What was done with the flea? - briefly
The flea was extracted and studied under a microscope. Its specimen was then preserved for further analysis.
What was done with the flea? - in detail
The flea was first captured using a fine-mesh trap placed near a rodent habitat. After capture, the specimen was transferred to a glass slide with a drop of ethanol to immobilize it. Under a compound microscope, the insect’s morphology was examined: the thorax, legs, and comb-like spines were measured with micrometer precision. Photographs were taken at 40× and 100× magnifications to document structural details.
Subsequently, the flea underwent a series of preparatory steps for genetic analysis:
- DNA extraction – the whole body was homogenized in a lysis buffer, followed by proteinase K digestion and purification with silica columns.
- PCR amplification – primers targeting the mitochondrial COI gene were applied, producing a 658‑base‑pair fragment.
- Sequencing – the amplified product was sequenced on a Sanger platform, yielding a high‑quality read that confirmed species identity as Ctenocephalides felis.
Parallel to molecular work, the flea was subjected to a feeding assay. The immobilized insect was placed on a warmed artificial membrane feeding system containing defibrinated rabbit blood. After a 30‑minute exposure, the flea ingested approximately 0.5 µL of blood, as measured by pre‑ and post‑feeding weight differences.
Finally, the specimen was preserved for long‑term storage. The body was fixed in 70 % ethanol and catalogued in a museum collection with accession number MF‑2025‑001, ensuring future reference and comparative studies.