Phenol removal from hypersaline wastewaters in a Membrane Biological Reactor (MBR): operation and microbiological characterisation

In this study, two Membrane Biological Reactors (MBR) with submerged flat membranes, one at lab-scale conditions and the other at pilot-plant conditions, were operated at environmental temperature to treat an industrial wastewater characterised by low phenol concentrations (8-16 mg L⁻¹) and high salinity (∼150-160 mS cm⁻¹). During the operation of both reactors, the phenol loading rate was progressively increased and less than 1 mg phenol L⁻¹ was detected even at very low HRTs (0.5-0.7 days). Membrane fouling was minimized by the cross flow aeration rate inside the MBRs and by intermittent permeation. Microbial community analysis of both reactors revealed that members of the genera Halomonas and Marinobacter (gammaproteobacteria) were major components. Growth-linked phenol degradation by pure cultures of Marinobacter isolates demonstrated that this bacterium played a major role in the removal of phenol from the bioreactors.     

Hydrodynamic behaviors in fermentative hydrogen bioreactors by pressure fluctuation analysis

Three bioreactor configurations were employed in these investigations, which consisted of working volumes of 10, 1.2 and 1.2 L. Power spectrum diagrams of bed pressure fluctuation were used with hydraulic retention times (HRT) and geometric factors to identify the flow regimes in the bioreactors, where HRT varied from 8 to 1 h. It was found that the flow regimes in the bioreactors changed from a dispersed regime to coalesced and slugging regimes, when the biogas production rate (BPR) increased, as a result of decreasing the operating HRT. The flow regime was a dispersed bubble regime when the HRT was higher than 4 h in the bioreactor, whereas when the HRT was 2 h the coalesced bubble phenomena occurred in the bioreactor. A slugging regime was found when the HRT was lower than 1 h in thinner bioreactor.     

Biological treatment of refinery spent caustics under halo-alkaline conditions

The present research demonstrates the biological treatment of refinery sulfidic spent caustics in a continuously fed system under halo-alkaline conditions (i.e. pH 9.5; Na⁺ = 0.8 M). Experiments were performed in identical gas-lift bioreactors operated under aerobic conditions (80-90% saturation) at 35 °C. Sulfide loading rates up to 27 mmol L⁻¹ day⁻¹ were successfully applied at a HRT of 3.5 days. Sulfide was completely converted into sulfate by the haloalkaliphilic sulfide-oxidizing bacteria belonging to the genus Thioalkalivibrio. Influent benzene concentrations ranged from 100 to 600 μM. At steady state, benzene was removed by 93% due to high stripping efficiencies and biodegradation. Microbial community analysis revealed the presence of haloalkaliphilic heterotrophic bacteria belonging to the genera Marinobacter, Halomonas and Idiomarina which might have been involved in the observed benzene removal. The work shows the potential of halo-alkaliphilic bacteria in mitigating environmental problems caused by alkaline waste.     

Detecting novel SNPs and breed-specific haplotypes at calpastatin gene in Iranian fat- and thin-tailed sheep breeds and their effects on protein structure

Calpastatin has been introduced as a potential candidate gene for growth and meat quality traits. In this study, genetic variability was investigated in the exon 6 and its intron boundaries of ovine CAST gene by PCR-SSCP analysis and DNA sequencing. Also a protein sequence and structural analysis were performed to predict the possible impact of amino acid substitutions on physicochemical properties and structure of the CAST protein. A total of 487 animals belonging to four ancient Iranian sheep breeds with different fat metabolisms, Lori-Bakhtiari and Chall (fat-tailed), Zel-Atabay cross-bred (medium fat-tailed) and Zel (thin-tailed), were analyzed. Eight unique SSCP patterns, representing eight different sequences or haplotypes, CAST-1, CAST-2 and CAST-6 to CAST-11, were identified. Haplotypes CAST-1 and CAST-2 were most common with frequency of 0.365 and 0.295. The novel haplotype CAST-8 had considerable frequency in Iranian sheep breeds (0.129). All the consensus sequences showed 98–99%, 94–98%, 92–93% and 82–83% similarity to the published ovine, caprine, bovine and porcine CAST locus sequences, respectively. Sequence analysis revealed four SNPs in intron 5 (C24T, G62A, G65T and T69-) and three SNPs in exon 6 (c.197A > T, c.282G > T and c.296C > G). All three SNPs in exon 6 were missense mutations which would result in p.Gln 66 Leu, p.Glu 94 Asp and p.Pro 99 Arg substitutions, respectively, in CAST protein. All three amino acid substitutions affected the physicochemical properties of ovine CAST protein including hydrophobicity, amphiphilicity and net charge and subsequently might influence its structure and effect on the activity of Ca2 + channels; hence, they might regulate calpain activity and afterwards meat tenderness and growth rate. The Lori-Bakhtiari population showed the highest heterozygosity in the ovine CAST locus (0.802). Frequency difference of haplotypes CAST-10 and CAST-8 between Lori-Bakhtiari (fat-tailed) and Zel (thin-tailed) breeds was highly significant (P < 0.001), indicating that these two haplotypes might be breed-specific haplotypes that distinguish between fat-tailed and thin-tailed sheep breeds.     

Effects of biodrying process on municipal solid waste properties

In this paper, the effect of biodrying process on municipal solid waste (MSW) properties was studied. The results obtained indicated that after 14 d, biodrying reduced the water content of waste, allowing the production of biodried waste with a net heating value (NHV) of 16,779 ± 2,074 kJ kg⁻¹ wet weight, i.e. 41% higher than that of untreated waste. The low moisture content of the biodried material reduced, also, the potential impacts of the waste, i.e. potential self-ignition and potential odors production. Low waste impacts suggest to landfill the biodried material obtaining energy via biogas production by waste re-moistening, i.e. bioreactor. Nevertheless, results of this work indicate that biodrying process because of the partial degradation of the organic fraction contained in the waste (losses of 290 g kg⁻¹ VS), reduced of about 28% the total producible biogas.     

Effect of feed to inoculum ratios on biogas yields of food and green wastes

Biogas and methane yields of food and green wastes and their mixture were determined using batch anaerobic digesters at mesophilic (35 ± 2 °C) and thermophilic (50 ± 2 °C) temperatures. The mixture was composed of 50% food waste and 50% green waste, based on the volatile solids (VS) initially added to the reactors. The thermophilic digestion tests were performed with four different feed to inoculum (F/I) ratios (i.e., 1.6, 3.1, 4.0 and 5.0) and the mesophilic digestion was conducted at one F/I (3.1). The results showed that the F/I significantly affected the biogas production rate. At four F/Is tested, after 25 days of thermophilic digestion, the biogas yield was determined to be 778, 742, 784 and 396 mL/g VS for food waste, respectively; 631, 529, 524 and 407 mL/g VS for green waste, respectively; and 716, 613, 671 and 555 mL/g VS for the mixture, respectively. About 80% of the biogas production was obtained during the first 10 days of digestion. At the F/I of 3.1, the biogas and methane yields from mesophilic digestion of food waste, green waste and their mixture were lower than the yields obtained at thermophilic temperature. The biogas yields were 430, 372 and 358 mL/g VS, respectively, and the methane yields were 245, 206, and 185 mL/g VS, respectively.     

Activated zeolite—suitable carriers for microorganisms in anaerobic digestion processes?

Plant cell wall structures represent a barrier in the biodegradation process to produce biogas for combustion and energy production. Consequently, approaches concerning a more efficient de-polymerisation of cellulose and hemicellulose to monomeric sugars are required. Here, we show that natural activated zeolites (i.e. trace metal activated zeolites) represent eminently suitable mineral microhabitats and potential carriers for immobilisation of microorganisms responsible for anaerobic hydrolysis of biopolymers stabilising related bacterial and methanogenic communities. A strategy for comprehensive analysis of immobilised anaerobic populations was developed that includes the visualisation of biofilm formation via scanning electron microscopy and confocal laser scanning microscopy, community and fingerprint analysis as well as enzyme activity and identification analyses. Using SDS polyacrylamide gel electrophoresis, hydrolytical active protein bands were traced by congo red staining. Liquid chromatography/mass spectroscopy revealed cellulolytical endo- and exoglucanase (exocellobiohydrolase) as well as hemicellulolytical xylanase/mannase after proteolytic digestion. Relations to hydrolytic/fermentative zeolite colonisers were obtained by using single-strand conformation polymorphism analysis (SSCP) based on amplification of bacterial and archaeal 16S rRNA fragments. Thereby, dominant colonisers were affiliated to the genera Clostridium, Pseudomonas and Methanoculleus. The specific immobilisation on natural zeolites with functional microbes already colonising naturally during the fermentation offers a strategy to systematically supply the biogas formation process responsive to population dynamics and process requirements.     

Treatment of poultry slaughterhouse wastewater in hybrid upflow anaerobic sludge blanket reactor packed with pleated poly vinyl chloride rings

In this study, the performance of 5.4 L hybrid upflow anaerobic sludge blanket (HUASB) reactor for treating poultry slaughterhouse wastewater under mesophilic conditions (29-35 °C), was investigated. After starting-up, the reactor was loaded up to an OLR of 19 kg COD/m³ d and achieved varied TCOD and SCOD removal efficiencies of 70-86% and 80-92%, respectively. The biogas was varied between 1.1 and 5.2 m³/m³ d with the maximum methane content of 72%. The maximum methane yield was 0.32 m³/kg COD_removed at an OLR of 9.27 kg COD/m³ d. Black matured granules of size between 2.5 and 5 mm were observed at the end of 225 d operation. RTD study showed the flow behavior was in mixed regime at the end of performance study. Step wise polynomial regression analysis was fitted well. Methanobacterium and Methanosaeta bacteria were dominant at the end of start-up whereas Methanosarcina, Cocci and rods were predominant at the end of performance studies.     

Examination of microbial fuel cell start-up times with domestic wastewater and additional amendments

Rapid startup of microbial fuel cells (MFCs) and other bioreactors is desirable when treating wastewaters. The startup time with unamended wastewater (118 h) was similar to that obtained by adding acetate or fumarate (110-115 h), and less than that with glucose (181 h) or Fe(III) (353 h). Initial current production took longer when phosphate buffer was added, with startup times increasing with concentration from 149 h (25 mM) to 251 h (50 mM) and 526 h (100 mM). Microbial communities that developed in the reactors contained Betaproteobacteria, Acetoanaerobium noterae, and Chlorobium sp. Anode biomass densities ranged from 200 to 600 μg/cm² for all amendments except Fe(Ш) (1650 μg/cm²). Wastewater produced 91 mW/m², with the other MFCs producing 50 mW/m² (fumarate) to 103 mW/m² (Fe(III)) when amendments were removed. These experiments show that wastewater alone is sufficient to acclimate the reactor without the need for additional chemical amendments.     

Dry-thermophilic anaerobic digestion of simulated organic fraction of Municipal Solid Waste: Process modeling

Solid retention time (SRT) is a very important operational variable in continuous and semicontinuous waste treatment processes since the organic matter removal efficiency - expressed in terms of percentage of Dissolved Organic Carbon (% DOC) or Volatile Solids (% VS) removed - and the biogas or methane production are closely related with the SRT imposed. Optimum SRT is depending on the waste characteristics and the microorganisms involved in the process and, hence, it should be determined specifically in each case.In this work a series of experiments were carried out to determine the effect of SRT, from 40 to 8 days, on the performance of the dry (30% Total Solids) thermophilic (55 °C) anaerobic digestion of organic fraction of Municipal Solid Wastes (OFMSW) operating at semicontinuous regime of feeding.The experimental results show than 15 days is the optimum SRT (the best between all proved) for this process. Besides, data of organic matter concentration and methane production versus SRT have been used to obtain the kinetic parameters of the kinetic model of Romero García (1991): the maximum specific growth rate of the microorganisms (μ_max = 0.580 days⁻¹) and the fraction of substrate non-biodegradable (α = 0.268).     

Emergy analysis of biogas production and electricity generation from small-scale agricultural digesters

An emergy analysis was performed to assess the relative sustainability and environmental impact of small-scale energy production using Taiwanese model plug-flow anaerobic digesters to treat livestock manure in Costa Rica. Emergy analysis quantifies all inputs to a system by converting them to solar energy equivalents, thus allowing for direct comparison of the diverse inputs of renewable energies, human labor and economic goods needed to construct and maintain anaerobic digestion systems. The digesters were located on the campus of EARTH University, Costa Rica, and the biogas was utilized to power a 40 kW generator that supplies electricity for farm operations. Separate emergy analyses were performed for the biogas production and the combination of biogas production and generation of electricity. Manure was the largest input in both analyses, accounting for 85.3% of the annual emergy input for biogas production and 66.9% for electricity generation from the biogas. The fraction of emergy inputs from renewable sources (ΦR) was 66% for biogas production and 52% for electricity generation from the biogas. The transformities of biogas and electricity generation from the biogas were 5.23E+04 sej/J and 1.01E+06 sej/J respectively. The emergy yield ratios (EYR) were 2.93 for biogas production and 2.07 for electricity generation indicating that these digesters efficiently match purchased resources and renewable energies to produce energy from livestock manure. The generation of electricity from the biogas resulted in a decrease in the emergy sustainability index (ESI) from 5.67 to 2.22 and an increase in the environmental loading ratio (ELR) from 0.52 to 0.93. Using a generator to convert the biogas to electricity does decrease the sustainability of the system, largely due to the high emergy value associated with the electrical generation equipment and machinery, but these results demonstrate that the production of biogas and the generation of electricity from Taiwanese model digesters in Costa Rica are environmentally sustainable processes that result in the production of energy that is largely dependent on renewable and recycled energies.     

Improved bio-energy yields via sequential ethanol fermentation and biogas digestion of steam exploded oat straw

Using standard laboratory equipment, thermochemically pretreated oat straw was enzymatically saccharified and fermented to ethanol, and after removal of ethanol the remaining material was subjected to biogas digestion. A detailed mass balance calculation shows that, for steam explosion pretreatment, this combined ethanol fermentation and biogas digestion converts 85-87% of the higher heating value (HHV) of holocellulose (cellulose and hemicellulose) in the oat straw into biofuel energy. The energy (HHV) yield of the produced ethanol and methane was 9.5-9.8 MJ/(kg dry oat straw), which is 28-34% higher than direct biogas digestion that yielded 7.3-7.4 MJ/(kg dry oat straw). The rate of biogas formation from the fermentation residues was also higher than from the corresponding pretreated but unfermented oat straw, indicating that the biogas digestion could be terminated after only 24 days. This suggests that the ethanol process acts as an additional pretreatment for the biogas process.     

Biogas production and saccharification of Salix pretreated at different steam explosion conditions

Different steam explosion conditions were applied to Salix chips and the effect of this pretreatment was evaluated by running both enzymatic hydrolysis and biogas tests. Total enzymatic release of glucose and xylose increased with pretreatment harshness, with maximum values being obtained after pretreatment for 10 min at 210 °C. Harsher pretreatment conditions did not increase glucose release, led to degradation of xylose and to formation of furfurals. Samples pretreated at 220 and 230 °C initially showed low production of biogas, probably because of inhibitors produced during the pretreatment, but the microbial community was able to adapt and showed high final biogas production. Interestingly, final biogas yields correlated well with sugar yields after enzymatic hydrolysis, suggesting that at least in some cases a 24 h enzymatic assay may be developed as a quick method to predict the effects of pretreatment of lignocellulosic biomass on biogas yields.     

Enhanced denitrification and organics removal in hybrid wetland columns: comparative experiments

This study investigated three lab-scale hybrid wetland systems with traditional (gravel) and alternative substrates (wood mulch and zeolite) for removing organic, inorganic pollutants and coliforms from a synthetic wastewater, in order to investigate the efficiency of alternative substrates, and monitor the stability of system performance. The hybrid systems were operated under controlled variations of hydraulic load (q, 0.3-0.9 m³/m² d), influent ammoniacal nitrogen (NH₄-N, 22.0-80.0 mg/L), total nitrogen (TN, 24.0-84.0 mg/L) and biodegradable organics concentration (BOD₅, 14.5-102.0 mg/L). Overall, mulch and zeolite showed promising prospect as wetland substrates, as both media enhanced the removal of nitrogen and organics. Average NH₄-N, TN and BOD₅ removal percentages were over 99%, 72% and 97%, respectively, across all three systems, indicating stable removal performances regardless of variable operating conditions. Higher Escherichia coli removal efficiencies (99.9%) were observed across the three systems, probably due to dominancy of aerobic conditions in vertical wetland columns of the hybrid systems.     

Optimization of biogas production from cattle manure by pre-treatment with ultrasound and co-digestion with crude glycerin

Biogas production by co-digestion of cattle manure with crude glycerin obtained from biodiesel production was studied after pre-treatment of the cattle manure or mixtures of cattle manure with different amounts of added glycerin with ultrasound. Batch experiments with 1750 mL of medium containing 1760 g of screened cattle manure or mixtures of cattle manure (screened or ground) and 70-140 mL or crude glycerin were incubated under mesophilic and thermophilic condition in stirred tank reactors. Under mesophilic conditions, the addition of 4% glycerin to screened manure increased biogas production by up to 400%. Application of sonication (20 kHz, 0.1 kW, and 4 min) to a mixture of manure + 4% glycerin increased production of biogas by up to 800% compared to untreated manure. The best results were obtained under thermophilic conditions using sonicated mixtures of ground cattle manure with 6% added glycerin (348 L methane/kg COD removed were obtained).     

Continuous hydrogen and butyric acid fermentation by immobilized Clostridium tyrobutyricum ATCC 25755: Effects of the glucose concentration and hydraulic retention time

The effects of the hydraulic retention time (HRT = 8, 10, 12 or 16.7 h) and glucose concentration (30, 40 or 50 g/L) on the production of hydrogen and butyrate by an immobilized Clostridium tyrobutyricum culture, grown under continuous culturing conditions, were evaluated. With 30 g/L glucose, the higher HRTs tested led to greater butyrate concentrations in the culture, i.e., 9.3 g/L versus 12.9 g/L with HRTs of 8 h and 16.7 h, respectively. In contrast, higher biogas and hydrogen production rates were generally seen when the HRT was lower. Experiments with different glucose concentrations saw a significant amount of glucose washed out when 50 g/L was used, the highest being 22.7 g/L when the HRT was 16.7 h. This study found the best conditions for the continuous production of hydrogen and butyric acid by C. tyrobutyricum to be with an HRT of 12 h and a glucose concentration of 50 g/L, respectively.     

Regeneration,colonisation and growth rates of allofragments in four common stream plants

Colonisation by stream plants occurs to a large extent from simple stem fragments. Allofragments are stem fragments formed by mechanical breakage. We studied regeneration, colonisation, and growth rates in four common stream plants: Elodea canadensis Michx., Myriophyllum spicatum L., Potamogeton perfoliatus L. and Ranunculus baudotii x pseudofluitans. The objectives of this study were to determine (1) if shoots with an apical tip have higher regeneration (growth of new shoots and rhizomes from allofragments) and colonisation (root attachment in sediment) abilities and higher relative growth rates (RGR) than shoots without an apical tip, and (2) if fragment size correlates with regeneration and colonisation abilities and with RGR of fragments. For all species, over 60% of fragments regenerated new shoots and colonised. Apical shoots and larger fragments generally had higher regeneration and colonisation abilities and higher RGR. Relative growth rate for E. canadensis and M. spicatum was between 0.06 and 0.09 d⁻¹ whereas it was about half this rate for Ranunculus and P. perfoliatus (0.02–0.04 d⁻¹).     

The effect of acid pretreatment on the anaerobic digestion and dewatering of waste activated sludge

Waste activated sludge (WAS) is difficult to degrade in anaerobic digestion systems and pretreatments have been shown to speed up the hydrolysis stage. Here the effects of acid pretreatment (pH 6-1) using HCl on subsequent digestion and dewatering of WAS have been investigated. Optimisation of acid dosing was performed considering digestibility benefits and level of acid required. Pretreatment to pH 2 was concluded to be the most effective. In batch digestion this yielded the same biogas after 13 days as compared to untreated WAS at 21 days digestion. In semi-continuous digestion experiments (12 day hydraulic retention time at 35 °C) it resulted in a 14.3% increase in methane yield compared to untreated WAS, also Salmonella was eradicated in the digestate. Dewatering investigations suggested that the acid pretreated WAS required 40% less cationic polymer addition to achieve the same cake solid content. A cost analysis was also carried out.     

Eutrophication-induced changes in benthic algae affect the behaviour and fitness of the marine amphipod Gammarus locusta

This study, conducted in mesocosms, natural field sites, and in laboratory aquaria, showed that eutrophication altered the nutrient status and dominance patterns among marine macroalgae, which in turn, stimulated gammaridean density. Gammaridean abundance correlated positively with both nutrient addition and the amount of green algae (also stimulated by nutrient enrichment). Path analysis indicated that the direct effect of nutrients on gammaridean density was of less importance than the indirect effect through increased production of green algae. In cage colonisation experiments, either in the field or in a control mesocosm kept under ambient nutrient conditions, more gammarids colonised nutrient enriched algae (E-algae) than algae with ambient nutrient levels (A-algae). Gammarus locusta generally grew faster on nutrient enriched algal specimens and when reared on green rather than on brown algae (fucoids). The nutrient status of periphytic algae did not affect gammaridean growth significantly, but the number of egg-carrying females (and thus egg production) was significantly higher among gammarids reared on E-periphyton. The gammaridean habitat preference order (red > green > brown > periphyton) was almost the reverse of their growth rate in feeding assays (periphyton > green > brown). This implies that macroalgae may be more important as a habitat than as a food source for these animals, which then have to become mobile in search of optimal food items. In this process, algal nutrient content was important as the gammarids in our study actively chose high quality nutrient-rich food, which, in addition, increased their fitness. Stimulated growth rates and egg production may ultimately lead to population increase, which, combined with the preference for high nutrient food items may dampen the initial effect of nutrient enrichment (i.e. blooms of green macroalgae) in shallow coastal waters.