Brazilian experience with the medium-term multi-organ bioassay: Scientific and regulatory developments

In 1996 the Brazilian Institute for the Environment (IBAMA) officially adopted a variation of the multiorgan initiation-promotion DMBDD bioassay as a valid source of evidence of the carcinogenic potential of pesticides. The protocol adopted by IBAMA was a modification of the one originally proposed by researchers led by Nobuyuki Ito, from the Nagoya City University Medical School. Among the modifications established in the Brazilian protocol were the use of both sexes of the outbreed Wistar strain of rats and two positive control test chemicals. The adoption of the modified DMBDD protocol was instrumental during the last decade for qualifying technical people and to spread knowledge on chemical carcinogenesis in Brazil.

Multiple minima hypersurfaces procedures for biomimetic ligands screening

In this work the interaction of the pesticide carbaryl with two groups of biomimetic ligands, peptides and MIPs was screened by multiple minima hypersurfaces (MMH) procedures, through the AM1 semiempirical method. Data related to the properties of the molecular association of the complex biomimetic ligand-pesticide were obtained and compared with another molecular modeling algorithm named Leapfrog, as included in the Sybyl software package, and experimental results from the literature, remarking good correlation between them. All important MMH program parameters (cells number, box size, conformers) were studied and optimized with the aim of getting the minimum computation time without losing the correlation with experimental data. The data demonstrated that MMH approach can be used as a fast biomimetic ligand screening tool for MIPs. In the case of peptides the computation time was not comparable with the molecular dynamics methods conventionally used for this approach. © 2011 Springer Science+Business Media B.V.

Geoinformação na gestão dos recursos hídricos em sub-bacia hidrográfica

By remote sensing, geodatabase digital processing, information and expeditions to Capivara's sub-basin, was possible to identify the changes in the landscape mainly the expansion of eucalyptus, sugar-cane, and orange trees, being the last two, mainly on the Periferic Depression of Basaltic Cuesta. A significant portion of the soil on this geological area is formed from sandstones, providing high permeability to them, making them important places to groundwater recharge areas as sensible to contamination by pesticides. Throughout last decade was observed that the native vegetation fragments stabilization, keeping a reason of 26.5% on the land used between 2000 and 2010. The pasture decrease being substituted by eucalyptus, sugar-cane and orange trees call attention for the changes in the agribusiness model demanded by the current economic and social necessity. Pasture decrease followed by erosions decrease on sub-basin is evidence that these two aspects are strongly related.

Effects of spray nozzles and spray volume on spodoptera frugiperda management and narrow row corn performance

The high demand of pesticides in the production systems makes the application technology one of the main alternatives to optimize the products efficiency. In this context, the study aimed to evaluate the effects of spray nozzles and spray volumes on spraying deposits, armyworm control and crop corn performance in narrow row sowing system. The experiment was carried out at experimental area of Sao Paulo State University, Campus of Botucatu/SP, Brazil, during the 2009/2010 agricultural season, in randomized blocks with factorial scheme (2x2+1) and four replications. It was tested two flat fan spray nozzles (with and without air induction) combined with two spray volumes (100 and 200 L ha-1) plus a control treatment. There was no influence of spray nozzles (without air induction) in the spray deposits levels on plants. However, the flat fan nozzle with air induction was more effective on fall armyworm, with 100% of control against 47.84% from other at 15 days after spraying. The increase in the spray volume promoted high spray deposits (415.4 and 388.6 μL g-1 dry mass for flat fan nozzle with and without air induction, respectively at V10 growth stage) and consequently, the highest spray volume (200 L-1) was more efficient in the fall armyworm suppression, with 100% of control. All the technologies tested showed lower plant injury from fall armyworm. The insecticide sprayed with different technologies did not affect the parameters of plant height and leaf area index. The corn productivity was directly related with control efficiency of fall armyworm. © 2012 Academic Journals Inc.

Diferenças regionais do cultivo de café no Brasil

Brazil is one of the major coffee producers in the world, because of this, the goal of this study was to assess the regional differences of coffee cultivation for the reference crops 2001/2002 and 2002/03 by means of a life cycle assessment (LCA) in order to generate detailed production inventory data as well as quantify the potential environmental impacts of this crop. All information considered in this study (use of water, fossil based energy, fertilizers, pesticides and correctives) was taken from data collected from the producing farms. Four Brazilian coffee producer regions located at the Southeastern region were evaluated: Sul de Minas Gerais and Cerrado Mineiro in Minas Gerais State, and Mogiana and Alta Paulista regions in São Paulo State. The data refer to a production of 25.2 million kg of green coffee. Depending on the considered region, the production of 1,000 kg of green coffee requires, on average, approx. 9,300 to 13,000 kg of total energy, 70 to 130 kg of diesel, 6,500 to 12,700 kg of process water, 270 to 340 kg of fertilizers (NPK), 2.0 to 13.0 kg of pesticides, 230 to 600 kg of correctives, and yield around 1,600 to 1,900 kg/ha. Despite 20% of the coffee growers showing a good environmental performance, i.e. consumption of pesticides, fertilizers and correctives lower than the regional averages, this study has also identified some farms that can probably reduce the amount of some inputs and enhance their environmental performance.

Espacialização das estimativas de contaminação de águas subterrâneas por Tebuthiuron

Improper use of pesticides can lead to significant environmental impacts such as, contamination of environmental compartments, being the aquatic compartments the most vulnerable. In this context, the spatialization of pesticides concentrations estimative in groundwater provides important insights for decision making in managing and monitoring pesticides use. This study aimed to spatialize estimatives of groundwater contamination by Tebuthiuron, from different irrigation depths in the Rio Pardo basin, Pardinho-SP, Brazil. The simulations were performed using the ARAquá computer system, considering 0mm, 200 mm and 400 mm annual irrigation depths. Geostatistical techniques were used to obtain the spatial distribution of the simulated estimative. Tebuthiuron maps estimating concentration in groundwater were obtained by Kriging interpolation method, and indicated the areas with high potential for groundwater contamination. Considering all the simulations, it was concluded that there was no risk of groundwater contamination by Tebuthiuron in the study area.

Development of a support vector machine-based navigation system for driving a mobile robot through paths in plantations

The use of mobile robots turns out to be interesting in activities where the action of human specialist is difficult or dangerous. Mobile robots are often used for the exploration in areas of difficult access, such as rescue operations and space missions, to avoid human experts exposition to risky situations. Mobile robots are also used in agriculture for planting tasks as well as for keeping the application of pesticides within minimal amounts to mitigate environmental pollution. In this paper we present the development of a system to control the navigation of an autonomous mobile robot through tracks in plantations. Track images are used to control robot direction by preprocessing them to extract image features. Such features are then submitted to a support vector machine in order to find out the most appropriate route. The overall goal of the project to which this work is connected is to develop a real time robot control system to be embedded into a hardware platform. In this paper we report the software implementation of a support vector machine, which so far presented around 93% accuracy in predicting the appropriate route. © 2012 IEEE.

In vitro and in vivo inhibition of acetylcholinesterase and carboxylesterase by metals in zebrafish (Danio rerio)

Metals are natural components in ecosystems; however, if these elements are in excess, they can have adverse effects on living organisms. This study analyzes the interference of copper, lead, iron and cadmium in acetylcholinesterase (AChE) and carboxylesterase (CbE) activities in zebrafish. AChE was significantly inhibited in vitro by copper, iron, lead and cadmium at higher concentrations (10 and 20 mmol/L), whereas CbE was inhibited only at a concentration of 20 mmol/L. In vivo, only lead and cadmium were able to cause AChE inhibition at higher concentrations, while iron didn't cause any changes, and copper promoted an increase in AChE activity at a concentration of 0.06 mg/L. CbE activity did not change at any of the times (two and seven days) and concentrations tested, except in the case of copper exposure, which resulted in a decrease in CbE activity. Indeed, iodoacetamide treatment didn't changed AChE neither CbE activities, results which indicate that the metal inhibiting effect is probably not due to its biding to thiol groups close the active site of the enzyme. This outcome reveals that metals are important esterase inhibitors in zebrafish, and should be considered in environmental monitoring studies that use esterase inhibition as exposure biomarkers of organophosphate and carbamate pesticides. © 2012 Elsevier Ltd. All rights reserved.

Analysis of total glucosinolates and chromatographically purified benzylglucosinolate in organic and conventional vegetables

The limited availability of foods that are free of pesticides has led Brazil to search for alternative production methods to meet the desires of consumers. Currently, organic cultivation represents a production system that complies with general expectations of producers and consumers. Organic cultivation is particularly interesting mainly because of its effect on plant secondary metabolite content, which may help plants to naturally combat pests; in humans, these substances can also contribute to the prevention of chronic diseases. We report on the extraction of glucosinolates (both as total glucosinolates and as benzylglucosinolate) with trifluoroacetic acid addition in a 70:30 MeOH:water (v/v). Total glucosinolates, determined by a thioglucosidase coupled assay, were measured in different Brassicaceae species and were similar to values reported in the literature. For broccoli, analyses were carried out separately on inflorescences, leaves and stalks; analyses were also conducted on thermally processed samples to simulate cooking. Furthermore, when the analysis was conducted on conventional and organic products, the highest concentrations of these substances were most often found in organically cultivated Brassicaceae. The benzylglucosinolate concentrations were evaluated on the same samples using HPLC. The concentration of benzylglucosinolate was significantly higher in organically cultivated vegetables, as well. © 2012 Elsevier Ltd.

Chlorine disinfection of dye wastewater: Implications for a commercial azo dye mixture

Azo dyes, the most widely used family of synthetic dyes, are often employed as colorants in areas such as textiles, plastics, foods/drugs/cosmetics, and electronics. Following their use in industrial applications, azo dyes have been found in effluents and various receiving waters. Chemical treatment of effluents containing azo dyes includes disinfection using chlorine, which can generate compounds of varying eco/genotoxicity. Among the widely known commercial azo dyes for synthetic fibers is C.I. Disperse Red 1. While this dye is known to exist as a complex mixture, reports of eco/genotoxicity involve the purified form. Bearing in mind the potential for adverse synergistic effects arising from exposures to chemical mixtures, the aim of the present study was to characterize the components of commercial Disperse Red 1 and its chlorine-mediated decoloration products and to evaluate their ecotoxicity and mutagenicity. In conducting the present study, Disperse Red 1 was treated with chlorine gas, and the solution obtained was analyzed with the aid of LC-ESI-MS/MS to identify the components present, and then evaluated for ecotoxicity and mutagenicity, using Daphnia similis and Salmonella/microsome assays, respectively. The results of this study indicated that chlorination of Disperse Red 1 produced four chlorinated aromatic compounds as the main products and that the degradation products were more ecotoxic than the parent dye. These results suggest that a disinfection process using chlorine should be avoided for effluents containing hydrophobic azo dyes such commercial Disperse Red 1. © 2012 Elsevier B.V..

Influência de pontas de pulverização e adjuvantes no potencial de redução de deriva em túnel de vento

The drift is intimately linked to inappropriate use of pesticides and an important factor for reducing it, is the correct selection of spray nozzles and adjuvants. The objective of this work was to evaluate the drift potential in wind tunnel with different spray nozzles and different concentrations of adjuvants. The experiment was composed by six spray solutions (vegetable oil (in three concentrations), mineral oil, surfactant and reducing drift), which were applied with two nozzles, one pre-orifice flat fan (DG 8003 VS) and other with air induction (AI 8003 VS), totaling 12 treatments, with three repetitions. The equipment used was a wind tunnel, where the drift collections were made at different points. The treatments averages were compared using Confidence Interval at 5% probability. The analysis of the percentage of drift showed that the treatments had different behaviors. The nonylphenol ethoxylate adjuvant presented the highest drift when applied with the nozzle of pre-orifice and the lowest drift when applied with the air induction. The behavior of these nozzles when the oil-based adjuvant was used showed apposite results to those obtained for the surfactant. For the DG nozzles the lowest percentage of drift, at all analyzed distances, was observed to the treatment with vegetable oil (1.0%) and with the AI nozzles the lower drift was found for the treatment with nonylphenol ethoxylate (0.0625%), for the four distances in the collection. The result showed that both the spray nozzles and adjuvants alter directly the drift potential. There was no proportionately between the concentration of the oil-based adjuvant and the drift percentage.

Side-effects of thiamethoxam on the brain andmidgut of the africanized honeybee Apis mellifera (Hymenopptera: Apidae)

The development of agricultural activities coincides with the increased use of pesticides to control pests, which can also be harmful to nontarget insects such as bees. Thus, the goal of this work was assess the toxic effects of thiamethoxam on newly emerged worker bees of Apis mellifera (africanized honeybee-AHB). Initially, we determined that the lethal concentration 50 (LC50) of thiamethoxam was 4.28 ng a.i./μL of diet. To determine the lethal time 50 (LT50), a survival assay was conducted using diets containing sublethal doses of thiamethoxam equal to 1/10 and 1/100 of the LC50. The group of bees exposed to 1/10 of the LC50 had a 41.2% reduction of lifespan. When AHB samples were analyzed by morphological technique we found the presence of condensed cells in the mushroom bodies and optical lobes in exposed honeybees. Through Xylidine Ponceau technique, we found cells which stained more intensely in groups exposed to thiamethoxam. The digestive and regenerative cells of the midgut from exposed bees also showed morphological and histochemical alterations, like cytoplasm vacuolization, increased apocrine secretion and increased cell elimination. Thus, intoxication with a sublethal doses of thiamethoxam can cause impairment in the brain and midgut of AHB and contribute to the honeybee lifespan reduction. © 2013 Wiley Periodicals, Inc.

Dispersion and evaporation of droplets amended with adjuvants on soybeans

Quantification of the effects of adjuvants on droplet behaviour on plant surfaces is needed to improve pesticide spray application efficiency for soybeans. Dispersion and evaporation of single 300-μm diameter droplets amended with each of four spray adjuvants at five concentrations were investigated for four soybean plant surfaces (abaxial and adaxial leaflet surfaces, petiole, basal stem). The four adjuvants were a crop oil concentrate (COC), a modified seed oil (MSO), a non-ionic surfactant (NIS) and an oil surfactant blend (OSB). A single-droplet generator was used to produce and deposit 300-μm diameter droplets on target surfaces under controlled environmental conditions. Adjuvants significantly increased the dispersion (or wetted area) of droplets on plant surfaces. Droplet-wetted areas increased with increased adjuvant concentrations but not in direct proportion. The average increases of wetted areas across the four soybean plant surfaces were 443, 462, 416, or 343% when the spray mixture was amended with COC, MSO, NIS or OSB at the manufacturer-recommended concentrations, respectively. Among the four surfaces, the largest wetted area was on the abaxial surface, followed by the adaxial surface, the petiole and then the basal stem. Droplet evaporation times were inversely proportional to the wetted areas. The evaporation time of 300-μm diameter droplets ranged from 36 to 142. s on the four surfaces when the spray mixture was amended with an adjuvant, whereas the water-only droplets ranged from 161 to 190. s. The results demonstrated that use of adjuvants offers great potential to improve the homogeneity of sprayed pesticides, to increase spray coverage and to reduce pesticide application rates on soybean plants. These effects could benefit farmers economically and reduce environmental contamination by pesticides. © 2012.

Toxicological and histopathological effects of hydramethylnon on Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The leaf-cut ants are important agricultural pest, because they can cause intense defoliation in plants and destroy large areas cultivated. Although there are several works for the control of these insects by examining the toxicity of natural chemical compounds on various species of ants, few are focused on analyses of morphological changes caused in the affected organs. The aim of this study was to evaluate the effects of hydramethylnon on Atta sexdens rubropilosa workers through toxicological bioassays and morphological analysis of the post-pharyngeal glands, midgut, and Malpighian tubules of these ants. Hydramethylnon dissolved either in acetone (HA) or in a mixture of acetone and soy oil (HAO) was added to the artificial diet at a concentration of 200 μg/mL. The workers fed daily with the diet containing hydramethylnon showed higher mortality than the controls, especially when HAO was used. Moreover, light and electron microscopy revealed morphological alterations in the midgut and Malpighian tubules of workers treated with HA, whereas alterations of the post-pharyngeal glands were observed in the HAO-treated group. These results indicated that the presence of soy oil provided an alternate route for the ingestion of the formicide's active ingredient and corroborated previous studies that suggested a role for the post-pharyngeal glands in lipid metabolism. Our findings suggest that the oil may carry hydramethylnon to the gland lumen, resulting in lower quantity of the active ingredient in the intestinal lumen and Malpighian tubules that explains the lower degree of morphological alterations in these structures in the workers treated with HAO. These results may provide insight into the toxicological effects of hydramethylnon on leaf-cutting ants and the use of vegetable oil as an adjuvant in baits to control ants. © 2012 Elsevier Ltd.

Cellular responses in the Malpighian tubules of Scaptotrigona postica (Latreille, 1807) exposed to low doses of fipronil and boric acid

Studies of sub-lethal effects of pesticide residues on stingless bees are scarce and morphological analysis of organs would add information to toxicological analysis in order to clarify the continuous exposure of Scaptotrigona postica to insecticides. The aim of this study was to evaluate the morphology and histochemistry of the Malpighian tubules (excretory organ) of S. postica exposed to fipronil or boric acid to detect cellular responses that indicate toxicity or adaptative mechanisms to stress induced by exposure of worker bees to low doses of these chemical compounds. Newly emerged bees were submitted to toxicological bioassays and morphological analyses by optical microscopy and Transmission Electron Microscopy, as well as histochemical methods, were performed to detect proteins and glycoconjugates. Additionally, immunohistochemical detection of DNA fragmentation and HSP70 (70-kDa Heat shock protein) were performed to detect cell death and stress response, respectively. Statistical analysis, for the bioassays conducted with ingestion of contaminated diet with boric acid at 0.75% (w/w) or with fipronil at 0.1μg/kg of food, showed that the survival of bees that ingested the contaminated diets were significantly different to the survival rate presented by the control group (P<0.0001). Although some characteristics indicative of initiation of cell death were observed, the cells remained metabolically active in the processes of excretion and inactivation of chemical compounds. The data from this study reinforce the importance of research on sublethal effects of low doses of pesticides on bees in an attempt to assess a possible realistic dose and evaluate the risk assessment of stingless bee S. postica foraging in the vicinity of cultivated fields and/or in green urban areas. © 2013 Elsevier Ltd.