Fernando G. Fermoso

Metal supplementation to UASB bioreactors: from cell-metal interactions to full-scale application

Upflow anaerobic sludge bed (UASB) bioreactors are commonly used for anaerobic wastewater treatment. Trace metals need to be dosed to these bioreactors to maintain microbial metabolism and growth. The dosing needs to balance the supply of a minimum amount of micronutrients to support a desired microbial activity or growth rate with a maximum level of micronutrient supply above which the trace metals become inhibitory or toxic. In studies on granular sludge reactors, the required micronutrients are undefined and different metal formulations with differences in composition, concentration and species are used. Moreover, an appropriate quantification of the required nutrient dosing and suitable ranges during the entire operational period has been given little attention. This review summarizes the state-of-the-art knowledge of the interactions between trace metals and cells growing in anaerobic granules, which is the main type of biomass retention in anaerobic wastewater treatment reactors. The impact of trace metal limitation as well as overdosing (toxicity) on the biomass is overviewed and the consequences for reactor performance are detailed. Special attention is given to the influence of metal speciation in the liquid and solid phase on bioavailability. The currently used methods for trace metal dosing into wastewater treatment reactors are overviewed and ways of optimization are suggested.

Biochemical methane potential of two-phase olive mill solid waste: Influence of thermal pretreatment on the process kinetics

The effect of thermal pretreatment on two-phase olive mill solid waste was evaluated by chemical oxygen demand solubilisation and biochemical methane potential (BMP) tests. Temperatures of 100, 120, 160 and 180°C were applied during 60, 120 and 180 min for each temperature studied. The highest chemical oxygen demand solubilisation after pretreatment (42%) was found for 120 and 180°C during 180 min in both cases. These two conditions were selected for the BMP tests. BMP tests showed two different stages: a first exponential stage and a sigmoidal zone after a lag period. No influence of the pretreatment was observed on the kinetic constant of the first-stage. Clear difference was observed in the maximum methane production rate of the second stage, 76.8 mL CH4/(g VS day) was achieved after pretreatment at 180°C (180 min), value 22% and 40% higher than that obtained for the untreated and pretreated OMSW at 120°C, respectively.

Dosing of anaerobic granular sludge bioreactors with cobalt: impact of cobalt retention on methanogenic activity.

The effect of dosing a metal limited anaerobic sludge blanket (UASB) reactor with a metal pulse on the methanogenic activity of granular sludge has thus far not been successfully modeled. The prediction of this effect is crucial in order to optimize the strategy for metal dosage and to prevent unnecessary losses of resources. This paper describes the relation between the initial immobilization of cobalt in anaerobic granular sludge cobalt dosage into the reactor and the evolution of methanogenic activity during the subsequent weeks. An operationally defined parameter (A0.B0) was found to combine the amount of cobalt immobilized instantaneously upon the pulse (B0) and the amount of cobalt immobilized within the subsequent 24h (A0). In contrast with the individual parameters A0 and B0, the parameter A0.B0 correlated significantly with the methanogenic activity of the sludge during the subsequent 16 or 35 days. This correlation between metal retention and activity evolution is a useful tool to implement trace metal dosing strategies for biofilm-based biotechnological processes.

Assessment of two-phase olive mill solid waste and microalgae co-digestion to improve methane production and process kinetics

Olive mill solid waste (OMSW) is a pollutant waste coming from olive oil elaboration by the two-phase centrifugation system. OMSW has a high organic matter content and unbalanced carbon to nitrogen (C/N) ratio, 31/1, which avoids obtaining high methane yields in the anaerobic digestion of this waste. In the present study a microalgae, Dunaliella salina, was employed as co-substrate for the OMSW anaerobic digestion in order to decrease the C/N ratio and increase its biodegradability. Different co-digestion mixtures (C/N ratios) were studied. The increase of D. salina from 25% to 50% in the co-digestion mixture clearly increased the biodegradability of the sole substrates. The highest biodegradability was found for the co-digestion mixture 50% OMSW-50% D. salina. Nevertheless, the maximum methane production, 330mLCH4/gVSadded, and the highest methane production rate were obtained for the co-digestion mixture 75% OMSW-25% D. salina, keeping a C/N ratio near to 26.7/1.

Transcription of fdh and hyd in Syntrophobacter spp. and Methanospirillum spp. as a diagnostic tool for monitoring anaerobic sludge deprived of molybdenum, tungsten and selenium

Formate dehydrogenases and hydrogenases contain molybdenum or tungsten and/or selenium. These enzymes are crucial for interspecies formate and hydrogen transfer between propionate degrading Syntrophobacter spp. and methanogenic Methanospirillum spp. Here we used reverse transcription of total RNA followed by quantitative PCR (RT-qPCR) with specific primers to get insight into interspecies formate and hydrogen transfer. Transcriptional regulation of formate dehydrogenases and hydrogenases in Syntrophobacter and Methanospirillum spp. in a propionate-fed up-flow anaerobic sludge bed (UASB) reactor was examined. In both microorganisms formate dehydrogenase and hydrogenase coding genes (fdh and hyd respectively) were transcribed simultaneously. During 249 days in which molybdenum, tungsten and selenium were not supplied to the reactor feed, the microbial activity and transcription of fdh and hyd in Syntrophobacter spp. decreased. Transcription of fdh and hyd in Methanospirillum spp. did not decrease, but transcription of fdh increased when after 249 days molybdenum, tungsten and selenium were supplied to the reactor feed. The developed RT-qPCR is a technique that can give rapid information about active processes in methanogenic granular sludge and may contribute to predict metal limitation and failure in UASB reactors.

Batch anaerobic co-digestion of waste activated sludge and microalgae (Chlorella sorokiniana) at mesophilic temperature

ABSTRACT The microalgae Chlorella sorokiniana are used as co-substrate for waste activated sludge (WAS) anaerobic digestion. The specific objective of this research was to evaluate the feasibility of improving methane production from anaerobic digestion of WAS in co-digestion with this microalga, based on an optimized mixture percentage. Thus, the anaerobic co-digestion of both substrates aims to overcome the drawbacks of the anaerobic digestion of single WAS, simultaneously improving its management. Different co-digestion mixtures (0% WAS-100% microalgae; 25% WAS-75% microalgae; 50% WAS-50% microalgae; 75% WAS-25% microalgae; 100% WAS-0% microalgae) were studied. The highest methane yield (442 mL CH4/g VS) was obtained for the mixture with 75% WAS and 25% microalgae. This value was 22% and 39% higher than that obtained in the anaerobic digestion of the sole substrates WAS and microalgae, respectively, as well as 16% and 25% higher than those obtained for the co-digestion mixtures with 25% WAS and 75% microalgae and 50% WAS and 50% microalgae, respectively. The kinetic constant of the process increased 42%, 42% and 12%, respectively, for the mixtures with 25%, 50% and 75% of WAS compared to the substrate without WAS. Anaerobic digestion of WAS, together with C. sorokiniana, has been clearly improved by ensuring its viability, suitability and efficiency.

Influence of an oxic settling anoxic system on biomass yield, protozoa and filamentous bacteria

An oxic settling anoxic system coupled with an activated sludge process has been studied to reduce sewage sludge production. The reduction of sludge yield, excess sludge production and active biomass yield were 51.7%, 52.9% and 67.1%, respectively, compared with the control system. The oxic reactor of the oxic settling anoxic system, even with a lower active biomass concentration than the oxic reactor of control system, showed a higher metabolic activity in their active biomass. Diversity and crawling ciliates group have been shown as promising bioindicators of active biomass yield reduction. The identification of floc-forming bacteria in the control system suggested that oxic settling anoxic system will improve settling properties compared to a Conventional Activated Sludge process.

Fate of Trace Metals in Anaerobic Digestion

A challenging, and largely uncharted, area of research in the field of anaerobic digestion science and technology is in understanding the roles of trace metals in enabling biogas production. This is a major knowledge gap and a multifaceted problem involving metal chemistry; physical interactions of metal and solids; microbiology; and technology optimization. Moreover, the fate of trace metals, and the chemical speciation and transport of trace metals in environments--often agricultural lands receiving discharge waters from anaerobic digestion processes--simultaneously represents challenges for environmental protection and opportunities to close process loops in anaerobic digestion.

Challenges for Cost-Effective Microalgae Anaerobic Digestion

1. Microalgae photosynthesis allows biological CO2 fixation, which is expected to mitigate atmospheric CO2 increase (Amin 2009; Brennan & Owende 2010; Mutanda et al. 2011). 2. Microalgae are 10 – 50 times more efficient than plants in terms of CO2 fixation (Wang et al. 2008). Thus, microalgae can fix 1.83 tonnes of CO2 per 1 tonne of produced microalgae (Chisti 2007). 3. Microalgae can be produced on non-arable areas such as lakes, oceans or deserts, thus reducing competition with food production (Mussgnug et al. 2010; Stephens et al. 2010). This advantage is a key factor when energy supply is considered in desert zones near oceans. 4. Some microalgae can grow under saline conditions, which strengthen the use of micro‐ algae as feedstock for biofuel production in desert zones near the ocean when freshwater supply is not feasible.

Effect of sorption kinetics on nickel toxicity in methanogenic granular sludge

This study investigates the effect of nickel speciation and its equilibrium kinetics on the nickel toxicity to methylotrophic methanogenic activity. Toxicity tests were done with anaerobic granular sludge in three different media containing variable concentrations of complexing ligands. A correlation between nickel toxicity and the free nickel concentration failed, because not the equilibrium conditions, but the kinetics of the speciation processes taking place in the medium (precipitation, sorption, liquid speciation, etc.) determine nickel bio-uptake and its toxic effect. The latter was confirmed with an F-test (p-value always lower than 0.1). It was shown that the biological activity (methane production) took place within 3-20 days upon the start of methanogenic experiments, i.e. prior the chemical-physical equilibrium of nickel speciation was established in the methanogenic medium (10-20 days). The process of nickel sorption in the methanogenic granular sludge was limited by intra-particle diffusion and the experimental data fitted to the Weber-Morris sorption model. The other sorption kinetic models applied (pseudo-first order sorption kinetics, pseudo-second order sorption kinetics and first order reversible reaction kinetics) did not fit the experimental data satisfactorily.