Abstract
The phylogenetic relationships among 31 different flea isolates representing seven different species were studied by nucleotide sequence comparison of the internal transcribed spacer 1 (ITS1), internal transcribed spacer 2 (ITS2) and/or mitochondrial 16S ribosomal RNA gene (mt16S-rDNA) to examine the patterns of variation. Results show that all regions are useful in discriminating among flea species. In Ctenocephalides felis and Tunga penetrans, some differences in these gene regions occurred among different isolates within the same species. In the latter case, the differences are in the mt16S-rDNA region, with one isolate showing 48% divergence in nucleotide sequence. The taxonomic implications of this result are unclear at present. The gene regions revealed differences between C. felis isolates only after DNA sequencing the PCR products. Further differentiation among C. felis isolates was obtained using four different random binding primers (decamers) and primers for mammalian aldolase to amplify narrow differences in the genome. Using these primers we were able to discriminate between different C. felis isolates and determine that some of the genetic variation coincided with minor differences in response to the control agent imidacloprid. However, overall findings do not support the existence of subspecies of C. felis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baldwin B, Sanderson MJ, Porter JM, Wojciechowski MF, Campbell CS, Donoghue MJ (1995) The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny. Ann Mo Bot Gard 82:247–277
Coleman AW, Mai JC (1997) Ribosomal DNA ITS-1 and ITS-2 sequence comparisons as a tool for predicting genetic relatedness. J Mol Evol 45:68–177
Essig A, Rinder H, Gothe R, Zahler M (1999) Genetic differentiation of mites of the genus Chorioptes (Acari: Psoroptidae). Exp Appl Acarol 23:309–318
Haeselbarth E (1966) A note on the subspecies of C. felis in Africa south of the Sahara. Zool Anz 176:357–365
Haymer DS, McInnis DO (1994) Resolution of populations of the Mediterranean fruit fly at the DNA level using random primers for the polymerase chain reaction. Genome 37:244–248
Heise M, Epe C, Schnieder T (1999) Differences in the second internal transcribed spacer (ITS-2) of eight species of gastrointestinal nematodes of ruminants. J Parasitol 85:431–435
Hopkins GHE, Rothschild M (1953) An illustrated catalogue of the Rothschild collection of fleas in the British Museum, Vol 1. University Press, Cambridge
Krämer F, Mencke N (2001) Flea biology and control. Springer, Berlin Heidelberg New York
Lane RP, Crosskey RW (eds) (1993) Medical insects and arachnids. Chapman and Hall, London
Marcilla A, Bargues MD, Abad-Franch F, Panzera F, Carcavallo RU, Noireau F, Galvao C, Jurberg J, Miles MA, Dujardin JP, Mas-Coma S (2002) Nuclear rDNA ITS-2 sequences reveal polyphyly of Panstrongylus species (Hemiptera: Reduviidae: Triatominae), vectors of Trypanosoma cruzi. Infect Genet Evol 1:225–235
McIntosh AH, Grasela JJ, Matteri RL (1996) Identification of insect cell lines by DNA amplification fingerprinting (DAF). Insect Mol Biol 5:187–195
Mehlhorn H (ed) (2001) Encyclopedic references of parasitology. Springer, Berlin Heidelberg New York
Motazedian H, Noyes H, Maingon R (1996) Leishmania and Sauroleishmania: the use of random amplified polymorphic DNA for the identification of parasites from vertebrates and invertebrates. Exp Parasitol 83:150–154
Oliveros R, Cutillas C, De Rojas M, Arias P (2000) Characterization of four species of Trichuris (Nematoda: Enoplida) by their second internal transcribed spacer ribosomal DNA sequence. Parasitol Res 86:1008–1013
Parker A, Kornfield I (1996) An improved amplification and sequencing strategy for phylogenetic studies using the mitochondrial large subunit rRNA gene. Genome 39:793–797
Rinder H, Rausch RL, Takahashi K, Kopp H, Thomschke A, Löscher T (1997) Limited range of genetic variation in Echinococcus multilocularis. J Parasitol 83:1045–1050
Rust MK, Waggoner M, Hinkle NC, Mencke N, Hansen O, Vaughn M, Dryden MW, Payne P, Blagburn B, Jacobs DE, Bach T, Bledsoe D, Hopkins T, Mehlhorn H (2002) Development of a larval bioassay for the susceptibility of cat fleas (Siphonaptera: Pulicidae) to imidacloprid. J Med Entomol 39:671–674
Shouche YS, Patole MS (2000) Sequence analysis of mitochondrial 16S ribosomal RNA gene fragment from seven mosquito species. J Biosci 25:361–366
Vobis M, D’Haese J, Mehlhorn H, Feldmeier H, Heukelbach, Mencke N (2004) Molecular biological investigation of Brazilian Tunga penetrans isolates from man, dogs and cats. Parasitol Res (in press)
Whitfield JB, Cameron SA (1998) Hierarchical analysis of variation in the mitochondrial 16S rRNA gene among Hymenoptera. Mol Biol Evol 15:1728–1743
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Vobis, M., D’Haese, J., Mehlhorn, H. et al. Molecular phylogeny of isolates of Ctenocephalides felis and related species based on analysis of ITS1, ITS2 and mitochondrial 16S rDNA sequences and random binding primers. Parasitol Res 94, 219–226 (2004). https://doi.org/10.1007/s00436-004-1201-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-004-1201-x