Whole genome amplification and sequencing of individual Dirofilaria immitis microfilariae.

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  • Additional Information
    • Source:
      Publisher: Academic Press Country of Publication: United States NLM ID: 0370713 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2449 (Electronic) Linking ISSN: 00144894 NLM ISO Abbreviation: Exp Parasitol Subsets: MEDLINE
    • Publication Information:
      Publication: Orlando, FL : Academic Press
      Original Publication: New York.
    • Subject Terms:
      Dirofilaria immitis*/genetics ; Dirofilaria immitis*/isolation & purification ; Genome, Helminth* ; Dog Diseases*/parasitology ; Dirofilariasis*/parasitology ; Microfilariae*/genetics ; Microfilariae*/isolation & purification ; Whole Genome Sequencing* ; DNA, Helminth*/isolation & purification ; DNA, Helminth*/chemistry; Animals ; Dogs ; Female ; Male
    • Abstract:
      Dirofilaria immitis is a filarial parasitic nematode of veterinary significance. With the emergence of drug-resistant isolates in the USA, it is imperative to determine the likelihood of resistance occurring in other regions of the world. One approach is to conduct population genetic studies across an extensive geographical range, and to sequence the genomes of individual worms to understand genome-wide genetic variation associated with resistance. The immature life stages of D. immitis found in the host blood are more accessible and less invasive to sample compared to extracting adult stages from the host heart. To assess the use of immature life stages for population genetic analyses, we have performed whole genome amplification and whole-genome sequencing on nine (n = 9) individual D. immitis microfilaria samples isolated from dog blood. On average, less than 1% of mapped reads aligned to each D. immitis genome (nuclear, mitochondrial, and Wolbachia endosymbiont). For the dog genome, an average of over 99% of mapped reads aligned to the nuclear genome and less than 1% aligned to the mitochondrial genome. The average coverage for all D. immitis genomes and the dog nuclear genome was less than 1, while the dog mitochondrial genome had an average coverage of 2.87. The overwhelming proportion of sequencing reads mapping to the dog host genome can be attributed to residual dog blood cells in the microfilariae samples. These results demonstrate the challenges of conducting genome-wide studies on individual immature parasite life stages, particularly in the presence of extraneous host DNA.
      Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
      (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: Canine heartworm; Dog; Filtration; Host contamination; Wolbachia
    • Accession Number:
      0 (DNA, Helminth)
    • Publication Date:
      Date Created: 20240715 Date Completed: 20240814 Latest Revision: 20240814
    • Publication Date:
      20250114
    • Accession Number:
      10.1016/j.exppara.2024.108806
    • Accession Number:
      39009178