Isolamento e caracterização de bactérias degradadoras do herbicida diuron

Solos provenientes de áreas cultivadas com cana-de-açúcar, e histórico de utilização do herbicida diuron, foram utilizados para isolamento e caracterização de linhagens bacterianas com potencial de degradar este herbicida. As linhagens foram isoladas de solos da rizosfera de cana-de-açúcar e de solos das entre linhas. Noventa e oito linhagens foram isoladas, das quais 27 foram selecionadas para avaliação do crescimento em meio mineral sólido suplementado com diuron nas concentrações de 5, 10 e 20 mig mL-1. Neste experimento, 5 linhagens apresentaram um crescimento exponencial entre 0-48 horas (Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-CNPMA D12-12, D12-14 e D16-12) e entre 0-110 horas (Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-CNPMA D11-9 e D12-18), similar ao controle. A linhagem Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-CNPMA D12-12 identificada como Acinetobacter baumannii, foi selecionada para estudo de cinética de crescimento em meio mineral, com e sem fonte de nitrogênio, com e sem glicose (0,5%) e suplementado com diuron nas concentrações 20, 50 e 100 mig mL-1, apresentando variação em função da fonte de N, da presença da glicose e da concentração do diuron.

Seletividade de combinações de herbicidas latifolicidas com lactofen para a cultura de soja

Combinações de herbicidas visam o aumento do espectro de controle de plantas daninhas do complexo florístico, sendo usadas na agricultura com freqüência. Este trabalho teve como objetivo estudar a seletividade das combinações do herbicida lactofen, com bentazon, fomesafen, chlorimuron-ethyl e imazethapyr na cultura da soja, cultivar BR-37. Para isso, foi instalado um experimento de campo em Maringá-PR, em área livre de plantas daninhas evitando assim possíveis interferências sobre a cultura. Os tratamentos combinados foram obtidos aplicando-se de maneira contínua cada produto e suas doses a 75, 50 e 25% da dose comercial, seguindo as linhas de soja e novamente, aplicando-os perpendicularmente as mesmas, sendo assim, os locais onde houve o cruzamento e sobreposição das aplicações, formaram-se as combinações dos herbicidas, originando também, quatro testemunhas diagonais aos tratamentos. A comparação entre as médias de produtividade foi feita com o cálculo dos intervalos de confiança das testemunhas e de cada tratamento, adotando-se o teste t a 5%. As combinações foram seletivas para a cultura da soja. Verificaram-se reduções na produtividade de soja entre as combinações com os herbicidas nas maiores doses, sugerindo a necessidade de realizar novas pesquisas com os mesmos.

Droplet deposition during spray and leaf pH in aquatic weed control

A diversidade morfológica da superfície foliar existente entre as espécies de plantas e a presença de estruturas foliares como tricomas, estômatos, cutícula e ceras podem exercer grande influência na aderência e deposição das gotas de pulverização, assim como na absorção do herbicida. Desta forma, o objetivo do presente trabalho foi estudar, em quatro espécies de plantas daninhas aquáticas (Enhydra anagallis, Eichhornia crassipes, Heteranthera reniformis e Typha subulata), a área de molhamento de gotas de pulverização nas superfícies foliares adaxial e abaxial, bem como o pH foliar. O experimento foi realizado em Botucatu-SP. As plantas foram cultivadas em caixas d'água sob condições de campo, e quando atingiram seu pleno desenvolvimento (antes do florescimento), foram realizadas as avaliações de pH foliar e da área de molhamento de gotas de pulverização. As tensões superficiais das gotas aplicadas (0,5 mL), apresentadas pelas soluções de glyphosate aplicado isolado a 5,0% v v-1 (Rodeo 480 g L-1 e.a. produto comercial), glyphosate + Aterbane BR (5,0% + 0,5% v v-1), glyphosate + Silwet L-77 (5,0% + 0,05% v v-1), além das soluções com os adjuvantes isolados, Aterbane BR (0,5% v v-1) e Silwet L-77 (0,05% v v-1) foram respectivamente 72,1; 28,7; 23,3; 37,3 e 22,1 mN m-1. As médias obtidas de pH foliar variaram entre 5,50 e 7,50, destacando-se a espécie E. anagallis com valores de 6,68 e 7,02 para as faces adaxial e abaxial, respectivamente. Dentre as plantas daninhas aquáticas avaliadas T. subulata foi a espécie que apresentou as maiores médias de área de molhamento nas faces adaxial e abaxial da folha, proporcionada pelas as soluções de glyphosate + Aterbane BR, glyphosate + Silwet L-77 e Silwet L-77, com valores de 12,99-7,03; 20,04-17,95 e 31,81-25,91 mm², respectivamente.

ACUTE 2,4-DICHLOROPHENOXYACETIC ACID INTOXICATION IN CATTLE

The acute toxicity of 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in cattle. Steers were orally treated with 100, 300 or 600 mg 2,4-D/kg. Behavioral alterations, heart and respiratory functions, rectal temperature and ruminal movements were observed at 2, 4, 8, 12, 24, 48, 72 and 96 h after treatments. At these moments, blood and urine samples were collected and serum 2,4-D levels were determined. Results show that animals' vital functions and hematocrit were not modified by the herbicide. Other signs were dose and time-dependent and included motor alterations (weakness, lethargy, decreased general activity) and decreased ruminal movements and proteinuria. The herbicide was rapidly excreted and the intoxication signs were completely reversed. 2,4-D is an herbicide of small toxicological consequences for cattle kept under in natural grazing systems.

Determination of herbicides and a metabolite in human urine by liquid chromatography-electrospray ionization mass spectrometry

A method was developed to determine simazine, atrazine and their metabolite, 2-chloro-4,6-diamino-1,3,5-triazine, in urine. The presence of these herbicides in urine may reflect possible exposure to pesticides. Sample preparation involved protein precipitation and solid-phase extraction. The samples were analyzed by high-performance liquid chromatography-mass spectrometry. The detection limits were 0.4 mug/l and the analytes have a linear response in the interval 6-800 mug/l. The precision of the method was reflected in the RSD of <2.4% for the herbicides studied. Based on the detectable herbicide levels from spiked urine samples collected from unexposed volunteers, this method can be used to determine the low levels necessary for establishing reference values of the selected herbicides and the metabolite. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Rapid degradation of propanil in rice crop fields

Propanil and its major degradation product, 3,4-dichloroaniline (DCA), were monitored in surface water and soil samples from two rice fields of the Ebre Delta area (Tarragona, Spain) following agricultural application. On-line solid-phase extraction (SPE) (water) and Soxhlet extraction (soil) followed by liquid chromatography/diode array detection (LC/DAD) were used for the trace determination of both compounds. Unequivocal confirmation/identification was conducted by using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry, LC/APCI/MS (using negative and positive ionization modes). Concentrations of the herbicide propanil in water samples varied from 1.9 to 55.9 mu g/L. Propanil degraded very rapidly to DCA, and high concentrations of this product were found, varying from 16.5 to 470 mu g/L in water and 119 +/- 22 mu g/kg in soil samples. No detectable DCA (<0.001%) was found in the applied pesticide formulation, indicating that DCA formation took place after propanil application. These field results compared favorably with laboratory experiments showing that humic interactions had a strong influence on the pesticide degradation. The half-lifes under real conditions for propanil and DCA, calculated using a first-order decay, were 1.2 and 1.6 days, respectively.

Diuron lacks promoting potential in a rat liver bioassay

The promoting activity of the herbicide Diuron was evaluated in a medium-term rat liver carcinogenesis bioassay that uses as endpoint immunohistochemically identified glutathione S-transferase positive (GST-P+) foci. Male Wistar rats were allocated to the following groups: G1 to G6 were initiated for liver carcinogenesis by a single dose of diethylnitrosamine (DEN, 200 mg/kg) while groups G7 and G8 received only 0.9% NaCl (DEN vehicle). From the 2nd week animals were fed a basal diet (G1 and G7) or a diet added with Diuron at 125, 500, 1250, 2500 and 2500 ppm (G2 to G5 and G8, respectively) or 200 ppm Hexaclorobenzene (HCB; G6). The animals were submitted to 70% partial hepatectomy at the 3rd week and sacrificed at the 8th week. The herbicide did not alter ALT or creatinine serum levels. No conspicuous GST-P+ foci development was registered in non-initiated rats fed Diuron at 2500 ppm. While DEN-initiated animals fed Diuron at 1250 or 2500 ppm developed mild centrilobular hypertrophy, DEN-initiated HCB-fed animals showed severe liver centrilobular hypertrophy and significant GST-P+ foci development. These findings indicate that the medium-term assay adopted in this study does not reveal any liver carcinogenesis initiating or promoting potential of Diuron in the rat.

Multivariate analysis of photo-Fenton degradation of the herbicides tebuthiuron, diuron and 2,4-D

The degradation of herbicides in aqueous solution by photo-Fenton process using ferrioxalate complex (FeOx) as source of Fe2+ was evaluated under blacklight irradiation. The commercial products of the herbicides tebuthiuron, diuron and 2,4-D were used. The multivariate analysis, more precisely, the response surface methodology was applied to evaluate the role of FeOx and hydrogen peroxide concentrations as variables in the degradation process, and in particular, to define the concentration ranges that result in the most efficient degradation of the herbicides. The degradation process was evaluated by the determination of the remaining total organic carbon content (TOC), by monitoring the decrease of the concentrations of the original compounds using HPLC and by the chloride ion release in the case of diuron and 2,4-D. Under optimized conditions, 20min were sufficient to mineralize 93% of TOC from 2,4-D and 90% of diuron, including oxalate. Complete dechlorination of these compounds was achieved after 10 min reaction. It was found that the most recalcitrant herbicide is tebuthiuron, while diuron shows the highest degradability. However, under optimized conditions the initial concentration of tebuthiuron was reduced to less than 15%, while diuron and 2,4-D were reduced to around 2% after only 1 min reaction. Furthermore, it was observed that the ferrioxalate complex plays a more important role than H2O2 in the photodegradation of these herbicides in the ranges of concentrations investigated. (C) 2004 Elsevier Ltd. All rights reserved.

Influence of the iron source on the solar photo-Fenton degradation of different classes of organic compounds

In this work the influence of two different iron sources, Fe(NO3)(3) and complexed ferrioxalate (FeOx), on the degradation efficiency of 4-chlorophenol (4CP), malachite green, formaldehyde, dichloroacetic acid (DCA) and the commercial products of the herbicides diuron and tebuthiuron was studied. The oxidation of 4CP, DCA, diuron and tebuthiuron shows a strong dependence on the iron source. While the 4CP degradation is favored by the use of Fe(NO3)(3), the degradation of DCA and the herbicides diuron and tebuthiuron is most efficient when ferrioxalate is used. on the other hand, the degradation of malachite green and formaldehyde is not very influenced by the iron source showing only a slight improvement when ferrioxalate is used. In the case of formaldehyde, DCA, diuron and tebuthiuron, despite of the additional carbon introduced by the use of ferrioxalate, higher mineralization percentages were observed, confirming the beneficial effect of ferrioxalate on the degradation of these compounds. The degradation of tebuthiuron was studied in detail using a shallow pond type solar flow reactor of 4.5 L capacity and 4.5 cm solution depth. Solar irradiation of tebuthiuron at a flow rate of 9 L h(-1), in the presence of 10.0 mmol L-1 H2O2 and 1.0 mmol L-1 ferrioxalate resulted in complete conversion of this herbicide and 70% total organic carbon removal. (c) 2005 Elsevier Ltd. All rights reserved.

Determination of 2,4-dichlorophenoxyacetic acid and its major transformation product in soil samples by liquid chromatographic analysis

The 2,4-dichlorophenoxyacetic acid (2,4-D) is one of the most applied herbicides around the world to control broad leave herbs in many crops: In this study, a method was developed for simultaneous extraction and determination of 2,4D and its major transformation product, i.e., the 2,4-dichlorophenol (2,4-DCP), in soil samples. The herbicide and its degradation product were extracted twice from soil samples, after acidification, by dichloromethane on ultrasound system for 1 h. Both extracts were combined and filtrated in qualitative filter paper and Celitee. The total extract was concentrated in rotatory evaporator, dried under N-2 and finally dissolved in 1 ml of methanol. High Performance Liquid Chromatography with UV detection at 230 nm was used for analysis. Recoveries were obtained from soil samples fortified at 0.1, 1.0, 2.0, 3.0 and 4.0 mg kg(-1) levels and the results varied from 85 to 111% (for 2,4-D) and from 95 to 98% (for 2,4-DCP). For both compounds, the limits of quantification were 0.1 mg kg(-1), which were the loss level at which the accuracy and the precision were studied. Nevertheless, the limits of detection, calculated by considering the blank standard deviation and the minimum concentration level, were 0.03 and 0.02 mg kg for 2,4-D and 2,4-DCP, respectively. This proposed method was applied to soil samples of eucalyptus crops, which was previously treated with the herbicide. Despite that, neither 2,4-D nor its degradation product were detected 30 days after the herbicide application. (C) 2003 Elsevier B.V. B.V. All rights reserved.

EFFECTS OF 2,4-DICHLOROPHENOXYACETIC ACID (2,4-D) ON OPEN-FIELD BEHAVIOR AND NEUROCHEMICAL PARAMETERS OF RATS

The effects of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) on the central nervous system (CNS) were studied in rats. Behavioural and neurochemical studies were performed. Results show that acute and oral administration of dimethylamine 2,4-D was able to decrease locomotion and rearing frequencies and to increase immobility duration of rats observed in an open-field test. Treatment of rats with p-chlorophenylalanine (PCPA) was unable to change rat's open-field behaviour; 5-hydroxytryptophan (5-HTP) administration not only increased locomotion and rearing frequences but also decreased immobility duration. Pretreatment of the rats with PCPA and 5-HTP decreased and increased dimethylamine 2,4-D effects, respectively. The herbicide was not able to change the striatal levels of dopamine and homovanilic acid but decreased the striatal levels of serotonin (5-HT), as observed for the doses of 100 and 200 mg/kg and increased those of 5-hydroxyindoleacetic acid (5-HIAA) as measured after the 200 mg/kg dose treatment. When the levels of serotonin and 5-HIAA were measured at the brain stem level, only those of 5-HIAA were modified, being increased by diethylamine 2,4-D (60; 100 and 200 mg/kg); this increment on 5-HIAA levels was observed even 1 hr after pesticide administration. Further analysis showed that 2,4-D concentrations chromatographycally detected both in serum and brain of the intoxicated animals were dose-dependent, being found as early as 1 hr after the smaller dose of the herbicide used (10 mg/kg). The results suggest that diethylamine 2,4-D modify 5-HT functional activity within the CNS. Thus, the effects of the herbicide on open-field behaviour of rats could be attributed to a direct or indirect pesticide action on serotoninergic systems.

Multiresidue analysis of pesticides in water from rice cultures by on-line solid phase extraction followed by LC-DAD

An automated on-line solid phase extraction procedure followed by liquid chromatography with diode array detection was investigated for the determination of different classes of pesticides in water samples containing varied amount of humic substances. The different pesticides used were: carbendazin, carbofuran, atrazine, diuron, propanil, molinate, alachlor, parathion-ethyl, diazinon, trifluralin and the degradation products deisopropylatrazine and deethylatrazine. Humic substances extracted from a Brazilian sediment were used from 5 to 80 mg/l and their influence on recoveries was evaluated in neutral and acidic media. Recoveries higher than 70% were obtained for all the pesticides, from the preconcentration of 75 mi of aqueous sample fortified at 2 ng/ml using precolumns packed with PLRP-S. Good recoveries were obtained at neutral pH for most of the analytes up to 40 mg/l of humic acid. Only at 80 mg/l the recoveries were significantly affected, both at acidic and neutral pH. The method was applied to the determination of pesticides in river water spiked at 0.1 to 1 ng/ml. Detection limits obtained for water containing 10 mg/l of humic acid were between 0.05 and 0.3 ng/ml.

Variable dose rate application of herbicides using optical sensors

The aim of this study was to develop and evaluate a variable dose rate application of herbicides using an online electronic control based system with optical sensors for weed detection in forested areas. The proposed concept was to apply a basic dose on 100% of the area (aiming to control small weeds) and to apply a complementary patch-spraying dose only on areas with higher weed infestation. For that purpose, a conventional spray boom was adjusted to apply 40% of the herbicide dose on the full area and the optical sensors were used to control the application of the complementary dose (60%) only on areas with higher infestation. The results showed that the system performed adequately. Field applications presented herbicide savings around 20 to 30%, with a similar weed control performance as compared to the full dose application on 100% of the area.

Detection of Sourgrass (Digitaria insularis) Biotypes Resistant to Glyphosate in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Resposta de plantas daninhas ao óleo fúsel aplicado em pós-emergência inicial e tardia

This work aimed to study weed response to fusel oil rate applied at early and late post-emergence. The following species were studied at late and early post-emergence: Ipomoea hederifolia, Ipomoea quamoclit, Euphorbia heterophylla, Digitaria spp., Cenchrus echinatus and Panicum maximum.. The experiment was arranged in a completely randomized design with six treatments and four replications. Fusel oil, an alcohol distillation byproduct, was applied at rates of 50, 125, 250, 375 and 500 L ha(-1), plus control without application. The plots were constituted by polyethylene containers with 3 L capacity, filled with topsoil land from a fallow area. Visual symptoms of intoxication were verified at 7 and 30 days after application (DAA) and dry weight at 30 DAA. The variables were submitted to variance analysis according to the design, adjusting the data to obtain the dose-response curve by polynomial regression. The studied species were susceptible only to the application of 500 L ha(-1) of fusel oil at early or late plant post-emergence. The species Digitaria spp. was susceptible and E. heterophylla tolerant to fusel oil applied at early post-emergence, while the other species were moderately tolerant. E. heterophylla was susceptible, Digitaria spp., C. echinatus and P. maximum moderately tolerant and I. hederifolia and I. quamoclit tolerant to fusel oil applied at late post-emergence.