Combination of nuclear and mitochondrial markers as a useful tool to identify Ctenophthalmus species and subspecies (Siphonaptera: Ctenophthalmidae)

Antonio Zurita,Cristina Cutillas

doi:10.1007/s13127-021-00500-4

Abstract

Ctenophthalmus is considered the largest genus within the Order Siphonaptera. From a morphological point of view, only males of this genus can be identified at species and subspecies levels using morphological keys, whereas there are no morphological criteria in order to classify females at these taxonomical levels. Furthermore, the amount of available molecular and phylogenetic data for this genus is quite scarce so far. The main objective of this work was to assess the utility of the combination of nuclear and mitochondrial markers with respect to their ability to differentiate among different subspecies within the Ctenophthalmus genus. With this purpose, we carried out a comparative morphological and molecular study of three different subspecies (Ctenophthalmus baeticus arvernus, Ctenophthalmus nobilis dobyi, and Ctenophthalmus andorrensis catalaniensis) in order to clarify and discuss its taxonomic status. In addition, our study complemented the molecular data previously provided for Ctenophthalmus baeticus boisseauorum and Ctenophthalmus apertus allani subspecies. We sequenced five different molecular markers: EF1-α, ITS1, ITS2, cox1, and cytb. Our results confirmed that morphological data by themselves are not able to discriminate among Ctenophthalmus female taxa; however, the combination of the nuclear marker EF1-α together with mtDNA markers cytb and cox1 constituted a useful taxonomical and phylogenetic tool to solve this issue. Based on these results, we consider that the use of this molecular approach should be gradually used within Ctenophthalmus genus in order to complement its classical taxonomy and clarifying the complex taxonomy of other congeneric species of fleas.

Highlights

The family Ctenophthalmidae has been considered a “catchall” for a wide range of divergent taxa within the Order Siphonaptera (Whiting et al 2008; Keskin 2019)
Ctenophthalmus is considered the largest genus within the Order Siphonaptera; this genus comprises fifteen different subgenera distributed throughout the world out of which the Ctenophthalmus subgenus is the
The main objective of this work was to assess the utility of the combination of nuclear and mitochondrial markers with respect to their ability to differentiate among different subspecies within the Ctenophthalmus genus

Summary

Introduction

The family Ctenophthalmidae has been considered a “catchall” for a wide range of divergent taxa within the Order Siphonaptera (Whiting et al 2008; Keskin 2019). In this sense, some authors recently reported that some subfamilies included within Ctenophthalmidae such as Stenoponiinae should be considered a monophyletic family (Stenoponiidae) (Zurita et al 2015). Males of Ctenophthalmus can be distinguishable based on their complex genitalia In this context, it is usual to find some taxonomical keys for this genus providing a deep level of classification to subspecies level exclusively based on males (Beaucournu and Launay 1990). Beaucournu and Loverlec (2014) reported eight different

Objectives

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Conclusion