Comparison and optimization of biofilm DNA extraction from two types of soft microplastics in the coastal environment
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摘要: 微塑料因粒径小、比表面积大,可作为重金属、有机污染物以及病原微生物的载体。已有研究表明,微塑料表面附着的微生物主要以生物膜的形式存在。本研究以山东省海岸带环境中常见的两类软质塑料——发泡类聚苯乙烯(expanded polystyrene,EPS)和聚乙烯薄膜(polyethylene films,PE)为研究对象,比较了MP FastDNA®和MOBIO PowerSoil®两种DNA提取试剂盒对微塑料表面生物膜DNA的提取效果,探讨了不同的微塑料粒径和数量对DNA提取效果的影响。结果表明,MP FastDNA®试剂盒对两种软质微塑料表面生物膜DNA的提取浓度显著低于MOBIO PowerSoil®试剂盒(1.0~12.5倍)。采用MP FastDNA®试剂盒提取的PE表面DNA的浓度约为EPS的1.3~4.4倍。当微塑料数量不大于20片时,小粒径(1~3 mm)的EPS表面生物膜DNA浓度显著高于大粒径(3~5 mm) EPS,而对于PE薄膜则相反。对于两种粒径的EPS,微塑料表面DNA浓度均随着微塑料数量的增加而显著增加,但对于小粒径(1~3 mm)的PE薄膜,DNA浓度随微塑料数量的增加呈先增后减的趋势;而大粒径(3~5 mm)的PE薄膜表面DNA浓度随微塑料数量的增加而降低。微塑料的粒径和数量对其表面DNA提取效果影响的差异与微塑料的类型及其理化性质有关。本研究可为海洋与海岸环境中微塑料表面微生物群落组成与多样性研究提供方法支撑。
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关键词:
- 微塑料 /
- 生物膜 /
- DNA /
- 聚乙烯薄膜(PE) /
- 发泡类聚苯乙烯(EPS)
Abstract: Microplastics could serve as carriers for heavy metals, organic pollutants and pathogens in the coastal and marine environment due to their small sizes and large specific surface area. It has been reported that the microbial colonizers attaching on the surface of microplasti cs mainly assemblaged in the form of biofilms. In this study, two types of soft microplastics, expanded polystyrene (EPS) and polyethylene films (PE), were selected as tested microplastics that were commonly found in the coastal environment in Shandong province. Two DNA extraction kits (MP FastDNA® and MOBIO PowerSoil®) were used to compare their DNA extraction efficiency from different microplastic biofilms. Moreover, impacts from different particle sizes and quantities on DNA extraction efficiency of microplastic-associated biofilms were also evaluated. The results showed that concentrations of biofilm DNA extracted by FastDNA® kit were significantly higher than by MOBIO PowerSoil®kit (1.0-12.5 times). DNA concentrations from PE biofilm were about 1.3-4.4 times higher than that from EPS extracted by FastDNA®. When the quantities of microplastics were no more than 20, DNA concentrations from small sizes (1-3 mm) EPS were significantly lower than that from large sizes (3-5 mm), while it was totally in opposition for PE. With the increasing of microplastics quantities, DNA concentrations were significantly increased in both two sizes of EPS. But for small sizes (1-3 mm) of PE, DNA concentration showed an increasing initially and decreasing afterward trend with the increasing of PE quantities. However, for large sizes (3-5 mm) of PE, DNA concentrations were decreasing with the increasing of PE quantities. Different impacts of microplastic sizes and quantities on DNA concentrations could be attributed to the types and physicochemical properties of microplastics. Results from this study will be helpful for providing methodological support for the research of microbial community composition and diversity of microplastic-associated biofilms in the coastal and marine environment.-
Key words:
- microplastics /
- biofilms /
- DNA /
- polyethylene films (PE) /
- expanded polystyrene (EPS)
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Zhang Weiwei, Zhang Shoufeng, Wang Juying, et al. Microplastic pollution in the surface waters of the Bohai Sea, China[J]. Environmental Pollution, 2017, 231:541-548. Setälä O, Fleming-Lehtinen V, Lehtiniemi M. Ingestion and transfer of microplastics in the planktonic food web[J]. Environmental Pollution, 2014, 185:77-83. Li Jiana, Qu Xiaoyun, Su Lei, et al. Microplastics in mussels along the coastal waters of China[J]. Environmental Pollution, 2016, 214:177-184. Li Jiana, Yang Dongqi, Li Lan, et al. Microplastics in commercial bivalves from China[J]. Environmental Pollution, 2015, 207:190-195. Farrell P, Nelson K. Trophic level transfer of microplastic:Mytilus edulis (L.) to Carcinus maenas (L.)[J]. Environmental Pollution, 2013, 177:1-3. Brennecke D, Duarte B, Paiva F, et al. Microplastics as vector for heavy metal contamination from the marine environment[J]. Estuarine, Coastal and Shelf Science, 2016, 178:189-195. Mendoza L M R, Jones P R. Characterisation of microplastics and toxic chemicals extracted from microplastic samples from the North Pacific Gyre[J]. Environmental Chemistry, 2015, 12(5):611-617. Zhang Haibo, Zhou Qian, Xie Zhiyong, et al. Occurrences of organophosphorus esters and phthalates in the microplastics from the coastal beaches in north China[J]. Science of the Total Environment, 2018,616-617:1505-1512. Masó M, Garcés E, Pagès F, et al. Drifting plastic debris as a potential vector for dispersing Harmful Algal Bloom (HAB) species[J]. Scientia Marina, 2003, 67(1):107-111. Zettler E R, Mincer T J, Amaral-Zettler L A. Life in the "plastisphere":microbial communities on plastic marine debris[J]. Environmental Science & Technology, 2013, 47(13):7137-7146. McCormick A, Hoellein T J, Mason S A, et al. Microplastic is an abundant and distinct microbial habitat in an urban river[J]. Environmental Science & Technology, 2014, 48(20):11863-11871. Jiang Peilin, Zhao Shiye, Zhu Lixin, et al. Microplastic-associated bacterial assemblages in the intertidal zone of the Yangtze Estuary[J]. Science of the Total Environment, 2018, 624:48-54. Reisser J, Shaw J, Hallegraeff G, et al. Millimeter-sized marine plastics:a new pelagic habitat for microorganisms and invertebrates[J]. PLoS One, 2014, 9(6):e100289. Oberbeckmann S, Löder M G J, Labrenz M. Marine microplastic-associated biofilms-a review[J]. Environmental Chemistry, 2015, 12(5):551-562. Rummel C D, Jahnke A, Gorokhova E, et al. Impacts of biofilm formation on the fate and potential effects of microplastic in the aquatic environment[J]. Environmental Science & Technology Letters, 2017, 4(7):258-267. Kooi M, Van Nes E H, Scheffer M, et al. Ups and downs in the ocean:effects of biofouling on vertical transport of microplastics[J]. Environmental Science & Technology, 2017, 51(14):7963-7971. Zhang H. Transport of microplastics in coastal seas[J]. Estuarine, Coastal and Shelf Science, 2017, 199:74-86. Carpenter E J, Anderson S J, Harvey G R, et al. Polystyrene spherules in coastal waters[J]. Science, 1972, 178(4062):749-750. Lobelle D, Cunliffe M. Early microbial biofilm formation on marine plastic debris[J]. Marine Pollution Bulletin, 2011, 62(1):197-200. Webb H K, Crawford R J, Sawabe T, et al. Poly (ethylene terephthalate) polymer surfaces as a substrate for bacterial attachment and biofilm formation[J]. Microbes and Environments, 2009, 24(1):39-42. Oberbeckmann S, Loeder M G J, Gerdts G, et al. Spatial and seasonal variation in diversity and structure of microbial biofilms on marine plastics in Northern European waters[J]. FEMS Microbiology Ecology, 2014, 90(2):478-492. Carson H S, Nerheim M S, Carroll K A, et al. The plastic-associated microorganisms of the North Pacific Gyre[J]. Marine Pollution Bulletin, 2013, 75(1/2):126-132. Harrison J P, Schratzberger M, Sapp M, et al. Rapid bacterial colonization of low-density polyethylene microplastics in coastal sediment microcosms[J]. BMC Microbiology, 2014, 14:232. Amaral-Zettler L A, Zettler E R, Slikas B, et al. The biogeography of the plastisphere:implications for policy[J]. Frontiers in Ecology and the Environment, 2015, 13(10):541-546. De Tender C A, Devriese L I, Haegeman A, et al. Bacterial community profiling of plastic litter in the Belgian part of the North Sea[J]. Environmental Science & Technology, 2015, 49(16):9629-9638. Bryant J A, Clemente T M, Viviani D A, et al. Diversity and activity of communities inhabiting plastic debris in the North Pacific Gyre[J]. mSystems, 2016, 1(3):e00024-16. De Tender C A, Devriese L I, Haegeman A, et al. Temporal dynamics of bacterial and fungal colonization on plastic debris in the North Sea[J]. Environmental Science & Technology, 2017, 51(13):7350-7360. Oberbeckmann S, Osborn A M, Duhaime M B. Microbes on a bottle:substrate, season and geography influence community composition of microbes colonizing marine plastic debris[J]. PLoS One, 2016, 11(8):e0159289. Debeljak P, Pinto M, Proietti M, et al. Extracting DNA from ocean microplastics:a method comparison study[J]. Analytical Methods, 2017, 9(9):1521-1526. 周倩. 典型滨海潮滩及近海环境中微塑料污染特征与生态风险[D]. 烟台:中国科学院烟台海岸带研究所, 2016. Zhou Qian. Occurrences and ecological risks of microplastics in the typical coastal beaches and seas[D]. Yantai:Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, 2016. 章海波, 周倩, 周阳, 等. 重视海岸及海洋微塑料污染加强防治科技监管研究工作[J]. 中国科学院院刊, 2016, 31(10):1182-1189. Zhang Haibo, Zhou Qian, Zhou Yang, et al. Raising concern about microplastic pollution in coastal and marine environment and strengthening scientific researches on pollution prevention and management[J]. Bulletin of Chinese Academy of Sciences, 2016, 31(10):1182-1189. 阳静, 张静, 邹伟, 等. 环境微生物DNA提取方法研究进展[J]. 食品与机械, 2017, 33(3):207-210, 215. Yang Jing, Zhang Jing, Zou Wei, et al. Progress of study on extraction methods of environmental microbial DNA[J]. Food & Machinery, 2017, 33(3):207-210, 215. 吴敏娜, 武亚琦, 屈艳, 等. 四种小鼠肠道微生物DNA提取方法比较[J]. 生态学杂志, 2015, 34(4):1183-1188. Wu Minna, Wu Yaqi, Qu Yan, et al. Comparison of four methods for extracting microbial DNA from mouse intestine[J]. Chinese Journal of Ecology, 2015, 34(4):1183-1188. Murray J R, Rajeevan M S. Evaluation of DNA extraction from granulocytes discarded in the separation medium after isolation of peripheral blood mononuclear cells and plasma from whole blood[J]. BMC Research Notes, 2013, 6:440. Islam M R, Sultana T, Joe M M, et al. Comparisons of direct extraction methods of microbial DNA from different paddy soils[J]. Saudi Journal of Biological Sciences, 2012, 19(3):337-342. Donlan R M. Biofilms:microbial life on surfaces[J]. Emerging Infectious Diseases, 2002, 8(9):881-890. 熊志远. 聚苯乙烯泡沫(EPS)力学行为的实验研究[D]. 湘潭:湘潭大学, 2007. Xiong Zhiyuan. Experimental study on mechanical behaviors of expanded polystyrene(EPS)[D]. Xiangtan:Xiangtan University, 2007. Stasiek J. Modern technologies and equipment for blowing extrusion of the films. Part Ⅰ. Blowing extrusion of the films from plastics[J]. Polimery, 2005, 50(3):169-175. 陈优霞. 二氧化硅塑料薄膜开口剂的研制[D]. 南昌:南昌大学, 2011. Chen Youxia. Study on preparation of silica for plastic films of the anti-block agent[D]. Nanchang:Nanchang University, 2011.
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