Corn silage fungal-based solid-state pretreatment for enhanced biogas production in anaerobic co-digestion with cow manure.

The objective of this research was to use white-rot fungus Trametes versicolor for corn silage pretreatment and to investigate the effect of pretreatment on biogas productivity. Semi-continuous pilot-scale experiment, comprised of two experimental phases, was carried out. In the first phase, operational conditions of the full-scale biogas plant were reproduced at pilot-scale. In that phase, the reactor was daily fed with the mixture of cow manure, digestate from industrial postfermentor, corn grits and ensiled corn silage, and the average methane generation rate was 0.167 m3CH4 kgVS-1. In the second phase, corn grits and ensiled corn silage were replaced with corn silage pretreated with T. versicolor, and the average methane generation rate increased up to 0.236 m3CH4 kgVS-1. The results of this study suggest that application of fungal-based solid-state pretreated corn silage has positive effect on pH stability and increase the biogas productivity.

Biogas Production from Brewer's Yeast Using an Anaerobic Sequencing Batch Reactor.

Renewable energy sources are becoming increasingly important in the beverage and food industries. In the brewing industry, a significant percentage of the used raw materials finishes the process as secondary resource or waste. The research on the anaerobic digestion of brewer's yeast has been scarce until recent years. One of the reasons for this is its use as a secondary resource in the food industry and as cattle feed. Additionally, market value of brewer's yeast is higher than its energy value. Due to the increase of energy prices, brewer's yeast has become of interest as energy substrate despite its difficult degradability in anaerobic conditions. The anaerobic co-digestion of brewer's yeast and anaerobically treated brewery wastewater was studied using a pilot-scale anaerobic sequencing batch reactor (ASBR) seeded with granular biomass. The experiments showed very good and stable operation with an organic loading rate of up to 8.0 kg/(m3·day), and with a maximum achieved organic loading rate of 13.6 kg/(m3·day) in a single cycle. A specific biogas productivity of over 0.430 m3/kg of the total chemical oxygen demand (COD) inserted, and total COD removal efficiencies of over 90% were achieved. This study suggests that the brewer's yeast can be successfully digested in an ASBR without adverse effects on the biogas production from brewer's yeast/wastewater mixtures of up to 8% (by volume). By using the brewer's yeast in the ASBR process, the biogas production from brewery wastewater could be increased by 50%.

Anaerobic Biodegradation of Raw and Pre-treated Brewery Spent Grain Utilizing Solid State Anaerobic Digestion.

The brewery spent grain (BSG) represents approximately 85% of the total quantity of by-products from the brewing industry. The biogas production from the BSG has been the subject of several studies in recent years, due to relatively high energy consumption in the brewing process and due to the increasing energy costs. The biodegradability of raw and pre-treated BSG in a single-stage and two-stage solid-state anaerobic digestion (SS-AD) system was determined in this study. The results show that the BSG have a biogas potential of 120 L/kg(-1). In the single-stage system, the biogas yield obtained from raw BSG (87.4 L/kg(-1)) was almost equal to the yield obtained from the pre-treated BSG (89.1 L/kg(-1)), while the methane yield was 51.9 and 55.3 L/kg(-1) and the biodegradation was 62.0% and 62.2% for raw and pre-treated BSG, respectively. In two-stage SS-AD the pre-treated BSG showed better results, with the biogas yield of 103.2 L/kg(-1) and the biodegradation of 73.6%, while the biogas yield obtained from raw BSG was 89.1 L/kg(-1), with the biodegradation of 63.5%. In two-stage process the obtained methane yields from raw and pre-treated BSG were identical (58.7 L/kg(-1)).

Development and plasticity of mitochondria and electrical properties of the cell membrane in blowfly photoreceptors.

Blowfly photoreceptors are highly energy demanding sensory systems. Their information processing efficiency is enabled by the high temporal resolution of the cell membrane, requiring heavy metabolic support by the mitochondria. We studied the developmental changes of the mitochondrial apparatus and electrical properties of the photoreceptor membrane in the white eyed Calliphora vicina Chalky. Using in vivo microspectrophotometry and Western blot analysis, we found an age-dependent increase in the concentration of mitochondrial pigments. The maximal change occurred during the first week. The age-related changes were smaller in dark-bred than in light-bred flies. The mitochondrial pigment content increased after the switch from dark to light rearing and decreased after the switch from light to dark rearing. The electrical parameters of the photoreceptors were investigated with intracellular recordings. The resting membrane resistance and time constant decreased significantly after eclosion. The decrease was again most significant during the first week of adult life, paralleled with changes in the Na/K pump-dependent hyperpolarizing afterpotential. We conclude that the photoreceptor mitochondria exhibit remarkable ontogenetic and phenotypic plasticity, because the quantity of mitochondrial pigments tightly follows the development of the cell membrane as well as the energy demands of the photoreceptors under different rearing conditions.

The effect of gentian violet on human anterior lens epithelial cells.

PURPOSE: To evaluate whether the gentian violet staining of the anterior lens capsule during the cataract surgery is cytotoxic for the human lens epithelial cells, as an indirect indication of possible toxicity towards the corneal endothelium and the safety of gentian violet application. MATERIALS AND METHODS: Two groups of anterior lens capsules obtained during the cataract surgery, gentian violet stained and non-stained, were incubated with fluorescent dye Fura-2. Their fluorescence, upon excitation at 360 and 380 nm, was imaged to monitor changes in free intracellular calcium concentration ([Ca(2+)]i) in response to pharmacological stimulation by acetylcholine. The [Ca(2+)]i homeostasis is the indicator of cellular function. The changes in [Ca(2+)]i were compared between the two groups. RESULTS: Epithelial cells responded to acetylcholine in both groups of capsules - gentian violet stained (n = 17) and non-stained ones (n = 33). No significant differences of the elicited responses were found in rise time (p = 0.89), decay time (p = 0.61) or amplitude of [Ca(2+)]i (p = 0.96 for 63× and p = 0.26 for 40× objectives) between the two groups of capsules (Student t test). CONCLUSIONS: The staining of the anterior lens capsule with gentian violet during phacoemulsification in concentration of 0.01%, does not have detectable cytotoxic effects, which would affect the [Ca(2+)]i homeostasis in lens epithelial cells. The data, if extrapolated to corneal endothelium, exposed to the same concentration, suggest that gentian violet in concentration of 0.01% is safe as an adjunct for capsule visualization in cataract surgery.

The re-use of Waste-Activated Sludge as part of a "zero-sludge" strategy for wastewater treatments in the pulp and paper industry.

The possibility of introducing the thermo-alkali hydrolysis of Waste-Activated Sludge (WAS) was investigated, in order to enable the use of its solid residue as a raw material in cardboard production and the use of its liquid portion for anaerobic digestion in an UASB reactor. The evaluation of the hydrolysis at pH>12 and T=70°C showed that the microbe cells were disrupted with more than 90% efficiency in less than 2h. The solid portion was hygienised, therefore making it possible to integrate it into the cardboard production as a raw material for less demanding cardboards. Up to 6% addition of the liquid portion of hydrolysed WAS to wastewater decreased the specific biogas production in a pilot-scale UASB from 0.236 to 0.212 m(3)/kg(COD), while the efficiency of the COD removal decreased from 80.4% to 76.5%. These values still guarantee an adequate treatment of the wastewater and an increased biogas production by 16%.

Anaerobic co-digestion of excess brewery yeast in a granular biomass reactor to enhance the production of biomethane.

The anaerobic co-digestion of brewery yeast using granular biomass was studied on the lab, pilot and full-scale. The study shows no adverse effects in the co-digestion of yeast and wastewater in concentrations up to 1.1 (v/v)%. In concentrations up to 2.3% the process is manageable; however, not advisable. In concentrations over 2.8% the process exhibits failure due to the overload with suspended solids. An average specific biogas production of 0.560 m(3)kg(-1) of volatile solids was achieved. Full-scale operation with 0.7% yeast concentration showed a 38.5% increase in the biogas production and a 26.2% increase in the organic loading rate, which resulted in an increase of the biomethane/natural-gas substitute ratio from 10% to 16%. The influence of the yeast addition on the structure of the microbial biomass showed up to 7% dissimilarity in the archaeal and a 32% dissimilarity in the bacterial biomass community, which did not present any difficulties.

Temperature-dependence of Weibel-Palade body exocytosis and cell surface dispersal of von Willebrand factor and its propolypeptide.

BACKGROUND: Weibel-Palade bodies (WPB) are endothelial cell (EC) specific secretory organelles containing Von Willebrand factor (VWF). The temperature-dependence of Ca(2+)-driven WPB exocytosis is not known, although indirect evidence suggests that WPB exocytosis may occur at very low temperatures. Here we quantitatively analyse the temperature-dependence of Ca(2+)-driven WPB exocytosis and release of secreted VWF from the cell surface of ECs using fluorescence microscopy of cultured human ECs containing fluorescent WPBs. PRINCIPAL FINDINGS: Ca(2+)-driven WPB exocytosis occurred at all temperatures studied (7-37°C). The kinetics and extent of WPB exocytosis were strongly temperature-dependent: Delays in exocytosis increased from 0.92 s at 37°C to 134.2 s at 7°C, the maximum rate of WPB fusion decreased from 10.0±2.2 s(-1) (37°C) to 0.80±0.14 s(-1) (7°C) and the fractional extent of degranulation of WPBs in each cell from 67±3% (37°C) to 3.6±1.3% (7°C). A discrepancy was found between the reduction in Ca(2+)-driven VWF secretion and WPB exocytosis at reduced temperature; at 17°C VWF secretion was reduced by 95% but WPB exocytosis by 75-80%. This discrepancy arises because VWF dispersal from sites of WPB exocytosis is largely prevented at low temperature. In contrast VWF-propolypeptide (proregion) dispersal from WPBs, although slowed, was complete within 60-120 s. Novel antibodies to the cleaved and processed proregion were characterised and used to show that secreted proregion more accurately reports the secretion of WPBs at sub-physiological temperatures than assay of VWF itself. CONCLUSIONS: We report the first quantitative analysis of the temperature-dependence of WPB exocytosis. We provide evidence; by comparison of biochemical data for VWF or proregion secretion with direct analysis of WPB exocytosis at reduced temperature, that proregion is a more reliable marker for WPB exocytosis at reduced temperature, where VWF-EC adhesion is increased.

Human anterior lens capsule epithelial cells contraction.

PURPOSE: Human anterior lens epithelial cells, attached to surgically isolated capsules, were found to contract upon stimulation. The purpose of this study was to characterize these contractions, which create gaps between cells, and to assess the underlying physiological mechanisms and their possible association with cataract formation. METHODS: Lens capsules obtained during cataract surgery were stained with fluorescent dye Fura-2. Its fluorescence, upon excitation at 360 and 380 nm, was imaged to monitor changes in cell morphology and cytosolic free Ca(2+) concentrations ([Ca(2+) ](i) ) in response to pharmacological stimulation by acetylcholine (ACh) and to mechanical stimulation by flow of saline or direct contact. RESULTS: Epithelial cells contracted in approximately a third of preparations when stimulated by either ACh application, fluid movement or direct mechanical contact. Contractions started either before or at best simultaneously with the rise in [Ca(2+) ](i). Contractions also occurred when there was hardly any change in [Ca(2+) ](i) upon application of physiological saline alone. The probability of contractions occurring did not differ significantly among cortical, nuclear and combined cortical + nuclear cataract. CONCLUSIONS: This study provides the evidence that contractions of the anterior lens epithelial cells take place in significant portion of human lens anterior capsule postoperative preparations after non-specific stimulation. Contractions are at least partially independent of changes in [Ca(2+) ](i). They can be mechanically induced, are localized and reversible and have a fast response and did not differ among different types of cataract. Physiological and clinical significance of this phenomenon remains to be elucidated.

Changes in redox states of respiratory pigments recorded from the eyes of live blowflies exposed to light stimuli and hypoxia.

Time courses of mitochondrial responses to illumination-induced physiological loads and to hypoxia, were recorded optically from eyes of blowflies Calliphora vicina chalky. We isolated changes in redox states of haems a(3), a, c, and b. Two types of responses to light stimulation were observed. Haems b and a(3) responded with transient oxidation and haems a and c with reduction. The same two groups emerged in response to anoxic exposure. The onset of reduction of haems a and c had virtually no latency, while haems a(3) and b exhibited a transient oxidation followed by reduction only after 10-20 s. The dependence of the steady-state reduction level on [Formula: see text] produced the same groups. Haems a and c were significantly reduced at [Formula: see text] levels around 10 kPa while with haems b and a(3) load-induced oxidation was only replaced by reduction below 2 kPa. We propose haems respond to physiological loads in accordance with their steady-state reduction, which in turn depends largely on barriers for electron transport imposed by the mitochondrial membrane potential. We also propose it may be possible to assess the values of tissue [Formula: see text] and O(2) consumption by monitoring haems that are highly oxidized at rest such as haem a.

Anaerobic digestion of brewery spent grain in a semi-continuous bioreactor: inhibition by phenolic degradation products.

In this study anaerobic digestion of selected lignocellulosic substrate, namely brewery spent grain (BSG), was studied. In order to facilitate anaerobic digestion several types of pretreatment methods were tested such as: mechanical, chemical (alkali and acid) and thermo-chemical. The anaerobic digestion experiments were carried out in a semi-continuous stirred bioreactors with the organic loading rates between 2.9 and 3.9 kgCOD m-3 d-1 (1.9 and 2.5 kgVSS m-3 d-1 respectively) and corresponding hydraulic retention times of 33-39 days. Biogas production and composition, pH, COD, TSS and VSS, short chain fatty acids and phenolic compounds were measured. A significant inhibition of biogas production occurred, depending on the type of substrate pretreatment. There are indications that p-cresol is responsible for process inhibition when its concentration in the reaction mixture exceeds critical value between 115 and 240 mg L-1. Anaerobic digestion of chemically pretreated BSG (acid and alkali) and untreated-raw BSG was inhibited between the days 56 and 63 of the experiment, followed by thermo-chemically pretreated BSG on day 112 and mechanically pretreated BSG on day 126. Analyses of the substrates showed no phenolic compounds either in raw-untreated BSG or pretreated substrates, therefore the recorded p-cresol is an intermediate degradation product, responsible for process inhibition.

Singing greeting card beeper as a finger pulse sensor.

We constructed a robust and low-priced finger pulse sensor from a singing greeting card beeper. The beeper outputs the plethysmographic signal, which is indistinguishable from that of commercial grade sensors. The sensor can be used in school for a number of experiments in human cardiovascular physiology.

Anaerobic digestion of tannery waste: semi-continuous and anaerobic sequencing batch reactor processes.

Disposal of the vast amounts of tannery waste that are currently generated is a significant problem. Anaerobic treatment of different types of tannery waste (fleshings, skin trimmings and wastewater sludge) was investigated. The biochemical methane potential is the same at 37 degrees C or 55 degrees C and an assay of this was shown to be an appropriate screening tool with which to estimate the susceptibility of a substrate to anaerobic digestion. The start-up procedure of a tannery waste thermophilic anaerobic digestion in 100 days using seed from mesophilic digester processing municipal sludge is presented. The specific methane production potential at 55 degrees C is estimated to be 0.617 m(3)kg(-1) of volatile suspended solids for tannery waste sludge, 0.377 m(3)kg(-1) for tannery waste trimmings and 0.649 m(3)kg(-1) for tannery waste fleshings. Additional concerns such as chromium content, salinity and temperature fluctuations were also addressed. Chromium content and salinity showed no adverse effects; however a reactor temperature reduction of 4.4 degrees C led to a drop in biogas production of 25%, indicating a requirement to keep the temperature constant at 55 degrees C.

Multiple mechanisms generate the resting activity of filiform sensilla in the firebug (Pyrrhocoris apterus L.; Heteroptera).

The resting activity was studied in filiform sensilla of the firebug (Pyrrhocoris apterus). Three functional types (T(1), T(2) and T(3)) were detected on the abdomen. A resting discharge of nerve impulses is present in all-always in types T(1) and T(2) and occasionally in type T(3). In T(1) the mean rate is 57, in T(2) 3.3 and in T(3) 0.5 imp/s. Shortening the hair length had a negligible effect on the resting discharge, which indicates an intrinsic origin. The resting activity is highly temperature dependent. In T(1), the activation energy was 56.8, in T(2) 84 and in T(3) 61.4 kJ/mol (Q (10): 2.27, 5.6 and 5.5, respectively). Such values are typical for mechano-transduction, suggesting the involvement of the transduction mechanism itself. The destruction of the hair base in T(1) caused halving of the original discharge rate and shifted the discharge to a regular interval mode. The activation energy decreased to 38 kJ/mol. The destruction of the hair bases in T(2) and T(3) completely abolished the discharge. It appears that at least two mechanisms are involved in the generation of the resting activity in T(1) units while only one can be assumed in case of T(2) and T(3).

Aerobic and two-stage anaerobic-aerobic sludge digestion with pure oxygen and air aeration.

The degradability of excess activated sludge from a wastewater treatment plant was studied. The objective was establishing the degree of degradation using either air or pure oxygen at different temperatures. Sludge treated with pure oxygen was degraded at temperatures from 22 degrees C to 50 degrees C while samples treated with air were degraded between 32 degrees C and 65 degrees C. Using air, sludge is efficiently degraded at 37 degrees C and at 50-55 degrees C. With oxygen, sludge was most effectively degraded at 38 degrees C or at 25-30 degrees C. Two-stage anaerobic-aerobic processes were studied. The first anaerobic stage was always operated for 5 days HRT, and the second stage involved aeration with pure oxygen and an HRT between 5 and 10 days. Under these conditions, there is 53.5% VSS removal and 55.4% COD degradation at 15 days HRT - 5 days anaerobic, 10 days aerobic. Sludge digested with pure oxygen at 25 degrees C in a batch reactor converted 48% of sludge total Kjeldahl nitrogen to nitrate. Addition of an aerobic stage with pure oxygen aeration to the anaerobic digestion enhances ammonium nitrogen removal. In a two-stage anaerobic-aerobic sludge digestion process within 8 days HRT of the aerobic stage, the removal of ammonium nitrogen was 85%.

Treatment of brewery slurry in thermophilic anaerobic sequencing batch reactor.

Treatment of brewery slurry in a thermophilic anaerobic sequencing batch reactor (ASBR) was studied using conventional fully mixed semi-continuous digestion as a control. The process phases were adapted to fit the brewery slurry discharge schedule. ASBR experiments were conducted under different organic loading rates (OLR) from 3.23 to 8.57 kg of COD/m(3)day of reactor and control was conducted with OLR of 3.0 kg of COD/m(3)day. The ASBR COD degradation efficiency was from 79.6% to 88.9%, control experiment efficiency was 65%. ASBR VSS removal efficiency was from 78.5% to 90.5%, control experiment efficiency was 54%. The ASBR methane production yield was from 371 to 418 L/kg COD inserted, control experiment methane yield was 248 L/kg COD inserted. The ASBR process was superior to conventional fully mixed digestion, and is fully adaptable to brewery slurry discharge, needs no additional collection and settling pools and experiences no solids settling problems.

Light dependence of oxygen consumption by blowfly eyes recorded with a magnetic diver balance.

We measured the oxygen (O2) consumption of isolated blowfly eyes using a magnetic diver balance, a device for high-resolution volumetric O2 consumption measurements. The light-induced O2 consumption is at most three times the value of the dark consumption, which is 0.6 nl O2 s(-1) eye(-1), and is in good agreement with the estimates based on electrophysiological data. With longer stimuli the increase follows a double exponential time course. The respective time constants are approximately 2 and 20 s and show no dependence on light intensity, whereas the dependence of amplitudes can be fitted by a Hill equation. Decreasing the stimulus duration reveals that the peak in O2 consumption overshoots the time course induced by long stimuli. We suggest this may be a general feature of mitochondrial activation. The dependence of the O2 consumption peak on stimulus duration at high light intensity has a hump with stimulus durations of 10-20 ms, coinciding with the stimulus durations that start to induce the adaptation of the receptor potential.

A method for dynamic spectrophotometric measurements in vivo using principal component analysis-based spectral deconvolution.

A method was developed for dynamic spectrophotometric measurements in vivo in the presence of non-specific spectral changes due to external disturbances. This method was used to measure changes in mitochondrial respiratory pigment redox states in photoreceptor cells of live, white-eyed mutants of the blowfly Calliphora vicina. The changes were brought about by exchanging the atmosphere around an immobilised animal from air to N2 and back again by a rapid gas exchange system. During an experiment reflectance spectra were measured by a linear CCD array spectrophotometer. This method involves the pre-processing steps of difference spectra calculation and digital filtering in one and two dimensions. These were followed by time-domain principal component analysis (PCA). PCA yielded seven significant time domain principal component vectors and seven corresponding spectral score vectors. In addition, through PCA we also obtained a time course of changes common to all wavelengths-the residual vector, corresponding to non-specific spectral changes due to preparation movement or mitochondrial swelling. In the final step the redox state time courses were obtained by fitting linear combinations of respiratory pigment difference spectra to each of the seven score vectors. The resulting matrix of factors was then multiplied by the matrix of seven principal component vectors to yield the time courses of respiratory pigment redox states. The method can be used, with minor modifications, in many cases of time-resolved optical measurements of multiple overlapping spectral components, especially in situations where non-specific external influences cannot be disregarded.