浮游动物DNA宏条形码标志基因比较研究

DNA宏条形码技术作为一种新型生物监测方法,在未来生态环境监测中有巨大的应用潜力。目前,浮游动物DNA宏条形码监测仍在发展阶段,需要首先对其(采样方法、引物选择和数据分析等)进行标准化和调整,然后才能用于常规流域生态监测。其中,如何选择合适的PCR扩增引物是DNA宏条形码生物监测标准化的关键问题之一。本研究比较了COI、18SV9和16S通用引物在浮游动物DNA宏条形码监测中的差异,为初步建立规范化的浮游动物DNA宏条形码监测方法提供技术支撑。结果表明,16S引物对浮游动物具有更好的特异性,其产生的操作分类单元(operational taxonomic unit, OTU)有88.1%属于浮游动物。虽然18SV9引物具有更高的物种覆盖度,不仅能扩增出浮游动物,还能扩增出大量藻类和真菌,但其物种识别敏感性较差,不适合浮游动物物种水平多样性监测。COI引物的浮游动物物种特异性、物种覆盖度和物种识别敏感性都适中,检出的浮游动物物种数量高于18SV9引物和16S引物,更加适合浮游动物DNA宏条形码监测。     

Biofiltration and disinfection codetermine the bacterial antibiotic resistome in drinking water: A review and meta-analysis

• Published data was used to analyze the fate of ARGs in water treatment. • Biomass removal leads to the reduction in absolute abundance of ARGs. • Mechanism that filter biofilm maintain ARB/ARGs was summarized. • Potential BAR risks caused by biofiltration and chlorination were proposed. The bacterial antibiotic resistome (BAR) is one of the most serious contemporary medical challenges. The BAR problem in drinking water is receiving growing attention. In this study, we focused on the distribution, changes, and health risks of the BAR throughout the drinking water treatment system. We extracted the antibiotic resistance gene (ARG) data from recent publications and analyzed ARG profiles based on diversity, absolute abundance, and relative abundance. The absolute abundance of ARG was found to decrease with water treatment processes and was positively correlated with the abundance of 16S rRNA (r² = 0.963, p<0.001), indicating that the reduction of ARG concentration was accompanied by decreasing biomass. Among treatment processes, biofiltration and chlorination were discovered to play important roles in shaping the bacterial antibiotic resistome. Chlorination exhibited positive effects in controlling the diversity of ARG, while biofiltration, especially granular activated carbon filtration, increased the diversity of ARG. Both biofiltration and chlorination altered the structure of the resistome by affecting relative ARG abundance. In addition, we analyzed the mechanism behind the impact of biofiltration and chlorination on the bacterial antibiotic resistome. By intercepting influent ARG-carrying bacteria, biofilters can enrich various ARGs and maintain ARGs in biofilm. Chlorination further selects bacteria co-resistant to chlorine and antibiotics. Finally, we proposed the BAR health risks caused by biofiltration and chlorination in water treatment. To reduce potential BAR risk in drinking water, membrane filtration technology and water boiling are recommended at the point of use.     

Validation of Bacteroidales-based microbial source tracking markers for pig fecal pollution and their application in two rivers of North China

• Pig feces is the predominant excrement produced by animal husbandry in China. • The PF, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan MST assays showed better performance. • The pig-specific MST assays can contribute to managing the pig fecal pollution. In China, pig feces is the predominant source of excrement produced by animal husbandry. Improper use or direct discharge of pig feces can result in contamination of natural water systems. Microbial source tracking (MST) technology can identify the sources of fecal pollution in environmental water, and contribute to the management of pig fecal pollution by local environmental protection agencies. However, the accuracy of such assays can be context-dependent, and they have not been comprehensively evaluated under Chinese conditions. We aimed to compare the performance of five previously reported pig-specific MST assays (PF, Pig-Bac1^SYBR, Pig-Bac2^SYBR, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan, which are based on Bacteroidales 16S rRNA gene markers) and apply them in two rivers of North China. We collected a total of 173 fecal samples from pigs, cows, goats, chickens, humans, and horses across China. The PF assay optimized in this study showed outstanding qualitative performance and achieved 100% specificity and sensitivity. However, the two SYBR green qPCR assays (Pig-Bac1^SYBR and Pig-Bac2^SYBR) cross-reacted with most non-pig fecal samples. In contrast, both the Pig-1-Bac^TaqMan and Pig-2-Bac^TaqMan assays gave 100% specificity and sensitivity. Of these, the Pig-2-Bac^TaqMan assay showed higher reproducibility. Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand, Japan, and America. Using the PF and Pig-2-Bac^TaqMan assays, a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.     

Bioanalytical and instrumental analysis of thyroid hormone disrupting compounds in water sources along the Yangtze River

Thyroid hormone (TH) agonist and antagonist activities of water sources along the Yangtze River in China were surveyed by a green monkey kidney fibroblast (CV-1) cell-based TH reporter gene assay. Instrumental analysis was conducted to identify the responsible thyroid-active compounds. Instrumentally derived l-3,5,3′-triiodothyronine (T₃) equivalents (T₃-EQs) and thyroid receptor (TR) antagonist activity equivalents referring to dibutyl phthalate (DBP-EQs) were calculated from the concentrations of individual congeners. The reporter gene assay demonstrated that three out of eleven water sources contained TR agonist activity equivalents (TR-EQs), ranging from 286 to 293 ng T₃/L. Anti-thyroid hormone activities were found in all water sources with the TR antagonist activity equivalents referring to DBP (Ant-TR-EQs), ranging from 51.5 to 555.3 μg/L. Comparisons of the equivalents from instrumental and biological assays suggested that high concentrations of DBP and di-2-ethylhexyl phthalate (DEHP) were responsible for the observed TR antagonist activities at some locations along the Yangtze River.     

Use of reporter-gene based bacteria to quantify phenanthrene biodegradation and toxicity in soil

A phenanthrene-degrading bacterium, Sphingomonas paucimobilis EPA505 was used to construct two fluorescence-based reporter strains. Strain D harboring gfp gene was constructed to generate green fluorescence when the strain started to biodegrade phenanthrene. Strain S possessing gef gene was designed to die once phenanthrene biodegradation was initiated and thus to lose green fluorescence when visualized by a live/dead cell staining. Confocal laser scanning microscopic observation followed by image analysis demonstrates that the fluorescence intensity generated by strain D increased and the intensity by strain S decreased linearly at the phenanthrene concentration of up to 200 mg/L. Such quantitative increase and decrease of fluorescence intensity in strain D (i.e., from 1 to 11.90 ± 0.72) and strain S (from 1 to 0.40 ± 0.07) were also evident in the presence of Ottawa sand spiked with the phenanthrene up to 1000 mg/kg. The potential use of the reporter strains in quantitatively determining biodegradable or toxic phenanthrene was discussed.     

企业生产安全事故的“基因”分析

思索生产安全事故频发共性表象"违法、违章"背后的原因,提出"生产安全事故基因"概念。分析得出"去小概率性"、"社会责任缺失"和"非货币化产出分析能力短缺"3种生产安全事故基因,前两种基因是不可去基因,后一种是可去基因,三者会在不同的条件下以显性或隐性的形式呈现,并作用于生产过程;提出企业组织安全进化的常规对策——完善市场经济环境,创新对策——教育要为"安全"生产服务,强调学历教育系统在创新对策中的重要地位和作用。通过"基因"分析及其对策,为安全生产培养安全管理人员和相关人才,推动安全生产,实现经济与社会和谐发展。     

Bioreduction of hexavalent chromium by Exiguobacterium indicum strain MW1 isolated from marine water of Paradip Port,Odisha, India

Hexavalent chromium-tolerant (1500?mg/L) bacterium MW1 was isolated from harbour water of Paradip Port and evaluated for Cr(VI) reduction potential. The isolate was identified as Exiguobacterium indicum by biochemical and 16S rRNA gene sequence methods. Salt tolerance of the bacterium was evaluated in a wide range of NaCl concentrations (0.5–13%, w/v). The Cr(VI) reduction of the strain was evaluated and optimised with varied Cr(VI) concentrations (100–1000?mg/L), pH (5.0–9.0), temperature (30–40°C) and shaking velocity (100–150?rpm) in two different minimal media (M9 and Acetate). Under optimised conditions, after 192?h of incubation nearly 92%, 50% and 46% reduction in the M9 minimal medium and 91%, 47% and 40% reduction in the acetate minimal medium were observed for 100, 500 and 1000?mg/L of Cr(VI), respectively. The exponential rate equation for Cr(VI) reduction yielded higher rate constant value, that is, 1.27?×?10^?2?h^?1 (M9) and 1.17?×?10^?2?h^?1 (Acetate) in case of 100?mg/L and became lower for 500 and 1000?mg/L Cr(VI) concentrations. Further, the association of bacterial cells with reduced product was ascertained by Fourier transform infrared spectrometer, UV–Vis–DRS and field-emission scanning electron microscope–energy-dispersive X-ray analyses. The above study suggests that the higher reducing ability of the marine bacterium E. indicum MW1 will be suitable for Cr(VI) reduction from saline effluents.     

Telomere length in workers was effected by omethoate exposure and interaction between smoking and p21 polymorphisms

Abstract

Omethoate is an organophosphorus pesticide that poses a major health hazard, especially DNA damage. The purpose of this study was to investigate the factors affecting telomere length in workers exposed to omethoate by analyzing the interaction between cell cycle gene polymorphism and environmental factors. The exposure group consisted of 118 workers exposed to omethoate for 8–10?years, the control group comprised 115 healthy people without occupational toxicant exposure history. The telomere length of genomic DNA from peripheral blood leucocyte was determined with real-time PCR. Polymerase chain reaction and restriction fragment length polymorphism was used to detect the polymorphisms in p53, p21 and MDM2 gene. The telomere length in the (CA?+?AA) genotypes for p21 rs1801270 polymorphism was longer than that in the CC genotype in control group (P?=?0.015). The generalized linear model analysis indicated the interaction of the p21 rs1801270 polymorphic (CA?+?AA) genotypes and smoking has a significant effect on telomere length (β = ?0.258, P?=?0.085). The prolongation of telomere length in omethoate-exposed workers was associated with genotypes (CA?+?AA) of p21 rs1801270, and interactions of (CA?+?AA) genotypes and smoking factor.     

超强静磁场对染料脱色菌Escheriehia coli的acpD基因的影响

将大肠杆菌E.coli ATCC8739置于12.0 T超强静磁场(ultra-strong static magnetic field,SMF)中进行处理,获得了磁场处理0.5、1、2、4和8 h的菌株E.coli-SMFn(n=0.5、1、2、4、8)。在37℃、pH 7、静置的条件下,菌株对偶氮染料AR14(I.C.Acid Red 14)的脱色结果指出,当反应进行到4、6和8 h时,E.coli-SMF8的脱色效率分别为18%、55%和96%,远高于E.coli ATCC8739的3%、19%和50%,表明SMF作用显著地增强了菌株的脱色效率。基因组DNA提取、PCR扩增、分子克隆以及基因测序的实验结果进一步表明,全部6例E.coli ATCC8739菌株的偶氮还原酶基因(acpD)序列均与GenBank中报道的完全一致;而E.coli-SMF8菌株的acpD-SMF8核酸序列中缺失了第99位的胞嘧啶。该缺失突变不仅使acpD-SMF8的核酸序列与acpD的存在显著不同,同时得到了具新活性中心的偶氮还原酶AzoR-SMF8。这个新的活性中心具有更强的黄素(FMN)结合能力,因此使该酶与偶氮染料的亲和力大大增加,促进了脱色效率的提高。