Vischetti Costantino, Casucci Cristiano, De Bernardi Arianna, Monaci Elga, Tiano Luca, Marcheggiani Fabio, Ciani Maurizio, Comitini Francesca, Marini Enrica, Taskin Eren, Puglisi Edoardo
Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Ancona, Italy.
Department of Life and Environmental Sciences, Marche Polytechnic University, Ancona, Italy.
Front Microbiol. 2020 Aug 18;11:1892. doi: 10.3389/fmicb.2020.01892. eCollection 2020.
This review describes the researches performed in the last years to assess the impact of pesticide sub-lethal doses on soil microorganisms and non-target organisms in agricultural soil ecosystems. The overview was developed through the careful description and a critical analysis of three methodologies based on culture-independent approaches involving DNA extraction and sequencing (denaturing gradient gel electrophoresis, DGGE; next-generation sequencing, NGS) to characterize the microbial population and DNA damage assessment (comet assay) to determine the effect on soil invertebrates. The examination of the related published articles showed a continuous improvement of the possibility to detect the detrimental effect of the pesticides on soil microorganisms and non-target organisms at sub-lethal doses, i.e., doses which have no lethal effect on the organisms. Considering the overall critical discussion on microbial soil monitoring in the function of pesticide treatments, we can confirm the usefulness of PCR-DGGE as a screening technique to assess the genetic diversity of microbial communities. Nowadays, DGGE remains a preliminary technique to highlight rapidly the main differences in microbial community composition, which is able to give further information if coupled with culture-dependent microbiological approaches, while thorough assessments must be gained by high-throughput techniques such as NGS. The comet assay represents an elective technique for assessing genotoxicity in environmental biomonitoring, being mature after decades of implementation and widely used worldwide for its direct, simple, and affordable implementation. Nonetheless, in order to promote the consistency and reliability of results, regulatory bodies should provide guidelines on the optimal use of this tool, strongly indicating the most reliable indicators of DNA damage. This review may help the European Regulation Authority in deriving new ecotoxicological endpoints to be included in the Registration Procedure of new pesticides.
本综述描述了过去几年开展的研究，以评估农药亚致死剂量对农业土壤生态系统中土壤微生物和非靶标生物的影响。通过仔细描述和批判性分析三种基于非培养方法的技术编写了本综述，这些方法包括用于表征微生物种群的涉及DNA提取和测序的方法（变性梯度凝胶电泳，DGGE；下一代测序，NGS）以及用于确定对土壤无脊椎动物影响的DNA损伤评估（彗星试验）。对相关已发表文章的研究表明，检测农药亚致死剂量（即对生物体无致死作用的剂量）对土壤微生物和非靶标生物的有害影响的可能性在不断提高。考虑到关于农药处理作用下土壤微生物监测的总体批判性讨论，我们可以确认PCR-DGGE作为评估微生物群落遗传多样性的筛选技术的有用性。如今，DGGE仍然是一种初步技术，能够快速突出微生物群落组成的主要差异，如果与基于培养的微生物学方法相结合，它能够提供更多信息，而全面评估必须通过NGS等高通量技术来实现。彗星试验是环境生物监测中评估遗传毒性的一种选择技术，经过数十年的应用已成熟，因其直接、简单且成本低廉而在全球广泛使用。尽管如此，为了提高结果的一致性和可靠性，监管机构应提供关于该工具最佳使用的指南，明确指出最可靠的DNA损伤指标。本综述可能有助于欧洲监管机构得出新的生态毒理学终点，以纳入新农药的注册程序。
