Botanical and synthetic pesticides alter the flower visitation rates of pollinator bees in Neotropical melon fields.

The ecological and economic contributions of pollinator bees to agricultural production have been threatened by the inappropriate and excessive use of pesticides. These pesticides are often applied in areas with ecological peculiarities (e.g., the Neotropical savannah-like region termed as Cerrado) that were not considered during the product development. Here, we conducted field experiments with melon (i.e., Cucumis melo L.) plants cultivated under Brazilian Cerrado conditions and evaluated the impacts of botanical (i.e., neem-based insecticide) and synthetic (i.e., the pyrethroid insecticide deltamethrin and the fungicides thiophanate-methyl and chlorothalonil) pesticides on the flower visitation rates of naturally occurring pollinator bees. Our results revealed that both honey bees (i.e., Apis mellifera L.) and non-Apis bees visited melon flowers and the intensity of bee visitation was moderately correlated with yield parameters (e.g., number of marketable fruits and fruit yield). Pesticide treatments differentially affected bee species. For instance, Plebeia sp. bees were not affected by any pesticide treatment, whereas both A. mellifera and Halictus sp. bees showed reduced visitation intensity after the application of deltamethrin or neem-based insecticides. Fungicide treatment alone did not influence the bee's visitation intensity. Deltamethrin-treated melon fields produced significantly lighter marketable fruits, and the melon yield was significantly lower in melon fields treated with the neem-based insecticide. Thus, our findings with such pollinator bees reinforce the idea that field applications of botanical pesticides may represent as risky as the applications of synthetic compounds, indicating that these alternative products should be submitted to risk assessments comparable to those required for synthetic products.

Neem oil based nanopesticide as an environmentally-friendly formulation for applications in sustainable agriculture: An ecotoxicological perspective.

Sustainable agriculture encourages practices that present low risks to the environment and human health. To this end, zein (corn protein) can be used to develop nanocarrier systems capable of improving the physicochemical properties of biopesticides, reducing their possible toxicity. Neem oil extracted from the Azadirachta indica tree contains many active ingredients including azadirachtin, which is the active ingredient in multiple commercially available biopesticides. In this study, we describe the preparation and characterization of neem oil-loaded zein nanoparticles, together with evaluation of their toxicity towards nontarget organisms, using Allium cepa, soil nitrogen cycle microbiota, and Caenorhabditis elegans aiming to achieve the safer by design strategy. The spherical nanoparticles showed an average diameter of 278 ±â€¯61.5 nm and a good stability during the experiments. In the toxicity assays with A. cepa, the neem oil-loaded zein nanoparticles mitigated the increase in the DNA relative damage index caused by the neem oil. Molecular genetic analysis of the soil nitrogen cycle microbiota revealed that neem oil-loaded zein nanoparticles did not change the number of genes which encode nitrogen-fixing enzymes and denitrifying enzymes. In C. elegans, the neem oil-loaded zein nanoparticles had no toxic effect, while neem oil interfered with pharyngeal pumping and GST-4 protein expression. These neem oil-loaded zein nanoparticles showed promising results in the toxicity studies, opening perspectives for its use in crop protection in organic agriculture.

Nanocapsules Containing Neem (Azadirachta Indica) Oil: Development, Characterization, And Toxicity Evaluation.

In this study, we prepared, characterized, and performed toxicity analyses of poly(ε-caprolactone) nanocapsules loaded with neem oil. Three formulations were prepared by the emulsion/solvent evaporation method. The nanocapsules showed a mean size distribution around 400 nm, with polydispersity below 0.2 and were stable for 120 days. Cytotoxicity and genotoxicity results showed an increase in toxicity of the oleic acid + neem formulations according to the amount of oleic acid used. The minimum inhibitory concentrations demonstrated that all the formulations containing neem oil were active. The nanocapsules containing neem oil did not affect the soil microbiota during 300 days of exposure compared to the control. Phytotoxicity studies indicated that NC_20 (200 mg of neem oil) did not affect the net photosynthesis and stomatal conductance of maize plants, whereas use of NC_10 (100:100 of neem:oleic acid) and NC_15 (150:50 of neem:oleic acid) led to negative effects on these physiological parameters. Hence, the use of oleic acid as a complement in the nanocapsules was not a good strategy, since the nanocapsules that only contained neem oil showed lower toxicity. These results demonstrate that evaluation of the toxicity of nanopesticides is essential for the development of environmentally friendly formulations intended for applications in agriculture.

Selectivity of plant extracts for Trichogramma pretiosum Riley (Hym.: Trichogrammatidae).

We evaluated the selectivity of three plant extracts with potential insecticidal effects for the parasitoid Trichogramma pretiosum Riley, which is commonly used in biological pest control. The plant extracts assayed were an acetone extract of Toona ciliata M. Roem., commercial neem oil, and a nanoencapsulated formulation of neem oil (NC40). The toxicity of the plant extracts to T. pretiosum was evaluated according to the recommendations of the International Organization for Biological Control- IOBC Working Group. We assessed the susceptibility of adults of the maternal and F1 generations and immature stages of T. pretiosum to the extracts. Females exposed to egg cards treated with commercial neem oil parasitized almost 70% fewer eggs than control eggs treated with water; and this extract was therefore classified as slightly harmful. When the eggs were offered to females 24h after treatment with neem oil and aqueous NC40, the parasitism rate also decreased, and the two extracts were classified as slightly harmful. Adult emergence was lower for parasitoids that fed on host eggs offered 24h after the treatment with the T. ciliata extract, which was considered slightly harmful. The emergence of T. pretiosum from eggs, larvae and pupae treated with the different plant extracts, did not decrease compared to development stages treated with the water control. The use of T. pretiosum, combined with the application of an ethanol extract of T. ciliata and a nanoencapsulated formulation of neem, appears to be feasible in view of these low toxicity indices.

Morphological alterations in the synganglion and integument of Rhipicephalus sanguineus ticks exposed to aqueous extracts of neem leaves (Azadirachta indica A. JUSS).

Currently, the necessity of controlling infestation by ticks, especially by Rhipicephalus sanguineus, has led researchers and public health managers around the world to search for new and more efficient control methods. This way, we can highlight neem (Azadirachta indica A. Juss) leaf, bark, and seed extracts, which have been very effective on tick control, and moreover causing less damage to the environment and to the host. This study showed the potential of neem as a control method for R. sanguineus through morphological and morphometric evaluation of the integument and synganglion of females, in semiengorged stage. To attain this, routine techniques of optical microscopy, scanning electron microscopy and morphometry of the cuticle and subcuticle of the integument were applied. Expressive morphological alterations were observed in both organs, presenting a dose-dependent effect. Integument epithelial cells and nerve cells of the synganglion showed signs of cell vacuolation, dilated intercellular boundaries, and cellular disorganization, alterations not previously reported in studies with neem. In addition, variations in subcuticle thickness were also observed. In general, the effects of neem are multiple, and affect the morphology and physiology of target animals in various ways. The results presented in this work are the first evidence of its effects in the coating and nervous system of ticks, thus allowing an indication of neem aqueous extracts as a potential control method of the brown dog tick and opening new perspectives on acaricide use.

Modulation of anxiety-like behavior by the endocannabinoid 2-arachidonoylglycerol (2-AG) in the dorsolateral periaqueductal gray.

Anandamide and 2-arachidonoylglycerol (2-AG) are the two main endocannabinoids, exerting their effects by activating type 1 (CB1r) and type 2 (CB2r) cannabinoid receptors. Anandamide inhibits anxiety-like responses through the activation of CB1r in certain brain regions, including the dorsolateral periaqueductal gray (dlPAG). 2-AG also attenuates anxiety-like responses, although the neuroanatomical sites for these effects remained unclear. Here, we tested the hypothesis that enhancing 2-AG signaling in the dlPAG would induce anxiolytic-like effects. The mechanisms involved were also investigated. Male Wistar rats received intra-dlPAG injections of 2-AG, URB602 (inhibitor of the 2-AG hydrolyzing enzyme, mono-acylglycerol lipase--MGL), AM251 (CB1r antagonist) and AM630 (CB2r antagonist). The behavior was analyzed in the elevated plus maze after the following treatments. Exp. 1: vehicle (veh) or 2-AG (5 pmol, 50 pmol, and 500 pmol). Exp. 2: veh or URB602 (30 pmol, 100 pmol or 300 pmol). Exp. 3: veh or AM251 (100 pmol) followed by veh or 2-AG (50 pmol). Exp. 4: veh or AM630 (1000 pmol) followed by veh or 2-AG. Exp. 5: veh or AM251 followed by veh or URB602 (100 pmol). Exp. 6: veh or AM630 followed by veh or URB602. 2-AG (50 pmol) and URB602 (100 pmol) significantly increased the exploration of the open arms of the apparatus, indicating an anxiolytic-like effect. These behavioral responses were prevented by CB1r (AM251) or CB2r (AM630) antagonists. Our results showed that the augmentation of 2-AG levels in the dlPAG induces anxiolytic-like effects. The mechanism seems to involve both CB1r and CB2r receptors.

Toxicity of Neem's oil, a potential biocide against the invasive mussel Limnoperna fortunei (Dunker 1857).

The golden mussel Limnoperna fortunei (Dunker 1857) is one of the most distributed Nuisance Invasive Species (NIS) in South America, and a threat of great concern for the industry of the area. In this study, we carried out toxicity tests made with a Neem's oil solution with L. fortunei larvae and benthonic adults (7, 13 and 19 ± 1 mm). Tests with non-target species (Daphnia magna, Lactuca sativa and Cnesterodon decemmculatus) were also made with the aim to evaluate the potential toxicity of the Neem's solution in the environment. The LC(100) of Neem's solution obtained for larvae was 500 µl/L, a value much higher than the one obtained for D. magna and C. decemmaculatus. Thus, we recommend that it should not be used in open waters. However, since the adults were killed in 72 h and the larvae in 24 h, this product can be used in closed systems, in man-made facilities.

Selective effects of natural and synthetic insecticides on mortality of Spodoptera frugiperda (Lepidoptera: Noctuidae) and its predator Eriopis connexa (Coleoptera: Coccinellidae).

Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is a serious pest of corn in several American countries. It is mainly controlled with synthetic insecticides. The objectives of this study were to evaluate the effects of the natural products, neem oil and pyroligneous extract, and the synthetic insecticide, lufenuron, at 2.50 mL water (0.25%) on the mortality of 2-, 4- and 6-day-old caterpillars of S. frugiperda, and their selectivities against fourth instar larvae of Eriopis connnexa Germar (Coleoptera: Coccinellidae). Four- and 6-day-old S. frugiperda caterpillars showed higher mortality after exposure to neem oil (83.33 +/- 0.83 and 89.58 +/- 0.90%, respectively) and lufenuron (95.83 +/- 0.96 and 85.41 +/- 0.83%), compared to pyroligneous extract (68.75 +/- 0.69 and 31.25 +/- 0.31%). The deleterious effect of pyroligneous extract was higher in 2- (83.33 +/- 0.83% mortality) and 4-day-old (68.75 +/- 0.69%) S. frugiperda caterpillars than in 6-day-old caterpillars (31.25 +/- 0.31%). Larval mortality of the predator E. connexa was lower with neem oil and pyroligneous extract (25.00 +/- 0.33%) than with lufenuron (91.66 +/- 1.22%). Neem oil is thus recommended for control of S. frugiperda because of its high toxicity, combined with its relatively low toxicity to larvae of the natural enemy E. connexa.