RESUMEN
Widespread use of synthetic pesticides has resulted in the development of insecticide-resistant populations of pests and harmful effects on human health and the environment. There is a need to identify alternative pest management strategies to reduce our reliance on conventional chemical pesticides. In recent years the use of botanical pesticides for protecting crops from insect pests has assumed greater importance. Methanol extract of Alangium salvifolium (L.f.) Wang has potential insecticidal activity against Spodoptera litura Fab. The active fractions were identified through chromatographic techniques as F-IV (Rf value=0.45) and F-VI (Rf value=0.63) and were subjected to GC-MS (GCMATE II). Fifty, 100 and 200ppm of active fractions were applied to fourth instar larvae and the mortality increased with higher concentrations. Relative consumption rate, relative growth rate, efficiency of conversion of ingested food and efficiency of conversion of digested food values all decreased in treated larvae, but approximate digestibility rate increased after treatment. The hydrolytic enzymes, such as acid phosphatase, alkaline phosphatase and the glycolytic enzyme lactate dehydrogenase were inhibited in treated larvae compared with controls. The histopathology study revealed that the epithelial columnar cells were enlarged, completely atrophied; intercellular spaces were swollen, and also noted a cytoplasmic ooze of cell material that mixed with food column. The present study clearly showed the active fractions from A. salvifolium as potential botanicals to control the larvae of S. litura. This is the first report for nutritional indices, enzymatic activities and histological effects of A. salvifolium chemical constituents against S. litura. Thus probably, this will be used as an alternative for synthetic pesticides against the polyphagous pest like S. litura.
Asunto(s)
Alangiaceae , Insecticidas/toxicidad , Extractos Vegetales/toxicidad , Spodoptera/efectos de los fármacos , Fosfatasa Ácida/metabolismo , Fosfatasa Alcalina/metabolismo , Animales , Ingestión de Alimentos/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Insecticidas/química , Intestinos/efectos de los fármacos , Intestinos/enzimología , Intestinos/patología , L-Lactato Deshidrogenasa/metabolismo , Fitoquímicos/análisis , Fitoquímicos/toxicidad , Extractos Vegetales/química , Hojas de la Planta , Spodoptera/anatomía & histología , Spodoptera/crecimiento & desarrollo , Spodoptera/metabolismoRESUMEN
An extracellular chitinase was identified and purified (CS1 and CS2) from Bacillus subtilis. The 16S rRNA sequencing was submitted in GenBank (accession numbers KC336487 and KC412256). The purified crude enzymes were identified through matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The peptide sequences were matched with chitinase sequences. The peak m/z with 1297. 592 and 3094.570 mascot search resulted sequence was blasted with NCBI protein sequences and confirmed that it is a chitinase enzyme. The effects of chitinase on gut enzymes lactate dehydrogenase, acid phosphatase, alkaline phosphatase and adenosine triphosphatase of the tobacco cutworm Spodoptera litura larvae were investigated. At all concentrations tested, chitinase decreased the activities of these gut enzymes relative to the control. When chitinase treated leaves were fed to larvae in bioassays, gut tissue and gut enzymes were affected. The histological study clearly shows the chitinase treated larval gut, peritrophic membrane and epithelial cells were affected significantly. Chitinase isolated from B. subtilis has effectively reduced the gut enzyme activity and growth of S. litura. The chitin based bioformulation may serve as an effective biocide against the polyphagous pest like S. litura.
Asunto(s)
Bacillus subtilis/enzimología , Quitinasas/toxicidad , Tracto Gastrointestinal/efectos de los fármacos , Insecticidas/toxicidad , Spodoptera/efectos de los fármacos , Fosfatasa Ácida/metabolismo , Adenosina Trifosfatasas/metabolismo , Fosfatasa Alcalina/metabolismo , Secuencia de Aminoácidos , Animales , Bacillus subtilis/genética , Secuencia de Bases , Quitinasas/química , Quitinasas/genética , Quitinasas/aislamiento & purificación , Tracto Gastrointestinal/enzimología , Proteínas de Insectos/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Larva/efectos de los fármacos , Larva/enzimología , Larva/crecimiento & desarrollo , Datos de Secuencia Molecular , ARN Bacteriano/análisis , ARN Ribosómico 16S/análisis , Análisis de Secuencia de Proteína , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Spodoptera/enzimología , Spodoptera/crecimiento & desarrolloRESUMEN
The culture supernatant of a strain of Bacillus subtilis isolated from soil samples killed larvae of the mosquito Aedes aegypti. The metabolites produced by B. subtilis were characterized using high performance liquid chromatography (HPLC). Mortality rate was dose-dependent for all larval instars of A. aegypti. Log probit analysis (95% confidence level) revealed an LC50 of 1.73 and an LC90 3.71µg/ml. Molecular weights/masses of B. subtilis metabolites were confirmed using SDS-PAGE analysis. B. subtilis metabolites were confirmed using HPLC analysis. We demonstrate that secondary metabolites from B. subtilis have larvicidal activity against A. aegypti and may be suitable for the control of this and other mosquito vectors of human disease. The larvae to the metabolites, significant reduction in the activities of acetylcholinesterse, α-carboxylesterase, and acid phosphatases were recorded.
Asunto(s)
Aedes/efectos de los fármacos , Bacillus subtilis/metabolismo , Acetilcolinesterasa/metabolismo , Fosfatasa Ácida/metabolismo , Animales , Activación Enzimática/efectos de los fármacos , Proteínas de Insectos/metabolismo , Insecticidas/metabolismo , Insecticidas/farmacologíaRESUMEN
The larvicidal activities of methanolic fractions from Adhatoda vasica leaf extracts were investigated against the bancroftian filariasis vector Culex quinquefasciatus and dengue vector Aedes aegypti. The results indicated that the mortality rates was high at 100, 150, 200 and 250 ppm of methanol extract of fractions III with R (f) value 0.67 and methanol extract of fraction V with R (f) value 0.64 of A. vasica against all the larval instars of C. quinquefasciatus and A. aegypti. The result of log probit analysis (at 95% confidence level) revealed that lethal concentration, LC(50) and LC(90) values were 106.13 and 180.6 ppm for fraction III, 110.6 and 170 ppm for fraction V of C. quinquefasciatus. And, the LC(50) and LC(90) values were 157.5 and 215.5 ppm for fraction III of A. aegypti and 120 and 243.5 ppm for the fraction V of A. aegypti, respectively. All the tested fractions proved to have strong larvicidal activity (doses from 100 to 250 ppm) against C. quinquefasciatus and A. aegypti. In general, second instar was more susceptible than the later instar. The results achieved suggest that, in addition to their ethnopharmacology value, A. vasica may also serve as a natural larvicidal agent.