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

Particle and Fibre Toxicology

Aneta Kostadinova⁴  Héctor J Pula³  Simona Georgieva²  Ana Pérez-del-Olmo¹ 

[1] Unidad de Zoología Marina, Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Parc Científic, Universitat de València, Valencia 46071, Spain;Faculty of Science, University of South Bohemia, Brani¿ovská 31, ?eské Bud?jovice, 370 05, Czech Republic;Piscifactoría Vegas del Guadiana, Gobierno de Extremadura, Antigua Ctra. N-V, km. 391, 7 s/n, C.P. 06195, Villafranco del Guadiana, Badajoz, Spain;Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Brani¿ovská 31, ?eské Bud?jovice, 370 05, Czech Republic

关键词: ITS1-5.8S-ITS2;    Cox1;    Spain;    Gulls;    Freshwater fish;    Lens metacercariae;    Diplostomum pseudospathaceum;    Diplostomum spathaceum;   

Others  :  1149473
DOI  :  10.1186/s13071-014-0502-x

 received in 2014-09-27, accepted in 2014-10-25,  发布年份 2014

【 摘 要 】

Background

Recent molecular studies have revealed high species diversity of Diplostomum in central and northern Europe. However, our knowledge of the distribution of Diplostomum spp. in the southern distributional range in Europe of the snail intermediate hosts (Lymnaea stagnalis and Radix spp.) is rather limited. This study aims to fill this gap in our knowledge using molecular and morphological evidence.

Methods

Nineteen fish species and six fish-eating bird species were sampled opportunistically in three regions (Catalonia, Extremadura and Aragon) in Spain. All isolates of Diplostomum spp. were characterised morphologically and molecularly. Partial sequences of the barcode region of the cox1 mitochondrial gene and complete sequences of the ribosomal ITS1-5.8S-ITS2 gene cluster were used for molecular identification of the isolates.

Results

Integrated morphological and molecular analyses demonstrated the presence of three species among the larval and adult isolates of Diplostomum spp. sampled in Spain: Diplostomum spathaceum (in fish and birds), D. pseudospathaceum (in birds) and Diplostomum sp. (in fish) referred to as Clade Q sensu Georgieva et al. (Int J Parasitol, 43:57¿72, 2013). We detected ten cox1 haplotypes among the isolates of D. spathaceum with only one haplotype shared with adult isolates from central and northern Europe. No specific geographic pattern of the distribution of the novel haplotypes was found.

Conclusion

This first molecular exploration of the diversity of Diplostomum spp. in southern Europe indicates much lower species richness compared with the northern regions of Europe.

【 授权许可】

   
2014 Pérez-del-Olmo et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Georgieva S, Soldánová M, Pérez-del-Olmo A, Dangel RD, Sitko J, Sures B, Kostadinova A: Molecular prospecting for European Diplostomum (Digenea: Diplostomidae) reveals cryptic diversity. Int J Parasitol 2013, 43:57-72.
• [2]Jousson O, Bartoli P, Pawlowski J: Cryptic speciation among intestinal parasites (Trematoda: Digenea) infecting sympatric host fishes (Sparidae). J Evol Biol 2000, 13:778-785.
• [3]Georgieva S, Kostadinova A, Skirnisson K: The life-cycle of Petasiger islandicus Kostadinova & Skirnisson, 2007 (Digenea: Echinostomatidae) elucidated with the aid of molecular data. Syst Parasitol 2012, 82:177-183.
• [4]Born-Torrijos A, Kostadinova A, Raga JA, Holzer AS: Molecular and morphological identification of larval opecoelids (Digenea: Opecoelidae) parasitising prosobranch snails in a Western Mediterranean lagoon. Parasitol Int 2012, 61:450-460.
• [5]Alcántar-Escalera FJ, García-Varela M, Vázquez-Domínguez E, Pérez-Ponce de León G: Using DNA barcoding to link cystacanths and adults of the acanthocephalan Polymorphus brevis in central Mexico. Mol Ecol Resour 2013, 13:1116-1124.
• [6]Locke SA, McLaughlin JD, Dayanandan S, Marcogliese DJ: Diversity, specificity and evidence of hybridization in Diplostomum spp. metacercariae in freshwater fishes is revealed by DNA barcodes and ITS sequences. Int J Parasitol 2010, 40:333-343.
• [7]Blasco-Costa I, Faltýnková A, Georgieva S, Skírnisson K, Scholz T, Kostadinova A: Fish pathogens near the Arctic Circle: molecular, morphological and ecological evidence for unexpected diversity of Diplostomum (Digenea: Diplostomidae) in Iceland. Int J Parasitol 2014, 44:703-715.
• [8]Sanmartín ML, Cordeiro JA, Álvarez MF, Leiro JM: Helminth fauna of the yellow-legged gull Larus cachinnans in Galicia, north-west Spain. J Helm 2005, 79:361-371.
• [9]Álvarez MF, Cordeiro JA, Leiro JM, Sanmartín ML: Influence of host age and sex on the helminth fauna of the yellowlegged gull (Larus michahellis) in Galicia (Northwestern Spain). J Parasitol 2006, 92:454-458.
• [10]Ribas J, Miquel J, Torres J: New record of Diplostomum pseudospathaceum (Szidat, 1924) Niewiadomska, 1984 in Spain. Res Rev Parasitol 1999, 59:19-21.
• [11]Bosch M, Torres J, Figuerola J: A helminth community in breeding yellow-legged gulls (Larus cachinnans): pattern of association and its effect on host fitness. Can J Zool 2000, 78:777-786.
• [12]Aguilar A, Alvarez MF, Leiro JM, Sanmartín ML: Parasite populations of the European eel (Anguilla anguilla L.) in the Rivers Ulla and Tea (Galicia, northwest Spain). Aquaculture 2005, 249:85-94.
• [13]Behrmann-Godel J: Parasite identification, succession and infection pathways in perch fry (Perca fluviatilis): new insights through a combined morphological and genetic approach. Parasitology 2013, 140:509-520.
• [14]Pleijel F, Jondelius U, Norlinder E, Nygren A, Oxelman B, Schander C, Sundberg P, Thollesson M: Phylogenies without roots? A plea for the use of vouchers in molecular phylogenetic studies. Mol Phylogen Evol 2008, 48:369-371.
• [15]Georgieva S, Selbach C, Faltýnková A, Soldánová M, Sures B, Skírnisson K, Kostadinova A: New cryptic species of the `revolutum¿ group of Echinostoma (Digenea: Echinostomatidae) revealed by molecular and morphological data. Parasites & Vectors 2013, 6:64. BioMed Central Full Text
• [16]Moszczynska A, Locke SA, McLaughlin JD, Marcogliese DJ, Crease TJ: Development of primers for the mitochondrial cytochrome c oxidase I gene in digenetic trematodes illustrates the challenge of barcoding parasitic helminths. Mol Ecol Resour 2009, 9:75-82.
• [17]Galazzo DE, Dayanandan S, Marcogliese DJ, McLaughlin JD: Molecular systematics of some North American species of Diplostomum (Digenea) based on rDNA-sequence data and comparisons with European congeners. Can J Zool 2002, 80:2207-2217.
• [18]Luton K, Walker D, Blair D: Comparison of ribosomal internal transcribed spacers from two congeneric species of flukes (Platyhelminthes: Trematoda: Digenea). Mol Biochem Parasitol 1992, 56:323-328.
• [19]Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011, 28:2731-2739.
• [20]Telford MJ, Herniou EA, Russell RB, Littlewood DTJ: Changes in mitochondrial genetic codes as phylogenetic characters: Two examples from the flatworms. Proc Natl Acad Sci U S A 2000, 97:11359-11364.
• [21]Guindon S, Gascuel O: A simple, fast and accurate method to estimate large phylogenies by maximum-likelihood. Syst Biol 2003, 52:696-704.
• [22]Darriba D, Taboada GL, Doallo R, Posada D: jModelTest 2: more models, new heuristics and parallel computing. Nat Methods 2012, 9:772.
• [23]Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O: New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 2010, 59:307-321.
• [24]Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP: MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 2012, 61:539-542.
• [25]Huelsenbeck JP, Ronquist F, Nielsen R, Bollback JP: Bayesian Inference of phylogeny and its impact on evolutionary biology. Science 2001, 294:2310-2314.
• [26][http://www.iucnredlist.org/details/4785/0] webcite Anonymous: The IUCN Red List of threatened species. . Accessed on 10 July 2014.
• [27]Niewiadomska K: Present status of Diplostomum spathaceum (Rudolphi, 1819) and differentiation of Diplostomum pseudospathaceum nom. nov. (Trematoda: Diplostomatidae). Syst Parasitol 1984, 6:81-86.
• [28]Niewiadomska K: Verification of the life-cycles of Diplostomum spathaceum (Rudolphi, 1819) and D. pseudospathaceum Niewiadomska, 1984 (Trematoda: Diplostomidae). Syst Parasitol 1986, 8:23-31.
• [29]Niewiadomska K, Laskowski Z: Systematic relationships among six species of Diplostomum Nordmann, 1832 (Digenea) based on morphological and molecular data. Acta Parasitol 2002, 47:20-28.
• [30]Rellstab C, Louhi K-R, Karvonen A, Jokela J: Analysis of trematode parasite communities in fish eye lenses by pyrosequencing of naturally pooled DNA. Infect Genet Evol 2011, 11:1276-1286.