环境浓度氧四环素与聚苯乙烯微塑料对黄颡鱼幼鱼肠道的联合毒性效应
Combined Effects of Environmental Concentration of Oxytetracycline and Polystyrene Microplastics on Intestinal Tract of Juvenile Yellow Catfish (Pelteobagrus fulvidraco)
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摘要: 近年来，氧四环素(oxytetracycline, OTC)和聚苯乙烯微塑料(polystyrene microplastics, PS-MPs)在水环境中被广泛检出。为了探究PS-MPs与OTC对鱼类的联合毒性效应，选取黄颡鱼(Pelteobagrus fulvidraco)幼鱼为研究对象，将其暴露于CON(对照)组、环境浓度OTC(500 ng·L-1)单独组、MPs-L(100 μg·L-1 PS-MPs)单独组、MPs-L+OTC(100 μg·L-1 PS-MPs+500 ng·L-1 OTC)复合组、MPs-H(1 000 μg·L-1 PS-MPs)单独组和MPs-H+OTC(1 000 μg·L-1 PS-MPs+500 ng·L-1 OTC)复合组中28 d，研究了OTC和PS-MPs单独以及联合暴露对幼鱼生长、肠道结构和肠道菌群的影响。研究结果表明，与对照组相比，OTC单独暴露和MPs-L单独暴露对黄颡鱼幼鱼体长、体质量及体质量增长率，肠道氧化应激酶(超氧化物歧化酶(superoxide dismutase, SOD)和过氧化氢酶(catalase, CAT))活性及消化酶(胰蛋白酶(trypsin, TRS)、淀粉酶(amylase, AMS)和脂肪酶(lipase, LPS))活性，肠道微生物组成(OTU数目、α多样性、β多样性以及门、属水平上物种组成的相对丰度)均无显著性影响。但MPs-L+OTC复合暴露导致SOD和CAT活性显著升高，引起肠道空泡化、肠上皮细胞轻微缺失，变形菌门的相对丰度显著升高，且与OTC单独暴露组相比，肠道CAT活性显著性升高。MPs-H单独暴露抑制了黄颡鱼幼鱼体质量和体质量增长率，引起了肠道空泡化，导致其肠道SOD和CAT活性显著升高，消化酶TRS和LPS活性显著降低，厚壁菌门相对丰度显著降低。与MPs-H单独组和OTC单独组相比，MPs-H+OTC组进一步加剧肠道氧化酶活性升高、消化酶活性降低、肠道损伤和肠道菌群紊乱。相关性分析表明，肠道葡萄球菌属和体长显著负相关；鲸杆菌属和SOD显著正相关；气单胞菌属与LPS显著负相关，与AMS显著正相关。上述结果显示PS-MPs高浓度单独以及与OTC复合暴露可能通过肠道损伤以及肠道菌群的改变，进而影响肠道消化酶活性，导致黄颡鱼幼鱼生长抑制。此外，PS-MPs和OTC的复合肠道毒性表现出显著的协同效应。本实验结果将为水环境中抗生素和微塑料的生态风险评价提供新的视角和理论依据。Abstract: In recent years, oxytetracycline (OTC) and polystyrene microplastics (PS-MPs) have been widely detected in aquatic environment. In order to investigate the combined effects of PS-MPs and OTC on intestinal tract of fish, we investigated the effects of OTC and PS-MPs exposure on growth, intestinal structure and intestinal microflora of fish. Juvenile yellow catfish (Pelteobagrus fulvidraco) were exposed to 500 ng·L-1 OTC (OTC), 100 (low concentration) (MPs-L) and 1 000 μg·L-1 (high concentration) (MPs-H) PS-MPs, or their combination (combined MPs-L+OTC, combined MPs-H+OTC) for 28 d. Results showed that OTC and MPs-L alone exposure had no significant effect on the growth, intestinal antioxidant enzyme activities (including superoxide dismutase (SOD) and catalase (CAT)), digestive enzyme activities (including trypsin (TRS), amylase (AMS) and lipase (LPS)), or intestinal flora (including OTU number, alpha diversity, beta diversity, and relative abundance of species composition at phylum level and genus level) of juvenile yellow catfish. However, combined MPs-L+OTC exposure significantly increased SOD and CAT activities, induced intestinal vacuolation and slight loss of intestinal epithelial cells, as well as significantly increased the relative abundance of Proteobacteria as compared to the control group. Intestinal CAT activity was significantly increased in the combined MPs-L+OTC exposure group as compared to the OTC alone exposure group. Moreover, MPs-H alone exposure inhibited the body weight and weight gain rate of juvenile yellow catfish, induced intestinal vacuolation, significantly increased the activities of SOD and CAT, significantly decreased the activities of digestive enzymes TRS and LPS, and significantly decreased the relative abundance of Firmicutes. Compared with the MPs-H group and OTC group, the effects on oxidase activities, digestive enzyme activities, intestinal injury and intestinal flora were further exacerbated in the combined MPs-H+OTC group. In addition, correlation analysis showed that a significant negative correlation between Staphylococcus and body length. Abundance of Cetobacterium was positively correlated with the activities of SOD. Aeromonas was negatively correlated with the activity of LPS while positively correlated with AMS. Thus, high concentration of PS-MPs alone or combined OTC exposure might affect intestinal digestive enzyme activities through intestinal injury and changes in the intestinal flora, resulting in growth inhibition of juvenile yellow catfish. In addition, the combined intestinal toxicity of PS-MPs and OTC showed significant synergistic effects. The results of this study might provide a new perspective and theoretical basis for ecological risk assessment of antibiotics and microplastics in aquatic environment.
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