Bacteria and the Biodegradation of Chemicals Achieved Naturally,by Combination,or by Construction

The natural potential of bacteria for the biological degradation of synthetic compounds is greater than is commonly supposed and extends to many heteroarenes and even some chloroarenes. An increase in the number of substituents on the aromatic ring or a certain substitution pattern is what confers xenobiotic character to a compound. In addition, when enzymes with low substrate specificity encounter foreign compounds with random variations, products with very strong xenobiotic character often result. In this case, changing the conditions or introducing a cooperation between several different types of bacteria can be used to degrade these compounds. Finally, mineralization, the complete breakdown of organic substances into carbon dioxide and inorganic salts, of xenobiotics previously regarded as persistent can be achieved by taking advantage of natural or induced gene transfer to construct hybrid degradative pathways. After an introduction to the world of bacteria and their place in Nature, we will describe their natural potential for biodegradation with reference to aliphatic and aromatic hydrocarbons. The discussion will then turn to the types of the substituents that confer xenobiotic properties to compounds and how these compounds are degraded despite their xenobiotic character.     

On-line extraction of metal ions using liquid-liquid segmentation and a membrane type phase separator for fluorescence detection

Summary A liquid segmented post-column reaction system has been used to extract metal ions from an aqueous eluent into an organic solvent for fluorescence detection. The metals Zr(IV), Ga(III), Sc(III), Y(III), In(III), Al(III), La(III), Zn(II), Cd(II), Ca(II) and Mg(II) have been isocratically separated on a C₁₈ column by virtue of the secondary chemical equilibrium established by an eluent containing n-octanesulfonate, tartaric acid, and hydroxyisobutyric acid. The chelating reagent 8-hydroxyquinoline dissolved in methylisobutyl ketone (MIBK) was used to extract the metals and a membrane type phase separator was effective at separating the phases and directing the organic stream to the detector. The response for this detection approach was linear for metal ion concentrations spanning the range of the detector, and detection limits for most metals were low parts-per-billion (ppb). Band broadening for the extraction system was examined and compared to a direct post-column reaction using oxine dissolved in acetone.     

The effect of frequency and power density on the ultrasonically-enhanced killing of biofilm-sequestered Escherichia coli

Infection on implanted medical devices is a critical concern because the bacteria are recalcitrant to antibiotic therapy; currently the only way to eliminate the infection is to remove the device. We have found that low-frequency ultrasound renders bacteria more susceptible to antibiotics. The effect of low-intensity ultrasound on the enhancement of antibiotic action against biofilm bacteria was measured by subjecting thick E. coli biofilms for 2 h at 37°C to one of four conditions: (1) incubation in nutrient broth; (2) incubation in nutrient broth with antibiotic; (3) ultrasonication in nutrient broth without antibiotic; and (4) ultrasonication in nutrient broth with antibiotic. Two frequencies (70 and 500 kHz) and several ultrasonic intensities were examined, ranging from 2 to 200 mW/cm². It was determined that low-intensity ultrasound significantly enhanced killing of biofilm E. coli by gentamicin. This enhancement increased with increasing ultrasonic intensity and decreased with increasing frequency. A mathematical model of ultrasonically-enhanced transport in cylindrical pores and channels shows that gentamicin transport increases with ultrasonic intensity and decreases with increasing frequency. However, the magnitude of increased transport is so small that it is difficult to attribute enhanced killing of bacteria to enhanced antibiotic transport through the pores and channels of the biofilm; therefore, other mechanisms must play a role. The use of low-intensity ultrasound in conjunction with antibiotic treatment may prove to be a viable clinical method of eliminating biofilm infections from the surfaces of implanted medical devices.     

不同金属/适配体双功能复合磁性纳米材料的制备及其对外泌体的富集性能

外泌体作为一种细胞外囊泡,其内容物可以反映亲代细胞的重要信息,而自身也具有独特的结构,能够执行特征的生物学功能。基于外泌体的表面化学和生物学特征,制备了不同类型的金属/适配体(Apt)双功能复合磁性纳米材料,并将其应用于外泌体的富集纯化。将适配体和外泌体表面目标膜蛋白的特异性结合性能与以钛、锆为代表的金属氧化物和外泌体磷脂双层膜的特异性亲和作用结合,可极大地提高分离材料对外泌体的分离选择性和富集容量。分别以Fe₃O₄@Zr-MOFs、Fe₃O₄@Zr-Ti-MOFs和Fe₃O₄@TiO₂等金属有机框架(MOFs)/金属氧化物磁性纳米材料为基底,制备对应的双功能MOFs/金属氧化物-适配体复合磁性纳米材料Fe₃O₄@Zr-MOFs-Apt、Fe₃O₄@Zr-Ti-MOFs-Apt和Fe₃O₄@TiO₂-Apt,并进一步对不同材料的外泌体富集性能加以评价。以超速离心法提取的模型外泌体以及尿液为样品,对修饰相同质量适配体和不同含量金属氧化物的双功能材料的富集性能加以对比。将3种双功能磁性纳米材料应用于尿液外泌体的富集,得到的外泌体裂解后经质谱鉴定,分别得到233、343和832个外泌体蛋白。这一结果也表明双功能磁性纳米材料可以充分结合核酸适配体亲和的高选择性和金属氧化物的高富集容量优势,对于复杂生物样品中外泌体的快速、高效分离纯化具有潜在的应用价值,而针对材料制备和分离纯化方法的设计也为新型外泌体富集材料的设计提供了一条可行的新思路。     

Halogenometalates with N-containing organic cations structural chemistry and thermal behavior

The crystal structures, spectroscopic characterization, and thermal behavior of fluoro, chloro, and bromo complexes preferably of aluminium, iron and manganese formed with organic cations are reviewed. Compared with the structural chemistry of complexes with alkali metal cations, the use of quarternary ammonium or protonated N-containing base cations considerably extends the structural variety of halogenocomplexes and, in certain cases, allows the access to new modifications of metal fluorides if a suitable precursor compound is thermally decomposed.

For most of the described new compounds, the formation of the final structure is governed by the anionic part. Isolated anions [MF_{6 − n}(H₂O)_n]^{(3 − n)−} connected via hydrogen bridges are the predominant structure motifs. However, multi-nuclear units as well as chain structures can be prepared from aqueous HX solutions, too. Due to the hydrogen atoms provided by the organic cations, additional hydrogen bonds stabilize various novel structures. Some special structural features will be presented, e.g. the discrete tetrafluoroaluminate complex as well as pentacoordinated aluminium and iron(II).     

Effects of slight variations in nutrient loadings on pore plugging in soil columns

The high nutrient concentrations that would exist near the nutrient injection well during the application of cometabolicin situ bioremediation may lead to the development of significant quantities of biomass at this point in the subsurface. This biomass can decrease the porosity of the soil to such an extent that nutrient injection is no longer possible. In this work, experiments were conducted using a porous media biofilm reactor, operated under constant substrate loading conditions, such that the pressure drop across the reactor was allowed to increase to maintain a constant volumetric flow rate through the reactor. Results suggest that biomass production, and hence biofilm thickness, near the injection feed port is highly sensitive to substrate loading. In addition, these variations in biofilm thickness produce dramatic differences in the pressure drop that is attained across the reactor. Use of the Kozeny-Carman equation can be used to predict that once a critical depth has been exceeded, the pressure drop across the bed will increase exponentially within biofilm depth. This result means that pressure is not a reliable indicator of the onset of pore plugging.     

Application of an electrochemical membrane reactor to the thermochemical water splitting IS process for hydrogen production

The Bunsen reaction (SO₂ + I₂ + 2H₂O = H₂SO₄ + 2HI) in the thermochemical IS process to produce hydrogen was successfully employed using an electrochemical membrane reactor. H₂SO₄ and HI were concentrated in the anode side and the cathode side of the reactor, respectively. I₂ is the dominant bulk of the recycling chemicals in this process, and I₂ concentration at the outlet of the reactor was reduced ca. 93% by using this technique. The electric energy consumption for the reaction was about 50% smaller by reducing the concentration of I₂ indicating that the IS process can be operate efficiently at low I₂ concentration. The reaction was carried out for 4 h, and the HI concentration was increased by 26%. This amount was the same within 10% as the values calculated from the total loaded electricity. In order to decrease the overpotential at the anode side, small amount of HI was added to the anode side solution. The total voltage was reduced by 0.03 V by the addition of HI.     

Untargeted screening of unknown xenobiotics and potential toxins in plasma of poisoned patients using high-resolution mass spectrometry: Generation of xenobiotic fingerprint using background subtraction

A novel analytical workflow was developed and applied for the detection and identification of unknown xenobiotics in biological samples. High-resolution mass spectrometry (HRMS)-based data-independent MS^E acquisition was employed to record full scan MS and fragment spectral datasets of test and control samples. Then, an untargeted data-mining technique, background subtraction, was utilized to find xenobiotics present only in test samples. Structural elucidation of the detected xenobiotics was accomplished by database search, spectral interpretation, and/or comparison with reference standards. Application of the workflow to analysis of unknown xenobiotics in plasma samples collected from four poisoned patients led to generation of xenobiotic profiles, which were regarded as xenobiotic fingerprints of the individual samples. Among 19 xenobiotics detected, 11 xenobiotics existed in a majority of the patients' plasma samples, thus were considered as potential toxins. The follow-up database search led to the tentative identification of azithromycin (X5), α-chaconine (X9) and penfluridol (X12). The identity of X12 was further confirmed with its reference standard. In addition, one xenobiotic component (Y5) was tentatively identified as a penfluridol metabolite. The remaining unidentified xenobiotics listed in the xenobiotic fingerprints can be further characterized or identified in retrospective analyses after their spectral data and/or reference compounds are available. This HRMS-based workflow may have broad applications in the detection and identification of unknown xenobiotics in individual biological samples, such as forensic and toxicological analysis and sport enhancement drug screening.     

Gram-negative bacteria viable in ultrapure water: identification of bacteria isolated from ultrapure water and effect of temperature on their behavior

This study describes the effect of temperature on the behavior of bacteria viable in ultrapure water and the contamination of ultrapure water by bacteria. Three species of bacteria were isolated from ultrapure water (total organic carbon, 60 ppb and 5 ppb; effluent resistivity > 18 MΩ cm at 25°C) and identified by morphological and physiological characteristics. The three isolates were incubated in water for injection and PYG broth to check the growth profile at various temperatures. In PYG broth, temperature influenced the behavior of bacteria directly; however, it did not in water for injection. By checking both viable and non-viable bacterial numbers and endotoxin concentration in pure water, the water was found to be contaminated with non-viable bacteria and newly generated endotoxins besides viable bacteria. A column treatment, a mixed bed of fully regenerated strong acid cation exchange resin (SACER) and strong base anion exchange resin (SBAER), was used to remove bacteria from pure water. Bacteria could not grow on the surfaces of ion exchange resins in the mixed bed. The removal of bacteria was more effective as pure water was circulated through the mixed bed more rapidly.     

A Comparative Toxicity Study of TiO2 Nanoparticles in Suspension and Adherent Culture Under the Dark Condition

The present study focused on the different acute toxicity of TiO₂ nanoparticles(TiO₂ NPs) towards the bacteria in suspension culture and adherent culture under the dark conditions. The study investigated the bacteria toxicity with TiO₂ NPs at different concentrations(1—2000 mg/L), sizes(10 nm, 35 nm) and specific surface areas in unit volume solution(0—224 m²/L) characterized by the cell viability, extracellular polymeric substances(EPS) release and biofilm formation. The bacteria in adherent culture was found to be more resistant against the toxicity of TiO₂ NPs compared to that in suspension culture. An NP dose and surface area dependent(rather than the size) bacterial viability was observed in suspension culture, specifically the surface area positively correlated with the toxicity of TiO₂ NPs. The size of TiO₂ NPs, however, played a more critical role in toxicity of TiO₂ NPs in adherent culture. Therefore, the surface area dependent toxicity of TiO₂ NPs is a comprehensive parameter describing the dose and size dependent toxicity of TiO₂ NPs. The electron microscopic(SEM, TEM, EDX) observations suggested the EPS release and biofilm formation, during aggregation of TiO₂ NPs on the bacteria after 12 h cultivation in adherent culture under the dark condition. A possible toxic mechanism could be that “effective surface areas” that directly contact with the bacterial membrane greatly contributed to the toxicity of TiO₂ NPs in both suspension culture and adherent culture. Therefore, as for the possible resistance mechanism, EPS secretion and subsequent biofilm formation may protect the bacteria against the toxicity of TiO₂ NPs.     

Acute Toxicity Assay Based on Electrochemical Method for Detection of Antibiotic Residues in Water

In this work, the bacteria suspended in solution or immobilized in the biofilm were employed to study whether bacterial electrochemistry can reflect the impacts of the antibiotic residues in water on microorganisms. In the sensing system with immobilized bacteria, the tested antibiotics of 10∼40 mg/L showed an opportunity to inhibit respiration over a short time and the combined effect of these antibiotics exhibited a complex dose-dependence. The resistance was induced for the immobilized bacteria in the repeated measurement over a long-term period, which was fortunately solved by refreshing the biofilm via a nutrient supplementation. In the solution system, bacterial respirations were promoted by the tested antibiotics even at concentrations up to 160 mg/L, but β-glucosidase activity was inhibited by only 0.05 mg/L antibiotic. Cu²⁺ and Zn²⁺ were further inducted in the suspended system, and the combination of metals and antibiotics showed an obvious antagonistic effect. Real water detection indicated that the bacterial electrochemical method was expected to warn the water qualities with a sudden change by an appropriate testing condition.     

The Use of Seaweed and Sugarcane Bagasse for the Biological Treatment of Metal-contaminated Waters Under Sulfate-reducing Conditions

When wetlands reach maximum treatment capacity to remove heavy metals, removal can still take place through precipitation as sulfide because of the biological reduction of sulfate. To achieve this goal, anaerobic conditions must be attained, a sulfate source must exist, and an adequate substrate for sulfate-reducing bacteria (SRB) is also required. In the present work, two ligneous-cellulosic materials, a brown seaweed and sugarcane bagasse, have been selected as substrates for SRB growth. Experiments were simultaneously conducted in continuous operation in two columns (0.57 L each), one containing the ligneous-cellulosic material plus inoculum and another containing only the ligneous-cellulosic material. In this work, the removal of cadmium and zinc was studied because of their presence in effluents from mining/metallurgy operations. Results obtained indicated that the inoculated reactor was able to treat the effluent more efficiently than the noninoculated reactor considering the time course of the tests.     

Testing antimicrobials against biofilm bacteria

Standard laboratory methods are needed to assess the efficacy of antimicrobial agents that are applied to biofilm bacteria. Existing standard suspension tests and dried surface tests show much greater efficacy than antimicrobial agents applied to biofilms. The greater resistance of biofilm bacteria to antimicrobial agents can be attributed to a number of interacting factors, including reaction and diffusion processes that limit an agent's accessibility to bacteria, phenotypic changes in biofilm bacteria caused by stress, and adaptation of the bacteria. Because biofilm systems are so diverse, a variety of new biofilm tests with features that differ in important ways from existing tests will ultimately be required. For example, the biofilm test apparatus may include a pump and a continuous-flow stirred tank reactor. This report provides an overview of biofilm testing and suggests a strategy for creating standard test methods.     

Combining mixing regimes for optimized anaerobic wastewater treatment

Operational practice of high-rate anaerobic bioreactors such as upflow anaerobic sludge bed (UASB) reactors is generally based on maximization of the biomass concentration and, in the case of more than one reactor compartment, operation in parallel. In this article, a modeling approach is used to postulate that the treatment performance of anaerobic bioreactors can be improved by simple operational measures. To achieve minimized effluent soluble substrate concentrations, operation of two reactors in series combined with active exchange of biomass between both reactors is suggested. In this way, substrate concentrations lower than the minimum achievable concentration in a completely mixed reactor can be achieved. It is furthermore suggested that maximized biomass concentrations (and solid retention times [SRTs]) do not necessarily lead to minimized effluent concentrations of organic material. At elevated SRTs, the soluble microbial products resulting from biomass turnover are shown to represent the main fraction of soluble organic material in the effluent of the reactor, limiting treatment efficiency.     

The Use of Thin-Layer Chromatography in Experimental Xenobiology

Abstract

Studies of the metabolism and disposition of most drugs or other xenobiotics share one common feature: the need for an analytical method to measure the xenobiotic and/or its biotransformation products in biosamples. Although chromatographic methods such as GC and HPLC are important and preferred by some laboratories, initial investigations of new xenobiotic entities are often conducted using radiolabeled compounds, and, in such instances, TLC assay methods are frequently selected.

TLC is utilized in xenobiotic metabolism and disposition studies to evaluate the radiochemical purity of labeled xenobiotics, the extent of metabolism, the profile of metabolites in blood and excreta, the concentrations of the xenobiotic and specific metabolites in blood and other biosamples, the stability of the xenobiotic and its metabolites in biosamples, and the relative behavior of metabolites and reference compounds. Preparative TLC is frequently used to isolate specific metabolites. Thin-layer radiochromatography (TLRC) has been utilized to study the pharma-cokinetics of drugs and other xenobiotics in man and in animals. Especially where radiolabeled compounds are used, TLC provides an attractive chromatographic alternative to GC and HPLC in research in experimental xenobiology.     

Adsorption behavior of mercury and precious metals on cross-linked chitosan and the removal of ultratrace amounts of mercury in concentrated hydrochloric acid by a column treatment with cross-linked chitosan.

Cross-linked chitosan was synthesized with chitosan and ethylene glycol diglycidyl ether. The adsorption behavior of trace amounts of metal ions on the cross-linked chitosan was systematically examined by packing it in a mini-column, passing a metal solution through it and measuring metal ions in the effluent by ICP-MS. The cross-linked chitosan adsorbed mercury and precious metals (Pd, Pt, and Au) at pH values from acidic to neutral. Especially, mercury in concentrated hydrochloric acids could be adsorbed on cross-linked chitosan quantitatively by an anion-exchange mechanism in the form of a stable chloride complex. This method was applied to the removal of mercury from commercially available hydrochloric acid; more than 97% of mercury was removed, and the residual mercury in the hydrochloric acid (Grade: for trace analysis) was found to be 0.15 ppb. Mercury adsorbed on the cross-linked chitosan could be easily desorbed with an eluent containing I M hydrochloric acid and 0.05 M thiourea. The thus-refreshed cross-linked chitosan could be repeatedly used for the removal of mercury in hydrochloric acid.     

The effect of imposed flux on biofouling in reverse osmosis: Role of concentration polarisation and biofilm enhanced osmotic pressure phenomena

This paper describes a systematic study of biofouling in reverse osmosis process using model bacteria of Pseudomonas fluorescens and employing a sodium chloride tracer response technique for fouling characterization. It was found that the growth of biofilm at constant flux following initial bacteria colonization of the membrane surface increased with imposed flux. The rationale was that biofilm growth was nutrient dependent, where the nutrient availability at the membrane wall was controlled by the magnitude of concentration polarization, which is driven by flux. The salt tracer response showed that the biofouling comprised a hydraulic resistance and induced an enhanced osmotic pressure phenomenon; known as the biofilm enhanced osmotic pressure (BEOP) effect [M. Herzberg, M. Elimelech, Biofouling of reverse osmosis membranes: role of biofilm-enhanced osmotic pressure, Journal of Membrane Science 295 (2007) 11–20], due to hindered back diffusion of solutes through the tortuous path of the heterogeneous structure of the biofilm. For the conditions studied, the contribution of BEOP to transmembrane pressure increase was greater than the hydraulic resistance.     

Influence of Operational Parameters and Low Nickel Concentrations on Partial Nitrification in a Submerged Biofilter

The effect of Ni(2+) concentrations on ammonium oxidation was studied in a batch and partial bionitrification reactor (PBNR). The nitrification rates up to the concentration of 0.1 mg Ni(2+)/l were close to those without Ni(2+). After testing the operational conditions in the PNBR, the highest NO(2)-N/NO( x )-N ratio was achieved at the DO concentrations of 2.0 mg/l and pH 9.00. The PNBR was operated at steady state (NH(4)-N loading rate and NO(2)-N/NO( x )-N ratio were 405?g m(-3) day(-1) and 0.74, respectively) before exposure to Ni(2+). The removal efficiency of NH(4)-N and NO(2)-N/NO( x )-N ratio in the effluent waters was increased by adding low concentrations of heavy metals to the PBNR. The average number of aerobic mesophilic bacteria at the biofilm surface and in the water in the void volume of PNBR were 1.0?×?10(4) CFU/g and 1.4?×?10(5) CFU/ml, respectively.     

Energy generation by methanation of persistent wastes

A 15-L anaerobic fixed-film reactor (AFFR) was evaluated for treating a trade effluent containing inhibitory concentrations of persistent branched-chain fatty acids, namely 2-ethylhexanoic acid (2-EHA) and neopentanoic acid (NPA), at a total of 17,000 mg COD/L. The AFFR was packed with fire-expanded clay spheres, and start-up was accomplished in 60 d. The organic load was increased in steps from 1.1 to 8.5 g COD/L/d. Total COD, 2-EHA, and NPA removal efficiencies were maintained above 70, 98, and 75%, respectively. The reactor could recover from a shock load of 150% increase in organic load. Combined mechanisms of organic adsorption and biodegradation rendered the AFFR more stable with shock loads. Mathane gas produced from the process could be used for preheating the effluent.