Anopheles mosquitoes exposed to long-acting antimalarials via drug-spiked supplemented bloodmeal absorb drug but do not suffer fitness costs
Data files
Jun 23, 2026 version files 115.28 KB
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Goheen_Impact_of_antimalarials_in_mosquitoes_Dryad.xlsx
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README.md
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Abstract
Background: The World Health Organization's recommendations for antimalarial chemoprevention and mass treatment use in endemic areas have greatly expanded, allowing more flexibility in demographic groups and regions targeted. An overlooked aspect of expanding human population-level drug exposure is the downstream impact of drug on the vector. Anopheles mosquitoes re-feed often with ≥4 bloodmeals during their lifespan. This provides repeated opportunities for mosquitoes to ingest drug via bloodmeals taken from people with antimalarials in the bloodstream and raises questions as to drug impact on mosquitoes.
Methods: We supplemented bloodmeals with physiologic levels of long-acting antimalarials to assess drug exposure impact on Anopheles mosquito fitness and determine drug absorption extent.
Results: We did not observe any significant differences in mosquito feeding, behavior, fecundity, or viability after amodiaquine, desethylamodiaquine, piperaquine, and sulfadoxine-pyrimethamine ingestion in lab-reared An. gambiae or field-derived An. coluzzii mosquitoes. Interrogating drug distribution within mosquitoes utilizing LC-MS/MS, desethylamodiaquine, amodiaquine’s longer-acting active metabolite, was fed at half and twice Cmax concentrations, with drug then dose-dependently detected in pooled whole mosquitoes, midguts, and hemolymph. This was significant for whole mosquitoes harvested at 24hrs and 120hrs, and midguts harvested at 24hrs. Between 24 to 120hrs, drug decreased in midguts but increased in hemolymph.
Conclusions: We show sustained biochemical evidence of antimalarial absorption into Anopheles hemolymph following bloodmeal ingestion without fitness costs. These studies provide a foundation to next assess vector-stage antimalarial drug exposure impact on parasite development in infected mosquitoes, which could in turn have implications on transmission dynamics and drug resistance spread.
Dataset DOI: 10.5061/dryad.pvmcvdp1h
Description of the data and file structure
Dryad data descriptions:
The data contains results from experiments whereby lab-derived An. gambiae and field-derived An. coluzzii mosquitoes were offered bloodmeals supplemented with solvents and tested antimalarials. Specifically, plasma was supplemented with DMSO, acetonitrile (ACN):H2O 1:1 v/v + 0.5% formic acid (FA), ACN:H2O 1:9 v/v + 0.5% FA, ACN:H2O 1:1, FA 0.5%, ACN:H2O 1:1 + 0.1% HCl, and 0.1% HCl, all used at 1:833 dilution in plasma and mixed with RBCs at 40% hematocrit in initial solvent control experiments using lab-derived An. gambiae. In further experiments with both lab An. gambiae and field An. coluzzii, plasma or serum, respectively, was supplemented with the antimalarials amodiaquine (AQ), desethylamodiaquine (DEAQ), piperaquine (PPQ), or sulfadoxine (SDX) and pyrimethamine (PYR) (SP combined) at levels ranging from one half (1/2X) to twice (2X) the average reported Cmax of standard human dosing, or with the endectocide IVM (at 1X Cmax), and combined with RBCs at 40% hematocrit for mosquito feeds. (Per available literature, Cmax was approximated as 20 ng/ml for AQ, 500 ng/ml for DEAQ, 300 ng/ml for PPQ, 100 ɥg/ml for SDX, 400 ng/ml for PYR, 50 ng/ml for IVM.) Negative controls included un-supplemented blood (“Blood only”) or for the drug feeding experiments, blood supplemented with the relevant drug solvent alone (“Solvent”) for each drug group. Solvents used in the drug feeding experiments were: ACN:H2O 1:9 + 0.5% FA for AQ, DEAQ, PPQ; ACN:H2O 1:1 + 0.1% HCl for SDX; ACN:H2O 1:1 + 0.5% FA for PYR (for SP experiments, “Solvent” represents the combination of both SDX and PYR solvents). Anopheles mosquitoes were 5-10 days old Anopheles upon feeding. Approximately 50 mosquitoes were fed per group for each experimental replicate.
Date of data collection: (2023-09-10 to 2024-12-27)
Geographic location of data collection: Connecticut, USA; Bobo-Dioulasso, Burkina Faso
Further descriptions for each data set are included below (with bolded words corresponding to Excel tab labels):
Feeding rates for lab-derived An. gambiae and field-derived An. coluzzii mosquitoes offered bloodmeals supplemented with solvents and tested antimalarials. Data represent the percent of female mosquitoes that fed (abdomen with visible bloodmeal) under each condition, reported as the mean percent fed between experimental replicates (and Standard Deviation) (two replicates for initial solvent control feeding; three replicates for all drug feeding experiments).
Raw feeding data and contingency analyses for lab-derived An. gambiae and field-derived An. coluzzii mosquitoes offered bloodmeals supplemented with solvents and tested antimalarials. Data represent the number of female mosquitoes that fed (abdomen with visible bloodmeal) or did not feed under each condition, reported as the total between experimental replicates (two replicates for initial solvent control feeding; three replicates for all drug feeding experiments other than four replicates for the experiment with AQ/DEAQ field mosquitoes). Contingency analyses comparing the rates of fed to unfed mosquitoes for each experimental condition are included against the blood only or relevant solvent controls (reported as p values, italicized if <0.05 with direction of significance stated to the right of any significant p values). Contingency analysis is also included at the bottom for total lab An. gambiae and field An. coluzzii feeding rates under all conditions.
Percent survival of lab-derived An. gambiae and field-derived An. coluzzii mosquitoes following ingestion of bloodmeal supplemented with solvents or tested antimalarials. Data represent percent survival of mosquitoes (unfed mosquitoes were removed; exact n is the number that fed for each experimental replicate minus 5 that were separated out to monitor fertility) over time (days) after feeding on bloodmeals supplemented with solvents and drugs of interest. Data are from 3 experimental replicates unless otherwise noted, with individual data for each time/condition shown. (The conditions which other than 3 replicates exist include: individual null labels for few times/conditions when data was not captured; 2 replicates for initial solvent control experiments; 4 replicates for An. coluzzii AQ/DEAQ feeds; 9 replicates combining all An. gambiae IVM feeding data and 10 replicates combining all An. coluzzii IVM feeding data.) Mean percent alive (and Standard Deviation) are also shown for each time/condition.
Survival curve comparisons for lab-derived An. gambiae and field-derived An. coluzzii mosquitoes following ingestion of bloodmeal supplemented with solvents and tested antimalarials. Data represent Kaplan-Meier survival curve comparisons between specified groups, with Chi square results and p values (unadjusted and adjusted for multiple comparisons using Tukey-Kramer post-hoc analyses). Significant adjusted p values <0.05 are italicized for comparisons not involving IVM, with the direction of significance noted directly to the right. Survival curves for each group are from 3 experimental replicates (other than only 2 replicates for initial solvent control experiments; 4 replicates for An. coluzzii AQ/DEAQ feeds). Data comparing survival curves for all blood only controls and IVM 1X feeds in An. gambiae versus An. coluzzii mosquitoes are also included at the bottom rows.
Fecundity for An. gambiae and An. coluzzii mosquitoes following ingestion of bloodmeal supplemented with solvents and tested antimalarials. For initial solvent control feeds in An. gambiae, data include the total number of eggs laid and eggs laid per mosquito (all fed mosquitoes monitored communally for egg laying, thus percent of mosquitoes laying eggs unknown (indicated by n/a value)) from two experimental replicates. For all further drug feeds, data includes the total eggs laid, number of eggs laid per mosquito, and percent of mosquitoes laying eggs (n=5 mosquitoes monitored in all egg laying conditions) following bloodmeal ingestion under each condition between experimental replicates (three replicates for An. gambiae and An. coluzzii AQ/DEAQ and SP feeds; 2 replicates for An. coluzzii PPQ feeds excluding further replicates from analyses where no mosquitoes laid eggs). Data for eggs per mosquito are reported as the mean (with Standard Deviation, and also with Standard Error of the Mean, respectively, separated by semi-colon). Data for total eggs laid and percent laying are reported as the mean (with Standard Deviation).
Raw egg laying data for lab-derived An. gambiae and field-derived An. coluzzii mosquitoes following ingestion of bloodmeal supplemented with tested antimalarials. Data are shown as the number of eggs laid per mosquito (n=5 mosquitoes monitored in individual egg laying conditions) following bloodmeal ingestion under each condition in three experimental replicates per group.
Drug detection comparisons in isolated An. gambiae mosquito compartments over time. Mosquitoes were fed 1 ɥg/ml or 250 ng/ml DEAQ to represent twice (2X) and half (1/2X) reported Cmax, respectively, or 50 ng/ml IVM to represent 1X reported Cmax. Controls included un-supplemented blood (Blood only), and solvent-supplemented blood (ACN:H2O 1:9 + 0.5% FA for DEAQ; DMSO for IVM). Mosquitoes were harvested 24hrs (all conditions) and 120hrs (Blood only, ACN:H2O 1:9 + 0.5% FA, DEAQ 2X, DEAQ 1/2X) post-feed, either collected whole or used for hemolymph collection and midgut dissection. For each condition, 5 of each mosquito sample type were pooled, with triplicates of pooled samples submitted to NorthEast BioLab (Hamden, CT) for LC-MS/MS analysis. Data is from 2 separate experiments. Results are listed for one-way ANOVA analyzing drug levels detected in whole mosquitoes, midguts, and hemolymph with pairwise comparisons of conditions of interest over time. Significant p values (p<0.05) are italicized.
Files and variables
File: Goheen_Impact_of_antimalarials_in_mosquitoes_Dryad.xlsx
Description: Because the data are not possible to universally curate in tabular format (in terms of tabulating individual mosquito data from all experiments, versus the analyses that were done by mosquito group in repeated experiments), this supplemental file consists of raw data numbers and relevant statistical analyses presented in table-like format across different Excel file tabs. Bolded words in the data description correspond to Excel tab labels, thus linking table descriptions to each Excel tab. In the Excel file, there are some empty cells present that visually help separate data from different experiments and/or conditions in the tables presented. Some merged cells also exist for ease of visualizing data tables presented here. Any cell that represents unavailable data (missing data points from experiments) is indicated by “null” value. Any cell that purposefully did not have data generated for a given experiment or analysis is indicated by “n/a.”
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