Diversity of antibiotic resistance genes and encoding ribosomal protection proteins gene in livestock waste polluted environment

The rapid development and increase of antibiotic resistance are global phenomena resulting from the extensive use of antibiotics in human clinics and animal feeding operations. Antibiotics can promote the occurrence of antibiotic resistance genes (ARGs), which can be transferred horizontally to humans and animals through water and the food chain. In this study, the presence and abundance of ARGs in livestock waste was monitored by quantitative PCR. A diverse set of bacteria and tetracycline resistance genes encoding ribosomal protection proteins (RPPs) from three livestock farms and a river were analyzed through denaturing gradient gel electrophoresis (DGGE). The abundance of sul(I) was 10³ to 10⁵ orders of magnitude higher than that of sul(II). Among 11 tet-ARGs, the most abundant was tet(O). The results regarding bacterial diversity indicated that the presence of antibiotics might have an evident impact on bacterial diversity at every site, particularly at the investigated swine producer. The effect of livestock waste on the bacterial diversity of soil was stronger than that of water. Furthermore, a sequencing analysis showed that tet(M) exhibited two genotypes, while the other RPPs-encoding genes exhibited at least three genotypes. This study showed that various ARGs and RPPs-encoding genes are particularly widespread among livestock.     

全氟辛烷磺酸(PFOS)对斑马鱼卵黄蛋白原mRNA水平的影响

为了研究环境低剂量全氟辛烷磺酸（perfluorooctane sulfonate,PFOS）对水生生物的内分泌干扰效应和可能的作用机制,测定了PFOS对斑马鱼（Brachydanio rerio）肝脏中卵黄蛋白原（vitellogenin,VTG）mRNA水平的影响.将斑马鱼暴露于4个PFOS的环境低剂量浓度组（0.1、1、10、100μg.L-1）中进行21d毒性试验,收集肝脏样品,提取RNA,采用荧光定量PCR（qRT-PCR）分别检测VTG1和VTG3的mRNA水平.结果表明：①PFOS暴露引起雄性斑马鱼肝脏VTG1和VTG3 mRNA水平升高,VTG1 mRNA水平升高与剂量呈正相关,在100μg.L-1暴露浓度处与对照组呈现显著性差异;VTG3的mRNA水平变化与剂量呈倒U型曲线,呈现典型的毒物刺激荷尔蒙效应,在10和100μg.L-1暴露浓度处与对照组呈现显著性差异;②PFOS暴露引起雌性斑马鱼肝脏中VTG1 mRNA水平升高,在10μg.L-1暴露浓度处与对照组呈现显著性差异,但在高浓度（10和100μg.L-1）处试验结果误差较大;VTG3 mRNA水平只在10μg.L-1暴露浓度处升高,但相比于对照组均没有显著性差异.试验结果表明PFOS暴露对斑马鱼的内分泌干扰作用明显,其毒性作用机制可能是类雌激素效应,而肝脏中VTG1和VTG3mRNA水平可能作为PFOS内分泌干扰效应评价的敏感生物标志物,但VTG1和VTG3 mRNA水平的响应曲线呈现基因亚型和性别差异.     

崇明东滩夏季沉积物厌氧氨氧化菌群落结构与空间分布特征

为探索长江口崇明东滩表层沉积物中是否存在厌氧氨氧化菌以及厌氧氨氧化菌的群落结构与空间分布特征,以崇明东滩高、中、低潮滩夏季表层沉积物中总DNA为模板,PCR扩增沉积物样品中厌氧氨氧化菌特异性16S rDNA片段,通过克隆、测序,构建相应基因文库,将文库中有效克隆序列分别提交到GenBank中进行相似序列搜索,并利用MEGA5绘制系统发育树.分析结果发现,克隆序列CM-L-7和CM-L-18与已发现厌氧氨氧化菌Candidatus"Scalindua sp."同源性达98%,CM-L-13与Candidatus"Scalindua wagneri"同源性达94%,CM-M-6与Candidatus"Kuenenia sp."同源性达94%,CM-M-22与Anaerobicammonium-oxidizing planctomycete JMK-1的同源性达95%,CM-H-15与Candidatus"Kuenenia stuttgartiensis"的同源性达94%.表明,崇明东滩高、中、低潮滩表层沉积物中均存在厌氧氨氧化菌,但所属菌属不同,低潮滩以Candidatus"Scalindua"为主,中、高潮滩以Candidatus"Kuenenia"为主.相比之下,中潮滩厌氧氨氧化菌群落结构较为复杂.部分克隆序列与已发现厌氧氨氧化菌存在较大进化距离,表明崇明东滩可能还存在其他具有潜在厌氧氨氧化功能的细菌.     

Real-time fluorescent quantitative immuno-PCR method for determination of fluoranthene in water samples with a molecular beacon

A reliable and sensitive competitive real-time fluorescent quantitative immuno-PCR (RTFQ-IPCR) assay using a molecular beacon was developed for the determination of trace fluoranthene (FL) in the environment. Under optimized assay conditions, FL can be determined in the concentration range from 1 fg/mL to 100 ng/mL, with y = 0.194x + 7.859, and a correlation coe cient of 0.967 was identified, with a detection limit of 0.6 fg/mL. Environmental water samples were successfully analyzed, recovery was between 90% and 116%, with intra-day relative standard deviation (RSD) of 6.7%–12.8% and inter-day RSD of 8.4%–15.2%. The results obtained from RTFQ-IPCR were confirmed by ELISA, showing good accuracy and suitability to analyze FL in field samples. As a highly sensitive method, the molecular beacon-based RTFQ-IPCR is acceptable and promising for providing reliable test results to make environmental decisions.     

Do land utilization patterns a ect methanotrophic communities in a Chinese upland red soil?

Soil samples were collected from three plots under di erent land utilization patterns including degradation, farming, and restoration. The abundances of methanotrophs were quantified using real-time polymerase chain reaction (PCR) based on the pmoA and 16S rRNA genes, and the community fingerprint was analyzed using denaturing gradient gel electrophoresis (DGGE) aiming at pmoA gene. Significantly lower 16S rRNA and pmoA genes copies were found in the degradation treatment than in farming and restoration. Higher abundances of Type I than those of Type II methanotrophs were detected in all treatments. The treatment of farming was clearly separated from degradation and restoration according to the DGGE profile by cluster analysis. The lowest diversity indices were observed in the F (farming plot), suggesting that the community structure was strongly a ected by farming activities. There were significantly positive correlations between the copy numbers of pmoA also Type II-related 16S rRNA genes and soil available K content. Strong negative and positive correlations were found between Type I and soil pH, and available P content, respectively. We concluded that the vegetation cover or not, soil characteristics including pH and nutrients of P and K as a result of anthropogenic disturbance may be key factors a ecting methanotrophic communities in upland soil.     

基于实时控制的智能控制在废水处理中的应用

针对废水处理系统具有时变性、非线性、复杂性和不确定性等特点,提出采用智能控制对其进行实时有效控制从而提高处理效率、降低能耗,对智能控制的主要分支模糊控制、神经网络控制和专家系统进行讨论,介绍总结了国内外专家学者应用智能控制技术实现水处理中实时控制的最新动态。结果表明,废水处理系统采用智能控制可以有效地实现实时控制,提高处理效率、降低能耗和成本。     

金鱼组织中壬基酚的含量检测新方法探讨

建立一种直接检测环境中痕量壬基酚的外切酶保护-荧光定量PCR方法。首先向含雌激素受体及相关蛋白的细胞溶质中加入壬基酚-丙酮溶液,使受体蛋白活化,从而在体外与含雌激素反应位点的双链结合DNA作用形成壬基酚-雌激素受体-DNA复合物。复合物用核酸外切酶和S1核酸酶消解,去除未受到蛋白质保护的结合DNA。将消解后产物作为模板,进行荧光定量PCR扩增反应,建立壬基酚浓度与标准DNA拷贝数的对数之间的线性关系为：y（壬基酚浓度的对数）=1.637x（标准DNA拷贝数的对数）-9.505。该法检出限为10^-8g/L,用该法对金鱼肝脏组织中壬基酚进行检测,添加回收率水平在98.5%112.2%,变异系数在4.3%以下。     

畜禽养殖中沙门氏菌污染的PCR快速检测

针对畜禽养殖是造成农村水环境生物性污染的主要原因,指出用常规生化检测技术耗时长、效率低,不能满足环境管理的要求：应用PCR技术检测沙门氏菌,特异性高、敏感性强、耗时短,与常规分离鉴定方法比较,完全符合实验证明,PCR技术可很好的应用于沙门氏菌污染的调查与监测。     

进水浓度或HRT变化时微好氧处理微生态机制研究

考察改变进水COD浓度（11 000、 15 000、 30 000 mg/Ｌ）或水力停留时间（HRT为42 h、 25 h）对模拟高浓度有机废水微好氧连续小试处理效果的影响,并采用荧光原位杂交-流式细胞术（FISH-FCM）、聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE) 等分子生物学技术及Biolog　FF微孔板法分析了不同处理阶段稳定期曝气柱污泥的微生态变化,以探讨进水COD浓度或HRT改变时微好氧处理效果变化的微生态机制.结果表明,在全部4个处理阶段,曝气柱污泥中酵母含量均保持在＞99.9％的水平.随着进水COD浓度的上升,污泥浓度由2.0 g/Ｌ上升到7.3 g/Ｌ,COD比去除速度由2.3 kg/（kg·d）下降到1.7 kg/（kg·d）,曝气柱污泥真菌群落结构多样性指数由2.05上升至2.19,代谢多样性指数由4.42上升至4.45;而随着HRT下降,污泥浓度从7.3 g/Ｌ下降至6.0 g/Ｌ,COD比去除速度从1.7 kg/（kg·d）上升至2.8 kg/（kg·d）,真菌群落结构多样性指数和代谢多样性指数则分别从2.19和4.45降至0.79和4.36.作为提高进水有机负荷的主要措施,提高进水COD浓度和缩短HRT对于废水处理效果和曝气柱微生态存在截然相反的影响.