Molecular and morphological differentiation of Pulex irritans (Siphonaptera: Pulicidae) collected from Spain and Argentina. Distinct subspecies or genotypic plasticity?

ZURITA ANTONIO; MARCELA LARESCHI; URDAPILLETA MARA; CALLEJON M; CUTILLAS CRISTINA

Tenerife

Congreso; XX SOCEPA (Congreso de la Sociedad Española de Parasitología) & XV Meeting of the European Veterinary Parasitology College; 2017

XX SOCEPA (Congreso de la Sociedad Española de Parasitología) & XV Meeting of the European Veterinary Parasitology College

ObjetiveThe aim of this work was to carry out a comparative morphological and molecular study of P. irritans isolated from two different geographical regions: Seville (Spain) and Santa Cruz (Argentina) by sequencing different nuclear and mitochondrial molecular markers and considering morphological characteristics cited by previous authors.ResultsFifty-five fleas were collected from an abandoned horse stable from Seville (southwestern of Spain) and twenty-five fleas were isolated from Andean foxes (Lycalopex culpaeus) from Santa Cruz (South of Argentina).All specimens isolated in this work showed specific morphological characteristics of P. irritans. Furthermore, we did not find significant morphological differences between both geographical origins. The Internal Transcribed Spacers 1 and 2 (ITS1 and ITS2), partial 18S rRNA gene, partial cytochrome c-oxidase 1 (cox1) and cytochrome b (cytb) mtDNA gene sequences were determined to clarify the taxonomic status of these taxa and to assess intra-specific and intra-population similarity. According to ITS´s analyses, ITS2 sequences of both geographical origins were identical in length and markedly shorter than ITS1 sequences. However, ITS1 sequences showed differences in length according to geographical origins. Percentages of intra-specific similarity showed different values according to the molecular marked used. Thus, based on 18S and ITS2 sequences analyses we observed higher values of similarity between both geographical origins (100 % and 99.1 %, respectively), than those obtained when we analysed ITS1, cox1 and cytb sequences (94.6 %, 92 % and 91.8 %, respectively). On the other hand based on cox1 and cytb sequences, restriction mapping identified endonucleases delineating the two different geographical areas (Spain and Argentina). Furthermore, based on phylogenetic ITS1, ITS2, cox1, cytb and concatenated trees we observed individuals classified as P. irritans clustered clearly separated according to different geographical origins (Spain and Argentina).ConclusionsOn the basis on phylogenetic and molecular comparative study of two nuclear markers (ITS1 and ITS2), two mitochondrial markers (cox1 and cytb) and concatenated sequences we reported the existence of two geographical genetic lineages in P. irritans which could correspond with two different subspecies. Furthermore, both geneticlineages could be easily discriminated by PCR-linked random-fragment-length polymorphism (RFLP).