Zhao Huabing, Shi Sanyuan, Zhao Hong, Guo Jin, Yang Zhen, Gao Hongsheng, Lu Fuping
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, National Engineering Laboratory for Industrial Enzymes, College of Biotechnology, Tianjin University of Science & Technology Tianjin 300457 China
Animals, Plants and Food Testing Center of Tianjin Customs District Tianjin 300461 China.
RSC Adv. 2019 Dec 16;9(71):41628-41638. doi: 10.1039/c9ra06004a. eCollection 2019 Dec 13.
Increasing attention has been paid to the toxicity and hazards of antibiotics on non-target organisms in soil ecosystems because redundant antibiotics in the excretion of treated animals are being brought into the soil by way of manure and sewage irrigation. In order to understand the toxic mechanisms of antibiotics in soil ecosystems, the earthworm was exposed to 500 mg kg of oxytetracycline (OTC) as a typical antibiotic for 7, 14 and 21 days. The total proteins of in each treatment were separated by two-dimensional gel electrophoresis and differential expressed proteins were identified by MALDI-TOF/TOF-MS. A total of 30 proteins were successfully identified and divided into four categories based on the function. It was surprisingly found that more than 50% of identified proteins belong to the actin family, and all of them were down-regulated more than 2.0-fold. In the meantime, the fibrinolytic enzymes, an important protease with plasminogen activator activity, were suppressed in the last two weeks. The validations in the mRNA level were performed using RT-PCR. However, due to the incomplete genome sequence of , we failed to identify more proteins response to OTC stress. This study may provide a new insight into the discovery of novel biomarkers for continuous-poured and low-toxicity pollutants.
由于经处理动物排泄物中的多余抗生素通过粪便和污水灌溉进入土壤，抗生素对土壤生态系统中非靶标生物的毒性和危害已受到越来越多的关注。为了了解抗生素在土壤生态系统中的毒性机制，将蚯蚓暴露于500 mg/kg的土霉素（OTC）（一种典型抗生素）中7天、14天和21天。通过二维凝胶电泳分离各处理中的总蛋白质，并通过基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF/TOF-MS）鉴定差异表达的蛋白质。共成功鉴定出30种蛋白质，并根据功能分为四类。令人惊讶的是，发现超过50%的已鉴定蛋白质属于肌动蛋白家族，并且所有这些蛋白质的表达下调均超过2.0倍。同时，在最后两周，具有纤溶酶原激活剂活性的重要蛋白酶——纤溶酶受到抑制。使用逆转录聚合酶链反应（RT-PCR）进行mRNA水平的验证。然而，由于蚯蚓基因组序列不完整，我们未能鉴定出更多对OTC胁迫有反应的蛋白质。本研究可能为持续倾倒和低毒性污染物新型生物标志物的发现提供新的见解。
