Evaluation of the insecticidal activity of Solanum mammosum (L) fruit extract against Drosophila melanogaster

Solanum mammosum (L.) is known as a poisonous plant and has been extensively used for anti-cancer, antifungal, antibacterial properties. However, the effectiveness of Solanum mammosum against insects has not been fully reported yet. To assess the ability of Solanum mammosum fruit extract against insects, we used fruit fly (Drosophila melanogaster) as a model. In this study, our results showed that phytochemicals and many secondary metabolites were present in Solanum mammosum fruit extracts such as alkaloids, saponins, tannins, terpenoids, coumarins, cardiac glycosides, steroidstriterpenoids, flavonoids, and phenolic compounds. The total polyphenol and flavonoid content were also determined to be 275±1.89 mg GAE/g extract and 676±5.14 mg QE/g extract, respectively. Besides, Solanum mammosum fruit extract caused lethal to 2nd instar, reduced a pupae formation rate of Drosophila larvae. Interestingly, Solanum mammosum extract is more effective in the suppression of pupae eclosion than Ascend as a commercial pesticide. These phenomena might be associated with reduced accumulation of energy, including carbohydrates, lipids, and protein. In addition, the activities of esterases and phosphatases were inhibited in Drosophila by the intake of Solanum mammosum. The study concluded that Solanum mammosum has great potential as an insecticidal agent.

Toxic effects of Drimia maritima (Asparagaceae) ethanolic extracts on the mortality, development, sexual behaviour and oviposition behaviour of Drosophila melanogaster (Diptera: Drosophilidae)

Drimia maritima is a toxic Mediterranean plant and is considered a commercially important drug by its medicinal values and pharmacological properties. In this work, we seek to evaluate the direct and delayed effects of the ethanol extracts of this plant on vinegar fly (Drosophila melanogaster), an excellent laboratory model. The treatment was carried out by ingestion on second-stage larvae (L2) and then we evaluated the impact of this treatment on mortality, development, sexual behaviour, on female's oviposition choice, fecundity, and fertility of adults. The results showed that mortality rates can reach 100% after 15 days of treatment as we recorded an acceleration of the flies' development. After treatment, we observed incomplete nuptial courtship and a significant decrease in eggs laid number and larvae number of the first generation treated. This study indicates that D. maritima ethanolic extract has a neurotoxic property, our results confirmed the presence of toxic secondary metabolites in the extract studied.(AU)

Toxic effects of Drimia maritima (Asparagaceae) ethanolic extracts on the mortality, development, sexual behaviour and oviposition behaviour of Drosophila melanogaster (Diptera: Drosophilidae)

Drimia maritima is a toxic Mediterranean plant and is considered a commercially important drug by its medicinal values and pharmacological properties. In this work, we seek to evaluate the direct and delayed effects of the ethanol extracts of this plant on vinegar fly (Drosophila melanogaster), an excellent laboratory model. The treatment was carried out by ingestion on second-stage larvae (L2) and then we evaluated the impact of this treatment on mortality, development, sexual behaviour, on female's oviposition choice, fecundity, and fertility of adults. The results showed that mortality rates can reach 100% after 15 days of treatment as we recorded an acceleration of the flies' development. After treatment, we observed incomplete nuptial courtship and a significant decrease in eggs laid number and larvae number of the first generation treated. This study indicates that D. maritima ethanolic extract has a neurotoxic property, our results confirmed the presence of toxic secondary metabolites in the extract studied.

Toxic effects of Cleome arabica L. (Capparidaceae) aqueous extracts on mortality and sexual behavior of Drosophila melanogaster (Diptera: Drosophilidae)

Cleome arabica is a spontaneous plant in arid zones,toxic and has hallucinogenic effects. In this work, we seek toevaluate the direct and delayed effect of the aqueous extractson the vinegar fly Dorsophila melanogaster, laboratory modelpar excellence. Treatment is by ingestion on second instarlarvae (L2). The results show that mortality rates can reach90% after 15 days of treatment as we recorded a disturbanceof fly development. Observation of sexual behavior intreatment-emergent adults indicates that C. arabica actsnegatively on the course of the different sequences leading tomating in Drosophila.(AU)

Toxic effects of Cleome arabica L. (Capparidaceae) aqueous extracts on mortality and sexual behavior of Drosophila melanogaster (Diptera: Drosophilidae)

Cleome arabica is a spontaneous plant in arid zones,toxic and has hallucinogenic effects. In this work, we seek toevaluate the direct and delayed effect of the aqueous extractson the vinegar fly Dorsophila melanogaster, laboratory modelpar excellence. Treatment is by ingestion on second instarlarvae (L2). The results show that mortality rates can reach90% after 15 days of treatment as we recorded a disturbanceof fly development. Observation of sexual behavior intreatment-emergent adults indicates that C. arabica actsnegatively on the course of the different sequences leading tomating in Drosophila.

Vernonanthura polyanthes leaves aqueous extract enhances doxorubicin genotoxicity in somatic cells of Drosophila melanogaster and presents no antifungal activity against Candida spp / O extrato aquoso das folhas de Vernonanthura polyanthes potencializa a genotoxicidade da doxorrubicina em células somáticas de Drosophila melanogaster e não apresenta atividade antifúngica contra Candida spp

Vernonanthura polyanthes (Spreng.) A.J. Vega & Dematt. (Asteraceae), known as assa-peixe, has been used in ethnomedicine for the treatment of various diseases such as bronchitis, pneumonia, hemoptysis, persistent cough, internal abscesses, gastric and kidney stone pain. Moreover, some studies demonstrated that species of Genus Vernonia present antifungal activity. Due to the biological relevance of this species, the aim of this study was to investigate the toxic, genotoxic, antigenotoxic and antifungal potential of V. polyanthes leaves aqueous extract in somatic cells of Drosophila melanogaster or against Candida spp. The aqueous extract of the plant showed no toxic, genotoxic and antigenotoxic activity in the experimental conditions tested using the wing somatic mutation and recombination test (SMART/wing). However, when the extract was associated with doxorubicin, used in this work as a positive control, the mutagenic potential of doxorubicin was enhanced, increasing the number of mutations in D. melanogaster somatic cells. In the other hand, no inhibitory activity against Candida spp. was observed for V. polyanthes leaves aqueous extract using agar-well diffusion assay. More studies are necessary to reveal the components present in the V. polyanthes leaves aqueous extract that could contribute to potentiate the doxorubicin genotoxicity.(AU)

Vernonanthura polyanthes (Spreng.) A.J. Vega & Dematt. (Asteraceae), conhecida como assa-peixe, tem sido utilizada na medicina popular para o tratamento de várias doenças, como bronquite, pneumonia, hemoptise, tosse persistente, abcessos internos, afecções gástricas e cálculo renal. Além disso, alguns estudos já demonstraram que espécies do Gênero Vernonia apresentam atividade antifúngica. Devido à relevância biológica dessa espécie, o objetivo deste estudo foi investigar os efeitos citotóxico, genotóxico, antigenotóxico e antifúngico do extrato aquoso das folhas de V. polyanthes em células somáticas de Drosophila melanogaster ou contra Candida spp. O extrato aquoso da planta não apresentou atividade citotóxica, genotóxica e antigenotóxica nas condições experimentais testadas usando o teste de recombinação e mutação somática em asa (SMART-asa). No entanto, quando o extrato foi associado com a doxorrubicina, utilizada neste trabalho como controle positivo, o potencial mutagênico da doxorrubicina foi potencializado, aumentando o número de mutações em células somáticas de D. melanogaster. Por outro lado, nenhuma atividade inibitória contra Candida spp. foi observada utilizando o extrato aquoso das folhas de V. polyanthes por meio do método de difusão em ágar. Mais estudos são necessários para desvendar os componentes presentes no extrato aquoso das folhas de V. polyanthes que possam contribuir para potencializar a genotoxicidade da doxorrubicina.(AU)

Cultivos de células animais visando a altas concentrações celulares: processo em perfusão e suplementação com aminoácidos / Animal cell cultures aiming high cell concentrations: perfusion process and amino acids supplementation

Células animais são alvo de pesquisas visando sua utilização como plataforma para a expressão de proteínas recombinantes, desde vacinas veterinárias até fatores de coagulação para hemofílicos. Exemplos incluem células de inseto Drosophila melanogaster S2 e células de mamífero BHK-21, que vêm sendo estudadas visando a sua utilização para a produção da glicoproteína do vírus da raiva. Independentemente da estratégia de cultivo utilizada, altas concentrações celulares são em geral associadas a uma maior produção da proteína de interesse. O objetivo deste trabalho foi o de investigar estratégias que possibilitariam o cultivo de células animais em altas concentrações celulares. Células de inseto Drosophila melanogaster S2 produtoras da glicoproteína do vírus da raiva foram cultivadas em frascos agitados a 100 rpm e 28, em meio livre de soro SF 900 II suplementado com os aminoácidos asparagina, cisteína, prolina e serina. A adição dos quatro aminoácidos no meio de cultura refletiu em um aumento da concentração celular máxima (XV MÁX) em 16%. Cisteína, quando adicionada isoladamente no meio de cultura, refletiu em uma velocidade específica máxima de crescimento celular (MÁX) 56% maior. Nessa condição, o fator de conversão glicose a célula (YX/GLC) foi 47% maior, indicando um metabolismo de glicose mais eficiente na geração de células. Esses resultados indicam que cisteína é provavelmente substrato limitante do cultivo de células S2AcGPV em meio SF 900 II. [...] Na perfusão, a concentração celular foi cerca de 3 vezes maior do que no cultivo contínuo sem reciclo de células. Provou-se que é possível cultivar células BHK-21 adaptadas a crescimento em suspensão em altas concentrações celulares em escala laboratorial, utilizando biorreator de bancada e spin-filter interno como sistema de retenção celular.

Animal cells have been under research as a platform for the expression of recombinant proteins, ranging from veterinary vaccines to blood coagulation factors for treating hemophilia. Examples include insect Drosophila melanogaster S2 and hamster BHK-21 cells, currently being studied for the production of rabies virus glycoprotein. Regardless of the cultivation strategy, high cell concentrations are usually associated to a higher protein production. Thus, the aim of this research was to investigate animal cell cultivation strategies that would allow higher cell concentrations than those previously reported. Cells of Drosophila melanogaster S2 expressing the rabies virus glycoprotein (S2AcGPV) were cultivated in shake flasks at 100 rpm and 28 , in SF 900 II serum-free medium supplemented with the following amino acids: asparagine, cysteine, proline, and serine. The addition of the four amino acids to the medium increased the maximum cell concentration (XV MAX) in 16%. When only cysteine was added to the medium, the maximum specific growth rate (ÊMAX) was 56% higher. In this condition, the cell yield on glucose (YX/GLC) was 47% higher, indicating a more efficient glucose metabolism. These results show that cysteine is likely a limiting substrate of S2AcGPV cells growing in SF 900 II medium. [...] During perfusion, the cell maintenance coefficient was not negligible and represented 83% of total glucose consumption. These data indicate that the presence of an internal spin-filter may be associated to cell stress. In perfusion, cell concentration was about 3 times higher than that in continuous culture without cell recycle. In conclusion, it was proved that suspension-adapted BHK-21 cells can be cultivated in a laboratory-scale bioreactor with an internal spin-filter, in order to achieve high cell concentrations.

Cultivos de células animais visando a altas concentrações celulares: processo em perfusão e suplementação com aminoácidos / Animal cell cultures aiming high cell concentrations: perfusion process and amino acids supplementation

Células animais são alvo de pesquisas visando sua utilização como plataforma para a expressão de proteínas recombinantes, desde vacinas veterinárias até fatores de coagulação para hemofílicos. Exemplos incluem células de inseto Drosophila melanogaster S2 e células de mamífero BHK-21, que vêm sendo estudadas visando a sua utilização para a produção da glicoproteína do vírus da raiva. Independentemente da estratégia de cultivo utilizada, altas concentrações celulares são em geral associadas a uma maior produção da proteína de interesse. O objetivo deste trabalho foi o de investigar estratégias que possibilitariam o cultivo de células animais em altas concentrações celulares. Células de inseto Drosophila melanogaster S2 produtoras da glicoproteína do vírus da raiva foram cultivadas em frascos agitados a 100 rpm e 28, em meio livre de soro SF 900 II suplementado com os aminoácidos asparagina, cisteína, prolina e serina. A adição dos quatro aminoácidos no meio de cultura refletiu em um aumento da concentração celular máxima (XV MÁX) em 16%. Cisteína, quando adicionada isoladamente no meio de cultura, refletiu em uma velocidade específica máxima de crescimento celular (MÁX) 56% maior. Nessa condição, o fator de conversão glicose a célula (YX/GLC) foi 47% maior, indicando um metabolismo de glicose mais eficiente na geração de células. Esses resultados indicam que cisteína é provavelmente substrato limitante do cultivo de células S2AcGPV em meio SF 900 II. [...] Na perfusão, a concentração celular foi cerca de 3 vezes maior do que no cultivo contínuo sem reciclo de células. Provou-se que é possível cultivar células BHK-21 adaptadas a crescimento em suspensão em altas concentrações celulares em escala laboratorial, utilizando biorreator de bancada e spin-filter interno como sistema de retenção celular. (AU)

Animal cells have been under research as a platform for the expression of recombinant proteins, ranging from veterinary vaccines to blood coagulation factors for treating hemophilia. Examples include insect Drosophila melanogaster S2 and hamster BHK-21 cells, currently being studied for the production of rabies virus glycoprotein. Regardless of the cultivation strategy, high cell concentrations are usually associated to a higher protein production. Thus, the aim of this research was to investigate animal cell cultivation strategies that would allow higher cell concentrations than those previously reported. Cells of Drosophila melanogaster S2 expressing the rabies virus glycoprotein (S2AcGPV) were cultivated in shake flasks at 100 rpm and 28 , in SF 900 II serum-free medium supplemented with the following amino acids: asparagine, cysteine, proline, and serine. The addition of the four amino acids to the medium increased the maximum cell concentration (XV MAX) in 16%. When only cysteine was added to the medium, the maximum specific growth rate (ÊMAX) was 56% higher. In this condition, the cell yield on glucose (YX/GLC) was 47% higher, indicating a more efficient glucose metabolism. These results show that cysteine is likely a limiting substrate of S2AcGPV cells growing in SF 900 II medium. [...] During perfusion, the cell maintenance coefficient was not negligible and represented 83% of total glucose consumption. These data indicate that the presence of an internal spin-filter may be associated to cell stress. In perfusion, cell concentration was about 3 times higher than that in continuous culture without cell recycle. In conclusion, it was proved that suspension-adapted BHK-21 cells can be cultivated in a laboratory-scale bioreactor with an internal spin-filter, in order to achieve high cell concentrations. (AU)