Additional file 1 of Development and validation of a long-read metabarcoding platform for the detection of filarial worm pathogens of animals and humans

Supplementary Material

Additional file 1: of Awareness of zoonotic diseases and parasite control practices: a survey of dog and cat owners in Qatar

Figure S1. Multiple-choice questionnaire administered to dog and/or cat owners who attended the veterinary medical centres surveyed in the residential centre of Doha. (DOCX 121 kb

Assessment of the specificity of qPCR assay in the detection of <i>Onchocerca lupi</i> DNA.

<p>The amplification plot is represented by the fluorescent signal accordingly to relative fluorescence units (RFU) and threshold cycle.</p

Detection limit of the conventional PCR assay determined by 10-fold serial dilution of genomic DNA of microfilariae and adult of <i>Onchocerca lupi</i>.

<p>Lanes 1–4, from 3.6 ×10¹ pg/2μl to 3.6 ×10⁻³ pg/2μl of <i>O</i>. <i>lupi</i> mfs DNA (i.e., from 1 to 1×10⁻⁴ mfs); Lanes 5–15, from 8 ×10¹ ng/2μl to 8 x 10⁻³ fg/2μl of <i>O</i>. <i>lupi</i> adult DNA; Line 16, no-DNA control; M, 100 bp DNA marker.</p

A real-time PCR tool for the surveillance of zoonotic <i>Onchocerca lupi</i> in dogs, cats and potential vectors - Fig 2

<p>Standard curves generated from serial dilutions of (A) genomic DNA from adult (from 8 × 10⁴ to 8 × 10⁻³ fg/2μl of reaction) and microfilariae (B) (from 3.6 ×10⁻¹ ng/2μl to 3.6 ×10¹ fg/2μl of reaction) of <i>Onchocerca lupi</i>. Each point was tested in triplicate. Slope, efficacy and <i>R</i>² are reported on the bottom.</p

Skin samples tested for <i>Onchocerca lupi</i> by qPCR, divided (Groups 1–5) according to the parasitic load (mfs) microscopically detected.

<p>The mean, minimum, maximum and standard deviation (sd) values of the threshold cycle (Cq), parasite load (Starting Quantity (SQ) value, expressed as ng/μl of DNA for reaction) and microfilariae concentration, assessed by qPCR is reported.</p

GenBank accession numbers (AN) of mitochondrial cytochrome <i>c</i> oxidase subunit 1 sequences of filarial nematodes used for primers and TaqMan-probe design.

<p>GenBank accession numbers (AN) of mitochondrial cytochrome <i>c</i> oxidase subunit 1 sequences of filarial nematodes used for primers and TaqMan-probe design.</p

Filarial nematodes used to assess the analytical specificity of the qPCR assay.

<p>Filarial nematodes used to assess the analytical specificity of the qPCR assay.</p

Decreased ocular size lesions in dogs treated with oxfendazole.

<p>Decreased ocular size lesions in dogs n. 5 (A) and n. 9 (B) at 0 (D0), 90 (D90) and 180 (D180) days post enrolment.</p

Pharmacokinetic of oxfendazole in dogs treated for 10 days with oxfendazole.

<p>Mean (± s.e.m.) blood concentrations (μg/mL) of oxfendazole (square) and its metabolites fenbendazole (triangle) and fenbendazole-sulfone (triangle) following oral administration of 50 mg/kg b.w. for 10 days in dogs (n = 3).</p