摘要
在海水养殖过程中,磺胺甲恶唑(sulfamethoxazole, SMX)等抗生素类药物的大量残留加速了抗性细菌和抗性基因的传播,严重威胁生态环境健康。生物法控制抗生素废水是解决其环境危害的重要途径。
目的
从近海养殖池底泥中筛选出一株耐盐且对SMX具有高效降解能力的菌株LS-1,分析环境因素对其降解能力的影响,优化菌株对SMX的降解性能,并通过产物类型解析其降解途径,最终对降解产物进行毒性分析。
方法
通过对分离菌株进行16S rRNA基因序列测序与系统发育树分析进行鉴定,采用单因素和响应面试验对降解条件进行优化,利用气质色谱法及发光细菌水质急性毒性试验检测分析其降解产物及产物毒性。
结果
分离获得的菌株LS-1与产碱杆菌属(Alcaligenes)中的水生产碱菌(Alcaligenes aquatilis) AS1序列相似度达99.79%。单因素试验确定胰蛋白胨是菌株生长和降解SMX时的最佳外源碳源。菌株在温度20-35 ℃、盐度15‰-35‰、SMX浓度10-100 mg/L、pH 7.0-9.0的条件下时生长良好。响应面分析表明,对SMX降解率有显著影响的因素依次为:SMX浓度>初始pH>环境温度。在SMX浓度为33 mg/L、pH 7.4和30 ℃的条件下,该菌株在48 h内的最高降解率达60.17%。质谱检测分析推测菌株LS-1通过乙酰化和羟基化等途径降解SMX,发光细菌急性毒性试验表明在SMX降解过程中生物毒性逐渐降低。
结论
本研究分离的SMX降解菌能够很好地适应海洋环境条件,降低SMX的水质毒性,对海水养殖废水中抗生素污染的防治具有重要的应用前景。
近年来,受全球,特别是亚洲水产养殖业增长的推动,全球渔业和水产养殖产量创历史新
针对SMX污染的控制,通常采用物理吸附以及化学氧化等技术,如生物炭、碳纳米管、芬顿反应以及光化学催化
响应面法是一种通过建立数学模型来分析多个显著影响因素与响应值之间的关系,并通过统计学方法验证模型结果的技术,已被广泛应用于多种污染物降解条件的优化,以提高去除效率和降低成
1 材料与方法
1.1 实验样品及试剂
海水养殖底泥与海水样品取自山东省青岛市即墨区鳌山湾森林公园附近的海水养殖池(120°69′E,36°37′N),养殖池水深0.2 m。取池底泥水混合物放入无菌样品瓶中,24 h内运回实验室,4 ℃冷藏保存。
磺胺甲恶唑(C10H11N3O3S)购自上海麦克林生化科技股份有限公司;色谱级乙腈、磷酸(98%)购自上海阿拉丁生化科技股份有限公司。实验使用的其他药品均为分析纯。
1.2 培养基
富集培养基(g/L):NH4Cl 1.0,K2HPO4 1.5,KH2PO4 0.5,MgSO4·7H2O 0.05,CH3COONa 0.2,NaCl 30.0,微量元素0.1 mL/L。微量元素(g/L):MgSO4·7H2O 0.5,EDTA 1.0,ZnSO4 0.2,MnCl2·4H2O 0.1,FeSO4·7H2O 0.5,CuSO4·5H2O 0.5,CoCl2·6H2O 0.2。降解培养基(g/L):K2HPO4 1.5,KH2PO4 0.5,MgSO4·7H2O 0.05,NaCl 30.0,Tryptone 2.0。LB液体培养基(g/L):胰蛋白胨 10,酵母浸粉 5,NaCl 30。LB固体培养基在上述成分基础上加入12 g/L琼脂。所有培养基经121 ℃灭菌20 min。
SMX母液:以无水乙醇为溶剂,配制SMX质量浓度为10 mg/mL,置于棕色瓶中,于‒20 ℃保存。人工海水配制:以蒸馏水为溶剂,NaCl浓度为30 g/L。
1.3 菌株的富集与纯化
取养殖池泥水样品5 g置于250 mL无菌海水中,30 ℃、150 r/min振荡3 h,使泥中细菌充分分散到无菌海水中。取5 mL混合样品转移至装有95 mL富集培养基的锥形瓶中,设置SMX初始浓度为5 mg/L,在恒温培养箱中30 ℃、120 r/min避光培养。7 d后转接至10 mg/L SMX的新鲜富集培养基中,依次重复上述步骤,每次转接按5 mg/L梯度浓度增大培养基中SMX含量,直至抗生素浓度达到30 mg/L。将富集菌群按1
1.4 菌株的分析鉴定
将筛选出的单菌株采用平板划线法接种于含有SMX的富集琼脂培养基上,30 ℃培养48 h后观察菌落形态并进行革兰氏染色。将菌株在LB培养基中30 ℃、120 r/min培养30 h至OD600值为0.5左右,20 ℃、5 000 r/min离心5 min收集菌体,用戊二醛固定过夜,经过乙醇脱水,冷冻干燥,表面喷金后,通过扫描电镜(Thermo Fisher 公司)观察菌体形态。将培养至对数生长期的菌株,取2 mL置于离心管中,委托青岛派森诺生物科技有限公司进行16S rRNA基因序列分析鉴定。得到的序列结果提交至NCBI数据库,通过BLAST进行初步对比鉴定。采用MEGA 11.0软件中的邻接(neighbor-joining)法进行多序列比对,构建系统发育树并分析结果。
将培养至对数生长期OD600为0.5左右的菌株,以2%的接种量接种至100 mL降解培养基中,于30 ℃、120 r/min振荡培养,分别在4-72 h使用紫外分光光度计测量菌株OD600。以时间为横坐标,OD600为纵坐标绘制菌株生长曲线。
1.5 菌株对SMX降解性能优化
改变不同影响因素,如碳源(胰蛋白胨、醋酸钠、葡萄糖、蔗糖)、SMX初始浓度(10、20、50、100 mg/L)、接种量(1%、3%、5%、7%)、盐度(20‰、25‰、30‰、35‰)、温度(20、25、30、35 ℃)、pH (5.0、7.0、9.0),采用控制单一变量的方法,探究各因素对菌株48 h降解率的影响。上述实验在120 r/min恒温振荡培养,每隔12 h取样,通过高效液相色谱(HPLC)分析培养液中SMX的剩余量。
通过Design-Expert 13设计Plackett-Burman(PB)实验,筛选对响应值具有显著影响的因素,用于后续的响应面分析(Box-Behnken方法)。设计响应面试验,分析影响因素的交互作用,优化降解条件。
1.6 降解产物检测及途径推断
利用安捷伦气相质谱仪(Agilent公司),对样本中的降解产物进行质谱分析。首先将样本在70 ℃下平衡4 min,随后以25 ℃/min的升温速率加热至100 ℃,并在此温度下保持1 min;接着,以3 ℃/min的升温速率继续升温至260 ℃,并在该温度下维持10 min,以确保充分分离与检测。质谱仪采用电子碰撞电离模式,电离能量设置为70 eV,溶剂延迟时间为4 min。MS源温度设定为230 ℃,传输线温度设定为280 ℃。选择全扫描模式,扫描范围为50-550 m/z。依据分析得出的降解产物,推测降解路径。
1.7 降解产物的毒性评估
为检测降解产物的毒性,采用明亮发光杆菌(Photobacterium phosphoreum) T3菌株,利用LumiFOX2000型生物毒性分析仪(深圳朗石公司)检测相对发光率的变化,以表征水质毒性。首先用2% NaCl复苏发光细菌冻干粉,复苏2 min后细菌在黑暗中发出明显荧光。在测试仪配备的试管中加入2% NaCl和10 μL复苏的发光细菌菌液,混匀后先测试无降解产物对照组的发光亮度,再依次测试含有降解产物的处理组发光亮度。相对发光率的计算如
相对发光率(%)=×100 | (1) |
1.8 分析方法
使用HPLC检测SMX含量。将经过微生物降解的培养液在4 ℃、4 000×g离心5 min,收集上清液,采用孔径为0.22 μm的针头过滤器过滤后,注入棕色液相瓶,4 ℃冷藏保存。使用高效液相色谱仪(1260 II,Agilent公司),配备C18反向色谱柱(4.6 mm×250 mm,5 μm,Agilent公司),流动相为色谱级乙腈和0.05 mol/L磷酸(体积比30:70),设置UV检测器波长为256 nm,柱温30 ℃,进样量10 μL,流量1.0 mL/min。
每个实验均重复3次,数据结果以平均值±标准差表示,CK表示空白对照。采用SPSS 16.0进行单因素方差分析及Tukey’s HSD检验,将P值阈值设置为小于0.05,以确定组间统计学上的显著差异。借助Origin 2021软件进行数据可视化。
2 结果与讨论
2.1 降解菌株的分离及鉴定
从养殖池采集的沉积物样品中,经过不断富集驯化,分离出5株具有对SMX耐药性的菌株,分别命名为LS-1、LS-2、LS-3、LS-4和LS-5。48 h内,各菌株对SMX的降解率分别为:LS-1为48%,LS-2为14%,LS-3为12%,LS-4为22%,而LS-5基本无降解能力。根据菌株对磺胺甲恶唑的降解率,选择LS-1作为实验菌株进行后续研究。
LS-1在平板上经30 ℃恒温培养48 h后,长出肉眼可观察的菌落,如

图1 菌株LS-1形态特征
Figure 1 Morphological characteristics of strain LS-1. A: Plate scribing; B: SEM image.
将菌株的16S rRNA基因序列上传至NCBI数据库进行同源性分析,结果表明,LS-1的16S rRNA基因序列与水生产碱菌(Alcaligenes aquatilis) AS1有99.79%的一致性。使用MEGA 11.0软件,通过邻接法构建系统发育树,如

图2 基于16S rRNA基因序列构建的菌株LS-1系统发育树
Figure 2 Phylogenetic tree of strain LS-1 based on 16S rRNA gene sequence. Red font: Labeled the strain screened; GenBank accession numbers are given in parentheses; Bootstrap values were expressed as a percentage of 1 000 replications; Bar 0.02 substitutions per nucleotide position.
2.2 菌株生长曲线
菌株LS-1的生长曲线(OD600)以及培养基中SMX的去除率如

图3 LS-1生长曲线及20 mg/L SMX降解率变化
Figure 3 LS-1 growth curve and changes in degradation efficiency of 20 mg/L SMX.
从SMX的降解曲线来看,在0 h到约10 h之间,SMX降解率出现短暂下降,可能是由于培养基pH变化导致SMX吸光度发生改
2.3 环境因子对降解菌株降解能力的影响
2.3.1 碳源对菌株LS-1降解磺胺甲恶唑的影响
菌株LS-1在以SMX为唯一碳源的培养基中无法生长,推测其对SMX的降解为共代谢作用。因此,设置实验探究LS-1在不同碳源(胰蛋白胨、醋酸钠、葡萄糖、蔗糖)条件下的SMX降解率及生长情况,以确定菌株生长和降解的最佳碳源。如

图4 不同影响因素条件下菌株LS-1对SMX的降解
Figure 4 Degradation of SMX by strain LS-1 under different influencing factors. A: Different carbon sources. B: Different salinities. C: Different pollutant concentrations. D: Ambient temperature. E: Different inoculum sizes. F: Different pH. The bar chart shows the degradation efficiency within 48 h.
2.3.2 盐度对菌株LS-1降解磺胺甲恶唑的影响
研究了不同盐度(15‰、25‰、30‰、35‰)条件下菌株LS-1对SMX降解的影响。结果表明,菌株在不同盐度条件下表现出较强的适应能力(
2.3.3 SMX浓度对菌株LS-1降解能力的影响
初始SMX浓度对菌株LS-1的降解能力有显著影响。如
2.3.4 环境温度对菌株LS-1降解磺胺甲恶唑的影响
温度是影响微生物生长和代谢的重要因素,通过影响细胞内酶的活性、细胞质的流动性以及污染物的性质来调节微生物的生长速
2.3.5 接种量对菌株LS-1降解磺胺甲恶唑的影响
从动力学角度来看,不同接种量条件下菌株LS-1对SMX的降解趋势基本一致(
2.3.6 初始pH对菌株LS-1降解磺胺甲恶唑的影响
环境pH对微生物的生长代谢、表面吸附作用以及抗生素的水解等过程具有重要影响。如
2.4 响应面优化及分析
2.4.1 Plackett-Burman (PB)试验设计与分析
根据上述单因素试验结果设计了PB试验,以A-浓度(mg/L)、B-接种量(%)、C-盐度(‰)、D-pH、E-温度(℃)为影响因素,通过Design-Expert 13生成了具包含12条试验设计的正交矩阵,见
Test number | A-concentration (mg/L) | B | C- salinity (‰) | D- pH | E- T/℃ | 48 h removal efficiency (%) |
---|---|---|---|---|---|---|
1 | 50 | 7 | 20 | 9.0 | 35 | 54.48 |
2 | 10 | 7 | 35 | 5.0 | 35 | 31.56 |
3 | 50 | 3 | 35 | 9.0 | 25 | 43.31 |
4 | 10 | 7 | 20 | 9.0 | 35 | 41.31 |
5 | 10 | 3 | 35 | 5.0 | 35 | 30.93 |
6 | 10 | 3 | 20 | 9.0 | 25 | 35.11 |
7 | 50 | 3 | 20 | 5.0 | 35 | 39.05 |
8 | 50 | 7 | 20 | 5.0 | 25 | 44.09 |
9 | 50 | 7 | 35 | 5.0 | 25 | 37.32 |
10 | 10 | 7 | 35 | 9.0 | 25 | 32.38 |
11 | 50 | 3 | 35 | 9.0 | 35 | 48.87 |
12 | 10 | 3 | 20 | 5.0 | 25 | 26.85 |
通过回归分析得到多元一次回归方程(以编码值表示),如
Y48 h降解率=38.77+5.75A+1.42B–1.38C+
3.81D+2.26E | (2) |
回归方程拟合分析结果表明,Fisher检验(F值)为21.65,对应的概率检验(P值)为0.000 9,远小于系统所给的参考值0.001,表明模型整体在统计上极显著。通过
2.4.2 响应面优化试验
筛选出A-浓度、D-pH、E-温度作为48 h降解率条件中的关键影响因素。分别取3个因素的高(+1)、中(0)、低(‒1) 3个水平,设计Box-Behnken响应面试验,见
Test number | A- concentration (mg/L) | B- pH | C- T/℃ | 48 h removal efficiency (%) |
---|---|---|---|---|
1 | 10 | 5.0 | 30 | 34.68 |
2 | 50 | 5.0 | 30 | 32.82 |
3 | 10 | 9.0 | 30 | 30.19 |
4 | 50 | 9.0 | 30 | 50.59 |
5 | 10 | 7.0 | 25 | 28.36 |
6 | 50 | 7.0 | 25 | 43.27 |
7 | 10 | 7.0 | 35 | 38.15 |
8 | 50 | 7.0 | 35 | 40.12 |
9 | 30 | 5.0 | 25 | 41.47 |
10 | 30 | 9.0 | 25 | 42.41 |
11 | 30 | 5.0 | 35 | 43.74 |
12 | 30 | 9.0 | 35 | 46.73 |
13 | 30 | 7.0 | 30 | 60.72 |
14 | 30 | 7.0 | 30 | 58.92 |
15 | 30 | 7.0 | 30 | 60.78 |
16 | 30 | 7.0 | 30 | 57.05 |
17 | 30 | 7.0 | 30 | 59.98 |
48 h降解率模型建立与分析得到二次多项回归模型,如
Y48 h降解率=59.49+4.43A+2.15B+1.65C+5.56AB–
3.24AC+0.51BC–14.27 | (3) |
方程中的A、B和C分别代表浓度(mg/L)、pH和温度(℃)的编码值。回归方程中系数的正负符号表示变量的协同效应和拮抗效应。由
经过实验数据与二次多项式回归模型的统计检验可知,响应值F值为71.10,概率值(P值)<0.000 1,这表明由于噪声引起模型误差的可能性仅为0.01
如

图5 影响条件对48 h降解率的响应曲面图和等高线图
Figure 5 The response surface and contour plots of the 48 hour degradation efficiency under different influencing conditions. A: pH and concentration; B: Temperature and concentration; C: Temperature and pH.
2.4.3 优化结果验证
为了验证拟合模型对SMX降解率最大值的可信度,按照预测的最大降解率条件(浓度34 mg/L、pH 7.4、温度30 ℃),进行了3个平行实验。3个重复实验的最大降解率为(59.26±2.00)%。预测结果与实验结果之间具有良好的一致性。
2.5 降解产物及降解途径分析
SMX的分子结构中,五元环与磺胺基团相连,较小的角张力赋予其相对稳定的分子构

图6 SMX降解产物及降解路径推断
Figure 6 SMX degradation products and proposed degradation pathway.
2.6 急性毒性试验
采用发光细菌法对不同浓度的样品进行水质急性毒性检测,评估样品对发光细菌的毒性效应。设置了5、10、20、50、100 mg/L 5个SMX浓度梯度,并在0、24、48 h 3个关键时间点进行取样检测。结果如

图7 水质急性毒性实验
Figure 7 Water quality acute toxicity test. *: P<0.05 vs. 0 h untreated group. +: P<0.05 vs. 24 h treatment group.
3 结论
从海水养殖池底泥中筛选出了一株具有耐盐性能的SMX降解菌LS-1。通过16S rRNA基因测序和系统发育树分析,鉴定该菌株为产碱杆菌(Alcaligenes sp.)。菌株的对数生长期在28 h左右,在48 h内对SMX的降解率达到52.13%。单因素试验表明,菌株在不同盐度(15‰-35‰)、污染物浓度(10-100 mg/L)、温度(20-35 ℃)和初始pH (5.0-9.0)条件下表现出良好的适应性。对SMX降解率影响显著的因素从强到弱排序为:SMX浓度、pH和温度。响应面分析显示,SMX浓度与pH、浓度和温度之间的交互作用显著。SMX降解的最优条件为:SMX浓度 34 mg/L、初始pH 7.4、温度30 ℃,在此条件下,降解率可达59.26%,经验证实际值与预测值具有良好的一致性。经质谱分析,LS-1菌对SMX的降解产物为N-乙酰基-SMX (m/z=295)、3A5MI和磺胺甲恶唑羟胺(m/z=269)等,主要降解途径为氨基乙酰化,羟基化和五元环N-O键断裂。在SMX低浓度时,经过LS-1菌的处理,水质毒性迅速减弱。该菌株能够有效控制海水养殖环境中的SMX,对抗生素污染的防治具有很好的应用前景。
作者贡献声明
厉怡君:实验设计、操作及论文撰写;赵阳国:实验指导、论文修改和润色;刘磊:协助实验操作、论文讨论;岳梦晨:协助实验操作、论文讨论;张彦超:协助实验操作及指导,李欢欢:协助实验操作。
利益冲突
作者声明不存在任何可能会影响本文所报告工作的已知经济利益或个人关系。
参考文献
CHEN GL, LI YZ, WANG J. Occurrence and ecological impact of microplastics in aquaculture ecosystems[J]. Chemosphere, 2021, 274: 129989. [百度学术]
NAYLOR RL, HARDY RW, BUSCHMANN AH, BUSH SR, CAO L, KLINGER DH, LITTLE DC, LUBCHENCO J, SHUMWAY SE, TROELL M. A 20-year retrospective review of global aquaculture[J]. Nature, 2021, 591(7851): 551-563. [百度学术]
HE LX, HE LY, GAO FZ, ZHANG M, CHEN J, JIA WL, YE P, JIA YW, HONG B, LIU SS, LIU YS, ZHAO JL, YING GG. Mariculture affects antibiotic resistome and microbiome in the coastal environment[J]. Journal of Hazardous Materials, 2023, 452: 131208. [百度学术]
ZHANG SQ, ZHAO X, FENG KS, HU YC, TILLOTSON MR, YANG L. Do mariculture products offer better environment and nutritional choices compared to land-based protein products in China?[J]. Journal of Cleaner Production, 2022, 372: 133697. [百度学术]
GONZÁLEZ-GAYA B, GARCÍA-BUENO N, BUELOW E, MARIN A, RICO A. Effects of aquaculture waste feeds and antibiotics on marine benthic ecosystems in the Mediterranean Sea[J]. Science of the Total Environment, 2022, 806: 151190. [百度学术]
彭霈. 我国海水养殖污染问题与其治理研究[D]. 青岛: 中国海洋大学硕士学位论文, 2014. [百度学术]
PENG P. The Study of pollution and control in saltwater aquaculture in China[D]. Qingdao: Master’s Thesis of Ocean University of China, 2014 (in Chinese). [百度学术]
于世雄. 基于生命周期的海水养殖环境影响评估: 以山东省为例[D]. 济南: 山东大学硕士学位论文, 2023. [百度学术]
YU SX. Assessment of environmental effect of seawater aquaculture based on LCA: taking Shandong Province as an example[D].Jinan: Master’s Thesis of Shandong University, 2023 (in Chinese). [百度学术]
CHEN JJ, GAO MC, ZHAO YG, GUO L, JIN CJ, JI JY, SHE ZL. Nitrogen and sulfamethoxazole removal in a partially saturated vertical flow constructed wetland treating synthetic mariculture wastewater[J]. Bioresource Technology, 2022, 358: 127401. [百度学术]
CHEN JM, SUN RX, PAN CG, SUN Y, MAI BX, LI QX. Antibiotics and food safety in aquaculture[J]. Journal of Agricultural and Food Chemistry, 2020, 68(43): 11908-11919. [百度学术]
LIU XW, LV K, DENG CX, YU ZM, SHI JH, JOHNSON AC. Persistence and migration of tetracycline, sulfonamide, fluoroquinolone, and macrolide antibiotics in streams using a simulated hydrodynamic system[J]. Environmental Pollution, 2019, 252: 1532-1538. [百度学术]
ZHANG B, HE YK, SHI WX, LIU LJ, LI L, LIU C, LENS PNL. Biotransformation of sulfamethoxazole (SMX) by aerobic granular sludge: Removal performance, degradation mechanism and microbial response[J]. Science of the Total Environment, 2023, 858: 159771. [百度学术]
DENG YY, ZOU MY, LIU W, LIAN YL, GUO QM, ZHANG XM, DAN A. Antibiotic removal and microbial response mechanism in constructed wetlands treating aquaculture wastewater containing veterinary drugs[J]. Journal of Cleaner Production, 2023, 394: 136271. [百度学术]
LIANG DH, HU YY, LIANG DM, CHENGA JH, CHENA YC. Bioaugmentation of moving bed biofilm reactor (MBBR) with Achromobacter JL9 for enhanced sulfamethoxazole (SMX) degradation in aquaculture wastewater[J]. Ecotoxicology and Environmental Safety, 2021, 207: 111258. [百度学术]
BORSETTO C, RAGUIDEAU S, TRAVIS E, KIM DW, LEE DH, BOTTRILL A, STARK R, SONG LJ, CHA CJ, PEARSON J, QUINCE C, SINGER AC, WELLINGTON EMH. Impact of sulfamethoxazole on a riverine microbiome[J]. Water Research, 2021, 201: 117382. [百度学术]
PRASANNAMEDHA G, KUMAR PS. A review on contamination and removal of sulfamethoxazole from aqueous solution using cleaner techniques: present and future perspective[J]. Journal of Cleaner Production, 2020, 250: 119553. [百度学术]
WANG QN, WANG HD, LV M, WANG XY, CHEN LX. Sulfamethoxazole degradation by Aeromonas caviae and co-metabolism by the mixed bacteria[J]. Chemosphere, 2023, 317: 137882. [百度学术]
KONG SJ, ZHAO YG, GUO L, GAO MC, JIN CJ, SHE ZL. Transcriptomics of Planococcus kocurii O516 reveals the degrading metabolism of sulfamethoxazole in marine aquaculture wastewater[J]. Environmental Pollution, 2020, 265: 114939. [百度学术]
LIU XH, CHEN J, LIU Y, WAN ZF, GUO XC, LU SY, QIU DR. Sulfamethoxazole degradation by Pseudomonas silesiensis F6a isolated from bioelectrochemical technology-integrated constructed wetlands[J]. Ecotoxicology and Environmental Safety, 2022, 240: 113698. [百度学术]
LI YX, YANG T, LIN XJ, HUANG JF, ZENG JW, CAI QY, ZHANG YL, RONG JN, YU WD, QIU JR, PANG YW, ZHOU JL. Isolation, identification, and optimization of conditions for the degradation of four sulfonamide antibiotics and their metabolic pathways in Pseudomonas stutzeri strain DLY-21[J]. Heliyon, 2024, 10(7): e29123. [百度学术]
QI MY, MA XD, LIANG B, ZHANG LY, KONG DY, LI ZL, WANG AJ. Complete genome sequences of the antibiotic sulfamethoxazole-mineralizing bacteria Paenarthrobacter sp. P27 and Norcardiodes sp. N27[J]. Environmental Research, 2022, 204: 112013. [百度学术]
NODEHI RN, SHEIKHI S. Nanomaterial-based AOPs for the removal of organic pollutants in aqueous matrices: a systematic review of response surface methodology (RSM) models[J]. Environmental Technology & Innovation, 2024, 35: 103718. [百度学术]
SHAMSKILANI M, NIAVOL KP, NABAVI E, MEHRNIA MR, SHARAFI AH. Removal of emerging contaminants in a membrane bioreactor coupled with ozonation: optimization by response surface methodology (RSM)[J]. Water, Air, & Soil Pollution, 2023, 234(5): 304. [百度学术]
CHEN JL, XU J, ZHANG SN, LIU F, PENG JW, PENG YX, WU JS. Nitrogen removal characteristics of a novel heterotrophic nitrification and aerobic denitrification bacteria, Alcaligenes faecalis strain WT14[J]. Journal of Environmental Management, 2021, 282: 111961. [百度学术]
CAO XH, ZHAO BH, WU YM, HUANG J, WANG HZ, SUN XY, LI SJ. Characterization of Alcaligenes aquatilis as a novel member of heterotrophic nitrifier-aerobic denitrifier and its performance in treating piggery wastewater[J]. Bioresource Technology, 2022, 354: 127176. [百度学术]
AO XW, LIU WJ, SUN WJ, YANG C, LU ZD, LI C. Mechanisms and toxicity evaluation of the degradation of sulfamethoxazole by MPUV/PMS process[J]. Chemosphere, 2018, 212: 365-375. [百度学术]
DŁUGOSZ M, PAWEŁ Ż, KWIECIEŃ A, SZCZUBIAŁKA K, KRZEK J, NOWAKOWSKA M. Photocatalytic degradation of sulfamethoxazole in aqueous solution using a floating TiO2-expanded perlite photocatalyst[J]. Journal of Hazardous Materials, 2015, 298: 146-153. [百度学术]
URBANO M. Factors controlling Macondo oil biodegraqdation on a rapidly eroding coastal headlands beach[D]. Louisiana State University Libraries, Master of Science in Civil Engineering (MSCE), 2012. DOI:10.31390/gradschool_theses.3748. [百度学术]
XIONG Q, LIU YS, HU LX, SHI ZQ, CAI WW, HE LY, YING GG. Co-metabolism of sulfamethoxazole by a freshwater microalga Chlorella pyrenoidosa[J]. Water Research, 2020, 175: 115656. [百度学术]
SHI JX, XU CY, HAN YX, HAN HJ. Enhanced anaerobic degradation of nitrogen heterocyclic compounds with methanol, sodium citrate, Chlorella, Spirulina, and carboxymethylcellulose as co-metabolic substances[J]. Journal of Hazardous Materials, 2020, 384: 121496. [百度学术]
VO HNP, NGO HH, GUO WS, LIU YW, CHANG SW, NGUYEN DD, ZHANG XB, LIANG H, XUE S. Selective carbon sources and salinities enhance enzymes and extracellular polymeric substances extrusion of Chlorella sp. for potential co-metabolism[J]. Bioresource Technology, 2020, 303: 122877. [百度学术]
MISHRA A, MANDOLI A, JHA B. Physiological characterization and stress-induced metabolic responses of Dunaliella salina isolated from salt pan[J]. Journal of Industrial Microbiology & Biotechnology, 2008, 35(10): 1093-1101. [百度学术]
XIONG JQ, KURADE MB, PATIL DV, JANG M, PAENG KJ, JEON BH. Biodegradation and metabolic fate of levofloxacin via a freshwater green Alga, Scenedesmus obliquus in synthetic saline wastewater[J]. Algal Research, 2017, 25: 54-61. [百度学术]
REIS PJM, REIS AC, RICKEN B, KOLVENBACH BA, MANAIA CM, CORVINI PFX, NUNES OC. Biodegradation of sulfamethoxazole and other sulfonamides by Achromobacter denitrificans PR1[J]. Journal of Hazardous Materials, 2014, 280: 741-749. [百度学术]
YAN RF, WANG YB, LI JH, WANG XH, WANG YK. Determination of the lower limits of antibiotic biodegradation and the fate of antibiotic resistant genes in activated sludge: both nitrifying bacteria and heterotrophic bacteria matter[J]. Journal of Hazardous Materials, 2022, 425: 127764. [百度学术]
MOHAPATRA B, PHALE PS. Microbial degradation of naphthalene and substituted naphthalenes: metabolic diversity and genomic insight for bioremediation[J]. Frontiers in Bioengineering and Biotechnology, 2021, 9: 602445. [百度学术]
GOERS L, FREEMONT P, POLIZZI KM. Co-culture systems and technologies: taking synthetic biology to the next level[J]. Journal of the Royal Society, Interface, 2014, 11(96): 20140065. [百度学术]
赵栩宁, 马冬雪, 赵阳国. 海水养殖生境中硫酸盐还原菌活性的抑制机制[J]. 微生物学报, 2022, 62(8): 3048-3061. [百度学术]
ZHAO XN, MA DX, ZHAO YG. Sulfide-producing activity inhibition of sulfate-reducing bacteria in mariculture[J]. Acta Microbiologica Sinica, 2022, 62(8): 3048-3061 (in Chinese). [百度学术]
ISLAM MS. Nitrogen and phosphorus budget in coastal and marine cage aquaculture and impacts of effluent loading on ecosystem: review and analysis towards model development[J]. Marine Pollution Bulletin, 2005, 50(1): 48-61. [百度学术]
NAM SN, CHO H, HAN J, HER N, YOON J. Photocatalytic degradation of acesulfame K: optimization using the Box-Behnken design (BBD)[J]. Process Safety and Environmental Protection, 2018, 113: 10-21. [百度学术]
张晓菲, 倪晓菁, 张子仪, 等. 响应面法优化黑曲霉固定化条件及其对土壤中溴氰菊酯的降解特性研究[J]. 微生物学报, 2023, 63(12): 4574-4593. [百度学术]
ZHANG XF. NI XQ. ZHANG ZY. PRNG FQ. ZHOU SY. RAN XY. FANG YW. LIANG XY. ZHOU TY. WANG Q. LIU P. Optimization of immobilization conditions of Aspergillus niger for degrading deltamethrin in soil[J]. Acta Microbiologica Sinica, 2023, 63(12): 4574-4593 (in Chinese). [百度学术]
FAVIER L, ANDREI-IONUȚ SIMION, HLIHOR RM, FEKETE-KERTÉSZ I, MOLNÁR M, HARJA M, VIAL C. Intensification of the photodegradation efficiency of an emergent water pollutant through process conditions optimization by means of response surface methodology[J]. Journal of Environmental Management, 2023, 328: 116928. [百度学术]
WANG ZY, SONG W, LI J, ZHANG XL, WANG HJ. Optimization and mechanism of Tetrabromobisphenol A removal by dithionite under anaerobic conditions: Response surface methodology and degradation pathway[J]. Journal of Environmental Management, 2022, 321: 116034. [百度学术]
CHEN CY, LUO ZF, TU HX, LIN XJ, PANG YW, HUANG JF, ZHANG J, WANG XJ, CAI QY, WEI ZB, ZENG JW, QIU JR. Response surface methodology and Box-Behnken design optimization of Sulfaquinoxaline removal efficiency and degradation mechanisms by Bacillus sp. strain DLY-11[J]. Journal of Hazardous Materials, 2025, 486: 136986. [百度学术]
TIAN SH, YOU LL, HUANG X, LIU CX, SU JQ. Efficient sulfamethoxazole biotransformation and detoxification by newly isolated strain Hydrogenophaga sp. SNF1 via a ring ortho-hydroxylation pathway[J]. Journal of Hazardous Materials, 2024, 480: 136113. [百度学术]
ZHANG MY, FAN DP, SU C, PAN LQ, HE QL, LI ZL, LIU C. Biotransformation of sulfamethoxazole by a novel strain, Nitratireductor sp. GZWM139: characterized performance, metabolic mechanism and application potential[J]. Journal of Hazardous Materials, 2023, 441: 129861. [百度学术]
DONG ZK, YAN XJ, WANG JH, ZHU LS, WANG J, LI CY, ZHANG WJ, WEN SF, KIM YM. Mechanism for biodegradation of sulfamethazine by Bacillus cereus H38[J]. Science of the Total Environment, 2022, 809: 152237. [百度学术]
HARNISCH F, GIMKIEWICZ C, BOGUNOVIC B, KREUZIG R, SCHRÖDER U. On the removal of sulfonamides using microbial bioelectrochemical systems[J]. Electrochemistry Communications, 2013, 26: 77-80. [百度学术]
RADKE M, LAUWIGI C, HEINKELE G, MÜRDTER TE, LETZEL M. Fate of the antibiotic sulfamethoxazole and its two major human metabolites in a water sediment test[J]. Environmental Science & Technology, 2009, 43(9): 3135-3141. [百度学术]
CRIBB AE, NAKAMURA H, GRANT DM, MILLER MA, SPIELBERG SP. Role of polymorphic and monomorphic human arylamine N-acetyltransferases in determining sulfamethoxazole metabolism[J]. Biochemical Pharmacology, 1993, 45(6): 1277-1282. [百度学术]
RICKEN B, KOLVENBACH BA, BERGESCH C, BENNDORF D, KROLL K, STRNAD H, VLČEK Č, ADAIXO R, HAMMES F, SHAHGALDIAN P, SCHÄFFER A, KOHLER HE, CORVINI PF. FMNH2-dependent monooxygenases initiate catabolism of sulfonamides in Microbacterium sp. strain BR1 subsisting on sulfonamide antibiotics[J]. Scientific Reports, 2017, 7(1): 15783. [百度学术]
RICKEN B, CORVINI PFX, CICHOCKA D, PARISI M, LENZ M, WYSS D, MARTÍNEZ-LAVANCHY PM, MÜLLER JA, SHAHGALDIAN P, TULLI LG, KOHLER HE, KOLVENBACH BA. Ipso-hydroxylation and subsequent fragmentation: a novel microbial strategy to eliminate sulfonamide antibiotics[J]. Applied and Environmental Microbiology, 2013, 79(18): 5550-5558. [百度学术]
ZHAO X, PEI WN, QI YH, LI YB, KONG XQ. Enhanced aerobic granular sludge with micro-electric field for sulfamethoxazole degradation: efficiency, mechanism, and microbial community[J]. Chemosphere, 2024, 354: 141741. [百度学术]
QI MY, LIANG B, ZHANG L, MA XD, YAN L, DONG WC, KONG DY, ZHANG LY, ZHU HZ, GAO SH, JIANG JD, LIU SJ, CORVINI PF, WANG AJ. Microbial interactions drive the complete catabolism of the antibiotic sulfamethoxazole in activated sludge microbiomes[J]. Environmental Science & Technology, 2021, 55(5): 3270-3282. [百度学术]
REIS AC, KOLVENBACH BA, NUNES OC, CORVINI PFX. Biodegradation of antibiotics: The new resistance determinants–part I[J]. New Biotechnology, 2020, 54: 34-51. [百度学术]
MAJEWSKY M, WAGNER D, DELAY M, BRÄSE S, YARGEAU V, HORN H. Antibacterial activity of sulfamethoxazole transformation products (TPs): general relevance for sulfonamide TPs modified at the Para position[J]. Chemical Research in Toxicology, 2014, 27(10): 1821-1828. [百度学术]
WANG J, SHEN M, WANG HL, DU YS, ZHOU XQ, LIAO ZW, WANG HB, CHEN ZQ. Red mud modified sludge biochar for the activation of peroxymonosulfate: Singlet oxygen dominated mechanism and toxicity prediction[J]. Science of the Total Environment, 2020, 740: 140388. [百度学术]
WANG L, LIU YL, MA J, ZHAO F. Rapid degradation of sulphamethoxazole and the further transformation of 3-amino-5-methylisoxazole in a microbial fuel cell[J]. Water Research, 2016, 88: 322-328. [百度学术]