Effect of Bacillus pumilus ribonuclease on the paramagnetic centers of microbial cells

The potential clinical application of Bacillus pumilus cytotoxic ribonuclease (binase) for selectively inducing the death of tumor cells makes it imperative to investigate its effect on the normal human microflora. Flow cytometry was used to determine that binase concentration causing the apoptosis of cancer cells had no effect of the viability of Escherichia coli K12. The changes in the paramagnetic centers of E. coli K12 cells in the presence of nontoxic binase concentrations revealed by EPR spectroscopy included higher EPR signals from iron-containing proteins (including those from the Fe-S clusters) and of the Mn(II) hyperfine structure. The TMTH spin probe (N-(1-hydroxy-2,2,6,6-tetramethylpiperidine-4-il)-2-methylpropanamide hydrochloride) was used to reveal a twofold increase in the levels of reactive oxygen species (ROS) in the cells, which induced oxidative stress in the enzyme-treated bacteria. Inductively coupled plasma mass spectrometry revealed elevated contents of alkaline (Li, Na, K), alkali earth (Mg, Ca), transition (Cr, Mn, Fe, Cu, Zn), and post-transition metals (Bi, Pb) in the cells. Elevated levels of Cu and Zn (which impair the activity of the respiratory chain enzymes) and of Mn, which is known as a superoxide dismutase cofactor, confirmed development of the oxidative stress in bacteria.     

Effect of gelatin-immobilization on the catalytic activity of enzymes and microbial cells

Summary The immobilization of enzymes and microbial cells within insolubilized gelatin involves both physical entrapment and covalent crosslinking, each one playing its role. The effect of this dual type of bonding on the kinetic parameters and activity yield of three enzymes (acid phosphatase, invertase and -glucosidase) and of whole microbial cells belonging to three yeast species (Saccharomyces cerevisiae,Candida utilis andKluyveromyces marxianus) have been investigated.     

Effect of separator and inoculum type on electricity generation and microbial community in single-chamber microbial fuel cells

Single-chamber microbial fuel cell (SMFC)-I consisted of 4 separator-electrode assemblies (SEAs) with two types of cation exchange membrane (CEM: Nafion and CMI 7000) and an anion exchange membrane (AEM: AMI 7001). SMFC-II consisted of 4 SEAs with Nafion and three types of nonwoven fabric. SMFC-I and -II were inoculated with anaerobic digested and activated sludge, respectively, and operated under fed-batch mode. In SMFC I, AEM-SEA showed a maximum power density (PD_max). Nafion-SEA showed a PD_max in SMFC II, which was similar to that of Nafion–SEA of SMFC I. Although different bacteria were developed in SMFC-I (Deltaproteobacteria and Firmicutes) and SMFC-II (Gammaproteobacteria, Betaproteobacteria and Bacteroidetes), the inoculum type little affects electricity generation. Variations of pH and oxygen in biofilm have influenced microbial community structure and electricity generation according to the electrode and separator material. Although the electricity generation of non-woven fabric-SEA was less than that of Nafion-SEA, the use of non-woven fabrics is expected to reduce the construction and operating costs of MFCs.     

Effect of ammonium and nitrate on current generation using dual-cathode microbial fuel cells

These studies were conducted to determine the effects of various concentrations of ammonium and nitrate on current generation using dual-cathode microbial fuel cells (MFCs). Current generation was not affected by ammonium up to 51.8+/-0.0 mg/l, whereas 103.5+/-0.0 mg/l ammonium chloride reduced the current slightly. On the other hand, when 60.0+/-0.0 and 123.3+/-0.1 mg/l nitrate were supplied, the current was decreased from 10.23+/-0.07 mA to 3.20+/-0.24 and 0.20+/-0.01 mA, respectively. Nitrate did not seem to serve as a fuel for current generation in these studies. At this time, COD and nitrate removal were increased except at 123+/-0.1 mg NO(3)(-)/l. These results show that proper management of ammonium and nitrate is very important for increasing the current in a microbial fuel cell.     

Bioelectricity production from acidic food waste leachate using microbial fuel cells: Effect of microbial inocula

The effects of three different inocula (domestic wastewater, activated sludge, and anaerobic sludge) on the treatment of acidic food waste leachate in microbial fuel cells (MFCs) were evaluated. A food waste leachate (pH 4.76; 1000 mg chemical oxygen demand (COD)/L) was used as the substrate. The results indicate that the leachate itself can enable electricity production in an MFC, but the co-addition of different inocula significantly reduces the start-up time (approximately 7 days). High COD and volatile fatty acids removal (>87%) were obtained in all MFCs but with only low coulombic efficiencies (CEs) (14–20%). The highest power (432 mW/m³) and CE (20%) were obtained with anaerobic sludge as the co-inoculum. Microbial community analysis (PCR-DGGE) of the established biofilms suggested that the superior performance of the anaerobic sludge-MFC was associated with the enrichment of both fermentative (Clostridium sp. and Bacteroides sp.) and electrogenic bacteria (Magnetospirillum sp. and Geobacter sp.) at the anode.     

Effect of some factors on determination of viable microbial cells by peroxyoxalate chemiluminescence method

The effects of physical and chemical factors on the production of H₂O₂ from Escherichia coli cells were studied. When 20 mmol 1^-1 Tris-HCl buffer was used for this purpose the electron transport system (ETS) showed the highest activity at pH 7.6-8.2. KCN promoted the production of H₂O₂ from E. coli cells, and the optimum concentration was changed in different reaction times and pH values. Glucose, 5 mg ml^-1, increased the ETS activity about twofold. The other substrates and surfactants did not increase the chemiluminescence intensity. NaNO₂ and Na₂SO₄ in inorganic salts significantly reduced the ETS activity above 70%. In addition, the optimum temperature for the production of H₂O₂ was 30°C in this study. When glucose (5 mg ml^-1) and KCN (0.2 mmol 1^-1) were added to the reaction buffer containing 0.5 mmol 1^-1 menadione, the detectable minimum cell densities (averages of triplicate assay) of E. coli, Enterobacter cloacae and Serratia marcescens were 5 times 10³ cells ml^-1, 10⁴ cells ml^-1 and 10⁴ cells ml^-1 respectively.     

Effect of electrode surface properties on enhanced electron transfer activity in microbial fuel cells

The influence of electrode surface chemistry over biofilm growth was evaluated for photo‐bioelectrocatalytic fuel cell. A consortium of photosynthetic bacteria was grown onto different electrodes designed with polyethylenimine (PEI) and multiwall carbon nanotubes as hydrophilic and hydrophobic modifier, respectively. The designed electrodes were loaded with 0.08, 0.17, and 0.33 μg/cm² of PEI to change the hydrophilicity. However, 0.56, 0.72, and 0.83 mg/cm² of multiwall carbon nanotubes were used to alter the hydrophobicity of the electrodes. The surface chemistry of electrode and bio‐interaction was evaluated as a function of contact angle and biofilm formation. The results were compared with those obtained with a carbon paper electrode. The contact angle on the untreated electrode (carbon paper) was 118°, whereas for hydrophobic and hydrophilic electrodes, the maximum and minimum contact angles were 170° and 0°, respectively. Interestingly, the maximum biofilm growth (0.2275 g, wet basis) was observed on highly hydrophobic surface; however, the maximum electrochemical performance (246 mV) was shown by the most hydrophilic electrode surface. PEI‐based electrode with good biofilm formation showed comparatively higher electrogenic activity.     

Effect of external resistance on bacterial diversity and metabolism in cellulose-fed microbial fuel cells

External resistance affects the performance of microbial fuel cells (MFCs) by controlling the flow of electrons from the anode to the cathode. The purpose of this study was to determine the effect of external resistance on bacterial diversity and metabolism in MFCs. Four external resistances (20, 249, 480, and 1000 Ω) were tested by operating parallel MFCs independently at constant circuit loads for 10 weeks. A maximum power density of 66 mW m⁻² was achieved by the 20 Ω MFCs, while the MFCs with 249, 480, and 1000 Ω external resistances produced 57.5, 27, and 47 mW m⁻², respectively. Denaturing gradient gel electrophoresis analysis of partial 16S rRNA genes showed clear differences between the planktonic and anode-attached populations at various external resistances. Concentrations of short chain fatty acids were higher in MFCs with larger circuit loads, suggesting that fermentative metabolism dominated over anaerobic respiration using the anode as the final electron acceptor.     

Microencapsulation of microbial cells

The high level of biocatalysts such as microbial cells and enzymes plays an important role in increasing the productivity of a bioreactor. The beads entrapped with microbial cells are not strong enough for long-term use. The small void space of polymer matrix and the leakage of cells limit a final cell loading in the beads. The recent success of encapsulating microbial cells makes it possible to prepare dense biocatalyst composed of recombinant microbial cells. In addition to encapsulating microbial cells, immobilization of animal and plant cells in capsules is also briefly described.     

Effect of initial carbon sources on the performance of microbial fuel cells containing Proteus vulgaris

Mediator-coupled microbial fuel cells containing Proteus vulgaris were constructed and the cell performance was tested. Fuel cell efficiency depended on the carbon source in the initial medium of the microorganism. Maltose and trehalose were not utilized substantially by P. vulgaris; however, their presence in the initial medium resulted in enhanced cell performance. In particular, galactose showed 63% coulombic efficiency in a biofuel cell after P. vulgaris was cultured in a trehalose-containing medium. This work demonstrates that optimum utilization of carbon sources by microorganisms, which leads to the maximization of fuel cell performance, is possible simply by adjusting initial carbon sources.     

Characterization of helper T cells induced by the type 2 antigen polyvinylpyrrolidone

Type 2 antigens are usually unable to prime for IgG memory responses or to activate helper T cells (TH) necessary for memory B cell generation. Previous studies from this laboratory have established that low doses (0.0025 microgram) of polyvinylpyrrolidone (PVP) or a T-dependent form of PVP, PVP-coupled horse red blood cells (PVP-HRBC) can activate PVP-specific TH. The present study was undertaken in order to determine some of the characteristics of the TH activated by PVP and to compare their properties with those of classical TH1 and of TH2 cells described in many T-dependent systems. TH activated with either 0.0025 microgram of PVP or PVP-HRBC were characterized with respect to cell surface antigens, Igh restriction and generation in mice expressing an X-linked immune defect (xid mice). PVP-specific TH are similar to TH1 cells in that they are required for the production of IgG subclasses absent in primary responses and have the Lyt-1+, L3T4+, I-J-surface phenotype. These TH may not be identical with TH1 cells, however, since they are I-A+ and Igh restricted. PVP-specific TH can be generated in xid mice which do not produce antibody in a primary anti-PVP response and do not develop a memory response to PVP, regardless of whether it is presented as a type 2 or T-dependent antigen.     

Characterization of suppressor T cells induced with the thymus-independent antigen polyvinylpyrrolidone coupled to syngeneic cells

CAF₁ mice injected iv with polyvinylpyrrolidone (PVP) coupled to syngeneic spleen cells (PVP-SC) and challenged several days later with 0.25 μg PVP produced fewer PVP-specific IgM plague-forming cells (PFC) than mice injected with Mock-SC. Both 10,000 and 360,000 MW PVP could induce unresponsiveness after coupling to SC. The unresponsiveness induced by PVP-SC was shown to be mediated, at least in part, by antigen-specific suppressor T cells (T_S). The PVP-specific T_S were I-J positive and belonged to the Lyt 1⁺ 2⁺ subset of T cells. The T_s precursors were sensitive to 20 mg/kg cyclophosphamide (Cy) and to antilymphocyte serum (ALS). Kinetics studies suggested that unresponsiveness induced by PVP-SC may be of two types since unresponsiveness in the intact animal appeared earlier and did not last as long as detectable T_S activity.     

Effect of humic acids on electricity generation integrated with xylose degradation in microbial fuel cells

Pentose and humic acids (HA) are the main components of hydrolysates, the liquid fraction produced during thermohydrolysis of lignocellulosic material. Electricity generation integrated with xylose (typical pentose) degradation as well as the effect of HA on electricity production in microbial fuel cells (MFCs) was examined. Without HA addition the maximum power density increased from 39.5 mW/m(2) to 83 mW/m(2) when initial xylose concentrations increased from 1.5 to 30 mM, while coulombic efficiency ranged from 13.5% to 52.4% for xylose concentrations of 15 and 0.5 mM, respectively. Compared to controls where HAs were not added, addition of commercial HA resulted in increase of power density and coulombic efficiency, which ranged from 7.5% to 67.4% and 24% to 92.6%, respectively. Digested manure wastewater (DMW) was tested as potential mediator for power generation due to its content of natural HA, and although it could produce higher coulombic efficiency namely 32.2% than the control of 18.3%, showed lower power density which was approx. 57 mW/m(2) in comparison to power density of the control which was 69 mW/m(2). Presence of commercial HA or DMW in the anode chamber resulted in faster xylose degradation and formation of more oxidized products (acetate and formate) as well as less reduced products (lactate and ethanol) compared to the controls. The reduced power generation in the presence of DMW was attributed to the presence of bacterial inhibitors such as phenolic compounds. Therefore, new feedstocks for MFCs, containing both mediators and substrates, such as lignocellulose hydrolysates should be considered for their applicability in MFCs.     

Effect of static magnetic field on electricity production and wastewater treatment in microbial fuel cells

The effect of a magnetic field (MF) on electricity production and wastewater treatment in two-chamber microbial fuel cells (MFCs) has been investigated. Electricity production capacity could be improved by the application of a low-intensity static MF. When a MF of 50 mT was applied to MFCs, the maximum voltage, total phosphorus (TP) removal efficiency, and chemical oxygen demand (COD) removal efficiency increased from 523?±?2 to 553?±?2 mV, ～93 to ～96 %, and ～80 to >90 %, respectively, while the start-up time and coulombic efficiency decreased from 16 to 10 days and ～50 to ～43 %, respectively. The MF effects were immediate, reversible, and not long lasting, and negative effects on electricity generation and COD removal seemed to occur after the MF was removed. The start-up and voltage output were less affected by the MF direction. Nitrogen compounds in magnetic MFCs were nitrified more thoroughly; furthermore, a higher proportion of electrochemically inactive microorganisms were found in magnetic systems. TP was effectively removed by the co-effects of microbe absorption and chemical precipitation. Chemical precipitates were analyzed by a scanning electron microscope capable of energy-dispersive spectroscopy (SEM-EDS) to be a mixture of phosphate, carbonate, and hydroxyl compounds.