Long-term storage does not affect the infectivity of entomopathogenic nematodes on insect hosts.

Entomopathogenic nematodes (EPNs) are effective biopesticides used in insect control. There are several formulations of entomopathogenic nematodes which are commercially available, but their popularity is low because the requirement of periodic release of stored entomopathogenic nematodes in field has reduced infectivity and is a constraint in large-scale repeated application of entomopathogenic nematodes in field. So, in order to overcome this impediment, the current study focuses on storage of entomopathogenic nematodes for a longer period of time using encapsulating beads. In this study, three EPNs, namely, Steinernema carpocapsae, Steinernema monticolum, and Rhabditis blumi, were encapsulated with Arabic gum and sodium alginate gel beads at different concentrations. Bio-efficacy of stored EPNs at different concentrations (50, 100, 200, 500, and 1000 infective juveniles (IJs)) in varying time intervals of 3, 6, 9, and 12 weeks were tested against Culex quinquefasciatus larvae. The results show that, after 3 weeks of storage, these entomopathogenic nematode species showed survival up to 96%, and in 12-week storage, survival rate was 50%. The virulence of the progeny that emerged from these stored entomopathogenic nematodes was similar to the fresh infective juveniles. The results of the present study show that Arabic gum-sodium alginate combination is a promising approach toward storing entomopathogenic nematodes. Furthermore, semi-field trials showed 70-90% larval mortality at a concentration of 3000 IJs (encapsulated and stored entomopathogenic nematodes in S. monticolum, S. carpocapsae, and R. blumi) on C. quinquefasciatus larvae. KEY POINTS: • Increased survival of stored EPNs (S. carpocapsae, S. monticolum, and R. blumi) in encapsulated gel using Arabic gum, sodium alginate, and yeast cells was assessed. • The efficacy was greater in stored EPNs at different concentrations from 3 to 12 weeks. • Semi-field trials of stored encapsulated EPNs are efficient in mosquito larval control.

Biosynthesis and characterization of silver nanoparticles from symbiotic bacteria Xenorhabdus nematophila and testing its insecticidal efficacy on Spodoptera litura larvae.

Spodoptera litura, one of the polyphagous pests, causes huge economical lose and use of chemical pesticide causes impact to the environmental. The present study deals with the use of cell- free supernatant of bacteria Xenorhabdus nematophila NP-1 strain for synthesizing silver nanoparticles and analyzing its larvicidal ability against Spodoptera litura. Color change from yellow to dark brown specifies the synthesis of AgNPs. UV-Vis spec indicates the presences of AgNPs at 440 nm λmax and functional groups; alcohols, carboxylic acids, aromatics, alkylhalides, ethers and phenols were confirmed by FTIR. SEM revealed the synthesized AgNPs is in spherical shape, EDaX confirms the elemental composition and the crystalline nature were observed using XRD. GC-MS analysis showed presence of Benzencepropanoic acid, 1, 3, 5 Trichloropent-2-ene, 1,1-Dichloro-2,3- dicmethycycloprone and 1,2-benzenedicarboxylic acid bioactive compounds some of which may be responsible for insecticidal and antibacterial activity. The antibacterial activity against S. aureus, B. subtilis and K. pneumoniae showed maximum zone of inhibition at 100 µL/mL. Larvicidal activity of S. litura shows highest mortality at 48 h. In potted plant experiment, AgNPs treated plants showed less damage, with less leaf consumption by S. litura larvae. Further, the synthesis of AgNPs were targeted to zebrafish embryos (non- target organism) and it didn't exhibit any toxic effect even at higher concentration. Our experiment concludes that, AgNPs synthesized using NP-1 strain has highest antimicrobial and insecticidal activity, which can be used in biomedical and biopesticides.

Biogenic synthesis of ZnO nanoparticles mediated from Borassus flabellifer (Linn): antioxidant, antimicrobial activity against clinical pathogens, and photocatalytic degradation activity with molecular modeling.

Borassus flabellifer leaf extract has been used for rapid biogenic synthesis of zinc oxide nanoparticles (ZnO-NPs) due to rich source of bioactive compounds. The synthesized ZnO-NPs were preliminarily confirmed by UV-visible spectroscopy adsorption peak range at 365 nm. The XRD (X-ray diffraction) confirms purity of ZnO-NPs that were crystalline in nature. The analysis of FT-IR (Fourier-transform infrared spectroscopy) confirms the presence of the following functional group such as alcohol, phenols, carboxylic acids, primary amides, secondary amides, and alkyl halide. The Field Emission Scanning Electron Microscope (FE-SEM) analysis indicated that ZnO-NPs were in spherical shape, followed by EDX analysis which confirmed the presence of Zn-element. Antimicrobial effect of ZnO-NPs was investigated using different clinical pathogens like bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella Pneumonia, and Pseudomonas aeruginosa and fungi Aspergillus flavus, Candida albicans, and Penicillium expansum which confirmed ZnO-NPs efficiency as an antimicrobial agent. ZnO-NP antimicrobial efficiency was observed in higher zone of inhibition at 50 µg/mL concentrations. Antioxidant activity was ascertained to be used for several biomedical applications. The ZnO-NPs efficiently degraded the environmental toxic dyes (methylene blue and crystal violet) under sunlight, and up to 95% higher degradation was achieved in both dyes. In support of photo light degradation, the study was progressed to understand the ZnO-dye interaction stability using molecular mechanism, and it shows efficient bonding features in the NPs environment. Overall, this investigation has great potential for being an effective and eco-friendly material used in environmental applications.

Mycosynthesis of bimetallic zinc oxide and titanium dioxide nanoparticles for control of Spodoptera frugiperda.

Spodoptera frugiperda is an important pest of several crops. Use of chemical insecticides in insect control has not been successful. In the present study, bio-nano formulation of bimetallic nanoparticles were synthesized using fungal metabolites from Metarhizium anisopliae and tested for insecticidal activity. Metarhizium anisopliae metabolites was synthesized along with bimetallic nanoparticles at different volumes for bioassay studies. Bimetallic nanoparticles showed colour change from light greenish to white. Synthesis of bimetallic NPs was confirmed by UV-vis spectrophotometer, with absorption peak at 387 nm. Functional groups in the synthesized BMNPs were analyzed by FTIR, which showed the presence of functional amines, carboxylic acids, alkenes, alkyl halides, phosphine oxides, anhydrides, phosphines, sulfonates, acid chlorides and alkynes. XRD analysis confirms the crystalline nature of nanoparticles. SEM analysis exhibits, the nanoparticles to be spherical in shape with size ranging 9.50 nm. EDaX analysis confirmed the elemental composition of synthesized NPs, while DLS for size distribution ranging at 80 to 137 nm. The larvicidal activity of monometallic and mycogenic bimetallic nanoparticles were tested on third instar S. frugiperda larvae. The highest mortality was observed in BMNPs at a concentration of 100 µg/ml after 48 h exposure. Pupicidal and antifeedant activity was also observed in the tested BMNPs. Detoxification enzymes studies of BMNPs showed considerable lowering these enzyme which may result in toxic manifestation in the insect. These results strongly suggest that, bimetallic nanoparticles synthesized using M. anisopilae can be used for protecting the crops against S. furgiperda.

Diet composition has a differential effect on immune tolerance in insect larvae exposed to Mesorhabditis belari, Enterobacter hormaechei and its metabolites.

In insects, diet plays an important role in growth and development. Insects can vary their diet composition based on their physiological needs. In this study we tested the influence of diet composition involving varying concentrations of macronutrients and zinc on the immune-tolerance following parasite and pathogen exposure in Spodoptera litura larvae. We also tested the insecticidal potential of Mesorhabditis belari, Enterobacter hormaechei and its secondary metabolites on Spodoptera litura larvae. The results shows macronutrient composition does not directly affect the larval tolerance to nematode infection. However, Zinc supplemented diet improved the immune tolerance. While larvae exposed to bacterial infection performed better on carbohydrate rich diet. Secondary metabolites from bacteria produced an immune response in dose dependent mortality. The study shows that the larvae maintained on different diet composition show varied immune tolerance which is based on the type of infection.