Fertirrigation with sugarcane vinasse: Foreseeing potential impacts on soil and water resources through vinasse characterization.

This paper reports the characterization of the polluting potential of sugarcane vinasse, the main wastewater from ethanol production. Compositional data from vinasse samples collected from sugarcane biorefineries were used to predict negative effects on the soil, water resources and crops potentially associated with fertirrigation, the primary final destination of vinasse in Brazil. High risks of soil salinization were associated with the land disposal of vinasse, as evidenced by the high levels of total dissolved solids (TDS; >4,000 mg L-1) and electrical conductivity (>6.7 dS m-1). The high TDS levels coupled with the high biodegradable organic content of vinasse (>14 g L-1) also favor organic overloading events, leading to local anaerobiosis conditions. Conversely, soil sodification should not be observed in areas fertirrigated with sugarcane vinasse, given the low Na concentrations (<66 mg L-1) relative to Mg (>145.1 mg L-1) and Ca (>458.4 mg L-1) levels. Priority pollutants (Cu, Cr, Ni, Pb and Zn) and phytotoxic elements (Al and Fe) were also found in the analyzed samples; however, relevant environmental impacts should not be associated with these particular constituents. Overall, the relatively simple methodology used herein could efficiently replace massive field data collection to provide a basic understanding of the fate of vinasse in the environment in order to highlight the priority points to be considered in the management of this effluent. In summary, the prompt implementation of treatment plants in distilleries, in addition to a continuous and broad compositional characterization of vinasse, is essential to guarantee its adequate reuse.

Microbial community dynamics and stability during an ammonia-induced shift to syntrophic acetate oxidation.

Anaerobic digesters rely on the diversity and distribution of parallel metabolic pathways mediated by complex syntrophic microbial communities to maintain robust and optimal performance. Using mesophilic swine waste digesters, we experimented with increased ammonia loading to induce a shift from aceticlastic methanogenesis to an alternative acetate-consuming pathway of syntrophic acetate oxidation. In comparison with control digesters, we observed shifts in bacterial 16S rRNA gene content and in functional gene repertoires over the course of the digesters' 3-year operating period. During the first year, under identical startup conditions, all bioreactors mirrored each other closely in terms of bacterial phylotype content, phylogenetic structure, and evenness. When we perturbed the digesters by increasing the ammonia concentration or temperature, the distribution of bacterial phylotypes became more uneven, followed by a return to more even communities once syntrophic acetate oxidation had allowed the experimental bioreactors to regain stable operation. The emergence of syntrophic acetate oxidation coincided with a partial shift from aceticlastic to hydrogenotrophic methanogens. Our 16S rRNA gene analysis also revealed that acetate-fed enrichment experiments resulted in communities that did not represent the bioreactor community. Analysis of shotgun sequencing of community DNA suggests that syntrophic acetate oxidation was carried out by a heterogeneous community rather than by a specific keystone population with representatives of enriched cultures with this metabolic capacity.

Bacterial community structures are unique and resilient in full-scale bioenergy systems.

Anaerobic digestion is the most successful bioenergy technology worldwide with, at its core, undefined microbial communities that have poorly understood dynamics. Here, we investigated the relationships of bacterial community structure (>400,000 16S rRNA gene sequences for 112 samples) with function (i.e., bioreactor performance) and environment (i.e., operating conditions) in a yearlong monthly time series of nine full-scale bioreactor facilities treating brewery wastewater (>20,000 measurements). Each of the nine facilities had a unique community structure with an unprecedented level of stability. Using machine learning, we identified a small subset of operational taxonomic units (OTUs; 145 out of 4,962), which predicted the location of the facility of origin for almost every sample (96.4% accuracy). Of these 145 OTUs, syntrophic bacteria were systematically overrepresented, demonstrating that syntrophs rebounded following disturbances. This indicates that resilience, rather than dynamic competition, played an important role in maintaining the necessary syntrophic populations. In addition, we explained the observed phylogenetic differences between all samples on the basis of a subset of environmental gradients (using constrained ordination) and found stronger relationships between community structure and its function rather than its environment. These relationships were strongest for two performance variables--methanogenic activity and substrate removal efficiency--both of which were also affected by microbial ecology because these variables were correlated with community evenness (at any given time) and variability in phylogenetic structure (over time), respectively. Thus, we quantified relationships between community structure and function, which opens the door to engineer communities with superior functions.

Interaction between temperature and ammonia in mesophilic digesters for animal waste treatment.

Four anaerobic sequencing batch reactors (ASBRs) were operated during a period of 988 days to evaluate the effect of temperature, ammonia, and their interconnectivity on the methane yield of anaerobic processes for animal waste treatment. During period 1 (day 0-378), the methane yield was 0.31 L CH(4)/g volatile solids (VS) for all digesters (with no statistical differences among them) at a temperature and total ammonium-N levels of 25 degrees C and approximately 1200 mg NH(4)(+)-N/L, respectively. During period 2 (day 379-745), the methane yield at 25 degrees C decreased by 45% when total ammonium-N and ammonia-N were increased in two of the four ASBRs to levels >4000 mg NH(4)(+)-N/L and >80 mg NH(3)-N/L, respectively. During period 3 (day 746-988), this relative inhibition was reduced from 45% to 13% compared to the low-ammonia control reactors when the operating temperature was increased from 25 degrees C to 35 degrees C (while the free ammonia levels increased from approximately 100 to approximately 250 mg NH(3)-N/L). The 10 degrees C increase in temperature doubled the rate constant for methanogenesis, which overwhelmed the elevated toxicity effects caused by the increasing concentration of free ammonia. Thus, the farmer/operator may alleviate ammonia toxicity by increasing the operating temperature within the mesophilic range. We extrapolated our data to correlate temperature, ammonia, and methane yield and to hypothesize that the difference between high- and low-ammonia reactors is negligible at the optimum mesophilic temperature of 38 degrees C.

Effect of shear on performance and microbial ecology of continuously stirred anaerobic digesters treating animal manure.

We determined the effect of different mixing intensities on the performance, methanogenic population dynamics, and juxtaposition of syntrophic microbes in anaerobic digesters treating cow manure from a dairy farm. Computer automated radioactive particle tracking in conjunction with computational fluid dynamics was performed to quantify the shear levels locally. Four continuously stirred anaerobic digesters were operated at different mixing intensities of 1,500, 500, 250, and 50 revolutions per min (RPM) over a 260-day period at a temperature of 34 +/- 1 degrees C. Animal manure at a volatile solids (VS) concentration of 50 g/L was fed into the digesters daily at five different organic loading rates between 0.6 and 3.5 g VS/L day. The different mixing intensities had no effect on the biogas production rates and yields at steady-state conditions. A methane yield of 0.241 +/- 0.007 L CH(4)/g VS fed was obtained by pooling the data of all four digesters during steady-state periods. However, digester performance was affected negatively by mixing intensity during startup of the digesters, with lower biogas production rates and higher volatile fatty acids concentrations observed for the 1,500-RPM digester. Despite similar methane production yields and rates, the acetoclastic methanogenic populations were different for the high- and low-intensity mixed digesters with Methanosarcina spp. and Methanosaeta concilii as the predominant methanogens, respectively. For all four digesters, epifluorescence microscopy revealed decreasing microbial floc sizes beginning at week 4 and continuing through week 26 after which no microbial flocs remained. This decrease in size, and subsequent loss of microbial flocs did not, however, produce any long-term upsets in digester performance.

Anaerobic digestion of secondary residuals from an anaerobic bioreactor at a brewery to enhance bioenergy generation.

Many beer breweries use high-rate anaerobic digestion (AD) systems to treat their soluble high-strength wastewater. Biogas from these AD systems is used to offset nonrenewable energy utilization in the brewery. With increasing nonrenewable energy costs, interest has mounted to also digest secondary residuals from the high-rate digester effluent, which consists of yeast cells, bacteria, methanogens, and small (hemi)cellulosic particles. Mesophilic (37 degrees C) and thermophilic (55 degrees C) lab-scale, low-rate continuously-stirred anaerobic digestion (CSAD) bioreactors were operated for 258 days by feeding secondary residuals at a volatile solids (VS) concentration of approximately 40 g l(-1). At a hydraulic retention time (HRT) of 15 days and a VS loading rate of 2.7 g VS l(-1) day(-1), the mesophilic bioreactor showed an average specific volumetric biogas production rate of 0.88 l CH4 l(-1) day(-1) and an effluent VS concentration of 22.2 g VS l(-1) (43.0% VS removal efficiency) while the thermophilic bioreactor displayed similar performances. The overall methane yield for both systems was 0.21 l CH4 g(-1) VS fed and 0.47-0.48 l CH4 g(-1) VS removed. A primary limitation of thermophilic digestion of this protein-rich waste is the inhibition of methanogens due to higher nondissociated (free) ammonia (NH3) concentrations under similar total ammonium (NH4+) concentrations at equilibrium. Since thermophilic AD did not result in advantageous methane production rates or yields, mesophilic AD was, therefore, superior in treating secondary residuals from high-rate AD effluent. An additional digester to convert secondary residuals to methane may increase the total biogas generation at the brewery by 8% compared to just conventional high-rate digestion of brewery wastewater alone.

Thermophilic anaerobic digestion to increase the net energy balance of corn grain ethanol.

U.S. production of fuel ethanol from corn grain has increased considerably over the last 10 years. Intense debate regarding the true environmental impact of the overall production process has been ongoing. The present study evaluated the utilization of thin stillage (a major byproduct of the dry-mill corn grain-to-ethanol process) in laboratory-scale thermophilic anaerobic sequencing batch reactors for conversion to methane. We found that augmentation of cobalt as a growth factor to the thermophilic anaerobic digestion process is required. After reaching sustainable operating performances, the methane potential in the reactors was 0.254 L CH4/g total chemical oxygen demand (TCOD) fed. Together with a reduction in the mass of solids that needs drying, methane generation translates to a 51% reduction of natural gas consumption at a conventional dry mill, which improves the net energy balance ratio from 1.26 to 1.70. At the design hydraulic retention time of 10 days, the digesters achieved TCOD, biodegradable COD, volatile solids, and total solids removal efficiencies of 90%, 75%, 89%, and 81%, respectively. We also found that struvite precipitation occurred in the thermophilic digesters during the course of the study, resulting in possibilities for nutrient recovery.

A portable anaerobic microbioreactor reveals optimum growth conditions for the methanogen Methanosaeta concilii.

Conventional studies of the optimum growth conditions for methanogens (methane-producing, obligate anaerobic archaea) are typically conducted with serum bottles or bioreactors. The use of microfluidics to culture methanogens allows direct microscopic observations of the time-integrated response of growth. Here, we developed a microbioreactor (microBR) with approximately 1-microl microchannels to study some optimum growth conditions for the methanogen Methanosaeta concilii. The microBR is contained in an anaerobic chamber specifically designed to place it directly onto an inverted light microscope stage while maintaining a N2-CO2 environment. The methanogen was cultured for months inside microchannels of different widths. Channel width was manipulated to create various fluid velocities, allowing the direct study of the behavior and responses of M. concilii to various shear stresses and revealing an optimum shear level of approximately 20 to 35 microPa. Gradients in a single microchannel were then used to find an optimum pH level of 7.6 and an optimum total NH4-N concentration of less than 1,100 mg/liter (<47 mg/liter as free NH3-N) for M. concilii under conditions of the previously determined ideal shear stress and pH and at a temperature of 35 degrees C.