Effect of daptomycin and vancomycin on Staphylococcus epidermidis biofilms: An in vitro assessment using fluorescence in situ hybridization.

Colonization of in-dwelling catheters by microbial biofilms is a major concern in patient health eventually leading to catheter-related blood stream infections. Biofilms are less susceptible to standard antibiotic therapies that are effective against planktonic bacteria. Standard procedure for the detection of microorganisms on the catheter tip is culture. However, viable but non-culturable cells (VBNCs) may be missed. The aim of this study was to evaluate the use of fluorescence in situ hybridization (FISH) as an indicator to visualize and quantify the effect of the antibiotics daptomycin and vancomycin on biofilms in situ. We established an in vitro catheter biofilm model of Staphylococcus epidermidis biofilms on polyurethane catheters. Biofilm activity was measured by FISH and correlated to colony forming units (CFU) data. Digital image analysis was used for quantification of total biofilm mass and the area of the FISH positive biofilm cells. FISH showed a pronounced effect of both antibiotics on the biofilms, with daptomycin having a significantly stronger effect in terms of both reduction of biofilm mass and number of FISH-positive cells. This supports the anti-biofilm capacity of daptomycin. Interestingly, neither antibiotic was able to eradicate all of the FISH-positive cells. In summary, FISH succeeded in visualization, quantification, and localization of antibiotic activity on biofilms. This technique adds a new tool to the arsenal of test systems for anti-biofilm compounds. FISH is a valuable complementary technique to CFU since it can be highly standardized and provides information on biofilm architecture and quantity and localization of survivor cells.

Moraxella catarrhalis induces CEACAM3-Syk-CARD9-dependent activation of human granulocytes.

The human restricted pathogen Moraxella catarrhalis is an important causal agent for exacerbations in chronic obstructive lung disease in adults. In such patients, increased numbers of granulocytes are present in the airways, which correlate with bacteria-induced exacerbations and severity of the disease. Our study investigated whether the interaction of M. catarrhalis with the human granulocyte-specific carcinoembryonic antigen-related cell adhesion molecule (CEACAM)-3 is linked to NF-κB activation, resulting in chemokine production. Granulocytes from healthy donors and NB4 cells were infected with M. catarrhalis in the presence of different inhibitors, blocking antibodies and siRNA. The supernatants were analysed by enzyme-linked immunosorbent assay for chemokines. NF-κB activation was determined using a luciferase reporter gene assay and chromatin-immunoprecipitation. We found evidence that the specific engagement of CEACAM3 by M. catarrhalis ubiquitous surface protein A1 (UspA1) results in the activation of pro-inflammatory events, such as degranulation of neutrophils, ROS production and chemokine secretion. The interaction of UspA1 with CEACAM3 induced the activation of the NF-κB pathway via Syk and the CARD9 pathway and was dependent on the phosphorylation of the CEACAM3 ITAM-like motif. These findings suggest that the CEACAM3 signalling in neutrophils is able to specifically modulate airway inflammation caused by infection with M. catarrhalis.

Response of ammonium removal to growth and transpiration of Juncus effusus during the treatment of artificial sewage in laboratory-scale wetlands.

The correlation between nitrogen removal and the role of the plants in the rhizosphere of constructed wetlands are the subject of continuous discussion, but knowledge is still insufficient. Since the influence of plant growth and physiological activity on ammonium removal has not been well characterized in constructed wetlands so far, this aspect is investigated in more detail in model wetlands under defined laboratory conditions using Juncus effusus for treating an artificial sewage. Growth and physiological activity, such as plant transpiration, have been found to correlate with both the efficiency of ammonium removal within the rhizosphere of J. effusus and the methane formation. The uptake of ammonium by growing plant stocks is within in a range of 45.5%, but under conditions of plant growth stagnation, a further nearly complete removal of the ammonium load points to the likely existence of additional nitrogen removal processes. In this way, a linear correlation between the ammonium concentration inside the rhizosphere and the transpiration of the plant stocks implies that an influence of plant physiological activity on the efficiency of N-removal exists. Furthermore, a linear correlation between methane concentration and plant transpiration has been estimated. The findings indicate a fast response of redox processes to plant activities. Accordingly, not only the influence of plant transpiration activity on the plant-internal convective gas transport, the radial oxygen loss by the plant roots and the efficiency of nitrification within the rhizosphere, but also the nitrogen gas released by phytovolatilization are discussed. The results achieved by using an unplanted control system are different in principle and characterized by a low efficiency of ammonium removal and a high methane enrichment of up to a maximum of 72.7% saturation.

Dynamics of sulphur compounds in horizontal sub-surface flow laboratory-scale constructed wetlands treating artificial sewage.

The knowledge regarding the dynamics of sulphur compounds inside constructed wetlands is still insufficient. Experiments in planted (Juncus effusus) and unplanted horizontal sub-surface-flow laboratory-scale constructed wetlands fed with artificial wastewater were carried out to evaluate the sulphate reduction, the composition and dynamics of generated sulphur compounds, as well as the influence of carbon load and plants on processes of sulphur transformation. In planted and unplanted wetlands, the addition of organic carbon (TOC of about 120 mg L(-1)) immediately affected the transformation of up to 90% of the incoming sulphate (150 mg L(-1)), directing it mainly towards elemental sulphur (30%) and sulphide (8%). During this experimental period, nearly 52% of the transformed sulphate-sulphur was calculated to be immobilized inside the planted wetland and 66% inside the unplanted one. In subsequent experiments, the deficiency of organic carbon inside the planted wetlands favoured the decrease of elemental sulphur in the pore water coupled to retransformation of depot-sulphur to dissolved sulphate. Nearly 90% of the deposited and reduced sulphur was found to be reoxidized. In principle, the results indicate a substantial improvement of this reoxidation of sulphur by oxygen released by the helophytes. Surplus of organic carbon promotes the ongoing sulphate reduction and the stability of deposed and dissolved reduced sulphur compounds. In contrast, inside the unplanted control wetland, a relative stability of the formed sulphur depots and the generated amount of dissolved sulphur compounds including elemental sulphur could be observed independently of the different loading conditions.

Physiological responses of Juncus effusus (Rush) to chromium and relevance for wastewater treatment in constructed wetlands.

Constructed wetlands are increasingly applied for industrial wastewater treatment. However, current knowledge of the stress responses of helophytes to selected toxicants such as dichromate is limited. The goal of the experiments presented here was to characterize the physiological response of Juncus effusus to different concentrations of dichromate dependent upon the growth and constitution of the plants. The growth parameters, shoot length, and dry weight already were strongly affected at low dichromate concentrations of approximately 34 microM. Concentrations of 340 microM impaired chlorophyll fluorescence and a decrease in chlorophyll a started at concentrations higher than 170 microM dichromate. The concentrations of chlorophyll b and carotenoids also were influenced negatively. Thus, the reduction of the pigment content started before any obvious influence on the chlorophyll fluorescence. The highest concentration of dichromate, which caused no permanent inhibition of growth and photosynthesis, was found to be 17 microM K2Cr2O7. In principle, J. effusus is suitable for constructed wetlands to treat chromium-containing wastewater. Because the stress resistance of J. effusus is limited, the maximum concentration of dichromate in the treated wastewater should not exceed 34 microM. The growth parameters, shoot length, and dry weight were sensitive to much lower dichromate concentrations and did react more quickly than the biochemistry-related parameters chlorophyll fluorescence and pigment concentration. Therefore, the example of Juncus effusus shows that the use of only biochemical parameters to define concentration limits for the treatment of dichromate-containing wastewater can lead to incorrect conclusions and result in disturbed long-term operation of the system.

Sulphur transformation and deposition in the rhizosphere of Juncus effusus in a laboratory-scale constructed wetland.

Sulphur cycling and its correlation to removal processes under dynamic redox conditions in the rhizosphere of helophytes in treatment wetlands are poorly understood. Therefore, long-term experiments were performed in laboratory-scale constructed wetlands treating artificial domestic wastewater in order to investigate the dynamics of sulphur compounds, the responses of plants and nitrifying microorganisms under carbon surplus conditions, and the generation of methane. For carbon surplus conditions (carbon:sulphate of 2.8:1) sulphate reduction happened but was repressed, in contrast to unplanted filters mentioned in literature. Doubling the carbon load caused stable and efficient sulphate reduction, rising of pH, increasing enrichment of S(2-) and S(0) in pore water, and finally plant death and inhibition of nitrification by sulphide toxicity. The data show a clear correlation of the occurrence of reduced S-species with decreasing C and N removal performance and plant viability in the experimental constructed wetlands.

Sulphate reduction and the removal of carbon and ammonia in a laboratory-scale constructed wetland.

Sulphate is a normal constituent of domestic wastewater and reduced sulphur compounds are known to be potent inhibitors of plant growth and certain microbial activities. However, the knowledge about sulphate reduction and the effect on the removal of C and N in constructed wetlands is still limited. Investigations in laboratory-scale constructed wetland reactors were performed to evaluate the interrelation of carbon and nitrogen removal with the sulphate reduction by use of artificial domestic wastewater. Carbon removal was found to be only slightly affected and remained at high levels of efficiency (75-90%). Only at sulphate reduction intensities above 75 mgl(-1) (50% removal), a decrease of carbon removal of up to 20% was observed. A highly contrary behaviour of ammonia removal was found in general, which decreased exponentially from 75% to 35% related to a linear increase of sulphate reduction up to 75 mgl(-1) (50% removal). Since sulphate removal is considered to be dependant on the load of electron donors, the carbon load of the system was varied. Variation of the load changed the intensities of sulphate reduction immediately, but did not influence the carbon removal effectiveness. Doubling of the carbon concentration of 200 mgl(-1) BOD(5) for domestic wastewater usually led to sulphate reduction of up to 150 mgl(-1) (100% removal). The findings show that, particularly in constructed wetland systems, the sulphur cycle in the rhizosphere is of high importance for performance of the waste water treatment and may initiate a reconsideration of the amount of sulphate present in the tap water systems.

Anaerobic co-reduction of chromate and nitrate by bacterial cultures of Staphylococcus epidermidis L-02.

Industrial wastewater is often polluted by Cr(VI) compounds, presenting a serious environmental problem. This study addresses the removal of toxic, mutagenic Cr(VI) by means of microbial reduction to Cr(III), which can then be precipitated as oxides or hydroxides and extracted from the aquatic system. A strain of Staphylococcus epidermidis L-02 was isolated from a bacterial consortium used for the remediation of a chromate-contaminated constructed wetland system. This strain reduced Cr(VI) by using pyruvate as an electron donor under anaerobic conditions. The aims of the present study were to investigate the specific rate of Cr(VI) reduction by the strain L-02, the effects of chromate and nitrate (available as electron acceptors) on the strain, and the interference of chromate and nitrate reduction processes. The presence of Cr(VI) decreased the growth rate of the bacterium. Chromate and nitrate reduction did not occur under sterile conditions but was observed during tests with the strain L-02. The presence of nitrate increased both the specific Cr(VI) reduction rate and the cell number. Under denitrifying conditions, Cr(VI) reduction was not inhibited by nitrite, which was produced during nitrate reduction. The average specific rate of chromate reduction reached 4.4 micromol Cr 10(10 )cells(-1 )h(-1), but was only 2.0 micromol Cr 10(10 )cells(-1 )h(-1) at 20 degrees C. The maximum specific rate was as high as 8.8-9.8 micromol Cr 10(10 )cells(-1 )h(-1). The role of nitrate in chromate reduction is discussed.

Influence of the redox condition dynamics on the removal efficiency of a laboratory-scale constructed wetland.

A laboratory reactor planted with Juncus effusus treating an artificial wastewater was used to investigate the short-term and long-term variations and interactions in the redox conditions as well as the removal efficiency of C and the N turnover. The permanent circulation of the process water enabled the micro-gradient processes to be evaluated for an operating period of 20 months. Steady-state conditions were achieved throughout the operating period with high mean removal efficiencies of 92.7% total organic carbon, 82.0% ammonia and 97.6% nitrate. Daily variations in the redox state of the rhizosphere of a few hundred mV were observed, ranging from about -200 to oxidized conditions of about +200 mV and driven by daylight. Variations in pH associated with changes in light and redox were linked to the dynamics of the fates of organic and inorganic carbon species. The ammonia removal processes were found to be firmly established, including for moderately reduced redox conditions with high efficiencies for E(h)>-50 mV. The enrichment of ammonia (up to 13 mg l(-1)) closely linked to the light, particularly during summertime, indicates the existence of hitherto unconsidered additional N turnover pathways in the rhizoplane involving N(2) produced by microbes or released by plants. C turnover was strongly related to the seasonal variation in illumination with minimum efficiencies during the dark season. In addition, it was characterized by oscillation with periods of approximately 1 month. The relationships found are dominant for biofilms on the rhizoplane and decisive for the removal efficiency of especially simple constructed and natural wetlands. The results highlight the importance of helophytes and their physiological specifics for removal processes.

Effects of plants and microorganisms in constructed wetlands for wastewater treatment.

Constructed wetlands are a natural alternative to technical methods of wastewater treatment. However, our understanding of the complex processes caused by the plants, microorganisms, soil matrix and substances in the wastewater, and how they all interact with each other, is still rather incomplete. In this article, a closer look will be taken at the mechanisms of both plants in constructed wetlands and the microorganisms in the root zone which come into play when they remove contaminants from wastewater. The supply of oxygen plays a crucial role in the activity and type of metabolism performed by microorganisms in the root zone. Plants' involvement in the input of oxygen into the root zone, in the uptake of nutrients and in the direct degradation of pollutants as well as the role of microorganisms are all examined in more detail. The ways in which these processes act to treat wastewater are dealt with in the following order: Technological aspects; The effect of root growth on the soil matrix; Gas transport in helophytes and the release of oxygen into the rhizosphere; The uptake of inorganic compounds by plants; The uptake of organic pollutants by plants and their metabolism; The release of carbon compounds by plants; Factors affecting the elimination of pathogenic germs.

Annual cycle of nitrogen removal by a pilot-scale subsurface horizontal flow in a constructed wetland under moderate climate.

The annual course of nitrogen removal in a stable operating subsurface horizontal flow constructed wetland (SSF) in a moderate climate was evaluated using a large pool of data from 4 years of operation. In spring and autumn removal efficiencies were found to depend on the nitrogen load in a linear mode. The efficiencies in winter and summer differed extremely (mean removal rates of 0.15/0.7 g m(-2) d(-1) (11%/53%) in January/August) and were independent of the nitrogen load (0.7-1.7 g m(-2) d(-1)) in principle. Oscillations of the removal rates in spring, forming several maxima, suggest seasonal specific effects caused by the dynamics of the plant-physiology finally determining the nitrification efficiency, i.e. via O(2)-supply. Nitrification is limited by temperature during all seasons and surprisingly in midsummer additionally restricted by other seasonal aspects forming a clear-cut relative nitrification minimum (mean rate of 0.43 g m(-2) d(-1) (32%)) in July. The importance and the effect of the plants' gas exchange and oxygen input into the rhizosphere are discussed. Denitrification was nearly complete in midsummer and was clearly restricted at seasonal temperatures below 15 degrees C.

[The treatment of hydrocephalus in infants and children using hydrostatic valves]. / Die Behandlung des kindlichen Hydrocephalus mit hydrostatischen Ventilen.

Complications of CSF-shunt systems may not only prolong hospitalization, but also influence dramatically the neurological and mental outcome of patients, especially of premature newborns. Shunt complications may be caused by mechanical problems such as shunt occlusion or dysfunction of the valve itself. Another cause of shunt complications is the overdrainage and, lastly, shunt infection. The choice of the specific shunt system seems to play an important role in avoiding the above mentioned complications. In this study 32 children with hydrocephali due to different etiologies were treated with hydrostatic valves, the so called Pädi GAV (pediatric hydrostatic valve) valves developed by Miethke. In addition to mechanical occlusions and shunt infections, the overdrainage related complications, such as subdural hematomas/ fluid collections, slit ventricles and secondary craniosynostoses were taken into consideration. None of the patients showed these complications. On the contrary, postoperative MRI or the ultrasound demonstrated prominent ventricles comparing these with those of non-hydrocephalic children as measured by the Evans-Index and FOR (fronto-to-occipital ratio). Whether the overdrainage-related complications actually reduced using Pädi GAV valves will be shown by further long-term follow ups. This study aims to evaluate the initial experience with the Pädi GAV shunt system, particularly with respect to the overdrainage-related problems.

Model experiments on the microbial removal of chromium from contaminated groundwater.

A bacterial consortium with representatives of sulfate-reducing and denitrifying bacteria was selectively enriched. Model experiments under microaerobic conditions showed that it precipitated chromium from Cr (VI)-containing waters (area of a former electroplating factory, Leipzig, Germany) by two different mechanisms: by sulfate reduction and precipitation as sulfide, and by some direct reduction. Sulfate reduction needed fatty acids as organic substrates and resulted at the first stage in no sulfide accumulation. In the absence of the fatty acids but with straw as organic substrate, the direct reduction of chromium was observed without sulfate reduction. In this case Cr (VI)-reduction rate correlated with that of the denitrification.

Degradation efficiency and molecular size alteration during the aerobic microbial treatment of lignite pyrolysis deposit water.

Investigation into aerobic biological degradation were carried out as part of an extensive programme designed to facilitate the cheap remediation of a pyrolysis waste-water deposit. Attention was focused on the processes of carbon conversion by different populations. The susceptibility of a body of lignite-processing deposit water to microbiological degradation was examined in batch investigations in a Sapromat system and in continuous bench-scale fermenter cultivations, with respect to nutrient supply, inoculation culture and molecular size distribution. It was found that degradation best occurs with an adapted mixed culture. The autochthonous culture removes 30% less dissolved organic carbon (DOC) and has a 40% higher specific oxygen demand. A shortage of phosphorus, investigated with a view to avoiding additional eutrophication problems in the open water in the case of in situ remediation, causes reduced DOC degradation and significantly higher specific oxygen demand. The biological process is overlapped by abiotic oxidation. During aerobic treatment, a concentration of colour-giving aromatic substances of between 0.5 kDa and 5 kDa was observed. This phenomenon is caused by the oxidation of low- and high-molecular-mass compounds. The removal of DOC is limited to 65% and mainly occurs in the range below 0.5 kDa (30%) and in the 0.5-1 kDa range (12%); the removal is negligible in the ranges 1-3 kDa (0.8%) and 3-5 kDa (2%) and a little higher in the ranges 5 kDa-0.3 micron (5%) and above 0.3 micron (6%). In the investigations it was discovered that DOC removal causes in the ranges below 0.5 kDa, 0.5-1 kDa and 5 kDa-0.3 micron mainly as a result of degradation, but the range above 0.3 micron is chiefly caused by bioadsorption. Aerobic microbiological treatment is able to remove most low-molecular-mass substances. In order to remove the macromolecular and colour-giving part of the deposit water, an additional treatment stage, e.g. flocculation, is required.

Spectro-streak picosecond studies of intramolecular charge-transfer fluorescence.

A spectro-streak photometer, an instrument for simultaneously measuring fluorescence intensity, time, and wavelength,I(t, λ), with a single picosecond excitation pulse, has been constructed. Two typical and currently highly topical examples of mesurements are discussed. (1) the temporal development of the fluorescence form the intramolecular charge-transfer (ICT) state of the rigid aromatic compound 4,5-(1'-methylindolino)3,4-naphthanthracene is studied in the protic solvent hexanol. (2) Propyl chain-linked pyrene/N,N-dimethylaniline is used as the model compound to study conformational changes associated with the transition from a contact ion pair to a sandwich exciplex.