Search

Bacterial deposition and transport are key to microbial ecology and biotechnological applications. We therefore tested whether electrokinetic forces (electroosmotic shear force (FEOF), electrophore...

Phages (i.e., viruses infecting bacteria) are considered to be good indicators and tracers for fecal pollution, hydraulic flow, or colloidal transport in the subsurface. They are typically quantified as total virus particles (VLP) or plaque forming units (PFU) of infectious phages. As transport may lead to phage deactivation, VLP quantification can overestimate the number of infectious phages. In contrast, PFU counts may underestimate the transport of total virus particles. Using PFU and tunable resistive pulse sensing-based counting for active and total phages, respectively, we quantified the effect of transport through laboratory percolation columns on the specific infectivity (SI). The SI is defined by the ratio of total VLP to PFU and is a measure for the minimum particle numbers needed to create a single infection. Transport of three marine tracer phages and the coli-phage (T4) was described by colloidal filtration theory. We found that apparent collision efficiencies of active and total phages differed. Depending on the phage properties (e.g., morphology or hydrophobicity), passage through a porous medium led to either an increasing or decreasing SI of effluent phages. Our data suggest that both phage mass recovery and the SI should be considered in quantitative phage tracer experiments.

Although several studies examined the transport of viruses in terrestrial systems only few studies exist on the use of marine phages (i.e., nonterrestrial viruses infecting marine host bacteria) as sensitively detectable microbial tracers for subsurface colloid transport and water flow. Here, we systematically quantified and compared for the first time the effects of size, morphology and physicochemical surface properties of six marine phages and two coliphages (MS2, T4) on transport in sand-filled percolated columns. Phage-sand interactions were described by colloidal filtration theory and the extended Derjaguin-Landau-Verwey-Overbeek approach (XDLVO), respectively. The phages belonged to different families and comprised four phages never used in transport studies (i.e., PSA-HM1, PSA-HP1, PSA-HS2, and H3/49). Phage transport was influenced by size, morphology and hydrophobicity in an approximate order of sizehydrophobicity ≥ morphology. Two phages PSA-HP1, PSA-HS2 (Podoviridae and Siphoviridae) exhibited similar mass recovery as commonly used coliphage MS2 and were 7-fold better transported than known marine phage vB_PSPS-H40/1. Differing properties of the marine phages may be used to trace transport of indigenous viruses, natural colloids or anthropogenic nanomaterials and, hence, contribute to better risk analysis. Our results underpin the potential role of marine phages as microbial tracer for transport of colloidal particles and water flow.

Microbial biofilms can cause severe problems in technical installations where they may give rise to microbially influenced corrosion and clogging of filters and membranes or even threaten human health, e.g. when they infest water treatment processes. There is, hence, high interest in methods to prevent microbial adhesion as the initial step of biofilm formation. In environmental technology it might be desired to enhance bacterial transport through porous matrices. This motivated us to test the hypothesis that the attractive interaction energy allowing cells to adhere can be counteracted and overcome by the shear force induced by electroosmotic flow (EOF, i.e. the water flow over surfaces exposed to a weak direct current (DC) electric field). Applying EOF of varying strengths we quantified the deposition of Pseudomonas fluorescens Lp6a in columns containing glass collectors and on a quartz crystal microbalance. We found that the presence of DC reduced the efficiency of initial adhesion and bacterial surface coverage by85%. A model is presented which quantitatively explains the reduction of bacterial adhesion based on the extended Derjaguin, Landau, Verwey, and Overbeek (XDLVO) theory of colloid stability and the EOF-induced shear forces acting on a bacterium. We propose that DC fields may be used to electrokinetically regulate the interaction of bacteria with surfaces in order to delay initial adhesion and biofilm formation in technical installations or to enhance bacterial transport in environmental matrices.

Mycelia have been recently shown to actively transport polycyclic aromatic hydrocarbons (PAH) in water-unsaturated soil over the range of centimeters, thereby efficiently mobilizing hydrophobic PAH beyond their purely diffusive transport in air and water. However, the question if mycelia-based PAH transport has an effect on PAH biodegradation was so far unsolved. To address this, we developed a laboratory model microcosm mimicking air-water interfaces in soil. Chemical analyses demonstrated transport of the PAH fluorene (FLU) by the mycelial oomycete Pythium ultimum that was grown along the air-water interfaces. Furthermore, degradation of mycelia-transported FLU by the bacterium Burkholderia sartisoli RP037-mChe was indicated. Since this organism expresses eGFP in response to a FLU flux to the cell, it was also as a bacterial reporter of FLU bioavailability in the vicinity of mycelia. Confocal laser scanning microscopy (CLSM) and image analyses revealed a significant increase of eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU. Hence, we could show that physically separated FLU becomes bioavailable to bacteria after transport by mycelia. Experiments with silicon coated glass fibers capturing mycelia-transported FLU guided us to propose a three-step mechanism of passive uptake, active transport and diffusion-driven release. These experiments were also used to evaluate the contributions of these individual steps to the overall mycelial FLU transport rate.

To cope with heterogeneous subsurface environments mycelial microorganisms have developed a unique ramified growth form. By extending hyphae, they can obtain nutrients from remote places and transport them even through air gaps and in small pore spaces, repectively. To date, studies have been focusing on the role that networks play in the distribution of nutrients. Here, we investigated the role of mycelia for the translocation of nonessential substances, using polycyclic aromatic hydrocarbons (PAHs) as model compounds. We show that the hyphae of the mycelial soil oomycete Pythium ultimum function as active translocation vectors for a wide range of PAHs. Visualization by two-photon excitation microscopy (TPEM) demonstrated the uptake and accumulation of phenanthrene (PHE) in lipid vesicles and its active transport by cytoplasmic streaming of the hyphae ('hyphal pipelines'). In mycelial networks, contaminants were translocated over larger distances than by diffusion. Given their transport capacity and ubiquity, hyphae may substantially distribute remote hydrophobic contaminants in soil, thereby improving their bioavailability to bacterial degradation. Hyphal contaminant dispersal may provide an untapped potential for future bioremediation approaches.

A test kit based on living, lyophilized bacterial bioreporters emitting bioluminescence as a response to arsenite and arsenate was applied during a field campaign in six villages across Bangladesh. Bioreporter field measurements of arsenic in groundwater from tube wells were in satisfying agreement with the results of spectroscopic analyses of the same samples conducted in the lab. The practicability of the bioreporter test in terms of logistics and material requirements, suitability for high sample throughput, and waste disposal was much better than that of two commercial chemical test kits that were included as references. The campaigns furthermore demonstrated large local heterogeneity of arsenic in groundwater, underscoring the use of well switching as an effective remedy to avoid high arsenic exposure.

The hypothesis that dissolved organic carbon (DOC) enhances the mass transfer of hydrophobic organic compounds from nonaqueous phase liquids (NAPLs) into the aqueous phase above that attributable to dissolved molecular diffusion alone was tested. In controlled experiments, mass transfer rates of five NAPL-phase PAHs (log K(OW) 4.15-5.39) into the aqueous phase containing different concentrations of DOC were measured. Mass transfer rates were increased by up to a factor of 4 in the presence of DOC, with the greatest enhancement being observed for more hydrophobic compounds and highest DOC concentrations. These increases could not be explained by dissolved molecular diffusion alone, and point to a parallel DOC-mediated diffusive pathway. The nature of the DOC-mediated diffusion pathway as a function of the DOC concentration and PAH sorption behavior to the DOC was investigated using diffusion-based models. The DOC-enhanced mass transfer of NAPL-phase hydrophobic compounds into the aqueous phase has important implications for their bioremediation as well as bioconcentration and toxicity.

Speciation affects trace metal bioavailability. One model used to describe the importance of speciation is the biotic ligand model (BLM), wherein the competition of inorganic and organic ligands with a biotic ligand for free-ion trace metal determines the ultimate metal availability to biota. This and similar models require natural ligand concentrations and conditional stability constants as input parameters. In concept, the BLM is itself an analogue of some analytical approaches to the determination of trace metal speciation. A notable example is competitive ligand equilibration/cathodic stripping voltammetry, which employs an artificial ligand for comparative assessment of natural ligand concentrations and discrete conditional stability constants (i.e., BLM parameters) in a natural sample. Here, we report a new numerical approach to voltammetric speciation and parameter estimation that employs multiple analytical windows and a two-step optimization process, simultaneously generating both parameters and a complete suite of corresponding species concentrations. This approach is more powerful, systematic, and flexible than those previously reported.

Bioremediation relies on the stability of enzymatic activities, particularly when bioavailable contaminant concentrations do not permit much renewal of microbial biomass. Starving Delftia acidovorans MC1 were found to lose specific degradation activity, while accumulating variants of the alpha-ketoglutarate-dependent dioxygenase RdpA, the enzyme initiating the degradation of (RS)-2-(2,4-dichlorophenoxy)propionate. These variants differed in their pI and originated from post-translational modification, since there is only one rdpA gene in the genome. It was tested if RdpA modification resulted from carbonylation by reactive oxygen species, known side products of dioxygenase reactions. Carbonylated amino acids in proteins of starved cells were specifically derivatized with 2,4-dinitrophenylhydrazine. Subsequent immunolabeling of the resulting hydrazones and mass spectrometry of tryptic digests confirmed different levels of carbonylation of RdpA.

The hypothesis that humic acids (HA) can act as carriers of polycyclic aromatic hydrocarbon (PAH) compounds and provide bacteria with PAH fluxes beyond those obtained by diffusion of the water-dissolved fraction of these poorly soluble chemicals was tested. Controlled degradation experiments using microcrystalline phenanthrene and Sphingomonas sp. LH162 were performed without HA and in the presence of two HA concentrations. In the absence of HA, phenanthrene was steadily present at the maximum dissolved concentration, indicating that the phenanthrene dissolution flux compensated for the consumption. This was confirmed by dissolution experiments where phenanthrene dissolved roughly an order of magnitude faster than it was consumed in the degradation experiments. Interestingly, the presence of HA further increased the rates of phenanthrene degradation by factors up to 4.8 in an HA concentration-dependent manner. This can only be interpreted by an HA-mediated transport of phenanthrene to the cells, supplementing diffusive uptake from the freely dissolved phase. It is proposed that HA-sorbed phenanthrene is released directly to the cells upon their interaction with HA aggregates, increasing the total phenanthrene flux and also degradation.

There is growing interest in employing electro-bioremediation, a hybrid technology of bioremediation and electrokinetics for the treatment of contaminated soil. Most present applications of electrokinetics aim at pollutant extraction, which requires transport over large distances facilitated by electroosmotic flow (EOF). They do not explicitly account for the possibility that EOF passing along soil particles stimulates the release of hydrophobic organic compounds (HOC) and locally improves pollutant bioavailability. Here, we report on the stimulated release of polycyclic aromatic hydrocarbon (phenanthrene) from model organic matter in the presence of direct current (DC)-electric fields (0.5-2 V cm(-1)) typically used in electrobioremediation measures. Alginate beads were employed as a model polymer release system (MPRS) exhibiting similar release behavior as natural organic matter (NOM). In the presence of EOF the phenanthrene release flux from alginate beads was between 1.4- and 1.8-fold higher than under hydraulic flow conditions with equal bulk water velocity and 30-120-fold higherthan under stagnantwater conditions. Our data suggest that DC-electric fields (0.5-2 V cm(-1)) can stimulate the release of PAH bound to particles exposed to stagnant water zones often found at hydraulic flow regimes restricted by low permeability.

The effect of the nonionic surfactant Brij 35 on the bioavailability of solid and Teflon-sorbed dibenzofuran for Sphingomonas sp. strain HH19k was studied in simple model systems. Growth with dibenzofuran and dibenzofuran-specific oxygen uptake in surfactant-free media and with Brij 35 above the critical micelle concentration (cmc) were compared with dissolution and desorption in the absence of bacteria. Brij 35 accelerated dissolution and biodegradation of solid dibenzofuran by a factor of 2. It also enhanced the initial desorption rate of dibenzofuran from Teflon by this factor. Continuously decreasing desorption rates were attributed to slow diffusion of dibenzofuran inside Teflon, leading to depletion of dibenzofuran in the exterior of the Teflon particles. Surprisingly, Brij 35 slowed the initial biodegradation of desorbing dibenzofuran. We propose two processes that led to low bioavailability of sorbed dibenzofuran in the presence of surfactant. First, desorbing dibenzofuran rapidly accumulated in surfactant micelles, leading to reduced truly water-dissolved dibenzofuran concentration as the factor controlling the biodegradation rate. Second, Brij 35 suppressed the contact between bacteria and Teflon. This increased the average diffusion distance of dibenzofuran to the bacteria, which in turn flattened the gradient of the dissolved dibenzofuran concentration between the sorbent and the cells as the driving force for desorption.