Bioreporter

Bioreporter
  • 文章类型: Journal Article
    自从近30年前首次引入全细胞生物报道分子(WCB)以来,他们的高灵敏度,选择性,和现场检测的适用性使它们在环境监测中具有很大的前景,医学诊断,食品安全,生物制造,和其他领域。特别是在环境领域,该技术提供了一种快速有效的方法来评估环境中污染物的生物利用度。尽管有这些优势,该技术尚未商业化。这种商业化的缺乏令人困惑,鉴于WCBs广阔的应用前景。多年来,许多研究论文主要集中在增强WCB的灵敏度和选择性上,很少关注它们更广泛的商业应用。到目前为止,目前还没有关于这个主题的批判性审查。因此,在本文中,我们批判性地回顾了WCB在过去三十年中的研究进展,评估当前系统的性能和局限性,以了解商业部署的障碍。通过识别这些障碍,本文为研究人员和行业利益相关者提供了对阻碍市场进入的挑战的更深入的见解,并激发了克服这些障碍的进一步研究,从而促进WCB的商业化,作为一种有前途的环境监测技术。
    Since the initial introduction of whole-cell bioreporters (WCBs) nearly 30 years ago, their high sensitivity, selectivity, and suitability for on-site detection have rendered them highly promising for environmental monitoring, medical diagnosis, food safety, biomanufacturing, and other fields. Especially in the environmental field, the technology provides a fast and efficient way to assess the bioavailability of pollutants in the environment. Despite these advantages, the technology has not been commercialized. This lack of commercialization is confusing, given the broad application prospects of WCBs. Over the years, numerous research papers have focused primarily on enhancing the sensitivity and selectivity of WCBs, with little attention paid to their wider commercial applications. So far, there is no a critical review has been published yet on this topic. Therefore, in this article we critically reviewed the research progress of WCBs over the past three decades, assessing the performance and limitations of current systems to understand the barriers to commercial deployment. By identifying these obstacles, this article provided researchers and industry stakeholders with deeper insights into the challenges hindering market entry and inspire further research toward overcoming these barriers, thereby facilitating the commercialization of WCBs as a promising technology for environmental monitoring.
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  • 文章类型: Journal Article
    这里,我们证明了产生表面活性剂的假单胞菌对Pantoeaeucalypti299R的有益作用。我们在日益复杂的环境中进行了一系列实验。P.桉树树299R(Pe299R),和假单胞菌。FF1(Pff1)或Pe299R和表面活性剂生产缺陷的假单胞菌属。FF1::将Δ内脏B(Pff1Δ内脏B)共同接种在肉汤中,在成群的琼脂平板上,和植物。在肉汤中,在Pff1或Pff1Δ内脏B存在下生长时,Pe299R的生长动力学没有差异。相比之下,在成群的琼脂平板上,与使用Pff1Δ内脏B或单一培养生长的Pe299R相比,Pe299R能够与Pff1共同聚集,从而导致Pe299R生物量显着增加。最后在植物中,并使用单细胞生物报告子进行生殖成功(CUSPER),我们发现Pff1对共同接种的Pe299R亚群具有时间上明显的有益作用,而在Pff1Δ内脏B存在下不会发生。我们在蜂群琼脂上的PE299R上测试了三种其他产生表面活性剂的假单胞菌及其各自的表面活性剂敲除突变体,显示出相似的结果。这导致我们提出了一种模型,用于在叶片定殖过程中产生表面活性剂的积极作用。我们的结果表明,在叶片定殖过程中,共运动性可能是常见的,并且为表面活性剂已经具有多种作用的又一个方面。
    Here, we demonstrate the beneficial effect of surfactant-producing pseudomonads on Pantoea eucalypti 299R. We conducted a series of experiments in environments of increasing complexity. P. eucalypti 299R (Pe299R), and Pseudomonas sp. FF1 (Pff1) or Pe299R and surfactant-production deficient Pseudomonas sp. FF1::ΔviscB (Pff1ΔviscB) were co-inoculated in broth, on swarming agar plates, and on plants. In broth, there were no differences in the growth dynamics of Pe299R when growing in the presence of Pff1 or Pff1ΔviscB. By contrast, on swarming agar plates, Pe299R was able to co-swarm with Pff1 which led to a significant increase in Pe299R biomass compared to Pe299R growing with Pff1ΔviscB or in monoculture. Finally in planta, and using the single-cell bioreporter for reproductive success (CUSPER), we found a temporally distinct beneficial effect of Pff1 on co-inoculated Pe299R subpopulations that did not occur in the presence of Pff1ΔviscB. We tested three additional surfactant-producing pseudomonads and their respective surfactant knockout mutants on PE299R on swarming agar showing similar results. This led us to propose a model for the positive effect of surfactant production during leaf colonization. Our results indicate that co-motility might be common during leaf colonization and adds yet another facet to the already manyfold roles of surfactants.
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  • 文章类型: Journal Article
    豆类结瘤需要根瘤菌检测根际中的类黄酮,以激活它们产生的Nod因子反信号。在这里,我们研究了与苜蓿结瘤有关的类黄酮。我们对五种类黄酮-O-甲基转移酶(OMT)进行了生化表征,并使用了基于lux的nod基因报告基因来研究中华根瘤菌NodD1对各种类黄酮的反应。我们发现查尔酮-OMT1(ChOMT1)和ChOMT3,而不是OMT2,4和5,能够产生4,4'-二羟基-2'-甲氧基查尔酮(DHMC)。生物记者对DHMC反应最强烈,而对大豆结瘤重要的异黄酮(Glycinemax)则没有活性。突变体分析显示ChOMT1的缺失强烈降低了DHMC水平。此外,chomt1和omt2在其根际显示出大大降低的生物报告发光。此外,ChOMT1和ChOMT3的丢失减少了结瘤,OMT2的进一步丢失加强了这种表型。我们得出的结论是:ChOMT1的丢失大大降低了根DHMC的水平;ChOMT1或OMT2对于根际节点基因的激活很重要;ChOMT1/3和OMT2促进结瘤。我们的发现表明,与大豆相比,在M.truncatula中用于结瘤的类黄酮具有一定程度的排他性,支持类黄酮在根瘤菌寄主范围中的作用。
    Legume nodulation requires the detection of flavonoids in the rhizosphere by rhizobia to activate their production of Nod factor countersignals. Here we investigated the flavonoids involved in nodulation of Medicago truncatula. We biochemically characterized five flavonoid-O-methyltransferases (OMTs) and a lux-based nod gene reporter was used to investigate the response of Sinorhizobium medicae NodD1 to various flavonoids. We found that chalcone-OMT 1 (ChOMT1) and ChOMT3, but not OMT2, 4, and 5, were able to produce 4,4\'-dihydroxy-2\'-methoxychalcone (DHMC). The bioreporter responded most strongly to DHMC, while isoflavones important for nodulation of soybean (Glycine max) showed no activity. Mutant analysis revealed that loss of ChOMT1 strongly reduced DHMC levels. Furthermore, chomt1 and omt2 showed strongly reduced bioreporter luminescence in their rhizospheres. In addition, loss of both ChOMT1 and ChOMT3 reduced nodulation, and this phenotype was strengthened by the further loss of OMT2. We conclude that: the loss of ChOMT1 greatly reduces root DHMC levels; ChOMT1 or OMT2 are important for nod gene activation in the rhizosphere; and ChOMT1/3 and OMT2 promote nodulation. Our findings suggest a degree of exclusivity in the flavonoids used for nodulation in M. truncatula compared to soybean, supporting a role for flavonoids in rhizobial host range.
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  • 文章类型: Journal Article
    自1962年发现荧光蛋白以来,它们已经彻底改变了科学。他们使成像实验能够破译蛋白质的功能,细胞,和有机体,以及基因调控。绿色荧光蛋白及其衍生物现在是细胞生物学的标准工具,免疫学,分子生物学,和世界各地的微生物实验室。这些蛋白质的一个共同特征是它们依赖双氧(O2)的成熟允许荧光,这就排除了它们在缺氧环境中的使用。在这项工作中,我们报告了编码不依赖O2的KOFP-7蛋白的遗传回路的发展和细胞表征,一种黄素结合荧光蛋白。我们在种群和单细胞水平上优化了高细菌荧光的遗传电路,在宿主范围不同的各种质粒中实现该电路,并在有氧和厌氧条件下定量它们的荧光。最后,我们表明,基于KOFP-7的结构可用于产生重氮弧菌的荧光细胞,兼性厌氧菌,证明了遗传回路对各种厌氧细菌的有用性。因此,这些遗传回路可以随意修改,解决基础和应用研究问题,开辟一条高速公路,照亮晦涩的无氧世界。重要荧光蛋白已经使用了几十年,并在广泛的领域中获得了生物学的重大发现,并用于环境和临床环境。绿色荧光蛋白(GFP)及其所有衍生物都有一个共同的特征:它们依赖于双氧(O2)的存在来实现蛋白质的成熟和荧光。这种依赖性排除了它们在缺氧环境中的使用。这里,我们构建了一系列遗传回路,允许产生KOFP-7,一种与O2无关的黄素结合荧光蛋白。我们证明了产生KOFP-7的大肠杆菌细胞是荧光的,在群体和单细胞水平。重要的是,我们证明了,与产生GFP的细胞不同,产生KOFP-7的细胞在缺氧时发出荧光。最后,我们证明了兼性厌氧菌重氮弧菌NS1,当产生KOFP-7时,在不存在O2的情况下是荧光的。总之,允许O2无关荧光的新遗传电路的开发将为研究厌氧过程开辟新的视角。
    Fluorescent proteins have revolutionized science since their discovery in 1962. They have enabled imaging experiments to decipher the function of proteins, cells, and organisms, as well as gene regulation. Green fluorescent protein and all its derivatives are now standard tools in cell biology, immunology, molecular biology, and microbiology laboratories around the world. A common feature of these proteins is their dioxygen (O2)-dependent maturation allowing fluorescence, which precludes their use in anoxic contexts. In this work, we report the development and in cellulo characterization of genetic circuits encoding the O2-independent KOFP-7 protein, a flavin-binding fluorescent protein. We have optimized the genetic circuit for high bacterial fluorescence at population and single-cell level, implemented this circuit in various plasmids differing in host range, and quantified their fluorescence under both aerobic and anaerobic conditions. Finally, we showed that KOFP-7-based constructions can be used to produce fluorescing cells of Vibrio diazotrophicus, a facultative anaerobe, demonstrating the usefulness of the genetic circuits for various anaerobic bacteria. These genetic circuits can thus be modified at will, both to solve basic and applied research questions, opening a highway to shed light on the obscure anaerobic world.IMPORTANCEFluorescent proteins are used for decades, and have allowed major discoveries in biology in a wide variety of fields, and are used in environmental as well as clinical contexts. Green fluorescent protein (GFP) and all its derivatives share a common feature: they rely on the presence of dioxygen (O2) for protein maturation and fluorescence. This dependency precludes their use in anoxic environments. Here, we constructed a series of genetic circuits allowing production of KOFP-7, an O2-independant flavin-binding fluorescent protein. We demonstrated that Escherichia coli cells producing KOFP-7 are fluorescent, both at the population and single-cell levels. Importantly, we showed that, unlike cells producing GFP, cells producing KOFP-7 are fluorescent in anoxia. Finally, we demonstrated that Vibrio diazotrophicus NS1, a facultative anaerobe, is fluorescent in the absence of O2 when KOFP-7 is produced. Altogether, the development of new genetic circuits allowing O2-independent fluorescence will open new perspective to study anaerobic processes.
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  • 文章类型: Journal Article
    在这项研究中,草甘膦(GLY)的干扰作用,氨甲基膦酸(AMPA),在斑马鱼急性暴露于环境相关浓度的120小时内,首次报道了三种基于草甘膦的除草剂(GBHs)以非浓度依赖性方式对卵黄发生的影响。GBHs在全球范围内普遍用于杂草控制管理。由于其广泛应用,它们经常发生在水生生态系统中,并可能影响各种生物。活性物质GLY及其主要副产品,AMPA,是研究最彻底的化学物质;然而,GBHs复杂配方的不利影响与不同和未知含量的共同配方仍未充分研究。这项研究集中在胚胎毒性,亚致死畸形,和GLY的雌激素效力,AMPA,和四种常用的GBHs在斑马鱼胚胎上使用野生型和雌激素敏感,转基因斑马鱼品系(Tg(vtg1:mCherry))。经过120小时的博览会,AMPA没有引起急性毒性,GLY的LC50为160mg/L。GBHs毒性更大,LC50值范围为31至111GLY活性当量(a.e.)mg/L。暴露于0.35-2.8mg/LGBH会导致亚致死性异常:典型症状是下颌的结构变形和嗅觉区域的异常。治疗组的畸形率为10-30%。在体内,通过非侵入性显微镜方法检测到胚胎肝脏中荧光表达的vtg1mCherry蛋白,表明通过GLY产生卵黄蛋白原的雌激素作用,AMPA,GBHs。为了确认体内发现,进行RT-qPCR方法以确定雌激素相关的vtg1mRNA的水平。暴露于GLY120小时后,AMPA,和三个浓度为0.35mg/L的GBHs,vtg1基因表达显著上调。我们的研究结果强调了短期GLY和GBH暴露可能导致斑马鱼胚胎发育畸形并破坏激素平衡的风险。
    In this study, the disrupting effects of glyphosate (GLY), aminomethylphosphonic acid (AMPA), and three glyphosate-based herbicides (GBHs) on vitellogenesis in a non-concentration-dependent manner are reported for the first time in 120 h of acute exposure of zebrafish at environmentally relevant concentrations. GBHs are commonly used worldwide in weed control management. Due to their extensive application, they frequently occur in aquatic ecosystems and may affect various organisms. The active substance GLY and its major by-product, AMPA, are the most thoroughly studied chemicals; however, the adverse effects of the complex formulas of GBHs with diverse and unknown content of co-formulants are still not sufficiently researched. This study focused on the embryotoxicity, sublethal malformations, and estrogenic potency of GLY, AMPA, and four commonly used GBHs on zebrafish embryos using a wild type and an estrogen-sensitive, transgenic zebrafish line (Tg(vtg1:mCherry)). After 120 h of exposition, AMPA did not cause acute toxicity, while the LC50 of GLY was 160 mg/L. The GBHs were more toxic with LC50 values ranging from 31 to 111 GLY active equivalent (a.e.) mg/L. Exposure to 0.35-2.8 mg/L GBHs led to sublethal abnormalities: typical symptoms were structural deformation of the lower jaw and anomalies in the olfactory region. Deformity rates were 10-30% in the treated groups. In vivo, fluorescently expressed vtg1 mCherry protein in embryonic liver was detected by a non-invasive microscopic method indicating estrogenic action through vitellogenin production by GLY, AMPA, and GBHs. To confirm the in vivo findings, RT-qPCR method was performed to determine the levels of the estrogenicity-related vtg1 mRNA. After 120 h of exposure to GLY, AMPA, and three GBHs at a concentration of 0.35 mg/L, the expression of vtg1 gene was significantly up-regulated. Our results highlight the risk that short-term GLY and GBH exposure can cause developmental malformations and disrupt the hormonal balance in zebrafish embryos.
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  • 文章类型: Journal Article
    水体富营养化,由于工业和农业废水的排放,导致生态退化。然而,到目前为止,如何评估和管理水污染的风险,尤其是氮污染,仍然是一个特别值得注意的问题。硝酸盐,最重要的含氮化合物,已成为制约全氮管理的瓶颈。监测硝酸盐污染的生物记者的发展有助于估计水质,尤其是营养的可用性。在这项研究中,我们从基于不同宿主构建的40种蓝藻衍生物中获得了9种生物报告基因,副本编号,和传感元件,并评估了生物报道分子的性能。结果表明,单细胞集胞藻对硝酸盐的敏感性高于丝状鱼,在大多数生物报告基因中,报道基因luxABCDE的反应比sfgfp快,相对中等拷贝的质粒提高了传感元件的性能。九个生物报告基因在生物利用硝酸盐检测中表现良好,其中AD-AS-X和AR-NI-X,通过硝酸盐填充活化,具有最短的响应时间(2小时)和最宽的响应范围(20-800μM),分别。此外,SR-GLN-SG,硝酸盐缺乏激活,表现出最佳的线性响应(R2=0.998)。参数优化后,指数生长期生物记者,培养温度为30°C,将200μL的样品体积确定为最佳监测条件。我们发现常见的水污染物(铜,镉,和磷)对生物记者的性能没有影响,表明生物记者的稳定性。9名生物记者中有6名,尤其是SR-NB-X,在检测废水样品中的生物可利用硝酸盐方面非常有效。该研究为开发更多蓝藻生物分子及其在硝酸盐检测中的实际应用提供了有价值的参考。
    The water eutrophication, resulting from the discharge of industrial and agricultural wastewater, leads to ecological degradation. However, to date, how to assess and manage the risks of water pollution, especially nitrogen pollution, remains a particularly noteworthy issue. Nitrate, the most important nitrogen compound, has become a bottleneck restricting total nitrogen management. The development of bioreporters monitoring nitrate pollution contributes to the estimation of water quality, especially the availability of nutrients. In this study, we obtained 9 bioreporters from 40 cyanobacterial derivatives which were constructed based on different hosts, copy numbers, and sensing elements and evaluated the performance of bioreporters. The results showed that single-celled Synechocystis was more sensitive to nitrate than filamentous Anabaena, that the reporter gene luxABCDE responded faster than sfgfp in most bioreporters, and that relatively medium-copy plasmid improved the performance of sensing elements. Nine bioreporters performed well in bioavailable nitrate detection, of which AD-AS-X and AR-NI-X, activated by nitrate repletion, had the shortest response time (2 h) and the widest response range (20-800 μM), respectively. Moreover, SR-GLN-SG, activated by nitrate deficiency, exhibited the best linear response (R2 = 0.998). After parameter optimization, exponential growth phase bioreporters, culture temperature of 30 °C, sample volume of 200 μL were determined as optimal monitoring conditions. We found that common water contaminants (copper, cadmium, and phosphorus) had no impact on the performance of bioreporters, indicating the stability of bioreporters. Six out of 9 bioreporters, especially the SR-NB-X, were highly effective in detecting the bioavailable nitrate in wastewater sample. This study provides valuable references for developing more cyanobacterial bioreporters and their practical application in nitrate detection.
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  • 文章类型: Journal Article
    这项研究的目的是确定最佳条件,以提高金和银纳米颗粒抗玉米赤霉烯酮抗体缀合物的稳定性,用于侧流免疫层析测定(LFIA)。Turkevich-Frens方法用于合成金纳米粒子(AuNPs),直径在10到110纳米之间。使用硼氢化钠作为还原剂合成尺寸分布为2.5至100nm的银纳米颗粒(AgNP)。AuNP和AgNP聚集的开始发生在150mM和80mMNaCl浓度,分别。在1.2mM的K2CO3浓度下获得稳定的Au和Ag纳米颗粒-抗体缀合物,对应于约7的pH值。最后,在抗体浓度为4和6µg/mL时,Au和Ag纳米颗粒与抗玉米赤霉烯酮抗体的缀合程度最高.缀合条件的优化可以有助于纳米颗粒及其抗体缀合物的更好稳定性,并且可以改善生物传感侧流装置中生物报告分子结果的再现性。
    The aim of this research is to define optimal conditions to improve the stability of gold and silver nanoparticles\' anti-zearalenone antibody conjugates for their utilisation in lateral flow immunochromatographic assay (LFIA). The Turkevich-Frens method was used to synthesise gold nanoparticles (AuNPs), which were between 10 and 110 nm in diameter. Silver nanoparticles (AgNPs) with a size distribution of 2.5 to 100 nm were synthesised using sodium borohydride as a reducing agent. The onset of AuNP and AgNP aggregation occurred at 150 mM and 80 mM NaCl concentrations, respectively. Stable Au and Ag nanoparticle-antibody conjugates were achieved at 1.2 mM of K2CO3 concentration, which corresponds to the pH value of ≈7. Lastly, the highest degree of conjugation between Au and Ag nanoparticles and anti-zearalenone antibodies was at 4 and 6 µg/mL of antibody concentrations. The optimisation of the conjugation conditions can contribute to better stability of nanoparticles and their antibody conjugate and can improve the reproducibility of results of bioreporter molecules in biosensing lateral flow devices.
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  • 文章类型: Journal Article
    Introduction.肺炎克雷伯菌是全球公共卫生的主要威胁。它是包括尿路感染在内的多种疾病表现的病原体,败血症,肝脓肿,伤口感染和呼吸道感染。肺炎克雷伯菌导致社区和医院获得性肺炎,这是一种与高死亡率相关的毁灭性疾病。假说。人们越来越关注多药耐药肺炎克雷伯菌菌株的出现,使目前可用的治疗方法复杂化;因此,迫切需要开发新的抗菌剂。瞄准.肺炎克雷伯菌在小鼠中引起急性呼吸道疾病,并且在目前的工作中,我们研究了对生物发光克雷伯菌进行非侵入性监测以监测治疗效果的能力。方法论。我们设计了肺炎克雷伯菌的生物发光报告菌株,以监测抗生素在鼠呼吸道疾病模型中的影响。结果。我们证明了生物发光与宿主组织中的细菌数量相关,从而可以对体内细菌复制进行非侵入性计数。光的产生与细菌的生存能力直接相关,这种新型生物发光肺炎克雷伯菌菌株能够监测美罗培南治疗阻止肺部细菌增殖的功效。结论。非侵入性生物发光成像的使用改善了临床前动物模型测试,以更早和更高的灵敏度检测研究结果。
    Introduction. Klebsiella pneumoniae is a major threat to public health worldwide. It is the causative agent for multiple disease presentations including urinary tract infection, septicemia, liver abscess, wound infection and respiratory tract infection. K. pneumoniae causes community- and hospital-acquired pneumonia, which is a devastating disease associated with high mortality rates.Hypothesis. There is a growing concern about the emergence of multidrug-resistant K. pneumoniae strains complicating the treatment with the current available therapeutics; therefore, there is an urgent need for the development of new antimicrobial agents.Aim. K. pneumoniae causes an acute respiratory disease in mice and in the current work we investigated the capability to perform non-invasive monitoring of bioluminescent Klebsiella to monitor therapeutic efficacy.Methodology. We engineered a bioluminescence reporter strain of K. pneumoniae to monitor the impact of antibiotics in a murine respiratory disease model.Results. We demonstrate that bioluminescence correlates with bacterial numbers in host tissues allowing for a non-invasive enumeration of bacterial replication in vivo. Light production is directly linked to bacterial viability, and this novel bioluminescent K. pneumoniae strain enabled monitoring of the efficacy of meropenem therapy in arresting bacterial proliferation in the lung.Conclusion. The use of non-invasive bioluminescent imaging improves preclinical animal model testing to detect study outcome earlier and with higher sensitivity.
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  • 文章类型: Journal Article
    三年来,大量的制造污染物,如塑料,由于COVID-19,抗生素和消毒剂已被释放到环境中。这些污染物在环境中的积累加剧了对土壤系统的破坏。然而,自从疫情爆发以来,研究人员和公众关注的焦点一直集中在人类健康上。值得注意的是,与土壤污染和CIVID-19结合进行的研究仅占所有COVID-19研究的4%。为了提高研究人员和公众对COVID-19土壤污染严重性的认识,我们提出了“大流行COVOD-19结束但土壤污染增加”的观点,并推荐了一种基于全细胞生物传感器的新方法来评估COVID-19衍生污染物的环境风险。该方法有望为大流行产生的污染物影响的土壤的环境风险评估提供新的方法。
    For three years, a large amount of manufactured pollutants such as plastics, antibiotics and disinfectants has been released into the environment due to COVID-19. The accumulation of these pollutants in the environment has exacerbated the damage to the soil system. However, since the epidemic outbreak, the focus of researchers and public attention has consistently been on human health. It is noteworthy that studies conducted in conjunction with soil pollution and COVID-19 represent only 4 % of all COVID-19 studies. In order to enhance researchers\' and the public awareness of the seriousness on the COVID-19 derived soil pollution, we propose the viewpoint that \"pandemic COVID-19 ends but soil pollution increases\" and recommend a whole-cell biosensor based new method to assess the environmental risk of COVID-19 derived pollutants. This approach is expected to provide a new way for environmental risk assessment of soils affected by contaminants produced from the pandemic.
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  • 文章类型: Journal Article
    溢油事故频发,威胁着生态系统和人类健康。固相微萃取允许从环境基质中直接提取烷烃以提高检测极限,但无法现场测量烷烃。通过将烷烃趋化性不动杆菌生物报告基因ADPWH_alk固定在琼脂糖凝胶中,开发了一种生物相微萃取和生物传感(BPME-BS)装置,借助光电倍增管实现在线烷烃定量。BPME-BS装置具有高富集因子(平均7.07)和令人满意的烷烃检测限(0.075mg/L)。定量范围为0.1-100mg/L,与气相色谱火焰电离检测器相当,比没有固定的生物报告更好。BPME-BS器件中的ADPWH_alk细胞在广泛的环境条件下保持良好的灵敏度,包括pH(4.0-9.0),温度(20-40°C),和盐度(0.0-3.0%),其反应在4℃下30天内保持稳定。在7天的连续测量中,BPME-BS设备成功地可视化了烷烃的动态浓度,为期7天的现场测试成功捕获了一起漏油事件,帮助进行来源分配和现场执法。我们的工作证明,BPME-BS设备是在线测量烷烃的强大工具,显示出快速检测和快速响应现场和现场漏油的巨大潜力。
    Oil spill incidents occur frequently and threaten ecosystems and human health. Solid-phase microextraction allows direct alkane extraction from environmental matrices to improve the limit of detection but is unable to measure alkanes on site. A biological-phase microextraction and biosensing (BPME-BS) device was developed by immobilising an alkane chemotactic Acinetobacter bioreporter ADPWH_alk in agarose gel to achieve online alkane quantification with the aid of a photomultiplier. The BPME-BS device had a high enrichment factor (average 7.07) and a satisfactory limit of detection (0.075 mg/L) for alkanes. The quantification range was 0.1-100 mg/L, comparable to a gas chromatography flame ionisation detector and better than a bioreporter without immobilisation. ADPWH_alk cells in the BPME-BS device maintained good sensitivity under a wide range of environmental conditions, including pH (4.0-9.0), temperature (20-40 °C), and salinity (0.0-3.0%), and its response remained stable within 30 days at 4 °C. In a 7-day continual measurement, the BPME-BS device successfully visualised the dynamic concentration of alkanes, and a 7-day field test successfully captured an oil spill event, helping in source apportionment and on-scene law enforcement. Our work proved that the BPME-BS device is a powerful tool for online alkane measurement, showing substantial potential for fast detection and rapid response to oil spills on site and in situ.
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