Food consumption, utilization, and detoxification enzyme activity of the rice leaffolder larvae after treatment with Dysoxylum triterpenes

The toxicity and physiological (enzyme and nutritional indices) effect of Dysoxylum triterpenes 3β,24,25-trihydroxycycloartane and beddomei lactone were evaluated on the rice leaffolder Cnaphalocrocis medinalis (Guenée). The LC₅₀ [6.66 ppm (SD = 0.31), 5.79 ppm (SD = 0.33) for 3β,24,25-DHCL and BL, respectively] and LC ₉₀ [14.63 ppm (SD = 0.36), 13.49 ppm (SD = 0.27) for 3β,24,25-DHCL and BL, respectively] were identified by probit analysis. Fourth instars were exposed to various concentrations (1.5, 3, 6, and 12 ppm) of Dysoxylum triterpenes. Results showed that treated larvae exhibited reduced food consumption and enzyme activity. Food consumption, digestion, relative consumption rate, efficiency of conversion of ingested food, efficiency of conversion of digested food, and relative growth rate values declined significantly but the approximate digestibility of treated larvae was significantly higher as a result of treatment (in particular 6 and 12 ppm). Likewise, the gut enzymes acid phosphatases, alkaline phosphatases, and adenosine triphosphatases were significantly inhibited by the Dysoxylum triterpenes. The high biological activity of these triterpenes from Dysoxylum sp. could be used as an active principle during the preparation of botanical insecticides for insect pest like rice leaffolder.     

The toxicity and physiological effect of neem limonoids on Cnaphalocrocis medinalis (Guenée) the rice leaffolder

The effect of neem (Azadirachta indica) limonoids azadirachtin, salannin, deacetylgedunin, gedunin, 17-hydroxyazadiradione, and deaceytlnimbin on gut enzyme activity of the rice leaffolder larvae was investigated. When fed a diet of rice leaves treated with limonoids in bioassays, gut tissue enzymes—acid phosphatases (ACP), alkaline phosphatases (ALP), and adenosine triphosphatases (ATPase) activities of rice leaffolder (Cnaphalocrocis medinalis) larvae are affected. Azadirachtin was most potent in all experiments. Larvae that were chronically exposed to limonoids showed a reduction in weight (59-89%) and exhibited a significant reduction in ACP, ALP, and ATPase activities. These results indicate neem limonoids affects gut enzyme activities. These effects are most pronounced in early instars.     

Effect of biopesticides applied separately or together on nutritional indices of the rice leaffolderCnaphalocrocis medinalis

Laboratory assays were done to evaluate the effect of neem seed kernel extract (Azadirachta indica A. Juss),Vitex negundo L. (Lamiales: Verbenaceae) leaf extract, andBacillus thuringiensis (Berliner), applied separately or together, on nutritional indices of the rice leaffolderCnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae). All three biopesticides suppressed feeding and larval growth and low concentrations affected the larval performance. The combined effect of the three biopesticides resulted in a considerable decrease in nutritional indices, indicating strong deterrence. Dose response relationships were established with respect to frass production and larval growth. The efficiency of conversion of ingested and digested food was considerably reduced.     

Physiological effect of chitinase purified from Bacillus subtilis against the tobacco cutworm Spodoptera litura Fab.

An extracellular chitinase was purified from Bacillus subtilis. The lethal concentration (LC₅₀) was determined by using chitinase in first, second, and third instars of Spodoptera litura Fab. Chitinase showed the highest insecticidal activity at 6 μM concentration within 48 h. The nutritional indices were also significantly affected by the 6 μM concentration (P < 0.05). Food consumption, efficiency of conversion of ingested and digested food, relative growth rate, and consumption values declined significantly while approximate digestibility was increased. Our study indicates that treatment of host plant leaves with the chitinase can regulate (reduce) larval growth and weight, and enhance the mortality. This may serve as an effective biocide and alternative to Bt toxin.     

芳香植物气味及提取液对茶尺蠖行为的影响

为了明确芳香植物对茶尺蠖Ectropisobliqua的驱避、拒食作用和获得应用于茶园茶尺蠖"推-拉"策略的植物材料,利用"Y"型嗅觉仪室内测定其成虫对芳香植物气味的趋性反应,并采用叶碟法测定芳香植物提取液对其幼虫的拒食作用及营养干扰效果。丁香罗勒、迷迭香、柠檬桉和芸香植株挥发物及甲醇提取液对茶尺蠖雌、雄成虫有显著的驱避效果。猫薄荷、迷迭香和鼠尾草的2 00 mg/mL甲醇提取液对茶尺蠖3龄幼虫表现出显著拒食活性,24 h后的选择性拒食率分别为93.85%、86.00%和77.32%,非选择性拒食率分别为85.77%、87.00%和88.05%。其中,迷迭香处理的幼虫生长率最低,仅为0.12 mg/天,对幼虫的相对取食量、食物利用率和食物转化率等营养指标有影响。研究表明,10种芳香植物中迷迭香具有显著的行为调控功能,是构建茶尺蠖"推-拉"策略的理想材料。     

The effects of azadirachtin and nucleopolyhedrovirus on midgut enzymatic profile of Spodoptera litura Fab. (Lepidoptera: Noctuidae)

The effects of azadirachtin (AZA) and nucleopolyhedrovirus (NPV) on midgut enzyme activity in Spodoptera litura Fabricius (Lepidoptera: Noctuidae) (Tobacco cutworm) were evaluated. Gut enzyme activities were decreased by AZA and NPV individually and in combination. When S. litura larvae were fed a diet of castor leaves treated with AZA and NPV in bioassays, gut enzyme—acid phosphatases, alkaline phosphatases, adenosine triphosphatases, and lactate dehydrogenase—activities were decreased. There were statistically significant differences (P ? 0.05) in enzyme activities between combined and individual treatment. A synergistic effect of botanical insecticides and virus was found when combined in low doses. These effects are most pronounced in early instars. Maximum weight loss (59-72%) occurred, when AZA and NPV were combined.     

亚致死剂量甲氨基阿维菌素苯甲酸盐对家蚕食物利用的影响

采用质量分析法研究了经甲氨基阿维菌素苯甲酸盐3个亚致死剂量(LC5、LC10和LC20)药剂处理的桑叶对家蚕Bombyx mori 3、4龄幼虫生长及食物利用的影响。结果表明,亚致死剂量甲氨基阿维菌素苯甲酸盐对家蚕3、4龄幼虫的生长和食物利用有明显的抑制作用。处理后家蚕的生长指标(虫体增重率和相对生长率)、营养指标(相对取食量和排粪量)和营养效应指数(食物利用率和食物转化率)均显著降低,而近似消化率(营养指标)显著升高。其中3、4龄幼虫在处理后第2 d,其相对生长率、相对取食量、食物利用率和食物转化率分别比对照下降了19.6% ~66.1%、13.9% ~23.6%、6.6% ~57.0%和9.3% ~66.2%。     

Effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase in insects

The effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase was studied in Spodoptera frugiperda. Larvae were fed an artificial diet containing fruit extract and their midgut was used for enzyme determination. As compared to the control, consumption of the extract containing diet resulted in a 31% inhibition of the cholinesterase activity and a 34% activation of the NADPH-cytochrome c reductase activity. In the cockroach Leucophaea maderae, the effect on reductase activity was even more pronounced (43%).     

Effect of botanical insecticides and bacterial toxins on the gut enzyme of the rice leaffolderCnaphalocrocis medinalis

The effect of botanical insecticides and bacterial toxins on gut enzyme activity of larvae of the rice leaffolderCnaphalocrocis medinalis (Guenée) (Insecta: Lepidoptera: Pyralidae) was investigated. Gut enzyme activities were affected by botanical insecticides and bacterial toxin individually and in combination. When fed a diet of rice leaves treated with botanical insecticides and bacterial toxins, in bioassays the activities of gut tissue enzymes — acid phosphatases (ACP), alkaline phosphatases (ALP) and adenosine triphosphatases (ATPase) — of rice leaffolder larvae were affected. When combined, the effect was more severe at a low concentration. Larvae that were chronically exposed to botanical insecticides and bacterial toxins showed a reduction in weight (59–89%) and exhibited a significant reduction in ACP, ALP and ATPase activities. The combination ofBacillus thuringiensis kurstaki and botanical insecticides caused a decrease of twofold in enzyme activity even at reduced concentration. A synergistic effect was found when botanical insecticides and bacterial toxins were combined at low doses. These effects were most pronounced in early instars. Clear dose-response relationships were established with respect to enzyme activity. In conclusion: (i) biopesticides are relatively safe and biodegradable; (ii) a synergistic effect of botanical insecticides and bacterial toxins was found; (iii) less expensive, readily available and naturally occurring biopesticides could be an alternative for organic and inorganic pesticides in controlling RLF. http://www.phytoparasitica.org posting Sept. 28, 2004.     

Effect of fraxinellone on growth and digestive physiology of Asian corn borer, Ostrinia furnacalis Guenee

A limonoid, fraxinellone significantly reduced the relative growth rate, food consumption rate as well as the efficiency of conversion of ingested food of the Asian corn borer, Ostrinia furnacalis when incorporated into artificial diets at concentration of 10 ppm and above. After being fed with diets that contained fraxinellone for 24 h, the larval midguts of O. furnacalis possess significantly lower activities of α-amylase and non-specific proteases and higher activities of cytochrome P450s. In vitro, the compound did not inhibit the activities of α-amylase and non-specific proteases extracted from the larval midguts. The modes of action of fraxinellone against the insect were discussed.     

A comparative study of rac- and S-metolachlor on some activities and metabolism of silkworm, Bombyx mori L.

The racemic product of metolachlor contains two R-enantiomers and two S-enantiomers. S-Metolachlor is a more effective herbicidal agent than rac-metolachlor; currently, it is widely used instead of rac-metolachlor. The comparative toxicity of rac- and S-metolachlor was assessed on the basis of their effects on some enzyme activities of the fifth instar of the silkworm, Bombyx mori L. These results suggest that rac-metolachlor and S-metolachlor have different effects on enzyme activities of fifth-instar silkworm larvae. Hemolymph lactate dehydrogenase and catalase activity levels in silkworms treated with rac-metolachlor were much lower than those in silkworms treated with S-metolachlor. Control hemolymph ACP activity was 44-73% higher than that of rac-metolachlor-treated hemolymph, but the difference was not large between S-metolachlor treatment and the control. Midgut alkaline phosphatase activity of silkworms treated with rac-metolachlor was 46% lower than control activity, and the effect of S-metolachlor on midgut alkaline phosphatase activity was minor. The difference between the two herbicides was apparent mainly during the first 5 days of the fifth instar, and then gradually disappeared. It is shown that compared with S-metolachlor, rac-metolachlor is more toxic to economically important silkworms because of its effects on some enzymes and on metabolism.     

Effects of Artemisia annua L. (Asteracea) on nutritional physiology and enzyme activities of elm leaf beetle, Xanthogaleruca luteola Mull. (Coleoptera: Chrysomellidae)

Methanolic extract of Artemisia annua L., a weed collected around paddy fields in north of Iran, was investigated for its toxic effects on: feeding, growth, fecundity, fertility including the biochemical characteristics of elm leaf beetle Xanthogaleruca luteola Mull. The toxicity tests indicate that insecticidal effect was 2.4-fold higher in 3rd instar larvae than in adults. The LC₅₀ was 48% and 43.77% of a methanolic leaf extract at 24 and 48 h, respectively. The feeding deterrence was highest at 10% concentration and lowest at 0.625% for 3rd instar larvae and adults during the first 24 h and decreased thereafter. Larval duration significantly increased at higher concentrations. Larval treatment with 10% methanolic extract resulted in deformed adults (adultoids) with drastic changes in gross morphology of female reproductive system. Food digestion and assimilation significantly decreased in adults. Larval duration and adult fecundity and fertility were also significantly reduced. Twenty-four hours after treating 3rd instar larva with the extract the levels of glucose, protein, urea, uric acid, α-amylase, alkaline phosphatase, alanine amino transferase and aspartate amino transferase significantly changed. However, at 48 h the extract lost its potency. The extract did not effect the level of cholesterol in treated larva. These results indicate that Artemisia annua L. extract contains inhibitors of key metabolic pathways that may be useful in future control of the elm leaf beetle.     

Effects of acute and multiple dose administration of Sumithion (O,O-dimethyl O-(3-methyl-4-nitrophenyl) phosphorothionate) on some enzymes of Periplaneta americana (Linn.)

Acute exposure to Sumithion (65.0 μg/g dose) causes significant inhibition of acetylcholinesterase (AChE) in the digestive tissue of adult P. americana. At this dose there is practically no effect on the activities of either acid or alkaline phosphatases or non-specific esterases. Exposures to multiple doses of 1.3 μg/g at intervals of 24 and 48 hr, respectively, lower AChE activity by approximately 40%. On the other hand multiple doses cause an initial rise in the activities of phosphatases, indicating their possible involvement in the metabolism of the pesticide. However, later on, there is a decrease in levels of acid and alkaline phosphatase activities. This is coupled with a steady rise in nonspecific esterase activity indicating a possible take over of metabolizing role by these enzymes.     

Biological and biochemical responses of Biomphalaria alexandrina to some extracts of the plants Solanum siniacum and Artemisia judaica L.

The molluscicidal activity of cold water, boiled water, methanol, ethanol, acetone and chloroform extracts of Solanum siniacum and Artemisia judaica L. plants against Biomphalaria alexandrina snails was carried out. The tests revealed plant’s ethanol extract was more toxic to the snails than the other tested extracts. Therefore, it was tested against snails’ fecundity (M_x), reproduction rate (R₀) and their infection with Schistosoma mansoni miracidia. In addition, biochemical parameters and the activities of some enzymes in tissues of snails treated with the two tested plants were determined. As well, glucose concentration in snails’ hemolymph was evaluated. Exposure of B. alexandrina snails to plant’s ethanol extract led to a significant reduction in their survival and snails’ fecundity, reproduction rate. In addition, it caused a considerable reduction in the infectivity of S. mansoni miracidia to the snails. Also, it caused a reduction in number of cercariae per snail during the patent period and in the period of cercarial shedding. The results revealed that the glucose concentration in hemolymph and Lactate level in soft tissues of treated snails were increased (P < 0.001) while glycogen, total protein, the lipid content and the pyruvate level in snail’s tissues decreased (P < 0.001). The activities of lactic dehydrogenase (LDH), pyruvatekinase (PK) and cytochrome oxidase (CY) enzymes in homogenate of snail’s tissues were reduced (P < 0.001) in response to treatment with the two tested plants while protease (PR) activity increased (P < 0.001). It is concluded that the application of LC₂₅ of ethanol extract of S. siniacum and A. judaica L. may be helpful in snail control as it interferes with the snails’ biology and physiology.     

Insecticidal action of phytohemagglutinin (PHA) against the grain aphid, Sitobion avenae

The effects of phytohemagglutinin (PHA) on development, fecundity, and mortality of the grain aphid, Sitobion avenae (Fabricius), were determined. Also determined were the effects of PHA on the activity of gut enzymes involved in sugar and phosphorus metabolism (α- and β-glucosidase and alkaline and acid phosphatase) and on the feeding behavior of adult apterae.When added to a liquid diet, PHA increased the pre-reproductive period, decreased fecundity, and increased mortality of S. avenae. PHA also increased the developmental time and reduced the intrinsic rate of natural increase. The aphids fed on sucrose-agarose gels with higher concentrations of PHA lectin exhibited no phloem-feeding behavior. The number of the gel penetrations was reduced and duration of the pathways was increased by higher concentrations of PHA. In addition, activities of α-glucosidase and alkaline phosphatase in homogenates of adult aphids were reduced. The results indicate that PHA has potential in a biotechnological approach to insect management.     

Influence of Atrazine and Roundup pesticides on biochemical and molecular aspects of Biomphalaria alexandrina snails

The excessive use of pesticides in agriculture has sparkled the interest of scientists in investigating the harmful effects of these compounds. The present study evaluates the pesticides Atrazine and Roundup (glyphosate) on biochemical and molecular aspects of Biomphalaria alexandrina snails. The results showed that LC₁₀ of these two pesticides caused considerable reduction in survival rates and egg production of treated snails. Additionally, Atrazine proved to be more toxic to B. alexandrina snails than Roundup. In treated snails, glucose concentration (GL) in the hemolymph as well as lactate (LT) and free amino acid (FAA) in soft tissues of treated snails increased while glycogen (GN), pyruvate (PV), total protein (TP), nucleic acids (DNA and RNA) levels in snail’s tissues decreased. The activities of glycogen phosphorylase (GP), superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR), succinic dehydrogenase (SDH), acetylcholinesterase (AChE), lactic dehydrogenase (LDH) and phosphatases (ACP and ALP) enzymes in homogenate of snail’s tissues were reduced in response to the treatment with the two pesticides while lipid peroxide (LP) and transaminases (GOT and GPT) activity increased (P < 0.001). The changes in the number, position and intensity of DNA bands induced by pesticides may be attributed to the fact that pesticide can induce genotoxicity through DNA damage. It was concluded that the pollution of the aquatic environment by Atrazine and Roundup pesticides, would adversely affect the metabolism of the B. alexandrina snails, and have adverse effects on its reproduction.     

Biochemical alteration induced by monocrotophos in the blood plasma of fish, Channa punctatus (Bloch)

Monocrotophos (MCP), commonly known as azodrin, is one of the organophosphate (OP) pesticides extensively used in agricultural practices throughout the world. Channa punctatus were exposed to sublethal concentrations (0.96 and 1.86 mg/L) of monocrotophos for 15 and 60 days to assess the alterations in the level of some biochemical parameters in blood plasma. Significant alterations in all the biochemical parameters were found to be dose dependent. Hypoglycemia and hypocholesteremia were observed in plasma of fish at both exposure periods (15 and 60 days). Increased activities of glutamate-oxalacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), acid and alkaline phosphatase of blood plasma indicated hepatic tissue damage. Decrease in lactate dehydrogenase (LDH) content in plasma further indicated lower metabolic rate after 60 days of exposure. Significant decline in triglycerides content was observed in fish exposed to both sublethal concentrations of monocrotophos. We suggest that analysis of biochemical parameters in the fish blood may be useful in environmental biomonitoring.     

Effects of Bacillus subtilis metabolites on larval Aedes aegypti L

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 3.71 μg/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.     

Boric acid toxicity to the German cockroach, Blattella germanica: Alterations in midgut structure, and acetylcholinesterase and glutathione S-transferase activity

Oral toxicity of boric acid, an inorganic insecticide, was evaluated on German cockroach, Blattella germanica L. (Dictyoptera, Blattellidae). Newly emerged adults were exposed to various concentrations of boric acid incorporated into the diet. Results showed that treated insects exhibited toxic symptoms with a dose-dependent mortality. Histological study of midgut revealed alterations in the epithelial cells and a significant increase in the epithelium thickness. In a second series of experiments, the compound was investigated on the activities of acetylcholinesterase (AChE) and glutathione S-transferases (GSTs). Data showed that the compound induced GSTs and reduced the activity of AChE. From this experiment, it may be concluded that ingested boric acid caused death of insects perhaps ultimately by starvation via alterations of the midgut. Additionally, the compound seemed to present a neurotoxic action as evidenced by the symptoms of poisoning and the reduction in AchE activity.     

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under in vitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼45% and 30%, respectively. Lipid peroxidation, H₂O₂ content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼64% and 67% in larvae fed on diets with 100 μM of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H₂O₂, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects.