广州市兽用抗生素的环境残留研究

畜禽废物中的抗生素可造成土壤及水体的抗生素污染,利用高效液相色谱及高效液相色谱-串联质谱分析了广州市代表性养殖场畜禽废物、施用畜禽粪土壤和鱼塘水中尼卡巴嗪、喹乙醇、四环素类、磺胺类、喹诺酮类、氯霉素类抗生素的含量。结果发现：猪粪、鸡粪中抗生素含量最高的均为四环素类,分别为123.76、14.59 mg.kg-1;含量最低的均为氯霉素类,分别为2.35μg.kg-1、0.08 mg.kg-1;小猪猪粪中抗生素的含量明显高于大猪。鱼塘水中抗生素含量较高的是四环素（5.16μg.L-1）与磺胺对甲氧嘧啶（4.78μg.L-1）,土霉素在所有样点中均未被检出。施用禽畜粪土壤中四环素类、喹诺酮类含量较高,分别为70.40、49.77μg.kg-1;磺胺甲基嘧啶、磺胺对甲氧嘧啶、磺胺甲噁唑、甲砜霉素、氯霉素、喹乙醇均未被检出。粪样中的抗生素可导致土壤和水体的抗生素污染,TCs、QNs和NCZ可能对生态环境和人类健康安全的威胁更大。     

生物质炭对磺胺类抗生素在坡耕地紫色土中吸附-解吸及淋溶过程的影响

吸附–解吸是影响抗生素类污染物在土壤中迁移转化及生物有效性的重要过程,本文以川中丘陵区坡耕地紫色土为研究对象,通过批量平衡实验和柱实验研究生物质炭施用(投加量0(B0)、39.75 t/hm~2(B1)和198.75 t/hm~2(B2))及田间老化作用(夏季干湿交替)对3种典型磺胺类抗生素(磺胺嘧啶(SD)、磺胺二甲基嘧啶(SM2)及磺胺甲恶唑(SMZ))在紫色土中的吸附–解吸和淋溶行为的影响。结果表明:在几种处理中,3种磺胺类抗生素吸附强弱的顺序都表现为SDSMZSM2;与B0处理相比,添加生物质炭能增加土壤对3种抗生素的吸附能力,其中SM2的吸附显著增加(P0.05),但这种促进作用在经过老化过程后有所减弱。在解吸过程中,3种抗生素的Freundlich常数Kf大小顺序为SDSMZSM2,表明SD在土壤中吸附容量最大且不易解吸,其次是SMZ和SM2;相应的迟滞系数H大小顺序为SDSMZSM2,与Freundlich常数n值趋势一致,表明紫色土对SD亲和力最强,解吸最难,而SM2则解吸最易,可逆性最强。与B0处理土柱相比,3种磺胺抗生素在B1处理土柱中迁移更慢。     

生物质炭对磺胺类抗生素在坡耕地紫色土中吸附-解吸及淋溶过程的影响

吸附-解吸是影响抗生素类污染物在土壤中迁移转化及生物有效性的重要过程,本文以川中丘陵区坡耕地紫色土为研究对象,通过批量平衡实验和柱实验研究生物炭施用（投加量0(B0)、39.75 (B1)t/hm2和198.75 (B2) t/hm2）及田间老化作用（夏季干湿交替）对三种典型磺胺类抗生素（磺胺嘧啶（SD）、磺胺二甲基嘧啶（SM2）及磺胺甲恶唑（SMZ））在紫色土中的吸附-解吸和淋溶行为的影响。结果表明：在几种处理中,三种磺胺类抗生素吸附强弱的顺序都表现为SD>SMZ>SM2;与B0处理相比,添加生物炭能增加土壤对三种抗生素的吸附能力,其中SM2的吸附显著增加（P<0.05）,但这种促进作用在经过老化过程后有所减弱。在解吸过程中,三种抗生素的Freundlich常数Kf大小顺序为SD>SMZ>SM2,表明SD在土壤中吸附容量最大且不易解吸,其次是SMZ和SM2;相应的迟滞系数H大小顺序为SD相似文献   

土壤磺胺类抗生素固相萃取-高效液相色 谱法的构建与优化

设计正交试验对红壤中磺胺甲恶唑(sulfamethoxazole, SMZ)的萃取方法进行优化,并结合高效液相色谱,构建了固相萃取-高效液相色谱联合测定法(SPE-HPLC),并将该方法应用于不同抗生素以及不同类型土壤中SMZ的提取测定。结果表明,在以乙腈–磷酸盐缓冲液为提取液、提取液量为10 ml、超声时间为15 min、9 ml甲醇淋洗固相萃取柱(HLB)的条件下,红壤中SMZ的提取效果最优,0.25 mg/kg的SMZ回收率达到85.58%,能够满足环境样品的分析要求。利用该法提取测定红壤中不同抗生素包括3种磺胺类抗生素(磺胺嘧啶、磺胺二甲基嘧啶、磺胺甲基嘧啶)和2种四环素类抗生素(土霉素、盐酸四环素),结果表明,当抗生素浓度为0.25 mg/kg时,磺胺类抗生素(SAs)的回收率范围在67.31%～85.58%,四环素类抗生素(TCs)的回收率范围在20.81%～59.33%。利用该法分别提取测定不同类型土壤中的SMZ回收率,得到潮土中SMZ的回收率最大,达到92.92%,其次为荒漠土、红壤、紫色土,最低的回收率出现在黄棕壤,仅为53.62%。据相关性分析表明,回收率与土壤电导率(EC)、微生物碳氮比(C/N)呈极显著负相关(P0.01),与阳离子交换量(CEC)、有机质(SOM)呈显著负相关(P0.05)     

砂质潮土中五种磺胺类药物的高效液相色谱检测方法

建立了固相萃取-高效液相色谱法(SPE-HPLC)测定砂质潮土中磺胺嘧啶、磺胺甲基嘧啶、磺胺甲氧嗪、磺胺-5-甲氧嘧啶、磺胺甲恶唑5种磺胺类药物残留的方法。优化后的色谱条件为:流动相比例为磷酸缓冲液(pH=2.3)∶乙睛=80∶20(V/V),流速1.0 mL/min,检测波长270 nm,柱温35℃,进样体积20μL。在优化条件下,5种磺胺类药物的最低检测限在2.4~4.9μg/kg之间,方法的线性范围为0.2~20 mg/kg,线性相关系数均达到0.999 9。砂质潮土中,在添加浓度为0.5、1.5 mg/kg磺胺类药物时平均回收率范围为70.03%~89.65%,相对标准偏差≤10%,能够满足实际样品的分析要求。     

磺胺甲恶唑在沉积物中的降解行为研究

为评价磺胺类抗生素在沉积物中降解行为,以最常用的磺胺类药物——磺胺甲恶唑（SMZ）为对象,通过不同的环境条件下室内模拟实验,研究了其在沉积物中的降解动态以及相关环境因素（微生物、含氧量、光照、沉积物种类和药物起始浓度等）对降解过程的影响情况。结果表明,SMZ在沉积物中的降解途径主要为兼性厌氧微生物降解,非生物降解等其他降解途径只占较小比例。SMZ的降解速率与沉积物的有机质含量密切相关,高有机质含量的沉积物中SMZ的降解更快,沉积物中的光敏剂也能促进SMZ的降解。高浓度的SMZ通过抑制沉积物微生物的活性,使得其降解显著下降。     

施肥方式对蔬菜地土壤中8种抗生素残留的影响

从浙江省杭州、嘉兴和绍兴等3个地级市采集了4种不同施肥方式下(分别为施用畜禽粪+化肥、商品有机肥+化肥、沼渣+化肥和单施化肥)的蔬菜地表层土壤样品44个,分析了4类8种抗生素(包括四环素类抗生素的土霉素、四环素和金霉素,喹诺酮类抗生素的恩诺沙星,磺胺类抗生素的磺胺嘧啶、磺胺二甲嘧啶和磺胺甲噁唑及大环内脂类抗生素的泰乐菌素)的残留情况,探讨了施肥方式对蔬菜地土壤中抗生素残留的影响。结果表明,蔬菜地土壤中抗生素的检出率和残留含量与施肥方式密切相关。8种检测的抗生素中土霉素的检出率和残留含量明显高于其他种类的抗生素,土霉素的平均含量占8种抗生素总量平均值的67.03%。抗生素的检出率和平均含量由高至低依次为:土霉素磺胺二甲嘧啶恩诺沙星四环素磺胺甲噁唑、泰乐菌素金霉素磺胺嘧啶;四环素类抗生素磺胺类抗生素。土壤中各类抗生素的检出率及含量均为施用畜禽粪的蔬菜地施用商品有机肥的蔬菜地施用沼渣的蔬菜地单施化肥的蔬菜地,施用畜禽粪的蔬菜地土壤中抗生素残留量明显高于其他蔬菜地。试验结果表明,畜禽粪是蔬菜地土壤抗生素的主要来源,商品有机肥和沼渣的施用对蔬菜地土壤中抗生素的残留也有一定的贡献。     

磺胺甲嗯唑单克隆抗体的研制及免疫金标试纸检测方法的建立

用磺胺甲嗯唑人工免疫原（BSA-SMZ）免疫BALB／C小鼠（Mus muscalus）,细胞融合技术筛选抗磺胺甲嗯唑单克隆抗体（SMZ mAb）杂交瘤细胞,体内诱生腹水法制备SMZ mAb,并鉴定其免疫学特性;应用SMZ mAb研制SMZ残留快速检测金标试纸（SMZ-strip）,并测定其性能。结果表明,筛选出3株杂交瘤细胞,其中最好的3G6株间接ELISA效价细胞培养上清为1：8.1&#215;10^2,腹水为1：6.4&#215;10^5,亲和常数（K=a）为3.07&#215;10^9L／mol,对SMZ的半数抑制浓度（IC50）为12.4μg／L,与磺胺嘧啶的交叉反应率（CR％）为2.84％,与其它磺胺类药物无交叉反应;SMZ-strip目测检测限为6μg/L;其敏感性与竞争ELISA试剂盒相当,符合率为100％;SMZ-strip具有快速、敏感、特异、简便等特点,适合于SMZ残留快速检测的推广应用。     

天津近郊地区淡水养殖水体的表层水及沉积物中典型抗生素的残留分析

利用固相萃取-高效液相色谱-串联质谱联用技术（SPE-HPLC-MS/MS）分析了天津近郊养殖水体的表层水及沉积物中喹诺酮类、四环素类及磺胺类3类共18种抗生素的残留,同时考察3类抗生素在不同季节的变化分布特征。结果表明,在淡水养殖水体的表层水中环丙沙星、恩诺沙星和土霉素等检出率较高,浓度范围在10.5～26.8μg·L-1之间;相对地,在养殖水体的沉积物中磺胺甲基异噁唑、磺胺甲噻二唑、磺胺二甲氧嘧啶、诺氟沙星的检出率较高,浓度范围在1.5～30.1μg·kg-1之间。研究同时发现,冬季3类抗生素在表层水和沉积物中的检出率和检出浓度普遍高于夏季。说明在我国部分淡水养殖地区的养殖水体中已经有一定量的磺胺类和喹诺酮类抗生素的残留,并可能威胁到水生生物的安全,进一步对人体健康产生危害,因此应加强对淡水养殖环境中抗生素残留的监测与管理。     

土壤反硝化对磺胺嘧啶及抗性基因消减的影响

农田土壤中抗生素及抗性基因的复合污染已给生态环境安全和人体健康带来了全新隐患。针对厌氧条件下,反硝化作用过程对土壤抗生素乃至抗性基因消减影响的研究一直相对较少。因而,本研究采集牛粪堆积池塘周边底层农田土壤作为目标污染土壤,重点研究反硝化作用过程对土壤磺胺嘧啶及抗性基因消减动态的影响。结果表明:相较于原始污染土壤处理(T1),添加了NO_3~–-N的处理(T2)可以显著强化土壤和水相中反硝化速率,提升N_2O的产气速率,促进土壤中磺胺嘧啶浓度和抗性基因丰度的快速降低;同时发现土壤反硝化基因(nir K、nir S和nos Z)与磺胺类抗性基因(sul Ⅰ和sul Ⅱ)呈显著负相关(P0.05),说明当NO_3~–-N底物越充足,土壤反硝化细菌活性往往被激活,其反硝化功能基因表达就越活跃,土壤反硝化作用过程就越强烈,从而反馈作用促进磺胺嘧啶抗生素的厌氧消减,进而有助于sul系列抗性基因丰度的显著衰减;同时通过高通量测序技术及对反硝化细菌的分离筛选后,发现变形菌门(Proteobacteria)赖氨酸芽胞杆菌属(Lysinibacillus)的细菌是土壤厌氧反应前后的主导优势菌群,对于强化反硝化过程和促进磺胺嘧啶及sul抗性基因的消减发挥了潜在的积极作用。本研究结果可为探明土壤中抗生素的厌氧消减过程和缓解抗性基因的扩散传播提供新颖的认知基础。     

Environmental fate of two sulfonamide antimicrobial agents in soil

Veterinary antimicrobial agents have been detected in a number of environmental samples, including agricultural soils. In this study, we investigated the persistence and sorption of the sulfonamides sulfamethazine (SMZ) and sulfachloropyridine (SCP) in soil and their potential effects on soil microorganisms. The sulfonamides dissipated more rapidly from the silt loam soil as compared to the sandy soil. Average half-lives of SMZ and SPC among the two soils were 18.6 and 21.3 days, respectively. The presence of liquid swine slurry (5% v/w) decreased sulfonamide persistence in the silt loam soil. The lower persistence of the antimicrobials in liquid swine slurry-amended soil was likely due to higher microbial activity, as compared to unamended soil, and/or to the greater bioavailability of the sulfonamides to degrading microorganisms, as estimated by sorption isotherms. Concentrations of SMZ and SPC up to 100 microg g-1 had no effect on antimicrobial degradation rates and soil microorganisms. These studies suggest that higher sulfonamide concentrations would be necessary to affect the main processes controlling their environmental fates in soil, but at the concentrations normally found in the environment, there would be little or no effects.     

绿色和有机蔬菜基地土壤中喹诺酮类抗生素的污染特征

利用高效液相色谱-串联质谱（HPLC-MS/MS）分析方法,探讨了广州市某绿色蔬菜基地和有机蔬菜基地土壤中4种喹诺酮类抗生素的含量与分布特征。结果表明,土壤中各化合物的检出率除洛美沙星为85%以外,其余均为100%,平均含量为0.80～24.95μg.kg-1,以诺氟沙星（24.95μg.kg-1）为主,其次为环丙沙星（15.40μg.kg-1）和恩诺沙星（7.68μg.kg-1）。4种喹诺酮类化合物总含量在7.15～122.25μg.kg-1之间,主要分布在50～100μg.kg-1之间,平均为48.85μg.kg-1。有机蔬菜基地土壤中4种喹诺酮类化合物的含量均高于绿色蔬菜基地土壤。同一基地不同蔬菜土壤中喹诺酮类抗生素的含量有一定差异,但化合物组成特征差异不大。研究区土壤中喹诺酮类抗生素的含量低于兽药国际协调委员会（VICH）筹划指导委员会的抗生素生态毒性效应触发值（100μg.kg-1）,表明生态风险较小。     

Combined effects of compost containing Sulfamethazine and zinc on pakchoi (Brassica chinensis L.) growth,soil sulfonamide resistance genes,and microbial communities

To understand the ecotoxicity of antibiotics and heavy metals in soil, 5% (w/w) composts containing different concentrations of sulfamethazine (SMZ) and/or zinc (Zn) were applied to soil to investigate their effects on pakchoi (Brassica chinensis L.) growth, soil sulfonamide resistance genes (SRGs: sul1, sul2, and dfrA7), and soil microbial communities. Composts containing less than 1.0 mg SMZ kg^–1 or less than 2.8 g Zn kg^–1 promoted pakchoi growth and the metabolic activity of soil microbial communities. Compared with the control, the absolute abundances (AAs) of soil SRGs significantly increased by 0.85–4.54 times with 50.6 μg kg^–1 SMZ treatment (P < 0.05), the AA of sul2 increased by 166% with 248.8 mg Zn kg^–1 treatment. The combination treatments with 19.8 μg SMZ kg^–1 and 179.9 mg Zn kg^–1 in soil had synergistic stimulatory effects on pakchoi growth, soil SRGs and microbial metabolism, whereas 53.5 μg SMZ kg^–1 and 262.1 mg Zn kg^–1 had an opposite effect. Pearson’s correlation analysis showed that carbon metabolism by soil microorganisms had significant positive correlations with shoot height of pakchoi (r = 0.84, P < 0.05) and AAs of SRGs (r > 0.80, P < 0.05).     

Investigation of sulfonamide, tetracycline, and quinolone antibiotics in vegetable farmland soil in the Pearl River Delta area, southern China

Thirteen antibiotics in soil from vegetable farmlands of the Pearl River Delta, southern China, were investigated. At least three antibiotics were detected in each sample. Six antibiotics including four quinolones, tetracycline, and sulfamethoxazole were detected in >94% of the samples. The total contents of three tetracyclines, eight sulfonamides, and four quinolones were not detected-242.6, 33.3-321.4, and 27.8-1537.4 μg/kg, respectively. The highest antibiotic concentrations were observed mainly in vegetable farmlands affiliated with livestock farms. Chlortetracycline, sulfameter, and quinolones in some samples exceed the ecotoxic effect trigger value (100 μg/kg) set by the Steering Committee of Veterinary International Committee on Harmonization. The composition and concentration of antibiotics in soil were correlated with vegetable species. This study has revealed an alarming condition of antibiotics in vegetable farmland soil. Further investigation including environmental fate, plant uptake, and human exposure to antibiotics by plant-derived food should be conducted.     

黑龙江农田土壤壬基酚及其短链聚氧乙烯醚残留调查

对黑龙江土壤样品中的壬基酚(NP)、壬基酚一氧乙烯醚(NP1EO)和壬基酚二氧乙烯醚(NP2EO)的残留状况开展研究。结果表明,NP、NP1EO和NP2EO均有不同程度的检出,检出率在68.46%~95.38%之间;NP、NP1EO和NP2EO的浓度分别为ND~156.8μg kg-1、ND~15.70μg kg-1和ND~13.63μg kg-1,均值分别为22.02μg kg-1、4.04μg kg-1和2.15μg kg-1;进一步分析,已检出的样品中,NP在0~10μg kg-1的比重最大,NP1EO在4~6μg kg-1比重最大,NP2EO在1~2μg kg-1比重最大。NP的残留水平高于NP1EO和NP2EO,说明NP相对较难降解,更易在土壤中累积。     

Sorption and transport of sulfonamides in soils amended with wheat straw-derived biochar: effects of water pH,coexistence copper ion,and dissolved organic matter

Purpose

Sulfonamides are widely used for the prevention and treatment of bacterial infections, hard-degraded contaminants distributed in the environment if they are discharged into the soil and water. Biochar could probably influence the geochemical behavior of ionized antibiotics in the soils.

Materials and methods

To determine the sorption/desorption of three representative sulfonamides (SAs) in soils amended with biochar, we investigated the effects of water pH, Cu²⁺, and dissolved humic acid on the sorption of sulfamethoxazole (SMX), sulfamethazine (SMZ), and sulfadiazine (SD) onto two different soil samples (S1 pH?=?5.13 and S2 pH?=?7.33) amended with wheat straw-derived biochar (size 0.5～0.6 mm).

Results and discussion

Batch experiments showed that the sorption/desorption isotherms of SAs on soil with/without biochar followed the Freundlich model. The biochar had a strong adsorption potential for SMX, SMZ, and SD both in S1 and S2 at low water pH. Except for SMX, the presence of Cu²⁺ inhibited the sorption of SMZ and SD through competing hydrophobic adsorption region in soils. HA suppressed the sorption of three sulfonamides in soil S2 by electrostatic repulsion under alkaline condition. The soil leaching column experiments showed the SA transport in soils, and S1 and S2 amended with biochar (0.5 and 1.0 wt%) brought about 12–20 % increase in SMX, SMZ, and SD retention compared to the untreated soil.

Conclusions

The results indicated that the presence of biochar effectively mitigated the mobility of ionized antibiotics such as SMX, SMZ, and SD in soils, which helps us reconsider the potential risk of antibiotics in the environment.

     

Development of an enzyme-linked immunosorbent assay for seven sulfonamide residues and investigation of matrix effects from different food samples

Direct competitive enzyme-linked immunosorbent assays (ELISA) were developed to detect a broad range of sulfonamides in various matrices. Screening for this class of antibiotics in pig muscle, chicken muscle, fish, and egg extracts was accomplished by simple, rapid extraction methods carried out with only phosphate-buffered saline (PBS) buffer. Twenty milliliters of extract solution was added to 4 g of sample to extract the sulfonamide residues, and sample extracts diluted with assay buffer were directly analyzed by ELISA; matrix effects could be avoided with 1:5 dilution of pig muscle, chicken muscle, and egg extracts with PBS and 1:5 dilution of fish extract with 1% bovine serum albumin (BSA)-PBS. For liver sample, the extraction method was a little more complicated; 2 g of sample was added to 20 mL of ethanol, mixed, and then centrifuged. The solvent of 10 mL of the upper liquid was removed, and the residues were dissolved in 10 mL of PBS and then filtered; the filtrate was diluted two-fold with 0.5% BSA-PBS for ELISA. These common methods were able to detect seven sulfonamide residues such as sulfisozole, sulfathiazole, sufameter, sulfamethoxypyridazine, sulfapyridine, sulfamethizole, and sulfachlorpyridazine in pig muscle, liver, chicken muscle, egg, and fish. The assay's detection limits for these compounds were less than 100 microg kg-1. Various extraction methods were tested, and the average recovery (n=3) of 100 microg kg-1 for the matrices was found to range from 77.3 to 123.7%.