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February 2, 2026Animals2 citationsOpen Access

Low Zoonotic Pathogen Burden in Free-Roaming Cats Revealed by 18S rRNA Metabarcoding: A Baseline Study from an Insular Natura 2000 Site in Spain

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MTMaría del Mar Travieso-AjaLHLuis Alberto Henríquez-HernándezEHElisa Hernández-Álvarez

Key Points

  • The aim is to establish a baseline profile of potentially pathogenic eukaryotes in free-roaming cats before antiparasitic interventions.
  • Conducted a cross-sectional 18S rRNA metabarcoding survey of community cats' faecal samples.
  • Collected 152 faecal samples, including fresh and dry deposits, during a TNR campaign.
  • Used a feline 18S blocking primer to enhance host amplification.
  • Sequenced libraries with Oxford Nanopore technology and assigned taxonomy using SILVA-based classifiers.
  • Detected DNA from at least 24 potentially pathogenic taxa among the studied samples.
  • Dipylidium caninum was the most frequent pathogen, found in 74.3% of samples.
  • Fresh samples had a higher richness of potentially pathogenic taxa compared to dry samples (p < 0.01).
  • Zoonotic protozoa were present but at low-to-moderate frequencies, with Toxoplasma gondii found in 7.9% of samples.

Abstract

Free-roaming cats may contribute to zoonotic risk via parasites and other eukaryotic taxa, yet surveillance in protected island settings is limited and conventional coprology can miss low-intensity or degraded signals. We conducted a cross-sectional 18S rRNA metabarcoding survey to establish a baseline profile of potentially pathogenic eukaryotes in community cats from La Graciosa (Natura 2000, Canary Islands, Spain) prior to large-scale antiparasitic interventions. We analysed 152 faecal samples, including fresh samples collected during a high-throughput TNR campaign (n = 37) and dry environmental deposits (n = 115). Host amplification was reduced using a feline 18S blocking primer; libraries were sequenced with Oxford Nanopore technology; and taxonomy was assigned using SILVA-based classifiers with downstream filtering for veterinary/zoonotic relevance. After quality control, 72 eukaryotic taxa were retained and DNA from at least 24 potentially pathogenic taxa was detected. Dipylidium caninum was most frequent (74.3%; 113/152), and opportunistic fungi/yeasts were common (e.g., Pichia kudriavzevii 42.4%, Diutina catenulata 31.5%). Zoonotic protozoa showed low-to-moderate detection frequency (Acanthamoeba castellanii 13.3%, Toxoplasma gondii 7.9%, Balamuthia mandrillaris 4.6%). Overall richness did not differ between fresh and dry samples (p > 0.05), but fresh samples contained higher richness of potentially pathogenic taxa (p < 0.01).

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Cite This Study

Travieso-Aja et al. (2026) studied this question.

synapsesocial.com/papers/6980fc55c1c9540dea80e253https://doi.org/10.3390/ani16030431
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