Impacts of metal stress on extracellular microbial products, and potential for selective metal recovery.

Harnessing microbial capabilities for metal recovery from secondary waste sources is an eco-friendly and sustainable approach for the management of metal-containing wastes. Soluble microbial products (SMP) and extracellular polymeric substances (EPS) are the two main groups of extracellular compounds produced by microorganisms in response to metal stress that are of great importance for remediation and recovery of metals. These include various high-, and low, molecular weight components, which serve various functional and structural roles. These compounds often contain functional groups with metal binding potential that can attenuate metal stress by sequestering metal ions, making them less bioavailable. Microorganisms can regulate the content and composition of EPS and SMP in response to metal stress in order to increase the compounds specificity and capacity for metal binding. Thus, EPS and SMP represent ideal candidates for developing technologies for selective metal recovery from complex wastes. To discover highly metal-sorptive compounds with specific metal binding affinity for metal recovery applications, it is necessary to investigate the metal binding affinity of these compounds, especially under metal stressed conditions. In this review we critically reviewed microbial EPS and SMP production as a response to metal stress with a particular emphasis on the metal binding properties of these compounds and their role in altering metal bioavailability. Furthermore, for the first time, we compiled the available data on potential application of these compounds for selective metal recovery from waste streams.

Dynamic gill and mucus microbiomes during a gill disease episode in farmed Atlantic salmon.

Amoebic gill disease (AGD) and complex gill disease (CGD) are recurrent gill disorders in Atlantic salmon, resulting in significant aquaculture losses. The role of gill microbiomes in gill disease development is unclear. We undertook a longitudinal study to characterise the gill tissue and gill mucus microbiomes of farmed Atlantic salmon before, and during, a gill disease episode. Using a newly optimised DNA extraction protocol, we sequenced rRNA genes from microbiomes of gill samples taken from 105 individual salmon on a farm, over a summer season. The AGD aetiological agent, Neoparamoeba perurans, was PCR-quantified targeting 18S rRNA genes. Similar analyses were carried out on mucus samples. Mucus scrapings were suitable, non-lethal substitutes for characterisation of the gill prokaryotic community in this study. Gill tissue and gill mucus microbiomes changed during the campaign, correlating with N. perurans concentrations. Time explained 35% of the gill tissue and gill mucus microbiome variance, while N. perurans concentrations explained 5%. Genera including Dyadobacter, Shewanella and Pedobacter were maximally abundant in gill and mucus samples at the timepoint prior to the the detection of gill disorder signs, at T3. Shewanella was significantly more abundant before than during the gill disease episode, and we suggest this genus could be considered in future studies addressing relationships between gill disease and the gill microbiome.

A meta-analysis of acetogenic and methanogenic microbiomes in microbial electrosynthesis.

A meta-analysis approach was used, to study the microbiomes of biofilms and planktonic communities underpinning microbial electrosynthesis (MES) cells. High-throughput DNA sequencing of 16S rRNA gene amplicons has been increasingly applied to understand MES systems. In this meta-analysis of 22 studies, we find that acetogenic and methanogenic MES cells share 80% of a cathodic core microbiome, and that different inoculum pre-treatments strongly affect community composition. Oxygen scavengers were more abundant in planktonic communities, and several key organisms were associated with operating parameters and good cell performance. We suggest Desulfovibrio sp. play a role in initiating early biofilm development and shaping microbial communities by catalysing H2 production, to sustain either Acetobacterium sp. or Methanobacterium sp. Microbial community assembly became more stochastic over time, causing diversification of the biofilm (cathodic) community in acetogenic cells and leading to re-establishment of methanogens, despite inoculum pre-treatments. This suggests that repeated interventions may be required to suppress methanogenesis.

Interrelating EPS, soluble microbial products and metal solubility in a methanogenic consortium stressed by nickel and cobalt.

The relationships between extracellular polymeric substances (EPS), soluble microbial product production, metal solubility, and methanogenic activity were investigated. The individual, and joint, toxic effects of nickel and cobalt on methanogenic consortia fed with glucose as model substrate were studied using biomethane potential assays. Cobalt was found to be less toxic to methanogens than nickel at each concentration tested, and the combined effects of Ni and Co on methane production in the bimetal experiment was higher than the sum of the effects of each metal alone. The protein content of EPS, and extracellular soluble protein fractions, decreased with increasing concentrations of total metals. Meanwhile, no significant change in response to metal stress was apparent for carbohydrate content of EPS or extracellular soluble carbohydrate. Decreasing protein content of EPS was accompanied by reduced methanogenic activity and an increase in the soluble metal fraction. The strong associations observed between these variables could be due to the critical role of EPS in protecting microbial cells against nickel and cobalt stress, possibly by capturing metal cations through their functional groups, thus reducing metal availability to the microbial cells in the methanogenic consortia underpinning the anaerobic digestion process.

Microbial community assembly and dynamics in Granular, Fixed-Biofilm and planktonic microbiomes valorizing Long-Chain fatty acids at 20 °C.

Distinct microbial assemblages evolve in anaerobic digestion (AD) reactors to drive sequential conversions of organics to methane. The spatio-temporal development of three such assemblages (granules, biofilms, planktonic) derived from the same inoculum was studied in replicated bioreactors treating long-chain fatty acids (LCFA)-rich wastewater at 20 °C at hydraulic retention times (HRTs) of 12-72 h. We found granular, biofilm and planktonic assemblages differentiated by diversity, structure, and assembly mechanisms; demonstrating a spatial compartmentalisation of the microbiomes from the initial community reservoir. Our analysis linked abundant Methanosaeta and Syntrophaceae-affiliated taxa (Syntrophus and uncultured) to their putative, active roles in syntrophic LCFA bioconversion. LCFA loading rates (stearate, palmitate), and HRT, were significant drivers shaping microbial community dynamics and assembly. This study of the archaea and syntrophic bacteria actively valorising LCFAs at short HRTs and 20 °C will help uncover the microbiology underpinning anaerobic bioconversions of fats, oil and grease.

Biofilm carrier type affects biogenic sulfur-driven denitrification performance and microbial community dynamics in moving-bed biofilm reactors.

Autotrophic denitrification with biosulfur (ADBIOS) provides a sustainable technological solution for biological nitrogen removal from wastewater driven by biogenic S0, derived from biogas desulfurization. In this study, the effect of different biofilm carriers (conventional AnoxK™ 1 and Z-200 with a pre-defined maximum biofilm thickness) on ADBIOS performance and microbiomics was investigated in duplicate moving bed-biofilm reactors (MBBRs). The MBBRs were operated parallelly in continuous mode for 309 days, whilst gradually decreasing the hydraulic retention time (HRT) from 72 to 21 h, and biosulfur was either pumped in suspension (days 92-223) or supplied in powder form. Highest nitrate removal rates were approximately 225 (±11) mg/L·d and 180 (±7) mg NO3--N/L·d in the MBBRs operated with K1 and Z-200 carriers, respectively. Despite having the same protected surface area for biofilm development in each MBBR, the biomass attached onto the K1 carrier was 4.8-fold more than that on the Z-200 carrier, with part of the biogenic S0 kept in the biofilm. The microbial communities of K1 and Z-200 biofilms could also be considered similar at cDNA level in terms of abundance (R = 0.953 with p = 0.042). A relatively stable microbial community was formed on K1 carriers, while the active portion of the microbial community varied significantly over time in the MBBRs using Z-200 carriers.

A Distinct, Flocculent, Acidogenic Microbial Community Accompanies Methanogenic Granules in Anaerobic Digesters.

The formation of dense, well-settling methanogenic granules is essential for the operation of high-rate, up-flow anaerobic bioreactors used for wastewater treatment. Granule formation (granulation) mechanisms have been previously proposed, but an ecological understanding of granule formation is still lacking. Additionally, much of the current research on granulation only examines the start-up phase of bioreactor operation, rather than monitoring the fate of established granules and how new granules emerge over time. This paper, therefore, attempts to provide an insight into the microbial ecology of granule formation outside the start-up phase of bioreactor operation and develop an ecological granulation model. The microbial communities of granules actively undergoing growth, breakage, and reformation were examined, and an ecological granulation model was proposed. A distinct pregranular microbial community, with a high proportion of acidogenic organisms, such as the Streptococcaceae, was identified and suggested to have a role in initiating granulation by providing simpler substrates for the methanogenic and syntrophic communities which developed during granule growth. After initial granule formation, deterministic influences on microbial community assembly increased with granule size and indicated that microbial community succession was influenced by granule growth, leading to the formation of a stepwise ecological model for granulation. IMPORTANCE Complex microbial communities in engineered environments can aggregate to form surface-attached biofilms. Others form suspended biofilms, such as methanogenic granules. The formation of dense, methanogenic granules underpins the performance of high-rate, anaerobic bioreactors in industrial wastewater treatment. Granule formation (granulation) has been well studied from a physico-chemical perspective, but the ecological basis is poorly understood. We identified a distinct, flocculent, microbial community, which was present alongside granules, comprising primary consumers likely key in providing simpler substrates to granules. This flocculent community is understudied in anaerobic digestion and may initiate, or perpetuate, granule formation. We propose that it may be possible to influence bioreactor performance (e.g., to regulate volatile fatty acid concentrations) by manipulating this community. The patterns of microbial community diversity and assembly revealed by the study indicate that cycles of granule growth and breakage lead to overall diversification of the bioreactor meta-community, with implications for bioreactor process stability.

Bayesian Generalized Linear Mixed-Model Analysis of Language Samples: Detecting Patterns in Expository and Narrative Discourse of Adolescents With Traumatic Brain Injury.

Purpose Generalized linear mixed-model (GLMM) and Bayesian methods together provide a framework capable of handling a wide variety of complex data commonly encountered across the communication sciences. Using language sample analysis, we demonstrate the utility of these methods in answering specific questions regarding the differences between discourse patterns of children who have experienced a traumatic brain injury (TBI), as compared to those with typical development. Method Language samples were collected from 55 adolescents ages 13-18 years, five of whom had experienced a TBI. We describe parameters relating to the productivity, syntactic complexity, and lexical diversity of language samples. A Bayesian GLMM is developed for each parameter of interest, relating these parameters to age, sex, prior history (TBI or typical development), and socioeconomic status, as well as the type of discourse sample (compare-contrast, cause-effect, or narrative). Statistical models are thoroughly described. Results Comparing the discourse of adolescents with TBI to those with typical development, substantial differences are detected in productivity and lexical diversity, while differences in syntactic complexity are more moderate. Female adolescents exhibited greater syntactic complexity, while male adolescents exhibited greater productivity and lexical diversity. Generally, our models suggest more advanced discourse among adolescents who are older or who have indicators of higher socioeconomic status. Differences relating to lecture type were also detected. Conclusions Bayesian and GLMM methods yield more informative and intuitive results than traditional statistical analyses, with a greater degree of confidence in model assumptions. We recommend that these methods be used more widely in language sample analysis. Supplemental Material https://doi.org/10.23641/asha.14226959.

Methanogenic granule growth and development is a continual process characterized by distinct morphological features.

Up-flow anaerobic bioreactors are widely applied for high-rate digestion of industrial wastewaters and rely on formation, and retention, of methanogenic granules, comprising of dense, fast-settling, microbial aggregates (approx. 0.5-4.0 mm in diameter). Granule formation (granulation) mechanisms have been reasonably well hypothesized and documented. However, this study used laboratory-scale bioreactors, inoculated with size-separated granular sludge to follow new granule formation, maturation, disintegration and re-formation. Temporal size profiles, volatile solids content, settling velocity, and ultrastructure of granules were determined from each of four bioreactors inoculated only with small granules, four with only large granules, and four with a full complement of naturally-size-distributed granules. Constrained granule size profiles shifted toward the natural distribution, which was associated with maximal bioreactor performance. Distinct morphological features characterized different granule sizes and biofilm development stages, including 'young', 'juvenile', 'mature' and 'old'. The findings offer opportunities toward optimizing management of high-rate, anaerobic digesters by shedding light on the rates of granule growth, the role of flocculent sludge in granulation and how shifting size distributions should be considered when setting upflow velocities.

Carboxylic acids production and electrosynthetic microbial community evolution under different CO2 feeding regimens.

Microbial electrosynthesis (MES) is a potential technology for CO2 recycling, but insufficient information is available on the microbial interactions underpinning electrochemically-assisted reactions. In this study, a MES reactor was operated for 225 days alternately with bicarbonate or CO2 as carbon source, under batch or continuous feeding regimens, to evaluate the response of the microbial communities, and their productivity, to dynamic operating conditions. A stable acetic acid production rate of 9.68 g m-2 d-1, and coulombic efficiency up to 40%, was achieved with continuous CO2 sparging, higher than the rates obtained with bicarbonate (0.94 g m-2 d-1) and CO2 under fed-batch conditions (2.54 g m-2 d-1). However, the highest butyric acid production rate (0.39 g m-2 d-1) was achieved with intermittent CO2 sparging. The microbial community analyses focused on differential amplicon sequence variants (ASVs), allowing detection of ASVs significantly different across consecutive samples. This analysis, combined with co-occurence network analysis, and cyclic voltammetry, indicated that hydrogen-mediated acetogenesis was carried out by Clostridium, Eubacterium and Acetobacterium, whereas Oscillibacter and Caproiciproducens were involved in butyric acid production. The cathodic community was spatially inhomogeneous, with potential electrotrophs, such as Sulfurospirillum and Desulfovibrio, most prevalent near the current collector. The abundance of Sulfurospirillum positively correlated with that of Acetobacterium, supporting the syntrophic metabolism of both organisms.

Size Shapes the Active Microbiome of Methanogenic Granules, Corroborating a Biofilm Life Cycle.

Methanogenic archaea are key players in cycling organic matter in nature but also in engineered waste treatment systems, where they generate methane, which can be used as a renewable energy source. In such systems in the built environment, complex methanogenic consortia are known to aggregate into highly organized, spherical granular biofilms comprising the interdependent microbial trophic groups mediating the successive stages of the anaerobic digestion (AD) process. This study separated methanogenic granules into a range of discrete size fractions, hypothesizing different biofilm growth stages, and separately supplied each with specific substrates to stimulate the activity of key AD trophic groups, including syntrophic acid oxidizers and methanogens. Rates of specific methanogenic activity were measured, and amplicon sequencing of 16S rRNA gene transcripts was used to resolve phylotranscriptomes across the series of size fractions. Increased rates of methane production were observed in each of the size fractions when hydrogen was supplied as the substrate compared with those of volatile fatty acids (acetate, propionate, and butyrate). This was connected to a shift toward hydrogenotrophic methanogenesis dominated by Methanobacterium and Methanolinea Interestingly, the specific active microbiomes measured in this way indicated that size was significantly more important than substrate in driving the structure of the active community in granules. Multivariate integration studywise discriminant analysis identified 56 genera shaping changes in the active community across both substrate and size. Half of those were found to be upregulated in the medium-sized granules, which were also the most active and potentially of the most important size, or life stage, for precision management of AD systems.IMPORTANCE Biological wastewater conversion processes collectively constitute one of the single biggest worldwide applications of microbial communities. There is an obvious requirement, therefore, to study the microbial systems central to the success of such technologies. Methanogenic granules, in particular, are architecturally fascinating biofilms that facilitate highly organized cooperation within the metabolic network of the anaerobic digestion (AD) process and, thus, are especially intriguing model systems for microbial ecology. This study, in a way not previously reported, provoked syntrophic and methanogenic activity and the structure of the microbial community, using specific substrates targeting the key trophic groups in AD. Unexpectedly, granule size more strongly than substrate shaped the active portion of the microbial community. Importantly, the findings suggest the size, or age, of granules inherently shapes the active microbiome linked to a life cycle. This provides exciting insights into the function of, and the potential for additional modeling of biofilm development in, methanogenic granules.

Growth and Break-Up of Methanogenic Granules Suggests Mechanisms for Biofilm and Community Development.

Methanogenic sludge granules are densely packed, small, spherical biofilms found in anaerobic digesters used to treat industrial wastewaters, where they underpin efficient organic waste conversion and biogas production. Each granule theoretically houses representative microorganisms from all of the trophic groups implicated in the successive and interdependent reactions of the anaerobic digestion (AD) process. Information on exactly how methanogenic granules develop, and their eventual fate will be important for precision management of environmental biotechnologies. Granules from a full-scale bioreactor were size-separated into small (0.6-1 mm), medium (1-1.4 mm), and large (1.4-1.8 mm) size fractions. Twelve laboratory-scale bioreactors were operated using either small, medium, or large granules, or unfractionated sludge. After >50 days of operation, the granule size distribution in each of the small, medium, and large bioreactor sets had diversified beyond-to both bigger and smaller than-the size fraction used for inoculation. Interestingly, extra-small (XS; <0.6 mm) granules were observed, and retained in all of the bioreactors, suggesting the continuous nature of granulation, and/or the breakage of larger granules into XS bits. Moreover, evidence suggested that even granules with small diameters could break. "New" granules from each emerging size were analyzed by studying community structure based on high-throughput 16S rRNA gene sequencing. Methanobacterium, Aminobacterium, Propionibacteriaceae, and Desulfovibrio represented the majority of the community in new granules. H2-using, and not acetoclastic, methanogens appeared more important, and were associated with abundant syntrophic bacteria. Multivariate integration (MINT) analyses identified distinct discriminant taxa responsible for shaping the microbial communities in different-sized granules.

Predicting vertical ground reaction force during running using novel piezoresponsive sensors and accelerometry.

Running is a common exercise with numerous health benefits. Vertical ground reaction force (vGRF) influences running injury risk and running performance. Measurement of vGRF during running is now primarily constrained to a laboratory setting. The purpose of this study was to evaluate a new approach to measuring vGRF during running. This approach can be used outside of the laboratory and involves running shoes instrumented with novel piezoresponsive sensors and a standard accelerometer. Thirty-one individuals ran at three different speeds on a force-instrumented treadmill while wearing the instrumented running shoes. vGRF was predicted using data collected from the instrumented shoes, and predicted vGRF were compared to vGRF measured via the treadmill. Per cent error of the resulting predictions varied depending upon the predicted vGRF characteristic. Per cent error was relatively low for predicted vGRF impulse (2-7%), active peak vGRF (3-7%), and ground contact time (3-6%), but relatively high for predicted vGRF load rates (22-29%). These errors should decrease with future iterations of the instrumented shoes and collection of additional data from a more diverse sample. The novel technology described herein might become a feasible way to collect large amounts of vGRF data outside of the traditional biomechanics laboratory.

Propionate Production by Bioelectrochemically-Assisted Lactate Fermentation and Simultaneous CO2 Recycling.

Production of volatile fatty acids (VFAs), fundamental building blocks for the chemical industry, depends on fossil fuels but organic waste is an emerging alternative substrate. Lactate produced from sugar-containing waste streams can be further processed to VFAs. In this study, electrofermentation (EF) in a two-chamber cell is proposed to enhance propionate production via lactate fermentation. At an initial pH of 5, an applied potential of -1 V vs. Ag/AgCl favored propionate production over butyrate from 20 mM lactate (with respect to non-electrochemical control incubations), due to the pH buffering effect of the cathode electrode, with production rates up to 5.9 mM d-1 (0.44 g L-1 d-1). Microbial community analysis confirmed the enrichment of propionate-producing microorganisms, such as Tyzzerella sp. and Propionibacterium sp. Organisms commonly found in microbial electrosynthesis reactors, such as Desulfovibrio sp. and Acetobacterium sp., were also abundant at the cathode, indicating their involvement in recycling CO2 produced by lactate fermentation into acetate, as confirmed by stoichiometric calculations. Propionate was the main product of lactate fermentation at substrate concentrations up to 150 mM, with a highest production rate of 12.9 mM d-1 (0.96 g L-1 d-1) and a yield of 0.48 mol mol-1 lactate consumed. Furthermore, as high as 81% of the lactate consumed (in terms of carbon) was recovered as soluble product, highlighting the potential for EF application with high-carbon waste streams, such as cheese whey or other food wastes. In summary, EF can be applied to control lactate fermentation toward propionate production and to recycle the resulting CO2 into acetate, increasing the VFA yield and avoiding carbon emissions and addition of chemicals for pH control.

Physically constrained spatiotemporal modeling: generating clear-sky constructions of land surface temperature from sparse, remotely sensed satellite data.

Satellite remote-sensing is used to collect important atmospheric and geophysical data at various spatial resolutions, providing insight into spatiotemporal surface and climate variability globally. These observations are often plagued with missing spatial and temporal information of Earth's surface due to (1) cloud cover at the time of a satellite passing and (2) infrequent passing of polar-orbiting satellites. While many methods are available to model missing data in space and time, in the case of land surface temperature (LST) from thermal infrared remote sensing, these approaches generally ignore the temporal pattern called the 'diurnal cycle' which physically constrains temperatures to peak in the early afternoon and reach a minimum at sunrise. In order to infill an LST dataset, we parameterize the diurnal cycle into a functional form with unknown spatiotemporal parameters. Using multiresolution spatial basis functions, we estimate these parameters from sparse satellite observations to reconstruct an LST field with continuous spatial and temporal distributions. These estimations may then be used to better inform scientists of spatiotemporal thermal patterns over relatively complex domains. The methodology is demonstrated using data collected by MODIS on NASA's Aqua and Terra satellites over both Houston, TX and Phoenix, AZ USA.

Microbial diversity and long-term geochemical trends in the euxinic zone of a marine, meromictic lake.

Hypoxic and anoxic niches of meromictic lakes are important sites for studying the microbial ecology of conditions resembling ancient Earth. The expansion and increasing global distribution of such environments also means that information about them serves to understand future phenomena. In this study, a long-term chemical dataset (1996-2015) was explored together with seasonal (in 2015) information on the diversity and abundance of bacterial and archaeal communities residing in the chemocline, monimolimnion and surface sediment of the marine meromictic Rogoznica Lake. The results of quantitative PCR assays, and high-throughput sequencing, targeting 16S rRNA genes and transcripts, revealed a clear vertical structure of the microbial community with Gammaproteobacteria (Halochromatium) and cyanobacteria (Synechococcus spp.) dominating the chemocline, Deltaproteobacteria and Bacteroidetes dominating the monimolimnion, and significantly more abundant archaeal populations in the surface sediment, most of which affiliated to Nanoarchaeota. Seasonal changes in the community structure and abundance were not pronounced. Diversity in Rogoznica Lake was found to be high, presumably as a consequence of stable environmental conditions accompanied by high dissolved carbon and nutrient concentrations. Long-term data indicated that Rogoznica Lake exhibited climate changes that could alter its physico-chemical features and, consequently, induce structural and physiological changes within its microbial community.

Power production and microbial community composition in thermophilic acetate-fed up-flow and flow-through microbial fuel cells.

The microbial communities developed from a mixed-species culture in up-flow and flow-through configurations of thermophilic (55 °C) microbial fuel cells (MFCs), and their power production from acetate, were investigated. The up-flow MFC was operated for 202 days, obtaining an average power density of 0.13 W/m3, and Tepidiphilus was the dominant transcriptionally-active microorganisms. The planktonic community developed in the up-flow MFC was used to inoculate a flow-through MFC resulting in the proliferation of Ureibacillus, whose relative abundance increased from 1 to 61% after 45 days. Despite the differences between the up-flow and flow-through MFCs, including the anode electrode, hydrodynamic conditions, and the predominant microorganism, similar (p = 0.05) volumetric power (0.11-0.13 W/m3), coulombic efficiency (16-18%) and acetate consumption rates (55-69 mg/L/d) were obtained from both. This suggests that though MFC design can shape the active component of the thermophilic microbial community, the consortia are resilient and can maintain similar performance in different MFC configurations.

Anaerobic treatment of LCFA-containing synthetic dairy wastewater at 20 °C: Process performance and microbial community dynamics.

Facilitating anaerobic degradation of long-chain fatty acids (LCFA) is key for tapping the high methane production potential of the fats, oil and grease (FOG) content of dairy wastewaters. In this study, the feasibility of using high-rate granular sludge reactors for the treatment of mixed LCFA-containing synthetic dairy wastewater (SDW) was assessed at 20 °C. The effects of the LCFA concentration (33-45% of COD) and organic loading rates (2-3 gCOD/L·d) were determined using three parallel expanded granular sludge bed reactors. For the first time, long term anaerobic treatment of LCFA-containing feed at 20 °C was shown to be feasible and was linked to the microbial community dynamics in high-rate reactors. During a two-month operation, a soluble COD removal of 84-91% and COD to methane conversion of 44-51% was obtained. However, granular sludge flotation and washout occurred after two months in all reactors without volatile fatty acids (VFA) accumulation, emphasizing the need for sludge retention for long-term granular sludge reactor operation with LCFA-containing feed at low ambient temperatures. The temporal shifts in microbial community structure were studied in the high-rate treatment of SDW, and the process disturbances (elevated LCFA loading, LCFA accumulation, and batch operation) were found to decrease the microbial community diversity. The relative abundance of Methanosaeta increased with higher LCFA accumulation in the settled and flotation layer granules in the three reactors, therefore, acetoclastic methanogenesis was found to be crucial for the high-rate treatment of SDW at 20 °C. This study provides an initial understanding of the continuous anaerobic treatment of LCFA-containing industrial wastewaters at low ambient temperatures.

Acetotrophic Activity Facilitates Methanogenesis from LCFA at Low Temperatures: Screening from Mesophilic Inocula.

The inoculum source plays a crucial role in the anaerobic treatment of wastewaters. Lipids are present in various wastewaters and have a high methanogenic potential, but their hydrolysis results in the production of long chain fatty acids (LCFAs) that are inhibitory to anaerobic microorganisms. Screening of inoculum for the anaerobic treatment of LCFA-containing wastewaters has been performed at mesophilic and thermophilic conditions. However, an evaluation of inocula for producing methane from LCFA-containing wastewater has not yet been conducted at low temperatures and needs to be undertaken. In this study, three inocula (one granular sludge and two municipal digester sludges) were assessed for methane production from LCFA-containing synthetic dairy wastewater (SDW) at low temperatures (10 and 20°C). A methane yield (based on mL-CH4/g-CODadded) of 86-65% with acetate and 45-20% with SDW was achieved within 10 days using unacclimated granular sludge, whereas the municipal digester sludges produced methane only at 20°C but not at 10°C even after 200 days of incubation. The acetotrophic activity in the inoculum was found to be crucial for methane production from LCFA at low temperatures, highlighting the role of Methanosaeta (acetoclastic archaea) at low temperatures. The presence of bacterial taxa from the family Syntrophaceae (Syntrophus and uncultured taxa) in the inoculum was found to be important for methane production from SDW at 10°C. This study suggests the evaluation of acetotrophic activity and the initial microbial community characteristics by high-throughput amplicon sequencing for selecting the inoculum for producing methane at low temperatures (up to 10°C) from lipid-containing wastewaters.