Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: Larval development assays were undertaken for all lines, at pre- and post-treatment timepoints, in the third generation of selection. (A) Control (CTL) lines had no treatment applied but sampling was time-matched with the selected lines ((B) IVM = ivermectin and (C) MOX = moxidectin). Y axis shows the proportion of the population in each well that developed to L3.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Selection, Control, Sampling

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A–C) Each sample from each generation is compared with the F0 parental sample and 10 kb window F st values plotted along the chromosomes (I-X). For each of the ivermectin selected (IVM) and moxidectin selected (MOX) lines, a line was plotted indicating five standard deviations above the genome-wide mean of the F3 generations (dashed and dotted respectively). CTL = control lines. Note that no sequence data was available for sample IVM1. (D) Chromosome V pairwise comparisons of third generation ivermectin and moxidectin samples to each other. Horizontal lines are five standard deviations above the mean F st value for the F3 generations – the same as in panels A-C, but with both ivermectin and moxidectin lines overlaid on each faceted plot.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Genome Wide, Control, Sequencing

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A–C) Ratio of nucleotide diversity in three generations (F1 to F3) of sub-therapeutic selection in comparison to F0 sample population. Key: C = control, I = ivermectin, M = moxidectin. L1 to L3 indicate biological replicate selection lines. Pi ratio = Normalised Sample θπ/ Normalised F0 θπ. Nucleotide diversity for each sample window was normalised by dividing by the genome-wide median diversity. Colour indicates the rank within the treatment group, where blue indicates a higher diversity than F0, and red a lower diversity. Note that the rank indicates the genome-wide ranking. Note that no sequence data was available for sample F1_I_L1. (D) Nucleotide diversity (θπ) of the F0 sample along Chromosome V. More negative values indicate lower diversity.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Selection, Comparison, Control, Genome Wide, Sequencing

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A) Ivermectin selected lines, (B) moxidectin selected lines, (C) control lines, (D) F0 sample. Note that no sequence data was available for sample F1_I_L1. More negative values (red) correlate to more rare alleles than expected for the population size, while more positive values (blue) indicate fewer rare alleles than expected and either very few alleles at all (highly conserved), or balancing selection of alleles. If the population is neutrally evolving, Tajima’s D is close to zero (mix of rare and common alleles).

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Control, Sequencing, Selection

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: Negative values indicate a reduction in the reference allele frequency at that site from F0 to F3. Each point is a SNP; red points are those which are significantly different to controls following Holm’s correction of the p-values, and where the corrected p-value is < 0.001. In other words: if red, the reference allele is changing in treated lines differently to control lines, and if negative it is reducing in the treated lines. (A) ivermectin vs control lines (slope coefficient plotted for ivermectin lines), (B) moxidectin vs control lines (slope coefficient plotted for moxidectin lines), (C) selected (ivermectin and moxidectin) vs control lines (slope coefficient plotted for combined selected lines).

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Control

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: PCA plots for male (A) and female (B) datasets. Genome-wide differential gene expression visualised as karyoplots (C) with i. ivermectin selected males versus control males, ii. ivermectin selected females versus control females, iii. moxidectin selected males versus control males, iv. moxidectin selected females versus control females. Upregulated genes are in yellow, downregulated genes are in blue, with adjusted P < 0.01 for significance.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Genome Wide, Gene Expression, Control

Journal: MethodsX

Article Title: A quick and simple method for the determination of ivermectin in dog plasma by LC–MS/MS

doi: 10.1016/j.mex.2018.11.011

Figure Lengend Snippet: Chromatograms of a blank sample (A), a blank sample spiked at the LLOQ (0.5 ng/mL) (B), and an actual dog sample collected 240 min after ivermectin oral administration (C), obtained by LC–MS/MS analysis. Early-eluting matrix contaminants and the last part of the LC run were diverted to waste through a Valco ® divert valve.

Article Snippet: Ivermectin chemical reference substance (CRS) was supplied by the European Directorate for the Quality of Medicines and HealthCare (EDQM).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: MethodsX

Article Title: A quick and simple method for the determination of ivermectin in dog plasma by LC–MS/MS

doi: 10.1016/j.mex.2018.11.011

Figure Lengend Snippet: Intraday and interday accuracy (D% = percentage difference between the experimental and theoretical values) and precision (CV%) for IVM (ivermectin), calculated on data obtained analyzing QC (quality control) samples prepared in three different days (total n = 54).

Article Snippet: Ivermectin chemical reference substance (CRS) was supplied by the European Directorate for the Quality of Medicines and HealthCare (EDQM).

Techniques: Control

Journal: MethodsX

Article Title: A quick and simple method for the determination of ivermectin in dog plasma by LC–MS/MS

doi: 10.1016/j.mex.2018.11.011

Figure Lengend Snippet:

Article Snippet: Ivermectin chemical reference substance (CRS) was supplied by the European Directorate for the Quality of Medicines and HealthCare (EDQM).

Techniques: Clinical Proteomics