Avaliação da presença de fármacos em lodo de estação de tratamento de esgotos, antes e após processo de biorremediação por compostagem / Evaluation of the presence of drugs in sewage treatment station sludge before and after the bioremediation process by composting

Resumo A presença de fármacos residuais no ambiente é um problema crescente e de abrangência global, com ampla quantidade de pesquisas visando sua quantificação. Porém, seu comportamento e efeitos, tanto no ambiente quanto para o ser humano, ainda não são bem conhecidos. Neste trabalho foram avaliados sete fármacos: Atenolol, Carbamazepina, Clonazepan, Ibuprofeno, Paracetamol, Sinvastatina e Fluoxetina, em amostras de lodo proveniente de uma estação de tratamento de esgotos do sistema público. A extração dos compostos na matriz foi baseada na sua lixiviação em água e a quantificação analítica foi determinada por um sistema de cromatógrafo líquido de ultra performance, acoplado ao espectrômetro de massas. Os resultados indicam a possível movimentação de alguns fármacos da fase aquosa para a sólida. Observou-se que a compostagem do lodo pode favorecer a biodegradação ou mobilização dos compostos, pois houve redução significativa nas concentrações, quando comparados os valores obtidos antes e após o processo de compostagem. A não detecção de um composto não significa necessariamente que ele tenha sido degradado por microrganismos, entretanto, estudos pretéritos chegaram a resultados semelhantes, o que corrobora para a adequação da proposta metodológica e dos procedimentos adotados, contribuindo para a produção de resultados confiáveis.

Abstract The presence of residual drugs in the environment is a growing global issue, with many studies seeking to quantify it. However, its behavior and effects, both on the environment and on humans are still mostly unknown. In this study, seven drugs were evaluated: Atenolol, Carbamazepine, Clonazepam, Ibuprofen, Paracetamol, Simvastatin, and Fluoxetine, from samples of sludge from a sewage treatment plant of the public system. The extraction of the compounds from the matrix was based on the leaching of the compounds in water and the analytical quantification was determined by an ultra-performance liquid chromatograph system, coupled to the mass spectrometer. The data indicate the possible movement of some drugs from aqueous to solid phase. We observed that the composting of the sludge could favor the biodegradation or mobilization of the compounds since there was a significant reduction in the concentrations, when comparing the values obtained before and after the composting process. The non-detection of a compound does not necessarily mean that it has been fully degraded by microorganisms; however, past studies reached similar results, which corroborates the adequacy of the methodological proposal and the adopted procedures, contributing to the production of reliable results.

Performance of herbicides in sugarcane straw / Dinâmica de herbicidas em palha de cana-de-açúcar

The process of mechanical harvesting of sugarcane generates a large deposition of straw on the soil surface, providing a coverage that several studies have found important for reducing the weed population. Although such coverage reduces weed infestations, additional management, including chemical control, is still needed. Thus, this study aimed to evaluate the leaching of atrazine, pendimethalin, metribuzin, clomazone, diuron and hexazinone in sugarcane straw. The experiment was conducted at the School of Agronomic Engineering at UNESP (Sao Paulo State University) - Botucatu/SP. The sugarcane straw was collected in the field, cut and placed in quantities of 10t ha-1 in the capsules used as experimental units. The experimental design was completely randomized, using six herbicide treatments and four replications. Within 24 hours after the herbicides were applied in capsules with straw, five different rainfalls (5, 10, 20, 50 and 100mm) were simulated. The leached water was collected for chromatographic analysis. The herbicide percentages that crossed the straw layer were statistically correlated with the rainfall amount by the Mitscherlich model that compares the facility of herbicide removal from sugarcane straw. In summary, pendimethalin did not present quantified transposition of the product by sugarcane straw even with a rain simulation of 100 mm. Furthermore, two different profiles of facility to transpose the herbicides in straw were found: one for metribuzin and hexazinone that crossed quickly through the straw layer and another for atrazine, diuron and clomazone that required more rainfall to be leached from coverage to the soil according to the maximum removable amount of each herbicide.

O processo de colheita mecanizada de cana-de-açúcar gera uma grande quantidade de palhada depositada na superfície do solo e vários trabalhos destacam a importância da manutenção dessas coberturas em sistemas de cana crua, com destaque na redução da população de plantas daninhas. Mas, apesar de a cobertura vegetal reduzir a infestação das plantas daninhas, medidas complementares, como o controle químico, ainda são necessárias. Dessa forma, o objetivo deste estudo foi avaliar a transposição dos herbicidas atrazine, pendimethalin, metribuzin, clomazone, diuron e hexazinone em palha de cana-de-açúcar. O experimento foi desenvolvido na Faculdade de Ciências Agronômicas da UNESP - Botucatu/SP. A palha da cana-de-açúcar foi coletada em campo, cortada e colocada em quantidades de 10t ha-1 em recipientes plásticos utilizados como unidades experimentais. O delineamento experimental foi inteiramente casualisado, e foram testados seis tratamentos (atrazine, pendimethalin, metribuzin, clomazone, diuron e hexazinone) e quatro repetições. Nas cápsulas com palha e pulverizadas, foi realizado, no período de 24 horas após a aplicação, a simulação de diferentes lâminas de chuva (5, 10, 20, 50 e 100mm) e coletada a água lixiviada para posterior quantificação cromatográfica. Como análise estatística, os herbicidas foram correlacionados com as lâminas de chuva e foi usado o modelo de Mitscherlich para ajuste dos dados, que permitiu comparar a facilidade de remoção dos herbicidas pela palha. Em síntese, para o pendimethalin, não se observou transposição quantificada do produto pela palha de cana-de-açúcar, mesmo com uma lâmina de 100mm de chuva. Foram encontrados dois perfis diferentes de facilidade de transposição dos herbicidas em palha: um para o metribuzin e hexazinone, que passaram rapidamente pela camada de palha, e outro para atrazine, clomazone e diuron, que precisaram de uma quantidade superior de chuva para serem lixiviados da cobertura vegetal, em função da quantidade máxima removível dos herbicidas.

Detecção da tolerância de diferentes espécies de capim-colchão a herbicidas inibidores do fotossistema II utilizando a técnica da fluorescência / Detection of crabgrass species tolerance to photosystem II inhibitor herbicides using fluorescence technique

Este estudo teve como objetivo investigar a tolerância ou susceptibilidade das espécies de capim-colchão (Digitaria ciliaris, D. horizontalis e D. nuda) a herbicidas inibidores do fotossistema II (FSII), por meio da técnica da fluorescência, utilizando a taxa de transferência de elétrons do FSII como indicador. Para isso, foi conduzido um experimento em esquema fatorial 3x6, com três espécies de capim-colchão e seis tratamentos (ametryn, hexazinone, diuron+hexazinone, amicarbazone, diuron, tebuthiuron e testemunha) aplicados em pós-emergência. Foi monitorada a taxa de transporte de elétrons (ETR) do FSII em intervalos de tempo crescente e aferida a massa seca das plantas aos 21 dias após a aplicação. A partir dos dados da ETR, calculou-se um valor numérico representativo da taxa de inativação da ETR. Os resultados demonstraram que todas as espécies de capim-colchão estudadas são susceptíveis aos herbicidas ametryn e hexazinone (valores da taxa de inativação da ETR superiores a 10.000). Os herbicidas diuron e tebuthiuron apresentaram menores taxas de inativação da ETR para a D. nuda (3.585 e 3.497, respectivamente) e maiores para as espécies D. ciliaris e D. horizontalis (acima de 10.000), enquanto que o herbicida amicarbazone apresentou valor intermediário para D. nuda (7.967). O monitoramento da ETR foi eficiente para verificar a atuação dos herbicidas nas diferentes espécies estudadas.

This study aimed to investigate the tolerance or susceptibility of crabgrass species (Digitaria ciliaris, D. horizontalis and D. nuda) to herbicides that target photosystem II (PSII), by fluorescence technique using the electron transport rate (ETR) through PSII as an indicator. An experiment was conducted under a factorial arrange (3x6), with three species of crabgrass and six treatments (ametryn, hexazinone, diuron+hexazinone, amicarbazone, diuron, tebuthiuron and control) applied in post-emergence. The ETR through PSII was monitored over time and the plants dry weight measured at 21 days after application. A numerical value of the ETR inactivation was calculated from the data collected. The results showed that the three crabgrass species studied are susceptible to ametryn and hexazinone (ETR inactivation value higher than 10,000). Moreover, diuron and tebuthiuron provided lower ETR inactivation for D. nuda (3,585 and 3,497, respectively) and higher rates for D. horizontalis and D. ciliaris (more than 10,000), whereas amicarbazone showed intermediate inactivation rate for D. nuda (7,967). Monitoring the ETR showed to be an efficient form to verify the herbicides performance in the different species studied here.

Aplicação em subdose do haloxyfop-methyl na aveia-preta / Low doses of haloxyfop-methyl in black oat

Uma das limitações para sustentabilidade do sistema de cultivo em plantio direto é a rápida decomposição da matéria seca. Um dos mecanismos conhecidos para minimizar este processo e potencializar a permanência da palhada no solo é alterar as vias metabólicas dos polímeros de lignina com aplicação de subdoses de herbicida, interferindo diretamente na resistência à degradação dos restos vegetais pelos microrganismos. Com este objetivo, o herbicida Verdict*R (haloxyfop-methyl) foi aplicado em subdoses nas plantas de aveia-preta e o efeito correlacionado com a altura das plantas, produtividade da matéria seca e metabolismo de lignina. Ensaios preliminares em casa de vegetação foram realizados para determinar as faixas de subdoses a serem aplicadas no experimento definitivo em campo. Os resultados obtidos demonstraram aumento de 24% na produtividade de matéria seca dos experimentos conduzidos em casa de vegetação com a aplicação de 3,125g do ingrediente ativo de haloxyfop-methyl por hectare (i.a. ha-1) nesta subdose, não foram observadas alterações na altura e no metabolismo de lignina nas plantas de aveia-preta. No campo, a concentração de 2,5g i. a. ha-1 do haloxyfop-methyl já foi suficiente para reduzir em 9% a taxa de lignificação sem interferir na altura e produtividade das plantas, sendo este resultado favorável à velocidade de degradação da palhada no plantio direto.

One of the major constraints to sustainable of the tillage is the rapid decomposition of dry the matter. One of the mechanisms known to facilitate this process is the interference in pathways of polymers of lignin from tests of application of low doses of herbicide based on the relation that higher the content of the lignin greater the resistance to degradation. With this purpose, the herbicide Verdict * R (haloxyfop-methyl) was used to verify the effect of low doses at the height of the plants, productivity of dry matter and in the metabolism of the lignin in plants of black oat. Preliminary tests in the greenhouse were realized for adjustments ​​to the best low doses to be applied in the definitive experiment in the production area. The results obtained were 24% of increase in productivity in the experiments conducted in greenhouse, from the application of 3.125g of the active ingredient of the haloxyfop-methyl per hectare, in this subdose no changes was observed in growth and content of lignin in plants of black oat. In the field, the concentration of 2.5g i. a. ha-1 of haloxyfop-methyl decreased in 9% the lignification rate without interfering with the height and productivity of the plants, this being favorable to the degradation rate of stubble tillage on outcome.

Adjuvantes na deriva de 2,4-D + glyphosate em condições de campo / Adjuvants on spray drift of 2,4-D + glyphosate in field conditions

O objetivo do trabalho foi avaliar o efeito de adjuvantes sobre a deriva em aplicações da mistura de 2,4-D + glyphosate. O experimento foi conduzido em condições de campo em delineamento inteiramente casualizado. Os tratamentos corresponderam às soluções com a mistura dos herbicidas 2,4-D + glyphosate (670 e 1068g ha-1, respectivamente) adicionando-se os adjuvantes (v v-1): óleo mineral (0,5%); agente antideriva (0,09%); espalhante adesivo A (0,1%); fertilizante líquido (0,05%); espalhante adesivo B (0,25%); e somente os herbicidas (testemunha). Foram utilizados para determinação de deriva fios de náilon externos à área de aplicação (1, 5, 10, 20, 50, 100 e 200m de distância) com quatro repetições e seis cilindros de espuma posicionados sobre a barra do pulverizador para coleta de gotas sujeitas à deriva. As aplicações foram realizadas simultaneamente, utilizando-se um sal traçador específico em cada solução de aplicação para quantificar os depósitos por meio de espectrofotômetro. Não foi possível verificar efeito dos adjuvantes sobre a deriva nas diferentes distâncias da área de aplicação. Com base nas gotas coletadas acima da barra de pulverização, constatou-se que a suscetibilidade à deriva foi menor com o óleo mineral e o agente antideriva. O risco de deriva foi maior com o fertilizante líquido e o espalhante adesivo B.

The aim of the research was to evaluate the effect of adjuvants on the spray drift applications from mixture of 2,4-D + glyphosate. The trial was carried out in field conditions in a completely randomized design. The treatments corresponded to solutions containing mixture of the herbicides 2,4-D + glyphosate (670 and 1068g ha-1, respectively) adding the adjuvants (v v-1): mineral oil (0.5%); anti-drift agent (0.09%); spreader-sticker A (0.1%); liquid fertilizer (0.05%); spreader-sticker B (0.25%); and only herbicides without adjuvantes (control). Nylon strings were used to drift determination outside the application area (1, 5, 10, 20, 50, 100 and 200 m away) with 4 replications and six foam cylinders placed on the boom of the sprayer were used to collect the droplets subject to drift. The applications were performed simultaneously, using a specific salt tracer for each spray solution to quantify the deposits by spectrophotometer. It was not possible to verify effect of the adjuvants on drift at different distances of the application area. Based on droplets collected above the boom spray, it was found that susceptibility to drift was lower with the mineral oil and the anti-drift agent. The drift risk was higher with the liquid fertilizer and the spreader-sticker B.

Boron transport in Eucalyptus. 2. Identification in silico of a putative boron transporter for xylem loading in eucalypt

Boron (B) is a low mobility plant micronutrient whose molecular mechanisms of absorption and translocation are still controversial. Many factors are involved in tolerance to Boron excess or deficiency. Recently, the first protein linked to boron transport in biological systems, BOR1, was characterized in Arabidopsis thaliana. This protein is involved in boron xylem loading and is similar to bicarbonate transporters found in animals. There are indications that BOR1 is a member of a conserved protein family in plants. In this work, FORESTS database was used to identify sequences similar to this protein family, looking for a probable BOR1 homolog in eucalypt. We found five consensus sequences similar to BOR1; three of them were then used in multiple alignment analysis. Based on amino acid similarity and in silico expression patterns, a consensus sequence was identified as a candidate BOR1 homolog, helping deeper experimental assays that could identify the function of this protein family in Eucalyptus

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature, respectively. The alignment of EPSPS sequences allowed the identification of conserved regions that can be used to design specific primers for additional sequencings

Eucalyptus ESTs corresponding to the enzyme glutamine synthetase and the protein D1, sites of action of herbicides that cause oxidative stress

This work was aimed at locating Eucalyptus ESTs corresponding to the GS enzyme (Glutamine Synthetase, EC = 6.3.1.2) and to the D1 protein, which are directly related to resistance to herbicides that promote oxidative stress. Glutamine Synthetase corresponds to the site of action of the herbicide glufosinate. Herbicides that belong to groups such as ureas, uracils, triazines and triazinones act on the D1-Qb complex (receptor of electrons from the Photosystem II) by inactivating it. The clusters EGEQRT3302E01.g, EGEQRT3001F12.b; EGEZLV1203B04.g; EGBGFB1211H06.g and EGEZLV1205F09.g enclosed complete sequences (with 356 amino acids) of the Glutamine Synthetase enzyme. The cluster EGEQSL1054G06.g is a consensus of four reads and enclosed a complete sequence of D1 Protein (with 353 amino acids). The comparison of the sequences of Protein D1 from different species showed that the substitutions of serine (S) by glycine (G) or serine (S) by threonine (T) at the position 264 could produce plants resistant to herbicides that act on electron flow on Photosystem II. The sequence of amino acids corresponding to the cluster EGEQSL1054G06.g had a serine in position 264 indicating sensitivity of the Eucalyptus plants to herbicides that act on this site

Eucalyptus ESTs corresponding to the protoporphyrinogen IX oxidase enzyme related to the synthesis of heme, chlorophyll, and to the action of herbicides

This work was aimed at locating Eucalyptus ESTs corresponding to the PROTOX or PPO enzyme (Protoporphyrinogen IX oxidase, E.C. 1.3.3.4) directly related to resistance to herbicides that promote oxidative stress, changing the functionality of this enzyme. PROTOX, which is the site of action of diphenyl-ether (oxyfluorfen, lactofen, fomesafen), oxadiazole (oxadiazon and oxadiargyl), and aryl triazolinone (sulfentrazone and carfentrazone) herbicides, acts on the synthesis route of porphyrins which is associated with the production of chlorophyll a, catalases, and peroxidases. One cluster and one single read were located, with e-values better than e-70, associated to PROTOX. The alignment results between amino acid sequences indicated that this enzyme is adequately represented in the ESTs database of the FORESTs project