鱼类微囊藻毒素分析的新方法。Astraloheros facetetes的体外暴露研究。

Innotec Pub Date : 2020-06-03 DOI:10.26461/20.03
Natalia Badagian Baharian, M. Letamendía, M. Pirez, D. Carnevia, Beatriz M. Brena
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引用次数: 0

摘要

乌拉圭及其地区产生微囊藻毒素的蓝藻水华发生率很高,这对人类和动物来说是一个非常高的风险。为了研究微囊藻毒素(MC)在动物中的影响和存在,使用简单、低成本的方法很重要。作为在鱼类中实现这些目标的第一种方法,对暴露于含有微囊藻(60和600µg MCs/L)的水华微囊藻(Castanetas)进行亚慢性生物测定(18天)。尽管没有死亡,但组织病理学显示肝脏脂肪浸润,这在暴露于最高浓度的鱼类中更为相关。为了分析组织中的MCs,优化了两种基于高度特异性重组骆驼抗体(纳米体)的敏感免疫化学方法:ELISA和使用功能化磁性颗粒的定量MALDI-TOF。这些方法是最近在当地开发的。良好的相关性ELISA/MALDI-TOF(r Spearman=0.988,p<10-7)突出了这种ELISA作为一种简单且具有成本效益的工具的潜力,可以最大限度地减少通过参考方法分析的样本。面星藻组织中MCs的浓度是相关的,与其他物种和自然界鱼类的生物测定一致。这突出了分析鱼类消费中MC的重要性。
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Nuevas metodologías para el análisis de microcistinas en peces. Estudio de Astraloheros Facetus expuestos in vitro.
The high incidence of cyanobacterial blooms producing microcystins in Uruguay and its region, represent a very high risk for humans and animals. To study the impact and the presence of microcystins (MCs) in animals, it is important to use simple, low-cost methods. As a first approach to these objectives in fish, a sub-chronic bioassay (18 days) was performed with Astraloheros facetus (Castanetas) exposed to a Microcystis spp, bloom containing MCs (60 and 600 µg MCs/L). Even though there was no mortality, the histopathology showed fatty infiltration of the liver, which was more relevant in fish exposed to the highest concentration. To analyze MCs in tissues, two sensitive immunochemical methods based on a highly specific recombinant llama antibody (nanobody) were optimized: ELISA and quantitative MALDI-TOF, using functionalized magnetic particles. The methods were recently developed locally. The excellent correlation ELISA/MALDI-TOF (r Spearman = 0.988, p <10-7), highlights the potential of this ELISA as a simple and cost-effective tool to minimize the samples to be analyzed by reference methods. The concentrations of MCs in tissue of Astraloheros facetus were relevant, consistent with bioassays in other species and fish from nature. This highlights the importance of analyzing MCs in fish for consumption.
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