Support media for microbial adhesion in an anaerobic fluidized-bed reactor

The most significant variable in anaerobic digestion in an anaerobic fluidized-bed reactor (AFBR) is the selection of the support medium for microbial adhesion. Using eight kinds of media, cristobalite, zeolite, vermiculite, granular active carbon, granulated clay, pottery stone, volcanic ash, and slag, we examined the physical properties of each medium, microbial adhesion, loading rates of organic matter and removal efficiencies in an AFBR. It appeared that good performance as a support medium was associated with rougher surfaces rather than with larger surface areas, because, although cristobalite had a much smaller surface area (50 m²/g) than that of the granular active carbon (1,125 m²/g), it had a very rough surface with many tubercular processes, by which a maximum loading rate of TOC of 8 g/l·d could be achieved in a synthetic wastewater. Moreover, it appeared to be important that the surface of the medium has a positive charge judging from the difference in performance between cristobalite and zeolite. That is, the two media were charged positively and negatively at pH 7, respectively. As a result, microorganisms, charged negatively in general, could adhere more easily to cristobalite than to zeolite, which was confirmed under a scanning electron microscope (SEM) and by amounts of microbial cells adhering (85 mg cells/g). The upflow linear velocity to allow twice the expanded volume (defined as the ratio of the expanded height to the static height) was decreased to half (0.13 cm/s) by microbial adhesion. In conclusion, a suitable medium for adherence of microorganisms in AFBR should have a rough and positively charged surface rather than a large surface area.     

Anaerobic treatment of wastewater with high salt content from a pickled-plum manufacturing process

Anaerobic treatment of wastewater with a high salt content generated during a pickled-plum manufacturing process (TOC, 14g/l; ash, 150g/l; pH 2.7, hereafter called pickled-plum effluent) was investigated for its effect on the high salt content of the wastewater. The synthetic wastewater, including NaCl up to 30g/l, was treated anaerobically by a draw and fill method (treatment temperature, 37°C; volumetric loading rate of organic matter, 2g/l·d). The TOC removal efficiency and rate of gas evolution then gradually decreased as salt content increased, although stable operation was maintained. At NaCl concentrations above 30g/l, TOC removal efficiency decreased rapidly and stable operation could not be maintained. Five-fold-diluted pickled-plum effluent was treated by the same method at a volumetric TOC loading rate of 2.9g/l·d with a TOC removal efficiency of 71%. Five-fold-diluted pickled-plum effluent was also treated in an anaerobic fluidized-bed reactor (AFBR) at a maximum volumetric TOC loading rate of 3.0g/l·d, which gave almost the same results as the draw and fill method. However, ten-fold-diluted pickled-plum effluent could be treated in the AFBR at a maximum volumetric TOC loading rate of 11.1g/l·d with a TOC removal efficiency of 84.6%. The red pigment in the pickled-plum effluent was completely decolorized by the anaerobic treatment.     

木质纤维素燃料乙醇蒸馏废水的高温厌氧处理及菌群结构分析

【目的】为开发高效的高浓度木质纤维素燃料乙醇蒸馏废水厌氧处理及资源化利用工艺,以活性炭为载体,在实验室规模上对高温厌氧流化床反应器处理木质纤维素燃料乙醇蒸馏废水进行研究。【方法】反应器经65 d梯度驯化后启动,对工艺参数进行一系列优化,并通过基于16S rRNA基因的分子生态学技术分析厌氧污泥中的优势菌群。【结果】实验获得了最优的反应条件和处理效果:厌氧流化床反应器(Anaerobic fluidized bed reactor,AFBR)在温度55±1°C、有机负荷率(OLR)13.8 g COD/(L·d)及水力停留时间(HRT)48 h操作时,COD去除率达到90%以上,同时甲烷产率达到290 mL/g COD;菌群鉴定分析结果显示高温厌氧活性污泥中Clostridia所占比例最大,产甲烷菌属以Methanoculleus和Methanosarcina为主,其它功能菌群主要为Alphaproteobacteria等。【结论】AFBR反应器可高效降解木质纤维素燃料乙醇蒸馏废水并产生生物能源甲烷,其反应体系内微生物种类丰富。     

Anaerobic treatment of wastewater from a food-manufacturing plant with a low concentration of organic matter and regeneration of usable pure water

Wastewater from a food-manufacturing plant with a low concentration of organic matter below 100 mg/l TOC was first treated at 37°C in an anaerobic fluidized-bed reactor (AFBR) or in an upflow anaerobic sludge blanket (UASB). The TOC removal efficiency in both reactors decreased from 85% to 65% as the influent TOC concentration decreased from 100 to 35 mg/l at a hydraulic retention time (HRT) of 6 h. Treatment at an HRT of 4 h resulted in an effluent TOC concentration of 11 to 15 mg/l. The concentration of suspended solids in the effluent could be reduced to 20 mg/l, which corresponded to 7% of that of the influent. The effluent from both reactors was then treated anaerobically in a fixed-bed reactor system. The TOC concentration and optical density (OD) of the effluent from the aerobic treatment were reduced to 5 mg/l and 0.005, respectively, at an HRT of 2 h. When anaerobically or aerobically treated effluent was passed over an activated carbon column, the effluent TOC concentration was reduced to 2 to 3 mg/l. The conductivity of 1.3 mS/cm in raw wastewater, which was not removed through the above treatments, was reduced to 0.001 mS/cm on an ion-exchange resin column. An effluent quality corresponding to that of ultra-pure water for industrial use was finally attained by the treatment in this multi-step system.     

Individual and coupled effects of echinoderm extracts and surface hydrophobicity on spore settlement and germination in the brown alga Hincksia irregularis

Marine substrata possess cues that influence the behavior of fouling organisms. Initial adhesion of fouling algal zoospores to surfaces is also theorized to depend primarily upon interactions between substrata and spore cell bodies and flagellar membranes. In an effort to identify cues and surface characteristics that influence spore settlement and early development, the effects of bioactive echinoderm extracts, surface charge, and surface hydrophobicity were examined individually and in tandem on zoospore settlement and germination in Hincksia irregularis. Experiments utilizing 96-well plastic culture plates confirmed that spore settlement and germination were significantly affected by surface charge and hydrophobicity as well as by echinoderm metabolites, both individually and in tandem. Spore settlement rates in the dark over 30 min were > 400% higher on hydrophobic surfaces than on positively and negatively charged surfaces. Spore germling numbers were > 300% higher on hydrophobic surfaces than on positively and negatively charged surfaces when spores were allowed to settle in the light for 30 min and the settled spores allowed to subsequently germinate for 24 h. Spore germling numbers were consistently > 25% higher on hydrophobic surfaces than on positively and negatively charged surfaces when equal numbers of spores were allowed to completely settle in the light and subsequently germinate for 24 h. H. irregularis germ tube lengths were also significantly longer on positively charged plates than on negatively charged plates. All echinoderm extracts tested had significant effects on germination and settlement at levels below those of estimated ecological concentrations. Short-term (30 min) exposure and subsequent germination experiments indicated that higher concentrations of extracts had rapid toxic effects on algal spores. Synchronous effects of echinoderm extracts and plate charge upon spore settlement varied considerably and did not show a strong dose response relationship. Long-term (24 h) exposure of spores to echinoderm extracts had dosage dependent effects on germination and spore survival. The results of this study indicate that H. irregularis spores possess the capacity for complex responses to their environment, utilizing combined cues of surface charge, surface energy and biochemistry to determine where to settle and germinate. These responses may aid spores in the detection of suitable substrata and conditions for settlement in the marine environment.     

Cell proliferation and cell sheet detachment from the positively and negatively charged nanocomposite hydrogels

The charged nanocomposite hydrogels (NC gels) were synthesized by copolymerization of positively or negatively chargeable monomer with N‐isopropylacrylamide (NIPAm) in the aqueous suspension of hectorite clay. The ionic NC gels preserved the thermo‐responsibility with the phase‐transition temperature below 37°C. The L929 cell proliferation was sensitive to charge polarity and charge density. As compared to the PNIPAm NC gel, the cationic NC gels with <5 mol % of 2‐(dimethylamino)ethyl methacrylate (DMAEMA) showed improved cell proliferation, whereas the cells grew slowly on the gels with negatively charged 2‐acrylamido‐2‐methylpropane sulfonic acid (AMPSNa). By lowering temperature, rapid cell sheet detachment was observed from the surface of ionic NC gels with 1 mol % of ionizable monomers. However, lager amount of AMPSNa or DMAEMA did not support rapid cell sheet detachment, probably owing to the adverse swelling effects and/or enhanced electrostatic attraction. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 58–65, 2014.     

Individual and Coupled Effects of Echinoderm Extracts and Surface Hydrophobicity on Spore Settlement and Germination in the Brown Alga Hincksia irregularis

Marine substrata possess cues that influence the behavior of fouling organisms. Initial adhesion of fouling algal zoospores to surfaces is also theorized to depend primarily upon interactions between substrata and spore cell bodies and flagellar membranes. In an effort to identify cues and surface characteristics that influence spore settlement and early development, the effects of bioactive echinoderm extracts, surface charge, and surface hydrophobicity were examined individually and in tandem on zoospore settlement and germination in Hincksia irregularis. Experiments utilizing 96-well plastic culture plates confirmed that spore settlement and germination were significantly affected by surface charge and hydrophobicity as well as by echinoderm metabolites, both individually and in tandem. Spore settlement rates in the dark over 30?min were >?400% higher on hydrophobic surfaces than on positively and negatively charged surfaces. Spore germling numbers were >?300% higher on hydrophobic surfaces than on positively and negatively charged surfaces when spores were allowed to settle in the light for 30?min and the settled spores allowed to subsequently germinate for 24?h. Spore germling numbers were consistently >?25% higher on hydrophobic surfaces than on positively and negatively charged surfaces when equal numbers of spores were allowed to completely settle in the light and subsequently germinate for 24?h. H. irregularis germ tube lengths were also significantly longer on positively charged plates than on negatively charged plates. All echinoderm extracts tested had significant effects on germination and settlement at levels below those of estimated ecological concentrations. Short-term (30?min) exposure and subsequent germination experiments indicated that higher concentrations of extracts had rapid toxic effects on algal spores. Synchronous effects of echinoderm extracts and plate charge upon spore settlement varied considerably and did not show a strong dose response relationship. Long-term (24?h) exposure of spores to echinoderm extracts had dosage dependent effects on germination and spore survival. The results of this study indicate that H. irregularis spores possess the capacity for complex responses to their environment, utilizing combined cues of surface charge, surface energy and biochemistry to determine where to settle and germinate. These responses may aid spores in the detection of suitable substrata and conditions for settlement in the marine environment.     

Liquefaction and gasification during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction

In order to improve the gas evolution rate during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction, the liquefaction and gasification processes were separately investigated. In the liquefaction process (including the acidification process), treatment at a pH above 6 had no major effects on the liquefaction and acidification rates. However, the VFA production rates were 880 and 320 mg/l·d during mesophilic (37°C) and thermophilic (53°C) liquefaction, respectively. Mesophilic conditions were superior to thermophilic conditions in the liquefaction. With respect to the gasification process, a high TOC volumetric loading rate of 21 g/l·d was achieved during thermophilic gasification. However, the mesophilic gasification did not yield stable data, even at a low TOC volumetric loading rate of 2 g/l·d. The gas yield was 1.7 l/g TOC consumed during thermophilic gasification. The thermophilic liquefaction and thermophilic gasification reactors were connected in series and a two-phase experiment was conducted with the reactors at various volumetric ratios. The maximum gas evolution rate of 1.43 l/l·d was achieved with a combination of a gasification reactor with a 0.45l working volume and liquefaction reactor with a 2l working volume. This rate was 1.7 times higher than the rate obtained in a previous study.     

Treatment of coffee waste by slurry-state anaerobic digestion

Slurries containing 20% (w/v) coffee waste solids were treated anaerobically in one- and two-phase thermophilic methane fermentation systems (53°C) with or without pH control. In one-phase methane fermentation using a roller bottle reactor, the maximum gas evolution rate of 0.87 l/l·d was achieved during treatment for 91 d. However, this one-phase methane fermentation did not yield reproducible data. In a two-phase methane fermentation system consisting of a completely stirred tank reactor type (CSTR-type) liquefaction reactor without pH control and an anaerobic fluidized bed type gasification reactor, three-repetitions of treatment were conducted. Each treatment was very stable and the average gas evolution rate per volume of the gasification reactor was about 2.4 l/l·d. Two-repetitions of treatment were then done while controlling pH in the liquefaction at more than 6. The average gas evolution rate per volume of gasification reactor was found to have increased to 10.2 l/l·d, a value which corresponded to 0.84 l/l·d per total volume, including the liquefaction reactor. It was observed that treatment in a two-phase methane fermentation could be repeated in a stable fashion even in the closed system without discharging anything but the coffee waste residues.     

Dry anaerobic ammonia–methane production from chicken manure

The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65°C while quite low yields of 8.45 and 6.34 ml g⁻¹ VS (volatile solids) were observed at 35 and 45°C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65°C for 8 days: the stripping for 1 day at 85°C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55°C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g⁻¹ VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g⁻¹ VS was achieved during the second batch for 55 days.     

Adsorption of viruses to charge-modified silica.

The purpose of this study was to provide a clearer understanding of virus adsorption, focusing specifically on the role of electrostatic interactions between virus particles and adsorbent surfaces. The adsorption of poliovirus 1, reovirus types 1 and 3, and coliphages MS-2 and T2 to colloidal silica synthetically modified to carry either positive or negative surface charge was evaluated. Adsorption experiments were performed by combining virus and silica in 0.1-ionic-strength buffers of pH 4.0, 6.4, and 8.5. Samples agitated for specified adsorption periods were centrifuged to pellet adsorbent particles plus adsorbed virus, and the supernatants were assayed for unadsorbed virus. All viruses adsorbed exclusively to negatively charged silica at pH values below their isoelectric points, i.e., under conditions favoring a positive surface charge on the virions. Conversely, all viruses adsorbed exclusively to positively charged silica at pH values above their isoelectric points, i.e., where virus surface charge is negative. Viruses in near-isoelectric state adsorbed to all types of silica, albeit to a lesser degree.     

Adsorption of viruses to charge-modified silica

The purpose of this study was to provide a clearer understanding of virus adsorption, focusing specifically on the role of electrostatic interactions between virus particles and adsorbent surfaces. The adsorption of poliovirus 1, reovirus types 1 and 3, and coliphages MS-2 and T2 to colloidal silica synthetically modified to carry either positive or negative surface charge was evaluated. Adsorption experiments were performed by combining virus and silica in 0.1-ionic-strength buffers of pH 4.0, 6.4, and 8.5. Samples agitated for specified adsorption periods were centrifuged to pellet adsorbent particles plus adsorbed virus, and the supernatants were assayed for unadsorbed virus. All viruses adsorbed exclusively to negatively charged silica at pH values below their isoelectric points, i.e., under conditions favoring a positive surface charge on the virions. Conversely, all viruses adsorbed exclusively to positively charged silica at pH values above their isoelectric points, i.e., where virus surface charge is negative. Viruses in near-isoelectric state adsorbed to all types of silica, albeit to a lesser degree.     

Temperature effects on anaerobic fermentation of domestic refuse

Anaerobic fermentation of organic solid waste can provide a significant source of fuel gas (methane). Application of this process requires a better understanding of the kinetics of the biological system. The literature is replete with kinetic studies of this process as applied to waste solids from water pollution control systems. Much of this work has been conducted in the mesophilic temperature range. Increased temperatures yield higher reaction rates that will improve the economics of the process. The rate limiting step in the fermentation of refuse is the hydrolysis of the complex organic solids, in particular cellulose. Cellulose is a major component of the refuse. A laboratory study employing domestic refuse has shown the effect of temperature on the rate of methane fermentation. The optimum mesophilic temperature was found to be 42°C, while the optimum thermophilic temperature was at least 60°C. No data was obtained beyond the 60°C temperature. Reaction rate constants are presented for anaerobic fermentation of domestic refuse. Because of the characteristics of the substrate it^?was not possible to obtain the necessary measurements for evaluation of constants in the Monod model. An overall system constant was developed.     

Psychrophilic methane fermentation of excess sludge by enrichment culture

This study was carried out to determine the possibility of the psychrophilic methane fermentation of the excess sludge produced from fish-processing wastewater treatment. Three different kinds of seed sludge, named M seed sludge, K seed sludge and KL seed sludge were used for batch methane fermentation in Erlenmeyer flasks. M seed sludge was mesophilic, while the other two were prepared in our laboratory by soil collection to obtain a psychrophilic biological community. The effects of temperature on the total volume of biogas generated and on the rate of biogas generation were examined. The activity of K seen sludge with respect to gas yield at 15°C was found to be 1.6-fold higher than that of M seed sludge. Enrichment techniques were then used to accumulate psychrophilic bacteria in KL seed sludge, and the activity at 15°C increased by about two times due to these techniques. Comparison of the three kinds of seed sludge indicates that methane fermentation at low temperature may be feasible, and therefore further study to isolate the psychrophilic bacteria in anaerobic digestion is needed.     

‘Should I stay or should I go?’ Bacterial attachment vs biofilm formation on surface-modified membranes

A number of techniques are used for testing the anti-biofouling activity of surfaces, yet the correlation between different results is often questionable. In this report, the correlation between initial bacterial deposition (fast tests, reported previously) and biofilm growth (much slower tests) was analyzed on a pristine and a surface-modified reverse osmosis membrane ESPA-1. The membrane was modified with grafted hydrophilic polymers bearing negatively charged, positively charged and zwitter-ionic moieties. Using three different bacterial strains it was found that there was no general correlation between the initial bacterial deposition rates and biofilm growth on surfaces, the reasons being different for each modified surface. For the negatively charged surface the slowest deposition due to the charge repulsion was eventually succeeded by the largest biofilm growth, probably due to secretion of extracellular polymeric substances (EPS) that mediated a strong attachment. For the positively charged surface, short-term charge attraction by quaternary amine groups led to the fastest deposition, but could be eventually overridden by their antimicrobial activity, resulting in non-consistent results where in some cases a lower biofilm formation rate was observed. The results indicate that initial deposition rates have to be used and interpreted with great care, when used for assessing the anti-biofouling activity of surfaces. However, for a weakly interacting ‘low-fouling’ zwitter-ionic surface, the positive correlation between initial cell deposition and biofilm growth, especially under flow, suggests that for this type of coating initial deposition tests may be fairly indicative of anti-biofouling potential.     

Investigation of DNA orientation on gold by EC-STM.

The immobilization of thiol-derivatized DNA on a Au (111) single crystal surface by self-assembly has been investigated by electrochemical scanning tunneling microscopy (EC-STM). Continuous potential-dependent orientation changes of double-stranded oligodeoxynucleotides (ODN) have been observed in a certain potential range from 200 to 600 mV (versus SCE). It is suggested that the DNA duplexes stand straight on the gold surface at potentials negative of the potential of zero charge (pzc) and then lay down on the surface when the potential shifts positively. These results are in agreement with the expectation based on the Coulombic interaction consideration between negatively charged DNA helices and gold surface. As the applied potential shifts positively, the surface charge changes from negative to positive, that is, the Coulombic force between negatively charged DNA helices and gold surfaces changes from repulsion to attraction. However, for the single-stranded oligodeoxynucleotides, no distinct changes in the surface structure were observed with the applied potential.     

Biomethanation: Anaerobic fermentation of CO2, H2 and CO to methane

Studies to examine the microbial fermentation of coal gasification products (CO₂, H₂ and CO) to methane have been done with a mixed culture of anaerobic bacteria selected from an anaerobic sewage digestor. The specific rate of methane production at 37°C reached 25 mmol/g cell hr. The stoichiometry for methane production was 4 mmol H₂/mol CO₂. Cell recycle was used to increase the cell concentration from 2.5 to 8.3 g/liter; the volumetric rate of methane production ran from 1.3 to 4 liter/liter hr. The biogasification was also examined at elevated pressure (450 psi) and temperature to facilitate interfacing with a coal gasifier. At 60°C, the specific rate of methane production reached 50 mmol/g cell hr. Carbon monoxide utilization by the mixed culture of anaerobes and by a Rhodopseudomonas species was examined. Both cultures are able to carry out the shift conversion of CO and water to CO₂ and hydrogen.     

Evaluation of startup and operation of four anaerobic processes treating a synthetic meat waste

Two continuous stirred tanks reactors (CSTR) and four anaerobic fluidized bed reactors (AFBR) were used to study the treatment of a synthetic meat waste during single-and two-stage anaerobic treatment. Four configurations were investigated; a single-stage CSTR and AFBR and the two-stage systems CSTR-AFBR and AFBR-AFBR. Startup of the anaerobic reactors was achieved within 50 days by use of a regime that included stepped increases in influent COD, methanol substitution of the substrate, and addition of essential trace metals such as cobalt and nickel. Two-stage reactors removed up to 85% of influent COD concentrations of 5000 mg/L, whereas the single-stage AFBR and CSTR removed 76 and 9%, respectively. The proportion of methane in the effluent gases increased as the influent COD concentration was increased. Volumetric production of methane was greatest for the first stage of the AFBR-AFBR system. Solids retention times calculated for the AFBRs ranged from 7 to 12 days, sufficient to support methanogenesis. The AFBRs and two-stage systems were more resistant to an influent pH shock from the operating value of pH 6.8 down to pH 3 than the CSTRs and single-stage reactors. It was concluded that high-rate anaerobic treatment systems were applicable to meat industry wastewaters and that two-stage digestion produced a better quality effluent.     

Enhancement of biochemical methane potential from excess sludge with low organic content by mild thermal pretreatment

Excess sludge with low organic content always led to the failure of anaerobic digestion for methane production. Recently, the mild thermal pretreatment, which is usually operated at temperatures below 120 °C, has drawn much attention due to less energy consumption and no chemical addition. In this study the effect of mild thermal pretreatment (50–120 °C) on the solubilization and methane potential of excess sludge with a low concentration of organic matters was investigated. Experimental results showed that the concentration of soluble organic matters increased gradually with temperature during the mild thermal pretreatment of excess sludge. Biochemical methane potential experiments demonstrated that the potential of methane production from excess sludge was greatly enhanced by mild thermal pretreatment, and under the conditions of pretreatment temperature 100 °C and digestion time 20 d the methane yield was as high as 142.6 ± 2.5 mL/g of volatile solids. Mechanism investigation on the enhancement of methane production from excess sludge exhibited that the consumptions of sludge protein and carbohydrate, the adenosine 5′-triphosphate content of anaerobic microorganisms, the activities of key enzymes related to anaerobic digestion, and the amount of methanogens were all improved by mild thermal pretreatment, in correspondence with the production of methane.     

Interaction Between Equally Charged Membrane Surfaces Mediated by Positively and Negatively Charged Macro-Ions

In biological systems, charged membrane surfaces are surrounded by charged molecules such as electrolyte ions and proteins. Our recent experiments in the systems of giant phospholipid vesicles indicated that some of the blood plasma proteins (macro-ions) may promote adhesion between equally charged membrane surfaces. In this work, theory was put forward to describe an IgG antibody-mediated attractive interaction between negatively charged membrane surfaces which was observed in experiments on giant phospholipid vesicles with cardiolipin-containing membranes. The attractive interactions between negatively charged membrane surfaces in the presence of negatively and positively charged spherical macro-ions are explained using functional density theory and Monte Carlo simulations. Both, the rigorous solution of the variational problem within the functional density theory and the Monte Carlo simulations show that spatial and orientational ordering of macro-ions may give rise to an attractive interaction between negatively charged membrane surfaces. It is also shown that the distinctive spatial distribution of the charge within the macro-ions (proteins) is essential in this process.