Impact of Bacillus subtilis on manure solids, odor, and microbiome.

A study was conducted to determine the effectiveness of supplementing swine manure with Bacillus subtilis (BS) to improve digestion of manure solids and lower odor emission. Large bioreactors (400 L) with manure (100 L) were treated with commercially available BS at a rate of 1% manure volume by either directly pouring or surface spraying the manure with inoculum. Manure physicochemical properties, gas emissions, and microbiome were monitored. Manures treated multiple times with BS or surface sprayed had significantly (P < 0.05) lower electrical conductivity, volatile solids, and chemical oxygen demand, by 3-5% compared to non-treated control manures. Volatile sulfur compound emissions (VSCs) were reduced by 20-30% in both experiments, while ammonia and volatile organic compounds were reduced by 40% and 15%, respectively, in surface spray experiment only. The manure indigenous microbiome remained relatively stable following treatment and BS were never detected in the raw or treated manure following multiple treatments. The reduction in manure organic carbon and VSCs emissions were a result of physical mixing during manure treatment and biological material in the microbial inoculum stimulating microbial activity and not growth of BS.

Comprehensive analysis of microbial dynamics linked with the reduction of odorous compounds in a full-scale swine manure pit recharge system with recirculation of aerobically treated liquid fertilizer.

It is believed that the generation of odorous materials in manure-slurry pits during the storage can be reduced by recirculating aerobically treated liquid fertilizer (ATLF) to a manure-pit recharge system (PRS). However, the biological mechanisms for reduction of those problematic compounds remain poorly understood. In this study, the links between microbial evolution and changes in chemical composition and odorous compounds were analyzed where swine-manure slurry was stored in a full-scale PRS. Some beneficial microorganisms were successfully established in the PRS. This resulted in the accumulation of fewer undesirable chemical components and lower amounts of odorous compounds compared to those in a conventional swine-manure slurry pit (the control). Decrease in the volatile fatty acids (1387-8478 mg/L â†’ 306-1258 mg/L) and NH3 (3387-4300 mg/L â†’ 85-200 mg/L) in the PRS was mainly due to the development of a key community that included a mix of aerobic, anaerobic fermentative, nitrifying (0.1-0.6%) and denitrifying (1.7-3.5%), and methanogenic microorganisms (2.1-4.2%). Meanwhile, the generation of greater amounts of H2S (12-290 mg/L â†’ 61-1754 mg/L) was found in the PRS, which condition was supported by the increased proportion of sulfate-reducing bacteria (0.5-3%). To the authors' best knowledge this is the first study comprehensively analyzing microbial dynamics linked with the reduction of odorous compounds in the full-scale PRS in response to recirculation of ATLF.

Comprehensive analysis of the microbial communities and operational parameters of two full-scale anaerobic digestion plants treating food waste in South Korea: Seasonal variation and effect of ammonia.

There are about ninety full-scale anaerobic digestion (AD) plants in South Korea that treat food waste (FW); however, the key diff ;erences in the microbial communities in different seasons and the effects of ammonia in AD remain poorly understood. In this study, the seasonal changes in microbial communities associated with operational parameters of two full-scale ADs (C and W plants) treating FW were analyzed. The organic loading rate (OLR) variability had an influence on the seasonal CH4 yield; the W plant had a lower CH4 yield with an unstable AD performance while the C plant had a higher CH4 yield with a stable AD performance. It was mainly due to the substantially different NH4+ concentration; the W plant had a NH4+ concentration nearly 1.6 times higher compared to the C plant. The high NH4+ presence in the W plant led to the dominance of class Clostridia, and methanogenesis was mostly done by hydrogenotrophs (Methanomassiliicoccus luminyensis). Additionally, the members belonging to Clostridia and Bacteroidia were found at both plants in each season (share ≥0.5%) implying their indispensable role during the anaerobic digestion of FW.

Selective removal of color substances by carbon-based adsorbents in livestock wastewater effluents.

Livestock wastewater effluent generated after the anaerobic treatment process contains the considerable amount of color-causing organic matter. In this study, a quantitative comparison of three carbon-based adsorbents included granular activated carbon (GAC), expanded graphite (EG), and multi-walled carbon nanotubes (MWNTs) was carried out for the potential application to the removal of color substances, and their mechanism was proposed. Although GAC showed the highest specific dissolved organic carbon (DOC) adsorption capacity, the color removal efficiency was the smallest among three adsorbents. The selective color removal ratios of EG and MWNTs reached 22.7 ± 0.1 PtCo/mg-DOC-removed and 21.2 ± 0.1 PtCo/mg-DOC-removed, respectively, while that of GAC was only 12.3 ± 0.1 PtCo/mg-DOC-removed. The selective adsorption of color substances by graphene-based carbon materials was due to the aromatic π-π interaction between organic matter and the hexagonal carbon lattice of graphene. The analysis of molecular weight distribution also confirmed that the exposed surface area and macro-pores were responsible for the adsorption of high molecular weight color substances. The chemical regeneration of three adsorbents was examined using 1% NaOCl solution and MWNTs showed almost complete recovery of the initial color removal capacity. In conclusion, MWNTs were the most suitable carbon nanomaterial for the selective color removal from livestock wastewater effluent.

Preparation of Highly Porous PAN-LATP Membranes as Separators for Lithium Ion Batteries.

Separators are a vital component to ensure the safety of lithium-ion batteries. However, the commercial separators employed in lithium ion batteries are inefficient due to their low porosity. In the present study, a simple electrospinning technique is adopted to prepare highly porous polyacrylonitrile (PAN)-based membranes with a higher concentration of lithium aluminum titanium phosphate (LATP) ceramic particles, as a viable alternative to the commercialized separators used in lithium ion batteries. The effect of the LATP particles on the morphology of the porous membranes is demonstrated through Field emission scattering electron microscopy. X-ray diffraction and Fourier transform infrared spectra studies suitably demonstrate the mixing of PAN and LATP particles in the polymer matrix. PAN with 30 wt% LATP (P-L30) exhibits an enhanced porosity of 90% and is more thermally stable, with the highest electrolyte uptake among all the prepared membranes. Due to better electrolyte uptake, the P-L30 membrane demonstrates an improved ionic conductivity of 1.7 mS/cm. A coin cell prepared with a P-L30 membrane and a LiFePO4 cathode demonstrates the highest discharge capacity of 158 mAh/g at 0.5 C-rate. The coin cell with the P-L30 membrane also displays good cycling stability by retaining 97.5% of the initial discharge capacity after 200 cycles of charging and discharging at a 1C rate.

Sulfate reducing bacteria-based wastewater treatment system integrated with sulfide fuel cell for simultaneous wastewater treatment and electricity generation.

This study aimed to design a sulfate-reducing bacteria (SRB)-based wastewater treatment system (SWTS) integrated with a sulfide fuel cell (SFC) as an alternative to the energy-intensive aerobic wastewater treatment process. The result showed that the COD/sulfate ratio and hydraulic retention time (HRT) were two important parameters in a SWTS. The highest COD and sulfate removal efficiency rates were at a HRT of 4 h at a COD/sulfate ratio of 0.67, reaching 83 ±â€¯0.2% and 84 ±â€¯0.4% with sulfate removal rates of 4.087 ±â€¯32 mg SO42-/d, respectively. A microbial analysis revealed that the dominance of nine OTUs belonging to SRB closely affected the high sulfate removal efficiency in the SWTS. At the HRT of 8 h, voltage of 0.02 V and a power density level of 130 mW/m2 were obtained with sulfide removal efficiency of 99 ±â€¯0.5%. These results overall demonstrate that SRB can serve as a green and effective route for wastewater treatment.

Effects of pig slurry acidification on methane emissions during storage and subsequent biogas production.

In addition to undesirable odorous gases, substantial amounts of greenhouse gases (GHG), particularly methane (CH4), are generated during the storage of livestock manure. To reduce the CH4 emissions, first, pig slurry (PS) was stored for 40 d at 30 °C after adjusting the pH at 5.0-7.0 using H2SO4 solution. In the control (non-acidified PS), 3.7 kg CO2 eq./ton PS of CH4 emissions was detected, which was reduced to 1.8, 0.9, 0.4, 0.2, and 0.1 kg CO2 eq./ton PS at pH 7.0, 6.5, 6.0, 5.5, and 5.0, respectively. Methanosarcina was found to be the dominant genus (67% of the total archaeal sequence) in the control, whose dominance was reduced as storage pH decreased. The results of ribonucleic acid analysis and specific methanogenic activity test further confirmed the inhibition of indigenous methanogens by acidification. Later, the biochemical CH4 potential of stored PS was tested. Compared to the control (10.6 L CH4/L PS), the acidified PS showed higher CH4 yields of 12.7-14.6 L CH4/L PS, presumably by keeping degradable organic matters in PS under acidic condition. Among different acidification pHs tested, the maximum amount of GHG reduction was achieved at pH 6.0 by reducing CH4 emission to +0.4 kg CO2 eq./ton PS during storage while increasing biogas production potential equivalent to 48.3 kWh/ton PS (-22.5 kg CO2 eq./ton PS), resulting in a further reduction of (-)9.6 kg CO2 eq./ton PS compared to the control.

Increased biodegradability of low-grade coal wastewater in anaerobic membrane bioreactor by adding yeast wastes.

Demineralization is required in upgrading low-grade coal to serve as an alternative energy resource for the production of fuel and valuable chemicals but generates a large amount of low-grade coal wastewater (LCWW). The objective of this study was to investigate the effects of a co-substrate on an anaerobic membrane bioreactor (AnMBR) treating LCWW. CH4 was not produced during the operation fed by LCWW alone. When yeast wastes (YW) were supplemented, there was a gradual increase in the biodegradability of LCWW, achieving 182 CH4 mL/g COD with 58% COD removal efficiency. The analysis of physicochemical characteristics in the effluent of AnMBR, done by excitation-emission matrix (EEM) and size exclusion chromatography (SEC), showed that the proportion of soluble microbial products (SMPs) and aromatic group with high-molecular weight (>1 kDa) increased. Microbial analysis revealed that the increased dominance of bacteria Comamonas, Methanococcus, and Methanosarcina facilitated biodegradation of LCWW in the presence of YW.

High-calorific bio-hydrogen production under self-generated high-pressure condition.

For the use of biologically produced H2, removal of CO2 is an indispensable process. Unlike conventional CO2 removal methods, this study proposed a self-generated high-pressure dark fermentation (HPDF) process as a novel strategy for directly producing high-calorific bio-H2. The pressure was automatically increased by self-generated gas, while the maximum pressure inside fermenter was restricted to 1, 3, 5, 7, and 10 bar in a batch operation. As the pressure increased from 1 to 10 bar, the H2 content increased from 55% to 80%, whereas the H2 yield decreased from 1.5 to 0.9 mol H2/mol hexoseadded. The highest H2 content of 80% was obtained at both of 7 and 10 bars. Increased lactate production with increased abundance of lactic acid bacteria was observed at high-pressure. Despite the lower H2 yields at high-pressure conditions, HPDF was found to be economically beneficial for obtaining high-calorific bio-H2 owing to the low CO2 removal cost.

Biohydrogen production from food waste: Current status, limitations, and future perspectives.

Among the various biological routes for H2 production, dark fermentation is considered the most practically applicable owing to its capability to degrade organic wastes and high H2 production rate. Food waste (FW) has high carbohydrate content and easily hydrolysable in nature, exhibiting higher H2 production potential than that of other organic wastes. In this review article, first, the current status of H2 production from FW by dark fermentation and the strategies applied for enhanced performance are briefly summarized. Then, the technical and economic limitations of dark fermentation of FW are thoroughly discussed. Economic assessment revealed that the economic feasibility of H2 production from FW by dark fermentation is questionable. Current efforts to further increase H2 yield and waste removal efficiency are also introduced. Finally, future perspectives along with possible routes converting dark fermentation effluent to valuable fuels and chemicals are discussed.

Effect of feeding mode and dilution on the performance and microbial community population in anaerobic digestion of food waste.

The effect of feeding mode and dilution was studied in anaerobic digestion of food waste. An upflow anaerobic digester with a settler was fed at six different organic loading rates (OLRs) from 4.6 to 8.6kgCOD/m3/d for 200days. The highest methane productivity of 2.78LCH4/L/d was achieved at 8.6kgCOD/m3/d during continuous feeding of diluted FW. Continuous feeding of diluted food waste showed more stable and efficient performance than stepwise feeding of undiluted food waste. Sharp increase in propionate concentration attributed towards deterioration of the digester performances in stepwise feeding of undiluted food waste. Microbial communities at various OLRs divulged that the microbial distribution in the continuous feeding of diluted food waste was not significantly perturbed despite the increase of OLR up to 8.6kgCOD/m3/d, which was contrast to the unstable distribution in stepwise feeding of undiluted food waste at 6.1kgCOD/m3/d.

Low-strength ultrasonication positively affects methanogenic granules toward higher AD performance: Hydrolytic enzyme excretions.

In our previous studies, enhanced methane (CH4) production using low-strength ultrasonication was achieved and the results were evidenced by physico-chemical and molecular biological approaches. As a final continuation study, the effects of low-strength ultrasonication on the activities of hydrolytic enzymes (amylase, cellulase, and protease) were investigated on methanogenic granules given that hydrolysis regulates the whole anaerobic digestion (AD) process. Up to 213% enhanced hydrolytic enzyme activities were observed, and they seem to be highly related to the enhanced CH4 production. However, the effects of ultrasonication on the distribution (liquid- and solid-phases) of hydrolytic enzymes were negligible. Enzymatic activation by low-strength ultrasonication was hypothetically caused by acoustic streaming, presumably enabling to overcome the masking effect, substrate inhibition and spatial constraint.

Cultivation of four microalgae species in the effluent of anaerobic digester for biodiesel production.

This study investigated if an effluent from anaerobic digestion (AD) system can be used as a nutrients source for the microalgae cultivation, and in so doing, if the effluent can be properly treated. Nitrogen and phosphorus in the AD effluent well supported microalgal growth, and their removal efficiency reached >97.9% and 99.2%, respectively. Among four different algal species tested, Micractinium inermum particularly stood out, showing the highest biomass and FAME productivity: 0.16gL-1d-1 with 3.23gL-1 of dry cell weight, and 0.04gL-1d-1 with 27.54% (w/w) of FAME contents, respectively. As the concentrations of the nutrients decreased over time, the FAME contents were increased and its quality as well, satisfying several biodiesel quality standards. This study supports that the AD effluent can indeed serve as a cheap and nutrient-rich medium for microalgae cultivation, and equally importantly, microalgae can be a workable treatment option for it.

Producing desulfurized biogas through removal of sulfate in the first-stage of a two-stage anaerobic digestion.

In the present work, a two-stage anaerobic digestion system (TSADS) was newly designed to produce biogas with a greatly reduced H2 S content. The role of first (sulfidogenic)-stage digester was not only acidogenesis but also sulfidogenesis (sulfate reduction to H2 S), which would minimize the input of H2 S-producing source in the followed second (methanogenic)-stage digester. For the coexistence of acidogens and sulfate reducing bacteria (SRB) in the sulfidogenic-stage digester, it was found that pH played a crucial role. The acidogenic activity was not affected within a pH range of 4.5-6.0, while it was important to maintain a pH at 5.5 to achieve a sulfate removal efficiency over 70%. The highest sulfate removal attained was 78% at a hydraulic retention time (HRT) of 5 h at pH 5.5 ± 0.1. The H2 S content in the biogas produced in the conventional single-stage digester (SSAD), used as a control, reached 1,650 ± 25 ppmv . In contrast, the biogas produced in the methanogenic-stage digester of the developed process had an H2 S content of 200 ± 15 ppmv . Microbial analysis, done by the next generation sequencing technique, clearly showed the changes in community under different operating conditions. Desulfovibrio bastinii (4.9%) played a key role in sulfate removal in the sulfidogenic-stage of the TSADS owing to its characteristics of a short doubling time and growth in an acidic environment. Biotechnol. Bioeng. 2017;114: 970-979. © 2016 Wiley Periodicals, Inc.

More value from food waste: Lactic acid and biogas recovery.

Anaerobic digestion (AD) is one of the traditional technologies for treating organic solid wastes, but its economic benefit is sometimes questioned. To increase the economic feasibility of the treatment process, the aim of this study was to recover not only biogas from food waste but lactic acid (LA) as well. At first, LA fermentation of food waste (FW) was conducted using an indigenous mixed culture. During the operation, temperature was gradually increased from 35 °C to 55 °C, with the highest performance attained at 50 °C. At 50 °C and hydraulic retention time (HRT) of 1.0 d, LA concentration in the broth was 40 kg LA/m(3), corresponding to a yield of 1.6 mol LA/mol hexoseadded. Pyrosequencing results showed that Lactobacillus (97.6% of the total number of sequences) was the predominant species performing LA fermentation of FW. The fermented broth was then centrifuged and LA was extracted from the supernatant by the combined process of nanofiltration and water-splitting electrodialysis. The process could recover highly purified LA by removing 85% of mineral ions such as Na(+), K(+), Mg(2+), and Ca(2+) and 90% of residual carbohydrates. Meanwhile, the solid residue remained after centrifugation was further fermented to biogas by AD. At HRT 40 d (organic loading rate of 7 kg COD/m(3)/d), the highest volumetric biogas production rate of 3.5 m(3)/m(3)/d was achieved with a CH4 yield of 0.25 m(3) CH4/kg COD. The mass flow showed that 47 kg of LA and 54 m(3) of biogas could be recovered by the developed process from 1 ton of FW with COD removal efficiency of 70%. These products have a higher economic value 60 USD/ton FW compared to that of conventional AD (27 USD/ton FW).

Mitigation of ammonia inhibition by internal dilution in high-rate anaerobic digestion of food waste leachate and evidences of microbial community response.

A high-rate anaerobic digestion of food waste leachate were tested using intermittent continuously stirred tank reactors (iCSTRs) to evaluate how severe ammonia inhibition could be mitigated with internal dilution strategy, and to identify how bacterial and archaeal community respond in genus and species level. Experimental results show that the digestion performance was well maintained up to hydraulic retention time (HRT) of 40 days but could not keep steady-state as HRT decreased to 30 days due to severe free ammonia (FA) inhibition. Coupling internal dilution was the key to relieve the inhibition since it reduced FA concentration as low as 62 mg/L even at HRT 30 days, which corresponds to organic loading rate of 5 g COD/L/d, demonstrating CH4 yield of 0.32 L CH4 /g CODadded . It was confirmed that the dilution offers iCTSRs manage severe ammonia inhibition with the balanced community structure between bacteria and archaea in this high-rate anaerobic digestion. Genus and species level pyrosequencing evidence that FA inhibition to community dynamics of Methanosarcina and Methanosaeta is strongly connected to methanogenesis, and Methanosarcina plays a key role in an iCSTR with the dilution. Biotechnol. Bioeng. 2016;113: 1892-1901. © 2016 Wiley Periodicals, Inc.

Microbial granulation for lactic acid production.

This work investigated the formation of microbial granules to boost the productivity of lactic acid (LA). The flocculated form of LA-producing microbial consortium, dominated by Lactobacillus sp. (91.5% of total sequence), was initially obtained in a continuous stirred-tank reactor (CSTR), which was fed with 2% glucose and operated at a hydraulic retention time (HRT) of 12 h and pH 5.0 ± 0.1 under a thermophilic condition (50°C). The mixed liquor in the CSTR was then transferred to an up-flow anaerobic sludge blanket reactor (UASB). The fermentation performance and granulation process were monitored with a gradual decrease of HRT from 8.0 to 0.17 h, corresponding to an increase in the substrate loading from 60 to 2,880 g glucose L(-1) d(-1) . As the operation continued, the accumulation of biomass in the UASB was clearly observed, which changed from flocculent to granular form with decrease in HRT. Up to the HRT decrease to 0.5 h, the LA concentration was maintained at 19-20 g L(-1) with over 90% of substrate removal efficiency. However, further decrease of HRT resulted in a decrease of LA concentration with increase in residual glucose. Nevertheless, the volumetric LA productivity continuously increased, reaching 67 g L-fermenter (-1) h(-1) at HRT 0.17 h. The size of LA-producing granules and hydrophobicity gradually increased with decrease in HRT, reaching 6.0 mm and 60%, respectively. These biogranules were also found to have high settling velocities and low porosities, ranging 2.69-4.73 cm s(-1) and 0.39-0.92, respectively.

Inhibition of residual n-hexane in anaerobic digestion of lipid-extracted microalgal wastes and microbial community shift.

Converting lipid-extracted microalgal wastes to methane (CH4) via anaerobic digestion (AD) has the potential to make microalgae-based biodiesel platform more sustainable. However, it is apparent that remaining n-hexane (C6H14) from lipid extraction could inhibit metabolic pathway of methanogens. To test an inhibitory influence of residual n-hexane, this study conducted a series of batch AD by mixing lipid-extracted Chlorella vulgaris with a wide range of n-hexane concentration (∼10 g chemical oxygen demand (COD)/L). Experimental results show that the inhibition of n-hexane on CH4 yield was negligible up to 2 g COD/L and inhibition to methanogenesis became significant when it was higher than 4 g COD/L based on quantitative mass balance. Inhibition threshold was about 4 g COD/L of n-hexane. Analytical result of microbial community profile revealed that dominance of alkane-degrading sulfate-reducing bacteria (SRB) and syntrophic bacteria increased, while that of methanogens sharply dropped as n-hexane concentration increased. These findings offer a useful guideline of threshold n-hexane concentration and microbial community shift for the AD of lipid-extracted microalgal wastes.

Scenedesmus-based treatment of nitrogen and phosphorus from effluent of anaerobic digester and bio-oil production.

In this study, a microalgae-based technology was employed to treat wastewater and produce biodiesel at the same time. A local isolate Scenedesmus sp. was found to be a well suited species, particularly for an effluent from anaerobic digester (AD) containing low carbon but high nutrients (NH3-N=273mgL(-1), total P=58.75mgL(-1)). This algae-based treatment was quite effective: nutrient removal efficiencies were over 99.19% for nitrogen and 98.01% for phosphorus. Regarding the biodiesel production, FAME contents of Scenedesmus sp. were found to be relatively low (8.74% (w/w)), but overall FAME productivity was comparatively high (0.03gL(-1)d(-1)) due to its high biomass productivity (0.37gL(-1)d(-1)). FAMEs were satisfactory to the several standards for the biodiesel quality. The Scenedesmus-based technology may serve as a promising option for the treatment of nutrient-rich wastewater and especially so for the AD effluent.

Hydrogen fermentation of food waste by alkali-shock pretreatment: microbial community analysis and limitation of continuous operation.

In the study, at first, batch tests were performed to investigate the effect of alkali-shock on H2 production from food waste (FW). After alkali-pretreatment of FW at pH 9.0-13.0, the FW was cultivated under mesophilic condition at pH 6.0 for 30 h without external inoculum addition. The amount of H2 production from FW pretreated at pH 11.0 and 12.0 was higher than that achieved in other pretreatment pH. The main metabolite was butyrate, and Clostridium were dominant at pH 11.0 and 12.0. Meanwhile, lactate was the main metabolite with Enterococcus and Streptococcus being the dominant genus at other pretreatment pH. When the batch process was switched to a continuous mode, H2 production was significantly dropped due to the increased activity of H2-consumers. The reliability of alkali-pretreatment at pH 11.0 was proven by repeating the scale-up batch process, recording 1.57±0.11 mol H2/mol hexose(added) (17±2LH2/kg FW) and 4.39±0.32LH2/L/d.