DOI: 10.5061/dryad.x0k6djhwh
title: Data from: Morin hydrate reduces fertility and survival, delays development, and weakens lipid reserves in Aedes aegypti

description: 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.
description: Mosquito Rearing The study utilized
 <em>Aedes aegypti</em> (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&lt;0.05 significant. All data are presented as mean ± SEM from at
 least three independent experiments.
description: # Morin hydrate reduces fertility and survival, delays development, and
 weakens lipid reserves in Aedes aegypti Dataset DOI:
 [10.5061/dryad.x0k6djhwh](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