.; Chiriboga, J.

1976-04-01

Fascioliasis or hepatic distomatosis is one of the most widespread liver diseases throughout the world. It is caused by a parasitic worm of the class Trematoda of the phylum Platyhelminthes. This flatworm is commonly known as liver-fluke in the United States and Europe; babosa del higado in Cuba; cucaracha del higado in Puerto Rico; saguaype in Argentina and Chile; pirihuin in Chile; yuta in North Chile, and it is taxonomically classified as Fasciola hepatica. The adult trematode is a common parasite of warm blooded animals especially cattle, sheep, and goats. Alterations in the viability of normal Fasciola hepatica metacercariae as a result of time of storage at 4/sup 0/C were studied by the method of Wikerhauser (1960). A significant decrease in viability was observed only after one hundred days of storage. The in vitro viability of normal metacercariae was compared with that of metacercariae that were exposed to increasing ..gamma.. radiation doses. Two plainly opposite effects observed were: the activation of the excystation process by the lower radiation doses (1.5 and 2.5 Kr), an effect that persisted up to the 14th day post-radiation, and the impairment of the process evidenced on the 14th day in the metacercariae exposed to 3.5 and 5.0 Kr. Correlation between liver damage and the increasing radiation doses to which the metacercariae were exposed was studied up to 34 days after the oral inoculation in rats. (auth)

Effect of gamma irradiation on different stages of Fasciola hepatica

International Nuclear Information System (INIS)

Torres, J.L.; Chiriboga, J.

1976-04-01

Fascioliasis or hepatic distomatosis is one of the most widespread liver diseases throughout the world. It is caused by a parasitic worm of the class Trematoda of the phylum Platyhelminthes. This flatworm is commonly known as liver-fluke in the United States and Europe; babosa del higado in Cuba; cucaracha del higado in Puerto Rico; saguaype in Argentina and Chile; pirihuin in Chile; yuta in North Chile, and it is taxonomically classified as Fasciola hepatica. The adult trematode is a common parasite of warm blooded animals especially cattle, sheep, and goats. Alterations in the viability of normal Fasciola hepatica metacercariae as a result of time of storage at 4 0 C were studied by the method of Wikerhauser (1960). A significant decrease in viability was observed only after one hundred days of storage. The in vitro viability of normal metacercariae was compared with that of metacercariae that were exposed to increasing γ radiation doses. Two plainly opposite effects observed were: the activation of the excystation process by the lower radiation doses (1.5 and 2.5 Kr), an effect that persisted up to the 14th day post-radiation, and the impairment of the process evidenced on the 14th day in the metacercariae exposed to 3.5 and 5.0 Kr. Correlation between liver damage and the increasing radiation doses to which the metacercariae were exposed was studied up to 34 days after the oral inoculation in rats

Larval trematodes (Digenea of the great pond snail, Lymnaea stagnalis (L., (Gastropoda, Pulmonata in Central Europe: a survey of species and key to their identification

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Faltýnková A.

2007-03-01

Full Text Available A survey of cercariae and metacercariae (Trematoda, Digenea from the great pond snail (Lymnaea stagnalis in Central Europe (Austria, Czech Republic, South-East Germany, Poland and Slovak Republic is presented, based on a study of 3,628 snails examined from 1998 to 2005. A total of 953 (26.3% L. stagnalis were infected with 24 trematode species comprising 19 species of cercariae and 11 species of metacercariae (six species occurred both as cercarie and metacercarie of eight families. The dominant cercariae were those of Opisthioglyphe ranae (159 hosts infected, Plagiorchis elegans (141 (both family Plagiorchiidae and Echinoparyphium aconiatum (153 (Echinostomatidae; 14 double infections were found. The most frequent metacercariae were those of Neoglyphe locellus (71 (Omphalometridae, E. aconiatum (66, Echinostoma sp. (59 and Moliniella anceps (48 (Echinostomatidae. In the previous studies carried out in Central Europe, a very similar spectrum of nine trematode families of 22 cercariae determined to species level and 43 types of cercariae reported under generic or provisional names, which can be in many cases conspecific with the previous taxa, were found. A simple key to identification of cercariae and metacercariae, together with their illustrations, is provided.

Effects of fermentation time and low temperature during the production process of Thai pickled fish (pla-som) on the viability and infectivity of Opisthorchis viverrini metacercariae.

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Onsurathum, Sudarat; Pinlaor, Porntip; Haonon, Ornuma; Chaidee, Apisit; Charoensuk, Lakhanawan; Intuyod, Kitti; Boonmars, Thidarut; Laummaunwai, Porntip; Pinlaor, Somchai

2016-02-02

Contamination of a popular fermented fish dish, pla-som, by Opisthorchis viverrini metacercariae (OVMC) is a possible cause of carcinogenic liver fluke infection in Thailand. Affected individuals are at risk of bile duct cancer, which is a major health problem for people in the Greater Mekong Subregion. In order to investigate concerns about food safety, we studied the effects of fermentation time and low temperature on the viability and infectivity of OVMC during the pla-som production process. Pla-som was prepared at room temperature for up to 1 week in duplicate experiments using cyprinid freshwater fish obtained from an O. viverrini-endemic area. OVMC were then isolated and identified under a stereomicroscope. Complete and viable OVMC were found on days 1-4 of fermentation, while their morphology was degenerated thereafter. After OVMC were fed to hamsters, the percentage of the worm recovery after 1 to 2 months of infection was 52%, 44.7%, 11.3% and 1% for days 1, 2, 3 and 4, respectively. In order to measure the effect of low temperature on OVMC, fish were kept in a refrigerator (4 °C) for up to five days and then subsequently fermented for three days. In fish stored in a refrigerator for 1 and 2 days, viable OVMC were clearly observed and were able to infect hamsters, a worm-recovery percentage of 3.3% and 12.7%, respectively. By contrast, in pla-som prepared from fish stored for 3 to 5 days, OVMC were degenerated and could not infect the host. In conclusion, pla-som fermentation for more than four days and refrigerating fish for three days before pla-som processing can prevent O. viverrini infection. This study may increase awareness of fermented-fish dish preparation to prevent liver fluke infection. Copyright © 2015 Elsevier B.V. All rights reserved.

Commercial and game fish from the Dąbie lake (Poland and their infection of Paracoenogonimus ovatus (Digenea

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Angelika Anna Linowska

2015-11-01

Full Text Available Paracoenogonimus ovatus is the parasite that infest wide definitive host range (predator birds and at least two intermediate hosts: molluscs (Viviparus viviparus, V. fasciatus and freshwater fish from eastern and central Europe. The larvae of the parasite (metacercariae go to the muscle and connective tissue, and at high intensity of infection to the internal organs and brain of fish. A total of 128 fishes were caught in autumn 2009 and 2014, by fishing boats operating in Lake Dąbie, which is one of the largest lakes in Poland. The species selected reflect the species structure of commercial and recreational catches. They were roach, bream, silver bream, crucian carp (Cyprinidae, ruffe, perch, zander (Percidae and pike (Esocidae. The fish were filleted, and the skinned muscle was compressed under a trichinoscope, what allowed to see and count metacercariae in the tissues. To facilitate parasite isolation from the cysts, the muscle tissues were subjected to a 0.1 % solution of activated pepsin and 5 % citric acid at a temperature of 20ºC. The free larvae were measured and identified under an Olympus BX 50 microscope coupled with a camera running AxioVs40 V 4.8.2.0 software. Metacercariae are grouped primarily at depths ranging from just beneath the skin to about 50% of fillet thickness. The infection prevalence of P. ovatus metacercariae in the fish analyzed was 78,5%. The density of infection per gram of muscle tissue mass in the fish infected was 8.85 (from 1 to 84 metacercariae. The highest density was noted in roach, bream and white bream (17.09, 11.14 and 9.58, respectively. Metacercariae occurred singly in pike, parasites were not found in the muscles of crucian carp, ruffe, and perch examined. The reason for such differences in infection is the availability and composition of food, as well as the behavior of the fish examined.

Redescription of Austrodiplostomum compactum (Trematoda: Diplostomidae) from its Type Host and Locality in Venezuela, and of Austrodiplostomum mordax from Argentina.

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de Núñez, Margarita Ostrowski

2017-10-01

Austrodiplostomum compactum from Nannopterum brasilianus, and its metacercaria from Geophagus sp. and Oreochromis mossambicus captured (1979) at its type locality, Valencia Lake, Venezuela, by the author, are redescribed. The adult is characterized by its large body size, and an oral sucker smaller than the pharynx. The metacercaria has a similar body size as the adult, and the small genital primordia occupy 4.1-7.3% of body length. Experimental infections in chickens with metacercariae of Diplostomulum mordax from brains of Odonthestes bonariensis, captured (2015) at Dique Paso de las Piedras, near Bahia Blanca City, Buenos Aires Province, Argentina, resulted in 10 adults 5 days postexposure. These adults correspond to Austrodiplostomum mordax as described from N. brasilianus at Lacombe Lagoon, Buenos Aires Province, and differ from A. compactum in their smaller body size, and an oral sucker larger than the pharynx. The metacercaria has a similar body size as the adult and differs mainly in that the larger genital primordia occupy 11.6-13.8% of body length. The status of earlier published Austrodiplostomum species in the American continent is discussed in view of available morphological and molecular data. A lectotype of A. mordax is here designated, and Austrodiplostomum ostrowskiae is considered as a new synonym of A. compactum.

Use of ionizing radiation in the development of vaccines against Fasciola gigantica and Schistosoma bovis in Sudanese cattle, sheep and goats

International Nuclear Information System (INIS)

Haroun, E.M.; Yagi, A.I.; Younis, S.A.; El Sanhouri, A.A.; Gadir, H.A.; Gameel, A.A.; Bushara, H.O.

1988-01-01

A primary infection with Fasciola gigantica, which had been eliminated by anthelmintic treatment after eight weeks, stimulated partial resistance to homologous challenge in calves and goats. Similar sensitization with F. gigantica or Schistosoma bovis also stimulated significant resistance to heterologous challenge in calves. The level of resistance was indicated by reduced fluke recoveries from sensitized animals compared with controls, by lower values of the enzymes sorbitol dehydrogenase and glutamate dehydrogenase, indicating less hepatic damage, and by lower blood indices, indicating more severe anaemia. Infection of calves with metacercariae of F. gigantica irradiated at 30 Gy or 200 Gy resulted in mean fluke recoveries of 75 and 41 respectively, whereas 332 flukes were recovered from controls infected with non-irradiated metacercariae. The flukes recovered from animals infected with irradiated metacercariae were stunted and resided in the small bile ductules. Sensitization for eight weeks with metacercariae of F. gigantica irradiated at 30 Gy resulted in significant resistance in calves, sheep and goats to homologous challenge. Significant resistance was also stimulated in calves by sensitization for eight weeks with metacercariae of F. gigantica irradiated at 200 Gy. However, sensitization for four weeks with F. gigantica irradiated at 30 Gy failed to stimulate significant resistance in calves to homologous challenge, but similar sensitization, twice at an interval of four weeks, resulted in an average reduction of 62% in fluke recovery from calves with homologous challenge. This reduction was, however, statistically insignificant. Moreover, sensitization for eight weeks with cercariae of Schistosoma bovis irradiated at 30 Gy did not stimulate heterologous resistance in calves or goats to F. gigantica. A similar result was also obtained after sensitization of calves for 36 weeks with cercariae of S. bovis irradiated at 30 Gy before challenge with F

Consequence of temperature changes on cercarial shedding from Galba truncatula infected with Fasciola hepatica or Paramphistomum daubneyi

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Rondelaud Daniel

2013-01-01

Full Text Available Experimental infections of Galba truncatula (two populations with Fasciola hepatica or Paramphistomum daubneyi were carried out to study the effect of water temperature changes (3 h at a mean of 12 °C every week on cercarial shedding during the patent period. The results were compared with those of control snails infected according to the same protocol and always maintained at 20 °C. Compared to controls, a significant increase in the number of cercariae-shedding snails, a significantly longer patent period and significantly greater cercarial production were noted in temperature-challenged snails, regardless of the type of digenean infection. In contrast, the number of incompletely formed metacercariae was significantly higher in temperature-challenged snails than in controls. Incompletely formed metacercariae of F. hepatica consisted of cysts whose colour remained whitish after shedding (25.4% for temperature-challenged snails or whose dome was flattened after encystment (74.6%. Those of P. daubneyi were totally dark brown or blackish after formation. These incomplete metacercariae might originate from young differentiating cercariae within the snail body (F. hepatica or from cercariae which died just after encystment (P. daubneyi. The use of regular temperature changes for snails infected with F. hepatica or P. daubneyi must be monitored carefully during collection of metacercariae to select completely formed cysts for infecting definitive hosts.

[Contribution to the study of Microphallidae Travassos 1920 (trematoda). XXXII. Microphallus breviatus n. sp., a species with an abbreviated evolutive cycle from a Mediterranean pond in the Languedoc].

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Deblock, S; Maillard, C

1975-01-01

Contribution to the study of Microphallidae Travassos, 1920 (Trematoda). XXXII. - Microphallus breviatus n. sp., a short life-cycle species of a mediterranean pond of Languedoc. The whole larval life-cycle of M. breviatus takes place in one host, Hydrobia ventrosa (Montagu), Mollusc Hydrobiidae. Hepato-pancreatic sporocysts produce morphologically altered xiphidio-cercariae which become encysted metacercariae in the sporocyts themselves. This species is defined in the genus by the anatomic characteristics of its cercariae and metacercariae, allied with its uncommon biology including two hosts only.

Import of exotic and zoonotic trematodes (Heterophyidae: Centrocestus sp.) in Xiphophorus maculatus

DEFF Research Database (Denmark)

Mehrdana, Foojan; Jensen, Hannah M.; Kania, Per Walter

2014-01-01

metacercaria with an X-shaped excretory bladder. PCR amplification of a rDNA region (5.8S rRNA gene, ITS-2, 28S rRNA gene) and subsequent sequencing confirmed the diagnosis. Metacercariae were found in gill filaments adjacent to the cartilage associated with cartilage hypertrophy, epithelial and mucous cell...... hyperplasia, clubbing and lamellar fusion. Host cell encapsulation of cysts comprised several layers of leucocytes, chondroblast-like and fibroblast like cells. The observations raise concerns with regard to veterinary inspection and quarantine procedures. The zoonotic potential of these trematodes...

Abandoning the ship: spontaneous mass exodus of Clinostomum complanatum (Rudolphi, 1814) progenetic metecercariae from the dying intermediate host Trichogaster fasciatus (Bloch & Schneider, 1801).

Science.gov (United States)

Rizvi, Asim; Alam, Md Maroof; Parveen, Saltanat; Saleemuddin, M; Abidi, S M A

2012-04-01

The dramatic and spontaneous exodus of live Clinostomum complanatum progenetic metacercaria from the gill slits of the dying intermediate host, Trichogaster fasciatus is reported. Basic water parameter tests for dissolved oxygen, pH and temperature revealed slightly lower level of dissolved oxygen in tank water used for water change. To the best of our knowledge, it is the first report of a digenean metacercariae, en mass leaving their intermediate host, upon its death in search of an alternative host to support their survival and help in continuing their life cycle.

Molecular variation in the Paragonimus heterotremus complex in Thailand and Myanmar.

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Sanpool, Oranuch; Intapan, Pewpan M; Thanchomnang, Tongjit; Janwan, Penchom; Nawa, Yukifumi; Blair, David; Maleewong, Wanchai

2013-12-01

Paragonimiasis is an important food-borne parasitic zoonosis caused by infection with lung flukes of the genus Paragonimus. Of the 7 members of the genus known in Thailand until recently, only P. heterotremus has been confirmed as causing human disease. An 8th species, P. pseudoheterotremus, has recently been proposed from Thailand, and has been found in humans. Molecular data place this species as a sister species to P. heterotremus, and it is likely that P. pseudoheterotremus is not specifically distinct from P. heterotremus. In this study, we collected metacercariae of both nominal species (identification based on metacercarial morphology) from freshwater crabs from Phetchabun Province in northern Thailand, Saraburi Province in central Thailand, and Surat Thani Province in southern Thailand. In addition, we purchased freshwater crabs imported from Myanmar at Myawaddy Province, western Thailand, close to the Myanmar-Thailand border. The DNAs extracted from excysted metacercariae were PCR-amplified and sequenced for ITS2 and cox1 genes. The ITS2 sequences were nearly identical among all samples (99-100%). Phylogenies inferred from all available partial cox1 sequences contained several clusters. Sequences from Indian P. heterotremus formed a sister group to sequences from P. pseudoheterotremus-type metacercariae. Sequences of P. heterotremus from Thailand, Vietnam, and China formed a separate distinct clade. One metacercaria from Phitsanulok Province was distinct from all others. There is clearly considerable genetic variation in the P. heterotremus complex in Thailand and the form referred to as P. pseudoheterotremus is widely distributed in Thailand and the Thai-Myanmar border region.

Experimental life history and biological characteristics of Fasciola gigantica (Digenea: Fasciolidae).

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Phalee, Anawat; Wongsawad, Chalobol; Rojanapaibul, Amnat; Chai, Jong-Yil

2015-02-01

This study was conducted to investigate the life history, morphology, and maturation of larval stages and adult worms of Fasciola gigantica in experimental mice. Lymnaea auricularia rubiginosa was used as the intermediate host, and Oryza sativa was used for encystment of the metacercariae, while Mus musculus was used as the definitive host for maturation study. Fresh eggs from the gall bladder of water buffaloes fully developed into embryonated ones and hatched out at days 11-12 after incubation at about 29ºC. Free-swimming miracidia rapidly penetrated into the snail host, and gradually developed into the next larval stages; sporocyst, redia, and daughter redia with cercariae. Fully-developed cercariae were separated from the redia and shed from the snails on day 39 post-infection (PI). Free-swimming cercariae were immediately allowed to adhere to rice plants, and capsules were constructed to protect metacercariae on rice plants. Juvenile worms were detected in intestines of mice at days 3 and 6 PI, but they were found in the bile duct from day 9 PI. Juvenile and adult flukes were recovered from 16 mice experimentally infected with metacercariae, with the average recovery rate of 35.8%. Sexually mature adult flukes were recovered from day 42 PI. It could be confirmed that experimentally encysted metacercariae could infect and develop to maturity in the experimental host. The present study reports for the first time the complete life history of F. gigantica by an experimental study in Thailand. The obtained information can be used as a guide for prevention, elimination, and treatment of F. gigantica at environment and in other hosts.

Description and proposed life cycle of Maritrema novaezealandensis n. sp. (Microphallidae) parasitic in red-billed gulls, Larus novaehollandiae scopulinus, from Otago Harbor, South Island, New Zealand.

Science.gov (United States)

Martorelli, Sergio R; Fredensborg, Brian L; Mouritsen, Kim N; Poulin, Robert

2004-04-01

Maritrema novaezealandensis n. sp. is described from Otago Harbor, South Island, New Zealand, on the basis of adult specimens collected from the Red-billed gull, Larus novaehollandiae scopulinus, and excysted metacercariae obtained from crabs. It belongs to the "eroliae group" and differs from other related species mainly in the shape, size, and patterns of distributions of the spines on the cirrus, the shape of the metraterm, the presence of an unlobed ovary, and the complete ring of the vitelline follicles. Based on morphometric features of metacercariae and adult specimens, the trophic relationships among invertebrate and vertebrate hosts, experimental infections, and previous reports of species of Maritrema with similar transmission patterns, the life cycle of M. novaezealandensis n. sp. is described. A 3-host life cycle is proposed for this parasite. The first intermediate host is the mud snail, Zeacumantus subcarinatus, in which the cercarial stage is produced in sporocysts located within the gonad of the snail. At least 3 crab species (Hemigrapsus crenulatus, Macrophtalmus hirtipes, and Halicarcinus whitei) and several species of amphipods act as second intermediate hosts, with metacercariae encysted in the body cavity of the crustacean host. Finally, the definitive host, the gull, L. n. scopulinus, harbors the adult worms in its intestine.

Microphallids in Gammarus insensibilis Stock, 1966 from a Black Sea lagoon: host response to infection.

Science.gov (United States)

Kostadinova, A; Mavrodieva, R S

2005-09-01

We examined the patterns of parasite melanization in Gammarus insensibilis using data on microphallids from Pomorie Lagoon (Black Sea) in the light of 3 predictions associated with host survival: (i) hosts invest more in defence in an environment where the likelihood for infection is higher; (ii) multiple immune challenges exhaust host reserves and result in decreased melanization rates in older hosts; (iii) host immune response is directed against the cerebral metacercariae of Microphallus papillorobustus that alter amphipod behaviour and are most detrimental to the host. G. insensibilis was capable of melanizing the metacercariae of all four species of trematodes found to be hosted by the amphipods. The frequency of melanization and mean abundance of melanized metacercariae were substantially higher than those observed in the same amphipod-gammarid system on the French Mediterranean coast. However, the rate of melanization was low and showed a significant decrease with amphipod size. Although the 4 species were differentially melanized, the host response was largely directed against Microphallus hoffmanni and M. subdolum. We suggest that (i) the lower melanization efficiency with age is due to the mode of infection, probably leading to loss of haemolymph and monopolization of the defence resources for wound healing and (ii) in the French system, host response focuses on the most prevalent and abundant species.

The life cycle of Ascocotyle (Phagicola) longa (Digenea: Heterophyidae), a causative agent of fish-borne trematodosis.

Science.gov (United States)

Simões, Susana Balmant Emerique; Barbosa, Helene Santos; Santos, Cláudia Portes

2010-03-01

The complete life cycle of the trematode Ascocotyle (Phagicola) longa (Digenea: Heterophyidae) is elucidated by natural observation validated by experimental infections. The natural first intermediate host of A. (P.) longa, an agent of human heterophyiasis in Brazil, is the cochliopid snail Heleobia australis (new first intermediate host). Metacercariae were found encysted in the body musculature, heart, stomach, liver, kidney, spleen, gonads and mesentery of mullets Mugil liza. Hamsters Mesocricetus auratus were experimentally infected with metacercariae of A. (P.) longa obtained from the mullets, and the adults recovered were used to infect the snails H. australis. Rediae and cercariae of A. (P.) longa are described for the first time. The ultrastructure of the tegument of A. (P.) longa shows a change in spination pattern from the cercaria with single-pointed spines to the metacercaria and adult with multipointed, brush-shaped spines. The life cycle of A. (P.) longa is related to estuaries and coastal lagoons where the recruitment of mugilid juveniles occurs. The high prevalence (100%) of A. (P.) longa encysted in the mullets examined within the urban area of Rio de Janeiro indicates the potentially great public health impact of the consumption of raw mullets. Copyright 2009 Elsevier B.V. All rights reserved.

The life history of Pleurogenoides malampuzhensis sp. nov. (Digenea: Pleurogenidae) from amphibious and aquatic hosts in Kerala, India.

Science.gov (United States)

Brinesh, R; Janardanan, K P

2014-06-01

The life-cycle stages of Pleurogenoides malampuzhensis sp. nov. infecting the Indian bullfrog Hoplobatrachus tigerinus (Daudin) and the skipper frog Euphlyctis cyanophlyctis (Schneider) occurring in irrigation canals and paddy fields in Malampuzha, which forms part of the district of Palakkad, Kerala, are described. The species is described, its systematic position discussed and compared with the related species, P. gastroporus (Luhe, 1901) and P. orientalis (Srivastava, 1934). The life-cycle stages, from cercaria to egg-producing adult, were successfully established in the laboratory. Virgulate xiphidiocercariae emerged from the snail Digoniostoma pulchella (Benson). Metacercariae are found in muscle tissues of dragonfly nymphs and become infective to the frogs within 22 days. The pre-patent period is 20 days. Growth and development of both metacercariae and adults are described.

Experimental Life Cycle of Hypoderaeum conoideum (Block, 1872 Diez, 1909(Trematoda: Echinostomatidae Parasite from the North of Iran

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Hakim AZIZI

2015-03-01

Full Text Available Background: Human Echinostomiasis is an intestinal disease caused by the members of family Echinostomatidae parasites. The aim of present research was to identify echinos­tomatidae cercariae emitted by Lymnaea palustris snails from Mazandaran province in the north of Iran based on the morphological and morphometrical charac­teristics of the different stages of experimental parasite life cycle.Methods: Echinostomatidae cercariae were collected from L. palustris (Gastropoda: Lymnaeidae of the north of Iran. To collect metacercaria, 50 healthy snails were infected with cercariae experimentally (50 cercariae for each. To obtain the adult stage, 9 laboratory animals (3 ducks, 2 rats, 2 mice and 2 quails were fed with 60 metacercaria for each. To identify parasite, the different stages of worm were exam­ined using light microscope and then the figures were draw under camera Lucida microscope and measures were determined.Results: Averagely, 15metacercaria were obtained from each snail that had been previously exposed with cercariae. Ducks presented worm eggs in feces after 10-15 days post-infection. Intestinal worms were collected and identified as Hypoderaeum conoideum on the bases of figures and measures of cephalic collar, the number of collar spine, suckers diameter ratio, testes arrangement, etc.Conclusion: H. conoideum cercariae and adult worm are described. This is the first report of the different stages of the experimental life cycle of this parasite in Iran.

A new species of Microphallus (Trematoda: Microphallidae from Venezuela

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M.T Díaz

2004-06-01

Full Text Available During 1997-1999, a total of 94 crabs, Uca rapax were collected from La Sabana, La Ceiba and El Paujil, Sucre State, Venezuela. Of these 36 were infected with metacercariae. Two parasites were located in the abdominal muscles and one under the tissue of carapace and gonad. These metacercariae grew to adults in the following genera: Levinseniella, Microphallus and Maritrema, in the period of 2-5 days after feeding experimentally to the rat Rattus norvegicus, mice Mus musculus and duck Cairinia moschata. Specimens of the genus Microphallus were described herein as a new species M. sabanensis. The life cycle of M. sabanensis sp.nov. were studied experimentally using rat, mice and duck. All developmental stages and the adult are described. In addition, M. sabanensis was collected from wild birds Anas discors, Pluvialis squatarola, Butorides striatus, Egretta caerulea and Nycticorax violaceus from the same localitiesDurante 1997-1999, se recolectaron en La Sabana, La Ceiba y El Paujil del Distrito Cajigal, Estado Sucre, Venezuela, un total de 94 cangrejos Uca rapax, 36 de ellos estaban parasitados con tres especies de metacercarias de digéneos. Dos de ellos localizados en la musculatura del abdomen y una en el tejido que recubre internamente el caparazón y las gónadas. Estas metacercarias fueron dadas como alimento a ratones Mus musculus, ratas Rattus norvegicus, y patos Cairina moschata y tres genéros de microfálidos, Levinseniella, Microphallus y Maritrema fueron recuperados del intestino delgado de estos animales después de 2-5 días de la infección. Especimenes del genero Microphallus de este estudio se describen como Microphallus sabanensis sp.n. Se estudió experimentalmente los aspectos de ciclo vida de la especie, utilizando ratas, ratones y patos. Se describen todas las estapas larvales y los adultos. Además, M. sabanensis se encuentra naturalmente en las aves: Anas discors, Pluvialis squatarola, Butorides striatus, Egretta caerulea

The effect on infection of irradiated fasciola gigantic (liver flukes) on goats (Capra hircus linn) blood values

International Nuclear Information System (INIS)

Tuasikal, BJ.; M-Arifin; Yusneti; Dinardi; Suhardono

2006-01-01

This experiment was carried out to study the effect of infection Fasciola gigantic irradiated by using 60 Co to goat’s blood values. Twenty local male goats, ten to twelve old months, 115-20 kg of body weight were used for this study. The animals were divided into 5 groups consisting of four replications. Three groups received irradiated metacercariae of Fasciola gigantic a using 45, 55 and 65 Gy respectively. One group in the positive control and another one is the negative control. Each goat received 350 metacercariae. The study in was undertaken for 25 weeks. Body weight, Eosinofil, PCV and Hb were measured. These indicated that infectivity reduced on irradiated Fasciola gigantic. In all parameters, among negative control group and three irradiation-groups showed non significant (p>0,05). (author)

prevalence of bovine fasciolosis in major abattiors of adamawa state

African Journals Online (AJOL)

DR. AMINU

the metacercariae during grazing on vegetation. Tropical ... January-March when relative humidity drops to 13%. Mubi ... at Yola abattoir; ten (10) animals were examined in. Mubi, while ..... Soulsby, E. J. W. (1982): Helminths, Arthropods and.

Studies on some fish parasites of public health importance in the southern area of Saudi Arabia

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Mokhtar Ibrahim Khalil

2014-12-01

Full Text Available The present study was the first attempt to survey the diversity of fish zoonotic parasites in the southern region of Saudi Arabia, particularly the Najran area, from October 2012 to October 2013. Approximately 163 fish representing seven species (two of freshwater fish and five of marine fish were examined for fish-borne trematode metacercariae using the compression technique, and for zoonotic nematode larvae. Adult flukes were obtained from cats experimentally infected with the metacercariae on day 25 post-infection The prevalence of each parasite species was recorded. The parasites found belonged to two taxa: Digenea (Heterophyes heterophyes and Haplorchis pumilio in muscle tissue; and nematodes (larvae of Capillaria sp. in the digestive tract. The morphological characteristics of the fish-borne trematode metacercariae and their experimentally obtained adults were described. This is the first report of these parasites in fish in Saudi Arabia. Moreover, Myripristis murdjan presented higher prevalence of Capillaria sp. infection (22.7%, while Haplorchis pumilio was the dominant metacercarial species (7.9%. Although the number of documented cases continues to increase, the overall risk of human infection is slight. The increasing exploitation of the marine environment by humans and the tendency to reduce cooking times when preparing seafood products both increase the chances of becoming infected with these parasites. Furthermore, our results indicate that certain fish production systems are at risk of presenting fish zoonotic parasites, and that control approaches will benefit from understanding these risk factors.

Studies on some fish parasites of public health importance in the southern area of Saudi Arabia.

Science.gov (United States)

Khalil, Mokhtar Ibrahim; El-Shahawy, Ismail Saad; Abdelkader, Hussein Saad

2014-01-01

The present study was the first attempt to survey the diversity of fish zoonotic parasites in the southern region of Saudi Arabia, particularly the Najran area, from October 2012 to October 2013. Approximately 163 fish representing seven species (two of freshwater fish and five of marine fish) were examined for fish-borne trematode metacercariae using the compression technique, and for zoonotic nematode larvae. Adult flukes were obtained from cats experimentally infected with the metacercariae on day 25 post-infection The prevalence of each parasite species was recorded. The parasites found belonged to two taxa: Digenea (Heterophyes heterophyes and Haplorchis pumilio) in muscle tissue; and nematodes (larvae of Capillaria sp.) in the digestive tract. The morphological characteristics of the fish-borne trematode metacercariae and their experimentally obtained adults were described. This is the first report of these parasites in fish in Saudi Arabia. Moreover, Myripristis murdjan presented higher prevalence of Capillaria sp. infection (22.7%), while Haplorchis pumilio was the dominant metacercarial species (7.9%). Although the number of documented cases continues to increase, the overall risk of human infection is slight. The increasing exploitation of the marine environment by humans and the tendency to reduce cooking times when preparing seafood products both increase the chances of becoming infected with these parasites. Furthermore, our results indicate that certain fish production systems are at risk of presenting fish zoonotic parasites, and that control approaches will benefit from understanding these risk factors.

In vitro cultivation of Maritrema novaezealandensis (Microphallidae)

DEFF Research Database (Denmark)

Fredensborg, Brian Lund; Poulin, R

2005-01-01

In this study, the effect of the composition of culture medium on excystation, growth, survival and egg production was investigated for the recently discovered microphallid trematode Maritrema novaezealandensis. Metacercariae from the two second intermediate crab hosts, Macrophthalmus hirtipes an...

Scanning electron microscopy of Clinostomum metacercaria from ...

African Journals Online (AJOL)

Egyptian Journal of Biology ... At the ventral side, the rims of the oral and ventral suckers had minor differences, while their floor showed great variation in topography: the floor of the oral sucker had longitudinal wrinkles with nodule-like structures and their upper margins had minute sensory papillae, while the ventral sucker ...

Eye-flukes in fish, living in cooling water from nuclear power plants

International Nuclear Information System (INIS)

Hoeglund, J.; Thulin, J.

1988-01-01

We report here on the effects of raised water temperature on the prevalence, mean infrapopulation density and consequences of eye-flukes in fish. The study was mainly performed in the Biotest basin situated 120 km north of Stockholm, Sweden. This 1 km 2 basin is an enclosed brackish water (5 permillage) area receiving heated (about 8 degree C) cooling water (90 m 3 /s) from Forsmark nuclear power station. Both morphological and experimental studies of the parasite larvae of sampled fish indicate that we are dealing with four strains of Diplostomum, two of which occur in perch and the other two in roach. Since taxonomic revisions are under hand elsewhere we prefer to name these strains Diplostomum sp1-4. Metacercariae of D.sp 1 were found between the retina and sclera in the eye of perch while that of D.sp 4 were found in the eye-lens of roach in over 90 % of fish examined. Metacercariae of the other two D.-species and of Tylodelphys clavata and Cotylurus sp were found at lower frequencies. Cercariae of Diplostomum spp were found to develop from sporocysts in snails of the genus Lymnaea. The period of cercarial shedding starts about one month earlier and is also prolonged in the Biotest basin compared to the reference locality. The infection procedure, however, is the same in both areas. During experimental infections with cercariae on yearlings of bleak we found a distinct correlation between an increased fry mortality and an increased cercariae density, a connection which was strengthened at increased water temperature. Furthermore, the results indicate that the defence mechanisms of the fish respond slower towards infections with Diplostomum spp than that is known to be the case with bacterial infections. The speed with which the metacercariae accumulate in the eye of the fish is higher in the Biotest basin than in the reference locality. In spite of this, the mean infrapopulation density of metacercariae in older fish is not higher here than in the reference

The prevalence of Proctoeces (Trematoda: Fellodistomidae ...

African Journals Online (AJOL)

1988-05-03

May 3, 1988 ... between sex of host and parasitic infection, with female mussels harbouring the most metacercariae. ... possible pathogenic effects of Proctoeces on such highly stressed mussel populations clearly ... Histological studies of the reproductive cycle of the ..... variations in transmission success are undoubtedly.

Helminth parasites of Xenotaenia resolanae (Osteichthyes: Cyprinodontiformes: Goodeidae) from the Cuzalapa hydrological system, Jalisco, Mexico.

Science.gov (United States)

Martínez-Aquino, Andrés; Aguilar-Aguilar, Rogelio; Pérez-Rodríguez, Rodolfo; de León, Gerardo Pérez-Ponce

2009-10-01

Six helminth species were recorded during the helminthological examination of 35 specimens of the goodeid Xenotaenia resolanae from Arroyo Durazno, Jalisco, Mexico, a tributary of the Cuzalapa River. Helminth species identified included: 4 species of digeneans, i.e., Posthodiplostomum minimum (metacercariae), Clinostomum companatum (metacercariae), Dendrorchis sp. (adult), and Margotrema guillerminae (adult); and 2 species of nematodes, i.e., Spiroxys sp. (larvae) and Rhabdochona ahuehuellensis (adult). A very low number of individual larvae were found. The observed species richness, individual parasite abundance, and diversity were low at both component community and infracommunity levels. The values of similarity between infracommunities were relatively high because of the predominance of the digenean M. guillerminae, the species that reached the higher values of both prevalence and abundance. High water flow of the collecting site is suggested as the main factor determining the depauperate helminth assemblage in this fish species.

[Epidemiological investigation of second intermediate hosts of Clonorchis sinensis in Pizhou and Xinyi cities of Jiangsu Province].

Science.gov (United States)

Jin, Xiao-lin; Liu, Jian-feng; Shen, Ming-xue; Dai, Yang; Xu, Xiang-zhen

2015-12-01

To understand the endemic situation of Clonorchis sinensis in its second intermediate hosts in Pizhou and Xinyi cities of Jiangsu Province, so as to provide the evidence for the further control and treatment. Pseudorasbora parva and Abbottina rivularis were caught from the natural water body of Pizhou and Xinyi cities, and the tabletting microscopy method was applied to test the metacercaria of C. sinensis. Totally 1 117 fishes were caught and dissected, including 792 P. parva (70.90%) and 325 A. rivularis (29.10%). The metacercaria of C. sinensis infection rates of P. parva and A. rivularis were 29.80% (236/792) and 4.62% (15/325) respectively, and the difference between the two kinds of fishes was significant (χ² = 83.88, P sinensis infection rate of freshwater fishes in Pizhou and Xinyi cities is high, and the local residents are facing the higher risk of clonorchiasis sinensis.

Second intermediate host land snails and definitive host animals of Brachylaima cribbi in southern Australia

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Butcher A.R.

2005-03-01

Full Text Available This study of infection of southern Australian land snails with Brachylaima cribbi metacercariae has shown that all commonly encountered native and introduced snails are susceptible second intermediate hosts. The range of infected snails is extensive with metacercariae-infected snails being present in all districts across southern Australia. C. virgata has the highest average natural metacercarial infection intensity of 6.1 metacercariae per infected snail. The susceptibility of birds, mammals and reptiles to B. cribbi infection was studied in South Australia by capturing, dissecting and examining the intestinal tract contents of animals which commonly eat land snails as a food source. Indigenous Australian little ravens (Corvus mellori, which are a common scavenger bird, and two other passeriform birds, the black bird (Turdus merula and the starling (Sturnus vulgaris, which are both introduced European birds, were found to have the highest infection rates of all animals examined. Other birds found infected with B. cribbi were an emu (Dromaius novaehollandiae, chickens (Gallus gallus and a pigeon (Columba livia. Natural infections were also detected in field mice (Mus domesticus and shingleback lizards (Tiliqua rugosa although the intensity of infection was lower than that observed in birds. Susceptibility studies of laboratory mice, rats and ducks showed that mice developed patent infections which persisted for several weeks, rats developed a short-lived infection of three weeks’ duration and ducks did not support infection. This study has shown for the first time that a brachylaimid can infect a wide host range of birds, mammals and reptiles in nature.

Using mitochondrial and ribosomal DNA sequences to test the taxonomic validity of Clinostomum complanatum Rudolphi, 1814 in fish-eating birds and freshwater fishes in Mexico, with the description of a new species.

Science.gov (United States)

Sereno-Uribe, Ana L; Pinacho-Pinacho, Carlos D; García-Varela, Martín; de León, Gerardo Pérez-Ponce

2013-08-01

The taxonomic history and species composition of the genus Clinostomum has been unstable. Two species, Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819, have been particularly problematic and its validity has been disputed for nearly 200 years. In this paper, we have made use of an integrative taxonomy approach, and we used, in first instance, DNA sequences of two genes (cox1 and ITS) to test the validity of C. complanatum, a species apparently widely distributed in Mexico and to link the metacercariae and adult forms of the recognized species of Clinostomum. Combining molecular data with morphology, host association, and geographical distribution, we searched for the potential existence of undescribed species. A new species of Clinostomum is described based on adults found in the mouthy cavity of three species of fish-eating birds as well as in metacercariae found in freshwater and estuarine fishes. A few morphological characteristics distinguish the new species from other congeners even though reciprocal monophyly in a phylogenetic tree based on maximum-likelihood and Bayesian analysis, genetic divergence, and a multivariate analysis of variance and a principal component analysis of 18 morphometric traits for adults and metacercariae demonstrates the validity of the new species. Based on our results, it seems that C. complanatum is not currently distributed in Mexico, although this requires further verification with a more thoroughful sampling in other areas of the country, but it is plausible to support the hypothesis that C. marginatum is the American form, as previously suggested by other authors.

Experimental evidence for parasite-induced over-winter mortality in juvenile Rhodeus amarus

Czech Academy of Sciences Publication Activity Database

Michálková, Veronika; Ondračková, Markéta

2014-01-01

Roč. 84, č. 5 (2014), s. 1377-1388 ISSN 0022-1112 R&D Projects: GA ČR GBP505/12/G112 Institutional support: RVO:68081766 Keywords : Bitterling * Diplostomum pseudospathaceum * Metacercariae * Parasite infection * Trematoda Subject RIV: EG - Zoology Impact factor: 1.658, year: 2014

the assessment of fasciola gigantica infection in the rabbit ...

African Journals Online (AJOL)

In this study, the rabbit was assessed as a laboratory host for the helminthes parasite, Fasciola gigantica. Three groups of rabbits were Infected experimentally with 5, 10 end 15 metacercariae of F. gigantica respectively. Clinical signs found included pale mucous membrane, progressive emaciation and rough hair coat.

Effect of fish size on transmission of fish-borne trematodes (Heterophyidae) to common carps (Cyprinus carpio) and implications for intervention

NARCIS (Netherlands)

Boerlage, A.S.; Graat, E.A.M.; Verreth, J.A.J.; Jong, de M.C.M.

2011-01-01

Fish-borne trematodes are reported to affect the health of more than 40 million people worldwide. Few experimental studies are available on fish size dependent gain (attack rates of cercariae) or loss (mortality of metacercariae) of fish-borne trematodes. Aim was to quantify the relation between

Temporal variation in the infection dynamics and maturation cycle of Oligogonotylus manteri (Digenea) in the cichlid fish, 'Cichlasoma' urophthalmus, from Yucatán, México.

Science.gov (United States)

Jiménez-García, M I; Vidal-Martínez, V M

2005-10-01

We studied the infection dynamics and maturation cycle of Oligogonotylus manteri in wild and caged 'Cichlasoma' urophthalmus, and determined the potential role of different sources of infection in its transmission in a quarry (MITZA). Metacercariae, and nongravid and gravid stages of O. manteri were present throughout 1 annual cycle. Prevalence, mean intensity, and/or aggregation values peaked around April and June in both wild and caged fish. This period of time includes the start of the rainy season, in which the water temperature reaches its maximum annual values. Because temperature is a major factor triggering 'C.' urophthalmus activity (food intake, growth, and reproduction), and O. manteri metacercariae and adults are trophically transmitted, temperature may be playing an important role in the recruitment of worms to the fish. We also determined that cercariae infect caged fish through a mechanism other than trophic transmission whereby fish consume infected snails, which has been described as the most common mode of transmission to 'C.' urophthalmus.

Posthodiplostomatosis in a fishpond in Serbia

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Marković Maja

2012-01-01

Full Text Available Posthodiplostomatosis (black spot disease is a disease of young freshwater fish species of families Cyprinidae and Cobitidae, resulting from infection with metacercaria, which is the penultimate developmental stage of Posthodiplostomum cuticola. Metacercaria give rise to black cysts located primarily in the skin, muscles and on the fins. The disease caused by P. cuticola in certain fish species is first detected in a fishpond in Serbia, as presented in the current work, whilst in natural waters it has been existing for many years. Diagnosis of the infection in the grass carp (Ctenopharyngodon idella and the bighead carp (Aristichthys nobilis was based on the occurrence of dark discolorations and cysts of the parasite on the fins. Cysts measured approximately 1 mm in diameter, were confirmed by histopathological means. Posthodiplostomatosis occurred in fishponds in which preventive measures were not implemented, i.e. where the nurseries were not dried up and mechanically cleaned.

Development of nuclear techniques in food and life engineering - Irradiation of food and drinking water to control parasitic infections

Energy Technology Data Exchange (ETDEWEB)

Chai, Jong; Shin, Eun Hee; Guk, Sang Mee; Han, Eun Taek; Kim, Jae Lip [Seoul National University, Seoul (Korea)

2000-04-01

To prevent some intestinal trematodes which are highly prevalent in Korea, the effect of irradiation on fishes and metacercariae has been analyzed for control of parasites. The metacercariae of intestinal trematodes were irradiated with various doses, and the infection rates and developmental status were assayed for estimation of optimal radiation doses. In addition, to examine the differences of susceptibility to radiation among the parasites, superoxide dismutase (SOD) activities were determined, and degree of apoptosis were observed. As a result, 100 Gy irradiation was sufficient for Metagonimus yokogawai to redue their infectivity and development, and 200 Gy was for Gymnophalloides seoi. While Neodiplostomum seoulense, a kind of intestinal rematode, was proved to be highly susceptible to radiation, sparganum and Anisakis were resistant to radiation. These might result from the differences of SOD activity among parasites. The apoptosis of parasites was mainly observed after 100 Gy irradiation, and further study was required to decide whether these phenomenon influences the susceptibility to radiation. 21 refs., 7 figs., 4 tabs. (Author)

Raw-fish-eating behavior and fishborne zoonotic trematode infection in people of northern Vietnam

DEFF Research Database (Denmark)

Phan, Van Thi; Ersbæll, Annette Kjær; Do, Dung Trung

2011-01-01

, as they know effective drug treatment is available. Raw fish dishes are consumed at social gatherings from shared plates and dipping sauces using the same chop sticks. This is likely to pose risks of crosscontamination with FZT metacercariae to different food items as indicated by the finding that 25...

Some parasitic worms in freshwater fishes and fish-predators from ...

African Journals Online (AJOL)

This report concerns a collection of parasitic worms from South Africa, and contains a new species of monogenean, Gyrodactylus transvaalensis, six species of adult trematodes, including a new species of Phyllodistomum.P. van der waali, two metacercariae of the Family Clinostomidae and three of the Order Strigeida, ...

Fasciola gigantica transmission in the zoonotic fascioliasis endemic lowlands of Guilan, Iran: experimental assessment.

Science.gov (United States)

Ashrafi, Keyhan; Mas-Coma, Santiago

2014-09-15

The lowland flatlands around the city of Bandar-Anzali, at the Caspian Sea shore, Guilan province, are an endemic area where Fasciola gigantica appears to be the fasciolid species involved and past outbreaks affecting around 15,000 people and the highest human infection rates in Iran have been reported. Fascioliasis transmission in that area has been experimentally analysed for the first time, by means of assays of monomiracidial (Group A: 120 snails) and pentamiracidial (Group B: 96 snails) infections of local Radix lymnaeid snails with a local cattle F. gigantica isolate. Ribosomal DNA ITS-2 sequencing proved that Lymnaea (Radix) gedrosiana should henceforth be considered a synonym of Radix auricularia, the haplotype found in Bandar-Anzali being identical to that found in many European countries. Survival rates at day 30 postinfection and metacercarial productivity (both higher in Group A) and longevity of the shedding snails (higher in Group B), were affected by the miracidial dose, whereas prepatent period, patent period, infection rate, and the percentages of shedding and infected non-shedding snails did not. The higher percentage of shedding snails in Group A (51.0% versus 37.7%) counteracts the higher number of metacercariae produced in Group B (243.9 ± 259.2 versus 157.2 ± 153.2). High numbers of shedding snails in both experimental groups passed less than 100 cercariae, and 16% in Group A and 35% in Group B produced more than 300 metacercariae, while only four snails (8%) in Group A and two snails (10%) in Group B shed more than 500 metacercariae. Most metacercariae (94.7% in Group A and 85.1% in Group B) were recorded during the first 15 days of patent period. The comparison with results from other fasciolid/lymnaeid systems indicates that the F. gigantica/R. auricularia system of Guilan is highly susceptible and compatible. Results obtained suggest that increased lymnaeid vector populations and not polymiracidial snail infections most probably underlay

A macro- and light microscopical study of the pathology of Calicophoron microbothrium infection in experimentally infected cattle

Directory of Open Access Journals (Sweden)

M. Mavenyengwa

2005-09-01

Full Text Available Twelve Tuli weaner steers aged 1 year were randomly subdivided into three groups of four animals and infected with different doses of Calicophoron microbothrium metacercariae. Each animal in Group I received a low dose (LD of 5 000 metacercariae, Group II a medium dose (MD of 15 000 metacercariae, Group III a high dose (HD of 25 000 metacercariae and one additional animal was kept as an uninfected control (C. After infection, one animal from each group was slaughtered on Day 28, 42, 56 and 84 post infection (pi and samples from the ileum, jejunum, duodenum, abomasum and the rumen were collected for histopathological and cytological examination. On Day 28 pi, the gross pathological lesions observed in the duodenum of the LD and the MD animals were similar and comprised duodenal thickening, corrugation, hyperaemia, petechiation and ulceration. In the HD animal the duodenal lesions were similar but more severe. The abomasal folds were severely oedematous in the MD group and nearly occluded the abomasal lumen. Moderate oedema of the abomasal folds was also present in the LD and HD animals. The gross pathological lesions regressed in all the infected groups with increasing age of infection and had disappeared completely by Day 56 pi. On Day 28 pi the histopathological lesions in the duodenum and jejunum of the LD and MD groups were similar, comprising subtotal villous atrophy, hyperplasia of Brunner's glands and Peyer's patches and moderate infiltration of eosinophils, mast cells and a few globule leukocytes, basophils and lymphocytes in the lamina propria. The HD group had total villous atrophy, severe hyperplasia and cystic dilatation of Brunner's glands, which had expanded to cover the entire submucosa. On Day 42 pi the histopathological lesions were still present in the MD and the HD groups comprising subtotal villous atrophy and hyperplasia of Brunner's glands. Heavy infiltrations of eosinophils, moderate amounts of mast cells and a few

Characterization and vaccine potential of Fasciola gigantica saposin-like protein 1 (SAP-1).

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Kueakhai, Pornanan; Changklungmoa, Narin; Waseewiwat, Pinkamon; Thanasinpaiboon, Thanaporn; Cheukamud, Werachon; Chaichanasak, Pannigan; Sobhon, Prasert

2017-01-15

The recombinant Fasciola gigantica Saposin-like protien-1 (rFgSAP-1) was cloned by polymerase chain reaction (PCR) from NEJ cDNA, expressed in Escherichia coli BL21 (DE3) and used for production of a polyclonal antibody in rabbits (anti-rFgSAP-1). By immunoblotting and immunohistochemistry, rabbit IgG anti-rFgSAP-1 reacted with rFgSAP-1 at a molecular weight 12kDa, but not with rFgSAP-2. The rFgSAP-1 reacted with antisera from mouse infected with F. gigantica metacercariae collected at 2, 4, and 6 weeks after infection. The FgSAP-1 protein was expressed at a high level in the caecal epithelium of metacercariae and NEJs. The vaccination was performed in Imprinting Control Region (ICR) mice (n=10) by subcutaneous injection with 50μg of rFgSAP-1 combined with Alum adjuvant. Two weeks after the second boost, mice were infected with 15 metacercariae per mouse by the oral route. The percents protection of rFgSAP-1 vaccine were estimated to be 73.2% and 74.3% when compared with non vaccinated-infected and adjuvant-infected controls, respectively. The levels of IgG1 and IgG2a specific to rFgSAP-1 in the immune sera, which are indicative of Th2 and Th1 immune responses, were inversely and significantly correlated with the numbers of worm recoveries. The rFgSAP-1-vaccinated mice showed significantly reduced levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and liver damage. These indicated that rFgSAP-1 has strong potential as a vaccine candidate against F. gigantica, whose efficacy will be studied further in large economic animals including cattle, sheep, and goat. Copyright © 2016 Elsevier B.V. All rights reserved.

An experimental study on cerebral paragonimiasis using cats

International Nuclear Information System (INIS)

Lee, Seon Kyu; Chang, Kee Hyun; Goo, Jin Mo; Han, Moon Hee; Shin, Yong Moon; Choo, Sung Wook; Yu, In Kyu; Cho, Seung Yull; Kong, Yoon

1994-01-01

It is important to diagnosis paragonimiasis in early active because it can be dared by chemotherapy. However, it is difficult to make a correct diagnosis of cerebral paragonimiasis in the early active stage, and the radiographic findings of cerebral paragonimiasis have been rarely reported. Thus, this experimental study was designed to produce early active cerebral paragonimiasis and to demonstrate radiologic-pathologic correlations. In 8 cats, 7-8 metacercariae of Paragonimus Westermani were directly introduced into brain parenchyma of each cat's after trephination of the skull. In another 16 cats, the juvenile worms and the adult worms that had developed for varying periods (2 weeks, 4 weeks, 6 weeks, 8 weeks and 12 weeks) in the lunges of another cats were introduced into the brain parenchyma of each cat's with the same procedure described above. Follow -up MR images and chest radiographs were obtained at 2 days, 1 weeks, 2 weeks, 4 weeks and 8 weeks after inoculation. The autopsies and histopathological examinations of the cat's brain were undertaken in 22 cats. In 9 cats that were suspected with pulmonary lesion on chest radiograph, the soft tissue radiographs of inflated-fixed lungs were obtained. In one cat with inoculation of adult worm, acute suppurative inflammation of the brain parenchyma was demonstrated. But the other cats with inoculation of adult worm or juvenile worm and the cats with intentional of metacercaris did not reveal any evidence of acute cerebral paragonimiasis. More than half of the introduce metacercariae (5 out of 8 cats) were found in the lung parenchyma, while only 25% (4 out of 16 cats) of the adult worm inoculated cats were. Acute suppurative inflammation suggesting acute stage cerebral paragonimiasis was obtained in one case of adult worm inoculated cat. Most of the inoculated metacercariae and some of the juvenile worms or adult worms were migrated to the lungs

Prevalence of Clonorchis sinensis infection in freshwater fishes in northeastern China.

Science.gov (United States)

Zhang, Y; Chang, Q C; Zhang, Y; Na, L; Wang, W T; Xu, W W; Gao, D Z; Liu, Z X; Wang, C R; Zhu, X Q

2014-08-29

The prevalence of Clonorchis sinensis infection in freshwater fishes was surveyed in Heilongjiang Province, northeastern China, between August 2011 and September 2013. Thirteen species of freshwater fish (n=3221) and one species of shrimp (n=93) were collected from Songhua river, Nenjiang river and other lakes or ponds in 37 sites of 15 representative cities in Heilongjiang Province. They were individually examined by digestion technique, and the C. sinensis metacercariae were identified morphologically followed by confirmation using sequences of the second internal transcribed spacer of ribosomal DNA. Ten of the 13 examined species of freshwater fishes were infected with C. sinensis metacercariae, while all shrimps were negative. The overall prevalence of C. sinensis infection in 3221 examined freshwater fishes was 19.96%, with 42.57% (272/639) in Pseudorasbora parva, 22.55% (83/368) in Hemicculter leuciclus, 20.44% (121/592) in Carassius auratus, 17.71% (68/384) in Saurogobio dabryi, 10.85% (23/212) in Rhodeus ocellatus, 10.54% (48/455) in Phoxinus lagowskii, 8.20% (21/256) in Perccottus glehnii, 6.25% (5/80) in Misgurnus anguillicaudatus, 4.55% (1/22) in Xenocypris davidi, and 1.49% (1/67) in Cyprinus carpio. The average infection intensity in P. parva was 103.3 encysted metacercariae per gram of fish meat in Zhaoyuan city. The average prevalence of C. sinensis infection in Songhua river, Nenjiang river and lakes or ponds were 31.96% (503/1574), 11.30% (102/903) and 7.93% (59/744), respectively. The prevalence of C. sinensis infection in Zhaoyuan city (43.68%) was the highest among all sampling locations. These results revealed a high-prevalence of C. sinensis infection in freshwater fishes in Heilongjiang Province, northeastern China, posing significant public health concern. Copyright © 2014 Elsevier B.V. All rights reserved.

Molecular characterization and expression of Rab7 from Clonorchis sinensis and its potential role in autophagy.

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Jia, Feifei; Li, Ye; Huang, Yan; Chen, Tingjin; Li, Shan; Xu, Yanquan; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing

2013-07-01

Accumulating evidences suggest that Rab7 GTPase is important for the normal progression of autophagy. However, the role of Rab7 GTPase in regulation of autophagy in Clonorchis sinensis is not known. In this study, a gene encoding Rab7 was isolated from C. sinensis adult cDNA. Recombinant CsRab7 was expressed and purified from Escherichia coli. CsRab7 transcripts were detected in the cDNA of adult worm, metacercaria, cercaria, and egg of C. sinensis, and were highly expressed in the metacercaria. Immunohistochemical localization results revealed that CsRab7 was specifically deposited on the vitellarium and eggs of adult worm. Furthermore, EGFP signal of CsRab7WT and the active mutant CsRab7Q67L were associated with autophagic vesicles in transiently transfected 293T cells. It is concluded from the present study that CsRab7 GTPase possibly contributes to the development of C. sinensis and that the autophagy pathway could be an important site of action with respect to the developmental role of CsRab7 in C. sinensis.

Molecular and morphological evidence for three species of Diplostomum (Digenea: Diplostomidae), parasites of fishes and fish-eating birds in Spain

Czech Academy of Sciences Publication Activity Database

Pérez-del-Olmo, A.; Georgieva, Simona; Pula, H.J.; Kostadinova, Aneta

2014-01-01

Roč. 7, NOV 12 2014 (2014), s. 502 ISSN 1756-3305 R&D Projects: GA ČR GBP505/12/G112 Institutional support: RVO:60077344 Keywords : Diplostomum spathaceum * Diplostomum pseudospathaceum * Lens metacercariae * Freshwater fish * Gulls * Spain * Cox1 * TS1-5.8S-ITS2 Subject RIV: EG - Zoology Impact factor: 3.430, year: 2014

Sphincterodiplostomum musculosum (Digenea, Diplostomidae infecting Steindachnerina insculpta (Characiformes, Curimatidae in the Chavantes Reservoir, Southeastern Brazil Sphincterodiplostomum musculosum (Digenea, Diplostomidae infectando Steindachnerina insculpta (Characiformes, Curimatidae no reservatório de Chavantes, Sudeste do Brasil

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Aline Cristina Zago

2013-03-01

Full Text Available This study aimed to report the infection by Sphincterodiplostomum musculosum metacercariae in Steindachnerina insculpta from the Chavantes Reservoir, medium Paranapanema River, municipality of Ipaussu, State of São Paulo, Brazil. Thirty specimens of S. insculpta were collected and 29 were infected with S. musculosum metacercariae (prevalence = 96.67% in the eyes and visceral cavity. The mean intensity of infection and abundance were 96.6 ± 29.41 (7-846 and 93.3 ± 28.6 (0-846, respectively. Positive correlation was observed between parasite abundance in the eyes and standard length (rs = 0.5, p = 0.005, total weight (rs = 0.649, p = 0.0001, and condition factor (rs = 0.439, p = 0.0154. The high parasitism rates by S. musculosum metacercariae in S. insculpta can be an indicative that this fish species is highly susceptible to infection by this diplostomid, and even reflect the presence of a large abundance of the intermediate host. Moreover, S. musculosum metacercariae are reported for the first time in S. insculpta.O presente estudo tem como objetivo relatar a infecção por metacercárias de Sphincterodiplostomum musculosum em Steindachnerina insculpta provenientes do Reservatório de Chavantes, médio Paranapanema, município de Ipaussu, Estado de São Paulo, Brasil. Foram coletados 30 espécimes de S. insculpta, sendo que 29 estavam infectados por metacercárias de S. musculosum (prevalência = 96,67% nos olhos e na cavidade visceral. A intensidade média de infecção e abundância apresentaram valores de 96,6 ± 29,41 (7-846 e 93,3 ± 28,6 (0-846, respectivamente. Foi observada correlação positiva entre a abundância dos parasitas presentes nos olhos com relação ao comprimento padrão (rs = 0.5, p = 0.005, peso (rs = 0.649, p = 0.0001 e fator de condição (rs = 0.439, p = 0.0154. As altas taxas de parasitismo de metacercárias de S. musculosum em S. insculpta podem ser um indicativo de que esta espécie de peixe é altamente suscept

In-vivo monitoring of development of cholangiocarcinoma induced with C. sinensis and N-nitrosodimethylamine in Syrian golen hamsters using ultrasonography and magnetic resonance imaging: a preliminary study

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Woo, Hyunsik [SMG-SNU Boramae Medical Center, Department of Radiology, Seoul (Korea, Republic of); Han, Joon Koo; Kim, Jung Hoon [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Hong, Sung-Tae [Seoul National University, Department of Parasitology, College of Medicine, Seoul (Korea, Republic of); Uddin, M.H. [Seoul National University, Adult Stem Cell Research Center, Laboratory of Stem Cell and Tumor Biology, Department of Veterinary Public Health, College of Veterinary Medicine, Seoul (Korea, Republic of); Jang, Ja-June [Seoul National University, Department of Pathology, College of Medicine, Seoul (Korea, Republic of)

2017-04-15

The purpose of this study is to evaluate high-resolution ultrasound and magnetic resonance imaging (MRI) in monitoring of cholangiocarcinoma in the hamsters with C. sinensis infection and N-nitrosodimethylamine (NDMA). Twenty-four male Syrian golden hamsters of were divided into four groups composed of five hamsters as control, five hamsters receiving 30 metacercariae of C. sinensis per each hamster, five hamsters receiving NDMA in drinking water, and nine hamsters receiving both metacercariae and NDMA. Ultrasound was performed every other week from baseline to the 12th week of infection. MRI and histopathologic examination was done from the 4th week to 12th week. Cholangiocarcinomas appeared as early as the 6th week of infection. There were 12 cholangiocarcinomas, nine and ten of which were demonstrated by ultrasound and MRI, respectively. Ultrasound and MRI findings of cholangiocarcinomas in the hamsters were similar to those of the mass-forming intrahepatic cholangiocarcinomas in humans. Ultrasound and MRI also showed other findings of disease progression such as periductal increased echogenicity or signal intensity, ductal dilatation, complicated cysts, and sludges in the gallbladder. High-resolution ultrasound and MRI can monitor and detect the occurrence of cholangiocarcinoma in the hamsters non-invasively. (orig.)

Relationship between thermal loading and parasitism in the mosquitofish

International Nuclear Information System (INIS)

Aho, J.M.; Gibbons, J.W.; Esch, G.W.

1976-01-01

The relationship between thermal loading and parasitism was examined in 980 mosquitofish, Gambusia affinis, taken from areas of varying thermal conditions at the Savannah River Plant (SRP) near Aiken, S. C. Collections were made at 2-week intervals from May to August 1974 and again from January to February 1975. The metacercaria of two strigeid trematodes, Ornithodiplostomum ptychocheilus and Diplostomum scheuringi were the only species of parasites recovered. Ornithodiplostomum ptychocheilus was always found encysted in the brain and eyes, whereas D. scheuringi was restricted exclusively to the body cavity. The density of the body-cavity parasite was highest in fish from areas of ambient temperatures and declined in fish from areas with higher water temperatures. The density of the brain parasite, on the other hand, was higher in fish from areas directly receiving thermal effluent than in fish from ambient-temperature areas. The body-cavity parasite was absent from mosquitofish from Pond C, but the infection percentages were relatively consistent in fish from other areas in the Par Pond system regardless of water temperature. The brain metacercaria were recovered from 95 percent of fish from Pond C

In-vivo monitoring of development of cholangiocarcinoma induced with C. sinensis and N-nitrosodimethylamine in Syrian golen hamsters using ultrasonography and magnetic resonance imaging: a preliminary study

International Nuclear Information System (INIS)

Woo, Hyunsik; Han, Joon Koo; Kim, Jung Hoon; Hong, Sung-Tae; Uddin, M.H.; Jang, Ja-June

2017-01-01

The purpose of this study is to evaluate high-resolution ultrasound and magnetic resonance imaging (MRI) in monitoring of cholangiocarcinoma in the hamsters with C. sinensis infection and N-nitrosodimethylamine (NDMA). Twenty-four male Syrian golden hamsters of were divided into four groups composed of five hamsters as control, five hamsters receiving 30 metacercariae of C. sinensis per each hamster, five hamsters receiving NDMA in drinking water, and nine hamsters receiving both metacercariae and NDMA. Ultrasound was performed every other week from baseline to the 12th week of infection. MRI and histopathologic examination was done from the 4th week to 12th week. Cholangiocarcinomas appeared as early as the 6th week of infection. There were 12 cholangiocarcinomas, nine and ten of which were demonstrated by ultrasound and MRI, respectively. Ultrasound and MRI findings of cholangiocarcinomas in the hamsters were similar to those of the mass-forming intrahepatic cholangiocarcinomas in humans. Ultrasound and MRI also showed other findings of disease progression such as periductal increased echogenicity or signal intensity, ductal dilatation, complicated cysts, and sludges in the gallbladder. High-resolution ultrasound and MRI can monitor and detect the occurrence of cholangiocarcinoma in the hamsters non-invasively. (orig.)

Role of the lymnaeid snail Pseudosuccinea columella in the transmission of the liver fluke Fasciola hepatica in Egypt.

Science.gov (United States)

Dar, Y; Vignoles, P; Rondelaud, D; Dreyfuss, G

2015-11-01

Experimental infections of three Egyptian Pseudosuccinea columella populations with sympatric miracidia of Fasciola sp., coming from cattle- or sheep-collected eggs, were carried out to determine the capacity of this lymnaeid to support larval development of the parasite. Using microsatellite markers, the isolates of Egyptian miracidia were identified as Fasciola hepatica. Apart from being independent of snail origin, prevalences ranging from 60.4 to 75.5% in snails infected with five miracidia of F. hepatica were significantly higher than values of 30.4 to 42.2% in snails with bi-miracidial infections. The number of metacercariae ranged from 243 to 472 per cercarial-shedding snail and was independent of snail origin, parasite origin and miracidial dose used for infection. If P. columella was subjected to two successive bi-miracidial infections with F. hepatica, prevalence of infection was 63.3%, with a mean of 311 metacercariae per snail. These values were clearly greater than those already reported for Radix natalensis infected with the same parasite and the same protocol. Successful experimental infection of P. columella with F. hepatica suggests that this lymnaeid snail is an important intermediate host for the transmission of fascioliasis in Egypt.

Digenetic trematodes in South American sea lions from southern Brazilian waters.

Science.gov (United States)

Pereira, E M; Müller, G; Secchi, E; Pereira, J; Valente, A L S

2013-10-01

The aim of this work was to perform a systematic study to detect and quantify the digenetic trematode infections in South American sea lions from the southern Brazilian coast. Twenty-four South American sea lions, Otaria flavescens (Carnivora: Otaridae), were found dead along the coast of Rio Grande do Sul State, Brazil, between June 2010 and September of 2011. Two trematode species were found in the intestines of O. flavescens, i.e., Stephanoprora uruguayense (Digenea: Echinostomatidae) and Ascocotyle (Phagicola) longa (Digenea: Heterophyidae). Ascocotyle (P.) longa reached a prevalence of 33.3% and mean intensity of 248,500, whereas S. uruguayense showed a prevalence of 4.2% and mean intensity of 202. The 2 trematode species infecting sea lions were likely transmitted by feeding on mullets, Mugil platanus, that commonly harbor heterophyid metacercariae. The present work is the first report of digenetic trematodes infecting O. flavescens in Brazil. The high prevalence and mean intensity values of the 2 trematode species infecting sea lions in the present study suggest caution in human consumption of mullets and other fish, which can be infected with the metacercariae of these trematodes known to have zoonotic potential.

Identification, immunolocalization, and characterization analyses of an exopeptidase of papain superfamily, (cathepsin C) from Clonorchis sinensis.

Science.gov (United States)

Liang, Pei; He, Lei; Xu, Yanquan; Chen, Xueqing; Huang, Yan; Ren, Mengyu; Liang, Chi; Li, Xuerong; Xu, Jin; Lu, Gang; Yu, Xinbing

2014-10-01

Cathepsin C is an important exopeptidase of papain superfamily and plays a number of great important roles during the parasitic life cycle. The amino acid sequence of cathepsin C from Clonorchis sinensis (C. sinensis) showed 54, 53, and 49% identities to that of Schistosoma japonicum, Schistosoma mansoni, and Homo sapiens, respectively. Phylogenetic analysis utilizing the sequences of papain superfamily of C. sinensis demonstrated that cathepsin C and cathepsin Bs came from a common ancestry. Cathepsin C of C. sinensis (Cscathepsin C) was identified as an excretory/secretory product by Western blot analysis. The results of transcriptional level and translational level of Cscathepsin C at metacercaria stage were higher than that at adult worms. Immunolocalization analysis indicated that Cscathepsin C was specifically distributed in the suckers (oral sucker and ventral sucker), eggs, vitellarium, intestines, and testis of adult worms. In the metacercaria, it was mainly detected on the cyst wall and excretory bladder. Combining with the results mentioned above, it implies that Cscathepsin C may be an essential proteolytic enzyme for proteins digestion of hosts, nutrition assimilation, and immune invasion of C. sinensis. Furthermore, it may be a potential diagnostic antigen and drug target against C. sinensis infection.

Exploring the diversity of Diplostomum (Digenea: Diplostomidae) in fishes from the River Danube using mitochondrial DNA barcodes

Czech Academy of Sciences Publication Activity Database

Kudlai, Olena; Oros, M.; Kostadinova, Aneta; Georgieva, Simona

2017-01-01

Roč. 10, 2 December (2017), č. článku 592. ISSN 1756-3305 R&D Projects: GA ČR GA15-14198S; GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:60077344 Keywords : Diplostomum * Diplostomidae * Metacercariae * Freshwater fishes * Barcodes * cox1 * nad3 * River Danube * Europe Subject RIV: EG - Zoology OBOR OECD: Zoology Impact factor: 3.080, year: 2016

Identification and characterization of a member of Rab subfamily, Rab8, from Clonorchis sinensis.

Science.gov (United States)

Liang, Pei; He, Lei; Yu, Jinyun; Xie, Zhizhi; Chen, Xueqing; Mao, Qiang; Liang, Chi; Huang, Yan; Lu, Gang; Yu, Xinbing

2015-05-01

The Rabs act as a binary molecular switch that utilizes the conformational changes associated with the GTP/GDP cycle to elicit responses from target proteins. It regulates a broad spectrum of cellular processes including cell proliferation, cytoskeletal assembly, and intracellular membrane trafficking in eukaryotes. The Rab8 from Clonorchis sinensis (CsRab8) was composed of 199 amino acids. The deduced amino acid sequence shared above 50% identities with other species from trematode, tapeworm, mammal, insecta, nematode, and reptile, respectively. The homologous analysis of sequences showed the conservative domains: G1 box (GDSGVGKS), G2 box (T), G3 box (DTAG), G4 box (GNKCDL), and G5 box. In addition, the structure modeling had also shown other functional domains: GTP/Mg(2+) binding sites, switch I region, and switch II region. A phylogenic tree analysis indicated that the CsRab8 was clustered with the Rab from Schistosoma japonicum, and trematode and tapeworm came from the same branch, which was different from an evolutional branch built by other species, such as mammal animal, insecta, nematode, and reptile. The recombinant CsRab8 protein was expressed in Escherichia coli and the purified protein was a soluble molecule by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. CsRab8 was identified as a component of excretory/secretory products of C. sinensis by western blot analysis. The transcriptional level of CsRab8 at metacercaria stage was the highest at the four stages and higher by 56.49-folds than that at adult worm, 1.23-folds than that at excysted metacercaria, and 2.69-folds than that at egg stage. Immunohistochemical localization analysis showed that CsRab8 was specifically distributed in the tegument, vitellarium, eggs, and testicle of adult worms, and detected on the vitellarium and tegument of metacercaria. Combined with the results, CsRab8 is indispensable for survival and development of parasites, especially for regulating

[Investigation on sanitation of freshwater aquaculture environments and Clonorchis sinensis intermediate host infection in a city of Pearl River Delta region, China].

Science.gov (United States)

Man, Wang; Le, Luo; Xue-Qin, Chen; Lei, Li; Yue-Yi, Fang

2017-10-19

To understand the current status of the sanitation of freshwater aquaculture environments, and Clonorchis sinensis infection of freshwater fish in the aquaculture and market in a city of Pearl River Delta region, so as to provide the evidence for formulating the prevention and control strategy of clonorchiasis sinensis. In 2016, based on the distribution of freshwater aquaculture, 36 freshwater fish ponds among 14 towns were selected for sampling and investigation, and 10-20 pieces were collected from each pond. Besides, 3 aquatic product wholesale markets were included, among which 3-6 stalls were selected from each market, and 20-30 pieces were collected from each stall. The metacercaria in the fish was examined by the digestion method. In the 36 fish ponds, there were no toilets with the stool being drained into fish ponds directly, and there was only one pond with duck sheds with the stool being drained into fish ponds directly. Totally 437 pieces of freshwater fish from ponds were detected, with a metacercaria positive rate of 4.35% (19/437). The metacercaria positive fish were distributed in 50% (7/14) of towns and 25% (9/36) ponds. The positive rates of crucian carp, grass carp, dace, aristichthysnobilis, and tilapia were 13.95% (6/43), 4.76% (9/189), 4.44 (2/45), 1.55% (2/129), and 0 (0/31) respectively, with statistically significant difference ( χ 2 = 13.46, P = 0.01). Totally 307 pieces of freshwater fish were collected from the wholesale markets, with a total positive rate of 1.95% (6/307). The positive rate of grass carp and aristichthysnobilis were 3.20% (4/125) and 2.78% (2/72) respectively, and no positive samples were found in crucian carp, dace and tilapia, with no statistically significant difference among the different fish in the infection rate (Fisher exact P = 0.75). The sanitation of freshwater aquaculture environments in a city of Pearl River Delta region is relative good. However, there are different degrees of Clonorchis sinensis

Larval trematodes (Digenea) of the great pond snail, Lymnaea stagnalis (L.), (Gastropoda, Pulmonata) in Central Europe: a survey of species and key to their identification

Czech Academy of Sciences Publication Activity Database

Faltýnková, Anna; Našincová, Vanda; Kablásková, Lenka

2007-01-01

Roč. 14, č. 1 (2007), s. 39-51 ISSN 1252-607X R&D Projects: GA AV ČR IAA6022404; GA ČR GP524/07/P086; GA ČR GD524/03/H133; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trematoda * Mollusca * cercaria * metacercaria * Lymnaea stagnalis Subject RIV: EG - Zoology Impact factor: 0.713, year: 2007

Small-sized euryhaline fish as intermediate hosts of the digenetic trematode Cryptocotyle concavum

Science.gov (United States)

Zander, C. D.; Kollra, H.-G.; Antholz, B.; Meyer, W.; Westphal, D.

1984-03-01

Cercariae of the trematode Cryptocotyle concavum, which encyst in skin and/or kidney of sticklebacks and gobies, were studied in the Schlei Fjord (western Baltic Sea). Mean incidence of dermal cysts was 48 % in Gasterosteus aculeatus and 37 % in Pungitius pungitius. No cysts were found in the kidneys of sticklebacks. While 97 % of Pomatoschistus microps had encysted metacercariae in the kidneys, only 2 % had cysts in the skin. Pomatoschistus minutus, however, showed hardly any cyst infestation of either skin or kidney. In P. microps the intensity of infestation by metacercariae was frequently more than 50 cysts; in contrast, sticklebacks rarely exhibited more than 5 dermal cysts. Infested fish were larger than 10 mm in total length, the incidence rate increasing with growth. Parasitic infestation depends on ambient salinity: C. concavum was not found at salinities below 4 ‰. In contrast to the high incidence in fish, the first hosts — the snails Hydrobia stagnalis and H. neglecta — showed remarkably low infection rates (3 to 5 %). The findings reported are related to the distribution of C. concavum, the mode of life of infested fish, the feeding habits of the final hosts and the infestation of P. microps by other parasites. Evidently, P. microps represents an optimal second host for C. concavum.

Prevalence, geographic distribution, and fitness effects of Microphallus turgidus (trematoda: Microphallidae) in grass shrimp (Palaemonetes spp.) from coastal Georgia.

Science.gov (United States)

Pung, Oscar J; Khan, Ritindra N; Vives, Stephen P; Walker, Casey B

2002-02-01

Grass shrimp, Palaemonetes pugio Holthuis and P. vilgaris (Say), were collected at 11 localities along the coast of Georgia and surveyed for digenetic trematode metacercariae. The effect of trematode infection on grass shrimp fitness was also examined. Microphallus turgidas (Leigh) was the only trematode observed. The prevalence of metacercarial cysts of this parasite in P. pugio (75%) was higher than in P. vulgaris (24%), as were the mean intensity, abundance, and population density (no. cysts/cm host body length) of the parasite. Infected shrimp were found at every collection locality and parasite prevalence and density were greatest in P. pugio from higher salinity localities (> or = 20 parts per thousand [ppt]). There was no relation between host body size and parasite density in P. vulgaris, and parasite density increased with host body size in P. pugio, suggesting that the parasite does not affect host survival. There was no relation between parasite density and shrimp egg mass, but nonovigerous female P. pugio were more heavily infected than ovigerous ones. In addition, 1 metacercaria of M. turgidus in each of 2 specimens of P. vulgaris was parasitized by the haplosporidian Urosporidium crescens De Turk. This represents a new shrimp host record for this hyperparasite.

Association between Opisthorchis viverrini and Leptospira spp. infection in endemic Northeast Thailand.

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Van, Chinh Dang; Doungchawee, Galayanee; Suttiprapa, Sutas; Arimatsu, Yuji; Kaewkes, Sasithorn; Sripa, Banchob

2017-08-01

Opisthorchiasis caused by Opisthorchis viverrini is an important foodborne trematodiasis in Thailand, Laos and Cambodia. Interestingly, the opisthorchiasis endemic region overlaps with an area of leptospirosis emergence. Here we report an association between opisthorchiasis and leptospirosis in Thailand. Of 280 sera collected from villagers living around the Lawa wetland complex in Khon Kaen province, 199 (71%) were seropositive for leptospirosis by immunochromatography. Individuals with O. viverrini infection had a significantly higher rate of leptospirosis than those without (P=0.001). Significant higher leptospirosis prevalence was found in males than females (P=0.002). However, females but not males with O. viverrini infection showed a significantly higher seroprevalence of leptospirosis. Twenty-one of 35 environmental samples from the lake (water, mud and fish skin mucus) were positive for Leptospira spp. DNA sequencing, sequence alignment, and phylogenetic analysis of some positive nested PCR products revealed both pathogenic and intermediate pathogenic strains of Leptospira in the samples. Strikingly, O. viverrini metacercariae from the fish were positive for L. interrogans. These results suggest a close association between opisthorchiasis and leptospirosis. Contact with water, mud or eating raw fish harboring liver fluke metacercariae may be risk factors for Leptospira infection. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Parasites of ornamental fish commercialized in Macapá, Amapá State (Brazil

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Érico de Melo Hoshino

2018-02-01

Full Text Available Abstract This study investigated the parasites fauna of four freshwater ornamental fish species in aquarium shops of Macapá, Amapá State, in addition to survey the commercialized fish species and sanitary conditions of aquarium shops. Different native and non-native ornamental fish species were found in aquarium shops, mainly Poecilidae. We examined 30 specimens of Xiphophorus maculatus, 30 Danio rerio, 30 Paracheirodon axelrodi, and 30 Corydoras ephippifer for parasites. Of the 120 fish examined, 22.5% were parasitized by one or more species and a total of 438 parasites were collected and identified. Parasites such as: Ichthyophthirius multifiliis, Monogenea, undermined Digenea metacercariae, Acanthostomum sp. metacercariae, Camallanus spp., Bothriocephalus acheilognathi and Echinorhynchus sp. infected the hosts examined. Endoparasites in the larval stage showed the greatest diversity and Camallanus spp. was found in all hosts species examined. Paracheirodon axelrodi (43.3% was the most parasitized host, while C. ephippifer (6.7% was the least parasitized. Despite the low ectoparasites level, six species of endoparasites was observed, demonstrating that prophylactic and quarantine procedures were not fully adequate. Therefore, failures in prophylactic procedures on any link in the production industry of ornamental fish may cause parasite transmission to ornamental fish captured in different environments and localities.

Aspects of the Maintenance of the Life Cycle of Fasciola hepatica in Lymnaea columella in Minas Gerais, Brazil

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Souza Cecília Pereira de

2002-01-01

Full Text Available Fascioliasis is a parasitic disease of domestic ruminants that occurs worldwide. The lymnaeid intermediate hosts of Fasciola hepatica include Lymnaea columella, which is widely distributed in Brazil. A colony of L. columella from Belo Horizonte, MG, was reared in our laboratory to be used in studies of the F. hepatica life cycle, the intermediate host-parasite relationship and development of an anti-helminthic vaccine. In the first experiment 1,180 snails were exposed to miracidia of F. hepatica eggs removed from the biliary tracts of cattle from the State of Rio Grande do Sul. In the second and third experiments the snails were exposed to miracidia that had emerged from F. hepatica eggs from Uruguay, maintained in rabbits. The rates of infection in the first, second and third experiments were 0, 42.1 and 0% respectively. Over 15,806 metacercariae were obtained and stored at 4ºC. Four rabbits weighing 1.5 kg each were infected with 32-44 metacercariae and two with 200. Three rabbits begin to eliminate eggs of the parasite in the feces from 84 days after infection onwards. The biological cycle of F. hepatica in L. columella and the rabbit was completed within 124 days.

Host specificity of Pisidium coreanum (Bivalvia: Sphaeriidae) to larval infection with a human intestinal fluke Echinostoma cinetorchis (Trematoda: Echinostomatidae) in Korea.

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Park, Y K; Soh, C T; Park, G M; Hwang, M K; Chung, P R

2006-10-01

The fingernail clam, Pisidium coreanum, has been traditionally consumed raw as a so-called drug therapy by patients with bone fractures in Korea. The present study was designed to determine the possible occurrence and, if present, the prevalence of Echinostoma cinetorchis in P. coreanum collected at a local site, and to determine the susceptibility of the clams in the laboratory to infection with miracidia and cercariae of E. cinetorchis. No cercariae or metacercariae of E. cinetorchis were observed in field-collected P. coreanum clams. In susceptibility experiments with laboratory-reared clams, individuals exposed to miracidia of E. cinetorchis did not release cercariae by 20 days after exposure; necropsy of exposed clams failed to show development of any sporocysts or rediae. To confirm the possibility of these clams serving as an experimental second intermediate host of E. cinetorchis, 20 of them were exposed to E. cinetorchis cercariae from experimentally infected Segmentina hemisphaerula that had been previously exposed to miracidia of E. cinetorchis; all exposed clams became infected. Metacercariae from clams at 14 days postinfection were fed to rats, and adult worms were recovered from the ileocecal regions. This is the first report of P. coreanum serving as second intermediate host of E. cinetorchis.

Identification and molecular characterization of Parkin in Clonorchis sinensis.

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Bai, Xuelian; Kim, Tae Im; Lee, Ji-Yun; Dai, Fuhong; Hong, Sung-Jong

2015-02-01

Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn(2+) were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

Hypolobocera guayaquilensis (Decapoda: Pseudothelphusidae): A New Crab Intermediate Host of Paragonimus mexicanus in Manabí Province, Ecuador

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Calvopina, Manuel; Romero-Alvarez, Daniel; Rendon, Melina; Takagi, Hidekazu; Sugiyama, Hiromu

2018-01-01

To determine that Paragonimus sp. is actively transmitted in a tropical area of the Pacific region of Ecuador where human cases of pulmonary paragonimiasis have recently been documented, a total of 75 freshwater crabs were collected from 2 different streams in the Pedernales area of Manabí Province, Ecuador. All collected crabs were identified as Hypolobocera guayaquilensis based on morphological characteristics of the male gonopods. The hepatopancreas of each crab was examined by compressing it between 2 glass plates followed by observation under a stereomicroscope. Excysted Paragonimus metacercariae were detected in 39 (52.0%) crabs and their densities varied from 1 to 32 per infected crab. There was a positive relationship between crab size and metacercarial density. Sequences of the second internal transcribed spacer region of the ribosomal RNA gene of the Paragonimus metacercariae obtained in this study were identical to those of Paragonimus mexicanus deposited in the DDBJ/EMBL/GenBank database. Thus, the present study is the first to confirm that the crab species H. guayaquilensis is the second intermediate host of P. mexicanus in Manabí Province, Ecuador. Because this crab might be the possible source of human infections in this area, residents should pay attention to improper crab-eating habits related with a neglected parasitic disease, i.e., paragonimiasis. PMID:29742874

Defeating diplostomoid dangers in USA catfish aquaculture.

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Overstreet, Robin M; Curran, Stephen S

2004-06-01

Diplostomoid digenean metacercariae have caused widescale mortalities of channel catfish, Ictalurus punctatus (Rafinesque), at aquaculture farms in Louisiana, Mississippi and Arkansas, USA. Originally, based on a tentative diagnosis, the industry considered the primary harmful agent to be an introduced species from Europe, Bolbophorus confusus (Krause, 1914), frequently reported from the American white pelican, Pelecanus erythrorhynchos Gmelin. Our group has now shown, using ITS 1-2 plus three more-conservative gene fragments, that two sympatric species of Bolbophorus exist in the American white pelican. One, B. damnificus Overstreet et Curran, 2002, infects the musculature of catfish, and the other, probably not B. confusus, does not infect catfish. However, at least four other pathogenic diplostomoids and a clinostomoid infect the catfish, and they use at least four different snail hosts, including the planorbids Planorbella trivolvis (Say) and Gyraulus parvus (Say), the physid Physella gyrina (Say) and a lymnaeid. Two metacercariae, B. damnificus and Bursacetabulus pelecanus Dronen, Tehrany et Wardle, 1999, infect the catfish and mature in the pelican; two others, Austrodiplostomum compactum (Lutz, 1928) and Hysteromorpha cf. triloba (Rudolphi, 1819), mature in cormorants; one, Diplostomum sp., matures in seagulls and at least one, Clinostomum marginatum (Rudolphi, 1819), matures in herons, egrets and other wading birds. Consequently, management of catfish ponds relative to digenean infections requires considerable biological information on the fish, bird, and snail hosts as well as the parasites.

Biology of Crassicutis cichlasomae, a parasite of cichlid fishes in Mexico and Central America.

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Scholz, T; Pech-Ek, M C; Rodriguez-Canul, R

1995-03-01

Field study on the biology of Crassicutis cichlasomae Manter, 1936 (Digenea: Homalometridae) was carried out in a small swamp in a limestone factory near Mérida, Yucatán, Mexico. Aquatic snails, Littorina (Littoridinopis) angulifera, harbouring C. cichlasomae rediae, cercariae and metacercariae, served both as the first and second intermediate hosts. Feeding experiments confirmed the conspecificity of metacercariae from naturally infected snails with adults from naturally infected fish. Gravid C. cichlasomae worms were obtained from experimentally infected fish 19 days post exposure at 22-24 degrees C. Examination of fish from the swamp in Mitza and other localities in the Yucatan Peninsula showed that the cichlids Cichlasoma urophthalmus and C. meeki were definitive hosts of C. cichlasomae. There was no pronounced preference of C. cichlasomae adults for the site of their location in the intestine of the definitive host; a slightly higher proportion (41%) of worms was only found in the anterior third of the gut. The time of miracidium development varied from 18.5 to 27.5 days; different temperature (20.1-35.7 degrees C) or light/darkness regimes influenced only slightly the rate of embryonic development, with shorter development times at higher temperature (34.8-35.7 degrees C) and constant darkness and/or light. With the exception of the sporocyst, all developmental stages are described and figured.

Comparative strategies and success of sympatric and allopatric Fasciola hepatica infecting Galba truncatula of different susceptibilities.

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Sanabria, R; Mouzet, R; Courtioux, B; Vignoles, P; Rondelaud, D; Dreyfuss, G; Cabaret, J; Romero, J

2013-06-01

Allopatric infections of French Galba truncatula with an Argentinean isolate of Fasciola hepatica were carried out to determine the infectivity of foreign miracidia in three snail populations differing by their susceptibility to French miracidia (two highly and one poorly susceptible populations). Sympatric infections of G. truncatula with French miracidia were used as controls. Compared to sympatric infections of G. truncatula, snail survival at day 30 post-infection in allopatric groups was significantly lower in a highly susceptible population and significantly greater in the other two. Prevalence in snails infected with the allopatric isolate was significantly lower (16.4-34.5 % instead of 58.6-72.1 %), whereas their patent period was significantly longer (a mean of 69.9-85.9 days instead of 6.4-20.7 days). The mean number of metacercariae was also higher in allopatric groups (236.5-897.3 per cercariae-shedding snail instead of 70.7-222.1). Owing to longer patent periods, the Argentinean isolate of F. hepatica was less pathogenic for these snails. The lower prevalence of infection, the longer patent period and the higher number of metacercariae noted in allopatric groups might be the consequence of an adaptive mechanism used by this digenean introduced to the New World to infect new populations of unusual intermediate hosts.

Efficacy of praziquantel against Clonorchis sinensis infection in dogs and cats.

OpenAIRE

斎藤, 哲郎; 森重, 和久; 川崎, 伸二; 頓宮, 廉正

1993-01-01

The efficacy of praziquantel was tested using 11 dogs and 3 cats infected with Clonorchis sinensis. Each experimental animal was infected with 50 metacercariae of Clonorchis sinensis respectively. The efficacy of the drug was evaluated by considerable reductions of EPG. The worms were eliminated from 91% of dogs and 100% of cats with hypodermic injection of total 75mg/kg praziquantel. This drug was effective against Clonorchis sinensis infection in dogs and cats as in the case of other tremat...

Description and proposed life cycle of Maritrema novaezealandensis n. sp. (Microphallidae) parasitic in red-billed gulls, Larus novaehollandiae scopulinus, from Otago Harbor, South Island, New Zealand

DEFF Research Database (Denmark)

Martorelli, Sergio R; Fredensborg, Brian Lund; Mouritsen, Kim Nørgaard

2004-01-01

Maritrema novaezealandensis n. sp. is described from Otago Harbor, South Island, New Zealand, on the basis of adult specimens collected from the Red-billed gull, Larus novaehollandiae scopulinus, and excysted metacercariae obtained from crabs. It belongs to the "eroliae group" and differs from...... snail, Zeacumantus subcarinatus, in which the cercarial stage is produced in sporocysts located within the gonad of the snail. At least 3 crab species (Hemigrapsus crenulatus, Macrophtalmus hirtipes, and Halicarcinus whitei) and several species of amphipods act as second intermediate hosts...

Gamma ionization of Phagicola longa (Trematoda:Heterophyidae) in Mugilidae (pisces) in São Paulo, Brazil

Science.gov (United States)

Antunes, S. A.; Wiendl, F. M.; Almeida Dias, E. R.; Arthur, V.; Daniotti, C.

1993-07-01

The mullet (Mugilidae) is a fish caught in large quantities in brackish and marine waters of the southern Brazilian coast, which in consequence of its raw consumption as "sashimi", a typical Japanese dish, has led to ten cases of human infection by Phagicola longa in São Paulo, Brazil. The mullet acts as a second intermediate host for the parasite, as in human heterophysiasis in the Near and Far East. In order to control these infections under commercial storage conditions and in raw consumption, the radiolysis of P. longa was studied in three mullet species-the silver mullet ( Mugil curema), the grey mullet ( M. platanus) and the "paratipema" ( Mugil sp.) - subjected to ionization ranging from 1.0 to 10.0 kGy (2.21 kGy/h). It was observed that 1.0 and 2.0 kGy caused a motility decrease in silver mullet parasites from 100% to 15% and 17%, and that doses of 4.0 and 10.0 kGy caused metacercaria inviability. The parasite motility decreased in the grey mullet treated with doses of 2.0, 2.5, 3.0 and 3.5 kGy, from 56% to 31%, 9%, 18% and 5%, respectively, 4.0 kGy tending to be the control dose for P. longa. This dose also controls other metacercaria found in the "parati-pema", without changing the odor, color or appearance of the treated mullet.

Gamma ionization of Phagicola longa (Trematoda:Heterophyidae) in Mugilidae (pisces) in Sao Paulo, Brazil

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Antunes, S.A.; Wiendl, F.M.; Almeida Dias, E.R.; Arthur, V.; Daniotti, C. (Sao Paulo Univ., SP (Brazil))

The mullet (Mugilidae) is a fish caught in large quantities in brackish and marine waters of the southern Brazilian coast, which in consequence of its raw consumption as ''sashimi'', a typical Japanese dish, has led to ten cases of human infection by Phagicola longa in Sao Paulo, Brazil. The mullet acts as a second intermediate host for the parasite, as in human heterophyiasis in the Near and Far East. In order to control these infections under commercial storage conditions and in raw consumption, the radiolysis of P. longa was studied in three mullet species - the silver mullet (Mugil curema), the grey mullet (M. platanus) and the ''paratipema'' (Mugil sp.) -subjected to ionization ranging from 1.0 to 10.0 kGy (2.21 kGy/h). It was observed that 1.0 and 2.0 kGy caused a motility decrease in silver mullet parasites from 100% to 15% and 17%, and that doses of 4.0 and 10.0 kGy caused metacercaria inviability. The parasite motility decreased in the grey mullet treated with doses of 2.0, 2.5, 3.0 and 3.5 kGy, from 56% to 31%, 9%, 18% and 5%, respectively, 4.0 kGy tending to be the control dose for P. Longa. This dose also controls other metacercaria found in the ''parati-pema'', without changing the odor, color or appearance of the treated mullet. (author).

Gamma ionization of Phagicola longa (Trematoda:Heterophyidae) in Mugilidae (pisces) in Sao Paulo, Brazil

International Nuclear Information System (INIS)

Antunes, S.A.; Wiendl, F.M.; Almeida Dias, E.R.; Arthur, V.; Daniotti, C.

1993-01-01

The mullet (Mugilidae) is a fish caught in large quantities in brackish and marine waters of the southern Brazilian coast, which in consequence of its raw consumption as ''sashimi'', a typical Japanese dish, has led to ten cases of human infection by Phagicola longa in Sao Paulo, Brazil. The mullet acts as a second intermediate host for the parasite, as in human heterophyiasis in the Near and Far East. In order to control these infections under commercial storage conditions and in raw consumption, the radiolysis of P. longa was studied in three mullet species - the silver mullet (Mugil curema), the grey mullet (M. platanus) and the ''paratipema'' (Mugil sp.) -subjected to ionization ranging from 1.0 to 10.0 kGy (2.21 kGy/h). It was observed that 1.0 and 2.0 kGy caused a motility decrease in silver mullet parasites from 100% to 15% and 17%, and that doses of 4.0 and 10.0 kGy caused metacercaria inviability. The parasite motility decreased in the grey mullet treated with doses of 2.0, 2.5, 3.0 and 3.5 kGy, from 56% to 31%, 9%, 18% and 5%, respectively, 4.0 kGy tending to be the control dose for P. Longa. This dose also controls other metacercaria found in the ''parati-pema'', without changing the odor, color or appearance of the treated mullet. (author)

A case of ectopic intraabdominal fascioliasis presented with acute abdomen.

Science.gov (United States)

Tanir, Gönül; Karaman, Ayşe; Tüfekçı, Sehra Birgül; Erdoğan, Duygu; Tuygun, Nilden; Ozkan, Ayşegül Taylan

2011-06-01

Human fascioliasis with Fasciola species occurs worldwide and is most common among rural people who tend sheep and eat uncooked water vegetables, particularly watercress. The natural history of the acute phase begins with ingestion of metacercariae encysted on various kinds of aquatic vegetation such as watercress. Fascioliasis primarily involves the liver, bile ducts, gallbladder, and occasionally ectopic sites. We describe herein a case of ectopic fascioliasis. This uncommon form of disease was peritonitis; both visceral and parietal peritoneal layers were affected with the formation of multiple nodules and ascites.

Determination of the Minimum Effective Dosages of Praziquantel, Albendazole, and Mebendazole Against Clonorchis Sinensis Infection in Rats

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Ping-Chin Fan

2005-10-01

Full Text Available In order to determine the minimum effective dosages of praziquantel, albendazole, and mebendazole against Clonorchis sinensis infection in Sprague-Dawley rats, each rat was infected with 30 metacercariae and treated with one of three drugs. The rats were killed and examined 25 days after praziquantel treatment or 11 days after albendazole or mebendazole treatment. The minimum effective dosages were a single dose of praziquantel 375 mg/kg, albendazole 150 mg/kg, and mebendazole 150 mg/kg. Trials are required to determine whether these dosages are useful in the treatment of human clonorchiasis.

Fasciola hepatica: comparative metacercarial productions in experimentally-infected Galba truncatula and Pseudosuccinea columella

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Vignoles Philippe

2015-01-01

Full Text Available As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347–453 per cercariae-shedding snail versus 163–275, respectively. Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.

Cytosolic superoxide dismutase can provide protection against Fasciola gigantica.

Science.gov (United States)

Jaikua, Wipaphorn; Kueakhai, Pornanan; Chaithirayanon, Kulathida; Tanomrat, Rataya; Wongwairot, Sirima; Riengrojpitak, Suda; Sobhon, Prasert; Changklungmoa, Narin

2016-10-01

Superoxide dismutases (SOD), antioxidant metallo-enzymes, are a part of the first line of defense in the trematode parasites which act as the chief scavengers for reactive oxygen species (ROS). A recombinant Fasciola gigantica cytosolic SOD (FgSOD) was expressed in Escherichia coli BL21 (DE3) and used for immunizing rabbits to obtain polyclonal antibodies (anti-rFgSOD). This rabbit anti-rFgSOD reacted with the native FgSOD at a molecular weight of 17.5kDa. The FgSOD protein was expressed at high level in parenchyma, caecal epithelium and egg of the parasite. The rFgSOD reacted with antisera from rabbits infected with F. gigantica metacercariae collected at 2, 5, and 7 weeks after infection, and reacted with sera of infected mice. Anti-rFgSOD exhibited cross reactivity with the other parasites' antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. A vaccination was performed in imprinting control region (ICR) mice by subcutaneous injection with 50μg of rFgSOD combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 15 metacercariae by oral route. IgG1 and IgG2a in the immune sera were determined to indicate Th2 and Th1 immune responses. It was found that the parasite burden was reduced by 45%, and both IgG1 and IgG2a levels showed correlation with the numbers of worm recoveries. Copyright © 2016 Elsevier B.V. All rights reserved.

Molecular Cloning and Characterization of Growth Factor Receptor Bound-Protein in Clonorchis sinensis

Science.gov (United States)

Bai, Xuelian; Lee, Ji-Yun; Kim, Tae Im; Dai, Fuhong; Lee, Tae-Jin; Hong, Sung-Jong

2014-01-01

Background Clonorchis sinensis causes clonorchiasis, a potentially serious disease. Growth factor receptor-bound protein 2 (Grb2) is a cytosolic protein conserved among animals and plays roles in cellular functions such as meiosis, organogenesis and energy metabolism. In the present study, we report first molecular characters of growth factor receptor bound-protein (CsGrb2) from C. sinensis as counter part of Grb2 from animals and its possible functions in development and organogenesis of C. sinensis. Methodology/Principal Findings A CsGrb2 cDNA clone retrieved from the C. sinensis transcriptome encoded a polypeptide with a SH3-SH2-SH3 structure. Recombinant CsGrb2 was bacterially produced and purified to homogeneity. Native CsGrb2 with estimated molecular weight was identified from C. sinensis adult extract by western blotting using a mouse immune serum to recombinant CsGrb2. CsGrb2 transcripts was more abundant in the metacercariae than in the adults. Immunohistochemical staining showed that CsGrb2 was localized to the suckers, mesenchymal tissues, sperms in seminal receptacle and ovary in the adults, and abundantly expressed in most organs of the metacercariae. Recombinant CsGrb2 was evaluated to be little useful as a serodiagnostic reagent for C. sinesis human infections. Conclusion Grb2 protein found in C. sinensis was conserved among animals and suggested to play a role in the organogenesis, energy metabolism and mitotic spermatogenesis of C. sinensis. These findings from C. sinensis provide wider understanding on diverse function of Grb2 in lower animals such as platyhelminths. PMID:24454892

Multidrug resistance-associated protein 4 is a bile transporter of Clonorchis sinensis simulated by in silico docking.

Science.gov (United States)

Dai, Fuhong; Yoo, Won Gi; Lee, Ji-Yun; Lu, Yanyan; Pak, Jhang Ho; Sohn, Woon-Mok; Hong, Sung-Jong

2017-11-21

Multidrug resistance-associated protein 4 (MRP4) is a member of the C subfamily of the ABC family of ATP-binding cassette (ABC) transporters. MRP4 regulates ATP-dependent efflux of various organic anionic substrates and bile acids out of cells. Since Clonorchis sinensis lives in host's bile duct, accumulation of bile juice can be toxic to the worm's tissues and cells. Therefore, C. sinensis needs bile transporters to reduce accumulation of bile acids within its body. We cloned MRP4 (CsMRP4) from C. sinensis and obtained a cDNA encoding an open reading frame of 1469 amino acids. Phylogenetic analysis revealed that CsMRP4 belonged to the MRP/SUR/CFTR subfamily. A tertiary structure of CsMRP4 was generated by homology modeling based on multiple structures of MRP1 and P-glycoprotein. CsMRP4 had two membrane-spanning domains (MSD1 & 2) and two nucleotide-binding domains (NBD1 & 2) as common structural folds. Docking simulation with nine bile acids showed that CsMRP4 transports bile acids through the inner cavity. Moreover, it was found that CsMRP4 mRNA was more abundant in the metacercariae than in the adults. Mouse immune serum, generated against the CsMRP4-NBD1 (24.9 kDa) fragment, localized CsMRP4 mainly in mesenchymal tissues and oral and ventral suckers of the metacercariae and the adults. Our findings shed new light on MRPs and their homologs and provide a platform for further structural and functional investigations on the bile transporters and parasites' survival.

Molecular cloning and characterization of growth factor receptor bound-protein in Clonorchis sinensis.

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Xuelian Bai

Full Text Available BACKGROUND: Clonorchis sinensis causes clonorchiasis, a potentially serious disease. Growth factor receptor-bound protein 2 (Grb2 is a cytosolic protein conserved among animals and plays roles in cellular functions such as meiosis, organogenesis and energy metabolism. In the present study, we report first molecular characters of growth factor receptor bound-protein (CsGrb2 from C. sinensis as counter part of Grb2 from animals and its possible functions in development and organogenesis of C. sinensis. METHODOLOGY/PRINCIPAL FINDINGS: A CsGrb2 cDNA clone retrieved from the C. sinensis transcriptome encoded a polypeptide with a SH3-SH2-SH3 structure. Recombinant CsGrb2 was bacterially produced and purified to homogeneity. Native CsGrb2 with estimated molecular weight was identified from C. sinensis adult extract by western blotting using a mouse immune serum to recombinant CsGrb2. CsGrb2 transcripts was more abundant in the metacercariae than in the adults. Immunohistochemical staining showed that CsGrb2 was localized to the suckers, mesenchymal tissues, sperms in seminal receptacle and ovary in the adults, and abundantly expressed in most organs of the metacercariae. Recombinant CsGrb2 was evaluated to be little useful as a serodiagnostic reagent for C. sinesis human infections. CONCLUSION: Grb2 protein found in C. sinensis was conserved among animals and suggested to play a role in the organogenesis, energy metabolism and mitotic spermatogenesis of C. sinensis. These findings from C. sinensis provide wider understanding on diverse function of Grb2 in lower animals such as platyhelminths.

Molecular identification, immunolocalization, and characterization of Clonorchis sinensis triosephosphate isomerase.

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Zhou, Juanjuan; Liao, Hua; Li, Shan; Zhou, Chenhui; Huang, Yan; Li, Xuerong; Liang, Chi; Yu, Xinbing

2015-08-01

Clonorchis sinensis triosephosphate isomerase (CsTIM) is a key regulatory enzyme of glycolysis and gluconeogenesis, which catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. In this study, the biochemical characterizations of CsTIM have been examined. A full-length complementary DNA (cDNA; Cs105350) sequence encoding CsTIM was obtained from our C. sinensis cDNA library. The open reading frame of CsTIM contains 759 bp which encodes 252 amino acids. The amino acid sequence of CsTIM shares 60-65% identity with other species. Western blot analysis displayed that recombinant CsTIM (rCsTIM) can be probed by anti-rCsTIM rat serum and anti-C. sinensis excretory/secretory products (anti-CsESPs) rat serum. Quantitative reverse transcription (RT)-PCR and western blotting analysis revealed that CsTIM messenger RNA (mRNA) and protein were differentially expressed in development cycle stages of the parasite, including adult worm, metacercaria, excysted metacercaria, and egg. In addition, immunolocalization assay showed that CsTIM was located in the seminal vesicle, eggs, and testicle. Moreover, rCsTIM exhibited active enzyme activity in catalytic reactions. The Michaelis constant (K m) of rCsTIM was 0.33 mM, when using glyceraldehyde 3-phosphate as the substrate. The optimal temperature and pH of CsTIM were 37 °C and 7.5-9.5, respectively. Collectively, these results suggest that CsTIM is an important protein involved in glycometabolism, and CsTIM possibly take part in many biological functions in the growth and development of C. sinensis.

The effect of x-irradiation on the development of Fasciola gigantica (cobbold, 1885) in goats

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Ravi Chandra

1976-01-01

The effect of varying doses (1 kr, 3 kr, 5 kr) of x-irradiation on the development of Fasciola gigantica, the common liver fluke of livestock in India has been investigated in goat kids with a view to develop a radiation attenuated vaccine against the pathogen. The growth and development of the flukes recovered from animals infected with irradiated metacercarie is found to be retarded. Metacecariae exposed to 3 kr can develop further, but number of flukes recovered from animals infected with metacecariae exposed to 5 kr is considerably less than the one from animals infected with non-irradiated metacercariae. (M.G.B.)

Survey of parasites in threatened stocks of coho salmon (Oncorhynchus kisutch) in Oregon by examination of wet tissues and histology.

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Ferguson, Jayde A; St-Hilaire, Sophie; Peterson, Tracy S; Rodnick, Kenneth J; Kent, Michael L

2011-12-01

We are conducting studies on the impacts of parasites on Oregon coastal coho salmon (Oncorhynchus kistuch). An essential first step is documenting the geographic distribution of infections, which may be accomplished by using different methods for parasite detection. Thus, the objectives of the current study were to (1) identify parasite species infecting these stocks of coho salmon and document their prevalence, density, and geographic distribution; (2) assess the pathology of these infections; and (3) for the first time, determine the sensitivity and specificity of histology for detecting parasites compared with examining wet preparations for muscle and gill infections. We examined 576 fry, parr, and smolt coho salmon in total by histology. The muscle and gills of 219 of these fish also were examined by wet preparation. Fish were collected from 10 different locations in 2006-2007. We identified 21 different species of parasites in these fish. Some parasites, such as Nanophyetus salmincola and Myxobolus insidiosus, were common across all fish life stages from most basins. Other parasites, such as Apophallus sp., were more common in underyearling fish than smolts and had a more restricted geographic distribution. Additional parasites commonly observed were as follows: Sanguinicola sp., Trichodina truttae , Epistylis sp., Capriniana piscium, and unidentified metacercariae in gills; Myxobolus sp. in brain; Myxidium salvelini and Chloromyxum majori in kidney; Pseudocapillaria salvelini and adult digenean spp. in the intestine. Only a few parasites, such as the unidentified gill metacercariae, elicted overt pathologic changes. Histology had generally poor sensitivity for detecting parasites; however, it had relatively good specificity. We recommend using both methods for studies or monitoring programs requiring a comprehensive assessment of parasite identification, enumeration, and parasite-related pathology.

Biochemical and immunological characterization of annexin B30 from Clonorchis sinensis excretory/secretory products.

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He, Lei; Ren, Mengyu; Chen, Xueqing; Wang, Xiaoyun; Li, Shan; Lin, Jinsi; Liang, Chi; Liang, Pei; Hu, Yue; Lei, Huali; Bian, Meng; Huang, Yan; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing

2014-07-01

Clonorchis sinensis has been classified as group I biological carcinogen for cholangiocarcinoma by the World Health Organization. Biological studies on excretory/secretory products (ESPs) enabled us to understand the pathogenesis mechanism of C. sinensis and develop new strategies for the prevention of clonorchiasis. In this study, sequence analysis showed that annexin B30 from C. sinensis (CsANXB30) is composed of four annexin repeats which were characterized by type II and III Ca(2+)-binding sites or KGD motif with the capability of Ca(2+)-binding. In addition, immunoblot assay revealed that recombinant CsANXB30 (rCsANXB30) could be recognized by the sera from rats infected with C. sinensis and the sera from rats immunized by CsESPs. Real-time PCR showed that its transcriptional level was the highest at the stage of metacercaria. Immunofluorescence assay was employed to confirm that CsANXB30 was distributed in the tegument, intestine, and egg of adult worms, as well as the tegument and vitellarium of metacercaria. rCsANXB30 was able to bind phospholipid in a Ca(2+)-dependent manner and human plasminogen in a dose-dependent manner. Moreover, cytokine and antibody measurements indicated that rats subcutaneously immunized with rCsANXB30 developed a strong IL-10 production in spleen cells and a high level of IgG1 isotype, indicating that rCsANXB30 could trigger specific humoral and cellular immune response in rats. The present results implied that CsANXB30 might be involved in a host-parasite interaction and affected the immune response of the host during C. sinensis infection.

Oral delivery of Bacillus subtilis spores expressing cysteine protease of Clonorchis sinensis to grass carp (Ctenopharyngodon idellus): Induces immune responses and has no damage on liver and intestine function.

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Tang, Zeli; Sun, Hengchang; Chen, TingJin; Lin, Zhipeng; Jiang, Hongye; Zhou, Xinyi; Shi, Cunbin; Pan, Houjun; Chang, Ouqin; Ren, Pengli; Yu, Jinyun; Li, Xuerong; Xu, Jin; Huang, Yan; Yu, Xinbing

2017-05-01

Clonorchis sinensis (C. sinensis) is a fish-borne trematode. Human can be infected by ingestion of C. sinensis metacercariae parasitized in grass carp (Ctenopharyngodon idella). For induction of effective oral immune responses, spores of Bacillus subtilis (B. subtilis) WB600 were utilized as vehicle to delivery CsCP (cysteine protease of C. sinensis) cooperated with CotC (B.s-CotC-CP), one of coat proteins, to the gastrointestinal tract. After routine culture of 8-12 h in LB medium, B. subtilis containing CotC-CsCP was transferred into the sporulation culture medium. SDS-PAGE, western blotting and the growth curve indicated that the best sporulation time of recombinant WB600 was 24-30 h at 37 °C with continuous shaking (250 rpm). Grass carp were fed with three levels of B.s-CotC-CP (1 × 10 6 , 1 × 10 7 , and 1 × 10 8  CFU g -1 ) incorporated in the basal pellets diet. The commercial pellets or supplemented with spores just expressing CotC (1 × 10 7  CFU g -1 ) were served as control diet. Our results showed that grass carp orally immunized with the feed-based B.s-CotC-CP developed a strong specific immune response with significantly (P sinensis in fish body. Therefore, this study demonstrated that the feed-based recombinant spores could trigger high levels of mucosal and humoral immunity, and would be a promising candidate vaccine against C. sinensis metacercariae formation in freshwater fish. Copyright © 2017 Elsevier Ltd. All rights reserved.

Multidrug resistance-associated protein 4 is a bile transporter of Clonorchis sinensis simulated by in silico docking

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Fuhong Dai

2017-11-01

Full Text Available Abstract Background Multidrug resistance-associated protein 4 (MRP4 is a member of the C subfamily of the ABC family of ATP-binding cassette (ABC transporters. MRP4 regulates ATP-dependent efflux of various organic anionic substrates and bile acids out of cells. Since Clonorchis sinensis lives in host’s bile duct, accumulation of bile juice can be toxic to the worm’s tissues and cells. Therefore, C. sinensis needs bile transporters to reduce accumulation of bile acids within its body. Results We cloned MRP4 (CsMRP4 from C. sinensis and obtained a cDNA encoding an open reading frame of 1469 amino acids. Phylogenetic analysis revealed that CsMRP4 belonged to the MRP/SUR/CFTR subfamily. A tertiary structure of CsMRP4 was generated by homology modeling based on multiple structures of MRP1 and P-glycoprotein. CsMRP4 had two membrane-spanning domains (MSD1 & 2 and two nucleotide-binding domains (NBD1 & 2 as common structural folds. Docking simulation with nine bile acids showed that CsMRP4 transports bile acids through the inner cavity. Moreover, it was found that CsMRP4 mRNA was more abundant in the metacercariae than in the adults. Mouse immune serum, generated against the CsMRP4-NBD1 (24.9 kDa fragment, localized CsMRP4 mainly in mesenchymal tissues and oral and ventral suckers of the metacercariae and the adults. Conclusions Our findings shed new light on MRPs and their homologs and provide a platform for further structural and functional investigations on the bile transporters and parasites’ survival.

Risks for fishborne zoonotic trematodes in tilapia production systems in Guangdong province, China.

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Li, Kang; Clausen, Jesper Hedegaard; Murrell, K Darwin; Liu, Liping; Dalsgaard, Anders

2013-11-15

Guangdong province is the most important region for tilapia culture in China. However, it is also an endemic region for fishborne zoonotic trematodes (FZT), which pose a risk to human food safety and health. A study was designed to assess the status of trematode parasite infections in tilapia aquaculture systems as an indicator of potential risks from FZT associated with consumption of tilapia. Tilapia from nursery and grow-out ponds were sampled from monoculture, polyculture and integrated aquaculture systems. The results from 388 tilapia examined revealed a very low prevalence (1.5%) of trematode infections (Heterophyidae and Echinostomatidae). Integrated systems using animal manure and latrine wastes as fertilizer did not show a higher prevalence of FZT. Because it was not clear whether the low risk of infection was attributable to existing effective pond management practices or a low risk of spillover of FZT from area sylvatic reservoir hosts, a survey of local wild-caught fish was conducted. Five species of FZT were discovered from a total of 271 wild-caught fish and a mean infection density of 4.0 metacercariae/100g; FZT discovered included intestinal flukes (Haplorchis spp., Procerovum varium, and Metagonimus spp.) and metacercariae tentatively identified as Clonorchis sinenesis. The common occurrence of FZT in wild-caught fish suggests that the presence of FZT in local wild animal reservoirs is substantial, and that although the current aquaculture management systems for tilapia are generally effective in preventing transmission of these parasites into tilapia production systems, the improvement of pond management practices and biosecurity must be maintained at a high level. Copyright © 2013 Elsevier B.V. All rights reserved.

Estudio del foco en un caso de fasciolosis humana en Neuquén A focus study from a case of human fascioliasis in Neuquén

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Diana Rubel

2005-06-01

Full Text Available El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222. Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2% estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10 para caprinos, 82% (9/11 para ovinos y 86% (6/7 para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5 y ovejas (4, 18, 13 infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7 (pAn epidemiological focal study was performed in Loncopué, Neuquén, Argentina, in November 2002 to detect the origin of the infection in a human case of fascioliasis confirmed by an indirect-ELISA test, six months before the study. Thirty five individual fecal samples were taken from domestic livestock, and watercress plants and snails were collected from the irrigation ditches connected to a main canal in the surroundings of the patient’s house. A new blood sample was taken from the already recovered patient. The patient was still seropositive to Fasciola hepatica antigens. No metacercariae were found in the 222 watercress leaves checked. All the snails collected (n=130 were

Monoclonal antibody against recombinant Fasciola gigantica cathepsin L1H could detect juvenile and adult cathepsin Ls of Fasciola gigantica.

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Wongwairot, Sirima; Kueakhai, Pornanan; Changklungmoa, Narin; Jaikua, Wipaphorn; Sansri, Veerawat; Meemon, Krai; Songkoomkrong, Sineenart; Riengrojpitak, Suda; Sobhon, Prasert

2015-01-01

Cathepsin Ls (CatLs), the major cysteine protease secreted by Fasciola spp., are important for parasite digestion and tissue invasion. Fasciola gigantica cathepsin L1H (FgCatL1H) is the isotype expressed in the early stages for migration and invasion. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1H (rFgCatL1H) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with recombinant proFgCatL1H (rproFgCatL1H). This MoAb is an immunoglobulin (Ig)G1 with κ light chain isotype. The MoAb reacted specifically with rproFgCatL1H, the native FgCatL1H at a molecular weight (MW) 38 to 48 kDa in the extract of whole body (WB) of metacercariae and newly excysted juvenile (NEJ) and cross-reacted with rFgCatL1 and native FgCatLs at MW 25 to 28 kDa in WB of 2- and 4-week-old juveniles, adult, and adult excretory-secretory (ES) fractions by immunoblotting and indirect ELISA. It did not cross-react with antigens in WB fractions from other parasites, including Gigantocotyle explanatum, Paramphistomum cervi, Gastrothylax crumenifer, Eurytrema pancreaticum, Setaria labiato-papillosa, and Fischoederius cobboldi. By immunolocalization, MoAb against rFgCatL1H reacted with the native protein in the gut of metacercariae and NEJ and also cross-reacted with CatL1 in 2- and 4-week-old juveniles and adult F. gigantica. Therefore, FgCatL1H and its MoAb may be used for immunodiagnosis of both early and late fasciolosis in ruminants and humans.

Epidemiology of fishborne trematodiasis in Kazakhstan.

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Sultanov, A; Abdybekova, A; Abdibaeva, A; Shapiyeva, Z; Yeshmuratov, T; Torgerson, P R

2014-10-01

Fish borne trematodes are an important cause of morbidity in Kazakhstan. The number of human cases of opisthorchiidosis (infection with parasites of the family opisthorchiidae) reached a peak of 2521 recorded cases (17 cases per 100,000) in 2002 with a gradual decline to 1225 cases (7.4 cases per 100,000) in 2011. Most human cases are found in the north and north east part of Kazakhstan in areas drained by the Irtysh River and it tributaries. A further focus is found in the north west in the Ural river basin in the European part of Kazakhstan. The most common occupations of patients with opisthorchiidosis included the manual laborers, those employed in the home or unemployed. Necropsy investigations of village dogs in an endemic region revealed 37 of 51 (72%) village dogs infected with either Opisthorchis felineus or Methorchis bilis. Likewise an investigation of 242 cats consisting of strays, village, suburban and city cats revealed 79 (33%) animals infected with O. felineus. Higher prevalences were seen in strays and village cats compared to suburban cats. No urban cats, which lived in apartments, were found to be infected. Other important zoonoses included Echinococcus granulosus, detected in 2 of the 51 necropsied village dogs and E. multilocularis was found in 2 out of 124 necropsied stray cats. Investigations of locally caught fish revealed 10 of 107 (9%) roach (Rutilus rutilus), 49 of 68(72%) ide (Leuciscus idus) and 2 of 79 (2.5%) bream (Abramis brama) infested with trematode metacercariae. No metacercariae were found in 609 crucian carp (Carassius carassius), 35 tench (Tinca tinca), 79 carp (Cyprinus carpio), 46 perch (Perca fluviatilis) or 20 zander (Sander lucioperca). Copyright © 2014 Elsevier B.V. All rights reserved.

A Bayesian analysis of the parasitic ecology in Jenynsia multidentata(Pisces: Anablepidae

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Martin M. Montes

2017-09-01

Full Text Available ABSTRACT Jenynsia multidentata Jenyns, 1842 (one-sided livebearers are euryhaline viviparous fish of small size, used in the laboratory experiment, important as resource for biological control of mosquito’s larva and a key species to recover eutrophic lakes. Works have been published dealing with parasite biodiversity of this host, but little has been studied about the parasite community ecology. From early 2009 to ends of 2010 specimens of J. multidentata were collected from two places, the Salado Relief Channel (S.R.C. on Samborombón Bay and the Sauce Chico River near to the city of Bahia Blanca (B.B.. All fish were sexed, measured and grouped into sizes/age classes. The fishes from both sites harbored 16 parasitic species: nine digenean, one monogenean, one metacestode, one acanthocephalan, two nematode and two copepods. Lecithaster confusus Odhner, 1905, the metacercariae Hemiuridae gen. sp. indet., metacercariae Thylodelphys sp. (inside the eye, Glossocercus sp. nematode L4 (intestine and Ergasilus sieboldii Nordmann, 1832 are new records for the host. The high number of larval stages made of this fish a link between micro and macroecosystems. The size 2 had the higher biodiversity in both sites, which could be the most suitable age to store the maximal number of parasite of the environment and have a more equitability in their distribution on the host. In B.B. some parasites had higher prevalence and mean abun dance due to the small size of the waterbody compared with the S.R.C. Despite that, in S.R.C. exist a higher specific richness and biodiversity due the daily flow of saline and freshwater and proximity of the sample site to the mouth of the channel in the bay. This is the first approximation to an analysis of the parasitic ecology on this host.

Molecular characterization of severin from Clonorchis sinensis excretory/secretory products and its potential anti-apoptotic role in hepatocarcinoma PLC cells.

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Xueqing Chen

Full Text Available BACKGROUND: Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis, is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC. It has been well known that the excretory/secretory products of C. sinensis (CsESPs play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin, which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. METHODOLOGY/PRINCIPAL FINDINGS: There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. CONCLUSIONS/SIGNIFICANCE: Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the

Sequence analysis and characterization of pyruvate kinase from Clonorchis sinensis, a 53.1-kDa homopentamer, implicated immune protective efficacy against clonorchiasis

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Tingjin Chen

2017-11-01

Full Text Available Abstract Background Clonorchis sinensis, the causative agent of clonorchiasis, is classified as one of the most neglected tropical diseases and affects more than 15 million people globally. This hepatobiliary disease is highly associated with cholangiocarcinoma. As key molecules in the infectivity and subsistence of trematodes, glycolytic enzymes have been targets for drug and vaccine development. Clonorchis sinensis pyruvate kinase (CsPK, a crucial glycolytic enzyme, was characterized in this research. Results Differences were observed in the sequences and spatial structures of CsPK and PKs from humans, rats, mice and rabbits. CsPK possessed a characteristic active site signature (IKLIAKIENHEGV and some unique sites but lacked the N-terminal domain. The predicted subunit molecular mass (Mr of CsPK was 53.1 kDa. Recombinant CsPK (rCsPK was a homopentamer with a Mr. of approximately 290 kDa by both native PAGE and gel filtration chromatography. Significant differences in the protein and mRNA levels of CsPK were observed among four life stages of C. sinensis (egg, adult worm, excysted metacercaria and metacercaria, suggesting that these developmental stages may be associated with diverse energy demands. CsPK was widely distributed in adult worms. Moreover, an intense Th1-biased immune response was persistently elicited in rats immunized with rCsPK. Also, rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. Conclusions The sequences and spatial structures, molecular mass, and expression profile of CsPK have been characterized. rCsPK was indicated to be a homopentamer. Rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. CsPK is worthy of further study as a promising target for drug and vaccine development.

Molecular Characterization of Severin from Clonorchis sinensis Excretory/Secretory Products and Its Potential Anti-apoptotic Role in Hepatocarcinoma PLC Cells

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He, Lei; Wang, Xiaoyun; Liang, Pei; Chen, Wenjun; Bian, Meng; Ren, Mengyu; Lin, Jinsi; Liang, Chi; Xu, Jin; Wu, Zhongdao; Li, Xuerong; Huang, Yan; Yu, Xinbing

2013-01-01

Background Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis), is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC). It has been well known that the excretory/secretory products of C. sinensis (CsESPs) play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin), which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. Methodology/Principal Findings There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin) and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. Conclusions/Significance Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the process of HCC

Developmental Transcriptomic Features of the Carcinogenic Liver Fluke, Clonorchis sinensis

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Cho, Pyo Yun; Kim, Tae Im; Cho, Shin-Hyeong; Choi, Sang-Haeng; Park, Hong-Seog; Kim, Tong-Soo; Hong, Sung-Jong

2011-01-01

Clonorchis sinensis is the causative agent of the life-threatening disease endemic to China, Korea, and Vietnam. It is estimated that about 15 million people are infected with this fluke. C. sinensis provokes inflammation, epithelial hyperplasia, and periductal fibrosis in bile ducts, and may cause cholangiocarcinoma in chronically infected individuals. Accumulation of a large amount of biological information about the adult stage of this liver fluke in recent years has advanced our understanding of the pathological interplay between this parasite and its hosts. However, no developmental gene expression profiles of C. sinensis have been published. In this study, we generated gene expression profiles of three developmental stages of C. sinensis by analyzing expressed sequence tags (ESTs). Complementary DNA libraries were constructed from the adult, metacercaria, and egg developmental stages of C. sinensis. A total of 52,745 ESTs were generated and assembled into 12,830 C. sinensis assembled EST sequences, and then these assemblies were further categorized into groups according to biological functions and developmental stages. Most of the genes that were differentially expressed in the different stages were consistent with the biological and physical features of the particular developmental stage; high energy metabolism, motility and reproduction genes were differentially expressed in adults, minimal metabolism and final host adaptation genes were differentially expressed in metacercariae, and embryonic genes were differentially expressed in eggs. The higher expression of glucose transporters, proteases, and antioxidant enzymes in the adults accounts for active uptake of nutrients and defense against host immune attacks. The types of ion channels present in C. sinensis are consistent with its parasitic nature and phylogenetic placement in the tree of life. We anticipate that the transcriptomic information on essential regulators of development, bile chemotaxis, and

Sequence analysis and characterization of pyruvate kinase from Clonorchis sinensis, a 53.1-kDa homopentamer, implicated immune protective efficacy against clonorchiasis.

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Chen, Tingjin; Jiang, Hongye; Sun, Hengchang; Xie, Zhizhi; Ren, Pengli; Zhao, Lu; Dong, Huimin; Shi, Mengchen; Lv, Zhiyue; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing; Huang, Yan; Xu, Jin

2017-11-09

Clonorchis sinensis, the causative agent of clonorchiasis, is classified as one of the most neglected tropical diseases and affects more than 15 million people globally. This hepatobiliary disease is highly associated with cholangiocarcinoma. As key molecules in the infectivity and subsistence of trematodes, glycolytic enzymes have been targets for drug and vaccine development. Clonorchis sinensis pyruvate kinase (CsPK), a crucial glycolytic enzyme, was characterized in this research. Differences were observed in the sequences and spatial structures of CsPK and PKs from humans, rats, mice and rabbits. CsPK possessed a characteristic active site signature (IKLIAKIENHEGV) and some unique sites but lacked the N-terminal domain. The predicted subunit molecular mass (Mr) of CsPK was 53.1 kDa. Recombinant CsPK (rCsPK) was a homopentamer with a Mr. of approximately 290 kDa by both native PAGE and gel filtration chromatography. Significant differences in the protein and mRNA levels of CsPK were observed among four life stages of C. sinensis (egg, adult worm, excysted metacercaria and metacercaria), suggesting that these developmental stages may be associated with diverse energy demands. CsPK was widely distributed in adult worms. Moreover, an intense Th1-biased immune response was persistently elicited in rats immunized with rCsPK. Also, rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. The sequences and spatial structures, molecular mass, and expression profile of CsPK have been characterized. rCsPK was indicated to be a homopentamer. Rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. CsPK is worthy of further study as a promising target for drug and vaccine development.

Identification, sequence analysis, and characterization of serine/threonine protein kinase 17A from Clonorchis sinensis.

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Huang, Lisi; Lv, Xiaoli; Huang, Yan; Hu, Yue; Yan, Haiyan; Zheng, Minghui; Zeng, Hua; Li, Xuerong; Liang, Chi; Wu, Zhongdao; Yu, Xinbing

2014-05-01

This is the first report of a novel protein from Clonorchis sinensis (C. sinensis), serine/threonine protein kinase 17A (CsSTK17A), which belongs to a member of the death-associated protein kinase (DAPK) family known to regulate diverse biological processes. The full-length sequence encoding CsSTK17A was isolated from C. sinensis adult cDNA plasmid library. Two transcribed isoforms of the gene were identified from the genome of C. sinensis. CsSTK17A contains a kinase domain at the N-terminus that shares a degree of conservation with the DAPK families. Besides, the catalytic domain contains 11 subdomains conserved among STKs and shares the highest identity with STK from Schistosoma mansoni (55.9%). Three-dimensional structure of CsSTK17A displays the canonical STK fold, including the helix C, P-loop, and the activation loop. We obtained recombinant CsSTK17A (rCsSTK17A) and anti-rCsSTK17A IgG. The rCsSTK17A could be probed by anti-rCsSTK17A rat serum, C. sinensis-infected rat serum and the sera from rats immunized with C. sinensis excretory-secretory products, indicating that it is a circulating antigen possessing a strong immunocompetence. Moreover, quantitative RT-PCR and western blotting analyses revealed that CsSTK17A exhibited the highest mRNA and protein expression level in eggs, followed by metacercariae and adult worms. Intriguingly, in the immunolocalization assay, CsSTK17A was intensively localized to the operculum region of eggs in uterus, as well as the vitelline gland of both adult worm and metacercaria, implying that the protein was associated with the reproduction and development of C. sinensis. Overall, these fundamental studies might contribute to further researches on signaling systems of the parasite.

Developmental transcriptomic features of the carcinogenic liver fluke, Clonorchis sinensis.

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Won Gi Yoo

2011-06-01

Full Text Available Clonorchis sinensis is the causative agent of the life-threatening disease endemic to China, Korea, and Vietnam. It is estimated that about 15 million people are infected with this fluke. C. sinensis provokes inflammation, epithelial hyperplasia, and periductal fibrosis in bile ducts, and may cause cholangiocarcinoma in chronically infected individuals. Accumulation of a large amount of biological information about the adult stage of this liver fluke in recent years has advanced our understanding of the pathological interplay between this parasite and its hosts. However, no developmental gene expression profiles of C. sinensis have been published. In this study, we generated gene expression profiles of three developmental stages of C. sinensis by analyzing expressed sequence tags (ESTs. Complementary DNA libraries were constructed from the adult, metacercaria, and egg developmental stages of C. sinensis. A total of 52,745 ESTs were generated and assembled into 12,830 C. sinensis assembled EST sequences, and then these assemblies were further categorized into groups according to biological functions and developmental stages. Most of the genes that were differentially expressed in the different stages were consistent with the biological and physical features of the particular developmental stage; high energy metabolism, motility and reproduction genes were differentially expressed in adults, minimal metabolism and final host adaptation genes were differentially expressed in metacercariae, and embryonic genes were differentially expressed in eggs. The higher expression of glucose transporters, proteases, and antioxidant enzymes in the adults accounts for active uptake of nutrients and defense against host immune attacks. The types of ion channels present in C. sinensis are consistent with its parasitic nature and phylogenetic placement in the tree of life. We anticipate that the transcriptomic information on essential regulators of development

HISTOPATOLOGIA DO PARASITISMO POR METACERCARIAS DE Clinostomum sp EM TAMOATA, Hoplosternum littorale (HANCOCK, 1828)

OpenAIRE

Carmona de Sao Clemente, Sergio; Matos, Edilson; Tortelly, Rogerio; de Lima, Francisco Carlos

1998-01-01

Foi realizado um estudo enfocando o aspecto histopatológico provocado por metacercárias do género Clinostomum em 26 espécimes de Hoplosternum littorale coletados no Rio Guamá, Cidade de Belém, Pará 85% dos peixes apresentaram o parasitismo. No exame microscópico, ao redor de formaçães císticas contendo o parasito, observou-se fibras musculares em degeneração ao lado de moderada reação inflamatória com infiltração de células mononucleares. O quadro apresentado é sugestivo de miosite parasitári...

Relative effects of temperature, light, and humidity on clinging behavior of metacercariae-infected ants

DEFF Research Database (Denmark)

Botnevik, C.F.; Malagocka, Joanna; Jensen, Annette Bruun

2016-01-01

The lancet fluke, Dicrocoelium dendriticum, is perhaps the best-known example of parasite manipulation of host behavior, which is manifested by a radically changed behavior that leaves infected ants attached to vegetation at times when transmission to an herbivore host is optimal. Despite...... the publicity surrounding this parasite, curiously little is known about factors inducing and maintaining behavioral changes in its ant intermediate host. This study examined the importance of 3 environmental factors on the clinging behavior of red wood ants, Formica polyctena , infected with D. dendriticum...... . This behavior, hypothesized to involve cramping of the mandibular muscles in a state of tetany, was observed in naturally infected F. polyctena under controlled temperature, light, and humidity conditions. We found that low temperature significantly stimulated and maintained tetany in infected ants while light...

[Experimental and epidemiological studies on the life cycle of Echinostoma hortense Asada, 1926 (Trematoda: Echinostomatidae)

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Ahn, Yung Kyum; Ryang, Yong Suk

1986-12-01

Recently there have been some reports on human infections of Echinostoma hortense in Korea. It was found that a few species of freshwater fishes were playing the role of the second intermediate host of E. hortense. However, molluscan intermediate host has not been identified yet in Korea. The present study aimed to establish the life cycle of E. hortense in laboratory. Experimental studies such as egg production from the rat, development of the eggs in vitro, exposure of miracidia to freshwater snails, shedding pattern of cercariae from infected snails, morphology of cercariae, cercarial infection to the second intermediate host and infection of metacercariae to the difinitive hosts were done. In addition, epidemiological surveys on the infection status in inhabitants and house rats, and on the natural infection of larval echinostomes in the snails and fishes were carried out along the South Hangang-river. The results obtained were as follows: The eggs deposited from adults in physiological saline were cultivated at room temperature (20-24C). The miracidia were firstly observed on 8 days after cultivation, and 85.5 per cent of the eggs contained the mature miracidia on 11 days after cultivation. More than 90 per cent formed the miracidia when cultivated at temperature 22-27C. Hatching of the miracidia began on 12 days after cultivation and continued for a week. The size of the miracidia was 103.0 x 51.4 micrometer in average. The motility of miracidia were active up to 8 hours after shedding, but they were all dead within 10 hours after shedding. A freshwater snail, Radix auricularia coreana was cultivated in aquaria. A hatched F1 snails from the egg masses were exposed to 20 miracidia respectively. Escape of cercariae started on 15 days after infection. Radix auricularia coreana was experimentally identified as the first intermediate host of E. hortense in Korea. Cercarial shedding started on 15-20 days after infection by snail, continued for about 10 days (8

European distribution for metacercariae of the North American digenean Posthodiplostomum cf. minimum centrarchi (Strigeiformes: Diplostomidae)

Czech Academy of Sciences Publication Activity Database

Kvach, Yuriy; Jurajda, Pavel; Bryjová, Anna; Trichkova, T.; Ribeiro, F.; Přikrylová, I.; Ondračková, Markéta

2017-01-01

Roč. 66, č. 5 (2017), s. 635-642 ISSN 1383-5769 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:68081766 Keywords : Lepomis gibbosus * Micropterus salmoides * White grub * Physid snails * Non-indigenous species Subject RIV: EG - Zoology OBOR OECD: Parasitology Impact factor: 1.744, year: 2016

Production and characterization of a monoclonal antibody against recombinant cathepsin L1 of Fasciola gigantica.

Science.gov (United States)

Anuracpreeda, Panat; Srirakam, Thippawan; Pandonlan, Sudarat; Changklungmoa, Narin; Chotwiwatthanakun, Charoonroj; Tinikul, Yotsawan; Poljaroen, Jaruwan; Meemon, Krai; Sobhon, Prasert

2014-08-01

Monoclonal antibodies (MoAbs) against a recombinant cathepsin L1 of Fasciola gigantica (rFgCatL1) were produced in vitro by fusion of BALB/c mice spleen cells immunized with rFgCatL1 and mouse myeloma cells. Reactivity and specificity of these MoAbs were evaluated by indirect ELISA and immunoblotting techniques. Seven MoAb clones were selected from the stable hybridoma clones, namely 1E10, 1F5, 3D11, 4B10, 4D3, 4E3 and 5E7. Clones 1E10, 1F5 and 3D11 were IgM, whereas clones 4B10, 4D3, 4E3 and 5E7 were IgG1. All MoAbs had kappa light chain isotypes. All MoAbs reacted with rCatL1 at molecular weight (MW) 30kDa and with the native CatL1 at MW 27kDa in whole body (WB) extracts of metacercariae (Met), newly excysted juveniles (NEJ), 1, 3, 5-week-old juveniles (Ju), adult WB and adult excretory-secretory (ES) fractions, but not with adult tegumental antigens (TA). All of these MoAbs showed no cross-reactions with antigens of other parasites commonly found in ruminants and human, including Paramphistomum cervi, Eurytrema pancreaticum, Gigantocotyle explanatum, Schistosoma spindale, Schistosoma mansoni, Moniezia benedeni, Avitellina centripunctata, Trichuris sp., Haemonchus placei and Setaria labiato-papillosa. Localization of CatL1 in each developmental stages of F. gigantica by immunoperoxidase technique, using these MoAbs as probes, indicated that CatL1 was present at high concentration in the caecal epithelium and caecal lumen of metacercariae, NEJ, 1, 3, 5-week-old juveniles and adult fluke. This finding indicated that CatL1 is a copiously expressed parasite protein that is released into the ES, thus CatL1 and its MoAb could be a good candidate for immunodiagnosis of fasciolosis in ruminant and human. Copyright © 2014 Elsevier B.V. All rights reserved.

Gene Expression Profile in the Liver of BALB/c Mice Infected with Fasciola hepatica.

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Jose Rojas-Caraballo

Full Text Available Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs. It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with F. hepatica metacercariae using a microarray-based methodology.A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0, seven days post-infection (t7 and twenty-one days post-infection (t21.We found that F. hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death.The present study provides us insights at the molecular level about the underlying mechanisms used by F. hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma

Gene Expression Profile in the Liver of BALB/c Mice Infected with Fasciola hepatica.

Science.gov (United States)

Rojas-Caraballo, Jose; López-Abán, Julio; Fernández-Soto, Pedro; Vicente, Belén; Collía, Francisco; Muro, Antonio

2015-01-01

Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs). It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with F. hepatica metacercariae using a microarray-based methodology. A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain) weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0), seven days post-infection (t7) and twenty-one days post-infection (t21). We found that F. hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death. The present study provides us insights at the molecular level about the underlying mechanisms used by F. hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma. However

Sequence analysis and molecular characterization of Clonorchis sinensis hexokinase, an unusual trimeric 50-kDa glucose-6-phosphate-sensitive allosteric enzyme.

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Tingjin Chen

Full Text Available Clonorchiasis, which is induced by the infection of Clonorchis sinensis (C. sinensis, is highly associated with cholangiocarcinoma. Because the available examination, treatment and interrupting transmission provide limited opportunities to prevent infection, it is urgent to develop integrated strategies to prevent and control clonorchiasis. Glycolytic enzymes are crucial molecules for trematode survival and have been targeted for drug development. Hexokinase of C. sinensis (CsHK, the first key regulatory enzyme of the glycolytic pathway, was characterized in this study. The calculated molecular mass (Mr of CsHK was 50.0 kDa. The obtained recombinant CsHK (rCsHK was a homotrimer with an Mr of approximately 164 kDa, as determined using native PAGE and gel filtration. The highest activity was obtained with 50 mM glycine-NaOH at pH 10 and 100 mM Tris-HCl at pH 8.5 and 10. The kinetics of rCsHK has a moderate thermal stability. Compared to that of the corresponding negative control, the enzymatic activity was significantly inhibited by praziquantel (PZQ and anti-rCsHK serum. rCsHK was homotropically and allosterically activated by its substrates, including glucose, mannose, fructose, and ATP. ADP exhibited mixed allosteric effect on rCsHK with respect to ATP, while inorganic pyrophosphate (PPi displayed net allosteric activation with various allosteric systems. Fructose behaved as a dose-dependent V activator with the substrate glucose. Glucose-6-phosphate (G6P displayed net allosteric inhibition on rCsHK with respect to ATP or glucose with various allosteric systems in a dose-independent manner. There were differences in both mRNA and protein levels of CsHK among the life stages of adult worm, metacercaria, excysted metacercaria and egg of C. sinensis, suggesting different energy requirements during different development stages. Our study furthers the understanding of the biological functions of CsHK and supports the need to screen for small

Sequence Analysis and Molecular Characterization of Clonorchis sinensis Hexokinase, an Unusual Trimeric 50-kDa Glucose-6-Phosphate-Sensitive Allosteric Enzyme

Science.gov (United States)

Chen, Tingjin; Ning, Dan; Sun, Hengchang; Li, Ran; Shang, Mei; Li, Xuerong; Wang, Xiaoyun; Chen, Wenjun; Liang, Chi; Li, Wenfang; Mao, Qiang; Li, Ye; Deng, Chuanhuan; Wang, Lexun; Wu, Zhongdao; Huang, Yan; Xu, Jin; Yu, Xinbing

2014-01-01

Clonorchiasis, which is induced by the infection of Clonorchis sinensis (C. sinensis), is highly associated with cholangiocarcinoma. Because the available examination, treatment and interrupting transmission provide limited opportunities to prevent infection, it is urgent to develop integrated strategies to prevent and control clonorchiasis. Glycolytic enzymes are crucial molecules for trematode survival and have been targeted for drug development. Hexokinase of C. sinensis (CsHK), the first key regulatory enzyme of the glycolytic pathway, was characterized in this study. The calculated molecular mass (Mr) of CsHK was 50.0 kDa. The obtained recombinant CsHK (rCsHK) was a homotrimer with an Mr of approximately 164 kDa, as determined using native PAGE and gel filtration. The highest activity was obtained with 50 mM glycine-NaOH at pH 10 and 100 mM Tris-HCl at pH 8.5 and 10. The kinetics of rCsHK has a moderate thermal stability. Compared to that of the corresponding negative control, the enzymatic activity was significantly inhibited by praziquantel (PZQ) and anti-rCsHK serum. rCsHK was homotropically and allosterically activated by its substrates, including glucose, mannose, fructose, and ATP. ADP exhibited mixed allosteric effect on rCsHK with respect to ATP, while inorganic pyrophosphate (PPi) displayed net allosteric activation with various allosteric systems. Fructose behaved as a dose-dependent V activator with the substrate glucose. Glucose-6-phosphate (G6P) displayed net allosteric inhibition on rCsHK with respect to ATP or glucose with various allosteric systems in a dose-independent manner. There were differences in both mRNA and protein levels of CsHK among the life stages of adult worm, metacercaria, excysted metacercaria and egg of C. sinensis, suggesting different energy requirements during different development stages. Our study furthers the understanding of the biological functions of CsHK and supports the need to screen for small molecule inhibitors

Comments on human eurytremiasis in Brazil.

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Pinto, Hudson Alves; de Melo, Alan Lane

2016-05-20

Eurytremiasis is an important parasitic disease of cattle that was recently suggested to be a neglected and emerging human disease in Brazil. Based on a misinterpretation of the life cycle of the parasite, it was suggested that a great number of people could be infected with this fluke in the country. In the present letter, aspects of the life cycle of Eurytrema spp. are revisited and clarified. The mechanism of transmission previously reported for the few accidental human cases involved the ingestion of raw or undercooked insects (grasshoppers and crickets) harboring the infective metacercariae. In reality, the zoonotic potential of Eurytrema species is extremely low, and human eurytremiasis is not, and probably never will be, a zoonotic disease in countries where entomophagy is not a common food habit.

Parasites of juvenile golden grey mullet Liza aurata Risso, 1810 in Sarıkum Lagoon Lake at Sinop, Turkey.

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Öztürk, Türkay

2013-12-01

Juvenile golden grey mullet, Liza aurata were collected from Sarıkum Lagoon Lake which connected to the Black Sea at Sinop, Turkey and examined for parasitic fauna. A total of 219 fish were investigated throughout a 1-year period. Parasite species recovered were Trichodina lepsii, T. puytoraci, Gyrodactylus sp., Ligophorus cephali, Ligophorus mediterraneus, Solostamenides mugilis, Ascocotyle sp. (metacercaria) and Ergasilus lizae. Overall infection prevalence (%) and mean intensity values were 95.9% and 412.65 ± 85.31 parasites per infected fish, respectively. Infection prevalence and mean intensity values for each parasite species in relation to season and fish size were also determined and discussed. While Ligophorus cephali and L. mediterraneus are new records for Turkish parasite fauna, the juvenile Liza aurata is a new host record for Ligophorus cephali and L. mediterraneus.

Experimental infection with the small intestinal trematode, Haplorchis pumilio, in young dogs

DEFF Research Database (Denmark)

Nissen, Sofie; Nguyen, Lan Anh Thi; Dalsgaard, Anders

2013-01-01

included as uninfected controls. Faecal examination for eggs was performed twice weekly using a sieving and sedimentation technique. Body temperature and weight of the dogs were measured as was total white blood cells, blood eosinophils and packed cell volume. Subsets of dogs were examined post......Fishborne zoonotic trematodes (FZT) are highly prevalent in Southeast Asia. Recent studies on the role of domestic animals in the transmission of FZT in Northern Vietnam found that dogs, mainly infected with Haplorchis pumilio, contributed widely to the transmission of FZT. On this background, we...... conducted an experimental infection with H. pumilio to elucidate population dynamics and host reactions in dogs. Eight household-reared dogs (3-6 months old), were each orally infected with 500 H. pumilio metacercariae obtained by artificial digestion of naturally infected fish. Another eight dogs were...

Time-course investigation of the gene expression profile during Fasciola hepatica infection: A microarray-based study

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Jose Rojas-Caraballo

2015-12-01

Full Text Available Fasciolosis is listed as one of the most important neglected tropical diseases according with the World Health Organization and is also considered as a reemerging disease in the human beings. Despite there are several studies describing the immune response induced by Fasciola hepatica in the mammalian host, investigations aimed at identifying the expression profile of genes involved in inducing hepatic injury are currently scarce. Data presented here belong to a time-course investigation of the gene expression profile in the liver of BALB/c mice infected with F. hepatica metacercariae at 7 and 21 days after experimental infection. The data published here have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE69588, previously published by Rojas-Caraballo et al. (2015 in PLoS One [1].

Helminths Parasite Larvae collected from Arabian Gulf Fish. 4. Description of four Larvae including two Metacercarae, one Didymozoid and one Acanthocephalan from Emirati Coasts

International Nuclear Information System (INIS)

Kardousha, M.M.

2005-01-01

Four helminth larvae from different fish hosts caught from Emirate coasts are described. Two are metacercariae related to genus Southampton's, type (I) and (II). Type (I) was found encysted in body cavities of the Indian halibut Psettodes erumei and the areolate grouper Epinephelus areolatusu and type (II) in the golden stripped goatfish Mulloides flavolineatus. The third larva is related to Acanthocephala and identified as Serrasentis sagittifer. It was collected from the body cavity of the spotted lizard fish Saurida undosquamius. The fourth larva belongs to the didymozoid trematodes, and was found infecting the kidneys of different hosts, such as Saurida undosquamis, the Jack pomfret Parastromateus niger and mackerel tuna Euthynnus affins. The larva related to Stephanostomum type (I) and type (II) and the dydimozoid type are described for the first time in the Arabian Gulf. (author)

Trematodiasis en algunos peces del lago de Cuitzeo, Michoacán, México

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Ma. del Carmen Guzmán-Cornejo

1999-09-01

Full Text Available In September, 1995 and January, 1996, six species of fishes from Lake Cuitzeo, Michoacan, Mexico, were collected (Alloophorus robustus n=30, Goodea atripinnis n=30, Xenotoca variata n=41, Chirostoma jordani n=30, Carassius auratus n=30 and Oreochromis aureus n=30, to establish the infection levels reached by Posthodiplostomum minimum, Clinostomum complanatum and Diplostomum (Tylodelphys sp., in their hosts. Infection caused by P. minimum metacercariae was the most important helminthiasis considering the number of hosts they parasite (five of six, and the high levels of abundance of the infection; we suggest that fish preferences and the generalist character of the trematode are the causes that determine this infection. Finally, the role of goodeids and atherinids in transmission of infections in the lake is confirmed; the former being the main group mantaining such infections.

Hepatobiliary Fascioliasis: Clinical and Radiological Features

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K Aghazade

2007-08-01

Full Text Available Fascioliasis is a worldwide but unevenly distributed zoonosis caused by the trematode Fasciola hepatica that infects do­mesti­cated herbivores. Fasciolosis also occurs accidentally in humans by ingestion of metacercaria-laden freshwater or water plants. Human infections are common in developing countries and are not rare in Europe. The clinical course has been con­ventionally described in two phases: an acute phase of hepatic parenchymal invasion of an immature worm larva (parenchy­mal phase and a stationary phase after residence in the bile duct and production of eggs (ductal phase. We report a 34 years old woman from Ilam, western Iran with hepatic disorder, RUQ pain, and jaundice. The diagnosis was made by sonography, CT scan and serologic studies. Serologic exam (ELISA was positive & CT findings were compatible with fascioliasis.

Parasites of the mink frog (rana septentrionalis) from Minnesota, U.S.A.

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Schotthoefer, Anna M.; Bolek, M.G.; Cole, Rebecca A.; Beasley, Val R.

2009-01-01

Twenty-two mink frogs, Rana septentrionalis, collected from two locations in Minnesota, United States, were examined for helminth and protozoan blood parasites in July 1999. A total of 16 parasite taxa were recovered including 5 larval digenean trematodes, 7 adult digenean trematodes, 3 nematodes, and I Trypanosorna species. Infracommunities were dominated by the digeneans in terms of richness and abundance. In particular, echinostomatid metacercariae in the kidneys of frogs were the most common parasites found, infecting 100% of the frogs and consisting of about 90% of all helminth individuals recovered. Gorgodera amplicava, Gorgoderina multilohata, Haernaroloechus pan'iplexus, Haernatoloechus breviplexus, Cosnwcercoides dukae, and Oswaldocruzia pipiens represent new host records. The survey presented here represents the second known helminth survey of mink frogs conducted in North America. A summary of metazoan parasites reported from mink frogs is included.

Seasonal dynamics of Posthodiplostomum cuticola (Digenea, Diplostomatidae) metacercariae and parasite-enhanced growth of juvenile host fish

Czech Academy of Sciences Publication Activity Database

Ondračková, Markéta; Reichard, Martin; Jurajda, Pavel; Gelnar, M.

2004-01-01

Roč. 93, č. 2 (2004), s. 131-136 ISSN 0932-0113 R&D Projects: GA ČR GA524/02/0924 Institutional research plan: CEZ:AV0Z6093917 Keywords : black-spot disease * juvenile fish Subject RIV: EG - Zoology Impact factor: 1.060, year: 2004

Epidemiological studies of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe

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D.M. Pfukenyi

2006-09-01

Full Text Available During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000 in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16 264 (calves 5 418; weaners 5 461 and adults 5 385 faecal samples were collected during the entire period of the study and 2 500 (15.4 % of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001, for adult cattle than calves ( P < 0.01 and in the wet season over the dry season (P < 0.01. Faecal egg output peaked from August / September to March / April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P < 0.01. The snail population was low between December and March and started

Epidemiological studies of amphistome infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe

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D.M. Pfukenyi

2005-09-01

Full Text Available During the period between January 1999 and December 2000, the distribution and seasonal patterns of amphistome infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas, respectively. Patterns of distribution and seasonal fluctuations of intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000 in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of amphistome metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of amphistomes. A total of 16 264 (calves 5 418, weaners 5 461 and adults 5 385 faecal samples were collected during the entire period of the study and 4 790 (29.5 % of the samples were positive for amphistome eggs. For both regions the number of animals positive for amphistome eggs differed significantly between the 2 years, with the second year having a significantly higher prevalence (P < 0.01 than the first year. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001, for adult cattle than calves (P < 0.01, and in the wet over the dry season (P < 0.01. Faecal egg output peaked from October to March in both years of the study. Bulinus tropicus, Bulinus forskalii and Biomphalaria pfeifferi were recorded from the study sites. The main

Molecular cloning, characterization and functional analysis of a novel juvenile-specific cathepsin L of Fasciola gigantica.

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Sansri, Veerawat; Changklungmoa, Narin; Chaichanasak, Pannigan; Sobhon, Prasert; Meemon, Krai

2013-10-01

Cathepsin L proteases are a major class of endopeptidases expressed at a high level in Fasciola parasites. Several isoforms of cathepsin L were detected and they may perform different functions during the parasite development. In this study, a complete cDNA encoding a cathepsin L protease was cloned from a newly excysted juvenile (NEJ) cDNA library of Fasciola gigantica and named FgCatL1H. It encoded a 326 amino acid preproenzyme which shared 62.8-83.1% and 39.5-42.9% identity to Fasciola spp. and mammalian cathepsins L, respectively. All functionally important residues previously described for cathepsin L were conserved in FgCatL1H. Phylogenetic analysis demonstrated that FgCatL1H belonged to a distinct group, clade 4, with respect to adult and other juvenile Fasciola cathepsin L genes. FgCatL1H expression was detected by RT-PCR, using gene specific primers, in metacercariae and NEJ, and the expression gradually decreased in advanced developmental stages. A recombinant proFgCatL1H (rproFgCatL1H) was expressed in the yeast Pichia pastoris, affinity purified, and found to migrate in SDS-PAGE at approximately 47.6 and 38.3kDa in glycosylated and deglycosylated forms, respectively. The molecular mass of the activated mature rFgCatL1H in glycosylated form was approximately 40.7kDa. Immunoblotting and immunohistochemistry using rabbit antibodies against rproFgCatL1H showed that FgCatL1H was predominantly expressed in epithelial cells of the digestive tract of metacercariae, NEJs and juveniles of F. gigantica. FgCatL1H could cleave the synthetic fluorogenic substrate Z-Phe-Arg-MCA preferentially over Z-Gly-Pro-Arg-MCA at an optimum pH of 6.5. It also showed hydrolytic activity against native substrates, including type I collagen, laminin, and immunoglobulin G (IgG) in vitro, suggesting possible roles in host tissue migration and immune evasion. Therefore, the FgCatL1H is a possible target for vaccine and chemotherapy for controlling F. gigantica infection. Copyright �

An epidemiological study of pleuropulmonary paragonimiasis among pupils in the peri-urban zone of Kumba town, Meme Division, Cameroon

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Kefie-Arrey Charles

2003-12-01

Full Text Available Abstract Background Paragonimiasis have previously been reported in two zones of the Southwest Province of Cameroon including the Kupe mountain and Mundani foci. The aim of this study was to investigate the presence and epidemiology of paragonimiasis in the peri-urban zone of Kumba, Meme Division, located about 50 km away from the Kupe mountain focus. Methods Pupils of several government primary schools in 5 villages around Kumba underwent both parasitologic and clinical investigations in search of signs and symptoms of paragonimiasis. Mycobacterium tuberculosis was also searched for in the differential diagnosis.Freshwater crabs from neighbouring streams in the five villages were dissected in search of paragonimus metacercariae. Results Out of a total of 1482 pupils examined in all five villages, 309 individuals (147 males and 162 females were recruited for this study based on the presence of one or more signs or symptoms of paragonimiasis. Eggs of Paragonimus africanus were found in stools and/or sputum of pupils from all five villages, giving an overall paragonimus prevalence of 2.56%. There was no significant difference in the disease prevalence between the villages (X2 = 8.36, P = 0.08. The prevalence of Paragonimus africanus eggs amongst pupils with symptoms of paragonimiasis was 12.3% (38 of 309. Males were infected more than females (17.0% versus 8.0%, but the difference was not significant (X2 = 5.76, P = 0.16. All the 38 paragonimus egg positive subjects presented with cough, 23 (60.53% complained of chest pain while 16 (42.11% had haemoptysis. Stool examinations also detected some intestinal parasites including Ascaris lumbricoides (29.45%, Trichuris trichiura (6.47%, Necator americanus (2.27%, Strongyloides stercoralis (1.62%, Enterobius vermicularis (0.65%, and Entamoeba histolytica (4.53%. No case of M. tuberculosis was noted. Out of a total of 85 dissected crabs (Sudanonautes africanus, 6.02 % were infected with paragonimus

Ocorrência de infecção natural de Fasciola hepatica Linnaeus, 1758 em Lymnaea columella Say, 1817 no Vale do Paraíba, SP, Brasil Natural infection by Fasciola hepatica in Lymnaea columella in the Paraíba river valley, S. Paulo, Brazil

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Marlene Tiduko Ueta

1980-06-01

Full Text Available Foram registradas em Piquete, no vale do rio Paraíba do Sul (SP, Brasil, taxas de 1,22% e 0,14% de infecção natural em Lymnaea columella, por Fasciola hepatica. Em um único exemplar de Lymnaea columella dentre os 1.052 examinados, foram observadas rédias com xifidiocercárias, rédias com cercárias de Fasciola hepatica e metacercárias de Echinostomatidae.Infection rates of 1.22% and 0.14% were obtained in Lymnaea columella snails naturally infected by Fasciola hepatica. Samples of the snails were collected in Piquete, a municipality of Paraíba do Sul, a river valley area in the State of S. Paulo. Also observed was one of the 1052 specimen of the Lymnaea columella rediae which had xiphidiocercariae and rediae with Fasciola hepatica cercariae and metacercariae of Echinostomatidae.

Discovery of Maritrema jebuensis n. sp. (Digenea: Microphallidae) from the Asian Shore Crab, Hemigrapsus sanguineus, in Korea.

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Chung, Ok-Sik; Lee, Hye-Jung; Sohn, Woon-Mok; Lee, Seung-Ha; Park, Il-Yong; Oh, Sang-Ah; Chai, Jong-Yil; Seo, Min

2010-12-01

Maritrema spp. (Digenea: Microphallidae) are parasites of birds, but have not been found in the Republic of Korea. In this study, metacercariae of Maritrema sp. were discovered in the Asian shore crab, Hemigrapsus sanguineus, caught in the mud-flats of Jebu-do, Hwasung-gun, Gyeonggi-do, and the adult flukes were confirmed by experimental infection into mice. Based on the symmetric ribbon-like vitellarium, adult flukes of Maritrema sp. were identified, but did not belong to previously described species in terms of the following morphologic characteristics: ceca reaching to the lateral wall at the anterior border of the ovary; ventral sucker larger than oral sucker; a prominent metraterm; and vitellarium forming a complete ring. Hence, we named this microphallid M. jebuensis n. sp. after the island where the second intermediate hosts were collected. From this study, it has been shown that Maritrema sp. is distributed in Korea and transmitted by the Asian shore crab, H. sanguineus.

A case of a facultative life-cycle diversification in the fluke Pleurogenoides sp. (Lecithodendriidae, Plagiorchiida).

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Hassl, Andreas R

2010-10-01

Numerous specimens of the native, intestinal digenean fluke Pleurogenoides sp. (Lecithodendriidae, Plagiorchiida), a genus known for the simultaneous co-existence of genuine adults and progenetic, adult-like metacercaria, were found by chance parasitizing in the oesophagus of a recently imported, tropical Bristly Bush Viper (Atheris hispida). The snake had before been force-fed with native water frogs, the assumed definitive host of these flukes. Hence water frogs act as the second intermediate host or as a paratenic host for Pleurogenoides flukes, as they must house progenetic fluke larvae, which develop to genuine adults when transmitted to an appropriate consecutive host, the ancestral definitive host, a reptile. The European Pleurogenoides fluke species seem to display a facultative life-cycle diversification, they can adjust their life-history strategy according to their immediate transmission opportunities. This phenotypic plasticity allows the parasite to respond quickly to any changes in the abundance of a host; usually this biological oddity results in a life-cycle truncation by the elimination of the definitive host.

Melanoides tuberculata (Mollusca: Thiaridae as an intermediate host of Centrocestus formosanus (Trematoda: Heterophyidae in Brazil Melanoides tuberculata (Mollusca: Thiaridae como hospedeiro intermediário de Centrocestus formosanus (Trematoda: Heterophyidae no Brasil

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Hudson Alves Pinto

2010-08-01

Full Text Available Pleurolophocercous cercariae emerged from naturally infected Melanoides tuberculata from Minas Gerais State, Brazil, were used to perform experimental infection of laboratory-reared Poecilia reticulata. Mature metacercariae were obtained from the gills of fishes and force-fed to Mus musculus. The adult parasites which recovered from small intestines of mice were identified as Centrocestus formosanus. This is the first report of M. tuberculata as intermediate host of this heterophyid in Brazil.Cercárias do tipo pleurolofocerca emergidas de Melanoides tuberculata naturalmente infectados coletados na represa da Pampulha, Belo Horizonte, Minas Gerais, Brasil, foram utilizadas para a infecção experimental de Poecilia reticulata criados em laboratório. Metacercárias maduras foram obtidas nas brânquias dos peixes e administradas por via oral a Mus musculus. Parasitos adultos recuperados no intestino delgado dos camundongos foram identificados como Centrocestus formosanus. Este é o primeiro relato de M. tuberculata como hospedeiro intermediário deste parasito no Brasil.

A new species of Ascocotyle (Trematoda: Heterophyidae) from the South American sea lion, Otaria flavescens, off Patagonia, Argentina.

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Hernández-Orts, Jesús Servando; Montero, Francisco Esteban; Crespo, Enrique Alberto; García, Néstor Aníbal; Raga, Juan Antonio; Aznar, Francisco Javier

2012-08-01

We describe a new heterophyid species, Ascocotyle (Ascocotyle) patagoniensis n. sp., based on specimens collected from the intestines of the South American sea lion Otaria flavescens from Patagonia (Argentina). Ascocotyle (A.) patagoniensis n. sp. is distinguished from the other species of the subgenus by the number of circumoral spines, which are arranged in 2 rows of 18 to 23. The new species also differs from the other species in having a gonotyl without papillae. The specimens exhibited the widest seminal receptacle described for a species of this subgenus. Species of the subgenus Ascocotyle usually infect fish-eating birds or mammals in freshwater or brackish habitats. Ascocotyle (A.) patagoniensis n. sp. is the first species of the subgenus described from a marine mammal. However, no metacercariae of Ascocotyle spp. were found in 542 marine teleosts from 20 species collected in the same locality. The life cycle of the marine species from the Ascocotyle -complex infecting pinnipeds remains elusive.

Digenea, Nematoda, Cestoda, and Acanthocephala, parasites in Potamotrygonidae (Chondrichthyes from the upper Paraná River floodplain, states of Paraná and Mato Grosso do Sul, Brazil.

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Pavanelli, G. C.

2008-01-01

Full Text Available The present paper represents the first study on the endoparasitic fauna of Potamotrygon falkneri and P.motoro in the upper Paraná River floodplain. Fishes were collected by fishing rod and gillnetting in different stations ofthe floodplain, from March, 2005 to September, 2006. Parasites were sampled, fixed and preserved according tospecialized literature. About half of the analyzed fish were parasitized by at least one of the following species ofendoparasites: Clinostomum complanatum, Genarchella sp. and Tylodelphys sp. (metacercaria (Digenea;Acanthobothrium regoi, Rhinebothrium paratrygoni, Paroncomegas araya and Potamotrygonocestus travassosi(Cestoidea; Brevimulticaecum sp. (larva, Cucullanus sp., Echinocephalus sp. and Spinitectus sp. (Nematoda; andQuadrigyrus machadoi (Acanthocephala. Some species were already registered in Chondrichthyes and others werepreviously recorded in Osteichthyes from the study area. The study listed ten new records of parasites in the host P.falkneri, one new record in the host P. motoro and five new records in the locality upper Paraná River.

Application of radioisotopes in the diagnosis of fascioliasis. Part of a coordinated programme on isotopes and radiation in animal parasitology and immunology

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Tomanek, J.

1977-02-01

Fascioliasis is an important parasitic disease of grazing cattle and sheep. The experimental approach included the production of metacercariae of Fasciola hepatica from Lymnae tomentosa, infection of experimental animals, separation of fractions from whole Fasciola hepatica antigen by gel filtration on Sephadex G-100 and G-200, labelling of antigens with 125 I and examination of experimentally infected sheep and other animals for antibodies by modified radioactive single radial diffusion, indirect radioimmunodiffusion and radioactive antigen microprecipitin assay by means of direct radioimmunodiffusion and immunodiffusion using unlabelled antigen. Sufficient quantities of fluke for the production of antigen were obtained only from sheep. Infected rabbits, guinea pigs and rats yielded relatively few flukes. Labelling of the antigenic fraction can be regarded as fairly successful but the specific radioactivity of the fractions needs to be increased. Results from antibodies in infected animals by indirect radioimmunodiffusion and modified radioactive single diffusion proved not fully specific. Radioactive antigen microprecipitin assay would appear to differentiate between negative and positive sera

The immunological characteristics and probiotic function of recombinant Bacillus subtilis spore expressing Clonorchis sinensis cysteine protease.

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Tang, Zeli; Shang, Mei; Chen, Tingjin; Ren, Pengli; Sun, Hengchang; Qu, Hongling; Lin, Zhipeng; Zhou, Lina; Yu, Jinyun; Jiang, Hongye; Zhou, Xinyi; Li, Xuerong; Huang, Yan; Xu, Jin; Yu, Xinbing

2016-12-19

Clonorchiasis, a food-borne zoonosis, is caused by Clonorchis sinensis. The intestinal tract and bile ducts are crucial places for C. sinensis metacercariae to develop into adult worms. The endospore of Bacillus subtilis is an ideal oral immunization vehicle for delivery of heterologous antigens to intestine. Cysteine protease of C. sinensis (CsCP) is an endogenous key component in the excystment of metacercariae and other physiological or pathological processes. We constructed a fusion gene of CotC (a coat protein)-CsCP and obtained B. subtilis spores with recombinant plasmid of pEB03-CotC-CsCP (B.s-CotC-CsCP). CotC-CsCP expressed on spores' surface was detected by Western blotting and immunofluorescence. Immunological characteristics of recombinant spore coat protein were evaluated in a mouse model. The levels of CsCP-specific antibodies were detected by ELISA. Effects of recombinant spores on mouse intestine were evaluated by histological staining. The activities of biochemical enzymes in serum were assayed by microplate. Liver sections of infected mice were evaluated by Ishak score after Masson's trichrome. The B.s-CotC-CsCP spores displayed CsCP on their coat. Specific IgG and isotypes were significantly induced by coat proteins of B.s-CotC-CsCP spores after subcutaneous immunization. IgA levels in intestinal mucus and bile of B.s-CotC-CsCP orally treated mice significantly increased. Additionally, more IgA-secreting cells were observed in enteraden and lamina propria regions of the mouse jejunum, and an increased amount of acidic mucins in intestines were also observed. There were no significant differences in enzyme levels of serum among groups. No inflammatory injury was observed in the intestinal tissues of each group. The degree of liver fibrosis was significantly reduced after oral immunization with B.s-CotC-CsCP spores. Bacillus subtilis spores maintained the original excellent immunogenicity of CsCP expressed on their surface. Both local and systemic

Two-year intervention trial to control of fish-borne zoonotic trematodes in giant gourami (Osphronemus goramy) and striped catfish (Pangasianodon hypophthalmus) in nursery ponds in the Mekong Delta, Vietnam.

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Madsen, H; Thien, P C; Nga, H T N; Clausen, J H; Dalsgaard, A; Murrell, K D

2015-12-01

Fish-borne zoonotic trematode parasites (FZT) pose a food safety and public health problem in Vietnam. The transmission cycle is complex as domestic animals, especially dogs, cats, fish-eating birds and pigs together with humans serve as reservoir hosts and contribute to FZT egg contamination of aquaculture ponds and the environment. This intervention trial was conducted to determine the effectiveness of various on-farm interventions, including reduction in FZT egg contamination through treatment of infected people and domestic animals, reduction in snail density through mud removal from aquaculture ponds prior to fish stocking, and various other measures in reducing FZT infection in juvenile striped catfish (Pangasianodon hypophthalmus) and giant gourami (Osphronemus goramy). Interventions were implemented on 5 farms for each fish species during production cycles in 2009 and 2010 while 5 similar farms for each species served as control. For both fish species, both prevalence and intensity of infection did not differ significantly between intervention and non-intervention farms prior to the interventions. The interventions significantly reduced both prevalence and intensity of FZT infection in the juvenile fish compared to control ponds. For giant gourami, odds of infection in intervention ponds was 0.13 (95% CL: 0.09-0.20; p<0.001) of that in non-intervention ponds after the 2009 trial and 0.07 (0.03-0.14; p<0.001) after the 2010 trial. For striped catfish, these figures were 0.17 (0.08-0.35; p<0.001) after the 2009 trial while after the 2010 trial all ponds with interventions were free from infection. Metacercariae intensity (no. of metacercariae/fish) in giant gourami from intervention ponds was 0.16 (0.11-0.23; p<0.001) of that in fish from non-intervention ponds after the 2009 trial and 0.07 (0.04-0.15; p<0.001) after the 2010 trial; for striped catfish these figures were 0.18 (0.09-0.36; p<0.001) and 0.00 (confidence limits not estimated), respectively. The

Seroprevalence of Human Fasciolosis in Pirabad, Lorestan Province, Western Iran

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Farnaz KHEIRANDISH

2016-03-01

Full Text Available Background: The purpose of this study was to perform seroepidemiological investigation for determining the status of human fasciolosis in Pirabad Village, Lorestan Province, western Iran. Methods: Blood samples were taken from residents of the village including 801 individuals. Sera were separated and stored at -20°C until used. The samples were analyzed using ELISA. Results: Anti-Fasciola antibodies were detected in 6 (0.7% individuals. Difference between age, sex and drinking or swimming in the surface water with seropositivity to fasciolosis was not significant. Out of 7 shepherds, 1 (14.3% was seropositive. Due to the small number of shepherds, comprehensive statistical inference in this regard cannot be done. Significant difference was detected between seropositivity to fasciolosis and consuming local freshwater vegetables during the last 6 months (P=0.001.Conclusion: Metacercariae carrying local freshwater plants might be the main source of contamination because consumption of these kinds of vegetables was confirmed by all participants. Awareness of local communities regarding the danger of freshwater plant consumption, through health education programs, will decrease the risk of infection.

Discovery of Paragonimus westermani in Vietnam and its molecular phylogenetic status in P. westermani complex.

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Doanh, Pham Ngoc; Shinohara, Akio; Horii, Yoichiro; Habe, Shigehisa; Nawa, Yukifumi

2009-04-01

Paragonimus westermani is the most well-known species among the genus Paragonimus. It is widely distributed in Asia with considerable genetic diversity to form P. westermani species complex. While P. westermani distributed in Japan, Korea, China, and Taiwan are genetically homogeneous to form the East Asia group, those found in other geographic areas are heterogeneous and would be divided into several groups. Recent discoveries of P. westermani in India and Sri Lanka highlighted new insights on molecular phylogenetic relationship of geographic isolates of this species complex. Since Vietnam is located at the east end of Southeast Asia, the intermediate position between South and East Asia, it is of interest to see whether P. westermani is distributed in this country. Here, we report that P. westermani metacercariae were found in mountainous crabs, Potamiscus sp., collected in Quangtri province in the central Vietnam. Adult worms were successfully obtained by experimental infection in cats. Molecular phylogenetic analyses revealed that P. westermani of Vietnamese isolates have high similarities with those of East Asia group.

Community of protozoans and metazoans parasitizing Auchenipterus nuchalis (Auchenipteridae, a catfish from the Brazilian Amazon

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Marcos Tavares Dias

2017-05-01

Full Text Available This paper describes the first study on parasite diversity in Auchenipterus nuchalis Spix & Agassiz, 1829 (Auchenipteridae. In 31 fish caught in a tributary of the Amazon River, 10,708 parasites were collected, such as Ichthyophthirius multifiliis, Piscinoodinium pilullare, Cosmetocleithrum striatuli, metacercariae of Posthodiplostomum sp, and larvae and adults of Procamallanus (Spirocamallanus inopinatus. These parasite species showed aggregated dispersion, except for C. striatuli, which had a uniform dispersion. The component community of parasites showed a low Brillouin diversity (0.67 ± 0.27, low species richness (3.5 ± 0.8 and low evenness (0.43 ± 0.17, and it was characterized by the presence of species with high prevalence and abundance. Protozoan species were the prevalent parasites, which may be a consequence of the host’s mode of life, while the low presence of endoparasites in A. nuchalis suggests that this fish may occupy an intermediate trophic level in the food web. These data represent the first record of such parasite species for this host.

Protection against heterologous infection by using cross antigenicity between schistosoma mansoni and fasciola hepatica

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Moawad, M.A.; Amin, M.M.

2004-01-01

Fasciola hepatica is the causative agent of fasciolosis in many areas in America, Europe, Africa, Asia and Australia. There is an urgent need for improved methods to control the parasite's transmission. The present study is parasitological, immunological (interleukine-1β and interleukine-6) and histopathological investigations on the immunizing effect of cross antigenicity between S. mansoni and F. hepatica against schistosomiasis and fasciolosis in mice. Parasitological study showed that vaccination with irradiated cercariae of S. mansoni or vaccination by F. hepatica whole worm extract (FhWWE) before challenged with encysted metacercariae of F. hepatica or cercariae of S. mansoni played a significant control on the parasitic infection manifested by a remarkable reduction in the means of worm count. Assessment of IL-1β and IL-6 in sera of the experimental groups showed that there are cross reactivity between fasciola / schistosoma and its relation to cross protection. Histopathological examination of vaccinated mice livers showed protection against parasite maturation and liver damage after challenged, as compared to mice infected only without vaccination

[Epidemiologic and clinical study of paragonimosis in Cameroon. Results of niclofolan treatment].

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Ripert, C; Carrie, J; Ambroise-Thomas, P; Baecher, R; Kum, N P; Same-Ekobo, A

1981-01-01

Paragonimiasis does not occur only in the South West Province of Cameroon. Four foci exist in the country. They are the well known focus in the Mount Kupe area, the Mbam focus, the Nyong focus and the Ntem focus. All of them are located in the rainforest within the distribution area of Potadoma. This disease, characterized by the presence of eggs in sputum and feces, has often been confused with tuberculosis. In the villages where paragonimiasis is diagnosed by means of parasitological techniques circulating antibodies, revealed with a P. westermani antigen (ELISA), are often found in blood specimens taken from inhabitants. Teen agers suffer the most from the disease and females more often than males. Women and children are traditionally concerned with crabs fishing and they eat them after partly raw. Crabs of the genus Sudanautes contain the infective metacercariae. Paragonimiasis is enzootic and the civet cat V. civetta seems to be the main natural definitive host in Cameroon. Niclofolan given orally in a single dose of 2 mg/kg body weight showed a 100% cure rate. Side reactions are mild and transient.

Heterophysiasis, an intestinal fluke infection of man and vertebrates transmitted by euryhaline gastropods and fish

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Taraschewski, H.

1984-03-01

Heterophyes heterophyes, agent of human heterophyiasis in the Near East, is transmitted in marine lagoons and saline inland waters, where the euryhaline intermediate hosts are abundant. In Egypt, mullets, the predominant second intermediate hosts, are customarily consumed raw; thus man becomes infected easily. Symptoms of human infections are usually considered mild. Mullets do not seem to be affected by the metacercariae encysted in the muscles, whereas the growth of the snail host Pirenella conica was found to be enhanced due to the infestation by the trematodes. In laboratory experiments, the flukes were found to be well developed in dogs, foxes and cats, but failed to reach sexual maturity in several other potentially piscivorous mammals and birds. In nature, dogs probably serve as the major reservoir hosts. Heterophyiasis is most prevalent in the Nile Delta, a huge brackish water area which is densely populated by humans and, consequently, also by dogs and cats. In the Far East, besides Heterophyes nocens, several other heterophysids with marine or fresh-water life-cycles are known to infect humans.

Observations on the pathogenesis of anaemia in fascioliasis and on immunoglobulin metabolism

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Ogunrinade, A.F.; Makinde, M.O.

1988-01-01

Proline, an excretory/secretory product of liver flukes, was investigated as a possible mediator of anaemia through haemolysis or dyshaemopoiesis in fluke infected animals. Infusion of 700 μmol proline per rabbit per day for 14 days did not significantly alter the halflife (t 1/2 ) of 51 Cr labelled erythrocytes nor did it interfere with plasma clearance and utilization of 59 Fe in rabbits. Consequently, it was concluded that proline did not interfere with haemoglobin synthesis or induced haemolysis in rabbits at test dose level. The metabolism of radiolabelled IgG was also investigated in calves infected with Fasciola gigantica. In two calves given 1,000 metacercariae of F. gigantica ten weeks earlier, the mean t 1/2 of 125 I labelled IgG was 74 h, compared with 100 h in uninfected controls. The faecal clearance of 125 I labelled IgG was correspondingly higher among infected calves, indicating that serum IgG was being lost into the gut in excessive quantities, probably via the bile. (author). 20 refs, 4 tabs

Experimental murine fascioliasis derives early immune suppression with increased levels of TGF-β and IL-4.

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Chung, Joon-Yong; Bae, Young-An; Yun, Doo-Hee; Yang, Hyun-Jong; Kong, Yoon

2012-12-01

In fascioliasis, T-helper 2 (Th2) responses predominate, while little is known regarding early immune phenomenon. We herein analyzed early immunophenotype changes of BALB/c, C57BL/6, and C3H/He mice experimentally infected with 5 Fasciola hepatica metacercariae. A remarkable expansion of CD19(+) B cells was observed as early as week 1 post-infection while CD4(+)/CD8(+) T cells were down-regulated. Accumulation of Mac1(+) cells with time after infection correlated well with splenomegaly of all mice strains tested. The expression of tumor necrosis factor (TNF)-α mRNA in splenocytes significantly decreased while that of IL-4 up-regulated. IL-1β expression was down-modulated in BALB/c and C57BL/6 mice, but not in C3H/He. Serum levels of transforming growth factor (TGF)-β were considerably elevated in all mice during 3 weeks of infection period. These collective results suggest that experimental murine fascioliasis might derive immune suppression with elevated levels of TGF-β and IL-4 during the early stages of infection.

RNAi-mediated silencing of enolase confirms its biological importance in Clonorchis sinensis.

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Wang, Xiaoyun; Chen, Wenjun; Tian, Yanli; Huang, Yan; Li, Xuerong; Yu, Xinbing

2014-04-01

Clonorchis sinensis (C. sinensis) infection is still a common public health problem in freshwater fish consumption areas in Asian countries. More molecular evidence are required to speed up the prevention strategies to control this kind of infectious disease. In the present study, to confirm the biological importance of Csenolase followed by our previous observations of the key metabolic enzyme, we explored the RNA silence effect of the Csenolase-derived RNA interference (RNAi) in C. sinensis. The extramembranous region aa105-226 was selected as the target sequence of RNA silence. Csenolase-derived double strand RNA (dsRNA-Csenolase, 366 bp) was synthetized and delivered into C. sinensis by soaking approach. The penetration of dsRNA into adult worms and metacercariae was tracked using fluorescently labeled RNA. Western blotting and qRT-PCR experiments were performed to determine dsRNA-Csenolase-silencing effect. Our results showed that, after incubating for 120 h, dsRNA-Csenolase could effectively target and downregulate the expression of Csenolase in both adult worms (P sinensis adult worms (P sinensis, allowing further applications in identifying functional genes in C. sinensis.

Molecular analyses reveal high species diversity of trematodes in a sub-Arctic lake

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Soldánová, Miroslava; Georgieva, Simona; Roháčováa, Jana; Knudsen, Rune; Kuhn, Jesper A.; Henriksen, Eirik H.; Siwertsson, Anna; Shaw, Jenny C.; Kuris, Armand M.; Amundsen, Per-Arne; Scholz, Tomáš; Lafferty, Kevin D.; Kostadinova, Aneta

2017-01-01

To identify trematode diversity and life-cycles in the sub-Arctic Lake Takvatn, Norway, we characterised 120 trematode isolates from mollusc first intermediate hosts, metacercariae from second intermediate host fishes and invertebrates, and adults from fish and invertebrate definitive hosts, using molecular techniques. Phylogenies based on nuclear and/or mtDNA revealed high species richness (24 species or species-level genetic lineages), and uncovered trematode diversity (16 putative new species) from five families typical in lake ecosystems (Allocreadiidae, Diplostomidae, Plagiorchiidae, Schistosomatidae and Strigeidae). Sampling potential invertebrate hosts allowed matching of sequence data for different stages, thus achieving molecular elucidation of trematode life-cycles and exploration of host-parasite interactions. Phylogenetic analyses also helped identify three major mollusc intermediate hosts (Radix balthica, Pisidium casertanum and Sphaerium sp.) in the lake. Our findings increase the known trematode diversity at the sub-Arctic Lake Takvatn, showing that digenean diversity is high in this otherwise depauperate sub-Arctic freshwater ecosystem, and indicating that sub-Arctic and Arctic ecosystems may be characterised by unique trematode assemblages.

Endohelminth parasites of the freshwater fish Zoogoneticus purhepechus (Cyprinodontiformes: Goodeidae from two springs in the Lower Lerma River, Mexico Endohelmintos parásitos del pez dulceacuícola Zoogoneticus purhepechus (Cyprinodontiformes: Goodeidae en dos manantiales de la cuenca del río Lerma bajo, México

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Andrés Martínez-Aquino

2011-12-01

Full Text Available In order to establish the helminthological record of the viviparous fish species Zoogoneticus purhepechus, 72 individuals were collected from 2 localities, La Luz spring (n= 45 and Los Negritos spring (n= 27, both in the lower Lerma River, in Michoacán state, Mexico. Twelve helminth taxa were recovered, 5 adults (the digeneans Margotrema bravoae and Phyllodistomum sp., the cestode Bothriocephalus acheilognathi, the nematode Rhabdochona lichtenfelsi and the acanthocephalan Pomphorhynchus cf. bulbocolli, and 7 larvae (the metacercariae of Clinostomum complanatum, the cysticercoid of Cyclophyllidea, the nematodes Rhabdochona sp., Eustrongylides sp., Contracaecum sp. and Spiroxys sp., and the cysthacanth of Polymorphus brevis. Of these, R. lichtenfelsi was the most prevalent and abundant species at La Luz spring with 15.6% and 0.33 individuals per analyzed host. The remaining species were relatively more rare and infrequent. The helminth parasite community of Z. purhepechus at Los Negritos spring was remarkably poor and abundance was very low. The omnivorous feeding habits, the position of the host species in the food web, and the environmental characteristics of each locality are suggested as the main factors determining the helminth parasite communities in this freshwater fish.Se examinaron 72 individuos del pez vivíparo Zoogoneticus purhepechus para establecer el registro helmintológico de la especie. Los huéspedes se recolectaron de los manantiales La Luz (n= 45 y Los Negritos (n= 27, ubicados en la porción baja del río Lerma, en el estado de Michoacán, México. El registro helmintológico consta de 12 especies, incluyendo como adultos los digéneos Margotrema bravoae y Phyllodistomum sp., el céstodo Bothriocephalus acheilognathi, el nemátodo Rhabdochona lichtenfelsi, y el acantocéfalo Pomphorhynchus cf. bulbocolli. Además, como estadios larvarios, se encontraron las metacercarias de Clinostomum complanatum, el cisticercoide de

CONTRIBUTION A L'ETUDE DE LA DISTOMATOSE HEPATIQUE EN IRAN: RAPPORT DE DEUX CAS DE DISTOMATOSE OPERES COMME CAS DE LITHIASE BILIAIRE

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SEYED FARSHI DJALAL

1985-05-01

Full Text Available The a ut ho rs e xpl ain and review " very br iefly the Dis t omatosis i n human and animals . They f ound out that t h i s par a s ite is p resent i n Ovine i n a wide r ange i n I ran . This zoonosis and human contami na t ion from a ni ma ls (Fixed or freely f l oa t i ng metacercaria e l ma i n l y s heep , s houl d be mor e pr e valent in Iran and neighbouring CQunt r ies , with a hyg ienic and a s ocia economic i mportanc e, where people are a c c us t umed to eat crude vegetables . The authors observed t .....o cases of Human Fasciolosis due t o F . hepatica , describing symptomes, confused with common bile duct s tan . The y r ecomme nd an i nt e nse and treatment of main s ucepti ble animals regul a r l y and e a r l y det e ction and tre atmen t of infes tatio n in human.

Endohelminths in Cichla piquiti(Perciformes, Cichlidae from the Paraná River, São Paulo State, Brazil

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Lidiane Franceschini

Full Text Available Fifty specimens of Cichla piquiti were collected from the Paraná River downstream of the Ilha Solteira Hydroelectric Power Station in Brazil and surveyed for endohelminth parasites. All fish were parasitised by at least one helminth species (overall prevalence [P] = 100%. Eight parasite taxa were present: the nematode Procamallanus(Procamallanus peraccuratus in the intestines; third-stage larvae of the anisakids Contracaecumsp. and Hysterothylacium sp. in the visceral cavity, mesentery and serosa of the stomach and intestines and on the liver and spleen; the trematodes Austrodiplostomum compactum in the eye (metacercariae and Genarchella genarchella in the stomach; and the cestodes Proteocephalus macrophallus, Proteocephalus microscopicus, and Sciadocephalus megalodiscus in the intestines. Hysterothylaciumsp. larvae (P = 86% and P. microscopicus (P = 74% were the most prevalent parasites. Anisakids were more prevalent and abundant in the dry season. A negative correlation between the abundances of Hysterothylacium sp. and P. microscopicuswas observed, suggesting a competitive/antagonistic relationship between these parasites. Cichla piquiti represents a new host for four parasite species. These new records significantly increase the list of parasites of C. piquiti, contributing to the knowledge of the host-parasite relationship and the geographical distribution of these helminths.

Parasites of the mangrove mussel Mytella guyanensis (Bivalvia: Mytilidae) in Camamu Bay, Bahia, Brazil.

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Ceuta, L O; Boehs, G

2012-08-01

This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E). The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs), Nematopsis sp. (Apicomplexa), and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05) of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.

The susceptibility differences of buffalo and Ongole calves against trickle infection with Fasciola gigantica

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Ening Wiedosari

1998-03-01

Full Text Available A pen trial was carried out in order to determine the susceptibility differences of a trickle infection with Fasciola gigantica in buffalo and Ongole calves. Treated animals were infected orally with 15 metacercariae of F. gigantica twice weekly for 32 weeks and were slaughtered at 36 weeks. The results showed that buffalo calves had significantly lower fluke burdens than Ongole calves (P<0 .01 . All of the infected Ongole calves had fasciola eggs in their faeces 18 weeks after the commencement of infection. In contrast, eggs were detected only in 3 out of 7 infected buffalo calves at week 20, in 4 at week 28 and in 6 at week 30. Faeces of the seventh buffalo remained free of eggs until week 36. Rates of growth were reduced by 25%and 10,3% in infected Ongole and buffalo calves respectively . Circulating blood eosinophilia ofboth hosts, ahallmark of helminth infections, increased following infection, but values in buffaloes was greater than Ongole calves mainly in week 4 and 8 after infection (P<0.01. These results might be concluded that the susceptibility of buffalo calves to trickle infection with F. gigantica was lower compared to Ongole calves.

A molecular phylogenetic appraisal of the acanthostomines Acanthostomum and Timoniella and their position within Cryptogonimidae (Trematoda: Opisthorchioidea).

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Martínez-Aquino, Andrés; Vidal-Martínez, Victor M; Aguirre-Macedo, M Leopoldina

2017-01-01

The phylogenetic position of three taxa from two trematode genera, belonging to the subfamily Acanthostominae (Opisthorchioidea: Cryptogonimidae), were analysed using partial 28S ribosomal DNA (Domains 1-2) and internal transcribed spacers (ITS1-5.8S-ITS2). Bayesian inference and Maximum likelihood analyses of combined 28S rDNA and ITS1 + 5.8S + ITS2 sequences indicated the monophyly of the genus Acanthostomum ( A. cf. americanum and A. burminis ) and paraphyly of the Acanthostominae . These phylogenetic relationships were consistent in analyses of 28S alone and concatenated 28S + ITS1 + 5.8S + ITS2 sequences analyses. Based on molecular phylogenetic analyses, the subfamily Acanthostominae is therefore a paraphyletic taxon, in contrast with previous classifications based on morphological data. Phylogenetic patterns of host specificity inferred from adult stages of other cryptogonimid taxa are also well supported. However, analyses using additional genera and species are necessary to support the phylogenetic inferences from this study. Our molecular phylogenetic reconstruction linked two larval stages of A. cf. americanum cercariae and metacercariae. Here, we present the evolutionary and ecological implications of parasitic infections in freshwater and brackish environments.

A molecular phylogenetic appraisal of the acanthostomines Acanthostomum and Timoniella and their position within Cryptogonimidae (Trematoda: Opisthorchioidea

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Andrés Martínez-Aquino

2017-12-01

Full Text Available The phylogenetic position of three taxa from two trematode genera, belonging to the subfamily Acanthostominae (Opisthorchioidea: Cryptogonimidae, were analysed using partial 28S ribosomal DNA (Domains 1–2 and internal transcribed spacers (ITS1–5.8S–ITS2. Bayesian inference and Maximum likelihood analyses of combined 28S rDNA and ITS1 + 5.8S + ITS2 sequences indicated the monophyly of the genus Acanthostomum (A. cf. americanum and A. burminis and paraphyly of the Acanthostominae. These phylogenetic relationships were consistent in analyses of 28S alone and concatenated 28S + ITS1 + 5.8S + ITS2 sequences analyses. Based on molecular phylogenetic analyses, the subfamily Acanthostominae is therefore a paraphyletic taxon, in contrast with previous classifications based on morphological data. Phylogenetic patterns of host specificity inferred from adult stages of other cryptogonimid taxa are also well supported. However, analyses using additional genera and species are necessary to support the phylogenetic inferences from this study. Our molecular phylogenetic reconstruction linked two larval stages of A. cf. americanum cercariae and metacercariae. Here, we present the evolutionary and ecological implications of parasitic infections in freshwater and brackish environments.

Zoonotic ectopic fascioliasis: review and discussion.

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Rashed, Amr A; Khalil, Hazem H M; Morsy, Ayman T A

2010-12-01

Ectopic fascioliasis (EF) has direct and indirect effects on both humans and animals. The phenomenon of EF was individual cases in the period from 1950 up to the end of last century. From the period of 2000 up to 2006, plenty of researches were on EF in the developed and undeveloped countries. Nineteen EF cases infected with the immature and few with the mature worms were 13 females and 6 males. Three cases of lymphatic, pleural and breast fascioliasis reached the adults and laid their eggs in a lymph node in the cervical region pleural cavity and breast tissues. Until recent, knowledge about the ectopic fascioliasis pathway is little. Fasciola hepatica was the commonest species in most cases. The effect of fascioliasis might be direct to liver as ectopic foci or indirect on other organs due to the metabolites and secretory excretory products. All ages and both sexes were EF infected. Watercress topped the list of water plants born encysted metacercariae followed by lettuce, mint, and alfalfa. Nearly 24 million Egyptians at risk and about 800,000 were infected. On the global scale, about 180 million are at risk of infection.

The occurence of black spot disease in Astyanax aff. fasciatus(characiformes: characidae in the Guaíba Lake basin, RS, Brazil

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F Flores-Lopes

Full Text Available Black spot disease is common in freshwater fish and is usually caused by the metacercaria stage of digenetic trematodes, normally from the Diplostomidae family. The present study evaluated the prevalence and intensity of this disease in Astyanax aff. fasciatus(Teleostei: Characiformes in the Guaíba Lake basin (RS, Brazil, including body parts assessment and the points of sampling with higher occurrence of black spots. Fish samples were taken seasonally from December 2002 until October 2004. The samples were collected with the use of a seine net at eleven points. The specimens were fixed in 10% formalin and stored in 70% ethanol. Black spot disease showed a low frequency in the Guaíba lake basin (2.07% and no specificity to the species Astyanax aff. fasciatus was observed. A high prevalence among the individuals and high intensity of infection levels was found in the ventral and dorsal regions in relation to other body parts (e.g., pectoral, pelvic and anal regions. Among the sampling points studied, we observed a higher prevalence on samples collected at points Gasômetro, Saco da Alemoa and Sinos, located in open areas with less occurrence of mollusks.

Parasites of the mangrove mussel Mytella guyanensis (Bivalvia: Mytilidae in Camamu Bay, Bahia, Brazil

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LO. Ceuta

Full Text Available This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E. The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs, Nematopsis sp. (Apicomplexa, and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05 of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.

Virus-like particles vaccine containing Clonorchis sinensis tegumental protein induces partial protection against Clonorchis sinensis infection.

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Lee, Dong-Hun; Kim, Ah-Ra; Lee, Su-Hwa; Quan, Fu-Shi

2017-12-29

Human clonorchiasis, caused by the infection of Clonorchis sinensis, is one of the major health problems in Southeast Asia. However, vaccine efficacy against C. sinensis infection remains largely unknown. In this study, for the first time, we generated virus-like particles (VLPs) vaccine containing the C. sinensis tegumental protein 22.3 kDa (CsTP 22.3) and the influenza matrix protein (M1) as a core protein, and investigated the vaccine efficacy in Sprague-Dawley rats. Intranasal immunization of VLPs vaccine induced C. sinensis-specific IgG, IgG2a and IgG2c in the sera and IgA responses in the feces and intestines. Notably, upon challenge infection with C. sinensis metacercariae, significantly lower adult worm loads (70.2%) were measured in the liver of rats immunized with VLPs, compared to those of naïve rats. Furthermore, VLPs immunization induced antibody secreting cells (ASC) responses and CD4+/CD8+ T cell responses in the spleen. Our results indicated that VLPs vaccine containing C. sinensis CsTP 22.3 kDa provided partial protection against C. sisnensis infection. Thus, VLPs could be a potential vaccine candidate against C. sinensis.

Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini using real-time PCR and high resolution melting analysis.

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Cai, Xian-Quan; Yu, Hai-Qiong; Li, Rong; Yue, Qiao-Yun; Liu, Guo-Hua; Bai, Jian-Shan; Deng, Yan; Qiu, De-Yi; Zhu, Xing-Quan

2014-01-01

Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

Rapid Detection and Differentiation of Clonorchis sinensis and Opisthorchis viverrini Using Real-Time PCR and High Resolution Melting Analysis

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Xian-Quan Cai

2014-01-01

Full Text Available Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

Molecular characterization of cathepsin B from Clonorchis sinensis excretory/secretory products and assessment of its potential for serodiagnosis of clonorchiasis

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Zhou Chenhui

2011-07-01

Full Text Available Abstract Background Cathepsin cysteine proteases play multiple roles in the life cycle of parasites such as food uptake, immune invasion and pathogenesis, making them valuable targets for diagnostic assays, vaccines and drugs. The purpose of this study was to identify a cathepsin B of Clonorchis sinensis (CsCB and to investigate its diagnostic value for human helminthiases. Results The predicted amino acid sequence of the cathepsin B of C. sinensis shared 63%, 52%, 50% identity with that of Schistosoma japonicum, Homo sapiens and Fasciola hepatica, respectively. Sequence encoding proenzyme of CsCB was overexpressed in Escherichia coli. Reverse transcription PCR experiments revealed that CsCB transcribed in both adult worm and metacercaria of C. sinensis. CsCB was identified as a C. sinensis excretory/secretory product by immunoblot assay, which was consistent with immunohistochemical localization showing that CsCB was especially expressed in the intestine of C. sinensis adults. Both ELISA and western blotting analysis showed recombinant CsCB could react with human sera from clonorchiasis and other helminthiases. Conclusions Our findings revealed that secreted CsCB may play an important role in the biology of C. sinensis and could be a diagnostic candidate for helminthiases.

Characterization of immunogenic Clonorchis sinensis protein fractions by gel fitration chromatography

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Duan Pham Ngoc

2015-04-01

Full Text Available Objective: To characterize immunogenic protein fraction of Clonorchis sinensis (C. sinensis by partial purification. Methods: A total of 30 hamsters were infected with 50 C. sinensis metacercariae, and then C. sinensis protein was purified by gel filtration chromatography. Indirect ELISA and immunoblot were used to detect the antibody in sera of hamsters infected with C. sinensis. Results: The gel filtration showed 2 peaks at high (fraction No. 10 to 14 and low (fraction No. 21 to 26 molecular weight proteins. Indirect ELISA showed that both antibodies of clonorchiasis and opisthorchiasis reacted strongly with early fractions (6 to 14 and the reaction was gradually reduced at middle and late fractions (15 to 50. Both antibodies showed different individual fraction of C. sinensis by immunoblot. It showed several protein bands that the 34 and 37 kDa were major proteins. The 53 kDa protein which was only found in the clonorchiasis reacted with fraction 20. Conclusions: The purified antigen of C. sinensis reacted similarly with both antibodies of clonorchiasis and opisthorchiasis where strong reaction was seen with early fractions. The C. sinensis protein fraction No. 20 may be useful for immunodiagnosis of clonorchiasis.

Distribution of freshwater snails in family-based VAC ponds and associated waterbodies with special reference to intermediate hosts of fish-borne zoonotic trematodes in Nam Dinh Province, Vietnam.

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Dung, Bui Thi; Madsen, Henry; The, Dang Tat

2010-10-01

Fish-borne zoonotic trematodes, such as Clonorchis sinensis, heterophyids and others, constitute a public health concern in parts of northern Vietnam and infections with these trematodes are often thought to be linked to fish culture. One common fish culture system is the integrated fish-livestock (VAC) ponds where individual households have 1 or more ponds. Fish fry, mainly of various carp species, produced in hatcheries, not necessarily local, are introduced into nursery ponds and after approximately 6 weeks, juvenile fishes are transferred to household ponds, referred to as grow-out ponds. Grow-out ponds are usually fertilized with organic debris, including animal excreta, to stimulate algal growth and subsequently fish growth. This paper describes the distribution of freshwater snails and occurrence of trematode infections in these in VAC ponds and associated habitats as part of a major study on risk factors of FZT infections in cultured fish in two communes, Nghia Lac and Nghia Phu, Nghia Hung District, Nam Dinh Province. The area is under intense rice cultivation with an extensive canal network supplying fields and also household VAC ponds. A total of 16 snail species was found and four were widely distributed i.e. Angulyagra polyzonata, Melanoides tuberculata, Bithynia fuchsiana and Pomacea insularum. Snail diversity and counts were higher in nursery ponds than in grow-out ponds. Species of the families Thiaridae and Viviparidae were more abundant than other species in VAC ponds while species of the Bithyniidae, Stenothyridae and Planorbidae dominated in rice fields and small canals. Trematode infections were found in eight snail species and among these M. tuberculata had the highest overall prevalence of infection (13.28%). No trematode infections were found in species of the Viviparidae and Ampullaridae except for metacercariae. Parapleurolophocercous and pleurolophocercous cercariae constituted the most common type of cercariae recovered, contributing 40

Optimization of Conditions for In Vitro Culture of the Microphallid Digenean Gynaecotyla adunca

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Jenna West

2014-01-01

Full Text Available In vitro cultivation of digeneans would aid the development of effective treatments and studies of the biology of the parasites. The goal of this study was to optimize culture conditions for the trematode, Gynaecotyla adunca. Metacercariae of the parasite from fiddler crabs, Uca pugnax, excysted in trypsin, were incubated overnight to permit fertilization, and were cultured in different conditions to find those that resulted in maximum worm longevity and egg production. When cultured in media lacking serum, worms lived longer in Hanks balanced salt solution and Dulbecco’s Modified Eagle medium/F-12 (DME/F-12 than in RPMI-1640 but produced the most eggs in DME/F-12. Worm longevity and egg production increased when worms were grown in DME/F-12 supplemented with 20% chicken, horse, or newborn calf serum but the greatest number of eggs was deposited in cultures containing horse or chicken serum. Horse serum was chosen over chicken serum due to the formation of a precipitate in chicken serum. The optimal concentration of horse serum with respect to egg production ranged from 5 to 20%. Infectivity of eggs deposited by worms in culture was tested by feeding eggs to mud snails, Ilyanassa obsoleta. None of these snails produced G. adunca cercariae.

Immunoexpression of intermediate filaments and morphological changes in the liver and bile duct of rats infected with Fasciola hepatica.

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Kolodziejczyk, L; Laszczyńska, M; Masiuk, M; Grabowska, M; Skrzydlewska, E

2015-01-01

We investigated the immunoexpression of the intermediate filament proteins, cytokeratin and desmin, and the morphological changes in the liver of rats during experimental fasciolosis at 4, 7 and 10 weeks post-infection. Rats were infected with 30 Fasciola hepatica metacercariae. Paraffin sections of the liver were stained using H & E, PAS and azan stains. Immunohistochemical reactions were performed using antibodies against cytokeratin and desmin. The experimental F. hepatica infection led to fibrosis and cirrhosis of the liver, and to inflammation of the common bile ducts. The expression of cytokeratin was increased in the epithelial cells of both the liver bile ductules at 4, 7 and 10 weeks post-infection and in the common bile ducts at 7 and 10 weeks post-infection compared to uninfected rats; expression in the common bile ducts was more intense. The myofibroblasts of the liver and smooth myocytes of the interlobular bile ducts and common bile ducts, showed a slight increase in desmin expression compared to the uninfected rats. The increased expression of cytokeratins in the hyperplastic rat common bile duct epithelium during the biliary phase of fasciolosis at 7 and 10 weeks post-infection may be explained by mechanical irritation by the parasite and an inflammatory reaction in the bile duct epithelium and in periductal fibrous tissue.

Habitat of in vivo transformation influences the levels of free radical scavengers in Clinostomum complanatum: implications for free radical scavenger based vaccines against trematode infections.

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Zafar, Atif; Rizvi, Asim; Ahmad, Irshad; Ahmad, Masood

2014-01-01

Since free radical scavengers of parasite origin like glutathione-S-transferase and superoxide dismutase are being explored as prospective vaccine targets, availability of these molecules within the parasite infecting different hosts as well as different sites of infection is of considerable importance. Using Clinostomum complanatum, as a model helminth parasite, we analysed the effects of habitat of in vivo transformation on free radical scavengers of this trematode parasite. Using three different animal models for in vivo transformation and markedly different sites of infection, progenetic metacercaria of C. complanatum were transformed to adult ovigerous worms. Whole worm homogenates were used to estimate the levels of lipid peroxidation, a marker of oxidative stress and free radical scavengers. Site of in vivo transformation was found to drastically affect the levels of free radical scavengers in this model trematode parasite. It was observed that oxygen availability at the site of infection probably influences levels of free radical scavengers in trematode parasites. This is the first report showing that habitat of in vivo transformation affects levels of free radical scavengers in trematode parasites. Since free radical scavengers are prospective vaccine targets and parasite infection at ectopic sites is common, we propose that infections at different sites, may respond differently to free radical scavenger based vaccines.

Microphallids in Gammarus insensibilis Stock, 1966 from a Black Sea lagoon: manipulation hypothesis going East?

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Kostadinova, A; Mavrodieva, R S

2005-09-01

Patterns of parasite site selection, variation in infection parameters and interspecific associations are examined in the light of new field data on larval microphallids in Gammarus insensibilis from a Black Sea lagoon. These patterns are discussed in relation to the predictions for the manipulative effect of Microphallus papillorobustus and its relationships with the other microphallid species based on studies on the French Mediterranean coast. Four species were recovered: Maritrema subdolum, Microphallus hoffmanni, M. papillorobustus and Levinseniella propinqua. The latter two were located in both corporal and cephalic segments, but the selection of brain appeared stronger for L. propinqua. M. subdolum was the first colonizer of amphipod population recruits, and unequivocally the dominant species in the lagoon. There was a significant positive relationship between the parasite load of all 4 species. Concurrent infections were exceedingly frequent, and no departures from random association were detected. We found no evidence that the cerebral metacercariae of M. papillorobustus consistently predict the parasite load of any of the other species in the system and identify sources for heterogeneity that may account for the differences between the Black Sea and the Mediterranean system: habitat heterogeneity, bird diversity and host-parasite systems used to infer relationships between microphallids.

Trematode diversity in freshwater fishes of the Globe II: 'New World'.

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Choudhury, Anindo; Aguirre-Macedo, M Leopoldina; Curran, Stephen S; de Núñez, Margarita Ostrowski; Overstreet, Robin M; de León, Gerardo Pérez-Ponce; Santos, Cláudia Portes

2016-03-01

We provide a summary overview of the diversity of trematode parasites in freshwater fishes of the 'New World', i.e. the Americas, with emphasis on adult forms. The trematode fauna of three regions, South America, Middle America, and USA and Canada (North America north of Mexico), are considered separately. In total, 462 trematode species have been reported as adults from the Americas. The proportion of host species examined for parasites varies widely across the Americas, from a high of 45% in the Mexican region of Middle America to less than 5% in South America. North and South America share no adult species, and one exclusively freshwater genus, Creptotrema Travassos, Artigas & Pereira, 1928 in the Allocreadiidae Looss, 1902 is the most widely distributed. Metacercariae of strigeiforms maturing in fish-eating birds (e.g. species of the Diplostomidae Poirier, 1886) are common and widely distributed. The review also highlights the paucity of known life-cycles. The foreseeable future of diversity studies belongs to integrative approaches and the application of molecular ecological methods. While opportunistic sampling will remain important in describing and cataloguing the trematode fauna, a better understanding of trematode diversity and biology will also depend on strategic sampling throughout the Americas.

An incidental case of biliary fascioliasis mimicking cholangiocellular carcinoma.

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Şenateş, Ebubekir; Doğan, Abdullah; Şenates, Banu Erkalma; Bodakçi, Erdal; Bekçibasi, Muhammet

2014-12-01

Fascioliasis is a zoonotic infestation caused by Fasciola hepatica that usually attacks mammals, such as goats, sheep and cattle. The parasites can infect humans via freshwater plants contaminated with encysted metacercariae. In the acute phase, which involves hepatic invasion, the disease may present with abdominal pain, mild fever and hepatomegaly. In the chronic phase, the parasites settle into the biliary tracts, and then cause cholangitis and cholestasis. Sometimes, the disease may mimic malignancies, creating a mass appearance. Endoscopic retrograde cholangiopancreatography (ERCP) is an important diagnostic and treatment method because it allows simultaneous diagnosis and treatment. Here, we present a 44-year-old female patient who presented to our hospital with complaints of abdominal pain, nausea, vomiting, anorexia and weight loss. After diagnostic investigations with laboratory and imaging methods, she was initially hospitalized with a preliminary diagnosis of cholangiocellular carcinoma (CCC). However, after a full work-up, the patient was diagnosed with Fasciola hepatica via ERCP and parasites were extracted with ERCP at the same time and then treated with a single dose of triclabendazole 10 mg/kg. Two months later, the clinical status of the patient had improved markedly, with resolution of all symptoms and all laboratory and imaging tests returning to within normal limits.

Human case of fasciolosis in Serbia treated with triclabendazole

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Pavlović Milorad

2014-01-01

Full Text Available Introduction. The number of humans infected by Fasciola hepatica is increasing worldwide. Humans can become accidental hosts by ingesting drinking water or plants contaminated with metacercariae. Case report. We reported a case of a 68-year-old Serbian woman, in which the diagnosis of acute fasciolosis had been established after serious diagnostic concerns. Based on clinical picture (episodic right upper quadrant abdominal pain, febrility and generalized body pain and biochemical analyses (high eosinophilia and high activity of alkaline phosphatase, she was appointed as suspected to the acute fasciolosis. Stool and duodenal aspirate exams were negative for Fasciola ova. In the absence of adequate serologic diagnostic for fasciolosis in Serbia, the diagnosis was confirmed using enzyme immunoassays and immunoblot at the Institute for Tropical Diseases in Hamburg, Germany. Soon after triclabendazole was administered, the symptoms disappeared and biochemical values returned to normal. Conclusion. The diagnosis of human fasciolosis may be problematic and delayed, especially in non endemic areas, because physicians rarely encounter this disease and a long list of other diseases must be considered in the differential diagnosis. The syndrome of eosinophilia, fever, and right upper quadrant abdominal pain suggest acute fasciolosis. Unclear source does not rule out fasciolosis.

Metazoan parasites of Plagioscion squamosissimus, an invasive species in the Tietê River, São Paulo, Brazil

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Ivan Moura Lapera

Full Text Available Abstract This study focused on the characterization and analysis of communities and infra-communities of metazoan parasites of Plagioscion squamosissimus caught in Promissão Reservoir in the Tietê River in Borborema (21°39′58”S, 49°8′49”W, state of São Paulo, Brazil. Fifty adult specimens caught by professional fishermen in March 2015 were necropsied. The fish presented an average standard length of 25.2 ± 2.2 cm and average weight of 328.82 ± 89.03 g. A total of 5,227 specimens of metazoan parasites were collected: 2,880 (55.1% adult Diplectanum piscinarius (Monogenoidea: Diplectanidae and 2,347 (44.9% Austrodiplostomum compactum metacercariae (Digenea, Diplostomidae, both with 100% prevalence and mean abundance of 57.6 and 46.9, respectively. Parasite diversity was low (species richness = 2, with a Simpson index (D equal to 0.505, and low values of Brillouin (HB = 0.687 and Margalef diversity (I = 0.117 indices. Berger-Parker’s index of dominance (p = 0.551 indicated a slight dominance of the monogenean parasite D. piscinarius. There was a positive correlation, assessed by Pearson coefficient between parasite abundance of D. piscinarius and standard length (r = 0.43 and weight (r = 0.51 of hosts.

Differential tolerances to ocean acidification by parasites that share the same host.

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MacLeod, C D; Poulin, R

2015-06-01

Ocean acidification is predicted to cause major changes in marine ecosystem structure and function over the next century, as species-specific tolerances to acidified seawater may alter previously stable relationships between coexisting organisms. Such differential tolerances could affect marine host-parasite associations, as either host or parasite may prove more susceptible to the stressors associated with ocean acidification. Despite their important role in many ecological processes, parasites have not been studied in the context of ocean acidification. We tested the effects of low pH seawater on the cercariae and, where possible, the metacercariae of four species of marine trematode parasite. Acidified seawater (pH 7.6 and 7.4, 12.5 °C) caused a 40-60% reduction in cercarial longevity and a 0-78% reduction in metacercarial survival. However, the reduction in longevity and survival varied distinctly between parasite taxa, indicating that the effects of reduced pH may be species-specific. These results suggest that ocean acidification has the potential to reduce the transmission success of many trematode species, decrease parasite abundance and alter the fundamental regulatory role of multi-host parasites in marine ecosystems. Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Occurrence of black-spot disease caused by metacercariae of Posthodiplostomum cuticola among juvenile fishes in water bodies in the Morava River basin

Czech Academy of Sciences Publication Activity Database

Ondračková, Markéta; Bartošová, Šárka; Valová, Zdenka; Jurajda, Pavel; Gelnar, M.

2004-01-01

Roč. 49, č. 3 (2004), s. 222-227 ISSN 1230-2821 R&D Projects: GA ČR GA524/02/0924 Institutional research plan: CEZ:AV0Z6093917 Keywords : Digenea * juvenile fish * host susceptibility Subject RIV: EG - Zoology Impact factor: 0.560, year: 2004 http://www.actaparasitologica.pan.pl/archive/49/toc49130.html

Vaccine potential of recombinant cathepsin B against Fasciola gigantica.

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Chantree, Pathanin; Phatsara, Manussabhorn; Meemon, Krai; Chaichanasak, Pannigan; Changklungmoa, Narin; Kueakhai, Pornanan; Lorsuwannarat, Natcha; Sangpairoj, Kant; Songkoomkrong, Sineenart; Wanichanon, Chaitip; Itagaki, Tadashi; Sobhon, Prasert

2013-09-01

In Fasciola gigantica, cathepsin Bs, especially cathepsin B2 and B3 are expressed in early juvenile stages, and are proposed to mediate the invasion of host tissues. Thus they are thought to be the target vaccine candidates that can block the invasion and migration of the juvenile parasite. To evaluate their vaccine potential, the recombinant cathepsin B2 (rFgCatB2) and cathepsin B3 (rFgCatB3) were expressed in yeast, Pichia pastoris, and used to immunize mice in combination with Freund's adjuvant to evaluate the protection against the infection by F. gigantica metacercariae, and the induction of immune responses. Mice immunized with both recombinant proteins exhibited high percent of parasite reduction at 60% for rFgCatB2 and 66% for rFgCatB3. Immunization by both antigens induced continuously increasing levels of IgG1 and IgG2a with a higher level of IgG1 isotype, indicating the mixed Th1/Th2 responses with Th2 predominating. When examined individually, the higher levels of IgG1 and IgG2a were correlated with the lower numbers of worm recoveries. Thus, both cathepsin B2 and cathepsin B3 are plausible vaccine candidates whose potential should be further tested in large economic animals. Copyright © 2013 Elsevier Inc. All rights reserved.

Prevalence of Fasciola species in ruminants under extensive management system in Ibadan southwestern Nigeria.

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Adediran, O A; Adebiyi, A I; Uwalaka, E C

2014-12-01

Fasciolosis is a zoonotic trematode of ruminants and a common parasite in cattle in Nigeria. With the increasing numbers of small household cattle herds kept under extensive management system as well as the more usual small ruminants, there is a possibility therefore that parasites common to these ruminants but better adapted to cattle such as Fasciola may also be on the increase within the small ruminant population as they graze freely over the same wide area and sometimes freely mix. The aim of the study is to determine the prevalence of Fasciola spp in sheep, goats and cattle in Ibadan during the short wet season, (August-October 2011). Faecal samples were collected per rectum from 440 each of goats and sheep and 291 cattle. Helminth ova were recovered using sedimentation technique for faecal examination. A total of 40 goats (9.1%), 53 sheep (12.0%) and 158 cattle (54.3%) were positive for Fasciola ova which reflects an increase in prevalence in small ruminants. This implies that the entire Ibadan is endemic for Fasciola spp and small ruminants may serve as an important link in the epidemiology of Fasciola spp infection. More so, this has implications for the scavenging household animals which receive little or no medical attention and man that may acquire this trematode by inadvertently eating vegetables with encysted Fasciola metacercariae.

THE BIOTIC FACTOR OF TREMATOD OPISTHORHIS FELINEUS INVASION INFLUENCE ON HOST IMMUNE STATUS AND SOMATIC CELLS PROLIFERATIVE ACTIVITY

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A. G. Rybka

2016-01-01

Full Text Available The paper confirms long-time opisthorhis invasion role as a risk factor of host immune system reconstitution as well as an important factor in holangiocarcinomas development. It was shown that opisthorhosis invasion primal stage induce host immune system reconstitution. Host immune B-cells system is activated by metacercaria antigens, while the same antigens inhibits T-cells activity. Opisthorhis metabolites stimulate proliferative mithogen-induced T-cells acti vity. Chronic opisthorchis invasion leads to immune system disbalance. It means: decrease of specific and non-speci fic natural killers activity, number of high proliferative activity T-lymphocytes and the shift of regulatory T-cells subset to suppressors prevalence. At the same time specific as well as non-specific T-suppressors functional ability is very low. It was shown T-cells helper-amplifier activation. Despite of circulating B-cells decrease the antibody produced cells number is spleen increases significantly at the same time with circulating immune complexes accumulation. Even 3–6 month after dehelmintisation the immune system disbalance decreases but lefts. In addition, chronic opisthorhis invasion leads to the proliferative processes activation in ductal epithelium, liver, lymph nodes and in other organs which leads to cancer proliferation. According to the results obtained the opisthorhis infected patients needs to be immunocorrected before as well as after dehelmintisation for holangiocancerogenesis profylaxis.

Host differences in response to trickle infection with Fasciola gigantica in buffalo, Ongole and Bali calves.

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Wiedosari, E; Hayakawa, H; Copeman, B

2006-01-01

Progressive weight gain, faecal egg counts, packed cell volume, percent eosinophils in blood, serum antibody and serum levels of glutamate dehydrogenase and gamma-glutamyl transpeptidase were recorded in seven swamp buffalo (Bubalis bubalis), 7 Ongole (Bos indicus) and four Bali calves (Bos sundiacus) which were infected orally with 15 metacercariae of Fasciola gigantica twice weekly for 32 weeks. Similar observations were made on four buffalo, 4 Ongole calves and 3 Bali calves maintained fluke-free as controls. Flukes were counted at slaughter 36 weeks after initial infection. Mean daily weight gains of infected Bali (228 +/- 100 (SD) g/day) and infected Ongole calves (328 +/- 57 (SD) g/day) were lower (p = 0.026 and 0.067, respectively) than those of control calves (405 +/- 107 (SD) g/day), but infected buffalo calves (379 +/- 78 (SD) g/day) had similar weight gains to those of the controls (p = 0.57). Throughout the trial, faecal Fasciola egg counts in buffaloes were about one-fifth of counts of Ongole calves, and counts in Bali calves were intermediate. Ongole calves had three times the number of flukes at slaughter in their liver compared to buffalo and Bali calves, which had similar numbers. However, there was evidence that Bali calves had acquired a degree of resistance about 24 weeks after infection commenced and may have lost adult flukes as a consequence.

An ecological and comparative analysis of parasites in juvenile Mugil liza (Pisces, Mugilidae from two sites in Samborombón bay, Argentina

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Martin M. Montes

2015-12-01

Full Text Available ABSTRACT Mugil liza Valenciennes, 1836 is an economically important food fish and has been recommended for aquaculture in South America. A total of 278 fishes were collected in the spring and summer of 2009 and 2010. These fish were sorted into sample groups according to their size class. We used Bayesian statistics and 95% credible intervals for each parameter tested were calculated. Fish studied harbored a total of 15 different species of parasites. Diversity of parasite species found on Mugil liza was greatest at the S.R.C. collection site, but evidenced a lower species richness than at A.R. site. The 1st size fishes of both sites evidenced greater parasite diversity than either 2nd or 3rd size fish. Differences observed could be explained by the different use of habitat types at the two sites or differential susceptibility to infection by parasites. The dominance of D. fastigatainfluenced observed results of lower community diversity indexes. New works elucidating different parasite life cycles within juvenile and adults ofM. liza in Argentina, promise to be important for determining the risk of the parasitism by zoonotic metacercariae A. (P. longa and use of this fish as food and an economic resource, and the possible use of mullet parasites in other promising fields as indicators of biodiversity, and/ or water contamination.

Current status of Paragonimus and paragonimiasis in Ecuador

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Manuel Calvopiña

2014-11-01

Full Text Available A review of national and international publications on paragonimiasis in Ecuador, epidemiological records from the Ministry of Public Health and unpublished research data was conducted to summarise the current status of the parasite/disease. The purpose of the review is to educate physicians, policy-makers and health providers on the status of the disease and to stimulate scientific investigators to conduct further research. Paragonimiasis was first diagnosed in Ecuador 94 years ago and it is endemic to both tropical and subtropical regions in 19 of 24 provinces in the Pacific Coast and Amazon regions. Paragonimus mexicanus is the only known species in the country, with the mollusc Aroapyrgus colombiensis and the crabs Moreirocarcinus emarginatus, Hypolobocera chilensis and Hypolobocera aequatorialis being the primary and secondary intermediate hosts, respectively. Recent studies found P. mexicanus metacercariae in Trichodactylus faxoni crabs of the northern Amazon. Chronic pulmonary paragonimiasis is commonly misdiagnosed and treated as tuberculosis and although studies have demonstrated the efficacy of praziquantel and triclabendazole for the treatment of human infections, neither drug is available in Ecuador. Official data recorded from 1978-2007 indicate an annual incidence of 85.5 cases throughout the 19 provinces, with an estimated 17.2% of the population at risk of infection. There are no current data on the incidence/prevalence of infection, nor is there a national control programme.

Review and Current Status of Opisthorchis viverrini Infection at the Community Level in Thailand.

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Kaewpitoon, Natthawut; Kootanavanichpong, Nusorn; Kompor, Ponthip; Chavenkun, Wasugree; Kujapun, Jirawoot; Norkaew, Jun; Ponphimai, Sukanya; Matrakool, Likit; Tongtawee, Taweesak; Panpimanmas, Sukij; Rujirakul, Ratana; Padchasuwan, Natnapa; Pholsripradit, Poowadol; Eksanti, Thawatchai; Phatisena, Tanida; Loyd, Ryan A; Kaewpitoon, Soraya J

2015-01-01

Opisthorchis viverrini is remains a public health problem in Thailand, particularly in the northeast and north regions which have the highest incidences of chonalgiocarcinoma (CCA). O. viverrini causes the disease opithorchiasis, and its has been classified as a group 1 biological carcinogen. Humans, dogs, and cats become infected with O. viverrini by ingesting raw or undercooked fish containing infective metacercariae. The first human cases of O. viverrini infection were reported in Thailand 100 years ago, and it's still a problem at the community level. Based on data for the year 2009, more than 6 million people were infected with O. viverrini. Associated medical care and loss of wages in Thailand costs about $120 million annually. This review highlights the current status of O. viverrini infection in communities of Thailand through active surveillance for the five years period from 2010 and 2015. A total of 17 community-based surveys were conducted, most in the northeast region. Some 7 surveys demonstrated a high prevalence over 20%, and the highest was 45.7%. Most commonly infection was found in age group of 35 years and older, males, and agricultural workers. Although, the national prevalence may be decreasing but the results show that the O. viverrini infection is still high in communities of the northeast region. Therefore, the focus in populations living in northeast Thailand should be screening of infection and changing their eating behavior.

The numbers game: quantitative analysis of Neorickettsia sp. propagation through complex life cycle of its digenean host using real-time qPCR.

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Greiman, Stephen E; Tkach, Vasyl V

2016-07-01

Bacteria of the genus Neorickettsia are obligate intracellular endosymbionts of parasitic flukes (Digenea) and are passed through the entire complex life cycle of the parasite by vertical transmission. Several species of Neorickettsia are known to cause diseases in domestic animals, wildlife, and humans. Quantitative data on the transmission of the bacteria through the digenean life cycle is almost completely lacking. This study quantified for the first time the abundance of Neorickettsia within multiple stages of the life cycle of the digenean Plagiorchis elegans. Snails Lymnaea stagnalis collected from a pond in North Dakota were screened for the presence of digenean cercariae, which were subsequently tested for the presence of Neorickettsia. Three L. stagnalis were found shedding P. elegans cercariae infected with Neorickettsia. These snails were used to initiate three separate laboratory life cycles and obtain all life cycle stages for bacterial quantification. A quantitative real-time PCR assay targeting the GroEL gene was developed to enumerate Neorickettsia sp. within different stages of the digenean life cycle. The number of bacteria significantly increased throughout all stages, from eggs to adults. The two largest increases in number of bacteria occurred during the period from eggs to cercariae and from 6-day metacercariae to 48-h juvenile worms. These two periods seem to be the most important for Neorickettsia propagation through the complex digenean life cycle and maturation in the definitive host.

Ecology of parasites of Metynnis lippincottianus (Characiformes: Serrasalmidae from the eastern Amazon region, Macapá, State of Amapá, Brazil

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Maria Danielle Figueiredo Guimarães Hoshino

2014-05-01

Full Text Available The present study provides the first investigation on the ecological aspects of the parasites M. lippincottianusfrom the Amazonian basin, as well as the parasite-host relationship. 76 out of the examined fish (98.7% were parasitized by at least one species of parasites. A total of 8,774 parasites were collected, being Ichthyophthirius multifiliis, Anacanthorus jegui, Dadayius pacupeva, Digenea gen. sp. (metacercariae, Procamallanus (Spirocamallanus inopinatus, Procamallanus (Spirocamallanus sp., Spinoxyuris oxydoras, Contracaecum sp. larvae, Dolops longicauda and Hirudinea gen. sp. Ichthyophthirius multifiliis was the dominant species, followed by A. jegui. Among the endoparasites, S. oxydoras and D. pacupeva were predominant. The mean diversity of parasites was HB = 0.96 ± 0.32 and there was aggregate distribution pattern. A positive correlation of body weight with the abundance of I. multifiliis and S. Oxydoras was observed, whereas a negative correlation of body weight with abundance of the Contracaecum sp. larvae was found. The relative condition factor (Kn was not negatively affected by parasites, and a positive correlation between Kn and abundance of I. multifiliis, S. oxydoras and D. pacupeva was found. This study is the first one to record I. multifiliis, D. longicauda and A. jegui parasitizing M. lippincottianus, as well as the first record of D. pacupeva and S. oxydoras in the Amazonas river system.

Immune responses induced by co-infection with Capillaria hepatica in Clonorchis sinensis-infected rats.

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Moon, E-K; Lee, S-H; Goo, T W; Quan, F-S

2018-07-01

Clonorchis sinensis and Capillaria hepatica are zoonotic parasites that mainly infect the liver and cause serious liver disorders. However, immunological parameters induced by co-infection with these parasites remain unknown. In this study, for the first time, we investigated immunological profiles induced by co-infection with C. hepatica (CH) in C. sinensis (CS)-infected rats (Sprague-Dawley). Rats were infected primarily with 50 metacercariae of C. sinensis; 4 weeks later, they were subsequently infected with 1000 infective C. hepatica eggs. Significantly higher levels of C. sinensis- or C. hepatica-specific IgG antibodies were found in the sera of rats. Interestingly, no cross-reacting antibody was observed between C. sinensis and C. hepatica infections. Significantly raised eosinophil levels were found in the blood of C. sinensis/C. hepatica co-infected rats (CS + CH) compared to the blood of rats infected singly with C. sinensis. Co-infected rats showed significantly higher levels of lymphocyte proliferation and cytokine production compared to a single C. sinensis infection. The worm burden of C. sinensis was significantly reduced in co-infected rats compared to the single C. sinensis infection. These results indicate that the eosinophils, lymphocyte proliferation and cytokine production induced by subsequent infection with C. hepatica in C. sinensis-infected rats might contribute to the observed C. sinensis worm reduction.

Variations in local adaptation of allopatric Fasciola hepatica to French Galba truncatula in relation to parasite origin.

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Dar, Y; Lounnas, M; Djuikwo Teukeng, F F; Mouzet, R; Courtioux, B; Hurtrez-Boussès, S; Vignoles, P; Dreyfuss, G; Rondelaud, D

2013-07-01

Two French populations of Galba truncatula were subjected to experimental infections with Egyptian and French isolates of Fasciola sp. miracidia, originating from cattle and sheep, to compare characteristics of snail infections in allopatric and sympatric groups. All sampled Egyptian isolates were identified as Fasciola hepatica using microsatellite markers. Compared to snails infected with French miracidia, snail survival at day 30 post-exposure was significantly greater in the Egyptian groups, while prevalence of infection was significantly lower (in an Egyptian group infected with cattle-derived miracidia) or did not show any significant differences in the other three cases. The total number of metacercariae was significantly higher in the four Egyptian groups. However, snail population and the mammalian origin of F. hepatica had also a significant effect on this parameter. The dissection of snail cadavers showed a significantly higher number of free rediae in the Egyptian groups, even if snail population also had a significant effect on the redial burden. Both Egyptian isolates of F. hepatica could easily develop in French snails, causing a low mortality in snails and inducing a metacercarial production higher than that noted in sympatric infections. However, the mammalian origin of F. hepatica eggs and the quality of snail populations as intermediate hosts had to be taken into account for studying local adaptation in reason of their effects on this process.

Studies on the epidemiology and histopathology of Euclinostomum heterostomum (Trematoda; Digenea infection in Channa punctata from North India

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Shareef P. A. Ahammed

2015-09-01

Full Text Available A survey on the occurrence and epidemiology of the encysted progenetic metacercariae of Euclinostomum heterostomum infection in Channa punctata in the Aligarh region of North India revealed a mean prevalence, intensity, and abundance of 18.61, 1.52, and 0.38%, respectively, during the period from April 2011 to March 2012. Liver, kidney, peritoneum, muscle, and ovary were found to be infected with this parasite, and the later three are reported for the first time in this fish species. The histopathology of the infected tissues indicated the following at the host-parasite interface: tissue damage, infiltration of immune cells into the cyst wall, chronic inflammatory responses, and granulomatous lesions. The infected liver showed degeneration of hepatocytes, cytoplasmic vacuolation, nuclear alterations, mallory body formation, fibrosis, and necrosis. The pathology of the infected kidney included distortion and dilation of renal tubules, vacuolar degeneration, hypertrophy and hyperplasia of tubular epithelial cells, occlusion of tubules, fibrosis, hemorrhage, and congestion of glomeruli. The infected muscle demonstrated comparatively fewer pathological changes confined only to the circumference of the cyst wall. The ovary displayed the least changes. The conclusions drawn from the study are that the large metacercarial cysts formed by E. heterostomum in the vital organs of the economically important fish C. punctata could result in the impairment of fish physiology and health, thereby affecting their productivity and quality for human consumption.

Isolation and characterization of Cu/Zn-superoxide dismutase in Fasciola gigantica.

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Lalrinkima, H; Raina, O K; Chandra, Dinesh; Jacob, Siju Susan; Bauri, R K; Chandra, Subhash; Yadav, H S; Singh, M N; Rialch, A; Varghese, A; Banerjee, P S; Kaur, Navneet; Sharma, Arvind

2015-01-01

A full-length complementary DNA (cDNA) encoding Cu/Zn-superoxide dismutase was isolated from Fasciola gigantica that on nucleotide sequencing showed a close homology (98.9%) with Cu/Zn-superoxide dismutase (SOD) of the temperate liver fluke, F. hepatica. Expression of the gene was found in all the three developmental stages of the parasite viz. adult, newly excysted juvenile and metacercaria at transcriptional level by reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. F. gigantica Cu/Zn-SOD cDNA was cloned and expressed in Escherichia coli. Enzyme activity of the recombinant protein was determined by nitroblue tetrazolium (NBT)-polyacrylamide gel electrophoresis (PAGE) and this activity was inactivated by hydrogen peroxide but not by sodium azide, indicating that the recombinant protein is Cu/Zn-SOD. The enzyme activity was relatively stable at a broad pH range of pH 4.0-10.0. Native Cu/Zn-superoxide dismutase protein was detected in the somatic extract and excretory-secretory products of the adult F. gigantica by Western blotting. NBT-PAGE showed a single Cu/Zn-SOD present in the somatic extract while three SODs are released ex vivo by the adult parasite. The recombinant superoxide dismutase did not react with the serum from buffaloes infected with F. gigantica. The role of this enzyme in defense by the parasite against the host reactive oxygen species is discussed. Copyright © 2015 Elsevier Inc. All rights reserved.

Variability in prevalence of Cymatocarpus solearis (Trematoda, Brachycoeliidae) in Caribbean spiny lobsters Panulirus argus (Decapoda: Palinuridae) from Bahía de la Ascensión (Mexico).

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Briones-Fourzán, Patricia; Muñoz de Cote-Hernández, Rubén; Lozano-Álvarez, Enrique

2016-06-01

Cymatocarpus solearis, a brachycoeliid trematode that parasitizes sea turtles, uses spiny lobsters Panulirus argus as second intermediate hosts. We examined variability in infection by C. solearis in Bahía de la Ascensión, Mexico, a tropical bay with a highly productive fishery for P. argus that is based on the extensive use of casitas (large artificial shelters), which can harbor multiple juveniles and adults of this gregarious lobster species. We sampled 3956 lobsters from 466 casitas distributed over three bay zones within two closed seasons and one fishing season. In these lobsters (9.5-115.2mm carapace length, CL), the average infection prevalence was 11.7% but the probability of infection increased significantly with size. Prevalence varied with season but was consistently higher in one zone than in the other two zones. Infection with C. solearis was negatively related with clinical infection with Panulirus argus Virus 1 (PaV1). We also sampled 405 lobsters from the commercial catch obtained throughout the bay at the onset of the fishing season. In these relatively larger lobsters (70.3-168.0mm CL), average prevalence of C. solearis was higher (23.5%) but was not affected by lobster size or sex. Encysted metacercariae occurred in both abdominal and cephalothoracic muscles. The effects of C. solearis on phenotypic traits of P. argus that may potentially impact the host population dynamics and fisheries remain to be investigated. Copyright © 2016 Elsevier Inc. All rights reserved.

[Epidemiological investigation on fascioliasis and its risk factors in population of Binchuan County, Yunnan Province].

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Chen, Feng; Liu, Yu-hua; Yang, Hui; Duan, Yii-chun; Yang, Jing; Zhao, Yin-jiao; Hao, Ming-ming; Pan, Li-juan; Deng, Yin-ju; Shi, Ya-nan; Chen, Fu-ling; Zhang, Jian-guo; Luo, Jia-jun

2015-08-01

To understand the infection status of Fasciola and related risk factors in population of Binchuan County, Yunnan Province. In February and March, 2014, 6 natural villages from 2 administrative villages in Zhoucheng Town were selected as investigated sites, and the permanent residents aged ≥ 5 years were examined by ELISA for the serum antibody detection, and then the positive ones were examined by Kato-Katz technique and the hatching method for Fasciola egg detection. Meanwhile, the fascioliasis cases that checked out by the above methods were further surveyed, and the residents who with the same gender, similar age and negative results in the blood examination were selected as the controls, then the risk factors were explored through a case control study. Totally 1 207 people were included in the blood examination, and 36 positives were found with a positive rate of 2.98%. Though the differences between the positive rates of the serum antibody in people with different genders and age were not statistically significant (χ2 = 1.686, 7.106, both P > 0.05) , those between the rates of people in different administrative villages and natural villages were statistically significant (χ2 = 4.103, 13.181, both P 0.05). There is still sporadic Fasciola infection in Binchuan County. Eating raw vegetables and drinking unboiled water which are contaminated by Fasciola metacercariae may be the infection routes of local people, and to find out the exact entrance way is the key points for the infection control.

Potential interactions between metazoan parasites of the Mayan catfish Ariopsis assimilis and chemical pollution in Chetumal Bay, Mexico.

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Vidal-Martínez, V M; Aguirre-Macedo, M L; Noreña-Barroso, E; Gold-Bouchot, G; Caballero-Pinzón, P I

2003-06-01

The effect of pollutants on the intensity of infection of metazoan parasites in the Mayan catfish, Ariopsis assimilis was investigated. Data were collected on pollutants and metazoan parasites from 76 catfish from five localities in Chetumal Bay in October, 1996. Nineteen pollutants (pesticides, polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs)) were found in the catfish livers. Heavy metal content was not determined. Nineteen metazoan parasite species were recovered. After controlling for fish length and sampling station, there was a significant negative linear relationship between the intensity of the larval digenean Mesostephanus appendiculatoides and 1,1,1,-trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) concentrations. This negative relationship may be explained either by the effect of the pesticide on the mortality of (i) free-living larval forms, (ii) metacercariae in the fish, (iii) infected fish or (iv) intermediate host snails. There were significant differences between fish parasitized and not parasitized with M. appendiculatoides with respect to their DDT concentrations. There were also significant differences between the variances of the mean Clark's coefficient of condition values between catfish parasitized and not parasitized by M. appendiculatoides, with the variance of non-parasitized catfish being significantly larger. The results provided statistical evidence that DDT has a detrimental effect on M. appendiculatoides infection intensity. Furthermore, the significantly larger variance value of Clark's coefficient for non-parasitized fish suggested that DDT affects both the parasite and general host condition.

Centrocestus formosanus (Opisthorchiida: Heterophyidae como causa de muerte de alevines de tilapia gris Oreochromis niloticus (Perciforme: Cichlidae en el Pacífico seco de Costa Rica

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Donald Arguedas Cortés

2010-12-01

Full Text Available Centrocestus formosanus es un parásito trematodo zoonótico originario de Asia asociado con muertes de peces principalmente de cultivo. 907 moluscos provenientes de estanques sembrados con tilapias, seleccionados uno por provincia fueron identificados al nivel taxonómico especifico. Se identificaron cuatro gastrópodos y un bivalvo: M. tuberculata, M. turricula, P. flagellata, H. cubensis y A. luteola. Se reporta, por primera vez, la presencia de dos especies de moluscos en Costa Rica. Se identificaron siete morfotipos de cercarias parasitando las cinco especies de moluscos encontradas. En la segunda exposición experimental se demostró que el morfotipo parapleurolofocercus encontrado en M. tuberculata concuerda con el hallazgo de C. formosanus en alevines de tilapia, después del examen clínico, anatomopatológico y parasitológico realizado a los alevines expuestos. Las metacercarias fueron extraídas del quiste utilizando microagujas y micropinzas lavadas en solución salina fisiológica (0.65%, fijadas en formol caliente al 4% y después esquematizadas con una cámara clara adaptada a un microscopio fotónico, estimándose una abundancia e intensidad media de 1018-1027 digeneos por branquia en cada pez parasitado, determinándose así el hospedador intermediario primario y secundario del parásito. En el presente trabajo se reporta por primera vez Centrocestus formosanus en Costa Rica.

Coproantígenos de Fasciola hepatica de posible utilidad en el diagnóstico de la fascioliasis Identification and isolation of coproantigens of Fasciola hepatica that are potentially useful in the diagnosis of fascioliasis

Directory of Open Access Journals (Sweden)

Ana M. Espino

2000-04-01

Full Text Available En el presente estudio se describen cuáles son los antígenos de Fasciola hepatica que están presentes en las heces de pacientes con fascioliasis crónica y de ratas infectadas experimentalmente con metacercarias de F. hepatica. Mediante el empleo de la técnica de inmunoelectrotransferencia (Western blot con un suero hiperinmune obtenido frente a antígenos de excreción/secreción de adultos de F. hepatica, se pudo demostrar que en las heces de los pacientes se encuentran antígenos de posible interés para el diagnóstico con masas moleculares de 14, 19, 20, 23, 25, 32, 46, 51 y 62 kilodaltons (kDa. Además, se comprobó que algunos de estos péptidos (los de 14, 20, 23 y 51 kDa son reconocidos también por la mayoría de los sueros de pacientes crónicos. Después de purificar por cromatografía de afinidad los antígenos presentes en heces de ratas con 6 a 12 semanas de infección, empleando para ello el anticuerpo monoclonal ES78 acoplado a Sepharosa 4B-CNBr, se identificaron seis polipéptidos de 11, 14, 26, 32, 47 y 51 kDa; otros tres polipéptidos, de 17, 24 y 66 kDa, solo se pudieron identificar en heces de ratas con 10 a 12 semanas de infección. Nuestros resultados sugieren que estos polipéptidos pueden ser antígenos comunes a ambos estadios parasitarios y constituir importantes marcadores de la fascioliasis aguda y crónica, en particular los de 14, 24, 26 y 51 kDa, puesto que reaccionaron con el inmunosuero, los sueros humanos y el anticuerpo monoclonal ES78.This study describes which antigens of Fasciola hepatica are present in the feces of patients with chronic fascioliasis and in the feces of rats infected experimentally with F. hepatica metacercariae. Using a Western blot assay technique with hyperimmune serum obtained from excretory-secretory antigens of adult F. hepatica, we found in the patients' feces antigens of possible diagnostic interest, with molecular weights of 14, 19, 20, 23, 25, 32, 46, 51, and 62 kilodaltons

Protective role of purified cysteine proteinases against Fasciola gigantica infection in experimental animals.

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El-Ahwany, Eman; Rabia, Ibrahim; Nagy, Faten; Zoheiry, Mona; Diab, Tarek; Zada, Suher

2012-03-01

Fascioliasis is one of the public health problems in the world. Cysteine proteinases (CP) released by Fasciola gigantica play a key role in parasite feeding, migration through host tissues, and in immune evasion. There has been some evidence from several parasite systems that proteinases might have potential as protective antigens against parasitic infections. Cysteine proteinases were purified and tested in vaccine trials of sheep infected with the liver fluke. Multiple doses (2 mg of CP in Freund's adjuvant followed by 3 booster doses 1 mg each at 4 week intervals) were injected intramuscularly into sheep 1 week prior to infect orally with 300 F. gigantica metacercariae. All the sheep were humanely slaughtered 12 weeks after the first immunization. Changes in the worm burden, ova count, and humoral and cellular responses were evaluated. Significant reduction was observed in the worm burden (56.9%), bile egg count (70.7%), and fecel egg count (75.2%). Immunization with CP was also found to be associated with increases of total IgG, IgG(1), and IgG(2) (P<0.05). Data showed that the serum cytokine levels of pro-inflammatory cytokines, IL-12, IFN-γ, and TNF-α, revealed significant decreases (P<0.05). However, the anti-inflammatory cytokine levels, IL-10, TGF-β, and IL-6, showed significant increases (P<0.05). In conclusion, it has been found that CP released by F. gigantica are highly important candidates for a vaccine antigen because of their role in the fluke biology and host-parasite relationships.

Serum levels of cytokines in water buffaloes experimentally infected with Fasciola gigantica.

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Zhang, Fu-Kai; Guo, Ai-Jiang; Hou, Jun-Ling; Sun, Miao-Miao; Sheng, Zhao-An; Zhang, Xiao-Xuan; Huang, Wei-Yi; Elsheikha, Hany M; Zhu, Xing-Quan

2017-09-15

Fasciola gigantica infection in water buffaloes causes significant economic losses especially in developing countries. Although modulation of the host immune response by cytokine neutralization or vaccination is a promising approach to control infection with this parasite, our understanding of cytokine's dynamic during F. gigantica infection is limited. To address this, we quantified the levels of serum cytokines produced in water buffaloes following experimental infection with F. gigantica. Five buffaloes were infected via oral gavage with 500 viable F. gigantica metacercariae and blood samples were collected from buffaloes one week before infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples were simultaneously determined using ELISA. F. gigantica failed to elicit the production of various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. The results also revealed suppression of the immune responses as a feature of chronic F. gigantica infection in buffaloes. Taken together, F. gigantica seems to elicit a modest Th2 response at early stage of infection in order to downregulate harmful Th1- and Th17-type inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. gigantica immune response and its relation to pathogenesis requires further study. Copyright © 2017 Elsevier B.V. All rights reserved.

Variation of parasite load and immune parameters in two species of New Zealand shore crabs.

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Dittmer, Jessica; Koehler, Anson V; Richard, Freddie-Jeanne; Poulin, Robert; Sicard, Mathieu

2011-09-01

While parasites are likely to encounter several potential intermediate hosts in natural communities, a parasite's actual range of compatible hosts is limited by numerous biological factors ranging from behaviour to immunology. In crustaceans, two major components of immunity are haemocytes and the prophenoloxidase system involved in the melanisation of foreign particles. Here, we analysed metazoan parasite prevalence and loads in the two sympatric crab species Hemigrapsus crenulatus and Macrophthalmus hirtipes at two sites. In parallel, we analysed the variation in haemocyte concentration and amount of circulating phenoloxidase (PO) in the haemolymph of the same individuals in an attempt to (a) explain differences in parasite prevalence and loads in the two species at two sites and (b) assess the impact of parasites on these immune parameters. M. hirtipes harboured more parasites but also exhibited higher haemocyte concentrations than H. crenulatus independent of the study site. Thus, higher investment in haemocyte production for M. hirtipes does not seem to result in higher resistance to parasites. Analyses of variation in immune parameters for the two crab species between the two sites that differed in parasite prevalence showed common trends. (a) In general, haemocyte concentrations were higher at the site experiencing higher parasitic pressure while circulating PO activity was lower and (b) haemocyte concentrations were influenced by microphallid trematode metacercariae in individuals from the site with higher parasitic pressure. We suggest that the higher haemocyte concentrations observed in both crab species exposed to higher parasitic pressure may represent an adaptive response to the impact of parasites on this immune parameter.

Alaria mesocercariae in the tails of red-sided garter snakes: evidence for parasite-mediated caudectomy.

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Uhrig, Emily J; Spagnoli, Sean T; Tkach, Vasyl V; Kent, Michael L; Mason, Robert T

2015-12-01

Trematodes of the genus Alaria develop into an arrested stage, known as mesocercariae, within their amphibian second intermediate host. The mesocercariae are frequently transmitted to a non-obligate paratenic host before reaching a definitive host where further development and reproduction can occur. Snakes are common paratenic hosts for Alaria spp. with the mesocercariae often aggregating in the host's tail. In the current study, we used morphological examination and molecular analyses based on partial sequences of nuclear large ribosomal subunit gene and mitochondrial cytochrome C oxidase subunit 1 gene to identify larvae in the tails of red-sided garter snakes (Thamnophis sirtalis parietalis) as mesocercariae of Alaria marcianae, Alaria mustelae, and Alaria sp. as well as metacercariae of Diplostomidae sp. of unknown generic affiliation. We assessed infection prevalence, absolute and relative intensity, and associated pathological changes in these snakes. Infection prevalence was 100 % for both male and female snakes. Infection intensity ranged from 11 to more than 2000 mesocercariae per snake tail but did not differ between the sexes. Gross pathological changes included tail swelling while histopathological changes included mild inflammation and the presence of mucus-filled pseudocysts surrounding mesocercariae, as well as the compression and degeneration of muscle fibers. Our results indicate that mesocercariae can lead to extensive muscle damage and loss in both sexes which likely increases the fragility of the tail making it more prone to breakage. As tail loss in garter snakes can affect both survival and reproduction, infection by Alaria mesocercariae clearly has serious fitness implications for these snakes.

Increased hepatic Th2 and Treg subsets are associated with biliary fibrosis in different strains of mice caused by Clonorchis sinensis

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Fang, Fan; Du, Ying; Ma, Rui; Li, Xiang-Yang; Yu, Qian; Meng, Di; Tang, Ren-Xian; Zheng, Kui-Yang

2017-01-01

Previous studies showed that CD4+T cells responses might be involved in the process of biliary fibrosis. However, the underlying mechanism resulting in biliary fibrosis caused by Clonorchis sinensis remains not yet fully elucidated. The objectives of the present study were to investigate the different profiles of hepatic CD4+T cell subsets (Th1, Th2, Th17 and Treg cells) and their possible roles in the biliary fibrosis of different strains of mice (C57BL/6, BALB/c and FVB mice) induced by C. sinensis infection. C57BL/6, BALB/c and FVB mice were orally gavaged with 45 metacercariae. All mice were sacrificed on 28 days post infection in deep anesthesia conditions. The leukocytes in the liver were separated to examine CD4+T cell subsets by flow cytometry and the left lobe of liver was used to observe pathological changes, collagen depositions and the concentrations of hydroxyproline. The most serious cystic and fibrotic changes appeared in FVB infected mice indicated by gross observation, Masson’s trichrome staining and hydroxyproline content detection. In contrast to C57BL/6 infected mice, diffuse nodules and more intensive fibrosis were observed in the BALB/c infected mice. No differences of the hepatic Th1 subset and Th17 subset were found among the three strains, but the hepatic Th2 and Treg cells and their relative cytokines were dramatically increased in the BALB/c and FVB infected groups compared with the C57BL/6 infected group (Psinensis-infected mice. PMID:28151995

Clonorchis sinensis and Opisthorchis spp. in Vietnam: current status and prospects.

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Doanh, Pham N; Nawa, Yukifumi

2016-01-01

Clonorchis sinensis and Opisthorchis viverrini are clinically important small liver flukes because of their known association with development of cholangiocarcinoma. In Vietnam, high prevalence of C. sinensis infection in humans was previously reported in northern provinces, and O. viverrini infection has been detected in several central provinces. However, diagnosis of C. sinensis and O. viverrini infections in the past was merely based on faecal egg examination. This method alone can lead to misidentification at the species level because of morphological similarity between the eggs of these liver flukes and minute intestinal trematodes of the family Heterophyidae. In fact, recent surveys in Vietnam revealed that infection with several minute intestinal flukes, such as Haplorchis pumilio and H. taichui, are much more common than infection with C. sinensis or O. viverrini, and they often co-infect humans. Thus, previously reported prevalence of small liver fluke infection in Vietnam was likely over-estimated due to mis identification of parasites in copro-parasitological examinations. In addition, there is some confusion about identification of cercariae, metacercariae and also adults of C. sinensis and O. viverrini in intermediate and definitive hosts. The aim of this review is, therefore, to draw realistic pictures of the past and present scientific reports on the epidemiology and biology of C. sinensis and Opisthorchis spp. infection in Vietnam. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Mortality of centrarchid fishes in the Potomac drainage: survey results and overview of potential contributing factors.

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Blazer, V S; Iwanowicz, L R; Starliper, C E; Iwanowicz, D D; Barbash, P; Hedrick, J D; Reeser, S J; Mullican, J E; Zaugg, S D; Burkhardt, M R; Kelble, J

2010-09-01

Skin lesions and spring mortality events of smallmouth bass Micropterus dolomieu and selected other species were first noted in the South Branch of the Potomac River in 2002. Since that year morbidity and mortality have also been observed in the Shenandoah and Monocacy rivers. Despite much research, no single pathogen, parasite, or chemical cause for the lesions and mortality has been identified. Numerous parasites, most commonly trematode metacercariae and myxozoans; the bacterial pathogens Aeromonas hydrophila, Aeromonas salmonicida, and Flavobacterium columnare; and largemouth bass virus have all been observed. None have been consistently isolated or observed at all sites, however, nor has any consistent microscopic pathology of the lesions been observed. A variety of histological changes associated with exposure to environmental contaminants or stressors, including intersex (testicular oocytes), high numbers of macrophage aggregates, oxidative damage, gill lesions, and epidermal papillomas, were observed. The findings indicate that selected sensitive species may be stressed by multiple factors and constantly close to the threshold between a sustainable (healthy) and nonsustainable (unhealthy) condition. Fish health is often used as an indicator of aquatic ecosystem health, and these findings raise concerns about environmental degradation within the Potomac River drainage. Unfortunately, while much information has been gained from the studies conducted to date, due to the multiple state jurisdictions involved, competing interests, and other issues, there has been no coordinated approach to identifying and mitigating the stressors. This synthesis emphasizes the need for multiyear, interdisciplinary, integrative research to identify the underlying stressors and possible management actions to enhance ecosystem health.

Analysis of the expression and antioxidant activity of 2-Cys peroxiredoxin protein in Fasciola gigantica.

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Sangpairoj, Kant; Changklungmoa, Narin; Vanichviriyakit, Rapeepun; Sobhon, Prasert; Chaithirayanon, Kulathida

2014-05-01

2-Cys peroxiredoxin (Prx) is the main antioxidant enzyme in Fasciola species for detoxifying hydrogen peroxide which is generated from the hosts' immune effector cells and the parasites' own metabolism. In this study, the recombinant Prx protein from Fasciola gigantica (rFgPrx-2) was expressed and purified in a prokaryotic expression system. This recombinant protein with molecular weight of 26 kDa was enzymatically active in reduction of hydrogen peroxide both in presence of thioredoxin and glutathione systems, and also protected the supercoiled plasmid DNA from oxidative damage in metal-catalyzed oxidation (MCO) system in a concentration-dependent manner. By immunoblotting, using antibody against rFgPrx-2 as probe, a native FgPrxs, whose MW at 25 kDa, was detected in all developmental stages of the parasite. Concentrations of native FgPrxs were increasing in all stages reaching highest level in adult stage. The antibody also showed cross reactivities with corresponding proteins in some cattle helminthes. Natural antibody to FgPrxs could be detected in the sera of mice at 3 and 4 weeks after infection with F. gigantica metacercariae. By immunofluorescence, FgPrxs was highly expressed in tegument and tegumental cells, parenchyma, moderately expressed in cecal epithelial cells in early, juvenile and adult worms. Furthermore, FgPrxs was also detected in the female reproductive organs, including eggs, ovary, vitelline cells, and testis, suggesting that FgPrxs might play an essential role in protecting parasite's tissues from free radical attack during their life cycle. Thus, FgPrxs is one potential candidate for drug therapy and vaccine development. Copyright © 2014 Elsevier Inc. All rights reserved.

Parasites of native and nonnative fishes of the Little Colorado River, Grand Canyon, Arizona

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Choudhury, A.; Hoffnagle, T.L.; Cole, Rebecca A.

2004-01-01

A 2-yr, seasonal, parasitological study of 1,435 fish, belonging to 4 species of native fishes and 7 species of nonnative fishes from the lower Little Colorado River (LCR) and tributary creeks, Grand Canyon, Arizona, yielded 17 species of parasites. These comprised 1 myxozoan (Henneguya exilis), 2 copepods (Ergasilus arthrosis and Lernaea cyprinacea), 1 acarine (Oribatida gen. sp.), 1 piscicolid leech (Myzobdella lugubris), 4 monogeneans (Gyrodactylus hoffmani, Gyrodactylus sp., Dactylogyrus extensus, and Ligictaluridus floridanus), 4 nematodes (Contracaecum sp., Eustrongylides sp., Rhabdochona sp., and Truttaedacnitis truttae), 3 cestodes (Bothriocephalus acheilognathi, Corallobothrium fimbriatum, and Megathylacoides giganteum), and 2 trematodes (Ornithodiplostomum sp. and Posthodiplostomum sp.). Rhabdochona sp. was the only adult parasite native to the LCR. Infection intensities of Ornithodiplostomum sp. and B. acheilognathi were positively correlated with length of the humpback chub Gila cypha. Adult helminths showed a high degree of host specificity, except B. acheilognathi, which was recovered from all fish species examined but was most abundant in cyprinids. Abundance of B. acheilognathi in the humpback chub was highest in the fall and lowest in the summer in both reaches of the LCR. There was no major taxonomic difference in parasite assemblages between the 2 different reaches of the river (LC1 and LC2). Parasite community diversity was very similar in humpback chub, regardless of sampling site or time. The parasite fauna of the LCR is numerically dominated by B. acheilognathi and metacercariae of Ornithodiplostomum sp. The richest and most diverse component community occurred in a nonnative species, the channel catfish Ictalurus punctatus, but infracommunity species richness was highest in a native host, humpback chub.

An Integrated Control Strategy Takes Clonorchis sinensis Under Control in an Endemic Area in South China.

Science.gov (United States)

Huang, Yalan; Huang, Dana; Geng, Yijie; Fang, Shisong; Yang, Fan; Wu, Chunli; Zhang, Hailong; Wang, Miao; Zhang, Ran; Wang, Xin; Wu, Shuang; Cao, Jianping; Zhang, Renli

2017-12-01

Clonorchis sinensis is an important foodborne zoonosis worldwide and prevalent in China for more than 2000 years. According to the experience of controlling Schistosoma japonica, China started to establish the integrated control strategy for C. sinensis in endemic areas. Lou village, the largest village in Shenzhen city in South China was taken as a pilot site. This longitudinal study assessed the infection status of C. sinensis among people and intermediate hosts from 2006 to 2014 in Lou village. After a continuous intervention with the integrated control strategy, the prevalence of C. sinensis decreased significantly to 2.01% in 2014. The infection intensity also reduced significantly with eggs per gram varying from 45.6 ± 3.4 in 2010 to 21.7 ± 1.6 in 2012. There is also a statistically significant decrease of the prevalence of C. sinensis metacercariae in fish hosts from 16.51% in 2008 before the intervention to 5.33% in 2014. All the old-styled toilets were replaced by sanitary ones with a harmless processing design in 2014. No viable parasite eggs were detected in stool samples from the reconstructed toilets. Health education played an important role in changing the eating habits among the local residents, with a significant decrease in the prevalence of eating raw fish from 91.99% in 2008 to 59.87% in 2014. The evaluation suggested that the integrated strategy we have performed in Lou village is effective in controlling the C. sinensis infection and maintaining the infection rate at a lower level, which can be promoted in other endemic areas.

Oral delivery of Bacillus subtilis spore expressing enolase of Clonorchis sinensis in rat model: induce systemic and local mucosal immune responses and has no side effect on liver function.

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Yu, Jinyun; Chen, Tingjin; Xie, Zhizhi; Liang, Pei; Qu, Honglin; Shang, Mei; Mao, Qiang; Ning, Dan; Tang, Zeli; Shi, Mengchen; Zhou, Lina; Huang, Yan; Yu, Xinbing

2015-07-01

Caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensis, human clonorchiasis remains a major public health problem in China. In previous study, we had expressed enolase from C. sinensis (CsENO) on the surface of Bacillus subtilis spore and the recombinant spore induced a pronounced protection in terms of reduced worm burden and eggs per gram feces, suggesting B. subtilis spore as an ideal vehicle for antigen delivery by oral treatment and CsENO as a promising vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgG and IgA levels both in serum and in intestinal mucus from rats orally administrated with B. subtilis spore surface expressing CsENO by ELISA. Lysozyme levels in serum and in intestinal mucus were analyzed too. In addition, IgA-secreting cells in intestine epithelium of the rats were detected by immunohistochemistry assay. The intestinal villi lengths of duodenum, jejunum, and ileum were also measured. Rats orally treated with B. subtilis spore or normal saline were used as controls. Our results showed that, compared with the control groups, oral administration of B. subtilis spore expressing CsENO induced both systemic and local mucosal immune response. The recombinant spores also enhanced non-specific immune response in rats. The spores had no side effect on liver function. Moreover, it might facilitate food utilization and digestion of the rats. Our work will pave the way to clarify the involved mechanisms of protective efficacy elicited by B. subtilis spore expressing CsENO and encourage us to carry out more assessment trails of the oral treated spore to develop vaccine against clonorchiasis.

Infectious bacterial pathogens, parasites and pathological correlations of sewage pollution as an important threat to farmed fishes in Egypt.

Science.gov (United States)

Mahmoud, Mahmoud A; Abdelsalam, Mohamed; Mahdy, Olfat A; El Miniawy, Hala M F; Ahmed, Zakia A M; Osman, Ahmed H; Mohamed, Hussein M H; Khattab, A M; Zaki Ewiss, M A

2016-12-01

This paper is a part of a multi-disciplinary research "Application of Decentralized On-Site Water Treatment System in Egypt for Use in Agriculture and Producing Safe Fish and Animal Proteins". The project aimed to investigate the environmental impact of implementing sewage water before and after treatment using the effluent of the on-site decentralized Japanese' Johkasou system, in agriculture and producing fish protein. The aim is to establish such system in Egypt to strengthen the sanitary conditions of water resources. In the present study, the impact of the sewage pollution in some fish farms at El-Fayyum, Port Said and El-Dakahlia governorates in Egypt was carried out. Water and fish (Oreochromis niloticus and Mugil cephalus) samples were collected from private fish farms of such localities. Bacteriological and chemical examination of water samples revealed the existence of coliforms and many other bacterial species of significant human health hazards. The chemical parameters of water showed a marked deviation from normal levels while examination of fish flesh specimens indicated contamination with Streptococcus Sp., Staphylococcus Sp., and Salmonella in all examined localities. Other bacterial isolates of human health importance (Morganella morganii, Pseudomonas cepacia and Enterococcos durans) were identified. The parasitological examination revealed the presence of encysted metacercariae (EMC); Diplostomatidae, Prohemistomatidae and Heterphyidae. Moreover, two protozoan parasites (Mxyoboulus tilapiae and Ichthyophthirius multifilis) were also recorded. The histopathological examination revealed mild tissue reaction in case of bacterial infection and severe pathological lesions in different organs in case of EMC infection. Lamellar hyperplasia and mononuclear cell infiltration in branchial tissue was common findings. In skeletal muscles, atrophy of muscle fibres, myolysis and myophagia were detected. Copyright © 2016 Elsevier Ltd. All rights reserved.

Molecular and morphometric study of metacercariae and adults of Pseudamphistomum truncatum (Opisthorchiidae) from roach (Rutilus rutilus) and wild American mink (Mustela vison)

DEFF Research Database (Denmark)

Skov, Jakob; Kania, Per Walter; Jørgensen, Thomas Rohde

2008-01-01

Den digene ikte Pseudamphistomum forekommer som metacercarie i skaller fra Furesøen og som adult i mink fra området. Dette er belyst ved morfometriske og molekylære metoder. Fundet er væsentligt, idet parasitten er zoonotisk og kan inficere mennesker, der indtager utilstrækkeligt varmebehandlet f...

Health and condition of endangered young-of-the-year Lost River and Shortnose suckers relative to water quality in Upper Klamath Lake, Oregon, 2014–2015

Science.gov (United States)

Burdick, Summer M.; Conway, Carla M.; Elliott, Diane G.; Hoy, Marshal S.; Dolan-Caret, Amari; Ostberg, Carl O.

2017-10-19

2015; however, hyperplastic and hypertrophic gill tissue and trichodinids on the gills were observed more frequently in 2014. There were more suckers with normal liver color and texture in 2014 than in 2015. The prevalence of suckers with liver inflammation was greater in 2014 and only observed in suckers collected after August 5, whereas liver inflammation occurred intermittently in 2015. Liver glycogen among suckers decreased in late-August 2014 and increased from early August to mid-September 2015. Lost River suckers had greater whole-body triglyceride content but a larger proportion with an absence of visceral fat observed in 2014 than in 2015. In contrast, shortnose suckers were similar between years in regard to both whole-body triglyceride and visceral fat. Black-spot-forming parasites (trematode metacercariae) were observed in a higher prevalence on shortnose suckers but not Lost River suckers in 2014 than in 2015. Opercular deformities were less prevalent in both species in 2014 than in 2015.Neither gross nor histological examination revealed a high prevalence of abnormalities in suckers that clearly indicate a primary mechanism for juvenile mortality in Upper Klamath Lake. Histological abnormalities were almost always focal and minimal or mild except where associated with parasites. Mild to severe focal abnormalities associated with Lernaea sp. attachment sites and encysted digenean (trematode) metacercariae are unlikely to be associated with mortality. Severe and diffuse inflammation and hyperplasia of the gills associated with Ichthyobodo sp. on one Lost River sucker, may indicate a potential cause of mortality. High mortality may have primarily occurred outside our study period (for example, in spring or over winter), or was caused by a factor that could not be detected with our methods (for example, predation). Alternatively, abnormalities in a small percentage of passively captured suckers in Upper Klamath Lake may indicate health-related issues that

Acquired and introduced macroparasites of the invasive Cuban treefrog, Osteopilus septentrionalis

Directory of Open Access Journals (Sweden)

Nicole Ortega

2015-12-01

Full Text Available Because shifts in host–parasite relationships can alter host populations, attention should be given to the parasites that introduced species take with them or acquire in their introduced range. The Cuban treefrog, Osteopilus septentrionalis, is a successful invasive species in Florida with its parasites in the native range being well-documented, but there is a void in the literature regarding what parasites were lost or introduced in its expansion. We necropsied 330 O. septentrionalis from Tampa, FL and compared their macroparasites to those of O. septentrionalis in their native range and to the parasites of anurans native to the Tampa, FL area to determine the species O. septentrionalis likely introduced or acquired in Florida. At least nine parasite species (Aplectana sp., Oswaldocruzia lenteixeirai, Cylindrotaenia americana, Physaloptera sp., Rhabdias sp., Centrorhynchus sp., unidentified trematode metacercariae, unidentified larval acuariids, and unidentified pentastomids were isolated. We found no differences in parasite communities of adult male and female frogs, which averaged 19.36 parasite individuals and 1.39 parasite species per adult frog, and had an overall prevalence of 77.52%. Acuariid larvae were likely acquired by O. septentrionalis in FL because they are not found in their native range. O. lenteixeirai was likely introduced because it is commonly reported in O. septentrionalis' native range but has never been reported in FL-native anurans. Aplectana sp. is also likely introduced because it has been reported in several anurans in Cuba but only reported once in Florida. O. septentrionalis tended to harbor fewer of its native parasites in the introduced range, which is consistent with the enemy release hypothesis and potentially creates an immunological advantage for this invasive host. Because native populations can be threatened by introduced parasites, there is a need to further explore the frequency and rate at which

Effect of the Filamentous Fungus Mucor circinelloides On The Development of Eggs of the Rumen Fluke Calicophoron daubneyi (Paramphistomidae).

Science.gov (United States)

Arroyo, Fabián; Hernández, José A; Cazapal-Monteiro, Cristiana F; Pedreira, José; Sanchís, Jaime; Romasanta, Ángel; Sánchez-Andrade, Rita; Paz-Silva, Adolfo; Arias, María S

2017-06-01

Ruminants infected by Paramphistomidae flukes shed eggs in the feces, which pass through different stages in the environment until the infective stages (metacercariae) are reached. The activity of the soil fungus Mucor circinelloides on the development of eggs of the rumen fluke Calicophoron daubneyi was presently tested with 3 probes, i.e., in petri plates, feces, and an aqueous environment (tubes). The effect of the fungus was assessed by recording the numbers of undeveloped, nonviable, and embryonated eggs. Nonviable eggs were considered when vacuolization occurred, the inner structures were not clearly observed, the eggshell was broken, or the embryo inside was destroyed. By considering the ability of hyphae of M. circinelloides to develop in the presence of C. daubneyi eggs, attach to their surface, and penetrate and destroy the inner embryo, this ovicidal effect was classified as type 3. After a period of 50 days, the percentage of undeveloped eggs in the feces of infected cattle was 40%; furthermore, 27% of the eggs were nonviable, and 33% were embryonated (1 miracidium inside). The addition of 4 doses of M. circinelloides spores directly onto the feces resulted in 9-31% undeveloped eggs, 38-60% nonviable eggs, and 9-21% embryonated eggs, and no statistical significances were obtained among the different doses. Placing the eggs of C. daubneyi into an aqueous solution containing 10 7 spores of M. circinelloides/ml for 29 days resulted in 43% undeveloped eggs, 40% nonviable eggs, and 17% embryonated eggs, whereas in the controls, the percentages were 48%, 12%, and 40%, respectively. These data demonstrate the usefulness of the spores of the fungus M. circinelloides in limiting the development of the eggs of the trematode C. daubneyi.

Time-dependent tegumental surface changes in juvenile Fasciola gigantica in response to triclabendazole treatment in goat.

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Shareef, P A Ahammed; Brennan, Gerard P; McVeigh, Paul; Khan, M A Hannan; Morphew, Russell M; Mousley, Angela; Marks, Nikki J; Saifullah, M K; Brophy, Peter M; Maule, Aaron G; Abidi, S M A

2014-08-01

Triclabendazole (TCBZ), the anthelmintic drug active against both mature and immature liver flukes, was used to investigate the effect of in vivo treatment on the tegumental surface of juvenile Fasciola gigantica. Five goats were infected with 150 F. gigantica metacercariae each by oral gavage. Four of them were treated with single dose of TCBZ at 10mg/kg at four weeks post-infection. They were euthanized at 0 (untreated), 24, 48, 72 and 96h post treatment. Juvenile flukes were manually retrieved from the goat livers and processed for scanning electron microscopy. In control flukes, the anterior region was adorned with sharply pointed spines projecting away from the surface, while in the posterior region, spines become shorter and narrower, loosing serration and with the appearance of distinct furrows and papillae. The dorsal surface retained the same pattern of surface architecture similar to that of ventral surface. Flukes obtained from 24h post-treatment did not show any apparent change and were still very active. However, there were limited movements and some blebbing, swelling, deposition of tegumental secretions and some flattening displayed by the flukes of 48h post-treatment. All the worms were found dead 72h post-treatment and showed advanced level of tegumental disruptions, consisting of severe distortion of spines, sloughing off the tegument to expose the basal lamina, formation of pores and isolated patches of lesions. By 96h post-treatment, the disruption was extremely severe and the tegument was completely sheared off causing deeper lesions that exposed the underlying musculature. The disruption was more severe at posterior than anterior region and on ventral than dorsal surface. The present study further establishes the time-course of TCBZ action in vivo with 100% efficacy against the juvenile tropical liver fluke. Copyright © 2014 Elsevier B.V. All rights reserved.

Metabolic profiling of an Echinostoma caproni infection in the mouse for biomarker discovery.

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Jasmina Saric

Full Text Available BACKGROUND: Metabolic profiling holds promise with regard to deepening our understanding of infection biology and disease states. The objectives of our study were to assess the global metabolic responses to an Echinostoma caproni infection in the mouse, and to compare the biomarkers extracted from different biofluids (plasma, stool, and urine in terms of characterizing acute and chronic stages of this intestinal fluke infection. METHODOLOGY/PRINCIPAL FINDINGS: Twelve female NMRI mice were infected with 30 E. caproni metacercariae each. Plasma, stool, and urine samples were collected at 7 time points up to day 33 post-infection. Samples were also obtained from non-infected control mice at the same time points and measured using (1H nuclear magnetic resonance (NMR spectroscopy. Spectral data were subjected to multivariate statistical analyses. In plasma and urine, an altered metabolic profile was already evident 1 day post-infection, characterized by reduced levels of plasma choline, acetate, formate, and lactate, coupled with increased levels of plasma glucose, and relatively lower concentrations of urinary creatine. The main changes in the urine metabolic profile started at day 8 post-infection, characterized by increased relative concentrations of trimethylamine and phenylacetylglycine and lower levels of 2-ketoisocaproate and showed differentiation over the course of the infection. CONCLUSION/SIGNIFICANCE: The current investigation is part of a broader NMR-based metabonomics profiling strategy and confirms the utility of this approach for biomarker discovery. In the case of E. caproni, a diagnosis based on all three biofluids would deliver the most comprehensive fingerprint of an infection. For practical purposes, however, future diagnosis might aim at a single biofluid, in which case urine would be chosen for further investigation, based on quantity of biomarkers, ease of sampling, and the degree of differentiation from the non

Use of fish parasite species richness indices in analyzing anthropogenically impacted coastal marine ecosystems

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Dzikowski, R.; Paperna, I.; Diamant, A.

2003-10-01

The diversity of fish parasite life history strategies makes these species sensitive bioindicators of aquatic ecosystem health. While monoxenous (single-host) species may persist in highly perturbed, extreme environments, this is not necessarily true for heteroxenous (multiple-host) species. As many parasites possess complex life cycles and are transmitted through a chain of host species, their dependency on the latter to complete their life cycles renders them sensitive to perturbed environments. In the present study, parasite communities of grey mullet Liza aurata and Liza ramada (Mugilidae) were investigated at two Mediterranean coastal sites in northern Israel: the highly polluted Kishon Harbor (KH) and the relatively unspoiled reference site, Ma'agan Michael (MM). Both are estuarine sites in which grey mullet are one of the most common fish species. The results indicate that fish at the polluted site had significantly less trematode metacercariae than fish at the reference site. Heteroxenous gut helminths were completely absent at the polluted sampling site. Consequently, KH fish displayed lower mean parasite species richness. At the same time, KH fish mean monoxenous parasite richness was higher, although the prevalence of different monoxenous taxa was variable. Copepods had an increased prevalence while monogenean prevalence was significantly reduced at the polluted site. This variability may be attributed to the differential susceptibility of the parasites to the toxicity of different pollutants, their concentration, the exposure time and possible synergistic effects. In this study, we used the cumulative species curve model that extrapolates "true" species richness of a given habitat as a function of increasing sample size. We considered the heteroxenous and monoxenous species separately for each site, and comparison of curves yielded significant results. It is proposed to employ this approach, originally developed for estimating the "true" parasite

Theletrum lamothei sp. nov. (Digenea, parasite of Echidna nocturna from Cuajiniquil, Guanacaste, and other digenes of marine fishes from Costa Rica

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Gerardo Pérez-Ponce de León

1998-06-01

Full Text Available A new species of Theletrum is described from the intestine of two palenose morays, Echidna nocturna, collected in Cuajiniquil, Guanacaste Province, Costa Rica. The new species differs from the type species, T. fustiforme Linton, 1910 by having a subspherical pars prostatica, a subspherical seminal vesicle extending anteriorly to the anterior border of the acetabulum, by the presence of a poorly developed hermaphroditic sac, and by having a larger body size. We also report eight additional species of digeneans parasitizing marine fishes in several localities along the Atlantic and Pacific coast of Costa Rica: Bianium simonei, Didymozoinae (metacercariae, Ectenurus virgulus, Hypocreadium myohelicatum, Lecithochirium microstomum, Pseudolecithaster sp., Stephanostomum casum, and Tergestia laticollis. In addition, we present an updated list of helminth parasites of marine fish from Costa Rica and discuss the importance of including parasites as an integral part of biodiversity inventories.Se describe una especie nueva de digéneo del género Theletrum, parásito del intestino de una "morena pecosa", Echidna nocturna, recolectada en Playa Cuajiniquil, Provincia de Guanacaste, Costa Rica. La nueva especie es diferente de T. fustiforme Linton, 1910, la especie tipo, por presentar una pars prostatica subesférica, una vesícula seminal subesférica que se extiende hasta el borde anterior del acetábulo, por la presencia de un saco hermafrodita y por una mayor longitud del cuerpo. También se registran ocho especies adicionales de digéneos parásitos de peces marinos en varias localidades de las costas del Atlántico y del Pacífico de Costa Rica. Por último, presentamos una lista actualizada de los helmintos parásitos de peces marinos y dulceacuícolas de Costa Rica y discutimos la importancia de incluir a los parásitos como una parte integral de las investigaciones sobre biodiversidad.

Relação entre fator de condição relativo (Kn e abundância de ectoparasitos de brânquias, em duas espécies de ciclídeos da bacia do rio Paraná, Brasil = Relation between the relative condition factor (Kn and the abundance of gill ectoparasites in two species of cichlids from the Paraná river basin, Brazil

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Fábio Hideki Yamada

2008-04-01

Full Text Available Foram analisados 33 espécimes de Satanoperca pappaterra, capturados entre março de 2004 e junho de 2005, na planície de inundação do alto rio Paraná e 33 espécimes de Crenicichla niederleinii, capturados entre novembro de 2005 e novembro de 2006, no reservatório de Itaipu. Ambos estavam parasitados por monogenéticos, pertencentes ao gênero Sciadicleithrum e por metacercárias de Ascocotyle sp. O fator de condição relativo (Kn não diferiu significativamente entre indivíduos parasitados e não-parasitados das duas espécies de hospedeiros. Apenas Sciadicleithrum sp.1 parasito de S. pappaterra apresentou correlação positiva e significativa entre o fator de condição relativo (Kn e a abundância de parasitismo. Por outro lado, Ascocotyle sp. de ambos os hospedeiros e Sciadicleithrum sp.2parasito de C. niederleinii não apresentaram correlações entre o Kn e a abundância de parasitismo.The study analyzed 33 specimens of Satanoperca pappaterra, captured from the Upper Paraná River floodplain between March 2004 and June of 2005, and 33 specimens of Crenicichla niederleinii captured from the Itaipu Reservoir between November 2005 and November 2006. Both were parasitized by monogeneans pertaining to the genus Sciadicleithrum and by metacercariae of Ascocotyle sp. The relative condition factor (Kn did not differ significantly between parasitized and unparasitized individuals from the two host species. Only Sciadicleithrum sp.1, parasite of S. pappaterra, presented positive and significant correlation between the relative condition factor (Kn and the abundance of parasitism. On the other hand, Ascocotyle sp. in both hosts and Sciadicleithrum sp.2 parasite of C. niederleiniidid not present correlations between the Kn and the abundance of parasitism.

Coproantígenos de Fasciola hepatica de posible utilidad en el diagnóstico de la fascioliasis

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Ana M. Espino

2000-04-01

Full Text Available En el presente estudio se describen cuáles son los antígenos de Fasciola hepatica que están presentes en las heces de pacientes con fascioliasis crónica y de ratas infectadas experimentalmente con metacercarias de F. hepatica. Mediante el empleo de la técnica de inmunoelectrotransferencia (Western blot con un suero hiperinmune obtenido frente a antígenos de excreción/secreción de adultos de F. hepatica, se pudo demostrar que en las heces de los pacientes se encuentran antígenos de posible interés para el diagnóstico con masas moleculares de 14, 19, 20, 23, 25, 32, 46, 51 y 62 kilodaltons (kDa. Además, se comprobó que algunos de estos péptidos (los de 14, 20, 23 y 51 kDa son reconocidos también por la mayoría de los sueros de pacientes crónicos. Después de purificar por cromatografía de afinidad los antígenos presentes en heces de ratas con 6 a 12 semanas de infección, empleando para ello el anticuerpo monoclonal ES78 acoplado a Sepharosa 4B-CNBr, se identificaron seis polipéptidos de 11, 14, 26, 32, 47 y 51 kDa; otros tres polipéptidos, de 17, 24 y 66 kDa, solo se pudieron identificar en heces de ratas con 10 a 12 semanas de infección. Nuestros resultados sugieren que estos polipéptidos pueden ser antígenos comunes a ambos estadios parasitarios y constituir importantes marcadores de la fascioliasis aguda y crónica, en particular los de 14, 24, 26 y 51 kDa, puesto que reaccionaron con el inmunosuero, los sueros humanos y el anticuerpo monoclonal ES78.

Alteration in Haematological and Liver Function Indices during Human Infection with Fasciola spp. Post Treatment with Triclabendazole

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M.I. Edalatzadeh

2006-07-01

Full Text Available Introduction & Objective: Fascioliasis is a zoonotic parasitic disease, caused by the liver fluke, Fasciola spp.. Human is occasional host when ingesting the metacercaria by eating contaminated aquatic vegetable. In the two past decades, human fasciolasis was emerging as a problem of public health in the Guilan province; in Anzali city. Triclabendazole is a novel anti-helmenthic that during recent years has been used for fascioliasis treatment in this region. The aim of the present work is to study alteration in haematological and liver function indices during human infection with Fasciola spp. pre and post treatment with triclabendazoleMaterials & Methods: The present work is a longitudinal clinical trail. In this regard, fifty confirmed fasciolasis patients, were chosen for parasitological, hematological and biochemical examinations pre-therapy as well as 1 and 6 months post-therapy. Formalin-ether and modified Telemann methods were used for stool examination. For Fasciola antibody detection ELISA technique was employed. Hematological and biochemical tests were performed by standard methods. Results: Results indicated that, triclabendazole efficacy was 74% after usage as one dose of 20mg/kg and reached to 88% after repeating in the next month. Before triclabebdazole therapy the Hb and HCT of the patients were slightly found lower than normal ranges, meanwhile the ESR and eosinophil percentages were higher. However following receiving the drug, in the cured individuals, the indices returned to the normal ranges but in the non-cured individuals were not shifted to the normal. On the other hand liver function indices of the patients mostly were at normal ranges before and following drug therapy.Conclusion: In conclusion haematological indices could be valuable indicator for successful therapy of patients treated with triclabendazole.

Field survey focused on Opisthorchis viverrini infection in five provinces of Cambodia.

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Miyamoto, Kazuko; Kirinoki, Masashi; Matsuda, Hajime; Hayashi, Naoko; Chigusa, Yuichi; Sinuon, Muth; Chuor, Char Meng; Kitikoon, Viroj

2014-04-01

Opisthorchiasis is endemic in Thailand and Lao People's Democratic Republic and constitutes a major public health problem throughout the Mekong Basin. Although Cambodia is located in the Mekong Basin, the status of O. viverrini infection in that country was not previously clarified. This research was conducted to document the extent and distribution of O. viverrini infection in Cambodia. Surveillance was conducted in 55 villages in five Cambodian provinces. Research tools included stool examination using the Kato-Katz thick-smear technique, identification of intermediate hosts, and interviews covering factors related to O. viverrini infection. Some larvae and egg-positive stool samples were examined using PCR to detect O. viverrini DNA. A total of 16,082 stool samples from the 55 villages were examined, of which 1232 were egg positive. In 15 villages with egg-positive rates of greater than 10%, eggs were found in 998 of 3585 stool samples, for an egg-positive rate of 27.8%. PCR analysis showed that 30 of 33 samples were positive for O. viverrini DNA from five villages in Kampong Cham and Kampong Thom provinces. The first intermediate host Bithynia siamensis siamensis was identified in the target areas of Takaev, Kandal, and Kampong Cham provinces. Cercariae were identified morphologically as O. viverrini and some were confirmed using PCR. Metacercariae of O. viverrini were identified by morphologic observations, animal experiments, or PCR in six species of fish in the target areas. Four Cambodian provinces were identified as endemic areas of O. viverrini infection. Careful planning is necessary for effective field surveys, because complex environmental factors might be involved in the distribution of O. viverrini infection-endemic areas in Cambodia. Many problems remain to be resolved regarding the status of O. viverrini infection in Cambodia, and a nationwide baseline survey is necessary. Copyright © 2013 Elsevier B.V. All rights reserved.

The Regulatory Roles of Toll-Like Receptor 4 in Secretions of Type 1/Type 2 Relative Cytokines by Splenocytes and Dendritic Cells Exposed to Clonorchis sinensis Excretory/Secretory Products.

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Hua, Hui; Du, Ying; Ma, Rui; Zhang, Bei-Bei; Yu, Qian; Li, Bo; Xu, Jiang-Tao; Li, Xiang-Yang; Tang, Ren-Xian; Yan, Chao; Zheng, Kui-Yang

2018-02-01

The roles of TLR4 in mediation of innate immune response and in regulation of adaptive immune responses triggered by Clonorchis sinensis remain unknown. In the present study, splenocytes derived from C3H/HeN (TLR4 wild ) and C3H/Hej mice (TLR4 mut ) that were infected with 45 metacercariae of C. sinensis were harvested, then stimulated by C. sinensis excretory/secretory products (ESP) or medium (control) for 48 h, respectively. Meanwhile, bone marrow-derived dendritic cells (BMDCs) from normal C3H/HeN and C3H/Hej mice were prepared and stimulated with medium, ESP, LPS, or ESP+LPS for 24 h, respectively. The supernatants were collected, and the concentrations of type 1 and type 2 relative cytokines were determined by ELISA. The maturation of BMDCs indicated by surface markers of CD80, CD86, and MHC II was evaluated by flow cytometry. The results showed that the levels of IFN-γ, IL-6, TNF-α, and IL-10 in the splenocytes from C. sinensis-infected TLR4 mut mice were significantly lower than those from TLR4 wild mice when they were further exposed to ESP. For BMDCs, the productions of the cytokines IL-12p70 and IL-10, but not IL-4, in the BMDCs from TLR4 mutation mice were predominantly decreased compared with those from TLR4 wild mice when the BMDCs were co-stimulated by ESP combined with LPS. Flow cytometry analysis showed that ESP could significantly decrease the high levels of CD80, CD86, and MHC II which were elevated by LPS. In conclusion, these data suggest that TLR4 may play a regulatory role in type 1 immune responses during C. sinensis infection.

Molecular characterization of an α-N-acetylgalactosaminidase from Clonorchis sinensis.

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Lee, Myoung-Ro; Yoo, Won Gi; Kim, Yu-Jung; Kim, Dae-Won; Cho, Shin-Hyeong; Hwang, Kwang Yeon; Ju, Jung-Won; Lee, Won-Ja

2012-11-01

The α-N-acetylgalactosaminidase (α-NAGAL) is an exoglycosidase that selectively cleaves terminal α-linked N-acetylgalactosamines from a variety of sugar chains. A complementary DNA (cDNA) clone encoding a novel Clonorchis sinensis α-NAGAL (Cs-α-NAGAL) was identified in the expressed sequence tags database of the adult C. sinensis liver fluke. The complete coding sequence was 1,308 bp long and encoded a 436-residue protein. The selected glycosidase was manually curated as α-NAGAL (EC 3.2.1.49) based on a composite bioinformatics analysis including a search for orthologues, comparative structure modeling, and the generation of a phylogenetic tree. One orthologue of Cs-α-NAGAL was the Rattus norvegicus α-NAGAL (accession number: NP_001012120) that does not exist in C. sinensis. Cs-α-NAGAL belongs to the GH27 family and the GH-D clan. A phylogenetic analysis revealed that the GH27 family of Cs-α-NAGAL was distinct from GH31 and GH36 within the GH-D clan. The putative 3D structure of Cs-α-NAGAL was built using SWISS-MODEL with a Gallus gallus α-NAGAL template (PDB code 1ktb chain A); this model demonstrated the superimposition of a TIM barrel fold (α/β) structure and substrate binding pocket. Cs-α-NAGAL transcripts were detected in the adult worm and egg cDNA libraries of C. sinensis but not in the metacercaria. Recombinant Cs-α-NAGAL (rCs-α-NAGAL) was expressed in Escherichia coli, and the purified rCs-α-NAGAL was recognized specifically by the C. sinensis-infected human sera. This is the first report of an α-NAGAL protein in the Trematode class, suggesting that it is a potential diagnostic or vaccine candidate with strong antigenicity.

Increased hepatic Th2 and Treg subsets are associated with biliary fibrosis in different strains of mice caused by Clonorchis sinensis.

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Bei-Bei Zhang

Full Text Available Previous studies showed that CD4+T cells responses might be involved in the process of biliary fibrosis. However, the underlying mechanism resulting in biliary fibrosis caused by Clonorchis sinensis remains not yet fully elucidated. The objectives of the present study were to investigate the different profiles of hepatic CD4+T cell subsets (Th1, Th2, Th17 and Treg cells and their possible roles in the biliary fibrosis of different strains of mice (C57BL/6, BALB/c and FVB mice induced by C. sinensis infection. C57BL/6, BALB/c and FVB mice were orally gavaged with 45 metacercariae. All mice were sacrificed on 28 days post infection in deep anesthesia conditions. The leukocytes in the liver were separated to examine CD4+T cell subsets by flow cytometry and the left lobe of liver was used to observe pathological changes, collagen depositions and the concentrations of hydroxyproline. The most serious cystic and fibrotic changes appeared in FVB infected mice indicated by gross observation, Masson's trichrome staining and hydroxyproline content detection. In contrast to C57BL/6 infected mice, diffuse nodules and more intensive fibrosis were observed in the BALB/c infected mice. No differences of the hepatic Th1 subset and Th17 subset were found among the three strains, but the hepatic Th2 and Treg cells and their relative cytokines were dramatically increased in the BALB/c and FVB infected groups compared with the C57BL/6 infected group (P<0.01. Importantly, increased Th2 subset and Treg subset all positively correlated with hydroxyproline contents (P<0.01. This result for the first time implied that the increased hepatic Th2 and Treg cell subsets were likely to play potential roles in the formation of biliary fibrosis in C. sinensis-infected mice.

Tracing of the Bile-chemotactic migration of juvenile Clonorchis sinensis in rabbits by PET-CT.

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Tae Im Kim

2011-12-01

Full Text Available BACKGROUND: Adult Clonorchis sinensis live in the bile duct and cause clonorchiasis. It is known that the C. sinensis metacercariae excyst in the duodenum and migrate up to the bile duct through the common bile duct. However, no direct evidence is available on the in vivo migration of newly excysted C. sinensis juveniles (CsNEJs. Advanced imaging technologies now allow the in vivo migration and localization to be visualized. In the present study, we sought to determine how sensitively CsNEJs respond to bile and how fast they migrate to the intrahepatic bile duct using PET-CT. METHODOLOGY/PRINCIPAL FINDINGS: CsNEJs were radiolabeled with (18F-fluorodeoxyglucose ((18F-FDG. Rabbits with a gallbladder contraction response to cholecystokinin-8 (CCK-8 injection were pre-screened using cholescintigraphy. In these rabbits, gallbladders contracted by 50% in volume at an average of 11.5 min post-injection. The four rabbits examined were kept anesthetized and a catheter inserted into the mid duodenum. Gallbladder contraction was stimulated by injecting CCK-8 (20 ng/kg every minute over the experiment. Anatomical images were acquired by CT initially and dynamic PET was then carried out for 90 min with a 3-min acquisition per frame. Twelve minutes after CCK-8 injection, about 3,000 (18F-FDG-labeled CsNEJs were inoculated into the mid duodenum through the catheter. Photon signals were detected in the liver 7-9 min after CsNEJs inoculation, and these then increased in the whole liver with stronger intensity in the central area, presenting that the CsNEJs were arriving at the intrahepatic bile ducts. CONCLUSION: In the duodenum, CsNEJs immediately sense bile and migrate quickly with bile-chemotaxis to reach the intrahepatic bile ducts by way of the ampulla of Vater.

Immune response induced by oral delivery of Bacillus subtilis spores expressing enolase of Clonorchis sinensis in grass carps (Ctenopharyngodon idellus).

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Jiang, Hongye; Chen, Tingjin; Sun, Hengchang; Tang, Zeli; Yu, Jinyun; Lin, Zhipeng; Ren, Pengli; Zhou, Xinyi; Huang, Yan; Li, Xuerong; Yu, Xinbing

2017-01-01

Clonorchiasis, caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensisis (C.sinensis), remains a common public health problem. New effective prevention strategies are still urgent to control this food-borne infectious disease. The previous studies suggested Bacillus subtilis (B. subtilis) spores was an ideal vaccines delivery system, and the C.sinensis enolase (CsENO) was a potential vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgM levels by ELISA in sera, intestinal mucus and skin mucus in grass carps (Ctenopharyngodon idella) through oral administration with B. subtilis spores surface expressing CsENO. In addition, immune-related genes expression was also measured by qRT-PCR. Grass carps orally treated with B. subtilis spores or normal forages were used as controls. The results of ELISA manifested that specific IgM levels of grass carps in CsENO group in sera, intestine mucus and skin mucus almost significantly increased from week 4 post the first oral administration when compared to the two control groups. The levels of specific IgM reached its peak in intestine mucus firstly, then in sera, and last in skin mucus. qRT-PCR results showed that 5 immune-related genes expression had different degree of rising trend in CsENO group when compared to the two control groups. Our study demonstrated that orally administrated with B. subtilis spores expressing CsENO induced innate and adaptive immunity, systemic and local mucosal immunity, and humoral and cellular immunity. Our work may pave the way to clarify the exact mechanisms of protective efficacy elicited by B. subtilis spores expressing CsENO and provide new ideas for vaccine development against C. sinensis infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

Molecular cloning and characterization of taurocyamine kinase from Clonorchis sinensis: a candidate chemotherapeutic target.

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Jing-Ying Xiao

2013-11-01

Full Text Available BACKGROUND: Adult Clonorchis sinensis lives in the bile duct and causes endemic clonorchiasis in East Asian countries. Phosphagen kinases (PK constitute a highly conserved family of enzymes, which play a role in ATP buffering in cells, and are potential targets for chemotherapeutic agents, since variants of PK are found only in invertebrate animals, including helminthic parasites. This work is conducted to characterize a PK from C. sinensis and to address further investigation for future drug development. METHODOLOGY/PRINCIPAL FINDINGS: [corrected] A cDNA clone encoding a putative polypeptide of 717 amino acids was retrieved from a C. sinensis transcriptome. This polypeptide was homologous to taurocyamine kinase (TK of the invertebrate animals and consisted of two contiguous domains. C. sinensis TK (CsTK gene was reported and found consist of 13 exons intercalated with 12 introns. This suggested an evolutionary pathway originating from an arginine kinase gene group, and distinguished annelid TK from the general CK phylogenetic group. CsTK was found not to have a homologous counterpart in sequences analysis of its mammalian hosts from public databases. Individual domains of CsTK, as well as the whole two-domain enzyme, showed enzymatic activity and specificity toward taurocyamine substrate. Of the CsTK residues, R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 were found to participate in binding taurocyamine. CsTK expression was distributed in locomotive and reproductive organs of adult C. sinensis. Developmentally, CsTK was stably expressed in both the adult and metacercariae stages. Recombinant CsTK protein was found to have low sensitivity and specificity toward C. sinensis and platyhelminth-infected human sera on ELISA. CONCLUSION: CsTK is a promising anti-C. sinensis drug target since the enzyme is found only in the C. sinensis and has a substrate specificity for taurocyamine, which is different from its mammalian counterpart

Vaccine potential of recombinant cathepsinL1G against Fasciola gigantica in mice.

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Changklungmoa, Narin; Phoinok, Natthacha; Yencham, Chonthicha; Sobhon, Prasert; Kueakhai, Pornanan

2016-08-15

In this study, we characterized and investigated the vaccine potential of FgCatL1G against Fasciola gigantica infection in mice. Recombinant mature FgCatL1G (rmFgCatL1G) was expressed in Escherichia coli BL21. The vaccination was performed in Imprinting Control Region (ICR) mice (n=10) by subcutaneous injection with 50μg of rmFgCatL1G combined with Freund's adjuvant. Two weeks after the second boost, mice were infected with 15 metacercariae by the oral route. The percents of protection of rmFgCatL1G vaccine were estimated to be 56.5% and 58.3% when compared with non vaccinated-infected and adjuvant-infected controls, respectively. Antibodies in the immune sera of vaccinated mice were shown by immunoblot to react with the native FgCatL1s in the extract of all stages of parasites and rmFgCatL1H, recombinant pro - FgCatL1 (rpFgCatL1). By immunohistochemistry, the immune sera also reacted with FgCatL1s in the caecal epithelial cells of the parasites. The levels of IgG1 and IgG2a in the immune sera, which are indicative of Th2 and Th1 immune responses, were also increased with IgG1 predominating. The levels of serum glutamic oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) in rmFgCatL1G-immunized group showed no significant difference from the control groups, but pathological lesions of livers in rmFgCatL1G-immunized group showed significant decrease when compared to the control groups. This study indicates that rmFgCatL1G has a vaccine potential against F. gigantica in mice, and this potential will be tested in larger livestock animals. Copyright © 2016 Elsevier B.V. All rights reserved.

Parasitological survey of mangrove oyster, Crassostrea rhizophorae, in the Pacoti River Estuary, Ceará State, Brazil.

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Sabry, Rachel Costa; Gesteira, Tereza Cristina Vasconcelos; Magalhães, Aimê Rachel Magenta; Barracco, Margherita Anna; Guertler, Cristhiane; Ferreira, Liana Pinho; Vianna, Rogério Tubino; da Silva, Patrícia Mirella

2013-01-01

The mangrove oyster, Crassostrea rhizophorae (Bivalvia, Ostreidae) is commonly collected by fisherwomen in the estuaries of the Ceará State (CE), Northeastern Brazil. Despite the socioeconomic importance of this natural resource, there are few studies on the health of the oysters in this region. This study aimed to survey pathological changes in the mangrove oyster C. rhizophorae in the estuary of the Pacoti River, CE. Adult oysters were collected in August 2008 (N=450) and December 2009 (N=450) at three sites of the Pacoti estuary and in 2010 (N=600) samplings were done quarterly at one site which has showed the higher prevalence de Perkinsus. Macroscopical and histological analyses were used to evaluate pathological changes, Ray's Fluid Thioglycollate Medium (RFTM) to detect Perkinsus spp. and polymerase chain reactions (PCR) and DNA sequencing to identify Perkinsus species. In 2009, RFTM assay detected Perkinsus sp. infecting the tissues of C. rhizophorae with low prevalences of 1.3%, 6.7% e 7.3% in sites 1, 2 and 3, respectively, and in 2010, in site 3, prevalence was 2% (12 of 600 oysters). PCR did not confirm any positive case in 2009 and only 5 in 2010. The phylogenetic analyses strongly indicate that the Perkinsus species infecting oysters C. rhizophorae of this study belongs to Perkinsus beihaiensis. The histology confirmed 11 cases of Perkinsus sp. infecting the C. rhizophorae in 2009, and only two cases in 2010. Nematopsis sp. was the protozoan observed with greater prevalence (up 96.7%). Other found protozoa were: Trichodina, Sphenophrya, Ancistrocoma - like and an unknown ovarian parasite. The metazoa found were the polychaete Polydora with high prevalences, a turbellarian, possibly of the genus Urastoma, an unidentified digenean metacercariae and larvae of cestode Tylocephalum. A continuous monitoring of diseases in bivalves from this natural population is recommended, since the phylogenetic analyses indicate the occurrence of P. beihaiensis

Spatial structure of helminth communities in the golden grey mullet, Liza aurata (Actinopterygii: Mugilidae), from the Western Mediterranean.

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Míguez-Lozano, Raúl; Pardo-Carranza, Trinidad V; Blasco-Costa, Isabel; Balbuena, Juan Antonio

2012-10-01

Ecological investigations regarding the parasite fauna of grey mullets are scarce. The present study provides a detailed description of the helminth communities of Liza aurata in the Spanish western Mediterranean and analyzes the role of spatial, temporal, and host variables in shaping the infracommunities. In total, 204 fish were collected in 2 localities, situated ca. 290 km apart, in spring and fall of 2004 and 2005. A non-metric multidimensional scaling (NMDS) was used to visualize an ordination of the infracommunities according to their relative similarities in parasite abundances. The relationship between infracommunity composition and explanatory variables (host size, locality, year, and season of harvest) was examined by permutational analysis of variance (PERMANOVA) applied to species abundances. Permutational tests for homogeneity of multivariate dispersion were used to test the null hypothesis of no differences in dispersion among groups formed by the factors whose effects were significant in PERMANOVA. A total of 33,241 helminth parasites, belonging to 18 species, was collected, i.e., 12 species of adult digeneans (23% of the parasite specimens), 3 digeneans as metacercariae (68%), 1 acanthocephalan (2.1%), and 2 monogeneans (6.5%). An important part of this helminth fauna is specialized to grey mullets, with a sizable portion of the component community restricted to the Mediterranean and northeast Atlantic. The NMDS ordination indicated high heterogeneity among infrapopulations. However, most differences at both the component and infracommunity level were related to geographic locality. In fact, the PERMANOVA showed that, among the explanatory variables considered, sampling locality accounted for the largest share of variation. The geographical differences observed may be related to local environmental characteristics or to the limited spatial dispersal of the species forming the component community. The latter was supported by the significant portion

A 2.9 kDa Fasciola hepatica-recombinant protein based ELISA test for the detection of current-ovine fasciolosis trickle infected.

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Arias, M; Hillyer, G V; Sánchez-Andrade, R; Suárez, J L; Pedreira, J; Lomba, C; Díaz, P; Morrondo, P; Díez-Baños, P; Paz-Silva, A

2006-04-15

The suitability of an enzyme linked immunosorbent assay (ELISA) test with a 2.9 kDa Fasciola hepatica-recombinant protein (FhrAPS) for diagnosing early and current-ovine fasciolosis was analyzed, and compared to that obtained by using a direct ELISA for detecting F. hepatica-circulating FhES antigens and to the coprological sedimentation for fluke egg quantitation. Fourteen Gallega autochthonous breed sheep were experimentally infected with metacercariae by a trickle system (small repetitive infections) and divided into two groups: G-I represented a primary infection for 34 weeks; G-R, animals with primary infection and reinfected 18 w.a.p.i. Seven sheep were left uninfected as the control group (G-C). Serum IgG antibody values against the FhrAPS rose rapidly by 1st w.a.p.i. in all infected sheep. Antibody levels in those with primary infection (G-I, G-C) peaked at 10 weeks, diminishing slightly and levelling from 16 to 34 weeks. Those with primary infection reinfected at 18 weeks had a rebound effect with the highest values observed. Circulating F. hepatica-ES antigens were detected by the 1st w.a.p.i. in all infected groups peaking at 6 weeks, decreasing rapidly to uninfected control values by 10 weeks of infection. Faecal egg-output started 11 weeks after primary infection. An increase in the IgG antibody as well as antigen responses to the FhrAPS and to anti-FhES from the 18 w.a.p.i. was recorded in G-T and G-R after the challenge infection. Antibody levels remained high whereas antigenemia values diminished after 6 weeks. A positive significant correlation between the IgG response against the FhrAPS and the F. hepatica circulating antigens (r2 = 0.428, p = 0.001) was obtained. In conclusion, our standardized diagnostic ELISA for fasciolosis based on the detection of IgG responses to the FhrAPS would be a valuable tool to diagnosis early and current F. hepatica-infections in sheep.

Vaccine potential of recombinant saposin-like protein 2 against Fasciolosis gigantica in mice.

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Kueakhai, Pornanan; Changklungmoa, Narin; Riengrojpitak, Suda; Chaichanasak, Pannigan; Meemon, Krai; Chaithirayanon, Kulathida; Chantree, Pathanin; Sansri, Veerawat; Itagaki, Tadashi; Sobhon, Prasert

2013-11-12

Saposin-like protein 2 (SAP-2) is a protein that adult of Fasciola spp. use to lyse plasma membrane of red blood cells, so that their contents can be digested by proteases for the parasites' nutrients. Thus SAP-2 is a plausible target for vaccination against these parasites. Recombinant Fasciola gigantica saposin-like protein 2 (rFgSAP-2) was expressed in Escherichia coli BL21 (DE3). A vaccination was performed in ICR mice (n=10) by subcutaneous injection with 50μg of rFgSAP-2 combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 30 F. gigantica metacercariae by oral route. The percentages of protection of rFgSAP-2 vaccine against F. gigantica were estimated to be 76.4-78.5% when compared with non vaccinated-infected and adjuvant-infected controls, respectively. The antibodies in immune sera of vaccinated mice were shown by immuno-blotting to react with native FgSAP-2 in the extract of 2- and 4-week-old juvenile parasites. By determining the levels of IgG1 and IgG2a in the immune sera, which are indicative of Th2 and Th1 immune responses, it was found that both Th1 and Th2 humoral immune response were significantly increased in rFgSAP-2 immunized group compared with the control groups, with higher levels of Th2 (IgG1) than Th1 (IgG2a). The levels of serum aspartate aminotransferase (AST) and alanine transaminase (ALT) in rFgSAP-2-immunized group showed no significant difference from those of the non-immunized and infected group, indicating that early juvenile parasites induced liver parenchyma damage, even though the numbers of worm recoveries were significantly different. This study indicates that rFgSAP-2 has a high potential as a vaccine candidate against F. gigantica in mice, and this potential will be tested in larger economic animals. Copyright © 2013 Elsevier Ltd. All rights reserved.

Application of a coproantigen ELISA as an indicator of efficacy against multiple life stages of Fasciola hepatica infections in sheep.

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George, S D; Vanhoff, K; Baker, K; Lake, L; Rolfe, P F; Seewald, W; Emery, D L

2017-11-15

At present diagnosis of true resistance and determination of drug efficacy in Fasciola hepatica infection rely solely on terminal experiments. The coproantigen ELISA (cELISA) has been reported previously as a sensitive and specific tool appropriate to detect treatment failure, and potentially drug resistance. Two studies were conducted to determine whether the cELISA was appropriate for on-farm efficacy and resistance testing in Australian Merino sheep. In Study 1 sheep were infected orally with 50 F. hepatica metacercariae on three occasions, twelve, six and two weeks prior to a single flukicide treatment with triclabendazole, closantel or albendazole. Sheep were sampled weekly for a further seven weeks prior to necropsy. Following effective treatment, no faecal antigen was detected from 1 week. When immature stages (≤6 weeks) survived treatment, coproantigen reappeared from 6 weeks post-treatment. Therefore, cELISA conducted 1-4 weeks after treatment will demonstrate obvious treatment failure against adult F. hepatica, but is not sufficiently sensitive to detect survival of immature fluke until these reach maturity. In study 2, fluke burdens of sheep necropsied 13 weeks post single infection were compared to fecal worm egg counts (FWEC) and cELISA at necropsy. Regression analysis demonstrated that cELISA correlated strongly with fluke burden, whilst FWEC correlated weakly with cELISA. The correlation between FWEC and fluke burden was also weak, although stronger than that of FWEC with cELISA. The cELISA is an appropriate tool for monitoring effectiveness of treatments against Fasciola hepatica if an adult infection is present, however when immature stages of the parasite are present it is not as reliable. Where immature parasites are present it is recommended that initial cELISA be followed with a secondary cELISA at least 6 weeks after treatment to ensure resistance to immature stages is detected. Further testing is justified for monitoring the effectiveness

Low virulence potential and in vivo transformation ability in the honey bee venom treated Clinostomum complanatum.

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Rehman, Abdur; Ullah, Rizwan; Jaiswal, Neeshma; Khan, M A Hannan; Rehman, Lubna; Beg, Mirza Ahmar; Malhotra, Sandeep K; Abidi, S M A

2017-12-01

The helminth parasites possess great capabilities to adapt themselves within their hosts and also develop strategies to render the commonly used anthelmintics ineffective leading to the development of resistance against these drugs. Besides using anthelmintics the natural products have also been tested for their anti-parasitic effects. Therapeutic efficacy of honey bee venom (HBV) has been tested in various ailments including some protozoal infections but very little is known about its anthelmintic properties. To investigate the anthelmintic effect of HBV the excysted progenetic metacercariae of Clinostomum complanatum, a heamophagic, digenetic trematode with zoonotic potential, infecting a wide variety of hosts, were obtained from Trichogaster fasciatus, a forage fish, which serves as the intermediate host. The metacercarial worms were in vitro incubated in RPMI-1640 medium containing HBV along with the controls which were devoid of HBV for the analysis of worm motility, enzyme activity, polypeptide profile and surface topographical changes. The motility of the worms was significantly reduced in a time dependent manner with an increase in the concentration of HBV. Following incubation of worms the release of cysteine proteases was inhibited in the presence of HBV as revealed by gelatine substrate gel zymography. As well as the polypeptide profile was also significantly influenced, particularly intensity/expression of M r 19.4 kDa, 24 kDa and 34 kDa was significantly reduced upon HBV treatment. The HBV treatment also inhibited antioxidant enzyme, superoxide dismutase (SOD) and Glutathione-S-transferase (GST) significantly (p < 0.05) in the worms. The scanning electron microscopy of the HBV treated worms revealed tegumental disruptions and erosion of papillae as well as spines showing vacuolation in the tegument. The HBV treated worms also showed a marked decline in the transformation rate when introduced into an experimental host which further reflect the

Molecular Cloning and Characterization of Taurocyamine Kinase from Clonorchis sinensis: A Candidate Chemotherapeutic Target

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Tokuhiro, Shinji; Nagataki, Mitsuru; Jarilla, Blanca R.; Nomura, Haruka; Kim, Tae Im; Hong, Sung-Jong; Agatsuma, Takeshi

2013-01-01

Background Adult Clonorchis sinensis lives in the bile duct and causes endemic clonorchiasis in East Asian countries. Phosphagen kinases (PK) constitute a highly conserved family of enzymes, which play a role in ATP buffering in cells, and are potential targets for chemotherapeutic agents, since variants of PK are found only in invertebrate animals, including helminthic parasites. This work is conducted to characterize a PK from C. sinensis and to address further investigation for future drug development. Methology/Principal findings A cDNA clone encoding a putative polypeptide of 717 amino acids was retrieved from a C. sinensis transcriptome. This polypeptide was homologous to taurocyamine kinase (TK) of the invertebrate animals and consisted of two contiguous domains. C. sinensis TK (CsTK) gene was reported and found consist of 13 exons intercalated with 12 introns. This suggested an evolutionary pathway originating from an arginine kinase gene group, and distinguished annelid TK from the general CK phylogenetic group. CsTK was found not to have a homologous counterpart in sequences analysis of its mammalian hosts from public databases. Individual domains of CsTK, as well as the whole two-domain enzyme, showed enzymatic activity and specificity toward taurocyamine substrate. Of the CsTK residues, R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 were found to participate in binding taurocyamine. CsTK expression was distributed in locomotive and reproductive organs of adult C. sinensis. Developmentally, CsTK was stably expressed in both the adult and metacercariae stages. Recombinant CsTK protein was found to have low sensitivity and specificity toward C. sinensis and platyhelminth-infected human sera on ELISA. Conclusion CsTK is a promising anti-C. sinensis drug target since the enzyme is found only in the C. sinensis and has a substrate specificity for taurocyamine, which is different from its mammalian counterpart, creatine. PMID:24278491

Molecular and structural characteristics of multidrug resistance-associated protein 7 in Chinese liver fluke Clonorchis sinensis.

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Dai, Fuhong; Yoo, Won Gi; Lee, Ji-Yun; Lu, Yanyan; Pak, Jhang Ho; Sohn, Woon-Mok; Hong, Sung-Jong

2017-03-01

Multidrug resistance-associated protein 7 (MRP7, ABCC10) is a C subfamily member of the ATP-binding cassette (ABC) superfamily. MRP7 is a lipophilic anion transporter that pumps endogenous and xenobiotic substrates from the cytoplasm to the extracellular milieu. Here, we cloned and characterized CsMRP7 as a novel ABC transporter from the Chinese liver fluke, Clonorchis sinensis. Full-length cDNA of CsMRP7 was 5174 nt, encoded 1636 amino acids (aa), and harbored a 147-bp 5'-untranslated region (5'-UTR) and 116-bp 3'-UTR. Phylogenetic analysis confirmed that CsMRP7 was closer to the ABCC subfamily than the ABCB subfamily. Tertiary structures of the N-terminal region (1-322 aa) and core region (323-1621 aa) of CsMRP7 were generated by homology modeling using glucagon receptor (PDB ID: 5ee7_A) and P-glycoprotein (PDB ID: 4f4c_A) as templates, respectively. CsMRP7 nucleotide-binding domain 2 (NBD2) was conserved more than NBD1, which was the sites of ATP binding and hydrolysis. Like typical long MRPs, CsMRP7 has an additional membrane-spanning domain 0 (MSD0) and cytoplasmic loop, along with a common structural fold consisting of MSD1-NBD1-MSD2-NBD2 as a single polypeptide assembly. MSD0, MSD1, and MSD2 consisted of TM1-7, TM8-13, and TM14-19, respectively. The CsMRP7 transcript was more abundant in the metacercariae than in the adult worms. Truncated NBD1 (39 kDa) and NBD2 (44 kDa) were produced in bacteria and mouse immune sera were raised. CsMRP7 was localized in the apical side of the intestinal epithelium, sperm in the testes and seminal receptacle, receptacle membrane, and mesenchymal tissue around intestine in the adult worm. These results provide molecular information and insights into structural and functional characteristics of CsMRP7 and homologs of flukes.

Expression profiles of glyceraldehyde-3-phosphate dehydrogenase from Clonorchis sinensis: a glycolytic enzyme with plasminogen binding capacity.

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Hu, Yue; Zhang, Erhong; Huang, Lisi; Li, Wenfang; Liang, Pei; Wang, Xiaoyun; Xu, Jin; Huang, Yan; Yu, Xinbing

2014-12-01

Globally, 15-20 million people are infected with Clonorchis sinensis (C. sinensis) which results in clonorchiasis. In China, clonorchiasis is considered to be one of the fastest-growing food-borne parasitic diseases. That more key molecules of C. sinensis are characterized will be helpful to understand biology and pathogenesis of the carcinogenic liver fluke. Glyceraldehyde-3-phosphate dehydrogenases (GAPDHs) from many species have functions other than their catalytic role in glycolysis. In the present study, we analyzed the sequence and structure of GAPDH from C. sinensis (CsGAPDH) by using bioinformatics tools and obtained its recombinant protein by prokaryotic expression system, to learn its expression profiles and molecular property. CsGAPDH could bind to human intrahepatic biliary epithelial cell in vivo and in vitro by the method of immunofluorescence assays. CsGAPDH also disturbed in lumen of biliary tract near to the parasite in the liver of infected rat. Western blotting analysis together with immunofluorescence assay indicated that CsGAPDH was a component of excretory/secretory proteins (CsESPs) and a surface-localized protein of C. sinensis. Quantitative real-time PCR (Q-PCR) and Western blotting demonstrated that CsGAPDHs are expressed at the life stages of adult worm, metacercaria, and egg, but the expression levels were different from each other. Recombinant CsGAPDH (rCsGAPDH) was confirmed to have the capacity to catalyze the conversion of glyceraldehyde 3-phosphate to D-glycerate 1,3-bisphosphate which was inhibited by AMP in a dose-dependent manner. In addition, rCsGAPDH was able to interact with human plasminogen in a dose-dependent manner by ELISA. The interaction could be inhibited by lysine. The plasminogen binding capacity of rCsGAPDH along with the distribution of CsGAPDH in vivo and in the liver of C. sinensis-infected rat hinted that surface-localized CsGAPDH might play an important role in host invasion of the worm besides its glycolytic

Identification of microRNA genes in three opisthorchiids.

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Vladimir Y Ovchinnikov

2015-04-01

Full Text Available Opisthorchis felineus, O. viverrini, and Clonorchis sinensis (family Opisthorchiidae are parasitic flatworms that pose a serious threat to humans in some countries and cause opisthorchiasis/clonorchiasis. Chronic disease may lead to a risk of carcinogenesis in the biliary ducts. MicroRNAs (miRNAs are small noncoding RNAs that control gene expression at post-transcriptional level and are implicated in the regulation of various cellular processes during the parasite- host interplay. However, to date, the miRNAs of opisthorchiid flukes, in particular those essential for maintaining their complex biology and parasitic mode of existence, have not been satisfactorily described.Using a SOLiD deep sequencing-bioinformatic approach, we identified 43 novel and 18 conserved miRNAs for O. felineus (miracidia, metacercariae and adult worms, 20 novel and 16 conserved miRNAs for O. viverrini (adult worms, and 33 novel and 18 conserved miRNAs for C. sinensis (adult worms. The analysis of the data revealed differences in the expression level of conserved miRNAs among the three species and among three the developmental stages of O. felineus. Analysis of miRNA genes revealed two gene clusters, one cluster-like region and one intronic miRNA in the genome. The presence and structure of the two gene clusters were validated using a PCR-based approach in the three flukes.This study represents a comprehensive description of miRNAs in three members of the family Opistorchiidae, significantly expands our knowledge of miRNAs in multicellular parasites and provides a basis for understanding the structural and functional evolution of miRNAs in these metazoan parasites. Results of this study also provides novel resources for deeper understanding the complex parasite biology, for further research on the pathogenesis and molecular events of disease induced by the liver flukes. The present data may also facilitate the development of novel approaches for the prevention and

Identification, immunolocalization, and immunological characterization of nitric oxide synthase-interacting protein from Clonorchis sinensis.

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Bian, Meng; Li, Shan; Wang, Xiaoyun; Xu, Yanquan; Chen, Wenjun; Zhou, Chenhui; Chen, Xueqing; He, Lei; Xu, Jin; Liang, Chi; Wu, Zhongdao; Huang, Yan; Li, Xuerong; Yu, Xinbing

2014-05-01

Recently, accumulating evidences indicate that nitric oxide (NO) is a potent mediator with diverse roles in regulating cellular functions, signaling pathways, and variety of pathological processes. In the present study, using data from the published genomic for Clonorchis sinensis (C. sinensis), we investigated a gene encoding nitric oxide synthase-interacting protein (NOSIP) of C. sinensis. Recombinant CsNOSIP (rCsNOSIP) was expressed and purified from Escherichia coli BL21. The open reading frame of CsNOSIP comprises 867 bp which encodes 289 amino acids and shares 72.9, 45.2, 47, 46.4, and 45.8% identity with NOSIP from Schistosoma mansoni, Xenopus laevis, Rattus norvegicus, Mus musculus, and Homo sapiens, respectively. Bioinformatics analysis suggested that the full-length sequence contains an eNOS-interacting domain and numerous B-cell epitopes. Quantitative RT-PCR indicated that CsNOSIP differentially transcribed throughout the adult worms, metacercariae, and egg stages of C. sinensis, and were highly expressed in the adult worms. Moreover, western blot analysis showed that the rCsNOSIP could be detected by the serum from BALB/c mice infected with C. sinensis and the serum from BALB/c mice immunized with excretory/secretory products (ESPs). Furthermore, immunolocalization assay showed that CsNOSIP was specifically localized in the intestine, vitellarium, and eggs of adult worm. Both immunoblot and immunolocalization results demonstrated that CsNOSIP was one component of ESPs of C. sinensis, which could be supported by SignalP analysis. Moreover, analysis of the antibody subclass and cytokine profile demonstrated that subcutaneously immunized BALB/c mice with rCsNOSIP could significantly enhance serum IgG1 level and up-regulate expression of IL-4 and IL-6 in the splenocytes. Our results suggested that CsNOSIP was an important antigen exposed to host immune system and probably involved in immune regulation of host by inducing Th2-polarized immune response.

Characterization of a gut-associated asparaginyl endopeptidase of Clonorchis sinensis.

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Kang, Jung-Mi; Lee, Jinyoung; Ju, Hye-Lim; Ju, Jung Won; Kim, Jong-Hyun; Pak, Jhang Ho; Kim, Tong-Soo; Hong, Yeonchul; Sohn, Woon-Mok; Na, Byoung-Kuk

2015-06-01

Asparaginyl endopeptidases (AEP: EC 3.4.22.34) are a family of cysteine proteases classified into the MEROPS clan CD, family C13. In this study, we characterized the biochemical and antigenic properties of an AEP of Clonorchis sinensis (CsAEP). The recombinant CsAEP showed hydrolytic activity at pH values ranging from acidic to neutral with optimum activity at pH 6.0. While the recombinant CsAEP was stable at neutral pHs, it was unstable at acidic pHs and resulted in loss of enzymatic activity. The recombinant enzyme was effectively inhibited by iodoacetic acid and N-ethylmaleimide, but not by E-64. The partially purified native CsAEP showed biochemical properties similar to the recombinant enzyme. Native CsAEP is likely to be cleaved into an N-terminal mature enzyme and a C-terminal fragment via autocatalytic activation at acidic pHs. Polyclonal antibody raised against the recombinant CsAEP recognized three forms of CsAEP, proenzyme, the N-terminal mature enzyme and the C-terminal fragment, in the worm extract (WE) of C. sinensis. However, only the C-terminal fragment was mainly found in the excretory and secretory (ES) products of the parasite. Strong CsAEP activity was found in the WE, but only a trace level of CsAEP activity was detected in the ES products of the parasite. CsAEP was expressed in various developmental stages of C. sinensis, from metacercariae to adults, and was found to be localized in the intestine of the parasite as well as in intestinal contents. Sera from rats experimentally infected with C. sinensis reacted with CsAEP beginning 4 weeks after infection. These results suggest that CsAEP is a gut-associated enzyme synthesized in the intestine of C. sinensis and subsequently secreted into the intestinal lumen of the parasite. Copyright © 2015 Elsevier Inc. All rights reserved.

No Paragonimus in high-risk groups in Côte d'Ivoire, but considerable prevalence of helminths and intestinal protozoon infections.

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Traoré, Sylvain G; Odermatt, Peter; Bonfoh, Bassirou; Utzinger, Jürg; Aka, N'da D; Adoubryn, Koffi D; Assoumou, Aka; Dreyfuss, Gilles; Koussémon, Marina

2011-06-03

Paragonimiasis is a neglected tropical disease caused by an infection with lung flukes that is transmitted through the consumption of undercooked crabs. The disease is often confused with tuberculosis. Paragonimiasis is thought to be endemic in south-western Côte d'Ivoire. Two cross-sectional surveys were carried out in the first half of 2009 in patients attending two tuberculosis centres of Abidjan. A third cross-sectional survey was conducted in May 2010 in children of two primary schools in Dabou, where crabs are frequently consumed. Patients with chronic cough provided three sputum samples plus one stool sample. Sputum samples were examined for tuberculosis with an auramine staining technique and for Paragonimus eggs using a concentration technique. Stool samples were subjected to the Ritchie technique. Schoolchildren provided a single stool sample, and samples were subjected to the Kato-Katz and an ether-concentration technique. A pre-tested questionnaire was administered to patients and schoolchildren to investigate food consumption habits. Additionally, between June 2009 and August 2010, shellfish were purchased from markets in Abidjan and Dabou and examined for metacercariae. No human case of paragonimiasis was diagnosed. However, trematode infections were seen in 32 of the 272 shellfish examined (11.8%). Questionnaire results revealed that crab and pig meat is well cooked before consumption. Among the 278 patients with complete data records, 62 had tuberculosis, with a higher prevalence in males than females (28.8% vs. 13.9%, χ2 = 8.79, p = 0.003). The prevalence of helminths and intestinal protozoa was 4.6% and 16.9%, respectively. In the school survey, among 166 children with complete data records, the prevalence of helminths and intestinal protozoa was 22.3% and 48.8%, respectively. Boys had significantly higher prevalences of helminths and intestinal protozoa than girls. Hookworm was the predominant helminth species and Entamoeba coli was the most

Molecular cloning and characterization of Fasciola gigantica thioredoxin-glutathione reductase.

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Changklungmoa, Narin; Kueakhai, Pornanan; Sangpairoj, Kant; Chaichanasak, Pannigan; Jaikua, Wipaphorn; Riengrojpitak, Suda; Sobhon, Prasert; Chaithirayanon, Kulathida

2015-06-01

The Fasciola gigantica thioredoxin-glutathione reductase (FgTGR) gene is a fusion between thioredoxin reductase (TR) and a glutaredoxin (Grx) gene. FgTGR was cloned by polymerase chain reaction (PCR) from adult complementary DNA (cDNA), and its sequences showed two isoforms, i.e., the cytosolic and mitochondrial FgTGR. Cytosolic FgTGR (cytFgTGR) was composed of 2370 bp, and its peptide had no signal sequence and hence was not a secreted protein. Mitochondrial FgTGR (mitFgTGR) was composed of 2506 bp with a signal peptide of 43 amino acids; therefore, it was a secreted protein. The putative cytFgTGR and mitFgTGR peptides comprised of 598 and 641 amino acids, respectively, with a molecular weight of 65.8 kDa for cytFgTGR and mitFgTGR, with a conserved sequence (CPYC) of TR, and ACUG and CVNVGC of Grx domains. The recombinant FgTGR (rFgTGR) was expressed in Escherichia coli BL21 (DE3) and used for production for a polyclonal antibody in rabbits (anti-rFgTGR). The FgTGR protein expression, estimated by indirect ELISA using the rabbit anti-rFgTGR as probe, showed high levels of expression in eggs, and 2- and 4-week-old juveniles and adults. The rFgTGR exhibited specific activities in the 5,5'-dithiobis (2-nitro-benzoic acid) (DTNB) reductase assay for TR activity and in β-hydroxyethul disulfide (HED) for Grx activity. When analyzed by immunoblotting and immunohistochemistry, rabbit anti-rFgTGR reacted with natural FgTGR at a molecular weight of 66 kDa from eggs, whole body fraction (WB) of metacercariae, NEJ, 2- and 4-week-old juveniles and adults, and the tegumental antigen (TA) of adult. The FgTGR protein was expressed at high levels in the tegument of 2- and 4-week-old juveniles. The FgTGR may be one of the major factors acting against oxidative stresses that can damage the parasite; hence, it could be considered as a novel vaccine or a drug target.

Protection against Fasciola gigantica infection in mice by vaccination with recombinant juvenile-specific cathepsin L.

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Sansri, Veerawat; Meemon, Krai; Changklungmoa, Narin; Kueakhai, Pornanan; Chantree, Pathanin; Chaichanasak, Pannigan; Lorsuwannarat, Natcha; Itagaki, Tadashi; Sobhon, Prasert

2015-03-24

Fasciola gigantica cathepsin L1H (FgCatL1H) is one of the major cathepsin L released by juveniles of F. gigantica to aid in the invasion of host's tissues. Due to its high sequence similarity with other cathepsin L (CatL) isoforms of late stage F. gigantica, it was considered to be a good vaccine candidate that can block all CatL-mediated protease activities and affect juveniles as well as adult parasites. In this study, recombinant proFgCatL1H protein expressed in yeast, Pichia pastoris, system was mixed with Freund's adjuvants and used to subcutaneously immunize mice that were later challenged with metacercariae of F. gigantica. The percentage of worm protection in the rproFgCatL1H-vaccinated mice compared to the non-immunized and adjuvant control mice were approximately 62.7% and 66.1%, respectively. Anti-rproFgCatL1H antisera collected from vaccinated mice reacted specifically with rproFgCatL1H and other cathepsin L isoforms of F. gigantica, but the antibodies did not cross react with antigens from other trematode and nematode parasites, including Eurytrema pancreaticum, Opisthorchis viverrini, Fischoederius cobboldi, Cotylophoron cotylophorum, Gigantocotyle explanatum, Paramphistomum cervi, and Setaria labiato-papillosa. The levels of IgG1 and IgG2a in mouse sera increased significantly at two weeks after immunization and were highest during the sixth to eighth weeks after immunization. The IgG1 level was higher than IgG2a at all periods of immunization, implicating the dominance of the Th2 response. The levels of IgG1 and IgG2a in the immune sera were shown to be strongly correlated with the numbers of worm recovery, and the correlation coefficient was higher for IgG1. The levels of serum aspartate aminotransferase and alanine transaminase were significantly lower in the sera of rproFgCatL1H-vaccinated mice than in the infected control mice indicating a lower degree of liver damage. This study demonstrated a high potential of FgCatL1H vaccine, and its

No Paragonimus in high-risk groups in Côte d'Ivoire, but considerable prevalence of helminths and intestinal protozoon infections

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Assoumou Aka

2011-06-01

Full Text Available Abstract Background Paragonimiasis is a neglected tropical disease caused by an infection with lung flukes that is transmitted through the consumption of undercooked crabs. The disease is often confused with tuberculosis. Paragonimiasis is thought to be endemic in south-western Côte d'Ivoire. Methods Two cross-sectional surveys were carried out in the first half of 2009 in patients attending two tuberculosis centres of Abidjan. A third cross-sectional survey was conducted in May 2010 in children of two primary schools in Dabou, where crabs are frequently consumed. Patients with chronic cough provided three sputum samples plus one stool sample. Sputum samples were examined for tuberculosis with an auramine staining technique and for Paragonimus eggs using a concentration technique. Stool samples were subjected to the Ritchie technique. Schoolchildren provided a single stool sample, and samples were subjected to the Kato-Katz and an ether-concentration technique. A pre-tested questionnaire was administered to patients and schoolchildren to investigate food consumption habits. Additionally, between June 2009 and August 2010, shellfish were purchased from markets in Abidjan and Dabou and examined for metacercariae. Results No human case of paragonimiasis was diagnosed. However, trematode infections were seen in 32 of the 272 shellfish examined (11.8%. Questionnaire results revealed that crab and pig meat is well cooked before consumption. Among the 278 patients with complete data records, 62 had tuberculosis, with a higher prevalence in males than females (28.8% vs. 13.9%, χ2 = 8.79, p = 0.003. The prevalence of helminths and intestinal protozoa was 4.6% and 16.9%, respectively. In the school survey, among 166 children with complete data records, the prevalence of helminths and intestinal protozoa was 22.3% and 48.8%, respectively. Boys had significantly higher prevalences of helminths and intestinal protozoa than girls. Hookworm was the

Health assessment of the oyster Crassostrea rhizophorae on the southern coast of Bahia, northeastern Brazil Avaliação da saúde da ostra Crassostrea rhizophorae no Litoral Sul da Bahia, nordeste do Brasil

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Rosana Pinho Brandão

Full Text Available This study investigated the health of natural stocks of the oyster Crassostrea rhizophorae on the southern coast of Bahia in northeastern Brazil, during summer and winter 2010, at three localities (sampling points in the estuaries of the Maraú (Camamu Bay and Graciosa rivers. A total of 180 oysters (30/sampling point/season were examined macroscopically for the presence of pathogens and anatomical changes. The specimens were subsequently fixed in Davidson solution, processed for paraffin embedding, sectioned and stained with Harris' hematoxylin and eosin. Histological analysis revealed the presence of Rickettsia-like organisms (RLOs, Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus in the sporocyst phase, a nonspecific metacercaria, and a metacestode of genus Tylocephalum. The prevalence of infection was low except for parasitism by Nematopsis sp. which also caused histopathological changes. The presence of Bucephalus sp. caused parasitic castration. These two pathogens significantly affect the health of C. rhizophorae.Este estudo investigou a saúde de ostras da espécie Crassostrea rhizophorae de estoques naturais do Litoral Sul do Estado da Bahia, Nordeste do Brasil, durante o verão e o inverno de 2010, em três pontos amostrais distribuídos nos estuários dos rios Maraú (Baía de Camamu e Graciosa. Um total de 180 ostras (30/ponto amostral/período foram examinadas macroscopicamente para a presença de patógenos e alterações anatômicas e posteriormente fixadas em solução de Davidson, processadas para inclusão em parafina, seccionadas e coradas com hematoxilina de Harris e eosina. A análise histológica evidenciou a presença de organismos com características similares a Rickettsia (RLOs, Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus em fase esporocística, metacercária inespecífica e metacestóide de Tylocephalum. As prevalências de infecção foram baixas, com exceção do

Clonorchis sinensis acetoacetyl-CoA thiolase: identification and characterization of its potential role in surviving in the bile duct.

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Lin, Jinsi; Qu, Hongling; Chen, Guishan; He, Lei; Xu, Yanquan; Xie, Zhizhi; Ren, Mengyu; Sun, Jiufeng; Li, Shan; Chen, Wenjun; Chen, Xueqing; Wang, Xiaoyun; Li, Xuerong; Liang, Chi; Huang, Yan; Yu, Xinbing

2015-02-25

Clonorchis sinensis (C. sinensis) inhabits in bile duct of the host. However, the mechanisms involved in why C. sinensis can survive in the bile environment containing lipids have not yet been explored. In this study, C. sinensis acetoacetyl-CoA thiolase (CsACAT), a member of the thiolase family which has a key role in the beta oxidation pathway of fatty acid production, was identified and characterized to understand its potential role in adapting to the bile environment. The encoding sequence, conserved domains and spatial structure of CsACAT were identified and analyzed by bioinformatic tools. Recombinant CsACAT (rCsACAT) was obtained using a procaryotic expression system. The expression pattern of CsACAT was confirmed by quantitative real-time PCR, western blotting, and immunofluorescence. Gradients of lecithin were then set to culture C. sinensis adults in vitro and the survival rate of C. sinensis was analyzed, as well as the expression level and enzymatic activity of CsACAT in different lipid environments. Hypercholesteremia rabbit models were established by feeding with a hyperlipidemic diet and then infected intragastrically with C. sinensis. One and a half months later, the worm burdens and the expression level of CsACAT was detected. CsACAT was confirmed to be a member of the thiolase family and present in the excretory/secretory proteins of C. sinensis. CsACAT was specifically localized at the vitellarium and sub-tegumental muscle layer in adult worms. The mRNA level of CsACAT in eggs was higher than those in adult worms and metacercariae. When adult worms were cultured with higher concentration of lecithin, the expression level and enzyme activity of CsACAT were up-regulated. The survival rate of adult worms was higher than control group. More adult worms were recovered from hypercholesteremia rabbit models. The expression level of CsACAT in these worms was higher than control group. Our results implied that C. sinensis might sense lipid levels and

Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target

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Yu-Jung Kim

2017-03-01

Full Text Available The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6 was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis. Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis, CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results

Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target.

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Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won

2017-03-04

The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis . However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6) was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis . Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis , CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results contribute to a

Parasites of the flounder Platichthys flesus (L.) from the German Bight, North Sea, and their potential use in ecosystem monitoring. A. Infection characteristics of potential indicator species

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Schmidt, V.; Zander, S.; Körting, W.; Steinhagen, D.

2003-10-01

As part of integrated biological-effect monitoring, the parasite fauna of the flounder Platichthys flesus (L.) was investigated at five locations in the German Bight, with a view to using parasite species as bio-indicators. Over a period of 6 years, parasites from 30 different taxa were identified, but only 7 taxa of the parasite community occurred regularly at all locations and in sufficient abundance that they could be considered as potential indicator species. These species were the ciliophoran Trichodina spp., the copepods Acanthochondria cornuta, Lepeophtheirus pectoralis and Lernaeocera branchialis, the helminths Zoogonoides viviparus and Cucullanus heterochrous and metacercaria of an unidentified digenean species. Infection characteristics of these parasites are presented, with a comparison of the results from individual sampling periods and those of the long-term data set. Natural influences on the infection levels, such as temporal variations, habitat conditions and host-related factors, were evaluated. All of these parasite species showed significant differences in their infection levels between the Elbe estuary, as the most polluted site, and the less polluted coastal and marine locations: Helgoland, Outer Eider estuary and Spiekeroog, especially in the long-term data set. Gradual differences between the Elbe, the Outer Eider and Helgoland, which were not detected in individual sampling periods, also became evident in the pooled-data set. These were found in the prevalence of Trichodina spp., A. cornuta, Z. viviparus and C. heterochrous. Although salinity is considered as the most important natural factor, influencing the distribution pattern of the majority of the potential indicator species, infection levels of most of these species differed between locations with similar salinity conditions. Infection levels corresponded to a contamination gradient (Elbe > Inner Eider, Outer Eider > Helgoland) established across the locations. Seasonal variation in

Determination Of Irradiation Doses For Fasciola gigantica (Liver Flukes) For Protection In Goat (Capra hircus Linn.)

International Nuclear Information System (INIS)

Tuasikal, B.J.; Arifin, M.; Tarmizi

2002-01-01

An experiment was conducted to study the cange of pathogenicity of 6 O C o-irradiated F. gigantica that give immunological responses in goat. The aim of the study was to see immunity respond of goats against F. gigantica, which at the end to develop a new strategic technique for fluke control in small ruminant. Irradiated and non-irradiated -using y-cell of 6 0 C o -metacercariae (mc.) of F.gigantica were used as the infection material that administered into 18 locally bred goats averagely 10 months of age. These animal are divided into 6 groups of treatment as the following : (I) infection with 0 Gy irradiated mc as positive control; (2) without any infection, as negative contral; (3) infection with 25 Gy inaiiated mc; (4) immunisation with 25 Gy irradiated mc and then challenge by pure infective mc; (5) infection with 35 Gy irradiated mc; and (6) immunisation with 35 Gy irradiated mc an then challenge by pure infective mc. A period of 8 weeks was given to challenge immunisation. Body weight change, blood value (eosinophyl, haemoglobin, PCV, and reticulocyte) and liver anatomical pathology (AP) are observed and recorded. Result of this experiment showed a reduction in body weight change, 130 grams/week, in group I as compare to animal in group 4, which gain body weight of 80 grams/week. The proportion of eosinophyl in group 4 found to re higher as compare to group 6,90.6% and 66.8%, respectively, which indicates that 25Gy mc immunization enables elimination of the mc challenge given. This condition was confirmed by the ''normal'' seen on the anatomical pathology of the liver as compare to animal in- group 1, which found to be more cirrhosis with eosinophyl proportion of 98,6%. Furthermore, results from the observation of PCV and Hb indicate that anaemia was found in animal from group 1 compare to the respective groups: 22.33% and 8.94% vs 25.90% and 10.10% and 27.10% and 10.10%, respectively for PCV and HB in-group animal 1,4 and 6. It can be concluded that mc

A geographic information system forecast model for strategic control of fasciolosis in Ethiopia.

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Yilma, J M; Malone, J B

1998-07-31

A geographic information system (GIS) forecast model based on moisture and thermal regime was developed to assess the risk of Fasciola hepatica, a temperate species, and its tropical counterpart, Fasciola gigantica, in Ethiopia. Agroecological map zones and corresponding environmental features that control the distribution and abundance of the disease and its snail intermediate hosts were imported from the Food and Agriculture Organization (FAO) Crop Production System Zones (CPSZ) database on east Africa and used to construct a GIS using ATLAS GIS 3.0 software. Base temperatures of 10 degrees C and 16 degrees C were used for F. hepatica and F. gigantica, respectively, to calculate growing degree days in a previously developed climate forecast system that was modified to allow use of monthly climate data values. The model was validated by comparison of risk indices and environmental features to available survey data on fasciolosis. Monthly Fasciola risk indices of four climatic regions in Ethiopia were used to project infection transmission patterns under varying climatic conditions and strategic chemotherapeutic fasciolosis control schemes. Varying degrees of F. hepatica risk occurred in most parts of the country and distinct regional F. hepatica transmission patterns could be identified. In the humid west, cercariae-shedding was predicted to occur from May to October. In the south it occurred from April to May and September to October, depending on the annual abundance of rain. In the north-central and central regions, risk was highest during heavy summer rains and pasture contamination with metacercariae was predicted to occur during August-September, except in wet years, when it may start as early as July and extend up to October. At cooler sites above altitude of 2800 m, completion of an infection cycle may require more than a year. Fasciola gigantica risk was present in the western, southern and north-central regions of the country at altitudes of 1440-2560 m

Understanding the main route of drug entry in adult Fasciola hepatica: Further insights into closantel pharmacological activity.

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Ceballos, L; Canton, C; Cadenazzi, G; Larsen, K; Virkel, G; Moreno, L; Fairweather, I; Lanusse, C; Alvarez, L

2017-10-01

Closantel (CLS) is highly effective against adult liver flukes after its oral or subcutaneous (sc) administration in ruminants. Trans-tegumental diffusion and oral ingestion are the two potential routes available for the entry of drugs into Fasciola hepatica. The work reported here contributes to improve the understanding of CLS pharmacology. The main goals of were: I) to determine the pattern of in vivo CLS accumulation into adult F. hepatica and relevant tissues in CLS-treated sheep; II) to investigate the influence of the physicochemical composition of the incubation medium on the CLS diffusion process into adult F. hepatica; III) to assess the ovicidal activity of CLS against F. hepatica eggs; and IV) to investigate the in vivo effect of CLS treatment on glutathione S-transferases activity in adult liver flukes exposed to CLS. Fourteen healthy sheep were each orally infected with 75 F. hepatica metacercariae. Sixteen (16) weeks after infection, animals were treated with CLS by oral (n = 6, 10 mg/kg) or sub-cutaneous (sc) (n = 6, 5 mg/kg) route. At 12, 24 and 36 h post-treatment, animals were sacrificed (n = 2) and samples of blood, bile and adult F. hepatica were collected. In addition, flukes recovered from non-treated sheep (n = 2) were ex vivo incubated (60 min) in the presence of CLS in either RPMI or bile as incubation medium. CLS concentration was measured by HPLC. The ovicidal activity of CLS was investigated using eggs obtained from the bile of untreated sheep. Finally, glutathione S-transferase activity in F. hepatica recovered from untreated and CLS-treated sheep was assessed. In the in vivo studies, the highest CLS concentrations were measured in plasma and adult liver flukes. A positive correlation was observed between CLS concentration in plasma and in F. hepatica. Results obtained in the current work indicate that the in vivo accumulation of CLS into adult liver flukes occurs mainly by the oral route. After ex

Effects of Triclabendazole and Al bendazole on treatment of Fascioliasis

International Nuclear Information System (INIS)

Mawoad, M.A.F.; Amin, M.M.; Elnashar, D. E.

2010-01-01

Fascioliasis is a zoonotic disease caused by the trematode Fasciola hepatica, a liver fluke. It affect numerous species mainly ruminants. Its usual hosts are sheep or cattle, in which it causes a disease of economic importance. The prevalence of human infection has been reported worldwide. The present study was designed to evaluate the comparative efficacy of different anthelmintic drugs against Fasciola hepatica infestation. Twenty parasite-free rabbits were divided into four groups, group I: non-infected non- treated rabbits, kept as negative control. Rabbits of group II, III and IV were intended for infection with 20 Fasciola hepatica metacercariae per animal. Group II received no treatment and kept as positive control. Animals of group III administered Triclabendazole (TCBZ) treatment orally in a single dose (0.45 mg/Kg b. wt.) 60 days after infection. Consistent with group III, animals of group IV administered a bendazole (ABZ) treatment orally in a single dose (0.52 mg/Kg b.wt.) The comparisons between the anthelmintic efficacy of TCBZ and ABZ were done throughout (1): parasitological evaluation of egg counts; (2): biochemical evaluation of serum AST and ALT; (3): histopathological examination of liver and (4): scanning electron microscope of host worms. Data represented showed that the egg count was highly significant reduced in rabbits treated with TCBZ as compared with those of positive control group. But, rabbits treated with ABZ revealed slightly reduction as compared with those of positive control group. Also, the results illustrated that infection of rabbits with Fasciola hepatica induced a highly significant increased in serum (AST and ALT) activities compared to control group. In response to treatment with TCBZ and ABZ, it was revealed a significant decrease in the enzyme activity of both serum AST and ALT levels compared to those of control negative group as well as control positive group. However, histopathological examination showed a significant

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis.

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Yang, Qing-Li; Shen, Ji-Qing; Jiang, Zhi-Hua; Shi, Yun-Liang; Wan, Xiao-Ling; Yang, Yi-Chao

2017-08-07

Although the responses of inducible nitric oxide synthase (iNOS) and associated cytokine after Clonorchis sinensis infection have been studied recently, their mechanisms remain incompletely understood. In this study, we investigated the effects of toll-like receptor 2 (TLR2) signals on iNOS/nitric oxide (NO) responses after C. sinensis infection. We also evaluated the correlations between iNOS responses and worm development, which are possibly regulated by TLR2 signal. TLR2 wild-type and mutant C57BL/6 J mice were infected with 60 C. sinensis metacercariae, and the samples were collected at 30, 60, 90 and 120 days post-infection (dpi). The total serum NO levels were detected using Griess reagent after nitrate was reduced to nitrite. Hepatic tissue samples from the infected mice were sliced and stained with hematoxylin and eosin (HE) to observe worm development in the intrahepatic bile ducts. The iNOS mRNA transcripts in the splenocytes were examined by real time reverse transcriptase polymerase chain reaction (qRT-PCR), and iNOS expression was detected by immunohistochemistry. Developing C. sinensis juvenile worms were more abundant in the intrahepatic bile ducts of TLR2 mutant mice than those of TLR2 wild-type mice. However, no eggs were found in the faeces of both mice samples. The serum levels of total NO significantly increased in TLR2 mutant mice infected with C. sinensis at 30 (t (5)  = 2.595, P = 0.049), 60 (t (5)  = 7.838, P = 0.001) and 90 dpi (t (5)  = 3.032, P = 0.029). Meanwhile, no changes occurred in TLR2 wild-type mice compared with uninfected controls during the experiment. The iNOS expression in splenocytes showed unexpected higher background levels in TLR2 mutant mice than those in TLR2 wild-type mice. Furthermore, the iNOS mRNA transcripts in splenocytes were significantly increased in the TLR2 wild-type mice infected with C. sinensis at 30 (t (5)  = 5.139, P = 0.004), 60 (t (5)  = 6.138, P = 0.002) and 90 dpi (t (5)  = 6

A novel ex vivo immunoproteomic approach characterising Fasciola hepatica tegumental antigens identified using immune antibody from resistant sheep.

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Cameron, Timothy C; Cooke, Ira; Faou, Pierre; Toet, Hayley; Piedrafita, David; Young, Neil; Rathinasamy, Vignesh; Beddoe, Travis; Anderson, Glenn; Dempster, Robert; Spithill, Terry W

2017-08-01

4 IgG is either recognising individual proteins released from extracellular vesicles or is immunoprecipitating intact exosome-like extracellular vesicles. Five extracellular vesicle membrane proteins were identified including two proteins predicted to be associated with vesicle transport/ exocytosis (VPS4, vacuolar protein sorting-associated protein 4b and the Niemann-Pick C1 protein). RNAseq analysis of the developmental transcription of the 38 immunosloughate proteins showed that the sequences are expressed over a wide abundance range with 21/38 transcripts expressed at a relatively high level from metacercariae to the adult life cycle stage. A notable feature of the immunosloughates was the absence of cytosolic proteins which have been reported to be secreted markers for damage to adult flukes incubated in vitro, suggesting that the proteins observed are not inadvertent contaminants leaking from damaged flukes ex vivo. The identification of tegument protein antigens shared between F. gigantica and F. hepatica is beneficial in terms of the possible development of a dual purpose vaccine effective against both fluke species. Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

Centrocestus formosanus (Opisthorchiida: Heterophyidae como causa de muerte de alevines de tilapia gris Oreochromis niloticus (Perciforme: Cichlidae en el Pacífico seco de Costa Rica

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Donald Arguedas Cortés

2010-12-01

Full Text Available Centrocestus formosanus es un parásito trematodo zoonótico originario de Asia asociado con muertes de peces principalmente de cultivo. 907 moluscos provenientes de estanques sembrados con tilapias, seleccionados uno por provincia fueron identificados al nivel taxonómico especifico. Se identificaron cuatro gastrópodos y un bivalvo: M. tuberculata, M. turricula, P. flagellata, H. cubensis y A. luteola. Se reporta, por primera vez, la presencia de dos especies de moluscos en Costa Rica. Se identificaron siete morfotipos de cercarias parasitando las cinco especies de moluscos encontradas. En la segunda exposición experimental se demostró que el morfotipo parapleurolofocercus encontrado en M. tuberculata concuerda con el hallazgo de C. formosanus en alevines de tilapia, después del examen clínico, anatomopatológico y parasitológico realizado a los alevines expuestos. Las metacercarias fueron extraídas del quiste utilizando microagujas y micropinzas lavadas en solución salina fisiológica (0.65%, fijadas en formol caliente al 4% y después esquematizadas con una cámara clara adaptada a un microscopio fotónico, estimándose una abundancia e intensidad media de 1018-1027 digeneos por branquia en cada pez parasitado, determinándose así el hospedador intermediario primario y secundario del parásito. En el presente trabajo se reporta por primera vez Centrocestus formosanus en Costa Rica.Centrocestus formosanus (Opisthorchiida: Heterophyidae as a cause of death in gray tilapia fry Oreochromis niloticus (Perciforme: Cichlidae in the dry Pacific of Costa Rica. Centrocestus formosanus is a zoonotic trematode from Asia and has been mainly associated as cause of death of cultured fish. To identify pathogen trematode species in tilapia fry (Oreochromis niloticus and to determine mollusks hosting these parasites, freshwater mollusks were collected from tilapia cultured ponds and experimental infections were carried out with tilapia fries and