ABSTRACT
The triazine herbicide simazine is a pesticide commonly detected in surface and ground waters, although banned in most European countries since 2004. Concerns for humans and animal health result from its potential endocrine disrupting action, that can lead to reproductive disorders. The present in vitro study was undertaken to study simazine effects on swine granulosa cell function, namely cell viability, proliferation, steroidogenesis and NO production. Moreover, the ability of this substance to interfere with the angiogenetic process, a crucial event in reproductive function, was taken into account. Our data document that simazine treatment, at 0.1 or 10 μM concentration levels, stimulates granulosa cell proliferation and viability and impairs steroidogenesis, increasing in particular progesterone production. In addition, the in vitro angiogenesis bioassay revealed a significant simazine stimulatory effect on immortalized porcine Aortic Endothelial Cell proliferation. Collectively, these results show that simazine can display disruptive effects on ovarian cell functional parameters, possibly resulting in reproductive dysfunction. This hypothesis is also supported by the observed pro-angiogenetic properties of this herbicide, as already suggested for different endocrine disruptors.(AU)
Subject(s)
Animals , Swine/embryology , Simazine/adverse effects , Simazine/analysis , Granulosa Cells/chemistry , Swine/abnormalitiesABSTRACT
The triazine herbicide simazine is a pesticide commonly detected in surface and ground waters, although banned in most European countries since 2004. Concerns for humans and animal health result from its potential endocrine disrupting action, that can lead to reproductive disorders. The present in vitro study was undertaken to study simazine effects on swine granulosa cell function, namely cell viability, proliferation, steroidogenesis and NO production. Moreover, the ability of this substance to interfere with the angiogenetic process, a crucial event in reproductive function, was taken into account. Our data document that simazine treatment, at 0.1 or 10 μM concentration levels, stimulates granulosa cell proliferation and viability and impairs steroidogenesis, increasing in particular progesterone production. In addition, the in vitro angiogenesis bioassay revealed a significant simazine stimulatory effect on immortalized porcine Aortic Endothelial Cell proliferation. Collectively, these results show that simazine can display disruptive effects on ovarian cell functional parameters, possibly resulting in reproductive dysfunction. This hypothesis is also supported by the observed pro-angiogenetic properties of this herbicide, as already suggested for different endocrine disruptors.
Subject(s)
Animals , Granulosa Cells/chemistry , Simazine/analysis , Simazine/adverse effects , Swine/abnormalities , Swine/embryologyABSTRACT
The triazine herbicide simazine is a pesticide commonly detected in surface and ground waters, although banned in most European countries since 2004. Concerns for humans and animal health result from its potential endocrine disrupting action, that can lead to reproductive disorders. The present in vitro study was undertaken to study simazine effects on swine granulosa cell function, namely cell viability, proliferation, steroidogenesis and NO production. Moreover, the ability of this substance to interfere with the angiogenetic process, a crucial event in reproductive function, was taken into account. Our data document that simazine treatment, at 0.1 or 10 µM concentration levels, stimulates granulosa cell proliferation and viability and impairs steroidogenesis, increasing in particular progesterone production. In addition, the in vitro angiogenesis bioassay revealed a significant simazine stimulatory effect on immortalized porcine Aortic Endothelial Cell proliferation. Collectively, these results show that simazine can display disruptive effects on ovarian cell functional parameters, possibly resulting in reproductive dysfunction. This hypothesis is also supported by the observed pro-angiogenetic properties of this herbicide, as already suggested for different endocrine disruptors.
ABSTRACT
This paper describes the development of reversed phase sequential injection chromatography (SIC) methods for separation of simazine (SIM) and atrazine (AT), as well as their metabolites deethylatrazine (DEA), deisopropylatrazine (DIA) and hydroxyatrazine (HAT) exploiting silica based monolithic (50 × 4.6 mm) and core-shell (30 × 4.6 mm, 2.7 µm particles) columns. The separation was made by stepwise elution with two mobile phases: MP1 composed of 15:85 (v/v) acetonitrile: 2.5 mmol L(-1) acetic acid/ammonium acetate buffer (pH 4.2), and MP2, composed of 35:65 (v/v) acetonitrile: 2.5 mmol L(-1) acetic acid/ammonium acetate buffer (pH 4.2).The less hydrophobic compounds (DIA, HAT and DEA) eluted with MP1, whereas SIM and AT eluted with MP2. The method using core-shell column exhibited better chromatographic efficiency compared with monolithic column for separation of SIM and AT, but failed to provide base line separation of DIA and HAT. The proposed composition of mobile phases enabled the monolithic column to separate all the studied compounds with resolution >2.3 at flow rate of 35 µL s(-1) and sampling throughput of 8 analyses per hour, whereas in the core-shell the maximum flow rate allowed in the SIC system was 8 µL s(-1) (sampling throughput of 3 analyses per hour). The limits of detection were between 24 µg L(-1) (AT) and 40 µg L(-1) (DEA) using the monolithic column, and between 20 µg L(-1) (SIM) and 38 µg L(-1) (DEA) with the core-shell. Ultrasound-assisted extraction (80:20 v/v acetonitrile:water) of a soil sample enriched with the five triazines (250, 500 and 1000 µg kg(-1)) resulted recoveries between 51% and 121% of the spiked concentrations.
Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Reverse-Phase/methods , Flow Injection Analysis/methods , Herbicides/isolation & purification , Soil Pollutants/chemistry , Triazines/isolation & purification , Acetonitriles/chemistry , Chromatography, High Pressure Liquid/instrumentation , Chromatography, Reverse-Phase/instrumentation , Flow Injection Analysis/instrumentation , Herbicides/analysis , Triazines/analysisABSTRACT
A biorremediação é uma tecnologia que utiliza o metabolismo de microrganismos para eliminação ou redução, a níveis aceitáveis, de poluentes presentes no ambiente. Os herbicidas triazínicos são usados intensivamente no controle de ervas daninhas, principalmente na cultura de milho. Objetivou-se, neste trabalho, isolar fungos filamentosos de solos contaminados com herbicidas triazínicos (atrazine e simazine) e selecionar os microrganismos isolados quanto à capacidade de crescimento em meio adicionado de atrazine. Os microrganismos foram isolados, cultivados em meio Ágar-Batata-Dextrose (BDA) acidificado com ácido tartárico 10 por cento, adicionado de 50 mg.Kg-1 de atrazine e incubados por 5 dias a 25ºC. Foi realizada a medida diária do crescimento fúngico e calculada a velocidade de crescimento radial através de regressão linear dos raios das colônias utilizando-se a equação r(t) = a + VCR .t (r:raio; t: tempo; VCR: velocidade de crescimento radial). Os resultados de VCR foram analisados através de Anova simples e do teste de Tukey, para comparação de médias. Foram isolados 15 fungos, pertencentes aos gêneros Aspergillus, Penicillium e Trichoderma. As maiores VCRs foram obtidas com fungos Aspergillus (A1) e Penicillium (AS1), isolados de solo contaminado com atrazine e atrazine adicionado de simazine, respectivamente, que apresentaram VCRs de 1,57 mm.d-1 e 1,28 mm.d-1. O crescimento dos fungos em meio contaminado com a atrazine indica a possibilidade de utilização desses fungos em estudos de biorremediação de solos contaminados com herbicidas triazínicos.
Bioremediation is a technology that uses microrganism metabolism to quickly eliminate or reduce pollutants to acceptable levels into the environment. The triazine herbicides are intensively used to control harmful grass in the culture of maize. The aim of this work was to isolate filamentous fungi from soil contaminated with triazine herbicides and screening these fungi due to their ability of growth in a medium added by atrazine. The fungi were isolated, cultivated in potato-dextrose-agar plus 50 mg.Kg-1 of atrazine and incubated for 5 days at 25ºC. The measure of the rays of the colonies was carried out daily and the radial growth rate (RGR) through linear regression of colonies rays using the equation: r(t) = a + RGR .t (r:ray; t: time; RGR: radial growth rate). The RGR results were analyzed through analysis of variance and Tukey test for comparison of averages. Fifteen filamentous fungi from genus Aspergillus, Penicillium and Trichoderma were isolated. The highest RGRs were obtained with the fungi Aspergillus and Penicillium, when isolated from contaminated soil with atrazine and atrazine + simazine, respectively, showing RGR of 1.57 mm.d-1 and 1.28 mm.d-1. The growth of these fungi in atrazine contaminated meas indicates a possible of use of them in bioremediation experiments with contaminated soil containing triazine herbicides.