Investigation on the formation and kinetics of glucose-fed aerobic granular sludge

Aerobic granular sludge was cultivated in a glass sequencing batch reactor (SBR) with glucose synthetic wastewater. The spherical shaped granules were observed on 4th day with the mean diameter of 0.1 mm. With the increase of chemical oxygen demand (COD) concentration of the influent, aerobic granules grew matured, the size of which ranged from 1.2 to 1.9 mm. The aerobic granular sludge could sustain high organic loading rate (about 4.0 g COD L⁻¹ d⁻¹), with good settling ability (settling velocity 36 m/h) and high biomass concentration (MLSS 6.7 ±0.2 g/L). Experimental data indicated that the substrate utilization and biomass growth kinetics followed Monod's kinetics model approximately. The corresponding kinetic coefficients of maximum specific substrate utilization rate (k), half velocity coefficient (K_s), growth yield coefficient (Y) and decay coefficient (K_d) were 13.2 d⁻¹, 275.8 mg/L, 0.183–0.250 mg MLSS/mg COD and 0.023–0.075 d⁻¹, respectively, which made aerobic granules have short setup period, high rate of substrate utilization and little surplus sludge.     

In vivo moment generation and architecture of the human plantar flexors after different shortening–stretch cycles velocities

The purpose of this study was to examine the moment generation of the human plantar flexors and the architecture of the gastrocnemius medialis muscle during and after shortening–stretch cycles in vivo. Fourteen male subjects (30 ± 7 years, 177 ± 7 cm, 80 ± 9 kg) performed a series of electro-stimulated shortening–stretch plantar flexion contractions. The shortening–stretch cycles were performed at three constant angular velocities (25°/s, 50°/s, 100°/s), two amplitudes (15° and 25° ankle angle changes) and at two different stimulation frequencies (30 Hz and 85 Hz). The resultant ankle joint moments were calculated through inverse dynamics. Pennation angle and fascicle length of the m. gastrocnemius medialis at rest and during contractions were measured using ultrasonography. The corresponding ankle moments, kinematics and changes in muscle architecture were analysed at seven time intervals. A three-way analysis of variance (amplitude × velocity × stimulation frequency) and post-hoc test with Bonferroni correction were used to check the amplitude, velocity and stimulation level related effects on moment enhancement (α = 0.05). The results show an ankle joint moment enhancement after shortening–stretch cycles influenced by muscle architectural changes. We found 2–3% isometric ankle joint moment enhancement at steady state, 1.5–2.0 s after the shortening–stretch cycle. However, the observed alteration in muscle architecture after the imposed perturbation, could lead to an underestimation (1–3%) of joint moment enhancement due to the force–length relationship of the triceps surae. Furthermore, the enhancement observed was independent of the shortening–stretch amplitude, velocity and stimulation frequency.     

The seed bank in a subtropical freshwater marsh: implications for wetland restoration

Seed bank samples were collected from Huli Marsh, a subtropical shallow water mountainous marsh in Hunan Province, South China. Core samples were divided into upper and lower layers (each 5 cm in depth) and allowed to germinate in three water levels (0, 5 and 10 cm) over a 4-month period. A total of 51 species germinated and the mean density was 9211 ± 7188 seedlings m⁻². In the top 5 cm 41 species and 5747 ± 5111 seedlings m⁻² germinated, whereas 40 species and 3464 ± 3363 seedlings m⁻² did so from 5–10 cm. Germinated seedling density was significantly higher in the upper layer, largely due to differences in eight species. With increasing experimental water depth, less seedlings germinated: respectively, 9788 ± 7157 m⁻², 2050 ± 2412 m⁻² and 1978 ± 2616 m⁻², of 44, 21 and 19 species, submerged under 0, 5 or 10 cm. Seven species could emerge only in 0 water level. Vallisneria natans occurred only in 5 cm water, whereas Ottelia alismoides occurred in 10 cm water. In the vegetation survey of the marsh, 25 species were recorded, which was less than half of the species recorded in the seed bank. The top 10 dominants in the standing vegetation, accounting for 89% of vegetation abundance, represented only 10% in the seed bank. Twenty germinated species that also occurred in the standing vegetation accounted for 56% of the total seed bank. Our observed number of species germinating from a Chinese wetland seed bank is within the range observed elsewhere in the northern hemisphere (15–113 species).     

Aerobic granulation in sequencing batch reactors with different reactor height/diameter ratios

Effects of reactor height/diameter ratios ranged from 24 to 4 corresponding to reactor settling velocities from 12 to 2 m h^?1 on aerobic granulation were investigated. It was found that granules appeared after 1-week operation and granule volume percentages exceeded 50% after 2–3 weeks in four reactors. In addition, similar granule fraction of 94–96% was found at steady state in all four reactors. Sludge volume index (SVI), average sludge size, biomass density and granule settling velocity at steady state were around 50 ml g^?1, 1800 μm, 53 g l^?1 and 40 m h^?1, respectively, in four reactors. Extracellular polymeric substances (EPS) and specific oxygen uptake rate (SOUR) were around 38 mg g^?1 VSS and 40 mg O₂ g^?1 VSS h^?1, respectively. Denaturing gradient gel electrophoresis (DGGE) fingerprint of sludge in four reactors showed the same microbial population shift during the start-up period and same microbial community structure during steady-state period. These results recommended strongly that reactor height/diameter ratio or reactor setting velocity in the used range in this study did not affect granule formation, physical characteristics, microbial community structure of granules and stable operation of granular sludge reactor. Reactor height/diameter ratio thus can be very flexible in the practice, which is important for the application of aerobic granule technology.     

Sulfate reduction and microbial community of autotrophic biocathode in response to acidity

Microbial electrolysis cells (MECs) with autotrophic biocathode are a promising technology for removal of pollutants in wastewater. The aim of this study was to investigate the effect of initial acidity of wastewater on performance of sulfate-reducing biocathodes. MECs with biocathodes were operated with initial pH values of catholyte ranged from 3.0 to 7.0. The optimum initial pH value was 6.0 with a maximum sulfate reductive rate and biomass of 57 mg L⁻¹ d⁻¹ and 2.1 ± 0.4 mg g⁻¹, respectively. With initial pH 7.0, the pH value of catholyte increased to 9.8 ± 0.2 after an operation cycle, which resulted in low performance of the biocathode. A considerable sulfate reductive rate of 31 ± 0.85 mg L⁻¹ d⁻¹ was achieved with initial pH 3.0. Desulfovibrio sp. grew dominantly with abundance of 46%–66% in the cathode biofilm with initial pH values from 3.0 to 6.0 and contributed to the sulfate reduction. Clostridium and Parapedobacter also had high abundance in pH 6.0 cathode, indicated that interspecies electron transfer between electrochemical active and sulfate-reducing bacteria could play an important role in sulfate removal. The results suggest that acidity of catholyte is an important factor to be considered to utilize autotrophic biocathode MECs for wastewater treatment.     

Regulating effects of climate,net primary productivity,and nitrogen on carbon sequestration rates in temperate wetlands,Northeast China

Temperate wetlands in the Northern Hemisphere have high long-term carbon sequestration rates, and play critical roles in mitigating regional and global atmospheric CO₂ increases at the century timescale. We measured soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP) from 11 typical freshwater wetlands (Heilongjiang Province) and one saline wetland (Jilin Province) in Northeast China, and estimated carbon sequestration rates using ²¹⁰Pb and ¹³⁷Cs dating technology. Effects of climate, net primary productivity, and nutrient availability on carbon sequestration rates (R_carbon) were also evaluated. Chronological results showed that surface soil within the 0–40 cm depth formed during the past 70–205 years. Soil accretion rates ranged from 2.20 to 5.83 mm yr⁻¹, with an average of 3.84 ± 1.25 mm yr⁻¹ (mean ± SD). R_carbon ranged from 61.60 to 318.5 gC m⁻² yr⁻¹ and was significantly different among wetland types. Average R_carbon was 202.7 gC m⁻² yr⁻¹ in the freshwater wetlands and 61.6 gC m⁻² yr⁻¹ in the saline marsh. About 1.04 × 10⁸ tons of carbon was estimated to be captured by temperate wetland soils annually in Heilongjiang Province (in the scope of 45.381–51.085°N, 125.132–132.324°E). Correlation analysis showed little impact of net primary productivity (NPP) and soil nutrient contents on R_carbon, whereas climate, specifically the combined dynamics of temperature and precipitation, was the predominant factor affecting R_carbon. The negative relationship observed between R_carbon and annual mean temperature (T) indicates that warming in Northeast China could reduce R_carbon. Significant positive relationships were observed between annual precipitation (P), the hydrothermal coefficient (defined as P/AT, where AT was accumulative temperature ≥10 °C), and R_carbon, indicating that a cold, humid climate would enhance R_carbon. Current climate change in Northeast China, characterized by warming and drought, may form positive feedbacks with R_carbon in temperate wetlands and accelerate carbon loss from wetland soils.     

Ecological risk assessment of polycyclic aromatic hydrocarbons (PAHs) in the water from a large Chinese lake based on multiple indicators

In the surface water of Lake Chaohu, China, the concentrations of 16 priority polycyclic aromatic hydrocarbons (PAHs) were measured by gas chromatograph–mass spectrometer (GC–MS). Based on the species sensitivity distribution (SSD) model and the probabilistic risk assessment (PRA) model, the indicators were calculated to assess the potential ecological risk of the individual and of multiple congeners of PAHs and their probabilities. The results revealed that the average residual level of the total PAHs (PAH₁₆) in the water ranged from 95.2 to 370.1 ng/L, with a mean value 181.5 ± 70.8 ng/L. The PAH content in the water was dominated by the low-molecular-weight congeners. The multi-substance potentially affected fractions (msPAFs) of the studied PAHs obtained by the SSD model varied from 0.29% (site B3) to 1.58% (site B6), with an average of 0.51 ± 0.34%. The average of the msPAFs (0.93%) for the inflow rivers was greater than that for the western (0.42%) and eastern (0.34%) parts of the lake. The greatest ecological risk probability calculated by the PRA model was found for Pyr (1.55%), followed by Ant (7.07 × 10⁻²%), Fla (2.21 × 10⁻²%), Phe (9.25 × 10⁻⁶%), Nap (1.01 × 10⁻⁵%), Flo (1.16 × 10⁻¹⁴%) and Ace (2.86 × 10⁻¹⁶%). The same order of ecological risks calculated by the two models was found for the studied PAH compounds. The toxicity data might be the primary source of the ecological risk uncertainties, as indicated by the greater values of coefficients of variation (CV) for the toxicity. This study concluded that the combinations of multiple indicators based on the SSD and PRA models for the ecological risk assessment are necessary to provide more general information on the spatial variations and the probabilities of potential ecological risks of the individual and multiple congeners of PAHs.     

Structure and water permeability of fully hydrated diphytanoylPC

Diphytanoylphosphatidylcholine (DPhyPC) is a branched chain lipid often used for model membrane studies, including peptide/lipid interactions, ion channels and lipid rafts. This work reports results of volume measurements, water permeability measurements P_f, X-ray scattering from oriented samples, and X-ray and neutron scattering from unilamellar vesicles at T = 30 °C. We measured the volume/lipid V_L = 1426 ± 1 Å³. The area/lipid was found to be 80.5 ± 1.5 Å² when both X-ray and neutron data were combined with the SDP model analysis (Ku?erka, N., Nagle, J.F., Sachs, J.N., Feller, S.E., Pencer, J., Jackson, A., Katsaras, J., 2008. Lipid bilayer structure determined by the simultaneous analysis of neutron and X-ray scattering data. Biophys. J. 95, 2356–2367); this is substantially larger than the area of DOPC which has the largest area of the common linear chain lipids. P_f was measured to be (7.0 ± 1.0) × 10^?3 cm/s; this is considerably smaller than predicted by the recently proposed 3-slab model (Nagle, J.F., Mathai, J.C., Zeidel, M.L., Tristram-Nagle, S., 2008. Theory of passive permeability through lipid bilayers. J. Gen. Physiol. 131, 77–85). This disagreement can be understood if there is a diminished diffusion coefficient in the hydrocarbon core of DPhyPC and that is supported by previous molecular dynamics simulations (Shinoda, W., Mikami, M., Baba, T., Hato, M., 2004. Molecular dynamics study on the effects of chain branching on the physical properties of lipid bilayers. 2. Permeability. J. Phys. Chem. B 108, 9346–9356). While the DPhyPC head–head thickness (D_HH = 36.4 Å), and Hamaker parameter (H = 4.5 × 10^?21 J) were similar to the linear chain lipid DOPC, the bending modulus (K_C = 5.2 ± 0.5 × 10^?21 J) was 30% smaller. Our results suggest that, from the biophysical perspective, DPhyPC belongs to a different family of lipids than phosphatidylcholines that have linear chain hydrocarbon chains.     

Growth performance and starch utilization in common carp (Cyprinus carpio L.) in response to dietary chromium chloride supplementation

A nutrition trial was conducted on juvenile common carp (Cyprinus carpio), initial mean body weight 15 ± 0.4 g within a controlled facility at 25 ± 0.5 °C. Six diets containing various levels of supplementary Cr (0, 0.2, 0.5, 1.0, 1.5, and 2.0) mg Cr/kg of diet as Cr chloride hexahydrate were fed to carp for a period of 10 weeks. Lower growth performance was observed in fish fed on the control diet and the diet supplemented with the highest level of Cr (2.0 mg Cr/kg). Although fish fed 0.5 mg Cr/kg showed the best growth performance, this was not significantly different (P > 0.05) from fish fed 1.0 mg Cr/kg. The regression of plasma glucose concentration was linear (R² = 0.97 and P value = 0.001) as the Cr content of the diet increased (up to 1.5 mg Cr/kg).Cr carcass content was elevated with an increasing level of dietary Cr supplementation up to 1.5 mg Cr/kg; but fish fed on the diet supplemented with the highest level of Cr (2.0 mg Cr/kg) showed a decrease in Cr carcass content.Histological examination to evaluate the impact of different Cr supplementation on liver and gut tissues showed notable changes. The higher level of Cr (2.0 mg Cr/kg) in the diet gave rise to elevated hepatocyte vacuolization and changes in gut tissue morphology.It appeared that Cr chloride significantly improved growth within a defined range (0.2–1.5) mg Cr/kg without any negative impact, while 2.0 mg Cr/kg in carp diet seems to be the threshold for the initiation of toxicity.     

Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system

Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28 ± 0.15) × 10⁶ U ml⁻¹ and (1.3 ± 0.2) × 10⁶ U ml⁻¹ respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7 ± 0.2) × 10⁶ U ml⁻¹ and (3.5 ± 0.4) × 10⁶ U ml⁻¹ at 2 dpi in Rachiplusia nu larvae and (1.1 ± 0.2) × 10⁶ U ml⁻¹ and (1.0 ± 0.15) × 10⁶ U ml⁻¹ at 5 dpi in Spodoptera frugiperda larvae respectively. R. nu was a better host for FeIFN-α7 and FeIFN-α7xArg expression. The 8xArg tag did not affect the biological activity of FeIFN-α7 and was useful to promote the FeIFN-α7xArg adsorption on ion exchange chromatography (IEC), allowing its purification in a single step from supernatant culture and R. nu larvae. FeIFN-α7xArg was purified from the larval extract with a yield of 70% and a purification factor of 25 free of viruses. We conclude that R. nu larvae are new low-cost hosts for the expression of recombinant FeIFN-α7.     

Partial nitrification under limited dissolved oxygen conditions

Partial nitrification to nitrite is technically feasible and economically favourable, especially when wastewaters contained high ammonium concentrations or low C/N ratios. Partial nitrification can be obtained by selectively inhibiting nitrite-oxidizing bacteria (NOB) through appropriate regulation of the pH, temperature and dissolved oxygen (DO) concentrations. The effect of pH, DO levels and temperature on ammonia oxidation rate and nitrite accumulation was investigated in order to determine the optimal conditions for partial nitrification of synthetic wastewater with high ammonia concentration. The experiments performed at low DO levels to lower the total oxygen needed in the nitrification step, which means great saving in aeration. During the start-up stage pH and DO were set at 7.0–7.4 and 0.5 mg/l, respectively. The reactor was operated until complete partial nitrification was achieved. The effect of pH, DO on partial nitrification was studied, as pH was kept at 6.5, 7.5, 8.5, 9.5 and DO at 0.5±0.2, 1.5±0.2 and 2.5±0.2 mg/l, and temperature at 30 °C. The influence of temperature on k_a value was studied by keeping pH=7.5, DO=1.5 mg/l and temperature was controlled at 12, 20 and 30 °C, respectively. The results showed that partial nitrification to nitrite was steadily obtained and the optimal operational parameters were pH=7.5, DO=1.5 mg/l, T=30 °C based on ammonia oxidation rate and nitrite accumulation rate. The maximum k_a was achieved and to be 115.1×10⁻³ mg NH₄⁺–N (mg VSS h)⁻¹ under this condition.     

Measurement of the influence of titanium hip prosthesis on therapeutic electron beam dose distributions in a novel pelvic phantom

PurposeTo investigate the degree of 18 and 22 MeV electron beam dose perturbations caused by unilateral hip titanium (Ti) prosthesis.MethodsMeasurements were acquired using Gafchromic EBT2 film in a novel pelvic phantom made out of Nylon-12 slices in which a Ti-prosthesis is embedded. Dose perturbations were measured and compared using depth doses for 8 × 8, 10 × 10 and 11 × 11 cm² applicator-defined field sizes at 95 cm source-surface-distance (SSD). Comparisons were also made between film data at 100 cm SSD for a 10 × 10 cm² field and dose calculations made on CMS XiO treatment planning system utilizing the pencil beam algorithm. The extent of dose deviations caused by the Ti prosthesis based on film data was quantified through the dose enhancement factor (DEF), defined as the ratio of the dose influenced by the prosthesis and the unchanged beam.ResultsAt the interface between Nylon-12 and the Ti implant on the prosthesis entrance side, the dose increased to values of 21 ± 1% and 23 ± 1% for 18 and 22 MeV electron beams, respectively. DEFs increased with increasing electron energy and field size, and were found to fall off quickly with distance from the nylon-prosthesis interface. A comparison of film and XiO depth dose data for 18 and 22 MeV gave relative errors of 20% and 25%, respectively.ConclusionThis study outlines the lack of accuracy of the XiO TPS for electron planning in highly heterogeneous media. So a dosimetric error of 20–25% could influence clinical outcome.     

DNA binding acridine–thiazolidinone agents affecting intracellular glutathione

Three new acridine–thiazolidinone derivatives (2a–2c) have been synthesized and their interactions with calf thymus DNA and a number of cell lines (leukemic cells HL-60 and L1210 and human epithelial ovarian cancer cell lines A2780) were studied. The compounds 2a–2c possessed high affinity to calf thymus DNA and their binding constants determined by spectrofluorimetry were in the range of 1.37 × 10⁶–5.89 × 10⁶ M^?1. All of the tested derivatives displayed strong cytotoxic activity in vitro, the highest activity in cytotoxic tests was found for 2c with IC₅₀ = 1.3 ± 0.2 μM (HL-60), 3.1 ± 0.4 μM (L1210), and 7.7 ± 0.5 μM (A2780) after 72 h incubation. The cancer cells accumulated acridine derivatives very fast and the changes of the glutathione level were confirmed. The compounds inhibited proliferation of the cells and induced an arrest of the cell cycle and cell death. Their influence upon cells was associated with their reactivity towards thiols and DNA binding activity.     

Quantification and identification of fungal propagules in well-managed baled grass silage and in normal on-farm produced bales

The viable fungal spora of baled grass silage was quantified and identified. One group of bales (Experiment 1, n = 15) was wrapped in 6 layers of polythene, and was handled and stored for 6 weeks under well-managed conditions. This silage was free of visible fungal contamination after 6 weeks and the film surrounding these bales was visibly undamaged. Moulds were cultured from 9/15 bales, while yeasts were cultured from all bales. A second group of bales (Experiment 2, n = 18) from 9 farms, were wrapped in 4 layers of film, handled and stored for 8 months using normal on-farm procedures. Visible fungal contamination was observed on the surface of most of these bales (15/18) and the film surrounding some bales (8/18) was damaged. In silage sampled from parts of bales that were visually non-mouldy, yeasts were cultured from all bales and moulds from 15/18 bales. Bales in the well-managed group (Experiment 1) had numerically lower numbers of yeasts (mean: 9.7 × 10³ colony-forming units per gram of silage, cfu/g) and lower numbers of moulds (<10¹ cfu/g) compared to the normal on-farm produced group (2.3 × 10⁵ yeast and 1.5 × 10⁵ mould cfu/g; Experiment 2). The most common yeasts in each group of bales were Saccharomyces exiguus (12/15 bales; Experiment 1) and Pichia fermentans (11/18 bales; Experiment 2) and their numbers in all bales ranged from 0 to 10⁵ cfu/g (mean: 8.4 × 10³) and 0 to 1.5 × 10⁶ cfu/g (mean: 1.2 × 10⁵), respectively. Bales contaminated with visible mould growth on their surfaces had higher yeast and mould numbers in visually non-mouldy parts adjacent to the contaminated areas than bales that had no visible mould. Mould numbers were higher (P<0.05) in bales where the polythene film was visibly damaged compared to bales where the polythene film appeared intact. Penicillium roqueforti was not cultured from the well-managed bales, but it was the most common mould in bales prepared using normal on-farm procedures (13/18 bales); propagule numbers in bales ranged from 0 to 7.1 × 10⁵ (mean: 1 × 10⁵ cfu/g). Low numbers of mould propagules, the absence of viable P. roqueforti spores and the absence of mould growth in well-managed bales, emphasises the benefit of applying sufficient film and preventing it from becoming damaged during bale handling and storage.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Chemical compositions and characteristics of organic compounds in propolis from Yemen

Propolis is a gummy material made by honeybees for protecting their hives from bacteria and fungi. The main objective of this study is to determine the chemical compositions and concentrations of organic compounds in the extractable organic matter (EOM) of propolis samples collected from four different regions in Yemen. The propolis samples were extracted with a mixture of dichloromethane and methanol and analyzed by gas chromatography–mass spectrometry (GC–MS). The results showed that the total extract yields ranged from 34% to 67% (mean = 55.5 ± 12.4%). The major compounds were triterpenoids (254 ± 188 mg g⁻¹, mainly α-, β-amyryl and dammaradienyl acetates), n-alkenes (145 ± 89 mg g⁻¹), n-alkanes (65 ± 29 mg g⁻¹), n-alkanoic acids (40 ± 26 mg g⁻¹), long chain wax esters (38 ± 25 mg g⁻¹), n-alkanols (8 ± 3 mg g⁻¹) and methyl n-alkanoates (6 ± 4 mg g⁻¹). The variation in the propolis chemical compositions is apparently related to the different plant sources. The compounds of these propolis samples indicate that they are potential sources of natural bio-active compounds for biological and pharmacological applications.     

High-loading oil palm empty fruit bunch saccharification using cellulases from Trichoderma koningii MF6

Strain Trichoderma koningii D-64 was improved for enhanced cellulase production. A potential mutant MF6 was obtained and its enzymes contained filter paper cellulase (FPase), carboxymethylcellulase (CMCase), β-glucosidase and xylanase with respective activities of 2.0, 1.3, 2.0 and 3.0 folds of those for the parental strain. MF6 cellulases showed enhanced hydrolysis performance for the treated lignocellulosic biomass. Hydrolysis of treated oil palm empty fruit bunch (OPEFB), horticulture wastes (HW) and wood chips (WC) resulted in cellulose to glucose conversion of 96.3 ± 2.2%, 98.2 ± 3.0% and 81.9 ± 1.4%, respectively. The corresponding conversions of xylan to xylose were 96.9 ± 1.5%, 95.0 ± 2.2% and 76.1 ± 3.1%. Consistently, high sugar yield of 770–844 mg/g biomass was obtained for high-loading (10–16%, w/v) of OPEFB hydrolysis and sugar titer of 135.1 g/L was obtained for 16% (w/v) OPEFB loading at 96 h. In addition, MF6 enzymes alone performed equally well for high-loading OPEFB hydrolysis compared to the enzyme mixture of β-glucosidase from Aspergillus niger and cellulase from T. reesei Rut C30.     

An in vitro urinary tract catheter system to investigate biofilm development in catheter-associated urinary tract infections

Biofilm development in urinary tract catheters is an often underestimated problem. However, this form of infection leads to high mortality rates and causes significant costs in health care. Therefore, it is important to analyze these biofilms and establish avoiding strategies. In this study a continuous flow-through system for the cultivation of biofilms under catheter-associated urinary tract infection conditions was established and validated. The in vitro urinary tract catheter system implies the composition of urine (artificial urine medium), the mean volume of urine of adults (1 mL min^–1), the frequently used silicone catheter (foley silicon catheter) as well as the infection with uropathogenic microorganisms like Pseudomonas aeruginosa. Three clinical isolates from urine of catheterized patients were chosen due to their ability to form biofilms, their mobility and their cell surface hydrophobicity. As reference strain P. aeruginosa PA14 has been used. Characteristic parameters as biofilm thickness, specific biofilm growth rate and substrate consumption were observed. Biofilm thicknesses varied from 105 ± 16 μm up to 246 ± 67 μm for the different isolates. The specific biofilm growth rate could be determined with a non invasive optical biomass sensor. This sensor allows online monitoring of the biofilm growth in the progress of the cultivation.     

Macrokinetic determination and water movement in a downward flow biofilter for methanol removal

A steady state model was developed to predict water movement within the biofilter bed. The model’s predictions were compared to experimental data from a downward flow biofilter (50 cm×10 cm i.d.) using compost for removing methanol with concentrations in the range of 0.46–8.41 g m⁻³ and flow rates of 1.36–4.08 m³ per day. The Wani et al. [J. Chem. Technol. Biotechnol. 74 (1999) 9] method of macrokinetic determination was used to estimate the kinetic parameters, and the predicted results showed that this method could be used for methanol removal systems as long as the conversion rate is not limited by diffusion in the biofilm (reaction-controlled regime). The leachate from the biofilter was collected and compared to the model predictions. The amount of collected water increased much more rapidly with inlet methanol concentration than predicted by the model. This shows that there are effects that are not adequately taken into account, such as the breakdown of compost, or biofilm, resulting in loss of water holding capacity, formation of new biofilm, and changes in physical structure. However, this model can be used to estimate the amount of water to be added to ensure that biofilm activity is maximized.     

Differentiating between isolates of Vibrio vulnificus with monoclonal antibodies

Monoclonal antibodies (MAbs) against Vibrio vulnificus (isolate I, VVC and isolate II, VVB) were raised using heat-killed and heat-killed plus SDS–mercaptoethanol treated forms of VVC and VVB for immunizing Swiss mice. Twenty three hybridomas producing MAbs against V. vulnificus were selected and divided into five groups according to their specificities to different V. vulnificus isolates and apparent protein antigens which ranged from ∼ 3–50 kDa. Four groups were specific to V. vulnificus without cross reactivity to either other Vibrio spp. or other bacterial species. In dot blot based assays, one group of MAbs were specific to VVC, with a sensitivity of ∼ 1.6 × 10⁷ CFU ml^− 1 (∼ 1.6 × 10⁴ cells spot^− 1), and bound to proteins of ∼ 50 and ∼ 39 kDa. Other MAbs, binding to proteins ranging from ∼ 3–14 and ∼ 40 kDa, detected VVB (but not VVC) with high sensitivity at ∼ 1.6 × 10⁵ and 4 × 10⁶ CFU ml^− 1 (∼ 1.6 × 10² and 4 × 10³ cells spot^− 1), respectively. In addition, certain MAbs were able to recognize V. vulnificus in tissues by means of immunohistochemistry. The remaining groups demonstrated cross reactivity to Vibrio fluvialis. MAbs from this study can, therefore, detect the difference between some isolates of V. vulnificus and in addition to pathogen detection may, with further antibodies, form the basis of serovar typing isolates in the future.