Batch and continuous fermentative production of hydrogen with anaerobic sludge entrapped in a composite polymeric matrix

Cell immobilization techniques were adopted to biohydrogen production using immobilized anaerobic sludge as the seed culture. Sucrose-based synthetic wastewater was converted to H₂ using batch and continuous cultures. A novel composite polymeric material comprising polymethyl methacrylate (PMMA), collagen, and activated carbon was used to entrap biomass for H₂ production. Using the PMMA immobilized cells, the favorable conditions for batch H₂ fermentation were 35 °C, pH 6.0, and an 20 g COD l⁻¹ of sucrose, giving a H₂ production rate of 238 ml h⁻¹ l⁻¹ and a H₂ yield of 2.25 mol H₂ mol sucrose⁻¹. Under these optimal conditions, continuous H₂ fermentation was conducted at a hydraulic retention time (HRT) of 4–8 h, giving the best H₂-producing rate of 1.8 l h⁻¹ l⁻¹ (over seven-fold of the best batch result) at a HRT of 6 h and a H₂ yield of 2.0 mol H₂ mol sucrose⁻¹. The sucrose conversion was essentially over 90% in all runs. The biogas consisted of only H₂ and CO₂. The major soluble metabolites were butyric acid, acetic acid, and 2,3-butandiol, while a small amount of ethanol also detected. The PMMA-immobilized-cell system developed in this work seems to be a promising H₂-producing process due to the high stability in continuous operations and the capability of achieving a competitively high H₂ production rate under a relatively low organic loading rate.     

Sulfate reduction from phosphogypsum using a mixed culture of sulfate-reducing bacteria

Phosphogypsum (CaSO₄), a primary by-product of phosphoric acid production, is accumulated in large stockpiles and occupies vast areas of land. It poses a severe threat to the quality of water and land in countries producing phosphoric acid. In this study, the potential of sulfate-reducing bacteria for biodegradation of this sulfur-rich industrial solid waste was assessed. The effect of phosphogypsum concentration, carbon and nitrogen sources, temperature, pH and stirring on the growth of sulfate-reducing bacteria was investigated. Growth of sulfate-reducing bacteria was monitored by measuring sulfide production. Phosphogypsum was shown to be a good source of sulfate, albeit that the addition of organic carbon was necessary for bacterial growth. Biogenic sulfide production occurred with phosphogypsum up to a concentration of 40 g L⁻¹, above which no growth of sulfate-reducing bacteria was observed. Optimal growth was obtained at 10 g L⁻¹ phosphogypsum. Both the gas mixture H₂/CO₂ and lactate supported high amounts of H₂S formation (19 and 11 mM, respectively). The best source of nitrogen for sulfate-reducing bacteria was yeast extract, followed by ammonium chloride. The presence of nitrate had an inhibitory effect on the process of sulfate reduction. Stirring the culture at 150 rpm slightly stimulated H₂S formation, probably by improving sulfate solubility.     

The first crenarchaeon capable of growth by anaerobic carbon monoxide oxidation coupled with H2 production

The ability to grow by anaerobic CO oxidation with production of H₂ from water is known for some thermophilic bacteria, most of which belong to Firmicutes, as well as for a few hyperthermophilic Euryarchaeota isolated from deep-sea hydrothermal habitats. A hyperthermophilic, neutrophilic, anaerobic filamentous archaeon strain 1505 = VKM B-3180 = KCTC 15798 was isolated from a terrestrial hot spring in Kamchatka (Russia) in the presence of 30% CO in the gas phase. Strain 1505 could grow lithotrophically using carbon monoxide as the energy source with the production of hydrogen according to the equation CO + H₂O → CO₂ + H₂; mixotrophically on CO plus glucose; and organotrophically on peptone, yeast extract, glucose, sucrose, or Avicel. The genome of strain 1505 was sequenced and assembled into a single chromosome. Based on 16S rRNA gene sequence analysis and in silico genome-genome hybridization, this organism was shown to be closely related to the Thermofilum adornatum species. In the genome of Thermofilum sp. strain 1505, a gene cluster (TCARB_0867-TCARB_0879) was found that included genes of anaerobic (Ni,Fe-containing) carbon monoxide dehydrogenase and genes of energy-converting hydrogenase ([Ni,Fe]-CODH–ECH gene cluster). Compared to the [Ni,Fe]-CODH–ECH gene clusters occurring in the sequenced genomes of other H₂-producing carboxydotrophs, the [Ni,Fe]-CODH–ECH gene cluster of Thermofilum sp. strain 1505 presented a novel type of gene organization. The results of the study provided the first evidence of anaerobic CO oxidation coupled with H₂ production performed by a crenarchaeon, as well as the first documented case of lithotrophic growth of a Thermofilaceae representative.     

Functional consortium for hydrogen production from cellobiose: Concentration-to-extinction approach

A functional bacterial consortium that can effectively hydrolyze cellobiose and produce bio-hydrogen was isolated by a concentration-to-extinction approach. The sludge from a cattle feedlot manure composting plant was incubated with 2.5–20 g l^?1 cellobiose at 35 °C and pH 6.0. The microbial diversity of serially concentrated suspensions significantly decreased following increasing cellobiose concentration, finally leaving only two viable strains, Clostridium butyricum strain W4 and Enterococcus saccharolyticus strain. This consortium has a maximum specific hydrogen production rate of 2.19 mol H₂ mol hexose^?1 at 5 g l^?1 cellobiose. The metabolic pathways shifted from ethanol-type to acetate-butyrate type as cellobiose concentration increased from 2.5 to >7 g l^?1. The concentration-to-extinction approach is effective for isolating functional consortium from natural microflora. In this case the functional strains of interest are more tolerant to the increased loadings of substrates than the non-functional strains.     

Evaluation of the association of mercury(II) with some dicysteinyl tripeptides

The present study was undertaken to gain insight into the associations of mercury(II) with dicysteinyl tripeptides in buffered media at pH 7.4. We investigated the effects of increasing the distance between cysteinyl residues on mercury(II) associations and complex formations. The peptide–mercury(II) formation constants and their associated thermodynamic parameters in 3-(N-morpholino)propanesulfonic acid (MOPS) buffered solutions were evaluated by isothermal titration calorimetry. Complexes formed in different relative ratios of mercury(II) to cysteinyl peptides in ammonium formate buffered solutions were characterized by LTQ Orbitrap mass spectrometry. The results from these studies show that n-alkyl dicysteinyl peptides (CP 1–4), and an aryl dicysteinyl peptide (CP 5) can serve as effective “double anchors” to accommodate the coordination sites of mercury(II) to form predominantly one-to-one Hg(peptide) complexes. The aryl dicysteinyl peptide (CP 5) also forms the two-to-two Hg₂(peptide)₂ complex. In the presence of excess peptide, Hg(peptide)₂ complexes are also detected. Notably, increasing the distance between the ligating groups or “anchor points” in CP 1–5 does not significantly affect their affinity for mercury(II). However, the enthalpy change (ΔH) values (ΔH₁ ∼ −91 kJ mol⁻¹ and ΔH₂ ∼ −66 kJ mol⁻¹) for complex formation between CP 4 and 5 with mercury(II) are about one and a half times larger than the related values for CP 1, 2 and 3 (ΔH₁ ∼ −66 kJ mol⁻¹ and ΔH₂ ∼ 46 kJ mol⁻¹). The corresponding entropy change (ΔS) values (ΔS₁ ∼ −129 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −116 J K⁻¹ mol⁻¹) of the structurally larger dicysteinyl peptides CP 4 and 5 are less entropically favorable than for CP 1, 2 and 3 (ΔS₁ ∼ −48 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −44 J K⁻¹ mol⁻¹). Generally, these associations result in a decrease in entropy, indicating that these peptide–mercury complexes potentially form highly ordered structures. The results from this study show that dicysteinyl tripeptides are effective in binding mercury(II) and they are promising motifs for the design of multi-cysteinyl peptides for binding more than one mercury(II) ion per peptide.     

The sensitivity of water chemistry to climate in a forested,nitrogen-saturated catchment recovering from acidification

Fluxes of major ions and nutrients were measured in the N-saturated mountain forest catchment-lake system of Čertovo Lake (Czech Republic) from 1998 to 2014. The lake has been rapidly recovering from atmospheric acidification due to a 90% decrease in sulphate (SO₄²⁻) deposition since the late 1980s and nitrate (NO₃⁻) contribution to the pool of strong acid anion and leaching of dissolved organic carbon (DOC) have increased. Present concentrations of base cations, phosphorus (P), total organic N (TON), and ionic (Al_i) and organically bound (Al_o) aluminium in tributaries are thus predominantly governed by NO₃⁻ and DOC leaching. Despite a continuing recovery lasting 25 years, the Čertovo catchment is still a net source of protons (H⁺), producing 44 mmol m⁻² yr⁻¹ H⁺ on a catchment-area basis (corresponding to 35 μmol L⁻¹ on a concentration basis). Retention of the deposited inorganic N in the catchment averages 20%, and ammonium consumption (51 mmol m⁻² yr⁻¹) and net NO₃⁻ production (28 mol m⁻² yr⁻¹) are together the dominant terrestrial H⁺ generating processes. In contrast, the importance of SO₄²⁻ release from the soils on terrestrial H⁺ production is continuously decreasing, with an average of 47 mmol m⁻² yr⁻¹ during the study. The in-lake biogeochemical processes reduce the incoming acidity by ∼40%, neutralizing 23 μmol L⁻¹ H⁺ (i.e., 225 mmol m⁻² yr⁻¹ on a lake-area basis). Denitrification and photochemical and microbial decomposition of DOC are the most important in-lake H⁺ consuming processes (50 and 39%, respectively), while hydrolysis of Al_i (from tributaries and photochemically liberated from Al_o) is the dominant in-lake H⁺ generating process. Because the trends in water chemistry and H⁺ balance in the catchment-lake system are increasingly related to variability in NO₃⁻ and DOC leaching, they have become sensitive to climate-related factors (drought, elevated runoff) and forest damage that significantly modify the leaching of these anions. During the study period, increased exports of NO₃⁻ (accompanied by Al_i and base cations) from the Čertovo catchment occurred after a dry and hot summer, after forest damage, and during elevated winter runoff. Increasing DOC export due to decreasing acid deposition was further elevated during years with higher runoff (and especially during events with lateral flow), and was accompanied by P, TON, and Al_o leaching. The climate-related processes, which originally “only” confounded chemical trends in waters recovering from acidification, may soon become the dominant variables controlling water composition in N-saturated catchments.     

Antioxidant activity and protection of human umbilical vein endothelial cells from hydrogen peroxide-induced injury by DMC,a chalcone from buds of Cleistocalyx operculatus

The aim of this work was to study the antioxidant activity and the protective effect of 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone (DMC), the main compound from the buds of Cleistocalyx operculatus, on human umbilical vein endothelial cells against cytotoxicity induced by H₂O₂. The antioxidant activities of DMC were measured by ABTS assay, ferric reducing antioxidant power (FRAP) and hydroxyl radical scavenging activity, and protective effects of DMC on human umbilical vein endothelial cells against cytotoxicity induced by H₂O₂ were tested. DMC was found to have high ABTS radical scavenging activity (176.5 ± 5.2 μmol trolox equivalents/500 μmol DMC) and strong ferric reducing antioxidant power (213.3 ± 5.8 μmol trolox equivalents/500 μmol DMC). In addition, DMC scavenged the hydroxyl radicals, with IC₅₀ values of 243.7 ± 6.3 μM, slightly lower than the reference antioxidant ascorbic acid (ASC). Moreover, DMC could protect the human umbilical vein endothelial cells against H₂O₂-induced cytotoxicity by decrease intracellular and extracellular ROS levels, reduction in catalase (CAT) activity and increment in malondialdehyde (MDA) level. These results suggested that DMC has the potential to be used in the therapy of oxidative damage.     

Comparison of batch,fed-batch and continuous well-mixed reactors for enzymatic hydrolysis of orange peel wastes

An alternative potential feedstock for bioethanol in the automotive sector is citrus peel waste (CPW), which can be processed through enzymatic hydrolysis and fermentation. The present work considers mathematical modeling of orange peel wastes (OPW) hydrolysis with the use of free enzymes and compares the performance of batch, fed-batch and continuous well-mixed reactors after introducing appropriate rate equations in dynamic mass balances. MATLAB^® was used for model implementation.Following the Michaelis–Menten approach, the authors used their own kinetic parameters for the pectin hydrolysis rate equation. The parameters were generated in an apposite experimental program for OPW hydrolysis to galacturonic acid with consideration of product inhibition; the corresponding values were obtained after Lineweaver–Burk linearization and are: r_max = 0.28 g/(L min), K_m = 19.80 g/L and K_IGA = 6.96 g/L, respectively. Vice-versa, the authors adopted the Kadam's group kinetic schemes and parameters for cellulose hydrolysis to cellobiose and glucose. The mathematical model of a well-mixed batch reactor was perfectly validated against the experimental results of OPW hydrolysis to galacturonic acid. In the case of a continuous well-mixed reactor, high dilution rates determine low conversion of OPW. The increased complication of fed-batch operation does not add advantages when compared to batch processing.     

Purification,kinetic and thermodynamic characterization of soluble acid invertase from sugarcane (Saccharum officinarum L.)

We report for the first time kinetic and thermodynamic properties of soluble acid invertase (SAI) of sugarcane (Saccharum officinarum L.) salt sensitive local cultivar CP 77-400 (CP-77). The SAI was purified to apparent homogeneity on FPLC system. The crude enzyme was about 13 fold purified and recovery of SAI was 35%. The invertase was monomeric in nature and its native molecular mass on gel filtration and subunit mass on SDS-PAGE was 28 kDa. SAI was highly acidic having an optimum pH lower than 2. The acidic limb was missing. Proton transfer (donation and receiving) during catalysis was controlled by the basic limb having a pKa of 2.4. Carboxyl groups were involved in proton transfer during catalysis. The kinetic constants for sucrose hydrolysis by SAI were determined to be: k_m = 55 mg ml^?1, k_cat = 21 s^?1, k_cat/k_m = 0.38, while the thermodynamic parameters were: ΔH* = 52.6 kJ mol^?1, ΔG* = 71.2 kJ mol^?1, ΔS* = ?57 J mol^?1 K^?1, ΔG*_E–S = 10.8 kJ mol^?1 and ΔG*_E–T = 2.6 kJ mol^?1. The kinetics and thermodynamics of irreversible thermal denaturation at various temperatures 53–63 °C were also determined. The half -life of SAI at 53 and 63 °C was 112 and 10 min, respectively. At 55 °C, surprisingly the half -life increased to twice that at 53 °C. ΔG*, ΔH* and ΔS* of irreversible thermal stability of SAI at 55 °C were 107.7 kJ mol^?1, 276.04 kJ mol^?1 and 513 J mol^?1K^?1, respectively.     

Effects of flooding on photosynthesis and root respiration in saltcedar (Tamarix ramosissima), an invasive riparian shrub

The introduced shrub Tamarix ramosissima invades riparian zones, but loses competitiveness under flooding. Metabolic effects of flooding could be important for T. ramosissima, but have not been previously investigated. Photosynthesis rates, stomatal conductance, internal (intercellular) CO₂, transpiration, and root alcohol dehydrogenase (ADH) activity were compared in T. ramosissima across soil types and under drained and flooded conditions in a greenhouse. Photosynthesis at 1500 μmol quanta m⁻² s⁻¹ (A₁₅₀₀) in flooded plants ranged from 2.3 to 6.2 μmol CO₂ m⁻² s⁻¹ during the first week, but A₁₅₀₀ increased to 6.4–12.7 μmol CO₂ m⁻² s⁻¹ by the third week of flooding. Stomatal conductance (g_s) at 1500 μmol quanta m⁻² s⁻¹ also decreased initially during flooding, where g_s was 0.018 to 0.099 mol H₂O m⁻² s⁻¹ during the first week, but g_s increased to 0.113–0.248 mol H₂O m⁻² s⁻¹ by the third week of flooding. However, photosynthesis in flooded plants was reduced by non-stomatal limitations, and subsequent increases indicate metabolic acclimation to flooding. Root ADH activities were higher in flooded plants compared to drained plants, indicating oxygen stress. Lower photosynthesis and greater oxygen stress could account for the susceptibility of T. ramosissima at the onset of flooding. Soil type had no effect on photosynthesis or on root ADH activity. In the field, stomatal conductance, leaf water potential, transpiration, and leaf δ¹³C were compared between T. ramosissima and other flooded species. T. ramosissima had lower stomatal conductance and water potential compared to Populus deltoides and Phragmites australis. Differences in physiological responses for T. ramosissima could become important for ecological concerns.     

Seasonal variations in photosynthetic irradiance response curves of macrophytes from a Mediterranean coastal lagoon

The main photosynthesis and respiration parameters (dark respiration rate, light saturated production rate, saturation irradiance, photosynthetic efficiency) were measured on a total of 23 macrophytes of the Thau lagoon (2 Phanerogams, 5 Chlorophyceae, 10 Rhodophyceae and 6 Phaeophyceae). Those measurements were performed in vitro under controlled conditions, close to the natural ones, and at several seasons. Concomitantly, measurements of pigment concentrations, carbon, phosphorous and nitrogen contents in tissues were performed. Seasonal intra-specific variability of photosynthetic parameters was found very high, enlightening an important acclimatation capacity. The highest photosynthetic capacities were found for Chlorophyceae (e.g. Monostroma obscurum thalli at 17 °C, 982 μmol O₂ g⁻¹ dw h⁻¹ and 9.1 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹, respectively for light saturated net production rate and photosynthetic efficiency) and Phanerogams (e.g. Nanozostera noltii leaves at 25 °C, 583 μmol O₂ g⁻¹ dw h⁻¹ and 2.6 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹ respectively for light saturated net production rate and photosynthetic efficiency). As expected, species with a high surface/volume ratio were found to be more productive than coarsely branched thalli and thick blades shaped species. Contrary to R_d (ranging 6.7–794 μmol O₂ g⁻¹ dw h⁻¹, respectively for Rytiphlaea tinctoria at 7 °C and for Dasya sessilis at 25 °C) for which a positive relationship with water temperature was found whatever the species studied, the evolution of P/I curves with temperature exhibited different responses amongst the species. The results allowed to show summer nitrogen limitation for some species (Gracilaria bursa-pastoris and Ulva spp.) and to propose temperature preferences based on the photosynthetic parameters for some others (N. noltii, Zostera marina, Chaetomorpha linum).     

Biohydrogen production from palm oil mill effluent using immobilized Clostridium butyricum EB6 in polyethylene glycol

A novel polyethylene glycol (PEG) gel was fabricated and used as a carrier to immobilize Clostridium butyricum EB6 to improve biohydrogen (bio-H₂) production from palm oil mill effluent (POME). POME is used as a substrate that can act as a carbon source. The resulting PEG-immobilized cells were found to yield 5.35 LH₂/L-POME, and the maximum H₂ production rate was 510 mL H₂/L-POME h (22.7 mmol/L h). The Monod-type kinetic model was used to describe the effect of substrate (POME) concentration on the H₂ production rate. The acclimation of immobilized cells greatly improved H₂ production. Batch experiments demonstrated that particle size of PEG-immobilized cells for efficient H₂ production 3 mm. It is significant that this is the first report on whole-cell immobilization in PEG for H₂ production from POME.     

Co-digestion of cow manure,food waste and intermittent input of fat

Pulses of oil were added to completely mixed reactors fed with dairy cow manure and food waste, after achieving a stable performance at an organic loading rate of 4.6 ± 0.1 gCOD/(l_reactor day), an oily waste effluent from a canned fish processing industry was fed in the form of pulses. The oil concentration rose up to 9, 12, 15 and 18 gCOD_oil/l_reactor, after the pulse feeding in the reactor. The highest fat concentration of 18 gCOD_oil/l_reactor promoted a persistent inhibition in the process of the continuous reactor, although in batch assays, the reactor content evidenced a capacity to degrade more oil and to degrade the accumulated organic matter. All the other pulses had a positive effect in the methane production. From a practical point of view, this work demonstrates that controlled intermittent inputs of oil can enhance the methane production in a co-digestion of cow manure and food waste.     

Synthesis,crystal structure and magnetic properties of the spin crossover system [Fe(pq)3]2+

Three new compounds formulated (ClO₄)₂[Fe(pq)₃] (1), (BF₄)₂[Fe(pq)₃] · EtOH (2) and {(ClO₄)[MnCr(C₂O₄)₃][Fe(pq)₂(H₂O)₂]} (3), where pq is 2,2′-pyridylquinoline, have been synthesised and characterised. Despite the different crystal packing exhibited by 1 and 2, the cationic species [Fe(pq)₃]²⁺ are structurally quite similar. At 293 K, the Fe–N bond lengths are characteristic of the iron(II) in the high-spin state. In contrast to 1, 2 undergoes a continuous spin transition. Indeed, at 95 K its structure experiences a noticeable change in the Fe–N bonds and angles, i.e. the Fe–N bonds shorten by 0.194 Å on the average. The magnetic behaviour confirms that 1 is fully high-spin in the 4–300 K temperature range while 2 shows a spin transition centred at T_1/2 = 150 K. The corresponding enthalpy, entropy and interaction parameter are ΔH = 7.49 kJ mol^?1, ΔS = 50 J K^?1 mol^?1and Γ = 1.35 kJ mol^?1. Compound 3 has been obtained as a microcrystalline powder. The magnetic properties of 3 point at the occurrence of ferromagnetic coupling below 100 K and the onset of a ferromagnetic ordering below 10 K (Weiss constant equal to 6.8 K). The Mössbauer spectra of 3 show the occurrence of a magnetic order at T ? 4.2 K.     

Continuous biomethanization of agrifood industry waste: A case study in Spain

The anaerobic digestion technology is a biological treatment widely used to reduce the pollution load of wet waste biomass. In this work we present the results obtained by performing extensive experiments of anaerobic digestion of slaughterhouse waste, tomato industry waste and olive oil industry waste in continuous mode, which were designed to demonstrate that anaerobic digestion is an effective technology from an environmental and economic point of view.Biogas yields obtained are between 35.22 and 5.45 Nm³ biogas/m³ olive oil industry waste and tomato industry waste respectively and the slaughterhouse wastes achieve intermediate production, 30.86 Nm³ biogas/m³ municipal slaughterhouse waste and 22.53 Nm³ biogas/m³ Iberian pig slaughterhouse waste. Moreover, it possible to degrade between 63.46 and 75.3% of the initial organic matter.If these results are analyzed, the environmental, energetic economic benefits of anaerobic digestion can be quantified. Biomethanation of all these wastes generated annually in Extremadura could prevent the emission of 134,772 t of equivalent carbon dioxide, generate an energy similar to that provided by 2826 toe and reach payback times from 3.29 to 3.75 years for anaerobic digestion plant designed to treat the wastes generated by a medium-sized industry. So, we have fulfilled all the planned aims.     

Effect of rare earth elements on in vitro rumen microbial fermentation and feed digestion

The objectives of this study were to investigate the effects of rare earth elements (REEs) on in vitro rumen fermentation, gas production, microbial protein synthesis and nutrient digestion using in vitro batch culture and continuous culture technique. A mixture of REE containing (g/kg) 380 g of LaCI₃·6H₂O, 521 g of CeCI₃·6H₂O, 30 g of PrCI₃·6H₂O and 69 g chlorides of other light REEs. The experimental diet consisted of 885 g/kg barley grain, 84 g/kg barley silage and 31 g/kg supplement (dry matter (DM) basis). Diet supplemented with different dosages of REE (control, no additional REE; low, 400 mg/kg REE; and high, 800 mg/kg REE, DM basis) were incubated for 4, 8, 14 and 24 h in diluted rumen fluid. At the end of 24 h of incubation, gas production and concentration of volatile fatty acid (VFA) linearly increased with increasing REE supplementation; whereas, influence of REE supplementation on VFA profile was marginal. Dry matter disappearance was not affected (P>0.10). Six dual-flow continuous culture fermenters were used in a replicated 3 × 3 Latin square with same treatments and same diet used in the batch culture. Mean ruminal pH (5.71) and total VFA (93.6 mM) concentration were not affected by supplementation of REE. The molar proportion (mol/100 mol) of acetate (39.1) and propionate (50.5) was similar among the treatments. However, the proportion (mol/100 mol) of butyrate was higher with the high REE (6.6) than with low REE (5.3) or the control (5.8). Ruminal true digestibilities of organic matter (OM) (0.785, 0.811 and 0.828), acid detergent fibre (0.360, 0.431 and 0.432) and crude protein (0.496, 0.590 and 0.589) for control, low and high REE, respectively, linearly increased with increasing REE supplementation, whereas, the increase in ruminal digestibility from low to high dosage of REE was minimal. Microbial nitrogen (N) production (g/day) and microbial efficiency (g N/kg of truly fermented OM) were not affected by treatments. Improvement of ruminal digestibility of OM due to REE supplementation was attributed to the increase in digestibility of fibre and degradability of protein. The results suggest that REE supplementation improved ruminal fibrolytic and proteolytic activities.     

Enhanced methane production from wool textile residues by thermal and enzymatic pretreatment

Methane production from two types of wool textile wastes (TW1 and TW2) was investigated. To improve the digestibility of these textiles, different pretreatments were applied, and comprised thermal treatment (at 120 °C for 10 min), enzymatic hydrolysis (using an alkaline endopeptidase at different levels of enzymatic loading, at 55 °C for 0, 2, and 8 h), and a combination of these two treatments. Soluble protein concentration and sCOD (soluble chemical oxygen demand) were measured to evaluate the effectivity of the different pretreatment conditions to degrade wool keratin. The sCOD as well as the soluble protein content had increased in both textile samples in comparison to untreated samples, as a response to the different pretreatments indicating breakdown of the wool keratin structure.The combined treatments and the thermal treatments were further evaluated by anaerobic batch digestion assays at 55 °C. Combined thermal and enzymatic treatment of TW1 and TW2 resulted in methane productions of 0.43 N m³/kg VS and 0.27 N m³/kg VS, i.e., 20 and 10 times higher yields, respectively, than that gained from untreated samples. The application of thermal treatment by itself was less effective and resulted in increasing the methane production by 10-fold for TW1 and showing no significant improvement for TW2.     

Comparative diel oxygen cycles preceding and during a Karenia bloom in Sarasota Bay,Florida, USA

The diel change in dissolved oxygen concentrations were recorded with an automated incubator containing a pulsed oxygen sensor in Sarasota Bay, Florida. The deployments occurred during a ‘pre-bloom’ period in May to June 2006, and during a harmful algal bloom dominated by Karenia brevis in September 2006. The diurnal (daylight) increase in dissolved oxygen concentrations varied from 16 to 104 μmol O₂ l⁻¹ with the corresponding nocturnal decrease in oxygen varying from 16 to 77 μmol O₂ l⁻¹. Nocturnal respiration consumed 42–113% of the diurnal net oxygen production with the minimum and maximum during the pre-bloom period. Hourly production rates closely followed fluctuations in irradiance with maximum rates in the late morning. Hourly oxygen utilization rates (community respiration) at night were highest during the first few hours after sunset.     

Cholinium carboxylate ionic liquids for pretreatment of lignocellulosic materials to enhance subsequent enzymatic saccharification

The present study is the first report demonstrating that ionic liquids consisting of cholinium cations and linear carboxylate anions ([Ch][CA] ILs) can be used for pretreatment of lignocellulosic materials to enhance subsequent enzymatic saccharification. Six variants of [Ch][CA] ILs were systematically prepared by combining cholinium cations with linear monocarboxylate anions ([C_nH_2n+1–COO]⁻, n = 0–2) or dicarboxylate anions ([HOOC–C_nH_2n+1–COO]⁻, n = 0–2). These [Ch][CA] ILs were analyzed for their toxicity to yeast cell growth and their ability to pretreat kenaf powder for subsequent enzymatic saccharification. When assayed against yeast growth, the EC₅₀ for choline acetate ([Ch][OAc]) was 510 mM, almost one order of magnitude higher than that for 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]). The cellulose saccharification ratio after pretreatment at 110 °C for 16 h with [Ch][OAc] (100.6%) was almost comparable with that after pretreatment with [Emim][OAc]. Therefore, [Ch][OAc] is a biocompatible alternative to [Emim][OAc] for lignocellulosic material pretreatment.     

The mechanism(s) involved in the photoprotection of PSII at elevated CO2 in nodulated alfalfa plants

In a previous study, we found that enhanced CO₂ subjected to nodulated alfalfa plants grown at different temperatures (ambient and ambient + 4 °C) and water availability regimes could protect PSII from photodamage. The main objective of this study was to determine the mechanism(s) involved in the photoprotection of PSII at elevated CO₂ levels in this plant. Elevated CO₂ reduced carboxylation capacity-induced photosynthetic acclimation and reduced enzymatic and/or nonenzymatic antioxidant activities, suggesting that changes in electron flow did not cause any photooxidative damage (which was also confirmed by H₂O₂ and lipid peroxidation analyses). Enhanced nonphotochemical quenching and xanthophyll cycle pigments revealed that plants grown at 700 μmol mol⁻¹ CO₂ compensated for the reduction in energy sink with a larger capacity for nonphotochemical dissipation of excitation energy as heat, i.e., modulating the status of the VAZ components. Elevated CO₂ induced the de-epoxidation of violaxanthin to zeaxanthin, facilitating thermal dissipation and protecting the photosynthetic apparatus against the deleterious effect of excess excitation energy.