Effects of different sources of copper on Ctr1, ATP7A,ATP7B,MT and DMT1 protein and gene expression in Caco-2 cells

Copper sulfate (CuSO₄), micron copper oxide (micron CuO) and nano copper oxide (nano CuO) at different concentrations were, respectively, added to culture media containing Caco-2 cells and their effects on Ctr1, ATP7A/7B, MT and DMT1 gene expression and protein expression were investigated and compared. The results showed that nano CuO promoted mRNA expression of Ctr1 in Caco-2 cells, and the difference was significant compared with micron CuO and CuSO₄. Nano CuO was more effective in promoting the expression of Ctr1 protein than CuSO₄ and micron CuO at the same concentration. Nano CuO at a concentration of 62.5 μM increased the mRNA expression levels of ATP7A and ATP7B, and the difference was significant compared with CuSO₄. The addition of CuSO₄ and nano CuO to the culture media promoted the expression of ATP7B proteins. CuSO₄ at a concentration of 125 μM increased the mRNA expression level of MT in Caco-2 cells, and the difference was significant compared with nano CuO and micron CuO. Nano CuO at a concentration of 62.5 μM inhibited the mRNA expression of DMT1, and the difference was significant compared with CuSO₄ and micron CuO. Thus, the effects of CuSO₄, micron CuO and nano CuO on the expression of copper transport proteins and the genes encoding these proteins differed considerably. Nano CuO has a different uptake and transport mechanism in Caco-2 cells to those of CuSO₄ and micron CuO.     

Biomass accumulation and carbon utilization in layered sand filter biofilm systems receiving milk fat and detergent mixtures

This research evaluated utilization of organic compounds and biomass accumulation in three different layered sand filter biofilm systems renovating a mixture of detergent and milk fat. Organic compounds were measured as chemical oxygen demand (COD) and biomass as ATP. A two-layer sand filter with coarse sand on the top and fine sand at the bottom; a three-layer sand filter with pea gravel on the top, coarse sand in the middle, and fine sand at the bottom; and a two-filters in-series sand filter biofilm system each received mixtures of butterfat and detergent. While the three-layer sand filter system had greater COD utilization capacity due to a longer filter run, the two-filters in-series sand filter system had greater COD utilization efficiency. The COD utilization was highly associated with the period of filter run to clogging. The decrease of COD utilization rate could lead to the clogging of sand filters. To maintain long filter runs without clogging, a COD loading rate of 2100 mg O(2)/m(2)/h was recommended for three-layer sand filters and 1800 mg O(2)/m(2)/h for two-layer sand filters for high fat content wastewater. In the two-layer sand filters and the three-layer sand filters, biomass did not seem to accumulate in any particular layer on a unit volume of sand. However, based on unit surface area, the coarse sand layer accumulated more biomass than the fine sand layer and the pea gravel layer.     

An investigation into the microbial clogging potential of selected filter media as a result of biodegradation of a high-strength sulphate-rich alkaline leachate

The research examines the potential for bio-clogging in filter packs containing fine sand of the type typically used in extraction wells for pumping leachates containing fine particulate matter, such as cement kiln dust (CKD). Three filter media with different particle sizes were used: 1.7–4.75, 0.35–1.0, and 0.235–0.45 mm. Each sand filter was tested using a leachate recirculating column reactor with a free drainage layer, on top of which was placed the filtration medium which was kept saturated and at a positive hydrostatic head by a 2-l reservoir of leachate. The leachate was collected from a landfill site that had been used for the co-disposal of municipal solid waste (MSW) and CKD. The leachate used was filtered by passing through a Whatman GFA filter paper before being added to the reactors in order to eliminate as far as possible the non-biological clogging which might have resulted from the introduction of particulate matter in the form of CKD. The filters and a control experiment were run under anaerobic conditions at 35 °C. The bio-clogging potential was observed by taking differential manometer readings from manometers located in the drainage and reservoir sections of the reactor. No clogging was detected using the coarser of the filter media, but there was some clogging when a finer filter medium was used. Head space gas analysis indicated that methanogenic activity was inhibited and analysis of the liquid phase indicated that the microbial process responsible for removal of chemical oxygen demand (COD) was principally one of sulphate reduction.     

Bacterial clogging of porous media: A new modelling approach

A comparison is made between existing mathematical models and experimental data that relate the reduction of the saturated hydraulic conductivity (K) of a porous medium to the porosity reduction caused by microbial growth. The models yielded a realistic prediction of a data set obtained with a model porous medium consisting of millimeter‐size glass spheres, but failed to predict the clogging behaviour observed in smaller‐than‐1‐mm sand. A new modelling approach, semi‐mechanistic in nature, is proposed that gives good predictions of fine sand media as well. It relaxes the assumption about uniformly‐thick biofilms by allowing a second arrangement to occur, i.e. discrete plugs filling the pore lumen. The new model requires input data on two intrinsic properties of the system, which renders it sufficiently flexible as to fit very different data sets. The two model parameters are K_min, the minimum K value when all porosity is filled with microorganisms, and B_c, the biovolume fraction at which most cell detachment from biofilm occurs.     

A Limited Microbial Consortium Is Responsible for Extended Bioreduction of Uranium in a Contaminated Aquifer

Subsurface amendments of slow-release substrates (e.g., emulsified vegetable oil [EVO]) are thought to be a pragmatic alternative to using short-lived, labile substrates for sustained uranium bioimmobilization within contaminated groundwater systems. Spatial and temporal dynamics of subsurface microbial communities during EVO amendment are unknown and likely differ significantly from those of populations stimulated by soluble substrates, such as ethanol and acetate. In this study, a one-time EVO injection resulted in decreased groundwater U concentrations that remained below initial levels for approximately 4 months. Pyrosequencing and quantitative PCR of 16S rRNA from monitoring well samples revealed a rapid decline in groundwater bacterial community richness and diversity after EVO injection, concurrent with increased 16S rRNA copy levels, indicating the selection of a narrow group of taxa rather than a broad community stimulation. Members of the Firmicutes family Veillonellaceae dominated after injection and most likely catalyzed the initial oil decomposition. Sulfate-reducing bacteria from the genus Desulforegula, known for long-chain fatty acid oxidation to acetate, also dominated after EVO amendment. Acetate and H₂ production during EVO degradation appeared to stimulate NO₃⁻, Fe(III), U(VI), and SO₄²⁻ reduction by members of the Comamonadaceae, Geobacteriaceae, and Desulfobacterales. Methanogenic archaea flourished late to comprise over 25% of the total microbial community. Bacterial diversity rebounded after 9 months, although community compositions remained distinct from the preamendment conditions. These results demonstrated that a one-time EVO amendment served as an effective electron donor source for in situ U(VI) bioreduction and that subsurface EVO degradation and metal reduction were likely mediated by successive identifiable guilds of organisms.     

Effects of Light Crude Oil Contamination on the Physical and Mechanical Properties of Fine Sand

The use of oil-contaminated sand in building and construction is now being considered as an alternative and cost-effective way to minimize its adverse effect on the environment. To achieve this, the effect of oil contamination on the important mechanical properties of sand should be investigated first. This study investigated the effect of petroleum-derived contaminants on the water absorption, permeability, cohesion, friction angle, and shear strength of fine sand. Contaminated samples were prepared by mixing fine sand with different percentages of light crude oil (0 to 20%). The results indicated that the water absorption of fine sand decreases with an increase in crude oil. An increase in the cohesion was observed for sand with up to 1% of oil contamination, after which the cohesion began to decrease, which also results in the reduction in the permeability. A slight reduction in the friction angle was found for oil-contaminated fine sand. At a low normal stress of 50 kPa, as the percentage of light crude oil increased, the shear strength increased up to 1% of oil contamination and then it decreased. These results provided useful information on how oil-contaminated sand can be used safely and effectively in building and construction.     

Lipase production by Aspergillus ibericus using olive mill wastewater

Olive mill wastewater (OMW) characteristics make it a suitable resource to be used as a microbial culture media to produce value-added compounds, such as enzymes. In this work, the ability of the novel species Aspergillus ibericus to discolor OMW and produce lipase was studied. An initial screening on plates containing an OMW-based agar medium and an emulsified olive oil/rhodamine-B agar medium was employed to select the strain A. ibericus MUM 03.49. Then, experiments in conical flasks with liquid OMW-based media showed that the fungus could growth on undiluted OMW, with a chemical oxygen demand (COD) of 97 ± 2 g/L, and to produce up to 2,927 ± 54 U/L of lipase. When pure OMW was used in the media, the maximum COD and color reduction achieved were 45 and 97 %, respectively. When OMW diluted to 10 % was used, A. ibericus was able to reduce phenolic and aromatic compounds by 37 and 39 %, respectively. Additionally, lipase production was found to be promoted by the addition of mineral nutrients. When the fermentations were scaled up to a 2-L bioreactor, A. ibericus produced up to 8,319 ± 33 U/L of lipase, and the maximum COD and color reduction were 57 and 24 %, respectively.     

Removal of oil by walnut shell media

Studies were conducted to evaluate the oil sorption capacities of walnut shell media. Sorption capacity is the weight of oil picked up by unit weight of a sorbent. Initial oil pick-up by walnut shell media on pure oil and oil on aqueous medium was evaluated. Batch kinetic studies were conducted to evaluate the equilibrium time required by walnut shell media for sorbing oil. For pure oil medium, sorption capacities of 0.30g/g, 0.51g/g and 0.58g/g were obtained for standard mineral oil, vegetable oil and DoALL Bright-Edge oil, respectively. The results showed sorption capacities of 0.56g/g, 0.58g/g and 0.74g/g for standard mineral oil, vegetable oil and DoALL Bright-Edge oil, respectively, for oil on aqueous medium. It was found that sorbed oil could be recovered from walnut shell media by applying pressure. The study showed that walnut shell media can be used as a sorbent for oil removal.     

Network succession reveals the importance of competition in response to emulsified vegetable oil amendment for uranium bioremediation

Discerning network interactions among different species/populations in microbial communities has evoked substantial interests in recent years, but little information is available about temporal dynamics of microbial network interactions in response to environmental perturbations. Here, we modified the random matrix theory‐based network approach to discern network succession in groundwater microbial communities in response to emulsified vegetable oil (EVO) amendment for uranium bioremediation. Groundwater microbial communities from one control and seven monitor wells were analysed with a functional gene array (GeoChip 3.0), and functional molecular ecological networks (fMENs) at different time points were reconstructed. Our results showed that the network interactions were dramatically altered by EVO amendment. Dynamic and resilient succession was evident: fairly simple at the initial stage (Day 0), increasingly complex at the middle period (Days 4, 17, 31), most complex at Day 80, and then decreasingly complex at a later stage (140–269 days). Unlike previous studies in other habitats, negative interactions predominated in a time‐series fMEN, suggesting strong competition among different microbial species in the groundwater systems after EVO injection. Particularly, several keystone sulfate‐reducing bacteria showed strong negative interactions with their network neighbours. These results provide mechanistic understanding of the decreased phylogenetic diversity during environmental perturbations.     

Dynamic Succession of Groundwater Functional Microbial Communities in Response to Emulsified Vegetable Oil Amendment during Sustained In Situ U(VI) Reduction

A pilot-scale field experiment demonstrated that a one-time amendment of emulsified vegetable oil (EVO) reduced groundwater U(VI) concentrations for 1 year in a fast-flowing aquifer. However, little is known about how EVO amendment stimulates the functional gene composition, structure, and dynamics of groundwater microbial communities toward prolonged U(VI) reduction. In this study, we hypothesized that EVO amendment would shift the functional gene composition and structure of groundwater microbial communities and stimulate key functional genes/groups involved in EVO biodegradation and reduction of electron acceptors in the aquifer. To test these hypotheses, groundwater microbial communities after EVO amendment were analyzed using a comprehensive functional gene microarray. Our results showed that EVO amendment stimulated sequential shifts in the functional composition and structure of groundwater microbial communities. Particularly, the relative abundance of key functional genes/groups involved in EVO biodegradation and the reduction of NO₃⁻, Mn(IV), Fe(III), U(VI), and SO₄²⁻ significantly increased, especially during the active U(VI) reduction period. The relative abundance for some of these key functional genes/groups remained elevated over 9 months. Montel tests suggested that the dynamics in the abundance, composition, and structure of these key functional genes/groups were significantly correlated with groundwater concentrations of acetate, NO₃⁻, Mn(II), Fe(II), U(VI), and SO₄²⁻. Our results suggest that EVO amendment stimulated dynamic succession of key functional microbial communities. This study improves our understanding of the composition, structure, and function changes needed for groundwater microbial communities to sustain a long-term U(VI) reduction.     

Xanthan degragation by biofilm in porous media

Biological activity in oil reservoirs can cause significant problems such as souring and plugging. This study focuses on the problem of polymer degradation and permeability reduction due to biofilm formation during polymer injection for improved oil recovery. Polymers are included in injection fluids to increase their viscosity. Results relating biological processes and polymer degradation to fluid‐dynamic conditions in a laboratory model porous medium are presented.

A transparent flow cell with an etched two‐dimensional network of pores served as a model porous medium. A sterile xanthan polymer and natural sea water solution were continuously injected into the porous medium. A bacterial culture capable of xanthan degradation was introduced into the cell by a single injection. Some of the cells from this culture attached to the pore walls forming an immobile bacterial culture, termed biofilm. The development of this biofilm, its xanthan degradation and its effect on permeability were measured.

The effects of injection rate and rate transitions were analyzed. Injection fluid viscosity was reduced by 30% after 5 min flow through the porous medium at the maximum steady state degradation rate observed. Permeability was significantly reduced by the xanthan degrading biofilm, causing an increase in pressure drop through the porous medium of up to 80%. Polymer injection in oil reservoirs may, therefore, have negative effects on oil recovery, unless efficient biofouling control is applied. The methodology presented may serve as a tool in the development of biofouling control measures in porous media.     

Development of an improved anaerobic filter for municipal wastewater treatment

Development of an improved reactor configuration of anaerobic filter was carried out for the elimination of clogging of filter media. The experiments over different hydraulic retention times (HRTs) indicated that the HRT of 12 h was the most appropriate one for the system studied while treating the municipal wastewater, which resulted 90% and 95% BOD and COD reduction, respectively. Reduction up to 95% in suspended solids concentration could be achieved without any pretreatment. The specific biogas yield obtained was 0.35 m(3) CH(4)/kgCODr with 70% of CH(4) content in the biogas generated from the system at the HRT of 12 h. Operational problems such as clogging of filter media were not observed throughout the period of study over 600 d.     

Studies of the Cell Surface Properties of <Emphasis Type="Italic">Candida</Emphasis> Species and Relation to the Production of Biosurfactants for Environmental Applications

In practical bioremediation of petroleum pollution, treatment systems often use soil, sand, and other aquifer porous media besides water solutions. The distribution of the microbial cell also plays an important role in the whole process of bioremediation; therefore, the adhesion ability of cells to porous media is one of the key factors influencing the efficiency of treatment. The probable modes of hydrocarbon uptake in cells of Candida were studied based on data for cell hydrophobicity, emulsifying activity, surface tension, and interfacial tension of the cell-free culture medium. Six Candida strains were cultivated in insoluble and soluble substrates for 144 h, including n-hexadecane, soybean oil, ground-nut oil refinery residue, corn steep liquor, and glucose. The results obtained showed the potential of yeasts for application in the removal of hydrophobic compounds. Depending the strain and substrate used the adhesion ability of yeast cells and the production of surfactants and emulsifiers can take place simultaneously, thus increasing the efficiency of bioremediation treatment of petroleum pollution. The application of crude biosurfactants separated from the yeast cells was also demonstrated by tests of removal of petroleum and the derivate motor oil adsorbed in sand samples. Biosurfactants produced in low-cost medium were able to remove 90% of the hydrophobic contaminants.     

模拟油藏条件下内源微生物群落空间分布规律

【背景】油藏内源微生物群落是开展内源微生物驱油技术的物质基础,由于油藏多孔介质取样技术难度大、成本高,实施内源微生物驱油后从注入端到产出端多孔介质中的内源微生物空间分布规律尚不明确。【目的】通过室内长岩心连续驱替实验模拟油藏内源微生物驱油过程,分析实施后不同空间位点油砂上吸附的内源微生物群落结构,揭示从注入端到产出端内源微生物群落的空间分布规律。【方法】借助高通量测序技术及荧光定量PCR技术解析不同空间位点油砂原位微生物群落信息。【结果】注入端到产出端不同空间位点生态环境的差异及菌属间的相互作用造成油藏内源微生物群落空间分布差异,存在明显的好氧、厌氧空间演替变化规律。岩心前端主要存在一些好氧类的产生物表面活性剂类微生物如假单胞菌属,岩心中部主要存在兼性和厌氧类的微生物如地芽孢杆菌、厌氧杆菌属,岩心末端主要分布严格厌氧类细菌和产甲烷古菌,厌氧类微生物代谢产生的H2、CO2和乙酸分子可以为产甲烷古菌提供代谢底物。【结论】通过室内物模油砂研究,首次明确了内源微生物群落在多孔介质中从注入端到产出端的空间分布规律,证实油藏内源微生物的好氧、厌氧空间接替分布规律,深化了对油藏内源微生物的认识。     

Towards optimum permeability reduction in porous media using biofilm growth simulations

While biological clogging of porous systems can be problematic in numerous processes (e.g., microbial enhanced oil recovery—MEOR), it is targeted during bio‐barrier formation to control sub‐surface pollution plumes in ground water. In this simulation study, constant pressure drop (CPD) and constant volumetric flow rate (CVF) operational modes for nutrient provision for biofilm growth in a porous system are considered with respect to optimum (minimum energy requirement for nutrient provision) permeability reduction for bio‐barrier applications. Biofilm growth is simulated using a Lattice‐Boltzmann (LB) simulation platform complemented with an individual‐based biofilm model (IbM). A biomass detachment technique has been included using a fast marching level set (FMLS) method that models the propagation of the biofilm–liquid interface with a speed proportional to the adjacent velocity shear field. The porous medium permeability reduction is simulated for both operational modes using a range of biofilm strengths. For stronger biofilms, less biomass deposition and energy input are required to reduce the system permeability during CPD operation, whereas CVF is more efficient at reducing the permeability of systems containing weaker biofilms. Biotechnol. Bioeng. 2009;103: 767–779. © 2009 Wiley Periodicals, Inc.     

Biofilm development and the dynamics of preferential flow paths in porous media

A two-dimensional pore-scale numerical model was developed to evaluate the dynamics of preferential flow paths in porous media caused by bioclogging. The liquid flow and solute transport through the pore network were coupled with a biofilm model including biomass attachment, growth, decay, lysis, and detachment. Blocking of all but one flow path was obtained under constant liquid inlet flow rate and biomass detachment caused by shear forces only. The stable flow path formed when biofilm detachment balances growth, even with biomass weakened by decay. However, shear forces combined with biomass lysis upon starvation could produce an intermittently shifting location of flow channels. Dynamic flow pathways may also occur when combined liquid shear and pressure forces act on the biofilm. In spite of repeated clogging and unclogging of interconnected pore spaces, the average permeability reached a quasi-constant value. Oscillations in the medium permeability were more pronounced for weaker biofilms.     

Chemical and Physical Factors Influencing Growth of Phytophthora parasitica var. nicotianae on Vegetable Oils

Factors influencing the growth of Phytophthora parasition var. nicotiona on vegetable oil liquid media were studied. This fungus grew poorly on oleic acid, triolein, and tristearin. Cholesterol and α-tocopherol reversed the toxicity of oleic acid. Growth was often stimulated by addition of mineral oil to a glucose medium, and sterols dissolved in mineral oil frequently stimulated grwoth. Shaking produced a 5-fold or more increase in growth on either glucose or cottonseed oil media. Cholesterol (20 μ/ml further increased growth on glucose meidum in shake culture by at least 100%. Growth was usually better at pH 6.7 than at 5.4 or 7.7. Growth in shake culture on a glucose medium supplemented with cholesterol and α-tocopherol approached that occuring on vegetable oil media.     

Reduction of porous media permeability from in situ Leuconostoc mesenteroides growth and dextran production

In situ growth of bacteria in a porous medium can alter the permeability of that media. This article reveals that the rate of permeability alteration can be controlled by the inoculation strategy, nutrient concentrations, and injection rates. Based on experimental observations a phenomenological model has been developed to describe the inoculation of the porous medium, the in situ growth of bacteria, and the permeability decline of the porous medium. This model consists of two phases that describe the bacteria in the porous medium: (1) the nongrowth phase in which cell transport and retention are occurring; and (2) the growth phase in which the retained cells grow and plug the porous media. Transition from the transport phase to the growth phase is governed by the growth lag time of the cells within the porous medium. The importance of the inoculum injection strategy and the nutrient injection strategy is illustrated by the model. (c) 1996 John Wiley & Sons, Inc.     

Effect of biological activity on the movement of fluids through porous rocks and sediments and its application to enhanced oil recovery

Movement of fluids through geological porous media is an important factor in ore genesis, crude oil genesis, and in oil recovery from oil reservoirs. Such a movement may be beneficially affected by biological in situ activity. We have shown that once bacteria have been introduced into a porous medium, they are able to penetrate into their environment and multiply without decreasing significantly the rock's permeability. The limiting bacterial concentration, which may cause plugging, depends on the bacterial size, pore size, and pore size distribution of the reservoir rock. Microorganisms may, under anaerobic conditions, affect the composition of crude oil by increasing its asphaltenic fraction. In simulated systems microorganisms increased the recovery of the oil in place approximately five times. A bacterial consortium was more effective than a single pure bacterial strain.     

Application of the growth substrate pulsed feeding technique to a process of chloroform aerobic cometabolism in a continuous-flow sand-filled reactor

A process of chloroform (CF) aerobic cometabolic biodegradation by a butane-growing consortium was studied for 354 days in a 2-m continuous-flow sand-filled reactor. The study was aimed at (a) investigating the oxygen/substrate pulsed injection as a tool to control the clogging of the porous medium, to attain a wide bioreactive zone and to reduce substrate inhibition on CF cometabolism; (b) developing a reliable model of CF cometabolism in porous media. While the continuous supply of butane rapidly led to the clogging of the porous medium due to excessive biomass growth, the testing of six types of oxygen/substrate pulsed feeding led to the identification of a feeding schedule capable to prevent aquifer clogging and to lead to the development of a long bioreactive zone and to satisfactory CF degradation performances. The tested model of aerobic cometabolism allowed a suitable interpretation of the experimental data as long as the ratio of CF degraded to butane consumed was ≤0.27 mg_CF mg_butane^?1. A long-term 1-D simulation of the best-performing schedule of pulsed oxygen/substrate supply extended to a 30-m aquifer length resulted in a 20-m bioreactive zone and in a 96% CF removal.