磺胺类化合物对大肠杆菌突变QSAR预测模型初探

抗生素被广泛应用于医疗、农业和畜牧业等领域,但是长期滥用会促进细菌的突变作用。本文以大肠杆菌(E.coli)为受试生物,测定了10种磺胺类抗生素(SAs)单一暴露时对E.coli的突变效应,采用物化参数Ebinding(抗生素与其靶蛋白的相互作用能力)与突变效应参数lgRC0-2(最高可观测突变促进效应浓度)或lgRCmax(突变促进率最大值对应的浓度),建立了突变效应的QSAR模型,并采用雷达图进行验证。结果表明,lgRC0-2与Ebinding模型和lgRCmax与Ebinding、DMG(偶极矩)模型的拟合系数R2分别是0.888、0.873,即lgRC0-2或lgRCmax与Ebinding相关性均较好,可能由于磺胺类化合物(SAs)会作用于叶酸合成通路,影响嘌呤、嘧啶碱基的合成,从而对E.coli的突变具有促进效应,且雷达图验证表明,上述2个模型均具有良好的内部预测能力。本研究有望为抗生素使用带来的生态风险评价以及药物设计提供相关指导。     

Release,Dispersion, and Resuspension of Escherichia coli From Direct Fecal Deposits Under Controlled Flows1

Abstract: Water‐quality standards have been placed on fecal indicator organisms such as Escherichia coli in an attempt to limit the concentrations in water bodies. Cattle can be a significant source of bacteria to water systems, particularly when they are allowed direct access to streams. A flume study was conducted to quantify the effect and understand the transport of E. coli from directly deposited cattle manure. Five steady‐state flows, ranging from 0.00683 to 0.0176 m³/s, were studied and loads from a single cowpie exceeded the U.S. Environmental Protection Agency’s recommended water‐quality standards (235 CFU/100 ml) at each flow over the hour study period. Average E. coli concentrations ranged from 10² to 10⁵ CFU/100 ml over the hour sampling period for all flows. High spatial variations in E. coli concentrations were often seen at each sampling time, with higher concentrations typically at the bottom of the flume. E. coli resuspension was initially greater at 0.5 min after deposition, for the lowest flow (10⁵ CFU/m²/s); however, resuspension rates became similar over time, on the order of 10³ CFU/m²/s. This study demonstrates that the concentrations of E. coli can vary over the water column, and therefore grab samples may inaccurately measure bacteria concentrations and loads in streams. In addition, resuspension rates were often high, so the incorporation of this process into water‐quality models is important for bacteria prediction.     

远程氧等离子体灭活大肠杆菌的研究

研究放电等离子体灭活大肠杆菌的效果。采用远程氧等离子体反应器,分别考察探究放电功率、处理时间、氧气流量、远程距离及有机污染物对灭菌效果的影响,并观察灭菌前后大肠杆菌的显微结构。结果表明在100W,60s,40cm3/min条件下,远程氧等离子体的杀菌效果值在距放电区0、20、40cm处分别为3.42、3.38和3.32。有机物浓度升高,灭菌效果急剧下降。结论为氧等离子体在距放电区040cm内均能有效灭活大肠杆菌,但有机污染物影响较大。氧等离子体场中高能粒子的刻蚀作用是导致细菌死亡的主要原因。     

Biodegradation of azo dyes by genetically engineered azoreductase

A azoreductase gene with 537 bp was obtained by PCR amplification from Rhodobacter sphaeroides ASI. 1737. The enzyme,with a molecular weight of 18.7 kD, was efficiently expressed in Escherichia coli and its biodegradation characteristics for azo dyes were investigated. Furthermore, the reaction kinetics and mechanism of azo dyes catalyzed by the genetically engineered azoreductase were studied in detail. The presence of a hydrazo-intermediate was identified, which provided a convincing evidence for the assumption that azo dyes were degraded via an incomplete reduction stage.     

大肠杆菌总DNA快速提取方法的比较研究

从动物粪便中分离大肠埃希氏杆菌（Escherichia coli）,利用五种不同方法提取大肠埃希氏杆菌的基因组DNA .通过定性、定量分析加以比较 ,确定一套经济、有效的适用于提取大肠埃希氏杆菌基因组DNA的方法--液氮法.以该方法提取的DNA为模板,通过扩增核糖体基因区间序列（16s-23s基因间隔区）可以应用于海水中粪便污染源示踪研究.     

Faecal Coliform Bacteria in Febros River (Northwest Portugal): Temporal Variation, Correlation with Water Parameters, and Species Identification

Febros river water was sampled weekly, during 35 successive weeks, and analyzed for microbiological (total coliforms, faecal coliforms, faecal streptococci and enterococci) and chemical-physical (ammonia and temperature) parameters. All microbiological parameters were highly correlated with each other and with ammonia, suggesting that the simultaneous determination of all variables currently in use in the evaluation of the microbiological quality of waters is probably redundant, and could be simplified, and that ammonia should be tested as a sentinel parameter of the microbiological pollution load of Febros river. From the strains isolated from positive tubes of the faecal coliforms test (multiple tube fermentation technique) and retested in this assay, Escherichia coli, Klebsiella oxytoca and Klebsiella pneumoniae subsp. pneumoniae strains were positive, indicating that the faecal coliforms test is not totally specific for Escherichia coli, and can detect other bacteria. Considering that these Klebsiella spp. are not necessarily of faecal origin, it was concluded that the faecal coliforms test can overestimate true faecal pollution. From the strains isolated from positive tubes of the faecal coliforms procedure, only Escherichia coli strains were clearly positive in the β-D-glucuronidase test. All other species were negative or very weakly positive, suggesting that the assay of the β-D-glucuronidase activity is less prone to false positives than the faecal coliforms test in the quantification of Escherichia coli in environmental waters.     

大肠杆菌对赤子爱胜蚓体表超微结构的影响

以赤子爱胜蚓(Eisenia fetida)为实验动物,研究了细菌诱导(浸浴诱导和注射诱导)对蚯蚓体表超微结构的影响.结果表明:(1)用细菌浸浴4h的实验组、用细菌注射4h的实验组及对照组蚯蚓的角质层厚度分别为(0.51±0.03)μm、(0.56±0.02)μm、(0.50±0.01)μm;上角质层突起的高度分别为(0.16±0.02)μm、(0.12±0.01)μm、(0.11±0.01)μm;上角质层游离面中微绒毛的高度分别为(0.18±0.02)μm、(0.11±0.01)μm、(0.11±0.01)μm.统计分析表明,蚯蚓在抵御微生物侵袭过程中,体表角质层中的上角质层突起及上角质层游离面中的微绒毛发挥着重要作用.(2)细菌浸浴4h的实验组、用细菌注射4h实验组及对照组蚯蚓体表的表皮厚度分别为(2.63±0.15)μm、(2.23±0.06)μm、(1.97±0.15)μm;表皮中小颗粒蛋白细胞的颗粒直径分别为(0.38±0.04)μm、(0.26±0.02)μm、(0.24±0.02)μm;大颗粒粘液细胞的颗粒直径分别为(0.50±0.04)μm、(0.26±0.03)μm、(0.21±0.02)μm;网状粘液细胞的直径分别为(0.93±0.06)μm、(0.63±0.06)μm、(0.28±0.03)μm.统计分析表明,蚯蚓在抵御微生物侵袭的过程中,表皮中的腺细胞(小颗粒蛋白细胞、大颗粒粘液细胞及网状粘液细胞)发挥着重要作用.图4表2参17     

黑水虻转化厨余垃圾过程中病原菌灭活规律的研究与综合评价

黑水虻能够有效取食厨余垃圾等有机固体废弃物,其自身转化为昆虫蛋白和脂肪等生物质,将厨余垃圾转化为虫沙有机肥,处理过程是一种有机固体废弃物的资源化新方法.为评估黑水虻处理厨余垃圾的效率与安全性,本课题研究黑水虻在厨余垃圾处理过程中,对大肠杆菌O157：H7（EC）、鼠伤寒沙门氏菌（ST）及金黄色葡萄球（SA）的灭活能力,评价黑水虻体内的抑菌因子,分析3种病原菌对黑水虻生长增重及厨余转化效率的影响.经过18 d的黑水虻处理,研究发现：①第0和第6 d两次以6.4~7.1 log₁₀ CFU·g^-1的浓度向厨余垃圾中分别接种EC、ST和SA以后,EC经4~6 d处理被全部灭活、ST经3~4 d处理被全部灭活,SA经6 d处理,浓度下降到1.9~2.6 log₁₀ CFU·g^-1,但不能被全部灭活,病原菌的灭活效率呈现EC=ST>SA的趋势（p < 0.001）,且黑水虻体内无EC、ST或SA残留;②厨余垃圾降解过程中,初期和中期pH值主要呈现酸性（4.0~5.3）,对抑制EC、ST和SA起到了促进作用,后期pH值呈中性至弱碱性;③黑水虻对于EC和ST能够产生自身免疫抑制因子,且抑制活性ST>EC,但对SA无明显的免疫抑制能力,SA的灭活主要依赖于黑水虻肠道菌群的竞争性抑制作用;④黑水虻的体长、体重、预蛹率、产率,以及厨余垃圾的生物转化率和减量化率未受病原菌存在的影响,18 d后EC组、ST组和SA组的预蛹率均达到在80%以上,厨余减量化率分别达到74.0%、79.1%和78.5%,生物转化率分别达到13.0%、13.2%、19.4%.综上,黑水虻能够在彻底灭活EC、ST,99%灭活SA的同时,高效降解厨余垃圾（减量化率>74%）,并转化为虫体有机质（虫产率>10%）,是一种高效卫生的厨余垃圾资源化方法.今后的研究中,还需要加强以金葡菌为代表的抗逆性病原菌的灭活机制的探讨.     

Photocatalytic inactivation of Escherichia coli by natural sphalerite suspension: effect of spectrum, wavelength and intensity of visible light

The photocatalytic disinfection of Escherichia coli K-12 is investigated by the natural sphalerite (NS) under different spectra, wavelengths and intensities of visible light (VL) emitted by light-emitting-diode lamp (LED). The spectrum effect of VL on disinfection efficiency is studied by using white LED, fluorescent tube (FT) and xenon lamp (XE), which indicates that the “discreted peak spectrum” of FT is more effective to inactivate bacteria than “continuous spectrum” of LED and XE. Besides, the photocatalytic disinfection of bacteria is compared under different single spectrum (blue, green, yellow and red color) LEDs. The results show that the most effective wavelength ranges of VL for photocatalytic disinfection with the NS are 440-490 and 570-620 nm. Furthermore, a positive relationship is obtained between the disinfection efficiency and the VL intensity. The experiment shows that NS can completely inactivate 10⁷ cfu mL⁻¹E. coli K-12 within 8 h irradiation by white LED with the intensity of 200 mW cm⁻² at pH 8. Moreover, the destruction process of the cell wall and the cell membrane are directly observed by TEM. Finally, no bacterial colony can be detected within a 96 h regrowth test of inactivated bacteria, which reveals that the VL-photocatalytic disinfection leads to an irreversible damage to the bacterial cells.     

Inactivation and injury assessment of Escherichia coli during solar and photocatalytic disinfection in LDPE bags

Solar disinfection (SODIS) of Escherichia coli suspensions in low-density polyethylene bag reactors was investigated as a low-cost disinfection method suitable for application in developing countries. The efficiency of a range of SODIS reactor configurations was examined (single skin (SS), double skin, black-backed single skin, silver-backed single skin (SBSS) and composite-backed single skin) using E. coli suspended in model and real surface water. Titanium dioxide was added to the reactors to improve the efficiency of the SODIS process. The effect of turbidity was also assessed. In addition to viable counts, E. coli injury was characterised through spread-plate analysis using selective and non-selective media. The optimal reactor configuration was determined to be the SBSS bag (t₅₀ = 9.0 min) demonstrating the importance of UVA photons, as opposed to infrared in the SODIS disinfection mechanism. Complete inactivation (6.5-log) was achieved in the presence of turbidity (50 NTU) using the SBSS bag within 180 min simulated solar exposure. The addition of titanium dioxide (0.025 g L⁻¹) significantly enhanced E. coli inactivation in the SS reactor, with 6-log inactivation observed within 90 min simulated solar exposure. During the early stages of both SODIS and photocatalytic disinfection, injured E. coli were detected; however, irreversible injury was caused and re-growth was not observed. Experiments under solar conditions were undertaken with total inactivation (6.5-log) observed in the SS reactor within 240 min, incomplete inactivation (4-log) was observed in SODIS bottles exposed to the same solar conditions.