Systematic induced resistance in Solanum lycopersicum (L.) against vascular wilt pathogen (Fusarium oxysporum f. sp. lycopersici) by Citrullus colocynthis and Trichoderma viride.

The antifungal effects of Citrullus colocynthis extract (Hexane, chloroform, methanol, and water) were tested in vitro on Fusarium oxysporum f. sp. lycopersici (Sacc.) W. C. Snyder & H. N. Hans (FOL), the causal agent of Fusarium wilt. Of these, methanol and water extract at 10% showed the highest inhibition of mycelial growth of FOL by 12.32 and 23.61 mm respectively. The antifungal compounds were identified through Fourier transform infrared (FT-IR) spectroscopy and gas chromatography-mass spectroscopy (GC-MS). The methanol extract was compatible with the biocontrol agent Trichoderma viride. The antagonistic fungi were mass-cultured under laboratory conditions using sorghum seeds. Both T. viride and C. colocynthis methanol extract was also tested alone and together against FOL under both in vitro and in vivo conditions. The combination of T. viride and C. colocynthis showed the highest percentage of antifungal activity (82.92%) against FOL under in vitro conditions. This study revealed that induced systemic resistance (ISR) in enhancing the disease resistance in tomato plants against Fusarium wilt disease. The combined treatment of T. viride and C. colocynthis significantly reduced the disease incidence and index by 21.92 and 27.02% in greenhouse conditions, respectively. Further, the induction of defense enzymes, such as peroxidase (PO), polyphenol oxidase (PPO), ß-1,3-glucanase, and chitinase were studied. The accumulation of defense enzyme was greater in plants treated with a combination of T. viride and C. colocynthis compared to the control. Reduction of wilt disease in tomato plants due to the involvement of defense-related enzymes is presumed through this experiment.

Efficacy of Precocene I from Desmosstachya bipinnata as an Effective Bioactive Molecules against the Spodoptera litura Fab. and Its Impact on Eisenia fetida Savigny.

The sustainability of agroecosystems are maintained with agro-chemicals. However, after more than 80 years of intensive use, many pests and pathogens have developed resistance to the currently used chemistries. Thus, we explored the isolation and bioactivity of a chemical compound, Precocene I, isolated from the perennial grass, Desmosstachya bipinnata (L.) Stapf. Fractions produced from chloroform extractions showed suppressive activity on larvae of Spodoptera litura (Lepidoptera: Noctuidae), the Oriental armyworm. Column chromatography analyses identified Precocene I confirmed using FTIR, HPLC and NMR techniques. The bioactivity of the plant-extracted Dp-Precocene I was compared to a commercially produced Precocene I standard. The percentage of mortality observed in insects fed on plant tissue treated with 60 ppm Db-Precocene I was 97, 87 and 81, respectively, for the second, third and fourth instar larvae. The LC50 value of third instars was 23.2 ppm. The percentages of survival, pupation, fecundity and egg hatch were altered at sub-lethal concentrations of Db-Precocene I (2, 4, 6 and 8 ppm, sprays on castor leaves). The observed effects were negatively correlated with concentration, with a decrease in effects as concentrations increased. Distinct changes in feeding activity and damage to gut tissues were observed upon histological examination of S. litura larvae after the ingestion of Db-Precocene I treatments. Comparative analyses of mortality on a non-target organism, the earthworm, Eisenia fetida, at equal concentrations of Precocene I and two chemical pesticides (cypermethrin and monocrotophos) produced mortality only with the chemical pesticide treatments. These results of Db-Precocene I as a highly active bioactive compound support further research to develop production from the grass D. bipinnata as an affordable resource for Precocene-I-based insecticides.

Toxicity and developmental effect of cucurbitacin E from Citrullus colocynthis L. (Cucurbitales: Cucurbitaceae) against Spodoptera litura Fab. and a non-target earthworm Eisenia fetida Savigny.

Pest management with natural botanical insecticides is a significant implementation for the sustainability of agroecosystem by reducing the unnecessary risk from the inputs of synthetic insecticides. In this research, we isolated the bioactive compound cucurbitacin E from Citrullus colocynthis (L.) Schrad, and their toxicological effects were screened against different larval instars of Spodoptera litura. The bioactive compound cucurbitacin E was chemically characterized through TLC, FT-IR, and NMR analyses. The larval mortality bioassay revealed that the larvae exposed to cucurbitacin E at the discriminating dose of 50 ppm display higher mortality rate against second (93.8%), third (86.4%), and fourth (73.2%) instar respectively. The lethal concentrations (LC50 and LC90) was detected as 15.84 and 67.60 ppm for third instar respectively. The sub-lethal concentration of cucurbitacin E (2, 4, and 6 ppm) intentionally altered the percentage of survival, pupation, fecundity, and egg hatchability of S. litura. Moreover, antifeedant activity of cucurbitacin E was analyzed using choice-based test. In addition, we found the toxic effects of cucurbitacin E (50 and 100 ppm) and chemical pesticides (cypermethrin and monocrotophos) against terrestrial beneficial earthworm Eisenia fetida, and the result revealed that cucurbitacin E has no harmful effect on non-target organism. Hence, the present study reveals that cucurbitacin E might be a part of a new biorational product alternative to synthetic pesticides.

Toxic effect of essential oil and its compounds isolated from Sphaeranthus amaranthoides Burm. f. against dengue mosquito vector Aedes aegypti Linn.

Aedes aegypti is a major mosquito vector that can transfer many deadly diseases such as dengue, chikungunya, Zika, and yellow fever viruses. Due to the developing resistance among the vector populations by the application of chemical insecticides, alternative eco-friendly vector management strategies are being focused. In this aspect, the present study was carried out to evaluate the mosquitocidal potentials of essential oil of Sphaeranthus amaranthoides (EO-Sa). EO-Sa was found to be effective against Ae. aegypti mosquito vector by exhibiting significant larvicidal, adulticidal and repellent activities. GCMS analysis of EO-Sa revealed the presence of Carvone as the major component (peak area of 89.7%). The larvicidal bioassays performed revealed that the second instar larvae were relatively more susceptible (94.32% mortality) to EO-Sa treatments (75 ppm), LC50, 20.38 ppm.The sub lethal treatment concentration (20 ppm) significantly affected the oviposition, fecundity and morphology of Ae. aegypti. At sub lethal treatment concentration, EO-Sa down regulated α- and ß carboxylesterase and up regulated the GST and CYP450 level of third and fourth instar larvae. Thus the present results illustrates that EO-Sa can deliver a durable larvicidal, repellent and adulticidal activity against Ae. aegypti in an effective and eco-friendly manner.

Comparative efficacy of two mycotoxins against Spodoptera litura Fab. And their non-target activity against Eudrilus eugeniae Kinb.

Entomopathogenic fungi are feasible and effective against the agricultural pest. In the current research we investigated the bioactive comparison of two widely accepted entmopathogens (Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae, (basionym)) against the Spodoptera litura (Fab.) through the assessment of larval tolerance and regulation of antioxidants and non-target impact on the earth worm, E. eugeniae, along with commercial pesticides. The entomopathogenic fungus exposure resulted in the modification of the levels of detoxification enzymes as well as significant increases in catalase and superoxide dismutase activity after exposure to the entomopathogenic fungus. Bioassay results showed that B. bassiana and M. anisopliae displayed larval mortality against third and fourth instars. Correspondingly, sub-lethal concentrations of B. bassiana showed development impairment as compared to M. anisopliae. Gut-histology revealed that mycotoxins dosage (4 × 105) showed significant changes in the midgut tissues as compared to control larvae. The non-target screening through artificial soil assay on the earth worm E. eugeniae, with mycotoxins B. bassiana (5 × 108 conidia/ml/kg) and M. anisopliae (5 × 108 conidia/ml/kg) showed less toxicity as compared to Monocrotophos (10 ppm/kg). Current results suggest that the fungal mycotoxins of M. anisopliae and B. bassiana significantly reduce the development of lepidopteran pests, while having only lesser impact on beneficial earthworms.

Aspergillus flavus (Link) toxins reduces the fitness of dengue vector Aedes aegypti (Linn.) and their non-target toxicity against aquatic predator.

Mosquito that accountable for dispersal of dengue fever is Aedes aegypti Linn. and considered to be a chief vector for dengue especially in South Asian countries. Aspergillus flavus is considered to be wild growing green yellow colonies and synthesis highly regulating aflatoxins (B1, B2, G1 and G2) as a secondary metabolite. Mycotoxins of A. flavus showed its efficacy against III and IV instars of Ae. aegypti with more than 90% mortality at the prominent dosage of 2 × 108 conidia/ml. The proximate lethal concentrations (LC50 and LC90) of mycotoxins against third and fourth instars was 2 × 105 and 2 × 107 respectively. Correspondingly, sub-lethal dosage of mycotoxin A. flavus significantly inhibited the level of α- ß-carboxylesterase and SOD activity and upregulated the level of major detoxifying enzymes GST and CYP450. Moreover, sub-lethal dosage also showed higher deterrent and fecundity effects. Gut-histological examination reveals that the A. flavus considerably affected the gut epithelial cells along with the inner gut lumen as compared to the control. The non-target screening of A. flavus against two aquatic predators (A. bouvieri and Tx. splendens) display more than 80% of mortality rate against both the species at the dosage of 2 × 1016 (two-fold-higher dosage used in larval assays). Thus the biosafety assessment suggests that A. flavus display higher toxicity against the non-targets and it is not-recommended to apply it directly to the aquatic habitat of dengue mosquito which shares their living space with other beneficial insects.

Effect of Aspergillus flavus on the mortality and activity of antioxidant enzymes of Spodoptera litura Fab. (Lepidoptera: Noctuidae) larvae.

Insects have developed tolerance against mycoses caused by entomopathogenic fungi through several humoral and cellular mechanisms. Antioxidant enzymes such as superoxide dismutase, lipid peroxidase, and peroxidase can play a role in defense against mycosis, but the physiological interactions between the fungus and the insect are not well characterized. In this study, the effects of infection by entomopathogenic fungus, Aspergillus flavus on the antioxidant defense system of Spodoptera litura, were investigated. The fungi, A. flavus exposure resulted in modification of the levels of antioxidant enzymes, as well as significant decline in phenoloxidase titers and the total hemocyte count 48 h post exposure. A significant increase was observed in detoxifying enzymes. All these results suggest that A. flavus infects S. litura by directly acting on the immune system, resulting in decreased immune function. Bioassay results showed that A. flavus affects third and fourth instar larvae of S. litura. This report supports the importance of A. flavus as a candidate for biological control of S. litura.

Botanical essential oils and uses as mosquitocides and repellents against dengue.

Plants naturally produce bioactive compounds along with many secondary metabolites which serve as defensive chemical against herbivorers including insect pests. One group of these phytochemicals are the 'Essential Oils' (EO's), which possess an extensive range of biological activity especially insecticidal and insect repellents. This review provides a comprehensive viewpoint on potential modes of action of biosafety plant derived Essential Oils (EO's) along with their principal chemical derivatives against larvae and adult mosquito vectors of dengue virus. The development and use of Essential Oils (EO's) effectively applied in small rural communities provides an enormous potential for low cost effective management of insect vectors of human pathogens which cause disease.

Target and non-target response of Swietenia Mahagoni Jacq. chemical constituents against tobacco cutworm Spodoptera litura Fab. and earthworm, Eudrilus eugeniae Kinb.

Toxicological screening of Swietenia mahagoni Jacq. (Meliaceae, West Indies Mahogany) against the lepidopteran pest Spodoptera litura was examined. Phytochemical screening through GC-MS analysis revealed nine peaks with prominent peak area % in Bis (2-ethylhexyl) phthalate (31.5%) was observed. The larvae exposed to discriminating dosage of 100 ppm deliver significant mortality rate compare to other treatment concentrations. The lethal concentrations (LC50 and LC90) was observed at the dosage of 31.04 and 86.82 ppm respectively. Sub-lethal concentrations (30 ppm) showed higher larval and pupal durations. However, pupal weight and mean fecundity rate reduced significantly. Similarly, the adult longevity reduced significantly in dose dependent manner. Midgut histology studies showed that the methanolic extracts significantly disturbs the gut epithelial layer, lumen and brush border membrane compare to the control. The soil assay on a non-target beneficial organism, the soil indicator earthworm Eudrilus eugeniae, with extracts from S. mahagoni (200 mg/kg) showed no toxicity compared to Monocrotophos at the dosage of 10 ppm/kg. Current results suggest that this bio-rational plant product from S. mahagoni displays a significant effect to reduce lepidopteran pests with low toxicity to other beneficial species.

Toxicological effects of chemical constituents from Piper against the environmental burden Aedes aegypti Liston and their impact on non-target toxicity evaluation against biomonitoring aquatic insects.

Dengue is the most rapidly spreading mosquito-borne viral disease in the world. The mosquito, Aedes aegypti, also spreads Yellow fever, Chikungunya, and Zika virus. As the primary vector for dengue, Ae. aegypti now occurs in over 20 countries and is a serious concern with reports of increasing insecticide resistance. Developing new treatments to manage mosquitoes are needed. Formulation of crude volatile oil from Piper betle leaves (Pb-CVO) was evaluated as a potential treatment which showed larvicidal, ovipositional, and repellency effects. Gut-histology and enzyme profiles were analyzed post treatment under in-vitro conditions. The Pb-CVO from leaves of field collected plants was obtained by steam distillation and separated through rotary evaporation. The Pb-CVO were evaluated for chemical constituents through GC-MS analyses revealed 20 vital compounds. The peak area was establish to be superior in Eudesm-7(11)-en-4-ol (14.95%). Pb-CVO were determined and tested as four different concentrations (0.25, 0.5, 1.0, and 1.5 mg/L) of Pb-CVO towards Ae. aegypti. The larvicidal effects exhibited dose dependent mortality being greatest at 1.5 mg Pb-CVO/10 g leaves. The LC50 occurred at 0.63 mg Pb-CVO/L. Larva of Ae. aegypti exposed to Pb-CVO showed significantly reduced digestive enzyme actions of α- and ß-carboxylesterases. In contrast, GST and CYP450 enzyme levels increased significantly as concentration increased. Correspondingly, oviposition deterrence index and egg hatch of Ae. aegypti exposed to sub-lethal doses of Pb-CVO demonstrated a strong effect suitable for population suppression. Repellency at 0.6 mg Pb-CVO applied as oil had a protection time of 15-210 min. Mid-gut histological of Ae. aegypti larvae showed severe damage when treated with 0.6 mg of Pb-CVO treatment compared to the control. Non-toxic effects against aquatic beneficial insects, such as Anisops bouvieri and Toxorhynchites splendens, were observed at the highest concentrations, exposed for 3 h. These results suggest that the Pb-CVO may contain effective constituents suitable for development of new vector control agents against Ae. aegypti.

Development of an eco-friendly mosquitocidal agent from Alangium salvifolium against the dengue vector Aedes aegypti and its biosafety on the aquatic predator.

Plant extracts with their enriched chemical constituents have established potential alternative mosquito control agents. In this research, we developed an eco-friendly mosquitocidal agent from Alangium salvifolium leaves against the dengue and Zika virus vector Aedes aegypti and we investigated its biosafety on the mosquito aquatic predator Toxorhynchites splendens. Results showed that the methanolic extract of A. salvifolium leaves was composed by eight main compounds, with major peak area for hexadecenoic acid (21.74%). LC50 and LC90 values calculated on Ae. aegypti fourth instar larvae were 104.80 and 269.15 ppm respectively. The methanolic extract tested at 100 ppm decreased the α-ß carboxylesterase and SOD ratio significantly and upregulated the GST and CYP450 level. The A. salvifolium methanolic extract displayed significant repellent and adulticidal activity at 100 and 400 ppm respectively. The treatment with 100 ppm of the methanolic extract led to 210 min of protection from Ae. aegypti bites. Four hundred parts per million of the extract showed 98% adult mortality within 30 min from the treatment. Lastly, biosafety assays on the mosquito aquatic predator Tx. splendens showed that the toxicity of the A. salvifolium extract was significantly lower if compared to the cypermethrin-based treatments. The methanolic extract of A. salvifolium showed a maximum of 47.3% mortality rate at the concentration of 1000 ppm, while 0.7 ppm of cypermethrin achieved 91.3% mortality rate on Tx. splendens. Overall, our study enhances basic knowledge on how to improve natural larvicidal agents against dengue and Zika virus mosquito vector with harmless responses on non-target aquatic predators.

Toxicological effects of Sphaeranthus indicus Linn. (Asteraceae) leaf essential oil against human disease vectors, Culex quinquefasciatus Say and Aedes aegypti Linn., and impacts on a beneficial mosquito predator.

Use of environmentally friendly, decomposable natural products for effective vector control has gained considerable momentum in modern society. In this study, essential oil of Sphaeranthus indicus (Si-EO) was extracted and further phytochemical screening revealed fourteen compounds with prominent peak area percentage of 24.9 and 22.54% in 3,5-di-tert-butyl-4-hydroxybenzaldehyde and benzene,2-(1,1-dimethylethyl)-1,4-dimethoxy, respectively. The Si-EO was further evaluated for their larvicidal response against Culex quinquefasciatus and Aedes aegypti at different dosages (62.5, 125, 250 and 500 ppm). The Si-EO displayed prominent larvicidal activity at higher concentration (500 ppm) against both species of mosquitoes. The LC50 and LC90 values of oils were observed at 130 and 350 ppm against C. quinquefasciatus larvae and at 140 and 350 ppm against A. aegypti larvae, respectively. Repellent bioassay established higher protection rate at 200 ppm up to 120 min against both the mosquitoes. However, adulticidal response displayed higher mortality rate only at 700 and 800 ppm against C. quinquefasciatus and A. aegypti, respectively. Toxicological screening against mosquito predator Toxorhynchites splendens revealed that the Si-EO was harmless even at the concentration of 1500 ppm. Overall, these results suggest that the Si-EO plays a significant role as a new bio-rational product against ecological burden mosquito vectors which provides an eco-friendly alternative to synthetic pesticides.

Acute toxicity of chemical pesticides and plant-derived essential oil on the behavior and development of earthworms, Eudrilus eugeniae (Kinberg) and Eisenia fetida (Savigny).

Comparative toxicity of two chemical pesticides (temephos and monocrotophos) versus a plant-derived betel leaf oil Piper betle (L.) to earthworm Eudrilus eugeniae (Kinberg) and redworm Eisenia fetida Savigny, historically: Eisenia foetida (Savigny 1826), was evaluated. Mortality rate was more prominent in temephos at 100 µg concentration to both the earthworms in filter paper test (FPT) as well as 10 mg concentration in artificial soil test (AST). In contrast, P. betle does not display much mortality rate to both the earthworms even at 1000 mg of treatment concentrations. The lethal concentration (LC50) value was observed at 3.89 and 5.26 mg/kg for temephos and monocrotophos against E. eugeniae and 3.81 and 5.25 mg/kg to E. fetida, respectively. Whereas, LC50 value of betel leaf oil was only observed at 3149 and 4081 mg/kg to E. eugeniae and E. fetida, respectively. Correspondingly, the avoidance or attraction assay also displayed that earthworms were more sensitive to the soil containing chemical pesticides. Whereas, the avoidance percentage was decreased in the P. betle oil. Similarly, sublethal concentration of chemical pesticides (5 and 6.5 mg) significantly reduced the earthworm weight and growth rate. However, P. betle oil did not change the developmental rate in the duration of the assay (2, 7 and 14 days) even at 4000 mg treatment concentration. The enzyme ratio of CAT and SOD was also affected significantly after exposure to the chemical pesticides (6.5 mg/kg). Hence, our study implied the risk assessment associated with the chemical pesticides and also recommends plant-derived harmless P. betle oil against beneficial species as an alternative pest control agent.

Chemicals isolated from Justicia adhatoda Linn reduce fitness of the mosquito, Aedes aegypti L.

Extracts from Justicia adhatoda L. (Acanthaceae) strongly reduced the fitness of the mosquito, Aedes aegypti Linn. The methanolic extracts inhibited several enzymes responsible for protecting insects from oxidative and other damage, including glutathione-S-transferase, superoxide dismutase, cytochrome P450, and α- and ß-esterases. They increased repellency (maximum repellency at 100 ppm) in host-seeking adult females using the "arm-in cage assay." Histopathological examination showed the extracts led to serious midgut cell damage. Justicia adhatoda extracts led to reduced fecundity and oviposition of gravid females compared to controls. The extracts led to substantially reduced A. aegypti survival. We infer that the extracts have potential to reduce pathogen transmission by suppressing population growth of A. aegypti, and possibly other mosquito species.

Comparative analysis of mosquito (Diptera: Culicidae: Aedes aegypti Liston) responses to the insecticide Temephos and plant derived essential oil derived from Piper betle L.

Resistance to treatments with Temephos or plant derived oil, Pb-CVO, between a field collected Wild Strain (WS) and a susceptible Laboratory Strain (LS) of Ae. aegypti were measured. The Temephos (0.1mg/L) showed the greatest percentage of mosquito mortality compared to Pb-CVO (1.5mg/L) in LS Ae. aegypti. However, WS Ae. aegypti was not significantly affected by Temephos (0.1mg/L) treatment compare to the Pb-CVO (1.5mg/L). However, both strains (LS and WS) when treated with Pb-CVO (1.5mg/L) displayed steady larval mortality rate across all instars. The LC50 of Temephos was 0.027mg in LS, but increased in WS to 0.081mg/L. The LC50 of Pb-CVO treatment was observed at concentrations of 0.72 and 0.64mg/L for LS and WS strains respectively. The enzyme level of α- and ß-carboxylesterase was reduced significantly in both mosquito strains treated with Pb-CVO. Whereas, there was a prominent deviation in the enzyme ratio observed between LS and WS treated with Temephos. The GST and CYP450 levels were upregulated in the LS, but decreased in WS, after treatment with Temephos. However, treatment with Pb-CVO caused both enzyme levels to increase significantly in both the strains. Visual observations of the midgut revealed cytotoxicity from sub-lethal concentrations of Temephos (0.04mg/L) and Pb-CVO (1.0mg/L) in both strains of Ae. aegypti compared to the control. The damage caused by Temephos was slightly less in WS compared to LS mosquito strains.

Impact of Terminalia chebula Retz. against Aedes aegypti L. and non-target aquatic predatory insects.

Aedes aegypti Linn is one of the most important mosquito species. The vectors are responsible for causing deadly diseases like dengue and dengue hemorrhagic fever. Several chemical pesticides used to control these dengue vectors caused severe toxic significances on human health and other non-target beneficial insects. Therefore the current investigation has been made to access the bio-efficacy of the crude seed extracts of T. chebula against the dengue vector Ae. aegypti. The GC-MS analysis of crude seed extracts of T. chebula identified nine chemical compounds with major peak area in the 1,2,3-Benzenetriol (61.96%), followed by Tridecanoic acid (09.55%). Ae. aegypti larvae showed dose dependent mortality rate was observed between the treatments. Prominent protection rate at greater concentrations of 100ppm and moderate protection at 75 and 50ppm was observed in the repellent assay. Lethal concentration (LC50 and LC90) of fourth instar larvae of Ae. aegypti was observed in 138 and 220ppm concentration respectively. Similarly, the seed extracts showed 100% adulticidal activity at the concentration of 400ppm at 30min of exposure time. Phytochemicals present in the seed extracts of T. chebula significantly affects the major portions of the midgut tissues of Ae. aegypti at the concentration of 100ppm. The toxicological evaluation of seed extracts also proved non-toxic towards the A. bouvieri and Tx. splendens aquatic predatory insects. Hence, the present result suggest that bio-rational plant derived T. chebula could be incorporated in the dengue vector control and have no adverse effects on non-target beneficial insects.

Potential mode of action of a novel plumbagin as a mosquito repellent against the malarial vector Anopheles stephensi, (Culicidae: Diptera).

Plumbagin was isolated and characterized from the roots of Plumbago zeylanica using chromatography: TLC, Column chromatogram, HPLC, FTIR and 1H NMR. The isolated pure compounds were assayed for potency as inhibitors of: acetylcholine esterase (AchE), glutathione S-transferases (GST), superoxide dismutase (SOD), cytochrome P450 and α, ß-esterase, and for repellency with Anopheles stephensi at four different concentrations (25, 50, 75 and 100ppm). The enzyme assay against the pure compound reveals that the level of esterase and SOD was decreased significantly in contrast the level of GST and cytochrome P450 was increased significantly. Our results suggests that novel Plumbagin has significantly alters the level of enzyme comparable to the control. Evaluations resulted in Plumbagin producing maximum repellency scores against An. stephensi mosquitoes in dose dependent manner with highest repellence was observed in the 100ppm. Histological examination showed that the midgut, hindgut and muscles are the most affected tissues. These tissues affected with major changes including separation and collapse of epithelial layer and cellular vacuolization. The results support the utility of plant compound Plumbagin for vector control as an alternative to synthetic insecticides, however, more vigorous field trials are needed to determine viability under natural conditions.

Toxicity and physiological effect of quercetin on generalist herbivore, Spodoptera litura Fab. and a non-target earthworm Eisenia fetida Savigny.

A novel flavonoid, quercetin, was isolated from the medicinal plant Euphorbia hirta L. through chromatography techniques including: TLC, Column chromatography, NMR and then screened for toxicity to larvae of Spodoptera litura Fab. Bioassays were used to analyze pupal weight, survival rate, fecundity, egg hatchability, population growth index, Nutritional index and histopathology of treated larvae at a range of E. hirta extract concentrations. Results of toxicity assays demonstrated that, 6 ppm of quercetin caused 94.6% mortality of second, 91.8% of third, 88% of fourth, and 85.2% of fifth instars respectively. The lethal concentrations (LC50 and LC90) was calculated as 10.88 and 69.91 ppm for fourth instar larvae. The changes in consumption ratio and approximate digestibility produced a reduction in growth rates. Histopathology examinations revealed that the cell organelles were severely infected. Analyses of earthworm toxicity effects resulted in significantly lower rates compared to synthetic insecticides (chloropyrifos and cypermethrin). These results suggests that the botanical compound (quercetin), could have a part as a new biorational product which provides an ecofriendly alternative. Validation of the potential of quercetin, still needs to be demonstrated under field conditions, where formulation will be important in maintaining the activity.

Target and non-target toxicity of botanical insecticide derived from Couroupita guianensis L. flower against generalist herbivore, Spodoptera litura Fab. and an earthworm, Eisenia foetida Savigny.

Botanical insecticides may provide alternatives to synthetic insecticides for controlling Spodoptera litura (F.) and they are target specific, biodegradable, and harmless to mammals. Eight natural chemical compounds with larvicidal activity were identified from fraction F6 of C. guianensis flower extract. Probit analysis of 95% confidence level exposed an LC50 of 223ppm against S. litura third instar larvae. The growth and development of S. litura was affected in sub-lethal concentrations of fraction F6 (50, 100, 150 and 200ppm) compared to controls. Similarly nutritional indices values decreased significantly compared to controls. Fraction F6 also damaged the gut epithelial layer and brush border membrane (BBM). This study also resolved the effects of toxicity to non-target earthworm treated with fraction F6 and chemical pesticides (monotrophos and cypermethrin) and the results showed that fraction F6 had no harmful effect on E. fetida. Further, fraction F6 was eluted and sub fractions F6c (50ppm) showed high mortality against S. litura third instar larvae. Octacosane from fraction F6c was established and confirmed using IR spectrum and HPLC. The time of retention of fraction F6c was confirmed with the octacosane standard. Fraction F6 of C. guianensis extract caused dose-dependent mortality towards S. litura. Octacosane in fraction F6c was establish to be the prominent chemical compound associated with causing mortality but other compounds present in the fraction F6 were shown to be associated with changes in development of S. litura at low dosages. S. litura at low dosage. Therefore, these findings suggest that octacosane may be one of the major insecticidal compounds affecting S. litura survival.

Anti-dengue efficacy of bioactive andrographolide from Andrographis paniculata (Lamiales: Acanthaceae) against the primary dengue vector Aedes aegypti (Diptera: Culicidae).

The current study investigated the toxic effect of the leaf extract compound andrographolide from Andrographis paniculata (Burm.f) against the dengue vector Ae. aegypti. GC-MS analysis revealed that andrographolide was recognized as the major chemical constituent with the prominent peak area compared with other compounds. All isolated toxic compounds were purified and confirmed through RP-HPLC against chemical standards. The larvicidal assays established at 25ppm of bioactive compound against the treated instars of Ae. Aegypti showed prominent mortality compared to other treated concentrations. The percent mortality of larvae was directly proportional to concentration. The lethal concentration (LC50) was observed at 12ppm treatment concentration. The bioactive andrographolide considerably reduced the detoxifying enzyme regulations of α- and ß- carboxylesterases. In contrast, the levels of GST and CYP450 significantly increase in a dose dependent manner. The andrographolide also showed strong oviposition deterrence effects at the sub-lethal dose of 12ppm. Similarly, the mean number of eggs were also significantly reduced in a dose dependent manner. At the concentration of 12ppm the effective percentage of repellency was greater than 90% with a protection time of 15-210min, compared with control. The histopathology study displayed that larvae treated with bioactive andrographolide had cytopathic effects in the midgut epithelium compared with the control. The present study established that bioactive andrographolide served as a potential useful for dengue vector management.