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      Redescription of Clinostomum phalacrocoracis metacercariae (Digenea: Clinostomidae) in cichlids from Lake Kinneret, Israel Translated title: Redescription des métacercaires de Clinostomum phalacrocoracis (Digenea: Clinostomidae) de Cichlidae du Lac de Tibériade, Israël

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          Clinostomidae are digeneans characterized by a complex taxonomic history, continuously under revision based on both morphological and molecular analysis. Among the 14 species considered valid so far Clinostomum phalacrocoracis has been well described only at the adult stage, whereas the morphology of the metacercarial stage has been reported only once. During a parasitological survey carried out on 262 wild cichlids sampled from Lake Kinneret (Israel) metacercariae referable to C. phalacrocoracis were found in 18 fingerlings. In this study, we report this clinostomid species for the first time in wild fish from Israel describing the metacercarial stage of Clinostomum phalacrocoracis, coupling its morphological description with molecular analysis carried out on ITS rDNA and COI mtDNA sequences.

          Translated abstract

          Les Clinostomidae sont des digènes caractérisés par une histoire taxonomique complexe, constamment en cours de révision sur la base d’analyses morphologiques et moléculaires. Parmi les 14 espèces considérées comme valides, Clinostomum phalacrocoracis n’a été bien décrit qu’au stade adulte alors que la morphologie de la métacercaire n’a été rapportée qu’une seule fois. Pendant une enquête parasitologique réalisée sur 262 cichlidés sauvages prélevés dans le Lac de Tibériade (Israël), des métacercaires rapportées à C. phalacrocoracis ont été trouvées dans 18 poissons. Dans cette étude, nous présentons pour la première fois cette espèce de Clinostomidae chez les poissons sauvages en provenance d’Israël et redécrivons la métacercaire de Clinostomum phalacrocoracis, en associant sa description morphologique à une analyse moléculaire réalisée sur des séquences ITS (ADNr) et COI (ADNmt).

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          Most cited references 9

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          Development of primers for the mitochondrial cytochrome c oxidase I gene in digenetic trematodes (Platyhelminthes) illustrates the challenge of barcoding parasitic helminths.

          The phylum Platyhelminthes is a diverse group of flatworms that includes parasites with serious impacts on human health, animal husbandry, aquaculture and wildlife management. Here we present degenerate primers for the barcode region of the mitochondrial cytochrome c oxidase I (COI) gene in flatworms. Although amplicons were obtained from a wide taxonomic range in the Cestoda and Trematoda, COI fragments from many taxa in these classes did not amplify. Primers specific to trematodes in the family Diplostomidae were also developed. Amplification success was much higher with diplostomid-specific primers and sequences were obtained from 504 of 585 specimens of Diplostomum and Tylodelphys. Sequences from the barcode region resolved all specimens to the species level, with mean divergence between congeners of 19% (3.9-25%). Because many of our specimens were small, we initially amplified part of the nuclear small subunit ribosomal (r) RNA gene to evaluate the quality and quantity of DNA in our specimens. Short sequences (~380 nt) of this gene were recovered from most specimens and can be used to distinguish specimens at the family level and often the generic level. We suggest that rRNA genes could be used to screen samples of completely unknown taxonomy, after which specific COI primers could be used to obtain species-level identifications. © 2009 Blackwell Publishing Ltd.
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            Morphological and molecular differentiation of Clinostomum complanatum and Clinostomum marginatum (Digenea: Clinostomidae) metacercariae and adults.

            The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum , by ∼ 1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.
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              First description of the adult stage of Clinostomum cutaneum Paperna, 1964 (Digenea: Clinostomidae) from grey herons Ardea cinerea L. and a redescription of the metacercaria from the Nile tilapia Oreochromis niloticus niloticus (L.) in Kenya.

              The combined use of morphological and molecular studies allowed for the first time the recognition and description of the adult stage of Clinostomum cutaneum Paperna, 1964 from the grey heron Ardea cinerea L. in Kenya. A redescription of the metacercaria that infect Nile tilapia Oreochromis niloticus niloticus (L.) from the same aquatic environment is also presented. C. cutaneum differs from all other species of Clinostomum Leidy, 1856 in the shape of its uterus. Sequencing the rRNA confirmed the morphological similarity between adults from the grey heron and the metacercarial stage from tilapia, and a level of genetic similarity with the other previously sequenced Clinostomum spp. was observed. The need for a reorganisation of Clinostomum using both morphological and molecular methods is highlighted.

                Author and article information

                EDP Sciences
                03 July 2014
                : 21
                : ( publisher-idID: parasite/2014/01 )
                [1 ] Department of Veterinary Medical Sciences, Alma Mater Studiorum University of Bologna Via Tolara di Sopra 50 40064 Ozzano Emilia (BO) Italy
                [2 ] Central Fish Health Laboratory, Department of Fisheries and Aquaculture, Ministry of Agriculture and Rural Development 10803 Nir David Israel
                [3 ] Israel Oceanographic and Limnological Research, Kinneret Limnological Laboratory 14950 Migdal Israel
                Author notes
                [* ]Corresponding author: monica.caffara@ 123456unibo.it
                parasite140042 10.1051/parasite/2014034
                © M. Caffara et al., published by EDP Sciences, 2014

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 4, Tables: 0, Equations: 0, References: 29, Pages: 6
                Research Article


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