Data from: Morin hydrate reduces fertility and survival, delays development, and weakens lipid reserves in Aedes aegypti
Data files
Apr 02, 2025 version files 29.87 KB
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README.md
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Santos_et_al._2025_modify.xlsx
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Abstract
The Aedes aegypti mosquito is generally associated with arboviruses that cause yellow fever, dengue, zika and chikungunya. The most efficient way to control their populations is through application in breeding sites of highly toxic insecticides that can also impact non-target organisms and generate resistant populations. Therefore, the use of compounds is desirable. Morin hydrate has broad pharmacological applications based on its antioxidant potential, in addition to not having negative effects on mammals. Therefore, the objective of the present study was to investigate the effects of morin hydrate on A. aegypti survival, pupation rate, egg laying, triacylglycerol reserves, and expression of proteins related to lipid metabolism 24 h after exposure of larvae. For this, rearing media containing A. aegypti larvae with different concentrations of morin hydrate were formulated to evaluate the lethal concentration. Calculation of the expected lethal concentrations showed LC25 of 52.692 µM, LC40 of 111.121 µM, LC50 of 174.775 µM, LC75 of 575.083 µM, and LC90 of 1685.936 µM. Twenty-four hours after treatment with morin hydrate, surviving larvae were transferred to morin-free water with food, and their pupation rate and fertility were evaluated. We observed that an increase in the concentration of morin hydrate induced a dose-dependent reduction in survival, doubled pupation time in survivors, and reduced the number of eggs laid by treated females during the larval stage by approximately 30% at concentrations exceeding 100µM. From this, the impact of 24 h on the triacylglycerol (TAG) stock was evaluated, in addition to evaluating the expression of proteins involved in lipid metabolism. Larvae 24h after treatment with 100 µM morin showed a reduction in TAG reserves of approximately 17%, while at 175 µM, there was a reduction of more than 33% in stocks and at 500µM there was a reduction of 61%. Furthermore, the lipolytic proteins TAGL1 and HSL were upregulated, while the lipogenic proteins FAS1, DGAT1 and GPAT1 were downregulated. Insulin-like receptors were also downregulated, unlike AKHr, which was also upregulated. These data demonstrate that morin hydrate reduces the survival and fertility of A. aegypti by affecting its lipid metabolism. Morin hydrate did not exhibit toxicity toward non-target organisms, demonstrating interesting potential for the control of mosquito populations.
Dataset DOI: 10.5061/dryad.x0k6djhwh
Description of the data and file structure
Dataset Overview:
This dataset contains experimental data from Santos et al. (2025), testing the hypothesis that morin hydrate disrupts lipid metabolism in Aedes aegypti larvae, encompassing behavioral assays and molecular analyses. The dataset includes larval mortality data with LC25 (52.692 µM), LC50 (174.775 µM), and LC90 (1685.936 µM) values derived from dose-response curves. Survivors exhibited a 2-fold delay in pupation time and an approximately 30% reduction in egg-laying by females exposed to concentrations above 100 µM. Biochemical analyses revealed dose-dependent triacylglycerol depletion (reductions of 17%, 33%, and 61% at 100, 175, and 500 µM, respectively), which correlated with alterations in lipid metabolism gene expression, upregulation of lipases (Triacylglycerol Lipase 1 and Hormone-sensitive lipase), and downregulation of lipogenic enzymes (Fatty Acid Synthase 1, Diacylglycerol acyltransferase 1, and Glycerol-3-phosphate acyltransferase). Complementary data demonstrated the safety of this compound in mammals, with toxicological assays in mice showing no adverse effects.
Files and variables
Description of the Data and File Structure
The file was organized into thematic folders containing: (1) processed behavioral data (survival and pupation rates, egg laid and eclosion), (2) biochemical results (triacylglycerol values), and (3) molecular analyses (qPCR 2ΔΔCt values). Each manuscript figure has its corresponding dataset in sequentially named Excel files, accompanied by detailed metadata on the experimental conditions and sample sizes. Statistical analyses, including ANOVA and post-hoc tests, were exported from GraphPad Prism in formats compatible with other software packages. Raw spectrophotometry data (TAG assays) and qPCR cycle values are available in separate files for verification, while processed and normalized versions are provided for immediate result replication. The README file describes the complete structure and analysis protocols.
Files
Baseline Data
- Figure_1_Survival_Rate sheet
This file contains the survival rate of larvae treated with different concentrations of morin hydrate. L3 stage larvae were reared in solutions with different concentrations of morin, and the survival rate after 24 h was determined.
Number of variables: 1
Number of header rows: 1
Number of rows: 72
Variable list:
Morin hydrate concentration (µM): (numeric)
- Figure_2_Daily_mortality sheet
This file contains the daily mortality of larvae treated with different concentrations of morin hydrate for six days. L3 stage larvae were reared in solutions with different concentrations of morin, and the cumulative mortality rate was assessed over a 6-day period.
Number of variables: 2
Number of header rows: 1
Number of rows: 90
Variable list:
Post-treatment day: (numeric)
Morin hydrate concentration (µM): (numeric)
- Figure_2_Daily_Pupation sheet
This file contains the daily pupation of larvae treated with different concentrations of morin hydrate for six days. L3 stage larvae were reared in solutions with different concentrations of morin, and the cumulative pupation rate was assessed over a 6-day period.
Number of variables: 2
Number of header rows: 1
Number of rows: 90
Variable list:
Post-treatment day: (numeric) Post treatment days of morin hydrate concentrations.
Morin hydrate concentration (µM): (numeric)
- Figure_3_Egg_Laid sheet
This file contains the number of eggs laid by females that were treated with different concentrations of morin hydrate during the larval stage.
Number of variables: 1
Number of header rows: 1
Number of rows: 254
Variable list:
Morin hydrate concentration (µM): (numeric)
- Figure_4_Egg_eclosion_rate sheet
This file contains the number of eggs laid by adult females exposed to varying concentrations of morin hydrate during their larval stage, along with their eclosion rates.
Number of variables: 2
Number of header rows: 1
Number of rows: 104
Variable list:
Morin hydrate concentration (µM): (numeric)
Initial eggs: (numeric) The initial egg count was determined to establish the eclosion percentage based on the number of hatched larvae.
- Figure_5_Triacylglycerol sheet
This file contains the level of triacylglycerol in larvae that were treated with different concentrations of morin hydrate.
Number of variables: 1
Number of header rows: 1
Number of rows: 20
Variable list:
Morin hydrate concentration (µM): (numeric)
- Figure_6_Expression sheet
This file contains the relative expression levels of lipid metabolism proteins in larvae treated with different concentrations of morin hydrate. The data are expressed as the mean relative expression calculated (2^∆∆CT) and standard deviation.
Number of variables: 2
Number of header rows: 1
Number of rows: 80
Variable list:
Morin hydrate concentration (µM): (numeric)
Gene expression: (alphanumeric) Lipid metabolism proteins
Code/software
Microsoft® Excel® LTSC MSO (16.0.14332.21007)
GraphPad Prism 8.0.2
Mosquito Rearing
The study utilized Aedes aegypti (Rockefeller strain) larvae from a laboratory colony maintained under controlled conditions (27±1°C, 80±10% relative humidity, 12h light/dark cycle). Larvae were fed commercial cat food, while adults received a 10% sucrose solution. Blood meals were provided to 4-day-old females for egg production. All procedures followed ethical guidelines approved by CEUA/UFRRJ (protocol #006/2022).
Chemical Exposure
Third-instar larvae were exposed to morin hydrate (10-2500 µM) dissolved in 0.1% ethanol for 24 hours under fasting conditions. A control group received only the 0.1% ethanol vehicle. Larval density was maintained at 1 individual per mL of solution throughout the experiments.
Biological Assessments
Mortality was evaluated after 24-hour exposure across four replicates (10 larvae/concentration). For chronic effects, surviving larvae were transferred to clean water and monitored for pupation and adult emergence over six days. Adult fecundity was assessed by counting eggs laid 72 hours post-blood meal.
Biochemical Profiling
Triacylglycerol levels were quantified using a commercial enzymatic assay, with results normalized to total protein content determined by the Lowry method. Ten larvae per concentration were homogenized for analysis, with five biological replicates per treatment.
Gene Expression Analysis
RNA was extracted from larvae (TRIzol method) and reverse-transcribed following DNase treatment. qPCR assays evaluated expression of lipid metabolism genes (Triacylglycerol Lipase 1, Brummer lipase, Hormone-sensitive lipase, Fatty acid synthase, Diacylglycerol acyltransferase 1, Glycerol-3-phosphate acyltransferase, Adipokinetic hormone receptor, and Insulin like receptor) using SYBR Green chemistry. Reference genes (Actin/α-Tubulin) ensured data normalization, with amplification efficiency validated for all primers.
Statistical Approach
Dose-response relationships were analyzed via Probit regression. Treatment effects were evaluated by ANOVA with post-hoc tests (Prism 8.0.2), considering p<0.05 significant. All data are presented as mean ± SEM from at least three independent experiments.