Morphological, biometrical, and molecular characterization of Ctenocephalides felis and Ctenocephalides canis isolated from dogs from different geographical regions

Abstract

In the present work, a comparative morphological, biometrical and molecular study of Ctenocephalides spp. isolated from dogs (Canis lupus familiaris) from different geographical regions (Spain, Iran, and South Africa) has been carried out. The internal transcribed spacer 1 (ITS1) sequences of Ctenocephalides felis and Ctenocephalides canis collected from dogs from different geographical regions have been determined to clarify the taxonomic status of these species and to assess intraspecific variation and interspecific sequence differences. In addition, a phylogenetic analysis based on ITS1 sequences has been performed. Four different morphological populations were observed in the individuals of C. felis collected from dogs from different geographical locations. Nevertheless, the comparative study of the ITS1 sequences of the different morphological populations observed in C. felis did not show molecular differences. The results showed clear molecular differences between C. felis and C. canis and some specific recognition sites for endonucleases were detected between both species. Thus, BfrBI and DraI sites have diagnostic value for specific determination in C. felis. The phylogenetic tree based on the ITS1 sequences of C. felis and C. canis revealed that all the populations of C. felis from different geographical regions clustered together and separated, with high bootstrap values, from C. canis. We conclude that ITS1 region is a useful tool to approach different taxonomic and phylogenetic questions in Ctenocephalides species.

Ctenocephalides felis and Ctenocephalides canis: introgressive hybridization?

“…() and Marrugal et al . (), who found that the ITS1 nucleotide sequences of different C. felis populations from different geographical areas are practically identical. Nevertheless, other authors such as Gamerschlag et al .…”

“…All specimens were photographed. Morphological differentiation between C. felis and C. canis individuals was carried out according to the original descriptions (Gil Collado, , ; Beaucournu & Launay, ; Lewis, ; Beaucournu & Ménier, ; Ménier & Beaucournu, ; Marrugal et al , ).…”

“…Several criteria cited by different authors were used for the specific determination of the genus Ctenocephalides including: Genal ctenidium formed of eight or nine spines oriented horizontally (Lewis, ) and relative size of the first and second genal spines (Gil Collado, ; Beaucournu & Ménier, ; Durden & Traub, ; Marrugal et al , ). Shape of the front of head (Gil Collado, ; Beaucournu & Launay, ; Lewis, ; Linardi & Santos, ; Marrugal et al , ). Length/width ratio of the head (Durden & Traub, ; Marrugal et al , ). Male genitalia: manubrium and apex (degree of dilation) (Gil Collado, ; Lewis, ; Ménier & Beaucournu, ; Marrugal et al , ) and aedeagus (Ménier & Beaucournu, ; Marrugal et al , ). Female genitalia: spermatheca and hilla (degree of elongation of the apical part of the spermatheca) (Gil Collado, ; Lewis, ; Marrugal et al , ). Presence of two to three bristles on the lateral metanotal area (LMA) (Gil Collado, ; Beaucournu & Launay, ; Beaucournu & Ménier, ; Linardi & Santos, ; Marrugal et al , ). Hind tibia with a number of seta‐bearing notches along dorsal margin (Beaucournu & Launay, ; Lewis, ; Beaucournu & Ménier, ; Durden & Traub, ; Linardi & Santos, ; Marrugal et al , ). …”

“…From a morphological point of view, four subspecies of C. felis have been distinguished: C. felis felis Bouché, 1835, C. felis strongylus Jordan, 1925, C. felis orientis Jordan, 1925 and C. felis damarensis Jordan, 1936 (Hopkins & Rothschild, ). However, only a few studies have been carried out based on molecular data (Vobis et al , ; Marrugal et al , ; Lawrence et al , , ). Thus, some authors have suggested that further molecular studies are needed in order to resolve and elucidate the genetic diversity of C. felis (Lawrence et al , ).…”

“…The ribosomal DNA segments Internal Transcribed Spacer 1 and 2 (ITS1 and 2) have been shown to be two of the best molecular markers for analysing genetic relationships at the species level in arthropods (De Rojas et al , , ; Marcilla et al , ; Monje et al , ). Recently, Marrugal et al () concluded that phylogenetic analysis based on the ITS1 region is a useful tool to approach different taxonomic and phylogenetic issues in Ctenocephalides species, and they found clear molecular differences between C. felis and C. canis . In addition, the 18S rRNA gene has been widely used as a molecular marker in order to clarify molecular relationships among families and subfamilies (Whiting et al , ; Acosta & Morrone, ; Díaz‐Nieto et al , ).…”