1. bookVolume 50 (2013): Issue 2 (June 2013)
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22 Apr 2006
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access type Open Access

A polyphasic approach with molecular phylogeny for the characterization of Anisakis pegreffii (Anisakidae: Nematoda) in fishes from Adriatic Sea

Published Online: 31 Aug 2013
Page range: 112 - 118
Journal Details
License
Format
Journal
First Published
22 Apr 2006
Publication timeframe
4 times per year
Languages
English

In this study we investigated the morphometric and molecular characterization of a liver encapsulated third-stage larval population of Anisakis spp. infecting Merluccius merluccius and Lophius piscatorius caught in the Adriatic Sea waters (southern Italy). A polyphasic approach based on PCR-RFLP profiles of the ITS region, mitochondrial COI (cytochrome c oxidase subunit 1), sequencing and molecular phylogeny of ITS and mitochondrial COI was used to identify Anisakis larvae collected from fish samples. PCR-RFLP analysis showed three banding pattern corresponding to the peculiar pattern of A. pegreffii. Sequence data from ribosomal ITS and mitochondrial COI were analysed by Neighbour Joining, Minimum Evolution and Maximum Parsimony methods to evaluate the phylogenetic relationships among A. simplex sensu lato. The phylogenetic trees obtained for both ITS and COI revealed the existence of three distinct clades for A. simplex sensu sricto, A. simplex C and A. pegreffii and the sequences obtained in this study clearly clustered together with A. pegreffii sequences present in the database.

Keywords

[1] Abe, N., Ohya, N., Yanagiguchi, R. (2005): Molecular characterization of Anisakis pegreffii larvae in Pacific cod in Japan. J. Helminthol., 79: 303–306 http://dx.doi.org/10.1079/JOH2005290Search in Google Scholar

[2] Abollo, E., Paggi, L., Pascual, S., D’Amelio, S. (2003): Occurrence of recombinant genotypes of Anisakis simplex s.s. and Anisakis pegreffii (Nematoda:Anisakidae) in an area of sympatry. Infect Genet. Evol., 3: 175–181. http://dx.doi.org/10.1016/S1567-1348(03)00073-XSearch in Google Scholar

[3] Cavallero, S., Nadler, S. A., Paggi, L., Barros, N. B., D’Amelio, S. (2011): Molecular characterization and phylogeny of anisakid nematodes from cetaceans from southeastern Atlantic coasts of USA, Gulf of Mexico, and Caribbean Sea. Parasitol Res., 108: 781–792. DOI 10.1007/s00436-010-2226-y http://dx.doi.org/10.1007/s00436-010-2226-ySearch in Google Scholar

[4] Cavallero, S., Ligas, A., Bruschi, F., D’Amelio, S. (2012): Molecular identification of Anisakis spp. from fishes collected in the Tyrrhenian Sea (NW Mediterranean). Vet. Parasitol., 187: 563–566. DOI:10.1016/j.vetpar.2012.01.033 http://dx.doi.org/10.1016/j.vetpar.2012.01.033Search in Google Scholar

[5] Ceballos-Mendiola, G., Valero, A., Polo-Vico, R., Tèjada, M., Abattouy, N., Karl, H., De Las Heras, C., Martin-Sàanchez, J. (2010): Genetic variability of Anisakis simplex s.s. parasitizing European hake (Merluccius merluccius) in the Little Sole Bank area in the Northeast Atlantic. Parasitol. Res., 107: 1399–1404. DOI 10.1007/s00436-010-2009-5 http://dx.doi.org/10.1007/s00436-010-2009-5Search in Google Scholar

[6] Eisenback, J. D. (1985): Techniques for preparing nematodes for scanning electron microscopy. pp. 79–105. In: Barker, K. R., Coster, C. C., Sasser, J. N. (Eds). An Advanced Treatise of Meloidogyne Vol. II. Methodology. North Carolina State University and US. Agency for International Development, Raleigh, North Carolina Search in Google Scholar

[7] Johansen, (1940): Plant Microtechnique. New York, USA: McGraw-Hill Book Co. 523pp. Search in Google Scholar

[8] Larizza, A., Vovlas, N. (1995): Morphological observations on third-stage larvae of Anisakis simplex A (Anisakidae: Nematoda) from Adriatic and Ionian Waters. J. Helminthol. Society of Washington, 62: 260–264 Search in Google Scholar

[9] Johansen, (1940): Plant Microtechnique. New York, USA: McGraw-Hill Book Co. 523pp. Search in Google Scholar

[10] Mattiucci, S., Nascetti, G., Cianchi, R., Paggi, L., Arduino, P., Margolis, L., Brattey, J., Webb, S., D’Amelio, S., Orecchia, P., Bullini, L. (1997): Genetic and ecological data on Anisakis simplex complex with evidence for a new species (Nematoda, Ascaridoidea, Anisakidae). J. Parasitol., 83: 401–416 http://dx.doi.org/10.2307/3284402Search in Google Scholar

[11] Mattiucci, S., Nascetti, G. (2008): Adavances and trends in the molecular systematics of anisakid nematodes, with implications for their evolutionary ecology and hostparasite co-evolutionary processes. Adv. Parasitol., 66: 47–148 http://dx.doi.org/10.1016/S0065-308X(08)00202-9Search in Google Scholar

[12] Seinhorst, J. W. (1959): A rapid method for the transfer of nematodes from fixative to anhydrous glycerine. Nematologica, 4: 67–69 http://dx.doi.org/10.1163/187529259X00381Search in Google Scholar

[13] Tamura, K., Dudley, J., Nei, M., Kumar, S. (2007): MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24: 1596–1599 http://dx.doi.org/10.1093/molbev/msm092Search in Google Scholar

[14] Thompson, J. D., Higgins, D. G., Gibson, T. J. (1994): Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22: 4673–4680 http://dx.doi.org/10.1093/nar/22.22.4673Search in Google Scholar

[15] Umehara, A., Kawakami, Y., Matsui, T., Araki, J., Uchida, A. (2006): Molecular identification of Anisakis simplex sensu stricto and Anisakis pegreffii (Nematoda: Anisakidae) from fish and cetacean in Japanese waters. Parasitol. Int., 55: 267–271 http://dx.doi.org/10.1016/j.parint.2006.07.001Search in Google Scholar

[16] Vardic Smrzlic, I., Valic, D., Kapetanovic, D., Kurtovic, B., Teskeredzic, E. (2012): Molecular characterisation of Anisakidae larvae from fish in Adriatic Sea. Parasitol. Res., 111: 2385–2391. DOI 10.1007/s00436-012-3094-4 http://dx.doi.org/10.1007/s00436-012-3094-4Search in Google Scholar

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