Whole genome amplification and sequencing of individual Dirofilaria immitis microfilariae.

Exp Parasitol

Sydney School of Veterinary Science, Faculty of Science, The University of Sydney, New South Wales, 2006, Australia; Sydney Infectious Diseases Institute, The University of Sydney, New South Wales, 2006, Australia. Electronic address:

Published: August 2024

AI Article Synopsis

  • Dirofilaria immitis is a significant veterinary parasite, and the rise of drug-resistant strains in the USA highlights the need to investigate resistance in other regions through global population genetic studies and genome sequencing.
  • Researchers focused on immature life stages of D. immitis, which are easier to sample from dog blood than adult worms, conducting whole-genome amplification and sequencing on nine microfilaria samples.
  • The study found that less than 1% of sequenced reads matched D. immitis genomes due to a high prevalence of dog DNA in the samples, showcasing the difficulties of genome-wide studies on parasitic life stages amid host DNA contamination.

Article 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.

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http://dx.doi.org/10.1016/j.exppara.2024.108806DOI Listing

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