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Molecular screening of nematodes in lacertid lizards from the Iberian Peninsula and Balearic Islands using 18S rRNA sequences

Published online by Cambridge University Press:  02 April 2012

A. Perera*
Affiliation:
CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Campus Agrário de Vairão, 4485-661 Vairão, Portugal
J.P.M.C. Maia
Affiliation:
CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Campus Agrário de Vairão, 4485-661 Vairão, Portugal
F. Jorge
Affiliation:
CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Campus Agrário de Vairão, 4485-661 Vairão, Portugal
D.J. Harris
Affiliation:
CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Campus Agrário de Vairão, 4485-661 Vairão, Portugal
*

Abstract

The development of molecular methods is becoming a promising field in the analysis of parasite fauna in wildlife species. This is especially useful in the case of parasite species where developmental larval stages are difficult to assess using standard methods. In this study we screened for the presence of parasitic nematodes infecting lacertid lizards from the Iberian Peninsula and Balearic islands using nematode-specific 18S ribosomal RNA gene primers on host tissue samples. Sequencing of positive samples revealed the presence of different genera of nematodes. The detection of Strongyloides, a monoxenous genus reported for the first time in Podarcis lilfordi is probably the result of the amplification of larval stages present in the host circulatory system. Two spirurid nematodes, Synhimantus and a new unidentified clade, were also found, suggesting that reptiles might be paratenic hosts of several spirurid species. This study demonstrates the benefits of using specific molecular markers on tissue samples to identify infecting larval stages of nematodes, otherwise difficult to assess using traditional screening methods.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2012 

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References

Acosta, I., Hernández, S., Gutiérrez, P.N., Martínez-Cruz, M.S., Hernández, E., Buffoni, L. & Martínez-Moreno, F.J. (2010) Acuaroid nematodes in the common kestrel (Falco tinnunculus) in the south of Spain. The Veterinary Journal 183, 234237.CrossRefGoogle ScholarPubMed
Anderson, R.C. (1984) The origins of zooparasitic nematodes. Canadian Journal of Zoology 62, 317328.CrossRefGoogle Scholar
Anderson, R.C. (Ed.) (2000) Nematode parasites of vertebrates. Their development and transmission. 2nd edn. 650 pp. London, CAB International.CrossRefGoogle Scholar
Anderson, R.C., Chabanaud, A.G. & Willmott, S. (Eds) (2009) Keys to the nematode parasites of vertebrates: Archival volume. 480 pp. New York, CAB International.CrossRefGoogle Scholar
Birova, V. & Calvo, A. (1977) Posibles vías de diseminación de nematodos parásitos de las gallinas. Revista Cubana de Ciencia Avícola 4, 5152.Google Scholar
Chabaud, A.G. (2009) Spirurida. pp. 334360in Anderson, R.C.Chabaud, A.G. & Willmott, S. (Eds) Keys to the nematode parasites of vertebrates. Archival volume. New York, CAB International.CrossRefGoogle Scholar
Dorris, M., De Ley, P. & Blaxter, M.L. (1999) Molecular analysis of nematode diversity and the evolution of parasitism. Parasitology Today 15, 188193.CrossRefGoogle ScholarPubMed
Dorris, M., Vincy, M.E. & Blaxter, M.L. (2002) Molecular phylogenetic analysis of the genus Strongyloides and related nematodes. International Journal of Parasitology 32, 15071517.CrossRefGoogle ScholarPubMed
Dos Santos, K.R., Carlos, B.C., Paduan, K.S., Kadri, S.M., Barrella, T.H., Amarante, M.R.V., Ribolla, P.E.M. & Da Silva, R.J. (2010) Morphological and molecular characterization of Strongyloides ophidiae (Nematoda, Strongyloididae). Journal of Helminthology 84, 136142.CrossRefGoogle ScholarPubMed
Drummond, A.J., Ashton, B., Buxton, S., Cheung, M., Cooper, A., Duran, C., Field, M., Heled, J., Kearse, M., Markowitz, S., Moir, R., Stones-Havas, S., Sturrock, S., Thierer, T. & Wilson, A. (2010) Geneious v5.3. Available at websitehttp://www.geneious.com (accessed accessed 1 May 2011).Google Scholar
Floyd, R.M., Rogers, A.D., Lambshead, J.D. & Smith, C.R. (2005) Nematode-specific PCR primers for the 18S small subunit rRNA gene. Molecular Ecology Notes 5, 611612.CrossRefGoogle Scholar
Gasser, R.B. (2001) Identification of parasitic nematodes and study of genetic variability using PCR approaches. pp. 5382in Kennedy, M.W. & Harnett, W. (Eds) Parasitic nematodes: Molecular biology, biochemistry and immunology. London, CAB International.CrossRefGoogle Scholar
Gibbons, L.M. (2009) Keys to the nematode parasites of vertebrates: Supplementary volume. 416 pp. New York, CAB International.CrossRefGoogle Scholar
Gil-Delgado, J.A., Verdejo, J. & Barba, E. (1995) Nestling diet and fledgling production of Eurasian kestrels (Falco tinnunculus) in eastern Spain. Journal of Raptor Research 29, 240244.Google Scholar
Grove, D.I. (Ed.) (1989) Strongyloidiasis: A major roundworm infection of man. 336 pp. London, Taylor and Francis.Google Scholar
Harris, D.J., Carranza, S., Arnold, E.N., Pinho, C. & Ferrand, N. (2002) Complex biogeographical distribution of genetic variation within Podarcis wall lizards across the Strait of Gibraltar. Journal of Biogeography 29, 12571262.CrossRefGoogle Scholar
Harris, D.J., Maia, J. & Perera, A. (2011) Molecular characterization of Hepatozoon species in reptiles from the Seychelles. Journal of Parasitology 97, 106110.CrossRefGoogle ScholarPubMed
Hasegawa, H., Hayashida, S., Ikeda, Y. & Sato, H. (2009) Hyper-variable regions in 18S rDNA of Strongyloides spp. as markers for species-specific diagnosis. Parasitology Research 104, 869874.CrossRefGoogle ScholarPubMed
Holt, P.E., Cooper, J.E. & Needham, J.R. (1979) Strongyloides infection in snakes: three case reports. The Veterinary Record 104, 213214.CrossRefGoogle ScholarPubMed
Honisch, M. & Krone, O. (2008) Phylogenetic relationships of Spiruromorpha from birds of prey based on 18S rDNA. Journal of Helminthology 82, 129133.CrossRefGoogle Scholar
Huelsenbeck, J.P. & Ronquist, F. (2001) MR-BAYES: Bayesian inference of phylogeny. Bioinformatics 17, 754755.CrossRefGoogle Scholar
Jobb, G., Von Haeseler, A. & Strimmer, K. (2004) TREEFINDER: a powerful graphical analysis environment for molecular phylogenetics. BMC Evolutionary Biology 4, 18.CrossRefGoogle ScholarPubMed
Jones, R., Brown, D.S., Harris, E., Jones, J., Symondson, W.O.C., Bruford, M.W. & Cable, J. (2012) First record of Neoxysomatium brevicaudatum through the non-invasive sampling of Anguis fragilis: complementary morphological and molecular detection. Journal of Helminthology 86, 125129.CrossRefGoogle ScholarPubMed
Jorge, F., Roca, V., Perera, A., Harris, D.J. & Carretero, M.A. (2011) A phylogenetic assessment of the colonization patterns in Spauligodon atlanticus Astasio-Arbiza et al., 1987 (Nematoda: Oxyurida: Pharyngodonidae), a parasite of lizards of the genus Gallotia Boulenger: no simple answers. Systematic Parasitology 80, 5366.CrossRefGoogle ScholarPubMed
Maia, J.P.M.C., Harris, D.J. & Perera, A. (2011) Molecular survey of Hepatozoon species in lizards from North Africa. Journal of Parasitology 97, 513517.CrossRefGoogle ScholarPubMed
Pafilis, P., Meiri, S., Foufopoulos, J. & Valakos, E. (2009) Intraspecific competition and high food availability are associated with insular gigantism in a lizard. Naturwissenschaften 96, 11071113.CrossRefGoogle Scholar
Pereira, C. (1929) Strongyloides ophididae n. sp. Boletim de Biologia de Sao Paulo 15, 1617.Google Scholar
Perera, A. & Harris, D.J. (2010) Genetic variability within the Oudri's fan-footed gecko Ptyodactylus oudrii in North Africa assessed using mitochondrial and nuclear DNA sequences. Molecular Phylogenetics and Evolution 54, 634639.CrossRefGoogle ScholarPubMed
Pinho, C., Ferrand, N. & Harris, D.J. (2006) Reexamination of the Iberian and North African Podarcis (Squamata: Lacertidae) phylogeny based on increased mitochondrial DNA sequencing. Molecular Phylogenetics and Evolution 38, 266273.CrossRefGoogle ScholarPubMed
Roca, V. & Hornero, M.J. (1992) Strongyloides ophiusensis sp. n (Nematoda: Strongyloididae), parasite of an insular lizard, Podarcis pityusensis (Sauria, Lacertidae). Folia Parasitologica 39, 369373.Google Scholar
Rodrigues, O.H. (1968) Sobre nova especie do genero Strongyloides Grassi, 1879 (Nematoda, Rhabdiasoidea). Atas Sociedade de Biologia 12, 3132.Google Scholar
Salvador, A. (Ed.) (1997) Fauna Ibérica: Reptiles, vol. 10, 705 pp. Madrid, Museo Nacional de Ciencias Naturales, Centro Superior de Investigaciones Científicas.Google Scholar
Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular cloning: A laboratory manual. 545 pp. New York, Cold Spring Harbor Press.Google Scholar
Sanmartín, M.L., Álvarez, F., Barreiro, G. & Leiro, J. (2004) Helminth fauna of Falconiform and Strigiform birds of prey in Galicia, Northwest Spain. Parasitology Research 92, 255263.CrossRefGoogle ScholarPubMed
Speare, R. (1989) Identification of species of Strongyloides. pp. 1184in Grove, D.I. (Ed.) Strongyloidiasis: a major roundworm infection of man. London, Taylor and Francis.Google Scholar
Thompson, J.D., Higgins, D.G. & Gibson, T.J. (1994) Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acid Research 22, 46734680.CrossRefGoogle ScholarPubMed
Viney, M.E. & Lok, J.B. (2007) Strongyloides spp. In WormBook. The online review of C. elegans biology. Available at websitehttp://www.wormbook.org (accessed accessed 2 March 2012).CrossRefGoogle Scholar