Early cytotoxic and genotoxic effects of atrazine on Wistar rat liver: A morphological, immunohistochemical, biochemical, and molecular study

Risk assessments suggest that intermediate and long-term exposure to triazine herbicides and its metabolites through water can cause severe damage to human health. The objective of this study was to investigate the possible effects of atrazine on Wistar rats submitted to subacute treatment. For this purpose, the activity of catalase and alanine aminotransferase was quantified, and the effect of the herbicide on cell membranes was examined based on the measurement of lipid peroxidation and consequent formation of malondialdehyde and on the mRNA expression of antioxidant enzymes (Mn-superoxide dismutase [SOD] and GSTM1) and connexins. In addition, we evaluated histopathological alterations in the liver, cellular expression of SOD and glutathione (GST), activation of heat shock proteins (HSPs) by immunohistochemistry, and the induction of apoptosis. The genotoxic potential of the herbicide was investigated by the micronucleus test in bone marrow smears. Adult male Wistar rats were treated with an aqueous solution of atrazine at a concentration of 400 mg/kg/day, by gavage, for 14 consecutive days. Control groups were also included. The results showed an increase of catalase levels and maintenance of the expression of antioxidant enzymes (SOD and GST). In addition, lipid peroxidation, hepatic tissue degeneration, activation of HSP90, increased levels of connexin mRNA, and genotoxicity were observed. In conclusion, atrazine induced early hepatic oxidative stress that triggered defense mechanisms to maintain the morphophysiological integrity of the liver. Further studies are needed to better understand the effects of this herbicide on human health. (C) 2011 Elsevier B.V. All rights reserved.

Tolerância de diferentes cultivares de cana-de-açúcar (Saccharum spp.) a herbicidas

This work aimed to evaluate the tolerance of sugarcane cultivars to sulfentrazone, imazapic, isoxaflutole, clomazone and ametryn + trifloxysulfuron-sodium. The experiment was arranged in a randomized block design in a split-plot scheme. The cultivars were allocated to the plots and the herbicides to the sub-plots (five 8.0 m long rows and 1.5 m spacing, with 4 repetitions. The herbicides sulfentrazone (0.8 kg ha(-1)), imazapic (0.147 kg ha(-1)), isoxaflutole (0.1125 kg ha(-1)), clomazone (1.1 kg ha(-1)), ametryn (1.463 kg ha(-1)) + trifloxysulfuron sodium (0.037 kg ha-1) and control were evaluated on 3-yr-old ratoons of the cultivars IACSP94-2094, IACSP94-2101, IACSP93-3046, IACSP94-4004, IAC86-2480 and RB72454 in post emergence. The traits evaluated were: plant toxicity symptoms in the plant leaves; total chlorophyll content and photochemical efficiency (Fv/Fm) at 15, 30 and 60 days after application (DAA); height (cm) at 30 and 270 DAA, and plant stand (stalk m(-1)) at 30 and 180 DAA. Diameter (cm), estimated productivity (t ha(-1)) and quality analysis were evaluated at 270 DAA. The sugarcane cultivars IACSP94-2094, IACSP93-3046, IACSP94-4004, IAC86-2480, RB72454, and IACSP94-2101 especially, were susceptible to clomazone up to 30 DAA, due to leaf chlorosis and lower chlorophyll content, but had no effect on quality characteristics and productivity. The cultivars were also tolerant to other herbicides.

Chlorophyll fluorescence as a marker for herbicide mechanisms of action

Photosynthesis is the single most important source of 02 and organic chemical energy necessary to support all non-autotrophic life forms. Plants compartmentalize this elaborate biochemical process within chloroplasts in order to safely harness the power of solar energy and convert it into usable chemical units. Stresses (biotic or abiotic) that challenge the integrity of the plant cell are likely to affect photosynthesis and alter chlorophyll fluorescence. A simple three-step assay was developed to test selected herbicides representative of the known herbicide mechanisms of action and a number of natural phytotoxins to determine their effect on photosynthesis as measured by chlorophyll fluorescence. The most active compounds were those interacting directly with photosynthesis (inhibitors of photosystem I and II), those inhibiting carotenoid synthesis, and those with mechanisms of action generating reactive oxygen species and lipid peroxidation (uncouplers and inhibitors of protoporphyrinogen oxidase). Other active compounds targeted lipids (very-long-chain fatty acid synthase and removal of cuticular waxes). Therefore, induced chlorophyll fluorescence is a good biomarker to help identify certain herbicide modes of action and their dependence on light for bioactivity. Published by Elsevier B.V.

Use of Biopolymeric Membranes for Adsorption of Paraquat Herbicide from Water

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Amicarbazone, a New Photosystem II Inhibitor

Amicarbazone is a new triazolinone herbicide with a broad spectrum of weed control. The phenotypic responses of sensitive plants exposed to amicarbazone include chlorosis, Stunted growth, tissue necrosis, and death. Its efficacy as both a foliar- and root-applied herbicide suggests that absorption and translocation of this compound is very rapid. This new herbicide is a potent inhibitor of photosynthetic electron transport, inducing chlorophyll fluorescence and interrupting oxygen evolution ostensibly via binding to the Q(B) domain of photosystem II (PSII) in a manner similar to the triazines and the triazinones classes of herbicides. As a result, its efficacy is susceptible to the most common form of resistance to PSII inhibitors. Nonetheless, amicarbazone has a good selectivity profile and is a more potent herbicide than atrazine, which enables its use at lower rates than those of traditional photosynthetic inhibitors.

Acute, subchronic and chronic 2,4-dichlorophenoxyacetic acid (2,4-D) intoxication in rats

The acute, subchronic and chronic toxicities of 2,4- dichlorophenoxyacetic acid (2,4-D) were studied in rats. Animals were exposed acutely (600 mg/kg), subchronically (200 ppm for 30 d) and chronically (200 ppm for 180 d) to 2,4-D by the oral route. Clinical, laboratory and histopathological methods were used as indicators of toxicity. After acute exposure, the herbicide decreased locomotor activity and induced ataxia, sedation, muscular weakness (mainly of the hind quarters) and gasping for breath; increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (AP), amylase activities and creatinine levels; decreased total protein (TP) and glucose levels; and increased hematocrit values. Subchronic and chronic 2,4-D exposures did not induce overt clinical signs or symptoms of intoxication. However, subchronic herbicide exposure increased AST activity and albumin and hematocrit values, and chronic exposure increased AST, AP and LDH activities, decreased amylase and glucose levels, but did not change hematocrit values. Chromatographic analysis of the serum of chronically exposed rats showed the presence of the herbicide; the amount found (3.76 ± 1.16 mg/ml) suggested the absence of 2,4-D accumulation within the body. Although macroscopic or histopathological lesions were not observed in acutely, subchronically or chronically 2,4-D exposed rats, the laboratory data obtained suggest tissue injuries after dosing, since the results are considered early indicators of primarily hepatic and muscle tissue damage.

Influência de herbicidas em macrófitas aquáticas

The effects of the herbicides atrazine, 2,4-D, glyphosate (direct and rodeo) and trifluralin were determined on the growth and death of fronds of the aquatic macrophytes Spirodela punctata (G.F.W. Meyer) Thompson and Salvinia minima Baker, through three series of concentrations of each herbicide in climatic chamber (3000 lux, 12/12 hours photoperiod, temperature 23 ± 1°C). After seven days, the concentrations of series 1 (0.025, 0.125, 0.625 and 3.125 mg.L -1) inhibited the growth of Salvinia minima. In Spirodela punctata, only the herbicides rodeo and trifluralin caused growth inhibition. These concentrations did not caused death of fronds in the species. The concentrations of series 2 (40, 60, 80 and 100 mg.L -1) of the herbicides 2,4-D, direct e trifluralin decreased growth of Salvinia minima; death of fronds was caused by 2,4-D and trifluralin to Salvinia minima, and by direct and trifluralin to Spirodela punctata. For the series 3 (200, 400, 600, 800 and 1000 mg.L -1), the herbicide 2,4-D caused growth decrease of both species. The herbicides rodeo and trifluralin showed to be the most lethal for both species. The herbicide atrazine caused death of fronds in Spirodela punctata.

Épocas de consórcio de alho com beterraba perante três manejos do mato sobre a produtividade e o lucro

The research was carried out with intercropped cultivation of garlic and beet, in Caçador, SC, Brazil. Four seeding epochs for beet (0, 15, 30 and 45 days after garlic planting) and three management systems for weeds (with herbicides, without control and with frequent weeding) were tested. It was hoped to determine the effects of this intercropping, in time and space, seeking the benefits in .weed control, efficient land use, productivity, commercial quality, and profitability. The randomized block design was in split-plots. The epochs represented the plots, and the management, the subplots, with four replicates. Napromide at 750 g/ha did not cause symptoms of intoxication in the garlic plants, nor in the beet. All the treatments in epochs at the initiation of intercropping, in three management systems for weeds, presented efficient land use values greater than 1. The profitability of the intercropping of garlic and beet only became evident in the management involving frequent weeding; in all epochs, profits were greater than for the respective epochs of monoculture of beet and monoculture of garlic.

Efficiency of safety measures applied to a manual knapsack sprayer for paraquat application to Maize (Zea mays L.)

The objectives of the present study were to evaluate the safety of mixer/loaders and applicators of paraquat to maize crop by knapsack sprayers and to determine the efficacy of safety measures applied to the sprayers. Potential dermal exposure (PDE) was evaluated in 22 worker body parts. The Cu2+ cation of a copper-based fungicide was used as tracer in the spray solution. Sanitary pads and cotton gloves were used to collect the pesticide solution on the sampled body parts. It was observed that paraquat application in front of the applicator's body (0.5 and 1.0 m lance) is unsafe because PDE was 1,979.8 ml/day (for 0.5 m lance) and 1,290.4 ml/day (for 1.0 m lance) and needs 50-80% and 37-69% control of PDE respectively. Control can be achieved by the use of protective garment on the legs and feet only, which received 92-93% of the PDE. Switching the spray nozzle to the back of the operator reduced the PDE by 98% and was sufficient to make working conditions safe, while maintaining the efficiency of application and making the work lighter and more comfortable. Mixer/loaders received 86% of the PDE to the hands and the work condition was safe (MOS > 1), however impermeable gloves could be used as a further safety measure.

Degradation of rice pesticides and their stability during storage of samples

Transformation of pesticides is directly related to the environmental conditions during application and transport of these compounds in the ecosystem. Rice fields include peculiar conditions, relatively high temperatures and wet conditions, leading to degradation processes, different from that observed in other agricultural systems. This article presents the degradation routes of some characteristic rice insecticides and herbicides under field conditions. A compilation of the pesticides that are usually applied during rice cultivation is included, with their main physico-chemical parameters. The stability of the pesticides by solid phase extraction systems during storage of rice samples is also discussed.

Períodos de interferência de erva-quente (Spermacoce atifolia) no crescimento inicial de eucalipto (Eucalyptus grandis)

The buttonweed (Spermacoce latifolia) is becoming a plant among the most current infesting eucalypts plantation in State of São Paulo due to the continual use of same herbicides and control methods. Owing this, this work aimed to evaluate the effects of periods of company and control of S. Iatifolia on the initial growth of Eucalyptus grandis, planted in winter and summer. Only one seedling of Eucalyptus was planted in amianthus cement box and submitted for crescent periods of company and control of S. Iatifolia (0, 20, 60 and 80 days in competition or not). The densities of plants of S. Iatifolia were 4 and 16 plants per m 2 (under winner and summer conditions). The experimental period was 100 days after the planting (DAP). The experimental design for both experiments was the completely randomized blocks (CRB) with ten treatments and four replicates. The results of plant high, stem diameter, dries weights and leaf area showed that the before interference period (BIP), whole period of prevention for interference (WPPI) and critical period of prevention for interference (CPPI) were 40, 60, and 60 DAR, respectively, under winner conditions. Under summer conditions, the WPPI and CPPI were 20, 80 and 20 to 80 DAR.

Molecular model of shikimate kinase from Mycobacterium tuberculosis

Tuberculosis (TB) resurged in the late 1980s and now kills approximately 3 million people a year. The reemergence of tuberculosis as a public health threat has created a need to develop new anti-mycobacterial agents. The shikimate pathway is an attractive target for herbicides and anti-microbial agents development because it is essential in algae, higher plants, bacteria, and fungi, but absent from mammals. Homologs to enzymes in the shikimate pathway have been identified in the genome sequence of Mycobacterium tuberculosis. Among them, the shikimate kinase I encoding gene (aroK) was proposed to be present by sequence homology. Accordingly, to pave the way for structural and functional efforts towards anti-mycobacterial agents development, here we describe the molecular modeling of M. tuberculosis shikimate kinase that should provide a structural framework on which the design of specific inhibitors may be based. © 2002 Elsevier Science (USA). All rights reserved.

Glutathione S-transferase activity in acetochlor, atrazine and oxyfluorfen metabolization in maize (Zea mays L.), sorghum (Sorghum bicolor L.) and wheat (Triticum aestivum L.) (Poaceae)

This experiment was conducted to evaluate the acetochlor, atrazine and oxyfluorfen herbicides plant selectivity, in relation to glutathione S-transferase activity (GST) in maize (Zea mays L.), sorghum (Sorghum bicolor L.) and wheat (Triticum aestivum L) (Poaceae) plants. GST activity was detected 24, 48 and 72 hours after treatment applications. The experiment's treatments consisted of spraying plants with water (control), acetochlor (3 L.ha -1), atrazine (4 L.ha -1) and oxyfluorfen (1 L.ha -1). The highest GST activities were observed in presence of acetochlor, mainly at 48 hours after treatment. These increments were 105, 148 and 118% when compared to maize, sorghum and wheat control groups, respectively. It is suggested that the GST may have a role in acetochlor degradation and it may be a reason for this herbicide's selectivity in these crops.