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The toxicity of a tea saponin (TS) was investigated on mortality, digestive system, detoxifying and antioxidant enzymes of Ephestia kuehniella Zeller. The larvae were fed on a diet containing diffe...

Saponins cause mortality in insects by reducing food absorption and movement in the gut, which may be used to exploit the control of insect pests. In the current study, tea seed saponin (TSS) was extracted and then added to the artificial diets of Helicoverpa armigera. Pre-ovipositional period of the TSS-treated individuals increased while longevity and fecundity decreased compared to control. There was a significant reduction of the treated individuals in the life table parameters of TSS-treated Individuals including net reproduction rate (R

A combination of bioassay and biochemical approaches were used to determine toxicity of Artemisia annua essential oil (AaEO) Pseudococcus viburni. AaEO via leaf dipping bioassay showed LC50 values ...

Toxic effects of a saponin (TSS) from Camellia sinensis L. were investigated on mortality, nutrition, digestion and ecdysteroid receptors of Helicoverpa armigera Hubner through bioassay, biochemical assays and gene expression. Different concentrations of TSS were added to the artificial diet of the larvae to evaluate intended parameters. The LC50 of TSS was found 3.66% with a confidence limit of 3.07–4.51 at 95% although ingestion of sublethal concentration decreased nutritional indices including approximate digestibility, the efficiency of conversion of digested food, the efficiency of conversion of ingested food, relative growth rate, and relative consumption rate. These findings accompanied by the lower activities of digestive enzymes including carbohydrases, TAG-lipase, serine proteases and exopeptidases. Expression of the genes related to digestive enzymes demonstrated a significant decrease in the TSS-ingested larvae similar to biochemical experiment. Moreover, the gene expression of the ecdysteroid receptor in the TSS-treated larvae showed a significant difference compared to control. These results indicate the initial results on the toxicity of TSS against H. armigera and emboss its potential as a nature-origin insecticide.

Efficient control of Chilo suppressalis Walker is always controversial due to highly economic damage, resistance to insecticides and environmental pollutions. So, combination of safe pest controls e.g. biocontrol agents and insect growth regulators seems to be promising via integrated pest management program. Bioassay of hexaflumuron on 4th larval instars revealed concentrations of 44.34, 179.74 and 474.94µg/ml as LC10-50 values. Numbers of total hemocytes, plasmatocytes and granulocytes as well as phenoloxidase activity increased in the different time intervals following treatment by hexaflumuron. Combined effects of hexaflumuron and Beauveria bassiana Vuillemin also increased hemocyte numbers and phenoloxidase activity at different time intervals using all concentrations. Activities of general esterases assayed by α- and β-naphtyl acetate and glutathione S-transferase using CDNB and DCNB increased 1-12h post-treatment. Activities of aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transferase and aldolase increased in the larvae treated by hexaflumuron. However enhanced activity of lactate dehydrogenase was only obtained by treating 180 and 470µg/ml concentrations of hexaflumuron. Activities of ACP and ALP were found to be higher than control for all time intervals even 1-12h post-treatment. The amounts of HDL and LDL increased in the highest concentrations of hexaflumuron after 12-24h of post-treatment. Amount of triglyceride was higher than that of control after 1 and 3h but it was lower in other time intervals. Amounts of glycogen and protein were lower than those of control for all time intervals except for 6 and 12h of post-treatment in case of protein. Results of the current study revealed negative effects of hexaflumuron on intermediary metabolism of Chilo suppressalis but it increased the number of hemocytes and activity of phenoloxidase which are responsible for spore removal from hemolymph. It can be concluded that hexaflumuron is able to decrease survival and biological performance of C. suppressalis via intervening in intermediary metabolism but the given results showed incompatibility of the IGR with possible microbial control.

The effects of pyriproxyfen were determined on the cellular immunity and phenoloxidase activity in the 4th instar larvae of Chilo suppressalis Walker. The bioassay results revealed the effective concentrations of: 10L : 18C, 30L : 72C and 50L : 190C μg · ml−1. The sole effect of 18 and 72 μg · ml−1 concentrations at intervals of 1–3 h caused a higher number of total hemocytes in the treated larvae than the control, but the reverse results were observed after 6–24 h. The number of plasmatocytes was lower than that of the control for intervals of 3–24 h but the number of granulocytes was higher than the control after 1–3 h although no significant differences were observed at the other times. In the treated larvae, the activities of phenoloxidase were higher and lower than those of the control after 1–3 h and 6–24 h, respectively. The combined effects of pyriproxyfen and the entomopathogenic fungus, Beauveria bassiana isolate B3 caused higher numbers of total hemocytes, plasmatocytes, and granulocytes in the treated larvae by use of the three concentrations of pyriproxyfen, at intervals of 6 and 12 h. Although the numbers of nodules in the larvae treated with concentrations of 18 μg · ml−1 were higher than those of other treatments, the overall numbers were lower than those of the control. Finally, the activity of phenoloxidase in the treated larvae was higher than that of the control, at intervals of 6 and 12 h post-treatment. Findings of the current study indicate an intervening role of pyriproxyfen in the cellular immunity of C. suppressalis to entomopathogenic objects.

In the current study, two phenoloxidases (POs) from the larvae of Chilo suppressalis Walker were extracted and purified by column chromatography using Sepharyl G-100 and DEAE-Cellulose fast flow column. Two proteins possessing PO activity, named as POI and POII, were extracted by purification, 5.08- and 5.62-fold, respectively, with 8.94% and 7.31% recoveries, respectively. Also, the specific activities of POI and POII were 0.478 and 0.529 U/mg protein, respectively. Finally, the molecular weights of POI and POII were calculated as 94.6 and 95.7 kDa, respectively. Kinetic parameters of the purified phenoloxidases by Lineweaver-Burk analysis were Vmax of 2.27 and 1.11 U/mg protein and Km of 15.51 and 17.31 mM for POI and POII, respectively. Mg2+ and Cu2+ significantly increased the PO activities. Ca2+ decreased the activity of POI and showed no statistical effects on POII activity. EDTA and DTC significantly inhibited the activities of the purified enzymes, while triethylenetetramine hexaacetic acid (TTHA) and RGTA showed no significant effects on enzymatic activities.

Effects of Pyriproxyfen were determined on intermediary metabolism in the fifth larval instars of Chilo suppressalis via assessments of various enzymatic and non-enzymatic compounds. Bioassay of larvae revealed LC10–50 values of 18, 72 and 190 µg/ml of pyriproxyfen. Results on alanine aminotransferase revealed lower activities in treated larvae versus control for all time intervals while activity of aspartate aminotransferase showed higher activities in treated larvae by 18 and 72 µg/ml of pyriproxyfen. Activities of γ-Glutamyl transferase and aldolase in treated larvae were significantly higher than those of control larvae. Treating of larvae by different concentrations of pyriproxyfen significantly increased lactate dehydrogenase activity for all time intervals although activity of the enzyme after 1 h was not significantly different among control and treated larvae. Activities of acid and alkaline phosphatases in treated larvae were higher than those of control. Amount of high and low density lipoproteins in treated larvae was higher than those of control except for time interval of 3 h of HDL in which control larvae revealed higher amount than 72 and 190 µg/ml treated larvae. Increased activity of the enzyme was noticed in treated larvae versus control in case of general esterases. Similar results were observed in case of glutathione S-transferase. Amount of triacylglyceride, glycogen and protein in control larvae was higher than those of treated larvae for all time intervals indicating their depletion due to energetic costs of pyriproxyfen treatment. These results clearly indicated negative effects of pyriproxyfen on intermediary metabolism of larvae that might lead to desirable mortality in pest population.

Effects of two insect growth regulators (IGRs), hexaflumuron and pyriproxyfen, were studied on the purified phenoloxidase (PO) of Chilo suppressalis. Purification procedure revealed two isozymes of PO, namely POI and POII. IC50 concentrations of hexaflumuron and pyriproxyfen on POI were 0.36, 0.23 μg/ml and on POII were 0.105, 0.42 μg/ml, respectively. Determination of optimal pH and temperature revealed pH 5 and temperature 40 °C as the optimal values for the enzymatic activity. Treating POs with IC50 concentrations of two IGRs was pH and temperature dependent. Effects of these IGRs on POI caused significant increase of Km value versus control suggesting competitive inhibition. Hexaflumuron and pyriproxyfen cause reduction in Vmax value of POII versus control suggesting non-competitive inhibition. The current study shows direct effects of two IGRs on purified PO of C. suppressalis for the first time. These findings could be helpful to develop safe compounds with inhibitory mechanism on PO to neutralise i...