RESUMEN
This study explores the green synthesis of silver nanoparticles (AgNPs) using a methanolic extract of fermented pollen from Tetragonisca angustula, a species of stingless bees. The AgNPs exhibit spherical morphology, low charge values, and suspension stability, with their unique composition attributed to elements from the pollen extract. Antioxidant assays show comparable activity between the pollen extract and AgNPs, emphasizing the retention of antioxidant effects. The synthesized AgNPs demonstrate antimicrobial activity against multidrug-resistant bacteria, highlighting their potential in combating bacterial resistance. The AgNPs exhibit no toxic effects on Drosophila melanogaster and even enhance the hatching rate of eggs. The study underscores the innovative use of stingless bee pollen extract in green synthesis, offering insights into the varied applications of AgNPs in biomedicine.
RESUMEN
Stingless bees are a diverse group with a relevant role in pollinating native species. Its diet is rich in carbohydrates and proteins, by collecting pollen and nectar supplies the development of its offspring. Fermentation of these products is associated with microorganisms in the colony. However, the composition of microorganisms that comprise this microbiome and its fundamental role in colony development is still unclear. To characterize the colonizing microorganisms of larval food in the brood cells of stingless bees Frieseomelitta varia, Melipona quadrifasciata, Melipona scutellaris, and Tetragonisca angustula, we have utilized molecular and culture-based techniques. Bacteria of the phyla Firmicutes, Proteobacteria, Actinobacteria, and fungi of the phyla Ascomycota, Basidiomycota, Mucoromycota, and Mortierellomycota were found. Diversity analysis showed that F. varia had a greater diversity of bacteria in its microbiota, and T. angustula had a greater diversity of fungi. The isolation technique allowed the identification of 189 bacteria and 75 fungi. In summary, this research showed bacteria and fungi associated with the species F. varia, M. quadrifasciata, M. scutellaris, and T. angustula, which may play an essential role in the survival of these organisms. Besides that, a biobank with bacteria and fungus isolates from LF of Brazilian stingless bees was created, which can be used for different studies and the prospection of biotechnology compounds.
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Hongos , Levaduras , Abejas , Animales , Larva , Brasil , Hongos/genética , Bacterias/genéticaRESUMEN
Morphological characteristics of pequi fruits (Caryocar brasiliense) allows us to obtain information for the conservation, breeding, and detect genetic variability. Thus, the aims of this research were to characterize the genetic diversity of Caryocar brasiliense that produce fruits with and without thorn at the endocarp; determine the important characters in the study of genetic diversity, and to estimate phenotypic and genotypic correlations. 80 fruits with thorns and 90 fruits without thorns were evaluated. Data were used for analysis: Euclidian distance as a measure of dissimilarity and the Tocher method for delimitation of groups; Principal Component Analysis, and genotypic and phenotypic correlations. Genetic dissimilarity of C. brasiliense ranged from 0.066 (B6-D6) to 0.908 (A14-B3). Tocher method allowed dividing the matrices into three groups, the dendrogram into five groups, and the principal components allowed the distribution in 8 groups. We observed 15 significant correlations for characters of fruit with thorns and 31 correlations for characters of fruit without thorns. The weight of the fruit (with thorns) and the pyrene yield (without thorns) are relevant correlations for production. The groupings generated allowed the classification of the matrices into distinct sets. Allowed the identification of potential producers plants that could be used in breeding programs.
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Ericales , Frutas , Frutas/genética , Variación Genética/genética , FitomejoramientoRESUMEN
Alzheimer's disease (AD) is characterized by progressive, irreversible loss of memory and cognitive function. Drosophila melanogaster and other animal models are used to study several diseases, in order to elucidate unknown mechanisms and develop potential therapies. Molecular studies require biological samples and, for neuropathologies such as AD biopsy of the human brain, are invasive and potentially damaging. The solution is to use animal models, such as D. melanogaster, which is a model organism that can replace mammalian organisms in such studies. In this study, we evaluated the climbing ability and differential gene expression during AD progression due to the amylodoigenic pathway using RNA-seq, and we performed an in silico analysis of a fruit fly AD-like GFP (Green Fluorescent Protein) model with GFP expression in the pan-neural elav driver. A total of 1388 genes were differentially expressed in all analyzed groups. The main pathways related to those Differentially Expressed Genes (DEGs) during aging and AD progression were evaluated using the fly genes and human orthologs, in order to link genomic information to higher-order functional information with gene pathway mapping. We identified pathways present in all analyzed groups, such as metabolic pathways, ribosomal pathways, proteasome pathways and immune system pathways. Some of the genes were validated by qPCR. Knockdown of CG17754 gene by RNAi promoted degeneration in the fly eye, validating these findings in vivo. The identification of similarities in molecular pathways between the transgenic fly AD-like GFP model and mammals related to AD provides new insights into the use of this fly in screening novel anti-AD drugs.
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Enfermedad de Alzheimer , Drosophila melanogaster , Enfermedad de Alzheimer/metabolismo , Animales , Encéfalo/metabolismo , Modelos Animales de Enfermedad , Drosophila melanogaster/genética , Expresión Génica , Mamíferos , RNA-SeqRESUMEN
In bees from genus Melipona, differential feeding is not enough to fully explain female polyphenism. In these bees, there is a hypothesis that in addition to the environmental component (food), a genetic component is also involved in caste differentiation. This mechanism has not yet been fully elucidated and may involve epigenetic and metabolic regulation. Here, we verified that the genes encoding histone deacetylases HDAC1 and HDAC4 and histone acetyltransferase KAT2A were expressed at all stages of Melipona scutellaris, with fluctuations between developmental stages and castes. In larvae, the HDAC genes showed the same profile of Juvenile Hormone titers-previous reported-whereas the HAT gene exhibited the opposite profile. We also investigated the larvae and larval food metabolomes, but we did not identify the putative queen-fate inducing compounds, geraniol and 10-hydroxy-2E-decenoic acid (10HDA). Finally, we demonstrated that the histone deacetylase inhibitor 10HDA-the major lipid component of royal jelly and hence a putative regulator of honeybee caste differentiation-was unable to promote differentiation in queens in Melipona scutellaris. Our results suggest that epigenetic and hormonal regulations may act synergistically to drive caste differentiation in Melipona and that 10HDA is not a caste-differentiation factor in Melipona scutellaris.
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Abejas/fisiología , Conducta Alimentaria/fisiología , Regulación del Desarrollo de la Expresión Génica , Jerarquia Social , Monoterpenos Acíclicos/metabolismo , Animales , Epigénesis Genética , Ácidos Grasos/metabolismo , Ácidos Grasos Monoinsaturados/metabolismo , Femenino , Histona Acetiltransferasas/genética , Histona Acetiltransferasas/metabolismo , Histona Desacetilasas/genética , Histona Desacetilasas/metabolismo , Proteínas de Insectos/antagonistas & inhibidores , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Hormonas Juveniles/metabolismoRESUMEN
Vitamin D3 (VD3) deficiency increases DNA damage, while supplementation may exert a pro-oxidant activity, prevent viral infections and formation of tumors. The aim of this study was to investigate the mutagenicity and carcinogenicity of VD3 alone or in combination with doxorubicin (DXR) using the Somatic Mutation and Recombination Test and the Epithelial Tumor Test, both in Drosophila melanogaster. For better understanding of the molecular interactions of VD3 and receptors, in silico analysis were performed with molecular docking associated with molecular dynamics. Findings revealed that VD3 alone did not increase the frequency of mutant spots, but reduced the frequency of mutant spots when co-administered with DXR. In addition, VD3 did not alter the recombinogenic effect of DXR in both ST and HB crosses. VD3 alone did not increase the total frequency of tumor, but significantly reduced the total frequency of tumor when co-administered with DXR. Molecular modeling and molecular dynamics between calcitriol and Ecdysone Receptor (EcR) showed a stable interaction, indicating the possibility of signal transduction between VD3 and EcR. In conclusion, under these experimental conditions, VD3 has modulatory effects on the mutagenicity and carcinogenicity induced by DXR in somatic cells of D. melanogaster and exhibited satisfactory interactions with the EcR.
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Carcinogénesis/efectos de los fármacos , Colecalciferol/farmacología , Doxorrubicina/toxicidad , Drosophila melanogaster/efectos de los fármacos , Animales , Antibióticos Antineoplásicos/toxicidad , Hormonas y Agentes Reguladores de Calcio/farmacología , Drosophila melanogaster/genética , Femenino , Masculino , Modelos Moleculares , Simulación de Dinámica Molecular , Estructura Molecular , Mutación/efectos de los fármacos , Conformación Proteica , Receptores de Calcitriol/química , Receptores de Calcitriol/metabolismo , Recombinación GenéticaRESUMEN
A biological assessment of environmental quality was performed using the tropical plant species Tradescantia pallida (Rose) D.R. Hunt. var. purpurea exposed to different levels of air contamination in urban intersections with high volume of vehicle traffic. Air quality (average daily levels of particulate material in the PM1, 2.5, 10 fractions) and traffic volume in crossing intersections were monitored for 30 days before the collection of plants. Frequency of micronuclei and pollen abortivity in inflorescences collected at different intersections with gradual levels of traffic volume were evaluated as biomarkers of genotoxicity. In addition, the concentrations of bioaccumulated heavy metals in the leaves of the collected plants were also investigated. The proposed biological assessment model found a positive association between the environmental variables (traffic volume; concentration of particulate material) and biological effects (leaf concentration of Cr and Cd; micronucleus frequencies and pollen abortivity).
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Contaminantes Atmosféricos/toxicidad , Daño del ADN , Tradescantia/genética , Contaminación por Tráfico Vehicular/efectos adversos , Salud Urbana , Contaminación del Aire/análisis , Monitoreo del Ambiente/métodos , Metales Pesados/farmacología , Pruebas de Micronúcleos , Pruebas de Mutagenicidad , Hojas de la Planta/química , Hojas de la Planta/efectos de los fármacos , Polen/efectos de los fármacosRESUMEN
Water pollution and the increase in genotoxic consequences in aquatic environments are well documented indicating the necessity and importance of biomonitoring programs. The objective of the present study was to determine the environmental quality of water resources and genotoxic potential of materials present within water samples obtained from the Perdizes River and the Mumbuca Stream, located in a region of discharge of wastewater treatment effluents using Tradescantia micronucleus assay (Trad - MCN). Water samples were collected from different locations up and downstream of the wastewater treatment plant during rainy season and subsequently submitted to physico-chemical analysis and Trad-MCN bioassay. The spatial distribution of the physico-chemical parameters assessed suggested that discharges of wastewater treatment effluents reduced water quality at all sites examined. Further, exposure to wastewater treatment effluents produced genotoxic effects on tetrads of Tradescantia pallida. These results reinforce the sensitivity of the Trad-MCN bioassay and its potential application in water quality monitoring programs concomitant with physicochemical evaluation.
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Residuos Industriales/efectos adversos , Pruebas de Micronúcleos , Tradescantia/efectos de los fármacos , Aguas Residuales/toxicidad , Contaminantes Químicos del Agua/toxicidad , Tradescantia/genéticaRESUMEN
Antiparasitic substances are chemicals used to control or kill endoparasites and ectoparasites. Based on the premise that Ivermectin (IVM) and Amoxicillin (AMX) are commonly considered in parasitic control in mammals, the present study aimed to evaluate the carcinogenic and genotoxic potential of different concentrations of IVM and AMX through the detection of epithelial tumor test in Drosophila melanogaster. Third-instar larvae descending from the cross between wts/TM3, Sb1 females and mwh/mwh males were treated with different concentrations of IVM (2.9, 5.8, 11.6 and 23.2 x 10-17 mM) or AMX (1.37, 2.74, 5.48 and 10.9 x 10-16mM). The results revealed that IVM increased the frequency of epithelial tumor in D. melanogaster considering all evaluated concentrations, while AMX showed no carcinogenic effect. Furthermore, the Micronucleus (MN) test in Tradescantia pallida was used to evaluate the genotoxic effect of IVM and AMX. T. pallida individuals were exposed for 8 hours at different concentrations of IVM (5.71, 11.42, 22.84 and 45.68 x 10-5mM) or AMX (5.13, 10.26, 20.52 and 41.05 x 10-3mM). Findings showed an increase in the frequency of micronuclei in T. pallida treated with 11.42, 22.84 and 45.68 x 10-5mM of IVM. We conclude that chronic exposure to IVM is directly associated with events resulting from genetic instability (genotoxicity and carcinogenicity). On the other hand, AMX was neither carcinogenic nor genotoxic for D. melanogaster and T. pallida.
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Amoxicilina/toxicidad , Antiparasitarios/toxicidad , Carcinógenos/toxicidad , Ivermectina/toxicidad , Mutágenos/toxicidad , Animales , Carcinoma/inducido químicamente , Daño del ADN , Drosophila melanogaster/efectos de los fármacos , Drosophila melanogaster/genética , Femenino , Larva/efectos de los fármacos , Larva/genética , Masculino , Pruebas de Mutagenicidad , Tradescantia/efectos de los fármacos , Tradescantia/genéticaRESUMEN
Neonicotinoids and phenylpyrazoles are classes of neurotoxic insecticides which are able to bind at different ligand sites of neural receptors, leading to the deregulation of insect neural activity and hence resulting in death. The misuse or indiscriminate use of these chemicals is directly associated with several toxicological effects in biota and at different trophic levels. Based on this premise, the aim of the present study was to evaluate and compare the genotoxic capacity of different concentrations of thiamethoxam (TMX), acetamiprid (ACP), imidacloprid (IMI) and fipronil (FP) through the Micronucleus Test in Tradescantia pallida (Trad-MCN). After acclimatization (24â¯h), T. pallida stems were treated with stablished concentrations of TMX, ACP, IMI and FP for 8â¯h. Then, the stems of the model organism were submitted to a recovery phase (24â¯h). The young inflorescences were harvested and fixed in Carnoy solution and, after 24â¯h, were conserved in ethanol 70% until the analyzes. The obtained anthers were macerated on slides for microscopy, stained with acetic carmine dye and covered with coverslips before analysis by light microscopy. Considering the insecticides, the micronuclei (MN) frequency in plants treated at concentrations of 0.2 and 0.4â¯gâ¯L-1 for TMX, 0.2; 0.4 and 0.8â¯gâ¯L-1 for ACP, 0.1; 0.2; 0.4; 0.8 and 1.6â¯gâ¯L-1 for IMI and 0.2; 0.4; 0.8 and 1.6â¯gâ¯L-1 for FP differed statistically (pâ¯<â¯0.05, Tukey) from the MN frequency of the negative control. All chemicals evaluated revealed genotoxic activity in T. pallida at the highest concentrations.
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Insecticidas/toxicidad , Pruebas de Micronúcleos , Tradescantia/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Pruebas de Mutagenicidad , Neonicotinoides/toxicidad , Nitrocompuestos/toxicidad , PirazolesRESUMEN
Spinosad (SPN) is a naturally-occurring insecticide obtained from the fermentation process of the actinomycete Saccharopolyspora spinosa. Owing to the larvicidal action, the compound has been used in the control of Aedes aegypti. As a new insecticide commercially available in the market, few data are reported on genotoxic effects in non-target organisms. The objective of the present study was to evaluate the mutagenic effect of SPN through the Micronucleus Test in Tradescantia pallida (Trad-MCN) and using the mutation and somatic recombination test in Drosophila melanogaster (SMART). At the Trad-MCN, after acclimatization (24â¯h), T. pallida stems were submitted to chronic treatment with SPN at concentrations of 0.156; 0.312; 0.625; 1.25 and 2.5â¯g/L solution for 24â¯h, followed by a recovery period. In SMART, considering the third stage larvae, offspring resulting from the ST and HB crossing were placed on chronic treatment (48â¯h) with 0.039; 0.078 and 0.156⯵g/mL of SPN solution. No mutagenic effect was observed at any of the evaluated concentrations in SMART. Additionally, SPN is more toxic after metabolism via CYP6A2 (cytochrome P450) in D. melanogaster. However, SPN at the concentrations of 0.625; 1.25 and 2.5â¯g/L was able to induce high frequency of micronuclei in T. pallida. Under the experimental conditions of T. pallida in the present study, SPN caused genotoxic activity.
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Drosophila melanogaster/efectos de los fármacos , Macrólidos/toxicidad , Pruebas de Mutagenicidad/métodos , Tradescantia/efectos de los fármacos , Animales , Proteínas de Drosophila , Drosophila melanogaster/genética , Combinación de Medicamentos , Femenino , Insecticidas/toxicidad , Larva/efectos de los fármacos , Masculino , Pruebas de Micronúcleos , Mutágenos/toxicidad , Tradescantia/genéticaRESUMEN
Changes in microRNAs (miRNAs) expression have been described in major depressive disorder in young and middle-aged adults. However, no study has evaluated miRNA expression in older adults with major depression (or late-life depression [LLD]). Our primary aim was to evaluate the expression of miRNAs in subjects with LLD. We first evaluated the miRNA expression using next-generation sequencing (NGS) and then we validated the miRNAs found in NGS in an independent sample of LLD patients, using RT-qPCR. Drosophila melanogaster model was used to evaluate the impact of changes in miRNA expression on behavior. NGS analysis showed that hsa-miR-184 (log2foldchangeâ¯=â¯-4.21, pâ¯=â¯1.2â¯×â¯10-03) and hsa-miR-1-3p (log2foldchangeâ¯=â¯-3.45, pâ¯=â¯1.3â¯×â¯10-02) were significantly downregulated in LLD compared to the control group. RT-qPCR validated the downregulation of hsa-miR-184 (pâ¯<â¯0.001), but not for the hsa-miR-1-3p. The knockout flies of the ortholog of hsa-miR-184 showed significantly reduced locomotor activity at 21-24â¯d.p.e (pâ¯=â¯0.04) and worse memory retention at 21-24â¯d.p.e (24h post-stimulus, pâ¯=â¯0.02) compared to control flies. Our results demonstrated that subjects with LLD have significant downregulation of hsa-miR-184. Moreover, the knockout of hsa-miR-184 in flies lead to depressive-like behaviors, being more pronounce in older flies.
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Envejecimiento/genética , Conducta Animal , Disfunción Cognitiva/genética , Trastorno Depresivo Mayor/genética , Locomoción , MicroARNs/genética , Retención en Psicología , Factores de Edad , Anciano , Animales , Animales Modificados Genéticamente , Modelos Animales de Enfermedad , Regulación hacia Abajo , Proteínas de Drosophila , Drosophila melanogaster , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Locomoción/genética , Masculino , Persona de Mediana Edad , Análisis de Secuencia de ARN , Investigación Biomédica TraslacionalRESUMEN
Melipona scutellaris Latreille, 1811 (Hymenoptera, Apidae) is a pollinator of various native and cultivated plants. Because of the expansion of agriculture and the need to ensure pest control, the use of insecticides such as fipronil (FP) has increased. This study aimed to evaluate the effects of sublethal doses of FP insecticide on M. scutellaris at different time intervals (6, 12, and 24â¯h) after exposure, via individually analyzed behavioral biomarkers (locomotor activity, behavioral change) as well as the effect of FP on different brain structures of bees (mushroom bodies, antennal cells, and optic cells), using sub-individual cell biomarkers (heterochromatin dispersion, total nuclear and heterochromatic volume). Forager bees were collected when they were returning to the nest and were exposed to three different concentrations of FP (0.40, 0.040, and 0.0040â¯ng a.i/bee) by topical application. The results revealed a reduction in the mean velocity, lethargy, motor difficulty, paralysis, and hyperexcitation in all groups of bees treated with FP. A modification of the heterochromatic dispersion pattern and changes in the total volume of the nucleus and heterochromatin were also observed in the mushroom bodies (6, 12, and 24â¯h of exposure) and antennal lobes (6 and 12â¯h) of bees exposed to 0.0040â¯ng a.i/bee (LD50/100). FP is toxic to M. scutellaris and impairs the essential functions required for the foraging activity.
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Ecotoxicología/métodos , Insecticidas/efectos adversos , Animales , Abejas , BrasilRESUMEN
Physico-chemical and toxicological analyses are of fundamental importance to determine water quality. The objectives of the present study were to evaluate the toxicity, mutagenicity and carcinogenicity of samples from the Mumbuca Stream and the Perdizes River, through both SMART and the wts test, respectively, in somatic cells of Drosophila melanogaster and to quantify the amount of heavy metals and other pollutants, which are indicative of environmental quality. Water samples were collected (M1, M2, P1, P2 and MP) and submitted to physico-chemical analysis, calculating the water quality index for each sampling site. In order to evaluate the toxicity, mutagenicity and carcinogenicity of the samples, third instar larvae descended from the crossing between virgin female wts/TM3, sb1 and mwh/mwh males (wts test) and ST and HB (SMART) crosses were treated with samples from P1, P2, M1, M2 and MP sites. The physico-chemical analysis and the biological assay allowed us to conclude that undetected values for heavy metals and the low frequency of mutant spots (SMART) and epithelial tumor (wts) in treated flies from the Mumbuca Stream and Perdizes River may be due to the reduction of ceramic activities in the municipality. The physico-chemical analyzes identified altered the environmental quality parameters, which directly influenced the survival of D. melanogaster treated with samples of M2 and MP, which according to the WQI were classified as regular and poor environmental quality, respectively. The altered parameters may be due to clandestine domestic sewage sent downstream of the effluent.
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Monitoreo del Ambiente/métodos , Mutágenos/toxicidad , Ríos/química , Aguas del Alcantarillado/análisis , Calidad del Agua , Animales , Drosophila melanogaster/citología , Drosophila melanogaster/efectos de los fármacos , Femenino , Larva/efectos de los fármacos , Masculino , Metales Pesados/análisisRESUMEN
Thiamethoxam (TMX) belongs to a class of neuro-active insecticides referred as neonicotinoids, while actara® (AC) is one of the most popular TMX-based products in Brazil. The aim of this study was to evaluate the mutagenic, recombinogenic and carcinogenic potential of TMX and AC insecticides. The mutagenic and recombinogenic effect of TMX and AC were evaluated in vivo by the Somatic Mutation and Recombination Test (SMART) while carcinogenic effects were evaluated through the Test for Detection of Epithelial Tumor Clones (wts test), both in somatic cells of Drosophila melanogaster. In the SMART, third instar larvae from standard (ST) and high bioactivation (HB) crosses were treated with different concentrations of TMX and AC (2.4; 4.8; 9.7 × 10-4 mM and 1.9 × 10-3 mM). The results revealed mutagenic effects at the highest concentrations tested in the HB cross. In the test for the detection of epithelial tumor, third instar larvae resulting from the cross between wts/TM3, Sb1 virgin females and mwh/mwh males were treated with the same concentrations of TMX and AC used in the SMART. No carcinogenic effect was observed at any of the concentrations tested. In this work, the inhibition of the mechanism of repair by homologous recombination was observed in flies exposed to 9.7 × 10-4 and 1.9 × 10-3 mM of AC. In conclusion, TMX and AC demonstrated to be a promutagen in the highest concentrations tested.
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Drosophila melanogaster/efectos de los fármacos , Neonicotinoides/farmacología , Nitrocompuestos/farmacología , Oxazinas/farmacología , Tiazoles/farmacología , Animales , Brasil , Carcinogénesis/efectos de los fármacos , Drosophila melanogaster/citología , Femenino , Insecticidas/farmacología , Masculino , Mutagénesis/efectos de los fármacos , Pruebas de Mutagenicidad/métodos , Recombinación Genética/efectos de los fármacos , TiametoxamRESUMEN
In social insects, juvenile hormone (JH) has acquired novel functions related to caste determination and division of labor among workers, and this is best evidenced in the honey bee. In contrast to honey bees, stingless bees are a much more diverse group of highly eusocial bees, and the genus Melipona has long called special attention due to a proposed genetic mechanism of caste determination. Here, we examined methyl farnesoate epoxidase (mfe) gene expression, encoding an enzyme relevant for the final step in JH biosynthesis, and measured the hemolymph JH titers for all life cycle stages of Melipona scutellaris queens and workers. We confirmed that mfe is exclusively expressed in the corpora allata. The JH titer is high in the second larval instar, drops in the third, and rises again as the larvae enter metamorphosis. During the pupal stage, mfe expression is initialy elevated, but then gradually drops to low levels before adult emergence. No variation was, however, seen in the JH titer. In adult virgin queens, mfe expression and the JH titer are significantly elevated, possibly associated with their reproductive potential. For workers we found that JH titers are lower in foragers than in nurse bees, while mfe expression did not differ. Stingless bees are, thus, distinct from honey bee workers, suggesting that they have maintained the ancestral gonadotropic function for JH. Hence, the physiological circuitries underlying a highly eusocial life style may be variable, even within a monophyletic clade such as the corbiculate bees.
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Abejas/genética , Proteínas de Insectos/genética , Hormonas Juveniles/metabolismo , Oxigenasas/genética , Animales , Abejas/crecimiento & desarrollo , Abejas/metabolismo , Femenino , Proteínas de Insectos/metabolismo , Larva/genética , Larva/metabolismo , Masculino , Oxigenasas/metabolismo , Filogenia , Pupa/genética , Pupa/metabolismo , Análisis de Secuencia de ADNRESUMEN
There is growing evidence in the literature suggesting that caste differentiation in the stingless bee, Melipona scutellaris, and other bees in the genus Melipona, is triggered by environmental signals, particularly a primer pheromone. With the proper amount of food and a chemical stimulus, 25% of females emerge as queens, in agreement with a long-standing "two loci/two alleles model" proposed in the 1950s. We surmised that these larvae must be equipped with an olfactory system for reception of these chemical signals. Here we describe for the first time the diversity of antennal sensilla in adults and the morphology of larvae of M. scutellaris. Having found evidence for putative olfactory sensilla in larvae, we next asked whether olfactory proteins were expressed in larvae. Since the molecular basis of M. scutellaris is still unknown, we cloned olfactory genes encoding chemosensory proteins (CSP) and odorant-binding proteins (OBPs) using M. scutellaris cDNA template and primers designed on the basis CSPs and OBPs previously reported from the European honeybee, Apis mellifera. We cloned two CSP and two OBP genes and then attempted to express the proteins encoded by these genes. With a recombinant OBP, MscuOBP8, and a combinatorial single-chain variable fragment antibody library, we generated anti-MscuOBP8 monoclonal antibody. By immunohistochemistry we demonstrated that the anti-MscuOBP8 binds specifically to the MscuOBP8. Next, we found evidence that MscuOBP8 is expressed in M. scutellaris larvae and it is located in the mandibular region, thus further supporting the hypothesis of olfactory function in immature stages. Lastly, molecular modeling suggests that MscuOBP8 may function as a carrier of primer pheromones or other ligands.
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Abejas/genética , Proteínas de Insectos/genética , Larva/genética , Percepción Olfatoria/genética , Receptores Odorantes/genética , Sensilos/metabolismo , Animales , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/metabolismo , Abejas/crecimiento & desarrollo , Abejas/metabolismo , Abejas/ultraestructura , Proteínas Portadoras/química , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Clonación Molecular , Femenino , Regulación del Desarrollo de la Expresión Génica , Inmunohistoquímica , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Larva/crecimiento & desarrollo , Larva/metabolismo , Larva/ultraestructura , Modelos Moleculares , Biblioteca de Péptidos , Feromonas/química , Feromonas/genética , Feromonas/metabolismo , Unión Proteica , Estructura Secundaria de Proteína , Receptores Odorantes/química , Receptores Odorantes/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Sensilos/crecimiento & desarrollo , Sensilos/ultraestructura , Anticuerpos de Cadena Única/química , Anticuerpos de Cadena Única/metabolismoRESUMEN
Stingless bees of the genus Melipona, have long been considered an enigmatic case among social insects for their mode of caste determination, where in addition to larval food type and quantity, the genotype also has a saying, as proposed over 50 years ago by Warwick E. Kerr. Several attempts have since tried to test his Mendelian two-loci/two-alleles segregation hypothesis, but only recently a single gene crucial for sex determination in bees was evidenced to be sex-specifically spliced and also caste-specifically expressed in a Melipona species. Since alternative splicing is frequently associated with epigenetic marks, and the epigenetic status plays a major role in setting the caste phenotype in the honey bee, we investigated here epigenetic chromatin modification in the stingless bee Melipona scutellaris. We used an ELISA-based methodology to quantify global methylation status and western blot assays to reveal histone modifications. The results evidenced DNA methylation/demethylation events in larvae and pupae, and significant differences in histone methylation and phosphorylation between newly emerged adult queens and workers. The epigenetic dynamics seen in this stingless bee species represent a new facet in the caste determination process in Melipona bees and suggest a possible mechanism that is likely to link a genotype component to the larval diet and adult social behavior of these bees.
RESUMEN
Abstract Stingless bees of the genus Melipona, have long been considered an enigmatic case among social insects for their mode of caste determination, where in addition to larval food type and quantity, the genotype also has a saying, as proposed over 50 years ago by Warwick E. Kerr. Several attempts have since tried to test his Mendelian two-loci/two-alleles segregation hypothesis, but only recently a single gene crucial for sex determination in bees was evidenced to be sex-specifically spliced and also caste-specifically expressed in a Melipona species. Since alternative splicing is frequently associated with epigenetic marks, and the epigenetic status plays a major role in setting the caste phenotype in the honey bee, we investigated here epigenetic chromatin modification in the stingless bee Melipona scutellaris. We used an ELISA-based methodology to quantify global methylation status and western blot assays to reveal histone modifications. The results evidenced DNA methylation/demethylation events in larvae and pupae, and significant differences in histone methylation and phosphorylation between newly emerged adult queens and workers. The epigenetic dynamics seen in this stingless bee species represent a new facet in the caste determination process in Melipona bees and suggest a possible mechanism that is likely to link a genotype component to the larval diet and adult social behavior of these bees.
RESUMEN
Fipronil (FP) is an insecticide that belongs to the phenylpyrazole chemical family and is used to control pests by blocking GABA receptor at the entrance channel of the chlorine neurons. The aim of this study was to evaluate the mutagenic, recombinogenic and carcinogenic potential of FP. The mutagenic and recombinogenic effects were evaluated using the somatic mutation and recombination test (SMART) on wing cells of Drosophila melanogaster. Third instar larvae from standard (ST) and high bioactivation (HB) crosses were treated with different concentrations of FP (0.3, 0.7, 1.5 or 3.0 × 10-5 mM). The results showed mutagenic effects at all concentrations tested in the HB cross; and all concentrations tested in the ST cross, except at concentration of 0.7 × 10-5 mM. The carcinogenic effect of FP was assayed through the test for detection of epithelial tumor (warts) in D. melanogaster. Third instar larvae from wts/TM3 virgin females mated to mwh/mwh males were treated with different concentrations of FP (0.3, 0.7, 1.5 or 3.0 × 10-5 mM). All these concentrations induced a statistically significant increase in tumor frequency. In conclusion, FP proved to be mutagenic, recombinogenic and carcinogenic in somatic cells of D. melanogaster.