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. 2025 Mar 26;12(1):280-286.
doi: 10.5455/javar.2025.l894. eCollection 2025 Mar.

Morphological and molecular identification of the cat flea Ctenocephalides felis from Bangladesh

Affiliations

Morphological and molecular identification of the cat flea Ctenocephalides felis from Bangladesh

Md Shamsudduha et al. J Adv Vet Anim Res. .

Abstract

Objective: The present study was designed to conduct molecular and morphological identification of cat fleas (Ctenocephalides felis) from Bangladesh along with nucleotide polymorphism and phylogenetic analysis.

Materials and methods: Samples were collected from two hosts (cat and human). The species was identified through morphological studies first, and then DNA was extracted for subsequent molecular analysis. A part of the mitochondrial 16S rRNA gene was amplified by polymerase chain reaction using extracted DNA as a template. The amplified region was sequenced using the Sanger dideoxy method. The sequence was subjected to NCBI BLASTn search. BioEdit and MEGA 11 software were used for multiple sequence alignment (MSA) and generating a phylogenetic tree.

Results: Morphological features such as shape, size, and appendages showed similarity with C. felis. The metatibial formula of chaetotaxy (2-2-2-2-1-3) was confirmed for species-level identification. NCBI BLASTn search showed the highest sequence identity with the available sequence of C. felis such as 99.78% (NC_049858.1) and 99.12% (MW420044.1 and MK941844.1) with 100% query coverage. MSA of C. felis sequences from different geographical distributions show their sequence affinities with each other, and the phylogenetic tree presents their relationship with each other.

Conclusion: Both morphological and molecular studies clearly indicate the identity and confirmation of cat flea (C. felis) from Bangladesh.

Keywords: 16S rRNA; Cat flea; Ctenocephalides felis; ectoparasite; molecular identification; morphological identification.

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Conflict of interest statement

The authors declared no potential conflicts of interest.

Figures

Figure 1.
Figure 1.. Female C. felis. (a) The left side of the whole body. (b) Head and thorax region, showing pointed frons; 1st genal ctenidial length exceeds half of the 2nd one; pronotal combs; and maxillary palpus. (c) Leg’s tibia bearing 5 notches; female spermatheca (a holding organ); tergal spiracles.
Figure 2.
Figure 2.. A partial view of the MSA of 16S rRNA region of C. felis from different countries. The scale on top represents site numbers.
Figure 3.
Figure 3.. Phylogenetic relationship among different species from the order Siphonaptera using mitochondrial 16S rRNA gene (a) and mitochondrial CO1 gene (b). *Marked sequence depicts the present study. Numbers representing bootstrap values.

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