Search Results (1,955)

Common methods for detecting Drosophila suzukii (spotted-wing drosophila, SWD) in fruit, such as microscopy, physical extraction, and incubation, are time-consuming and may underrepresent egg and first instar larvae counts, the smallest life stages of SWD. To address these limitations, we evaluated a quantitative real-time PCR (qPCR) protocol to detect and quantify SWD eggs using a linear model of the log-transformed ratio of eggs to sample volume (µL) in Tris buffer and fruit tissue. Compared to traditional approaches, this method reduces identification time from several weeks to approximately five hours. We observed a negative linear correlation between qPCR cycle threshold and egg concentration in both standard and fruit tissue samples, with similar model fits (R² = 0.7215 for field fruit tissue; R² = 0.874 for standard samples). This DNA-based protocol improves infestation detection speed and accuracy by enabling rapid, species-specific identification of D. suzukii in fruit tissue, addressing limitations of morphological identification of eggs and larvae. Further refinement for fruit tissue could enhance real-world applicability. Rapid detection may enable timely assessment of varietal resistance to SWD and support safer control strategies targeting early life stages, helping to prevent pest development and fruit degradation. Full article

The ecdysteroid receptor (EcR) plays a crucial role in insect development and metamorphosis, making it a promising target for the design of novel biorational compounds. This study investigated the cytotoxicity, as well as the EcR agonist and antagonist activities, of three synthetic molecules analogous to tebufenozide and extracts from nine plant species using the dipteran S2 cell line which originates from the insect model of the fruit fly Drosophila melanogaster. Cytotoxicity assays were performed to determine appropriate concentrations of the synthetic molecules and plant extracts for cell transfection. EcR agonist and antagonist activities were evaluated using 20-hydroxyecdysone (20E) as the control hormone. The synthetic molecules analogous to tebufenozide did not activate EcR in S2 cells. In contrast, the plant extract of Neoblechnum brasiliense, commonly known as Brazilian dwarf tree fern, exhibited significant antagonistic activity at 100 µM, reducing receptor activity by 92%, likely due to its phytosteroid content, and without inducing cytotoxic effects. These findings demonstrate that certain plant extracts, particularly N. brasiliense, act as effective EcR antagonists and may represent promising natural leads for the development of environmentally compatible biorational compounds to control economically important dipteran pests, such as fruit flies and mosquitoes. Full article

The current study investigates the modulatory effects of gallic acid (GA), 3-hydroxytyrosol (3-HT), and quercetin (QUE) on key cholinesterase enzymes using Drosophila melanogaster (fruit fly) head homogenates as a source of central cholinesterases following in vivo larval exposure. The choice of these plant phenolics was predicated on their cholinesterase (ChE) inhibitory effect reported recently by our group. The study utilized D. melanogaster larvae subjected to varying doses of GA, 3-HT, and QUE, subsequently evaluating enzymatic activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Galanthamine HBr was used as a positive control. All three phenolic compounds exhibited elevated ΔOD/min values for BChE inhibition compared to the negative control (ethanol). GA and QUE inhibited AChE, though with lower potency than galanthamine; at 1 mM, GA and QUE achieved 79.23% and 80.98% inhibition, respectively, compared to 98.34% for galanthamine. Interestingly, the effect of 3-HT on AChE was inversely related to the dose. The results indicate that GA and QUE modulate cholinesterase activity in vivo, consistent with our prior in vitro reports. This study also provides the first in vivo evidence of 3-HT’s ChE-modulating activity in Drosophila within a whole-organism model. Full article

The CRISPR–Cas9 system has significantly advanced genome editing but remains constrained by its requirement for specific protospacer adjacent motifs (PAMs). To overcome this limitation, PAM-relaxed nucleases, including the novel near-PAMless chimeric SpRYc, have been developed. Here, we evaluated SpRYc editing activity across multiple experimental systems, including human HEK293 and CEM-R5 cells, as well as Drosophila melanogaster S2 cells and embryos. In HEK293 cells, SpRYc exhibited broad PAM compatibility, enabling editing at non-canonical PAMs, albeit with reduced and variable efficiency at canonical NGG sites compared to SpCas9. This context dependency was more pronounced in CEM-R5 T cells, where SpRYc activity at endogenous CXCR4 and B2M loci was largely restricted to NGG PAMs. In contrast, unlike SpCas9, SpRYc displayed negligible genome-editing activity in Drosophila embryos in vivo. Notably, the transcriptional activator dSpRYc-VPR showed robust activity in Drosophila S2 cells at both canonical and non-canonical PAMs. Reduced chromatin occupancy of dSpRYc-VPR suggests a balance between expanded PAM recognition and DNA-binding stability, providing a mechanistic explanation for context-dependent performance of SpRYc. Overall, our results highlight that expanded targeting flexibility comes at the cost of variable efficiency, underscoring the need for extensive locus- and context-specific validation of PAM-relaxed genome-editing tools. Full article

The human lifespan has increased dramatically over the last few decades; however, reaching older age increases the risk of age-related diseases and ailments. To extend the healthspan, many have turned to supplements, including plant-based remedies used in traditional medicine, to promote healthy aging. One of these is Scutellaria lateriflora L. (S. lateriflora), native to North America, which has traditionally been used to treat anxiety, stress, and insomnia. However, clinical trials addressing its effects are very limited. Furthermore, plant material is intrinsically complex, and the preparation method affects the composition of extracts. We therefore used Drosophila to test whether S. lateriflora can confer resilience against age-related sleep and mobility deficits, using aqueous (SLAq) and ethanol extracts (SLE). Whereas both SLE and SLAq improved mobility, only SLE reduced sleep fragmentation in older males. By testing several flavonoids present in S. lateriflora, we found that the beneficial effects on mobility were mainly due to baicalin, whereas sleep was improved by a wogonin mix. Since neither the extracts nor the compounds extend the lifespan, this suggests that they improve neuronal health and function and do not generally slow down the aging process. This was supported by our finding that neuronal degeneration was reduced by S. lateriflora (SL) supplementation. Full article

Background/Objectives: Fungi produce a diverse array of metabolites, including various volatile organic compounds (VOCs) with known physiological functions and other biological activities. These metabolites hold significant potential for medical and industrial applications. Within the fungal domain, Penicillium species represent a particularly important group. Methods: This study characterized the VOC profiles of four Penicillium expansum strains (R11, R19, R21, and R27) and one Penicillium polonicum strain (RS1) using the solid-phase microextraction–gas chromatography–mass spectrometry technique. Results: The analysis revealed that the only compound in common among the five strains of Penicillium was phenyl ethanol. The high toxicity of P. polonicum RS1 to Drosophila larvae correlated with its diverse and abundant alkene production. Specifically, alkenes constituted 31.28% of its total VOCs, followed by alcohols at 29.13%. GC-MS analyses detected 22, 17, 22, and 18 specific VOCs from R11, R19, R21, and R27, respectively. Overall, alkenes dominated the R11 profile (17.03%), alcohols were most abundant in R19 (28.82%), and R21 showed the highest combined release of alcohols (23.2%) and alkenes (11.7%), while R27 produced a moderate abundance of alcohols (9.16%) and alkenes (4.19%). Among the P. expansum strains, R11, R21, and R27 exhibited substantially higher toxicity than R19 strain in our previous assessment; these findings are consistent with their respective VOC profiles. Conclusions: The distinct VOC compositions across Penicillium strains significantly influence their biological characteristics and ecological functions. These findings provide a basis for follow-up research into the mechanisms of fungal volatile-mediated toxicity and support the development of biocontrol strategies. Full article

Behavioral individuality, often termed animal personality, reflects consistent patterns of behavioral variability across individuals. In fruit flies (Drosophila melanogaster), pharmacological and dietary manipulations affecting neuromodulatory systems have been shown to alter behavior, but their effects on behavioral predictability remain incompletely understood. Here, we investigated whether developmental dietary exposure to tryptophan (a serotonin precursor) or escitalopram (a selective serotonin reuptake inhibitor, SSRI) is associated with changes in lateralized turning behavior. Flies were reared from larval stages on supplemented media and tested in a Y-maze assay to assess movement predictability. Flies exposed to escitalopram displayed significantly reduced behavioral variability compared to controls, indicated by a lower median absolute deviation (MAD) of turning behavior, whereas tryptophan supplementation did not significantly affect variability. Because both compounds were tested at a single dietary dose and serotonergic activity was not directly measured, these findings should be interpreted as dose-specific behavioral effects rather than evidence of altered serotonergic tone or mechanism. Our results demonstrate that chronic developmental exposure to escitalopram is associated with increased behavioral predictability in fruit flies, highlighting the utility of high-throughput behavioral assays for detecting subtle pharmacologically induced changes in individual variability. Future studies incorporating dose–response designs and physiological validation will be required to establish underlying mechanisms. Full article

Sensory systems allow the detection of external and internal cues essential for adaptive responses. Chemosensation exemplifies this integration, guiding feeding, mating, and toxin avoidance while also influencing physiological regulation. Across taxa, chemical detection relies on diverse receptor families, and emerging evidence reveals that transient receptor potential (TRP) channels—traditionally associated with phototransduction, thermosensation, and mechanotransduction—also mediate chemosensory functions. Studies in Drosophila melanogaster and vertebrates demonstrate that TRPs detect tastants, odorants, and internal chemical states, highlighting their evolutionary conservation and functional versatility. This review synthesizes current insights into the roles of TRP channels across four major domains: taste, smell, internal state, and central circuit modulation. Using D. melanogaster and mammalian systems as comparative frameworks, we highlight how TRP channels function as polymodal sensors, signal amplifiers, and modulators embedded within canonical receptor pathways rather than as standalone chemoreceptors. Recognizing these integrative functions not only expands our understanding of how organisms coordinate behavior with internal states but also points to TRP channels as potential targets for addressing chemosensory disorders and metabolic diseases. This framework highlights key directions for future research into TRP-mediated sensory and homeostatic regulation. Full article

In this study, we investigated the current and future potential distribution of three parasitoid species of Drosophila suzukii, which represent promising candidates for the biological control of this pest: Leptopilina japonica (Hymenoptera, Figitidae), Pachycrepoideus vindemmiae (Hymenoptera, Pteromalidae), and Trichopria drosophilae (Hymenoptera, Diapriidae). To this end, we employed Ecological Niche Modeling using the Random Forest algorithm and climatic data from WorldClim v. 2.1 under climate change scenarios (SSP2-4.5 and SSP5-8.5), analyzing the spatial overlap between the pest and its natural enemies. The results indicate that the parasitoids exhibit distinct geographic distributions, although most species show higher suitability for temperate regions of the Northern Hemisphere. Species such as T. drosophilae and L. japonica stand out for their broad distribution and high overlap with the pest, whereas P. vindemmiae and display more restrictive climatic ranges and lower control efficiency. With ongoing climate change, all parasitoids tend to migrate toward higher latitudes, with significant range contractions in tropical regions. Thus, our results demonstrate the usefulness of Ecological Niche Modeling in the selection of biological control agents by considering host-specific preferences and environmental requirements in the development of management strategies adapted to future scenarios. Full article

Live imaging has been instrumental in understanding cellular dynamics in Drosophila tissues, but technical limitations have prevented the long-term visualization of cell competition in adult brains. Here, we describe a simple ex vivo protocol that enables extended live imaging of adult Drosophila brains for up to 32 h. The method relies on non-supplemented Schneider’s Drosophila medium and hydrophobic interactions to maintain brain stability during imaging, eliminating the need for complex culture conditions or embedding procedures. We validate this approach by studying cell competition in the optic lobes following traumatic brain injury, where cell competition is expected to occur with a peak at 48 h after damage. We demonstrate the value of this method by visualizing the expression of the fitness checkpoint Azot in a loser cell and its subsequent elimination. This protocol offers a versatile platform for studying cell competition and other cellular processes requiring extended observation of the adult Drosophila brain. Full article

Lysosomal storage disorders (LSDs) are a group of rare inherited diseases caused by mutations in the genes encoding the proteins involved in normal lysosomal functions, leading to an accumulation of undegraded substrates within lysosomes. Among the most prominent clinical features are neurological impairment and neurodegeneration, arising from widespread cellular dysfunction. The development of powerful and reliable animal model systems that can in vivo recapitulate human LSD pathologies is critical for understanding disease mechanisms and advancing therapeutic strategies. In this study, we identified the Drosophila melanogaster orthologs of human LSD-related genes using the DIOPT tool and performed tissue-specific gene silencing along the brain–midgut axis via the use of GAL4/UAS and RNAi combined technologies. Transgenic fly models presented key features of human LSD pathologies, including significantly shortened lifespans and a progressive locomotor decline that serves as a measure for neuromuscular disintegration, following age- and sex-dependent patterns. These phenotypic parallels in pathology strongly support the functional relevance of the selected orthologs and underscore the value of Drosophila as a versatile in vivo model system for advanced LSD pathology research, offering state-of-the-art genetic tools for molecularly dissecting disease mechanisms and providing cutting-edge novel platforms for high-throughput genetic and/or pharmacological screening, moving towards development of new therapeutically beneficial drug-based regimens and mutant gene-rescue schemes. Full article

Several fruit fly species in the Drosophilidae and Tephritidae families are classified as invasive pests worldwide. However, limited information is available on the occurrence of those dipteran species, their interactions, and their host plants in the Pampa Biome of southern Brazil. Therefore, we investigated interactions between invasive fruit flies and host plants. After two years (2022–2023) of fruit surveys, 3802 fruits from 16 plant species were harvested in the Pampa Biome’s urban and rural areas. The collected fruits were conditioned in the laboratory. Emerged adult flies were identified and associated with the respective fruit species. The most abundant species detected were Zaprionus indianus (Gupta), followed by Zaprionus tuberculatus (Malloch), Drosophila suzukii (Matsumura), and Ceratitis capitata (Wiedemann). The invasive drosophilids D. suzukii, Z. indianus and Z. tuberculatus interacted with 10 host plant species each, whereas C. capitata was associated with seven host species. Zaprionus indianus showed the highest infestation rate on fruits collected from the ground, whereas D. suzukii had the highest infestation rate in fruits attached to plants. New host plants were identified for Z. tuberculatus and C. capitata. The findings of this study provide a valuable framework for more effective management and control strategies. Full article

Freshwater cladocerans such as Daphnia magna (D. magna) are keystone grazers whose hormone-regulated life history traits make them sensitive sentinels of endocrine-disrupting chemicals (EDCs). The organophosphate flame-retardant tris(2-butoxyethyl) phosphate (TBOEP) and perfluoroethylcyclohexane sulfonate (PFECHS) now co-occur at ng L⁻¹–µg L⁻¹ in surface waters, yet their chronic sub-lethal impacts on invertebrate endocrine networks remain unclear. We analysed two publicly available 21-day microarray datasets (TBOEP: GSE55132; PFECHS: GSE75607) using gene ontology enrichment, STRING protein interaction networks, Drosophila phenotype mapping, and KEGG (Kyoto Encyclopaedia of Genes and Genomes)-anchored frameworks to build putative adverse outcome pathways (AOPs) for D. magna. Differentially expressed genes were clustered into functional modules and hub nodes were ranked by degree and betweenness. TBOEP suppressed moulting and growth, altering 1157 genes enriched for metabolism and membrane processes; hubs VRK1, MIB2, and adenylosuccinate synthetase formed a muscle anatomical development sub-network. PFECHS down-regulated vitellogenin and shifted 879 genes dominated by oxidative-stress and glutathione-metabolism signatures; central nodes UBC9, eIF4A-III, Tra-2α, and HDAC1 linked meiotic-cycle, oogenesis, and cyclic-compound binding. Despite chemical dissimilarity, both compounds converged on Wnt-signalling nodes—TBOEP via presenilin-1, and PFECHS via CK1ε/CK2—thereby reducing TCF/LEF-dependent transcription. Predicted outcomes include impaired oocyte maturation, reduced fecundity, and stunted body size, consistent with observed decreases in length and vitellogenin protein. Our network analysis, based on high-dose, sub-lethal exposures used in the underlying microarray studies, indicates that TBOEP- and PFECHS-induced perturbations can destabilise endocrine, developmental, and metabolic pathways in D. magna without overt lethality, and highlights Wnt-centred key events and hub genes as candidate biomarkers to be evaluated in future low-dose studies that use environmentally realistic exposure scenarios. Hub genes and Wnt-mediated key events emerge as sensitive biomarkers for monitoring mixed EDC exposure. Full article

Alzheimer’s Disease (AD) is a neurodegenerative disorder characterised by Amyloid-beta 42 (Aβ42) plaque accumulation and cognitive decline, with current treatments focused on symptomatic relief. Emerging therapeutics, such as dietary interventions, can modulate cognitive decline and delay AD progression. Our previous work in Drosophila melanogaster identified cell competition as a key mechanism that eliminates unfit neurons in an AD model, improving locomotion by removing the unfit neurons expressing flower^LoseB and ahuizotl (azot). Here, we explored how diet influences azot-dependent cell competition and locomotion in the AD model. Flies were fed with either a yeast-based diet (YBD) or a synthetic (SAA) diet for up to 28 days. In contrast to YBD, SAA delayed cell competition activation until day 21, coinciding with locomotion improvement and delayed Aβ formation. The overexpression of the human Flower (hFWE) isoforms in a Drosophila neuronal context revealed functional conservation: hFWE1 acted as the sole loser isoform, and hFWE2 as a winner isoform. With the YBD, forcing cell competition by expressing hFWE2 in the AD model led to an accumulation of unfit cells and promoted worse locomotion phenotypes over time compared to with the SAA diet. Our data highlights the complex interaction between diet, cell competition, and Aβ toxicity, offering new therapeutic insights. Full article

Research into longevity and aging involves comparing the size of cohorts at certain points on survival curves. However, this analysis is oversimplified because it provides limited information about the sample structure and the distribution of lifespan as a trait. Here, we introduce a method for estimating lifespan across the entire data range using distribution analysis. More specifically, we propose dividing the lifespan series into intervals, obtaining the frequencies of phenotypes by lifespan within the sample, followed by distribution analysis using the normality criterion. Additionally, to visualize the differences, we propose describing the resulting distributions formally using the normal distribution function and the β-distribution function. We demonstrate that the proposed methodology enables to extract additional information from survival data, providing new insights into the processes that occur in populations in response to genetic interventions and shedding light on their impact on ontogenesis. In particular, we observed that the lifespan distribution in Drosophila may not meet the normality criterion and may take different shapes depending on the line’s genotype or in response to genetic interventions. The proposed approach adds a new layer of information to studies of longevity and aging and expands the toolkit of methods used to analyze survival data. Full article