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Sample records for fasciola trematoda fasciolidae

  1. Development and evaluation of a single-step duplex PCR for simultaneous detection of Fasciola hepatica and Fasciola gigantica (family Fasciolidae, class Trematoda, phylum Platyhelminthes).

    PubMed

    Le, Thanh Hoa; Nguyen, Khue Thi; Nguyen, Nga Thi Bich; Doan, Huong Thi Thanh; Le, Xuyen Thi Kim; Hoang, Chau Thi Minh; De, Nguyen Van

    2012-08-01

    A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of Fasciola hepatica and F. gigantica. These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of cox1-trnT-rrnL, two species-specific forward primers were designed, FHF (for F. hepatica) and FGF (for F. gigantica), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of Fasciola DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for F. hepatica and 615 bp for F. gigantica. The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each Fasciola species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32 Fasciola samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of Fasciola species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of Fasciola species in areas of distributional and zonal overlap.

  2. Experimental Life History and Biological Characteristics of Fasciola gigantica (Digenea: Fasciolidae)

    PubMed Central

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

    2015-01-01

    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. PMID:25748710

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

    PubMed

    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.

  4. A novel monoclonal antibody-based immunoenzymatic assay for epidemiological surveillance of the vector snails of Fasciola hepatica (Trematoda: Digenea).

    PubMed

    Alba, Annia; Hernández, Hilda M; Marcet, Ricardo; Vázquez, Antonio A; Figueredo, Mabel; Sánchez, Jorge; Otero, Oscar; Sarracent, Jorge

    2015-02-01

    Fasciolosis is a globally distributed snail-borne disease which requires economic consideration due to its enormous impact on veterinary medicine. During recent decades, this parasitosis has also shown increasing prevalence in human populations worldwide. The dissemination and successful transmission of fasciolosis ultimately depends on the existence of susceptible snails that act as intermediate hosts. Therefore, to accomplish effective control of this disease, surveillance and detection of the infected intermediate host would be essential. The screening of trematodes within snails using classical parasitological examination of the larvae can be unreliable (sensitivity and specificity vary depending on the time of infection and the experience of the observer) and relatively costly when using molecular biological methods during large-scale monitoring. Here we propose a novel monoclonal antibody-based immunoenzymatic assay to detect ongoing Fasciola hepatica infection in lymnaeid snails. Anti-F. hepatica rediae mouse monoclonal antibodies were generated and used to develop a double monoclonal antibody-based ELISA for parasite detection. Fasciola hepatica-infected and uninfected laboratory-reared Galba cubensis and Pseudosuccinea columella were used for assessment of the developed ELISA. Experimentally infected snails were dissected and examined for parasite larvae as the "gold standard" method. Sensitivity results were 100% for both snail species, while specificity was 98% for G. cubensis and 100% for P. columella. No cross-reactivity was detected in lymnaeids infected with Trichobilharzia sp. or Cotylophoron sp. The ELISA enabled detection of the infection from day 8 p.i. in G. cubensis while in P. columella it was noted as early as day 4. To our knowledge no previous immunoassays have been reported to detect helminth-infected snails and the developed sandwich ELISA method is therefore suggested for infection status validation in natural populations of lymnaeid

  5. SPATIAL DISTRIBUTION OF LYMNAEIDAE (MOLLUSCA, BASOMMATOPHORA), INTERMEDIATE HOST OF Fasciola hepatica LINNAEUS, 1758 (TREMATODA, DIGENEA) IN BRAZIL

    PubMed Central

    Medeiros, Camilla; Scholte, Ronaldo Guilherme Carvalho; D'ávila, Sthefane; Caldeira, Roberta Lima; Carvalho, Omar dos Santos

    2014-01-01

    Snails of the family Lymnaeidae act as intermediate hosts in the biological cycle of Fasciola hepatica, which is a biological agent of fasciolosis, a parasitic disease of medical importance for humans and animals. The present work aimed to update and map the spatial distribution of the intermediate host snails of F. hepatica in Brazil. Data on the distribution of lymnaeids species were compiled from the Collection of Medical Malacology (Fiocruz-CMM, CPqRR), Collection of Malacology (MZUSP), “SpeciesLink” (CRIA) network and through systematic surveys in the literature. Our maps of the distribution of lymnaeids show that Pseudosuccinea columella is the most common species and it is widespread in the South and Southeast with few records in the Midwest, North and Northeast regions. The distribution of the Galba viatrix, G. cubensis and G. truncatula showed a few records in the South and Southeast regions, they were not reported for the Midwest, North and Northeast. In addition, in the South region there are a few records for G. viatrix and one occurrence of Lymnaea rupestris. Our findings resulted in the first map of the spatial distribution of Lymnaeidae species in Brazil which might be useful to better understand the fasciolosis distribution and delineate priority areas for control interventions. PMID:24879003

  6. Complete mitochondrial genomes of the ‘intermediate form’ of Fasciola and Fasciola gigantica, and their comparison with F. hepatica

    PubMed Central

    2014-01-01

    Background Fascioliasis is an important and neglected disease of humans and other mammals, caused by trematodes of the genus Fasciola. Fasciola hepatica and F. gigantica are valid species that infect humans and animals, but the specific status of Fasciola sp. (‘intermediate form’) is unclear. Methods Single specimens inferred to represent Fasciola sp. (‘intermediate form’; Heilongjiang) and F. gigantica (Guangxi) from China were genetically identified and characterized using PCR-based sequencing of the first and second internal transcribed spacer regions of nuclear ribosomal DNA. The complete mitochondrial (mt) genomes of these representative specimens were then sequenced. The relationships of these specimens with selected members of the Trematoda were assessed by phylogenetic analysis of concatenated amino acid sequence datasets by Bayesian inference (BI). Results The complete mt genomes of representatives of Fasciola sp. and F. gigantica were 14,453 bp and 14,478 bp in size, respectively. Both mt genomes contain 12 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes, but lack an atp8 gene. All protein-coding genes are transcribed in the same direction, and the gene order in both mt genomes is the same as that published for F. hepatica. Phylogenetic analysis of the concatenated amino acid sequence data for all 12 protein-coding genes showed that the specimen of Fasciola sp. was more closely related to F. gigantica than to F. hepatica. Conclusions The mt genomes characterized here provide a rich source of markers, which can be used in combination with nuclear markers and imaging techniques, for future comparative studies of the biology of Fasciola sp. from China and other countries. PMID:24685294

  7. Fasciola hepatica in capybara.

    PubMed

    Santarém, Vamilton Alvares; Tostes, Raimundo Alberto; Alberti, Haroldo; Sanches, Osimar de Carvalho

    2006-07-01

    The purpose of this report was to register the presence of Fasciola hepatica in an adult female capybara (Hydrocherus hydrocheris). The macro and microscopical characteristics of hepatic lesions, together with the finding of F. hepatica in mature forms indicate the possible infection of capybara by the trematode.

  8. Molecular characterization of Fasciola spp. from the endemic area of northern Iran based on nuclear ribosomal DNA sequences.

    PubMed

    Amor, Nabil; Halajian, Ali; Farjallah, Sarra; Merella, Paolo; Said, Khaled; Ben Slimane, Badreddine

    2011-07-01

    Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered as the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the endemic regions of the North of Iran, Fasciola hepatica and Fasciola gigantica have been previously characterized on the basis of morphometric differences, but the use of molecular markers is necessary to distinguish exactly between species and intermediate forms. Samples from buffaloes and goats from different localities of northern Iran were identified morphologically and then genetically characterized by sequences of the first (ITS-1) and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the ITS of the northern Iranian samples with sequences of Fasciola spp. from GenBank showed that the examined specimens had sequences identical to those of the most frequent haplotypes of F. hepatica (n=25, 48.1%) and F. gigantica (n=20, 38.45%), which differed from each other in different variable nucleotide positions of ITS region sequences, and their intermediate forms (n=7, 13.45%), which had nucleotides overlapped between the two Fasciola species in all the positions. The ITS sequences from populations of Fasciola isolates in buffaloes and goats had experienced introgression/hybridization as previously reported in isolates from other ruminants and humans. Based on ITS-1 and ITS-2 sequences, flukes are scattered in pure F. hepatica, F. gigantica and intermediate Fasciola clades, revealing that multiple genotypes of Fasciola are able to infect goats and buffaloes in North of Iran. Furthermore, the phylogenetic trees based upon the ITS-1 and ITS-2 sequences showed a close relationship of the Iranian samples with isolates of F. hepatica and F. gigantica from different localities of Africa and Asia. In the present study, the intergenic transcribed spacers ITS-1 and ITS-2 showed to be reliable approaches for the genetic

  9. Molecular characterization of Fasciola gigantica from Mauritania based on mitochondrial and nuclear ribosomal DNA sequences.

    PubMed

    Amor, Nabil; Farjallah, Sarra; Salem, Mohamed; Lamine, Dia Mamadou; Merella, Paolo; Said, Khaled; Ben Slimane, Badreddine

    2011-10-01

    Fasciolosis caused by Fasciola hepatica and Fasciola gigantica (Platyhelminthes: Trematoda: Digenea) is considered the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. From Africa, F. gigantica has been previously characterized from Burkina Faso, Senegal, Kenya, Zambia and Mali, while F. hepatica has been reported from Morocco and Tunisia, and both species have been observed from Ethiopia and Egypt on the basis of morphometric differences, while the use of molecular markers is necessary to distinguish exactly between species. Samples identified morphologically as F. gigantica (n=60) from sheep and cattle from different geographical localities of Mauritania were genetically characterized by sequences of the first (ITS-1), the 5.8S, and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA) genes and the mitochondrial Cytochrome c Oxidase I (COI) gene. Comparison of the sequences of the Mauritanian samples with sequences of Fasciola spp. from GenBank confirmed that all samples belong to the species F. gigantica. The nucleotide sequencing of ITS rDNA of F. gigantica showed no nucleotide variation in the ITS-1, 5.8S, and ITS-2 rDNA sequences among all samples examined and those from Burkina Faso, Kenya, Egypt and Iran. The phylogenetic trees based on the ITS-1 and ITS-2 sequences showed a close relationship of the Mauritanian samples with isolates of F. gigantica from different localities of Africa and Asia. The COI genotypes of the Mauritanian specimens of F. gigantica had a high level of diversity, and they belonged to the F. gigantica phylogenically distinguishable clade. The present study is the first molecular characterization of F. gigantica in sheep and cattle from Mauritania, allowing a reliable approach for the genetic differentiation of Fasciola spp. and providing basis for further studies on liver flukes in the African countries.

  10. Ecology of free-living metacercariae (Trematoda).

    PubMed

    Morley, Neil J

    2015-06-01

    The presence of trematodes with a free-living metacercarial stage is a common feature of most habitats and includes important species such as Fasciola hepatica, Parorchis acanthus and Zygocotyle lunata. These trematodes encyst on the surface of an animal or plant that can act as a transport host, which form the diet of the target definitive host. Although these species are often considered individually, they display common characteristics in their free-living biology indicating a shared transmission strategy, yet in comparison to species with penetrative cercariae this aspect of their life cycles remains much overlooked. This review integrates the diverse data and presents a novel synthesis of free-living metacercariae using epibiosis as the basis of a new framework to describe the relationship between transport hosts and parasites. All aspects of their biology during the period that they are metabolically independent of a host are considered, from cercarial emergence to metacercarial excystment.

  11. 2D-PAGE analysis of the soluble proteins of the tropical liver fluke, Fasciola gigantica and biliary amphistome, Gigantocotyle explanatum, concurrently infecting Bubalus bubalis.

    PubMed

    Khan, Yasir A; Khan, Mohd Abdul Hannan; Abidi, S M A

    2016-09-01

    The digenetic trematodes, Fasciola gigantica and Gigantocotyle explanatum, belonging to the family Fasciolidae and Paramphistomidae respectively, have been often found to concurrently infect the liver of Indian water buffalo Bubalus bubalis, causing serious pathological damage to the vital organ, incurring huge economic losses. In the present study the soluble gene products of both F. gigantica and G. explanatum were analyzed by 2 dimensional polyacrylamide gel electrophoresis. The soluble proteomic profile revealed considerable similarity as well as differences in the size, distribution pattern, total number, the isoelectric point (pI) and molecular weight (Mr) of the resolved polypeptide spots. The maximum number of polypeptide spots with a molecular weight range of >10 to 160 kDa were recorded with a pI range of 7-9 followed by pI range of 5-7, 9-10 and 3-5 in both the parasites. However, considerable variation was recorded in the Mr of the polypeptides belonging to each pI range. The genetic heterogeneity could be an obvious contributing factor for such differences but some polypeptides appeared to be conserved in the two species. The molecular similarities and the habitat preference by these worms may be a consequence of microenvironmental cues that guide these flukes to reach their habitat through different routes and establish a successful host-parasite relationship.

  12. 2D-PAGE analysis of the soluble proteins of the tropical liver fluke, Fasciola gigantica and biliary amphistome, Gigantocotyle explanatum, concurrently infecting Bubalus bubalis.

    PubMed

    Khan, Yasir A; Khan, Mohd Abdul Hannan; Abidi, S M A

    2016-09-01

    The digenetic trematodes, Fasciola gigantica and Gigantocotyle explanatum, belonging to the family Fasciolidae and Paramphistomidae respectively, have been often found to concurrently infect the liver of Indian water buffalo Bubalus bubalis, causing serious pathological damage to the vital organ, incurring huge economic losses. In the present study the soluble gene products of both F. gigantica and G. explanatum were analyzed by 2 dimensional polyacrylamide gel electrophoresis. The soluble proteomic profile revealed considerable similarity as well as differences in the size, distribution pattern, total number, the isoelectric point (pI) and molecular weight (Mr) of the resolved polypeptide spots. The maximum number of polypeptide spots with a molecular weight range of >10 to 160 kDa were recorded with a pI range of 7-9 followed by pI range of 5-7, 9-10 and 3-5 in both the parasites. However, considerable variation was recorded in the Mr of the polypeptides belonging to each pI range. The genetic heterogeneity could be an obvious contributing factor for such differences but some polypeptides appeared to be conserved in the two species. The molecular similarities and the habitat preference by these worms may be a consequence of microenvironmental cues that guide these flukes to reach their habitat through different routes and establish a successful host-parasite relationship. PMID:27605808

  13. The revised microRNA complement of Fasciola hepatica reveals a plethora of overlooked microRNAs and evidence for enrichment of immuno-regulatory microRNAs in extracellular vesicles.

    PubMed

    Fromm, B; Trelis, M; Hackenberg, M; Cantalapiedra, F; Bernal, D; Marcilla, A

    2015-09-01

    MicroRNAs (miRNAs) are gene regulators that have recently been shown to down-regulate the immune response via extracellular vesicles in the mammalian host of helminthic parasites. Using the miRNA prediction pipeline miRCandRef, we expanded the current miRNA set of the liver fluke Fasciola hepatica (Platyhelminthes, Trematoda) from 16 to 54 miRNAs (42 conserved and 13 novel). Comparing the cellular expression levels with extracellular vesicles, we found all miRNAs expressed and enriched for miRNAs with immuno-regulatory function, tissue growth and cancer. Our findings support the hypothesis that miRNAs are the molecular mediators of the previously demonstrated immune modulatory function of extracellular vesicles.

  14. Hybridization experiments indicate incomplete reproductive isolating mechanism between Fasciola hepatica and Fasciola gigantica.

    PubMed

    Itagaki, T; Ichinomiya, M; Fukuda, K; Fusyuku, S; Carmona, C

    2011-09-01

    Experiments on hybridization between Fasciola hepatica and Fasciola gigantica were carried out to clarify whether a reproductive isolating mechanism appears between the two Fasciola species. Molecular evidence for hybridization was based on the DNA sequence of the internal transcribed spacer 1 (ITS1) region in nuclear ribosomal DNA, which differs between the species. The results suggested that there were not pre-mating but post-mating isolating mechanisms between the two species. However, viable adults of the hybrids F1 and F2 were produced from both parental F. hepatica and F. gigantica. The hybrids inherited phenotypic characteristics such as ratio of body length and width and infectivity to rats from parental Fasciola hepatica and F. gigantica. These findings suggest that reproductive isolation is incomplete between Fasciola hepatica and F. gigantica. Adults of the hybrids F1 and F2 were completely different in mode of reproduction from aspermic Fasciola forms that occur in Asia and seem to be offspring originated from hybridization between F. hepatica and F. gigantica and to reproduce parthenogenetically. PMID:21767436

  15. Localization of thioredoxin in Fasciola hepatica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thioredoxin is a member of a group of small redox proteins that may protect parasites from the host immune response. Experiments were designed to localize thioredoxin anatomically within the adult and immature stages of Fasciola hepatica. SDS-PAGE and Western blotting were performed to determine t...

  16. Hypothetical granulin-like molecule from Fasciola hepatica identified by bioinformatics analysis.

    PubMed

    Machicado, Claudia; Marcos, Luis A; Zimic, Mirko

    2016-01-01

    Fasciola hepatica is considered an emergent human pathogen, causing liver fibrosis or cirrhosis, conditions that are known to be direct causes of cancer. Some parasites have been categorized by WHO as carcinogenic agents such as Opisthorchis viverrini, a relative of F. hepatica. Although these two parasites are from the same class (Trematoda), the role of F. hepatica in carcinogenesis is unclear. We hypothesized that F. hepatica might share some features with O. viverrini and to be responsible to induce proliferation of host cells. We analyzed the recently released genome of F. hepatica looking for a gene coding a granulin-like growth factor, a protein secreted by O. viverrini (Ov-GRN-1), which is a potent stimulator of proliferation of host cells. Using computational biology tools, we identified a granulin-like molecule in F. hepatica, here termed FhGLM, which has high sequence identity level to Ov-GRN-1 and human progranulin. We found evidence of an upstream promoter compatible with the expression of FhGLM. The FhGLM architecture showed to have five granulin domains, one of them, the domain 3, was homologue to Ov-GRN-1 and human GRNC. The structure of the FhGLM granulin domain 3 resulted to have the overall folding of its homologue the human GRNC. Our findings show the presence of a homologue of a potent modulator of cell growth in F. hepatica that might have, as other granulins, a proliferative action on host cells during fascioliasis. Future experimental assays to demonstrate the presence of FhGLM in F. hepatica are needed to confirm our hypothesis. PMID:27386259

  17. Hypothetical granulin-like molecule from Fasciola hepatica identified by bioinformatics analysis.

    PubMed

    Machicado, Claudia; Marcos, Luis A; Zimic, Mirko

    2016-01-01

    Fasciola hepatica is considered an emergent human pathogen, causing liver fibrosis or cirrhosis, conditions that are known to be direct causes of cancer. Some parasites have been categorized by WHO as carcinogenic agents such as Opisthorchis viverrini, a relative of F. hepatica. Although these two parasites are from the same class (Trematoda), the role of F. hepatica in carcinogenesis is unclear. We hypothesized that F. hepatica might share some features with O. viverrini and to be responsible to induce proliferation of host cells. We analyzed the recently released genome of F. hepatica looking for a gene coding a granulin-like growth factor, a protein secreted by O. viverrini (Ov-GRN-1), which is a potent stimulator of proliferation of host cells. Using computational biology tools, we identified a granulin-like molecule in F. hepatica, here termed FhGLM, which has high sequence identity level to Ov-GRN-1 and human progranulin. We found evidence of an upstream promoter compatible with the expression of FhGLM. The FhGLM architecture showed to have five granulin domains, one of them, the domain 3, was homologue to Ov-GRN-1 and human GRNC. The structure of the FhGLM granulin domain 3 resulted to have the overall folding of its homologue the human GRNC. Our findings show the presence of a homologue of a potent modulator of cell growth in F. hepatica that might have, as other granulins, a proliferative action on host cells during fascioliasis. Future experimental assays to demonstrate the presence of FhGLM in F. hepatica are needed to confirm our hypothesis.

  18. Molecular phylogenetic analysis of Fasciola flukes from eastern India.

    PubMed

    Hayashi, Kei; Ichikawa-Seki, Madoka; Mohanta, Uday Kumar; Singh, T Shantikumar; Shoriki, Takuya; Sugiyama, Hiromu; Itagaki, Tadashi

    2015-10-01

    Fasciola flukes from eastern India were characterized on the basis of spermatogenesis status and nuclear ITS1. Both Fasciola gigantica and aspermic Fasciola flukes were detected in Imphal, Kohima, and Gantoku districts. The sequences of mitochondrial nad1 were analyzed to infer their phylogenetical relationship with neighboring countries. The haplotypes of aspermic Fasciola flukes were identical or showed a single nucleotide substitution compared to those from populations in the neighboring countries, corroborating the previous reports that categorized them in the same lineage. However, the prevalence of aspermic Fasciola flukes in eastern India was lower than those in the neighboring countries, suggesting that they have not dispersed throughout eastern India. In contrast, F. gigantica was predominant and well diversified, and the species was thought to be distributed in the area for a longer time than the aspermic Fasciola flukes. Fasciola gigantica populations from eastern India were categorized into two distinct haplogroups A and B. The level of their genetic diversity suggests that populations belonging to haplogroup A have dispersed from the west side of the Indian subcontinent to eastern India with the artificial movement of domestic cattle, Bos indicus, whereas populations belonging to haplogroup B might have spread from Myanmar to eastern India with domestic buffaloes, Bubalus bubalis.

  19. Natural infections of Clinostomum complanatum (Trematoda: Clinostomatidae) in wild herons and egrets, Tottori Prefecture, Japan.

    PubMed

    Aohagi, Y; Shibahara, T; Machida, N; Yamaga, Y; Kagota, K; Hayashi, T

    1992-07-01

    Nycticorax nycticorax, Ardea cinerea, Egretta garzetta, and Egretta intermedia were naturally infected with Clinostomum complanatum (Trematoda: Clinostomatidae) among fourteen wild herons, seven wild egrets and one wild bittern evaluated at the Veterinary Hospital of Tottori University, Tottori, Japan. The latter three species of heron and egrets are reported for the first time as definitive hosts of this parasite in Japan.

  20. Molecular characterization of Fasciola hepatica from Sardinia based on sequence analysis of genomic and mitochondrial gene markers.

    PubMed

    Farjallah, Sarra; Ben Slimane, Badreddine; Piras, Cristina Maria; Amor, Nabil; Garippa, Giovanni; Merella, Paolo

    2013-11-01

    The aim of the present study is to investigate for the first time the genetic diversity of samples identified morphologically as Fasciola hepatica (Platyhelminthes: Trematoda: Digenea) (n=66) from sheep and cattle from two localities of Sardinia and to compare them with available data from other localities by partial sequences of the first (ITS-1), the 5.8S, and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA) genes, the mitochondrial cytochrome c oxidase subunit I (COI), and nicotinamide adenine dinucleotide dehydrogenase subunit I (ND1) genes. Comparison of the sequences from Sardinia with sequences of Fasciola spp. from GenBank confirmed that all samples belong to the species F. hepatica. The nucleotide sequencing of ITS rDNA showed no nucleotide variation in the ITS-1, 5.8S and ITS-2 rDNA sequences among all Sardinian samples, comparing with two ITS-2 haplotypes in standard F. hepatica, showing a substitution C/T in 20 position 859, reported previously from Tunisia, Algeria, Australia, Uruguay and Spain. The present study shows that in Sardinian sheep and cattle there is the most frequent haplotype (FhITS-H1) of F. hepatica species from South Europe. Considering NDI sequences, the phylogenetic trees showed reliable grouping among the haplotypes of F. hepatica from Sardinia and the mitochondrial lineage I, including the main N1 haplotype, observed previously from Europe (Russia, Belarus, Ukraine and Bulgaria), Armenia, West Africa (Nigeria), America (Uruguay and USA), Asia (Turkey, Japan, and China), Georgia, Turkmenistan, Azerbaijan and Australia. Furthermore, common haplotypes FhCOI-H1 and FhCOI-H2 of F. hepatica from Sardinia also corresponded mostly to the first lineage including the main C1 haplotype reported previously from Eastern European and Western Asian populations, they belonged just to a phylogenically distinguishable clade, as F. hepatica from Australia, France, Turkey, Uruguay, Russia, Armenia, Ukraine, Belarus

  1. [New metacercariae of the genus Tylodelphys (Trematoda, Diplostomatidae) in lacustrine populations of Galaxias maculatus (Teleostei, Galaxiidae)].

    PubMed

    Quaggiotto, E A; Valverde, F

    1992-01-01

    Metacercariae of three new species of Tylodelphys (Trematoda, Diplostomatidae) were found in the brain of Galaxias maculatus (Teleostei, Galaxiidae), at the Meliquina Lake, Neuquen Province, Argentina. These metacercariae are morphologically described as Tylodelphys argentinus n. sp., Tylodelphys barilochensis n. sp. and Tylodelphys crubensis n. sp. This is the first time that a stage of genus Tylodelphys is mentioned parasitizing Galaxias maculatus. Metacercariae of T. barilochen sis n. sp. were also found in G. maculatus brain at the Perito Moreno Lake, Rio Negro Province, Argentina.

  2. Bothrigaster variolaris (Trematoda: Cyclocoelidae) infection in two Florida snail kites (Rostrhamus sociabilis plumbeus)

    USGS Publications Warehouse

    Cole, R.A.; Thomas, N.J.; Roderick, C.L.

    1995-01-01

    Two Florida snail kites (Rostrhamus sociabilis plumbeus, Say) collected in August and December 1993 from Osceola County (28 degrees 15'N, 81 degrees 17'W) and Glades County (26 degrees 45'N, 81 degrees 10'W), Florida (USA), respectively, were infected with Bothrigaster variolaris (Trematoda: Cyclocoelidae); this parasite has not been reported previously from birds in the United States. One bird, a fledgling female, harbored 270 specimens of B. variolaris; the helminths caused airsacculitis and bronchitis, possible contributing factors in the death of this bird. The other bird, a subadult female, was infected with 40 trematodes and had gross and microscopic lesions consistent with trauma as the cause of death.

  3. Fasciola hepatica Infection in an Indigenous Community of the Peruvian Jungle.

    PubMed

    Cabada, Miguel M; Castellanos-Gonzalez, Alejandro; Lopez, Martha; Caravedo, María Alejandra; Arque, Eulogia; White, Arthur Clinton

    2016-06-01

    Fasciola hepatica is a zoonotic infection with a worldwide distribution. Autochthonous cases have not been reported in the Amazon region of Peru. Operculated eggs resembling F. hepatica were identified in the stools of five out of 215 subjects in a remote indigenous community of the Peruvian jungle. Polymerase chain reaction targeting Fasciola hepatica cytochrome oxidase subunit 1 (COI) gene and sequencing of the products confirmed Fasciola infection.

  4. In vitro manipulation of gene expression in larval Schistosoma: a model for postgenomic approaches in Trematoda.

    PubMed

    Yoshino, Timothy P; Dinguirard, Nathalie; Mourão, Marina de Moraes

    2010-03-01

    With rapid developments in DNA and protein sequencing technologies, combined with powerful bioinformatics tools, a continued acceleration of gene identification in parasitic helminths is predicted, potentially leading to discovery of new drug and vaccine targets, enhanced diagnostics and insights into the complex biology underlying host-parasite interactions. For the schistosome blood flukes, with the recent completion of genome sequencing and comprehensive transcriptomic datasets, there has accumulated massive amounts of gene sequence data, for which, in the vast majority of cases, little is known about actual functions within the intact organism. In this review we attempt to bring together traditional in vitro cultivation approaches and recent emergent technologies of molecular genomics, transcriptomics and genetic manipulation to illustrate the considerable progress made in our understanding of trematode gene expression and function during development of the intramolluscan larval stages. Using several prominent trematode families (Schistosomatidae, Fasciolidae, Echinostomatidae), we have focused on the current status of in vitro larval isolation/cultivation as a source of valuable raw material supporting gene discovery efforts in model digeneans that include whole genome sequencing, transcript and protein expression profiling during larval development, and progress made in the in vitro manipulation of genes and their expression in larval trematodes using transgenic and RNA interference (RNAi) approaches.

  5. Genetic characterization, species differentiation and detection of Fasciola spp. by molecular approaches

    PubMed Central

    2011-01-01

    Liver flukes belonging to the genus Fasciola are among the causes of foodborne diseases of parasitic etiology. These parasites cause significant public health problems and substantial economic losses to the livestock industry. Therefore, it is important to definitively characterize the Fasciola species. Current phenotypic techniques fail to reflect the full extent of the diversity of Fasciola spp. In this respect, the use of molecular techniques to identify and differentiate Fasciola spp. offer considerable advantages. The advent of a variety of molecular genetic techniques also provides a powerful method to elucidate many aspects of Fasciola biology, epidemiology, and genetics. However, the discriminatory power of these molecular methods varies, as does the speed and ease of performance and cost. There is a need for the development of new methods to identify the mechanisms underpinning the origin and maintenance of genetic variation within and among Fasciola populations. The increasing application of the current and new methods will yield a much improved understanding of Fasciola epidemiology and evolution as well as more effective means of parasite control. Herein, we provide an overview of the molecular techniques that are being used for the genetic characterization, detection and genotyping of Fasciola spp.. PMID:21658284

  6. Molecular characterization and phylogenetic analysis of Fasciola hepatica from Peru.

    PubMed

    Ichikawa-Seki, Madoka; Ortiz, Pedro; Cabrera, Maria; Hobán, Cristian; Itagaki, Tadashi

    2016-06-01

    The causative agent of fasciolosis in South America is thought to be Fasciola hepatica. In this study, Fasciola flukes from Peru were analyzed to investigate their genetic structure and phylogenetic relationships with those from other countries. Fasciola flukes were collected from the three definitive host species: cattle, sheep, and pigs. They were identified as F. hepatica because mature sperms were observed in their seminal vesicles, and also they displayed Fh type, which has an identical fragment pattern to F. hepatica in the nuclear internal transcribed spacer 1. Eight haplotypes were obtained from the mitochondrial NADH dehydrogenase subunit 1 (nad1) sequences of Peruvian F. hepatica; however, no special difference in genetic structure was observed between the three host species. Its extremely low genetic diversity suggests that the Peruvian population was introduced from other regions. Nad1 haplotypes identical to those of Peruvian F. hepatica were detected in China, Uruguay, Italy, Iran, and Australia. Our results indicate that F. hepatica rapidly expanded its range due to human migration. Future studies are required to elucidate dispersal route of F. hepatica from Europe, its probable origin, to other areas, including Peru.

  7. Molecular evidence shows that the liver fluke Fasciola gigantica is the predominant Fasciola species in ruminants from Pakistan.

    PubMed

    Chaudhry, U; van Paridon, B; Shabbir, M Z; Shafee, M; Ashraf, K; Yaqub, T; Gilleard, J

    2016-03-01

    Fascioliasis is an important disease affecting livestock, with great costs to producers worldwide. It has also become a serious issue for human populations in some endemic areas as an emerging zoonotic infection. There are two Fasciola species of liver fluke responsible for this disease, which occur worldwide, Fasciola hepatica and Fasciola gigantica. Identifying these two species on the basis of adult or egg morphology requires specialist knowledge due to the similarity of characters, and may misidentify putative intermediate or hybrid forms. In this study we sequenced the internal transcribed spacer 2 (ITS-2) rDNA of liver flukes collected from multiple species of hosts from seven localities in the Punjab and Baluchistan provinces of Pakistan, to determine the distribution of these two species. All 46 flukes processed in this study, collected from seven sites, showed the rDNA ITS-2 genotype corresponding to F. gigantica, contradicting previous reports, based on adult and egg morphology, that both species are present in Pakistan, with F. hepatica being the more common.

  8. A case of biliary Fascioliasis by Fasciola gigantica in Turkey.

    PubMed

    Goral, Vedat; Senturk, Senem; Mete, Omer; Cicek, Mutallib; Ebik, Berat; Kaya, Beşir

    2011-03-01

    A case of Fasciola gigantica-induced biliary obstruction and cholestasis is reported in Turkey. The patient was a 37- year-old woman, and suffered from icterus, ascites, and pain in her right upper abdominal region. A total of 7 living adult flukes were recovered during endoscopic retrograde cholangiopancreatography (ERCP). A single dose of triclabendazole was administered to treat possible remaining worms. She was living in a village of southeast of Anatolia region and had sheeps and cows. She had the history of eating lettuce, mallow, dill, and parsley without washing. This is the first case of fascioliasis which was treated via endoscopic biliary extraction during ERCP in Turkey. PMID:21461271

  9. Human distomatosis due to Fasciola hepatica infection in Assam, India.

    PubMed

    Narain, K; Biswas, D; Rajguru, S K; Mahanta, J

    1997-06-01

    A seven year old girl in a rural area of Upper Assam was found to be infected with Fasciola hepatica. The girl gave history of eating watercress. Besides the presence of eggs in the stools, her liver was enlarged and tender. Liver function tests revealed hepato-biliary involvement with increase in serum AKP and ALT. Ultrasonography revealed presence of adult fluke in gall bladder, and a marginal thickening of gall bladder wall. However, the echo pattern of her liver parenchyma was normal. The case is reported for its public health significance and is a maiden report from north eastern India. PMID:9282516

  10. Morphological and Molecular Discrimination of Fasciola Species Isolated From Domestic Ruminants of Urmia City, Iran

    PubMed Central

    YAKHCHALI, Mohammad; MALEKZADEH-VIAYEH, Reza; IMANI-BARAN, Abbas; MARDANI, Karim

    2015-01-01

    Background: The trematodes of the genus Fasciola (the liver flukes) are among the well-known instances of food-borne parasites worldwide. Differentiation of Fasciola species is important because of their different transmission and epidemiological characteristics. The current study was undertaken to discriminate Fasciola species in the domestic ruminants of Urmia city, Iran. Methods: Adult flukes were isolated from the naturally infected livers of the slaughtered water buffaloes and sheep. The flukes were initially identified based on morphological and morphometric parameters. A 618-bp-long fragment of the 28SrRNA gene of Fasciola was amplified by polymerase chain reaction (PCR). The amplified fragment was digested by DraII or AvaII enzymes for a restriction fragment length polymorphism (RFLP) analysis and sequenced for the phylogenetic tree construction. Results: Based on the morphometric examination, the flukes belonged to F. hepatica, F. gigantica and an intermediate Fasciola form. The PCR-RFLP analysis was able to differentiate F. hepatica from F. gigantica. While the phylogenetic reconstruction justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies. Conclusion: To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary. This is crucial for epidemiological surveys and successful clinical management of their infection. PMID:25904945

  11. Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia.

    PubMed

    Hayashi, Kei; Ichikawa-Seki, Madoka; Allamanda, Puttik; Wibowo, Putut Eko; Mohanta, Uday Kumar; Sodirun; Guswanto, Azirwan; Nishikawa, Yoshifumi

    2016-10-01

    Fasciola gigantica and aspermic (hybrid) Fasciola flukes are thought to be distributed in Southeast Asian countries. The objectives of this study were to investigate the distribution of these flukes from unidentified ruminants in western Java, Indonesia, and to determine their distribution history into the area. Sixty Fasciola flukes from western Java were identified as F. gigantica based on the nucleotide sequences of the nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes. The flukes were then analyzed phylogenetically based on the nucleotide sequence of the mitochondrial NADH dehydrogenase subunit 1 (nad1) gene, together with Fasciola flukes from other Asian countries. All but one F. gigantica fluke were classified in F. gigantica haplogroup C, which mainly contains nad1 haplotypes detected in flukes from Thailand, Vietnam, and China. A population genetic analysis suggested that haplogroup C spread from Thailand to the neighboring countries including Indonesia together with domestic ruminants, such as the swamp buffalo, Bubalus bubalis. The swamp buffalo is one of the important definitive hosts of Fasciola flukes in Indonesia, and is considered to have been domesticated in the north of Thailand. The remaining one fluke displayed a novel nad1 haplotype that has never been detected in the reference countries. Therefore, the origin of the fluke could not be established. No hybrid Fasciola flukes were detected in this study, in contrast to neighboring Asian countries. PMID:27266482

  12. Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia.

    PubMed

    Hayashi, Kei; Ichikawa-Seki, Madoka; Allamanda, Puttik; Wibowo, Putut Eko; Mohanta, Uday Kumar; Sodirun; Guswanto, Azirwan; Nishikawa, Yoshifumi

    2016-10-01

    Fasciola gigantica and aspermic (hybrid) Fasciola flukes are thought to be distributed in Southeast Asian countries. The objectives of this study were to investigate the distribution of these flukes from unidentified ruminants in western Java, Indonesia, and to determine their distribution history into the area. Sixty Fasciola flukes from western Java were identified as F. gigantica based on the nucleotide sequences of the nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes. The flukes were then analyzed phylogenetically based on the nucleotide sequence of the mitochondrial NADH dehydrogenase subunit 1 (nad1) gene, together with Fasciola flukes from other Asian countries. All but one F. gigantica fluke were classified in F. gigantica haplogroup C, which mainly contains nad1 haplotypes detected in flukes from Thailand, Vietnam, and China. A population genetic analysis suggested that haplogroup C spread from Thailand to the neighboring countries including Indonesia together with domestic ruminants, such as the swamp buffalo, Bubalus bubalis. The swamp buffalo is one of the important definitive hosts of Fasciola flukes in Indonesia, and is considered to have been domesticated in the north of Thailand. The remaining one fluke displayed a novel nad1 haplotype that has never been detected in the reference countries. Therefore, the origin of the fluke could not be established. No hybrid Fasciola flukes were detected in this study, in contrast to neighboring Asian countries.

  13. The life cycle of Asymphylodora perccotti sp. n. (Trematoda: Lissorchiidae) in the Russian Southern Far East.

    PubMed

    Besprozvannykh, Vladimir V; Ermolenko, Alexey V; Atopkin, Dmitry M

    2012-06-01

    Specimens of Asymphylodora perccotti sp. n. (Trematoda: Lissorchidae) were found in the esophagus of the freshwater fish Perccottus glenii (Odobantidae) taken from the Bolshaya Ussurka River Basin (Primorsky Region, Russian Southern Far East). The first intermediate host of this trematode is a gastropod, Parafossarulus manchouricus, and the secondary hosts are the same mollusk and Boreoelona ussuriensis. Specimens of the new species are similar to A. amnicolae identified by Stunkard in 1959, but the mature worms have larger suckers and shorter ceca. The cercariae of these species are distinguished by body, suckers and pharynx size. These organs in A. perccotti sp. n. are more than one-third larger than what is observed in A. amnicolae. In addition, the new species lacks the capacity for progenesis. Finally, the new species is unusual in that it resides in the fish esophagus instead of the intestine, as is common for most Asymphylodora species. Partial ribosomal DNA sequences and phylogenetic reconstruction sequence data indicate that these worms represent a new digenean species. PMID:22037488

  14. Current Threat of Triclabendazole Resistance in Fasciola hepatica.

    PubMed

    Kelley, Jane M; Elliott, Timothy P; Beddoe, Travis; Anderson, Glenn; Skuce, Philip; Spithill, Terry W

    2016-06-01

    Triclabendazole (TCBZ) is the only chemical that kills early immature and adult Fasciola hepatica (liver fluke) but widespread resistance to the drug greatly compromises fluke control in livestock and humans. The mode of action of TCBZ and mechanism(s) underlying parasite resistance to the drug are not known. Due to the high prevalence of TCBZ resistance (TCBZ-R), effective management of drug resistance is now critical for sustainable livestock production. Here, we discuss the current status of TCBZ-R in F. hepatica, the global distribution of resistance observed in livestock, the possible mechanism(s) of drug action, the proposed mechanisms and genetic basis of resistance, and the prospects for future control of liver fluke infections using an integrated parasite management (IPM) approach.

  15. Serum antibody isotype responses of Fasciola-infected sheep and cattle to excretory and secretory products of Fasciola species.

    PubMed

    Phiri, I K; Phiri, A M; Harrison, L J S

    2006-11-01

    This study investigated the immunoglobulin isotype responses of sheep and cattle chronically infected with Fasciola hepatica and Fasciola gigantica to adult F. hepatica excretory/secretory products (Fh-ES) or F. gigantica excretory/secretory products (Fg-ES), respectively. An antibody enzyme-linked immunosorbent assay (Ab-ELISA) was used to determine serum antibody (total Ig, IgG(1), IgM, IgG(2) and IgA) responses. At necropsy, the mean number of flukes recovered was lower in cattle than in sheep. All F. hepatica and F. gigantica infected sheep and cattle showed an increased total Ig levels from 3 to 4 weeks post-infection (wpi). Among isotypes IgG(1) was most dominant while IgM was the earliest (2 wpi) to be detected in both sheep and cattle infected with both F. hepatica and F. gigantica animals. IgG(2) response was early (2 wpi) in sheep infected by F. hepatica but there was no response in sheep infected with F. gigantica. There was a late and strong IgG(2) response in cattle infected with both flukes. The IgA isotype showed an early and a clear biphasic response in sheep with F. hepatica but was less pronounced in F. gigantica infected sheep. While IgA response to Fh-ES was noticed 5 wpi in F. hepatica infected cattle, it appeared much later (21 wpi) in those infected with F. gigantica. The dominance of IgG(1) isotype in infected sheep and cattle suggest an associated Th2 response. This early response to adult Fasciola spp. ES antigen suggests an early exposure to the antigen presumably through the cross-reacting ES products of juvenile flukes. There is clearly difference in IgG(2) isotype response in cattle (resistant) compared to sheep (susceptible). The late IgG(2) response in cattle may suggest late Th1 involvement in bovine cellular responses to adult Fh-ES/Fg-ES.

  16. Hepatic fibrosis and Fasciola hepatica infection in cattle.

    PubMed

    Marcos, Luis A; Yi, Pedro; Machicado, Alfredo; Andrade, Roy; Samalvides, Frine; Sánchez, Juvenal; Terashima, Angélica

    2007-12-01

    This study focuses on the development of fibrosis of the liver of cattle with Fasciola hepatica infection, correlating with the intensity of infection. Animals with an established diagnosis of chronic F. hepatica infection were identified in a slaughterhouse in Lima, Peru. The study included 24 fresh cattle livers from infected animals and two uninfected controls. Tissues were stored at 4 degrees C for approximately 8 h after which they were brought to a necropsy room and examined. Between 9 and 12 biopsies were randomly obtained from each liver. Histological staining of formalin-fixed liver sections with haematoxylin and eosin (H & E) and Masson's trichrome were performed. Liver samples were examined using a pathology protocol that included 30 items. Histopathologically, 16 out of 30 liver specimens (67.6%) showed diffuse fibrotic lesions (cirrhosis) with a mean number of Fasciola of 116 +/- 30 (range 4-435). Pathological data were matched to number of adult parasites and presence of cirrhosis after being reviewed by two independent pathologists. There was concordance between the two pathologists (K = 0.72). The group with cirrhosis showed an average of 116 +/- 30 adult parasites whereas the group not showing cirrhosis contained 56 +/- 28 flukes (P = 0.2). To measure how number of flukes and diagnosis of cirrhosis are related we used Kendall's tau-b coefficient; the correlation was +0.296 (P = 0.04). Receiver Operating Characteristic (ROC) curve results showed that the best point was 38 parasite adults, which had 93.8% sensitivity and 75% specificity. We conclude that as the number of F. hepatica adult forms increases, the likelihood of developing liver fibrosis will also increase in cattle.

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

    PubMed

    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.

  18. Phenotypic analysis of adults of Fasciola hepatica, Fasciola gigantica and intermediate forms from the endemic region of Gilan, Iran.

    PubMed

    Ashrafi, K; Valero, M A; Panova, M; Periago, M V; Massoud, J; Mas-Coma, S

    2006-12-01

    Fascioliasis is an important human and animal disease caused by Fasciola hepatica and Fasciola gigantica. In Iran, the distribution of these two species overlaps in most areas, including the northern human endemic province of Gilan where both fasciolids are simultaneously found in individual cattle and buffaloes. A phenotypic study of fasciolid adult flukes from naturally infected bovines from Gilan was carried out by means of an exhaustive morphometric analysis using traditional microscopic measurements and an allometric model. The Iranian fasciolids were compared to F. hepatica and F. gigantica standard populations, i.e. from geographical areas where both species do not co-exist (Bolivia and Burkina Faso, respectively). Although morphometric values somewhat overlapped, there were clear differences in allometric growth. The allometric function was adjusted to 25 pairs of variables. Results obtained revealed that Iranian F. hepatica-like specimens are larger than the F. hepatica standard and Iranian F. gigantica-like specimens are longer and narrower than the F. gigantica standard, but with smaller body area. Measurements which permit a specific differentiation in allopatric populations (distance between ventral sucker and posterior end of the body; ratio between body length and body width) overlap in the specimens from Gilan, thus proving the presence of intermediate forms. When compared to the standard populations, the different Iranian fasciolid morphs show greater differences in F. gigantica-like specimens than in F. hepatica-like specimens. This study shows that simple, traditional microscopic measurements may be sufficient for the morphometric characterisation of fasciolids, even in areas where intermediate forms are present. PMID:16901748

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

    PubMed

    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. PMID:25324133

  20. Molecular identification and seasonal infections of species of Fasciola in ruminants from two provinces in China.

    PubMed

    Yuan, W; Liu, J-M; Lu, K; Li, H; Duan, M-M; Feng, J-T; Hong, Y; Liu, Y-P; Zhou, Y; Tong, L-B; Lu, J; Zhu, C-G; Jin, Y-M; Cheng, G-F; Lin, J-J

    2016-05-01

    We determined the prevalence and seasonality of infections by Fasciola of goats and bovine species (cattle and water buffalo) in Hubei and Anhui provinces of China. Faecal samples were collected at 2- to 3-month intervals from 200 goats in Hubei province and from 152 bovine species in Anhui province. All faecal samples were examined for the presence of parasites. We determined the nucleotide sequences of the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA (rDNA) of 39 Fasciola worms from Anhui province. The prevalence of Fasciola infection in goats ranged between 3.5 and 37.0%, with mean eggs per gram (EPG) ranging between 29.0 and 166.0. Prevalence and EPG exhibited downward trends over time with significant differences. The prevalence of Fasciola infection in cattle ranged between 13.3 and 46.2% (mean EPG, 36.4-100.0), and that of water buffalo ranged between 10.3 and 35.4% (mean EPG, 25.0-89.6), with a higher prevalence of infection and EPG from June to October compared with December to March. Analysis of ITS-1 and ITS-2 sequences revealed that F. hepatica and F. gigantica were present in all bovine species of Anhui province and that F. gigantica mainly infected water buffalo. This is the first demonstration of Fasciola infection in Hubei province and detection of F. hepatica and F. gigantica in Anhui province. The present study of Hubei province shows that mass treatment of livestock with closantel sodium injections in April and August/September controlled Fasciola infection effectively.

  1. Partial Hepatectomy for the Resistant Fasciola Hepatica Infection in a Child.

    PubMed

    Belgin, Gülhan; S, Kanık Yüksek; H, Tezer; A, Özkaya Parlakay; B, Dalgıç; A, Dalgıç; G, Yilmaz

    2015-01-01

    Fascioliasis is an emerging and important chronic parasitic disease caused by two trematode liver fluke species: Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica) infecting several herbivorous mammals including cattle, goats, sheep, and humans. We report a 9-year-old girl who suffered from F. hepatica infection and underwent right hepatectomy because of increasing abdominal pain resistant to anthelmintic chemotherapy. When anthelmintic drug treatment is not effective and abdominal pain persists, surgical resection including hepatectomy should be kept in mind for resistant F. hepatica infection.

  2. The effect of immunosuppressants on experimental infection with Fasciola hepatica.

    PubMed

    Corba, J; Spaldonová, R

    1975-01-01

    Results are presented on the effect of immunosuppressive substances such as chlorambucil, cyclophosphamide, azathioprine, amethopterine and a cortizone derivate of betamethasone, on the development of Fasciola hepatica in the rat. The suppression of the immune response of the host to immunosuppressants was reflected in an earlier start of migration of the flukes to the common bile duct, and in an earlier onset of egg production as compared with that in the controls. Of the substances employed, cyclophosphamide and betamethasone were the most effective ones within the period from week 2--6 p.i., which is the time during which the migration of the flukes in the liver parenchyma is highest. Pathological changes in the liver of the animals were less marked than those of the infected controls. Evidence was obtained on an increased pathogenicity of infective larval flukes causing a higher mortality of the hosts in comparison with that of the control animals. On the other hand, the administration of immunosuppressants did neither influence the total number of developed flukes nor the appearance of eosinophilia in the peripheral blood of the treated animals.

  3. Fasciola gigantica thioredoxin glutathione reductase: Biochemical properties and structural modeling.

    PubMed

    Gupta, Ankita; Kesherwani, Manish; Velmurugan, Devadasan; Tripathi, Timir

    2016-08-01

    Platyhelminth thioredoxin glutathione reductase (TGR) is a multifunctional enzyme that crosstalk between the conventional thioredoxin (Trx) and glutathione (GSH) system. It has been validated as a potential drug target in blood flukes. In the present study, we have performed a biochemical study on Fasciola gigantica TGR with substrates DTNB and GSSG. The Michaelis constant (Km) with DTNB was found to be 4.34±0.12μM while it was 61.15±1.50μM with GSSG. The kinetic results were compared with the TGR activities of other helminths. FgTGR showed typical hysteretic behavior with GSSG as other TGRs. We also described a homology-based structure of FgTGR. The cofactors (NADPH and FAD) and substrates (GSSG and DTNB) were docked, and two possible binding sites for substrates were identified in a single chain. The substrates were found to bind more favorably in the second site of TrxR domains. We also presented the first report on binding interaction of DTNB with a TGR. DTNB forms H-bond with His204 and Arg450 of chain A, Sec597, and Gly598 from chain B, salt-bridge with Lys124, and numerous other hydrophobic interactions. Helminth TGR represents an important enzyme in the redox and antioxidant system; hence, its inhibition can be used as an effective strategy against liver flukes. PMID:27112978

  4. Citrate synthase from the liver fluke Fasciola hepatica.

    PubMed

    Zinsser, Veronika L; Moore, Catherine M; Hoey, Elizabeth M; Trudgett, Alan; Timson, David J

    2013-06-01

    Citrate synthase catalyses the first step of the Krebs' tricarboxylic acid cycle. A sequence encoding citrate synthase from the common liver fluke, Fasciola hepatica, has been cloned. The encoded protein sequence is predicted to fold into a largely α-helical protein with high structural similarity to mammalian citrate synthases. Although a hexahistidine-tagged version of the protein could be expressed in Escherichia coli, it was not possible to purify it by nickel-affinity chromatography. Similar results were obtained with a version of the protein which lacks the putative mitochondrial targeting sequence (residues 1 to 29). However, extracts from bacterial cells expressing this version had additional citrate synthase activity after correcting for the endogenous, bacterial activity. The apparent K m for oxaloacetate was found to be 0.22 mM, which is higher than that observed in mammalian citrate synthases. Overall, the sequence and structure of F. hepatica citrate synthase are similar to ones from other eukaryotes, but there are enzymological differences which merit further investigation.

  5. The complete mitochondrial genome of Metorchis orientalis (Trematoda: Opisthorchiidae): Comparison with other closely related species and phylogenetic implications.

    PubMed

    Na, Lu; Gao, Jun-Feng; Liu, Guo-Hua; Fu, Xue; Su, Xin; Yue, Dong-Mei; Gao, Yuan; Zhang, Yan; Wang, Chun-Ren

    2016-04-01

    Metorchis orientalis (Trematoda: Opisthorchiidae) is an important trematode infecting many animals and humans, causing metorchiasis. In the present study, the complete mitochondrial (mt) genome of M. orientalis was sequenced. The complete mt genome of M. orientalis is 13,834 bp circular DNA molecule and contains 12 protein-coding genes, 22 transfer RNA genes, and two ribosomal RNA genes. The gene content and arrangement of M. orientalis is the same as those of Opisthorchiidae trematodes (Opisthorchis viverrini, Opisthorchis felineus and Clonorchis sinensis), but distinct from Schistosoma spp. Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes with three different computational algorithms (Bayesian inference, maximum likelihood and maximum parsimony) revealed that M. orientalis and O. viverrini represent sister taxa. The mt genome provides a novel genetic marker for further studies of the identification, classification and molecular epidemiology of Opisthorchiidae trematodes, and should have implications for the diagnosis, prevention and control of metorchiasis in animals and humans. PMID:26805437

  6. Redescription of Crepidostomum opeongoensis Caira, 1985 (Trematoda: Allocreadiidae) from fish hosts Hiodon alosoides and Hiodon tergisus (Osteichthyes: Hiodontidae)

    USGS Publications Warehouse

    Choudhury, A.; Nelson, P.A.

    2000-01-01

    Crepidostomum opeongoensis Caira, 1985 (Trematoda: Allocreadiidae) is redescribed from fish hosts Hiodon alosoides and Hiodon tergisus in southern Manitoba. The redescription adds details regarding the surface morphology and reproductive structures of the parasite not described previously. These include the characteristic tegumental papillae around the oral opening, paired papillae on the ventral and dorsal surfaces of the body, and tegumental bosses on the dorsal surface of the forebody. Crepidostomum opeongoensis co-occurred with its hypothesized sister species, Crepidostomum illinoiense, in all host individuals harboring the former, and both species are characteristic of hiodontids. The association of C. opeongoensis with hiodontids and the absence of hiodontids in Lake Opeongo where C. opeongoensis was originally reported indicate that the Algonquin Park region may have had hiodontids in the past and that the life cycle may be completed at present without the original fish host.

  7. First phenotypic description of Fasciola hepatica/Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt.

    PubMed

    Periago, M V; Valero, M A; El Sayed, M; Ashrafi, K; El Wakeel, A; Mohamed, M Y; Desquesnes, M; Curtale, F; Mas-Coma, S

    2008-01-01

    Fasciola gigantica is the main fasciolid species in Africa; however, F. hepatica and F. gigantica overlap in some countries. Egypt deserves mentioning because of the emerging situation of human fascioliasis in the Nile Delta area. The morphometric characteristics of fasciolid adults infecting the main livestock species present in the Nile Delta human endemic area are analyzed through a computer image analysis system (CIAS) on the basis of standardized measurements known to be useful for the differentiation of both fasciolid species. This is the first time that such a study is performed in an African country and, therefore, the results are compared to (i) F. hepatica (European Mediterranean area) and F. gigantica (Burkina Faso) standard populations, i.e. geographical areas where both species do not co-exist, and (ii) F. hepatica and F. gigantica populations from geographical areas where both species do co-exist, including the presence of intermediate forms (Iran). Results indicate the presence of F. hepatica, F. gigantica and intermediate forms (Fasciola sp.) in Egypt for the first time, and demonstrate the usefulness of CIAS for the phenotypic characterization of liver fluke adults from a concrete fascioliasis endemic area. Body roundness, body length over body width, and distance between the ventral sucker and the posterior end of the body provide useful tools for studying inter- and intraspecific morphological diversity in Fasciola adults. The application of these markers to specimens from geographical areas where F. hepatica and F. gigantica co-exist, such as in Egypt and Iran, suggest a strong population-level variation in Fasciola adult morphology. PMID:18006385

  8. Cross-reactivity of Schistosoma mansoni-Fasciola gigantica influenced by saponins.

    PubMed

    Maghraby, Amany Sayed; Shaker, Kamel H; Gaber, Hanaa M

    2009-01-01

    The aim of the present work was to investigate the Schistosoma mansoni and Fasciola gigantica cross-reactivity between adult worms and egg homogenates of the parasites. Immunoprophylactic effects of crude Schistosoma mansoni worms and egg antigens mixed with or without saponins extracted from Atriplex nummularia were studied followed by challenge with 80 cercariae of Schistosoma mansoni. Our results showed that post 1st immunization with schistosome egg antigens (SEA) there was a significant change (P approximately 0.05) in the IgM levels against Fasciola egg homogenate (FgEH) without saponins. Post 2nd immunization with SEA mixed with saponins the levels of IgM increased significantly (P approximately 0.05) against Fasciola worm homogenate (FgWH) as compared with a non-immunized group. Post 2nd immunization the level of IgG was significantly elevated (P approximately 0.05) by SEA mixed with saponins against FgWH. Post 2nd immunizations with SEA mixed with saponins showed a significant change (P approximately 0.05) in IgG levels against FgEH. These results clearly demonstrated that there is a cross-reactivity between Schistosoma mansoni eggs and Fasciola gigantica worms and eggs. Saponins were found to be immunostimulatory adjuvants in our study. PMID:19526726

  9. Fasciola hepatica from naturally infected sheep and cattle in Great Britain are diploid.

    PubMed

    Beesley, N J; Cwiklinski, K; Williams, D J L; Hodgkinson, J

    2015-08-01

    Diploid (2n = 2x = 20) and triploid (2n = 3x = 30) Fasciola hepatica have been reported in the UK, and in Asia diploid, triploid and mixoploid (2x/3x) Fasciola spp. exist but there is little information to indicate how common triploidy is, particularly in UK fluke. Here the ploidy of 565 adult F. hepatica from 66 naturally infected British sheep and 150 adult F. hepatica from 35 naturally infected British cattle was determined. All 715 of these parasites were diploid, based on observation of 10 bivalent chromosomes and sperm (n = 335) or, since triploids are aspermic, sperm alone (n = 380). This constitutes the first extensive analysis of the ploidy of F. hepatica field isolates from Great Britain and shows that most F. hepatica isolated from cattle and sheep are diploid and have the capacity to sexually reproduce. These data suggest that triploidy, and by extension parthenogenesis, is rare or non-existent in wild British F. hepatica populations. Given that F. hepatica is the only species of Fasciola present in Britain our results indicate that the parasite is predominantly diploid in areas where F. hepatica exists in isolation and suggests that triploidy may only originate in natural populations where co-infection of F. hepatica and its sister species Fasciola gigantica commonly occurs.

  10. Inhibition Effect of pH on the Hatchability of Fasciola Miracidia under Laboratory Conditions

    PubMed Central

    YAKHCHALI, Mohammad; BAHRAMNEJAD, Kia

    2016-01-01

    Background: Fasciolosis, caused by the liver flukes of the genus Fasciola, is one of the most prevalent diseases of domestic livestock and human throughout the world, imposing considerable economic losses. The present study was aimed to assess the effects of different pH values on hatching rate of Fasciola miracidia. Methods: The flukes were isolated from the infected livers of the slaughtered ruminants at the abattoir of Urmia City, Iran, crushed thoroughly and sieved for isolation of the Fasciola eggs. The eggs were washed up several times by PBS (0.01N, pH 7.2). They were incubated at different pH values of 7±0.1 (control) and 3–9.5 (treatments) at 28°C for 16 days. Results: The maximum hatching rate was observed at pH 7 (14.93±0.65%), while no miracidia were hatched at pH 3 and/or pH 9–9.5. There were significant differences between the hatching rate of the treatments and that of the control group. Conclusion: Water pH is proven to be a crucial factor affecting the life cycle of Fasciola and its epidemiology. PMID:27095966

  11. Urea cycle of Fasciola gigantica: purification and characterization of arginase.

    PubMed

    Mohamed, Saleh A; Fahmy, Afaf S; Mohamed, Tarek M; Hamdy, Soha M

    2005-11-01

    The ornithine-urea cycle has been investigated in Fasciola gigantica. Agrinase had very high activity compared to the other enzymes. Carbamoyl phosphate synthetase and ornithine carbamoyltransferase had very low activity. A moderate enzymatic activity was recorded for argininosuccinate synthetase and argininosuccinate lyase. The low levels of F. gigantica urea cycle enzymes except to the arginase suggest the urea cycle is operative but its role is of a minor important. The high level of arginase activity may benefit for the hydrolysis of the exogenous arginine to ornithine and urea. Two arginases Arg I and Arg II were separated by DEAE-Sepharose column. Further purification was restricted to Arg II with highest activity. The molecular weight of Arg II, as determined by gel filtration and SDS-PAGE, was 92,000. The enzyme was capable to hydrolyze l-arginine and to less extent l-canavanine at arginase:canavanase ratio (>10). The enzyme exhibited a maximal activity at pH 9.5 and Km of 6 mM. The optimum temperature of F. gigantica Arg II was 40 degrees C and the enzyme was stable up to 30 degrees C and retained 80% of its activity after incubation at 40 degrees C for 15 min and lost all of its activity at 50 degrees C. The order of effectiveness of amino acids as inhibitors of enzyme was found to be lysine>isoleucine>ornithine>valine>leucine>proline with 67%, 43%, 31%, 25%, 23% and 15% inhibition, respectively. The enzyme was activated with Mn2+, where the other metals Fe2+, Ca2+, Hg2+, Ni2+, Co2+ and Mg2+ had inhibitory effects. PMID:16125991

  12. Climate-based risk models for Fasciola hepatica in Colombia.

    PubMed

    Valencia-López, Natalia; Malone, John B; Carmona, Catalina Gómez; Velásquez, Luz E

    2012-09-01

    A predictive Fasciola hepatica model, based on the growing degree day-water budget (GDD-WB) concept and the known biological requirements of the parasite, was developed within a geographical information system (GIS) in Colombia. Climate-based forecast index (CFI) values were calculated and represented in a national-scale, climate grid (18 x 18 km) using ArcGIS 9.3. A mask overlay was used to exclude unsuitable areas where mean annual temperature exceeded 25 °C, the upper threshold for development and propagation of the F. hepatica life cycle. The model was then validated and further developed by studies limited to one department in northwest Colombia. F. hepatica prevalence data was obtained from a 2008-2010 survey in 10 municipalities of 6,016 dairy cattle at 673 herd study sites, for which global positioning system coordinates were recorded. The CFI map results were compared to F. hepatica environmental risk models for the survey data points that had over 5% prevalence (231 of the 673 sites) at the 1 km2 scale using two independent approaches: (i) a GIS map query based on satellite data parameters including elevation, enhanced vegetation index and land surface temperature day-night difference; and (ii) an ecological niche model (MaxEnt), for which geographic point coordinates of F. hepatica survey farms were used with BioClim data as environmental variables to develop a probability map. The predicted risk pattern of both approaches was similar to that seen in the forecast index grid. The temporal risk, evaluated by the monthly CFIs and a daily GDD-WB forecast software for 2007 and 2008, revealed a major July-August to January transmission period with considerable inter-annual differences.

  13. Fasciola hepatica: the redial generations in juvenile Pseudosuccinea columella.

    PubMed

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

    2015-08-01

    Juvenile Pseudosuccinea columella, measuring 1 or 2 mm in height, were subjected to single-miracidium infections with Fasciola hepatica to determine the developmental pattern of redial generations and count free and live rediae according to their generation. Controls were constituted of juvenile Galba truncatula infected according to the same protocol. In the four groups, redial counts were performed in snails dissected every week from day 7 to day 49 post-exposure at 20 °C. Most infected snails showed a normal development of redial generations, whatever the lymnaeid species. In P. columella, the total number of live rediae on day 49 was 24.6 and 34.6 per infected snail in the 1 and 2-mm groups, respectively (instead of 11.5 and 18.8 rediae in the corresponding groups of G. truncatula). A single mother redia (R1a) producing only daughter rediae of the second generation was noted in each snail of three groups, while the 2-mm P. columella showed the presence of a single (43 snails/71) or two (28/71) R1a redia(e) within their bodies. The mean number of other mother rediae and that of daughter rediae of the second generation were low in the 1 and 2-mm groups of both lymnaeids. Besides, there was a delay in redial development. The development of two live R1a rediae in several P. columella from the 2-mm group needs to verify if this process would be specific to P. columella or would occur in other lymnaeid species known for their good susceptibility to the digenean and their larger size to allow harbouring rediae. PMID:25920445

  14. Controversial aspects of the life cycle of Fasciola hepatica.

    PubMed

    Moazeni, Mohammad; Ahmadi, Amin

    2016-10-01

    Fasciola hepatica is a well-known helminth parasite, with significant economic and public health importance all over the world. It has been known since more than 630 years ago and a considerable research work has been carried out on the life cycle of this important parasite. In the hepatic phase of the life cycle of F. hepatica, it is assumed that the young flukes, after about 6-7 weeks of migration in the liver parenchyma, enter into the bile ducts of the definitive hosts and become sexually mature. Even though the secretion of cysteine peptidases including cathepsin L and B proteases by F. hepatica may justify this opinion, because of several scientific reasons and based on the experimental studies conducted in different animals (reviewed in this article), the entry of parasites into the bile ducts, after their migration in the liver parenchyma seems to be doubtful. However, considering all the facts relating to the hepatic and biliary phases of the life cycle of F. hepatica, two alternative ideas are suggested: 1) some of the migrating juvenile flukes may enter into the bile ducts immediately after reaching the liver parenchyma while they are still very small, or 2) when newly excysted juvenile flukes are penetrating into the intestinal wall to reach the liver through the abdominal cavity, a number of these flukes may enter into the choleduct and reach the hepatic bile ducts, where they mature. According to the previously performed natural and experimental studies in different animals and human beings, the supporting and opposing evidences for the current opinion as well as the evidences that might justify the two new ideas are reviewed and discussed briefly. In conclusion, our present knowledge about the time and quality of the entry of F. hepaticas into the bile ducts, seems to be insufficient, therefore, there are still some dark corners and unknown aspects in this field that should be clarified.

  15. Predicting impacts of climate change on Fasciola hepatica risk.

    PubMed

    Fox, Naomi J; White, Piran C L; McClean, Colin J; Marion, Glenn; Evans, Andy; Hutchings, Michael R

    2011-01-01

    Fasciola hepatica (liver fluke) is a physically and economically devastating parasitic trematode whose rise in recent years has been attributed to climate change. Climate has an impact on the free-living stages of the parasite and its intermediate host Lymnaea truncatula, with the interactions between rainfall and temperature having the greatest influence on transmission efficacy. There have been a number of short term climate driven forecasts developed to predict the following season's infection risk, with the Ollerenshaw index being the most widely used. Through the synthesis of a modified Ollerenshaw index with the UKCP09 fine scale climate projection data we have developed long term seasonal risk forecasts up to 2070 at a 25 km square resolution. Additionally UKCIP gridded datasets at 5 km square resolution from 1970-2006 were used to highlight the climate-driven increase to date. The maps show unprecedented levels of future fasciolosis risk in parts of the UK, with risk of serious epidemics in Wales by 2050. The seasonal risk maps demonstrate the possible change in the timing of disease outbreaks due to increased risk from overwintering larvae. Despite an overall long term increase in all regions of the UK, spatio-temporal variation in risk levels is expected. Infection risk will reduce in some areas and fluctuate greatly in others with a predicted decrease in summer infection for parts of the UK due to restricted water availability. This forecast is the first approximation of the potential impacts of climate change on fasciolosis risk in the UK. It can be used as a basis for indicating where active disease surveillance should be targeted and where the development of improved mitigation or adaptation measures is likely to bring the greatest benefits. PMID:21249228

  16. Primary experimental infection of riverine buffaloes with Fasciola gigantica.

    PubMed

    Yadav, S C; Sharma, R L; Kalicharan, A; Mehra, U R; Dass, R S; Verma, A K

    1999-05-01

    The clinical course of the primary experimental Fasciola gigantica infection was investigated in riverine buffalo calves of the Murrah breed. Nine male calves aged 12-15 months were randomly assigned to two groups of five (Group I) and four (Group II) animals. Each animal in Group I, was orally infected with 1000 metacercariae (mc) of F. gigantica, whereas Group II animals did not receive any infection dose and served as uninfected controls. No clinical signs of fasciolosis were observed until the sixth week post-infection (PI). Group I animals, however, developed recognised symptoms of acute fasciolosis, comprising apyrexic inappetance, anemia, poor weight gain, diarrhoea and sub-mandibular and facial oedema, respectively, from 5, 6, 8, 16 and 17 weeks PI. The signs were intermittent in nature and of variable duration. The prepatent period was of 92-97 days (mean 95.2 +/- 3.1). One of the five infected animals died on Day 147 PI. At necropsy, 36.8 +/- 11.0% of the infection dose was recovered as adult fluke population. The gross lesions were primarily biliary in nature. Group II, the uninfected controls, throughout the study period of 165 days PI, did not show any symptom and were negative for F. gigantica. The study demonstrated that the onset of adverse effects of F. gigantica on the growth and health of the infected host was mainly noted during late prepatency much before coprological prediction and diagnosis. The significance of preventive therapy against fasciolosis during prepatency has been stressed in endemic areas. PMID:10384904

  17. Transcriptomic Study on Ovine Immune Responses to Fasciola hepatica Infection

    PubMed Central

    Fu, Yan; Chryssafidis, Andreas L.; Browne, John A.; O'Sullivan, Jack; McGettigan, Paul A.; Mulcahy, Grace

    2016-01-01

    Background Fasciola hepatica is not only responsible for major economic losses in livestock farming, but is also a major food-borne zoonotic agent, with 180 million people being at risk of infection worldwide. This parasite is sophisticated in manipulating the hosts’ immune system to benefit its own survival. A better understanding of the mechanisms underpinning this immunomodulation is crucial for the development of control strategies such as vaccines. Methodology/principal findings This in vivo study investigated the global gene expression changes of ovine peripheral blood mononuclear cells (PBMC) response to both acute & chronic infection of F. hepatica, and revealed 6490 and 2364 differential expressed genes (DEGS), respectively. Several transcriptional regulators were predicted to be significantly inhibited (e.g. IL12 and IL18) or activated (e.g. miR155-5p) in PBMC during infection. Ingenuity Pathway Analysis highlighted a series of immune-associated pathways involved in the response to infection, including ‘Transforming Growth Factor Beta (TGFβ) signaling’, ‘Production of Nitric Oxide in Macrophages’, ‘Toll-like Receptor (TLRs) Signaling’, ‘Death Receptor Signaling’ and ‘IL17 Signaling’. We hypothesize that activation of pathways relevant to fibrosis in ovine chronic infection, may differ from those seen in cattle. Potential mechanisms behind immunomodulation in F. hepatica infection are a discussed. Significance In conclusion, the present study performed global transcriptomic analysis of ovine PBMC, the primary innate/adaptive immune cells, in response to infection with F. hepatica, using deep-sequencing (RNAseq). This dataset provides novel information pertinent to understanding of the pathological processes in fasciolosis, as well as a base from which to further refine development of vaccines. PMID:27661612

  18. Controversial aspects of the life cycle of Fasciola hepatica.

    PubMed

    Moazeni, Mohammad; Ahmadi, Amin

    2016-10-01

    Fasciola hepatica is a well-known helminth parasite, with significant economic and public health importance all over the world. It has been known since more than 630 years ago and a considerable research work has been carried out on the life cycle of this important parasite. In the hepatic phase of the life cycle of F. hepatica, it is assumed that the young flukes, after about 6-7 weeks of migration in the liver parenchyma, enter into the bile ducts of the definitive hosts and become sexually mature. Even though the secretion of cysteine peptidases including cathepsin L and B proteases by F. hepatica may justify this opinion, because of several scientific reasons and based on the experimental studies conducted in different animals (reviewed in this article), the entry of parasites into the bile ducts, after their migration in the liver parenchyma seems to be doubtful. However, considering all the facts relating to the hepatic and biliary phases of the life cycle of F. hepatica, two alternative ideas are suggested: 1) some of the migrating juvenile flukes may enter into the bile ducts immediately after reaching the liver parenchyma while they are still very small, or 2) when newly excysted juvenile flukes are penetrating into the intestinal wall to reach the liver through the abdominal cavity, a number of these flukes may enter into the choleduct and reach the hepatic bile ducts, where they mature. According to the previously performed natural and experimental studies in different animals and human beings, the supporting and opposing evidences for the current opinion as well as the evidences that might justify the two new ideas are reviewed and discussed briefly. In conclusion, our present knowledge about the time and quality of the entry of F. hepaticas into the bile ducts, seems to be insufficient, therefore, there are still some dark corners and unknown aspects in this field that should be clarified. PMID:27475124

  19. Cytosolic superoxide dismutase can provide protection against Fasciola gigantica.

    PubMed

    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. PMID:27338185

  20. Characterisation of Fasciola hepatica cytochrome c peroxidase as an enzyme with potential antioxidant activity in vitro.

    PubMed

    Campos, E G; Hermes-Lima, M; Smith, J M; Prichard, R K

    1999-05-01

    Cytochrome c peroxidase oxidises hydrogen peroxide using cytochrome c as the electron donor. This enzyme is found in yeast and bacteria and has been also described in the trematodes Fasciola hepatica and Schistosoma mansoni. Using partially purified cytochrome c peroxidase samples from Fasciola hepatica we evaluated its role as an antioxidant enzyme via the investigation of its ability to protect against oxidative damage to deoxyribose in vitro. A system containing FeIII-EDTA plus ascorbate was used to generate reactive oxygen species superoxide radical, H2O2 as well as the hydroxyl radical. Fasciola hepatica cytochrome c peroxidase effectively protected deoxyribose against oxidative damage in the presence of its substrate cytochrome c. This protection was proportional to the amount of enzyme added and occurred only in the presence of cytochrome c. Due to the low specific activity of the final partially purified sample the effects of ascorbate and calcium chloride on cytochrome c peroxidase were investigated. The activity of the partially purified enzyme was found to increase between 10 and 37% upon reduction with ascorbate. However, incubation of the partially purified enzyme with 1 mM calcium chloride did not have any effect on enzyme activity. Our results showed that Fasciola hepatica CcP can protect deoxyribose from oxidative damage in vitro by blocking the formation of the highly toxic hydroxyl radical (.OH). We suggest that the capacity of CcP to inhibit .OH-formation, by efficiently removing H2O2 from the in vitro oxidative system, may extend the biological role of CcP in response to oxidative stress in Fasciola hepatica. PMID:10404259

  1. Monitoring of Fasciola Species Contamination in Water Dropwort by cox1 Mitochondrial and ITS-2 rDNA Sequencing Analysis.

    PubMed

    Choi, In-Wook; Kim, Hwang-Yong; Quan, Juan-Hua; Ryu, Jae-Gee; Sun, Rubing; Lee, Young-Ha

    2015-10-01

    Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.

  2. Monitoring of Fasciola Species Contamination in Water Dropwort by cox1 Mitochondrial and ITS-2 rDNA Sequencing Analysis

    PubMed Central

    Choi, In-Wook; Kim, Hwang-Yong; Quan, Juan-Hua; Ryu, Jae-Gee; Sun, Rubing; Lee, Young-Ha

    2015-01-01

    Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples. PMID:26537044

  3. Efficacy of a single oral dose of oxfendazole against Fasciola hepatica in naturally infected sheep.

    PubMed

    Gomez-Puerta, Luis A; Gavidia, Cesar; Lopez-Urbina, Maria T; Garcia, Hector H; Gonzalez, Armando E

    2012-03-01

    The efficacy of a single oral dose of 30 mg/kg of oxfendazole against Fasciola hepatica was evaluated in a controlled study in naturally infected sheep. Sheep were diagnosed by stool microscopy after sedimentation, and positive animals were randomized to oxfendazole (N = 20) or no treatment (N = 20). A new stool exam was performed 10 days after treatment. All stool microscopies were performed masked to the treatment group. No side effects were noticed. All sheep in the control group remained infected with similar counts of eggs per gram of stools. None of the animals in the treatment group showed Fasciola eggs in stools after 10 days of treatment. A single dose of oxfendazole is highly effective against F. hepatica, providing a new drug alternative for the control of fascioliasis or integrated zoonosis control.

  4. Efficacy of a Single Oral Dose of Oxfendazole against Fasciola hepatica in Naturally Infected Sheep

    PubMed Central

    Gomez-Puerta, Luis A.; Gavidia, Cesar; Lopez-Urbina, Maria T.; Garcia, Hector H.; Gonzalez, Armando E.

    2012-01-01

    The efficacy of a single oral dose of 30 mg/kg of oxfendazole against Fasciola hepatica was evaluated in a controlled study in naturally infected sheep. Sheep were diagnosed by stool microscopy after sedimentation, and positive animals were randomized to oxfendazole (N = 20) or no treatment (N = 20). A new stool exam was performed 10 days after treatment. All stool microscopies were performed masked to the treatment group. No side effects were noticed. All sheep in the control group remained infected with similar counts of eggs per gram of stools. None of the animals in the treatment group showed Fasciola eggs in stools after 10 days of treatment. A single dose of oxfendazole is highly effective against F. hepatica, providing a new drug alternative for the control of fascioliasis or integrated zoonosis control. PMID:22403323

  5. In vitro tegumental alterations on adult Fasciola gigantica caused by mefloquine.

    PubMed

    Shalaby, Hatem A; El Namaky, Amira H; Kamel, Reem O A

    2016-03-01

    Emergence of drug-resistant Fasciola strains has drawn the attention of many authors to alternative drugs. The purpose of this study is to explore the in vitro effect of the antimalarial mefloquine against adult Fasciola gigantica. Light and scanning electron microscopic observations could be used to determine the target of the drug following 6 and 12 h of incubation in medium containing mefloquine at three different concentrations 10, 20 and 30 μg/mL, as morphological changes could be observed. These changes occurred in definite sequences in response to mefloquine, and were consisted of swelling, vacuolization that was later disrupted, leading to desquamation of the tegument, resulting in exposure and disruption of basal lamina and the dislodging of spines. It is concluded that mefloquine presented itself as a drug that might become important in trematode chemotherapy, with the tegument being an important drug target. PMID:27065615

  6. [A clinical presentation of a very rare infection: parenchymal Fasciola hepatica].

    PubMed

    Sapmaz, Ferdane; Kalkan, Ismail Hakkı; Guliter, Sefa; Nazlıoğlu, Adem

    2013-01-01

    Fascioliasis is primarily an infection of livestock such as cattle and sheep, caused by the flat, brown liver fluke Fasciola hepatica. Humans are accidental hosts. The diagnosis of infection depends on suspicion. Radiologic findings are specific. Usually, Computed Tomography (CT) and other imaging studies show hypodense migratory lesions of the liver. The development of a chronic liver abscess appears to be extremely rare. Here we present our case with hepatic abscess due to F. hepatica, which is a rare clinical presentation.

  7. Genetic analysis of Fasciola isolates from cattle in Korea based on second internal transcribed spacer (ITS-2) sequence of nuclear ribosomal DNA.

    PubMed

    Choe, Se-Eun; Nguyen, Thuy Thi-Dieu; Kang, Tae-Gyu; Kweon, Chang-Hee; Kang, Seung-Won

    2011-09-01

    Nuclear ribosomal DNA sequence of the second internal transcribed spacer (ITS-2) has been used efficiently to identify the liver fluke species collected from different hosts and various geographic regions. ITS-2 sequences of 19 Fasciola samples collected from Korean native cattle were determined and compared. Sequence comparison including ITS-2 sequences of isolates from this study and reference sequences from Fasciola hepatica and Fasciola gigantica and intermediate Fasciola in Genbank revealed seven identical variable sites of investigated isolates. Among 19 samples, 12 individuals had ITS-2 sequences completely identical to that of pure F. hepatica, five possessed the sequences identical to F. gigantica type, whereas two shared the sequence of both F. hepatica and F. gigantica. No variations in length and nucleotide composition of ITS-2 sequence were observed within isolates that belonged to F. hepatica or F. gigantica. At the position of 218, five Fasciola containing a single-base substitution (C>T) formed a distinct branch inside the F. gigantica-type group which was similar to those of Asian-origin isolates. The phylogenetic tree of the Fasciola spp. based on complete ITS-2 sequences from this study and other representative isolates in different locations clearly showed that pure F. hepatica, F. gigantica type and intermediate Fasciola were observed. The result also provided additional genetic evidence for the existence of three forms of Fasciola isolated from native cattle in Korea by genetic approach using ITS-2 sequence. PMID:21494845

  8. Fascioliasis of livestock and snail host for Fasciola in the Altiplano Region of Bolivia.

    PubMed

    Ueno, H; Arandia, R; Morales, G; Medina, G

    1975-01-01

    Fascioliasis caused by Fasciola hepatica was a serious problem for sheep and alpacas in the Altiplano Region of Bolivia. In some provinces close to Lake Titicaca, the raising of sheep was forced to discontinue, because infection with the fluke made it unprofitable and almost impossible. It was proved that in the Altiplano Region, two species of freshwater snails, Lymnaea viatrix and L. cubensis var., served as intermediate hosts for F. hepatica. In some subtropical areas of Bolivia, these snails could not be found, although other Lymnaea sp. was widely distributed there. As it is possible for Lymnaea sp. to be intermediate host for the fluke, further studies are required on the identification. Acute fascioliasis of sheep occurred in the Altiplano Region principally during a period from May to July, or the dry season. In some areas, the mortality rate of infected sheep was roughly estimated as 15 to 25% annually. Contamination with Fasciola metacercariae of herbage and semi-aquatic plants grown in a swamp in one of these areas was biologically assessed, using guinea pigs. Plants of Compositae and Eleocharis sp. were contaminated most intensely and those of Senicio sp. and Vallisneria sp. carried a fairly large number of cysts, while plants of Scirpus sp. and Ranunclaceae carried only a few cysts. No signs of Fasciola infection were observed in any animal given the plants of Liliaceae.

  9. An optimized DNA extraction and multiplex PCR for the detection of Fasciola sp. in lymnaeid snails.

    PubMed

    Caron, Y; Righi, S; Lempereur, L; Saegerman, C; Losson, B

    2011-05-31

    This study deals with the development and validation of an original PCR protocol to assess the presence of Fasciola hepatica in Galba truncatula its main intermediate host in Western Europe. In the present study two DNA extraction techniques are compared and a new multiplex PCR is described. The Chelex(®) DNA extraction technique showed to be more appropriate than the classical Phenol/Chloroform/Proteinase K based method because of the absence of toxic organic solvent, shorter duration and lower cost, and a higher reproducibility regarding DNA concentrations and wavelength ratios. The multiplex PCR was set up to amplify the lymnaeid internal transcribed spacer 2 sequence (500-600 bp) that act as an internal control and a 124 bp Fasciola sp. sequence that is repeated more than 300,000 times in fluke whole genome. Ninety six snails were pooled and 6 snails (6.25%) found positive for Fasciola sp. The limit of detection is lower than the minimal biological infestation unit (one miracidium). DNA extracts from Paramphistomum daubneyi, Dicrocoelium lanceolatum, and Fascioloides magna did not cross react. PMID:21242033

  10. Inhibition of acetylcholinesterase and cytochrome oxidase activity in Fasciola gigantica cercaria by phytoconstituents.

    PubMed

    Sunita, Kumari; Habib, Maria; Kumar, P; Singh, Vinay Kumar; Husain, Syed Akhtar; Singh, D K

    2016-02-01

    Fasciolosis is an important cattle and human disease caused by Fasciola hepatica and Fasciola gigantica. One of the possible methods to control this problem is to interrupt the life cycle of Fasciola by killing its larva (redia and cercaria) in host snail. Molecular identification of cercaria larva of F. gigantica was done by comparing the nucleotide sequencing with adult F. gigantica. It was noted that nucleotide sequencing of cercaria larva and adult F. gigantica were 99% same. Every month during the year 2011-2012, in vivo treatment with 60% of 4 h LC50 of phyto cercaricides citral, ferulic acid, umbelliferone, azadirachtin and allicin caused significant inhibition of acetylcholinesterase (AChE) and cytochrome oxidase activity in the treated cercaria larva of F. gigantica. Whereas, activity of both enzymes were not significantly altered in the nervous tissues of vector snail Lymnaea acuminata exposed to same treatments. Maximum reduction in AChE (1.35% of control in month of June) and cytochrome oxidase (3.71% of control in the month of July) activity were noted in the cercaria exposed to 60% of 4 h LC50 of azadirachtin and allicin, respectively. PMID:26536397

  11. Fascioliasis Control: In Vivo and In Vitro Phytotherapy of Vector Snail to Kill Fasciola Larva

    PubMed Central

    Sunita, Kumari; Singh, D. K.

    2011-01-01

    Snail is one of the important components of an aquatic ecosystem, it acts as intermediate host of Fasciola species. Control of snail population below a certain threshold level is one of the important methods in the campaign to reduce the incidence of fascioliasis. Life cycle of the parasite can be interrupted by killing the snail or Fasciola larva redia and cercaria in the snail body. In vivo and in vitro toxicity of the plant products and their active component such as citral, ferulic acid, umbelliferone, azadirachtin, and allicin against larva of Fasciola in infected snail Lymnaea acuminata were tested. Mortality of larvae were observed at 2 h, 4 h, 6 h, and 8 h, of treatment. In in vivo treatment, azadirachtin caused highest mortality in redia and cercaria larva (8 h, LC50 0.11, and 0.05 mg/L) whereas in in vitro condition allicin was highly toxic against redia and cercaria (8 h, LC50 0.01, and 0.009 mg/L). Toxicity of citral was lowest against redia and cercaria larva. PMID:22132306

  12. CIAS detection of Fasciola hepatica/F. gigantica intermediate forms in bovines from Bangladesh.

    PubMed

    Ahasan, Syed Ali; Valero, M Adela; Chowdhury, Emdadul Haque; Islam, Mohammad Taohidul; Islam, Mohammad Rafiqul; Hussain Mondal, Mohammad Motahar; Peixoto, Raquel V; Berinde, Lavinia; Panova, Miroslava; Mas-Coma, Santiago

    2016-03-01

    Fascioliasis is an important food-borne parasitic zoonosis caused by two trematode species, Fasciola hepatica and Fasciola gigantica. The characterisation and differentiation of Fasciola populations is crucial to control the disease, given the different transmission, epidemiology and pathology characteristics of the two species. Lineal biometric features of adult liver flukes infecting livestock have been studied to characterise and discriminate fasciolids from Bangladesh. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected bovines (cattle, buffaloes) throughout the country. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardised measurements and the logistic model of the body growth and development of fasciolids in the different host groups. Since it is the first ever comprehensive study of this kind undertaken in Bangladesh, the results are compared to pure fasciolid populations of F. hepatica from the European Mediterranean area and F. gigantica from Burkina Faso, geographical areas where both species do not co-exist. Principal component analysis showed that the biometric characteristics of fasciolids from Bangladesh are situated between F. hepatica and F. gigantica standard populations, indicating the presence of phenotypes of intermediate forms in Bangladesh. These results are analysed by considering the present emergence of animal fascioliasis, the local lymnaeid fauna, the impact of climate change, and the risk of human infection in the country.

  13. CIAS detection of Fasciola hepatica/F. gigantica intermediate forms in bovines from Bangladesh.

    PubMed

    Ahasan, Syed Ali; Valero, M Adela; Chowdhury, Emdadul Haque; Islam, Mohammad Taohidul; Islam, Mohammad Rafiqul; Hussain Mondal, Mohammad Motahar; Peixoto, Raquel V; Berinde, Lavinia; Panova, Miroslava; Mas-Coma, Santiago

    2016-03-01

    Fascioliasis is an important food-borne parasitic zoonosis caused by two trematode species, Fasciola hepatica and Fasciola gigantica. The characterisation and differentiation of Fasciola populations is crucial to control the disease, given the different transmission, epidemiology and pathology characteristics of the two species. Lineal biometric features of adult liver flukes infecting livestock have been studied to characterise and discriminate fasciolids from Bangladesh. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected bovines (cattle, buffaloes) throughout the country. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardised measurements and the logistic model of the body growth and development of fasciolids in the different host groups. Since it is the first ever comprehensive study of this kind undertaken in Bangladesh, the results are compared to pure fasciolid populations of F. hepatica from the European Mediterranean area and F. gigantica from Burkina Faso, geographical areas where both species do not co-exist. Principal component analysis showed that the biometric characteristics of fasciolids from Bangladesh are situated between F. hepatica and F. gigantica standard populations, indicating the presence of phenotypes of intermediate forms in Bangladesh. These results are analysed by considering the present emergence of animal fascioliasis, the local lymnaeid fauna, the impact of climate change, and the risk of human infection in the country. PMID:27078650

  14. An optimized DNA extraction and multiplex PCR for the detection of Fasciola sp. in lymnaeid snails.

    PubMed

    Caron, Y; Righi, S; Lempereur, L; Saegerman, C; Losson, B

    2011-05-31

    This study deals with the development and validation of an original PCR protocol to assess the presence of Fasciola hepatica in Galba truncatula its main intermediate host in Western Europe. In the present study two DNA extraction techniques are compared and a new multiplex PCR is described. The Chelex(®) DNA extraction technique showed to be more appropriate than the classical Phenol/Chloroform/Proteinase K based method because of the absence of toxic organic solvent, shorter duration and lower cost, and a higher reproducibility regarding DNA concentrations and wavelength ratios. The multiplex PCR was set up to amplify the lymnaeid internal transcribed spacer 2 sequence (500-600 bp) that act as an internal control and a 124 bp Fasciola sp. sequence that is repeated more than 300,000 times in fluke whole genome. Ninety six snails were pooled and 6 snails (6.25%) found positive for Fasciola sp. The limit of detection is lower than the minimal biological infestation unit (one miracidium). DNA extracts from Paramphistomum daubneyi, Dicrocoelium lanceolatum, and Fascioloides magna did not cross react.

  15. Inhibition of acetylcholinesterase and cytochrome oxidase activity in Fasciola gigantica cercaria by phytoconstituents.

    PubMed

    Sunita, Kumari; Habib, Maria; Kumar, P; Singh, Vinay Kumar; Husain, Syed Akhtar; Singh, D K

    2016-02-01

    Fasciolosis is an important cattle and human disease caused by Fasciola hepatica and Fasciola gigantica. One of the possible methods to control this problem is to interrupt the life cycle of Fasciola by killing its larva (redia and cercaria) in host snail. Molecular identification of cercaria larva of F. gigantica was done by comparing the nucleotide sequencing with adult F. gigantica. It was noted that nucleotide sequencing of cercaria larva and adult F. gigantica were 99% same. Every month during the year 2011-2012, in vivo treatment with 60% of 4 h LC50 of phyto cercaricides citral, ferulic acid, umbelliferone, azadirachtin and allicin caused significant inhibition of acetylcholinesterase (AChE) and cytochrome oxidase activity in the treated cercaria larva of F. gigantica. Whereas, activity of both enzymes were not significantly altered in the nervous tissues of vector snail Lymnaea acuminata exposed to same treatments. Maximum reduction in AChE (1.35% of control in month of June) and cytochrome oxidase (3.71% of control in the month of July) activity were noted in the cercaria exposed to 60% of 4 h LC50 of azadirachtin and allicin, respectively.

  16. Altered Protein Expression in the Ileum of Mice Associated with the Development of Chronic Infections with Echinostoma caproni (Trematoda)

    PubMed Central

    Cortés, Alba; Sotillo, Javier; Muñoz-Antoli, Carla; Fried, Bernard; Esteban, J. Guillermo; Toledo, Rafael

    2015-01-01

    Background Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode that has been extensively used as experimental model to investigate the factors determining the expulsion of intestinal helminths or, in contrast, the development of chronic infections. Herein, we analyze the changes in protein expression induced by E. caproni infection in ICR mice, a host of high compatibility in which the parasites develop chronic infections. Methodology/Principal Findings To determine the changes in protein expression, a two-dimensional DIGE approach using protein extracts from the intestine of naïve and infected mice was employed; and spots showing significant differential expression were analyzed by mass spectrometry. A total of 37 spots were identified differentially expressed in infected mice (10 were found to be over-expressed and 27 down-regulated). These proteins were related to the restoration of the intestinal epithelium and the control of homeostatic dysregulation, concomitantly with mitochondrial and cytoskeletal proteins among others. Conclusion/Significance Our results suggests that changes in these processes in the ileal epithelium of ICR mice may facilitate the establishment of the parasite and the development of chronic infections. These results may serve to explain the factors determining the development of chronicity in intestinal helminth infection. PMID:26390031

  17. Growth, development and pairing of Leucochloridiomorpha constantiae (Trematoda) metacercariae on the chorio-allantois of chick embryos cultivated in vitro.

    PubMed

    Fried, B; Fine, R H; Felter, B L

    1980-08-01

    A simple in vitro technique was devised to culture chick embryos in Petri dishes from the 4th to the 21st day of incubation. Leucochloridiomorpha constantiae (Trematoda) metacercariae were placed either singly or multiply (5/embryo) on the chorio-allantois of in vitro grown embryos on day 7 and were removed on day 14. Growth and development studies were also made on worms grown singly or multiply (5/chick) in the bursa of Fabricius of the domestic chick. Worms grown singly or multiply in embryos were sexually mature, although eggs from these worms were abnormal when compared with eggs from worms recovered from chicks. The mean body area of worms from chicks was 2-3 times greater than that of worms from embryos. The mean body area of single worms from embryos was significantly larger than that of worms grown multiply in this site. However, the mean body area of multiple worms from the chick was significantly larger than that of single worms from this site. Worm pairs or clusters were seen in all embryos with the multiple infections. PMID:7422365

  18. Development of Mitochondrial Loop-Mediated Isothermal Amplification for Detection of the Small Liver Fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes)

    PubMed Central

    Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van

    2012-01-01

    Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10−4 ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis. PMID:22322346

  19. Development of mitochondrial loop-mediated isothermal amplification for detection of the small liver fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes).

    PubMed

    Le, Thanh Hoa; Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van

    2012-04-01

    Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10(-4) ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis. PMID:22322346

  20. Development of mitochondrial loop-mediated isothermal amplification for detection of the small liver fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes).

    PubMed

    Le, Thanh Hoa; Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van

    2012-04-01

    Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10(-4) ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis.

  1. Comparative assay of glutathione S-transferase (GSTs) activity of excretory-secretory materials and somatic extract of Fasciola spp parasites.

    PubMed

    Alirahmi, Heshmatollah; Farahnak, Ali; Golmohamadi, Taghi; Esharghian, Mohammad Reza

    2010-01-01

    Fascioliasis is a worldwide parasitic disease in human and domestic animals. The causative agents of fascioliasis are Fasciola hepatica and Fasciola gigantica. In the recent years, fasciola resistance to drugs has been reported in the many of publications. Fasciola spp has detoxification system including GST enzyme which may be responsible for its resistance. Therefore , the aim of the study was to assay of GST enzyme activity in fasciola parasites. Fasciola gigantica and Fasciola hepatica helminths were collected from abattoir as a live and cultured in buffer media for 4 h at 37 °C. Excretory-Secretory products were collected and stored in -80◦C. F. gigantica and Fasciola hepatica were homogenized with homogenizing buffer in a glass homogenizer to prepare of somatic extract. Suspension was then centrifuged and supernatant was stored at -80°C. In order to assay the enzyme activity, excretory-secretory and somatic extracts in the form of cocktails (potassium phosphate buffer, reduced glutathione and 1-chloro-2,4-dinitrobenzene substrates) were prepared and their absorbance recorded for 5 minutes at 340 nm. The total and specific GST activity of F. gigantica somatic and ES products were obtained as 2916.00, 272.01 micromole/minute and 1.33, 1.70 micromole/minute/mg protein, respectively. Fasciola hepatica also showed 2705.00, 276.86 micromole/minute and 1.33, 1.52 micromole/minute/mg protein, respectively. These results are important for analysis of parasite survival / resistance to drugs which use for treatment of fascioliasis.

  2. Snail odour-clouds: spreading and contribution to the transmission success of Trichobilharzia ocellata (Trematoda, Digenea) miracidia.

    PubMed

    Hertel, Jan; Holweg, Alexander; Haberl, Bernhard; Kalbe, Martin; Haas, Wilfried

    2006-02-01

    Chemical communication among freshwater organisms is an adaptation to improve their coexistence. Here,we focus on the chemical cues secreted by the freshwater gastropod Lymnaea stagnalis, which are known to stimulate behavioural responses of Trichobilharzia ocellata (Plathelminthes, Digenea, Trematoda) miracidia. Such responses are commonly claimed to influence transmission positively, but in response to chemical cues miracidia randomly change their swimming direction. This kind of response does not necessarily increase transmission, because miracidia may be trapped at the periphery of very large snail odour-clouds, which may prevent them from approaching the snail. On the other hand, the odour clouds may be too small to improve host-localisation. To shed light on these scenarios, the spreading of molecules released around L. stagnalis (active space) was visualised by recording host-finding responses of T. ocellata miracidia when they approached snails. Behavioural responses of miracidia indicated the spreading of compounds forming an attractive active space only around the host-snail L. stagnalis, but not around sympatric non-host-snail species. The active space increased approximately linearly with the time the snail rested at the same spot and within 5 min it reached a volume of more than 30 times that of the snail. We also demonstrated in a large-scale experiment, that the active space of L. stagnalis significantly increases the transmission success of T. ocellata miracidia. Additionally, the microhabitat selection of T. ocellata miracidia was studied, demonstrating that peripheral locations near the water surface were preferred, which are also preferred sites of L. stagnalis. Improved chemoperception and microhabitat selection may have been a consequence of coevolution with snails and benefited miracidia, which became efficient transmissive stages.

  3. Identification of Chalcones as Fasciola hepatica Cathepsin L Inhibitors Using a Comprehensive Experimental and Computational Approach

    PubMed Central

    Ferraro, Florencia; Merlino, Alicia; dell´Oca, Nicolás; Gil, Jorge; Tort, José F.; Gonzalez, Mercedes; Cerecetto, Hugo; Cabrera, Mauricio

    2016-01-01

    Background Increased reports of human infections have led fasciolosis, a widespread disease of cattle and sheep caused by the liver flukes Fasciola hepatica and Fasciola gigantica, to be considered an emerging zoonotic disease. Chemotherapy is the main control measure available, and triclabendazole is the preferred drug since is effective against both juvenile and mature parasites. However, resistance to triclabendazole has been reported in several countries urging the search of new chemical entities and target molecules to control fluke infections. Methodology/Principle Findings We searched a library of forty flavonoid derivatives for inhibitors of key stage specific Fasciola hepatica cysteine proteases (FhCL3 and FhCL1). Chalcones substituted with phenyl and naphtyl groups emerged as good cathepsin L inhibitors, interacting more frequently with two putative binding sites within the active site cleft of the enzymes. One of the compounds, C34, tightly bounds to juvenile specific FhCL3 with an IC50 of 5.6 μM. We demonstrated that C34 is a slow-reversible inhibitor that interacts with the Cys-His catalytic dyad and key S2 and S3 pocket residues, determinants of the substrate specificity of this family of cysteine proteases. Interestingly, C34 induces a reduction in NEJ ability to migrate through the gut wall and a loss of motility phenotype that leads to NEJ death within a week in vitro, while it is not cytotoxic to bovine cells. Conclusions/Significance Up to date there are no reports of in vitro screening for non-peptidic inhibitors of Fasciola hepatica cathepsins, while in general these are considered as the best strategy for in vivo inhibition. We have identified chalcones as novel inhibitors of the two main Cathepsins secreted by juvenile and adult liver flukes. Interestingly, one compound (C34) is highly active towards the juvenile enzyme reducing larval ability to penetrate the gut wall and decreasing NEJ´s viability in vitro. These findings open new avenues

  4. Development of host resistance to Fasciola hepatica after the elimination of primary infection with diamphenethide.

    PubMed

    Corba, J; Spaldonová, R

    1975-01-01

    We studied the specificity of the individual developmental stages of Fasciola hepatica for evoking immune response of the host to reinfection with this parasite, whereby the primary infection was eliminated by a dose of 150 mg/kg diamphenethide administered in various intervals. In rats we observed a state of hypersensitivity demonstrated by retarded migration and growth of the flukes and the reduction in the number of sexually developed parasites. The changes were most marked, if the elimination of the immunizing infection followed 8-10 weeks after primary infection.

  5. Comparative study of sheep and buffalo isolates of Fasciola gigantica in the intermediate host Lymnaea auricularia.

    PubMed

    al-Kubaisee, R Y; Altaif, K I

    1989-09-01

    A comparison was made of the biological behaviour of two isolates of Fasciola gigantica, obtained from sheep and buffalo, in the aquatic snail Lymnaea auricularia. The sheep isolate exhibited a lower rate of infection, slower larval development and the production of fewer cercariae. Larger eggs, miracidia and metacercariae were obtained from the buffalo isolate while the rediae and cercariae were smaller. Metacercariae of sheep origin were dark brown with a mortality after 80 days of storage at 4 degrees C of 30.6 per cent while those of the buffalo isolate were straw coloured and had a mortality of 10.2 per cent.

  6. Characterization of Fasciola spp. in Myanmar on the basis of spermatogenesis status and nuclear and mitochondrial DNA markers.

    PubMed

    Ichikawa, Madoka; Bawn, Saw; Maw, Ni Ni; Htun, Lat Lat; Thein, Myint; Gyi, Aung; Sunn, Kyaw; Katakura, Ken; Itagaki, Tadashi

    2011-12-01

    Fasciola spp. in Myanmar were characterized on the basis of spermatogenesis status and DNA markers of nuclear internal transcribed spacer 1 (ITS1) and mitochondrial NADH dehydrogenase subunit 1 (nad1). We collected 88 adult flukes from Yangon, Lashio, and Myitkyina. Spermatogenesis status was analyzed by the presence of sperm in the seminal vesicles, and 8 aspermic and 80 spermic flukes were detected. The flukes were identified on the basis of spermatogenesis status and ITS1 types which were analyzed by a PCR-RFLP method, and 80 spermic flukes were identified as F. gigantica. A very low detection rate of aspermic Fasciola sp. indicated that they are not established in Myanmar. In phylogenetic analyses, the 7 aspermic Fasciola sp. from Myitkyina displayed a haplotype in nad1 sequence, which was identical to that of aspermic Fasciola sp. from other Asian countries including China. Therefore, they were probably introduced from China through an infected domestic ruminant. On the other hand, 17 nad1 haplotypes detected in F. gigantica belonged to 2 clades unique to Myanmar, each with a distinct founder haplotype in a network analysis. This indicated a unique history of F. gigantica introduction into Myanmar involving ancient artificial movements of domestic ruminants.

  7. LYMPHO-PROLIFERATIVE RESPONSES TO VARIOUS FASCIOLA HEPATICA WORM'S ANTIGENS: AN IN VITRO STUDY.

    PubMed

    Sharaf, Osama F; Amir, Elamir M; Hawash, Yousry A

    2016-04-01

    Fascioliasis is an important zoonotic disease with approximately 2-4 million people infected worldwide and a further 180 million at risk of infection. F. hepatica can survive within the bile ducts for many years through its ability to suppress the host immunity with Fasciola cathepsin L1 cysteine protease and Glutathione S transferase playing an important role. The aim of the present study is to investigate the in vitro lympho-proliferative responses of hepatic hilar lymphocytes (HLN) of infected sheep in response to different F. hepatica antigens. The suppressive effects of Fasciola excretory/secretory (ES) and tegument (TEG) and their fractions were also investigated. Our results showed that both ES and TEG had significant suppressive effects on lympho-proliferation, up to 74% and 92%, respectively. When these antigens were fractionated, fraction 3 (MW of >10000-30000) of both ES (64%) and TEG (59%) in addition to fraction 4 (MW of ≤ 10000) of TEG (38%) inherited the suppressive effects. Identification of the potential molecule(s) with such suppressive effects on lymphocytes in TEG fraction 4 could reveal vaccine candidates. PMID:27363058

  8. Efficacy of temperature, and two commonly used molluscicides and fertilizers on Fasciola gigantica eggs.

    PubMed

    Hassan, Aly A; Shoukary, Nahla M; Ismail, Nahed M M

    2008-08-01

    This study evaluated the effect of temperature, molluscicides (Copper sulphate and Niclosamide), fertilizers (Superphosphate and Ammonium sulphate) on the hatchability of Fasciola gigantica eggs. The results showed that hatchability decreased to 4% when eggs were incubated in bile secretion at 37 degrees C for 5 days and to 1.4% for 10 days, but few eggs incubated in water at 37 degrees C hatched. Bile secretion at 37 degrees C was a poor medium for in-vitro egg preservation. But, hatching occurred only when eggs were transferred to water at 26 degrees C. Temperature fluctuation from 26-4 degrees C or from 32-4 degrees C had an inhibitory effect on embryos development (35.2% & 32.3%, respectively) as compared to controls (60% & 63.9%, respectively). The incubation period (19 & 17 days) was higher than controls (14 & 12 days, respectively). The LC50 & LC90 of Copper sulphate and Niclosamide against Biomphalaria alexandrina and Lymnaea natalenesis had no toxic effect on Fasciola eggs. The higher concentrations of Copper sulphate (30 ppm) and Niclosamide (1 ppm) slightly lower eggs hatchability rate than controls. The rate decreased by increasing the exposure time from 3 to 6 hours with both molluscicides. Ammonium sulphate had a lethal effect on eggs, but Superphosphate had some inhibitory effect on egg development, which increased by increasing Superphosphate concentration or with the prolongation of the exposure time. PMID:18853633

  9. Efficient inhibition of cathepsin B by a secreted type 1 cystatin of Fasciola gigantica.

    PubMed

    Siricoon, Sinee; Grams, Suksiri Vichasri; Grams, Rudi

    2012-12-01

    Cysteine proteases are important antigens in the liver fluke genus Fasciola, essential for infection, protection and nutrition. While their biochemistry, biological roles and application as vaccines have been thoroughly studied there is a lack of data concerning their regulation. In the present study we have continued our investigation of cysteine protease inhibitors in Fasciola gigantica and demonstrate, in comparison with FgStefin-1 and human cystatin C, that a second type 1 cystatin of the parasite, FgStefin-2, has been evolutionary adapted to block cathepsin B. The protein, which unusually for a type 1 cystatin carries a signal peptide, is expressed from the metacercarial to adult stage and located in the epithelial cells of the intestinal tract in all stages and in the prostate gland cells in adults. Both cell types may contribute to the released FgStefin-2 observed in the ES product of the parasite. Distinct isoforms of cathepsin B are essential for host tissue penetration during the early infection process and FgStefin-2 may act as key regulator, required to protect the minute juvenile from autoproteolysis. Expression in the prostate gland in the adult stage suggests an additional regulative role of cysteine protease activity in the reproductive system.

  10. A case of Fasciola hepatica infection mimicking cholangiocarcinoma and ITS-1 sequencing of the worm.

    PubMed

    Kang, Bong Kyun; Jung, Bong-Kwang; Lee, Yoon Suk; Hwang, In Kyeom; Lim, Hyemi; Cho, Jaeeun; Hwang, Jin-Hyeok; Chai, Jong-Yil

    2014-04-01

    Fascioliasis is a zoonotic infection caused by Fasciola hepatica or Fasciola gigantica. We report an 87-year-old Korean male patient with postprandial abdominal pain and discomfort due to F. hepatica infection who was diagnosed and managed by endoscopic retrograde cholangiopancreatography (ERCP) with extraction of 2 worms. At his first visit to the hospital, a gallbladder stone was suspected. CT and magnetic retrograde cholangiopancreatography (MRCP) showed an intraductal mass in the common bile duct (CBD) without proximal duct dilatation. Based on radiological findings, the presumed diagnosis was intraductal cholangiocarcinoma. However, in ERCP which was performed for biliary decompression and tissue diagnosis, movable materials were detected in the CBD. Using a basket, 2 living leaf-like parasites were removed. The worms were morphologically compatible with F. hepatica. To rule out the possibility of the worms to be another morphologically close species, in particular F. gigantica, 1 specimen was processed for genetic analysis of its ITS-1 region. The results showed that the present worms were genetically identical (100%) with F. hepatica but different from F. gigantica.

  11. Phenotypes of intermediate forms of Fasciola hepatica and F. gigantica in buffaloes from Central Punjab, Pakistan.

    PubMed

    Afshan, K; Valero, M A; Qayyum, M; Peixoto, R V; Magraner, A; Mas-Coma, S

    2014-12-01

    Fascioliasis is an important food-borne parasitic disease caused by the two trematode species, Fasciola hepatica and Fasciola gigantica. The phenotypic features of fasciolid adults and eggs infecting buffaloes inhabiting the Central Punjab area, Pakistan, have been studied to characterize fasciolid populations involved. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardized measurements. Since it is the first study of this kind undertaken in Pakistan, the results are compared to pure fasciolid populations: (a) F. hepatica from the European Mediterranean area; and (b) F. gigantica from Burkina Faso; i.e. geographical areas where both species do not co-exist. Only parasites obtained from bovines were used. The multivariate analysis showed that the characteristics, including egg morphometrics, of fasciolids from Central Punjab, Pakistan, are between F. hepatica and F. gigantica standard populations. Similarly, the morphometric measurements of fasciolid eggs from Central Punjab are also between F. hepatica and F. gigantica standard populations. These results demonstrate the existence of fasciolid intermediate forms in endemic areas in Pakistan.

  12. LYMPHO-PROLIFERATIVE RESPONSES TO VARIOUS FASCIOLA HEPATICA WORM'S ANTIGENS: AN IN VITRO STUDY.

    PubMed

    Sharaf, Osama F; Amir, Elamir M; Hawash, Yousry A

    2016-04-01

    Fascioliasis is an important zoonotic disease with approximately 2-4 million people infected worldwide and a further 180 million at risk of infection. F. hepatica can survive within the bile ducts for many years through its ability to suppress the host immunity with Fasciola cathepsin L1 cysteine protease and Glutathione S transferase playing an important role. The aim of the present study is to investigate the in vitro lympho-proliferative responses of hepatic hilar lymphocytes (HLN) of infected sheep in response to different F. hepatica antigens. The suppressive effects of Fasciola excretory/secretory (ES) and tegument (TEG) and their fractions were also investigated. Our results showed that both ES and TEG had significant suppressive effects on lympho-proliferation, up to 74% and 92%, respectively. When these antigens were fractionated, fraction 3 (MW of >10000-30000) of both ES (64%) and TEG (59%) in addition to fraction 4 (MW of ≤ 10000) of TEG (38%) inherited the suppressive effects. Identification of the potential molecule(s) with such suppressive effects on lymphocytes in TEG fraction 4 could reveal vaccine candidates.

  13. Resistance of Fasciola hepatica against triclabendazole in cattle in Patagonia (Argentina).

    PubMed

    Olaechea, F; Lovera, V; Larroza, M; Raffo, F; Cabrera, R

    2011-06-10

    In the winter of 2008, cattle on a farm in the province of Neuquen, Argentina died from subacute and chronic liver fluke disease despite four previous treatments with Triclabendazole (TCBZ). In the spring of 2009, a preliminary efficacy test revealed good performance using nitroxynil, whereas TCBZ efficacy was only 18% by egg counts of Fasciola eggs in the faeces. Resistance to Fasciola hepatica to TCBZ has never been reported in South America, so in January of 2010 a controlled trial was conducted to confirm and to define the degree of resistance in this herd. In a clinical trial, the fluke egg output was monitored on Days 14 and 21 and serum enzymes gamma-glutamyl transpeptidase (GGT) and glutamic-oxaloacetic transaminase (GOT) on Days 0 and 21 in 36 calves treated with TCBZ or with closantel. The results showed a reduction of 100% in fluke egg output at Days 14 and 21 for closantel. The mean epg in the TCBZ-treated groups did not decrease. Because of the fact that in this study TCBZ treatment in cattle had no effect, even at double the recommended dose, it is highly indicative that resistance of F. hepatica against TCBZ is present on this farm. The GGT and GOT levels decreased in the closantel-treated group as a result of the treatment at 21 days after dosing. To evaluate the importance of TCBZ resistance in F. hepatica in Argentina, a study on more farms from endemic areas is needed.

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

    PubMed

    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. PMID:17405628

  15. Phenotypes of intermediate forms of Fasciola hepatica and F. gigantica in buffaloes from Central Punjab, Pakistan.

    PubMed

    Afshan, K; Valero, M A; Qayyum, M; Peixoto, R V; Magraner, A; Mas-Coma, S

    2014-12-01

    Fascioliasis is an important food-borne parasitic disease caused by the two trematode species, Fasciola hepatica and Fasciola gigantica. The phenotypic features of fasciolid adults and eggs infecting buffaloes inhabiting the Central Punjab area, Pakistan, have been studied to characterize fasciolid populations involved. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardized measurements. Since it is the first study of this kind undertaken in Pakistan, the results are compared to pure fasciolid populations: (a) F. hepatica from the European Mediterranean area; and (b) F. gigantica from Burkina Faso; i.e. geographical areas where both species do not co-exist. Only parasites obtained from bovines were used. The multivariate analysis showed that the characteristics, including egg morphometrics, of fasciolids from Central Punjab, Pakistan, are between F. hepatica and F. gigantica standard populations. Similarly, the morphometric measurements of fasciolid eggs from Central Punjab are also between F. hepatica and F. gigantica standard populations. These results demonstrate the existence of fasciolid intermediate forms in endemic areas in Pakistan. PMID:23731975

  16. Evidence of Fasciola spp. resistance to albendazole, triclabendazole and bromofenofos in water buffaloes (Bubalus bubalis).

    PubMed

    Venturina, Virginia M; Alejandro, Ma Antonette F; Baltazar, Cyril P; Abes, Nancy S; Mingala, Claro N

    2015-01-01

    Fasciolosis caused by Fasciola spp. is considered the most important helminth infection of ruminants in tropical countries. Anthelmintic resistance has become a global concern. This study compared the efficacy of the commonly used anthelmintics, determined the toxicity level and any indication of resistance. Thirty two water buffaloes naturally-infected with Fasciola spp. were used to determine the efficacy of triclabendazole (TBZ), albendazole (ABZ), and bromofenofos (BRO) using Fecal Egg Count Reduction Test (FECRT). To test the toxicity of the drugs given, serum glutamic-pyruvic transaminase (SGPT) was evaluated before and within one week after treatment. One dose administration of ABZ registered an efficacy of 79.17%, 73.33% for TBZ and 70.83% for BRO. Efficacy in two dose- treatment group was 83.33% for both BRO and ABZ, and 90.00% for TBZ. Two dose-treatment was effective for TBZ (90%), ineffective for BRO and ABZ. SGPT levels were not significantly different between pre-treatment and post- treatment across all treatments. Giving one or two doses of anthelmintics, at one month interval, does not increase the efficacy of the three drugs tested. The study also implies that anthelmintic resistance may have developed in the animals.

  17. Evidence of Fasciola spp. resistance to albendazole, triclabendazole and bromofenofos in water buffaloes (Bubalus bubalis).

    PubMed

    Venturina, Virginia M; Alejandro, Ma Antonette F; Baltazar, Cyril P; Abes, Nancy S; Mingala, Claro N

    2015-01-01

    Fasciolosis caused by Fasciola spp. is considered the most important helminth infection of ruminants in tropical countries. Anthelmintic resistance has become a global concern. This study compared the efficacy of the commonly used anthelmintics, determined the toxicity level and any indication of resistance. Thirty two water buffaloes naturally-infected with Fasciola spp. were used to determine the efficacy of triclabendazole (TBZ), albendazole (ABZ), and bromofenofos (BRO) using Fecal Egg Count Reduction Test (FECRT). To test the toxicity of the drugs given, serum glutamic-pyruvic transaminase (SGPT) was evaluated before and within one week after treatment. One dose administration of ABZ registered an efficacy of 79.17%, 73.33% for TBZ and 70.83% for BRO. Efficacy in two dose- treatment group was 83.33% for both BRO and ABZ, and 90.00% for TBZ. Two dose-treatment was effective for TBZ (90%), ineffective for BRO and ABZ. SGPT levels were not significantly different between pre-treatment and post- treatment across all treatments. Giving one or two doses of anthelmintics, at one month interval, does not increase the efficacy of the three drugs tested. The study also implies that anthelmintic resistance may have developed in the animals. PMID:26878627

  18. Genotyping and Phylogenetic Analysis of Fasciola Spp. Isolated from Sheep and Cattle Using PCR-RFLP in Ardabil Province, Northwestern Iran

    PubMed Central

    ARYAEIPOUR, Mojgan; ROUHANI, Soheila; BANDEHPOUR, Mojgan; MIRAHMADI, Hadi; KAZEMI, Bahram; ROKNI, Mohammad Bagher

    2014-01-01

    Abstract Background The aim of this study was to detect the genotype of Fasciola spp. in Meshkin-Shahr, Ardabil Province, northwestern Iran in different hosts using PCR-RFLP. Methods The parasite hosts included cattle, and sheep. Overall, 70 adult flukes from livers of slaughtered animals were collected from the abattoirs of aforementioned area. The included 35 samples from infected sheep and 35 samples from 35 infected cattle. PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS 1) region from Fasciola species were used to conduct the study. Results The fragment of approximately 700bp in all of the Fasciola samples was amplified. PCR products of ITS 1 were subjected for digestion by restriction enzyme. RsaI restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. Amplicons with the sequences of F. hepatica had a pattern of about 360, 100, and 60 bp band size, whereas F. gigantica worms had a profile of 360, 170, and 60 bp in size, respectively. Results based on PCR-RFLP analysis were confirmed by sequence analysis of representative ITS 1 amplicons. No hybrid forms were detected in the present study. All sheep were infected with F. hepatica but cattle were infected with both species. Conclusion Both species of Fasciola are present in Ardabil. The method described here can be valuable for identification of Fasciola species in endemic parts for fasciolosis, regions with intermediate species and in that overlapping distribution area. PMID:26060698

  19. RNAi Dynamics in Juvenile Fasciola spp. Liver Flukes Reveals the Persistence of Gene Silencing In Vitro

    PubMed Central

    McVeigh, Paul; McCammick, Erin M.; McCusker, Paul; Morphew, Russell M.; Mousley, Angela; Abidi, Abbas; Saifullah, Khalid M.; Muthusamy, Raman; Gopalakrishnan, Ravikumar; Spithill, Terry W.; Dalton, John P.; Brophy, Peter M.; Marks, Nikki J.; Maule, Aaron G.

    2014-01-01

    Background Fasciola spp. liver fluke cause pernicious disease in humans and animals. Whilst current control is unsustainable due to anthelmintic resistance, gene silencing (RNA interference, RNAi) has the potential to contribute to functional validation of new therapeutic targets. The susceptibility of juvenile Fasciola hepatica to double stranded (ds)RNA-induced RNAi has been reported. To exploit this we probe RNAi dynamics, penetrance and persistence with the aim of building a robust platform for reverse genetics in liver fluke. We describe development of standardised RNAi protocols for a commercially-available liver fluke strain (the US Pacific North West Wild Strain), validated via robust transcriptional silencing of seven virulence genes, with in-depth experimental optimisation of three: cathepsin L (FheCatL) and B (FheCatB) cysteine proteases, and a σ-class glutathione transferase (FheσGST). Methodology/Principal Findings Robust transcriptional silencing of targets in both F. hepatica and Fasciola gigantica juveniles is achievable following exposure to long (200–320 nt) dsRNAs or 27 nt short interfering (si)RNAs. Although juveniles are highly RNAi-susceptible, they display slower transcript and protein knockdown dynamics than those reported previously. Knockdown was detectable following as little as 4h exposure to trigger (target-dependent) and in all cases silencing persisted for ≥25 days following long dsRNA exposure. Combinatorial silencing of three targets by mixing multiple long dsRNAs was similarly efficient. Despite profound transcriptional suppression, we found a significant time-lag before the occurrence of protein suppression; FheσGST and FheCatL protein suppression were only detectable after 9 and 21 days, respectively. Conclusions/Significance In spite of marked variation in knockdown dynamics, we find that a transient exposure to long dsRNA or siRNA triggers robust RNAi penetrance and persistence in liver fluke NEJs supporting the

  20. [The effect of newer anthelmintics on Fasciola hepatica in experimentally infected rats].

    PubMed

    Corba, J

    1976-09-01

    The reports deals with the results of testing seven new antihelminthics for Fasciola hepatica in the experimentally invaded Wistar rat. The greatest influence on juvenile flukes (2 and 4 weeks of age) was exerted by diamphenetid (Coriban) applied in a single dose of 100 mg kg-1. Hexachlorophene applied in the dose of 50 mg kg-1 showed the highest effect on sexually mature flukes. All the tested antihelminthics of the halogenated salicylanilide group were ineffective on juvenile stages and only slightly effective on mature F. hepatica flukes. It follows from the results that the effectiveness of some antifasciolics on laboratory animals need not always be in correlation with their effect in ruminants - hence it is necessary to verify the results obtained in laboratory animals and to check them on natural F. hepatica hosts.

  1. Increase of carboxylesterase activity in Fasciola hepatica recovered from triclabendazole treated sheep.

    PubMed

    Scarcella, S; Miranda-Miranda, E; Cossío-Bayúgar, R; Ceballos, L; Fernandez, V; Solana, H

    2012-10-01

    In the present work, we evaluate in vivo the activity of carboxylesterase of Fasciola hepatica exposed to triclabendazole. We observed a statistically significant increase in enzyme activity at 24 and 48 h post treatment (P<0.01 and P<0.001, respectively). The zymogram of cytosolic fractions identified a protein of 170 kDa containing the carboxylesterase activity. The densitograms of the zymograms confirmed the phenomenon of enzyme induction under the experimental conditions of the assay. These results provide not only the understanding of the importance of this metabolic pathway in flukes but carboxylesterase would also be an enzyme that could participate more actively in the development of anthelmintic resistance at TCBZ.

  2. Epidemiological studies on Fasciola hepatica in Gafsa Oases (south west of Tunisia).

    PubMed

    Hammami, H; Hamed, N; Ayadi, A

    2007-09-01

    Epidemiological investigations on Fasciola hepatica fasciolasis were carried out from July 2004 to June 2005 in the Gafsa oases (Tunisia) after the detection of a human case. Three habitats were studied: one in El Gsar and two in Ain Soltan. The prevalence of human infection was 6.6%. The presence of the parasite was detected through serology in 14.3% of cattle, 35% of sheep and 68.4% of goats. The plants Apium nodiflorum, Oxalis cernua and Sonchus maritimus were suspected to be at the origin of animal contamination and Apium nodiflorum was incriminated in human infection. The prevalence of the infection of the intermediate host Galba truncatula (G. truncatula) was 19.2% from July 2004 to June 2005. Gafsa oases constitute a new location for the development of fasciolasis in the southern west of Tunisia.

  3. Immunomodulatory molecules of Fasciola hepatica: candidates for both vaccine and immunotherapeutic development.

    PubMed

    Dalton, John P; Robinson, Mark W; Mulcahy, Grace; O'Neill, Sandra M; Donnelly, Sheila

    2013-08-01

    The liver fluke, Fasciola hepatica, causes fascioliasis in domestic animals (sheep, cattle), a global disease that is also an important infection of humans. As soon as the parasite invades the gut wall its interaction with various host immune cells (e.g. dendritic cells, macrophages and mast cells) is complex. The parasite secretes a myriad of molecules that direct the immune response towards a favourable non-protective Th2-mediate/regulatory environment. These immunomodulatory molecules, such as cathepsin L peptidase (FhCL1), are under development as the first generation of fluke vaccines. However, this peptidase and other molecules, such as peroxiredoxin (FhPrx) and helminth defence molecule (FhHDM-1), exhibit various immunomodulatory properties that could be harnessed to help treat immune-related conditions in humans and animals. PMID:23623183

  4. Molecular detection of Fasciola hepatica in water sources of District Nowshehra Khyber Pakhtunkhwa Pakistan

    NASA Astrophysics Data System (ADS)

    Khan, Imran; Khan, Amir Muhammad; Ayaz, Khan, Sanaullah; Anees, Muhammad; Khan, Shaukat Ali

    2012-12-01

    Fascioliasis is spread through contamination of water sources and cause morbidity throughout the world. In the current study 300 water samples were processed by PCR for detection of Fasciola hepatica. The overall prevalence in different water sources was 9.66 % (29/300). Highest prevalence was recorded in drain water16 % (16/100) followed by tube well water 10% (4/40), open well water 8 % (8/100) and the lowest was recorded in tap water 1.66 %(1/60). The significant difference P < 0.05 was recorded during data analysis. The highest prevalence was recorded in summer. It was concluded from the study that cleaning and filtration should be adopted to avoid the health hazards against water borne zoonotic parasites.

  5. Temporal genetic variation of Fasciola hepatica from sheep in Galicia (NW Spain).

    PubMed

    Vázquez-Prieto, Severo; Vilas, Román; Ubeira, Florencio M; Paniagua, Esperanza

    2015-04-30

    We found low genetic differentiation between two temporal samples of Fasciola hepatica (2006 and 2008) collected from nine sheep of the same flock that shared the same pasture for at least 2 years. However, each sample, represented by four and five infrapopulations respectively, showed strong heterozygote deficits regarding Hardy-Weinberg expectations and a high degree of genetic structure at infrapopulation level. This is an unexpected result since genetic drift should increase temporal variation among years. Our findings are most likely explained by the fact that the parasite can survive many years in the definitive host. Temporal gene flow favored by high longevity probably increases levels of genetic variability of the population but could also contribute to the observed heterozygote deficits within temporal samples and infrapopulations if it favors the Wahlund effect. Despite the homogenizing effect of gene flow, the high genetic divergence observed between infrapopulations is most likely a consequence of strong genetic drift associated to the complexity of the life cycle.

  6. Ectopic Human Fasciola hepatica Infection by an Adult Worm in the Mesocolon.

    PubMed

    Kim, Ah Jin; Choi, Chang Hwan; Choi, Sun Keun; Shin, Yong Woon; Park, Yun-Kyu; Kim, Lucia; Choi, Suk Jin; Han, Jee Young; Kim, Joon Mee; Chu, Young Chae; Park, In Suh

    2015-12-01

    We report here an ectopic case of Fasciola hepatica infection confirmed by recovery of an adult worm in the mesocolon. A 56-year-old female was admitted to our hospital with discomfort and pain in the left lower quadrant of the abdomen. Abdominal CT showed 3 abscesses in the left upper quadrant, mesentery, and pelvic cavity. On surgical exploration, abscess pockets were found in the mesocolon of the sigmoid colon and transverse colon. A leaf-like worm found in the abscess pocket of the mesocolon of the left colon was diagnosed as an adult fluke of F. hepatica. Histologically, numerous eggs of F. hepatica were noted with acute and chronic granulomatous inflammations in the subserosa and pericolic adipose tissues. Conclusively, a rare case of ectopic fascioliasis has been confirmed in this study by the adult worm recovery of F. hepatica in the mesocolon.

  7. Epidemiological studies on Fasciola hepatica in Gafsa Oases (south west of Tunisia).

    PubMed

    Hammami, H; Hamed, N; Ayadi, A

    2007-09-01

    Epidemiological investigations on Fasciola hepatica fasciolasis were carried out from July 2004 to June 2005 in the Gafsa oases (Tunisia) after the detection of a human case. Three habitats were studied: one in El Gsar and two in Ain Soltan. The prevalence of human infection was 6.6%. The presence of the parasite was detected through serology in 14.3% of cattle, 35% of sheep and 68.4% of goats. The plants Apium nodiflorum, Oxalis cernua and Sonchus maritimus were suspected to be at the origin of animal contamination and Apium nodiflorum was incriminated in human infection. The prevalence of the infection of the intermediate host Galba truncatula (G. truncatula) was 19.2% from July 2004 to June 2005. Gafsa oases constitute a new location for the development of fasciolasis in the southern west of Tunisia. PMID:17933307

  8. Tegumental histological effects of Mirazid® and myrrh volatile oil on adult Fasciola gigantica

    PubMed Central

    Massoud, Ahmad Mohamed; Shalaby, Hatem Abdel Mawgoud; El Khateeb, Rabab Mohamed; Mahmoud, Mona Said; Kutkat, Mohamed Abdel Aziz

    2013-01-01

    Objective To evaluated the histological changes within the tegument of adult Fasciola gigantica (F. gigantica) that led to the gross changes that were visible externally. Methods The effects of oleoresin extract of myrrh (Mirazid®), myrrh volatile oil and triclabendazole sulphoxide (reference drug) on the tegumental structure of adult F. gigantica following treatment in vitro had been determined by light microscopy. Results The internal changes in the tegument observed in this study were compatible with surface changes seen in the previous scanning electron microscopy study, using the same drugs. The swelling of tegumental syncytium was a particular feature of their action, but its level was much greater with myrrh volatile oil, in which vacuolization of the tegument and loss of spines were observed. Conclusions The present study demonstrated the fasciocidal properties of Mirazid® oleoresin extract, and it might be possible to reinforce its fasciocidal activity by increasing its content of myrrh volatile oil. PMID:23730566

  9. Detection of antibodies against Fasciola hepatica in cirrhotic patients from Peru.

    PubMed

    Marcos, L A; Bussalleu, A; Terashima, A; Espinoza, J R

    2009-03-01

    The prevalence of Fasciola hepatica infection, in endemic countries, in patients with established cirrhosis is unknown. We hypothesized that, in endemic countries, the presence of fascioliasis may be detected in a serum pool of cirrhotic patients. Forty-four previously stored serum samples of patients with established liver cirrhosis, in the Hospital Nacional Cayetano Heredia in Lima, Peru, were collected from 1998 to 2003 and assessed for hepatitis B, C and fascioliasis antibodies (Fas2 ELISA). Hepatitis B surface antigen (HBsAg) was positive in 8.8% (n = 34), hepatitis B core antibody (anti-HBc) in 32.5% (n = 34), hepatitis C antibodies (anti-HCV) in 9.1% (n = 33), and 9.1% (n = 44) were Fas2 ELISA positive. This disease is an example of an emerging tropical infection which can be present in chronic liver diseases, requiring greater clinician awareness especially in endemic rural areas. Further clinical studies are warranted.

  10. Ectopic Human Fasciola hepatica Infection by an Adult Worm in the Mesocolon

    PubMed Central

    Kim, Ah Jin; Choi, Chang Hwan; Choi, Sun Keun; Shin, Yong Woon; Park, Yun-Kyu; Kim, Lucia; Choi, Suk Jin; Han, Jee Young; Kim, Joon Mee; Chu, Young Chae; Park, In Suh

    2015-01-01

    We report here an ectopic case of Fasciola hepatica infection confirmed by recovery of an adult worm in the mesocolon. A 56-year-old female was admitted to our hospital with discomfort and pain in the left lower quadrant of the abdomen. Abdominal CT showed 3 abscesses in the left upper quadrant, mesentery, and pelvic cavity. On surgical exploration, abscess pockets were found in the mesocolon of the sigmoid colon and transverse colon. A leaf-like worm found in the abscess pocket of the mesocolon of the left colon was diagnosed as an adult fluke of F. hepatica. Histologically, numerous eggs of F. hepatica were noted with acute and chronic granulomatous inflammations in the subserosa and pericolic adipose tissues. Conclusively, a rare case of ectopic fascioliasis has been confirmed in this study by the adult worm recovery of F. hepatica in the mesocolon. PMID:26797440

  11. Immunomodulatory molecules of Fasciola hepatica: candidates for both vaccine and immunotherapeutic development.

    PubMed

    Dalton, John P; Robinson, Mark W; Mulcahy, Grace; O'Neill, Sandra M; Donnelly, Sheila

    2013-08-01

    The liver fluke, Fasciola hepatica, causes fascioliasis in domestic animals (sheep, cattle), a global disease that is also an important infection of humans. As soon as the parasite invades the gut wall its interaction with various host immune cells (e.g. dendritic cells, macrophages and mast cells) is complex. The parasite secretes a myriad of molecules that direct the immune response towards a favourable non-protective Th2-mediate/regulatory environment. These immunomodulatory molecules, such as cathepsin L peptidase (FhCL1), are under development as the first generation of fluke vaccines. However, this peptidase and other molecules, such as peroxiredoxin (FhPrx) and helminth defence molecule (FhHDM-1), exhibit various immunomodulatory properties that could be harnessed to help treat immune-related conditions in humans and animals.

  12. Hepatic fascioliasis due to Fasciola hepatica: a two-case report.

    PubMed

    Aroonroch, Rangsima; Worawichawong, Suchin; Nitiyanant, Prawat; Kanchanapitak, Auchai; Bunyaratvej, Sukhum

    2006-10-01

    Two cases of hepatic fascioliasis due to Fasciola hepatica were retrieved from our surgical-pathology file since the hospital's foundation in 1969 up to 2005. The diagnosis of hepatic fascioliasis was based on detection of one live fluke in a large cystic lesion in the lobectomized liver specimen in one case and of deposited eggs in the large liver specimen obtained from open biopsy in the other Hepatic fascioliasis is rather rare and almost worldwide in distribution including Thailand. The diagnosis should be considered in the patient from endemic areas consisting of the northern, northeastern and upper-central regions of the country, with a history of ingesting fresh water plants or drinking untreated water and having fever right-upper-quadrant pain or intrahepatic cystic lesion(s) together with absolute peripheral blood eosinophilia.

  13. Glomerulonephritis in water buffaloes (Bubalus bubalis) naturally infected by Fasciola hepatica.

    PubMed

    Marques, Sandra Márcia Tietz; Scroferneker, Maria Lúcia; Edelweiss, Maria Isabel Albano

    2004-08-13

    Glomerulonephritis caused by Fasciola hepatica was observed in buffaloes. Renal biopsies of 20 buffaloes, 11 with F. hepatica and 9 uninfected buffaloes (controls), were examined by light microscopy, direct and indirect immunofluorescence, and immunohistochemical analysis. The biopsies of seven (63.6%) infected buffaloes revealed membranoproliferative glomerulonephritis, three biopsies (27.3%) showed mesangioproliferative glomerulonephritis, and one kidney presented normal biopsy specimens. In the control group, seven buffaloes (77.8%) presented normal biopsy specimens, while two (22.2%) revealed glomerulonephritis-one with a membranoproliferative pattern, and the other with a mesangioproliferative pattern-with extensive inflammatory cell infiltrate. Our conclusion is that glomerulopathy is associated with fascioliasis and that buffaloes are suitable as a naturally existing experimental model of renal injury by circulating immune complexes.

  14. The Complete Mitochondrial Genome of Galba pervia (Gastropoda: Mollusca), an Intermediate Host Snail of Fasciola spp

    PubMed Central

    Huang, Wei-Yi; Zhao, Guang-Hui; Wei, Shu-Jun; Song, Hui-Qun; Xu, Min-Jun; Lin, Rui-Qing; Zhou, Dong-Hui; Zhu, Xing-Quan

    2012-01-01

    Complete mitochondrial (mt) genomes and the gene rearrangements are increasingly used as molecular markers for investigating phylogenetic relationships. Contributing to the complete mt genomes of Gastropoda, especially Pulmonata, we determined the mt genome of the freshwater snail Galba pervia, which is an important intermediate host for Fasciola spp. in China. The complete mt genome of G. pervia is 13,768 bp in length. Its genome is circular, and consists of 37 genes, including 13 genes for proteins, 2 genes for rRNA, 22 genes for tRNA. The mt gene order of G. pervia showed novel arrangement (tRNA-His, tRNA-Gly and tRNA-Tyr change positions and directions) when compared with mt genomes of Pulmonata species sequenced to date, indicating divergence among different species within the Pulmonata. A total of 3655 amino acids were deduced to encode 13 protein genes. The most frequently used amino acid is Leu (15.05%), followed by Phe (11.24%), Ser (10.76%) and IIe (8.346%). Phylogenetic analyses using the concatenated amino acid sequences of the 13 protein-coding genes, with three different computational algorithms (maximum parsimony, maximum likelihood and Bayesian analysis), all revealed that the families Lymnaeidae and Planorbidae are closely related two snail families, consistent with previous classifications based on morphological and molecular studies. The complete mt genome sequence of G. pervia showed a novel gene arrangement and it represents the first sequenced high quality mt genome of the family Lymnaeidae. These novel mtDNA data provide additional genetic markers for studying the epidemiology, population genetics and phylogeographics of freshwater snails, as well as for understanding interplay between the intermediate snail hosts and the intra-mollusca stages of Fasciola spp.. PMID:22844544

  15. Rapid Enhanced MM3-COPRO ELISA for Detection of Fasciola Coproantigens

    PubMed Central

    Martínez-Sernández, Victoria; Orbegozo-Medina, Ricardo A.; González-Warleta, Marta; Mezo, Mercedes; Ubeira, Florencio M.

    2016-01-01

    ELISA-based methods of detecting Fasciola cathepsins in feces are powerful techniques for diagnosing infections by F. hepatica and F. gigantica. In the last decade, the in-house MM3-COPRO ELISA and its commercial version BIO K 201 (BIO X Diagnostics, Belgium) have been recognized as useful tools for detecting early infections by such trematodes and for monitoring the efficacy of anthelmintic treatments in human and animal species, as they provide some advantages over classic fecal egg counts. However, the sensitivity of MM3-COPRO ELISA can sometimes be compromised by the high variability in the concentration of cathepsins in fecal samples throughout the biological cycle of Fasciola (mainly in cattle) and by differences in the between-batch performance of peroxidase-labeled anti-mouse IgG polyclonal antibodies. To prevent such problems, we investigated whether the incorporation of a commercial streptavidin-polymerized horseradish peroxidase conjugate, in order to reveal bound biotinylated monoclonal antibody MM3, can improve the sensitivity of the MM3-COPRO ELISA. We observed that inclusion of this reagent shifted the previous detection limit of the assay from 0.6 ng/mL to 150 pg/mL and that the modified test is able to identify infection in cows harboring only one fluke. Moreover, we demonstrated that maximal OD values can be achieved with short incubations (30 min each step) at RT with shaking, rather than standard incubations, which significantly accelerates the diagnostic procedure. Finally, we did not find a significant correlation between coproantigen concentration and parasite burden in cattle, which may be due to the low parasite burden (1–10 adult flukes) of the animals used in the present study. As the usefulness of the classic MM3-COPRO test for detecting animal and human infections has already been demonstrated, it is expected that the improvements reported in this study will add new insights into the diagnosis and control of fasciolosis. PMID:27438470

  16. Rapid Enhanced MM3-COPRO ELISA for Detection of Fasciola Coproantigens.

    PubMed

    Martínez-Sernández, Victoria; Orbegozo-Medina, Ricardo A; González-Warleta, Marta; Mezo, Mercedes; Ubeira, Florencio M

    2016-07-01

    ELISA-based methods of detecting Fasciola cathepsins in feces are powerful techniques for diagnosing infections by F. hepatica and F. gigantica. In the last decade, the in-house MM3-COPRO ELISA and its commercial version BIO K 201 (BIO X Diagnostics, Belgium) have been recognized as useful tools for detecting early infections by such trematodes and for monitoring the efficacy of anthelmintic treatments in human and animal species, as they provide some advantages over classic fecal egg counts. However, the sensitivity of MM3-COPRO ELISA can sometimes be compromised by the high variability in the concentration of cathepsins in fecal samples throughout the biological cycle of Fasciola (mainly in cattle) and by differences in the between-batch performance of peroxidase-labeled anti-mouse IgG polyclonal antibodies. To prevent such problems, we investigated whether the incorporation of a commercial streptavidin-polymerized horseradish peroxidase conjugate, in order to reveal bound biotinylated monoclonal antibody MM3, can improve the sensitivity of the MM3-COPRO ELISA. We observed that inclusion of this reagent shifted the previous detection limit of the assay from 0.6 ng/mL to 150 pg/mL and that the modified test is able to identify infection in cows harboring only one fluke. Moreover, we demonstrated that maximal OD values can be achieved with short incubations (30 min each step) at RT with shaking, rather than standard incubations, which significantly accelerates the diagnostic procedure. Finally, we did not find a significant correlation between coproantigen concentration and parasite burden in cattle, which may be due to the low parasite burden (1-10 adult flukes) of the animals used in the present study. As the usefulness of the classic MM3-COPRO test for detecting animal and human infections has already been demonstrated, it is expected that the improvements reported in this study will add new insights into the diagnosis and control of fasciolosis. PMID:27438470

  17. Molecular characterization of Fasciola gigantica in Delhi, India and its phylogenetic relation to the species from South Asian countries

    PubMed Central

    HAYASHI, Kei; MOHANTA, Uday K.; NEERAJA, Tambireddy; ITAGAKI, Tadashi

    2016-01-01

    The aim of this study was to phylogenetically analyze Fasciola gigantica (F. gigantica) from mainland India and to reveal the expansion history of F. gigantica in the Indian subcontinent. We analyzed 40 Fasciola flukes that were collected from Delhi, in the Indian mainland, and identified them as F. gigantica by using nucleotide analyses of the nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes. Based on the nucleotide sequence of mitochondrial NADH dehydrogenase subunit 1 (nad1) gene, the flukes had 18 haplotypes. The haplotypes were classified under haplogroup A, which is predominant in the F. gigantica of South Asia. The population genetics of haplogroup A revealed that Delhi population showed higher π value than eastern India population. These results suggest that F. gigantica of haplogroup A might have spread from the west to the east in India along with the artificial migration of the domestic Zebu cattle, Bos indicus. PMID:27301703

  18. Fasciola gigantica: cercarial shedding pattern from Lymnaea natalensis after long-term exposure to cadmium at different temperatures.

    PubMed

    Soliman, Maha F M

    2009-04-01

    Little is known about the response of Fasciola spp., inside its snail host to pollutant toxicity. The effect of long-term exposure of Lymnaea natalensis to 0.1 microg/l, 10 microg/l, or 100 microg/l cadmium (Cd) on the infection rates with F. gigantica, percentage of cercariae-shedding snails and cercariae shedding pattern was investigated. The snails were exposed to Cd for 7 days either pre-infection with single Fasciola miracidium or during the late pre-patent period of the infection. The possible interaction between metal exposure and acclimatization temperature was also studied in three ranges; 16-18 degrees C, 23-25 degrees C and 30-32 degrees C. Results clearly showed that host exposure to 100 microg/l Cd significantly affected the pattern of Fasciola development inside its snail host, and that the acclimatization temperature was a key factor affecting the role played by Cd. Pre-infection exposure to Cd caused a significant reduction in the infection rates where the effect was temperature-dependent. Post-infection exposure to Cd significantly increased the percentage of cercariae-shedding snails; the effect was temperature-independent. Interestingly, post-infection exposure significantly altered the differential cercarial output pattern, while no clear impact on the total was found. The alteration in the differential cercarial output was represented as a significantly higher proportion of floating metacercaria as compared to fixed ones and also higher proportion of dead cercariae which may directly reduce the transmission of Fasciola to the final host. The mechanisms of cadmium impact are briefly discussed. PMID:19101545

  19. First insight into CD59-like molecules of adult Fasciola hepatica.

    PubMed

    Shi, Yunliang; Toet, Hayley; Rathinasamy, Vignesh; Young, Neil D; Gasser, Robin B; Beddoe, Travis; Huang, Weiyi; Spithill, Terry W

    2014-09-01

    The present study focussed on investigating CD59-like molecules of Fasciola hepatica. A cDNA encoding a CD59-like protein (termed FhCD59-1) identified previously in the membrane fraction of the F. hepatica tegument was isolated. This homologue was shown to encode a predicted open reading frame (ORF) of 122 amino acids (aa) orthologous to human CD59 with a 25 aa signal peptide, a mature protein containing 10 cysteines and a conserved CD59/Ly-6 family motif "CCXXXXCN". An analysis of cDNAs from two different adult specimens of F. hepatica revealed seven variable types of FhCD59-1 sequences, designated FhCD59-1.1 to FhCD59-1.7, which had 94.3-99.7% amino acid sequence identity upon pairwise comparison. Molecular modeling of FhCD59-1.1 with human CD59 confirmed the presence of the three-finger protein domain found in the CD59 family and predicted three disulphide bonds in the F. hepatica sequence. The interrogation of F. hepatica databases identified two additional sequences, designated FhCD59-2 and FhCD59-3, which had only 23.4-29.5% amino acid identity to FhCD59-1.1. Orthologues of the inferred CD59 protein sequences of F. hepatica were also identified in other flatworms, including Fasciola gigantica, Fascioloides magna, Schistosoma haematobium, Schistosoma japonicum, Schistosoma mansoni, Clonorchis sinensis, Opisthorchis viverrini, Taenia solium, Echinococcus granulosus and the free living Schmidtea mediterannea. The results revealed a considerable degree of sequence complexity in the CD59-like sequence families in F. hepatica and flatworms. Phylogenetic analysis of CD59-like aa sequences from F. hepatica and flatworms showed that FhCD59-2 clustered with the known surface-associated protein SmCD59-2 of S. mansoni. Relatively well-supported clades specific to schistosomes, fasciolids and opisthorchiids were identified. The qPCR analysis of gene transcription showed that the relative expression of these 3 FhCD59-like sequences varied by 11-47-fold during fluke

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

    PubMed Central

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

    2015-01-01

    Background 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. 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). Results 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. Conclusion 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

  1. First molecular identification of Australapatemon burti (Miller, 1923) (Trematoda: Digenea: Strigeidae) from an intermediate host Radix labiata (Rossmaessler) (Gastropoda: Lymnaeidae) in Europe.

    PubMed

    Aksenova, Olga V; Bespalaya, Yulia V; Bolotov, Ivan N; Kondakov, Alexander V; Sokolova, Svetlana E

    2016-01-01

    The strigeid digenean species Australapatemon burti (Miller, 1923) (Trematoda: Digenea: Strigeidae) was originally described from North America, but recorded in the Neotropical region (Drago et al. 2007; Hernández-Mena et al. 2014; Blasco-Costa et al. 2016) and in Central Europe (Faltýnková et al. 2007). In Europe, this species is rare, and there is not much information about its range (Faltýnková et al. 2007; Soldánová et al. 2012). Australapatemon burti has a complex life cycle with three larval stages, two of which (sporocyst and cercaria) use several species of freshwater snails, and the third stage (metacercaria) use non-specific host hirudineans (Dubois 1968; Davies & Ostrowski de Núñez 2012; Blasco-Costa et al. 2016). Adult flukes are parasitic in the intenstines of various waterfowl species, such as ducks and swans (Drago et al. 2007; Hernández-Mena et al. 2014). Currently, the molecular data on this parasite species includes only nucleotide sequences of four adult specimens from Mexico (Hernández-Mena et al. 2014). Their hosts were Mexican duck, Anas diazi Ridgway, American Wigeon, Anas americana Gmelin, Cinnamon Teal, Anas cyanoptera Vieillot, and Ruddy Duck, Oxyura jamaicensis (Gmelin) (Anserformes: Anatidae). PMID:27395696

  2. Fluctuations in densities of the invasive gill parasite Centrocestus formosanus (Trematoda: Heterophyidae) in the Comal River, Comal County, Texas, U.S.A.

    PubMed

    Johnson, Matthew S; Bolick, Anne; Alexander, Mara; Huffman, David; Oborny, Ed; Monroe, Allen

    2012-02-01

    Centrocestus formosanus (Trematoda: Heterophyidae) is an invasive fish parasite in the Comal River, Texas, and is considered a threat to the federally endangered fountain darter, Etheostoma fonticola . Monitoring densities of C. formosanus cercariae is crucial to determining levels of infection pressure. We sampled 3 sites in the Comal River during 2 sampling periods, the first during 2006-2007, and again during 2009-2010. Two of the sites were located in the upstream reach of Landa Lake, sites HS and LA, and the third site was located downstream of Landa Lake in the old channel of the river. Cercariae densities were highest at the downstream most site (EA), followed by sites LA and HS, during both sampling periods, but a significant decline in cercariae density was observed between the first and second sampling periods. Several abiotic factors were monitored, including total stream discharge, wading discharge, temperature, and dissolved oxygen, but no river-wide trends were observed. Therefore, we speculate that these factors do not adequately explain the observed long-term decline in cercariae density. We propose that the decline is simply a reflection of a typical pattern followed by most invasive species as they gradually become integrated into the local community following an initial explosive growth in population size. Although cercariae densities may be abating, fountain darters in the Comal River are still threatened by the parasite, and conservation efforts must focus on reducing levels of infection pressure from the parasite whenever possible.

  3. Local adaptation of the trematode Fasciola hepatica to the snail Galba truncatula

    PubMed Central

    Dreyfuss, G.; Vignoles, P.; Rondelaud, D.

    2012-01-01

    Experimental infections of six riverbank populations of Galba truncatula with Fasciola hepatica were carried out to determine if the poor susceptibility of these populations to this digenean might be due to the scarcity or the absence of natural encounters between these snails and the parasite. The first three populations originated from banks frequented by cattle in the past (riverbank group) whereas the three others were living on islet banks without any known contact with local ruminants (islet group). After their exposure, all snails were placed in their natural habitats from the end of October up to their collection at the beginning of April. Compared to the riverbank group, snails, which died without cercarial shedding clearly predominated in the islet group, while the other infected snails were few in number. Most of these last snails released their cercariae during a single shedding wave. In islet snails dissected after their death, the redial and cercarial burdens were significantly lower than those noted in riverbank G. truncatula. Snails living on these islet banks are thus able to sustain larval development of F. hepatica. The modifications noted in the characteristics of snail infection suggest the existence of an incomplete adaptation between these G. truncatula and the parasite, probably due to the absence of natural contact between host and parasite. PMID:22910670

  4. Occurrence of Fasciola hepatica (Linnaeus, 1758) in capybara (Hydrochoerus hydrochaeris) (Linnaeus, 1766) in Minas Gerais, Brazil.

    PubMed

    Dracz, Ruth Massote; Ribeiro, Vinicius Marques Antunes; Pereira, Cintia Aparecida de Jesus; Lima, Walter Dos Santos

    2016-01-01

    Fasciola hepatica is a parasite that affects the hepatic ducts of several species of domestic and wild vertebrates, causing huge economic losses to livestock rearing worldwide. Reports on occurrences of F. hepatica in capybaras are an important epidemiological aspect of this disease, since these rodents can be a source of contamination for other animals and humans. In the present study, conducted in a rural area of the municipality of Confins, Minas Gerais, fresh feces from capybaras were collected from the ground near a lagoon at the edge of the Ribeirão da Mata river. These were examined using the technique of four metal sieves. F. hepatica eggs were recovered. This trematode species was confirmed by observing morphological characteristics and measuring the eggs recovered from the capybara feces, and through experimental infection of Lymnaea columella (Say, 1817) by miracidia from these eggs and subsequent infection of C57/BL06 mice with metacercariae originating from these infected mollusks. The data suggest the occurrence of natural cycle of F. hepatica in this region and provide a warning that expansion of the geographical distribution of this parasite by means of this rodent is possible. It is therefore important to adopting measures for epidemiological control of this helminthiasis.

  5. An enzyme-linked immunosorbent assay for diagnosis of Fasciola gigantica infection in cattle and buffaloes.

    PubMed

    Krishna Murthy, C M; Souza, Placid E D

    2015-12-01

    The enzyme-linked immunosorbent assay (ELISA) was evaluated for the diagnosis of Fasciola gigantica infection in cattle and buffaloes. The excretory-secretory (E-S Ag) antigen of F. gigantica adult flukes obtained after invitro incubation was used as an antigen. The test was conducted with 276 sera collected from cattle and buffaloes which included 22 sera each from naturally infected cattle and buffaloes (known positive serum) and with similar number of samples with healthy cattle and buffaloes (known negative serum). The positive results were observed in 18 and 19 of the sera from naturally infected cattle and buffaloes with sensitivity of 81.8 and 86.3 % respectively. Out of 188 serum samples which were found negative on faecal examination 32 (34 %) sera of cattle and 40 (42.5 %) sera of buffaloes were found positive by ELISA respectively. The sensitivity of the test was found to be 91.6 and 95.6 % in cattle and buffaloes respectively.

  6. Epidemiology and impact of Fasciola hepatica exposure in high-yielding dairy herds

    PubMed Central

    Howell, Alison; Baylis, Matthew; Smith, Rob; Pinchbeck, Gina; Williams, Diana

    2015-01-01

    The liver fluke Fasciola hepatica is a trematode parasite with a worldwide distribution and is the cause of important production losses in the dairy industry. The aim of this observational study was to assess the prevalence of exposure to F. hepatica in a group of high yielding dairy herds, to determine the risk factors and investigate their associations with production and fertility parameters. Bulk milk tank samples from 606 herds that supply a single retailer with liquid milk were tested with an antibody ELISA for F. hepatica. Multivariable linear regression was used to investigate the effect of farm management and environmental risk factors on F. hepatica exposure. Higher rainfall, grazing boggy pasture, presence of beef cattle on farm, access to a stream or pond and smaller herd size were associated with an increased risk of exposure. Univariable regression was used to look for associations between fluke exposure and production-related variables including milk yield, composition, somatic cell count and calving index. Although causation cannot be assumed, a significant (p < 0.001) negative association was seen between F. hepatica exposure and estimated milk yield at the herd level, representing a 15% decrease in yield for an increase in F. hepatica exposure from the 25th to the 75th percentile. This remained significant when fertility, farm management and environmental factors were controlled for. No associations were found between F. hepatica exposure and any of the other production, disease or fertility variables. PMID:26093971

  7. Differential expression and localization of saposin-like protein 2 of Fasciola hepatica

    PubMed Central

    Cabán-Hernández, Kimberly; Espino, Ana M.

    2013-01-01

    FhSAP2 is a novel antigen isolated from the adult fluke of Fasciola hepatica. Based on sequence similarity with amoebapores and other related proteins, it belongs to the saposin-like protein (SAPLIP) family. FhSAP2 has been shown to be highly immunogenic and capable of inducing protective immune responses in mice and rabbits challenged with F. hepatica. Moreover, FhSAP2 is also reactive with sera from humans with chronic fascioliasis. In the present study, we investigated the expression of FhSAP2 in various developmental stages of F. hepatica by qPCR and demonstrated that FhSAP2-mRNA species are up-regulated in undeveloped eggs, newly excysted juveniles, and adults, but down-regulated in the miracidium stage. Monoclonal antibodies against FhSAP2 were produced, and two clones that are positive to F. hepatica whole-body extract, but not reactive with extracts from other trematodes, were selected, expanded and used for histolocalization studies. Confocal immunofluorescence revealed the presence of native FhSAP2 in epithelial cells surrounding the gut, toward the outermost part of the tegument, and toward the tegumental cells of both adults and newly excysted juveniles. PMID:23988299

  8. Characterization and differential expression of a ferritin protein from Fasciola hepatica

    PubMed Central

    Cabán-Hernández, Kimberly; Gaudier, José F.; Espino, Ana M.

    2012-01-01

    Ferritins are proteins that play a central role in maintaining intracellular iron balance. A cDNA clone of Fasciola hepatica (687 bp long) encoding a putative 228-amino acid polypeptide (FhFtn-1) homologous with ferritins of vertebrates and invertebrates was identified. FhFtn-1 contains a conserved motif of the ferroxidase center typical of vertebrate ferritins. Phylogenetic tree analysis showed that FhFtn-1 clusters with two ferritins of Paragonimus westermani, which suggests a common ancestry for the ferritins of these two trematodes. Recombinant FhFtn-1 protein expressed and purified from an Escherichia coli system showed iron-uptake ability. Moreover, FhFtn-1 showed strong reactivity with sera from rabbits infected with F. hepatica for 2–12 weeks, which suggests that this protein could be a potential antigen for immunodiagnosis of fascioliasis. qPCR analysis demonstrated that FhFtn-1-mRNA is expressed at significantly higher levels in adults and unembryonated eggs than in juveniles or miracidia. These results represent the first characterization of a ferritin protein from the liver fluke F. hepatica. PMID:22240114

  9. Activity of artemether and OZ78 against triclabendazole-resistant Fasciola hepatica.

    PubMed

    Keiser, Jennifer; Utzinger, Jürg; Vennerstrom, Jonathan L; Dong, Yuxiang; Brennan, Gerry; Fairweather, Ian

    2007-12-01

    Triclabendazole is the drug of choice against Fasciola hepatica infections in humans and animals. However, parasite resistance against triclabendazole is spreading in the veterinary field, and there are no drugs of comparable activity currently available for the treatment and control of fascioliasis. We investigated the efficacy of single oral doses of artemether and OZ78 against adult triclabendazole-resistant F. hepatica harboured in rats, and compared the results with triclabendazole administered at two different doses. Single oral doses of 100 mg/kg OZ78 and 200 mg/kg artemether resulted in worm burden reductions of 100%. Whereas a single 10 mg/kg dose of triclabendazole achieved a worm burden reduction of only 4.0%, a five-fold higher dose yielded a significant worm burden reduction of 60.9%. However, the lower dose of triclabendazole administered to rats harbouring a triclabendazole-sensitive F. hepatica isolate resulted in a worm burden reduction of 95.3%. Our findings confirm that artemether and OZ78 possess good fasciocidal properties, even against a triclabendazole-resistant F. hepatica isolate, and hence these drugs might become useful in areas where triclabendazole resistance is common.

  10. In vivo assessment of closantel ovicidal activity in Fasciola hepatica eggs.

    PubMed

    Solana, María Victoria; Mera y Sierra, Roberto; Scarcella, Silvana; Neira, Gisela; Solana, Hugo Daniel

    2016-01-01

    Anthelmintic resistance in livestock parasites is currently a worldwide problem. Fasciola hepatica is a cosmopolitan parasite which causes considerable loss in sheep and cattle production systems all over the world. Chemotherapy is currently the main tool available for its control. The intensive use of triclabendazole, the drug of choice for more than 20 years, has resulted in the development of resistant strains. The therapeutic options are adulticides such as closantel (salicylanilide anthelmintic that binds extensively to plasma albumin) to treat chronic fascioliasis in sheep, and cattle. In the present work, an Egg Hatch Assay (EHA) and morphometric studies were used to evaluate in vivo the ovicidal activity and morphology F. hepatica eggs, recovered from closantel treated sheep collected at different time intervals post treatment. Statistically significant differences (p < 0.0001) were observed in egg morphometry between the control and the treated groups in all the parameters studied. Eggs recovered from treated animals tend to be narrower and longer. Significant differences were found in the embryonation and hatching of eggs between 36 h post treatment (32, 5%) vs. approximately 85% in control, 12 h and 24 h post treatment. Our results confirm that closantel affects in vivo the normal development of the eggs. As one of the first effects, this drug affects the performance of the trematode's reproductive physiology. Even though closantel treated animals may still eliminate eggs in the first days post treatment, these are not viable. PMID:26551411

  11. In vivo assessment of closantel ovicidal activity in Fasciola hepatica eggs.

    PubMed

    Solana, María Victoria; Mera y Sierra, Roberto; Scarcella, Silvana; Neira, Gisela; Solana, Hugo Daniel

    2016-01-01

    Anthelmintic resistance in livestock parasites is currently a worldwide problem. Fasciola hepatica is a cosmopolitan parasite which causes considerable loss in sheep and cattle production systems all over the world. Chemotherapy is currently the main tool available for its control. The intensive use of triclabendazole, the drug of choice for more than 20 years, has resulted in the development of resistant strains. The therapeutic options are adulticides such as closantel (salicylanilide anthelmintic that binds extensively to plasma albumin) to treat chronic fascioliasis in sheep, and cattle. In the present work, an Egg Hatch Assay (EHA) and morphometric studies were used to evaluate in vivo the ovicidal activity and morphology F. hepatica eggs, recovered from closantel treated sheep collected at different time intervals post treatment. Statistically significant differences (p < 0.0001) were observed in egg morphometry between the control and the treated groups in all the parameters studied. Eggs recovered from treated animals tend to be narrower and longer. Significant differences were found in the embryonation and hatching of eggs between 36 h post treatment (32, 5%) vs. approximately 85% in control, 12 h and 24 h post treatment. Our results confirm that closantel affects in vivo the normal development of the eggs. As one of the first effects, this drug affects the performance of the trematode's reproductive physiology. Even though closantel treated animals may still eliminate eggs in the first days post treatment, these are not viable.

  12. Across intra-mammalian stages of the liver f luke Fasciola hepatica: a proteomic study

    NASA Astrophysics Data System (ADS)

    di Maggio, Lucía Sánchez; Tirloni, Lucas; Pinto, Antonio F. M.; Diedrich, Jolene K.; Yates, John R., III; Benavides, Uruguaysito; Carmona, Carlos; da Silva Vaz, Itabajara, Jr.; Berasain, Patricia

    2016-09-01

    Fasciola hepatica is the agent of fasciolosis, a foodborne zoonosis that affects livestock production and human health. Although flukicidal drugs are available, re-infection and expanding resistance to triclabendazole demand new control strategies. Understanding the molecular mechanisms underlying the complex interaction with the mammalian host could provide relevant clues, aiding the search for novel targets in diagnosis and control of fasciolosis. Parasite survival in the mammalian host is mediated by parasite compounds released during infection, known as excretory/secretory (E/S) products. E/S products are thought to protect parasites from host responses, allowing them to survive for a long period in the vertebrate host. This work provides in-depth proteomic analysis of F. hepatica intra-mammalian stages, and represents the largest number of proteins identified to date for this species. Functional classification revealed the presence of proteins involved in different biological processes, many of which represent original findings for this organism and are important for parasite survival within the host. These results could lead to a better comprehension of host-parasite relationships, and contribute to the development of drugs or vaccines against this parasite.

  13. Fasciola hepatica miracidia: lectin binding and stimulation of in vitro miracidium-to-sporocyst transformation.

    PubMed

    Georgieva, Katya; Georgieva, Simona; Mizinska, Yana; Stoitsova, Stoyanka R

    2012-03-01

    The lectin binding properties of Fasciola hepatica miracidia were studied by a panel of fluorescein- and gold-conjugated lectins (ConA, LCA, WGA, LEA, SBA, HPA and UEA-I). The presence of mannose and/or glucose residues was demonstrated with ConA and LCA as weak diffuse fluorescence of the miracidial surface, which was more intense at the anterior part of the larva. The N-acetylglucosamine-binding lectins WGA and LEA reacted intensely with the whole miracidial surface. No labelling with N-acetylgalactosamine and/or galactose-specific (SBA and HPA) and fucose-specific UEA-I lectins was observed. The possibility that the specific recognition of the miracidial surface carbohydrates by lectins may initiate the process of transformation of the miracidia into sporocysts was examined in vitro in physiological saline for Galba truncatula. Incubation in the presence of ConA and WGA resulted in facilitation of the transformation process. Facilitation was absent in the presence of inhibitor sugars. Incubation in the presence of SBA or UEA-I had no effect. The results suggested a possible impact of carbohydrate-lectin interactions in transformation of miracidia of F. hepatica to sporocysts in vivo. PMID:22807013

  14. Motility of Fasciola hepatica miracidia assessed with a computer-assisted sperm analyser.

    PubMed

    Villa-Mancera, A; Reynoso-Palomar, A; Olivares-Pérez, J; Ortega-Vargas, S; Cruz-Mendoza, I; Quiroz-Romero, H

    2015-07-01

    The motility parameters of Fasciola hepatica miracidia were assessed at different temperatures and times post-hatching using computer-assisted sperm analysis. Eggs were incubated at 22 °C or 25 °C for 14 days. Five motion parameters were evaluated at different incubation temperatures up to 10 h post-hatching. No differences were observed in the percentage that hatched after incubation at the two different temperatures. However, the straight-line velocity of miracidia following incubation at 22 °C was significantly different from that observed at 25 °C (P< 0.01). All miracidium motion parameters at different post-hatching temperatures showed an overall decrease at the end of the experiment. Those miracidia hatching from eggs incubated at 25 °C had a higher velocity of 1673.3 μm/s compared with 1553.3 μm/s at 22 °C. Velocity parameters increased as the post-hatching temperature increased from 22 °C to 37 °C. PMID:26017332

  15. Differential expression and localization of saposin-like protein 2 of Fasciola hepatica.

    PubMed

    Cabán-Hernández, Kimberly; Espino, Ana M

    2013-12-01

    FhSAP2 is a novel antigen isolated from the adult fluke of Fasciola hepatica. Based on sequence similarity with amoebapores and other related proteins, it belongs to the saposin-like protein (SAPLIP) family. FhSAP2 has been shown to be highly immunogenic and capable of inducing protective immune responses in mice and rabbits challenged with F. hepatica. Moreover, FhSAP2 is also reactive with sera from humans with chronic fascioliasis. In the present study, we investigated the expression of FhSAP2 in various developmental stages of F. hepatica by qPCR and demonstrated that FhSAP2-mRNA species are up-regulated in undeveloped eggs, newly excysted juveniles, and adults, but down-regulated in the miracidium stage. Monoclonal antibodies against FhSAP2 were produced, and two clones that are positive to F. hepatica whole-body extract, but not reactive with extracts from other trematodes, were selected, expanded and used for histolocalization studies. Confocal immunofluorescence revealed the presence of native FhSAP2 in epithelial cells surrounding the gut, toward the outermost part of the tegument, and toward the tegumental cells of both adults and newly excysted juveniles. PMID:23988299

  16. Development and hatching mechanism of Fasciola eggs, light and scanning electron microscopic studies.

    PubMed

    Hussein, Abdel-Nasser A; Hassan, Ismael M; Khalifa, Refaat M A

    2010-07-01

    Both light microscopy and scanning electron microscopy were used in the description. There were variable measurements of eggs from the same fluke and there was no relationship between the size of the fluke and size of eggs. Light microscopy revealed that the operculum has different shapes in Fasciola gigantica and F. hepatica. Under normal laboratory conditions of temperature (26 ± 1 °C), miracidia of F. gigantica developed within 12-16 days period, but those of F. hepatica developed within a period of 13-15 days. The miracidia of F. gigantica measured 110 × 70 μm and that of F. hepatica measured 136 × 74 μm. The life span of the miracidium in F. gigantica ranged between 9 and 12 h, while in F. hepatica the life span did not exceed 10 h. Miracidial epidermal plates in miracidium of F. gigantica were found to be 20 plates arranged in four tiers of six, four, six and four. Hatching process was recorded using scanning electron microscope, and it indicated partially opened operculum. PMID:23961086

  17. Fasciola hepatica: development of redial generations in experimental infections of Pseudosuccinea columella.

    PubMed

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

    2014-07-01

    Experimental infections of Egyptian Pseudosuccinea columella with one or two miracidia of Fasciola hepatica per snail were carried out to determine the developmental pattern (normal or abnormal) of redial generations and specify the number of free rediae developing in snails according to their generation. Controls were constituted by a French population of Galba truncatula infected according to the same protocol. Most infected P. columella showed a normal development of redial generations (96.2-98.1 vs 75.5-85.7% for G. truncatula). In each redial category, free rediae were more numerous in P. columella than in G. truncatula, and their number were also greater in the two-miracidia groups than in single-miracidium infections for each lymnaeid considered separately. This increase in redial production was mainly due to the number of first mother (R1a) rediae producing daughter rediae only: 2 per P. columella (vs one redia in G. truncatula) in single-miracidium groups and 3.1 (vs 1.9) in the two-miracidia groups. In P. columella, the mean total number of free rediae developing in single-miracidium and bimiracidial infections was 77.2 and 117.6, respectively (instead of 33.5 and 52.1 rediae in G. truncatula). The number of F. hepatica rediae present in P. columella was related to the number of fully grown sporocysts and the quantity of R1a rediae which developed into the snail body. PMID:24832813

  18. Study of surface carbohydrates in Galba truncatula tissues before and after infection with Fasciola hepatica

    PubMed Central

    Georgieva, Katya; Georgieva, Liliya; Mizinska-Boevska, Yana; Stoitsova, Stoyanka R

    2016-01-01

    The presence and distribution of surface carbohydrates in the tissues of Galba truncatula snails uninfected or after infection with Fasciola hepatica as well as on the surface of the snail-pathogenic larval stages of the parasite were studied by lectin labelling assay. This is an attempt to find similarities that indicate possible mimicry, utilised by the parasite as an evasion strategy in this snail-trematode system. Different binding patterns were identified on head-foot-mantle, hepatopancreas, genital glands, renopericardial complex of the host as well as of the snail-pathogenic larval stages of F. hepatica. The infection with F. hepatica leads to changes of labelling with Glycine max in the head-mantle cells and Arachis hypogaea in the tubular epithelium of the hepatopancreas. The lectin binding on the other snail tissues is not changed by the development of the larvae. Our data clearly demonstrated the similarity in labelling of G. truncatula tissues and the surface of the snail-pathogenic larval stages of F. hepatica. The role of glycosylation of the contact surfaces of both organisms in relation to the host-parasite interactions is also discussed. PMID:27384082

  19. Fasciola hepatica IN BOVINES IN BRAZIL: DATA AVAILABILITY AND SPATIAL DISTRIBUTION

    PubMed Central

    Bennema, Sita C.; Scholte, Ronaldo Guilherme Carvalho; Molento, Marcelo Beltrão; Medeiros, Camilla; Carvalho, Omar dos Santos

    2014-01-01

    Fasciolosis is a disease of importance for both veterinary and public health. For the first time, georeferenced prevalence data of Fasciola hepatica in bovines were collected and mapped for the Brazilian territory and data availability was discussed. Bovine fasciolosis in Brazil is monitored on a Federal, State and Municipal level, and to improve monitoring it is essential to combine the data collected on these three levels into one dataset. Data were collected for 1032 municipalities where livers were condemned by the Federal Inspection Service (MAPA/SIF) because of the presence of F. hepatica. The information was distributed over 11 states: Espírito Santo, Goiás, Minas Gerais, Mato Grosso do Sul, Mato Grosso, Pará, Paraná, Rio de Janeiro, Rio Grande do Sul, Santa Catarina and São Paulo. The highest prevalence of fasciolosis was observed in the southern states, with disease clusters along the coast of Paraná and Santa Catarina and in Rio Grande do Sul. Also, temporal variation of the prevalence was observed. The observed prevalence and the kriged prevalence maps presented in this paper can assist both animal and human health workers in estimating the risk of infection in their state or municipality. PMID:24553606

  20. Study of surface carbohydrates in Galba truncatula tissues before and after infection with Fasciola hepatica.

    PubMed

    Georgieva, Katya; Georgieva, Liliya; Mizinska-Boevska, Yana; Stoitsova, Stoyanka R

    2016-07-01

    The presence and distribution of surface carbohydrates in the tissues of Galba truncatula snails uninfected or after infection with Fasciola hepatica as well as on the surface of the snail-pathogenic larval stages of the parasite were studied by lectin labelling assay. This is an attempt to find similarities that indicate possible mimicry, utilised by the parasite as an evasion strategy in this snail-trematode system. Different binding patterns were identified on head-foot-mantle, hepatopancreas, genital glands, renopericardial complex of the host as well as of the snail-pathogenic larval stages of F. hepatica. The infection with F. hepatica leads to changes of labelling with Glycine max in the head-mantle cells and Arachis hypogaea in the tubular epithelium of the hepatopancreas. The lectin binding on the other snail tissues is not changed by the development of the larvae. Our data clearly demonstrated the similarity in labelling of G. truncatula tissues and the surface of the snail-pathogenic larval stages of F. hepatica. The role of glycosylation of the contact surfaces of both organisms in relation to the host-parasite interactions is also discussed. PMID:27384082

  1. A multiplex PCR for the detection of Fasciola hepatica in the intermediate snail host Galba cubensis.

    PubMed

    Alba, Annia; Vázquez, Antonio A; Hernández, Hilda; Sánchez, Jorge; Marcet, Ricardo; Figueredo, Mabel; Sarracent, Jorge; Fraga, Jorge

    2015-07-30

    Fasciolosis is a snail-borne trematode infection that has re-emerged as a human disease, and is considered a significant problem for veterinary medicine worldwide. The evaluation of the transmission risk of fasciolosis as well as the efficacy of the strategies for its control could be carried out through epidemiological surveillance of the snails that act as intermediate hosts of the parasites. The present study aimed to develop the first multiplex PCR to detect Fasciola hepatica in Galba cubensis, an important intermediate host of the parasite in the Americas and especially in the Caribbean basin. The multiplex PCR was optimized for the amplification of a 340 bp fragment of the second internal transcribed spacer (ITS-2) of F. hepatica rDNA, while another set of primers was designed and used to amplify a conserved segment of the nuclear 18S rDNA of the snail (451 bp), as an internal control of the reaction. The assay was able to detect up to 100 pg of the parasite even at high concentrations of snail DNA, an analytical sensitivity that allows the detection of less than a single miracidium, which is the minimal biological infestation unit. A controlled laboratory-reared G. cubensis - F. hepatica system was used for the evaluation of the developed multiplex PCR, and 100% sensitivity and specificity was achieved. This assay constitutes a novel, useful and suitable technique for the survey of fasciolosis transmission through one of the main intermediate hosts in the Western hemisphere. PMID:26012858

  2. Early Pathological Changes Associated with Fasciola hepatica Infection in White-tailed Deer

    PubMed Central

    Presidente, P. J. A.; McCraw, B. M.; Lumsden, J. H.

    1974-01-01

    Three white-tailed deer (Odocoileus virginianus) were inoculated with 1000 metacercariae of Fasciola hepatica and examined on days 7, 14, and 28 postinoculation to determine the early response of a resistant host to this infection. It was concluded that only small numbers of the metacercariae penetrated the intestinal wall into the peritoneal cavity. Flukes that migrated to the liver penetrated through Glisson's capsule, primarily on the parietal surface. Marked fibroplasia and cellular infiltration of the capsule were induced and flukes were killed and destroyed in granulomas immediately beneath the capsule. Migration in hepatic parenchyma was minimal and immature flukes or migratory tracks were not found. There were infiltrations of eosinophils and mononuclear cells, bile duct hyperplasia and fibroplasia in portal areas. A few flukes penetrated through the diaphragm within 14 days postinoculation and on day 28 granulomas were observed on the dorsal surface of the lung where F. hepatica had penetrated this organ. The early reaction of Glisson's capsule to F. hepatica infection in white-tailed deer has not been described in cattle, sheep or swine infected with this fluke. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4.Fig. 5.Fig. 6.Fig. 7.Fig. 8.Fig. 9.Fig. 10. PMID:4277445

  3. Epidemiology and impact of Fasciola hepatica exposure in high-yielding dairy herds.

    PubMed

    Howell, Alison; Baylis, Matthew; Smith, Rob; Pinchbeck, Gina; Williams, Diana

    2015-09-01

    The liver fluke Fasciola hepatica is a trematode parasite with a worldwide distribution and is the cause of important production losses in the dairy industry. The aim of this observational study was to assess the prevalence of exposure to F. hepatica in a group of high yielding dairy herds, to determine the risk factors and investigate their associations with production and fertility parameters. Bulk milk tank samples from 606 herds that supply a single retailer with liquid milk were tested with an antibody ELISA for F. hepatica. Multivariable linear regression was used to investigate the effect of farm management and environmental risk factors on F. hepatica exposure. Higher rainfall, grazing boggy pasture, presence of beef cattle on farm, access to a stream or pond and smaller herd size were associated with an increased risk of exposure. Univariable regression was used to look for associations between fluke exposure and production-related variables including milk yield, composition, somatic cell count and calving index. Although causation cannot be assumed, a significant (p<0.001) negative association was seen between F. hepatica exposure and estimated milk yield at the herd level, representing a 15% decrease in yield for an increase in F. hepatica exposure from the 25th to the 75th percentile. This remained significant when fertility, farm management and environmental factors were controlled for. No associations were found between F. hepatica exposure and any of the other production, disease or fertility variables.

  4. Fasciola hepatica Kunitz Type Molecule Decreases Dendritic Cell Activation and Their Ability to Induce Inflammatory Responses

    PubMed Central

    Falcón, Cristian R.; Masih, Diana; Gatti, Gerardo; Sanchez, María Cecilia; Motrán, Claudia C.; Cervi, Laura

    2014-01-01

    The complete repertoire of proteins with immunomodulatory activity in Fasciola hepatica (Fh) has not yet been fully described. Here, we demonstrated that Fh total extract (TE) reduced LPS-induced DC maturation, and the DC ability to induce allogeneic responses. After TE fractionating, a fraction lower than 10 kDa (F<10 kDa) was able to maintain the TE properties to modulate the DC pro- and anti-inflammatory cytokine production induced by LPS. In addition, TE or F<10 kDa treatment decreased the ability of immature DC to stimulate the allogeneic responses and induced a novo allogeneic CD4+CD25+Foxp3+ T cells. In contrast, treatment of DC with T/L or F<10 kDa plus LPS (F<10/L) induced a regulatory IL-27 dependent mechanism that diminished the proliferative and Th1 and Th17 allogeneic responses. Finally, we showed that a Kunitz type molecule (Fh-KTM), present in F<10 kDa, was responsible for suppressing pro-inflammatory cytokine production in LPS-activated DC, by printing tolerogenic features on DC that impaired their ability to induce inflammatory responses. These results suggest a modulatory role for this protein, which may be involved in the immune evasion mechanisms of the parasite. PMID:25486609

  5. Excretory-secretory products (ESP) from Fasciola hepatica induce tolerogenic properties in myeloid dendritic cells.

    PubMed

    Falcón, Cristian; Carranza, Franco; Martínez, Fernando F; Knubel, Carolina P; Masih, Diana T; Motrán, Claudia C; Cervi, Laura

    2010-09-15

    Fasciola hepatica is a helminth trematode that migrates through the host tissues until reaching bile ducts where it becomes an adult. During its migration the parasite releases different excretory-secretory products (ESP), which are in contact with the immune system. In this study, we focused on the effect of ESP on the maturation and function of murine bone marrow derived-dendritic cells (DC). We found that the treatment of DC with ESP failed to induce a classical maturation of these cells, since ESP alone did not activate DC to produce any cytokines, although they impaired the ability of DC to be activated by TLR ligands and also their capacity to stimulate an allospecific response. In addition, using an in vitro ovalbumin peptide-restricted priming assay, ESP-treated DC exhibited a capacity to drive Th2 and regulatory T cell (Treg) polarization of CD4(+) cells from DO11.10 transgenic mice. This was characterized by increased IL-4, IL-5, IL-10 and TGF-beta production and the expansion of CD4(+)CD25(+)Foxp3(+) cells. Our results support the hypothesis that ESP from F. hepatica modulate the maturation and function of DC as part of a generalized immunosuppressive mechanism that involves a bias towards a Th2 response and Treg development.

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

    PubMed

    Vignoles, Philippe; Dreyfuss, Gilles; Rondelaud, Daniel

    2015-01-01

    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. PMID:25907356

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

    PubMed

    Vignoles, Philippe; Dreyfuss, Gilles; Rondelaud, Daniel

    2015-01-01

    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.

  8. Generating a detailed protein profile of Fasciola hepatica during the chronic stage of infection in cattle.

    PubMed

    Haçarız, Orçun; Baykal, Ahmet Tarık; Akgün, Mete; Kavak, Pınar; Sağıroğlu, Mahmut Şamil; Sayers, Gearóid Patrick

    2014-06-01

    Fasciola hepatica is a trematode helminth causing a damaging disease, fasciolosis, in ruminants and humans. Comprehensive proteomic studies broaden our knowledge of the parasite's protein profile, and provide new insights into the development of more effective strategies to deal with fasciolosis. The objective of this study was to generate a comprehensive profile of F. hepatica proteins expressed during the chronic stage of infection in cattle by building on previous efforts in this area. The approach included an improved sample preparation procedure for surface and internal layers of the parasite, the application of nano-UPLC-ESI-qTOF-MS (nano-ultra-performance LC and ESI quadrupole TOF MS) integrated with different acquisition methods and in silico database search against various protein databases and a transcript database including a new assembly of publically available EST. Of a total of 776 identified proteins, 206 and 332 were specific to the surface and internal layers of the parasite, respectively. Furthermore, 238 proteins were common to both layers, with comparative differences of 172 proteins detected. Specific proteins not previously identified in F. hepatica, but shown to be immunomodulatory or potential drug targets for other parasites, are discussed.

  9. Fasciola hepatica in goats from north-western Spain: Risk factor analysis using a capture ELISA.

    PubMed

    Pérez-Creo, Ana; Díaz, Pablo; López, Ceferino; Béjar, Juan Pablo; Martínez-Sernández, Victoria; Panadero, Rosario; Díez-Baños, Pablo; Ubeira, Florencio M; Morrondo, Patrocinio

    2016-02-01

    In order to study the seroprevalence of Fasciola hepatica infection in goats from north-western Spain, a total of 603 serum samples from 47 herds were tested using a capture ELISA (MM3-SERO). The identification of risk factors was assessed by a mixed-effects logistic regression analysis. The results showed that F. hepatica is widespread in this area with 57.4% of the herds and 22.7% of the animals testing positive. Breed and age were identified as determining factors for caprine F. hepatica infection. Seroprevalence in cross-bred animals was significantly higher than in the autochthonous Cabra Galega breed. A significantly higher seroprevalence was observed in older animals. The use of locally adapted breeds and the implementation of suitable management practices could provide a substantial improvement over the current F. hepatica control measures carried out in goat herds and should be considered when designing new F. hepatica control programs.

  10. Lymnaea glabra: progressive increase in susceptibility to Fasciola hepatica through successive generations of experimentally infected snails.

    PubMed

    Rondelaud, D; Teukeng, F F Djuikwo; Vignoles, P; Dreyfuss, G

    2015-07-01

    Experimental infections of Lymnaea glabra (two populations) with Fasciola hepatica were carried out during seven successive snail generations, to determine if prevalence and intensity of snail infection increased over time through descendants of snails already infected with F. hepatica. Controls were descendants coming from uninfected parents and infected according to the same protocol. No larval forms were found in the bodies of control snails coming from uninfected parents. In contrast, prevalence and intensity of F. hepatica infection in snails originating from infected parents progressively increased from the F2 or F3 to the F6 generation of L. glabra. In another experiment carried out with the F7 generations of L. glabra and a single generation of Galba truncatula (as controls), the prevalence of F. hepatica infection and the total number of cercariae were lower in L. glabra (without significant differences between both populations). If the number of cercariae shed by infected snails was compared to overall cercarial production noted in snails containing cercariae but dying without emission, the percentage was greater in G. truncatula (69% instead of 52-54% in L. glabra). Even if most characteristics of F. hepatica infection were lower in L. glabra, prevalence and intensity of parasite infection increased with snail generation when tested snails came from infected parents. This mode of snail infection with F. hepatica suggests an explanation for cases of fasciolosis occurring in cattle-breeding farms where paramphistomosis is lacking and G. truncatula is absent.

  11. Study of surface carbohydrates in Galba truncatula tissues before and after infection with Fasciola hepatica.

    PubMed

    Georgieva, Katya; Georgieva, Liliya; Mizinska-Boevska, Yana; Stoitsova, Stoyanka R

    2016-07-01

    The presence and distribution of surface carbohydrates in the tissues of Galba truncatula snails uninfected or after infection with Fasciola hepatica as well as on the surface of the snail-pathogenic larval stages of the parasite were studied by lectin labelling assay. This is an attempt to find similarities that indicate possible mimicry, utilised by the parasite as an evasion strategy in this snail-trematode system. Different binding patterns were identified on head-foot-mantle, hepatopancreas, genital glands, renopericardial complex of the host as well as of the snail-pathogenic larval stages of F. hepatica. The infection with F. hepatica leads to changes of labelling with Glycine max in the head-mantle cells and Arachis hypogaea in the tubular epithelium of the hepatopancreas. The lectin binding on the other snail tissues is not changed by the development of the larvae. Our data clearly demonstrated the similarity in labelling of G. truncatula tissues and the surface of the snail-pathogenic larval stages of F. hepatica. The role of glycosylation of the contact surfaces of both organisms in relation to the host-parasite interactions is also discussed.

  12. Fasciola hepatica in snails collected from water-dropwort fields using PCR.

    PubMed

    Kim, Hwang-Yong; Choi, In-Wook; Kim, Yeon-Rok; Quan, Juan-Hua; Ismail, Hassan Ahmed Hassan Ahmed; Cha, Guang-Ho; Hong, Sung-Jong; Lee, Young-Ha

    2014-12-01

    Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea.

  13. A multiplex PCR for the detection of Fasciola hepatica in the intermediate snail host Galba cubensis.

    PubMed

    Alba, Annia; Vázquez, Antonio A; Hernández, Hilda; Sánchez, Jorge; Marcet, Ricardo; Figueredo, Mabel; Sarracent, Jorge; Fraga, Jorge

    2015-07-30

    Fasciolosis is a snail-borne trematode infection that has re-emerged as a human disease, and is considered a significant problem for veterinary medicine worldwide. The evaluation of the transmission risk of fasciolosis as well as the efficacy of the strategies for its control could be carried out through epidemiological surveillance of the snails that act as intermediate hosts of the parasites. The present study aimed to develop the first multiplex PCR to detect Fasciola hepatica in Galba cubensis, an important intermediate host of the parasite in the Americas and especially in the Caribbean basin. The multiplex PCR was optimized for the amplification of a 340 bp fragment of the second internal transcribed spacer (ITS-2) of F. hepatica rDNA, while another set of primers was designed and used to amplify a conserved segment of the nuclear 18S rDNA of the snail (451 bp), as an internal control of the reaction. The assay was able to detect up to 100 pg of the parasite even at high concentrations of snail DNA, an analytical sensitivity that allows the detection of less than a single miracidium, which is the minimal biological infestation unit. A controlled laboratory-reared G. cubensis - F. hepatica system was used for the evaluation of the developed multiplex PCR, and 100% sensitivity and specificity was achieved. This assay constitutes a novel, useful and suitable technique for the survey of fasciolosis transmission through one of the main intermediate hosts in the Western hemisphere.

  14. Exploring and Expanding the Fatty-Acid-Binding Protein Superfamily in Fasciola Species.

    PubMed

    Morphew, Russell M; Wilkinson, Toby J; Mackintosh, Neil; Jahndel, Veronika; Paterson, Steve; McVeigh, Paul; Abbas Abidi, Syed M; Saifullah, Khalid; Raman, Muthusamy; Ravikumar, Gopalakrishnan; LaCourse, James; Maule, Aaron; Brophy, Peter M

    2016-09-01

    The liver flukes Fasciola hepatica and F. gigantica infect livestock worldwide and threaten food security with climate change and problematic control measures spreading disease. Fascioliasis is also a foodborne disease with up to 17 million humans infected. In the absence of vaccines, treatment depends on triclabendazole (TCBZ), and overuse has led to widespread resistance, compromising future TCBZ control. Reductionist biology from many laboratories has predicted new therapeutic targets. To this end, the fatty-acid-binding protein (FABP) superfamily has proposed multifunctional roles, including functions intersecting vaccine and drug therapy, such as immune modulation and anthelmintic sequestration. Research is hindered by a lack of understanding of the full FABP superfamily complement. Although discovery studies predicted FABPs as promising vaccine candidates, it is unclear if uncharacterized FABPs are more relevant for vaccine formulations. We have coupled genome, transcriptome, and EST data mining with proteomics and phylogenetics to reveal a liver fluke FABP superfamily of seven clades: previously identified clades I-III and newly identified clades IV-VII. All new clade FABPs were analyzed using bioinformatics and cloned from both liver flukes. The extended FABP data set will provide new study tools to research the role of FABPs in parasite biology and as therapy targets. PMID:27495901

  15. The definitive and intermediate hosts of Fasciola hepatica in the natural watercress beds in central France.

    PubMed

    Rondelaud, D; Vignoles, P; Abrous, M; Dreyfuss, G

    2001-06-01

    Field investigations were carried out over a two-year period in 52 natural watercress beds located in the Limousin region of central France to list the mammal and bird species that frequented these sites. This enabled detection of the definitive hosts of Fasciola hepatica and determination of the prevalence of natural infection in snails. A total of 13 mammal and five bird species were listed in these watercress beds. Adult flukes were found in Lepus capensis (39.2%), Oryctolagus cuniculus (42.0%), and Sylvilagus floridanus (25.0%). No infection with F. hepatica was noted in the five species of rodents studied. Snails infected with F. hepatica were found in 14 watercress beds. The global prevalence of natural infection was 1.1% in Lymnaea truncatula and 0.3% in L. glabra. Among the other trematode larval forms detected, the most frequent was Haplometra cylindracea (0.5%). In the Limousin region, the presence of hares and rabbits in watercress beds ensured the continuation of the F. hepatica life cycle and permitted the subsequent infection of humans when this wild watercress was eaten.

  16. Fasciola hepatica: epidemiological surveillance of natural watercress beds in central France.

    PubMed

    Dreyfuss, G; Vignoles, P; Rondelaud, D

    2005-03-01

    A total of 59 natural watercress beds in the Limousin region (central France) was surveyed over a 15-year period (1990-2004) to detect the contamination of watercress by the metacercariae of Fasciola hepatica and to determine the presence of larval forms in the two species of lymnaeids which live in these waterholes in June and July. The number of beds contaminated with F. hepatica metacercariae varied over the years, and the burden of the larvae on plants was low: a mean of 2.6-6.3 per bed. The same variability was also noted for natural infections of Galba truncatula with F. hepatica, as the annual prevalences ranged from 1.2% to 2.4%. Natural infections of Omphiscola glabra with F. hepatica were only detected from 1996 and the annual prevalences subsequently increased up to 1.4-1.8% between 2001 and 2004. However, for both lymnaeids, the variations in these prevalences with year were insignificant. The contamination of these beds with F. hepatica over the past 15 years was similar to that recorded in the same sites between 1970 and 1986. The main changes were the appearance of another digenea, Paramphistomum daubneyi, in the beds, and the possibility for O. glabra to naturally sustain the larval development of F. hepatica.

  17. Histopathological and immunohistochemical study of lambs experimentally infected with Fasciola hepatica and Schistosoma bovis.

    PubMed

    Ferreras, M C; García-Iglesias, M J; Manga-González, M Y; Pérez-Martínez, C; Mizinska, Y; Ramajo, V; González-Lanza, M C; Escudero, A; García-Marín, J F

    2000-12-01

    The aim of this study was to investigate the cross-resistance between Fasciola hepatica and Schistosoma bovis in lambs assessing parasitologic, gross pathologic, histopathologic and immunohistochemical changes in liver and small intestine. Thirty Castellana breed lambs were divided into five comparable groups and exposed to F. hepatical S. bovis (group F/S), S. bovis/F. hepatica (group S/F), S. bovis (group S) or F. hepatica (group F) and six unexposed lambs were used as non-infected controls (group C). Primary patent infection with F. hepatica induced a lower number of schistosome eggs and a higher number of lymphocytes in intestinal and liver schistosome egg-induced granulomas in group F/S than in the groups S/F and S, liver damage being mainly attributed to F. hepatica. S. bovis infection followed by challenge with F. hepatica particularly increased the severity of the most significant liver alterations (cholangiohepatitis by F. hepatica and mesoendophlebitis by S. bovis) and F. hepatica seemed not to have an influence on established S. bovis infection. In addition, immunohistochemical results suggested that the predominant local immune response in both double-infected groups was different, being mainly a cell-mediated immune response in group F/S and a mucosal response in group S/F.

  18. Generating a detailed protein profile of Fasciola hepatica during the chronic stage of infection in cattle.

    PubMed

    Haçarız, Orçun; Baykal, Ahmet Tarık; Akgün, Mete; Kavak, Pınar; Sağıroğlu, Mahmut Şamil; Sayers, Gearóid Patrick

    2014-06-01

    Fasciola hepatica is a trematode helminth causing a damaging disease, fasciolosis, in ruminants and humans. Comprehensive proteomic studies broaden our knowledge of the parasite's protein profile, and provide new insights into the development of more effective strategies to deal with fasciolosis. The objective of this study was to generate a comprehensive profile of F. hepatica proteins expressed during the chronic stage of infection in cattle by building on previous efforts in this area. The approach included an improved sample preparation procedure for surface and internal layers of the parasite, the application of nano-UPLC-ESI-qTOF-MS (nano-ultra-performance LC and ESI quadrupole TOF MS) integrated with different acquisition methods and in silico database search against various protein databases and a transcript database including a new assembly of publically available EST. Of a total of 776 identified proteins, 206 and 332 were specific to the surface and internal layers of the parasite, respectively. Furthermore, 238 proteins were common to both layers, with comparative differences of 172 proteins detected. Specific proteins not previously identified in F. hepatica, but shown to be immunomodulatory or potential drug targets for other parasites, are discussed. PMID:24733753

  19. Characteristics of Fasciola hepatica infections in Galba truncatula originating from riverbank populations.

    PubMed

    Vignoles, P; Rondelaud, D; Dreyfuss, G

    2011-03-01

    Four geographical strains of Galba truncatula living on riverbanks (the first on a sedimentary soil and the other three on an acid soil) were subjected to bimiracidial exposures with Fasciola hepatica to study their aptitude for cercarial shedding and to count metacercariae in snails dissected at day 42 post-exposure. All snails were reared in 14-cm Petri dishes at 24 °C, with the same spring water (60-73 mg/l of Ca²+) and the same diet (grass and lettuce leaves). Metacercariae of F. hepatica were noted in the four populations after a cercarial shedding or after snail dissection. However, in spite of the breeding method used, the characteristics of snail infections varied with the origin of each geographical strain. For example, the shell heights of infected snails at day 42 were close to those found for the corresponding adults in the field (6.8-8.0 mm for the population living on the sedimentary soil, but only 4.6-5.5 mm for another strain originating from the acid soil). This variability may be explained by assuming that the diet of these riverbank G. truncatula would be different from that of snails living in swampy meadows. However, another hypothesis based on the influence of snail habitat on the characteristics of snail life cannot be excluded. PMID:20388248

  20. Fasciola hepatica tegumental antigens indirectly induce an M2 macrophage-like phenotype in vivo.

    PubMed

    Adams, P N; Aldridge, A; Vukman, K V; Donnelly, S; O'Neill, S M

    2014-10-01

    The M2 subset of macrophages has a critical role to play in host tissue repair, tissue fibrosis and modulation of adaptive immunity during helminth infection. Infection with the helminth, Fasciola hepatica, is associated with M2 macrophages in its mammalian host, and this response is mimicked by its excretory-secretory products (FhES). The tegumental coat of F. hepatica (FhTeg) is another major source of immune-modulatory molecules; we have previously shown that FhTeg can modulate the activity of both dendritic cells and mast cells inhibiting their ability to prime a Th1 immune response. Here, we report that FhTeg does not induce Th2 immune responses but can induce M2-like phenotype in vivo that modulates cytokine production from CD4(+) cells in response to anti-CD3 stimulation. FhTeg induces a RELMα expressing macrophage population in vitro, while in vivo, the expression of Arg1 and Ym-1/2 but not RELMα in FhTeg-stimulated macrophages was STAT6 dependent. To support this finding, FhTeg induces RELMα expression in vivo prior to the induction of IL-13. FhTeg can induce IL-13-producing peritoneal macrophages following intraperitoneal injection This study highlights the important role of FhTeg as an immune-modulatory source during F. hepatica infection and sheds further light on helminth-macrophage interactions.

  1. Oxidative stress associated with pathological lesions in the liver of rats experimentally infected by Fasciola hepatica.

    PubMed

    Bottari, Nathieli B; Mendes, Ricardo E; Lucca, Neuber J; Schwertz, Claiton I; Henker, Luan C; Olsson, Débora C; Piva, Manoela M; Sangoi, Manuela; Campos, Luízi P; Moresco, Rafael N; Jaques, Jeandre A; Da silva, Aleksandro S

    2015-12-01

    The aim of this study was to analyze the antioxidant status and oxidative profile in serum and liver of rats experimentally infected with Fasciola hepatica and its relationship with pathological findings. Twenty-four rats were divided into two groups: group A consisted of 12 healthy rats and group B of 12 rats infected orally with 20 metacercaria of F. hepatica. At days 20 and 150 post-infection (PI), blood and liver samples of six animals from each group were collected. The protein oxidation (AOPP technique: advanced oxidation protein products) and antioxidants (FRAP technique: ferric reducing antioxidant power) levels were measured in serum and liver. Furthermore, nitrite/nitrate (NOx) levels and lipid peroxidation (TBARS technique: thiobarbituric acid reactive substances) were measured in liver. AOPP and FRAP levels were increased (P < 0.05) in serum and liver of infected animals in acute and chronic infection when compared with healthy animals. The same occurred with TBARS and NOx levels in the liver (P < 0.05). Histopathology revealed periportal fibrous hepatitis, composed of an abundant inflammatory infiltrate in portal spaces on infected animals, as well as bile duct hyperplasia. The results found seem to be related to the host free radical production demonstrated in serum samples and liver due to the parasite infection.

  2. Preferential regeneration of thioredoxin from parasitic flatworm Fasciola gigantica using glutathione system.

    PubMed

    Gupta, Ankita; Pandey, Tripti; Kumar, Bijay; Tripathi, Timir

    2015-11-01

    The maintenance of cellular redox homeostasis is a crucial adaptive problem faced by parasites, and its disruption can shift the biochemical balance toward the host. The thioredoxin (Trx) system plays a key role in redox metabolism and defense against oxidative stress. In this study, biochemical experiments were performed on Fasciola gigantica Thioredoxin1 (FgTrx1). The recombinant FgTrx1 exists as a monomer and catalyzes the reduction of insulin. FgTrx1 is preferentially regenerated by the glutathione (GSH) system using glutathione reductase (GR). The regeneration of FgTrx1 by the conventional Trx system is much less as compared to the GSH system, suggesting that FgTrx1 could be acting as glutaredoxin (Grx). DNA nicking and hydroperoxide assay suggests that it protects the DNA from radical-induced oxidative damage. Thus, FgTrx1 might play a role in parasite survival as it can regenerate itself even in the absence of the canonical Trx system and also protect the cells from ROS induced damage. Further, we propose that the GR activity of FgTrx1 is not restricted to -CXXC- motif but is regulated by residues present in close proximity to the -CXXC- motif, through manipulation of the redox potential or the pKa of the active site Cys residues.

  3. Across intra-mammalian stages of the liver f luke Fasciola hepatica: a proteomic study.

    PubMed

    Di Maggio, Lucía Sánchez; Tirloni, Lucas; Pinto, Antonio F M; Diedrich, Jolene K; Yates Iii, John R; Benavides, Uruguaysito; Carmona, Carlos; da Silva Vaz, Itabajara; Berasain, Patricia

    2016-01-01

    Fasciola hepatica is the agent of fasciolosis, a foodborne zoonosis that affects livestock production and human health. Although flukicidal drugs are available, re-infection and expanding resistance to triclabendazole demand new control strategies. Understanding the molecular mechanisms underlying the complex interaction with the mammalian host could provide relevant clues, aiding the search for novel targets in diagnosis and control of fasciolosis. Parasite survival in the mammalian host is mediated by parasite compounds released during infection, known as excretory/secretory (E/S) products. E/S products are thought to protect parasites from host responses, allowing them to survive for a long period in the vertebrate host. This work provides in-depth proteomic analysis of F. hepatica intra-mammalian stages, and represents the largest number of proteins identified to date for this species. Functional classification revealed the presence of proteins involved in different biological processes, many of which represent original findings for this organism and are important for parasite survival within the host. These results could lead to a better comprehension of host-parasite relationships, and contribute to the development of drugs or vaccines against this parasite. PMID:27600774

  4. Across intra-mammalian stages of the liver f luke Fasciola hepatica: a proteomic study

    PubMed Central

    Di Maggio, Lucía Sánchez; Tirloni, Lucas; Pinto, Antonio F. M.; Diedrich, Jolene K.; Yates III, John R.; Benavides, Uruguaysito; Carmona, Carlos; da Silva Vaz Jr., Itabajara; Berasain, Patricia

    2016-01-01

    Fasciola hepatica is the agent of fasciolosis, a foodborne zoonosis that affects livestock production and human health. Although flukicidal drugs are available, re-infection and expanding resistance to triclabendazole demand new control strategies. Understanding the molecular mechanisms underlying the complex interaction with the mammalian host could provide relevant clues, aiding the search for novel targets in diagnosis and control of fasciolosis. Parasite survival in the mammalian host is mediated by parasite compounds released during infection, known as excretory/secretory (E/S) products. E/S products are thought to protect parasites from host responses, allowing them to survive for a long period in the vertebrate host. This work provides in-depth proteomic analysis of F. hepatica intra-mammalian stages, and represents the largest number of proteins identified to date for this species. Functional classification revealed the presence of proteins involved in different biological processes, many of which represent original findings for this organism and are important for parasite survival within the host. These results could lead to a better comprehension of host-parasite relationships, and contribute to the development of drugs or vaccines against this parasite. PMID:27600774

  5. [Natural infestation of Lymnaea truncatula Muller by Fasciola hepatica in the Tozeur oasis in southwest Tunisia].

    PubMed

    Hammami, H; Ayadi, A

    2000-01-01

    Natural infestation of Lymnaea truncatula by Fasciola hepatica was studied at various sites in the traditional oasis of Tozeur 8 times between September 1997 and August 1998. Infestation of snails was documented in five of the eight sites with a mean level of 26.1 p. 100. Level of infestation varied depending on the site. It was highest in secondary irrigation canals (32.8 p. 100), seguias, (34.8 p. 100), and secondary drainage canals (34.8 p. 100). Lower levels were observed in the main drainage canal, (15.2 p. 100). Two infestation periods were noted at two sites in relation with spreading of manure by farmers in autumn and spring. Perennial infestation due to the permanent presence of animal hosts was noted at one. Infestation of drainage canals is enhanced by incoming parasite eggs from upstream irrigation canals. Infestation was age dependent reaching up to 100 p. 100 in older snails of 5.5 mm. Environmental factors such as climatic conditions and human intervention are important to sustaining snail infestation in the oasis of Tozeur.

  6. [Geopolitical distribution of the occurrence of Fasciola hepatica in Santa Catarina State, Brazil].

    PubMed

    da Serra-Freire, N M; Nuernberg, S

    1992-01-01

    During 12 years feces samples from cows, water buffaloes, sheeps and goats were examined by sequential tamis filtration to show the occurrence of Fasciola hepatica eggs. The material came from 129 municipalities of Santa Catarina State, and 5 g of feces per animal were examined. The occurrence of F. hepatica was confirmed in 64.82% of the municipalities. Considering the host, F. hepatica was confirmed in goats from Florianópolis, São José, São João Batista and Guaramirim municipalities; in sheep from Brusque, Pomerode, Palhoça and São José; in water buffaloes from 9 and in cows from 86 municipalities. For this study, 13,762 feces samples were examined and in 3,814 the presence of eggs of F. hepatica was demonstrated. The percentage of occurrence for host species was 27.86 in cows, 24.72 in water buffaloes, 16.92 in sheep and 15.66 in goats. By the results it was demonstrated that Itajaí Valley at Southeast Hidrographic Basin, in Santa Catarina State is an endemic area of F. hepatica, even though Uruguai Hidrographic Basin was not referred as a geographical record for this parasite.

  7. Metacercarial aggregation in Digenea (Fasciola hepatica and Paramphistomum daubneyi): environmental or species determinism?

    PubMed

    Abrous, M; Vareille-Morel, C; Rondelaud, D; Dreyfuss, G; Cabaret, J

    2001-12-01

    Metacercarial aggregation of Fasciola hepatica and Paramphistomum daubneyi was studied under experimental conditions to determine if the formation of these aggregates was influenced by environmental factors, or it was a characteristic of trematode species. This process was studied using the confinement of infected snails on the bottom of Petri dishes (diameter, 14 cm) for 3 days. The formation of metacercarial aggregates of F. hepatica was not significantly modified by environmental factors such as intensity and duration of lighting, quality and volume of water. Metacercariae of F. hepatica were more numerous on the Petri dish walls and 63.9% of them constituted aggregates. In contrast, most metacercariae of P. daubneyi were found on the Petri dish bottoms and 78.3% of them were isolated or in groups of two metacercariae each. The mean number of metacercariae per aggregate ranged from 6.7 to 12.2 in the case of F. hepatica, and from 2.7 to 4.5 in the case of P. daubneyi. However, these mean numbers were independent of the site of cercarial attachment. The tendency of cercariae to form metacercarial aggregations was a characteristic of F. hepatica and was species determined. PMID:11818045

  8. Zoonotic potential of infection with Fasciola spp. by consumption of freshly prepared raw liver containing immature flukes.

    PubMed

    Taira, N; Yoshifuji, H; Boray, J C

    1997-07-01

    Mice were successfully infected with metacercariae of the Japanese Fasciola sp., resulting in the recovery of a mean number of 110 live immature flukes per mouse 4-5 days after inoculation. Twenty-four mice were then inoculated orally, each with a mean number of 68 freshly recovered immature flukes. The livers of 7 of the 24 recipient mice showed migratory lesions of capsular and subcapsular granulomatous infiltration and 2 of those mice also had haemorrhagic lesions. The lesions were typical of those caused by active migration of early immature flukes. However, no flukes were found in the livers of the recipient mice at necropsy when the flukes were aged 14 weeks. In another experiment, 10 piglets were given fresh livers of mice harbouring approximately 2000 live immature flukes aged 3-7 days. Two additional piglets were inoculated with 2000 metacercariae of Fasciola. All pigs were killed when the flukes were 14 days old. Granulomatous lesions were present in all pigs, except in those that were given livers containing flukes aged 7 days. The lesions were localized, forming well-defined foci, different from the typical migratory lesions normally observed in mouse or sheep liver at the early stage of fluke migration. From the 10 pigs given livers, 65 live flukes were recovered at necropsy, 0.29% of the estimated number of immature flukes given. From the 2 pigs which received 2000 metacercariae each, a total of 198 flukes were recovered (5%). The results of the experiments suggest that humans consuming raw liver dishes prepared from fresh livers infected with immature Fasciola spp. could become infected with liver fluke. PMID:9279579

  9. The effect of lysergic acid diethylamide, 5-hydroxytryptamine, and related compounds on the liver fluke, Fasciola hepatica.

    PubMed

    MANSOUR, T E

    1957-12-01

    The rhythmical activity of the liver fluke, Fasciola hepatica, was stimulated by 5-hydroxytryptamine and by lysergic acid diethylamide at very low concentrations. The effect was peripheral and was not mediated through the central ganglion. Other amines also stimulated rhythmical activity, the most potent being the indolamines.Bromolysergic acid diethylamide, and other analogues such as yohimbine, harmine, and dopamine depressed rhythmical movement and antagonized the stimulant action of 5-hydroxytryptamine and lysergic acid diethylamide. Evidence which suggests the presence of tryptamine receptors in the trematode is discussed.

  10. DNA sequence analyses reveal co-occurrence of novel haplotypes of Fasciola gigantica with F. hepatica in South Africa and Zimbabwe.

    PubMed

    Mucheka, Vimbai T; Lamb, Jennifer M; Pfukenyi, Davies M; Mukaratirwa, Samson

    2015-11-30

    The aim of this study was to identify and determine the genetic diversity of Fasciola species in cattle from Zimbabwe, the KwaZulu-Natal and Mpumalanga provinces of South Africa and selected wildlife hosts from Zimbabwe. This was based on analysis of DNA sequences of the nuclear ribosomal internal transcribed spacer (ITS1 and 2) and mitochondrial cytochrome oxidase 1 (CO1) regions. The sample of 120 flukes was collected from livers of 57 cattle at 4 abattoirs in Zimbabwe and 47 cattle at 6 abattoirs in South Africa; it also included three alcohol-preserved duiker, antelope and eland samples from Zimbabwe. Aligned sequences (ITS 506 base pairs and CO1 381 base pairs) were analyzed by neighbour-joining, maximum parsimony and Bayesian inference methods. Phylogenetic trees revealed the presence of Fasciola gigantica in cattle from Zimbabwe and F. gigantica and Fasciola hepatica in the samples from South Africa. F. hepatica was more prevalent (64%) in South Africa than F. gigantica. In Zimbabwe, F. gigantica was present in 99% of the samples; F. hepatica was found in only one cattle sample, an antelope (Hippotragus niger) and a duiker (Sylvicapra grimmia). This is the first molecular confirmation of the identity Fasciola species in Zimbabwe and South Africa. Knowledge on the identity and distribution of these liver flukes at molecular level will allow disease surveillance and control in the studied areas.

  11. ANTHELMINTIC ACTIVITY OF CHLOROPHYLLIN AGAINST DIFFERENT LARVAL STAGES OF Fasciola gigantica.

    PubMed

    Singh, Divya Jyoti; Singh, Dinesh Kumar

    2016-01-01

    Fasciolosis is a food borne zoonosis, caused by the digenetic trematode Fasciola. Freshwater lymnaeid snails are the intermediate host of the trematodes. Chlorophyllin, a semi-synthetic derivative of chlorophyll and its formulations obtained from freeze dried cow urine (FCU) had their toxicity tested against redia and cercaria larvae of F. gigantica. The larvicidal activity of chlorophyllin and its formulations were found to depend on both, time and concentration used against the larvae. Toxicity of chlorophyllin + FCU (1:1 ratio) in sunlight against redia larva (8 h LC50: 0.03 mg/mL) was more pronounced than using just chlorophyllin (8 h LC50: 0.06 mg/mL). Toxicity of chlorophyllin + FCU in sunlight against redia (8 h LC50: 0.03 mg/mL) was higher than against cercaria (8 h LC50: 0.06 mg/mL). The larvicidal activity of chlorophyllin in sunlight (redia/cercaria larvae: 8 h LC50: 0.06 mg/mL) was more pronounced than under laboratory conditions (redia: 8 h LC50: 22.21 mg/mL/, cercaria 8 h LC50: 96.21 mg/mL). Toxicity of FCU against both larvae was lower than that of chlorophyllin and chlorophyllin + FCU. Chlorophyllin and its formulations + FCU were 357.4 to 1603.5 times more effective against redia/cercaria larvae in sunlight than under laboratory conditions. The present study has shown that chlorophyllin formulations may be used as potent larvicides against fasciolosis.

  12. Fasciola hepatica Surface Tegument: Glycoproteins at the Interface of Parasite and Host*

    PubMed Central

    Ravidà, Alessandra; Cwiklinski, Krystyna; Aldridge, Allison M.; Clarke, Paul; Thompson, Roisin; Gerlach, Jared Q.; Kilcoyne, Michelle; Hokke, Cornelis H.; Dalton, John P.; O'Neill, Sandra M.

    2016-01-01

    Fasciola hepatica, commonly known as liver fluke, is a trematode that causes Fasciolosis in ruminants and humans. The outer tegumental coat of F. hepatica (FhTeg) is a complex metabolically active biological matrix that is continually exposed to the host immune system and therefore makes a good vaccine target. F. hepatica tegumental coat is highly glycosylated and helminth-derived immunogenic oligosaccharide motifs and glycoproteins are currently being investigated as novel vaccine candidates. This report presents the first systematic characterization of FhTeg glycosylation using lectin microarrays to characterize carbohydrates motifs present, and lectin histochemistry to localize these on the F. hepatica tegument. We discovered that FhTeg glycoproteins are predominantly oligomannose oligosaccharides that are expressed on the spines, suckers and tegumental coat of F. hepatica and lectin blot analysis confirmed the abundance of N- glycosylated proteins. Although some oligosaccharides are widely distributed on the fluke surface other subsets are restricted to distinct anatomical regions. We selectively enriched for FhTeg mannosylated glycoprotein subsets using lectin affinity chromatography and identified 369 proteins by mass spectrometric analysis. Among these proteins are a number of potential vaccine candidates with known immune modulatory properties including proteases, protease inhibitors, paramyosin, Venom Allergen-like II, Enolase and two proteins, nardilysin and TRIL, that have not been previously associated with F. hepatica. Furthermore, we provide a comprehensive insight regarding the putative glycosylation of FhTeg components that could highlight the importance of further studies examining glycoconjugates in host-parasite interactions in the context of F. hepatica infection and the development of an effective vaccine. PMID:27466253

  13. Immunization with Fasciola hepatica thioredoxin glutathione reductase failed to confer protection against fasciolosis in cattle.

    PubMed

    Maggioli, Gabriela; Bottini, Gualberto; Basika, Tatiana; Alonzo, Pablo; Salinas, Gustavo; Carmona, Carlos

    2016-07-15

    The liver fluke Fasciola hepatica remains an important agent of food-borne trematode disease producing great economic losses due to its negative effect on productivity of livestock grazing in temperate areas. The prevailing control strategy based on anthelmintic drugs is unsustainable due to widespread resistance hence vaccination appears as an attractive option to pursue. In this study we evaluate the effect of vaccination in calves with a functional recombinant thioredoxin glutathione reductase (rFhTGR) from liver fluke, a critical antioxidant enzyme at the crossroads of the thioredoxin and glutathione metabolism in flatworms. The recombinant enzyme produced in Escherichia coli was tested in two vaccination experiments; in the first trial rFhTGR was administered in combination with Freund́s Incomplete Adjuvant (FIA) in a three-inoculation scheme on weeks 0, 4 and 8; in the second trial rFhTGR was given mixed with Adyuvac 50 or Alum as adjuvants on weeks 0 and 4. In both cases calves were challenged with metacercariae (400 in the first and 500 in the second trial) 2 weeks after the last inoculation. Our results demonstrate that two or three doses of the vaccine induced a non-significant reduction in worm counts of 8.2% (FIA), 3.8% (Adyuvac 50) and 23.0% (Alum) compared to adjuvant controls indicating that rFhTGR failed to induce a protective immunity in challenged calves. All vaccine formulations induced a mixed IgG1/IgG2 response but no booster was observed after challenge. No correlations between antibody titres and worm burdens were found. PMID:27270384

  14. Effects of Mirazid® and myrrh volatile oil on adult Fasciola gigantica under laboratory conditions

    PubMed Central

    Massoud, AM; Shalaby, HA; El Khateeb, RM; Mahmoud, MS; Kutkat, MA

    2012-01-01

    Objective To evaluate the effects of Mirazid® and myrrh volatile oil on adult Fasciola gigantica (F. gigantica ) under laboratory conditions. Methods The effects of oleoresin extract of myrrh (Mirazid®) and myrrh volatile oil on the surface morphology of adult F. gigantica following treatment in vitro had been determined by scanning electron microscopy. The results were compared with those observed in the fluke tegument following incubation in triclabendazole sulphoxide (TCBZ-SO), active form, (Fasinex®, Ciba-Geigy). Results Observations of the efficacy of Mirazid® oleoresin extract and myrrh volatile oil indicated that both products showed dose-dependent anthelmintic efficacy. The anterior half of the fluke was consistently more severely affected than the posterior half. The surface changes induced by Mirazid® oleoresin extract were less severe than those observed after exposure to either myrrh volatile oil or TCBZ-SO. Flukes showed swelling after these treatments, but its level and blebbing were much greater with myrrh volatile oil; in which patches of tegumental sloughing were observed in the apical cone and the posterior mid-body region of flukes. This was not observed after treatment with Mirazid® oleoresin extract. Conclusions The comparatively more disruption, observed in myrrh volatile oil exposed specimens, compared to that exposed to Mirazid® oleoresin extract might suggest that the anthelmintic activity of Mirazid® oleo resin extract was attributed to its content of volatile oil. So, increasing the concentration of myrrh volatile oil in Mirazid® might possibly help to developing its anthelmintic activity. PMID:23569864

  15. Immunization with Fasciola hepatica thioredoxin glutathione reductase failed to confer protection against fasciolosis in cattle.

    PubMed

    Maggioli, Gabriela; Bottini, Gualberto; Basika, Tatiana; Alonzo, Pablo; Salinas, Gustavo; Carmona, Carlos

    2016-07-15

    The liver fluke Fasciola hepatica remains an important agent of food-borne trematode disease producing great economic losses due to its negative effect on productivity of livestock grazing in temperate areas. The prevailing control strategy based on anthelmintic drugs is unsustainable due to widespread resistance hence vaccination appears as an attractive option to pursue. In this study we evaluate the effect of vaccination in calves with a functional recombinant thioredoxin glutathione reductase (rFhTGR) from liver fluke, a critical antioxidant enzyme at the crossroads of the thioredoxin and glutathione metabolism in flatworms. The recombinant enzyme produced in Escherichia coli was tested in two vaccination experiments; in the first trial rFhTGR was administered in combination with Freund́s Incomplete Adjuvant (FIA) in a three-inoculation scheme on weeks 0, 4 and 8; in the second trial rFhTGR was given mixed with Adyuvac 50 or Alum as adjuvants on weeks 0 and 4. In both cases calves were challenged with metacercariae (400 in the first and 500 in the second trial) 2 weeks after the last inoculation. Our results demonstrate that two or three doses of the vaccine induced a non-significant reduction in worm counts of 8.2% (FIA), 3.8% (Adyuvac 50) and 23.0% (Alum) compared to adjuvant controls indicating that rFhTGR failed to induce a protective immunity in challenged calves. All vaccine formulations induced a mixed IgG1/IgG2 response but no booster was observed after challenge. No correlations between antibody titres and worm burdens were found.

  16. First report of closantel treatment failure against Fasciola hepatica in cattle.

    PubMed

    Novobilský, Adam; Höglund, Johan

    2015-12-01

    Control of Fasciola hepatica infection in livestock is based on annual treatment using flukicides such as triclabendazole, albendazole and closantel. However, triclabendazole resistant F. hepatica populations are emerging worldwide and resistance is emerging to albendazole, whereas it has until now never been described for closantel. In Sweden, a topical formulation containing a combination of closantel and ivermectin (Closamectin Pour On) has been registered for use in cattle only since 2011. This study evaluated the efficacy of closantel against F. hepatica in naturally infected beef cattle using both coproantigen and faecal egg count reduction tests. Faecal egg counts (FEC) and coproantigen ELISA examinations were conducted in February 2014 in three beef cattle herds (A, B, C) in south-western Sweden. On each farm, 10 F. hepatica coproantigen-positive and F. hepatica egg-positive animals were allocated after 12-16 weeks of housing into groups and treated topically with a minimum of 20 mg closantel per kg body weight. Faecal samples were collected from selected animals on 0, 7 and 21 day post-treatment (PT). Based on FEC, closantel efficacy 21 days PT was 72% (95% CI: 65-77%) and 97% (95% CI: 95-98%) on farms A and B, respectively. No FEC reduction at all was observed on farm C. In total, 4, 1 and 6 animals remained coproantigen-positive at 21 days PT on farms A, B and C, respectively. Closantel treatment failure was confirmed on two of the farms. As the animals were housed 12-16 weeks before treatment and thereafter during the entire study, failure due to the presence of juvenile flukes was excluded. Although the cause of closantel failure currently remains unclear, development of resistance or/and absorption failure of topical administration should be considered. To our knowledge, this is the first report of closantel treatment failure against F. hepatica in cattle.

  17. Fasciola hepatica induces eosinophil apoptosis in the migratory and biliary stages of infection in sheep.

    PubMed

    Escamilla, A; Bautista, M J; Zafra, R; Pacheco, I L; Ruiz, M T; Martínez-Cruz, S; Méndez, A; Martínez-Moreno, A; Molina-Hernández, V; Pérez, J

    2016-01-30

    The aim of the present work was to evaluate the number of apoptotic eosinophils in the livers of sheep experimentally infected with Fasciola hepatica during the migratory and biliary stages of infection. Four groups (n=5) of sheep were used; groups 1-3 were orally infected with 200 metacercariae (mc) and sacrificed at 8 and 28 days post-infection (dpi), and 17 weeks post-infection (wpi), respectively. Group 4 was used as an uninfected control. Apoptosis was detected using immunohistochemistry with a polyclonal antibody against anti-active caspase-3, and transmission electron microscopy (TEM). Eosinophils were identified using the Hansel stain in serial sections for caspase-3, and by ultrastructural features using TEM. At 8 and 28 dpi, numerous caspase-3(+) eosinophils were mainly found at the periphery of acute hepatic necrotic foci. The percentage of caspase -3(+) apoptotic eosinophils in the periphery of necrotic foci was high (46.1-53.9) at 8 and 28 dpi, respectively, and decreased in granulomas found at 28 dpi (6%). Transmission electron microscopy confirmed the presence of apoptotic eosinophils in hepatic lesions at 8 and 28 dpi. At 17 wpi, apoptotic eosinophils were detected in the infiltrate surrounding some enlarged bile ducts containing adult flukes. This is the first report of apoptosis induced by F. hepatica in sheep and the first study reporting apoptosis in eosinophils in hepatic inflammatory infiltrates in vivo. The high number of apoptotic eosinophils in acute necrotic tracts during the migratory and biliary stages of infection suggests that eosinophil apoptosis may play a role in F. hepatica survival during different stages of infection.

  18. ANTHELMINTIC ACTIVITY OF CHLOROPHYLLIN AGAINST DIFFERENT LARVAL STAGES OF Fasciola gigantica

    PubMed Central

    SINGH, Divya Jyoti; SINGH, Dinesh Kumar

    2016-01-01

    Fasciolosis is a food borne zoonosis, caused by the digenetic trematode Fasciola. Freshwater lymnaeid snails are the intermediate host of the trematodes. Chlorophyllin, a semi-synthetic derivative of chlorophyll and its formulations obtained from freeze dried cow urine (FCU) had their toxicity tested against redia and cercaria larvae of F. gigantica. The larvicidal activity of chlorophyllin and its formulations were found to depend on both, time and concentration used against the larvae. Toxicity of chlorophyllin + FCU (1:1 ratio) in sunlight against redia larva (8 h LC50: 0.03 mg/mL) was more pronounced than using just chlorophyllin (8 h LC50: 0.06 mg/mL). Toxicity of chlorophyllin + FCU in sunlight against redia (8 h LC50: 0.03 mg/mL) was higher than against cercaria (8 h LC50: 0.06 mg/mL). The larvicidal activity of chlorophyllin in sunlight (redia/cercaria larvae: 8 h LC50: 0.06 mg/mL) was more pronounced than under laboratory conditions (redia: 8 h LC50: 22.21 mg/mL/, cercaria 8 h LC50: 96.21 mg/mL). Toxicity of FCU against both larvae was lower than that of chlorophyllin and chlorophyllin + FCU. Chlorophyllin and its formulations + FCU were 357.4 to 1603.5 times more effective against redia/cercaria larvae in sunlight than under laboratory conditions. The present study has shown that chlorophyllin formulations may be used as potent larvicides against fasciolosis. PMID:27253741

  19. Unusual snail species involved in the transmission of Fasciola hepatica in watercress beds in central France.

    PubMed

    Dreyfuss, G; Vignoles, P; Abrous, M; Rondelaud, D

    2002-06-01

    Four freshwater pulmonate species (Lymnaea ovata, L. stagnalis, Physa acuta, Planorbis leucostoma) were living in several watercress beds known for their relationships with human cases of fasciolosis, whereas L. truncatula was never found. The aims of these studies were to determine the prevalence of natural infections with Fasciola hepatica in snails and to verify if these species might ensure the full larval development of this trematode (with cercarial shedding) when they were experimentally subjected to F. hepatica only, or to co-infections with an other trematode species. Investigations were so carried out in six snail populations living in watercress beds (including three for P. acuta) and in four others originating from three brooks or a pond (as controls). Snails naturally infected with F. hepatica were found in two watercress beds inhabited by L. ovata (prevalence of infection: 1.4%) and P. leucostoma (0.1%), respectively. The L. ovata from the watercress bed could be infected at a higher size than those from the control population and the prevalence of this infection was greater in the bed population. Similar findings were noted for L. stagnalis. Despite single or dual infections, the results obtained with the four populations of P. acuta were unsuccessful. In contrast, the co-infections of young P. leucostoma with Paramphistomum daubneyi and F. hepatica resulted in the shedding of some F. hepatica cercariae. According to the authors, the occurrence of fasciolosis in these watercress beds would be the consequence of frequent natural encounters between parasite and snails (L. ovata, L. stagnalis), or of co-infections with P. daubneyi and F. hepatica (P. leucostoma). In watercress beds only colonized by P. acuta, a lymnaeid species would have ensured the larval development of F. hepatica but it would have been eliminated by P. acuta, as this last species was known to be invasive and could colonize open drainage ditches on siliceous soil.

  20. ANTHELMINTIC ACTIVITY OF CHLOROPHYLLIN AGAINST DIFFERENT LARVAL STAGES OF Fasciola gigantica.

    PubMed

    Singh, Divya Jyoti; Singh, Dinesh Kumar

    2016-01-01

    Fasciolosis is a food borne zoonosis, caused by the digenetic trematode Fasciola. Freshwater lymnaeid snails are the intermediate host of the trematodes. Chlorophyllin, a semi-synthetic derivative of chlorophyll and its formulations obtained from freeze dried cow urine (FCU) had their toxicity tested against redia and cercaria larvae of F. gigantica. The larvicidal activity of chlorophyllin and its formulations were found to depend on both, time and concentration used against the larvae. Toxicity of chlorophyllin + FCU (1:1 ratio) in sunlight against redia larva (8 h LC50: 0.03 mg/mL) was more pronounced than using just chlorophyllin (8 h LC50: 0.06 mg/mL). Toxicity of chlorophyllin + FCU in sunlight against redia (8 h LC50: 0.03 mg/mL) was higher than against cercaria (8 h LC50: 0.06 mg/mL). The larvicidal activity of chlorophyllin in sunlight (redia/cercaria larvae: 8 h LC50: 0.06 mg/mL) was more pronounced than under laboratory conditions (redia: 8 h LC50: 22.21 mg/mL/, cercaria 8 h LC50: 96.21 mg/mL). Toxicity of FCU against both larvae was lower than that of chlorophyllin and chlorophyllin + FCU. Chlorophyllin and its formulations + FCU were 357.4 to 1603.5 times more effective against redia/cercaria larvae in sunlight than under laboratory conditions. The present study has shown that chlorophyllin formulations may be used as potent larvicides against fasciolosis. PMID:27253741

  1. Bioclimatic distribution and prevalence maps for Fasciola hepatica in Espírito Santo State, Brazil

    PubMed Central

    2014-01-01

    Background Fasciolosis affects different ruminant species and leads to great economic losses for cattle farmers worldwide. Thus, the current study aimed to evaluate bovine fasciolosis prevalence in the state of Espírito Santo, Brazil, using slaughter maps provided by slaughterhouses and verifying the origin of cattle. Methods A map was created based on analysis of epidemiological data. The ArcGIS/ArcINFO 10.1 software was employed in order to elaborate updated bioclimatic maps that displayed the fasciolosis prevalence within the state – per city– between 2009 and 2011. Results According to the bioclimatic map it was clear that 52.24% of the state’s total area comprise regions considered favorable for the development and survival of Fasciola hepatica. According to the data provided by slaughterhouses, the parasite was more frequent in the cities of Atílio Vivácqua, Itapemirim and Anchieta with respective prevalence of 28.41, 25.50 and 24.95%. Although the northern portion of the state is also favorable for the disease maintenance (reaching rates above 90%), several cities presented prevalence of only 0.99 and 1.94% respectively. These findings indicate that climatic and environmental factors only cannot be considered preponderant to fasciolosis occurrence. Regarding the slaughterhouse located in Anchieta city, the higher prevalence was registered in the cities of Jerônimo Monteiro, Alegre and Cachoeiro de Itapemirim, with mean prevalence of 1.21, 1.07 and 2.09% respectively. Conclusion Although the present findings suggest a pattern for the prevalence of fasciolosis, records of the cities for the occurrence of the disease usually do not reflect the true origin of animals. PMID:25101121

  2. Effect of lipopolysaccharide (LPS) from Ochrobactrum intermedium on sheep experimentally infected with Fasciola hepatica.

    PubMed

    Martínez-Pérez, J M; Robles-Pérez, D; Valcárcel-Sancho, F; González-Guirado, A M; de Castro, I Casanova-García; Nieto-Martínez, J M; Rojo-Vázquez, F A; Martínez-Valladares, M

    2013-08-01

    The effects of the lipopolysaccharide (LPS) from Ochrobactrum intermedium was evaluated in sheep experimentally infected with Fasciola hepatica. Animals were divided into four groups: two treated with the LPS (T1/T2) and two controls (C1/C2). T1/C1 were slaughtered at 30 days postinfection (dpi) and T2/C2 at 85 dpi. Body weight and body condition were found higher in T1 and T2 than in controls, although differences were not significant. Treated sheep showed lower cumulative fecal egg count than controls (p < 0.01). Levels of red blood cells (RBC), white blood cells, hemoglobin, and hematocrit (HCT) were higher in T1 and T2. Significant differences (p < 0.05) in RBC and HCT were found between groups at 84 dpi. More severe macrocytic and hypochromic anemia was observed in C1 and C2 than in treated groups. Total protein and albumin values were higher in T1 and T2 (p < 0.01) until 29 dpi. At the end of the trial, no significant differences were observed in hepatic enzymes, although gamma-glutamyl transferase and aspartate aminotransferase values were higher in C2, and alanine aminotransferase was higher in T2. At necropsy, the mean weight of liver, fibrosis in portal triads, and ganglion size were similar in all groups. The number and size of flukes was greater in C2 than in T2 (p < 0.05). The histological examinations revealed a higher degree of parenchymatous fibrosis in T2 compared to C2 (p < 0.05). The administration of LPS from O. intermedium increased the nonspecific resistance against F. hepatica in experimentally infected sheep.

  3. Pulmonary Arterial Lesions in New World Camelids in Association With Dicrocoelium dendriticum and Fasciola hepatica Infection.

    PubMed

    Hilbe, M; Robert, N; Pospischil, A; Gerspach, C

    2015-11-01

    In Switzerland, dicrocoeliasis is regarded as the most significant parasitic infection of llamas and alpacas. Fasciola hepatica infestation is also a problem but less common. The aim of the present retrospective study was to evaluate the lungs of New World camelids (NWCs) for evidence of arterial hypertension in association with liver changes due to liver fluke infestation. The lungs of 20 llamas and 20 alpacas with liver fluke infestation were histologically evaluated. The hematoxylin and eosin and van Gieson (VG)-elastica stains as well as immunohistology for the expression of α-smooth muscle actin (α-SMA) were used to visualize the structures of arterial walls. Parasitology of fecal matter (11 llamas and 17 alpacas) confirmed that most of these animals were infested with both Dicrocoelium dendriticum and other gastrointestinal parasites. In most cases (10/12 llamas, 4/6 alpacas), liver enzyme activity in serum was elevated. Histologically, arteries in the lungs of 9 of 20 llamas (45%) and 3 of 20 alpacas (15%) showed severe intimal and adventitial and slight to moderate medial thickening, which was confirmed with α-SMA and VG-elastica staining. All animals exhibited typical liver changes, such as fibrosis and biliary hyperplasia, in association with the presence of liver flukes. This study shows that liver flukes can induce proliferative changes in lung arteries in NWCs that resemble those seen with pulmonary arterial hypertension due to liver parasites in humans. However, the degree of liver fluke infestation was not correlated with the extent of liver damage, or with the amount of thoracic or abdominal effusion or pulmonary arterial changes.

  4. Distribution of Fasciola hepatica in Swedish dairy cattle and associations with pasture management factors.

    PubMed

    Novobilský, Adam; Sollenberg, Sofia; Höglund, Johan

    2015-01-01

    The geographic distribution of Fasciola hepatica infection in relation to management routines was studied in Swedish dairy herds by testing for F. hepatica antibodies with the enzyme-linked immunosorbent assay (ELISA). In addition, all farmers were sent a questionnaire asking for information about type of production, management routines and historical record of F. hepatica at slaughter. A total of 176 farmers (41%) responded to the questionnaire. A total of 426 bulk tank milk (BTM) samples were randomly selected from the period September to October 2012 representing approximately 10% of all herds in Sweden. The overall seroprevalence was 25% (n = 107; 95% confidence interval = 21-29%) with a concentration of herds located in south-western Sweden. Among the seropositive herds, 31 (29%) had antibody levels indicating production loss. There were no significant differences in seropositivity between organic and conventional herds or due to pasture management routines. The length of grazing period, which increased the risk for heifers, was found to be the most influential factor. A discrepancy was noted between reported F. hepatica presence at meat inspection and herds that were seropositive based on BTM-ELISA results. Although the largest proportion of seropositive BTM samples (80%) came from herds where liver fluke presence had been observed at meat inspection after slaughter, seropositive BTM samples were also diagnosed in five herds (17%) with no remarks at slaughter. In conclusion, F. hepatica is a common parasite in Swedish dairy herds and the month of heifer turn-out and the grazing period length were the most influential factors observed. PMID:25826310

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

    PubMed

    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. PMID:22451733

  6. Current status and its epidemiological consideration of Fasciola and Eurytrema infections in beef cattle of Japan

    PubMed Central

    OKAJIMA, Jungo; SHIBATA, Kazuhiko; TAKAHASHI, Eiichi; NAGAFUCHI, Tsuneyuki; OKAJIMA, Kazue; NONAKA, Nariaki

    2016-01-01

    To elucidate current status of fasciolosis and eurytremosis in beef cattle of Japan, inspection data of Tokyo Metropolitan Shibaura Slaughterhouse where beef cattle were shipped from all over Japan were analyzed, and questionnaire to farmers was conducted to assess the relationship between recognition of the disease occurrence in one’s own farm and attention to the diseases. The occurrence of fasciolosis and eurytremosis in beef cattle gradually decreased from 18.6% to 0.06% and from 0.58% to 0.02% during the period of 1964 to 2010, respectively. When the current data from 2009 to 2012 were analyzed, the occurrence of fasciolosis was recognized in cattle produced and fattened all over Japan, indicating the disease was prevalent nationwide. While, 97.5% of Eurytrema infection were detected in cattle produced in Okinawa, Shimane and Kagoshima, indicating the disease was endemic in these regions. Higher occurrence (>0.7%) of fasciolosis was observed in minor breeds, such as Japanese Shorthorn. Japanese Black showed 0.09% and 0.05% of occurrence for fasciolosis and eurytremosis, respectively, but F1 crossbred with Japanese Black showed lower occurrence (0.007% and 0.002%, respectively). No tendency of occurrence in the age of cattle at slaughter was recognized, indicating the infections may have occurred at the growing and early fattening stage of cattle. The questionnaire survey revealed that farmers experiencing fasciolosis had more knowledge about the disease, however, factors, such as testing parasite infections and use of anti-Fasciola dewormers, were not affected by the recognition of occurrence. PMID:26822002

  7. The Extracellular Vesicles of the Helminth Pathogen, Fasciola hepatica: Biogenesis Pathways and Cargo Molecules Involved in Parasite Pathogenesis*

    PubMed Central

    Cwiklinski, Krystyna; de la Torre-Escudero, Eduardo; Trelis, Maria; Bernal, Dolores; Dufresne, Philippe J.; Brennan, Gerard P.; O'Neill, Sandra; Tort, Jose; Paterson, Steve; Marcilla, Antonio; Dalton, John P.; Robinson, Mark W.

    2015-01-01

    Extracellular vesicles (EVs) released by parasites have important roles in establishing and maintaining infection. Analysis of the soluble and vesicular secretions of adult Fasciola hepatica has established a definitive characterization of the total secretome of this zoonotic parasite. Fasciola secretes at least two subpopulations of EVs that differ according to size, cargo molecules and site of release from the parasite. The larger EVs are released from the specialized cells that line the parasite gastrodermus and contain the zymogen of the 37 kDa cathepsin L peptidase that performs a digestive function. The smaller exosome-like vesicle population originate from multivesicular bodies within the tegumental syncytium and carry many previously described immunomodulatory molecules that could be delivered into host cells. By integrating our proteomics data with recently available transcriptomic data sets we have detailed the pathways involved with EV biogenesis in F. hepatica and propose that the small exosome biogenesis occurs via ESCRT-dependent MVB formation in the tegumental syncytium before being shed from the apical plasma membrane. Furthermore, we found that the molecular “machinery” required for EV biogenesis is constitutively expressed across the intramammalian development stages of the parasite. By contrast, the cargo molecules packaged within the EVs are developmentally regulated, most likely to facilitate the parasites migration through host tissue and to counteract host immune attack. PMID:26486420

  8. Morphological and Molecular Characterization of Lymnaeid Snails and Their Potential Role in Transmission of Fasciola spp. in Vietnam

    PubMed Central

    Doanh, Pham Ngoc; The, Dang Tat; Loan, Ho Thi; Losson, Bertrand; Caron, Yannick

    2013-01-01

    Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated. PMID:24516270

  9. The Extracellular Vesicles of the Helminth Pathogen, Fasciola hepatica: Biogenesis Pathways and Cargo Molecules Involved in Parasite Pathogenesis.

    PubMed

    Cwiklinski, Krystyna; de la Torre-Escudero, Eduardo; Trelis, Maria; Bernal, Dolores; Dufresne, Philippe J; Brennan, Gerard P; O'Neill, Sandra; Tort, Jose; Paterson, Steve; Marcilla, Antonio; Dalton, John P; Robinson, Mark W

    2015-12-01

    Extracellular vesicles (EVs) released by parasites have important roles in establishing and maintaining infection. Analysis of the soluble and vesicular secretions of adult Fasciola hepatica has established a definitive characterization of the total secretome of this zoonotic parasite. Fasciola secretes at least two subpopulations of EVs that differ according to size, cargo molecules and site of release from the parasite. The larger EVs are released from the specialized cells that line the parasite gastrodermus and contain the zymogen of the 37 kDa cathepsin L peptidase that performs a digestive function. The smaller exosome-like vesicle population originate from multivesicular bodies within the tegumental syncytium and carry many previously described immunomodulatory molecules that could be delivered into host cells. By integrating our proteomics data with recently available transcriptomic data sets we have detailed the pathways involved with EV biogenesis in F. hepatica and propose that the small exosome biogenesis occurs via ESCRT-dependent MVB formation in the tegumental syncytium before being shed from the apical plasma membrane. Furthermore, we found that the molecular "machinery" required for EV biogenesis is constitutively expressed across the intramammalian development stages of the parasite. By contrast, the cargo molecules packaged within the EVs are developmentally regulated, most likely to facilitate the parasites migration through host tissue and to counteract host immune attack.

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

    PubMed

    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.

  11. Development of a versatile in vitro method for understanding the migration of Fasciola hepatica newly excysted juveniles.

    PubMed

    Garcia-Campos, Andres; Baird, Alan W; Mulcahy, Grace

    2016-01-01

    Fasciola hepatica is a parasitic trematode that causes serious losses to livestock producers, and also zoonotic disease. The limitations of chemotherapy for the control of fasciolosis have led to significant interest in the development of vaccines to protect cattle and sheep from infection. However, relatively few studies have concentrated on the mechanisms of invasion of the gut by newly excysted juvenile liver flukes (NEJ) and the host response triggered by this event. The aim of this work was to develop an in vitro model to study invasion by NEJ, while also reducing the requirement for challenge infections of experimental animals. Fasciola hepatica metacercariae were excysted in vitro and placed into compartments containing rat distal jejunal sheets. Variations in incubation medium, chamber size and incubation temperature were used to identify optimal conditions for NEJ migration across the gut. Histological examination showed increased migration until 120 min post-incubation. The use of RPMI, without gassing at 39 °C, as the incubation medium was found to be optimal, with 40·5% of NEJ migrating after 150 min. This study describes a readily-reproducible method for studying the migration of F. hepatica NEJ within the definitive host. It will be useful for identifying potential drug and vaccine targets.

  12. A mucin-like peptide from Fasciola hepatica induces parasite-specific Th1-type cell immunity.

    PubMed

    Noya, Verónica; Brossard, Natalie; Berasaín, Patricia; Rodríguez, Ernesto; Chiale, Carolina; Mazal, Daniel; Carmona, Carlos; Freire, Teresa

    2016-03-01

    Fasciolosis, caused by the liver fluke Fasciola hepatica, is a major parasitic disease of livestock that causes significant economic losses worldwide. Although drugs are effective against liver flukes, they do not prevent reinfection, and continuous treatment is costly. Moreover, resistant fluke strains are emerging. In this context, vaccination is a good alternative since it provides a cost-effective long-term prevention strategy to control fasciolosis. In this paper, we evaluate the Fhmuc peptide as a potential vaccine against fasciolosis. This peptide derives from a mucin-like protein highly expressed in the infective stage of Fasciola hepatica. Mucin-like molecules expressed by parasites can contribute to several infection processes by protecting the parasite from host proteases and recognition by the immune system. We show that the Fhmuc peptide induces Th1-like immune responses specific for F. hepatica excretion-secretion products (FhESP) with a high production of IFNγ. We also investigated whether this peptide could protect animals from infection, and present preliminary data indicating that animals treated with Fhmuc exhibited reduced liver damage compared to non-immunised animals and that this protection was associated with a recruitment of B and T lymphocytes in the peritoneum, as well as eosinophils and mature dendritic cells. These results suggest that the mucin-like peptide Fhmuc could constitute a potential vaccine candidate against fasciolosis and pave the way towards the development of vaccines against parasites.

  13. Morphological and molecular characterization of lymnaeid snails and their potential role in transmission of Fasciola spp. in Vietnam.

    PubMed

    Dung, Bui Thi; Doanh, Pham Ngoc; The, Dang Tat; Loan, Ho Thi; Losson, Bertrand; Caron, Yannick

    2013-12-01

    Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.

  14. Time-Course Study of the Transcriptome of Peripheral Blood Mononuclear Cells (PBMCs) from Sheep Infected with Fasciola hepatica

    PubMed Central

    Scheerlinck, Jean-Pierre; Ansell, Brendan R. E.; Hall, Ross S.; Gasser, Robin B.; Jex, Aaron R.

    2016-01-01

    Fasciola hepatica is a parasitic trematode that infects a wide range of mammalian hosts, including livestock and humans, in temperate and tropical regions globally. This trematode causes the disease fascioliasis, which consists of an acute phase (≤ 12 weeks) during which juvenile parasites migrate through the host liver tissues, and a chronic phase (> 12 weeks) following the establishment of adult parasites in the liver bile ducts. Few studies have explored the progression of the host response over the course of Fasciola infection in the same animals. In this study, we characterized transcriptomic changes in peripheral blood mononuclear cells (PBMCs) collected from sheep at three time points over the first eight weeks of infection relative to uninfected controls. In total, 183 and 76 genes were found to be differentially transcribed at two and eight weeks post-infection respectively. Functional and pathway analysis of differentially transcribed genes revealed changes related to T-cell activation that may underpin a Th2-biased immune response against this parasite. This first insight into the dynamics of host responses during the early stages of infection improves the understanding of the pathogenesis of acute fascioliasis, informs vaccine development and presents a set of PBMC markers with diagnostic potential. PMID:27438474

  15. Genotypic characterization and species identification of Fasciola spp. with implications regarding the isolates infecting goats in Vietnam.

    PubMed

    Nguyen, Thanh Giang Thi; Van De, Nguyen; Vercruysse, Jozef; Dorny, Pierre; Le, Thanh Hoa

    2009-12-01

    Ribosomal RNA sequences (361 or 362bp) of the second internal transcribed spacer 2 (ITS-2) and a portion of mitochondrial cox1 (423bp) for Fasciola spp. obtained from specimens collected in indigenous and hybrid goats and sheep in Vietnam were characterized for genotypic status and hybridization/introgression. Alignment of 48 ITS-2 sequences (also those from goats and sheep in this study) indicates that F. gigantica and F. hepatica differ typically from each other at seven sites whereas one of these is a distinguishing deletion (T) at the 327th position in F. gigantica relative to F. hepatica. The isolates from the mountainous goats in the North of Vietnam (Yen Bai province) showed the ITS-2 composition relatively identical to that of F. hepatica. The ITS-2 sequences from populations of Fasciola isolates in goats had probably experienced introgression/hybridization as reported previously in other ruminants and humans. All Vietnamese goat-of-origin specimens had high pairwise percentage of mitochondrial cox1 sequences to F. gigantica (97-100%), and very low identity to F. hepatica (91-93%), suggesting their maternal linkage to be traced to F. gigantica. The presence of hybrid and/or introgressed populations of liver flukes bearing genetic material from both F. hepatica and F. gigantica in the goats/sheep in Vietnam, regardless of indigenous or imported hosts, appears to be the first demonstration from a tropical country. PMID:19733565

  16. Ultrastructure and development of Nosema podocotyloidis n. sp. (Microsporidia), a hyperparasite of Podocotyloides magnatestis (Trematoda), a parasite of Parapristipoma octolineatum (Teleostei)

    PubMed Central

    2014-01-01

    Nosema podocotyloidis n. sp. (Microsporidia, Nosematidae) is described from Podocotyloides magnatestis (Trematoda: Opecoelidae), a parasite of the fish Parapristipoma octolineatum (Teleostei) in the Atlantic Ocean. Electron microscopy reveals that all the stages of the cycle (merogony and sporogony) are diplokaryotic and in direct contact with the cytoplasm of host cells. There is no sporophorous vesicle (pansporoblast). The earliest stages observed are meronts, which have a simple plasmic membrane. Their cytoplasm is granular, rich in ribosomes and contains some sacculi of endoplasmic reticulum. They divide by binary fission into diplokaryotic sporonts. The sporonts have a thick electron-dense wall. Their diplokaryon is slightly less electron-dense than the cytoplasm. The cytoplasm of more advanced sporonts has numerous electron-lucent vesicles. Sporonts with two diplokarya divide by binary fission into diplokaryotic sporoblasts. The older sporoblasts are irregular or elongate and the polar filament is in formation. Their cytoplasm is denser, with ribosomes and lamellae of granular endoplasmic reticulum. The sporoblasts evolve into spores. The mature spores are broadly oval and measure 3.6 (3.1–4.0) × 2.58 (1.8–3.3) μm. Their wall is 100–300 nm thick. The polar tube is isofilar with 11–16 coils, 130–155 nm in diameter and arranged in many layers in the centre of the spore. The polaroplast is divided into two regions: an outer electron-dense cup with granular content and lacking lamellae and an internal region, less electron-dense, composed of irregularly arranged sacs. The posterior vacuole, with an amorphous electron-dense content, is present. The new species is compared with other species of Nosema from trematodes. PMID:25174849

  17. Ultrastructure and development of Nosema podocotyloidis n. sp. (Microsporidia), a hyperparasite of Podocotyloides magnatestis (Trematoda), a parasite of Parapristipoma octolineatum (Teleostei).

    PubMed

    Toguebaye, Bhen Sikina; Quilichini, Yann; Diagne, Papa Mbagnick; Marchand, Bernard

    2014-01-01

    Nosema podocotyloidis n. sp. (Microsporidia, Nosematidae) is described from Podocotyloides magnatestis (Trematoda: Opecoelidae), a parasite of the fish Parapristipoma octolineatum (Teleostei) in the Atlantic Ocean. Electron microscopy reveals that all the stages of the cycle (merogony and sporogony) are diplokaryotic and in direct contact with the cytoplasm of host cells. There is no sporophorous vesicle (pansporoblast). The earliest stages observed are meronts, which have a simple plasmic membrane. Their cytoplasm is granular, rich in ribosomes and contains some sacculi of endoplasmic reticulum. They divide by binary fission into diplokaryotic sporonts. The sporonts have a thick electron-dense wall. Their diplokaryon is slightly less electron-dense than the cytoplasm. The cytoplasm of more advanced sporonts has numerous electron-lucent vesicles. Sporonts with two diplokarya divide by binary fission into diplokaryotic sporoblasts. The older sporoblasts are irregular or elongate and the polar filament is in formation. Their cytoplasm is denser, with ribosomes and lamellae of granular endoplasmic reticulum. The sporoblasts evolve into spores. The mature spores are broadly oval and measure 3.6 (3.1-4.0) × 2.58 (1.8-3.3) μm. Their wall is 100-300 nm thick. The polar tube is isofilar with 11-16 coils, 130-155 nm in diameter and arranged in many layers in the centre of the spore. The polaroplast is divided into two regions: an outer electron-dense cup with granular content and lacking lamellae and an internal region, less electron-dense, composed of irregularly arranged sacs. The posterior vacuole, with an amorphous electron-dense content, is present. The new species is compared with other species of Nosema from trematodes. PMID:25174849

  18. Invertebrate host-parasite relationships: convergent evolution of a tropomyosin epitope between Schistosoma sp., Fasciola hepatica, and certain pulmonate snails.

    PubMed

    Weston, D; Allen, B; Thakur, A; LoVerde, P T; Kemp, W M

    1994-05-01

    Monoclonal antibodies (mAb) directed against Schistosoma mansoni tropomyosin isoform, SMTM (Xu et al. Experimental Parasitology 69, 373-392, 1989), were used to test for cross-reactivity with Biomphalaria glabrata antigens. One mAb (1F10) recognized antigens of 39, 41, and 80 kDa in a snail head/foot antigen preparation but not a hepatopancreas antigen preparation. Another mAb (1C1) cross-reacted with a 39-kDa antigen in the head/foot extract but not in the hepatopancreas extract. Epitope mapping revealed the 1F10 epitope to be between amino acids 135 and 188 of both Bg39 (Dissous et al. Molecular and Biochemical Parasitology 43, 245-256, 1990) and BgTMII (Weston and Kemp, Experimental Parasitology 76, 358-370, 1993), while the 1C1 epitope was located between amino acids 189 and 213 of BgTMII. Various invertebrate species, including members from Trematoda, Pulmonata, Annelida, and Arthropoda, were tested for cross-reactivity with the monoclonal antibodies. While the 1F10 mAb displayed broad invertebrate cross-reactivity, the 1C1 mAb cross-reactivity was restricted to schistosomes, F. hepatica, and the pulmonate snails B. glabrata and Physa sp. PMID:7512930

  19. Lymnaea palustris and Lymnaea fuscus are potential but uncommon intermediate hosts of Fasciola hepatica in Sweden

    PubMed Central

    2013-01-01

    Background Lymnaea palustris and L. fuscus are members of the European stagnicolines (Gastropoda: Lymnaeidae). The role of stagnicolines in transmission of Fasciola hepatica has been often proposed. To assess the possible relationship between these two stagnicolines and F. hepatica in Sweden, field monitoring in parallel with experimental infections of L. palustris and L. fuscus were conducted. Methods Stagnicoline snails were collected and identified on pastures grazed by either sheep or cattle on four farms suffering from fasciolosis in Sweden during 2011–2012. Field-collected L. palustris and L. fuscus were examined for F. hepatica DNA by PCR. In the laboratory, different age groups of L. palustris, L. fuscus and G. truncatula were each exposed to two F. hepatica miracidia and main infection characteristics were obtained. Results One field-collected L. palustris (out of n = 668) contained F. hepatica as determined by PCR. On the other hand, stagnicolines artificially exposed to F. hepatica miracidia resulted in successful infection with fully differentiated cercariae, but only in juvenile snails (size, 1–2 mm at exposure) and with a prevalence of 51% and 13% in L. palustris and L. fuscus, respectively. In contrast, 90% of juvenile (size, 1–2 mm) and 92% of preadult G. truncatula (size, ≥ 2-4 mm), respectively, were successfully infected. Delayed, reduced and/or no spontaneous cercarial shedding was observed in the two stagnicolines when compared to G. truncatula. However, at snail dissection most cercariae from L. fuscus and L. palustris were able to encyst similarly to those from G. truncatula. Conclusion Both L. fuscus and L. palustris can sustain larval development of F. hepatica but with an apparent level of age resistance. The finding of a single F. hepatica positive specimen of L. palustris, together with infection characteristics from the experimental infection, suggest that L. palustris is a more suitable snail vector of F. hepatica than L

  20. The effect of Fasciola hepatica infection on respiratory vaccine responsiveness in calves.

    PubMed

    Krump, L; Hamilton, C M; Sekiya, M; O'Neill, R; Mulcahy, G

    2014-03-17

    Fasciola hepatica is a common parasite in cattle, and bovine fasciolosis causes significant production losses, as well as being a zoonotic disease of global importance. F. hepatica has been shown to have immunoregulatory effects and the aim of this research was to establish whether F. hepatica infection influences the response to vaccination against respiratory pathogens in calves. A total of 48 calves were randomly and equally allocated to two groups. The experimental group was infected with F. hepatica, while the other group was used as a control. At week 2 and 6 after infection calves from both groups were administered a vaccine containing inactivated PI-3, BRSV and Mannheimia haemolytica, pathogens commonly associated with bovine respiratory disease. Blood samples were taken weekly over 12 weeks to measure specific antibodies against all vaccine antigens and against F. hepatica, as well as IgG1 and IgG2 isotypes for PI-3 and BRSV specific antibodies. Faecal samples were examined for F. hepatica eggs and routine haematology and liver enzyme biochemistry were performed and cytokine production in vitro measured. Liver enzymes (GGT and GLDH) and eosinophils were significantly higher in the experimental group, whereas neutrophil numbers were higher in the control group. There was no significant difference between the groups in terms of vaccine-specific total responses to PI-3, BRSV and M. haemolytica. IgG1 levels were higher in comparison to IgG2 levels in both PI-3 and BRSV specific antibodies. IL-4 levels from stimulated and unstimulated PBMC were significantly higher in the control group. IFN-γ levels were significantly higher in PBMC from the control group when cultured in medium only. No significant differences were noted in the levels of other cytokines measured. In this work, no effect of early F. hepatica infection on the antibody responses to a range of respiratory vaccine antigens in calves was shown. However, differences in cytokine responsiveness of

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

    PubMed

    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. PMID:24736227

  2. Towards assessing fine-scale indicators for the spatial transmission risk of Fasciola hepatica in cattle.

    PubMed

    Charlier, Johannes; Bennema, Sita Carolien; Caron, Yannick; Counotte, Michel; Ducheyne, Els; Hendrickx, Guy; Vercruysse, Jozef

    2011-05-01

    In order to improve the spatial resolution of current risk maps for fasciolosis in cattle, more knowledge is needed with respect to farm-level factors that determine infection risk. In this study, we visited 39 dairy farms within a predefined low- and high-risk area for fasciolosis in Belgium and assessed their infection status by an indirect bulk tank milk enzyme-linked immunosorbent assay (ELISA). Management factors were collected and all pastured lands of the farms were visited to identify and georeference potential snail habitats. The habitats were visually characterised, investigated for the presence of the intermediate host snails of Fasciola hepatica (i.e. Galba truncatula and Radix spp) and used in a geographical information system (GIS) to construct overlays including information on soil and hydrology. A linear regression model was used to evaluate associations between bulk tank milk ELISA results and farm level management and habitat factors. A logistic, mixed model was used to identify possible risk factors for the presence of intermediate host snails on a potential habitat. Potential snail habitats were found in 35 out of 39 farms. A total of 87 potential habitats were identified and on 29% of these, intermediate host snails were found. The number of potential habitats, the presence of snails, drainage of pastures, month of turnout of the cows, stocking rate, type of watering place and risk area were significantly associated with the bulk tank milk ELISA result and explained 85% of the observed variation. Intermediate host snails were more likely to be present with increasing surface of the potential habitat and on loamy soils. This study confirms the importance of farm management factors in the infection risk for F. hepatica in cattle and highlights that the combination of management factors with characterization of snail habitats is a powerful means to predict the infection risk with F. hepatica at the individual farm level. Further research is needed

  3. Fasciola gigantica: anthelmintic effect of the aqueous extract of Artocarpus lakoocha.

    PubMed

    Saowakon, Naruwan; Tansatit, Tawewan; Wanichanon, Chaitip; Chanakul, Waraporn; Reutrakul, Vichai; Sobhon, Prasert

    2009-08-01

    The effect of the crude extract of Artocarpus lakoocha (70% composition is 2,4,3',5'- tetrahydroxystilbene -THS) on adult Fasciola gigantica was evaluated after incubating the parasites in M-199 medium containing 250, 500, 750 and 1000 microg/ml of the crude extract, or triclabendazole (TCZ) at the concentrations of 80 and 175 microg/ml as the positive control, for 3, 6, 12 and 24h, using relative motility (RM) assay and observation by scanning electron microscope (SEM). Decreased contraction and motility were first observed after 3h incubation with TCZ at the concentration 80 and 175 microg/ml. TCZ markedly reduced the parasite's motility at the concentration of 175 microg/ml at 6h, and killed the worms after 12h exposure. The crude extract of A. lakoocha at all concentrations reduced the parasite's motility similar to TCZ at 3h incubation. In 250 and 500 microg/ml of the crude extract, the values were decreased from 3 to 12h, then they were stable between 12 and 24h and reduced to the level approximately 30-40% of the control. At 750 and 1000 microg/ml concentrations the crude extract rapidly reduced the RM values from the start to 12h and killed the parasites between 12 and 24h incubation. The crude extract also inhibited the larval migration by 75% and 100% at the concentrations of 250-500 and 750-1000 microg/ml, respectively. TCZ and the crude extract caused sequentially changes in the tegument including swelling, followed by blebbings that later ruptured, leading to the erosion and desquamation of the tegument syncytium. As the result, lesion was formed which exposed the basal lamina. The damage appeared more severe on the dorsal than the ventral surface, and earlier on the anterior part and lateral margins when compared to the posterior part. The severity and rapidity of the damages were enhanced with increasing concentration of the crude extract. Hence, the crude extract of A. lakoocha, may exert its fasciolicidal effect against adult F. gigantica by

  4. Variations in cercarial production and the level of in vitro activation of metacercariae of two different isolates of Fasciola hepatica.

    PubMed

    Januszkiewicz, Kamil; Norbury, Luke J; Wilkowski, Przemysław; Zawistowska-Deniziak, Anna; Wesołowska, Agnieszka; Wedrychowicz, Halina

    2015-09-01

    Fasciola hepatica infections cause large economic losses and are a serious veterinary medicine problem in many regions of the world. Recent studies examining fascioliasis in the bison population from Bialowieza National Park have shown that the prevalence of infection with this parasite is up to 100%. Liver flukes isolated from bison from Bialowieza National Park in Poland were compared with a fluke strain originally obtained from the Central Veterinary Laboratory, Weybridge, UK, to determine variations in cercarial production and establish the ability of their metacercariae to activate in vitro. Some small differences in cercarial production between the two isolates are shown, while significant differences in the ability of their metacercariae to activate in vitro were observed. PMID:26204191

  5. A Case of Probable Mixed-Infection with Clonorchis sinensis and Fasciola sp.: CT and Parasitological Findings

    PubMed Central

    Kim, Tae Yun; Lee, Yun-Sik; Yun, Ji Hye; Kim, Jeong Ju; Choi, Won Hyung; Oh, In Hwan; Song, Hyun Ouk

    2010-01-01

    We report here a human case probably mixed-infected with Clonorchis sinensis and Fasciola sp. who was diagnosed by computed tomography (CT) scan, serological findings, and/or fecal examination. The patient was a 43-year-old Korean female and was admitted to Kyung Hee University Hospital with the complaints of fever and abdominal pain. On admission, marked eosinophilia was noted in her peripheral blood. CT scan showed specific lesions for clonorchiasis and fascioliasis in the liver, along with lesions suggestive of amebic abscess. Micro-ELISA revealed positive results for the 2 helminthic infections. Eggs of C. sinensis and trophozoites of Entamoeba histolytica were observed in the stool. Treatment with praziquantel followed by metronidazole and tinidazole reduced abnormalities in the liver and eosinophilia. This is the first case report of a possible co-infection with 2 kinds of liver flukes in the Republic of Korea. PMID:20585533

  6. Physiological responses of insular wild black rat (Rattus rattus) to natural infection by the digenean trematode Fasciola hepatica.

    PubMed

    Magnanou, E; Fons, R; Feliu, C; Morand, S

    2006-06-01

    Wild black rat Rattus rattus is regularly infected by the liver fluke Fasciola hepatica on Corsica. This report constitutes the only example of a murid rodent that plays an important epidemiological role for the Fasciolosis. We investigated the influence of such unusual parasite infection on black rat physiology by measuring its oxygen consumption at different ambient temperatures. Black rat energy requirements are influenced by body mass, temperature of the experiment and parasite infestation. The influence of the presence of F. hepatica was more pronounced for cold temperatures. The mean increase of 56% in oxygen requirements for infected rats is extremely high, indeed unexpected, according to previous knowledge. These high physiological constraints may be explained by the recent confrontation of the digenean and the rodent.

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

    PubMed Central

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

    2015-01-01

    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]. PMID:26697343

  8. Increase of gluthatione S-transferase, carboxyl esterase and carbonyl reductase in Fasciola hepatica recovered from triclabendazole treated sheep.

    PubMed

    Scarcella, S; Solana, M V; Fernandez, V; Lamenza, P; Ceballos, L; Solana, H

    2013-10-01

    Fasciolasis is a zoonotic parasitic disease caused by Fasciola hepatica and its control is mainly based on the use of triclabendazole (TCBZ). Parasite resistance to different anthelmintics is growing worldwide, including the resistance of F. hepatica to TCBZ. In the present work we evaluate "in vivo" the activity of xenobiotic metabolizing enzymes of phase I (carboxyl esterases) and phase II (glutathione S-transferases and carbonyl reductases) recovered of flukes from sheep treated with TCBZ. All three enzymes showed increased activity in TCBZ flukes returning 60h post-treatment at similar to baseline unexposed flukes. TCBZ action may induce secondary oxidative stress, which may explain the observed increment in activities of the analyzed enzymes as a defensive mechanism. The enzymes analyzed are candidates to participate actively in the development of resistance at TCBZ in F. hepatica.

  9. Cloning and characterization of a female genital complex cDNA from the liver fluke Fasciola hepatica.

    PubMed Central

    Zurita, M; Bieber, D; Ringold, G; Mansour, T E

    1987-01-01

    A cDNA clone whose RNA is abundant in the female genital complex of the liver fluke Fasciola hepatica has been isolated from a cDNA library in lambda gt10 by differential screening. The pattern of expression in different fluke tissues and at different stages of miracidium formation suggests that this gene is expressed in the F. hepatica vitelleria. The nucleotide sequence of the cloned cDNA was determined and the primary structure of the putative protein was deduced. The proposed protein is rich in glycine, lysine, and tyrosine and its overall amino acid composition agrees with that reported for the F. hepatica egg shell. The clone has homology with DNA from other trematodes; this homology is higher in organisms in which egg development is similar to that of F. hepatica and suggests that the protein is conserved in organisms in which miracidium formation occurs in fresh water. Images PMID:3470798

  10. Seasonal growth and mortality of juveniles of Lampsilis fasciola (Bivalvia: Unionidae) released to a fish hatchery raceway

    USGS Publications Warehouse

    Hanlon, S.D.; Neves, R.J.

    2006-01-01

    Recent efforts to restore remnant or extirpated populations of freshwater mussels have focused on artificial propagation as an effective and practical conservation strategy. Although artificially cultured juveniles have been produced and released to the wild at various times of the year, no study has investigated the best time of year to release these juveniles. Newly metamorphosed juveniles of the wavyrayed lampmussel (Lampsilis fasciola) were released into a stream-fed fish hatchery raceway during March, June, and September. Growth and survival rates were measured 32, 52, 72, and 92 days post-metamorphosis. Juveniles released in June experienced the greatest growth and survival rates. Juveniles released in September and March experienced high mortality within the first month of release and exhibited poor growth in the cool water conditions typical of those seasons. Overwinter survival exhibited a size-dependent relationship.

  11. Pseudosuccinea columella (Say 1817) (Gastropoda, Lymnaeidae), snail host of Fasciola hepatica: first record for France in the wild.

    PubMed

    Pointier, Jean-Pierre; Coustau, Christine; Rondelaud, Daniel; Theron, André

    2007-10-01

    The lymnaeid snail Pseudosuccinea columella has shown strong invasive capabilities in the last decades, and this species has now a worldwide distribution. So far, the presence of this snail in Europe was restricted to botanical gardens, but the recent discovery of a few specimens along the banks of the Lot River, southwestern France is the first record of this species in the wild. The first generation of this P. columella isolate obtained in the laboratory was used for parasitological tests with a French Fasciola hepatica sample. Experimental infections showed that 100% of snails (n=26) developed infection when exposed to F. hepatica miracidia. Consequences for the epidemiology of F. hepatica transmission in France are discussed.

  12. Validation of an interactive map assessing the potential spread of Galba truncatula as intermediate host of Fasciola hepatica in Switzerland.

    PubMed

    Baggenstos, Rhea; Dahinden, Tobias; Torgerson, Paul R; Bär, Hansruedi; Rapsch, Christina; Knubben-Schweizer, Gabriela

    2016-01-01

    Bovine fasciolosis, caused by Fasciola hepatica, is widespread in Switzerland. The risk regions were modelled in 2008 by an interactive map, showing the monthly potential risk of transmission of F. hepatica in Switzerland. As this map is based on a mathematical model, the aim of the present study was to evaluate the interactive map by means of a field survey taking different data sources into account. It was found that the interactive map has a sensitivity of 40.7-88.9%, a specificity of 11.4-18.8%, a positive predictive value of 26.7-51.4%, and a negative predictive value of 13.1-83.6%, depending on the source of the data. In conclusion, the grid of the interactive map (100 x 100 m) does not reflect enough detail and the underlying model of the interactive map is lacking transmission data. PMID:27245800

  13. The combined effect of isolation and Fasciola hepatica infection on the life history traits of Fossaria cubensis.

    PubMed

    Gutierrez, A; Yong, M; Wong, L; Sánchez, J

    2001-08-01

    Life history traits of Fossaria cubensis were compared between isolated and paired snails after infection with three miracidia of Fasciola hepatica. Four experimental groups were tested: isolated-unexposed, paired-unexposed, isolated-infected, and paired-infected. A repeated-measures ANOVA showed statistically significant interactions among isolation, infection, and age effects for shell size, number of egg masses per snail, number of eggs per snail, and number of viable eggs per snail. Isolated-unexposed snails exhibited the higher values of these variables and those of survival and finite and intrinsic rates of natural increase. Infection stimulated shell growth during the prepatent period, but differences were present only in paired snails since isolation causes a similar effect. Reproduction, in terms of the number of egg masses per snail and the number of eggs per mass per snail, decreases in the presence of parasitic infection, whereas isolation stimulates it. These effects were observed from early stages of infection.

  14. Variations in cercarial production and the level of in vitro activation of metacercariae of two different isolates of Fasciola hepatica.

    PubMed

    Januszkiewicz, Kamil; Norbury, Luke J; Wilkowski, Przemysław; Zawistowska-Deniziak, Anna; Wesołowska, Agnieszka; Wedrychowicz, Halina

    2015-09-01

    Fasciola hepatica infections cause large economic losses and are a serious veterinary medicine problem in many regions of the world. Recent studies examining fascioliasis in the bison population from Bialowieza National Park have shown that the prevalence of infection with this parasite is up to 100%. Liver flukes isolated from bison from Bialowieza National Park in Poland were compared with a fluke strain originally obtained from the Central Veterinary Laboratory, Weybridge, UK, to determine variations in cercarial production and establish the ability of their metacercariae to activate in vitro. Some small differences in cercarial production between the two isolates are shown, while significant differences in the ability of their metacercariae to activate in vitro were observed.

  15. Comparison of Fasciola hepatica genotypes in relation to their ability to establish patent infections in the final host.

    PubMed

    Zintl, Annetta; Talavera, Silvia; Sacchi-Nestor, Carlotta; Ryan, Marion; Chryssafidis, Andreas; Mulcahy, Grace

    2015-06-15

    Fasciola hepatica is a common and economically important parasite of sheep and cattle. Although its marked genetic heterogeneity is well recognised, an association between haplotypes and specific phenotypic traits has yet to be identified. Using experimental infections in cattle this study investigated whether a fragment of mitochondrial DNA (coding for cytochrome c oxidase subunit III, transfer RNA histidine and cytochrome b) and 3 nuclear microsatellite loci (Fh15, Fh23 and Fh25) could be used as markers for the parasite's ability to complete its tissue migration and establish in the liver of the final host. While we did not detect any shift in the frequency of the various genotypes in the population of metacercariae used for the infection on the one hand and the flukes collected from the liver on the other, there was an indication that parasites with heterozygous microsatellite alleles may have a selective advantage over homozygote parasites during their migration in the final host.

  16. Fasciola hepatica in Cuba: compatibility of different isolates with two intermediate snail hosts, Galba cubensis and Pseudosuccinea columella.

    PubMed

    Vázquez, A A; Sánchez, J; Pointier, J-P; Théron, A; Hurtrez-Boussès, S

    2014-12-01

    In Cuba, only two lymnaeid snails, Galba cubensis and Pseudosuccinea columella, with different ecology and distribution patterns, are intermediate hosts for Fasciola hepatica. The compatibility of these two species as hosts was analysed through their rates of infection, the production of rediae and survivorship when exposed to F. hepatica miracidia. Ten populations of G. cubensis, eight of P. columella collected from various habitats and six isolates of F. hepatica sampled in slaughterhouses from different localities were tested. Our results clearly demonstrate that G. cubensis is a more compatible host for F. hepatica in Cuba when compared with P. columella. However, the role that P. columella may have in fascioliasis transmission under certain conditions should not be disregarded. Variation in infectivity among isolates of F. hepatica were also observed and may explain why some regions in Cuba are more commonly subjected to fascioliasis outbreaks.

  17. Presence of very high prevalence and intensity of infection with Fasciola hepatica among Aymara children from the Northern Bolivian Altiplano.

    PubMed

    Esteban, J G; Flores, A; Aguirre, C; Strauss, W; Angles, R; Mas-Coma, S

    1997-06-24

    Coprological studies of school children from four communities in the Northern Bolivian Altiplano were carried out in order to estimate the prevalences and intensities of Fasciola hepatica infection. Single stool specimens were collected at random from 558 school children (308 boys and 250 girls) aged 5-19 years old. Nineteen different parasite species (13 protozoan and six helminths) were detected. Of the children examined, 98.7% (96.5-100%) presented infection with at least one parasite species. The mean prevalence of 27.6% by Fasciola hepatica (range, 5.9-38.2%) was the highest not only with respect to the helminth species found in the Northern Bolivian Altiplano but also among the fasciolosis prevalences reported in children in other parts of the world to date. Prevalences were significantly different among the communities surveyed and was significantly higher in the 9-12 years age group. There were, however, no significant differences between sexes. Among the 154 children presenting F. hepatica eggs in stools, intensities ranged from 24-5064 eggs per gram of faeces (epg), with arithmetic and geometric means of 474 and 201 epg, respectively. Significant differences in mean egg output were detected between communities, sexes and age groups. Individual fasciolosis infections coexisting with other pathogenic parasite species (Entamoeba histolytica and/or E. dispar, Giardia intestinalis, Balantidium coli, Dientamoeba fragilis, Cryptosporidium sp., Hymenolepis nana, Taenia spp., Trichuris trichiura, Ascaris lumbricoides and Enterobius vermicularis) were detected. A significant positive association with F. hepatica was only found in the case of G. intestinalis. This coprological study not only verifies the existence of high prevalences of F. hepatica among humans in the Northern Bolivian Altiplano, but also demonstrates the need to expand the Southern boundaries of this high endemic zone to include the Southeastern region of Lake Titicaca.

  18. Ovicidal effect of the methanolic extract of ginger (Zingiber officinale) on Fasciola hepatica eggs: an in vitro study.

    PubMed

    Moazeni, Mohammad; Khademolhoseini, Ali Asghar

    2016-09-01

    Fasciolosis is of considerable economic and public health importance worldwide. Little information is available on the ovicidal effects of anthelminthic drugs. The use of ovicidal anthelmintics can be effective in disease control. In this study, the effectiveness of the methanolic extract of ginger (Zingiber officinale) on the eggs of Fasciola hepatica is investigated. Fasciola hepatica eggs were obtained from the gall bladders of naturally infected sheep and kept at 4 °C until use. The eggs were exposed to varying concentrations of ginger extract (1, 5, 10, 25 and 50 mg/mL) for 24, 48 and 72 h. To investigate the effect of the ginger extracts on the miracidial formation, the treated eggs were incubated at 28 °C for 14 days. The results indicated that F. hepatica eggs are susceptible to the methanolic extract of Z. officinale. The ovicidal effect of ginger extract at a concentration of 1 mg/mL with 24, 48 and 72 h treatment time was 46.08, 51.53 and 69.09 % respectively (compared with 22.70 % for control group). The ovicidal effect of ginger extract at a concentration of 5 mg/mL after 24 h was 98.84 %. One hundred percent ovicidal efficacy was obtained through application of ginger extract at concentrations of 5 and 10 mg/mL with a 48 and 24 h treatment time respectively. The in vitro ovicidal effect of the methanolic extract of Z. officinale was satisfactory in this study, however, in vivo efficacy of this extract, remains for further investigation. To the best of our knowledge, this is the first report on the ovicidal effect of Z. officinale against F. hepatica eggs.

  19. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro.

    PubMed

    McCusker, Paul; McVeigh, Paul; Rathinasamy, Vignesh; Toet, Hayley; McCammick, Erin; O'Connor, Anna; Marks, Nikki J; Mousley, Angela; Brennan, Gerard P; Halton, David W; Spithill, Terry W; Maule, Aaron G

    2016-09-01

    Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving 'molecular toolbox' for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the "neoblast" stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in vitro study

  20. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro.

    PubMed

    McCusker, Paul; McVeigh, Paul; Rathinasamy, Vignesh; Toet, Hayley; McCammick, Erin; O'Connor, Anna; Marks, Nikki J; Mousley, Angela; Brennan, Gerard P; Halton, David W; Spithill, Terry W; Maule, Aaron G

    2016-09-01

    Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving 'molecular toolbox' for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the "neoblast" stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in vitro study

  1. Presence of very high prevalence and intensity of infection with Fasciola hepatica among Aymara children from the Northern Bolivian Altiplano.

    PubMed

    Esteban, J G; Flores, A; Aguirre, C; Strauss, W; Angles, R; Mas-Coma, S

    1997-06-24

    Coprological studies of school children from four communities in the Northern Bolivian Altiplano were carried out in order to estimate the prevalences and intensities of Fasciola hepatica infection. Single stool specimens were collected at random from 558 school children (308 boys and 250 girls) aged 5-19 years old. Nineteen different parasite species (13 protozoan and six helminths) were detected. Of the children examined, 98.7% (96.5-100%) presented infection with at least one parasite species. The mean prevalence of 27.6% by Fasciola hepatica (range, 5.9-38.2%) was the highest not only with respect to the helminth species found in the Northern Bolivian Altiplano but also among the fasciolosis prevalences reported in children in other parts of the world to date. Prevalences were significantly different among the communities surveyed and was significantly higher in the 9-12 years age group. There were, however, no significant differences between sexes. Among the 154 children presenting F. hepatica eggs in stools, intensities ranged from 24-5064 eggs per gram of faeces (epg), with arithmetic and geometric means of 474 and 201 epg, respectively. Significant differences in mean egg output were detected between communities, sexes and age groups. Individual fasciolosis infections coexisting with other pathogenic parasite species (Entamoeba histolytica and/or E. dispar, Giardia intestinalis, Balantidium coli, Dientamoeba fragilis, Cryptosporidium sp., Hymenolepis nana, Taenia spp., Trichuris trichiura, Ascaris lumbricoides and Enterobius vermicularis) were detected. A significant positive association with F. hepatica was only found in the case of G. intestinalis. This coprological study not only verifies the existence of high prevalences of F. hepatica among humans in the Northern Bolivian Altiplano, but also demonstrates the need to expand the Southern boundaries of this high endemic zone to include the Southeastern region of Lake Titicaca. PMID:9177091

  2. Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines.

    PubMed

    Dalton, John P; Neill, Sandra O; Stack, Colin; Collins, Peter; Walshe, Alan; Sekiya, Mary; Doyle, Sean; Mulcahy, Grace; Hoyle, Deborah; Khaznadji, Eric; Moiré, Nathalie; Brennan, Gerard; Mousley, Angela; Kreshchenko, Natalia; Maule, Aaron G; Donnelly, Sheila M

    2003-09-30

    Fasciola hepatica secretes cathepsin L proteases that facilitate the penetration of the parasite through the tissues of its host, and also participate in functions such as feeding and immune evasion. The major proteases, cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) are members of a lineage that gave rise to the human cathepsin Ls, Ks and Ss, but while they exhibit similarities in their substrate specificities to these enzymes they differ in having a wider pH range for activity and an enhanced stability at neutral pH. There are presently 13 Fasciola cathepsin L cDNAs deposited in the public databases representing a gene family of at least seven distinct members, although the temporal and spatial expression of each of these members in the developmental stage of F. hepatica remains unclear. Immunolocalisation and in situ hybridisation studies, using antibody and DNA probes, respectively, show that the vast majority of cathepsin L gene expression is carried out in the epithelial cells lining the parasite gut. Within these cells the enzyme is packaged into secretory vesicles that release their contents into the gut lumen for the purpose of degrading ingested host tissue and blood. Liver flukes also express a novel multi-domain cystatin that may be involved in the regulation of cathepsin L activity. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 have shown that these enzymes can induce protection, ranging from 33 to 79%, to experimental challenge with metacercariae of F. hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission. In this article we review the vaccine trials carried out over the past 8 years, the role of antibody and T cell responses in mediating protection and discuss the prospects of the cathepsin Ls in the development of first generation recombinant liver fluke vaccines.

  3. Ovicidal effect of the methanolic extract of ginger (Zingiber officinale) on Fasciola hepatica eggs: an in vitro study.

    PubMed

    Moazeni, Mohammad; Khademolhoseini, Ali Asghar

    2016-09-01

    Fasciolosis is of considerable economic and public health importance worldwide. Little information is available on the ovicidal effects of anthelminthic drugs. The use of ovicidal anthelmintics can be effective in disease control. In this study, the effectiveness of the methanolic extract of ginger (Zingiber officinale) on the eggs of Fasciola hepatica is investigated. Fasciola hepatica eggs were obtained from the gall bladders of naturally infected sheep and kept at 4 °C until use. The eggs were exposed to varying concentrations of ginger extract (1, 5, 10, 25 and 50 mg/mL) for 24, 48 and 72 h. To investigate the effect of the ginger extracts on the miracidial formation, the treated eggs were incubated at 28 °C for 14 days. The results indicated that F. hepatica eggs are susceptible to the methanolic extract of Z. officinale. The ovicidal effect of ginger extract at a concentration of 1 mg/mL with 24, 48 and 72 h treatment time was 46.08, 51.53 and 69.09 % respectively (compared with 22.70 % for control group). The ovicidal effect of ginger extract at a concentration of 5 mg/mL after 24 h was 98.84 %. One hundred percent ovicidal efficacy was obtained through application of ginger extract at concentrations of 5 and 10 mg/mL with a 48 and 24 h treatment time respectively. The in vitro ovicidal effect of the methanolic extract of Z. officinale was satisfactory in this study, however, in vivo efficacy of this extract, remains for further investigation. To the best of our knowledge, this is the first report on the ovicidal effect of Z. officinale against F. hepatica eggs. PMID:27605763

  4. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro

    PubMed Central

    Rathinasamy, Vignesh; Toet, Hayley; McCammick, Erin; O’Connor, Anna; Marks, Nikki J.; Mousley, Angela; Brennan, Gerard P.; Halton, David W.; Spithill, Terry W.; Maule, Aaron G.

    2016-01-01

    Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving ‘molecular toolbox’ for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the “neoblast” stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in

  5. Praziquantel efficacy against Brachylaima sp. metacercariae (Trematoda: Brachylaimidae) parasitizing the edible landsnail Cornu aspersum and its HPLC-MS/MS residue determination.

    PubMed

    Gállego, Laia; Gracenea, Mercedes

    2015-10-01

    Cornu aspersum is a terrestrial edible snail, often parasitized by Brachylaima (Trematoda) metacercariae. Ingestion of undercooked snails by humans allows metacercariae to develop to adult in the intestine causing brachylaimiasis (expected mortality rate 5-10%). The cosmopolitan character of the trematode and of its vector, enhanced in a future climate change scenario, and the absence of adequate sanitary controls of the snails in marketplaces clearly increase the risk of human brachylaimiasis. The treatment of farmed snails with praziquatel (PZQ) would be a tool to control this food-borne disease. The objectives of this study were: to report the prevalence of Brachylaima metacercariae parasitizing C. aspersum in marketplaces, to assess the efficacy and tolerance of PZQ in C. aspersum and to develop an HPLC-MS/MS analytical method to quantify PZQ residue in the edible snail (the acceptable daily intake of PZQ is 0.17 mg/kg bw in humans). Prevalence of parasitization of C. aspersum by Brachylaima metacercariae in public marketplaces reached the 80%. PZQ was orally administered to snails, mixed with the usual snail food. In dose determination assays three doses were individually tested (10 days): 1.2 mg PZQ/snail, 1.8 mg PZQ/snail (efficacy 97.3%, p < 0.001) and 2.4 mg PZQ/snail (efficacy 98.7%, p < 0.001). In dose confirmation tests (n = 200) the 2.4 mg PZQ/snail dose was group tested (10 snails/box, 7 days): 2 g of PZQ supplemented snail food were disposed daily in the treatment group boxes and 2 g of snail food (placebo) in the control boxes (efficacy 94.6%, p < 0.001; prevalence dropped from 68.7% in control group to 10.1% in treatment group, p < 0.001). Received dose was 220.2 mg PZQ/kg snail with shell. In the analytical method, linearity, lower limit of quantification (0.05 μg/ml), selectivity, carry over, accuracy, precision, dilution integrity, matrix effect and stability were tested. Sixty snails were treated (11 mg PZQ/g snail food

  6. Genetic Characterization of Fasciola Isolates from West Azerbaijan Province Iran Based on ITS1 and ITS2 Sequence of Ribosomal DNA

    PubMed Central

    GALAVANI, Hossein; GHOLIZADEH, Saber; HAZRATI TAPPEH, Khosrow

    2016-01-01

    Background: Fascioliasis, caused by Fasciola hepatica and F. gigantica, has medical and economic importance in the world. Molecular approaches comparing traditional methods using for identification and characterization of Fasciola spp. are precise and reliable. The aims of current study were molecular characterization of Fasciola spp. in West Azerbaijan Province, Iran and then comparative analysis of them using GenBank sequences. Methods: A total number of 580 isolates were collected from different hosts in five cities of West Azerbaijan Province, in 2014 from 90 slaughtered cattle (n=50) and sheep (n=40). After morphological identification and DNA extraction, designing specific primer were used to amplification of ITS1, 5.8s and ITS2 regions, 50 samples were conducted to sequence, randomly. Result: Using morphometric characters 99.14% and 0.86% of isolates identified as F. hepatica and F. gigantica, respectively. PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions. Sequence comparison among current study sequences and GenBank data showed 98% identity with 11 nucleotide mismatches. However, in phylogenetic tree F. hepatica sequences of West Azerbaijan Province, Iran, were in a close relationship with Iranian, Asian, and African isolates. Conclusions: Only F. hepatica species is distributed among sheep and cattle in West Azerbaijan Province Iran. However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp. Therefore, more studies are essential for designing new molecular markers to correct species identification. PMID:27095969

  7. Identification of a 17-kilodalton Fasciola hepatica immunodiagnostic antigen by the enzyme-linked immunoelectrotransfer blot technique.

    PubMed Central

    Hillyer, G V; Soler de Galanes, M

    1988-01-01

    Sera obtained from human patients, calves, sheep, and rabbits infected with Fasciola hepatica were tested by the Falcon assay screening test enzyme-linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immunoelectrotransfer blot (EITB) techniques with Fasciola hepatica excretory-secretory antigens in order to evaluate their immunodiagnostic potential. The study included sera from 13 patients infected with F. hepatica or a history suggesting fascioliasis, 5 patients infected and treated with bithionol or praziquantel (3 were cured with bithionol), 10 patients infected with Schistosoma mansoni, 6 infected with Trichinella spiralis, and 13 controls and sera from calves, sheep, and rabbits with a primary F. hepatica infection. By FAST-ELISA with F. hepatica excretory-secretory antigens, the serum samples from fascioliasis patients gave the highest absorbance values, and the schistosomiasis patient sera gave intermediate values compared with a normal human serum control. Also by FAST-ELISA, the values for serum from patients with fascioliasis decreased steadily after cure, reaching normal levels 20 to 47 weeks postcure. In contrast, the serum from two patients who had been treated but were not yet cured had high levels of antibodies for up to 3 years of infection. By EITB, the serum samples from humans, rabbits, cattle, and sheep with fascioliasis recognized two antigenic polypeptides of 17 and 63 kilodaltons (kDa) in the form of sharp bands. For humans, this recognition lasted for at least 3 years of infection. Sera from individuals with schistosomiasis mansoni or trichinosis or from normal controls did not recognize the 17-kDa F. hepatica antigenic polypeptide. However, serum from one human with S. mansoni and one with T. spiralis infection has slight bands in the 63-kDa region, suggesting cross-reactivity. Reactivity to the 17-kDa polypeptide was absent in fascioliasis patients at 1 year postcure. Reactivity to the 63-kDa polypeptide was significantly

  8. Early onset of changes to the reproductive system of Fasciola hepatica following in vivo treatment with triclabendazole.

    PubMed

    Hanna, R E B; Scarcella, S; Solana, H; McConnell, S; Fairweather, I

    2012-03-23

    Lambs infected with the Cullompton triclabendazole (TCBZ)-susceptible isolate of Fasciola hepatica were treated with TCBZ at a dosage of 10mg/kg at 16 weeks post-infection. Adult flukes were recovered from the liver at 3h, 24h, 48 h and 60 h post-treatment (pt). They were processed for histological analysis of the uterus, Mehlis' gland, vitellaria, ovary and testis. At 3h pt, the flukes were essentially similar to the controls and were producing normal eggs. Egg production had ceased by 24h pt. At this time period, the cells of the Mehlis' gland showed some evidence of vacuolation, but otherwise were relatively normal. A shift in the population of vitelline cells towards mature cells was observed at 24h pt, and this trend continued at later time-periods. It was accompanied by a breakdown of the cells and the presence of apoptotic bodies. Marked changes to the ovary were first noted at 48 h pt, as evidenced by vacuolation and the presence of apoptotic bodies. Some disruption to the testis was seen at 24h pt, with a reduction in the population of spermatogenic cells, the appearance of apoptotic bodies and some peripheral vacuolation of the tubules. These abnormalities increased in severity with longer time periods pt. The results bring forward the time-line of cessation of egg production by 24h, demonstrating that this process is affected very rapidly pt.

  9. Seroprevalence and GIS-supported risk factor analysis of Fasciola hepatica infections in dairy herds in Germany.

    PubMed

    Kuerpick, Birte; Conraths, Franz J; Staubach, Christoph; Fröhlich, Andreas; Schnieder, Thomas; Strube, Christina

    2013-07-01

    A total of 20 749 bulk tank milk (BTM) samples was collected in November 2008 from all over Germany, corresponding to 20.9% of all German dairy herds. The BTM samples were analysed for antibodies against Fasciola hepatica using the excretory-secretory (ES) ELISA. A geospatial map was drawn to show herd prevalences per postal code area. Various spatial risk factors were tested for potential statistical associations with the ELISA results in logistic regression supported by a geographical information system (GIS). The mean seroprevalence was 23.6% and prevalences in different German federal states varied between 2.6% and 38.4%. GIS analysis revealed statistically significant positive associations between the proportion of grassed area and water bodies per postal code area and positive BTM ELISA results. This can be explained by the biology of the intermediate host, the amphibious snail Galba (Lymnea) truncatula and the pasture-borne nature of fasciolosis. The full logistic regression model had a Pseudo-R 2 of 22%, while the final model obtained by controlled stepwise model building revealed a Pseudo-R 2 of 14%, indicating that additional, unrecorded factors and random effects contributed substantially to the occurrence of positive ELISA results. Considering the high seroprevalences in some areas and the economic impact of fasciolosis, farmers and veterinarians should be strongly advised to implement effective liver fluke control programmes.

  10. Fasciola hepatica: the infectivity of cattle-origin miracidia had increased over the past years in central France.

    PubMed

    Dreyfuss, G; Vignoles, P; Rondelaud, D

    2007-09-01

    A retrospective study on experimental infections of Galba truncatula, originating from four populations, was carried out to determine the infectivity of Fasciola hepatica miracidia in snails either subjected to single-miracidium or bimiracidial exposures in 2006 and to compare it with results found before 1990 in other experiments using the same protocol. In single-miracidium infections, the prevalence of snail infection was significantly greater in 2006 than in experiments performed before 1980, while snail survival at day 30 postexposure showed insignificant differences. In bimiracidial infections, snail survival in two snail groups was significantly lower in 2006 than in experiments performed from 1981 to 1990, while insignificant differences were found for the other two populations of G. truncatula. Compared to results found between 1981 and 1990, the prevalence of snail infection did not significantly differ in 2006. In each population considered alone, the mean number of cercariae shed by infected snails did not significantly vary, whatever the date of experiment and the number of miracidia used for each exposure. The greater infectivity of F. hepatica miracidia towards snails might be the consequence of specific molecules such as triclabendazole used since 1990 to treat fasciolosis in cattle from central France. PMID:17522892

  11. Fasciola hepatica: variations in redial development and cercarial production in relation to the geographic origin of the parasite.

    PubMed

    Rondelaud, Daniel; Sanabria, Rodrigo; Vignoles, Philippe; Dreyfuss, Gilles; Romero, Jorge

    2013-01-01

    Two hundred Galba truncatula, originating from a French population, were subjected to single-miracidium infections with an allopatric isolate (origin, Argentina) of Fasciola hepatica. The control group was constituted by 100 snails coming from the same population and exposed to sympatric miracidia of F. hepatica according to the same protocol. Snail samples were collected bimonthly from both groups between day 14 and day 112 p.e. (at 20 °C) and snail dissections were performed to count free rediae, intraredial morulae and free cercariae. Third and fourth generation rediae were significantly more numerous in the allopatric group, while the number of first generation rediae was significantly lower. In the sympatric group, the decrease in the number of intraredial morulae in the first, second and third redial generations was significantly faster. Free cercariae within the snail body were significantly more numerous in the sympatric than in the allopatric groups, whatever the date of snail dissection. The changes in redial development and cercarial production noted in the Argentinean group might be due to the evolution of South American flukes in a divergent way after the introduction of foreign infected ruminants in this continent from the 15th century. PMID:24054407

  12. MOLLUSCICIDAL AND LARVICIDAL ACTIVITIES OF Atriplex inflata AERIAL PARTS AGAINST THE MOLLUSK Galba truncatula, INTERMEDIATE HOST OF Fasciola hepatica.

    PubMed

    Hamed, Néjia; Njeh, Fatma; Damak, Mohamed; Ayadi, Ali; Mezghani-Jarraya, Raoudha; Hammami, Hayet

    2015-12-01

    Fasciolosis is a widespread parasitosis of farm live-stock in many developing countries. For this reason, it is necessary to search for new substances against parasitic diseases caused by flukes. Indeed, a wide variety of terrestrial plants have been subjected to chemical and pharmacological screening in order to discover their potential for human medicinal use. The molluscicidal and larvicidal activities of Atriplex inflata were tested on Galba truncatula and Fasciola hepatica larval stages infecting this snail in Tunisia. Phytochemical tests were conducted on extracts in order to establish a meaningful relationship with molluscicidal and larvicidal activities. The molluscicidal activity was evaluated by subjecting snails to sample aqueous solutions. Accordingly, hexane, ethyl acetate, methanol and methanol-water (8:2, v-v) were used as extraction solvents. As a result, hexane and ethyl acetate extracts showed potent activity, according to the World Health Organization, giving LC50 = 7.59 mg/L and 6.69 mg/L for hexane extracts of leaves and fruits, respectively. Ethyl acetate extracts gave LC50 = 5.90 mg/L and 7.32 mg/L for leaves and fruits, successively. Molluscicidal activities of powders were less potent on snails, but active according to the World Health Organization. Hexane and ethyl acetate extracts from leaves and fruits gave potent larvicidal activities with a delay rate exceeding 45.50% (45.50- 98.92%). Phytochemical tests showed that these activities may be attributed to the presence of triterpenoids and/or sterols.

  13. Liver fluke (Fasciola hepatica) naturally infecting introduced European brown hare (Lepus europaeus) in northern Patagonia: phenotype, prevalence and potential risk.

    PubMed

    Cuervo, Pablo F; Cataldo, Sophia Di; Fantozzi, M Cecilia; Deis, Erika; Isenrath, Gabriela Diaz; Viberti, Gabriela; Artigas, Patricio; Peixoto, Raquel; Valero, M Adela; Sierra, Roberto Mera Y; Mas-Coma, Santiago

    2015-09-01

    Fascioliasis has recently been included in the WHO list of Neglected Zoonotic Diseases. Besides being a major veterinary health problem, fascioliasis has large underdeveloping effects on the human communities affected. Though scarcely considered in fascioliasis epidemiology, it is well recognized that both native and introduced wildlife species may play a significant role as reservoirs of the disease. The objectives are to study the morphological characteristics of Fasciola hepatica adults and eggs in a population of Lepus europaeus, to assess liver fluke prevalence, and to analyze the potential reservoir role of the European brown hare in northern Patagonia, Argentina, where fascioliasis is endemic. Measures of F. hepatica found in L. europaeus from northern Patagonia demonstrate that the liver fluke is able to fully develop in wild hares and to shed normal eggs through their faeces. Egg shedding to the environment is close to the lower limit obtained for pigs, a domestic animal whose epidemiological importance in endemic areas has already been highlighted. The former, combined with the high prevalence found (14.28%), suggest an even more important role in the transmission cycle than previously considered. The results obtained do not only remark the extraordinary plasticity and adaptability of this trematode species to different host species, but also highlight the role of the European brown hare, and other NIS, as reservoirs capable for parasite spillback to domestic and native cycle, representing a potentially important, but hitherto neglected, cause of disease emergence.

  14. TGF-β superfamily members from the helminth Fasciola hepatica show intrinsic effects on viability and development.

    PubMed

    Japa, Ornampai; Hodgkinson, Jane E; Emes, Richard D; Flynn, Robin J

    2015-03-11

    The helminth Fasciola hepatica causes fasciolosis throughout the world, a major disease of livestock and an emerging zoonotic disease in humans. Sustainable control mechanisms such as vaccination are urgently required. To discover potential vaccine targets we undertook a genome screen to identify members of the transforming growth factor (TGF) family of proteins. Herein we describe the discovery of three ligands belonging to this superfamily and the cloning and characterisation of an activin/TGF like molecule we term FhTLM. FhTLM has a limited expression pattern both temporally across the parasite stages but also spatially within the worm. Furthermore, a recombinant form of this protein is able to enhance the rate (or magnitude) of multiple developmental processes of the parasite indicating a conserved role for this protein superfamily in the developmental biology of a major trematode parasite. Our study demonstrates for the first time the existence of this protein superfamily within F. hepatica and assigns a function to one of the three identified ligands. Moreover further exploration of this superfamily may yield future targets for diagnostic or vaccination purposes due to its stage restricted expression and functional role.

  15. Fasciola hepatica infection reduces Mycobacterium bovis burden and mycobacterial uptake and suppresses the pro-inflammatory response.

    PubMed

    Garza-Cuartero, L; O'Sullivan, J; Blanco, A; McNair, J; Welsh, M; Flynn, R J; Williams, D; Diggle, P; Cassidy, J; Mulcahy, G

    2016-07-01

    Bovine tuberculosis (BTB), caused by Mycobacterium bovis, has an annual incidence in cattle of 0.5% in the Republic of Ireland and 4.7% in the UK, despite long-standing eradication programmes being in place. Failure to achieve complete eradication is multifactorial, but the limitations of diagnostic tests are significant complicating factors. Previously, we have demonstrated that Fasciola hepatica infection, highly prevalent in these areas, induced reduced sensitivity of the standard diagnostic tests for BTB in animals co-infected with F. hepatica and M. bovis. This was accompanied by a reduced M. bovis-specific Th1 immune response. We hypothesized that these changes in co-infected animals would be accompanied by enhanced growth of M. bovis. However, we show here that mycobacterial burden in cattle is reduced in animals co-infected with F. hepatica. Furthermore, we demonstrate a lower mycobacterial recovery and uptake in blood monocyte-derived macrophages (MDM) from F. hepatica-infected cattle which is associated with suppression of pro-inflammatory cytokines and a switch to alternative activation of macrophages. However, the cell surface expression of TLR2 and CD14 in MDM from F. hepatica-infected cattle is increased. These findings reflecting the bystander effect of helminth-induced downregulation of pro-inflammatory responses provide insights to understand host-pathogen interactions in co-infection.

  16. Fasciola hepatica and Paramphistomum daubneyi: field observations on the transport and outcome of floating metacercariae in running water.

    PubMed

    Rondelaud, D; Vignoles, P; Vareille-Morel, C; Abrous, M; Mage, C; Mouzet, R; Dreyfuss, G

    2004-06-01

    Experimental investigations in eight open drainage ditches and furrows from central France were carried out to analyse the dispersal of floating metacercariae of two digenean species by running water and to determine the outcome of larvae which settled on Nasturtium officinale (watercress). The frequencies of larvae found after their transport by water ranged from 33% to 49.7%, thus indicating that more than half of the metacercariae used in this experiment had fallen to the bottom of the water during this transport. The nature of the site (furrow, or ditch supplied by a spring) had a significant effect on the distribution of floating larvae, while the digenean species had no effect. Low percentages of metacercariae on watercress were noted in furrows (3.5-4.3% of larvae) and ditches (0.8-1.3%). When the watercress grew, most larvae that had settled on leaves and stems died but there were always several living metacercariae on this plant (0.7-1.5% of larvae for Fasciola hepatica and 0.2-0.5% for Paramphistomum daubneyi). The weak buoyancy of these floating cysts on running water limited their dispersal and, consequently, led to a real diminution of risks incurred by definitive hosts towards these metacercariae.

  17. Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection of Fasciola gigantica circulating fatty acid binding protein.

    PubMed

    Anuracpreeda, Panat; Chawengkirttikul, Runglawan; Sobhon, Prasert

    2016-09-01

    Up to now, parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Hence, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. In the present study, a monoclonal antibody (MoAb) against recombinant Fasciola gigantica fatty acid binding protein (rFgFABP) has been produced. As well, a reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) has been developed for the detection of circulating FABP in the sera of mice experimentally and cattle naturally infected with F. gigantica. MoAb 3A3 and biotinylated rabbit anti-recombinant FABP antibody were selected due to their high reactivities and specificities. The lower detection limit of sandwich ELISA was 5 pg mL-1, and no cross-reaction with other parasite antigens was observed. This assay could detect F. gigantica infection from day 1 post infection. In experimental mice, the sensitivity, specificity and accuracy of this assay were 93·3, 100 and 98·2%, while in natural cattle they were 96·7, 100 and 99·1%. Hence, this sandwich ELISA method showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica. PMID:27312522

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

    PubMed

    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.

  19. Mechanisms of liver fibrosis associated with experimental Fasciola hepatica infection: roles of Fas2 proteinase and hepatic stellate cell activation.

    PubMed

    Marcos, Luis A; Terashima, Angélica; Yi, Pedro; Andrade, Roy; Cubero, Francisco J; Albanis, Efsevia; Gotuzzo, Eduardo; Espinoza, Jose R; Friedman, Scott L

    2011-02-01

    We have evaluated the possible mechanisms of liver fibrosis caused by Fasciola hepatica in an animal model and in culture using immortalized human stellate cells. Liver biopsies of F. hepatica-infected rats were performed at wk 8 and 16. Serum-starved LX-2 cells, a human stellate cell line, were exposed to increasing concentrations of Fas2 antigen. The expression of key fibrosis-related genes was evaluated by qRT-PCR. There was a significant correlation between fibrogenic gene expression and both intensity and duration of infection. LX-2 cells exposed to Fas2 showed progressively increased expression of mRNAs for Collagen I, alpha-smooth muscle-actin, platelet-derived growth factor beta receptor, and tissue inhibitor of metalloproteinase II; inhibition of Fas2 cysteine proteinase activity by E-64 abrogated these increases, suggesting that the protease activity of Fas2 is involved in fibrogenic stimulation. In summary, F. hepatica infection is associated with up-regulation of mRNAs associated with hepatic fibrogenesis in vivo and in activated hepatic stellate cells.

  20. Fasciola hepatica infection reduces Mycobacterium bovis burden and mycobacterial uptake and suppresses the pro-inflammatory response.

    PubMed

    Garza-Cuartero, L; O'Sullivan, J; Blanco, A; McNair, J; Welsh, M; Flynn, R J; Williams, D; Diggle, P; Cassidy, J; Mulcahy, G

    2016-07-01

    Bovine tuberculosis (BTB), caused by Mycobacterium bovis, has an annual incidence in cattle of 0.5% in the Republic of Ireland and 4.7% in the UK, despite long-standing eradication programmes being in place. Failure to achieve complete eradication is multifactorial, but the limitations of diagnostic tests are significant complicating factors. Previously, we have demonstrated that Fasciola hepatica infection, highly prevalent in these areas, induced reduced sensitivity of the standard diagnostic tests for BTB in animals co-infected with F. hepatica and M. bovis. This was accompanied by a reduced M. bovis-specific Th1 immune response. We hypothesized that these changes in co-infected animals would be accompanied by enhanced growth of M. bovis. However, we show here that mycobacterial burden in cattle is reduced in animals co-infected with F. hepatica. Furthermore, we demonstrate a lower mycobacterial recovery and uptake in blood monocyte-derived macrophages (MDM) from F. hepatica-infected cattle which is associated with suppression of pro-inflammatory cytokines and a switch to alternative activation of macrophages. However, the cell surface expression of TLR2 and CD14 in MDM from F. hepatica-infected cattle is increased. These findings reflecting the bystander effect of helminth-induced downregulation of pro-inflammatory responses provide insights to understand host-pathogen interactions in co-infection. PMID:27108767

  1. Immuno-diagnosis of bubaline fasciolosis with Fasciola gigantica cathepsin-L and recombinant cathepsin L 1-D proteases.

    PubMed

    Raina, O K; Yadav, S C; Sriveny, D; Gupta, S C

    2006-05-01

    Fasciola gigantica cathepsin-L cysteine proteinase and recombinant cathepsin L 1-D were assessed for their potential in the immuno-diagnosis of F. gigantica infection in buffaloes. A diagnostic ELISA, based on these two antigens, was developed to detect antibodies against F. gigantica in water buffaloes. Sensitivity of the ELISA was assessed using sera from buffaloes experimentally or naturally infected with F. gigantica from F. gigantica endemic areas and its specificity by probing the sera of the host from F. gigantica non-endemic area. Our earlier studies under experimental setting showed 100% sensitivity of cathepsin-L ELISA in the diagnosis of fasciolosis in buffaloes, with the earliest detection of infection at 4 weeks post-infection. However, under field situation of natural F. gigantica infection, this sensitivity declined to 97.1% but specificity of the test remained 100%. Cross-reactivity of the antigen was checked with Schistosoma indicum, S. spindale, Paramphistomum epiclitum, Gastrothylax spp., Gigantocotyle explanatum, hydatid and Strongyloides papilossus in the bubaline host, naturally infected with these helminths. F. gigantica cathepsin-L and the recombinant cathepsin L-1D does not cross-react with these helminth parasites in natural mono or mixed infection of the host. The present ELISA contributes a relatively sensitive and reliable tool for the early serodiagnosis of bubaline fasciolosis.

  2. Biological control of Fasciola hepatica eggs with the Pochonia chlamydosporia fungus after passing through the cattle gastrointestinal tract.

    PubMed

    Dias, Anderson S; Araújo, Jackson V; Braga, Fábio R; Araujo, Juliana M; Puppin, André C; Fernandes, Fernanda M; Ramos, Rafael F; Bertonceli, Raul M; da Silva, Renata G; Perboni, Wilber R

    2012-02-01

    Fasciolosis is a disease caused by Fasciola hepatica responsible for causing significant losses in livestock. This study aimed to evaluate the Pochonia chlamydosporia fungus (isolate VC1) on F. hepatica eggs after passing through the cattle gastrointestinal tract. For this evaluation, 1 g pellet was given in sodium alginate matrix per kilogram live weight containing 25% of fungal mycelium from isolate VC1 per animal. Twelve animals were used, six treated and six untreated (control). Some stool samples were collected from the groups of treated and control animals, at the times of 12, 18, 24, 48, 72, and 96 h after the pellets' administration. Then, from each stool sample of treated and control groups, 2 g was placed in a Petri dish of 9 cm in diameter, containing 2% water-agar and 1,000 eggs of F. hepatica. It was observed that the fungus was effective in preying upon the eggs in the samples recovered at all of the schedules starting at 12 h. Furthermore, differences were observed (p < 0.01) in the destruction of eggs in the Petri dishes in the treated group compared with the control group. The ovicidal effect was observed after 7 days of interaction. The ovicidal P. chlamydosporia fungus was effective in destroying F. hepatica eggs; therefore, it is suggested that this fungus could be employed as agent for the control of helminth eggs. PMID:21773773

  3. An analysis of the calcium-binding protein 1 of Fasciola gigantica with a comparison to its homologs in the phylum Platyhelminthes.

    PubMed

    Vichasri-Grams, Suksiri; Subpipattana, Pornpimol; Sobhon, Prasert; Viyanant, Vithoon; Grams, Rudi

    2006-03-01

    A full-length cDNA encoding the Fasciola gigantica calcium-binding protein 1 (FgCaBP1) was cloned from an adult stage cDNA expression library in an immunoscreen using rabbit immune serum against the parasite's excretion/secretion antigens. The deduced amino acid sequence showed 96.3% identity to Fh22CBP of Fasciola hepatica. During development in the mammalian host FgCaBP1 RNA was detected in metacercariae, juveniles and adults and was exclusively localized to the tegumental cell bodies. Immune serum of a rabbit infected with F. gigantica detected recombinant FgCaBP1 starting from the sixth week of infection. Immune sera of mice infected with Schistosoma mansoni and Schistosoma mekongi cross-reacted with recombinant FgCaBP1 in immunoblots. Recombinant FgCaBP1 showed calcium and magnesium-binding activity by a mobility shift during non-denaturing PAGE in the presence of Ca2+ or Mg2+, respectively. A polyclonal mouse anti-rFgCaBP1 antiserum detected the native protein as a major component of the parasite's tegumental antigens in immunoblots and as a strictly tegumental antigen in tissue cross-sections of adult and juvenile parasites. Comparative sequence analysis of homologs from Fasciola and Schistosoma present in the GenBank database revealed sequence signatures specific to these trematode proteins and thereby indicates their origin from a single ancestor. FgCaBP1 contains two adjacent, N-terminal located EF-hands and a C-terminal located domain similar to dynein light chain type 1. Independent structure predictions of the two domains suggest that they will fold according to the already determined structures of the EF-hand motif and the dynein light chain type 1 proteins.

  4. Comparing an in vivo egg reduction test and in vitro egg hatching assay for different anthelmintics against Fasciola species, in cattle.

    PubMed

    Arafa, Waleed M; Shokeir, Khalid M; Khateib, Abdelrahman M

    2015-11-30

    This study aimed to compare between the efficiency of in vivo fecal egg reduction test (FERT) and in vitro egg hatching assay (EHA) in evaluating of the anti-Fasciola activity of albendazole, triclabendazole, oxyclozanide and praziquantel. A field trial was carried out on fifty naturally Fasciola infected cattle that were divided equally into 5 groups (A-E). On day zero; groups A-D were drenched with albendazole, triclabendazole, oxyclozanide or praziquantel, respectively, while the remaining one, group E, was kept as untreated control. Fecal egg counts of the different groups were conducted weekly over a period of one month post-treatment. In vitro, commercial albendazole and oxyclozanide were diluted to 0.0002, 0.002, 0.02, 0.2 and 2.0 μg/ml, while commercial triclabendazole and praziquantel were diluted to concentrations of 25, 50, 75 and 100 μg/ml with dimethyl sulfoxide (DMSO). In vivo, at the 2nd week post-treatment, triclabendazole and oxyclozanide showed 100% fecal egg reduction (FER), and albendazole had a maximum of 73.7% reduction (P < 0.0001), however, praziquantel did not record any reduction of Fasciola egg counts. In vitro, triclabendazole treated Fasciola gigantica eggs showed early embryonic lysis with zero% hatching at the different concentrations (P < 0.01). In albendazole, the hatching varied according to the drug concentration. At the highest two concentrations; 0.2 and 2.0 μg/ml, the hatching percentages were 7.4 ± 1.6 and 5.6 ± 1.5 (P < 0.01) respectively. On the contrary, there were no significant differences in egg development and hatching percentage of oxyclozanide or praziquantel treated groups. In conclusion, the efficacy of triclabendazole and albendazole as fasciolicdes could be predicted by Egg Hatching Assay (EHA). Meanwhile fasciolicide activity of oxyclozanide could not be assessed with EHA. Based on in vivo and in vitro findings, paraziquantel did not show any fasciolicide effect.

  5. Bakkeius moragai n. gen. et n. sp. and Pojmanskia riosae n. gen. et n. sp. (Trematoda: Digenea: Brachylaimoidea) in birds from the Area de Conservación Guanacaste, Costa Rica.

    PubMed

    Zamparo, David; Brooks, Daniel R; Causey, Douglas

    2003-08-01

    We describe 2 new species of leucochloridiid-like brachylaimoid digeneans parasitizing a variety of birds in the Area de Conservación Guanacaste, Costa Rica, each of which we assign to a new genus. According to Pojmanska's (Pojmanska, T. 2002a. Superfamily Brachylaimoidea Joyeux & Foley, 1930. In Keys to the Trematoda, D. I. Gibson, A. Jones, and R. A. Bray [eds.]. CAB International and The Natural History Museum, London, U.K., p. 31-36.) key for the Brachylaimoidea, we are unable to place either species in any family. One species most closely resembles members of Leucochloridium by having well-developed suckers, lacking an esophagus, and having cecal shoulders, gonads at the posterior end, and the genital pore at posterior end of body but differs by having symmetrical testes, a posttesticular ovary, and a terminal genital pore; thus, we propose the genus Bakkeius for it. The second new genus resembles members of Michajlovia by having ventral genital pores but differs by having extracecal uterine loops in the forebody, a cirrus sac containing the pars prostatica and seminal vesicle, and gland cells surrounding the genital pore; thus, we propose Pojmanskia for it. These new genera must currently be treated as incertae sedis according to Pojmanska (op. cit.); however, we feel that future phylogenetic analyses will require emendation of the family diagnosis for Leucochloridiidae to include those taxa with terminal and ventral genital pores and with preovarian testes.

  6. Immunodiagnosis of Fasciola hepatica infection (fascioliasis) in a human population in the Bolivian Altiplano using purified cathepsin L cysteine proteinase.

    PubMed

    O'Neill, S M; Parkinson, M; Strauss, W; Angles, R; Dalton, J P

    1998-04-01

    Cathepsin L1 (CL1), an immunogenic cysteine proteinase secreted by juvenile and adult Fasciola hepatica, was assessed for its potential as a diagnostic agent for the serologic detection of human fascioliasis. Using ELISAs, we compared the ability of liver fluke homogenates (LFH), excretory/secretory (ES) products, and CL1 to discriminate between seropositive (infected) and seronegative (noninfected) individuals within a population of 95 patients from the Bolivian Altiplano. A high prevalence of human fascioliasis has been reported in this region. The division between the seropositive and seronegative individuals was poorly defined when LFH was used as the antigen. A greater discrimination between these populations was achieved with both ES and CL1. A K-means cluster analysis using the combined ES and CL1 ELISA data identified a cluster of seropositive individuals. Cathepsin L1 detected a subset (20) of these seropositive individuals while ES detected all 26; however, ES detected nine additional individuals that were in the seronegative cluster. The ratio of the mean absorbance readings between seropositive and seronegative individuals was markedly improved by using conjugated second antibodies to IgG4, the predominant isotype elicited by infection. In these IgG4-ELISAs, CL1 again identified fewer individuals as seropositive than did ES, but improved the discrimination between the seropositive and seronegative individuals and thus provided a more conclusive diagnosis. Sera obtained from patients infected with schistosomiasis mansoni, cysticercosis, hydatidosis, and Chagas' disease were negative in these assays, which demonstrated the specificity of the IgG4-ELISA for detecting fascioliasis. Twenty of the 95 patients (21%) were seropositive for fascioliasis by the CL1 IgG4-ELISA, confirming the earlier reports of the high prevalence of disease in this region. A standardized diagnostic test for human fascioliasis, based on an ELISA that detects IgG4 responses to CL1

  7. Relationships between host species and morphometric patterns in Fasciola hepatica adults and eggs from the northern Bolivian Altiplano hyperendemic region.

    PubMed

    Valero, M A; Darce, N A; Panova, M; Mas-Coma, S

    2001-12-01

    The highest prevalences and intensities of human fasciolosis by Fasciola hepatica are found in the northern Bolivian Altiplano, where sheep and cattle are the main reservoir host species and pigs and donkeys the secondary ones. Morphometric comparisons of many linear measurements, areas and ratios of F. hepatica adults (from sheep, cattle and pigs) and eggs (from sheep, cattle, pigs and donkeys) in natural liver fluke populations of the Bolivian Altiplano, as well as of F. hepatica adults and eggs experimentally obtained in Wistar rats infected with Altiplanic sheep, cattle and pig isolates, were made using computer image analysis and an allometric model. Although morphometric values of adult flukes from natural populations of sheep, cattle, and pigs showed great overlap, there were clear differences in allometric growth. The allometries analyzed were: body area (BA) versus body length (BL), BA versus body width (BW), BA versus perimeter (Pe), BA versus distance between posterior end of body and ventral sucker (P-VS), BL versus BW, BL versus Pe, and BL versus P-VS. These allometries show a good fit in the seven pairs of variables in all the populations examined. Comparative statistical analysis of the allometries shows that fluke adult populations from sheep, cattle and pigs significantly differ in BL versus BW and BL versus P-VS functions. Statistical analysis of F. hepatica egg size shows characteristic morphometric traits in each definitive host species. In experimentally infected rats, fluke adult allometry and egg morphometry do not vary depending on the Altiplanic definitive host species isolate. Our study reveals that the definitive host species decisively influences the size of F. hepatica adults and eggs, and these influences do not persist in a rodent definitive host model.

  8. Fasciola hepatica: tegumental alterations in adult flukes following in vitro and in vivo administration of artesunate and artemether.

    PubMed

    Keiser, Jennifer; Morson, Gianni

    2008-02-01

    The tegumental changes in adult Fasciola hepatica induced by artemether and artesunate were assessed utilizing scanning electron microscopy (SEM). F. hepatica were incubated with artemether and artesunate for 48h at a concentration of 10microg/ml in the absence or presence of haemin. For the latter experiment both, a triclabendazole-resistant and sensitive F. hepatica isolate were used. For the in vivo studies rats were treated with single 200mg/kg oral doses of artemether and artesunate and flukes recovered from the bile ducts after 24-96h. SEM analysis of the flukes incubated in the presence of the drugs without haemin showed only minor and localized damage of the tegument. In the presence of haemin extensive tegumental damage, including sloughing, blebbing and eruptions, particularly in the ventral and dorsal mid-body and tail region, was evident. No difference in the extent of damage could be observed between artemether and artesunate and between the triclabendazole-resistant and non-resistant flukes. After 24h in vivo disruption of the tegument was evident in the artemether-treated flukes, and the damage increased in severity 48-72h post-treatment. Sloughing, swelling and extensive furrowing of the tegument was observed in several flukes, in particular in the tail region and the ventral apical cone region. In the artesunate treatment, tegumental damage was evident after 72h, but seemed slightly less pronounced when compared to the artemether-treated specimens examined at the same time point. Concluding our experiments confirm that artemether and artesunate are potent fasciocidal drugs and the tegument of adult F. hepatica appears to be a target for the action of these drugs.

  9. Fasciola gigantica: enzymes of the ornithine-proline-glutamate pathway--characterization of delta1-pyrroline-5-carboxylate dehydrogenase.

    PubMed

    Mohamed, Saleh A; Mohamed, Tarek M; Fahmy, Afaf S; El-Badry, Mohamed O; Abdel-Gany, Somia S

    2008-01-01

    Ornithine aminotransferase (OAT), proline oxidase (PO), Delta 1-pyrroline-5-carboxylate reductase (P5CR), and Delta 1-pyrroline-5-carboxylate dehydrogenase (P5CD) were assessed in Fasciola gigantica. All enzymes are involved in the conversion of ornithine into glutamate and proline. High levels of P5CD suggest that the direction of the metabolic flow from ornithine is more toward glutamate than proline. F. gigantica P5CD1 and P5CD2 were separated from the majority of contaminating proteins in crude homogenate using a CM-cellulose column. A Sephacryl S-200 column was employed for P5CD2 to obtain pure enzyme with increased specific activity. The molecular mass of P5CD2 was estimated to be 50kDa using a Sephacryl S-200 column and SDS-PAGE. It migrated as a single band on SDS-PAGE, indicating a monomeric enzyme. P5CD2 had Km values of 1.44mM and 0.37mM for NAD and P5C, respectively. P5CD2 oxidized a number of aliphatic and aromatic aldehydes, where the aromatic compounds had higher affinity toward the enzyme. All amino acids examined had partial inhibitory effects on the enzyme. While 3mM AMP caused 31% activation of enzyme, 3mM ADP and ATP inhibited activity by 18% and 23%, respectively. Apart from Cu2+, the divalent cations that were studied caused partial inhibitory effects on the enzyme. PMID:17655846

  10. Approach to molecular characterization of different strains of Fasciola hepatica using random amplified polymorphic DNA polymerase chain reaction.

    PubMed

    Scarcella, S; Miranda-Miranda, E; Solana, M V; Solana, H

    2015-04-01

    The aim of the present study was to genetically characterize Fasciola hepatica strains from diverse ecogeographical regions (America and Europe), susceptible and resistant to Triclabendazole, using the random amplified polymorphic DNA fragments (RAPDs-PCR) technique to elucidate genetic variability between the different isolates. Ten different oligonucleotide primers of 10 bases with GC content varying from 50-70% were used. A polymerase chain reaction (PCR) was carried out in 25 μl of total volume. Duplicate PCR reactions on each individual template DNA were performed to test the reproducibility of the individual DNA bands. The size of the RAPD-PCR fragments was determined by the reciprocal plot between the delay factors (Rf) versus the logarithm of molecular weight ladder. The phenogram obtained showed three main clusters, the major of which contained European Strains (Cullompton and Sligo) showing a genetic distance of 27.2 between them. The American strains (Cedive and Cajamarca) on the other hand formed each their distinctive group but clearly maintaining a closer genetic relationship among them than that to their European counterparts, with which showed a distance of 33.8 and 37.8, respectively. This polymorphism would give this species enhanced adaptability against the host, as well as the environment. The existence of genetically different populations of F. hepatica could allow, against any selection pressure, natural or artificial (for use fasciolicides products and/or control measures), one or more populations of F. hepatica to be able to survive and create resistance or adaptability to such selective pressure. PMID:25595655

  11. Relation between iron metabolism and antioxidants enzymes and δ-ALA-D activity in rats experimentally infected by Fasciola hepatica.

    PubMed

    Bottari, Nathieli B; Mendes, Ricardo E; Baldissera, Matheus D; Bochi, Guilherme V; Moresco, Rafael N; Leal, Marta L R; Morsch, Vera M; Schetinger, Maria R C; Christ, Ricardo; Gheller, Larissa; Marques, Éder J; Da Silva, Aleksandro S

    2016-06-01

    The aim of this study was to evaluate the iron metabolism in serum, as well as antioxidant enzymes, in addition to the Delta-Aminolevulinic Acid Dehydratase (δ-ALA-D) activity in the liver of rats experimentally infected by Fasciola hepatica. Thirty male adult rats (Wistar) specific pathogen free were divided into four groups: two uninfected group (CTRL 1 and CTRL 2) with five animals each and two infected groups (INF 1 and INF 2) with 10 animals each. Infection was performed orally with 20 metacercariae at day 1. On day 15 (CTRL 1 and INF 1 groups) and 87 PI (CTRL 2 and INF 2 groups) blood and bone marrow were collected and the animals were subsequently euthanized for liver sampling. Blood was allocated in tubes without anticoagulant for serum acquisition to measure iron, transferrin and unsaturated iron binding capacity (UIBC). δ-ALA-D, superoxide dismutase (SOD), and catalase (CAT) activities were measured in the liver. A decrease in iron, transferrin and UIBC levels was observed in all infected animals compared to control groups (P < 0.05). Furthermore, iron accumulation was observed in bone marrow of infected mice. Infected animals showed an increase in δ-ALA-D activity at 87 post-infection (PI) (INF 2) as well as in SOD activity at days 15 (INF 1) and 87 PI (INF 2). On the other hand, CAT activity was reduced in rats infected by F. hepatica during acute and chronic phase of fasciolosis (INF 1 and INF 2 groups), when moderate (acute) and severe necrosis in the liver histopathology were observed. These results may suggest that oxidative damage to tissues along with antioxidant mechanisms might have taken part in fasciolosis pathogenesis and are also involved in iron deficiency associated to changes in δ-ALA-D activity during chronic phase of disease.

  12. Elucidating the transcriptome of Fasciola hepatica - a key to fundamental and biotechnological discoveries for a neglected parasite.

    PubMed

    Young, Neil D; Hall, Ross S; Jex, Aaron R; Cantacessi, Cinzia; Gasser, Robin B

    2010-01-01

    Liver flukes of animals are parasitic flatworms (Platyhelminthes: Digenea) of major socioeconomic importance in many countries. Key representatives, such as Fasciola hepatica and F. gigantica, cause "liver fluke disease" (= fascioliasis), which is of major animal health significance worldwide. In particular, F. hepatica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. This parasite is also a major food-borne pathogen of humans throughout parts of the Middle East, Asia and South America. Currently, there is a significant focus on the development of new approaches for the prevention and control of fascioliasis in livestock. Recent technological advances in genomics and bioinformatics provide unique opportunities for the identification and prevalidation of drug targets and vaccines through a better understanding of the biology of F. hepatica and related species as well as their relationship with their hosts at the molecular level. Surprisingly, despite the widespread socioeconomic impact of fascioliasis, genomic datasets for F. hepatica are scant, limiting the molecular biological research of this parasite. The present article explores specifically the transcriptome of the adult stage of F. hepatica using an integrated genomic-bioinformatic platform. The analysis of the current data reveals numerous molecules of biological relevance, some of which are inferred to be involved in key biological processes or pathways that could serve as targets for new trematocidal drugs or vaccines. Improved insights into the transcriptome of F. hepatica should pave the way for future, comparative analysis of the transcriptomes of other developmental stages of this and related parasites, such as F. gigantica, cancer-causing flatworms (Clonorchis sinensis and Opisthorchis viverrini) and blood flukes (Schistosoma mansoni and S. japonicum). Prediction of the

  13. MOLLUSCICIDAL AND LARVICIDAL ACTIVITIES OF Atriplex inflata AERIAL PARTS AGAINST THE MOLLUSK Galba truncatula, INTERMEDIATE HOST OF Fasciola hepatica.

    PubMed

    Hamed, Néjia; Njeh, Fatma; Damak, Mohamed; Ayadi, Ali; Mezghani-Jarraya, Raoudha; Hammami, Hayet

    2015-12-01

    Fasciolosis is a widespread parasitosis of farm live-stock in many developing countries. For this reason, it is necessary to search for new substances against parasitic diseases caused by flukes. Indeed, a wide variety of terrestrial plants have been subjected to chemical and pharmacological screening in order to discover their potential for human medicinal use. The molluscicidal and larvicidal activities of Atriplex inflata were tested on Galba truncatula and Fasciola hepatica larval stages infecting this snail in Tunisia. Phytochemical tests were conducted on extracts in order to establish a meaningful relationship with molluscicidal and larvicidal activities. The molluscicidal activity was evaluated by subjecting snails to sample aqueous solutions. Accordingly, hexane, ethyl acetate, methanol and methanol-water (8:2, v-v) were used as extraction solvents. As a result, hexane and ethyl acetate extracts showed potent activity, according to the World Health Organization, giving LC50 = 7.59 mg/L and 6.69 mg/L for hexane extracts of leaves and fruits, respectively. Ethyl acetate extracts gave LC50 = 5.90 mg/L and 7.32 mg/L for leaves and fruits, successively. Molluscicidal activities of powders were less potent on snails, but active according to the World Health Organization. Hexane and ethyl acetate extracts from leaves and fruits gave potent larvicidal activities with a delay rate exceeding 45.50% (45.50- 98.92%). Phytochemical tests showed that these activities may be attributed to the presence of triterpenoids and/or sterols. PMID:27049700

  14. Glycans from Fasciola hepatica Modulate the Host Immune Response and TLR-Induced Maturation of Dendritic Cells.

    PubMed

    Rodríguez, Ernesto; Noya, Verónica; Cervi, Laura; Chiribao, María Laura; Brossard, Natalie; Chiale, Carolina; Carmona, Carlos; Giacomini, Cecilia; Freire, Teresa

    2015-12-01

    Helminths express various carbohydrate-containing glycoconjugates on their surface, and they release glycan-rich excretion/secretion products that can be very important in their life cycles, infection and pathology. Recent evidence suggests that parasite glycoconjugates could play a role in the evasion of the immune response, leading to a modified Th2-polarized immune response that favors parasite survival in the host. Nevertheless, there is limited information about the nature or function of glycans produced by the trematode Fasciola hepatica, the causative agent of fasciolosis. In this paper, we investigate whether glycosylated molecules from F. hepatica participate in the modulation of host immunity. We also focus on dendritic cells, since they are an important target of immune-modulation by helminths, affecting their activity or function. Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production. During infection, this parasite is able to induce a semi-mature phenotype of DCs expressing low levels of MHCII and secrete IL-10. Furthermore, we show that parasite glycoconjugates mediate the modulation of LPS-induced maturation of DCs since their oxidation restores the capacity of LPS-treated DCs to secrete high levels of the pro-inflammatory cytokines IL-6 and IL-12/23p40 and low levels of the anti-inflammatory cytokine IL-10. Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2. The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis. PMID:26720149

  15. Approach to molecular characterization of different strains of Fasciola hepatica using random amplified polymorphic DNA polymerase chain reaction.

    PubMed

    Scarcella, S; Miranda-Miranda, E; Solana, M V; Solana, H

    2015-04-01

    The aim of the present study was to genetically characterize Fasciola hepatica strains from diverse ecogeographical regions (America and Europe), susceptible and resistant to Triclabendazole, using the random amplified polymorphic DNA fragments (RAPDs-PCR) technique to elucidate genetic variability between the different isolates. Ten different oligonucleotide primers of 10 bases with GC content varying from 50-70% were used. A polymerase chain reaction (PCR) was carried out in 25 μl of total volume. Duplicate PCR reactions on each individual template DNA were performed to test the reproducibility of the individual DNA bands. The size of the RAPD-PCR fragments was determined by the reciprocal plot between the delay factors (Rf) versus the logarithm of molecular weight ladder. The phenogram obtained showed three main clusters, the major of which contained European Strains (Cullompton and Sligo) showing a genetic distance of 27.2 between them. The American strains (Cedive and Cajamarca) on the other hand formed each their distinctive group but clearly maintaining a closer genetic relationship among them than that to their European counterparts, with which showed a distance of 33.8 and 37.8, respectively. This polymorphism would give this species enhanced adaptability against the host, as well as the environment. The existence of genetically different populations of F. hepatica could allow, against any selection pressure, natural or artificial (for use fasciolicides products and/or control measures), one or more populations of F. hepatica to be able to survive and create resistance or adaptability to such selective pressure.

  16. A 170kDa multi-domain cystatin of Fasciola gigantica is active in the male reproductive system.

    PubMed

    Geadkaew, Amornrat; Kosa, Nanthawat; Siricoon, Sinee; Grams, Suksiri Vichasri; Grams, Rudi

    2014-09-01

    Cystatins are functional as intra- and extracellular inhibitors of cysteine proteases and are expressed as single or multi-domain proteins. We have previously described two single domain type 1 cystatins in the trematode Fasciola gigantica that are released into the parasite's intestinal tract and exhibit inhibitory activity against endogenous and host cathepsin L and B proteases. In contrast, the here presented 170kDa multi-domain cystatin (FgMDC) comprises signal peptide and 12 tandem repeated cystatin-like domains with similarity to type 2 single domain cystatins. The domains show high sequence divergence with identity values often <20% and at only 26.8% between the highest matching domains 6 and 10. Several domains contain degenerated QVVAG core motifs and/or lack other important residues of active type 2 cystatins. Domain-specific antisera detected multiple forms of FgMDC ranging from <10 to >120kDa molecular mass in immunoblots of parasite crude extracts and ES product with different banding patterns for each antiserum demonstrating complex processing of the proprotein. The four domains with the highest conserved QVVAG motifs were expressed in Escherichia coli and the refolded recombinant proteins blocked cysteine protease activity in the parasite's ES product. Strikingly, immunohistochemical analysis using seven domain-specific antisera localized FgMDC in testis lobes and sperm. It is speculated that the processed cystatin-like domains have function analogous to the mammalian group of male reproductive tissue-specific type 2 cystatins and are functional in spermiogenesis and fertilization.

  17. Fasciola hepatica mucin-encoding gene: expression, variability and its potential relevance in host-parasite relationship.

    PubMed

    Cancela, Martín; Santos, Guilherme B; Carmona, Carlos; Ferreira, Henrique B; Tort, José Francisco; Zaha, Arnaldo

    2015-12-01

    Fasciola hepatica is the causative agent of fasciolosis, a zoonosis with significant impact both in human and animal health. Understanding the basic processes of parasite biology, especially those related to interactions with its host, will contribute to control F. hepatica infections and hence liver pathology. Mucins have been described as important mediators for parasite establishment within its host, due to their key roles in immune evasion. In F. hepatica, mucin expression is upregulated in the mammalian invasive newly excysted juvenile (NEJ) stage in comparison with the adult stage. Here, we performed sequencing of mucin cDNAs prepared from NEJ RNA, resulting in six different cDNAs clusters. The differences are due to the presence of a tandem repeated sequence of 66 bp encoded by different exons. Two groups of apomucins one with three and the other with four repeats, with 459 and 393 bp respectively, were identified. These cDNAs have open reading frames encoding Ser-Thr enriched proteins with an N-terminal signal peptide, characteristic of apomucin backbone. We cloned a 4470 bp gene comprising eight exons and seven introns that encodes all the cDNA variants identified in NEJs. By real time polymerase chain reaction and high-resolution melting approaches of individual flukes we infer that fhemuc-1 is a single-copy gene, with at least two different alleles. Our data suggest that both gene polymorphism and alternative splicing might account for apomucin variability in the fhemuc-1 gene that is upregulated in NEJ invasive stage. The relevance of this variation in host-parasite interplay is discussed.

  18. Ultrastructural changes in the tegument and gut of adult Fasciola hepatica following in vivo treatment with artesunate.

    PubMed

    O'Neill, J F; Johnston, R C; Halferty, L; Brennan, G P; Fairweather, I

    2015-07-01

    An in vivo study in the laboratory rat model has been carried out to monitor changes to the tegument and gut of adult Fasciola hepatica following treatment with artesunate. Rats infected with the triclabendazole-resistant Oberon isolate were dosed orally with artesunate at a concentration of 200 mg/kg and flukes recovered 24, 48, 72 and 96 h post-treatment (pt). The flukes were processed for scanning and transmission electron microscope examination. Changes to the external surface were limited to swelling and blebbing of the interspinal tegument. There was one exception, a specimen recovered 72 h pt, which had completely lost the syncytium over the posterior region of the fluke. Internal changes to the tegumental syncytium and cell bodies were more severe and were apparent from 48 h pt onwards. Increased numbers of secretory bodies were present in the apical region of the syncytium, the basal infolds were swollen and sloughing of the apical plasma membrane was seen at 96 h pt. In the cell bodies, there was swelling and vesiculation of the cisternae of the granular endoplasmic reticulum (ger), swelling of the mitochondria and a decrease in secretory body production. Changes to the gastrodermal cells were evident from 24 h onwards. They comprised swelling and vesiculation of the ger cisternae, swelling and lysis of the mitochondria and accumulation of autophagic vacuoles and lipid droplets. The nuclei of the cells were karyopyknotic by 96 h pt. The gut was consistently more severely affected than the tegument at all time points pt, pointing to an oral route of uptake for artesunate. This study has provided information on the primary subcellular targets for drug action in the fluke.

  19. UDP-galactose 4'-epimerase from the liver fluke, Fasciola hepatica: biochemical characterization of the enzyme and identification of inhibitors.

    PubMed

    Zinsser, Veronika L; Lindert, Steffen; Banford, Samantha; Hoey, Elizabeth M; Trudgett, Alan; Timson, David J

    2015-03-01

    Leloir pathway enzyme uridine diphosphate (UDP)-galactose 4'-epimerase from the common liver fluke Fasciola hepatica (FhGALE) was identified and characterized. The enzyme can be expressed in, and purified from, Escherichia coli. The recombinant enzyme is active: the K(m) (470 μM) is higher than the corresponding human enzyme (HsGALE), whereas the k(cat) (2.3 s(-1)) is substantially lower. FhGALE binds NAD(+) and has shown to be dimeric by analytical gel filtration. Like the human and yeast GALEs, FhGALE is stabilized by the substrate UDP-galactose. Molecular modelling predicted that FhGALE adopts a similar overall fold to HsGALE and that tyrosine 155 is likely to be the catalytically critical residue in the active site. In silico screening of the National Cancer Institute Developmental Therapeutics Program library identified 40 potential inhibitors of FhGALE which were tested in vitro. Of these, 6 showed concentration-dependent inhibition of FhGALE, some with nanomolar IC50 values. Two inhibitors (5-fluoroorotate and N-[(benzyloxy)carbonyl]leucyltryptophan) demonstrated selectivity for FhGALE over HsGALE. These compounds also thermally destabilized FhGALE in a concentration-dependent manner. Interestingly, the selectivity of 5-fluoroorotate was not shown by orotic acid, which differs in structure by 1 fluorine atom. These results demonstrate that, despite the structural and biochemical similarities of FhGALE and HsGALE, it is possible to discover compounds which preferentially inhibit FhGALE.

  20. Evidence of Fasciola hepatica infection in Radix peregra and a mollusc of the family Succineidae in Ireland.

    PubMed

    Relf, V; Good, B; McCarthy, E; de Waal, T

    2009-07-01

    Worldwide molluscs of the genera Lymnaea, Pseudosuccinea, Galba and Stagnicola act as intermediate hosts of the common liver fluke, Fasciola hepatica. Galba truncatula is the only recorded intermediate host of F. hepatica within Ireland and is the principle intermediate host within Europe. To investigate the transmission of liver fluke on the Teagasc hill sheep farm, Co. Mayo, Ireland, snail species other than G. truncatula, were collected from the lowland pasture (snail sp. 'A') in May (n=23) and from the hill area (snail sp. 'B') in March, April and May 2008 (n=78, 53 and 36 respectively). Morphological characteristics identified snail sp. 'A' as a Succinea sp. and snail sp. 'B' as Radix peregra. PCR and subsequent sequencing of the 18S gene and internal transcribed spacer 2 (ITS-2) region of the snail rDNA were used for molecular identification. The 18S ribosomal DNA (rDNA) sequences revealed a 99% similarity with Omalonyx matheroni (Succineidae) for snail sp. 'A', and for snail sp. 'B' a 99% and 98% similarity with Radix auricularia and R. peregra, respectively. The ITS-2 sequences of snail sp. 'B' revealed a 100% similarity with R. peregra. Using PCR F. hepatica DNA was identified in 73.9% (n=17) of snail sp. 'A' collected during May and 10.3% (n=8), 60.4% (n=32) and 61.1% (n=22) of snail sp. 'B' collected during March, April and May, respectively. A priori knowledge of habitat preference and environmental tolerances of G. truncatula has failed to explain the prevalence of infection recorded in the hill sheep flock on Teagasc hill sheep farm. These findings may explain the presence of infection in sheep populations grazing hill habitats considered too acidic to maintain G. truncatula populations.

  1. Glycans from Fasciola hepatica Modulate the Host Immune Response and TLR-Induced Maturation of Dendritic Cells.

    PubMed

    Rodríguez, Ernesto; Noya, Verónica; Cervi, Laura; Chiribao, María Laura; Brossard, Natalie; Chiale, Carolina; Carmona, Carlos; Giacomini, Cecilia; Freire, Teresa

    2015-12-01

    Helminths express various carbohydrate-containing glycoconjugates on their surface, and they release glycan-rich excretion/secretion products that can be very important in their life cycles, infection and pathology. Recent evidence suggests that parasite glycoconjugates could play a role in the evasion of the immune response, leading to a modified Th2-polarized immune response that favors parasite survival in the host. Nevertheless, there is limited information about the nature or function of glycans produced by the trematode Fasciola hepatica, the causative agent of fasciolosis. In this paper, we investigate whether glycosylated molecules from F. hepatica participate in the modulation of host immunity. We also focus on dendritic cells, since they are an important target of immune-modulation by helminths, affecting their activity or function. Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production. During infection, this parasite is able to induce a semi-mature phenotype of DCs expressing low levels of MHCII and secrete IL-10. Furthermore, we show that parasite glycoconjugates mediate the modulation of LPS-induced maturation of DCs since their oxidation restores the capacity of LPS-treated DCs to secrete high levels of the pro-inflammatory cytokines IL-6 and IL-12/23p40 and low levels of the anti-inflammatory cytokine IL-10. Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2. The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis.

  2. MOLLUSCICIDAL AND LARVICIDAL ACTIVITIES OF Atriplex inflata AERIAL PARTS AGAINST THE MOLLUSK Galba truncatula, INTERMEDIATE HOST OF Fasciola hepatica

    PubMed Central

    HAMED, Néjia; NJEH, Fatma; DAMAK, Mohamed; AYADI, Ali; MEZGHANI-JARRAYA, Raoudha; HAMMAMI, Hayet

    2015-01-01

    Fasciolosis is a widespread parasitosis of farm live-stock in many developing countries. For this reason, it is necessary to search for new substances against parasitic diseases caused by flukes. Indeed, a wide variety of terrestrial plants have been subjected to chemical and pharmacological screening in order to discover their potential for human medicinal use. The molluscicidal and larvicidal activities of Atriplex inflata were tested on Galba truncatula and Fasciola hepatica larval stages infecting this snail in Tunisia. Phytochemical tests were conducted on extracts in order to establish a meaningful relationship with molluscicidal and larvicidal activities. The molluscicidal activity was evaluated by subjecting snails to sample aqueous solutions. Accordingly, hexane, ethyl acetate, methanol and methanol-water (8:2, v-v) were used as extraction solvents. As a result, hexane and ethyl acetate extracts showed potent activity, according to the World Health Organization, giving LC50 = 7.59 mg/L and 6.69 mg/L for hexane extracts of leaves and fruits, respectively. Ethyl acetate extracts gave LC50 = 5.90 mg/L and 7.32 mg/L for leaves and fruits, successively. Molluscicidal activities of powders were less potent on snails, but active according to the World Health Organization. Hexane and ethyl acetate extracts from leaves and fruits gave potent larvicidal activities with a delay rate exceeding 45.50% (45.50- 98.92%). Phytochemical tests showed that these activities may be attributed to the presence of triterpenoids and/or sterols. PMID:27049700

  3. Efficacy of treatment of cattle for liver fluke at housing: influence of differences in flukicidal activity against juvenile Fasciola hepatica.

    PubMed

    Forbes, A B; Reddick, D; Stear, M J

    2015-03-28

    Flukicides are commonly administered at housing to cattle that have grazed fluke-infected pastures or that have been purchased from endemic areas. The choice of product is determined by numerous factors, one of which is the stages of Fasciola hepatica that are killed. Flukicides can be categorised into three main groups: (A) those that kill all juvenile stages and adults; (B) those that kill juveniles from six to eight weeks of age and adults and (C) those that kill adults only. This study was conducted on a commercial beef farm in Scotland and was designed to compare the efficacy of flukicides from each of these three classes in terms of their effects on faecal egg output, coproantigen and liveweight gain. The majority of animals in the untreated control group were positive for coproantigen, fluke eggs or both throughout the study duration of 16 weeks. Egg reappearance interval following housing treatment was eight weeks for clorsulon and 13 weeks for nitroxynil, though patent infections in both groups developed in only a small minority of animals; no fluke eggs were recovered from cattle treated with triclabendazole. Coproantigen was detected four weeks before the reappearance of fluke eggs in the dung. Animals treated with flukicides had significantly fewer faecal samples positive for eggs (P<0.006) and coproantigen (P<0.05) following treatment compared with the controls. Despite differences in the efficacy profiles among the flukicide-treated groups, there were no significant differences (P>0.05) in growth rates among any of the four treatment groups. There was, however, a significant negative association (P<0.001) between fluke positivity at housing and subsequent growth performance, irrespective of treatment group.

  4. Glycans from Fasciola hepatica Modulate the Host Immune Response and TLR-Induced Maturation of Dendritic Cells

    PubMed Central

    Rodríguez, Ernesto; Noya, Verónica; Cervi, Laura; Chiribao, María Laura; Brossard, Natalie; Chiale, Carolina; Carmona, Carlos; Giacomini, Cecilia; Freire, Teresa

    2015-01-01

    Helminths express various carbohydrate-containing glycoconjugates on their surface, and they release glycan-rich excretion/secretion products that can be very important in their life cycles, infection and pathology. Recent evidence suggests that parasite glycoconjugates could play a role in the evasion of the immune response, leading to a modified Th2-polarized immune response that favors parasite survival in the host. Nevertheless, there is limited information about the nature or function of glycans produced by the trematode Fasciola hepatica, the causative agent of fasciolosis. In this paper, we investigate whether glycosylated molecules from F. hepatica participate in the modulation of host immunity. We also focus on dendritic cells, since they are an important target of immune-modulation by helminths, affecting their activity or function. Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production. During infection, this parasite is able to induce a semi-mature phenotype of DCs expressing low levels of MHCII and secrete IL-10. Furthermore, we show that parasite glycoconjugates mediate the modulation of LPS-induced maturation of DCs since their oxidation restores the capacity of LPS-treated DCs to secrete high levels of the pro-inflammatory cytokines IL-6 and IL-12/23p40 and low levels of the anti-inflammatory cytokine IL-10. Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2. The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis. PMID:26720149

  5. Evaluation of hepatic damage and local immune response in goats immunized with native glutathione S-transferase of Fasciola hepatica.

    PubMed

    Zafra, R; Pérez-Ecija, R A; Buffoni, L; Mendes, R E; Martínez-Moreno, A; Martínez-Moreno, F J; Galisteo, M E Martínez; Pérez, J

    2010-01-01

    Worm burden, hepatic damage and local cellular and humoral immune responses were assessed in goats immunized with glutathione-S-transferase and challenged with Fasciola hepatica. Infected but unimmunized and uninfected control groups were also studied. Hepatic damage was evaluated grossly and microscopically. Local immune response was evaluated by (1) microscopical examination of hepatic lymph nodes (HLNs); (2) analysis of the distribution of CD2(+), CD4(+), CD8(+), T-cell receptor gammadelta(+) lymphocytes and immunoglobulin (Ig) G(+) plasma cells; and (3) investigation of the distribution of cells expressing interleukin (IL)-4 and interferon (IFN)-gamma in the hepatic inflammatory infiltrates and HLNs. Immunized animals did not have significant reduction in fluke number, but there was significant (P<0.05) reduction of fluke size relative to the control groups. The lesions in the two infected groups were similar and consisted of fibrous perihepatitis and white tortuous tracts, mainly involving the left hepatic lobe. Microscopical lesions were similar in both infected groups and were typical of chronic fascioliosis. These included portal fibrosis, inflammatory infiltration with plasma cells, formation of lymphoid follicles, accumulation of haemosiderin-laden macrophages and granulomatous foci. Both infected groups had a marked local immune response characterized by infiltration of CD2(+), CD4(+) and CD8(+) T lymphocytes, and IgG(+) plasma cells in hepatic lesions and in HLNs. There was no expression of IL-4 or INF-gamma by cells in the hepatic inflammatory infiltrate, but expression of INF-gamma in HLNs was much lower than that of IL-4, suggesting an immune response dominated by T helper 2 cells.

  6. Development and application of a fecal antigen diagnostic sandwich ELISA for estimating prevalence of Fasciola gigantica in cattle in central Java, Indonesia.

    PubMed

    Estuningsih, Endah; Spithill, Terry; Raadsma, Herman; Law, Ruby; Adiwinata, G; Meeusen, Els; Piedrafita, David

    2009-04-01

    The purpose of this study was to compare the sensitivity and specificity of an ELISA test to detect Fasciola gigantica antigens (coproantigens) in bovine feces, with fecal egg counting and an ELISA for detecting anti-F. gigantica antibodies in serum. Monoclonal antibodies to cathepsin L were generated and used to capture this antigen in feces of infected cattle. Blood, feces, and livers were collected from 150 cattle at an abattoir in Jakarta, Indonesia, for anti-Fasciola antibodies, coproantigen detection, and F. gigantica egg and worm counts. Fluke recovery varied from 1 to 426 per host, with a mean of 32 flukes. The results showed that the sensitivity and specificity of coproantigen detecting ELISA (95 and 91%, respectively) was better than the anti-F. gigantica antibody ELISA (91 and 88%, respectively) and to fecal egg counting (87 and 100%, respectively). The coproantigen ELISA was able to detect 100% of the cattle with >15 flukes. A survey of 305 cattle in central Java over a 10-mo period validated this test in the field, demonstrating a high prevalence of fascioliasis and establishing the test as a useful diagnostic method to determine patent F. gigantica infections in cattle.

  7. Diagnostic potential of Fasciola gigantica-derived 14.5 kDa fatty acid binding protein in the immunodiagnosis of bubaline fascioliasis.

    PubMed

    Allam, G; Bauomy, I R; Hemyeda, Z M; Diab, T M; Sakran, T F

    2013-06-01

    The 14.5 kDa fatty acid binding protein (FABP) was isolated from the crude extract of adult Fasciola gigantica worms. Polyclonal anti-FABP IgG was generated in rabbits immunized with prepared FABP antigen. Sandwich enzyme-linked immunosorbent assay (ELISA) was applied to detect coproantigen in stools and circulating Fasciola antigen (CA) in sera of 126 water buffaloes by using purified and horseradish peroxidase (HRP)-conjugated anti-FABP IgG. Sandwich ELISA sensitivity was 96.97% and 94.95%; while specificity was 94.12% and 82.35% for coproantigen and CA detection, respectively. However, sensitivity and specificity of the Kato-Katz technique was 73.74% and 100%, respectively. The diagnostic efficacy of sandwich ELISA was 96.55% and 93.1% for coproantigen and CA detection, respectively. In contrast, the diagnostic efficacy of the Kato-Katz technique was 77.59%. In conclusion, these results demonstrate that the purified 14.5 kDa FABP provides a more suitable antigen for immunodiagnosis of early and current bubaline fascioliasis by using sandwich ELISA.

  8. A geographic information system on the potential distribution and abundance of Fasciola hepatica and F. gigantica in east Africa based on Food and Agriculture Organization databases.

    PubMed

    Malone, J B; Gommes, R; Hansen, J; Yilma, J M; Slingenberg, J; Snijders, F; Nachtergaele, F; Ataman, E

    1998-07-31

    An adaptation of a previously developed climate forecast computer model and digital agroecologic database resources available from FAO for developing countries were used to develop a geographic information system risk assessment model for fasciolosis in East Africa, a region where both F. hepatica and F. gigantica occur as a cause of major economic losses in livestock. Regional F. hepatica and F. gigantica forecast index maps were created. Results were compared to environmental data parameters, known life cycle micro-environment requirements and to available Fasciola prevalence survey data and distribution patterns reported in the literature for each species (F. hepatica above 1200 m elevation, F. gigantica below 1800 m, both at 1200-1800 m). The greatest risk, for both species, occurred in areas of extended high annual rainfall associated with high soil moisture and surplus water, with risk diminishing in areas of shorter wet season and/or lower temperatures. Arid areas were generally unsuitable (except where irrigation, water bodies or floods occur) due to soil moisture deficit and/or, in the case of F. hepatica, high average annual mean temperature >23 degrees C. Regions in the highlands of Ethiopia and Kenya were identified as unsuitable for F. gigantica due to inadequate thermal regime, below the 600 growing degree days required for completion of the life cycle in a single year. The combined forecast index (F. hepatica+F. gigantica) was significantly correlated to prevalence data available for 260 of the 1220 agroecologic crop production system zones (CPSZ) and to average monthly normalized difference vegetation index (NDVI) values derived from the advanced very high resolution radiometer (AVHRR) sensor on board the NOAA polar-orbiting satellites. For use in Fasciola control programs, results indicate that monthly forecast parameters, developed in a GIS with digital agroecologic zone databases and monthly climate databases, can be used to define the

  9. Tegument Glycoproteins and Cathepsins of Newly Excysted Juvenile Fasciola hepatica Carry Mannosidic and Paucimannosidic N-glycans

    PubMed Central

    Garcia-Campos, Andres; Cwiklinski, Krystyna; Dalton, John P.; Hokke, Cornelis H.; O’Neill, Sandra; Mulcahy, Grace

    2016-01-01

    Recently, the prevalence of Fasciola hepatica in some areas has increased considerably and the availability of a vaccine to protect livestock from infection would represent a major advance in tools available for controlling this disease. To date, most vaccine-target discovery research on this parasite has concentrated on proteomic and transcriptomic approaches whereas little work has been carried out on glycosylation. As the F. hepatica tegument (Teg) may contain glycans potentially relevant to vaccine development and the Newly Excysted Juvenile (NEJ) is the first lifecycle stage in contact with the definitive host, our work has focused on assessing the glycosylation of the NEJTeg and identifying the NEJTeg glycoprotein repertoire. After in vitro excystation, NEJ were fixed and NEJTeg was extracted. Matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) analysis of released N-glycans revealed that oligomannose and core-fucosylated truncated N-glycans were the most dominant glycan types. By lectin binding studies these glycans were identified mainly on the NEJ surface, together with the oral and ventral suckers. NEJTeg glycoproteins were affinity purified after targeted biotinylation of the glycans and identified using liquid chromatography and tandem mass spectrometry (LC-MS/MS). From the total set of proteins previously identified in NEJTeg, eighteen were also detected in the glycosylated fraction, including the F. hepatica Cathepsin B3 (FhCB3) and two of the Cathepsin L3 (FhCL3) proteins, among others. To confirm glycosylation of cathepsins, analysis at the glycopeptide level by LC-ESI-ion-trap-MS/MS with collision-induced dissociation (CID) and electron-transfer dissociation (ETD) was carried out. We established that cathepsin B1 (FhCB1) on position N80, and FhCL3 (BN1106_s10139B000014, scaffold10139) on position N153, carry unusual paucimannosidic Man2GlcNAc2 glycans. To our knowledge, this is the first description of F

  10. Proteases in egg, miracidium and adult of Fasciola gigantica. Characterization of serine and cysteine proteases from adult.

    PubMed

    Mohamed, Saleh A; Fahmy, Afaf S; Mohamed, Tarek M; Hamdy, Soha M

    2005-10-01

    Proteolytic activity of 0-12 day old eggs, miracidium and adult worm of Fasciola gigantica was assessed and proteases were partially purified by DEAE-Sepharose and CM-cellulose columns. Four forms of protease were separated, PIa, PIb, PIc and PII. Purifications were completed for PIc and PII using Sephacryl S-200 chromatography. A number of natural and synthetic proteins were tested as substrates for F. gigantica PIc and PII. The two proteases had moderate activity levels toward azoalbumin and casein compared to azocasein, while gelatin, hemoglobin, albumin and fibrin had very low affinity toward the two enzymes. Amidolytic substrates are more specific to protease activity. PIc had higher affinity toward BAPNA-HCl (N-benzoyl-arginine-p-nitroanilide-HCl) and BTPNA-HCl (N-benzoyl-tyrosine-p-nitroanilide-HCl) at pH 8.0 indicating that the enzyme was a serine protease. However, PII had higher affinity toward BAPNA at pH 6.5 in the presence of sulfhydryl groups (beta-mercaptoethanol) indicating that the enzyme was a cysteine protease. The effect of specific protease inhibitors on these enzymes was studied. The results confirmed that proteases PIc and PII could be serine and cysteine proteases, respectively. The molecular weights of F. gigantica PIc and PII were 60,000 and 25,000, respectively. F. gigantica PIc and PII had pH optima at 7.5 and 5.5 and K(M) of 2 and 5 mg azocasein/mL, respectively. For amidolytic substrates, PIc had K(M) of 0.3 mM BAPNA/mL and 0.5 mM BTPNA/mL at pH 8.0 and PII had K(M) of 0.6 mM BAPNA/mL at pH 6.5 with reducing agent. F. gigantica PIc and PII had the same optimum temperature at 50 degrees C and were stable up to 40 degrees C. All examined metal cations tested had inhibitory effects toward the two enzymes. From substrate specificity and protease inhibitor studies, PIc and PII could be designated as serine PIc and cysteine PII, respectively. PMID:16102991

  11. Tegument Glycoproteins and Cathepsins of Newly Excysted Juvenile Fasciola hepatica Carry Mannosidic and Paucimannosidic N-glycans.

    PubMed

    Garcia-Campos, Andres; Ravidà, Alessandra; Nguyen, D Linh; Cwiklinski, Krystyna; Dalton, John P; Hokke, Cornelis H; O'Neill, Sandra; Mulcahy, Grace

    2016-05-01

    Recently, the prevalence of Fasciola hepatica in some areas has increased considerably and the availability of a vaccine to protect livestock from infection would represent a major advance in tools available for controlling this disease. To date, most vaccine-target discovery research on this parasite has concentrated on proteomic and transcriptomic approaches whereas little work has been carried out on glycosylation. As the F. hepatica tegument (Teg) may contain glycans potentially relevant to vaccine development and the Newly Excysted Juvenile (NEJ) is the first lifecycle stage in contact with the definitive host, our work has focused on assessing the glycosylation of the NEJTeg and identifying the NEJTeg glycoprotein repertoire. After in vitro excystation, NEJ were fixed and NEJTeg was extracted. Matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) analysis of released N-glycans revealed that oligomannose and core-fucosylated truncated N-glycans were the most dominant glycan types. By lectin binding studies these glycans were identified mainly on the NEJ surface, together with the oral and ventral suckers. NEJTeg glycoproteins were affinity purified after targeted biotinylation of the glycans and identified using liquid chromatography and tandem mass spectrometry (LC-MS/MS). From the total set of proteins previously identified in NEJTeg, eighteen were also detected in the glycosylated fraction, including the F. hepatica Cathepsin B3 (FhCB3) and two of the Cathepsin L3 (FhCL3) proteins, among others. To confirm glycosylation of cathepsins, analysis at the glycopeptide level by LC-ESI-ion-trap-MS/MS with collision-induced dissociation (CID) and electron-transfer dissociation (ETD) was carried out. We established that cathepsin B1 (FhCB1) on position N80, and FhCL3 (BN1106_s10139B000014, scaffold10139) on position N153, carry unusual paucimannosidic Man2GlcNAc2 glycans. To our knowledge, this is the first description of F

  12. The Diterpenoid 7-Keto-Sempervirol, Derived from Lycium chinense, Displays Anthelmintic Activity against both Schistosoma mansoni and Fasciola hepatica

    PubMed Central

    Edwards, Jennifer; Brown, Martha; Peak, Emily; Bartholomew, Barbara; Nash, Robert J.; Hoffmann, Karl F.

    2015-01-01

    Background Two platyhelminths of biomedical and commercial significance are Schistosoma mansoni (blood fluke) and Fasciola hepatica (liver fluke). These related trematodes are responsible for the chronic neglected tropical diseases schistosomiasis and fascioliasis, respectively. As no vaccine is currently available for anti-flukicidal immunoprophylaxis, current treatment is mediated by mono-chemical chemotherapy in the form of mass drug administration (MDA) (praziquantel for schistosomiasis) or drenching (triclabendazole for fascioliasis) programmes. This overreliance on single chemotherapeutic classes has dramatically limited the number of novel chemical entities entering anthelmintic drug discovery pipelines, raising significant concerns for the future of sustainable blood and liver fluke control. Methodology/ Principle Findings Here we demonstrate that 7-keto-sempervirol, a diterpenoid isolated from Lycium chinense, has dual anthelmintic activity against related S. mansoni and F. hepatica trematodes. Using a microtiter plate-based helminth fluorescent bioassay (HFB), this activity is specific (Therapeutic index = 4.2, when compared to HepG2 cell lines) and moderately potent (LD50 = 19.1 μM) against S. mansoni schistosomula cultured in vitro. This anti-schistosomula effect translates into activity against both adult male and female schistosomes cultured in vitro where 7-keto-sempervirol negatively affects motility/behaviour, surface architecture (inducing tegumental holes, tubercle swelling and spine loss/shortening), oviposition rates and egg morphology. As assessed by the HFB and microscopic phenotypic scoring matrices, 7-keto-sempervirol also effectively kills in vitro cultured F. hepatica newly excysted juveniles (NEJs, LD50 = 17.7 μM). Scanning electron microscopy (SEM) evaluation of adult F. hepatica liver flukes co-cultured in vitro with 7-keto-sempervirol additionally demonstrates phenotypic abnormalities including breaches in tegumental integrity and

  13. Effects of sodium chloride exposure on ion regulation in larvae (glochidia) of the freshwater mussel Lampsilis fasciola.

    PubMed

    Nogueira, Lygia S; Bianchini, Adalto; Wood, Chris M; Loro, Vania L; Higgins, Sarah; Gillis, Patricia L

    2015-12-01

    The salinization of freshwater can have negative effects on ecosystem health, with heightened effects in salt-sensitive biota such as glochidia, the larvae of freshwater mussels. However, the toxicological mechanism underlying this sensitivity is unknown. Therefore, Lampsilis fasciola glochidia were exposed to NaCl (nominally 0.25 and 1.0 g/L) prepared in reconstituted moderately-hard water (control), as well as to a dilution of that water (1:4) with ultrapure reference water (diluted control). Unidirectional Na(+) influx (measured with (22)Na) was evaluated after 1, 3 and 48 h of exposure. In addition, unidirectional Cl(-) influx (measured with (36)Cl), whole-body ion (Cl(-) and Na(+)) concentrations, and glochidia viability (measured as the ability to close valves) were assessed after 48 h of exposure. Significantly reduced glochidia viability (56%) was observed after exposure to 1.0 g/L NaCl. Na(+) influx was significantly higher in glochidia exposed to both 0.25 and 1.0 g/L NaCl for 1h than in those kept under control conditions. After 3 and 48 h of exposure, differences in Na(+) influx rate between salt-exposed and control glochidia were generally reduced, indicating that larvae may be able to, at least temporarily, recover their ability to regulate Na(+) influx when exposed to elevated NaCl concentration. Compared to the moderately-hard water control, whole-body Na(+) and Cl(-) concentrations were relatively unchanged in glochidia exposed to 0.25 g/L NaCl, but were significantly elevated in glochidia exposed to 1.0 g/L NaCl and the diluted control. While Na(+) influx rate had recovered to the control level after 48 h of exposure to 1.0 g/L NaCl, Cl(-) influx rate remained elevated, being ~7-fold higher than the Na(+) influx rate. These findings suggest that the loss of viability observed when glochidia were exposed to a high NaCl concentration (1.0 g/L) could be caused by ionoregulatory disturbances mainly associated with an elevated Cl(-) influx.

  14. Differences in the compatibility of infection between the liver flukes Fascioloides magna and Fasciola hepatica in a Colombian population of the snail Galba sp.

    PubMed

    Dreyfuss, G; Correa, A C; Djuikwo-Teukeng, F F; Novobilský, A; Höglund, J; Pankrác, J; Kašný, M; Vignoles, P; Hurtrez-Boussès, S; Pointier, J P; Rondelaud, D

    2015-11-01

    Experimental infections of Galba sp. (origin, Colombia) with allopatric isolates of Fasciola hepatica from France or Fascioloides magna from the Czech Republic were carried out during five successive snail generations to determine if this lymnaeid might sustain complete larval development of either parasite. In snails exposed to F. hepatica, 7 of 400 snails harboured several rediae and only two snails contained a small number of free cercariae on day 50 post-exposure. In contrast, the intensity of F. magna infection in Galba sp. progressively increased from the F1 to F5 generations. Spontaneous cercarial shedding of F. magna occurred in 7 of 100 Galba sp. belonging to the F5 generation and the number of shed cercariae did not differ significantly from that noted in control Galba truncatula of French origin. Galba sp. from Colombia can be added to the list of potential intermediate hosts for F. magna.

  15. Fasciola hepatica phenotypic characterization in Andean human endemic areas: valley versus altiplanic patterns analysed in liver flukes from sheep from Cajamarca and Mantaro, Peru.

    PubMed

    Valero, M Adela; Perez-Crespo, Ignácio; Khoubbane, Messaoud; Artigas, Patricio; Panova, Miroslava; Ortiz, Pedro; Maco, Vicente; Espinoza, José R; Mas-Coma, Santiago

    2012-03-01

    Fascioliasis is a zoonotic parasitic disease caused by Fasciola hepatica and Fasciola gigantica. Of both species, F. hepatica is the only one described in the Americas, mainly transmitted by lymnaeid snail vectors of the Galba/Fossaria group. Human fascioliasis endemic areas are mainly located in high altitude areas of Andean countries. Given the necessity to characterize F. hepatica populations involved, the phenotypic features of fasciolid adults infecting sheep present in human fascioliasis endemic areas were analysed in the Cajamarca Valley and Mantaro Valley (valley transmission patterns) and the northern Bolivian Altiplano (altiplanic transmission pattern). A computer image analysis system (CIAS) was applied on the basis of standardized measurements. The aforementioned highland populations were compared to standard lowland natural and experimental populations of European origin. Liver fluke size was studied by multivariate analyses. Two phenotypic patterns could be distinguished in F. hepatica adult size: the valley pattern (Cajamarca and Mantaro, Peru) and the altiplanic pattern (northern Altiplano, Bolivia). Results showed that the Andean valley population and European standard populations presented a phenotypic homogeneity. The Altiplano population showed a large size range with a pronouncedly lower minimum size indicating that uterus gravidity is reached at a smaller size than in valley populations. The results of this study demonstrate that there is no apparent relationship between the shape of fasciolid adults with regard to altitudinal difference or geographical origin and that allometry-free shape appears as a more stable trait than size in fasciolid species. Results are analysed in terms of intensity/crowding effect aspects and permanent/seasonal transmission characteristics.

  16. Immunodiagnosis of Fasciola gigantica Infection Using Monoclonal Antibody-Based Sandwich ELISA and Immunochromatographic Assay for Detection of Circulating Cathepsin L1 Protease

    PubMed Central

    Anuracpreeda, Panat; Chawengkirttikul, Runglawan; Sobhon, Prasert

    2016-01-01

    Background Tropical fasciolosis caused by Fasciola gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. Parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. Methods In this study, we have produced a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1 (rFgCatL1), and developed both sandwich enzyme-linked immunosorbent assay (sandwich ELISA) and immunochromatographic (IC) test for rapid detection of circulating cathepsin L1 protease (CatL1) in the sera from mice experimentally and cattle naturally infected with Fasciola gigantica. MoAb 4E3 and biotinylated rabbit anti-recombinant CatL1 antibody were selected due to their high reactivities and specificities. Results The lower detection limits of sandwich ELISA and IC test were 3 pg/ml and 0.256 ng/ml, respectively. Sandwich ELISA and IC test could detect F. gigantica infection from day 1 to 35 post infection. In experimental mice, the sensitivity, specificity and accuracy were 95%, 100% and 98.6% (for sandwich ELISA), and 93%, 100% and 98.2% (for IC test), while in natural cattle they were 98.3%, 100% and 99.5% (for sandwich ELISA), and 96.7%, 100% and 99.1% (for IC test). Conclusions These two assay methods showed high efficiencies and precisions for diagnosis of fasciolosis by F. gigantica. PMID:26731402

  17. Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica: expansion of a repertoire of virulence-associated factors.

    PubMed

    Robinson, Mark W; Tort, Jose F; Lowther, Jonathan; Donnelly, Sheila M; Wong, Emily; Xu, Weibo; Stack, Colin M; Padula, Matthew; Herbert, Ben; Dalton, John P

    2008-06-01

    Cathepsin L proteases secreted by the helminth pathogen Fasciola hepatica have functions in parasite virulence including tissue invasion and suppression of host immune responses. Using proteomics methods alongside phylogenetic studies we characterized the profile of cathepsin L proteases secreted by adult F. hepatica and hence identified those involved in host-pathogen interaction. Phylogenetic analyses showed that the Fasciola cathepsin L gene family expanded by a series of gene duplications followed by divergence that gave rise to three clades associated with mature adult worms (Clades 1, 2, and 5) and two clades specific to infective juvenile stages (Clades 3 and 4). Consistent with these observations our proteomics studies identified representatives from Clades 1, 2, and 5 but not from Clades 3 and 4 in adult F. hepatica secretory products. Clades 1 and 2 account for 67.39 and 27.63% of total secreted cathepsin Ls, respectively, suggesting that their expansion was positively driven and that these proteases are most critical for parasite survival and adaptation. Sequence comparison studies revealed that the expansion of cathepsin Ls by gene duplication was followed by residue changes in the S2 pocket of the active site. Our biochemical studies showed that these changes result in alterations in substrate binding and suggested that the divergence of the cathepsin L family produced a repertoire of enzymes with overlapping and complementary substrate specificities that could cleave host macromolecules more efficiently. Although the cathepsin Ls are produced as zymogens containing a prosegment and mature domain, all secreted enzymes identified by MS were processed to mature active enzymes. The prosegment region was highly conserved between the clades except at the boundary of prosegment and mature enzyme. Despite the lack of conservation at this section, sites for exogenous cleavage by asparaginyl endopeptidases and a Leu-Ser[downward arrow]His motif for

  18. FhCaBP2: a Fasciola hepatica calcium-binding protein with EF-hand and dynein light chain domains.

    PubMed

    Thomas, Charlotte M; Timson, David J

    2015-09-01

    FhCaBP2 is a Fasciola hepatica protein which belongs to a family of helminth calcium-binding proteins which combine an N-terminal domain containing two EF-hand motifs and a C-terminal dynein light chain-like (DLC-like) domain. Its predicted structure showed two globular domains joined by a flexible linker. Recombinant FhCaBP2 interacted reversibly with calcium and manganese ions, but not with magnesium, barium, strontium, copper (II), colbalt (II), iron (II), nickel, lead or potassium ions. Cadmium (II) ions appeared to bind non-site-specifically and destabilize the protein. Interaction with either calcium or magnesium ions results in a conformational change in which the protein's surface becomes more hydrophobic. The EF-hand domain alone was able to interact with calcium and manganese ions; the DLC-like domain was not. Alteration of a residue (Asp-58 to Ala) in the second EF-hand motif in this domain abolished ion-binding activity. This suggests that the second EF-hand is the one responsible for ion-binding. FhCaBP2 homodimerizes and the extent of dimerization was not affected by calcium ions or by the aspartate to alanine substitution in the second EF-hand. The isolated EF-hand and DLC-like domains are both capable of homodimerization. FhCaBP2 interacted with the calmodulin antagonists trifluoperazine, chlorpromazine, thiamylal and W7. Interestingly, while chlorpromazine and thiamylal interacted with the EF-hand domain (as expected), trifluoperazine and W7 bound to the DLC-like domain. Overall, FhCaBP2 has distinct biochemical properties compared with other members of this protein family from Fasciola hepatica, a fact which supports the hypothesis that these proteins have different physiological roles.

  19. Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica: expansion of a repertoire of virulence-associated factors.

    PubMed

    Robinson, Mark W; Tort, Jose F; Lowther, Jonathan; Donnelly, Sheila M; Wong, Emily; Xu, Weibo; Stack, Colin M; Padula, Matthew; Herbert, Ben; Dalton, John P

    2008-06-01

    Cathepsin L proteases secreted by the helminth pathogen Fasciola hepatica have functions in parasite virulence including tissue invasion and suppression of host immune responses. Using proteomics methods alongside phylogenetic studies we characterized the profile of cathepsin L proteases secreted by adult F. hepatica and hence identified those involved in host-pathogen interaction. Phylogenetic analyses showed that the Fasciola cathepsin L gene family expanded by a series of gene duplications followed by divergence that gave rise to three clades associated with mature adult worms (Clades 1, 2, and 5) and two clades specific to infective juvenile stages (Clades 3 and 4). Consistent with these observations our proteomics studies identified representatives from Clades 1, 2, and 5 but not from Clades 3 and 4 in adult F. hepatica secretory products. Clades 1 and 2 account for 67.39 and 27.63% of total secreted cathepsin Ls, respectively, suggesting that their expansion was positively driven and that these proteases are most critical for parasite survival and adaptation. Sequence comparison studies revealed that the expansion of cathepsin Ls by gene duplication was followed by residue changes in the S2 pocket of the active site. Our biochemical studies showed that these changes result in alterations in substrate binding and suggested that the divergence of the cathepsin L family produced a repertoire of enzymes with overlapping and complementary substrate specificities that could cleave host macromolecules more efficiently. Although the cathepsin Ls are produced as zymogens containing a prosegment and mature domain, all secreted enzymes identified by MS were processed to mature active enzymes. The prosegment region was highly conserved between the clades except at the boundary of prosegment and mature enzyme. Despite the lack of conservation at this section, sites for exogenous cleavage by asparaginyl endopeptidases and a Leu-Ser[downward arrow]His motif for

  20. Molecular characterization of Fasciola hepatica and phylogenetic analysis based on mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I and cytochrome oxidase subunit I) genes from the North-East of Iran

    PubMed Central

    Reaghi, Saber; Haghighi, Ali; Harandi, Majid Fasihi; Spotin, Adel; Arzamani, Kourosh; Rouhani, Soheila

    2016-01-01

    Aim: Fascioliasis is one of the most zoonotic diseases with global extension. As the epidemiological distribution of Fasciola may lead to various genetic patterns of the parasite, the aim of this study is to identify Fasciola hepatica based on spermatogenesis, and phylogenetic analysis using mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I [ND1] and cytochrome oxidase subunit I) gene marker. Materials and Methods: In this study, 90 F. hepatica collected from 30 cattle at slaughterhouse located in three different geographical locations in the North-East of Iran were evaluated based on spermatogenetic ability and internal transcribed spacer 1 gene restriction fragment length polymorphism pattern. Genetic diversity and phylogenetic relationship using mtDNA gene marker for the isolates from the North-East of Iran, and other countries were then analyzed. Results: Partial sequences of mtDNA showed eight haplotypes in both genes. The phylogenic analysis using neighbor joining as well as maximum likelihood methods showed similar topologies of trees. Pairwise fixation index between different F. hepatica populations calculated from the nucleotide data set of ND1 gene are statistically significant and show the genetic difference. Conclusion: F. hepatica found in this region of Iran has different genetic structures through the other Fasciola populations in the world. PMID:27733809

  1. Prevalence of Fasciola gigantica infection in slaughtered animals in south-eastern Lake Chad area in relation to husbandry practices and seasonal water levels

    PubMed Central

    2014-01-01

    Background Fasciolosis has been described in sub-Saharan Africa in many accounts, but the latest reports from Chad are from the 1970s. Mobile pastoralists perceive liver parasites as a significant problem and think that proximity to Lake Chad can lead to infection. This study aimed to assess the importance of liver fluke infections in mobile pastoralists’ livestock in the south-eastern Lake Chad region. In 2011, all animals presented at three slaughter slabs near Gredaya in the south-eastern Lake Chad area were examined for infection with Fasciola spp. during routine meat inspections. Results This study included 616 goats, 132 sheep and 130 cattle. The prevalence of adult Fasciola gigantica was 68% (CI 60-76%) in cattle, 12% (CI 10-16%) in goats and 23% (CI 16-30%) in sheep. From all infected animals (n = 200), 53% (n = 106) were classified as lightly infected with 1-10 parasites, 18% (n =36) as moderately infected with 11-100 parasites and 29% (n = 58) as heavily infected with more than 100 parasites per animal. Animals grazing close to the shores of Lake Chad had a much higher risk of infection (prevalence =38%; n = 329) than animals not feeding at the lake (n = 353), with only one goat being positive (prevalence = 0.28%). The ethnic group of the owner was a strong determinant for the risk of infection. Ethnic group likely served as a proxy for husbandry practices. Geospatial distribution showed that animals originating from areas close to the lake were more likely to be infected with F. gigantica than those from more distant areas. Conclusions Livestock belonging to ethnic groups which traditionally stay near surface water, and which were reported to feed near Lake Chad, have a high risk of infection with F. gigantica. Pastoralist perception of fasciolosis as a priority health problem was confirmed. Regular preventive and post-exposure treatment is recommended for animals grazing near the lake. However, further economic analysis is

  2. The MF6p/FhHDM-1 Major Antigen Secreted by the Trematode Parasite Fasciola hepatica Is a Heme-binding Protein*

    PubMed Central

    Martínez-Sernández, Victoria; Mezo, Mercedes; González-Warleta, Marta; Perteguer, María J.; Muiño, Laura; Guitián, Esteban; Gárate, Teresa; Ubeira, Florencio M.

    2014-01-01

    Blood-feeding parasites have developed biochemical mechanisms to control heme intake and detoxification. Here we show that a major antigen secreted by Fasciola hepatica, previously reported as MF6p, of unknown function (gb|CCA61804.1), and as FhHDM-1, considered to be a helminth defense molecule belonging to the family of cathelicidin-like proteins (gb|ADZ24001.1), is in fact a heme-binding protein. The heme-binding nature of the MF6p/FhHDM-1 protein was revealed in two independent experiments: (i) immunopurification of the secreted protein·heme complexes with mAb MF6 and subsequent analysis by C8 reversed-phase HPLC and MS/MS spectrometry and (ii) analysis of the binding ability of the synthetic protein to hemin in vitro. By immunohistochemistry analysis, we have observed that MF6p/FhHDM-1 is produced by parenchymal cells and transported to other tissues (e.g. vitellaria and testis). Interestingly, MF6p/FhHDM-1 is absent both in the intestinal cells and in the lumen of cecum, but it can be released through the tegumental surface to the external medium, where it binds to free heme molecules regurgitated by the parasite after hemoglobin digestion. Proteins that are close analogs of the Fasciola MF6p/FhHDM-1 are present in other trematodes, including Clonorchis, Opistorchis, Paragonimus, Schistosoma, and Dicrocoelium. Using UV-visible spectroscopy and immunoprecipitation techniques, we observed that synthetic MF6p/FhHDM-1 binds to hemin with 1:1 stoichiometry and an apparent Kd of 1.14 × 10−6 m−1. We also demonstrated that formation of synthetic MF6p/FhHDM-1·hemin complexes inhibited hemin degradation by hydrogen peroxide and hemin peroxidase-like activity in vitro. Our results suggest that MF6p/FhHDM-1 may be involved in heme homeostasis in trematodes. PMID:24280214

  3. Comparative infectivity of Fasciola hepatica metacercariae from isolates of the main and secondary reservoir animal host species in the Bolivian Altiplano high human endemic region.

    PubMed

    Valero, M A; Mas-Coma, S

    2000-01-01

    Fascioliasis due to Fasciola hepatica (Linnaeus, 1758) is an endemic disease on the Northern Bolivian Altiplano, where human prevalences and intensities are the highest known, sheep and cattle are the main reservoir hosts, and pigs and donkeys the secondary ones. Investigations were carried out to study the viability of metacercariae experimentally obtained from eggs shed by naturally infected Altiplanic sheep, cattle, pigs and donkeys. A total of 157 Wistar rats were infected with doses of 5, 10, 20 and 150 metacercariae. Metacercariae aged for different number of weeks were used to analyse the influence of age on their viability. The number of worms successfully developed in each rat was established by dissection. Results obtained show that metacercarial infectivity is dependent upon storage time, being lower when metacercariae are older. The maximum longevity is 31 weeks using doses of 20 metacercariae per rat and 48 weeks with 150 metacercariae per rat, although in the latter case only a very low percentage of worms is recovered. Age-related infectivity of metacercariae from Altiplanic F. hepatica does not significantly differ from that of the liver fluke in lowlands of other countries. Concerning the influence of the isolate according to host species, results indicate that metacercarial viabilities of pig and donkey isolates are similar to the viabilities of metacercariae of sheep and cattle isolates. Thus, pig and donkey have a high transmission potential capacity concerning this aspect. This fact is of great importance for the control of human and animal fascioliasis in this highly endemic zone.

  4. A single amino acid substitution in isozyme GST mu in Triclabendazole resistant Fasciola hepatica (Sligo strain) can substantially influence the manifestation of anthelmintic resistance.

    PubMed

    Fernández, V; Estein, S; Ortiz, P; Luchessi, P; Solana, V; Solana, H

    2015-12-01

    The helminth parasite Fasciola hepatica causes fascioliasis in human and domestic ruminants. Economic losses due to this infection are estimated in U$S 2000-3000 million yearly. The most common method of control is the use of anthelmintic drugs. However, there is an increased concern about the growing appearance of F. hepatica resistance to Triclabendazole (TCBZ), an anthelmintic with activity over adult and young flukes. F. hepatica has eight Glutathione S-Transferase (GST) isozymes, which are enzymes involved in the detoxification of a wide range of substrates through chemical conjugation with glutathione. In the present work we identified and characterized the GST mu gene isolated from the TCBZ-susceptible and TCBZ-resistant F. hepatica strains. Total RNA was transcribed into cDNA by reverse transcription and a 657 bp amplicon corresponding to the GST mu gene was obtained. The comparative genetic analysis of the GST mu gene of the TCBZ susceptible strain (Cullompton) and TCBZ resistant strain (Sligo) showed three nucleotide changes and one amino acid change at position 143 in the GST mu isozyme of the TCBZ-resistant strain. These results have potential relevance as they contribute better understand the mechanisms that generate resistance to anthelmintics. PMID:26542261

  5. A single amino acid substitution in isozyme GST mu in Triclabendazole resistant Fasciola hepatica (Sligo strain) can substantially influence the manifestation of anthelmintic resistance.

    PubMed

    Fernández, V; Estein, S; Ortiz, P; Luchessi, P; Solana, V; Solana, H

    2015-12-01

    The helminth parasite Fasciola hepatica causes fascioliasis in human and domestic ruminants. Economic losses due to this infection are estimated in U$S 2000-3000 million yearly. The most common method of control is the use of anthelmintic drugs. However, there is an increased concern about the growing appearance of F. hepatica resistance to Triclabendazole (TCBZ), an anthelmintic with activity over adult and young flukes. F. hepatica has eight Glutathione S-Transferase (GST) isozymes, which are enzymes involved in the detoxification of a wide range of substrates through chemical conjugation with glutathione. In the present work we identified and characterized the GST mu gene isolated from the TCBZ-susceptible and TCBZ-resistant F. hepatica strains. Total RNA was transcribed into cDNA by reverse transcription and a 657 bp amplicon corresponding to the GST mu gene was obtained. The comparative genetic analysis of the GST mu gene of the TCBZ susceptible strain (Cullompton) and TCBZ resistant strain (Sligo) showed three nucleotide changes and one amino acid change at position 143 in the GST mu isozyme of the TCBZ-resistant strain. These results have potential relevance as they contribute better understand the mechanisms that generate resistance to anthelmintics.

  6. The performance of a PCR assay for field studies on the prevalence of Fasciola hepatica infection in Galba truncatula intermediate host snails.

    PubMed

    Kozak, Monika; Wedrychowicz, Halina

    2010-02-26

    The aim of this work was to develop a PCR assay for the detection of F. hepatica in Galba truncatula snails and to evaluate its performance in field studies. Primers were designed to amplify a 124bp non-coding tandem repeat found in the Fasciola genome. The result was a banding pattern corresponding to multiples of the initial target sequence. The sensitivity of the PCR was determined on experimentally infected snails. The test was sensitive enough to detect fluke DNA in snails experimentally infected with 1 miracidium, within 12h after exposure. The specificity was determined with Dicrocoelium dendriticum, Paramphistomum cervi and snail DNA. No cross-reactions occurred with DNA of the trematodes or snail DNA. G. truncatula specimens were collected from 4 localities in Eastern Poland, with a total of 192 snails from 12 habitats. The overall prevalence of F. hepatica infection was 26.6% (51/192), ranging from 21.4% to 84.6% in the individual sites. The designed assay was shown to be a valuable epidemiological tool for the purpose of snail infection monitoring. The results on F. hepatica prevalence in snail hosts are the first data from Poland since the 1950s and the only such data based on molecular methods. PMID:19939569

  7. Fasciola hepatica tegumental antigens induce anergic-like T cells via dendritic cells in a mannose receptor-dependent manner.

    PubMed

    Aldridge, Allison; O'Neill, Sandra M

    2016-05-01

    FoxP3(+) Treg cells and anergic T cells are the two regulatory phenotypes of T-cell responses associated with helminth infection. Here, we examine the T-cell responses in mice during Fasciola hepatica infection, and to its tegumental coat antigens (FhTeg) that are shed from the fluke every 2-3 h. FhTeg comprises a rich source of glycoproteins, mainly oligomannose N-glycans that bind to mannose receptor. This study demonstrated a novel mechanism for the T-cell unresponsiveness observed during F. hepatica infection and after injection with FhTeg. Markers of T-cell anergy, such as GRAIL, EGR2, ICOS, and ITCH, are enhanced amongst CD4(+) T-cell populations during infection and following FhTeg injection. This is characterized by a lack of cytokine responses and reduced proliferative activity, which can be reversed with the addition of IL-2. FhTeg-activated dendritic cells (DCs) suppress T cells in vitro as measured by enhanced GRAIL and CTLA4 by RNA and suppressed cytokine expression in anti-CD3 stimulated CD4(+) T cells. FhTeg-treated DCs have enhanced MR expression, which is critical for DC-CD4(+) T-cell communication. Taken together, this study presents markers of anergy in a mouse model of F. hepatica infection, and improves our understanding of host-pathogen interactions and how helminths modulate host immunity. PMID:26931640

  8. Pathological and immunohistochemical study of the liver and hepatic lymph nodes in goats infected with one or more doses of Fasciola hepatica.

    PubMed

    Pérez, J; Martín de las Mulas, J; Carrasco, L; Gutierrez, P N; Martínez-Cruz, M S; Martínez-Moreno, A

    1999-02-01

    Lesions produced by Fasciola hepatica in the liver, gall-bladder and hepatic lymph nodes (HLNs) of four groups of five goats are described; in addition, the distribution of CD3+ T lymphocytes and IgG-lambda light chain-bearing cells was analysed in the hepatic lesions and HLNs. One group of goats received a single oral dose of metacercariae, but the other four groups received four or five doses at different intervals over a period of 11 weeks. Animals that survived were killed 53-55 weeks after the first infective dose. Goats were more susceptible to multiple doses than to a single dose, even when the total number of metacercariae was the same. This greater susceptibility was manifested by the occurrence of deaths and the severity of hepatic lesions. CD3+ lymphocytes were sparse in the infiltrate surrounding the acute migratory tracts, suggesting inhibition of the local cell-mediated immune response. Goats with numerous hepatic calcareous granulomas showed the most severe hepatic damage, including marked cirrhosis, with a striking infiltrate of CD3+ T lymphocytes and lambda IgG- plasma cells, replacing extensive areas of hepatic parenchyma, in which hypertrophy of the smooth endoplasmic reticulum of hepatocytes was evident. These findings were observed mainly in the goats given more than one infective dose.

  9. Isolation of 1E4 IgM Anti-Fasciola hepatica Rediae Monoclonal Antibody from Ascites: Comparison of Two Purification Protocols.

    PubMed

    Alba, Annia; Marcet, Ricardo; Otero, Oscar; Hernández, Hilda M; Figueredo, Mabel; Sarracent, Jorge

    2016-02-01

    Purification of immunoglobulin M (IgM) antibodies could be challenging, and is often characterized by the optimization of the purification protocol to best suit the particular features of the molecule. Here, two different schemes were compared to purify, from ascites, the 1E4 IgM monoclonal antibody (mAb) previously raised against the stage of redia of the trematode Fasciola hepatica. This immunoglobulin is used as capture antibody in an immunoenzymatic assay to detect parasite ongoing infection in its intermediate hosts. The first purification protocol of the 1E4 mAb involved two chromatographic steps: an affinity chromatography on a Concanavalin A matrix followed by size exclusion chromatography. An immunoaffinity chromatography was selected as the second protocol for one-step purification of the antibody using the crude extract of adult parasites coupled to a commercial matrix. Immunoreactivity of the fractions during purification schemes was assessed by indirect immunoenzymatic assays against the crude extract of F. hepatica rediae, while purity was estimated by protein electrophoresis. Losses on the recovery of the antibody isolated by the first purification protocol occurred due to protein precipitation during the concentration of the sample and to low resolution of the size exclusion molecular chromatography step regarding this particular immunoglobulin. The immunoaffinity chromatography using F. hepatica antigens as ligands proved to be the most suitable protocol yielding a pure and immunoreactive antibody. The purification protocols used are discussed regarding efficiency and difficulties. PMID:26828226

  10. Aptitude of Lymnaea palustris and L. stagnalis to Fasciola hepatica larval development through the infection of several successive generations of 4-mm-high snails.

    PubMed

    Vignoles, P; Rondelaud, D; Dreyfuss, G

    2016-06-01

    Bimiracidial infections of Lymnaea palustris and Lymnaea stagnalis (shell height at exposure, 4 mm) with Fasciola hepatica were carried out during six successive snail generations to determine if prevalence and intensity of snail infection increased over time through descendants issuing from eggs laid by parents already exposed to this digenean. Controls were constituted by a French population of Galba truncatula (a single generation) infected according to the same protocol. In a first experiment performed with the F1 to F5 generations of L. palustris, the prevalence and intensity of F. hepatica infection in snails progressively increased. Immature rediae and a few cercariae-containing rediae of the digenean were observed in L. stagnalis from the F3 generation, but no free cercaria was noted in the bodies of this lymnaeid from the F4 to F6 generations. In another experiment carried out with the F6 generation of L. palustris, the prevalence of F. hepatica infection and the number of shed cercariae were significantly lower in L. palustris than in G. truncatula. This mode of snail infection suggests an explanation for cases of human fasciolosis occurring in central France after the collection of wild watercress from beds where L. palustris was the sole lymnaeid.

  11. The T687G SNP in a P-glycoprotein gene of Fasciola hepatica is not associated with resistance to triclabendazole in two resistant Australian populations.

    PubMed

    Elliott, Timothy P; Spithill, Terry W

    2014-11-01

    Triclabendazole (TCBZ) is widely used for control of Fasciola hepatica (liver fluke) in animals and humans and resistance to this drug is now widespread. However, the mechanism of resistance to TCBZ is not known. A T687G single nucleotide polymorphism (SNP) in a P-glycoprotein gene was proposed as a molecular marker for TCBZ resistance in F. hepatica (Wilkinson et al., 2012). We analyzed this Pgp gene from TCBZ-susceptible and TCBZ-resistant populations from Australia to determine if the SNP was a marker for TCBZ resistance. From the 21 parasites studied we observed 27 individual haplotypes in the Pgp sequences which comprised seven haplotypic groups (A-G), with haplotypes A and B representing 81% of the total observed. The T687G SNP was not observed in either of the resistant or susceptible populations. We conclude that the T687G SNP in this Pgp gene is not associated with TCBZ resistance in these Australian F. hepatica populations and therefore unlikely to be a universal molecular marker for TCBZ resistance.

  12. Natural prevalence in Cuban populations of the lymnaeid snail Galba cubensis infected with the liver fluke Fasciola hepatica: small values do matter.

    PubMed

    Vázquez, Antonio A; Sánchez, Jorge; Alba, Annia; Pointier, Jean-Pierre; Hurtrez-Boussès, Sylvie

    2015-11-01

    Natural infections of lymnaeid snails by Fasciola hepatica are of primary importance to study transmission. Also, infected snails in the field can be used to explore the existing compatibility in host-parasite interactions. This paper aimed to describe the infection rate of Galba cubensis populations in fasciolosis transmission areas. Eight sites were sampled in western Cuba and 24 infected snails at six sites were found. The mean prevalence was 2.94% and the maximum value was 11.4%. The intensity of parasite infection was assessed as the number of rediae inside a single snail. High variation within the sites examined was observed, but a maximum of 76 rediae was recovered from one individual. Although the presence of two other trematode families (Schistosomatidae and Paramphistomatidae) was discovered in dissected individuals, no co-infection with F. hepatica was observed. This is the first time a study of natural prevalence of F. hepatica infection is carried out in Cuba, considered a hyper endemic country for bovine fasciolosis. Our results suggest that fasciolosis transmission may occur even when the number of infected snails remains relatively low.

  13. Activity of cholinesterases, pyruvate kinase and adenosine deaminase in rats experimentally infected by Fasciola hepatica: Influences of these enzymes on inflammatory response and pathological findings.

    PubMed

    Baldissera, Matheus D; Bottari, Nathieli B; Mendes, Ricardo E; Schwertz, Claiton I; Lucca, Neuber J; Dalenogare, Diessica; Bochi, Guilherme V; Moresco, Rafael N; Morsch, Vera M; Schetinger, Maria R C; Rech, Virginia C; Jaques, Jeandre A; Da Silva, Aleksandro S

    2015-11-01

    The aim of this study was to investigate acetylcholinesterase (AChE) in total blood and liver tissue; butyrylcholinesterase (BChE) in serum and liver tissue; adenosine deaminase (ADA) in serum and liver tissue; and pyruvate kinase (PK) in liver tissue of rats experimentally infected by Fasciola hepatica. Animals were divided into two groups with 12 animals each, as follows: group A (uninfected) and group B (infected). Samples were collected at 20 (A1 and B1;n=6 each) and 150 (A2 and B2; n=6 each) days post-infection (PI). Infected animals showed an increase in AChE activity in whole blood and a decrease in AChE activity in liver homogenates (P<0.05) at 20 and 150 days PI. BChE and PK activities were decreased (P<0.05) in serum and liver homogenates of infected animals at 150 days PI. ADA activity was decreased in serum at 20 and 150 days PI, while in liver homogenates it was only decreased at 150 days PI (P<0.05). Aspartate aminotransferase and alanine aminotransferase activities in serum were increased (P<0.05), while concentrations of total protein and albumin were decreased (P<0.05) when compared to control. The histological analysis revealed fibrous perihepatitis and necrosis. Therefore, we conclude that the liver fluke is associated with cholinergic and purinergic dysfunctions, which in turn may influence the pathogenesis of the disease.

  14. Fasciola hepatica: a light and electron microscope study of the ovary and of the development of oocytes within eggs in the uterus provides an insight into reproductive strategy.

    PubMed

    Hanna, R E B; Moffett, D; Forster, F I; Trudgett, A G; Brennan, G P; Fairweather, I

    2016-05-15

    The ultrastructure of the ovary of Fasciola hepatica collected from field-infected sheep, was compared with that of flukes from laboratory-infected rats harbouring the Oberon or the Cullompton fluke isolate. At the periphery of the ovarian tubules, in all flukes, interstitial tissue was identified that appears to provide physical support and facilitate the metabolism of the germinal-line cells. Oogonia undergo mitotic division to maintain the cell population and to produce oocytes. Early oocytes feature conspicuous synaptonemal complexes in the nucleoplasm, and these become less evident as the oocytes grow in size, move towards the core of the ovarian tubule, and synthesise osmiophilic bodies. The latter may represent cortical granules, and serve to block polyspermy. The identity of the synaptonemal complexes was confirmed by immunocytochemical labelling of synaptonemal proteins. The occurrence of synaptonemal complexes in the oocytes of all fluke types examined indicates that pairing of bivalent chromosomes, with the potential for genetic recombination and chiasmata formation, is a feature of the triploid aspermic parthenogenetic Cullompton flukes, as well as of the wild-type out-breeding field-derived and Oberon isolate flukes. In oocytes within shelled eggs in the proximal uterus of all flukes, condensed chromosomes align at meiotic metaphase plates. Following the reduction division, two equal pronuclei appear in each oocyte in the distal uterus. On the basis of these observations, a mechanism of facultative parthenogenesis for F. hepatica is proposed that accommodates the survival and clonal expansion of triploid aspermic isolates.

  15. Standardisation of egg-viability assays for Fasciola hepatica and Calicophoron daubneyi: A tool for evaluating new technologies of parasite control.

    PubMed

    Chryssafidis, Andreas Lazaros; Fu, Yan; De Waal, Theo; Mulcahy, Grace

    2015-05-30

    Fasciola hepatica and Calicophoron daubneyi, liver and rumen flukes respectively, infect ruminants throughout Europe. There is considerable interest in the development of vaccines and in testing new potential anthelmintic agents against these species. One potential target of new control measures is the parasite egg, as interference at this stage of the life cycle could aid in blocking the transmission of infection, and some experimental vaccines have been shown to affect egg viability. In this study, we describe the standardisation of protocols to evaluate the viability of eggs of these two parasites. Eggs were recovered from adult parasites collected in a commercial abattoir, from naturally infected cattle. A protocol for in vitro development of F. hepatica eggs was optimised based on previously published methods, with variations in duration and temperature of incubation. A new protocol for measurement of rumen fluke egg development in vitro was designed, based on testing different temperatures and periods of incubation, with or without light exposure. The protocols described here may be used in the future for comparing experimental groups when new technologies for parasite control are tested. In addition, the methods described for C. daubneyi present new information on the biology of this parasite.

  16. The Effect of an Eextremely Low Frequency Magnetic Field on Larvae Production in the Parasite-Host System: Fasciola hepatica-Galba truncatula: a Preliminary Study.

    PubMed

    Kołodziejczyk, Lidia; Podraza, Wojciech; Gonet, Bolesław; Dzika, Ewa; Kosik-Bogacka, Danuta I

    2016-01-01

    The aim of this study was to determine the effect of an extremely low-frequency magnetic field (ELFMF) on the production of liver fluke larvae in a parasite-host system: Fasciola hepatica--Galba truncatula. Both F. hepatica eggs and F. hepatica-infected snails were exposed to an ELFMF (50 Hz, 2.0 mT) for 14 days and 36 days, respectively. F. hepatica-infected snails were divided into 4 groups, 10 specimens each. The snails of groups I and II were infected with F. hepatica larvae--miracidia obtained from control cultures, while the snails of groups III and IV were infected with miracidia reared from eggs that had been incubated in an ELFMF. After infection, the snails of groups II and IV were placed in an ELFMF, while those of groups I (control) and III were housed outside the ELFMF. At 36 days post-infection (dpi) there were no statistically significant differences between the number of F. hepatica larvae--cercariae and metacercariae, obtained from G. truncatula snails in the control group (group I) and the snail groups exposed to ELFMF (groups II, III and IV). However, a statistically significant difference between the average number of F. hepatica larvae in snail groups III and IV may indicate that the duration of exposure to ELFMF, i.e. embryogenesis period vs. the entire larval development, played a role in the production of F. hepatica larvae, and resulted in a reduction of their number.

  17. The Effect of an Eextremely Low Frequency Magnetic Field on Larvae Production in the Parasite-Host System: Fasciola hepatica-Galba truncatula: a Preliminary Study.

    PubMed

    Kołodziejczyk, Lidia; Podraza, Wojciech; Gonet, Bolesław; Dzika, Ewa; Kosik-Bogacka, Danuta I

    2016-01-01

    The aim of this study was to determine the effect of an extremely low-frequency magnetic field (ELFMF) on the production of liver fluke larvae in a parasite-host system: Fasciola hepatica--Galba truncatula. Both F. hepatica eggs and F. hepatica-infected snails were exposed to an ELFMF (50 Hz, 2.0 mT) for 14 days and 36 days, respectively. F. hepatica-infected snails were divided into 4 groups, 10 specimens each. The snails of groups I and II were infected with F. hepatica larvae--miracidia obtained from control cultures, while the snails of groups III and IV were infected with miracidia reared from eggs that had been incubated in an ELFMF. After infection, the snails of groups II and IV were placed in an ELFMF, while those of groups I (control) and III were housed outside the ELFMF. At 36 days post-infection (dpi) there were no statistically significant differences between the number of F. hepatica larvae--cercariae and metacercariae, obtained from G. truncatula snails in the control group (group I) and the snail groups exposed to ELFMF (groups II, III and IV). However, a statistically significant difference between the average number of F. hepatica larvae in snail groups III and IV may indicate that the duration of exposure to ELFMF, i.e. embryogenesis period vs. the entire larval development, played a role in the production of F. hepatica larvae, and resulted in a reduction of their number. PMID:27172713

  18. Cloning and Expression of the Major Secreted Cathepsin B-Like Protein from Juvenile Fasciola hepatica and Analysis of Immunogenicity following Liver Fluke Infection

    PubMed Central

    Law, Ruby H. P.; Smooker, Peter M.; Irving, James A.; Piedrafita, David; Ponting, Rebecca; Kennedy, Nicholas J.; Whisstock, James C.; Pike, Robert N.; Spithill, Terry W.

    2003-01-01

    The functions of the cathepsin B-like proteases in liver flukes are unknown and analysis has been hindered by a lack of protein for study, since the protein is produced in small amounts by juvenile flukes. To circumvent this, we isolated and characterized a cDNA encoding the major secreted cathepsin B from Fasciola hepatica. The predicted preproprotein is 339 amino acids in length, with the mature protease predicted to be 254 amino acids long, and shows significant similarity to parasite and mammalian cathepsin B. Only one of the two conserved histidine residues required for cathepsin B exopeptidase activity is predicted to be present. Recombinant preproprotein was produced in yeast, and it was shown that the recombinant proprotein can undergo a degree of self-processing in vitro to the mature form, which is active against gelatin and synthetic peptide substrates. The recombinant protein is antigenic in vaccinated rats, and antibodies to the protein are detected early after infection of rats and sheep with F. hepatica. The kinetics of the response to cathepsin B and cathepsin L after infection of sheep and rats confirm the temporal expression of these proteins during the life cycle of the parasite. PMID:14638781

  19. The population density of Lymnaea columella (Say, 1817) (Mollusca, Lymnaeidae) an intermediate host of Fasciola hepatica (Linnaeus, 1758), in the Caparaó microregion, ES, Brazil.

    PubMed

    D'Almeida, S C G; Freitas, D F; Carneiro, M B; Camargo, P F; Azevedo, J C; Martins, I V F

    2016-06-01

    The aim of this study was to monitor the population density of Lymnaea columella, an intermediate host of Fasciola hepatica, in various aquatic habitats and in drinking water in the area of the Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, on Caparaó Microregion, municipality of Alegre, state of Espírito Santo, Brazil. Monthly samplings were performed at certain points between drainage areas and drinking water in cattle and goat production systems during the years 2010 to 2013. The mean temperature, precipitation and the frequency of samples of L. columella were analysed graphically according the monthly average during the study period. A total of 2,038 molluscs were collected, 1558 of which were L. columella, that predominated in all sampled points. The highest average of specimens observed for L. columella was in the years 2010 and 2013 (51.0), and occurred decreased in 2011 (19.8). The temperature and precipitation averaged is 23.7 °C and 141 mm/year, respectively. Rainfall peak occurred in March (2011, 2013) and November (2012), during these periods the population of L. columella growth. There was no significant difference in the relationship between the specimens observed with seasons (dry-wet), thus the population of L. columella remained stable and can be found throughout the year.

  20. Fasciola hepatica: a light and electron microscope study of the ovary and of the development of oocytes within eggs in the uterus provides an insight into reproductive strategy.

    PubMed

    Hanna, R E B; Moffett, D; Forster, F I; Trudgett, A G; Brennan, G P; Fairweather, I

    2016-05-15

    The ultrastructure of the ovary of Fasciola hepatica collected from field-infected sheep, was compared with that of flukes from laboratory-infected rats harbouring the Oberon or the Cullompton fluke isolate. At the periphery of the ovarian tubules, in all flukes, interstitial tissue was identified that appears to provide physical support and facilitate the metabolism of the germinal-line cells. Oogonia undergo mitotic division to maintain the cell population and to produce oocytes. Early oocytes feature conspicuous synaptonemal complexes in the nucleoplasm, and these become less evident as the oocytes grow in size, move towards the core of the ovarian tubule, and synthesise osmiophilic bodies. The latter may represent cortical granules, and serve to block polyspermy. The identity of the synaptonemal complexes was confirmed by immunocytochemical labelling of synaptonemal proteins. The occurrence of synaptonemal complexes in the oocytes of all fluke types examined indicates that pairing of bivalent chromosomes, with the potential for genetic recombination and chiasmata formation, is a feature of the triploid aspermic parthenogenetic Cullompton flukes, as well as of the wild-type out-breeding field-derived and Oberon isolate flukes. In oocytes within shelled eggs in the proximal uterus of all flukes, condensed chromosomes align at meiotic metaphase plates. Following the reduction division, two equal pronuclei appear in each oocyte in the distal uterus. On the basis of these observations, a mechanism of facultative parthenogenesis for F. hepatica is proposed that accommodates the survival and clonal expansion of triploid aspermic isolates. PMID:27084479

  1. Isolation of 1E4 IgM Anti-Fasciola hepatica Rediae Monoclonal Antibody from Ascites: Comparison of Two Purification Protocols.

    PubMed

    Alba, Annia; Marcet, Ricardo; Otero, Oscar; Hernández, Hilda M; Figueredo, Mabel; Sarracent, Jorge

    2016-02-01

    Purification of immunoglobulin M (IgM) antibodies could be challenging, and is often characterized by the optimization of the purification protocol to best suit the particular features of the molecule. Here, two different schemes were compared to purify, from ascites, the 1E4 IgM monoclonal antibody (mAb) previously raised against the stage of redia of the trematode Fasciola hepatica. This immunoglobulin is used as capture antibody in an immunoenzymatic assay to detect parasite ongoing infection in its intermediate hosts. The first purification protocol of the 1E4 mAb involved two chromatographic steps: an affinity chromatography on a Concanavalin A matrix followed by size exclusion chromatography. An immunoaffinity chromatography was selected as the second protocol for one-step purification of the antibody using the crude extract of adult parasites coupled to a commercial matrix. Immunoreactivity of the fractions during purification schemes was assessed by indirect immunoenzymatic assays against the crude extract of F. hepatica rediae, while purity was estimated by protein electrophoresis. Losses on the recovery of the antibody isolated by the first purification protocol occurred due to protein precipitation during the concentration of the sample and to low resolution of the size exclusion molecular chromatography step regarding this particular immunoglobulin. The immunoaffinity chromatography using F. hepatica antigens as ligands proved to be the most suitable protocol yielding a pure and immunoreactive antibody. The purification protocols used are discussed regarding efficiency and difficulties.

  2. Distribution of Fasciola hepatica and F. gigantica in the endemic area of Guilan, Iran: Relationships between zonal overlap and phenotypic traits.

    PubMed

    Ashrafi, Keyhan; Valero, M Adela; Peixoto, Raquel V; Artigas, Patricio; Panova, Miroslava; Mas-Coma, Santiago

    2015-04-01

    Fascioliasis is a zoonotic disease emerging in numerous parts of the world. In any endemic area, the characterisation of scenarios and patterns of infection must always be considered the starting point before implementing any control measure. Fascioliasis is a parasitic disease of different epidemiological, pathological and control characteristics depending on the endemic area and the causal agent, Fasciola hepatica and Fasciolagigantica. Classically it has been accepted that F. hepatica is present worldwide, while the distribution of the two species overlaps in many areas of Africa and Asia. Fascioliasis caused by F. hepatica, F. gigantica and intermediate forms is present in Guilan province, a complicated epidemiological situation where the highest human infection rates have been described in Iran. Morphometric tools were used to analyse the possible relationship between liver-fluke metric traits and geographical and altitudinal distribution. This is the first study in which a detailed distribution of both Fasciola species is analysed in a human fascioliasis endemic area with a zonal overlap transmission pattern. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected livestock (cattle, buffaloes, sheep and goats). The distribution of the % F. hepatica-like (F.h.) and F. gigantica-like (F.g.) flukes detected in each liver versus altitude (m) in each group was analysed. The presence of F.g. specimens mainly in locations below sea level (average: 11.23% F.h., 88.77% F.g.), the presence of both species with similar intensity at 1-99m (average: 56.95% F.h., 43.05% F.g.) and the presence of F.h. specimens mainly from 100 to 999m (average: 71.69% F.h., 28.31% F.g.) as well as in locations with an altitude above 1000m (average: 97.48% F.h., 2.52% F.g.) are noteworthy. A significant positive correlation was obtained between altitude and % F.h., and a significant negative correlation was obtained between altitude and

  3. Distribution of Fasciola hepatica and F. gigantica in the endemic area of Guilan, Iran: Relationships between zonal overlap and phenotypic traits.

    PubMed

    Ashrafi, Keyhan; Valero, M Adela; Peixoto, Raquel V; Artigas, Patricio; Panova, Miroslava; Mas-Coma, Santiago

    2015-04-01

    Fascioliasis is a zoonotic disease emerging in numerous parts of the world. In any endemic area, the characterisation of scenarios and patterns of infection must always be considered the starting point before implementing any control measure. Fascioliasis is a parasitic disease of different epidemiological, pathological and control characteristics depending on the endemic area and the causal agent, Fasciola hepatica and Fasciolagigantica. Classically it has been accepted that F. hepatica is present worldwide, while the distribution of the two species overlaps in many areas of Africa and Asia. Fascioliasis caused by F. hepatica, F. gigantica and intermediate forms is present in Guilan province, a complicated epidemiological situation where the highest human infection rates have been described in Iran. Morphometric tools were used to analyse the possible relationship between liver-fluke metric traits and geographical and altitudinal distribution. This is the first study in which a detailed distribution of both Fasciola species is analysed in a human fascioliasis endemic area with a zonal overlap transmission pattern. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected livestock (cattle, buffaloes, sheep and goats). The distribution of the % F. hepatica-like (F.h.) and F. gigantica-like (F.g.) flukes detected in each liver versus altitude (m) in each group was analysed. The presence of F.g. specimens mainly in locations below sea level (average: 11.23% F.h., 88.77% F.g.), the presence of both species with similar intensity at 1-99m (average: 56.95% F.h., 43.05% F.g.) and the presence of F.h. specimens mainly from 100 to 999m (average: 71.69% F.h., 28.31% F.g.) as well as in locations with an altitude above 1000m (average: 97.48% F.h., 2.52% F.g.) are noteworthy. A significant positive correlation was obtained between altitude and % F.h., and a significant negative correlation was obtained between altitude and

  4. Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach☆

    PubMed Central

    Cwiklinski, Krystyna; Allen, Katherine; LaCourse, James; Williams, Diana J.; Paterson, Steve; Hodgkinson, Jane E.

    2015-01-01

    The liver fluke, Fasciola hepatica is an economically important pathogen of sheep and cattle and has been described by the WHO as a re-emerging zoonosis. Control is heavily reliant on the use of drugs, particularly triclabendazole and as a result resistance has now emerged. The population structure of F. hepatica is not well known, yet it can impact on host–parasite interactions and parasite control with drugs, particularly regarding the spread of triclabendazole resistance. We have identified 2448 potential microsatellites from 83 Mb of F. hepatica genome sequence using msatfinder. Thirty-five loci were developed and optimised for microsatellite PCR, resulting in a panel of 15 polymorphic loci, with a range of three to 15 alleles. This panel was validated on genomic DNA from 46 adult F. hepatica; 38 liver flukes sourced from a Northwest abattoir, UK and 8 liver flukes from an established isolate (Shrewsbury; Ridgeway Research). Evidence for null alleles was found at four loci (Fh_1, Fh_8, Fh_13 and Fh_14), which showed markedly higher levels of homozygosity than the remaining 11 loci. Of the 38 liver flukes isolated from cattle livers (n = 10) at the abattoir, 37 genotypes were identified. Using a multiplex approach all 15 loci could be amplified from several life cycle stages that typically yield low amounts of DNA, including metacercariae, the infective life cycle stage present on pasture, highlighting the utility of this multiplex microsatellite panel. This study reports the largest panel of microsatellite markers available to date for population studies of F. hepatica and the first multiplex panel of microsatellite markers that can be used for several life cycle stages. PMID:25796359

  5. Biochemical Characterization and Differential Expression of a 16.5-Kilodalton Tegument-Associated Antigen from the Liver Fluke Fasciola hepatica

    PubMed Central

    Gaudier, José F.; Cabán-Hernández, Kimberly; Osuna, Antonio

    2012-01-01

    A cDNA encoding a 16.5-kDa protein termed FhTP16.5 was identified by immunoscreening of a cDNA library from Fasciola hepatica adult flukes using pooled sera from rabbits infected with F. hepatica for 4 weeks. Quantitative reverse transcriptase PCR (qPCR) analysis revealed that FhTP16.5 is not expressed in unembryonated eggs. It is poorly expressed in miracidia and highly expressed at the juvenile and adult stages; however, significant differences were found between the expression levels of FhTP16.5 in juveniles versus adult flukes. Recombinant FhTP16.5 was expressed at high levels in Escherichia coli, purified by affinity chromatography, and used to raise anti-FhTP16.5 polyclonal antibodies in rabbits. Immunoblot analysis using the anti-FhTP16.5 IgG antibody identified FhTP16.5 in crude and tegumental extracts and in excretory-secretory products of F. hepatica. The protein was not detected in crude extracts of Schistosoma mansoni or Schistosoma japonicum. Antibodies to FhTP16.5 were detected in the sera of rabbits at 3 to 12 weeks of F. hepatica infection as well as in the sera of humans with chronic fascioliasis; these findings suggest that FhTP16.5 could be a good antigen for serodiagnosis of fascioliasis. Immunohistochemistry demonstrated that FhTP16.5 localizes to the surface of the tegument of various developmental stages and in parenchymal tissues of the adult fluke. Such specific localization makes FhTP16.5 an attractive target for immunoprophylaxis or chemotherapy. PMID:22278327

  6. Fasciola hepatica: Histological changes in the somatic and reproductive tissues of liver fluke following closantel treatment of experimentally-infected sheep.

    PubMed

    Scarcella, S; Hanna, R E B; Brennan, G P; Solana, H; Fairweather, I

    2016-01-15

    Lambs infected with the Cullompton isolate of Fasciola hepatica were treated orally or subcutaneously with 10mg/kg of closantel at 16 weeks post-infection. Adult flukes were recovered from the liver of individual animals at 12h, 24h, or 36h post-treatment. The flukes were processed for histological analysis. In general, degenerative changes in the reproductive and somatic tissues were progressive, and were most marked in flukes exposed to closantel in vivo for 36h. However, flukes from a 12h subcutaneously-treated lamb showed marked deterioration of the testis, possibly because a portion of the dose has been delivered intravenously. Fewer intact eggs were seen in the uterus of flukes exposed to closantel for longer times (whether administered subcutaneously or orally to the host). The most conspicuous closantel-induced effect in flukes from treated hosts was progressive damage to the tegumental syncytium. While the flukes from 24h-treated hosts showed relatively minor damage to limited areas of the syncytium, towards the posterior end, the flukes from 36h-treated hosts (and flukes from the lamb that putatively received intravenous dosage) had lost large areas of the surface syncytium from the posterior end and dorsal surface, although the syncytium over the anterior end and the anterior ventral surface was largely spared. In areas where the syncytium had sloughed, the underlying structures such as the vitelline follicles, gut profiles and testis profiles, showed marked degeneration and breakdown. Other changes included cell depletion and early stage apoptosis in the testis, ovary and vitelline follicles. This study establishes a model for histological changes in closantel-sensitive F. hepatica exposed to closantel in vivo. Histopathological studies could be complementary to the efficacy controlled test for for closantel resistance in fluke populations. PMID:26790736

  7. Fasciola hepatica: Histological changes in the somatic and reproductive tissues of liver fluke following closantel treatment of experimentally-infected sheep.

    PubMed

    Scarcella, S; Hanna, R E B; Brennan, G P; Solana, H; Fairweather, I

    2016-01-15

    Lambs infected with the Cullompton isolate of Fasciola hepatica were treated orally or subcutaneously with 10mg/kg of closantel at 16 weeks post-infection. Adult flukes were recovered from the liver of individual animals at 12h, 24h, or 36h post-treatment. The flukes were processed for histological analysis. In general, degenerative changes in the reproductive and somatic tissues were progressive, and were most marked in flukes exposed to closantel in vivo for 36h. However, flukes from a 12h subcutaneously-treated lamb showed marked deterioration of the testis, possibly because a portion of the dose has been delivered intravenously. Fewer intact eggs were seen in the uterus of flukes exposed to closantel for longer times (whether administered subcutaneously or orally to the host). The most conspicuous closantel-induced effect in flukes from treated hosts was progressive damage to the tegumental syncytium. While the flukes from 24h-treated hosts showed relatively minor damage to limited areas of the syncytium, towards the posterior end, the flukes from 36h-treated hosts (and flukes from the lamb that putatively received intravenous dosage) had lost large areas of the surface syncytium from the posterior end and dorsal surface, although the syncytium over the anterior end and the anterior ventral surface was largely spared. In areas where the syncytium had sloughed, the underlying structures such as the vitelline follicles, gut profiles and testis profiles, showed marked degeneration and breakdown. Other changes included cell depletion and early stage apoptosis in the testis, ovary and vitelline follicles. This study establishes a model for histological changes in closantel-sensitive F. hepatica exposed to closantel in vivo. Histopathological studies could be complementary to the efficacy controlled test for for closantel resistance in fluke populations.

  8. Resistance of Fasciola hepatica against Triclabendazole in cattle in Cajamarca (Peru): a clinical trial and an in vivo efficacy test in sheep.

    PubMed

    Ortiz, P; Scarcella, S; Cerna, C; Rosales, C; Cabrera, M; Guzmán, M; Lamenza, P; Solana, H

    2013-07-01

    Fasciolosis caused by Fasciola hepatica, is the most prevalent parasitic disease in dairy cattle from the northern region of Cajamarca, Peru. The control of this parasite is based on the use of Triclabendazole (TCBZ), a drug that has been used for more than fifteen years in this area. Recent studies, however, have reported a lack of clinical efficacy after treating dairy cattle. This research was aimed to determine the efficacy of TCBZ in a clinical trial. Eleven dairy cows all positive to F. hepatica identified by presence of eggs in feces, were treated with TCBZ (Fasinex(®) 10%) at 12 mg/kg body weight. Fourteen and thirty days after treatment, the animals were analyzed for F. hepatica eggs in their feces by the fecal egg count reduction test. The results found show an overall efficacy of 31.05% and 13. 63% (14 and 30 days post treatment, respectively). Furthermore, an in vivo efficacy test was conducted in sheep with metacercariae obtained from eggs isolated from a cow clinically resistant to TCBZ. Eleven sheep divided in two groups, a control group with no treatment (n=5) and a treated group (n=6) were all infected with two hundred metacercariae. One hundred and six days after infection all the animals demonstrated F. hepatica eggs in their feces, confirming the presence of adult parasites in their livers. The animals were then treated with TCBZ (Fasinex(®) 10%) at 10mg/kg body weight. Fifteen days later, the animals were sacrificed and the number of F. hepatica in their livers counted. The results of this experiment showed an efficacy of the flukicide of 25.2% confirming the resistance to TCBZ of the F. hepatica isolated from dairy cattle in Cajamarca, Peru.

  9. Fasciola hepatica Surface Coat Glycoproteins Contain Mannosylated and Phosphorylated N-glycans and Exhibit Immune Modulatory Properties Independent of the Mannose Receptor.

    PubMed

    Ravidà, Alessandra; Aldridge, Allison M; Driessen, Nicole N; Heus, Ferry A H; Hokke, Cornelis H; O'Neill, Sandra M

    2016-04-01

    Fascioliasis, caused by the liver fluke Fasciola hepatica, is a neglected tropical disease infecting over 1 million individuals annually with 17 million people at risk of infection. Like other helminths, F. hepatica employs mechanisms of immune suppression in order to evade its host immune system. In this study the N-glycosylation of F. hepatica's tegumental coat (FhTeg) and its carbohydrate-dependent interactions with bone marrow derived dendritic cells (BMDCs) were investigated. Mass spectrometric analysis demonstrated that FhTeg N-glycans comprised mainly of oligomannose and to a lesser extent truncated and complex type glycans, including a phosphorylated subset. The interaction of FhTeg with the mannose receptor (MR) was investigated. Binding of FhTeg to MR-transfected CHO cells and BMDCs was blocked when pre-incubated with mannan. We further elucidated the role played by MR in the immunomodulatory mechanism of FhTeg and demonstrated that while FhTeg's binding was significantly reduced in BMDCs generated from MR knockout mice, the absence of MR did not alter FhTeg's ability to induce SOCS3 or suppress cytokine secretion from LPS activated BMDCs. A panel of negatively charged monosaccharides (i.e. GlcNAc-4P, Man-6P and GalNAc-4S) were used in an attempt to inhibit the immunoregulatory properties of phosphorylated oligosaccharides. Notably, GalNAc-4S, a known inhibitor of the Cys-domain of MR, efficiently suppressed FhTeg binding to BMDCs and inhibited the expression of suppressor of cytokine signalling (SOCS) 3, a negative regulator the TLR and STAT3 pathway. We conclude that F. hepatica contains high levels of mannose residues and phosphorylated glycoproteins that are crucial in modulating its host's immune system, however the role played by MR appears to be limited to the initial binding event suggesting that other C-type lectin receptors are involved in the immunomodulatory mechanism of FhTeg. PMID:27104959

  10. Invasion biology meets parasitology: a case study of parasite spill-back with Egyptian Fasciola gigantica in the invasive snail Pseudosuccinea columella.

    PubMed

    Grabner, Daniel S; Mohamed, Faten A M M; Nachev, Milen; Méabed, Eman M H; Sabry, Abdel Hameed A; Sures, Bernd

    2014-01-01

    The liver fluke Fasciola gigantica is a trematode parasite of ruminants and humans that occurs naturally in Africa and Asia. Cases of human fascioliasis, attributable at least in part to F. gigantica, are significantly increasing in the last decades. The introduced snail species Galba truncatula was already identified to be an important intermediate host for this parasite and the efficient invader Pseudosuccinea columella is another suspect in this case. Therefore, we investigated snails collected in irrigation canals in Fayoum governorate in Egypt for prevalence of trematodes with focus on P. columella and its role for the transmission of F. gigantica. Species were identified morphologically and by partial sequencing of the cytochrome oxidase subunit I gene (COI). Among all 689 snails found at the 21 sampling sites, P. columella was the most abundant snail with 296 individuals (42.96%) and it was also the most dominant species at 10 sites. It was not found at 8 sites. Molecular detection by PCR and sequencing of the ITS1-5.8S-ITS2 region of the ribosomal DNA (rDNA) revealed infections with F. gigantica (3.38%), Echinostoma caproni (2.36%) and another echinostome (7.09%) that could not be identified further according to its sequence. No dependency of snail size and trematode infection was found. Both high abundance of P. columella in the Fayoum irrigation system and common infection with F. gigantica might be a case of parasite spill-back (increased prevalence in local final hosts due to highly susceptible introduced intermediate host species) from the introduced P. columella to the human population, explaining at least partly the observed increase of reported fascioliasis-cases in Egypt. Eichhornia crassipes, the invasive water hyacinth, which covers huge areas of the irrigation canals, offers safe refuges for the amphibious P. columella during molluscicide application. As a consequence, this snail dominates snail communities and efficiently transmits F. gigantica.

  11. Adjuvant-enhanced antibody and cellular responses to inclusion bodies expressing FhSAP2 correlates with protection of mice to Fasciola hepatica.

    PubMed

    Rivera, Francheska; Espino, Ana M

    2016-01-01

    Fasciola hepatica saposin-like protein-2 (FhSAP2) is a protein differentially expressed in various developmental stages of F. hepatica. Recombinant FhSAP2 has demonstrated the induction of partial protection in mice and rabbits when it is administered subcutaneously (SC) in Freund's adjuvant. Because FhSAP2 is overexpressed in bacteria in the form of inclusion bodies (IBs), we isolated IBs expressing FhSAP2 and tested their immunogenicity when administered SC in mice emulsified in two different adjuvants: QS-21 and Montanide TM ISA720. Animals received three injections containing 20 μg of protein two weeks apart and 4 weeks after the third injection, mice were infected with 10 F. hepatica metacercariae by oral route. The percentages of protection induced by FhSAP2-IBs were estimated to be between 60.0 and 62.5% when compared with adjuvant-vaccinated, infected controls. By determining the levels of IgG1 and IgG2a antibodies and IL-4 and IFNγ cytokines in the serum of experimental animals, it was found that both Th1 and Th2 immune responses were significantly increased in the FhSAP2-IBs vaccinated groups compared with the adjuvant-vaccinated, infected control groups. The adjuvant-vaccinated groups had significantly lower IgG1 to IgG2a ratios and lower IL-4 to IFNγ ratios than the FhSAP2-IBs vaccinated animals, which is indicative of higher levels of Th2 immune responses. Irrespective to the adjuvant used, animals vaccinated with FhSAP2-IBs exhibited significantly higher survival percentage and less liver damage than the adjuvant-control groups. This study suggests that FhSAP2 has potential as vaccine against F. hepatica and that the protection elicited by this molecule could be linked to a mechanism driven by the CD4-Th1 cells. PMID:26632503

  12. Relative importance of management, meteorological and environmental factors in the spatial distribution of Fasciola hepatica in dairy cattle in a temperate climate zone.

    PubMed

    Bennema, S C; Ducheyne, E; Vercruysse, J; Claerebout, E; Hendrickx, G; Charlier, J

    2011-02-01

    Fasciola hepatica, a trematode parasite with a worldwide distribution, is the cause of important production losses in the dairy industry. Diagnosis is hampered by the fact that the infection is mostly subclinical. To increase awareness and develop regionally adapted control methods, knowledge on the spatial distribution of economically important infection levels is needed. Previous studies modelling the spatial distribution of F. hepatica are mostly based on single cross-sectional samplings and have focussed on climatic and environmental factors, often ignoring management factors. This study investigated the associations between management, climatic and environmental factors affecting the spatial distribution of infection with F. hepatica in dairy herds in a temperate climate zone (Flanders, Belgium) over three consecutive years. A bulk-tank milk antibody ELISA was used to measure F. hepatica infection levels in a random sample of 1762 dairy herds in the autumns of 2006, 2007 and 2008. The infection levels were included in a Geographic Information System together with meteorological, environmental and management parameters. Logistic regression models were used to determine associations between possible risk factors and infection levels. The prevalence and spatial distribution of F. hepatica was relatively stable, with small interannual differences in prevalence and location of clusters. The logistic regression model based on both management and climatic/environmental factors included the factors: annual rainfall, mowing of pastures, proportion of grazed grass in the diet and length of grazing season as significant predictors and described the spatial distribution of F. hepatica better than the model based on climatic/environmental factors only (annual rainfall, elevation and slope, soil type), with an Area Under the Curve of the Receiver Operating Characteristic of 0.68 compared with 0.62. The results indicate that in temperate climate zones without large climatic

  13. Fasciola hepatica fatty acid binding protein inhibits TLR4 activation and suppresses the inflammatory cytokines induced by LPS in vitro and in vivo

    PubMed Central

    Martin, Ivelisse; Cabán-Hernández, Kimberly; Figueroa-Santiago, Olgary; Espino, Ana M.

    2015-01-01

    Toll-like receptor 4 (TLR4), the innate immunity receptor for bacterial endotoxins, plays a pivotal role in the induction of inflammatory responses. There is a need to develop molecules that block either activation through TLR4 or the downstream signaling pathways to inhibit the storm of inflammation typically elicited by bacterial lipopolysaccharide (LPS), which is a major cause of the high mortality associated with bacterial sepsis. We report here that a single intraperitoneal injection of 15μg Fasciola hepatica fatty acid binding protein (Fh12) 1 hour before exposure to LPS suppressed significantly the expression of serum inflammatory cytokines in a model of septic shock using C57BL/6 mice. Because macrophages are good source of IL12p70 and TNFα, and critical in driving adaptive immunity, we investigated the effect of Fh12 on the function of mouse bone marrow derived macrophages (bmMΦs). Whereas Fh12 alone did not induce cytokine expression, it significantly suppressed the expression of IL12, TNFα, IL6 and IL1β cytokines as well as iNOS2 in bmMΦs, and also impaired the phagocytic capacity of bmMΦs. Fh12 had a limited effect on the expression of inflammatory cytokines induced in response to other TLR-ligands. One mechanism used by Fh12 to exert its anti-inflammatory effect is binding to the CD14 co-receptor. Moreover, it suppresses phosphorylation of ERK, p38 and JNK. The potent anti-inflammatory properties of Fh12 demonstrated here open doors to further studies directed at exploring the potential of this molecule as a new class of drug against septic shock or other inflammatory diseases. PMID:25780044

  14. Fasciola hepatica Surface Coat Glycoproteins Contain Mannosylated and Phosphorylated N-glycans and Exhibit Immune Modulatory Properties Independent of the Mannose Receptor.

    PubMed

    Ravidà, Alessandra; Aldridge, Allison M; Driessen, Nicole N; Heus, Ferry A H; Hokke, Cornelis H; O'Neill, Sandra M

    2016-04-01

    Fascioliasis, caused by the liver fluke Fasciola hepatica, is a neglected tropical disease infecting over 1 million individuals annually with 17 million people at risk of infection. Like other helminths, F. hepatica employs mechanisms of immune suppression in order to evade its host immune system. In this study the N-glycosylation of F. hepatica's tegumental coat (FhTeg) and its carbohydrate-dependent interactions with bone marrow derived dendritic cells (BMDCs) were investigated. Mass spectrometric analysis demonstrated that FhTeg N-glycans comprised mainly of oligomannose and to a lesser extent truncated and complex type glycans, including a phosphorylated subset. The interaction of FhTeg with the mannose receptor (MR) was investigated. Binding of FhTeg to MR-transfected CHO cells and BMDCs was blocked when pre-incubated with mannan. We further elucidated the role played by MR in the immunomodulatory mechanism of FhTeg and demonstrated that while FhTeg's binding was significantly reduced in BMDCs generated from MR knockout mice, the absence of MR did not alter FhTeg's ability to induce SOCS3 or suppress cytokine secretion from LPS activated BMDCs. A panel of negatively charged monosaccharides (i.e. GlcNAc-4P, Man-6P and GalNAc-4S) were used in an attempt to inhibit the immunoregulatory properties of phosphorylated oligosaccharides. Notably, GalNAc-4S, a known inhibitor of the Cys-domain of MR, efficiently suppressed FhTeg binding to BMDCs and inhibited the expression of suppressor of cytokine signalling (SOCS) 3, a negative regulator the TLR and STAT3 pathway. We conclude that F. hepatica contains high levels of mannose residues and phosphorylated glycoproteins that are crucial in modulating its host's immune system, however the role played by MR appears to be limited to the initial binding event suggesting that other C-type lectin receptors are involved in the immunomodulatory mechanism of FhTeg.

  15. The feasibility of testing whether Fasciola hepatica is associated with increased risk of verocytotoxin producing Escherichia coli O157 from an existing study protocol.

    PubMed

    Hickey, Graeme L; Diggle, Peter J; McNeilly, Tom N; Tongue, Sue C; Chase-Topping, Margo E; Williams, Diana J L

    2015-05-01

    The parasite Fasciola hepatica is a major cause of economic loss to the agricultural community worldwide as a result of morbidity and mortality in livestock, including cattle. Cattle are the principle reservoir of verocytotoxigenic Escherichia coli O157 (VTEC O157), an important cause of disease in humans. To date there has been little empirical research on the interaction between F. hepatica and VTEC O157. It is hypothesised that F. hepatica, which is known to suppress type 1 immune responses and induce an anti-inflammatory or regulatory immune environment in the host, may promote colonisation of the bovine intestine with VTEC O157. Here we assess whether it is statistically feasible to augment a prospective study to quantify the prevalence of VTEC O157 in cattle in Great Britain with a pilot study to test this hypothesis. We simulate data under the framework of a mixed-effects logistic regression model in order to calculate the power to detect an association effect size (odds ratio) of 2. In order to reduce the resources required for such a study, we exploit the fact that the test results for VTEC O157 will be known in advance of testing for F. hepatica by restricting analysis to farms with a VTEC O157 sample prevalence of >0% and <100%. From a total of 270 farms (mean 27 cows per farm) that will be tested for VTEC O157, power of 87% can be achieved, whereby testing of F. hepatica would only be necessary for an expected 50 farms, thus considerably reducing costs. Pre-study sample size calculations are an important part of any study design. The framework developed here is applicable to the study of other co-infections.

  16. Adjuvant-enhanced antibody and cellular responses to inclusion bodies expressing FhSAP2 correlates with protection of mice to Fasciola hepatica.

    PubMed

    Rivera, Francheska; Espino, Ana M

    2016-01-01

    Fasciola hepatica saposin-like protein-2 (FhSAP2) is a protein differentially expressed in various developmental stages of F. hepatica. Recombinant FhSAP2 has demonstrated the induction of partial protection in mice and rabbits when it is administered subcutaneously (SC) in Freund's adjuvant. Because FhSAP2 is overexpressed in bacteria in the form of inclusion bodies (IBs), we isolated IBs expressing FhSAP2 and tested their immunogenicity when administered SC in mice emulsified in two different adjuvants: QS-21 and Montanide TM ISA720. Animals received three injections containing 20 μg of protein two weeks apart and 4 weeks after the third injection, mice were infected with 10 F. hepatica metacercariae by oral route. The percentages of protection induced by FhSAP2-IBs were estimated to be between 60.0 and 62.5% when compared with adjuvant-vaccinated, infected controls. By determining the levels of IgG1 and IgG2a antibodies and IL-4 and IFNγ cytokines in the serum of experimental animals, it was found that both Th1 and Th2 immune responses were significantly increased in the FhSAP2-IBs vaccinated groups compared with the adjuvant-vaccinated, infected control groups. The adjuvant-vaccinated groups had significantly lower IgG1 to IgG2a ratios and lower IL-4 to IFNγ ratios than the FhSAP2-IBs vaccinated animals, which is indicative of higher levels of Th2 immune responses. Irrespective to the adjuvant used, animals vaccinated with FhSAP2-IBs exhibited significantly higher survival percentage and less liver damage than the adjuvant-control groups. This study suggests that FhSAP2 has potential as vaccine against F. hepatica and that the protection elicited by this molecule could be linked to a mechanism driven by the CD4-Th1 cells.

  17. Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach.

    PubMed

    Cwiklinski, Krystyna; Allen, Katherine; LaCourse, James; Williams, Diana J; Paterson, Steve; Hodgkinson, Jane E

    2015-06-01

    The liver fluke, Fasciola hepatica is an economically important pathogen of sheep and cattle and has been described by the WHO as a re-emerging zoonosis. Control is heavily reliant on the use of drugs, particularly triclabendazole and as a result resistance has now emerged. The population structure of F. hepatica is not well known, yet it can impact on host-parasite interactions and parasite control with drugs, particularly regarding the spread of triclabendazole resistance. We have identified 2448 potential microsatellites from 83 Mb of F. hepatica genome sequence using msatfinder. Thirty-five loci were developed and optimised for microsatellite PCR, resulting in a panel of 15 polymorphic loci, with a range of three to 15 alleles. This panel was validated on genomic DNA from 46 adult F. hepatica; 38 liver flukes sourced from a Northwest abattoir, UK and 8 liver flukes from an established isolate (Shrewsbury; Ridgeway Research). Evidence for null alleles was found at four loci (Fh_1, Fh_8, Fh_13 and Fh_14), which showed markedly higher levels of homozygosity than the remaining 11 loci. Of the 38 liver flukes isolated from cattle livers (n=10) at the abattoir, 37 genotypes were identified. Using a multiplex approach all 15 loci could be amplified from several life cycle stages that typically yield low amounts of DNA, including metacercariae, the infective life cycle stage present on pasture, highlighting the utility of this multiplex microsatellite panel. This study reports the largest panel of microsatellite markers available to date for population studies of F. hepatica and the first multiplex panel of microsatellite markers that can be used for several life cycle stages.

  18. The feasibility of testing whether Fasciola hepatica is associated with increased risk of verocytotoxin producing Escherichia coli O157 from an existing study protocol

    PubMed Central

    Hickey, Graeme L.; Diggle, Peter J.; McNeilly, Tom N.; Tongue, Sue C.; Chase-Topping, Margo E.; Williams, Diana J.L.

    2015-01-01

    The parasite Fasciola hepatica is a major cause of economic loss to the agricultural community worldwide as a result of morbidity and mortality in livestock, including cattle. Cattle are the principle reservoir of verocytotoxigenic Escherichia coli O157 (VTEC O157), an important cause of disease in humans. To date there has been little empirical research on the interaction between F. hepatica and VTEC O157. It is hypothesised that F. hepatica, which is known to suppress type 1 immune responses and induce an anti-inflammatory or regulatory immune environment in the host, may promote colonisation of the bovine intestine with VTEC O157. Here we assess whether it is statistically feasible to augment a prospective study to quantify the prevalence of VTEC O157 in cattle in Great Britain with a pilot study to test this hypothesis. We simulate data under the framework of a mixed-effects logistic regression model in order to calculate the power to detect an association effect size (odds ratio) of 2. In order to reduce the resources required for such a study, we exploit the fact that the test results for VTEC O157 will be known in advance of testing for F. hepatica by restricting analysis to farms with a VTEC O157 sample prevalence of >0% and <100%. From a total of 270 farms (mean 27 cows per farm) that will be tested for VTEC O157, power of 87% can be achieved, whereby testing of F. hepatica would only be necessary for an expected 50 farms, thus considerably reducing costs. Pre-study sample size calculations are an important part of any study design. The framework developed here is applicable to the study of other co-infections. PMID:25779556

  19. The use of bulk-tank milk ELISAs to assess the spatial distribution of Fasciola hepatica, Ostertagia ostertagi and Dictyocaulus viviparus in dairy cattle in Flanders (Belgium).

    PubMed

    Bennema, S; Vercruysse, J; Claerebout, E; Schnieder, T; Strube, C; Ducheyne, E; Hendrickx, G; Charlier, J

    2009-10-28

    Fasciola hepatica, Ostertagia ostertagi and Dictyocaulus viviparus are helminth parasites with a wide distribution and an important economic impact in cattle in temperate climates. This paper describes the spatial distribution of F. hepatica, O. ostertagi and D. viviparus in dairy herds in Flanders (Belgium). One thousand eight hundred herds were selected at random from the Flemish dairy population (n=7002), stratified on community level to obtain a sample representative for the entire study area. From each herd, a bulk milk sample collected in autumn 2006 was analysed with previously described antibody-ELISAs in order to identify herds where the parasite infection level is likely to cause production loss (F. hepatica and O. ostertagi) (defined as economic infections) or where patent infections have been present over the past grazing season (D. viviparus). The herd prevalence of economic infections with F. hepatica and O. ostertagi was 37.3% (95% Confidence Interval (CI): 35.1-39.7) and 59.1% (95%CI: 56.8-61.4), respectively. The herd prevalence of D. viviparus was 19.6% (95%CI: 17.7-21.6). On 28.9% (CI 26.8-31.3) of the herds, low levels of infection were observed for all three of the helminths. The presence of clustering of (economic) infections was studied using Moran's I, whereas the location and size of the clusters were studied using the spatial scan statistic, the Local Indicator of Spatial Association and Kernel density plotting. A marked clustering in the spatial distribution of F. hepatica and a mild clustering in the spatial distribution of O. ostertagi were observed. D. viviparus infections were spread evenly over Flanders. Knowledge of locations of high risk areas can lead to increased awareness and may be the start of the development of regionally adapted control measures.

  20. The miRnome of Fasciola hepatica juveniles endorses the existence of a reduced set of highly divergent micro RNAs in parasitic flatworms.

    PubMed

    Fontenla, Santiago; Dell'Oca, Nicolás; Smircich, Pablo; Tort, José F; Siles-Lucas, Mar

    2015-12-01

    The liver fluke Fasciola hepatica is a foodborne zoonotic parasite affecting livestock worldwide, with increasing relevance in human health. The first developmental stage that the host meets after ingestion of the parasite is the newly excysted juvenile, that actively transverses the gut wall and migrates to its final location in the liver. The regulation of the early developmental events in newly excysted juveniles is still poorly understood and a relevant target for control strategies. Here we investigated the putative involvement of small regulatory RNAs in the invasion process. The small RNA population of the newly excysted juvenile fall into two classes, one represented by micro (mi)RNAs and a secondary group of larger (32-33 nucleotides) tRNA-derived sequences. We identified 40 different miRNAs, most of those belonging to ancient miRNAs conserved in protostomes and metazoans, notably with a highly predominant miR-125b variant. Remarkably, several protostomian and metazoan conserved families were not detected in consonance with previous reports of drastic miRnome reduction in parasitic flatworms. Additionally, a set of five novel miRNAs was identified, probably associated with specific gene regulation expression needs in F. hepatica. While sequence conservation in mature miRNA is high across the metazoan tree, we observed that flatworm miRNAs are more divergent, suggesting that mutation rates in parasitic flatworms could be high. Finally, the distinctive presence of tRNA-derived sequences, mostly 5' tRNA halves of selected tRNAs in the small RNA population of newly excysted juveniles, raises the possibility that both miRNA and tRNA fragments participate in the regulation of gene expression in this parasite.

  1. Effect of induced Fasciola gigantica infection during pre-patency on the performance of buffalo calves fed on different percentage of protein.

    PubMed

    Singh, P; Verma, A K; Gupta, S C; Mehra, U R

    2016-09-01

    Thirty growing Murrah buffalo calves (8-12 months of age, 109.85 ± 2.43) were reared in parasite free conditions and randomly divided into three equal groups as per CRD. They were fed on iso-caloric (2.01 ME Mcal/Kg diet) diets containing standard protein (SP) diet at 100 %, 90 % of SP (medium protein, MP) and 80 % of SP (low protein, LP) of the protein requirements (Kearl 1982). After 21 days of feeding, each group was further subdivided into two sub-groups (A & B). Animals in sub-groups 'A' served as non-infected control, while in sub group 'B' were orally infected with Fasciola gigantica metacercarie (mc; 1,000 each). A metabolic trial of 40 days post infection was carried out in control and parasitized animals. Intake of digestible dry matter, organic matter and acid detergent fibre (ADF) was significantly higher (P < 0.05) in SP group compared to LP group. The digestibility of crude protein (CP) and ADF was significantly higher in SP group compared to MP and LP groups. The digestible crude protein (DCP) and total digestible nutrients (TDN) intakes (g/kgW(0.75)) were also significantly (P < 0.001) higher in SP than MP and LP groups. However, DCP intake was significantly (P < 0.001) lower in infected subgroups compared to control subgroups. Intake and balance (g/d) of nitrogen, calcium and phosphorus were significantly (P < 0.01) higher in SP than MP and LP groups. The average daily gain of buffalo calves fed on SP Uninfected (SPU), SP Infected (SPI), MPU, MPI and LPU, LPI groups was 333, 178, 356, 144, 222 and 144 g and was significantly (P < 0.01) lower in animals fed LP ration. The feed conversion ratio (FCR) was also significantly (P < 0.01) higher in infected sub-groups as compared to respective control groups. The results showed that a SP diet substantially improved the overall performance of buffalo calves in control and infected groups and reduced adverse effect of F. gigantica infection. PMID:27605773

  2. Fasciola hepatica Surface Coat Glycoproteins Contain Mannosylated and Phosphorylated N-glycans and Exhibit Immune Modulatory Properties Independent of the Mannose Receptor

    PubMed Central

    Ravidà, Alessandra; Aldridge, Allison M.; Driessen, Nicole N.; Heus, Ferry A. H.; Hokke, Cornelis H.; O’Neill, Sandra M.

    2016-01-01

    Fascioliasis, caused by the liver fluke Fasciola hepatica, is a neglected tropical disease infecting over 1 million individuals annually with 17 million people at risk of infection. Like other helminths, F. hepatica employs mechanisms of immune suppression in order to evade its host immune system. In this study the N-glycosylation of F. hepatica’s tegumental coat (FhTeg) and its carbohydrate-dependent interactions with bone marrow derived dendritic cells (BMDCs) were investigated. Mass spectrometric analysis demonstrated that FhTeg N-glycans comprised mainly of oligomannose and to a lesser extent truncated and complex type glycans, including a phosphorylated subset. The interaction of FhTeg with the mannose receptor (MR) was investigated. Binding of FhTeg to MR-transfected CHO cells and BMDCs was blocked when pre-incubated with mannan. We further elucidated the role played by MR in the immunomodulatory mechanism of FhTeg and demonstrated that while FhTeg’s binding was significantly reduced in BMDCs generated from MR knockout mice, the absence of MR did not alter FhTeg’s ability to induce SOCS3 or suppress cytokine secretion from LPS activated BMDCs. A panel of negatively charged monosaccharides (i.e. GlcNAc-4P, Man-6P and GalNAc-4S) were used in an attempt to inhibit the immunoregulatory properties of phosphorylated oligosaccharides. Notably, GalNAc-4S, a known inhibitor of the Cys-domain of MR, efficiently suppressed FhTeg binding to BMDCs and inhibited the expression of suppressor of cytokine signalling (SOCS) 3, a negative regulator the TLR and STAT3 pathway. We conclude that F. hepatica contains high levels of mannose residues and phosphorylated glycoproteins that are crucial in modulating its host’s immune system, however the role played by MR appears to be limited to the initial binding event suggesting that other C-type lectin receptors are involved in the immunomodulatory mechanism of FhTeg. PMID:27104959

  3. Insights into the Interactions of Fasciola hepatica Cathepsin L3 with a Substrate and Potential Novel Inhibitors through In Silico Approaches

    PubMed Central

    Hernández Alvarez, Lilian; Naranjo Feliciano, Dany; Hernández González, Jorge Enrique; de Oliveira Soares, Rosemberg; Barreto Gomes, Diego Enry; Pascutti, Pedro Geraldo

    2015-01-01

    Background Fasciola hepatica is the causative agent of fascioliasis, a disease affecting grazing animals, causing economic losses in global agriculture and currently being an important human zoonosis. Overuse of chemotherapeutics against fascioliasis has increased the populations of drug resistant parasites. F. hepatica cathepsin L3 is a protease that plays important roles during the life cycle of fluke. Due to its particular collagenolytic activity it is considered an attractive target against the infective phase of F. hepatica. Methodology/Principal Findings Starting with a three dimensional model of FhCL3 we performed a structure-based design of novel inhibitors through a computational study that combined virtual screening, molecular dynamics simulations, and binding free energy (ΔGbind) calculations. Virtual screening was carried out by docking inhibitors obtained from the MYBRIDGE-HitFinder database inside FhCL3 and human cathepsin L substrate-binding sites. On the basis of dock-scores, five compounds were predicted as selective inhibitors of FhCL3. Molecular dynamic simulations were performed and, subsequently, an end-point method was employed to predict ΔGbind values. Two compounds with the best ΔGbind values (-10.68 kcal/mol and -7.16 kcal/mol), comparable to that of the positive control (-10.55 kcal/mol), were identified. A similar approach was followed to structurally and energetically characterize the interface of FhCL3 in complex with a peptidic substrate. Finally, through pair-wise and per-residue free energy decomposition we identified residues that are critical for the substrate/ligand binding and for the enzyme specificity. Conclusions/Significance The present study is the first computer-aided drug design approach against F. hepatica cathepsins. Here we predict the principal determinants of binding of FhCL3 in complex with a natural substrate by detailed energetic characterization of protease interaction surface. We also propose novel compounds

  4. Prevalence of Liver Fluke (Fasciola hepatica) in Wild Red Deer (Cervus elaphus): Coproantigen ELISA Is a Practicable Alternative to Faecal Egg Counting for Surveillance in Remote Populations.

    PubMed

    French, Andrew S; Zadoks, Ruth N; Skuce, Philip J; Mitchell, Gillian; Gordon-Gibbs, Danielle K; Craine, Alexandra; Shaw, David; Gibb, Stuart W; Taggart, Mark A

    2016-01-01

    Red deer (Cervus elaphus) are hosts of liver fluke (Fasciola hepatica); yet, prevalence is rarely quantified in wild populations. Testing fresh samples from remote regions by faecal examination (FE) can be logistically challenging; hence, we appraise frozen storage and the use of a coproantigen ELISA (cELISA) for F. hepatica surveillance. We also present cELISA surveillance data for red deer from the Highlands of Scotland. Diagnoses in faecal samples (207 frozen, 146 fresh) were compared using a cELISA and by FE. For each storage method (frozen or fresh), agreement between the two diagnostics was estimated at individual and population levels, where population prevalence was stratified into cohorts (e.g., by sampling location). To approximate sensitivity and specificity, 65 post-slaughter whole liver examinations were used as a reference. At the individual level, FE and cELISA diagnoses agreed moderately (κfrozen = 0.46; κfresh = 0.51), a likely reflection of their underlying principles. At the population level, FE and cELISA cohort prevalence correlated strongly (Pearson's R = 0.89, p < 0.0001), reflecting good agreement on relative differences between cohort prevalence. In frozen samples, prevalence by cELISA exceeded FE overall (42.8% vs. 25.8%) and in 9/12 cohorts, alluding to differences in sensitivity; though, in fresh samples, no significant difference was found. In 959 deer tested by cELISA across the Scottish Highlands, infection prevalence ranged from 9.6% to 53% by sampling location. We highlight two key advantages of cELISA over FE: i) the ability to store samples long term (frozen) without apparent loss in diagnostic power; and ii) reduced labour and the ability to process large batches. Further evaluation of cELISA sensitivity in red deer, where a range of fluke burdens can be obtained, is desirable. In the interim, the cELISA is a practicable diagnostic for F. hepatica surveillance in red deer, and its application here has revealed considerable

  5. A comparative study on the impact of two artemisinin derivatives, artemether and artesunate, on the female reproductive system of Fasciola hepatica.

    PubMed

    O'Neill, J F; Johnston, R C; Halferty, L; Hanna, R E B; Brennan, G P; Fairweather, I

    2015-07-30

    An in vivo study in the laboratory rat model has been carried out to monitor changes to the female reproductive system in adult Fasciola hepatica following treatment with the artemisinins, artemether and artesunate. Rats infected with the triclabendazole (TCBZ)-resistant Sligo isolate were dosed orally with artemether at a concentration of 200mg/kg and flukes recovered at 24, 48 and 72 h post-treatment (pt). Rats infected with the TCBZ-resistant Oberon isolate were dosed orally with artesunate at a concentration of 200mg/kg and flukes recovered 24, 48, 72 and 96 h pt. The flukes were processed for histological and transmission electron microscope (TEM) examination of the uterus, Mehlis' gland, ovary and vitellaria. After treatment with artemether, egg production had become abnormal by 72 h pt, with free vitelline cells and masses of shell protein material within the uterus; spermatozoa were absent. The Mehlis' gland and ovary retained a normal morphology over the 3-day period. A change in the cell population in the vitelline follicles was seen at 48 h pt, with a decline in the number of immature cells. This became more marked by 72 h and the follicles became progressively vacuolated over the 3-day period. At the TEM level, there were changes in the immature vitelline cells at 24h pt, as evidenced by a decrease in shell protein production and the presence of lipid droplets and abnormal mitochondria. Spaces in the follicles separated the cells from each other. The changes became progressively more severe with time, so that, by 72 h pt, the follicles were very disrupted, containing cells in the advanced stages of apoptotic breakdown. In extreme cases, the follicles were scarcely recognisable and had become filled with cellular debris. Fine structural changes to the vitelline cells induced by artesunate treatment were similar to those described for artemether, but generally occurred more quickly and were greater; this was particularly true of the swelling of the ger

  6. Insights into the biological features of the antigenic determinants recognized by four monoclonal antibodies in redia and adult stages of the liver fluke Fasciola hepatica.

    PubMed

    Alba, Annia; Sánchez, Jorge; Hernández, Hilda; Mosqueda, Maryani; Rodríguez, Suanel Y; Capó, Virginia; Otero, Oscar; Alfonso, Carlos; Marcet, Ricardo; Sarracent, Jorge

    2016-09-01

    Fasciola hepatica is a digenean trematode which infects a wide variety of domestic animals and also humans. Previous studies have demonstrated that four monoclonal antibodies (Mabs) against the total extract of F. hepatica redia (named as 1E4, 6G11, 4E5 and 4G11) also recognized the excretion - secretion antigens (ES Ag) of adult parasites, which is a biologically-relevant mixture of molecules with functional roles during infection and immune evasion on definitive hosts. In the present report we describe the partial characterization of the epitopes recognized by these Mabs by heat treatment, mercaptoethanol reduction, pronase proteolysis and sodium peryodate oxidation, which suggested their predominant protein and conformational nature. Also, a comparative study using immunodetection assays on crude extracts and on histological sections of both rediae and adults of F. hepatica were performed to explore the expression pattern of the antigenic determinants in these developmental stages. From these experiments it was found that the Mabs reacted most likely with the same proteins of approximately 64 and 105 kDa present on both rediae and adult's extracts. However, the 1E4, 6G11 and 4E5 Mabs also recognized other molecules of the total extract of F. hepatica adults, a fact that constitutes an evidence of the antigenic variation between both stages and points at a certain biological relevance of the recognized antigenic determinants. Immunolocalization studies on histological sections revealed that all Mabs reacted with the tegument of F. hepatica in both rediae and adults stages, while the epitopes recognized by 1E4, 6G11 and 4E5 antibodies were also preferentially localized in the intestinal caeca and in different organs of the reproductive system of adult specimens. The immunogenicity of these antigenic determinants, their conserved status among different stages of the life cycle of F. hepatica and their presence in both tegument and ES Ag of adult parasites

  7. High prevalence of fasciolosis and evaluation of drug efficacy against Fasciola hepatica in dairy cattle in the Maffra and Bairnsdale districts of Gippsland, Victoria, Australia.

    PubMed

    Elliott, T P; Kelley, J M; Rawlin, G; Spithill, T W

    2015-04-15

    Liver fluke (Fasciola hepatica) is a common parasite amongst grazing livestock in the south-eastern region of Australia and is responsible for significant production losses in the beef and dairy industries. Gippsland in Victoria is a major region for dairy production but no fluke prevalence data in livestock has been obtained in this region since the late 1970s prior to the introduction of Triclabendazole (TCBZ). TCBZ resistance is also now widespread in cattle in south east Australia. In this study, we evaluated the prevalence and intensity of liver fluke infections in dairy cattle in Gippsland and assessed the efficacy of TCBZ and other drenches against F. hepatica on one farm. We obtained 30 individual faecal samples from each of 15 different farms and, using the liver fluke coproantigen ELISA, tested bulk faecal samples pooled from each farm. Any farm that returned a positive bulk sample had all of the samples tested individually to assess the intra-herd prevalence. One farm in the Maffra district also had a coproantigen reduction test and faecal egg count reduction test to assess the efficacy of TCBZ, Clorsulon (CLOR) and Oxyclozanide (OXY). The coproantigen ELISA proved to be a highly sensitive test for liver fluke with a high correlation (R(2)=0.8849) observed between ELISA data from bulk samples and individual samples, suggesting that future larger scale screening on farms for fasciolosis could use the bulk analysis technique. The ELISA data revealed that animals on six of the 15 farms were infected with F. hepatica and the herd prevalence of the infected herds ranged from 47 to 100% (mean 81%) which exceeds the prevalence value for production losses of 25%. The intensity of fluke infection in cattle varied considerably both within and between herds with a proportion of animals exhibiting a positive control value in the coproantigen ELISA of 50-88%. We also confirmed that TCBZ resistance was present on one farm but that CLOR or OXY can be used to remove the

  8. The contamination of wild watercress with Fasciola hepatica in central France depends on the ability of several lymnaeid snails to migrate upstream towards the beds.

    PubMed

    Rondelaud, D; Hourdin, P; Vignoles, P; Dreyfuss, G

    2005-03-01

    As most natural watercress beds in central France are located upstream of the permanent habitats of two lymnaeid species, Galba truncatula and Omphiscola glabra, field investigations were made from 1999 to 2004 on 67 beds to determine why the contamination of watercress with Fasciola hepatica is irregular over time in these sites, while definitive hosts, especially lagomorphs, are regularly found infected around them. Snails are able to migrate upstream in winter and spring towards the beds, and a 4-year survey demonstrated the existence of annual variation in the colonization of these sites by snails. In the 45 beds irregularly contaminated with F. hepatica over time, 37.7-62.2%, according to the year, were not populated by lymnaeids, in spite of the presence of snail populations downstream. G. truncatula was found alone in 8.8-13.3% of sites and was the first colonizing snail in 24.3-33.3% when the two lymnaeid species successively settled in these waterholes. The colonizing ability of O. glabra was more limited, as it was observed alone in 2.2% of beds and was the first colonizing snail in only 2.2-20% of them. The distances covered by these snails significantly increased with increasing migration time. After migration, a few overwintering snails (3.8% of G. truncatula and 6.8% of O. glabra) are able to colonize the beds, but their numbers decreased when the distance of migration was greater. O. glabra migrated more quickly and reached more watercress beds than G. truncatula. However, this did not influence the natural infections of snails, which were more frequent in the F1 of G. truncatula. The variability observed in the colonization of beds by snails might explain the regular or irregular contamination of wild watercress by metacercariae of F. hepatica. One of the factors which may explain this variability is the occurrence of showers in spring, so that a few snails are able to reach watercress beds after their upstream migration.

  9. Prevalence of Liver Fluke (Fasciola hepatica) in Wild Red Deer (Cervus elaphus): Coproantigen ELISA Is a Practicable Alternative to Faecal Egg Counting for Surveillance in Remote Populations

    PubMed Central

    French, Andrew S.; Zadoks, Ruth N.; Skuce, Philip J.; Mitchell, Gillian; Gordon-Gibbs, Danielle K.; Craine, Alexandra; Shaw, David; Gibb, Stuart W.; Taggart, Mark A.

    2016-01-01

    Red deer (Cervus elaphus) are hosts of liver fluke (Fasciola hepatica); yet, prevalence is rarely quantified in wild populations. Testing fresh samples from remote regions by faecal examination (FE) can be logistically challenging; hence, we appraise frozen storage and the use of a coproantigen ELISA (cELISA) for F. hepatica surveillance. We also present cELISA surveillance data for red deer from the Highlands of Scotland. Diagnoses in faecal samples (207 frozen, 146 fresh) were compared using a cELISA and by FE. For each storage method (frozen or fresh), agreement between the two diagnostics was estimated at individual and population levels, where population prevalence was stratified into cohorts (e.g., by sampling location). To approximate sensitivity and specificity, 65 post-slaughter whole liver examinations were used as a reference. At the individual level, FE and cELISA diagnoses agreed moderately (κfrozen = 0.46; κfresh = 0.51), a likely reflection of their underlying principles. At the population level, FE and cELISA cohort prevalence correlated strongly (Pearson’s R = 0.89, p < 0.0001), reflecting good agreement on relative differences between cohort prevalence. In frozen samples, prevalence by cELISA exceeded FE overall (42.8% vs. 25.8%) and in 9/12 cohorts, alluding to differences in sensitivity; though, in fresh samples, no significant difference was found. In 959 deer tested by cELISA across the Scottish Highlands, infection prevalence ranged from 9.6% to 53% by sampling location. We highlight two key advantages of cELISA over FE: i) the ability to store samples long term (frozen) without apparent loss in diagnostic power; and ii) reduced labour and the ability to process large batches. Further evaluation of cELISA sensitivity in red deer, where a range of fluke burdens can be obtained, is desirable. In the interim, the cELISA is a practicable diagnostic for F. hepatica surveillance in red deer, and its application here has revealed considerable

  10. Characterization of the humoral immune response in alpacas (Lama pacos) experimentally infected with Fasciola hepatica against cysteine proteinases Fas1 and Fas2 and histopathological findings.

    PubMed

    Timoteo, O; Maco, V; Maco, V; Neyra, V; Yi, P J; Leguía, G; Espinoza, J R

    2005-06-15

    A characterization of the humoral immune response of alpacas to Fasciola hepatica Fas1 and Fas2 antigens, two abundant cysteine proteinases in the excretory/secretory (E/S) products, was performed over the course of 6 months of experimental infection. Six adult alpacas aged 1-2 years old received a single dose of 200 F. hepatica metacercariae; two non-infected alpacas were kept as control group. All infected animals shed eggs 8 weeks post-infection (PI) and the number of flukes recovered at necropsy averaged 41+/-4. The livers of infected animals showed regions with chronic inflammation, granuloma containing parasite eggs, necrosis and cirrhosis. Peripheral eosinophilia in infected animals was greatly enhanced 6 weeks post-infection and later. A single peak of serum glutamic piruvic transaminase (SGPT) was observed 4 weeks PI and serum glutamic oxalacetic transaminase (SGOT) elevated 3 weeks PI and later. Circulating IgG Abs against Fas1 and Fas2 were measured by enzyme-linked immunosorbent assay (ELISA). Fas2-ELISA detected the infection 10 days PI reaching to highest titer on 7-8 weeks PI and kept elevated, until the end of infection. Fas1-ELISA detected the infection 2 weeks PI and followed the same pattern as Fas2-ELISA. Anti Fas2 IgG Abs were in higher titers and showed stronger avidity than anti Fas1 IgG Abs. In addition, rabbit IgG antibodies raised against cysteine proteinase Fas2 showed infiltration of this parasite antigen associated to the degradation of bile ducts and liver parenchyma of infected alpacas. In the present study we have established a F. hepatica experimental infection of alpacas, Fas2 appears to have a role in the pathogenesis of the liver damage in alpacas caused by the liver fluke. Infected alpacas elicited a strong humoral immune response against fluke cysteine proteinases Fas1 and Fas2, which might be considered as candidates for immunodiagnosis and vaccine development against fasciolosis in alpacas.

  11. Fasciola hepatica excretory-secretory products induce CD4+T cell anergy via selective up-regulation of PD-L2 expression on macrophages in a Dectin-1 dependent way.

    PubMed

    Guasconi, Lorena; Chiapello, Laura S; Masih, Diana T

    2015-07-01

    Fasciola hepatica excretory-secretory products (FhESP) induce immunomodulatory effects on macrophages. Previously, we demonstrated that these effects are dependent on Dectin-1. Therefore, the aim of this study was to determine how this affects the CD4 T-cells immune response. We observed that FhESP induce an increased expression of PD-L2 in macrophages via Dectin-1. Furthermore, in co-cultures with CD4 T-cell we observed a suppressive effect on proliferative response, down-modulation of IFN-γ and up-modulation of IL-10 via Dectin-1 on macrophages. These results suggest that FhESP induce T-cell anergy via selective up-regulation of PD-L2 expression on macrophages in a Dectin-1 dependent way.

  12. In Vitro and In Vivo Studies for Assessing the Immune Response and Protection-Inducing Ability Conferred by Fasciola hepatica-Derived Synthetic Peptides Containing B- and T-Cell Epitopes

    PubMed Central

    Rojas-Caraballo, Jose; López-Abán, Julio; Pérez del Villar, Luis; Vizcaíno, Carolina; Vicente, Belén; Fernández-Soto, Pedro; del Olmo, Esther; Patarroyo, Manuel Alfonso; Muro, Antonio

    2014-01-01

    Fasciolosis is considered the most widespread trematode disease affecting grazing animals around the world; it is currently recognised by the World Health Organisation as an emergent human pathogen. Triclabendazole is still the most effective drug against this disease; however, resistant strains have appeared and developing an effective vaccine against this disease has increasingly become a priority. Several bioinformatics tools were here used for predicting B- and T-cell epitopes according to the available data for Fasciola hepatica protein amino acid sequences. BALB/c mice were immunised with the synthetic peptides by using the ADAD vaccination system and several immune response parameters were measured (antibody titres, cytokine levels, T-cell populations) to evaluate their ability to elicit an immune response. Based on the immunogenicity results so obtained, seven peptides were selected to assess their protection-inducing ability against experimental infection with F. hepatica metacercariae. Twenty-four B- or T-epitope-containing peptides were predicted and chemically synthesised. Immunisation of mice with peptides so-called B1, B2, B5, B6, T14, T15 and T16 induced high levels of total IgG, IgG1 and IgG2a (p<0.05) and a mixed Th1/Th2/Th17/Treg immune response, according to IFN-γ, IL-4, IL-17 and IL-10 levels, accompanied by increased CD62L+ T-cell populations. A high level of protection was obtained in mice vaccinated with peptides B2, B5, B6 and T15 formulated in the ADAD vaccination system with the AA0029 immunomodulator. The bioinformatics approach used in the present study led to the identification of seven peptides as vaccine candidates against the infection caused by Fasciola hepatica (a liver-fluke trematode). However, vaccine efficacy must be evaluated in other host species, including those having veterinary importance. PMID:25122166

  13. Evidence for high genetic diversity of NAD1 and COX1 mitochondrial haplotypes among triclabendazole resistant and susceptible populations and field isolates of Fasciola hepatica (liver fluke) in Australia.

    PubMed

    Elliott, T; Muller, A; Brockwell, Y; Murphy, N; Grillo, V; Toet, H M; Anderson, G; Sangster, N; Spithill, T W

    2014-02-24

    In recent years, the global incidence of Fasciola hepatica (liver fluke) infections exhibiting resistance to triclabendazole (TCBZ) has increased, resulting in increased economic losses for livestock producers and threatening future control. The development of TCBZ resistance and the worldwide discovery of F. hepatica population diversity has emphasized the need to further understand the genetic structure of drug susceptible and resistant Fasciola populations within Australia. In this study, the genetic diversity of liver flukes was estimated by sequencing mitochondrial DNA (mtDNA) encoding the NAD1 (530 bp) and COX1 (420 bp) genes of 208 liver flukes (F. hepatica) collected from three populations: field isolates obtained from abattoirs from New South Wales (NSW) and Victoria (Vic); three TCBZ-resistant fluke populations from NSW and Victoria; and the well-established TCBZ-susceptible Sunny Corner laboratory isolate. Overall nucleotide diversity for all flukes analysed of 0.00516 and 0.00336 was estimated for the NAD1 and COX1 genes respectively. Eighteen distinct haplotypes were established for the NAD1 gene and six haplotypes for the COX1 gene, resulting in haplotype diversity levels of 0.832 and 0.482, respectively. One field isolate showed a similar low level of haplotype diversity as seen in the Sunny Corner laboratory isolate. Analysis of TCBZ-resistant infrapopulations from 3 individual cattle grazing one property revealed considerable sequence parasite diversity between cattle. Analysis of parasite TCBZ-resistant infrapopulations from sheep and cattle revealed haplotypes unique to each host, but no significant difference between parasite populations. Fst analysis of fluke populations revealed little differentiation between the resistant and field populations. This study has revealed a high level of diversity in field and drug resistant flukes in South-Eastern Australia.

  14. Fasciola hepatica: Histology of the testis in egg-producing adults of several laboratory-maintained isolates of flukes grown to maturity in cattle and sheep and in flukes from naturally infected hosts.

    PubMed

    Hanna, R E B; Edgar, H; Moffett, D; McConnell, S; Fairweather, I; Brennan, G P; Trudgett, A; Hoey, E M; Cromie, L; Taylor, S M; Daniel, R

    2008-11-01

    A total of 8 calves approximately 6 months old and 22 lambs of similar age were infected with metacercariae of Fasciola hepatica of various laboratory-maintained isolates including: Cullompton (sensitive to triclabendazole) and Sligo, Oberon and Leon (reported as resistant to triclabendazole). Ten to 16 weeks after infection, flukes were harvested from these experimental animals and the histology of the testis tissue was examined in a representative sample of flukes from each population. Adult wild-type flukes were also collected from 5 chronically infected cattle and 7 chronically infected sheep identified at post-mortem inspection. The testis tissue of these flukes was compared with that of the various laboratory-maintained isolates. Whilst the testes of the wild-type, Oberon and Leon flukes displayed all the usual cell types associated with spermatogenesis in Fasciola hepatica (spermatogonia, spermatocytes, spermatids and mature sperm), the Cullompton flukes from both cattle and sheep showed arrested spermatogenesis, with no stages later than primary spermatocytes represented in the testis profiles. The presence of numerous eosinophilic apoptotic bodies and nuclear fragments suggested that meiotic division was anomalous and incomplete. In contrast to the wild-type flukes, no mature spermatozoa were present in the testes or amongst the shelled eggs in the uterus. A high proportion of the eggs collected from these flukes hatched to release normal-appearing miracidia after an appropriate incubation period, as indeed was the case with all isolates examined and the wild-type flukes. It is concluded that the eggs of Cullompton flukes are capable of development without fertilization, i.e. are parthenogenetic. The implications of this for rapid evolution of resistant clones following an anthelmintic selection event are discussed. Amongst the Sligo flukes examined, two subtypes were recognised, namely, those flukes with all stages of spermatogenesis and mature

  15. PCR-based molecular characterization and insilico analysis of food-borne trematode parasites Paragonimus westermani, Fasciolopsis buski and Fasciola gigantica from Northeast India using ITS2 rDNA

    PubMed Central

    Prasad, Pramod Kumar; Goswami, Lalit mohan; Tandon, Veena; Chatterjee, Anupam

    2011-01-01

    Food-borne fluke infections/trematodiases are emerging as a major public health problem worldwide with over 40 million people affected and over 10% of world population at risk of infection. The major concentration of these infections is in Southeast Asian and Western Pacific Regions, where the epidemiological factors (including the prevalent socio-cultural food habits) are conducive for transmission of these infections. The preponderance of these infections is usually in food deficit poor communities that lack access to proper sanitary infrastructure. While targeting health for all, especially the poor rural tribal communities, it is imperative to take these infections into account. Bayesian analysis phylogeny of food-borne trematode parasites under study showed that they are closely related phylogenetic groups. To focus the control strategies at the target populations, the aim of the present study was to establish molecular methods for accurate discrimination between common food-borne trematodes parasites Paragonimus (lung fluke), Fasciolopsis (giant intestinal fluke) and Fasciola (liver fluke), the infections of which commonly prevail in NE India. In the first step, we amplified and sequenced the second internal transcribed spacer (ITS2) region of ribosomal DNA, utilizing nucleotide differences between the multiple sequence alignments of the parasites under study. Based upon the differences in nucleotide sequences of conserved regions, we designed species-specific primers that can unequivocally discriminate one species from another. ITS2 sequence motifs allowed an accurate in-silico distinction of the trematodes. The data indicate that ITS2 motifs (≤ 50 bp in size) can be considered promising tool for trematode species identification. Using molecular morphometrics that is based on ITS2 secondary structure homologies, phylogenetic relationships with various isolates of several trematode species have been discussed. The present results suggest that the ITS2

  16. Fasciola hepatica: Specificity of a coproantigen ELISA test for diagnosis of fasciolosis in faecal samples from cattle and sheep concurrently infected with gastrointestinal nematodes, coccidians and/or rumen flukes (paramphistomes), under field conditions.

    PubMed

    Kajugu, P-E; Hanna, R E B; Edgar, H W; McMahon, C; Cooper, M; Gordon, A; Barley, J P; Malone, F E; Brennan, G P; Fairweather, I

    2015-09-15

    Chronic fasciolosis is often diagnosed by faecal egg counting (FEC), following concentration of the eggs in the sample by a zinc sulphate floatation method. However, concentration by a sedimentation technique gives improved sensitivity. Interpretation of FEC results for fasciolosis is complicated by factors such as the long pre-patent period and irregular egg shedding. Thus, FEC reduction tests (FECRT), when used alone, are not completely reliable for diagnosis of anthelmintic susceptibility or resistance in local fluke populations, especially when parasite burdens are small. A Fasciola hepatica coproantigen ELISA test has been introduced which more accurately reflects the presence of flukes in the host bile ducts in late pre-patent infections, and absence of flukes following successful chemotherapeutic intervention. The aim of the present study was to elucidate the specificity of the F. hepatica coproantigen ELISA technique, particularly regarding potential cross-reactivity with rumen fluke (paramphistome), gastrointestinal nematode and coccidian infections. The method involved parallel testing of a large battery of faecal samples from field-infected cattle and sheep using floatation and sedimentation FECs and coproantigen analysis. No evidence was found for significant false positivity in the F. hepatica coproantigen ELISA due to paramphistome, coccidian and/or gastrointestinal nematode co-infections. With sedimentation FECs less than 10 F. hepatica eggs per gram (epg), the likelihood of a positive coproantigen result for the sample progressively decreased. Diagnosis of fasciolosis should be based on consideration of both FEC and coproantigen ELISA findings, to ensure optimum sensitivity for pre-patent and low-level infections.

  17. Fasciola hepatica: a comparative survey of adult fluke resistance to triclabendazole, nitroxynil and closantel on selected upland and lowland sheep farms in Northern Ireland using faecal egg counting, coproantigen ELISA testing and fluke histology.

    PubMed

    Hanna, R E B; McMahon, C; Ellison, S; Edgar, H W; Kajugu, P-E; Gordon, A; Irwin, D; Barley, J P; Malone, F E; Brennan, G P; Fairweather, I

    2015-01-15

    In order to investigate the incidence and distribution of adult fluke resistance to the fasciolicide tricalbendazole (TCBZ) amongst populations of Fasciola hepatica in sheep flocks in Northern Ireland (NI), individual rectal faeces samples were collected from 3 groups of 20 sheep, before (pre-dose), and 21 days after (post-dose) treatment of the animals with TCBZ, nitroxynil or closantel, on each of 13 well-managed sheep farms distributed across the province. The efficacy of each flukicide was determined for each farm, using faecal egg count reduction (FECRT) and F. hepatica coproantigen ELISA testing. In certain flocks, 2 sheep with high pre-dose faecal egg counts (FEC) were killed 3 days and 21 days respectively after TCBZ treatment, and the histology of the fluke reproductive organs was compared with that of flukes from untreated sheep, and from sheep treated with nitroxynil or closantel 2 days prior to death, using haematoxylin and eosin (H&E) staining and an in situ hybridisation method (TdT-mediated dUDP nick end labelling [TUNEL]) to demonstrate apoptosis. Results from FECRT revealed that in all flocks with a high fluke burden, TCBZ was ineffective in treating chronic fasciolosis, and this finding was generally supported by the results of the coproantigen reduction test (CRT). The histology of reproductive organs of flukes from TCBZ-treated sheep in these flocks was normal, when compared with untreated flukes, and this, together with the FECRT and CRT findings, indicated a likely diagnosis of TCBZ resistance in all the flocks with a high fluke burden. In contrast, nitroxynil and closantel were found to be fully effective against TCBZ-resistant flukes in each of the flocks bearing a high chronic fluke burden. All of the flocks with a high fluke burden and TCBZ resistance were managed on lowland in the South and East of NI. Upland flocks, in the North and West, had low fluke burdens, or were clear of infection; and FECs were too low to allow valid resistance

  18. A checklist of cercariae (Trematoda: Digenea) in molluscs from Brazil.

    PubMed

    Pinto, Hudson Alves; De Melo, Alan Lane

    2013-01-01

    A checklist of digenetic trematodes found in molluscs from Brazil is presented based on 127 scientific articles published after a century of studies. To date 23 families, 35 genera and 46 species of trematodes were identified infecting 25 species of molluscs in the country. Another 36 species described in the collective-group Cercaria were found in 15 species of molluscs and have not yet been associated with the respective adult parasites. Larvae found in 20 species of molluscs and grouped into 10 cercarian types are also listed.

  19. [Trematodes (Trematoda) of bats (Chiroptera) from the Middle Volga Region].

    PubMed

    Kirillov, A A; Kirillova, N Iu; Vekhnik, V P

    2012-01-01

    The data on species diversity of trematodes from bats collected in the Middle Volga Region are summarized. According to original and literary data, 20 trematode species were recorded in bats of the region examined. Plagiorchis elegans, Lecithodendrium skrjabini, L. rysavyi, Prosthodendrium hurkovaae, and Pycnoporus megacotyle are specified for the bat fauna of Russia for the first time. For 11 species of parasites, new hosts are recorded. The analysis of bat helminthes demonstrated that the fauna of trematodes of the northern bat (12 species of trematodes), of the pond, and of the Brandt's bats is the most diverse, constituting more than 10 parasite species per bat species. The largest number of final hosts in the Middle Volga Region is characteristic of Plagiorchis koreanus and Prosthodendrium chilostomum; the latter species were revealed in 8 and 7 bat species, respectively. Trematodes of bats possess a high degree of host specificity. 17 species parasitize exclusively in bats out of 20 parasite species registered for the order Chiroptera. Only 3 species (Plagiorchis elegans, P. vespertilionis, and Prosthodendrium chilostomum) show wide degree of specificity, being found in other animals. Taxonomic position, the circle of hosts, collecting sites, and brief data in biology and geographical distribution for each helminth species are specified. Morphological descriptions and original figures for all the trematode species revealed in bats of the Middle Volga Region are given.

  20. Nilocotyle duplicisphinctris sp. n. (Trematoda: Paramphistomidae) from Hippopotamus amphibius.

    PubMed

    Sey, O; Graber, M

    1980-06-01

    Nilocotyle duplicisphinctris sp. n. was recovered from the stomach of Hippopotamus amphibius in Ethiopia. The average body size (L x W) is 3.6 x 1.2 mm; pharynx 0.5 x 0.3, Dicranocoelium type; outer diameter of the acetabulum 0.85, Nilocotyle type; anterior testis 0.34 x 0.49; posterior testis 0.32% x 0.27: ovary 0.27 x 0.19; genital opening 0.30 x 0.34, Duplicisphinctris type and eggs 0.15 x 0.07. The most characteristic specific feature is the presence of a double sphincter along the genital atrium.

  1. Life history of Echinoparyphium recurvatum (Trematoda: Echinostomatidae) in Korea.

    PubMed

    Sohn, W M

    1998-06-01

    The present study was performed to observe characteristics of the life history of Echinoparyphium recurvatum under both natural and laboratory conditions in Korea. A batch of Radix auricularia coreana was collected from Sunamchon, one of the stream of West Naktonggang (River), in Kangso-gu, Pusan during August and September 1992. Out of 106 snails examined by crushing, 52 (49.0%) were infected with larval E. recurvatum, i.e. rediae, cercariae and metacercariae. Cercariae naturally shed from snails encysted in the snails of same species and loaches, but not in mud-snails. Adult worms were detected from chicks and ducks experimentally infected with metacercariae, but not from rats and mice. The average recovery rate of adults from chicks was 13.1%. Rediae were sac-like, 2.437 x 0.317 mm in average size, with a muscular pharynx and a brownish cecum which reached the anterior half of the body. Cercariae consisted of a spindle-shaped body (0.262 x 0.129 mm in average) and a rod-like tail (0.528 x 0.056 mm in average). In the cercarial body, 45 collar spines were observed on the head crown, and double rows of excretory ducts with fine granules were laterally arranged between the pharynx and the ventral sucker. Metacercariae were spherical, 0.144 x 0.142 mm in average size, with thick hyaline outer and thin elastic inner walls, and many excretory granules. Adults were slender and more attenuated in the anterior end, 2.760 x 0.550 mm in average size, and had 45 collar spines including four end group spines on both ventral corners. From the above results, it was confirmed that R. auricularia coreana plays a pivotal role in the life cycle of E. recurvatum as the first and/or second intermediate hosts in Korea. PMID:9637826

  2. Egg and miracidium of Hirudinella ventricosa (Trematoda: Hirudinellidae).

    PubMed

    Murugesh, M; Madhavi, R

    1990-10-01

    It is reported for the first time that Hirudinella ventricosa releases eggs in strings, each string containing active spermatozoa and numerous oval, thick-shelled, translucent eggs measuring 40-42 x 28-30 microns. Each egg contained a fully developed miracidium with an anterior crown of spines.

  3. [Muscular system of the Microphallus pygmaeus metacercaria (Trematoda: microphallidae)].

    PubMed

    Krupenko, D Iu

    2010-01-01

    The musculature of body wall, attachment organs (oral and ventral suckers), and internal organs (alimentary tract, excretory bladder, and ducts of reproductive system) of the Microphallus pygmaeus metacercaria are described. An unusual arrangement of longitude and diagonal muscles was found in the hind part of the metacercaria body. Longitude fibers of dorsal and lateral body surfaces are bow-shaped and bend round the excretory pore along its ventral margin. An additional group of diagonal fibers is situated in the hind part of ventral body surface.

  4. A Ribeiroia spp. (Class: Trematoda) - Specific PCR-based diagnostic

    USGS Publications Warehouse

    Reinitz, D.M.; Yoshino, T.P.; Cole, R.A.

    2007-01-01

    Increased reporting of amphibian malformations in North America has been noted with concern in light of reports that amphibian numbers and species are declining worldwide. Ribeiroia ondatrae has been shown to cause a variety of types of malformations in amphibians. However, little is known about the prevalence of R. ondatrae in North America. To aid in conducting field studies of Ribeiroia spp., we have developed a polymerase chain reaction (PCR)-based diagnostic. Herein, we describe the development of an accurate, rapid, simple, and cost-effective diagnostic for detection of Ribeiroia spp. infection in snails (Planorbella trivolvis). Candidate oligonucleotide primers for PCR were designed via DNA sequence analyses of multiple ribosomal internal transcribed spacer-2 regions from Ribeiroia spp. and Echinostoma spp. Comparison of consensus sequences determined from both genera identified areas of sequence potentially unique to Ribeiroia spp. The PCR reliably produced a diagnostic 290-base pair (bp) product in the presence of a wide concentration range of snail or frog DNA. Sensitivity was examined with DNA extracted from single R. ondatrae cercaria. The single-tube PCR could routinely detect less than 1 cercariae equivalent, because DNA isolated from a single cercaria could be diluted at least 1:50 and still yield a positive result via gel electrophoresis. An even more sensitive nested PCR also was developed that routinely detected 100 fg of the 290-bp fragment. The assay did not detect furcocercous cercariae of certain Schistosomatidae, Echinostoma sp., or Sphaeridiotrema globulus nor adults of Clinostomum sp. or Cyathocotyle bushiensis. Field testing of 137 P. trivolvis identified 3 positives with no overt environmental cross-reactivity, and results concurred with microscopic examinations in all cases. ?? American Society of Parasitologists 2007.

  5. Identification and expression of Fasciola gigantica thioredoxin.

    PubMed

    Changklungmoa, Narin; Kueakhai, Pornanan; Apisawetakan, Somjai; Riengrojpitak, Suda; Sobhon, Prasert; Chaithirayanon, Kulathida

    2014-06-01

    In the present study, a cDNA encoding Trx from F. gigantica (FgTrx) was cloned by polymerase chain reaction (PCR). The sequence of FgTrx, analyzed by BLAST, SignalP, and ClustralW programs, showed 315 bp of an open reading frame (ORF), 12 bp 5'UTR, 78 bp 3'UTR, and the putative FgTrx peptide comprising of 104 amino acids, with a molecular weight of 11.68 kDa, with the active site containing five amino acids (tryptophan, cysteine, glycine, proline, cysteine) with a conserved dithiol motif from the two cysteines, and pI 5.86. The peptide had no signal sequence; hence, it was not a secreted protein. The recombinant FgTrx was expressed in Escherichia coli BL21 (DE3) and used for production for a polyclonal antibody in rabbits (anti-rFgTrx). The FgTrx protein expression, estimated by indirect ELISA using the rabbit anti-rFgTrx as probe, showed high levels in eggs, 2- and 4-week-old juveniles, and adult parasite. In a functional test, the rFgTrx exhibited specific activity that could be suppressed by an inhibitor (PX12). When tested by immunoblotting and immunohistochemistry, rabbit anti-rFgTrx reacted with natural FgTrx at a molecular weight of 11.68 kDa from eggs, metacercariae, NEJ, 2- and 4-week-old juveniles, and adult F. gigantica. The FgTrx protein was distributed at high levels in the tegument of 2- and 4-week-old juveniles, and the tegument, parenchyma, eggs, and reproductive organs of adult parasites. FgTrx 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 drug target. PMID:24718754

  6. Fasciola hepatica: Histology of the Reproductive Organs and Differential Effects of Triclabendazole on Drug-Sensitive and Drug-Resistant Fluke Isolates and on Flukes from Selected Field Cases.

    PubMed

    Hanna, Robert

    2015-01-01

    This review summarises the findings of a series of studies in which the histological changes, induced in the reproductive system of Fasciola hepatica following treatment of the ovine host with the anthelmintic triclabendazole (TCBZ), were examined. A detailed description of the normal macroscopic arrangement and histological features of the testes, ovary, vitelline tissue, Mehlis' gland and uterus is provided to aid recognition of the drug-induced lesions, and to provide a basic model to inform similar toxicological studies on F. hepatica in the future. The production of spermatozoa and egg components represents the main energy consuming activity of the adult fluke. Thus the reproductive organs, with their high turnover of cells and secretory products, are uniquely sensitive to metabolic inhibition and sub-cellular disorganisation induced by extraneous toxic compounds. The flukes chosen for study were derived from TCBZ-sensitive (TCBZ-S) and TCBZ-resistant (TCBZ-R) isolates, the status of which had previously been proven in controlled clinical trials. For comparison, flukes collected from flocks where TCBZ resistance had been diagnosed by coprological methods, and from a dairy farm with no history of TCBZ use, were also examined. The macroscopic arrangement of the reproductive system in flukes was studied using catechol/carmine stained whole mounts, and the histology of the main organs was examined using conventional haematoxylin-eosin stained sections. Validation of apoptosis in the fluke sections was carried out using an in situ hybridisation method designed to label endonuclease-induced DNA strand breaks. In TCBZ-S flukes exposed to TCBZ metabolites for 24-96 h in vivo, but not in TCBZ-R flukes, those tissues where active meiosis and/or mitosis occurred (testis, ovary, and vitelline follicles), were found to display progressive loss of cell content. This was due to apparent failure of cell division to keep pace with expulsion of the mature or effete products

  7. Fasciola hepatica: Histology of the Reproductive Organs and Differential Effects of Triclabendazole on Drug-Sensitive and Drug-Resistant Fluke Isolates and on Flukes from Selected Field Cases

    PubMed Central

    Hanna, Robert

    2015-01-01

    This review summarises the findings of a series of studies in which the histological changes, induced in the reproductive system of Fasciola hepatica following treatment of the ovine host with the anthelmintic triclabendazole (TCBZ), were examined. A detailed description of the normal macroscopic arrangement and histological features of the testes, ovary, vitelline tissue, Mehlis’ gland and uterus is provided to aid recognition of the drug-induced lesions, and to provide a basic model to inform similar toxicological studies on F. hepatica in the future. The production of spermatozoa and egg components represents the main energy consuming activity of the adult fluke. Thus the reproductive organs, with their high turnover of cells and secretory products, are uniquely sensitive to metabolic inhibition and sub-cellular disorganisation induced by extraneous toxic compounds. The flukes chosen for study were derived from TCBZ-sensitive (TCBZ-S) and TCBZ-resistant (TCBZ-R) isolates, the status of which had previously been proven in controlled clinical trials. For comparison, flukes collected from flocks where TCBZ resistance had been diagnosed by coprological methods, and from a dairy farm with no history of TCBZ use, were also examined. The macroscopic arrangement of the reproductive system in flukes was studied using catechol/carmine stained whole mounts, and the histology of the main organs was examined using conventional haematoxylin-eosin stained sections. Validation of apoptosis in the fluke sections was carried out using an in situ hybridisation method designed to label endonuclease-induced DNA strand breaks. In TCBZ-S flukes exposed to TCBZ metabolites for 24–96 h in vivo, but not in TCBZ-R flukes, those tissues where active meiosis and/or mitosis occurred (testis, ovary, and vitelline follicles), were found to display progressive loss of cell content. This was due to apparent failure of cell division to keep pace with expulsion of the mature or effete products

  8. Pumilio genes from the Platyhelminthes.

    PubMed

    Koziol, Uriel; Marín, Monica; Castillo, Estela

    2008-01-01

    Pumilio proteins are proposed to have a conserved primordial function in the maintenance of proliferation in stem cells through post-transcriptional regulation. In this work, a search for pumilio homology domain (PUM-HD) sequences of pumilio genes from several Platyhelminthes species was performed, including representatives form Cestoda, Trematoda and Tricladida. Only one PUM-HD sequence was found in each triclad species; however, two PUM-HD homologues were found in all the parasitic species. These sequences formed two clearly separated clades: PlatyPum1, with sequences from all species, and PlatyPum2, composed exclusively of neodermatan sequences. Therefore, at least one duplication of the pumilio gene must have occurred before the divergence of cestodes and trematodes. Further duplications of PUM-HD were found in Fasciola hepatica, but these consist of retropseudogenes. This is the first comparative analysis of PUM-HD sequences in the Platyhelminthes and, more generally, in any lophotrochozoan phylum.

  9. Experimentally induced Fasciola hepatica infections in black-tailed deer.

    PubMed

    Kistner, T P; Koller, L D

    1975-04-01

    The susceptibility of black-tailed deer (Odocoileus hemionus columbianus) to the common liver fluke (F. hepatica) was studied. Two deer and one sheep comprised each of three experimental groups. Animals in each group were inoculated individually with 250, 500, or 1000 F. hepatica metacercariae. One deer and one sheep given 1000 metacercariae died with lesions consistent with black disease 7 weeks after inoculation. At necropsy 6 or 15 weeks postinoculation, the mean percentage recovery of the inoculum was 38.9% from the deer and 51.9% from the sheep. Fluke eggs recovered from the deer were viable and metacercariae cultured from the eggs were fully infective for sheep. Pathologic changes associated with F. hepatica infection were more severe in the infected deer; consequently, the deer were less resistant to the lethal effects of the parasite than sheep. Considering the experimental results and the fact that naturally acquired common liver fluke infection has been reported infrequently from black-tailed deer, it was concluded that black-tailed deer do not constitute a significant reservoir for F. hepatica in domestic livestock.

  10. Morphological and Molecular Characterization of Clinostomum detruncatum (Trematoda: Clinostomidae) Metacercariae Infecting Synbranchus marmoratus.

    PubMed

    Acosta, Aline Angelina; Caffara, Monica; Fioravanti, Maria Letizia; Utsunomia, Ricardo; Zago, Aline Cristina; Franceschini, Lidiane; da Silva, Reinaldo José

    2016-02-01

    A complete morphological description, supported by molecular data, of Clinostomum detruncatum metacercariae is reported in this study. The metacercariae were found infecting Synbranchus marmoratus from the Paraná River, municipality of Guaíra, Paraná State, Brazil. The taenioidean uterus is the main morphological characteristic of this species. Both neighbor-joining (NJ) and maximum likelihood (ML) trees showed 2 clear separate clusters grouping together the species from the Palearctic region (Clinostomum complanatum, Clinostomum cutaneum, Clinostomum phalacrocoracis, Clinostomum philippinense) and those from the Nearctic/Neotropical regions (Clinostomum marginatum, Clinostomum tataxumui, C. detruncatum, Clinostomum sp. 7). The latter clade is divided into 2 subclades grouping species from North America and Mexico (C. marginatum and C. tataxumui), and those from Brazil (C. detruncatum and Clinostomum sp. 7). PMID:26509452

  11. Can the same species of Platynosomum (Trematoda: Dicrocoeliidae) infect both mammalian and avian hosts?

    PubMed

    Pinto, H A; Mati, V L T; Melo, A L

    2016-05-01

    The importance of platynosomiasis has increased in feline veterinary practice, but aspects related to the specificity of Platynosomum spp. in definitive hosts requires further study. Although morphological traits suggest that the same species, P. illiciens, may infect both birds and mammals, the synonymies previously proposed have not been widely accepted, likely because host specificity is assumed. In addition, the name P. fastosum has frequently been used for parasites recovered from mammals. In the present study, metacercariae (n= 100/animal) of P. illiciens recovered from lizards (Hemidactylus mabouia) in Brazil were fed to Australian parakeets (Melopsittacus undulatus) and mice. Two parasites were recovered from the liver of one M. undulatus specimen during a necropsy that was performed 105 days after infection, and all mice were found to be infected with 37 ± 12 (18-48) parasites. The morphology of the P. illiciens obtained from the parakeet was similar to that of parasites obtained from mice and those described previously from naturally infected birds and mammals. Non-specificity of P. illiciens in hosts is discussed briefly, based on the parasitological and morphological results obtained during the avian experimental platynosomiasis and the epidemiology and geographical distribution of this parasite. PMID:25781630

  12. [Variations in the number of metacercariae in the daughter sporocysts of Microphallus pygmaeus (Trematoda: Microphallidae)].

    PubMed

    Galaktionov, K V

    1980-01-01

    The daughter sporocystes of Microphallus pygmaeus from the snails of Littorina saxatilis and L. obtusata were investigated. The average quantity of metacercariae in sporocysts is influenced by host species, its syze and by the season. There are differences in the average number of larvae from parthenites of L. saxatilis, collected in the Barents and White seas. PMID:6445532

  13. [Genetic heterogeneity in natural populations of Microphallus piriformes and M. pygmaeus parthenites (Trematoda: Microphallidae)].

    PubMed

    Khalturin, K V; Mikhaĭlova, N A; Granovich, A I

    2000-01-01

    The random amplified polymorphic DNA (RAPD) technique was applied for the studies of genetic heterogeny between several natural populations of trematodes belonging to the Microphallida family. Initially, the metacercariae from the daughter sporocysts infestating Littorina saxatilis and Littorina littorea periwinkles were used. Comparison of the banding patterns obtained for the different metacercariae within one sporocyst gave an unpredicted results. For two of three studied species (M. pygmaeus and M. pseudopygmaeus), the considerable differences in RAPD patterning was detected. According to the classical point of view, the process of cercariae (metacercariae in case of the "pygmaeus" group of microphallides) formation does not include DNA recombination. Because of that, all metacercariae within one single sporocyst should be genetically identical. However, data obtained clearly shows that at least in some cases it is not so. We can hardly believe that such result could be a methodological artifact, for not single difference in a RAPD patterns was recorded between the metacercariae within sporocysts of M. piriformes. Moreover, even the 100 fold dilution of the total DNA used for PCR amplification does not change the banding patterns. Hence, our results can not be explained by slight fluctuations in the DNA concentrations between the samples. The most evident conclusion is that we came across yet strange, but real phenomenon--some degree of genetic difference within the progeny of each of the sporocysts--metacercariae. However, the detailed study is needed to understand and interpret these observations correctly. Amplification of the total DNA extracted from the whole sporocysts (containing metacercariae) never showed any differences in RAPD patterns between the parasites been derived from one infestated snail (local parasite hemipopulation). That allowed us to compare different parasite populations referring the RAPD pattern of one sporocyst from a snail to as a representative of one local hemipopulation. Analysis of the RAPD-loci frequencies showed a considerable genetic differences between the subpopulations of M. piriformes, infestating different paraxenic intermediate hosts--L. saxatilis and L. obtusata. This phenomenon was statistically significant for 2 localities of 3 studied. No heterogeny within populations was recorded for M. pygmaeus. Both M. piriformes and M. pygmaeus are characterized by the genetic differentiation in the microgeographic scale (within the Chupa bay of the White sea, the longest distance between the analyzed localities is 20 km). According to the frequencies of the RAPD-loci, parasites from the sheltered locality differ significantly from the parasites of other two localities exposed to the open sea. For both species the degree of genetic similarity between the populations correlates positively with the distance between the localities. We can suppose that the population structure of microphallids depended mainly upon the population structures of their intermediate hosts, definitive hosts and geographical structure of the areal. However, taking into consideration the low motility of snails, we believe that the distribution, migration and species composition of the definitive hosts play the key role in the genetic structuring of M. pygmaeus and M. piriformes hemipopulations. As an addition, the RAPD analysis of the parasite populations from the Barents Sea (East cost of Murman peninsula) and North Sea (Western cost of Sweden) revealed no significant genetic differences between the worms from those places and from the White Sea. However in case of this macrogeographic comparison, insufficient number of samples does not allow us to draw any final conclusions. PMID:11212616

  14. Developmental and Phylogenetic Characteristics of Stellantchasmus falcatus (Trematoda: Heterophyidae) from Thailand

    PubMed Central

    Sripalwit, Pralongyut; Wongsawad, Chalobol; Chontananarth, Thapana; Anuntalabhochai, Somboon; Wongsawad, Pheravut; Chai, Jong-Yil

    2015-01-01

    This study aimed to investigate the infection status, worm development, and phylogenetic characteristics of the intestinal trematode, Stellantchasmus falcatus. The metacercariae of S. falcatus were detected only in the half-beak (Dermogenus pusillus) out of the 4 fish species examined. Their prevalence was 90.0%, and the intensity of infection was 919 metacercariae on average. Worms were recovered from 33 (97.1%) of 34 chicks that were experimentally infected with 200 S. falcatus metacercariae each, and the average recovery rate was 43.0%. The body size and inner organs of S. falcatus quickly increased in the experimental chicks over days 1-2 post-infection (PI). In addition, ITS2 sequence data of this parasite were analyzed to examine the phylogenetic relationships with other trematodes using the UPGMA method. The results indicated that the ITS2 sequence data recorded from trematodes in the family Heterophyidae appeared to be monophyletic. This study concluded that D. pusillus serves as a compatible second intermediate host of S. falcatus in Thailand and that S. falcatus can develop rapidly in the experimental chicks. Data collected from this study can help to close the gap in knowledge regarding the epidemiology, biology, and phylogenetic characteristics of S. falcatus in Thailand. PMID:25925179

  15. [Effectiveness of oxyclozanide in cattle naturally invaded by Liorchis scotiae trematoda].

    PubMed

    Corba, J; Pacenovský, J; Krupicer, I; Breza, M; Popovic, S; Reisz, T

    1976-01-01

    The paper reports on the results of the clinical tests for the effectiveness of oxyclozanide pure substance (produced by ICI, U.K.) in cattle naturally invaded by the paramphistomata Liorchis scotiae. Helminthological dissection, performed 21 days after a single application of 15 mg oxyclozanide per 1 kg 1. w., revealed 87.5% intenseffectiveness on sexually mature paramphistomata on 85% intenseffectiveness on juvenile paramphistomata. The extenseffectiveness of the chemical was equal to zero. The coprological examinations performed for the three weeks in one-week intervals after therapy showed a decline in the number of produced eggs. The animals were given the medicated feed containing oxyclozanide only after preceding starvation. The chemical did not produce any unfavourable side effects or signs.

  16. Two new species of Phyllodistomum Braun, 1899 (Trematoda: Gorgoderidae Looss, 1899) from Great Barrier Reef fishes.

    PubMed

    Ho, Hei Wa; Bray, Rodney A; Cutmore, Scott C; Ward, Selina; Cribb, Thomas H

    2014-01-01

    Two new species of Phyllodistomum Braun, 1899 are described from the urinary bladder of fishes of the Great Barrier Reef. Phyllodistomum hoggettae n. sp. is described from Plectropomus leopardus (leopard coralgrouper) (Serranidae) and P. vaili n. sp. is described from Mulloidichthys vanicolensis (yellowfin goatfish) and M. flavolineatus (yellowstripe goatfish) (Mullidae). These species are compared with 26 previously described marine Phyllodistomum species and found to be distinct in combinations of body shape, sucker ratio and shape of the gonads. Preliminary molecular data also demonstrate that they are distinct from each other and for those other species for which data are available. PMID:24871750

  17. Morphological and histological identification of Paramphistomum cervi (Trematoda: Paramiphistoma) in the rumen of infected sheep

    PubMed Central

    Chaoudhary, Vijayata; Hasnani, J. J.; Khyalia, Mukesh K.; Pandey, Sunanda; Chauhan, Vandip D.; Pandya, Suchit S.; Patel, P. V.

    2015-01-01

    Aim: This study was undertaken to identify Paramphistomum cervi on the basis of its morphology and histology to be the common cause of paramphistomosis in infected sheep and its differentiation from other similar Paramphistomes in Gujarat. Materials and Methods: Adult rumen flukes were recovered from the rumen of naturally infected sheep slaughtered in various abattoirs in Gujarat. Some adult flukes were flattened and stained in Borax carmine, and some were sectioned in the median sagittal plane and histological slides of the flukes were prepared for detailed morphological and histological studies. Result: Microscopic pictures of the parasite used in identification define the similarity in the morphology and histology of the anterior sucker, pharynx, esophagus, genital atrium, posterior sucker (acetabulum) and testes to the P. cervi. Conclusion: It can be concluded that the most common species found in sheep infected with Paramphistomosis is P. cervi on the basis of its histo-morphological appearance in Gujarat. PMID:27047009

  18. Novel Infection Site and Ecology of Cryptic Didymocystis sp. (Trematoda) in the Fish Scomberomorus maculatus.

    PubMed

    Schrandt, Meagan N; Andres, Michael J; Powers, Sean P; Overstreet, Robin M

    2016-06-01

    An undescribed, cryptic species of Didymocystis, as determined from sequences of 2 ribosomal genes and superficially similar to Didymocystis scomberomori ( MacCallum and MacCallum, 1916 ), infected the skin of the Spanish mackerel, Scomberomorus maculatus , in the north-central Gulf of Mexico (GOM). An analysis of 558 fish from 2011 to 2013 from Louisiana, Mississippi, Alabama, and the Florida panhandle showed the prevalence of the trematode varied both spatially and temporally but not with sex of the fish host. Month, year, and geographic location were identified by a negative binomial generalized linear model as indicators of the abundance and intensity of infection. Prevalence, abundance, and intensity of infection were greatest in spring and fall months off the Florida panhandle. Furthermore, the abundance and intensity of infection correlated negatively with fork length, weight, and gonad weight of mature fish but positively with longitude. Therefore, smaller adult fish tended to be more infected than larger adults, and prevalence and intensity increased from west to east (Louisiana to Florida). Spatial and temporal trends seemed to result from physical factors (e.g., water temperature, salinity, bottom type), but they also coincided with the annual migration of S. maculatus as fish moved northward along the GOM coastline from the southern tip of Florida in the spring months and returned in the fall, being present in the north-central GOM from late spring through fall. This pattern suggests the possibility that acquisition of infections occurred from a molluscan host in waters off the Florida panhandle. PMID:26981762

  19. Toxicity of cadmium and zinc mixtures to cercarial tail loss in Diplostomum spathaceum (Trematoda:Diplostomidae).

    PubMed

    Morley, N J; Crane, M; Lewis, J W

    2005-01-01

    The effects of cadmium and zinc mixtures on tail loss of the free-living cercarial stage of the parasitic fluke Diplostomum spathaceum were investigated at concentrations ranging from 0.1 to 10,000 microg/L. Cercariae were exposed to metal mixtures of equal concentrations, metal mixtures of unequal concentrations, and a low-dose pretreatment followed by high-dose exposure mixtures. Under control conditions as the cercarial population aged tail loss occurred naturally a few hours before death, with an increasing tail loss over time associated with a decrease in cercarial survival. Under all mixture exposures the period of tail loss was prolonged compared to single-metal exposures. In most test solutions this was associated with a similar increase in survival of cercariae. Inhibition of tail loss occurred in low concentrations (0.1-100 microg/L) of equal mixed exposures. In some unequal, and in all pretreatment mixture exposures, a significant stimulation of tail loss occurred compared to the pattern of decreasing survival over time within the cercarial population. The importance of toxic metal mixtures to cercarial tail loss and potential implications for successful parasite transmission are discussed. PMID:15482840

  20. Life history and biology of Fascioloides magna (Trematoda) and its native and exotic hosts

    PubMed Central

    Malcicka, Miriama

    2015-01-01

    Host–parasite interactions are model systems in a wide range of ecological and evolutionary fields and may be utilized for testing numerous theories and hypotheses in terms of both applied and fundamental research. For instance, they are important in terms of studying coevolutionary arms races, species invasions, and in economic terms the health of livestock and humans. Here, I present a comprehensive description of the life history, biogeography, and biology of the giant liver fluke, Fascioloides magna, and both its intermediate and definitive hosts. F. magna is native to North America where it uses several species of freshwater snails (Lymnaeidae) as intermediate hosts and four main species of ungulates as definitive hosts. The fluke has also been introduced into parts of Europe where it is now established in two lymnaeid snail species and three ungulate species. This study gives a comprehensive description of different developmental stages of the fluke in its two host classes, as well as detailed notes on historical and present distributions of F. magna in North America and Europe as well as in its snail and deer hosts (with range maps provided). Aberrant and dead-end hosts are also discussed in detail, and descriptive phylogenies are provided for all of the organisms. I briefly discuss how F. magna represents a model example of multiple-level ecological fitting, a phenomenon not yet described in the empirical literature. Lastly, I explore possible future scenarios for fluke invasion in Europe, where it is currently expanding its range. PMID:25897378

  1. Metacercariae of Clinostomum complanatum (Trematoda: Digenea) in European newts Triturus carnifex and Lissotriton vulgaris (Caudata: Salamandridae).

    PubMed

    Caffara, M; Bruni, G; Paoletti, C; Gustinelli, A; Fioravanti, M L

    2014-09-01

    Adults of Clinostomum spp. are digenetic trematodes found in fish-eating birds, reptiles and occasionally mammals, including humans. Freshwater snails serve as first intermediate hosts and many fish species and amphibians as second intermediate hosts. To date, amphibian hosts of Clinostomum metacercariae include members of urodele and anuran families in North America, but no data are available on infections of European amphibians, including newts. In this study, we characterize infections of Clinostomum complanatum metacercariae in four smooth (Lissotriton vulgaris) and 18 Italian crested newts (Triturus carnifex) from an artificial pond located in a protected area in Tuscany, Italy. Parasites were surgically removed from the infected newts and identified both morphologically and using sequences of a mitochondrial gene, cytochrome c oxidase I, and the ribosomal markers, internal transcribed spacers. This is the first record of C. complanatum in European newts and, more generally, in amphibians in Europe.

  2. Experimental centrocestiasis: Worm burden, morphology and fecundity of Centrocestus formosanus (Trematoda: Heterophyidae) in dexamethasone immunosuppressed mice.

    PubMed

    Pinto, Hudson Alves; Mati, Vitor Luís Tenório; de Melo, Alan Lane

    2015-10-01

    Centrocestus formosanus is an intestinal foodborne trematode with medical and veterinary importance that remains with the pathological and immunological aspects of the infection in definitive host poorly studied. In the present study, we evaluated the effects of pharmacological immunosuppression by glucocorticoids in experimental centrocestiasis. Mice of the AKR/J strain were orally inoculated with 100 metacercariae of C. formosanus obtained in naturally infected fish (Australoheros facetus) collected in an urban reservoir from Brazil. Treatment with dexamethasone (25 mg/kg, via subcutaneous injection) was started 1h before infection of mice and then continued daily during 14 days post-infection. Untreated mice also infected with C. formosanus were used as control. At the end of the treatment course, all rodents were euthanized and adult parasites recovered from host intestines were subjected to morphological and morphometric analysis under optical microscopy. The worm burden in dexamethasone treated group [70±14 (41-85)] was significantly greater (p<0.0001) than that in the control group [15±4 (10-22)]. In addition, the parasites recovered from immunosuppressed mice were larger, with more developed reproductive structures and greater number of intrauterine eggs than in control mice. These parasite developmental changes induced by dexamethasone treatment are reported for the first time in experimental centrocestiasis. Moreover the higher parasite fecundity induced by glucocorticoid treatment had so far not been reported for any heterophyid species, which can have implications for the pathology and morbidity in infections caused by these parasites.

  3. Can the same species of Platynosomum (Trematoda: Dicrocoeliidae) infect both mammalian and avian hosts?

    PubMed

    Pinto, H A; Mati, V L T; Melo, A L

    2016-05-01

    The importance of platynosomiasis has increased in feline veterinary practice, but aspects related to the specificity of Platynosomum spp. in definitive hosts requires further study. Although morphological traits suggest that the same species, P. illiciens, may infect both birds and mammals, the synonymies previously proposed have not been widely accepted, likely because host specificity is assumed. In addition, the name P. fastosum has frequently been used for parasites recovered from mammals. In the present study, metacercariae (n= 100/animal) of P. illiciens recovered from lizards (Hemidactylus mabouia) in Brazil were fed to Australian parakeets (Melopsittacus undulatus) and mice. Two parasites were recovered from the liver of one M. undulatus specimen during a necropsy that was performed 105 days after infection, and all mice were found to be infected with 37 ± 12 (18-48) parasites. The morphology of the P. illiciens obtained from the parakeet was similar to that of parasites obtained from mice and those described previously from naturally infected birds and mammals. Non-specificity of P. illiciens in hosts is discussed briefly, based on the parasitological and morphological results obtained during the avian experimental platynosomiasis and the epidemiology and geographical distribution of this parasite.

  4. Detection of Paragonimus mexicanus (Trematoda) metacercariae in crabs from Oaxaca, Mexico.

    PubMed

    Vargas-Arzola, Jaime; Segura-Salvador, Aristeo; Reyes-Velasco, Leobardo; Díaz-Chiguer, Dylan L; Márquez-Navarro, Adrián; León-Avila, Gloria; Ibañez-Cervantes, Gabriela; Camacho, Alejandro D; Sánchez-Manzano, Rosa Ma; Nogueda-Torres, Benjamín

    2014-09-01

    Metacercariae of Paragonimus mexicanus were collected in crabs Tehuana guerreroensis (Rathbun, 1933) in the municipality of Putla, Oaxaca, Mexico. Metacercariae were found in 20.8% of the crabs collected, with an average of 1.9 metacercarie per crab. Stained metacercariae showed the specific characteristics of P. mexicanus by morphology and sequencing a fragment of the 28S ribosomal gene obtained by PCR. These findings reveal that T. guerreroensis is an intermediate host for P. mexicanus; this new report is relevant considering the potential risk of transmission in the states of Oaxaca and Guerrero, Mexico.

  5. Hyperparasitism by Paragordius varius (Nematomorpha: Gordiida) larva of Monostome redia (Trematoda: Digenea).

    PubMed

    Hanelt, B

    2009-02-01

    Gordiid larvae enter and encyst within a large variety of aquatic vertebrates and invertebrates. Cysts of a single species of gordiid have been found in such diverse hosts as insect larvae, snails, crustaceans, fish, and many others. One particularly surprising host report was made nearly 100 yr ago, i.e., cysts within adult trematodes. The origin of these cysts remained unclear, but it was suggested that the gordiid larvae gained entry into the adult flukes via their definitive, vertebrate host. In the present study, the finding is reported of gordiid cysts within monostome redia within a Physa gyrina snail. This is the first time gordiid cysts have been reported within redia, and this provides an alternative explanation for the presence of cysts within adult trematodes.

  6. Localization of photoreceptors in the cercariae of Proterometra macrostoma (Trematoda: Azygiidae).

    PubMed

    Rowley, Marc; Massana, Kathryn; Wier, Abigail

    2011-10-01

    The swimming behavior of the cercaria of the digenetic trematode Proterometra macrostoma changes in response to light. However, this cercaria does not possess obvious eyes or eyespots. Using behavioral assays, we were able to show that both intact and distome-removed cercariae swim significantly greater vertical distances under dim, red light than under brighter, white light. Electrophysiological experiments confirmed this result and further showed that the transverse band of the tail, known to control cercariae swimming behavior, was necessary and sufficient for the display of the light-dependent swimming behavior. Together, these data show that the distome is not required for light-dependent swimming behavior in P. macrostoma cercariae and indirectly demonstrates the presence of photoreceptors in the transverse band of the cercaria tail. PMID:21510743

  7. Integrative taxonomy of European parasitic flatworms of the genus Metorchis Looss, 1899 (Trematoda: Opisthorchiidae).

    PubMed

    Sitko, Jiljí; Bizos, Jiří; Sherrard-Smith, Eleanor; Stanton, David W G; Komorová, Petronela; Heneberg, Petr

    2016-06-01

    Metorchis spp. are flukes (Platyhelminthes: Digenea) that infect vertebrates, including humans, dogs, cats, poultry and wild game, with cyprinid freshwater fish serving as typical second intermediate hosts. In their definitive hosts, the Metorchis spp. are difficult to identify to species. We provide and analyze sequences of two nuclear (18S rDNA and ITS2) and two mitochondrial (CO1 and ND1) DNA loci of four morphologically identified European species of the Metorchis, namely Metorchis albidus, Metorchis bilis, Metorchis crassiusculus and Metorchis xanthosomus, and of another opisthorchiid, Euamphimerus pancreaticus. DNA analysis suggests that the Metorchis specimens identified morphologically as M. albidus (from Lutra lutra), M. bilis (from Phalacrocorax carbo) and M. crassiusculus (from Aquila heliaca and Buteo rufinus) represent a single species. Thus, M. albidus (Braun, 1893) Loos, 1899 and M. crassiusculus (Rudolphi, 1809) Looss, 1899 are recognized as junior subjective synonyms of M. bilis (Braun, 1790) Odening, 1962. We also provide comparative measurements of the Central European Metorchis spp., and address their tissue specificity and prevalence based on the examination of extensive bird cohort from 1962 to 2015. M. bilis and M. xanthosomus can be morphologically diagnosed by measuring the extent of genitalia relative to body length and by the size ratio of their suckers. They also differ in their core definitive hosts, with ducks (Anas, Aythya) and coots (Fulica) hosting M. xanthosomus, and cormorants (Phalacrocorax), the birds of prey (Buteo, Aquila, etc.), piscivorous mammals (Lutra, Vulpes, Ursus, etc.) and humans hosting M. bilis. Previous reports on the Metorchis spp. contain numerous suspected misidentifications. PMID:26794684

  8. [Clonal variability in expression of geo- and photoorientation in cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

    PubMed

    Prokof'ev, V V; Levakin, I A; Losev, E A; Zavirinskiĭ, Ia V; Galaktionov, K V

    2011-01-01

    The study was carried out on Himasthla elongata, a digenean common in the coastal ecosystems of the northern European seas. This species utilises intertidal prosobranchs Littorina spp. as the first intermediate host, bivalves (in the White Sea, Mytilus edulis) as the second intermediate host and gulls as the final host. The periwinkles Littorina littorea infected with H. elongata rediae (parthenogenetic generations) were sampled in the intertidal of the White Sea (66 degrees 20' N, 33 degrees 38' E) and used as the source of cercariae. Periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. Photo- and geoorientation of cercariae originating from different clones and aged 1 h and 6 h were analysed separately. It was shown that in general the larvae of each clone followed the behavioural pattern characteristics of the species (positive geoorientation and negative photoorientation). However, the degree of expression of this typical behaviour was different in different clones. An especially high variability was observed in the manifestation of geoorientation (in several clones, most larvae demonstrated negative geoorientation). Differences in the distribution of cercariae in the illumination gradient were almost equally associated with the interclonal variability and the age of the larvae. On the whole, as the age of cercariae increased, the positive geoorientation became more prominent, whereas the ratio of cercariae with the typical (negative) photoorientation decreased. Statistical analysis revealed significant differences between the cercarial clones both in the initial manifestation of geo- and photoorientation and in the changes in the character of these reactions with the larval age. Taking into account that each cercarial clone investigated had the same genotype, it seems very likely that the interclonal differences noted in this study are hereditary. Maintenance of a rather high level of genetic polymorphism by the character "expression of orientation reaction" in trematode cercariae may enhance the chances for successful transmission of these larvae. Such variability increases the scale of cercarial dispersion in space and promotes the successful infection of the hosts, whose behaviour is also subject to intra- and inter-population variability. Besides, cercariae whose behaviour deviates from the basic behaviour of the species may play the role of the population's potential for colonisation of new species of animal hosts. PMID:22292263

  9. Effects of host habitat quality on the viability of Cryptocotyle lingua (Trematoda: Digenea) cercariae.

    PubMed

    Cross, M A; Irwin, S W B; Fitzpatrick, S M

    2005-02-01

    In an earlier study it was shown that Cryptocotyle lingua cercariae, matured in Littorina littorea from a polluted marine lagoon, displayed slower horizontal swimming rates, and reduced longevity compared to cercariae released by periwinkles from a cleaner environment. This work investigated whether the pollution-induced reduction in swimming rates was due to an inefficient swimming action or the adoption of a less direct swimming path. In addition, cercariae from L. littorea that had been transferred from an 'unpolluted' to a 'polluted' site for 1 month provided information on the speed with which pollutants affect cercariae through their intermediate hosts. Results indicated that, in general, horizontal swimming rates were reduced due to slower swimming rather than disorientation and longer swimming pathways. Effects of host transplantation to a polluted site were clearly evident after 1 month. Evidence suggested that the pollutants accumulated by the cercariae via their first intermediate host affected the neuromusculature associated with swimming performance rather than sensory structures. Bearing in mind the reduced viability of C. lingua cercariae in polluted sites it is assumed that high prevalence of this digenean in gastropods (at such sites) must be due to their continual introduction by infected birds attracted to these habitats from other areas. PMID:15727069

  10. [Clonal variability in longevity of the cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

    PubMed

    2013-01-01

    The study was carried out on Himasthla elongata cercariae shed by infected Littorina littorea snails. The infected periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. The LT50 (the time during which 50% of cercariae perish in the experimental dish) were measured experimentally for cercariae Himasthla elongata belong to different clones. The investigated parameter demonstrated a high level of interclonal variability. Two groups of cercarial clones were identified: one of them was characterized by the high level of intraclonal variability in LT50 and the second, by the low one. It is assumed that the observed heterogeneity may be stipulated by different degrees of mitotic recombinations during formation of different cercarial clones. PMID:25508369

  11. [Clonal variability in longevity of the cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

    PubMed

    Levakin, I A; Losev, E A; Zavirskiĭ, Ia V; Galaktionov, K V

    2013-01-01

    The study was carried out on Himasthla elongata cercariae shed by infected Littorina littorea snails. The infected periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. The LT50 (the time during which 50% of cercariae perish in the experimental dish) were measured experimentally for cercariae Himasthla elongata belong to different clones. The investigated parameter demonstrated a high level of interclonal variability. Two groups of cercarial clones were identified: one of them was characterized by the high level of intraclonal variability in LT50 and the second, by the low one. It is assumed that the observed heterogeneity may be stipulated by different degrees of mitotic recombinations during formation of different cercarial clones. PMID:25464738

  12. Transmammary infection of free-ranging Florida panther neonates by Alaria marcianae (Trematoda: Diplostomatidae).

    PubMed

    Foster, Garry W; Kinsella, John M; Sheppard, Barbara J; Cunningham, Mark W

    2009-02-01

    Two freshly-dead female Florida panther (FP) neonates, Puma concolor couguar (=Puma concolor coryi), an 11-day-old and a 17-day-old, were collected in the Florida Panther National Wildlife Refuge (26 degrees 14'N, 81 degrees 36'W), Collier County, Florida. The 2 neonates were siblings and had presumably fed only on milk from the dam since birth. A 12-day-old female FP neonate was collected in the Big Cypress National Preserve (26 degrees 05'N, 81 degrees 15'W), Collier County, Florida and had also fed only on milk from the dam since birth. Milk was the only food item found in the gastrointestinal tract of these neonates. Mesocercariae and diplostomula of Alaria marcianae were collected from the lungs of the 3 neonates, indicating a transmammary route of infection. No mesocercariae, diplostomula, or mature A. marcianae were seen in the stomach or small intestine. The probable paratenic host for the A. marcianae infection in the adult Florida panther is the raccoon (Procyon lotor). PMID:18665664

  13. Cytochemical demonstration of cholinergic, serotoninergic and peptidergic nerve elements in Gorgoderina vitelliloba (Trematoda: Digenea).

    PubMed

    McKay, D M; Halton, D W; Johnston, C F; Fairweather, I; Shaw, C

    1991-02-01

    Standard enzyme cytochemical and indirect immunocytochemical techniques have been used in conjunction with light and confocal scanning laser microscopy (CSLM) to visualize cholinergic, serotoninergic and peptidergic nerve elements in whole-mount preparations of the amphibian urinary-bladder fluke, Gorgoderina vitelliloba. Cholinesterase (ChE) activity was localized in paired anterior ganglia, a connecting dorsal commissure and in the origins of the ventral nerve cords. Cholinergic ganglia were also evident in shelled embryos in the uterus. Serotonin-immunoreactivity (IR) was more extensive than ChE activity and was identified in both the central and peripheral nervous systems. Serotoninergic nerve fibres were associated with the somatic musculature and female reproductive ducts. Antisera to nine mammalian peptides and one invertebrate (FMRFamide) peptide have been used to investigate the peptidergic nervous system in the parasite. Immunoreactivity was obtained to five peptides, namely pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), substance P (SP) and FMRFamide. Peptidergic nerve fibres were found to be more abundant than demonstrable cholinergic or serotoninergic nerve fibres. NPY-IR was identified only in the main components of the central nervous system. However, PP- and PYY-IR occurred in the anterior ganglia, dorsal commissure, main nerve cords and in numerous small varicose fibres that ramified throughout the worm. Additionally, PP-immunoreactive nerve fibres were found to innervate the musculature of the female reproductive tracts. Six sites of IR were found in the acetabulum, using antisera directed towards the C-terminal end of PP and PYY, and these matched with the distribution of six non-ciliated rosette-like papillae observed by scanning electron microscopy. SP- and FMRFamide-IR were identified in the CNS, and FMRFamide-immunopositive nerve fibres were also evident in association with the gonopore cirrus region and with the terminal excretory pore. Results are discussed with respect to possible roles for each of the neurochemical types. PMID:2040570

  14. Phyllodistomum kanae sp. nov. (Trematoda: Gorgoderidae), a bladder fluke from the Ezo salamander Hynobius retardatus.

    PubMed

    Nakao, Minoru

    2015-10-01

    The Ezo salamander, Hynobius retardatus, is endemic only to Hokkaido, the northernmost island of Japan. Gravid flukes of the family Gorgoderidae were discovered from the urinary bladder of H. retardatus. The parasites were identified as a new species named Phyllodistomum kanae sp. nov. In the neighboring Honshu island another bladder fluke, Phyllodistomum patellare, has already been found from the Japanese newt. The new species clearly differs from P. patellare in having a spherical ovary and very weakly lobed testes. The discovery of species of Phyllodistomum from urodelan amphibians is very uncommon in Eurasia. A molecular phylogeny based on 28S ribosomal DNA suggests that sphaeriid bivalves may serve as the first intermediate host for the new species.

  15. Orientobilharzia Dutt & Srivastava, 1955 (Trematoda: Schistosomatidae), a junior synonym of Schistosoma Weinland, 1858.

    PubMed

    Aldhoun, Jitka A; Littlewood, D Timothy J

    2012-06-01

    In the last few decades, phylogenetic studies of the family Schistosomatidae based on molecular markers have revealed that members of the genus Orientobilharzia Dutt & Srivastava, 1955 belong within Schistosoma Weinland, 1858. In this study, the original descriptions and redescriptions of Orientobilharzia species and related revisions are reviewed, and it is confirmed that the morphological characters correspond with the results of the molecular studies. The two genera differ only in the number of testes; however, this character varies to a large extent within particular genera of the subfamily Schistosomatinae and cannot be used to justify the separation of Orientobilharzia from Schistosoma. Also, we have verified claims suggesting the synonymy of certain species of Orientobilharzia; the four valid species of this genus are transferred to Schistosoma and two new synonymies are formally presented. The following nomenclatural changes are made: Schistosoma Weinland, 1858 [syn. Orientobilharzia Dutt & Srivastava, 1955 (syn. nov.)]; Schistosoma bomfordi Montgomery, 1906 (comb. restit.); S. turkestanicum Skrjabin, 1913 (comb. restit.) [syns Ori. turkestanica var. tuberculata (Bhalerao, 1932) (syn. nov.) and Ori. cheni Hsü & Yang, 1957 (syn. nov.)]; S. dattai (Dutt & Srivastava, 1952) n. comb.; and S. harinasutai (Kruatrachue, Bhaibulaya & Harinasuta, 1965) n. comb. The generic diagnosis of Schistosoma is amended and a revised key to the subfamily Schistosomatinae Stiles & Hassall, 1898 is presented.

  16. MOLECULAR SYSTEMATICS OF THE AVIAN SCHISTOSOME GENUS TRICHOBILHARZIA (TREMATODA: SCHISTOSOMATIDAE) IN NORTH AMERICA

    PubMed Central

    Brant, Sara V.; Loker, Eric S.

    2010-01-01

    Trichobilharzia is a genus of thread-like schistosomes with a cosmopolitan distribution in birds. Species of Trichobilharzia achieve notoriety as major etiological agents of cercarial dermatitis, or swimmer’s itch. There are 40 species described in the literature, for which the majority lacks molecular sequence information. To better understand the phylogenetic relationships, diversity, species boundaries, host use, and geographic distribution of this genus, we surveyed 378 birds and over 10,000 snails from North America. The phylogenetic analysis was based on nuclear 18S, 28S rDNA, internal transcribed spacer region and mitochondrial cytochrome oxidase I sequence data. Specimens were recovered that could be related to 6 of the 14 described species of Trichobilharzia from North America (T. physellae, T. querquedulae, T. szidati, T. stagnicolae, T. franki, and T. brantae). An additional 5 lineages were found that could not be related directly to previously described species. Trichobilharzia brantae, transmitted by Gyraulus parvus, grouped outside the clade containing the recognized species of Trichobilharzia. A subgroup of the Trichobilharzia clade designated Clade Q was comprised of closely related species whose adults and eggs are similar, yet the European species use lymnaeids whereas the North American species use physids as snail hosts. This molecular phylogeny provides a useful framework to: 1) facilitate identification of worms, including those involved in dermatitis outbreaks; 2) test hypotheses about the evolution, diversification, host-parasite interactions and character evolution of Trichobilharzia; and 3) guide future taxonomic revision of Trichobilharzia. PMID:20049999

  17. Clinical, hematological, biochemical, and ultrasonographic aspects of Platynosomum sp. (Trematoda: Dicrocoeliidae) infection of captive Callithrix penicillata.

    PubMed

    Mattioli, Mariana Portugal; Batista, Juliana dos Santos; Ferrari, Marlon; Paludo, Giane Regina; Dias, Cecília Azevedo; Hoppe, Estevam G L; da Rocha, Gino Chaves; Monteiro, Rafael Veríssimo

    2016-04-01

    Trematodes from the genus Platynosomum have been found to infect Neotropical primates in captivity, but little is known about their pathogeny in such hosts. This study evaluated the physiological effects of natural infection by the liver-dwelling trematode Platynosomum sp. in ten males and ten females of Callithrix penicillata kept in captivity at the Primate Center of the University of Brasília. The marmosets were examined twice, 6 months apart. The following parameters were analyzed: complete blood count, bleeding time, serum total protein, albumin, and the liver enzymes AST and ALT, and both a stool analysis and a liver ultrasonic evaluation were performed. We were able to characterize a group of abnormalities associated with this trematode infection which were mainly derived from the hepatitis caused by it: coagulation disorders, abnormal red blood cells, hypoalbuminemia, and abnormal levels of liver-linked serum enzymes. Eosinophilia and thrombocytopenia were also commonly seen. All of the aforementioned abnormalities were in good accord with typical effects of trematodes on liver parenchyma. We suggest that this set of abnormalities is characteristic of the infection of C. penicillata with Platynosomum sp., and should be among the most prominent aspects that the veterinary surgeon considers when suspecting such an infection. We also suggest that these clinical signs and abnormalities will be similar in other liver-dwelling trematode-infected primate species. PMID:26922567

  18. [The oral sucker muscles of six representatives of the order Paramphistomatida (Plathelminthes, Trematoda)].

    PubMed

    Burdakova, E N; Yastrebova, I V; Yastrebov, M V

    2015-01-01

    The spatial arrangement of muscles in the oral suckers of six trematode species belonging to five families of the order Paramphistomatida is described. The functional load and adaptive significance of different muscle groups in the suckers themselves and the associated structures--preoral lip and muscular cap--are discussed. Complete section series in three projections have been examined; this demonstrates the presence of previously unnoticed structures, namely, semicircular and diagonal muscles, regulating shape of the oral cavity, and short longitudinal muscles, acting as a sphincter, as well as localization of the largest internal muscles on the sucker lateral sides. It has been shown that the presence of internal longitudinal muscles suggests that the organs in question are closer to the oral suckers of other trematodes rather than to their pharynxes.

  19. Malformations and mortality in the Asian Common Toad induced by exposure to pleurolophocercous cercariae (Trematoda: Cryptogonimidae).

    PubMed

    Jayawardena, Uthpala A; Tkach, Vasyl V; Navaratne, Ayanthi N; Amerasinghe, Priyanie H; Rajakaruna, Rupika S

    2013-06-01

    Malformations and increased mortality due to infection by the digenetic trematode, Riberioa ondatrae have been reported for many species of amphibians. Severe malformations have also been reported in the Common Hourglass Tree Frog, Polypedates cruciger induced by pleurolophocercous cercariae in Sri Lanka in addition to the changes in the behaviour, development and survival of the host. We exposed pre-limb bud stage tadpoles (Gosner stages 25-26) of the Asian Common Toad, Duttaphrynus melanostictus to the same pleurolophocercous type cercariae under laboratory conditions. Molecular and morphological identification showed that these cercariae belonged Acanthostomum burminis infecting freshwater snakes as definitive hosts. These cercariae induced malformations (27.8%) and reduced survival to metamorphosis (53.8%). The magnitude of the effects increased with the dose of cercariae. Types of malformations were mainly axial, such as scoliosis and kyphosis. Severe limb malformations such as extra or missing limbs as reported for amphibians exposed to R. ondatrae were not observed in the D. melanostictus. Same authors reported a higher percentage of malformations previously when P. cruciger was exposed to the cercariae A. burminis compared to D. melanostictus. However, tadpoles of D. melanostictus, which are smaller compared to those of P. cruciger, experienced higher mortality than P. cruciger tadpoles. Trematode induced malformations and mortality in amphibians are highly variable and depend on multiple factors such as host species differences such as resistance to infection and tolerance, life-history characteristics such as size at metamorphosis and length of the metamorphosis period, and other factors such as size of the amphibian at the time of trematode exposure.

  20. Phyllodistomum kanae sp. nov. (Trematoda: Gorgoderidae), a bladder fluke from the Ezo salamander Hynobius retardatus.

    PubMed

    Nakao, Minoru

    2015-10-01

    The Ezo salamander, Hynobius retardatus, is endemic only to Hokkaido, the northernmost island of Japan. Gravid flukes of the family Gorgoderidae were discovered from the urinary bladder of H. retardatus. The parasites were identified as a new species named Phyllodistomum kanae sp. nov. In the neighboring Honshu island another bladder fluke, Phyllodistomum patellare, has already been found from the Japanese newt. The new species clearly differs from P. patellare in having a spherical ovary and very weakly lobed testes. The discovery of species of Phyllodistomum from urodelan amphibians is very uncommon in Eurasia. A molecular phylogeny based on 28S ribosomal DNA suggests that sphaeriid bivalves may serve as the first intermediate host for the new species. PMID:25892565

  1. Studies on the life-cycle of Pleurogenoides ovatus Rao, 1977 (Trematoda: Pleurogenetinae).

    PubMed

    Janardanan, K P; Prasadan, P K

    1991-03-01

    The life-cycle of Pleurogenoides ovatus Rao, 1977, infecting the frogs, Rana tigrina and R. cyanophlyctis has been elucidated. All the life-cycle stages from egg to egg-producing adults were successfully established in the laboratory. The life-cycle took about 80 days for completion. Cercariae were found in the freshwater snail. Digoniostoma pulchella, collected from paddy fields at Chelembra, Malappuram district of Kerala, during the monsoon months. Cercariae are of the virgulate xiphidiocercous type. Metacercariae occurred in the connective tissues, hepatopancreas and musculature of the freshwater crab, Paratelphusa hydrodromous. The growth and development of the metacercariae in P. hydrodromous have been studied in detail. Frogs became infected when they fed on infected crabs. The prepatent period is 10 days.

  2. [A contribution to the biology of Dicrocoelium hospes Looss, 1907 (Trematoda, Dicrocoeliidae)].

    PubMed

    Lucius, R; Frank, W

    1978-06-01

    In this study, carried out in the Ivory Coast, Dicrocoelium hospes Looss, 1907 was shown for the first time to be a parasite of cattle and it was also detected in sheep by the presence of ova in the faeces. Up to 50% of the sheep in some flocks were found to shed eggs. The adults, sporocysts, "slime balls" and cercariae of D. hospes are described and compared to the respective stages of D. dendriticum. This comparison shows that essential morphological structures are very similar, especially the sensory papillae, which as so called chaetotaxis can serve to distinguish closely related species. The cercariae develop in terrestrial snails of the genus Limicolaria of which several species may play a role (L. flammea, L. felina, L. kambeul). The main habitat of D. hospes seems to be limited to savannah environment; only once could D. hospes be found in the borderzone between savannah and the rain forest. Infected Limicolaria-snails shed the cercariae in amorphous "slime balls" of liquid-jelly-like consistency and yellowish colour. Under the influence of sunlight these liquify; the cercariae, however, can survive for up to 26 h. According to our present knowledge the sunlight seems to be a factor which triggers the shedding of slime balls. This begins shortly after sunrise at a temperature of 22 degrees C and a relative humidity of 100%. Infected snails can shed cercariae on seven consecutive days. This kind of slime ball production might provide a clue to the 2nd intermediate host; efforts to identify this host have, however, so far been fruitless, despite the fact that almost 4000 arthropods, mainly ants, have been investigated. PMID:28656

  3. Malformations and mortality in the Asian Common Toad induced by exposure to pleurolophocercous cercariae (Trematoda: Cryptogonimidae).

    PubMed

    Jayawardena, Uthpala A; Tkach, Vasyl V; Navaratne, Ayanthi N; Amerasinghe, Priyanie H; Rajakaruna, Rupika S

    2013-06-01

    Malformations and increased mortality due to infection by the digenetic trematode, Riberioa ondatrae have been reported for many species of amphibians. Severe malformations have also been reported in the Common Hourglass Tree Frog, Polypedates cruciger induced by pleurolophocercous cercariae in Sri Lanka in addition to the changes in the behaviour, development and survival of the host. We exposed pre-limb bud stage tadpoles (Gosner stages 25-26) of the Asian Common Toad, Duttaphrynus melanostictus to the same pleurolophocercous type cercariae under laboratory conditions. Molecular and morphological identification showed that these cercariae belonged Acanthostomum burminis infecting freshwater snakes as definitive hosts. These cercariae induced malformations (27.8%) and reduced survival to metamorphosis (53.8%). The magnitude of the effects increased with the dose of cercariae. Types of malformations were mainly axial, such as scoliosis and kyphosis. Severe limb malformations such as extra or missing limbs as reported for amphibians exposed to R. ondatrae were not observed in the D. melanostictus. Same authors reported a higher percentage of malformations previously when P. cruciger was exposed to the cercariae A. burminis compared to D. melanostictus. However, tadpoles of D. melanostictus, which are smaller compared to those of P. cruciger, experienced higher mortality than P. cruciger tadpoles. Trematode induced malformations and mortality in amphibians are highly variable and depend on multiple factors such as host species differences such as resistance to infection and tolerance, life-history characteristics such as size at metamorphosis and length of the metamorphosis period, and other factors such as size of the amphibian at the time of trematode exposure. PMID:23353759

  4. Transmammary infection of free-ranging Florida panther neonates by Alaria marcianae (Trematoda: Diplostomatidae).

    PubMed

    Foster, Garry W; Kinsella, John M; Sheppard, Barbara J; Cunningham, Mark W

    2009-02-01

    Two freshly-dead female Florida panther (FP) neonates, Puma concolor couguar (=Puma concolor coryi), an 11-day-old and a 17-day-old, were collected in the Florida Panther National Wildlife Refuge (26 degrees 14'N, 81 degrees 36'W), Collier County, Florida. The 2 neonates were siblings and had presumably fed only on milk from the dam since birth. A 12-day-old female FP neonate was collected in the Big Cypress National Preserve (26 degrees 05'N, 81 degrees 15'W), Collier County, Florida and had also fed only on milk from the dam since birth. Milk was the only food item found in the gastrointestinal tract of these neonates. Mesocercariae and diplostomula of Alaria marcianae were collected from the lungs of the 3 neonates, indicating a transmammary route of infection. No mesocercariae, diplostomula, or mature A. marcianae were seen in the stomach or small intestine. The probable paratenic host for the A. marcianae infection in the adult Florida panther is the raccoon (Procyon lotor).

  5. Phyllodistomum folium (Trematoda: Gorgoderidae) infecting zebra mussels (Dreissena polymorpha) in the Ebro River, Spain.

    PubMed

    Peribáñez, Miguel A; Elrío, María L; Gracia, María J; Fernández de Luco, Daniel; Castillo, Juan A; Lucientes, Javier; Cia, Imanol

    2006-06-01

    Zebra mussels (Dreissena polymorpha) were first found in the Ebro River (Spain) in Ribaroja reservoir, in the summer of 2001. This paper reports a study to detect parasites in this bivalve species. From September 2003 to August 2004, a total of 1380 zebra mussels were collected and dissected or sectioned in paraffin and haematoxylin and eosin staining. We observed the presence of Phyllodistomum folium (Olfers, 1816) in two hosts (prevalence 0.14%). Sporocysts containing metacercariae were located within the gill lamellae. One of the mussels was collected in January and the other one in July. In both cases the shell length was >2 cm. P. folium had not been previously reported in Spain and D. polymorpha is its only known intermediate host. It represents a new invasive species in this river basin, presumably introduced together with the zebra mussels.

  6. Chromosome analysis of Phyllodistomum folium (Trematoda, Gorgoderidae) infecting three European populations of zebra mussels.

    PubMed

    Petkeviciūte, R; Staneviciūte, G; Molloy, D P

    2003-08-01

    The mitotic chromosome sets of the larval stages of Phyllodistomum folium infecting three European populations of Dreissena polymorpha were studied using conventional Giemsa staining and karyometrical analysis. The karyotype, described herein for the first time, consisted of nine chromosome pairs (2 n=18). The chromosomes were comparatively small and measured from 1.19 micro m to 5.22 micro m. The first and second longest chromosome pairs were, respectively, approximately 20% and 18% of the mean total chromosome set length and were clearly differentiated from the other seven pairs, which gradually decreased in size. Most chromosomes were biarmed with medially or submedially located centromeres; only pairs 1 and 4 contained subterminally located centromeres. No significant differences in the relative length values of corresponding chromosomes were observed between the three populations studied. The main interpopulation differences were detected in the centromeric index values of some corresponding chromosome pairs. These data, when analysed in conjunction with those of other Gorgoderidae, indicate that the karyotype of P. folium and other species in this family do not show any clear affinities with "typical" plagiorchiate chromosome sets.

  7. Comparison of the hemoglobins of the platyhelminths Gastrothylax crumenifer and Paramphistomum epiclitum (Trematoda: Paramphistomatidae).

    PubMed

    Haque, M; Rashid, K A; Stern, M S; Sharma, P K; Siddiqi, A H; Vinogradov, S N; Walz, D A

    1992-04-01

    1. Gastrothylax crumenifer and Paramphistomum epiclitum parasitize the water buffalo Bubalus bubalis. 2. Gastrothylas hemoglobin consisted of two fractions of ca 30,000 and ca 18,000 by gel filtration. SDS-electrophoresis showed both to be single, ca 15,000 chains. 3. Paramphistomum hemoglobin was ca 16,000 by both gel filtration and SDS-electrophoresis. 4. Reversed-phase chromatography of carboxymethylated trematode and buffalo globins gave single peaks and two peaks, respectively. Although Paramphistomum hemoglobin provided and N-terminal sequence, Gastrothylax hemoglobin did not, suggesting blocked N-terminals. The buffalo sequences were found to be identical to the sequences of the alpha and beta chains of bovine hemoglobin. 5. Although Paramphistomum hemoglobin consists of only one chain, Gastrothylax hemoglobin consists either of one chain which aggregates to a dimer or of two different chains, only one of which aggregates to a dimer.

  8. Characterization of the glutamate dehydrogenase activity of Gigantocotyle explanatum and Gastrothylax crumenifer (Trematoda: Digenea).

    PubMed

    Abidi, S M A; Khan, P; Saifullah, M K

    2009-12-01

    Glutamate dehydrogenase (GLDH) (EC 1.4.1.3) is a ubiquitous enzyme, which is present at the protein and carbohydrate metabolism crossroads. The enzyme activity was investigated in biliary and rumen amphistomes, Gigantocotyle explanatum and Gastrothylax crumenifer, respectively, infecting the Indian water buffalo Bubalus bubalis. The enzyme activity was consistently higher in G. explanatum as compared to G. crumenifer, where NAD(H) was utilized as coenzyme and the pH optima was recorded at 8. The K(m) and V(max) values for α-ketoglutarate were 2.1 mM and 9.09 units in G. explanatum, whereas 3.03 mM and 1.90 units in G. crumenifer, respectively. Among the allosteric modulator nucleotides, AMP, ADP, ATP, GMP, CMP and UMP, only AMP enhanced GLDH activity in G. crumenifer while ADP was stimulatory in G. explanatum. The amino acid leucine stimulated the GLDH activity in both the amphistomes while alanine was stimulatory only in G. crumenifer. Pronounced interspecific differences in response to different metabolic inhibitors like diethyldithiocarbamate, semicarbazide hydrochloride and mercurial ions were also observed. The osmotic stress alters the enzyme activity, particularly in hypertonic saline the GLDH activity increased significantly (p < 0.01) in G. explanatum, while insignificant effects were observed in rumen dwelling G. crumenifer. Histoenzymology revealed region/tissue specific distribution of GLDH with prominent staining in tissues like vitellaria, lymph system and tegument/subtegument, thus showing specific distribution of GLDH indicating differential metabolic state. Such intergeneric differences in GLDH activity could also be a consequence of occupying different microenvironments within the same host.

  9. Novel Infection Site and Ecology of Cryptic Didymocystis sp. (Trematoda) in the Fish Scomberomorus maculatus.

    PubMed

    Schrandt, Meagan N; Andres, Michael J; Powers, Sean P; Overstreet, Robin M

    2016-06-01

    An undescribed, cryptic species of Didymocystis, as determined from sequences of 2 ribosomal genes and superficially similar to Didymocystis scomberomori ( MacCallum and MacCallum, 1916 ), infected the skin of the Spanish mackerel, Scomberomorus maculatus , in the north-central Gulf of Mexico (GOM). An analysis of 558 fish from 2011 to 2013 from Louisiana, Mississippi, Alabama, and the Florida panhandle showed the prevalence of the trematode varied both spatially and temporally but not with sex of the fish host. Month, year, and geographic location were identified by a negative binomial generalized linear model as indicators of the abundance and intensity of infection. Prevalence, abundance, and intensity of infection were greatest in spring and fall months off the Florida panhandle. Furthermore, the abundance and intensity of infection correlated negatively with fork length, weight, and gonad weight of mature fish but positively with longitude. Therefore, smaller adult fish tended to be more infected than larger adults, and prevalence and intensity increased from west to east (Louisiana to Florida). Spatial and temporal trends seemed to result from physical factors (e.g., water temperature, salinity, bottom type), but they also coincided with the annual migration of S. maculatus as fish moved northward along the GOM coastline from the southern tip of Florida in the spring months and returned in the fall, being present in the north-central GOM from late spring through fall. This pattern suggests the possibility that acquisition of infections occurred from a molluscan host in waters off the Florida panhandle.

  10. Further studies on in vitro pairing of Echinostoma revolutum (Trematoda) adults.

    PubMed

    Fried, B; Pallone, A

    1984-01-01

    Various factors that influence in vitro pairing of Echinostoma revolutum adults were studied. More worms paired in Locke's, Ringer's, and 0.85% NaCl solutions than in the defined medium NCTC 135. Also, more pairing occurred at 39 or 43 degrees C than at 35 degrees C. Echinostomes placed 1, 3, or 5 cm apart in 5.5 cm diameter Petri dishes as well as those placed 2, 4, or 6 cm apart in 8.5 cm diameter Petri dishes paired; however, worms placed 8 cm apart in the latter dishes did not. Individuals maintained in Locke's solution at 20 or 39 degrees C for 1 h after having been removed from the chicks paired less frequently than those used for experiments immediately after their removal from chicks. Worms showed more pairing in fresh Locke's solution than in this solution preconditioned by excretory-secretory products of E. revolutum. Scanning electron microscopy revealed no differences between the tegument of fresh worms and of those maintained under experimental conditions for 2 h at 39 degrees C. PMID:6741219

  11. Egg laying in vitro of Echinostoma caproni (Trematoda) in nutritive and nonnutritive media.

    PubMed

    Reddy, A; Fried, B

    1996-01-01

    Egg laying in vitro was studied in Echinostoma caproni adults placed in 10 ml of nutritive or nonnutritive media for 48 h in petri-dish cultures maintained at 37 degrees C in an atmosphere containing 7.6% CO2. Maximal egg laying occurred within 24 h in the defined medium RPMI 1640. Egg laying was significantly greater in this medium than in McCoy's or Locke's solution. Eggs released into the RPMI medium were capable of producing miracidia that were infective to Biomphalaria glabrata snails. Fried and Huffman (1996) referred to a technique used to obtain eggs of Echinostoma caproni in the defined medium RPMI 1640, but details of the study were not given. No information is available on egg laying of echinostomes in vitro. Such information could contribute to a better understanding of egg release in digeneans and would also be helpful in the acquisition of eggs for biology and chemistry studies. Current techniques used to obtain echinostome eggs involve worm homogenization, teasing of eggs from the worms' uteri, or recovery of eggs from feces (see Idris and Fried 1996 for details). The purpose of this communication is to report on an efficient procedure for the acquisition of eggs of E. caproni after the placement of adult worms in the defined medium RPMI 1640. E. caproni adults were grown in ICR mice for either 17 (young worms) or 112 days (old worms) as described previously (Ursone and Fried 1995a). Worms were removed from the small intestines and rinsed rapidly in three changes of sterile Locke's solution containing penicillin (200 IU/ml) and streptomycin (200 micrograms/ml; Fried and Contos 1973). Worms were placed in culture media within 30 min of their removal from hosts. Nutritive media consisted of RPMI 1640 and McCoy's medium (Sigma, St. Louis, Mo.). Non-nutritive media consisted of Locke's or Locke's 1:1 (Ursone and Fried 1995b). All media contained antibiotics as described above. PMID:8738289

  12. Two new species of Phyllodistomum Braun, 1899 (Trematoda: Gorgoderidae Looss, 1899) from Great Barrier Reef fishes.

    PubMed

    Ho, Hei Wa; Bray, Rodney A; Cutmore, Scott C; Ward, Selina; Cribb, Thomas H

    2014-03-19

    Two new species of Phyllodistomum Braun, 1899 are described from the urinary bladder of fishes of the Great Barrier Reef. Phyllodistomum hoggettae n. sp. is described from Plectropomus leopardus (leopard coralgrouper) (Serranidae) and P. vaili n. sp. is described from Mulloidichthys vanicolensis (yellowfin goatfish) and M. flavolineatus (yellowstripe goatfish) (Mullidae). These species are compared with 26 previously described marine Phyllodistomum species and found to be distinct in combinations of body shape, sucker ratio and shape of the gonads. Preliminary molecular data also demonstrate that they are distinct from each other and for those other species for which data are available.

  13. Comparative efficacy of anthelmintics and their effects on hemato-biochemical changes in fasciolosis of goats of South Gujarat

    PubMed Central

    Shrimali, R. G.; Patel, M. D.; Patel, R. M.

    2016-01-01

    Aim: Fasciolosis is a parasitic disease caused by Fasciola spp. of the family Fasciolidae (trematodes) characterized by bottle jaw, anemia, progressive debility, and potbelly condition. There are many aspects of fasciolosis remaining unknown thus hemato-biochemical alterations in closantel, triclabendazole + ivermectin, and oxyclozanide + levamisole treated goats were studied. Materials and Methods: A total of 40 naturally fasciolosis infected goats having egg per gram more than 100 were randomly divided into four groups. Goats of Group I-III were treated with three different anthelmintics, whereas, goats of Group-IV were kept as control or untreated. Whole blood, serum, and fecal samples were collected on 0, 7th, and 30th day of treatment. Results: During the study, values of hemoglobin, total erythrocyte count, pack cell volume, and total protein were significantly elevated to their normal levels in anthelmintics treated groups. Whereas, values of total leukocyte count, aspartate transaminase (AST), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) were significantly reduced to their normal level in anthelmintics treated groups. The efficacy of closantel (T1), triclabendazole + ivermectin (T2), and oxyclozanide + levamisole (T3) was 99.63%, 100%, and 94.74% and 100%, 100%, and 97.38% on 7th and 30th day of treatment, respectively. Conclusions: Fasciolosis in goats can be diagnosed on the basis of fecal sample examination, but alterations in important biomarkers such as AST, GGT, and LDH are also helpful for early diagnosis. The use of newer anthelmintic either alone or in combination showed a higher therapeutic response in fasciolosis of goats. PMID:27284231

  14. Phylogeography and genetic divergence of some lymnaeid snails, intermediate hosts of human and animal fascioliasis with special reference to lymnaeids from the Bolivian Altiplano.

    PubMed

    Jabbour-Zahab, R; Pointier, J P; Jourdane, J; Jarne, P; Oviedo, J A; Bargues, M D; Mas-Coma, S; Anglés, R; Perera, G; Balzan, C; Khallayoune, K; Renaud, F

    1997-04-15

    A population genetic study using starch gel electrophoresis was performed on populations of several species of lymnaeid snails acting as intermediate hosts for Fasciola hepatica (Trematoda, Plathyhelminth). Lymnaea viatrix was collected in 16 sites from the Bolivian Northern Altiplano. L. cubensis were obtained in one site from Venezuela, one site from Guadeloupe, three sites from Cuba and one site from the Dominican Republic. L. truncatula were collected in one site from France, one from Portugal and one from Morocco. Multilocus enzyme electrophoresis (MEE) were determined for 282 snails at 18 loci. A complete monomorphism was encountered at each geographic site. However, among these 18 loci, 13 are polymorphic and low and high levels of genetic divergence were observed between samples. Two genotypic groups can be differentiated by their multilocus genotypes. The western genotypic group associates together samples from Venezuela, Guadeloupe, Cuba and Dominican Republic (L. cubensis) while samples from France, Portugal and Morocco (L. truncatula) belong to the eastern genotypic group. Surprisingly, the Northern Bolivian Altiplano populations (L. viatrix) do not present any genetic divergence with the Portuguese sample. Therefore, the Bolivian snails belong entirely to the eastern genetic group. Within each group slight genetic divergences were observed. These results strongly support the European origin of the lymnaeid snails from the Northern Bolivian Altiplano.

  15. Larval stages of the bluefin tuna blood fluke Cardicola opisthorchis (Trematoda: Aporocotylidae) found from Terebella sp. (Polychaeta: Terebellidae).

    PubMed

    Sugihara, Yukitaka; Yamada, Toshiyuki; Tamaki, Akio; Yamanishi, Ryohei; Kanai, Kinya

    2014-04-01

    We found aporocotylid larval stages (sporocysts and cercariae) from five individuals of terebellid polychaete Terebella sp., which were collected from seabed substrate and ropes and floats attached to tuna cages in a tuna farm on the coast of Tsushima Island, Nagasaki, Japan. Nucleotide sequences of the regions of internal transcribed spacer 2 ribosomal DNA and 28S ribosomal DNA from these larval stages were 100% identical to those of Cardicola opisthorchis registered in GenBank. C. opisthorchis is a pathogen causing blood fluke infection of Pacific bluefin tuna Thunnus orientalis, which is considered to have a significant impact on the Japanese Pacific bluefin tuna aquaculture industry. This is the first description of the intermediate host of C. opisthorchis. This indicates that the life cycle of C. opisthorchis is completed within tuna farms in this area.

  16. The first comprehensive study of praziquantel effects in vivo and in vitro on European liver fluke Opisthorchis felineus (Trematoda).

    PubMed

    Pakharukova, Mariya Y; Shilov, Alexander G; Pirozhkova, Darya S; Katokhin, Alexey V; Mordvinov, Viatcheslav A

    2015-07-01

    The European liver fluke Opisthorchis felineus (Rivolta, 1884) is an epidemiologically important parasite infecting mammals, including humans. Opisthorchis felineus is widespread in Russia, Kazakhstan and Eastern European countries. Praziquantel (PZQ) is the drug of choice for the treatment of opisthorchiasis, but the effects of this drug on O. felineus are poorly studied. The aims of this work were (i) to perform a study of PZQ effects in vitro, (ii) to identify morphological markers of PZQ action on O. felineus, (iii) to analyse damage to the worm surface and (iv) to assess the efficacy of PZQ in vivo in a hamster model. Light microscopy, optical sectioning and fluorescence microscopy were used to study morphological changes. In vivo, PZQ at a dose of 400mg/kg reduced the rate of infection in experimental acute and chronic opisthorchiasis in hamsters by 70% and 79%, respectively. In vitro, the drug caused destruction and vacuolisation of the tegument of O. felineus, contractions of the worm musculature, paralysis, and irreversible changes in morphology (IC50=0.14μg/mL). Differences in susceptibility to the drug between adult and newly excysted metacercariae were also observed. Qualitative effects of PZQ in vivo and in vitro were similar to the drug's effects on other trematodes, including epidemiologically important liver flukes. Nevertheless, high heterogeneity of O. felineus specimens in terms of susceptibility to the drug was observed. In addition, we describe for the first time the high rate of recovery of O. felineus following the destructive action of PZQ. PMID:25862308

  17. Peracreadium akenovae sp. nov. (Trematoda: Opecoelidae) parasitising the highfin moray eel Gymnothorax pseudothyrsoideus (Anguilliformes: Muraenidae) from Moreton Bay, Australia.

    PubMed

    Cribb, Thomas H; Bray, Rodney A; Cutmore, Scott C

    2013-09-01

    A new opecoelid trematode, Peracreadium akenovae sp. nov., is described from the highfin moray eel Gymnothorax pseudothyrsoideus (Bleeker) (Anguilliformes; Muraenidae), collected from Moreton Bay off southeast Queensland, Australia. The new species is distinctive in its body shape, transversely elongate irregular testes, vitelline follicles interrupted at the level of the ventral sucker, and diverticulate excretory vesicle. The Muraenidae is a new host family for Peracreadium Nicoll, 1909. Peracreadium is the seventh opecoelid genus reported from temperate eastern Australian marine fishes and this is its first report from Australian waters.

  18. Integrative taxonomy of central European parasitic flatworms of the family Prosthogonimidae Lühe, 1909 (Trematoda: Plagiorchiida).

    PubMed

    Heneberg, Petr; Sitko, Jiljí; Bizos, Jiří

    2015-10-01

    Species of the family Prosthogonimidae are considered the most pathogenic poultry trematodes worldwide, affecting particularly low intensity farming in rural areas. Adults of Prosthogonimus occur mainly in the bursa of Fabricius, oviduct and cloaca of ducks, geese, fowl and other birds feeding at least occasionally on dragonflies or damselflies (Odonata). We analyzed the central European species of the Prosthogonimidae, namely Prosthogonimus cuneatus, Prosthogonimus ovatus, Prosthogonimus pellucidus and Prosthogonimus rarus. We sequenced three nuclear (ITS2) and mitochondrial (CO1, ND1) DNA loci of four species isolated from Anas clypeata, Anas strepera, Anas platyrhynchos, Aythya ferina, Passer domesticus and Turdus merula. Intra- and inter-specific sequence variability revealed that all four species represent distinct well-defined entities. Our data, combined with previously published studies, suggest the return of the name Prosthogonimus rarus Braun, 1901 for Schistogonimus rarus (Braun, 1901). The genus name Schistogonimus Lühe, 1909 is considered a junior synonym of Prosthogonimus Lühe, 1899. We identified the existence of two clades, one represented by P. cuneatus and P. pellucidus, and another one formed by P. ovatus and P. rarus. We also provide comparative measurements of these four central European prosthogonimids, and address their tissue specificity, host-specific prevalence (based on the extensive bird cohort examined in years 1962-2014), and for some bird hosts we address also differences in the prevalence of Prosthogonimus spp. in natural and near-natural wetlands in comparison with fishponds utilized for intense carp production. We provide an updated key to European Prosthogonimus spp. based on their morphological characters. PMID:25724855

  19. Description of a new lung fluke species, Paragonimus vietnamensis sp. nov. (Trematoda, Paragonimidae), found in northern Vietnam.

    PubMed

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

    2007-11-01

    Based on morphology of metacercariae and adult worms together with molecular data from our previous study, we describe herein a new lung fluke species named Paragonimus vietnamensis sp. nov. Metacercariae of this new species is round and extremely large in size (nearly 800 mum in diameter) having a fragile outer cyst of variable thickness and a thin fragile inner cyst. There are little or no space between metacercaria and the inner cyst wall. These and other morphological features of metacercariae of P. vietnamensis sp. nov. are not completely identical with but have some similarities with those of P. microrchis, P. proliferus, or P. menglaensis. On the other hand, adult worms obtained by experimental infection of these large metacercariae are oval in shape, having a ventral sucker slightly larger than the oral one, and having singly arranged relatively short cuticular spines. These morphological features of adult worms are partly similar to but not identical with those of P. microrchis, P. skrjabini, P. yunnanenis, P. xiangshanensis, or P. harinasutai. Taken these morphological data and our previous molecular phylogenetic analyses of ITS2 and CO1 sequences of metacercariae and adult worms together, P. vietnamensis sp. nov. is different from any other known Paragonimus spp.

  20. Sympatric and allopatric experimental infections of the planorbid snail Gyraulus chinensis with miracidia of Euparyphium albuferensis (Trematoda: Echinostomatidae).

    PubMed

    Muñoz-Antoli, C; Marín, A; Trelis, M; Toledo, R; Esteban, J-G

    2010-12-01

    An experimental infection with echinostomatid miracidia in sympatric or 'local' vs. allopatric or 'away' snail combinations, as a model to examine parasite compatibility, was carried out. We employed Euparyphium albuferensis miracidia to infect Gyraulus chinensis snails, from three different natural parks: Albufera (Valencia, Spain); the Ebro Delta (Tarragona, Spain) and Coto de Doñana (Huelva, Spain). Insignificant differences between the three snail strains were noted for the infection rate and the rhythm of daily cercarial production. However, a significantly higher total cercarial production per snail, patent period and life span were observed in local snails. The different infection characteristics in the three G. chinensis strains considered reveal that E. albuferensis miracidia demonstrate local adaptation.

  1. [Contribution to the study of Microphallidae Travassos, 1920 (Trematoda). XXXIV. Redescription of Levinseniella howensis Johnston, 1917 (author's transl)].

    PubMed

    Pearson, J C; Deblock, S

    1979-01-01

    According to the type of L. howensis the species belongs to the subgenus Levinseniella instead of the subgenus Monarrhenos as believed before. The genital atrium has two male pockets and one female pouch.

  2. Muscle architecture during the course of development of Diplostomum pseudospathaceum Niewiadomska, 1984 (Trematoda, Diplostomidae) from cercariae to metacercariae.

    PubMed

    Petrov, A; Podvyaznaya, I

    2016-05-01

    Recent confocal microscopy studies have greatly expanded our knowledge of muscle systems in cercariae and adult digeneans, but the gross anatomy and development of metacercarial musculature remain relatively little known. To further our understanding of metacercarial development, this study used phalloidin staining and confocal microscopy to examine changes in muscle architecture over the course of development from cercariae to infective metacercariae in Diplostomum pseudospathaceum Niewiadomska, 1984. The paper describes muscle development in the body wall, anterior organ (oral sucker), acetabulum, pharynx and midgut and in the musculo-glandular organs that first appear in metacercariae (lappets and holdfast). The muscle architecture of the cercarial tail is also described. The results of the study support previously reported observations that diplostomid musculature undergoes substantial transformation during metacercarial development. The most profound changes, involving extensive remodelling and replacement of cercarial muscles, were seen in the body-wall musculature and in the anterior organ as it developed into the oral sucker. Muscle systems of other cercarial organs showed more gradual changes. The adaptive importance of developmental changes in musculature is discussed.

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

    PubMed Central

    AZIZI, Hekmat; FARAHNAK, Ali; MOBEDI, Iraj; MOLAEI RAD, MohamadBagher

    2015-01-01

    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. PMID:25904952

  4. The cercaria of Galactosomum bearupi Pearson, 1973 (Trematoda: Heterophyidae) at Heron and Masthead Islands, Great Barrier Reef, Australia.

    PubMed

    Beuret, J; Scott, D A; Pearson, J C

    2000-05-01

    Magnacercous cercariae, all morphologically identical but with different coloured tails, were found naturally infecting the intertidal prosobranch gastropod Clypeomorus batillariaeformis Habe & Kosuge (Cerithiidae) at Heron and Masthead Islands. Several species of coral-dwelling fishes were exposed to magnacercous cercariae. Active ingestion by the fish, followed by the complete development of the metacercaria in the optic lobes, occurred in Pomacentrus molluccensis Bleeker, but not in Dascyllus aruanus (L.), where development stopped short after encystment. All cercariae turned out to belong to the same species, Galactosomum bearupi Pearson, 1973. Natural infections of G. bearupi metacercariae were found in eight species of fish at Heron Island: Pomacentrus molluccensis, P. wardi Whitley, P. bankanensis Bleeker, P. flavicauda Whitley, Stegastes cf. fasciolatus (Ogilby), Sillago maculata Quoy & Gaimard, S. cf. ciliata Cuvier and Crenimugil crenilabris (Forsskål). This represents the fourth account of the cercaria of a species of Galactosomum.

  5. First documentation and molecular confirmation of three trematode species (Platyhelminthes: Trematoda) infecting the polychaete Marenzelleria viridis (Annelida: Spionidae).

    PubMed

    Phelan, Krystin; Blakeslee, April M H; Krause, Maureen; Williams, Jason D

    2016-01-01

    Polychaete worms are hosts to a wide range of marine parasites; yet, studies on trematodes using these ecologically important species as intermediate hosts are lacking. During examination of the spionid polychaete Marenzelleria viridis collected on the north shore of Long Island, New York, putative trematode cysts were discovered in the body cavity of these polychaetes. In order to verify these cysts as metacercariae of trematodes, specimens of the eastern mudsnail Ilyanassa obsoleta (a very common first intermediate host of trematodes in the region) were collected for molecular comparison. DNA barcoding using cytochrome C oxidase I regions confirmed the presence of three species of trematodes (Himasthla quissetensis, Lepocreadium setiferoides, and Zoogonus lasius) in both M. viridis and I. obsoleta hosts. Brown bodies were also recovered from polychaetes, and molecular testing confirmed the presence of L. setiferoides and Z. lasius, indicating an immune response of the polychaete leading to encapsulation of the cysts. From the 125 specimens of M. viridis collected in 2014, 95 (76.8 %) were infected with trematodes; of these 95 infected polychaetes, 86 (90.5 %) contained brown bodies. This is the first confirmation that trematodes use M. viridis as a second intermediate host and that this intermediate host demonstrates a clear immune response to metacercarial infection. Future research should explore the role of these polychaetes in trematode life cycles, the effectiveness of the immune response, and transmission pathways to vertebrate definitive hosts.

  6. Molecular characterization of Stictodora tridactyla (Trematoda: Heterophyidae) from Kuwait Bay using rDNA ITS and mtCO1.

    PubMed

    Al-Kandari, Wafa Y; Alnaqeeb, Majed A; Isaac, Asha M; Al-Bustan, Suzanne A

    2015-11-01

    Stictodora tridactyla is an intestinal fluke in the family Heterophyidae that parasitizes shorebirds and mammals, including humans. Its metacercarial cyst stage was reported in the Arabian killifish, Aphanius dispar, at Kuwait Bay. In the present study, Cerithidea cingulata was found to serve as the first intermediate host of S. tridactyla. In order to establish the snail-fish link in the life cycle of S. tridactyla, complete sequences of ribosomal DNA internal transcribed spacer region 1 and 2 (rDNA ITS1 and ITS2) and partial sequence of cytochrome oxidase subunit 1 were obtained for metacercarial cysts isolated from the fish A. dispar and rediae isolated from the snail C. cingulata. Sequence alignment demonstrated that these larval stages belong to the same heterophyid species, S. tridactyla. Phylogenetic analysis based on rDNA ITS1, ITS2, and mtCO1 confirmed the position of S. tridactyla within the Heterophyidae and found it to cluster with Haplorchis spp. The present study represents the first molecular study correlating the larval stages of S. tridactyla using rDNA ITS1, ITS2, and mtCO1 and examining the phylogenetic relationships of S. tridactyla with different heterophyid species. PMID:26268569

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

    PubMed

    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. PMID:22375817

  8. Morphological variation in Plagiorchis noblei Park 1936 (Trematoda: Plagiorchiidae) from Tyrannus tyrannus and T. verticalis (Aves: Tyrannidae).

    PubMed

    MacKenzie, D I; McKenzie, C E

    1980-02-01

    Specimens of Plagiorchis noblei from Tyrannus tyrannus and T. verticalis, were analyzed for morphological differences using multivariate techniques. Hotelling's T2-test showed a highly significant difference (P less than 0.001) between the two parasite populations. A stepwise discriminant analysis selected pharynx and testis as the characters that best separated the two populations, and correctly classified 69% of the specimens as to their host species. Plagiorchis gonzalchavezi was determined to be a junior subjective synonym of P. noblei. PMID:7189212

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

    PubMed

    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. PMID:12053986

  10. [Demonstration of the existence of a marine cycle in the strigeides: Cardiocephalus longicollis Szidat, 1928 (Trematoda: Strigeidae)].

    PubMed

    Prevot, G; Bartoli, P

    1980-01-01

    During their investigations on parasitism of fishes and birds in the lagoon of Brusc (Var), the authors have discovered the life cycle of Cardiocephalus longicollis. Cercariae of pharyngeate furcocercous type develop in sporocysts in the digestive gland of the marine prosobranch Nassa corniculum. The cercaria is identified as Cercaria nassae (Dolgikh, 1965) from Nassa reticulata in Black sea; it closely resembles Cercaria nassa (Martin, 1945) from Ilyanassa obsolseta (syn. Nassa obsoleta) at Woods Hole (U.S.A.). Several intermediate hosts have been related; they are the marine fishes Diplodus annularis, D. vulgaris, Boops salpa, Pagellus mormyrus (Sparidae) and Belone belone (Scombresocidae). Metacercariae of tetracotyle type were found in the cavity of optical lobes of the brain. Experimental contamination of fishes: Potamoschistus microps, Blennius sp., Mugil cephalus and Syngnathus abaster has been negative; that of Diplodus annularis and D. sargus, positive. It allowed to follow the development from cercaria to tetracotyle. In Provence natural definitive hosts are Larus argentatus and L. ribidundus; contaminations of these birds with metacercariae experimentally raised in Diplodus annularis and D. vulgaris were successfull. Some observations are pointed out especially the development of the parasite in the definitive host, the organisation of tribocytic organ and the geographical distribution of the parasite.

  11. Biomphalaria straminea (Mollusca: Planorbidae) as an intermediate host of Drepanocephalus spp. (Trematoda: Echinostomatidae) in Brazil: a morphological and molecular study.

    PubMed

    Pinto, Hudson A; Griffin, Matt J; Quiniou, Sylvie M; Ware, Cynthia; Melo, Alan L

    2016-01-01

    Species of trematodes belonging to the genus Drepanocephalus are intestinal parasites of piscivorous birds, primarily cormorants (Phalachrocorax spp.), and are widely reported in the Americas. During a 4-year malacological study conducted on an urban lake in Brazil, 27-collar-spined echinostome cercariae were found in 1665/15,459 (10.7 %) specimens of Biomphalaria straminea collected. The cercariae were identified as Drepanocephalus spp. by sequencing the 18S (SSU) rDNA, ITS1/5.8S rDNA/ITS2 (ITS), 28S (LSU) rDNA region, cytochrome oxidase subunit 1 (CO1), and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) markers. In experimental life cycle studies, metacercariae developed in laboratory-reared guppies (Poecilia reticulata); however, attempts to infect birds and rodents were unsuccessful. Two closely related morphotypes of cercariae were characterized. One species, identified by molecular markers as a genetic variant of Drepanocephalus auritus (99.9 % similarity at SSU, ITS, LSU; 97.2 % at CO1; 95.8 % at ND1), differs slightly from an archived North American isolate of this species also sequenced as part of this study. A second species, putatively identified as Drepanocephalus sp., has smaller cercariae and demonstrates significant differences from D. auritus at the CO1 (11.0 %) and ND1 (13.6 %) markers. Aspects related to the morphological taxonomic identification of 27-collar-spined echinostome metacercariae are briefly discussed. This is the first report of the involvement of molluscs of the genus Biomphalaria in the transmission of Drepanocephalus and the first report of D. auritus in South America.

  12. Experimental and molecular study of cercariae of Clinostomum sp. (Trematoda: Clinostomidae) from Biomphalaria spp. (Mollusca: Planorbidae) in Brazil.

    PubMed

    Pinto, H A; Caffara, M; Fioravanti, M L; Melo, A L

    2015-02-01

    Despite the large number of reports of species of Clinostomum from vertebrate hosts in South America, studies evaluating the molluscan transmitters of these parasites are scarce. In the present study, clinostomatoid cercariae shed from 0.02% (4/17,485) specimens of Biomphalaria spp., collected at the Pampulha reservoir, Belo Horizonte, Minas Gerais, Brazil, were used for experimental infection of Poecilia reticulata . Samples of cercariae from molluscs and metacercariae experimentally obtained from fish were subjected to morphological and molecular analyses and compared with species of Clinostomum reported in the Americas. The cercariae and metacercariae, here identified as Clinostomum sp., present general morphology similar to that reported for Clinostomum marginatum , however, from molecular point of view, differ significantly from North American C. marginatum and other species of Clinostomum reported in South America. These results suggest that the diversity of Clinostomum found in Brazil may be underestimated. Additional studies aimed at molecular characterization of South American species of Clinostomum, including the finding of specimens with sequences similar to that reported for C. marginatum in North America are required.

  13. Biomphalaria straminea (Mollusca: Planorbidae) as an intermediate host of Drepanocephalus spp. (Trematoda: Echinostomatidae) in Brazil: a morphological and molecular study

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species of trematodes belonging to the genus Drepanocephalus are intestinal parasites of piscivorous birds, primarily cormorants (Phalachrocorax spp.), and are widely reported in the Americas. During a 4-year malacological study conducted on an urban lake in Brazil, 27-collar-spined echinostome cerc...

  14. [Demography and the phenomenon of intraspecific competition of Gymnophallidae Morozov, 1955 in the second intermediate hosts (Trematoda, Digenea)].

    PubMed

    Bartoli, P

    1981-01-01

    In Gymnophallidae life-cycle, the primary host is very often a Lamelli-branch, rarely a Prosobranch; the second intermediate host is usually a Pelecypod, rarely a Prosobranch or an Annelid. The first host is generally sedentary; consequently, cercariae are not widely spread. The second intermediate host cannot explore all the environment. These characters are not in favour of the parasite. Adaptation of Gymnophallid cycle consists in a very large cercarial productivity (several millions for only one first host). The recruiting modalities of Gymnophallid larvae by the second intermediate host are not always the same. Two different mechanisms have been discovered. The first one consists of a limited recruiting. The number of metacercariae is not always growing up during the host-life. This number depends on the volume of microhabitat. The top number is reached when microhabitat is saturated. The second one consists of a illimited recruiting. The number of parasites increases during all the second host life. This peculiarity is possible because the metacercariae are permanently carried out of the microhabitat where they had settled. Parasites are indeed progressively surrounded by pallial proliferations and become enclosed inside pockets of host tissues. Thus, microhabitat is always free of parasites and infection can go on. There must be a correlation between the mechanism of the recruitment of Gymnophallid larvae by a given second host and the characteristics of the environment where the life cycle unfolds. In confined environment (almost closed lagoons, brackish ponds) the number of cercariae is very large and the recruitment observed is limited. Then, the host-life is preserved. In wide open stations, where hydrodynamism is important, cercariae are scattered. The recruitment observed is illimited. This peculiarity is probably an adaptative factor permitting concentration of larvae in the second intermediate host. PMID:7258994

  15. Experimental infection of zebra mussels Dreissena polymorpha (Mollusca:Bivalvia) by metacercariae of Echinoparyphium sp. (Platyhelminthes:Trematoda).

    PubMed

    Conn, D B; Conn, D A

    1995-04-01

    The zebra mussel Dreissena polymorpha a species recently introduced to North America from Europe, was studied to determine whether it could potentially serve as host for echinostomatid metacercariae. Forty adult mussels were collected from the St. Lawrence River and exposed to cercariae of Echinoparyphium sp. that was emerging from naturally infected Physa sp. collected from a native population in northern New York state. When necropsied 24-30 hr postexposure, 2 (5%) of the 40 harbored a metacercaria of Echinoparyphium sp. in the gonad. None of 200 control mussels collected from the same site was infected. This is the first report of any species or stage of echinostomatid from this host species worldwide. This is also the first report of an echinostomatid metacercaria from the gonad of any host species.

  16. Dissections of fresh skulls confirm low prevalence of Troglotrema acutum (Trematoda: Troglotrematidae) in German badgers (Meles meles).

    PubMed

    Heddergott, M; Frantz, A C; Jenrich, J; Müller, F

    2015-02-01

    We examined 131 European badgers Meles meles (Linnaeus, 1758) from 67 localities in central Germany for the presence of the cranial trematode Troglotrema acutum, as previous studies based on museum skulls might have underestimated the prevalence of the parasite in this host. We detected the flatworm in only three individuals that originated from the Rhoen Mountains (Thurigina and Bavaria). While the cranium of one host individual showed the lesions and the sponge-like widening of certain regions of the skullcap that are typical of a T. acutum infection, the skulls of the two remaining badgers did not show any deformations. The three badgers were infected by eight, 20, and 49 T. acutum individuals, respectively. Eggs of the trematode parasite were detected in the paranasal sinuses of two badgers. While badgers infected with T. acutum may not show any surface bone lesions, the results of the present study do not contradict the conclusion that the badger is only an accidental host of T. acutum.

  17. In vitro pairing of Echinostoma revolutum (trematoda) metacercariae and adults, and characterization of worm products involved in chemoattraction.

    PubMed

    Fried, B; Tancer, R B; Fleming, S J

    1980-12-01

    In vitro pairing studies were done on chemically excysted metacercariae and on adults of Echinostoma revolutum maintained in vitro in agar-Locke's petri dish cultures at 39 +/- 1 C for up to 24 hr. Whereas newly excysted metacercariae did not pair, both immature and mature adults showed significant pairing. Adult echinostomes confined in dialysis sacs emitted excretory-secretory (EC) products which significantly attracted single echinostome adults in vitro. Only the lipophilic fraction of ES products was found to elicit attraction. Preparative TLC analysis of adult echinostomes produced three major bands as follows: I (phospholipids); II (free sterols); and III (free fatty acids + triglycerides). When tested in vitro, only the free sterol fraction significantly attracted single adult echinostomes. TLC and GLC analyses of free sterols of E. revolutum have indicated that cholesterol is the major free sterol. PMID:7218094

  18. An integrated pipeline for next generation sequencing and annotation of the complete mitochondrial genome of the giant intestinal fluke, Fasciolopsis buski (Lankester, 1857) Looss, 1899.

    PubMed

    Biswal, Devendra Kumar; Ghatani, Sudeep; Shylla, Jollin A; Sahu, Ranjana; Mullapudi, Nandita; Bhattacharya, Alok; Tandon, Veena

    2013-01-01

    Helminths include both parasitic nematodes (roundworms) and platyhelminths (trematode and cestode flatworms) that are abundant, and are of clinical importance. The genetic characterization of parasitic flatworms using advanced molecular tools is central to the diagnosis and control of infections. Although the nuclear genome houses suitable genetic markers (e.g., in ribosomal (r) DNA) for species identification and molecular characterization, the mitochondrial (mt) genome consistently provides a rich source of novel markers for informative systematics and epidemiological studies. In the last decade, there have been some important advances in mtDNA genomics of helminths, especially lung flukes, liver flukes and intestinal flukes. Fasciolopsis buski, often called the giant intestinal fluke, is one of the largest digenean trematodes infecting humans and found primarily in Asia, in particular the Indian subcontinent. Next-generation sequencing (NGS) technologies now provide opportunities for high throughput sequencing, assembly and annotation within a short span of time. Herein, we describe a high-throughput sequencing and bioinformatics pipeline for mt genomics for F. buski that emphasizes the utility of short read NGS platforms such as Ion Torrent and Illumina in successfully sequencing and assembling the mt genome using innovative approaches for PCR primer design as well as assembly. We took advantage of our NGS whole genome sequence data (unpublished so far) for F. buski and its comparison with available data for the Fasciola hepatica mtDNA as the reference genome for design of precise and specific primers for amplification of mt genome sequences from F. buski. A long-range PCR was carried out to create an NGS library enriched in mt DNA sequences. Two different NGS platforms were employed for complete sequencing, assembly and annotation of the F. buski mt genome. The complete mt genome sequences of the intestinal fluke comprise 14,118 bp and is thus the shortest

  19. High prevalence of Schistosoma japonicum and Fasciola gigantica in bovines from Northern Samar, the Philippines.

    PubMed

    Gordon, Catherine A; Acosta, Luz P; Gobert, Geoffrey N; Jiz, Mario; Olveda, Remigio M; Ross, Allen G; Gray, Darren J; Williams, Gail M; Harn, Donald; Li, Yuesheng; McManus, Donald P

    2015-02-01

    The cause of zoonotic schistosomiasis in the Philippines is Schistosoma japonicum, which infects up to 46 mammalian hosts, including humans and bovines. In China, water buffaloes have been identified as major reservoir hosts for schistosomiasis japonica, contributing up to 75% of human transmission. In the Philippines, water buffaloes (carabao; Bubalus bubalis carabanesis) have, historically, been considered unimportant reservoirs. We therefore revisited the possible role of bovines in schistosome transmission in the Philippines, using the recently described formalin-ethyl acetate sedimentation (FEA-SD) technique and a qPCR assay to examine fecal samples from 153 bovines (both carabao and cattle) from six barangays in Northern Samar. A high prevalence of S. japonicum was found using qPCR and FEA-SD in both cattle (87.50% and 77.08%, respectively) and carabao (80.00% and 55.24%, respectively). The average daily egg output for each bovine was calculated at 195,000. High prevalence and infection intensity of F. gigantica was also found in the bovines by qPCR and FEA-SD (95.33% and 96.00%, respectively). The identification of bovines as major reservoir hosts for S. japonicum transmission suggests that bovine treatment and/or vaccination, as one becomes available, should be included in any future control program that aims to reduce the disease burden due to schistosomiasis in the Philippines.

  20. Evaluation of cases with Fasciola hepatica infection: experience in 6 children

    PubMed Central

    Tezer, Hasan; Yuksek, Saliha Kanik; Parlakay, Aslinur Özkaya; Gülhan, Belgin; Tavil, Betül; Tunç, Bahattin

    2013-01-01

    Objective To evaluate 6 pediatric cases diagnosed as fascioliasis. Methods Patients diagnosed as fascioliasis are retrospectively interpreted. Results Herein 6 pediatric cases having a mean age of (12.7±3.1) years with different complaints including fever, weakness, paleness, sweating, cough, headache, rectal bleeding, abdominal pain, and abdominal distension admitting to our hospital are evaluated. Conclusions Diagnosis of fascioliasis needs a high index of suspicion supported by stool microscopy, serology, imaging procedures, and in difficult cases even liver biopsy. Triclabendazole resistance should be kept in mind in unresponsive cases.

  1. The effect of potassium permanganate and sodium dichloroisocyanurate on metacercariae of Fasciola gigantica.

    PubMed

    el-Zawawy, Lobna A; Ali, Safeya M; Allam, Sonia R

    2003-04-01

    Batches of encysted metacercariae of F. gigantica, adhered to transparent polyethylene sheets, were treated with KMnO4, while others were treated with NaDCC at specific concentrations and exposure times. Assessment of the effects was carried out by the detached percentage and viable metacercariae and by scanning electron microscope (SEM) ultrastructure changes. In addition, their effects on leaves of green vegetables were reported. The results showed that all metacercariae were detached and were dead by exposure to KMnO4 (96%) and NaDCC (100%) were detached from the polyethylene sheets. SEM showed that the deformities in the metacercariae soaked in NaDCC were more severe than those dipped in KMnO4. However, neither KMnO4 nor NaDCC affected the consistency, color, taste or flavor of the vegetables' leaves. The two disinfectants particularly NaDCC, proved to be safe, effective against the encysted metacercariae with no side-effects. PMID:12739819

  2. Crowding effect on adult growth, pre-patent period and egg shedding of Fasciola hepatica.

    PubMed

    Valero, M A; De Renzi, M; Panova, M; Garcia-Bodelon, M A; Periago, M V; Ordoñez, D; Mas-Coma, S

    2006-10-01

    Fascioliasis pathogenesis depends on fluke burden. In human hyperendemic zones, individual infection intensities reach very high levels and the majority of infected subjects should be in the advanced chronic phase. The rat model offers a useful approach for pathological research in the advanced chronic period. The influence of infection intensity per rat on fluke development, pre-patent period and egg shedding (eggs/g faeces/worm) was analysed in 3 groups (I: 1-3 worms/rat; II: 4-6; III: 7-9). Ontogenetic trajectories of fluke body measures followed a logistic model. Results showed that when the burden increases, the maximum values of fluke measures decrease. The crowding effect is manifest when fluke measures approximate their maximums in the advanced chronic stage. The pre-patent period and egg production decrease when the burden increases. This means that measurements of eggs per gramme of faeces tend to underestimate the fluke burden. The present study demonstrates how to quantify the fascioliasis experimental rat model crowding effect on adult growth, pre-patent period and egg production. This quantification may be of great interest in epidemiological studies and in experimental research on the in vivo actions of different anthelminthic drugs and vaccines, pathology, immunology and resistance studies.

  3. Comparative characterization of microRNAs from the liver flukes Fasciola gigantica and F. hepatica.

    PubMed

    Xu, Min-Jun; Ai, Lin; Fu, Jing-Hua; Nisbet, Alasdair J; Liu, Qing-You; Chen, Mu-Xin; Zhou, Dong-Hui; Zhu, Xing-Quan

    2012-01-01

    MicroRNAs (miRNAs) are key regulators of gene expression at the post-transcription level. The present study specifically explored and compared the miRNA expression profiles of F. gigantica and F. hepatica using an integrated sequencing and bioinformatics platform and quantitative real-time PCR. Nineteen and 16 miRNA candidates were identified from F. gigantica and F. hepatica, respectively. The two parasites shared 11 miRNAs, with 8 also showing similarity to miRNAs of Schistosoma japonicum. Another 8 miRNAs were identified as F. gigantica-specific and 5 as F. hepatica-specific, most of which were novel. Predicted target analysis with 11465 mRNA and EST sequences of F. hepatica and F. gigantica revealed that all of the miRNAs had more than one target, ranging from 2 to 398 with an average of 51 targets. Some functions of the predicted targets were only found in F. gigantica, such as "transcription regulator", while some others were only found in F. hepatica, such as "reproduction" and "response to stimulus", indicating the different metabolism and gene regulation patterns of the two parasites. The present study represents the first global comparative characterization of miRNA expression profiles of F. gigantica and F. hepatica, which has provided novel valuable resources for a better understanding of the two zoonotic trematodes.

  4. Association of Fasciola hepatica Infection with Liver Fibrosis, Cirrhosis, and Cancer: A Systematic Review

    PubMed Central

    Machicado, Claudia; Machicado, Jorge D.; Maco, Vicente; Terashima, Angelica; Marcos, Luis A.

    2016-01-01

    Background Fascioliasis has been sporadically associated with chronic liver disease on previous studies. In order to describe the current evidence, we carried out a systematic review to assess the association between fascioliasis with liver fibrosis, cirrhosis and cancer. Methodology and Principal Findings A systematic search of electronic databases (PubMed, LILACS, Scopus, Embase, Cochrane, and Scielo) was conducted from June to July 2015 and yielded 1,557 published studies. Among 21 studies that met inclusion and exclusion criteria, 12 studies explored the association of F. hepatica with liver fibrosis, 4 with liver cirrhosis, and 5 with cancer. Globally these studies suggested the ability of F. hepatica to promote liver fibrosis and cirrhosis. The role of F. hepatica in cancer is unknown. Given the heterogeneity of the studies, a meta-analysis could not be performed. Conclusions Future high-quality studies are needed to determine the role of F. hepatica on the development of liver fibrosis, liver cirrhosis, and cancer in humans. PMID:27681524

  5. The mitochondrial genome of Paragonimus westermani (Kerbert, 1878), the Indian isolate of the lung fluke representative of the family Paragonimidae (Trematoda)

    PubMed Central

    Biswal, Devendra K.; Chatterjee, Anupam

    2014-01-01

    Among helminth parasites, Paragonimus (zoonotic lung fluke) gains considerable importance from veterinary and medical points of view because of its diversified effect on its host. Nearly fifty species of Paragonimus have been described across the globe. It is estimated that more than 20 million people are infected worldwide and the best known species is Paragonimus westermani, whose type locality is probably India and which infects millions of people in Asia causing disease symptoms that mimic tuberculosis. Human infections occur through eating raw crustaceans containing metacercarie or ingestion of uncooked meat of paratenic hosts such as pigs. Though the fluke is known to parasitize a wide range of mammalian hosts representing as many as eleven families, the status of its prevalence, host range, pathogenic manifestations and its possible survivors in nature from where the human beings contract the infection is not well documented in India. We took advantage of the whole genome sequence data for P. westermani, generated by Next Generation Sequencing, and its comparison with the existing data for the P. westermani for comparative mt DNA phylogenomic analyses. Specific primers were designed for the 12 protein coding genes with the aid of existing P. westermani mtDNA as the reference. The Ion torrent next generation sequencing platform was harnessed to completely sequence the mitochondrial genome, and applied innovative approaches to bioinformatically assemble and annotate it. A strategic PCR primer design utilizing the whole genome sequence data from P. westermani enabled us to design specific primers capable of amplifying all regions of the mitochondrial genome from P. westermani. Assembly of NGS data from libraries enriched in mtDNA sequence by PCR gave rise to a total of 11 contigs spanning the entire 14.7 kb mt DNA sequence of P. westermani available at NCBI. We conducted gap-filling by traditional Sanger sequencing to fill in the gaps. Annotation of non-protein coding genes successfully identified tRNA regions for the 24 tRNAs coded in mtDNA and 12 protein coding genes. Bayesian phylogenetic analyses of the concatenated protein coding genes placed P. westermani within the family Opisthorchida. The complete mtDNA sequence of P. westermani is 15,004 base pairs long; the lung fluke is the major etiological agent of paragonimiasis and the first Indian representative for the family Paragonimidae to be fully sequenced that provides important genetic markers for ecological, population and biogeographical studies and molecular diagnostic of digeneans that cause trematodiases. PMID:25165620

  6. [Infestation of littoral molluscs of Dal'nie Zelentsy Bay by parthenitae of the eider parasite Microphallus pygmaeus (Levinsen, 1881) (Trematoda: Microphallidae)].

    PubMed

    Podlipaev, S A

    1975-01-01

    The mollusks Littorina saxatilis, L. obtusata, Solariella obscura, Cingula aculeus, Hydrobia ulvae, Margarites helicina and M. groenlandica serve as hosts of the parthenites of Microphallus pygmaeus in eastern Murman. Over a period of 20 years the infection rate of the mollusks has increased greatly that is associated with the increasing number of definitive hosts. Maximum infection rate was recorded in the end of July--August. The infection rate of littorins within this period was about 90 per cent. PMID:130602

  7. Where are the South American freshwater turtle blood flukes (Trematoda: Spirorchiidae)? The first morphological and molecular analysis of spirorchiid cercariae from freshwater snails in Brazil.

    PubMed

    Pinto, Hudson Alves; de Melo, Alan Lane; Brant, Sara Vanessa

    2015-12-01

    Trematodes belonging to the family Spirorchiidae are blood parasites mainly of turtles with a worldwide distribution. These flukes were recently reported in some marine turtles from South America, where the occurrence of spirorchiids in freshwater definitive and intermediate hosts is so far unknown. In the present study, three morphotypes of brevifurcate apharyngeate distome cercariae found in freshwater molluscs from an urban reservoir in Brazil were used for morphological and molecular (nuclear 28S rDNA) evaluation. Two morphotypes of cercariae, probably congeneric species, were found in 12/17,465 specimens of Biomphalaria spp. and differ from each other by body size and sequences (0.1%). They present morphology similar to North American freshwater spirorchiids (Spirorchis spp.), however surprisingly molecular data reveals that these lineages are more closely related to marine spirorchiids. A third species found in 2/777 Pomacea sp. differs morphologically from all previously described spirorchiid cercariae and genetically from spirorchiids with available sequences (16-19%), grouping in the phylogenetic tree with freshwater North American species. This is the first report of freshwater spirorchiids in South America and the first molecular confirmation of the involvement of a caenogastropod in the life cycle of spirorchiids.

  8. A species pair of Bivesicula Yamaguti, 1934 (Trematoda: Bivesiculidae) in unrelated Great Barrier Reef fishes: implications for the basis of speciation in coral reef fish trematodes.

    PubMed

    Trieu, Nancy; Cutmore, Scott C; Miller, Terrence L; Cribb, Thomas H

    2015-07-01

    Combined morphological and molecular analysis shows that a species of Bivesicula Yamaguti, 1934 from four species of Apogonidae Günther [Nectamia fusca (Quoy & Gaimard), Ostorhinchus angustatus (Smith & Radcliffe), O. cookii (Macleay) and Taeniamia fucata (Cantor)] on the Great Barrier Reef is morphologically similar to, but clearly distinct from B. unexpecta Cribb, Bray & Barker, 1994 which infects a sympatric pomacentrid, Acanthochromis polyacanthus (Bleeker). Bivesicula neglecta n. sp. is proposed for the form from apogonids. Novel ITS2 rDNA sequences generated for the two species differ at just one consistent base position, implying that the two species are closely related. The combination of their close relationship, high but distinct specificity and co-occurrence suggests that speciation was driven by a recent host switching event enabled by similar dietary ecomorphology.

  9. [Microphallidae Travassos, 1920 (Trematoda). XXXIII. Apropos of 11 species described or collected by P. Oschmarin in the Far East (author's transl)].

    PubMed

    Deblock, S

    1975-01-01

    The collection of the Microphallidae collected by P. Oschmarin includes the following species: 1) Maritrema magnicirrus, M. gratiosum, Maritrema sp.; 2) Basantisia halcyonae, B. longa, B. ramai -- Belopolskiella prolecithum; 3) Microphallus oviformis; 4) Levinseniella acanthophalla, L. bucephalae tringae; 5) Spiculotrema littoralis. The writer describes and illustrates the anatomy of several of those species; consequently he proposes the following changes of systematic: 1) Maritrema gratiosum sensu Oschmarin is a new species: Maritrema (Maritrema) chiriacae; the Maritrema sp. could be M. sachalinicum Schumakowitsch; 2) The genus Belopolskiella and Basantisia are synonymous; the species prolecithum and longa are synonymous. Besides B. prolecithum and B. tamsuiensis could be synonymous but it is not proved; 3) L. acanthophalla and L. bucephalae tringae are both synonimisided with L. bucephalae Yamaguti, 1939.

  10. [Microphallus kurilensis sp. nov., a new species of microphallids from the pygmaeus species group (Trematoda, Microphallidae) from the coastal areas of Okhotsk and Bering Seas].

    PubMed

    Galaktionov, K V; Regel', K V; Atrashkevich, G I

    2010-01-01

    The pygmaeus-species group is composed of close related species from the genus Microphallus in which metacercariae develop inside daughter sporocysts without encystment. Infection of periwinkles Littorina (Neritremna) spp. with intramolluscan stages of a new species of this group (Microphallus kurilensis sp. nov.) was recorded on the coasts of Sakhalin and Kuril islands, north of the Sea of Okhotsk and Chukchi Peninsula (the Bering Sea). Application of molecular methods allowed us to establish that M. kurilensis metacercariae are conspecific with one of the morphotypes of microphallid adults obtained from the intestine of the Pacific common eider (Somateria mollissima v-nigrum), which was shot in the north of the Sea of Okhotsk (Galaktionov, Olson, and Blasco-Costa, in press). The adults of the same morphotype were recorded in the Pacific common eider from the northwestern part of the Bering Sea (Chukchi Peninsula). In the course of experimental infection of the slaty-backed gull Larus schistisagus chicks with metacercariae of M. kurilensis, few microphallid adults were obtained. These adults were identical in their morphology with specimens of the microphallid morphotype from the Pacific common eider, which had been identified as M. kurilensis based on molecular data. Morphological description of metacercaria and adult of M. kurilensis and list of their differences from the same developmental stages of other species from pygmaeus-group are provided. It is concluded that M. kurilensis is transmitted in the host system including periwinkle Littorina (Neritrema) and seaducks (predominately, Pacific common eider). Most probably, distribution of M. kurilensis is not limited by the north Asiatic coast but expanded to the North American coast of the Pacific Ocean. PMID:21427958

  11. Evaluation of the mitochondrial system in the gonad-digestive gland complex of Biomphalaria glabrata (Mollusca, Gastropoda) after infection by Echinostoma paraensei (Trematoda, Echinostomatidae).

    PubMed

    Tunholi, Victor Menezes; Tunholi-Alves, Vinícius Menezes; Santos, Anderson Teixeira; Garcia, Juberlan da Silva; Maldonado, Arnaldo; da-Silva, Wagner Seixas; Rodrigues, Maria de Lurdes de Azevedo; Pinheiro, Jairo

    2016-05-01

    The effect of infection by Echinostoma paraensei on the mitochondrial physiology of Biomphalaria glabrata was investigated after exposure to 50 miracidia. The snails were dissected one, two, three and four weeks after infection for collection and mechanical permeabilization of the gonad-digestive gland (DGG) complex. The results obtained indicate that prepatent infection by this echinostomatid fluke significantly suppresses the phosphorylation state (respiratory state 3) and basal oxygen consumption of B. glabrata, demonstrating that the infection reduces the ability of the intermediate host to carry out aerobic oxidative reactions. Additionally, relevant variations related to the uncoupled mitochondrial (state 3u) of B. glabrata infected by E. paraensei were observed. Four weeks after exposure, a significant reduction in mitochondrial oxygen consumption after addition of ADP (3.68±0.26pmol O2/mg proteins) was observed in the infected snails in comparison with the respective control group (5.14±0.25). In the uncoupled state, the infected snails consumed about 62% less oxygen than the infected snails (7.87±0.84pmol O2/mg proteins) in the same period. These results demonstrate a reduction in oxidative decarboxylation rate of the tricarboxylic acid cycle and faster anaerobic degradation of carbohydrates in the infected snails. The possible mechanisms that explain this new metabolic condition in the infected organisms are discussed. PMID:27079167

  12. Influence of Ribeiroia ondatrae (Trematoda: Digenea) infection on limb development and survival of northern leopard frogs (Rana pipiens): effects of host stage and parasite-exposure level

    USGS Publications Warehouse

    Schotthoefer, Anna M.; Koehler, Anson V.; Meteyer, Carol U.; Cole, Rebecca A.

    2003-01-01

    Recent evidence suggests that infection by larvae of the trematode Ribeiroia ondatrae accounts for a significant proportion of limb malformations currently observed in amphibian populations of North America. However, the effects of R. ondatrae infection on northern leopard frogs (Rana pipiens), one of the species most frequently reported with malformations, have not been adequately explored. Moreover, the risk factors associated with R. ondatrae-induced malformations have not been clearly identified. We examined the effects of timing of infection on tadpole survival and limb development. Rana pipiens tadpoles were individually exposed to R. ondatrae cercariae at the pre-limb-bud (Gosner stages 24 and 25), limb-bud (Gosner stages 27 and 28), or paddle (Gosner stages 31–33) stages of development and monitored through metamorphosis. The effects of infection were stage-specific. Infections acquired at the pre-limb-bud stage resulted in a high mortality rate (47.5–97.5%), whereas tadpoles infected at the limb-bud stage displayed a high malformation rate (16% overall), and the magnitude of effects increased with the level of exposure to cercariae. In contrast, infections acquired at the paddle stage had no effect on limb development or tadpole survival, which suggests that the timing of R. ondatrae infection in relation to the stage structure of tadpole populations in the wild is an important determinant of the degree to which populations are affected by R. ondatrae.

  13. Distribution and seasonal abundance of trematode parasites (Trematoda: Allocreadiidae: Crepidostomum spp.) in burrowing mayfly nymphs (Ephemeroptera: Ephemeridae: Hexagenia spp.) from connecting rivers of the Laurentian Great Lakes

    USGS Publications Warehouse

    Schloesser, Don W.

    2005-01-01

    Burrowing-mayfly nymphs such as Hexagenia spp. have been used extensively in North America and Europe as a biomonitoring tool to indicate mesotrophic water quality, yet infestation by associated parasites has not been well documented. We performed laboratory analysis of archived samples of Hexagenia spp. nymphs collected in 1985 and 1986 to provide base-line data on the distribution (1985) and seasonal infestation (1986) of the trematode parasite Crepidostomum spp. in Hexagenia spp. nymphs in connecting rivers between Lakes Superior and Erie of the Laurentian Great Lakes. In May and June 1985, frequency of occurrence of metacercariae was widely distributed throughout the connecting rivers (63% of 203 stations with nymphs), except in areas where nymph densities were relatively low (i.e.,a?Y69 nymphs/mA?). Distribution was probably underestimated in the present study because of low probability (mean = 31%, range = 0-57%) of detecting infestation in a small number of collected nymphs (a??10) at nymph densities a??69/mA?. In 1986, seasonal infestation between April and October occurred in 3.3% (627) of 18696 nymphs. Overall prevalence, mean intensity, and mean abundance of parasites at one station in the St. Marys River indicate parasite transmission occurred between June and September. This period of transmission is dependent on the life-cycle of the parasite. In addition, the life-cycle of Hexagenia spp. determines which annual cohort of nymphs is infested and therefore, the duration of infestation. Although, no impacts of infestation on Hexagenia spp. nymphs were observed in the present study, infestation intensities were high enough (a?Y25 metacercariae per nymph) at one station in the St. Marys River to potentially cause tissue damage in a high proportion (53%) of infested nymphs.

  14. Urotrema shirleyae n. sp. (Trematoda: Digenea: Urotrematidae) in Norops oxylophus and N. cupreus (Squamata: Iguania: Polychrotidae) from the Area de Conservación Guanacaste, Costa Rica.

    PubMed

    Zamparo, David; Brooks, Daniel R; Tkach, Vasyl

    2005-06-01

    A new species of Urotrema inhabits Norops oxylophus and Norops cupreus from the Area de Conservación Guanacaste, Costa Rica. The new species is most similar to Urotrema scabridum but differs by having a relatively longer esophagus, at least 10% of the total body length versus 4-8% in U. scabridum; an oral sucker width: ventral sucker width ratio less than 1 : 1 (averaging 1 : 0.76) versus 1 : 1 or greater; an oral sucker width : pharyngeal width ratio less than 1 : 0.5 (averaging 1 : 0.42) versus greater than 1 :0.5; and by having many transversely oriented posttesticular uterine loops versus a few vertically oriented posttesticular uterine loops.

  15. Tinamutrema canoae n. gen. et n. sp. (Trematoda: Digenea: Strigeiformes: Brachylaimidae) in Crypturellus cinnamomeus (Aves, Passeriformes, Tinamidae) from the Area de Conservación Guanacaste, Costa Rica.

    PubMed

    Zamparo, David; Brooks, Daniel R; Causey, Douglas

    2003-08-01

    We propose Tinamutrema as a new genus for Brachylaima centrodes (Braun, 1901) Dollfus, 1935 and for T. canoae, as a new species inhabiting tinamus in the Area de Conservación Guanacaste, Costa Rica. Specimens from Costa Rica resemble B. centrodes in having an elongate body, pretesticular genital pore and terminal genitalia, intercecal uterine loops occupying all available space between the anterior testis and the intestinal bifurcation, an oral sucker width:pharynx width ratio of approximately 1:0.55, an oral sucker:ventral sucker width ratio of approximately 1:1, and vitelline follicles extending into the forebody closer to the pharynx than to the anterior margin of the ventral sucker and by living in the cloaca. They differ from B. centrodes in having vitelline follicles that do not extend as far anteriorly as those in B. centrodes, which extend anteriorly to the level of the anteriormost extent of the cecal "shoulders," dense tegumental spination as opposed to sparse or no spination, relatively smaller cirrus with fewer spines, longer and more sinous pars prostatica, and forebody averaging 36% of total body length (TBL) as opposed to 42% TBL. Both species differ from other members of the Brachylaimidae in possessing a spinose cirrus and a cirrus sac containing both the cirrus and the pars prostatica. Preliminary phylogenetic assessment suggests that these traits are plesiomorphic, and thus the species are basal to the rest of the Brachylaimidae, whose diagnosis we emend accordingly.

  16. The identification and characteristics of Echinoparyphium rubrum (Cort. 1914) new comb. (Trematoda, Echinostomatidae) based on experimental evidence of the life cycle

    USGS Publications Warehouse

    Kanev, I.; Sorensen, R.; Sterner, M.; Cole, R.; Fried, B.

    1998-01-01

    The life cycle of Echinoparyphium rubrum (Cort, 1914) comb. n. has been completed experimentally. All of the developmental stages egg, miracidium, sporocyst, mother and daughter rediae, cercaria, metacercaria, and adult were examined and described. The miracidia infected freshwater snails of the genus Physa , P. gyrina and P. occidentalis. Attempts to infect snails of the genera Lymnaea, L. auricularis, L. peregra, L. truncatula and Bulinus, B. truncatus failed. Cercariae infected various pulmonate and prosobranch freshwater snails, mussels, frogs, water turtles and planarians. The adults developed in the small intestine of birds and mammals. The identity and major characteristics of Echinoparyphium rubrum are discussed. Synonyms of E. rubrum are Cercaria rubra Cort, 1914; Cercaria biflexa Faust, 1917; Cercaria chisolenata Faust, 1918; Echinostoma callawayensis Barker et Noll, 1915; Echinostoma revolutum of Johnson (1920); Echinoparyphium elegans of Cannon (1938), of Bain and Trelfall (1977), of Mahoney and Trelfall (1977); and Echinoparyphium recurvatum of Jilek (1977), Harley (1972), Sankurathri and Holmes (1976). Comparisons are made between E. rubrum and its 43-collar-spined allies: E. flexum from North America, E. cinctum from Europe, E. dunni from Asia and E. elegans from Africa.

  17. Central European parasitic flatworms of the family Renicolidae Dollfus, 1939 (Trematoda: Plagiorchiida): molecular and comparative morphological analysis rejects the synonymization of Renicola pinguis complex suggested by Odening.

    PubMed

    Heneberg, Petr; Sitko, Jiljí; Bizos, Jiří; Horne, Elizabeth C

    2016-10-01

    The Renicolidae are digenean parasites of piscivorous and molluscivorous birds. Although they exhibit few morphological autapomorphies and are highly variable, the numerous suggested re-classifications within the family have never been supported by any molecular analyses. We address the possible synonymization of species within the Renicola pinguis complex suggested previously by Odening. We provide and analyse sequences of two nuclear (ITS2, 28S rDNA) and two mitochondrial (CO1, ND1) DNA loci of central European species of the Renicolidae, namely Renicola lari, Renicola pinguis and Renicola sternae sp. n., and we also provide first sequences of Renicola sloanei. The combined molecular and comparative morphological analysis confirms the previously questioned validity of the three Renicola spp. of highly similar morphology, which display strict niche separation in terms of host specificity and selectivity. We identify two previously unreported clades within the genus Renicola; however, only one of them is supported by the analysis of adult worms. We also provide comparative measurements of the three examined closely related central European renicolids, and describe the newly proposed tern-specialized species Renicola sternae sp. n., which was previously repeatedly misidentified as Renicola paraquinta. Based on the extensive dataset collected in 1962-2015, we update the host spectrum of Renicolidae parasitizing central European birds (Renicola bretensis, R. lari, Renicola mediovitellata, R. pinguis, Renicola secunda and R. sternae sp. n.) and discuss their host-specific prevalence and intensity of infections. PMID:27356772

  18. [A comparative study of excretory and sensory apparatus of two cercariae of Diplodiscus parasites of European and African Amphibians (Trematoda, Diploidiscidae) (author's transl)].

    PubMed

    Bayssade-Dufour, C; Albaret, J L; Grabda-Kazubska, B; Kulo, S D

    1978-01-01

    The comparative study of the excretory system provides new data on the number of flame cells of two cercariae of Diplodiscus from Poland and Togo but does not allow to distinguish them. On the other hand, the comparison of the superficial sensory apparatus shows significant differences. The cercariae from Togo differ from these of Poland by a much lower number of all the cephalic papillae and papillae of cycle Cv, a higher number of acetabular papillae, and the presence of 9 to 12 pairs of dorsal papillae in the median part of the tail. Therefore the African Diplodicus is not D. subclavatus. We identify it as D. fischthalicus. PMID:754617

  19. A redescription of Lobatostoma kemostoma (MacCallum & MacCallum, 1913) (Trematoda: Aspidogastrea) from the florida pompano fish Trachinotus carolinus (Linnaeus, 1766) off the Brazilian coast.

    PubMed

    Alves, P V; Borges, J N; Santos, C P; Luque, J L

    2015-05-01

    A redescription of the aspidogastrean trematode Lobatostoma kemostoma (MacCallum & MacCallum, 1913) is provided based on examination of type material, voucher specimens available at institutional collections and newly collected specimens from the type host, Trachinotus carolinus (Linnaeus, 1766), off the Angra dos Reis, Brazil. Partial sequence of its 28S rDNA is given. Lobatostoma kemostoma can be distinguished from the other eight species of the genus by: (1) the hindbody being longer than the ventral disc; and (2) the cephalic lobes having the same shape and size. Features observed for the first time include the number and detailed arrangement of alveoli on the ventral disc, the terminal genitalia and the ovarian complex. New illustrations and morphometric data are given. In the present survey the host specificity in the genus, the validity of some morphological features and the occurrence of an Indian species of Lobatostoma from off the Brazilian coast are discussed. A lectotype of L. kemostoma from the type series deposited at United States National Parasite Collection is also designated.

  20. Experimental verification of the metacercarial stage of Rhipidocotyle lepisostei (Trematoda: Bucephalidae) with notes on the natural occurrence of its adult stage in gars in Texas and Virginia.

    PubMed

    Wardle, W J

    1990-04-01

    Bucephalid metacercariae found encysted in the caudal fin rays of striped mullet (Mugil cephalus) were fed to bucephalid-free spotted gars (Lepisosteus oculatus) in the laboratory. Gravid specimens of Rhipidocotyle lepisostei were recovered from the intestinal tracts of 2 of 3 gars from 7 to 28 days later. Further attempts to complete the entire life cycle were unsuccessful. Natural infections of adult R. lepisostei were found in the type host, the alligator gar Lepisosteus spatula, from Galveston Bay, Texas, and in the long-nose gar, Lepisosteus osseus, from the York River, Virginia. This report confirms the identity of the metacercarial stage in the mullet as R. lepisostei and extends the known range of this parasite from the Louisiana and Texas area to the U.S. Atlantic coast. PMID:2319436

  1. Experimental verification of the metacercarial stage of Rhipidocotyle lepisostei (Trematoda: Bucephalidae) with notes on the natural occurrence of its adult stage in gars in Texas and Virginia.

    PubMed

    Wardle, W J

    1990-04-01

    Bucephalid metacercariae found encysted in the caudal fin rays of striped mullet (Mugil cephalus) were fed to bucephalid-free spotted gars (Lepisosteus oculatus) in the laboratory. Gravid specimens of Rhipidocotyle lepisostei were recovered from the intestinal tracts of 2 of 3 gars from 7 to 28 days later. Further attempts to complete the entire life cycle were unsuccessful. Natural infections of adult R. lepisostei were found in the type host, the alligator gar Lepisosteus spatula, from Galveston Bay, Texas, and in the long-nose gar, Lepisosteus osseus, from the York River, Virginia. This report confirms the identity of the metacercarial stage in the mullet as R. lepisostei and extends the known range of this parasite from the Louisiana and Texas area to the U.S. Atlantic coast.

  2. Species of Apatemon Szidat, 1928 and Australapatemon Sudarikov, 1959 (Trematoda: Strigeidae) from New Zealand: linking and characterising life cycle stages with morphology and molecules.

    PubMed

    Blasco-Costa, Isabel; Poulin, Robert; Presswell, Bronwen

    2016-01-01

    Species of Apatemon Szidat, 1928 and Australapatemon Sudarikov, 1959 are reported from New Zealand for the first time, and their life cycles are resolved using molecular sequence data (28S and ITS rDNA regions and mitochondrial COI). The metacercaria of Apatemon sp. 'jamiesoni' ex Gobiomorphus cotidianus and its cercaria ex Potamopyrgus antipodarum are described in detail. Its adult, found in Anas platyrhynchos and Phalacrocorax punctatus, is identified by molecular sequence data. Apatemon sp. 'jamiesoni' uses a different species of snail host, exhibits consistent differences in the genetic markers examined and its single described adult differs from known species so as to be considered distinct, but its formal description awaits additional adult specimens. Australapatemon niewiadomski n. sp. is described from Anas platyrhynchos. It is distinguished morphologically by the absence of a ringnapf and its overall smaller size compared to most other Australapatemon spp. except Au. magnacetabulum and Au. minor, which are smaller in nearly all features than the new species. Au. niewiadomski n. sp. metacercaria and its intermediate host (Barbronia weberi) are identified via matching of molecular sequence data. The status of Apatemon and Australapatemon as distinct genera is confirmed based on their respective monophyly, and genetic divergence between them is comparable to other well-established genera in the Strigeidae. The diagnosis of Australapatemon is emended. Life history data, accurate patterns of host specialisation and distribution, alongside concurrent molecular and morphological evidence would be useful for an integrative taxonomical approach towards the elucidation of species diversity in this group.

  3. Discovery of intermediate hosts for two species of blood flukes Cardicola orientalis and Cardicola forsteri (Trematoda: Aporocotylidae) infecting Pacific bluefin tuna in Japan.

    PubMed

    Shirakashi, Sho; Tani, Kazuki; Ishimaru, Katsuya; Shin, Sang Phil; Honryo, Tomoki; Uchida, Hiro'omi; Ogawa, Kazuo

    2016-04-01

    Fish blood flukes (Aporocotylidae) are important pathogens of farmed finfish around the world. Among them, Cardicola spp. infecting farmed tuna are considered to be serious threats to tuna farming and have received tremendous attention. We conducted periodical samplings at a tuna farming site in Japan between January and May, 2015 to determine the life cycle of Cardicola spp. We collected over 4700 terebellid polychaetes from ropes, floats and frames of tuna culture cages and found nearly 400 infected worms. Sporocysts and cercariae found in Nicolea gracilibranchis were genetically identified as Cardicola orientalis by 28S and ITS2 ribosomal DNA sequences. This was the first discovery of the intermediate host for this parasite species. Infection prevalence and the abundance of N. gracilibranchis significantly varied between sampling points and the highest number of infected terebellids were collected from ropes. We also demonstrated morphologically and molecularly that asexual stages found in a single Amphitrite sp. (Terebellidae) and adult worms isolated from farmed juvenile tuna were Cardicola forsteri. This is the first report of C. forsteri in Pacific bluefin tuna (PBT) Thunnus orientalis in Japan. Our results demonstrated that all three species of Cardicola orientalis, C. forsteri and Cardicola opisthorchis exist in Japanese farmed PBTs and that they all use terebellid polychaetes as the intermediate hosts.

  4. Transversotrema Witenberg, 1944 (Trematoda: Transversotrematidae) from inshore fishes of Australia: description of a new species and significant range extensions for three congeners.

    PubMed

    Cutmore, Scott C; Diggles, Ben K; Cribb, Thomas H

    2016-09-01

    Four transversotrematid trematodes are reported from commercial teleost species in Australian waters. Transversotrema hunterae n. sp. is described from three species of Sillago Cuvier (Sillaginidae) from Moreton Bay, south-east Queensland. Molecular characterisation using ITS2 rDNA confirmed this stenoxenic specificity of Transversotrema hunterae n. sp., with identical sequence data from Sillago maculata Quoy & Gaimard, S. analis Whitley and S. ciliata Cuvier. Phylogenetic analysis, based on 28S rDNA data, demonstrates that T. hunterae n. sp. belongs to the 'Transversotrema licinum clade' and is most closely related to Transversotrema licinum Manter, 1970 and T. polynesiae Cribb, Adlard, Bray, Sasal & Cutmore, 2014, with the three species forming a well-supported clade in all analyses. We extend the known host and geographical ranges of three previously described Transversotrema species, T. licinum, T. elegans Hunter, Ingram, Adlard, Bray & Cribb, 2010 and T. espanola Hunter & Cribb, 2012. The new records represent significant range extensions for the three species and permit further examination of the patterns of biogeographical distribution in Australian waters. Host-specificity of Transversotrema species is examined, and the degree to which morphological analysis can inform taxonomic studies of this group is discussed. PMID:27522364

  5. Infrapopulations of Gyliauchen volubilis Nagaty, 1956 (Trematoda: Gyliauchenidae) in the rabbitfish Siganus rivulatus (Teleostei: Siganidae) from the Saudi coast of the Red Sea

    PubMed Central

    Al-Jahdali, M.O.

    2012-01-01

    In hermaphroditic helminth parasites, infrapopulation size or mating group size mostly affects some processes acting within the infrapopulation. Here, 30 natural infrapopulations (12-154 individuals) of the intestinal trematode Gyliauchen volubilis Nagaty, 1956 from the fish Siganus rivulatus consisting of newly excysted juveniles, immature and mature worms were found distributed in a well-defined fundamental niche (anterior 40 % of the intestine). In small infrapopulations, all stages of the parasite were alive. In larger infrapopulations, differential mortality was only and consistently observed among newly excysted juveniles, and gradually increased to include most or all juveniles in the largest infrapopulations. Among mature worms, the mean worm length seemed unaffected by the infrapopulation size. However, the ratio mean testis size-mean ovary size, a reliable indicator of resource allocation to the male function and of opportunities for crossfertilization, significantly increased with mating group size. In small infrapopulations, all stages of the parasite were scattered along the niche, and never seen in mating pairs (possibly reproduced by selffertilization). In larger infrapopulations, newly excysted juveniles and immature worms were scattered along the anterior two thirds of the niche, while mature worms were constantly found aggregated in its posterior third (narrow microhabitat), where some were arranged in mating pairs. The probability of mating reciprocally or unilaterally was dependent on body size. The mean number of uterine eggs per worm significantly decreased and their mean sizes significantly increased with mating group size. The results are statistically significant and suggest that infrapopulation self-regulation is greatly associated with its size. PMID:22910665

  6. Biogeography of tropical Indo-West Pacific parasites: a cryptic species of Transversotrema and evidence for rarity of Transversotrematidae (Trematoda) in French Polynesia.

    PubMed

    Cribb, Thomas H; Adlard, Robert D; Bray, Rodney A; Sasal, Pierre; Cutmore, Scott C

    2014-04-01

    We sought transversotrematid trematodes from French Polynesian fishes by examining 304 individual scaled fishes of 53 species from seven families known to harbour the family elsewhere. A single species was found at two locations in the Tuamotus Archipelago on two species of Chaetodontidae (Chaetodon auriga and Chaetodon ephippium) and one species of Lutjanidae (Lutjanus gibbus). The species closely resembles Transversotrema borboleta Hunter & Cribb, 2012 from chaetodontids and lutjanids of the northern Great Barrier Reef (GBR) but differs from it consistently in 8 base positions of ITS2 rDNA. This level of variation exceeds that between some clearly morphologically distinct pairs of species of Transversotrema and the form from French Polynesia is thus interpreted as a distinct, though cryptic, species and named Transversotrema polynesiae n. sp. The new species forms part of a complex of species, here characterised as the T. borboleta complex, associated with chaetodontids and lutjanids in the tropical Indo-West Pacific. Most of the putative species within this complex are yet to be described. Comparison of identical numbers of matched samples of fishes from French Polynesia, Heron Island (southern GBR) and Lizard Island (northern GBR) revealed 1, 4 and 10 species of Transversotrema respectively suggesting that the French Polynesian fauna is depauperate for this family. In addition to those species apparently missing from suitable hosts in French Polynesia, several species from further west infect fishes (especially Nemipteridae) that are themselves absent from French Polynesia. This dramatic east-west decline in richness contrasts strongly with what is known for monogeneans, which appear to maintain their richness over the same scale, and is more precipitate than is known for other groups of trematodes. The decline might be explained in part by the absence of the as yet unknown first intermediate hosts in French Polynesia. However, we predict that it is explained by other life cycle traits. We hypothesise that the characters of large short-lived cercariae, short-lived miracidia, the absence in the life-cycle of second intermediate hosts that are capable of transporting the species, and definitive and first intermediate hosts that have limited vagility combine to give marine Transversotrematidae limited dispersal capacity and a propensity for localised speciation.

  7. Infrapopulations of Gyliauchen volubilis Nagaty, 1956 (Trematoda: Gyliauchenidae) in the rabbitfish Siganus rivulatus (Teleostei: Siganidae) from the Saudi coast of the Red Sea.

    PubMed

    Al-Jahdali, M O

    2012-08-01

    In hermaphroditic helminth parasites, infrapopulation size or mating group size mostly affects some processes acting within the infrapopulation. Here, 30 natural infrapopulations (12-154 individuals) of the intestinal trematode Gyliauchen volubilis Nagaty, 1956 from the fish Siganus rivulatus consisting of newly excysted juveniles, immature and mature worms were found distributed in a well-defined fundamental niche (anterior 40% of the intestine). In small infrapopulations, all stages of the parasite were alive. In larger infrapopulations, differential mortality was only and consistently observed among newly excysted juveniles, and gradually increased to include most or all juveniles in the largest infrapopulations. Among mature worms, the mean worm length seemed unaffected by the infrapopulation size. However, the ratio mean testis size-mean ovary size, a reliable indicator of resource allocation to the male function and of opportunities for cross fertilization, significantly increased with mating group size. In small infrapopulations, all stages of the parasite were scattered along the niche, and never seen in mating pairs (possibly reproduced by self-fertilization). In larger infrapopulations, newly excysted juveniles and immature worms were scattered along the anterior two thirds of the niche, while mature worms were constantly found aggregated in its posterior third (narrow microhabitat), where some were arranged in mating pairs. The probability of mating reciprocally or unilaterally was dependent on body size. The mean number of uterine eggs per worm significantly decreased and their mean sizes significantly increased with mating group size. The results are statistically significant and suggest that infrapopulation self-regulation is greatly associated with its size.

  8. Physa marmorata (Mollusca: Physidae) as a natural intermediate host of Trichobilharzia (Trematoda: Schistosomatidae), a potential causative agent of avian cercarial dermatitis in Brazil.

    PubMed

    Pinto, Hudson A; Brant, Sara V; de Melo, Alan L

    2014-10-01

    Species of Trichobilharzia are the main etiological agents of cercarial dermatitis in humans, which is considered a re-emerging disease. Despite the diversity and global distribution of species of this genus, studies of Trichobilharzia are scarce in South America. The goal of our investigations is better understand the diversity, distribution and life cycle of avian schistosomes and their likely role in causing dermatitis in Brazil. As part of this effort, cercariae found in naturally infected Physa marmorata were identified by morphological and molecular (mitochondrial cox1, nuclear ITS1 and 28S gene regions) methods as Trichobilharzia sp. These cercariae are similar morphologically to T. jequitibaensis described previously from Brazil and similar genetically to the North American physid transmitted species T. querquedulae and T. physellae. This is the first report of a potential agent of cercarial dermatitis from naturally infected snails from Brazil and first molecular characterization of a South American species of Trichobilharzia. A discussion follows concerning the potential role of this species has in outbreaks of dermatitis in Brazil.

  9. Where are the South American freshwater turtle blood flukes (Trematoda: Spirorchiidae)? The first morphological and molecular analysis of spirorchiid cercariae from freshwater snails in Brazil.

    PubMed

    Pinto, Hudson Alves; de Melo, Alan Lane; Brant, Sara Vanessa

    2015-12-01

    Trematodes belonging to the family Spirorchiidae are blood parasites mainly of turtles with a worldwide distribution. These flukes were recently reported in some marine turtles from South America, where the occurrence of spirorchiids in freshwater definitive and intermediate hosts is so far unknown. In the present study, three morphotypes of brevifurcate apharyngeate distome cercariae found in freshwater molluscs from an urban reservoir in Brazil were used for morphological and molecular (nuclear 28S rDNA) evaluation. Two morphotypes of cercariae, probably congeneric species, were found in 12/17,465 specimens of Biomphalaria spp. and differ from each other by body size and sequences (0.1%). They present morphology similar to North American freshwater spirorchiids (Spirorchis spp.), however surprisingly molecular data reveals that these lineages are more closely related to marine spirorchiids. A third species found in 2/777 Pomacea sp. differs morphologically from all previously described spirorchiid cercariae and genetically from spirorchiids with available sequences (16-19%), grouping in the phylogenetic tree with freshwater North American species. This is the first report of freshwater spirorchiids in South America and the first molecular confirmation of the involvement of a caenogastropod in the life cycle of spirorchiids. PMID:26253761

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

    PubMed

    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. PMID:27216231

  11. Evaluation of the mitochondrial system in the gonad-digestive gland complex of Biomphalaria glabrata (Mollusca, Gastropoda) after infection by Echinostoma paraensei (Trematoda, Echinostomatidae).

    PubMed

    Tunholi, Victor Menezes; Tunholi-Alves, Vinícius Menezes; Santos, Anderson Teixeira; Garcia, Juberlan da Silva; Maldonado, Arnaldo; da-Silva, Wagner Seixas; Rodrigues, Maria de Lurdes de Azevedo; Pinheiro, Jairo

    2016-05-01

    The effect of infection by Echinostoma paraensei on the mitochondrial physiology of Biomphalaria glabrata was investigated after exposure to 50 miracidia. The snails were dissected one, two, three and four weeks after infection for collection and mechanical permeabilization of the gonad-digestive gland (DGG) complex. The results obtained indicate that prepatent infection by this echinostomatid fluke significantly suppresses the phosphorylation state (respiratory state 3) and basal oxygen consumption of B. glabrata, demonstrating that the infection reduces the ability of the intermediate host to carry out aerobic oxidative reactions. Additionally, relevant variations related to the uncoupled mitochondrial (state 3u) of B. glabrata infected by E. paraensei were observed. Four weeks after exposure, a significant reduction in mitochondrial oxygen consumption after addition of ADP (3.68±0.26pmol O2/mg proteins) was observed in the infected snails in comparison with the respective control group (5.14±0.25). In the uncoupled state, the infected snails consumed about 62% less oxygen than the infected snails (7.87±0.84pmol O2/mg proteins) in the same period. These results demonstrate a reduction in oxidative decarboxylation rate of the tricarboxylic acid cycle and faster anaerobic degradation of carbohydrates in the infected snails. The possible mechanisms that explain this new metabolic condition in the infected organisms are discussed.

  12. Detoxifying and anti-oxidant enzymes of Fasciola gigantica worms under triclabendazole sulphoxide (TCBZ-SX): an in vitro study.

    PubMed

    Shehab, Amel Y; Ebeid, Samia M; El-Samak, Mohamed Y; Hussein, Neveen M

    2009-04-01

    Triclabendazole (TCBZ) is an efficient fasciolicide that affects both juvenile and adult worms. After oral administration it is rapidly metabolized to TCBZ sulphoxide and sulphone that were found responsible for its fasciolicidal activity. Parasite defense mechanisms include detoxifying and anti-oxidant enzymes that would suppress its oxidative killing. The present work aimed at evaluating these enzymes under TCBZ-SX. Thirty juvenile and 30 adult F. gigantica worms collected from the liver parenchyma and bile ducts formed the subject of the study. Levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), and glutathione (GSH) were measured in juvenile and adult worms incubated, without and with 15 and 50 microg/ml TCBZ-SX for 18 hours at 37 degrees C. Results demonstrated a significant decrease in SOD activity and an increase in GST level in both juvenile and adult worms after incubation in the two concentrations. The remaining enzymes and GSH revealed variable levels.

  13. Diagnosis and economic consequences of triclabendazole resistance in Fasciola hepatica in a sheep flock in south-east Scotland.

    PubMed

    Sargison, N D; Scott, P R

    2011-02-12

    Over the past decade, definite changes have been recorded in the regional prevalence, seasonality and severity of fasciolosis in the UK, related to increased rainfall, or localised flooding, prompting debate about the deleterious effects of climate change. As a consequence, effective management of fasciolosis has become problematic in areas where fluke traditionally exists, leading to serious loss of production in sheep and cattle. Meanwhile, in eastern districts, there have been unexpected outbreaks of disease, resulting in production losses and concerns about welfare. This case report describes the economic consequences of fasciolosis in a commercial sheep flock in south-east Scotland. The diagnosis and consequences of triclabendazole resistance are discussed, in the context of developing economically sustainable control strategies. PMID:21493511

  14. [Influence of vaccination of calves with recombinant cysteine proteinase of Fasciola hepatica on development and infectivity of miracidia].

    PubMed

    Dabrowska, Martyna; Kaliniak, Marcin; Wedrychowicz, Halina

    2006-01-01

    The aim of the study was to evaluate an influence of vaccination of the final host on F. hepatica development in intermediate hosts. Fluke eggs were isolated from the biliary tracts of calves vaccinated orally with recombinant cysteine proteinase of F. hepatica after the challenge infection and from control calves which received the infection only. To asses the effect of the vaccine on egg "hatch rate" the eggs were transferred to the Petri dishes with distilled water and incubated at 25 degrees C for 16-19 days. They were subsequently exposed to light for about 2 h, at a temperature of 27 +/- 1 degrees C, to stimulate sprouting of the miracidia and asses the egg hatchability. In order to evaluate infectivity and pathogenicity of the miracidia, single miracidium infections of Lymnea truncatula by F. hepatica were carried out under laboratory conditions using 4-mm-high snails. The prevalence of snail infections with F. hepatica was calculated using the ratio between the number of cercariae-shedding snails in each group and that of surviving snails. It appeared that the eggs isolated from immunized calves demonstrated significantly lower hatchability than the eggs isolated from non-vaccinated control hosts. Also, the proportion of infected snails as well as their mortality were lower after exposition to miracidia originating from vaccinated calves. It is suggested that effectors of the immune response in vaccinated calves inhibited in part biological activity of cysteine proteinases of the fluke which are known to be involved in egg shell formation, penetration of host's tissues and worm feeding. PMID:17432623

  15. Fasciola hepatica vaccine: We may not be there yet but we’re on the right road

    PubMed Central

    Molina-Hernández, Verónica; Mulcahy, Grace; Pérez, Jose; Martínez-Moreno, Álvaro; Donnelly, Sheila; O’Neill, Sandra M.; Dalton, John P.; Cwiklinski, Krystyna

    2015-01-01

    Major advances have been made in identifying potential vaccine molecules for the control of fasciolosis in livestock but we have yet to reach the level of efficacy required for commercialisation. The pathogenesis of fasciolosis is associated with liver damage that is inflicted by migrating and feeding immature flukes as well as host inflammatory immune responses to parasite-secreted molecules and tissue damage alarm signals. Immune suppression/modulation by the parasites prevents the development of protective immune responses as evidenced by the lack of immunity observed in naturally and experimentally infected animals. In our opinion, future efforts need to focus on understanding how parasites invade and penetrate the tissues of their hosts and how they potentiate and control the ensuing immune responses, particularly in the first days of infection. Emerging ‘omics’ data employed in an unbiased approach are helping us understand liver fluke biology and, in parallel with new immunological data, to identify molecules that are essential to parasite development and accessible to vaccine-induced immune responses. PMID:25657086

  16. [Parasites of the Osprey (Pandion haliaetus) in northwestern Russia].

    PubMed

    Lebedeva, D I; Iakovleva, G A; Artem'ev, A V

    2013-01-01

    New data on Ospey parasites in Karelian Republic are given. One specimen was investigated. Two parasite species--Nematostrigea serpens and Diplostomum pseudospathaceum were found. Trematoda D. pseudospathaceum was recorded in Osprey parasite fauna for the first time. PMID:24455873

  17. Phylogenetic position of Magnivitellinum Kloss, 1966 and Perezitrema Baruš & Moravec, 1967 (Trematoda: Plagiorchioidea: Macroderoididae) inferred from partial 28S rDNA sequences, with the establishment of Alloglossidiidae n. fam.

    PubMed

    Hernández-Mena, David Iván; Mendoza-Garfias, Berenit; Ornelas-García, Claudia Patricia; Pérez-Ponce de León, Gerardo

    2016-07-01

    The systematic position of two genera of Macroderoididae McMullen, 1937, Perezitrema Baruš & Moravec, 1967 and Magnivitellinum Kloss, 1966 is reviewed based on a phylogenetic analysis of the interrelationships of 15 species of the family allocated into six genera, along with 44 species of plagiorchioid trematodes, using partial sequences of the 28S rRNA gene. Sequences were analysed through parsimony, maximum likelihood and Bayesian inference. The obtained topologies show Perezitrema as the sister taxon of three species of Macroderoides Pearse, 1924; the latter genus appears to be paraphyletic since another three species are not included in this group. Instead, Magnivitellinum was placed as the sister taxon of Alloglossidium Simer, 1929. These relationships are well supported by high bootstrap and posterior probability values. The resulting trees demonstrate that the family Macroderoididae, as currently conceived in taxonomic treatments, is not monophyletic. Magnivitellinum simplex Kloss, 1966 and Alloglossidium spp. were nested as sister taxa of members of the family Leptophallidae Dayal, 1938, whereas Perezitrema bychowskii Baruš & Moravec, 1967 and species of Macroderoides and Paramacroderoides Venard, 1941 were grouped with Auridistomum chelydrae (Stafford, 1900), a monotypic member of Auridistomidae Stunkard, 1924. Based on our results, a new family, Alloglossidiidae n. fam. was established to accommodate the genera Magnivitellinum and Alloglossidium. PMID:27307166

  18. INFLUENCE OF RIBEIROIA ONDATRAE (TREMATODA: DIGENEA) INFECTION ON LIMB DEVELOPMENT AND SURVIVAL OF NORTHERN LEOPARD FROGS (RANA PIPIENS): EFFECTS OF HOST STAGE AND PARASITE-EXPOSURE LEVEL. (R825867)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  19. The Apple Snail Pomacea maculata (Caenogastropoda: Ampullariidae) as the intermediate host of Stomylotrema gratiosus (Trematoda: Stomylotrematidae) in Brazil: the first report of a mollusc host of a Stomylotrematid Trematode.

    PubMed

    Pinto, Hudson A; Cantanhede, Selma Patrícia D; Thiengo, Silvana C; de Melo, Alan L; Fernandez, Monica A

    2015-04-01

    Trematodes belonging to the family Stomylotrematidae are intestinal parasites of birds. Despite the worldwide distribution and diversity of host species, the first intermediate host remains unknown. For a survey of parasites of Pomacea maculata , snails were collected from the municipality of São Vicente Férrer, state of Maranhão, northeastern Brazil. In the present study, the xiphidiocercariae shed from these snails were used in the experimental infection of the water bug Belostoma plebejum. The insect mortality was observed 30 days post-infection, and the metacercariae recovered in the body cavity of B. plebejum were identified as Stomylotrema gratiosus. This is the first report of an ampullariid snail as intermediate host of stomylotrematid trematodes.

  20. Parallopharynxs pp. (Trematoda: Digenea: Plagiorchioidea) in iguanian lizards from the Area de Conservación Guanacaste, Guanacaste, Costa Rica, including Parallopharynx matternae n. sp. in Chaetodon basiliscus (Squamata: Iguania: Corytophanidae).

    PubMed

    Zamparo, David; Brooks, Daniel R; Tkach, Vasyl; Rodriguez, Beatriz

    2004-04-01

    We report 3 species of the digenean genus Parallopharynx, 1 previously undescribed, from the Area de Conservación Guanacaste (ACG) in northwestern Costa Rica. Parallopharynx gonzalezi, which was originally described in Basiliscus sp. and Ctenosoura similis from central Costa Rica, inhabits C. quinquecarinata; P. jonesi, originally described in Anolis lionotus (syn. Norops oxylophus) from Nicaragua, inhabits N. oxylophus, N. biporcatus, and Basiliscus basiliscus; and the new species, which inhabits B. basiliscus. Parallopharynx matternae n. sp. differs from all other members of the genus by having a metraterm extending posteriad from the genital pore to the posterior margin of the ventral sucker, whereas in P. arctus and P. gonzalezi, the metraterm never surpasses the midlevel of the ventral sucker and in P. jonesi it never passes the anterior margin of the ventral sucker, and by having an oral sucker that does not exceed 150 microm in diameter with a subsequent greater oral-ventral sucker width ratio ranging from 1:0.88-1.12 (averaging 1:1), whereas values range from 1:0.71-0.83 for P. gonzalezi and P. arctus, and from 1:0.59-0.68 for P. jonesi. Parallopharynx spp. possesses Y-shaped excretory vesicles with a long central stem and short arms bifurcating immediately posterior to the ovary; similar to those found in members of the Telorchiidae. Additional similarities in the relative positions of the gonads and the structure of the cirrus sac and metraterm indicate a close relationship between Parallopharynx and members of the Telorchiidae.

  1. Wallinia chavarriae n. sp. (Trematoda: Macroderoididae) in Astyanax aeneus (Gunther, 1860) and Bryconamericus scleroparius (Regan, 1908) (Osteichthyes: Characidae) from the Area de Conservación Guanacaste, Costa Rica.

    PubMed

    Choudhury, Anindo; Daverdin, Rita Hartvigsen; Brooks, Daniel R

    2002-02-01

    Wallinia chavarriae n. sp. is described from the small-bodied characids Astyanax aeneus and Bryconamericus scleroparius in the Area de Conservación Guanacaste, northwestern Costa Rica. The species differs from W. valenciae in possessing an acetabulum that is smaller than the oral sucker and vitelline follicles that are ovoid or rounded rather than elongate and tubular. Detailed comparison between these 2 species is handicapped by the less than satisfactory condition of the type and only museum specimen of W. valenciae. Wallinia chavarriae and W. valenciae belong to a subfamily of trematodes, Walliniinae, that arguably includes Creptotrematina spp., Magnivitellum simplex, and possibly Margotrema. The morphology of walliniines suggests that they are macroderoidids, but a clearer understanding of their classification could be gained from their larval morphology or from molecular systematic studies. The host associations of a monophyletic Walliniinae would indicate diversification within 2 groups of freshwater fishes: the neotropical characids for species of Wallinia, Creptotrematina, and Magnivitellum and the endemic central Mexican goodeids for those of Margotrema. The biogeography and host associations of these parasites provide a system for studies of potential host switching and vicariance, involving the middle-American and neotropical regions.

  2. Studies on the ultrastructure and histochemistry of the lymph system in three species of amphistome (Trematoda: Digenea) Gigantocotyle explanatum, Gastrothylax crumenifer and Srivastavaia indica from the Indian water buffalo Bubalus bubalis.

    PubMed

    Dunn, T S; Nizami, W A; Hanna, R E

    1985-03-01

    The lymph system of three amphistome parasites from buffaloes, Gigantocotyle explanatum, Gastrothylax crumenifer and Srivastavaia indica was studied using light microscope histochemistry and electron microscopy. In each case the system comprised a single pair of main longitudinal vessels which gave rise to numerous sub-dividing lateral branches. Although the finer lymph channels associated with most internal systems, they did not penetrate the basement membrane of any organ. The lymph vessels were delimited by a unit membrane and separated from adjacent cells by interstitial material. The lymph fluid consisted of an amorphous proteinaceous, lipid-rich matrix, containing naked nuclei and granules of various sizes. Complexes of endoplasmic reticulum were frequently associated with the nuclei. No distinct Golgi bodies or mitochondria were evident. The granules noted throughout the lymph morphologically resembled autophagosomes and lysosomes. Autophagy within the lymph system presumably mobilizes amino acids for subsequent transport to tissues undergoing active protein synthesis. The lymph channels displayed an intimate relationship with the general parenchyma. In particular, numerous protrusions of lymph occurred into the cytoplasm of certain specialized parenchymal cells surrounding the pharynx. Within these 'juxtapharyngeal' cells autophagic degradation of sequestered lymph cytoplasm apparently occurred. In the three species of amphistome studied, the lymph system appears to function in storage and mobilization of amino acids and possibly lipids. It may also serve to distribute other small molecules throughout the body. The detection of haemoglobin in the lymph system of G. crumenifer and S. indica, but not in Gigantocotyle explanatum, suggests a further role in oxygen storage and transport.

  3. Herd evaluation by enzyme-linked immunosorbent assay for the determination of Fasciola hepatica infection in sheep and cattle from the Altiplano of Bolivia.

    PubMed

    Hillyer, G V; Soler de Galanes, M; Buchón, P; Bjorland, J

    1996-02-01

    A study was designed to determine by ELISA the seroprevalence of fasciolosis both in sheep (29 herds totaling 184 sheep), in samples collected in 1988, and in cattle (41 herds totaling 299 animals, samples collected in 1988; 34 herds totaling 147 animals, samples collected in 1989) in the same area of Corapata in which a seroprevalence survey had been done in humans. The results show high seropositivity in sheep (89%) and lower seropositivity in cattle (58% in 1988, and 57% in 1989). The seroprevalence in cattle in 1988 was essentially identical to that detected in 1989. Faecal examinations were also done in the 1988 sheep and 1989 cattle. Results of the study showed that of the 184 sheep examined, 22 were positive for F. hepatica eggs, while 163 were positive by serology. All of the 22 sheep which were positive parasitologically were also positive serologically for a sensitivity of 100%. On the other hand, of 147 cattle tested, 38 were positive parasitologically while 84 were positive serologically. Of the 38 positives for F. hepatica eggs, 31 were positive by serology (sensitivity 82%).

  4. Fasciola hepatica - monitoring the milky way? The use of tank milk for liver fluke monitoring in dairy herds as base for treatment strategies.

    PubMed

    Duscher, Ruth; Duscher, Georg; Hofer, Johannes; Tichy, Alexander; Prosl, Heinrich; Joachim, Anja

    2011-06-10

    In this study 595 lactating cows originating from 31 carinthian farms were investigated in accordance of liver fluke infection using individual and tank milk as well as individual blood and faecal samples. Two commercial ELISAs were used to test the milk and blood serum, and the results were compared with coproscopy and a commercial copro-antigen ELISA. Receiver operating characteristics (ROC) and two-graph operating characteristics (TG ROC) of tank milk results were conducted based on the individual milk to determine the minimum reliable in-herd antibody prevalence for the predominant condition in the investigation area. In 17.8% of the examined individuals located in 64.5% of the farms eggs were detected by coproscopy. The copro-antigen ELISA delivered 13.4% positive individuals from 54.8% of the farms. The milk ELISAs showed 42.7% (Euroclone) and 44.2% (Pourquier) positive cows on 90.3% of the farms. The blood samples were positive in 43% (Euroclone) and 45.2% (Pourquier) of the individuals from 90.3% to 96.8% of the herds, respectively. Based on the milk and the blood an average in-herd prevalence of 30-45% can be assumed. The serum and milk samples delivered correlating results with kappa values between 0.94 and 0.97, whereas the coproscopy and copro-antigen ELISA did not correlate well with the ELISA results. The two different ELISA tests highly correlated on individual and on herd level. Both showed a reliable minimum in-herd prevalence of ∼20%, meaning that one fifth of the individuals in a herd have to be positive to obtain a positive bulk tank milk result. In the investigated area a higher in-herd prevalence is expected, therefore the tank milk is useful as a monitoring tool and can be used as a basis for intervention strategies.

  5. The occurrence of the snail Lymnaea columella on grazing areas in New South Wales and studies on its susceptibility to Fasciola hepatica.

    PubMed

    Boray, J C; Fraser, G C; Williams, J D; Wilson, J M

    1985-01-01

    Field surveys were carried out in the Lismore and Casino area for the presence of fresh water snails potentially responsible for the transmission of trematodes in ruminants. Although the North American snail, Lymnaea columella has previously only been reported from metropolitan areas, large populations of the snail were found east of Lismore. Natural infection with F. hepatica was detected in some of the snails and laboratory studies showed that this snail species was highly susceptible to infection with miracidia of F. hepatica and produced viable metacercariae. Three species of planorbid snails, Helicorbis australiensis, Pygmanisus pelorius and Gyraulus gilberti were also found. The first 2 planorbid species were infected with paramphistomid cercariae.

  6. Evaluation of the Performance of Five Diagnostic Tests for Fasciola hepatica Infection in Naturally Infected Cattle Using a Bayesian No Gold Standard Approach.

    PubMed

    Mazeri, Stella; Sargison, Neil; Kelly, Robert F; Bronsvoort, Barend M deC; Handel, Ian

    2016-01-01

    The clinical and economic importance of fasciolosis has been recognised for centuries, yet diagnostic tests available for cattle are far from perfect. Test evaluation has mainly been carried out using gold standard approaches or under experimental settings, the limitations of which are well known. In this study, a Bayesian no gold standard approach was used to estimate the diagnostic sensitivity and specificity of five tests for fasciolosis in cattle. These included detailed liver necropsy including gall bladder egg count, faecal egg counting, a commercially available copro-antigen ELISA, an in-house serum excretory/secretory antibody ELISA and routine abattoir liver inspection. In total 619 cattle slaughtered at one of Scotland's biggest abattoirs were sampled, during three sampling periods spanning summer 2013, winter 2014 and autumn 2014. Test sensitivities and specificities were estimated using an extension of the Hui Walter no gold standard model, where estimates were allowed to vary between seasons if tests were a priori believed to perform differently for any reason. The results of this analysis provide novel information on the performance of these tests in a naturally infected cattle population and at different times of the year where different levels of acute or chronic infection are expected. Accurate estimates of sensitivity and specificity will allow for routine abattoir liver inspection to be used as a tool for monitoring the epidemiology of F. hepatica as well as evaluating herd health planning. Furthermore, the results provide evidence to suggest that the copro-antigen ELISA does not cross-react with Calicophoron daubneyi rumen fluke parasites, while the serum antibody ELISA does. PMID:27564546

  7. Evaluation of the Performance of Five Diagnostic Tests for Fasciola hepatica Infection in Naturally Infected Cattle Using a Bayesian No Gold Standard Approach

    PubMed Central

    Sargison, Neil; Kelly, Robert F.; Bronsvoort, Barend M. deC.; Handel, Ian

    2016-01-01

    The clinical and economic importance of fasciolosis has been recognised for centuries, yet diagnostic tests available for cattle are far from perfect. Test evaluation has mainly been carried out using gold standard approaches or under experimental settings, the limitations of which are well known. In this study, a Bayesian no gold standard approach was used to estimate the diagnostic sensitivity and specificity of five tests for fasciolosis in cattle. These included detailed liver necropsy including gall bladder egg count, faecal egg counting, a commercially available copro-antigen ELISA, an in-house serum excretory/secretory antibody ELISA and routine abattoir liver inspection. In total 619 cattle slaughtered at one of Scotland’s biggest abattoirs were sampled, during three sampling periods spanning summer 2013, winter 2014 and autumn 2014. Test sensitivities and specificities were estimated using an extension of the Hui Walter no gold standard model, where estimates were allowed to vary between seasons if tests were a priori believed to perform differently for any reason. The results of this analysis provide novel information on the performance of these tests in a naturally infected cattle population and at different times of the year where different levels of acute or chronic infection are expected. Accurate estimates of sensitivity and specificity will allow for routine abattoir liver inspection to be used as a tool for monitoring the epidemiology of F. hepatica as well as evaluating herd health planning. Furthermore, the results provide evidence to suggest that the copro-antigen ELISA does not cross-react with Calicophoron daubneyi rumen fluke parasites, while the serum antibody ELISA does. PMID:27564546

  8. [Epidemiologic data on distomiasis due to Fasciola gigantica Cobbold on the island of Madagascar. Infestation of cattle between 1975 and 1981].

    PubMed

    Rakotondravao; Rondelaud, D

    1985-01-01

    Stools examinations of zebus were made in Madagascar between 1975 and 1981 for determining the importance of Fascioliasis gigantica in the six provinces of the island during this time. High percentages of F. gigantica eggs were found in the stools of the different infected zebus-herds of five provinces. No fasciolid infection was noted in 1979 in the cattle of the province of Diego-Suarez. Differences were noted between the infection rate obtained by fecal examination and the rate indicated in the veterinary reports. The significance of these results is discussed.

  9. Clinostomum marginatum in steelhead trout (Salmo gairdneri) and cutthroat trout (Salmo clarki) in a western Washington lake

    USGS Publications Warehouse

    Uzmann, J.R.; Douglas, J.

    1966-01-01

    Clinostomum marginatum (Trematoda: Clinostomatidae), the yellow grub parasite, was recorded in epizootic proportions from Lynch Lake, King County, Washington, in 1961 and 1962. The parasite larvae occurred principally in steelhead trout (Salmo gairdneri); cutthroat trout (S. clarki) were infected to a relatively minor degree. Fish and snail host populations were destroyed by rotenone and copper sulfate treatments.

  10. NEW DATA ON BIRD HELMINTHS IN MONGOLIA.

    PubMed

    Lebedeva, D I; Chantuu, K

    2015-01-01

    For the first time the data on helminths in piscivorous birds (the great cormorant and the Mongolian gull) in Mongolia were obtained. Surveys yielded 11 species (Cestoda--2, Trematoda--6, Nematoda--3). The cormorant hosted 5 helminth species, the herring gull--6 species. PMID:26827489

  11. Urotocus rossittensis (Trematoda: Digenea: Leucochloriidae) in the scarlet-rumped tanager, Ramphocelus passerinii, and common bush tanager, Chlorospingus ophthalmicus (Passeriformes: Thraupidae), from the Area de Conservación Guanacaste, Costa Rica, with taxonomic revision of the genus and revised key to the leuchochloridiid-like Brachylaimoidea.

    PubMed

    Zamparo, David; Brooks, Daniel R

    2006-06-01

    Urotocus rossittensis occurs in the bursa Fabricii of the scarlet-rumped tanager, Ramphocelus passerinii, and the common bush tanager, Chlorospingus ophthalmicus, from the Area de Conservación Guanacaste, Costa Rica. Morphological examination of type material of U. fusiformis and U. kenyensis and Costa Rican specimens suggests that U. fusiformis and U. kenyensis are indistinguishable from U. rossittensis. Confirmed accounts of Urotocus spp. refer to a single adult morphotype whose geographic distribution includes the Palearctic, Africa, Nearctic, and northern Neotropics.

  12. Whallwachsia illuminata n. gen., n. sp. (Trematoda: Digenea: Plagiorchiformes: Prosthogonimidae) in the steely-vented hummingbird Amazilia saucerrottei (Aves: Apodiformes: Trochilidae) and the yellow-olive flycatcher Tolmomyias sulphurescens (Aves: Passeriformes: Tyraninidae) from the Area de Conservación Guanacaste, Guanacaste, Costa Rica.

    PubMed

    Zamparo, David; Brooks, Daniel R; Causey, Douglas

    2003-08-01

    A new species of digenean found in the intestines of the steely-vented hummingbird Amazilia saucerrottei and the yellow-olive flycatcher Tolmomyias sulphurescens from the Area de Conservación Guanacaste, Guanacaste, Costa Rica, resembles members of the Prosthogonimidae in having a highly lobate ovary; an elongate cirrus sac containing the cirrus, pars prostatica, and internal seminal vesicle; no external seminal vesicle; 2 fields of extracecal vitelline follicles restricted to the area between the intestinal bifurcation and testes; and uterine loops occupying all available space in the hind body. The new species differs from all other members of the family in having genital pores opening laterally to the cecum, immediately anterior to the acetabular level, and markedly oblique rather than symmetrical testes. Consequently, we propose the new genus Whallwachsia for the species. Preliminary phylogenetic assessment suggests that the species is the sister group of all other prosthogonimids.

  13. Prevalence of fasciolosis and dicrocoeliosis in slaughtered sheep and goats in Amol Abattoir, Mazandaran, northern Iran

    PubMed Central

    Khanjari, Ali; Bahonar, Alireza; Fallah, Sepideh; Bagheri, Mahboube; Alizadeh, Abbas; fallah, Marjan; Khanjari, Zahra

    2014-01-01

    Objective The liver flukes, Fasciola spp. and Dicrocoelium dendriticum, infect ruminants and other mammalian extensively and cause major diseases of livestock that produce considerable economic losses. Methods A survey of 2391 sheep and goats slaughtered at an abattoir in Amol region, northern Iran was used to determine the prevalence of the liver flukes infection based on season, sex and specie of the animals. Results The results revealed that the prevalence rate of Fasciola spp. and Dicrocoelium dendriticum was 6.6% and 4.3% respectively. Dicrocoeliosis was more dominant in female animals (7.1%) whereas there was no sex-related difference in the prevalence of Fasciola spp. in male and female animals. Furthermore, Fasciolosis was significantly more prevalent than dicrocoeliosis in both sheep and goats. The Seasonal prevalence of Fasciola spp. was highest (P<0.005) during spring (8.3%) followed in order by autumn (8.1%), winter (5.9%) and summer (4.0%) but Dicrocoeliosis did not follow any seasonal pattern. Conclusions According to this study, it can be concluded that Amol is regarded as an endemic region for Fasciola spp and D. dandriticum infection. Moreover, Fasciola spp. is the most widespread liver fluke found in sheep and goats which is more dominant in sheep than goats.

  14. [Helminth fauna of amphibians (Vertebrata: Amphibia) in the Republic of Belarus].

    PubMed

    Shimalov, V V

    2009-01-01

    Historical review of the investigations of helminth fauna in amphibians from Belarus is presented. In 12 amphibian species examined by different authors 46 helminth species were found, including 29 Trematoda, 13 Nematoda, 1 Monogenea, 2 Cestoda, and 1 Acanthocephala. Original data on helminths parasitizing Amphibia in Byelorussian Polesie, by the results of long-term investigations in 1986-2004 are given. Distribution of 40 helminth species by hosts and respective infestation rates are reported.

  15. [Helminthofauna of wild canids in Azerbaijan and ways of its formation].

    PubMed

    Fataliev, G G

    2011-01-01

    The complete list of helminthes parasitizing canids in Azerbaijan is given; ways of formation of the canids' helminthofauna in Azerbaijan are reconstructed. As a result of our study, 42 helminth species were recorded; 25 of them parasitize jackals, 16 parasitize wolfs, and 39 species were found in foxes. The helminthofauna includes 5 species of Trematoda, 14 species of Cestoda, 1 species of Acanthocephala, and 22 species of Nematoda. By the life cycle, 32 species belong to biohelminthes and 10 species are geohelminthes.

  16. Parasites in pet reptiles

    PubMed Central

    2011-01-01

    Exotic reptiles originating from the wild can be carriers of many different pathogens and some of them can infect humans. Reptiles imported into Slovenia from 2000 to 2005, specimens of native species taken from the wild and captive bred species were investigated. A total of 949 reptiles (55 snakes, 331 lizards and 563 turtles), belonging to 68 different species, were examined for the presence of endoparasites and ectoparasites. Twelve different groups (Nematoda (5), Trematoda (1), Acanthocephala (1), Pentastomida (1) and Protozoa (4)) of endoparasites were determined in 26 (47.3%) of 55 examined snakes. In snakes two different species of ectoparasites were also found. Among the tested lizards eighteen different groups (Nematoda (8), Cestoda (1), Trematoda (1), Acanthocephala (1), Pentastomida (1) and Protozoa (6)) of endoparasites in 252 (76.1%) of 331 examined animals were found. One Trombiculid ectoparasite was determined. In 563 of examined turtles eight different groups (Nematoda (4), Cestoda (1), Trematoda (1) and Protozoa (2)) of endoparasites were determined in 498 (88.5%) animals. In examined turtles three different species of ectoparasites were seen. The established prevalence of various parasites in reptiles used as pet animals indicates the need for examination on specific pathogens prior to introduction to owners. PMID:21624124

  17. Screening for Zoonotic Fascioliasis in Slaughtered Large Ruminants in Abattoirs in Perak, Malaysia

    PubMed Central

    Zainalabidin, Fazly Ann; Azmi, Muhamad Syamsul Naim Noor; Bakri, Wan Normaziah Wan Omar; Sathaya, Geethamalar; Ismail, Mohd Iswadi

    2015-01-01

    Fascioliasis, or trematode infestation, is an important disease caused by Fasciola hepatica and Fasciola gigantica. Both species are hepatic parasites that affect humans. We have examined the zoonotic aspects of fascioliasis. A total of 80 fresh liver samples were collected from 67 Kedah-Kelantan crossbred cattle and 13 Murrah buffalo at 4 local abattoirs in Perak, Malaysia. The samples were examined macroscopically to detect the presence of Fasciola spp. The results show 7.50% (6 of 80) of the animals were diagnosed with fascioliasis. Overall, 7.46% (5 of 67) and 7.69% (1 of 13) of cattle and buffalo samples were positive, respectively. There were only F. gigantica species identified in the samples. Our findings suggest that precautions should be taken because the disease has a zoonotic impact on public health. PMID:26868715

  18. High prevalence of large trematode eggs in schoolchildren in Cambodia.

    PubMed

    Bless, Philipp J; Schär, Fabian; Khieu, Virak; Kramme, Stefanie; Muth, Sinuon; Marti, Hanspeter; Odermatt, Peter

    2015-01-01

    Large trematode eggs (LTE) resembling Fasciola spp. eggs were reportedly found in the stools of schoolchildren in Kandal province, Cambodia. This study aimed to assess the prevalence of LTE in the stools of children attending the affected school, identify potential risk factors for infection and ascertain the trematode species. We performed a cross-sectional study involving an in-depth questionnaire administered to schoolchildren at the affected school, and examined cattle droppings in the surrounding area and the livers of slaughtered cattle. Three stool samples were examined per child, using Kato-Katz and formalin-ether concentration techniques. In addition, blood serum enzyme-linked immunosorbent assay (ELISA) and coprological polymerase chain reaction (PCR) was conducted for species clarification. Cattle droppings were examined by cup sedimentation and coprological ELISA. LTE were observed in the stools of 106 schoolchildren (46.5%). Two blood serum samples from schoolchildren were positive for Fasciola hepatica in a first ELISA but were negative in a confirmation immunofluorescence antibody test. Out of 221 PCR samples, only one tested positive for Fasciola spp. and none for Fasciolopsis buski. The consumption of raw aquatic plants (odds ratio (OR)=3.3) and fermented fish sauce (OR=2.1) were significantly associated with LTE in the stool. Fasciola spp. flukes were observed in 18.3% of 191 cattle livers. The prevalence of fascioliasis in cattle droppings was 88.8%. The low prevalence of schoolchildren that tested positive for Fasciola spp. with specific molecular diagnostics and who had no diagnostic evidence of F. buski strongly indicates that the majority of microscopically observed LTE are from Echinostoma spp. Fasciola spp. transmission from cattle to human is possible and public health services need to be alerted accordingly.

  19. Progress in development of liver fluke vaccines.

    PubMed

    Spithill, T W; Dalton, J P

    1998-12-01

    Infection of ruminants by Fasciola spp continues to cause large economic losses worldwide. Recent results from several laboratories have demonstrated that animals can be significantly protected against infection by vaccination with defined Fasciola antigens. Apart from reducing fluke burdens, some vaccines can elicit a concurrent reduction in parasite egg production. The expectation of a commercially feasible vaccine that might also reduce parasite transmission in the field is now realistic, although major hurdles still exist. Here, Terry Spithill and John Dalton review the results of several recent vaccine trials and discuss the future prospects for vaccine development.

  20. Age as a factor in acquisition of parasites by Canada geese

    USGS Publications Warehouse

    Wehr, E.E.; Herman, C.M.

    1954-01-01

    Examination of 46 Canada goose goslings yielded 14 species of parasites, including five Protozoa, four Nematoda, two Cestoda, and three Trematoda. Evidence indicates that goslings acquired most of these infections during their first week of life. Some parasites, Prosthogonimus sp., occurred only in younger birds. Others, Leucocytozoon simondi, were evident only during the initial course of infection, while still others remained evident in older geese. Parasites with a direct life cycle appeared to be more prevalent than those requiring intermediate hosts. Among 29 birds from a refuge in Michigan, 14 species of parasites were found; while in 17 goslings from a Utah refuge, only five species occurred.

  1. [Helminthofauna of wild canids in Azerbaijan and ways of its formation].

    PubMed

    Fataliev, G G

    2011-01-01

    The complete list of helminthes parasitizing canids in Azerbaijan is given; ways of formation of the canids' helminthofauna in Azerbaijan are reconstructed. As a result of our study, 42 helminth species were recorded; 25 of them parasitize jackals, 16 parasitize wolfs, and 39 species were found in foxes. The helminthofauna includes 5 species of Trematoda, 14 species of Cestoda, 1 species of Acanthocephala, and 22 species of Nematoda. By the life cycle, 32 species belong to biohelminthes and 10 species are geohelminthes. PMID:21874846

  2. Computer-aided procedure for counting waterfowl on aerial photographs

    USGS Publications Warehouse

    Bajzak, D.; Piatt, J.F.

    1990-01-01

    Examination of 46 Canada goose goslings yielded 14 species of parasites, including five Protozoa, four Nematoda, two Cestoda, and three Trematoda. Evidence indicates that goslings acquired most of these infections during their first week of life. Some parasites, Prosthogonimus sp., occurred only in younger birds. Others, Leucocytozoon simondi, were evident only during the initial course of infection, while still others remained evident in older geese. Parasites with a direct life cycle appeared to be more prevalent than those requiring intermediate hosts. Among 29 birds from a refuge in Michigan, 14 species of parasites were found; while in 17 goslings from a Utah refuge, only five species occurred.

  3. A new species of monorchiid (Digenea) from the South China Sea.

    PubMed

    Wang, Jiaxu; Wu, Wenzhong; Gao, Peng; Liu, Shengfa

    2013-06-01

    A new species of monorchiid trematode, Leiomonorchis zhanjiangensis n. sp. (Trematoda: Monorchiidae), is described from the intestine of marine fish Secutor ruconius (Hamilton-Buchanan) (Leiognathidae) from Zhanjiang (21°2'N, 110°3'E), Guangdong Province, China. The new species is distinguished from all known species of the genus, mainly by the configuration of vitelline follicles