Soybean molasses-based bioindicator system for monitoring sterilization process: Designing and performance evaluation

A novel cost-effective Bacillus atrophaeus Sterilization Bioindicator System (BIS) with a high quality and performance was developed from a soybean byproduct and compared with the commercial BIS. It was composed of recovery medium and dry-fermented spores with sand as the support. The BIS was developed and optimized using a sequential experimental design strategy. The recovery medium contained soluble starch (1.0 g/L), soybean molasses (30.0 g/L), tryptone (40.0 g/L), and bromothymol blue (0.02 g/L) at pH 8.5. The solid-state fermentation conditions of the bioreactor and environmental humidity had no significant effects on the spore yield and dry-heat resistance. The only substrate mineral that showed a positive effect was Mn²⁺, allowing Mg²⁺, K⁺, and Ca²⁺ to be eliminated from the formulation. Validation of optimized medium indicated D _160°C = 6.8±1.0 min (3.6 min more than the minimum) and spore yield = 2.3 ± 0.5 × 10⁹ CFU/g dry sand (10,000 × initial values). BIS performance resulted in D _160°C = 6.6 ± 0.1 min. Sporulation and germination kinetics allowed the sporulation process to be reduced to three days, and the growth of heat-damaged spores was sufficient to achieve visual identification of a non-sterile BIS within 21 h. Process economics was a minimum of 23.9%, and process cycle time was reduced from 29 to 15 days. The new BIS parameters demonstrated compliance to all regulatory requirements. No studies have yet described a BIS production from soybean molasses.     

Lipid productivity and fatty acid composition-guided selection of Chlorella strains isolated from Malaysia for biodiesel production

The need to develop biomass-based domestic production of high-energy liquid fuels (biodiesel) for transportation can potentially be addressed by exploring microalgae with high lipid content. Selecting the strains with adequate oil yield and quality is of fundamental importance for a cost-efficient biofuel feedstock production based on microalgae. This work evaluated 29 strains of Chlorella isolated from Malaysia as feedstock for biodiesel based on volumetric lipid productivity and fatty acid profiles. Phylogenetic studies based on 18S rRNA gene revealed that majority of the strains belong to true Chlorella followed by Parachlorella. The strains were similarly separated into two groups based on fatty acid composition. Of the 18 true Chlorella strains, Chlorella UMACC187 had the highest palmitic acid (C16:0) content (71.3?±?4.2 % total fatty acids, TFA) followed by UMACC84 (70.1?±?0.7 %TFA), UMACC283 (63.8?±?0.7 %TFA), and UMACC001 (60.3?±?4.0 %TFA). Lipid productivity of the strains at exponential phase ranged from 34.53 to 230.38 mg L^?1 day^?1, with Chlorella UMACC050 attaining the highest lipid productivity. This study demonstrated that Chlorella UMACC050 is a promising candidate for biodiesel feedstock production.     

From crude glycerol to carotenoids by using a Rhodotorula glutinis mutant

In this work eighteen red yeasts were screened for carotenoids production on glycerol containing medium. Strain C2.5t1 of Rhodotorula glutinis, that showed the highest productivity, was UV mutagenized. Mutant 400A15, that exhibited a 280 % increase in β–carotene production in respect to the parental strain, was selected. A central composite design was applied to 400A15 to optimize carotenoids and biomass productions. Regression analyses of the quadratic polynomial equations obtained (R² = 0.87 and 0.94, for carotenoids and biomass, respectively) suggest that the models are reliable and significant (P < 0.0001) in the prediction of carotenoids and biomass productions on the basis of the concentrations of crude glycerol, yeast extract and peptone. Accordingly, total carotenoids production achieved (14.07 ± 1.45 mg l^?1) under optimized growth conditions was not statistically different from the maximal predicted (14.64 ± 1.57 mg l^?1) (P < 0.05), and it was about 100 % higher than that obtained under un-optimized conditions. Therefore mutant 400A15 may represent a biocatalyst of choice for the bioconversion of crude glycerol into value-added metabolites, and a tool for the valorization of this by-product of the biodiesel industry.     

The membraneless bioelectrochemical reactor stimulates hydrogen fermentation by inhibiting methanogenic archaea

The membraneless bioelectrochemical reactor (Ml-BER) is useful for dark hydrogen fermentation. The effect of the electrochemical reaction on microorganisms in the Ml-BER was investigated using glucose as the substrate and compared with organisms in a membraneless non-bioelectrochemical reactor (Ml-NBER) and bioelectrochemical reactor (BER) with a proton exchange membrane. The potentials on the working electrode of the Ml-BER and BER with membrane were regulated to ?0.9 V (versus Ag/AgCl) to avoid water electrolysis with a carbon electrode. The Ml-BER showed suppressed methane production (19.8?±?9.1 mg-C·L^?1·day^?1) and increased hydrogen production (12.6?±?3.1 mg-H·L^?1·day^?1) at pH_out 6.2?±?0.1, and the major intermediate was butyrate (24.9?±?2.4 mM), suggesting efficient hydrogen fermentation. In contrast, the Ml-NBER showed high methane production (239.3?±?17.9 mg-C·L^?1·day^?1) and low hydrogen production (0.2?±?0.0 mg-H·L^?1·day^?1) at pH_out 6.3?±?0.1. In the cathodic chamber of the BER with membrane, methane production was high (276.3?±?20.4 mg-C·L^?1·day^?1) (pH_out, 7.2?±?0.1). In the anodic chamber of the BER with membrane (anode-BER), gas production was low because of high lactate production (43.6?±?1.7 mM) at pH_out 5.0?±?0.1. Methanogenic archaea were not detected in the Ml-BER and anode-BER. However, Methanosarcina sp. and Methanobacterium sp. were found in Ml-NBER. Prokaryotic copy numbers in the Ml-BER and Ml-NBER were similar, as were the bacterial community structures. Thus, the electrochemical reaction in the Ml-BER affected hydrogenotrophic and acetoclastic methanogens, but not the bacterial community.     

<Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> Spore Production Under Solid-State Fermentation of Lignocellulosic Residues

This study was conducted to elucidate cultivation conditions determining Bacillus amyloliquefaciens B-1895 growth and enhanced spore formation during the solid-state fermentation (SSF) of agro-industrial lignocellulosic biomasses. Among the tested growth substrates, corncobs provided the highest yield of spores (47?×?10¹⁰ spores g^?1 biomass) while the mushroom spent substrate and sunflower oil mill appeared to be poor growth substrates for spore formation. Maximum spore yield (82?×?10¹⁰ spores g^?1 biomass) was achieved when 15 g corncobs were moistened with 60 ml of the optimized nutrient medium containing 10 g peptone, 2 g KH₂PO₄, 1 g MgSO₄·7H₂O, and 1 g NaCl per 1 l of distilled water. The cheese whey usage for wetting of lignocellulosic substrate instead water promoted spore formation and increased the spore number to 105?×?10¹⁰ spores g^?1. Addition to the cheese whey of optimized medium components favored sporulation process. The feasibility of developed medium and strategy was shown in scaled up SSF of corncobs in polypropylene bags since yield of 10?×?10¹¹ spores per gram of dry biomass was achieved. In the SSF of lignocellulose, B. amyloliquefaciens B-1895 secreted comparatively high cellulase and xylanase activities to ensure good growth of the bacterial culture.     

Microalgal cultivation in porous substrate bioreactor for extracellular polysaccharide production

Netrium digitus is a representative of the species-rich class Zygnematophyceae (Streptophyta). Its intensive extracellular polysaccharide (EPS) production makes this alga interesting for biotechnological applications with a focus on cosmetics and food additives. Quantitative data on growth and EPS production in suspension and, for the first time, in immobilized culture using lab-scale porous substrate bioreactors, so-called Twin-Layer (TL) systems, is presented. It is shown that the cell as well as the EPS dry weight content is increased at least sixfold in immobilized compared to suspension culture. Due to the high amount of EPS, the biofilms reach a thickness of more than 8 mm after 27 days at 70 μmol photons m^?2 s^?1 and with 1.5% CO₂ supply. Frequent exchange of the growth medium results in a linear cell biomass increase of 2.02?±?0.09 g m^?2 growth area day^?1 compared to 2.99?±?0.09 g m^?2 day^?1, when the medium is not exchanged. Under this mode of cultivation, the EPS production is lower and a final concentration of 12.18?±?1.25 g m^?2 compared to 20.76?±?0.85 g m^?2, when medium was exchanged, is reached. It is clearly demonstrated that the relatively slow growing, but excessively EPS producing, microalgal species N. digitus can be grown in porous substrate bioreactors and that this culturing technique is a promising alternative to suspension culture for the Zygnematophyceae.     

Inorganic carbon and pH effect on growth and lipid productivity of Tetraselmis suecica and Chlorella sp (Chlorophyta) grown outdoors in bag photobioreactors

There has been considerable interest in cultivation of green microalgae (Chlorophyta) as a source of lipid that can alternatively be converted to biodiesel. However, almost all mass cultures of algae are carbon-limited. Therefore, to reach a high biomass and oil productivities, the ideal selected microalgae will most likely need a source of inorganic carbon. Here, growth and lipid productivities of Tetraselmis suecica CS-187 and Chlorella sp were tested under various ranges of pH and different sources of inorganic carbon (untreated flue gas from coal-fired power plant, pure industrial CO₂, pH-adjusted using HCl and sodium bicarbonate). Biomass and lipid productivities were highest at pH 7.5 (320?±?29.9 mg biomass L^?1 day^?1and 92?±?13.1 mg lipid L^?1 day^?1) and pH 7 (407?±?5.5 mg biomass L^?1 day^?1 and 99?±?17.2 mg lipid L^?1 day^?1) for T. suecica CS-187 and Chlorella sp, respectively. In general, biomass and lipid productivities were pH 7.5?>?pH 7?>?pH 8?>?pH 6.5 and pH 7?>?pH 7.5?=?pH 8?>?pH 6.5?>?pH 6?>?pH 5.5 for T. suecica CS-187 and Chlorella sp, respectively. The effect of various inorganic carbon on growth and productivities of T. suecica (regulated at pH?=?7.5) and Chlorella sp (regulated at pH?=?7) grown in bag photobioreactors was also examined outdoor at the International Power Hazelwood, Gippsland, Victoria, Australia. The highest biomass and lipid productivities of T. suecica (51.45?±?2.67 mg biomass L^?1 day^?1 and 14.8?±?2.46 mg lipid L^?1 day^?1) and Chlorella sp (60.00?±?2.4 mg biomass L^?1 day^?1 and 13.70?±?1.35 mg lipid L^?1 day^?1) were achieved when grown using CO₂ as inorganic carbon source. No significant differences were found between CO₂ and flue gas biomass and lipid productivities. While grown using CO₂ and flue gas, biomass productivities were 10, 13 and 18 %, and 7, 14 and 19 % higher than NaHCO₃, HCl and unregulated pH for T. suecica and Chlorella sp, respectively. Addition of inorganic carbon increased specific growth rate and lipid content but reduced biomass yield and cell weight of T. suecica. Addition of inorganic carbon increased yield but did not change specific growth rate, cell weight or content of the cell weight of Chlorella sp. Both strains showed significantly higher maximum quantum yield (F_v/F_m) when grown under optimum pH.     

Optimization of protease production by Streptomyces sp. A6 using statistical approach for reclamation of shellfish waste

Protease producing Streptomyces sp. A6 was isolated from intertidal zone of the coast of Diu (Gujarat, India). Plackett–Burman method was applied to identify important factors (shrimp waste, FeCl₃, ZnSO₄ and pH) influencing protease production by Streptomyces sp. A6. Further optimization was done by response surface methodology using central composite design. The concentrations of medium components for higher protease production as optimized using the above approach were (g l^?1): Shrimp waste, 14; FeCl₃, 0.035; ZnSO₄, 0.065 and pH, 8.0. This statistical optimization approach led to production of 129.02 ± 2.03 U ml^?1 of protease which was 4.96 fold higher compared to that obtained using the unoptimized medium. The protease production was scaled to 3 l in a 5-l bench fermenter using optimized medium which further increased the production by 63.4%. Deproteinization and chitin recovery obtained at the end of fermentation was 85.12 ± 4.7 and 70.58 ± 1.33%, respectively. The present study is the first report on statistical optimization of medium components for production of protease by Streptomyces species using cheaper raw material such as shrimp waste. The study also explored the possibility Streptomyces sp. A6 for reclamation of shrimp wastes.     

Effects of different nitrogen, phosphorus, and iron concentrations and salinity on lipid production in newly isolated strain of the tropical green microalga, Scenedesmus dimorphus KMITL

The fatty acid composition, the effect of different concentrations of nitrogen (16.5-344 mg ?L^?1), phosphorus (9–45 mg? L^?1), iron (9–45 mg? L^?1) and salinity levels (0–20 psu) on lipid production in the green microalga Scenedesmus dimorphus KMITL, a new strain isolated from a tropical country, Thailand, were studied. The alga was isolated from a freshwater fish pond, and cultured in Chlorella medium by varying one parameter at a time. The main fatty acid composition of this strain was C16–C18 (97.52 %) fatty acids. A high lipid content was observed in conditions of 16.5 mg? L^?1-N, or 22 mg ?L^?1-P, or 45 mg ?L^?1-Fe, or 5 psu salinity, which accumulated lipids to 20.3?±?0.4, 19.4?±?0.2, 24.7?±?0.5, and 14.3?±?0.2 % of algal biomass, respectively. Increasing lipid content and lipid productivity was noted when the alga was cultured under high iron concentration and high salinity, as well as under reduced phosphorus conditions, whereas nitrogen limitation only resulted in an increased lipid content.     

Enhancing ethanol production from cellulosic sugars using <Emphasis Type="Italic">Scheffersomyces (Pichia) stipitis</Emphasis>

Studies were performed on the effect of CaCO₃ and CaCl₂ supplementation to fermentation medium for ethanol production from xylose, glucose, or their mixtures using Scheffersomyces (Pichia) stipitis. Both of these chemicals were found to improve maximum ethanol concentration and ethanol productivity. Use of xylose alone resulted in the production of 20.68 ± 0.44 g L^?1 ethanol with a productivity of 0.17 ± 0.00 g L^?1 h^?1, while xylose plus 3 g L^?1 CaCO₃ resulted in the production of 24.68 ± 0.75 g L^?1 ethanol with a productivity of 0.21 ± 0.01 g L^?1 h^?1. Use of xylose plus glucose in combination with 3 g L^?1 CaCO₃ resulted in the production of 47.37 ± 0.55 g L^?1 ethanol (aerobic culture), thus resulting in an ethanol productivity of 0.39 ± 0.00 g L^?1 h^?1. These values are 229 % of that achieved in xylose medium. Supplementation of xylose and glucose medium with 0.40 g L^?1 CaCl₂ resulted in the production of 44.84 ± 0.28 g L^?1 ethanol with a productivity of 0.37 ± 0.02 g L^?1 h^?1. Use of glucose plus 3 g L^?1 CaCO₃ resulted in the production of 57.39 ± 1.41 g L^?1 ethanol under micro-aerophilic conditions. These results indicate that supplementation of cellulosic sugars in the fermentation medium with CaCO₃ and CaCl₂ would improve economics of ethanol production from agricultural residues.     

Effects of negative air ions on oxygen uptake kinetics,recovery and performance in exercise: a randomized,double-blinded study

Limited research has suggested that acute exposure to negatively charged ions may enhance cardio-respiratory function, aerobic metabolism and recovery following exercise. To test the physiological effects of negatively charged air ions, 14 trained males (age: 32?±?7 years; \( \overset{\cdotp }{V}{\mathrm{O}}_{2 \max } \) : 57?±?7 mL min^?1 kg^?1) were exposed for 20 min to either a high-concentration of air ions (ION: 220?±?30?×?10³ ions cm^?3) or normal room conditions (PLA: 0.1?±?0.06?×?10³ ions cm^?3) in an ionization chamber in a double-blinded, randomized order, prior to performing: (1) a bout of severe-intensity cycling exercise for determining the time constant of the phase II \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) response (τ) and the magnitude of the \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) slow component (SC); and (2) a 30-s Wingate test that was preceded by three 30-s Wingate tests to measure plasma [adrenaline] (ADR), [nor-adrenaline] (N-ADR) and blood [lactate] (B_Lac) over 20 min during recovery in the ionization chamber. There was no difference between ION and PLA for the phase II \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) τ (32?±?14 s vs. 32?±?14 s; P?=?0.7) or \( \overset{\cdotp }{V}{\mathrm{O}}_2 \) SC (404?±?214 mL vs 482?±?217 mL; P?=?0.17). No differences between ION and PLA were observed at any time-point for ADR, N-ADR and B_Lac as well as on peak and mean power output during the Wingate tests (all P?>?0.05). A high-concentration of negatively charged air ions had no effect on aerobic metabolism during severe-intensity exercise or on performance or the recovery of the adrenergic and metabolic responses after repeated-sprint exercise in trained athletes.     

Optimization of biomass and fatty acid productivity of Scenedesmus obliquus as a promising microalga for biodiesel production

Nowadays, microalgae are discussed as a promising feedstock for biodiesel production. The present study examines the possibility of enhancement of fatty acid productivity of Scenedesmus obliquus by modifications of the culture medium composition. The effect of different concentrations of sodium bicarbonate, salinity, potassium nitrate, glycerol and sugarcane molasses on the enhancement of biomass and esterified fatty acids production was studied. NaHCO₃ caused an increase in the biomass productivity at low concentrations (0.5 g L^?1), while negatively affected fatty acid productivity at all tested concentrations. Increase of salinity enhanced both biomass and fatty acid productivity. The optimum NaCl concentration and sea water ratio were 0.94 g L^?1 and 25 % which resulted in 56 and 39 % increase in fatty acid productivity, respectively. Nitrogen deficiency showed increase in fatty acid content by 54 % over control but fatty acid productivity was decreased as a result of growth inhibition. Nitrogen-free cultures and cultures treated with ?50 % concentrations of KNO₃ showed 96 and 42 % decrease in EFA productivity, respectively, as compared with the control. Addition of 0.05 and 0.1 M of glycerol increased the biomass productivity by 6 and 5 %, respectively but showed no significant effect on fatty acid productivity as a result of decrease in fatty acid content. Finally, usage of sugarcane molasses stimulated both biomass and fatty acid content. The increase in fatty acid productivity was 32, 65 and 73 % above the control level at 1, 3 and 5 g L^?1 of sugarcane molasses, respectively.     

High yield production of extracellular recombinant levansucrase by Bacillus megaterium

In this study, a high yield production bioprocess with recombinant Bacillus megaterium for the production of the extracellular enzyme levansucrase (SacB) was developed. For basic optimization of culture parameters and nutrients, a recombinant B. megaterium reporter strain that produced green fluorescent protein under control of a vector-based xylose-inducible promoter was used. It enabled efficient microtiter plate-based screening via fluorescence analysis. A pH value of pH?6, 20 % of dissolved oxygen, 37 °C, and elevated levels of biotin (100 μg?L^?1) were found optimal with regard to high protein yield and reduced overflow metabolism. Among the different compounds tested, fructose and glycerol were identified as the preferred source of carbon. Subsequently, the settings were transferred to a B. megaterium strain recombinantly producing levansucrase SacB based on the plasmid-located xylose-inducible expression system. In shake flask culture under the optimized conditions, the novel strain already secreted the target enzyme in high amounts (14 U?mL^?1 on fructose and 17.2 U?mL^?1 on glycerol). This was further increased in high cell density fed-batch processes up to 55 U?mL^?1, reflecting a levansucrase concentration of 0.52 g?L^?1. This is 100-fold more than previous efforts for this enzyme in B. megaterium and more than 10-fold higher than reported values of other extracellular protein produced in this microorganism so far. The recombinant strain could also handle raw glycerol from biodiesel industry which provided the same amount and quality of the recombinant protein and suggests future implementation into existing biorefinery concepts.     

Continuous enzymatic biodiesel production from coconut oil in two-stage packed-bed reactor incorporating an extracting column to remove glycerol formed as by-product

The transesterification of coconut oil with ethanol catalyzed by Burkholderia cepacia lipase immobilized on polysiloxane–polyvinyl alcohol was performed in a continuous flow. The experimental design consisted of a two-stage packed-bed reactor incorporating a column with cationic resin (Lewatit GF 202) to remove the glycerol formed as by-product and the reactor performance was quantified for three different flow rates corresponding to space-times from 10 to 14 h. The influence of space-time on the ethyl ester (FAEE) concentrations, yields and productivities was determined. The reactor operation was demonstrated for space-time of 14 h attaining FAEE concentrations of 58.5 ± 0.87 wt%, FAEE yields of 97.3 ± 1.9 % and productivities of 41.6  ± 1.0 mg_ester g _medium ^?1  h^?1. Biodiesel purified samples showed average kinematic viscosity values of 5.5 ± 0.3 mm² s^?1 that meet the criteria established by the American National Standard ASTM (D6751). The immobilized lipase was found to be stable regarding its morphological and catalytic characteristics, showing half-life time (t _1/2) around 1540 h. The continuous packed-bed reactor connected in series with simultaneous glycerol removal has a great potential to attain high level of transesterification yields, raising biodiesel productivity.     

The citric acid production from raw glycerol by Yarrowia lipolytica yeast and its regulation

The optimal cultivation conditions ensuring the maximal rate of citric acid (CA) biosynthesis by glycerol-grown mutant Yarrowia lipolytica NG40/UV7 were found to be as follows: growth limitation by inorganic nutrients (nitrogen, phosphorus, or sulfur), 28 °C, pH 5.0, dissolved oxygen concentration (pO₂) of 50 % (of air saturation), and pulsed addition of glycerol from 20 to 80 g L^?1 depending on the rate of medium titration. Under optimal conditions of fed-batch cultivation, in the medium with pure glycerol, strain Y. lipolytica NG40/UV7 produced 115 g L^?1 of CA with the mass yield coefficient of 0.64 g g^?1 and isocitric acid (ICA) amounted to 4.6 g L^?1; in the medium with raw glycerol, CA production was 112 g L^?1 with the mass yield coefficient of 0.90 g g^?1 and ICA amounted to 5.3 g L^?1. Based on the activities of enzymes involved in the initial stages of raw glycerol assimilation, the tricarboxylic acid cycle and the glyoxylate cycle, the mechanism of increased CA yield from glycerol-containing substrates in Y. lipolytica yeast was explained.     

Factors affecting biohydrogen production by unicellular halotolerant cyanobacterium Aphanothece halophytica

The effects of several physiological parameters on H₂ production rate in the unicellular halotolerant cyanobacterium Aphanothece halophytica were investigated. Under nitrogen deprivation, the growth of cells was inhibited, but H₂ production rate was enhanced approximately fourfold. Interestingly, cells grown under sulfur deprivation exhibited a decrease in cell growth, H₂ production rate, and bidirectional hydrogenase activity. Glucose was the preferred sugar source for H₂ production by A. halophytica, but H₂ production decreased at high glucose concentrations. H₂ production rate was optimum when cells were grown in the presence of 0.75 M?NaCl, or 0.4 μM?Fe³⁺, or 1 μM?Ni²⁺. The optimum light intensity and temperature for H₂ production were 30 μmol photons m^?2?s^?1 and 35 °C, respectively. A two-stage culture of A. halophytica was performed in order to overcome the reduction of cell growth in N-free medium. In the first stage, cells were grown in normal medium to accumulate biomass, and in the second stage, H₂ production by the obtained biomass was induced by growing cells in N-free medium supplemented with various chemicals for 24 h. A. halophytica grown in N-free medium containing various MgSO₄ concentrations had a high H₂ production rate between 11.432 and 12.767 μmol H₂ mg?chlorophyll a (chl a)^?1?h^?1, a 30-fold increase compared to cells grown in normal medium. The highest rate of 13.804 μmol H₂ mg?chl a ^?1?h^?1 was obtained when the N-free growth medium contained 0.4 μM Fe³⁺. These results suggested the possibility of using A. halophytica and some other halotolerant cyanobacteria thriving under extreme environmental conditions in the sea as potential sources for H₂ production in the future.     

Successful operation of continuous reactors at short retention times results in high-density,fast-rate Dehalococcoides dechlorinating cultures

The discovery of Dehalococcoides mccartyi reducing perchloroethene and trichloroethene (TCE) to ethene was a key landmark for bioremediation applications at contaminated sites. D. mccartyi-containing cultures are typically grown in batch-fed reactors. On the other hand, continuous cultivation of these microorganisms has been described only at long hydraulic retention times (HRTs). We report the cultivation of a representative D. mccartyi-containing culture in continuous stirred-tank reactors (CSTRs) at a short, 3-d HRT, using TCE as the electron acceptor. We successfully operated 3-d HRT CSTRs for up to 120 days and observed sustained dechlorination of TCE at influent concentrations of 1 and 2 mM TCE to ≥97 % ethene, coupled to the production of 10¹² D. mccartyi cells L_culture ^?1. These outcomes were possible in part by using a medium with low bicarbonate concentrations (5 mM) to minimize the excessive proliferation of microorganisms that use bicarbonate as an electron acceptor and compete with D. mccartyi for H₂. The maximum conversion rates for the CSTR-produced culture were 0.13?±?0.016, 0.06?±?0.018, and 0.02?±?0.007 mmol Cl^? L_culture ^?1 h^?1, respectively, for TCE, cis-dichloroethene, and vinyl chloride. The CSTR operation described here provides the fastest laboratory cultivation rate of high-cell density Dehalococcoides cultures reported in the literature to date. This cultivation method provides a fundamental scientific platform for potential future operations of such a system at larger scales.     

Decolorization of azo dyes by Geobacter metallireducens

Geobacter metallireducens was found to be capable of decolorizing several azo dyes with different structures to various extents. Pyruvate, ethanol, acetate, propionate, and benzoate could support 66.3?±?2.6?93.7?±?2.1 % decolorization of 0.1 mM acid red 27 (AR27) in 40 h. The dependence of the specific decolorization rate on AR27 concentration (25 to 800 μM) followed Michaelis–Menten kinetics (K _m?=?186.9?±?1.4 μΜ, V _max?=?0.65?±?0.02 μmol?mg protein^?1 h^?1). Enhanced AR27 decolorization was observed with the increase of cell concentrations ranging from 7.5 to 45 mgL^?1. AR27 decolorization by G. metallireducens was retarded by the presence of goethite, which competed electrons with AR27 and was reduced to Fe(II). The addition of low concentrations of humic acid (1?100 mgL^?1) or 2-hydroxy–1,4-naphthoquinone (0.5?50 μM) could improve the decolorization performance of G. metallireducens. High-performance liquid chromatography analysis suggested reductive pathway to be responsible for decolorization. This was the first study on azo dye decolorization by Geobacter strain and might improve our understanding of natural attenuation and bioremediation of environments polluted by azo dyes.     

Improvement of Biomethane Production from Sewage Sludge in Co-digestion with Glycerol and Waste Frying Oil,Using a Design of Experiments

A central composite design circumscribed method was used to define the experimental conditions that improve the methane production rate (k_CH4, liters of methane per kilogram of VS of waste added and per day) and the cumulative methane production (cMP, liters of methane per kilogram of VS of waste added) of the co-digestion of sewage sludge (SS) with crude glycerol (cGly) and waste frying oil (WFO). Three factors were selected, i.e., SS concentration, global co-substrate concentration, and mass fraction of cGly (x_cGly) in a mixture of cGly and WFO (in chemical oxygen demand, COD). SS digestion without co-substrate reached a cMP of (294?±?6) L·kg^?1 and a k_CH4 of (64?±?1) L·kg^?1·d^?1, at standard temperature and pressure conditions and expressed relatively to the initial volatile solids. After statistical analysis, SS and co-substrate concentrations of 4.6 g·L^?1 and 8.8 g·L^?1 (in COD), respectively, with x_cGly of 0.8, were defined to simultaneously boost cMP (91 % more) and k_CH4 (3-fold increase). Application of these conditions would yield 214 MWh more in electricity per 1000 m³ of SS digested.     

Ulva lactuca and U. flexuosa (Chlorophyta,Ulvophyceae) cultivation in Brazilian tropical waters: recruitment,growth, and ulvan yield

Ulva spp. are used in a wide range of commercial applications, including bioremediation, food, bioenergy, pharmaceuticals, and agriculture. The sulfated polysaccharide ulvan obtained from Ulva spp. is of interest for triggering plant defenses against disease. However, the cultivation of Ulva spp. is still in its infancy. This study verified the feasibility of cultivating Ulva lactuca and Ulva flexuosa at two sites on the tropical Brazilian coast. We investigated the following: (a) methods to induce sporulation, (b) comparison of seeding ropes inoculated in vitro versus seeding at sea over 40 days, (c) production and harvest cycles at 15 and 30 days, (d) growth productivity of U. flexuosa at sea and in outdoor tanks, and (e) comparison of ulvan yields from biomass cultivated in tanks and the sea. High nutrient treatment was the most efficient method to induce sporulation (7,540?±?3,133 spores m^?1). Sea-based cultivation of U. flexuosa was only successful at one site. Seeding of ropes in vitro was more efficient than seeding at sea (0.31?±?0.20 g m^?2 day^?1), and 15-day harvest cycles were most efficient (20.1?±?1.8 % day^?1; 0.46?±?0.11 g m^?2 day^?1). Despite differences in plant growth in tanks (27.9?±?4.4 % day^?1) and at sea (20.1?±?1.8 % day^?1), the dry biomass and ulvan yields (17.7?±?5.0 %) did not differ between these systems. Cultivation of U. flexuosa was feasible at sea using in vitro seeding with a production cycle of 15 days in Brazilian tropical waters and tanks with high irradiance and enriched seawater.