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Salivary glands of the cat flea, Ctenocephalides felis: Dissection and microscopy guide.

ABSTRACT: Fleas are morphologically unique ectoparasites that are hardly mistaken for any other insect. Most flea species that feed on humans and their companion animals, including the cat flea (Ctenocephalides felis), have medical and veterinary importance. Besides facilitating blood acquisition, salivary biomolecules can modulate pathogen transmission. Thus, dissection of salivary glands is essential for comprehensive studies on disease vectors like the cat flea. Herein, we present the pictorial dissection protocol assisting future research targeting individual flea organs, for revealing their roles in vector competence and physiology. We provide a comprehensive guide, allowing researchers, even with limited practical experience, to successfully perform microdissection for collecting cat flea salivary glands. Furthermore, the protocol does not require expensive, sophisticated equipment and can be accomplished with routinely available tools. We illustrated expected results with morphological changes of salivary glands upon blood feeding as well as fluorescently stained these organs.

SUBMITTER: Danchenko M

PROVIDER: S-EPMC11016963 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature

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Salivary glands of the cat flea, <i>Ctenocephalides felis</i>: Dissection and microscopy guide.

Danchenko Monika M Macaluso Kevin R KR

Current research in insect science 20240404

Fleas are morphologically unique ectoparasites that are hardly mistaken for any other insect. Most flea species that feed on humans and their companion animals, including the cat flea (<i>Ctenocephalides felis</i>), have medical and veterinary importance. Besides facilitating blood acquisition, salivary biomolecules can modulate pathogen transmission. Thus, dissection of salivary glands is essential for comprehensive studies on disease vectors like the cat flea. Herein, we present the pictorial ...[more]

PMID: 38623392

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Project description:The hematophagous behaviour emerged independently in several instances during arthropod evolution. Survey of salivary gland and saliva composition and its pharmacological activity led to the conclusion that blood-feeding arthropods evolved a distinct salivary mixture that can interfere with host defensive response, thus facilitating blood acquisition and pathogen transmission. The cat flea, Ctenocephalides felis, is the major vector of several pathogens, including Rickettsia typhi, Rickettsia felis and Bartonella spp. and therefore, represents an important insect species from the medical and veterinary perspectives. Previously, a Sanger-based sialome of adult C. felis female salivary glands was published and reported 1,840 expressing sequence tags (ESTs) which were assembled into 896 contigs. Here, we provide a deeper insight into C. felis salivary gland composition using an Illumina-based sequencing approach. In the current dataset, we report 8,892 coding sequences (CDS) classified into 27 functional classes, which were assembled from 42,754,615 reads. Moreover, we paired our RNAseq data with a mass spectrometry analysis using the translated transcripts as a reference, confirming the presence of several putative secreted protein families in the cat flea salivary gland homogenates. Both transcriptomic and proteomic approaches confirmed that FS-H-like proteins and acid phosphatases lacking their putative catalytic residues are the two most abundant salivary proteins families of C. felis and are potentially related to blood acquisition. We also report several novel sequences similar to apyrases, odorant binding proteins, antigen 5, cholinesterases, proteases, and proteases inhibitors, in addition to putative novel sequences that presented low or no sequence identity to previously deposited sequences. Together, the data represents an extended reference for the identification and characterization of the pharmacological activity present in C. felis salivary glands.

Project description:BackgroundSaliva of hematophagous arthropods contains a diverse mixture of compounds that counteracts host hemostasis. Immunomodulatory and antiinflammatory components are also found in these organisms' saliva. Blood feeding evolved at least ten times within arthropods, providing a scenario of convergent evolution for the solution of the salivary potion. Perhaps because of immune pressure from hosts, the salivary proteins of related organisms have considerable divergence, and new protein families are often found within different genera of the same family or even among subgenera. Fleas radiated with their vertebrate hosts, including within the mammal expansion initiated 65 million years ago. Currently, only one flea species-the rat flea Xenopsylla cheopis-has been investigated by means of salivary transcriptome analysis to reveal salivary constituents, or sialome. We present the analysis of the sialome of cat flea Ctenocephaides felis.Methodology and critical findingsA salivary gland cDNA library from adult fleas was randomly sequenced, assembled, and annotated. Sialomes of cat and rat fleas have in common the enzyme families of phosphatases (inactive), CD-39-type apyrase, adenosine deaminases, and esterases. Antigen-5 members are also common to both sialomes, as are defensins. FS-I/Cys7 and the 8-Cys families of peptides are also shared by both fleas and are unique to these organisms. The Gly-His-rich peptide similar to holotricin was found only in the cat flea, as were the abundantly expressed Cys-less peptide and a novel short peptide family.Conclusions/significanceFleas, in contrast to bloodsucking Nematocera (mosquitoes, sand flies, and black flies), appear to concentrate a good portion of their sialome in small polypeptides, none of which have a known function but could act as inhibitors of hemostasis or inflammation. They are also unique in expansion of a phosphatase family that appears to be deficient of enzyme activity and has an unknown function.

Project description:BackgroundThe cat flea, Ctenocephalides felis, is prevalent worldwide, will parasitize animal reservoirs of plague, and is associated with human habitations in known plague foci. Despite its pervasiveness, limited information is available about the cat flea's competence as a vector for Yersinia pestis. It is generally considered to be a poor vector, based on studies examining early-phase transmission during the first week after infection, but transmission potential by the biofilm-dependent proventricular-blocking mechanism has never been systematically evaluated. In this study, we assessed the vector competence of cat fleas by both mechanisms. Because the feeding behavior of cat fleas differs markedly from important rat flea vectors, we also examined the influence of feeding behavior on transmission dynamics.Methodology/principal findingsGroups of cat fleas were infected with Y. pestis and subsequently provided access to sterile blood meals twice-weekly, 5 times per week, or daily for 4 weeks and monitored for infection, the development of proventricular biofilm and blockage, mortality, and the ability to transmit. In cat fleas allowed prolonged, daily access to blood meals, mimicking their natural feeding behavior, Y. pestis did not efficiently colonize the digestive tract and could only be transmitted during the first week after infection. In contrast, cat fleas that were fed intermittently, mimicking the feeding behavior of the efficient vector Xenopsylla cheopis, could become blocked and regularly transmitted Y. pestis for 3-4 weeks by the biofilm-mediated mechanism, but early-phase transmission was not detected.ConclusionsThe normal feeding behavior of C. felis, more than an intrinsic resistance to infection or blockage by Y. pestis, limits its vector competence. Rapid turnover of midgut contents results in bacterial clearance and disruption of biofilm accumulation in the proventriculus. Anatomical features of the cat flea foregut may also restrict transmission by both early-phase and proventricular biofilm-dependent mechanisms.

Dataset Information

Salivary glands of the cat flea, Ctenocephalides felis: Dissection and microscopy guide.

Publications

Salivary glands of the cat flea, <i>Ctenocephalides felis</i>: Dissection and microscopy guide.

Similar Datasets

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