蚯蚓对金霉素污染土壤酶活性和微生物群落的影响

为探究蚯蚓对抗生素污染土壤酶活性和微生物群落的影响,选取金霉素为目标污染物,威廉环毛蚓（内层种）和赤子爱胜蚓（表层种）为实验生物,通过高效液相色谱-串联质谱法、土壤酶试剂盒（微板法）、高通量测序技术和冗余分析对土壤中金霉素残留、酶活性和微生物群落进行了测定与分析。结果表明：经过28 d的培养,蚯蚓处理组中金霉素浓度（0.022 4~1.006 0 mg·kg^-1）显著低于对照组土壤（0.033 5~1.585 0 mg·kg^-1）。金霉素抑制了土壤脱氢酶、蔗糖酶、脲酶和碱性磷酸酶的活性,激活了过氧化氢酶的活性。两种蚯蚓均激活了土壤脱氢酶、过氧化氢酶、蔗糖酶和脲酶的活性,最大激活率分别为77.80%~88.50%、4.00%~6.16%、69.20%~72.60%、43.20%~48.80%;碱性磷酸酶的活性受到抑制,最大抑制率为23.8%~25.0%。高通量测序结果表明两种生态型蚯蚓均可改变土壤中细菌群落结构,不同蚯蚓类型对细菌群落结构的影响存在有差异。两种蚯蚓的加入均未改变土壤中优势菌门的组成,但都改变了其丰度占比。在属水平上,Flavobacterium、Aeromonas、Luteolibacter、Adhaeribacter和Pseudomonas等潜在金霉素降解菌被两种蚯蚓刺激丰度显著增加,加速了土壤中金霉素的降解。冗余分析结果表明,土壤中pH、有机质含量、金霉素浓度和细菌群落结构是影响土壤酶活性变化的重要因子。研究表明,威廉环毛蚓对脱氢酶、蔗糖酶和脲酶的促进作用优于赤子爱胜蚓。     

The fate of chlortetracycline residues in a simulated chicken–fish integrated farming systems

Two experiments were conducted at the Asian Institute of Technology, Pathumthani, Thailand to investigate the fate of chlortetracycline (CTC) residue in chicken manure and its effect on integrated chicken–fish farming system. During the first experiment, broiler chickens were raised and CTC residues in their manure were analysed. Chicken fed diets containing 0, 50, 200 and 800 CTC mg kg^?1 had CTC residue levels of 0, 0.9, 3.8 and 6.5 CTC ng g^?1. Once the diet containing CTC was withdrawn, CTC in the manure dropped to negligible amounts (0, 0, 0.2 and 0.5 CTC ng g^?1) within 1 day. Integrated chicken–fish farming systems were simulated during the second experiment to determine the fate of antibiotic residues in chicken manure in aquaculture environment. Chickens were fed a CTC‐free diet and a feed containing CTC at 200 mg kg^?1. Ten 4 m³ square concrete tanks (2 × 2 × 1 m) were used for the experiment. Five tanks were fertilized with CTC‐contaminated manure and the remaining five tanks were fertilized with CTC‐free manure at a rate of 100 kg dry matter ha^?1 day^?1. Sex‐reversed Nile tilapia (Oreochromis niloticus) was stocked at 12 fish tank^?1 on the 14th day after chicken manure application. The immuno‐radio microbial receptor assay (Charm II test) revealed that edible fish muscle, fish intestinal tract and sediment were contaminated by CTC at rates of 7.21, 22.104 and 1.788 ng g^?1, respectively, after 45 days. Chlortetracycline was detected on day 20 in the water column and gradually increased from 0.26 to 12.13 ng g^?1. Chlortetracycline residues were not detected in fish or the aquatic environment of the CTC‐free treatment. The results demonstrate the potential for antibiotic residue accumulation in fish and aquatic environment when CTC‐contaminated chicken manure is used for pond fertilization.     

金霉素和代谢物差向金霉素在肉鸡肌肉和肝脏中的残留分布

为了研究金霉素(CTC)、差向金霉素(ECTC)在肉鸡肌肉、肝脏中的残留分布情况,样品经提取液提取后过Oasis HLB SPE小柱净化,采用电喷雾正离子模式(ESI+)和多反应监测模式(MRM),用高效液相色谱-串联质谱仪检测;试验组鸡按体重以50 mg/(kg·d)剂量内服CTC,连续5 d,每天给药1次。结果表明:该方法测定肉鸡肌肉和肝脏中CTC检测限和定量限分别为5.0、10.0μg/kg。ECTC检测限和定量限分别为20.0、30.0μg/kg。CTC和ECTC在肉鸡肌肉中平均回收率分别在73.0%~95.20%和81.80%~101.60%,相对标准偏差(RSD)分别在8.85%~12.18%和2.05%~9.29%;在肉鸡肝脏中平均回收率分别在67.06%~90.80%和78.64%~106.98%,RSD分别在2.90%~8.88%和4.72%~9.48%;肉鸡肌肉和肝脏中CTC、代谢物ECTC的残留量分别在给药后第1、5 d时达到最高。肉鸡肝脏中ECTC的残留量约占总残留量的1/2。停药后CTC及其代谢物ECTC代谢缓慢,在停药第6 d时,仍能检测到CTC及其代谢物ECTC,但其残留量均低于最高残留限量(MRLs)。从保障食品安全方面考虑,建议我国将CTC和ECTC均作为金霉素的残留标示物进行监测。     

金霉素及其异构体降解产物对斜生栅藻的毒性效应研究

为探索水体中四环素类抗生素异构体降解产物的毒性效应,选取金霉素及其异构体降解产物为目标化合物,斜生栅藻(Scenedesmus obliquus)为受试生物,结合藻类生理指标探讨金霉素及其异构体降解产物的毒性效应。研究结果表明,在斜生栅藻藻液中,金霉素主要的异构体降解产物为异金霉素及异差向金霉素。暴露72 h后,金霉素及其异构体降解产物处理组的藻细胞形态发生了明显的质壁分离,且细胞通透性均显著增大。但是,金霉素母体药物与不同异构体降解产物对斜生栅藻的毒性效应表现出了明显的差异。金霉素母体药物处理组的斜生栅藻细胞中可溶性蛋白质和叶绿素a浓度降低得更为显著,且氧化损伤更为严重。金霉素母体药物与其异构体降解产物虽然具有相似的化学结构,但取代基空间构象的不同会导致药物与藻细胞中的可溶性蛋白质的结合位点不同,因而对藻细胞产生不同的毒性效应。探究金霉素及其异构体降解产物对水生环境产生的毒性效应,有望为全面认识四环素类抗生素的生态环境风险提供新的依据。     

4种典型抗生素对土壤微生物呼吸的影响

本研究通过室内直接吸收法测定了四环素、金霉素、诺氟沙星和恩诺沙星4种典型抗生素对土壤微生物呼吸作用的影响。结果表明,4种抗生素对土壤微生物呼吸的影响存在一定差异。四环素在整个处理过程中对土壤呼吸的影响以抑制作用为主;金霉素在处理第1d和7d均有激活作用,而且激活率最高可达到51.23%,但是在其他处理时间以抑制作用为主;诺氟沙星在整个处理过程中,浓度为0.2mg·kg-1和1mg·kg-1对土壤微生物呼吸都有一定的抑制作用,浓度为5、25、50mg·kg-1处理均为激活作用,激活作用随着处理浓度的增加而升高。恩诺沙星在整个处理过程中,所有处理对呼吸的影响变化趋势为先激活后抑制。总体来说,到处理第15d,所有抗生素处理的微生物呼吸作用基本恢复到对照水平,说明在本试验浓度处理下,微生物对这4种抗生素能够产生一定的适应性。     

利用CTC染色进行猪精子不同体外获能方法的比较

为了探讨不同方法对猪精子体外获能的影响,本试验以猪的新鲜精液为研究对象,分别经肝素上游法、钙离子载体诱导法和咖啡因诱导法进行精子不同获能时间的处理,并利用金霉素荧光染色法（chlortetracycline,CTC）对其获能效果进行检验。结果表明,采用肝素上游法孵育40 min处理后,精子的获能比例显著高于其他各处理组（P<0.05）,说明肝素上游法可以较好地诱发猪精子在体外获能。     

金霉素对肉鸡生长、肠道菌群和细菌酶、肠组织形态的影响

取1日龄AA商品代混合雏鸡160只,随机均分2组,分别饲喂添加含0 mg/kg(对照)和50mg/kg金霉素的玉米豆粕型日粮42 d.24 h光照,笼养,粉料饲喂,自由采食和饮水.结果表明:金霉素有明显的促生长作用,并显著改善1～21日龄段的饲料转化效率;金霉素可降低肠道梭菌和大肠杆菌数,但对总需氧菌、总厌氧菌和乳酸杆菌数均未产生显著影响;金霉素能降低肠道中β-葡萄糖苷酶和β-葡萄糖苷酸酶的活性,改善小肠粘膜组织的形态结构.     

高效液相色谱法测定金霉素药渣中金霉素的含量

采用反相高效液相色谱法测定金霉素药渣中金霉素含量的方法研究。用C18分析柱 ( 3 .9× 15 0mm ,5 μm)。 0 .1mol/L草酸铵 -二甲基甲酰胺 - 0 .2mol/L磷酸二氢钾 ( 68:2 7:5 )为流动相。紫外检测器 :λ =3 70nm ,柱温 40℃ ,在 2 .5 - 10 0 μg/mL之间呈良好的线性关系 ,相关系数R =0 .9991,平均回收率为 98.3 4%。本法简便、快速、准确。适合于金霉素药渣及饲料中金霉素含量的测定     

水溶性有机质对金霉素在猪粪中吸附-解吸附行为的影响

选择来自猪粪和稻草的水溶性有机质（DOM）,用室内模拟试验方法研究了我国常用抗生素金霉素（CTC）在受到内源和外源DOM作用下在猪粪中的吸附和解吸附行为。研究表明,金霉素能被猪粪便快速吸附,等温线呈非线性,用Freundlich方程可以很好地拟合吸附数据;添加DOM能抑制猪粪对CTC的吸附,其能力大小与加入的DOM种类有关,其中内源性猪粪DOM的解吸附效果明显大于外源性稻草DOM;形成阳离子复合体和加入DOM的性质可能是抑制CTC在猪粪便上吸附的主要机制。     

Effects of straw addition on increased greenhouse vegetable yield and reduced antibiotic residue in fluvo-aquic soil

Organic manure application is an important measure for high yield and good quality vegetable production, whereas organic manure is also a main source of residual antibiotic in soils. A 3-yr experiment was conducted on a fluvo-aguic soil in Tianjin of northern China. The objective of this study was to investigate the effects of different fertilization patterns on yield of six-season vegetables with celery and tomato rotation, and dynamic change of tetracyclines residues in the soil during the sixth growing season(tomato season). The field experiment comprised six treatments depending on the proportion of nitrogen of each type of fertilizer: 4/4 CN(CN, nitrogen in chemical fertilizer), 3/4 CN+1/4 MN(MN, nitrogen in pig manure), 2/4 CN+2/4 MN, 1/4 CN+3/4 MN, 2/4 CN+1/4 MN+1/4 SN(SN, nitrogen in corn straw), and CF(conventional fertilization, the amounts of nitrogen application were 943 and 912 kg N ha–1 for celery and tomato season, respectively). In addition to CF treatment, the amount of nitrogen application in other treatments was greatly reduced and equal(450 and 450 kg N ha–1 for celery and tomato season, respectively). Results showed that the combined application of 3/4 CN+1/4 MN achieved the highest yield and economic benefit in the first four seasons, but addition of straw(2/4 CN+1/4 MN+1/4 SN treatment) performed better in the subsequent two seasons, and the average yields of 2/4 CN+1/4 MN+1/4 SN treatment were respectively higher by 9.9 and 12.8% than those of 4/4 CN treatment, and by 5.6 and 10.5% than those of CF treatment. The residual chlortetracycline(CTC) in manure-amended soil for three consecutive years increased along with the increase of applied amount of pig manure. Under the same amount of pig manure application, content of CTC in straw-amended soil was obviously decreased compared with no straw-amended soil(3/4 CN+1/4 MN treatment), and averagely decreased by 41.9% for four sampling periods in the sixth season. Addition of crop straw facilitated the degradation of CTC in manure-amended soil. As a whole, the conventional fertilization was not the desirable pattern based on yield, economic benefit and environment, the optimal fertilization pattern with the highest yield and profit and the least soil chlortetracycline residue was the treatment of2/4 CN+1/4 MN+1/4 SN under this experimental condition.