Rheological properties of thermally treated and digested sludge: Implications for energy requirements of pumps and agitators.

Understanding sludge rheology and optimizing equipment performance is crucial for energy efficiency in wastewater treatment plants (WWTPs). This study examined sludge rheology after thermal hydrolysis pre-treatment (THP) at 60, 80, and 120 °C for 2 h, followed by anaerobic digestion (AD) at 37 °C for 20 days, and assessed impacts on pump and agitator performance. Post-treatment, sludge showed reduced viscosity and improved flowability, indicated by changes in Herschel-Bulkley parameters, enhancing pump and agitator efficiency, particularly at 120 °C. These rheological improvements were correlated to the solubilization of sludge components after THP and solids reduction after AD, highlighting the interconnectedness of rheology and treatment outcomes. Despite high heat demands, an energy balance showed that THP scenarios, especially at 120 °C, had lower energy requirements for pumps and agitators, leading to energy savings without increased heat consumption. These findings underscore the influence of rheological changes in improving energy efficiency in WWTPs.

ALACEN: A Holistic Herbaceous Biomass Fractionation Process Attaining a Xylose-Rich Stream for Direct Microbial Conversion to Bioplastics.

Lignocellulose biorefining is a promising technology for the sustainable production of chemicals and biopolymers. Usually, when one component is focused on, the chemical nature and yield of the others are compromised. Thus, one of the bottlenecks in biomass biorefining is harnessing the maximum value from all of the lignocellulosic components. Here, we describe a mild stepwise process in a flow-through setup leading to separate flow-out streams containing cinnamic acid derivatives, glucose, xylose, and lignin as the main components from different herbaceous sources. The proposed process shows that minimal degradation of the individual components and conservation of their natural structure are possible. Under optimized conditions, the following fractions are produced from wheat straw based on their respective contents in the feed by the ALkaline ACid ENzyme process: (i) 78% ferulic acid from a mild ALkali step, (ii) 51% monomeric xylose free of fermentation inhibitors by mild ACidic treatment, (iii) 82% glucose from ENzymatic degradation of cellulose, and (iv) 55% native-like lignin. The benefits of using the flow-through setup are demonstrated. The retention of the lignin aryl ether structure was confirmed by HSQC NMR, and this allowed monomers to form from hydrogenolysis. More importantly, the crude xylose-rich fraction was shown to be suitable for producing polyhydroxybutyrate bioplastics. The direct use of the xylose-rich fraction by means of the thermophilic bacteria Schlegelella thermodepolymerans matched 91% of the PHA produced with commercial pure xylose, achieving 138.6 mgPHA/gxylose. Overall, the ALACEN fractionation method allows for a holistic valorization of the principal components of herbaceous biomasses.

Polyhydroxyalkanoates (PHAs) synthesis and degradation by microbes and applications towards a circular economy.

Overusing non-degradable plastics causes a series of environmental issues, inferring a switch to biodegradable plastics. Polyhydroxyalkanoates (PHAs) are promising biodegradable plastics that can be produced by many microbes using various substrates from waste feedstock. However, the cost of PHAs production is higher compared to fossil-based plastics, impeding further industrial production and applications. To provide a guideline for reducing costs, the potential cheap waste feedstock for PHAs production have been summarized in this work. Besides, to increase the competitiveness of PHAs in the mainstream plastics economy, the influencing parameters of PHAs production have been discussed. The PHAs degradation has been reviewed related to the type of bacteria, their metabolic pathways/enzymes, and environmental conditions. Finally, the applications of PHAs in different fields have been presented and discussed to induce comprehension on the practical potentials of PHAs.

The impact of carbon to nitrogen ratios and pH on the microbial prevalence and polyhydroxybutyrate production levels using a mixed microbial starter culture.

Growth conditions have been frequently studied in optimizing polyhydroxybutyrate (PHB) production, while few studies were performed to unravel the dynamic mixed microbial consortia (MMCs) in the process. In this study, the relationship between growth conditions (C/N ratios and pH) and the corresponding key-microbes were identified and monitored during PHB accumulation. The highest PHB level (70 wt% of dry cell mass) was obtained at pH 9, C/N 40, and acetic acid 10 g/L. Linking the dominant genera with the highest point of PHB accumulation, Thauera was the most prevalent species in all MMCs of pH 9, except when a C/N ratio of 1 was applied. Notably, dominant bacteria shifted at pH 7 (C/N 10) from Thauera (0 h) to Paracoccus, and subsequently to Alcaligenes following the process of PHB accumulation and consumption. Further understanding of the relationship between the structure of the microbial community and the performance will be beneficial for regulating and obtaining high PHB accumulation within an MMC. Our study illustrates the impact of C/N ratios and pH on microbial prevalence and PHB production levels using a mixed microbial starter culture. This knowledge will broaden industrial perspectives for regulating high PHB production and timely harvesting.

Strategies to boost anaerobic digestion performance of cow manure: Laboratory achievements and their full-scale application potential.

Cow manure represents a surplus manure waste in agricultural food sectors, which requires proper disposal. Anaerobic digestion, in this regard, has raised global interest owing to its apparent environmental benefits, including simultaneous waste diminishment and renewable energy generation. However, dedicated intensifications are necessary to promote the degradation of recalcitrant lignocellulosic components of cow manure. Hence, this manuscript presents a review of how to exploit cow manure in anaerobic digestion through different incentives extensively at lab-scale and full-scale. These strategies comprise 1) co-digestion; 2) pretreatment; 3) introduction of additives (trace metals, carbon-based materials, low-cost composites, nanomaterials, and microbial cultures); 4) innovative systems (bio-electrochemical fields and laser irradiation). Results imply that co-digestion and pretreatment approaches gain the predominance on promoting the digestion performance of cow manure. Particularly, for the co-digestion scenario, the selection of lignin-poor co-substrate is highlighted to produce maximum synergy and pronounced removal of lignocellulosic compounds of cow manure. Mechanical, thermal, and biological (composting) pretreatments generate mild improvement at laboratory-scale and are proved applicable in full-scale facilities. It is noteworthy that the introduction of additives (Fe-based nanomaterials, carbon-based materials, and composites) is acquiring more attention and shows promising full-scale application potential. Finally, bio-electrochemical fields stand out in laboratory trials and may serve as future reactor modules in agricultural anaerobic digestion installations treating cow manure.

Comparison of the microbial communities in anaerobic digesters treating high alkalinity synthetic wastewater at atmospheric and high-pressure (11 bar).

High-pressure anaerobic digestion is an appealing concept since it can upgrade biogas directly within the reactor. However, the decline of pH caused by the dissolution of CO2 is the main barrier that prevents a good operating high-pressure anaerobic digestion process. Therefore, in this study, a high-pressure anaerobic digestion was studied to treat high alkalinity synthetic wastewater, which could not be treated in a normal-pressure anaerobic digester. In the high-pressure reactor, the pH value was 7.5 ~ 7.8, and the CH4 content reached 88% at 11 bar. Unlike its normal-pressure counterpart (2285 mg/L acetic acid), the high-pressure reactor ran steadily (without volatile fatty acids inhibition). Furthermore, the microbial community changed in the high-pressure reactor. Specifically, key microbial guilds (Syntrophus (11.2%), Methanosaeta concilii (50.9%), and Methanobrevibacter (26.8%)) were dominant in the high-pressure reactor at 11 bar, indicating their fundamental roles under high-pressure treating high alkalinity synthetic wastewater.

Miniaturization and 3D Printing of Bioreactors: A Technological Mini Review.

Many articles have been published on scale-down concepts as well as additive manufacturing techniques. However, information is scarce when miniaturization and 3D printing are applied in the fabrication of bioreactor systems. Therefore, garnering information for the interfaces between miniaturization and 3D printing becomes important and essential. The first goal is to examine the miniaturization aspects concerning bioreactor screening systems. The second goal is to review successful modalities of 3D printing and its applications in bioreactor manufacturing. This paper intends to provide information on anaerobic digestion process intensification by fusion of miniaturization technique and 3D printing technology. In particular, it gives a perspective on the challenges of 3D printing and the options of miniature bioreactor systems for process high-throughput screening.

The biomethanation of cow manure in a continuous anaerobic digester can be boosted via a bioaugmentation culture containing Bathyarchaeota.

A bioaugmentation approach was used to enhance the performance of anaerobic digestion (AD) using cow manure (CM) as the substrate in a continuous system. To obtain the desirable microbial culture for bioaugmentation, a biochemical methane potential test (BMP) was used to evaluate three commonly used inocula namely (1) municipal solid waste (MSW), (2) wastewater treatment plant (WWTP), and (3) cow manure digester (CMMD) for their hydrolytic capacity. The highest lignocellulose removal (56% for cellulose and 50% for hemicellulose) and the most profusion of cellulolytic bacteria were obtained when CM was inoculated with CMMD. CMMD was thus used as the seed inoculum in a continuously operated reactor (Ra) with the fiber fraction of CM as the substrate to further enrich cellulolytic microbes. After 100 days (HRT: 30 days), the Bacteria fraction mainly contained Ruminofilibacter, norank_o_SBR1031, Treponema, Acetivibrio. Surprisingly, the Archaea fraction contained 97% 'cellulolytic archaea' norank_c_Bathyarchaeia (Phylum Bathyarchaeota). This enriched consortium was used in the bioaugmentation experiment. A positive effect of bioaugmentation was verified, with a substantial daily methane yield (DMY) enhancement (24.3%) obtained in the bioaugmented reactor (Rb) (179 mL CH4/gVS/d) than that of the control reactor (Rc) (144 mL CH4/gVS/d) (P < 0.05). Meanwhile, the effluent of Rb enjoyed an improved cellulose reduction (14.7%) than that of Rc, whereas the amount of hemicellulose remained similar in both reactors' effluent. When bioaugmentation stopped, its influence on the hydrolysis and methanogenesis sustained, reflected by an improved DMY (160 mL CH4/gVS/d) and lower cellulose content (53 mg/g TS) in Rb than those in Rc (DMY 144 mL/CH4/gVS/d and cellulose content 63 mg/g TS, respectively). The increased DMY of the continuous reactor seeded with a specifically enriched consortium able to degrade the fiber fraction in CM shows the feasibility of applying bioaugmentation in AD of CM.

A Technological Understanding of Biofilm Detection Techniques: A Review.

Biofouling is a persistent problem in almost any water-based application in several industries. To eradicate biofouling-related problems in bioreactors, the detection of biofilms is necessary. The current literature does not provide clear supportive information on selecting biofilm detection techniques that can be applied to detect biofouling within bioreactors. Therefore, this research aims to review all available biofilm detection techniques and analyze their characteristic properties to provide a comparative assessment that researchers can use to find a suitable biofilm detection technique to investigate their biofilms. In addition, it discusses the confluence of common bioreactor fabrication materials in biofilm formation.

Thauera aminoaromatica MZ1T Identified as a Polyhydroxyalkanoate-Producing Bacterium within a Mixed Microbial Consortium.

Polyhydroxyalkanoates (PHAs) form a highly promising class of bioplastics for the transition from fossil fuel-based plastics to bio-renewable and biodegradable plastics. Mixed microbial consortia (MMC) are known to be able to produce PHAs from organic waste streams. Knowledge of key-microbes and their characteristics in PHA-producing consortia is necessary for further process optimization and direction towards synthesis of specific types of PHAs. In this study, a PHA-producing mixed microbial consortium (MMC) from an industrial pilot plant was characterized and further enriched on acetate in a laboratory-scale selector with a working volume of 5 L, and 16S-rDNA microbiological population analysis of both the industrial pilot plant and the 5 L selector revealed that the most dominant species within the population is Thauera aminoaromatica MZ1T, a Gram-negative beta-proteobacterium belonging to the order of the Rhodocyclales. The relative abundance of this Thauera species increased from 24 to 40% after two months of enrichment in the selector-system, indicating a competitive advantage, possibly due to the storage of a reserve material such as PHA. First experiments with T. aminoaromatica MZ1T showed multiple intracellular granules when grown in pure culture on a growth medium with a C:N ratio of 10:1 and acetate as a carbon source. Nuclear magnetic resonance (NMR) analyses upon extraction of PHA from the pure culture confirmed polyhydroxybutyrate production by T. aminoaromatica MZ1T.

Thermal design and turbidity sensor for autonomous bacterial growth measurements in spaceflight.

For application of biological air filters in manned spacecraft, research on bacterial growth is carried out under microgravity conditions. For the BIOFILTER experiment, flown in 2005 on FOTON M2, eight turbidity sensors to measure the growth rate of the bacterium Xanthobacter autotrophicus GJ10 were used. Also thermal management provisions were implemented to control the internal temperature. The design and performance of the BIOFILTER equipment as well as results of the biological ground reference experiments performed in 2006 are discussed. High-performance thermal (vacuum) insulation (lambda= 0.7 mW/mK) and phase change material were implemented, keeping the BIOFILTER internal temperature below 16 degrees C during the 4-day integration period between transport and launch. After launch, in microgravity, the growth of X. autotrophicus GJ10 was successfully triggered by a temperature increase by using an internal heater to 26 degrees C. Although the operation of the sensor electronics was not fully satisfying, the bacterial growth was measured with the sensors, revealing growth rates between 0.046 and 0.077 h(-1) in microgravity, that is, approximately 1.5-2.5 times slower than routinely measured on Earth under optimal laboratory conditions. For the ground-reference experiments the equipment box, containing the eight sensors, was placed on a random positioning machine performing random rotations at 0.5 degrees /min (settling compensation) and 90 degrees /min (microgravity simulation) while the environment was controlled, accurately repeating the BIOFILTER internal temperature profile. Despite the rotation speed differences, growth rates of 0.115 h(-1) were confirmed by both the ground reference experiments. Biological interpretation of the measurements is, however, compromised owing to poor mixing and other unknown physical and biological phenomena that need to be addressed for further space experiments using these kinds of systems.

Identity, abundance and ecophysiology of filamentous bacteria belonging to the Bacteroidetes present in activated sludge plants.

Filamentous members of the Bacteroidetes are commonly observed in activated sludge samples originating from both municipal and industrial wastewater treatment plants (WWTP), where they occasionally can cause bulking. Several oligonucleotide 16S rRNA-targeted probes were designed to target filaments with a needle-like appearance similar to Haliscomenobacter hydrossis. The design of these probes was based on an isolate and a sequence obtained from a micromanipulated filament. The abundance of filamentous Bacteroidetes was determined in 126 industrial samples applying already published and the newly developed probes. Small populations were found in 62 % of the WWTP investigated. However, only relatively few WWTP (13 %) contained large populations of filamentous Bacteroidetes potentially responsible for bulking incidences. The identity of the most abundant filamentous Bacteroidetes with H. hydrossis morphology could be detected by probes CFB719, SAP-309 and the newly designed probe HHY-654. A comprehensive study on the ecophysiology of probe-defined Bacteroidetes populations was conducted on Danish and Czech samples. The studies revealed that they were specialized bacteria involved in degradation of sugars, e.g. glucose and N-acetylglucosamine, and may participate in the conversion of lipopolysaccharides and peptidoglycan liberated by decaying cells. Many surface-associated exo-enzymes were excreted, e.g. chitinase, glucuronidase, esterase and phosphatase, supporting conversion of polysaccharides and possibly other released cell components. The role of filamentous bacteria with a H. hydrossis-like morphology in the activated sludge ecosystem is discussed.

Identity, abundance and ecophysiology of filamentous Chloroflexi species present in activated sludge treatment plants.

Filamentous Chloroflexi species are often present in activated sludge wastewater treatment plants in relatively low numbers, although bulking incidences caused by Chloroflexi filaments have been observed. A new species-specific gene probe for FISH was designed and using phylum-, subdivision-, morphotype 1851- and species-specific gene probes, the abundance of Chloroflexi filaments were monitored in samples from 126 industrial wastewater treatment plants from five European countries. Chloroflexi filaments were present in 50% of the samples, although in low quantities. In most treatment plants the filaments could only be identified with phylum or subdivision probes, indicating the presence of great undescribed biodiversity. The ecophysiology of various Chloroflexi filaments was investigated by a suite of in situ methods. The experiments revealed that Chloroflexi constituted a specialized group of filamentous bacteria only active under aerobic conditions consuming primarily carbohydrates. Many exo-enzymes were excreted, e.g. chitinase, glucuronidase and galactosidase, suggesting growth on complex polysaccharides. The surface of Chloroflexi filaments appeared to be hydrophilic compared to other filaments present. These results are generally supported by physiological studies of two new isolates. Based on the results obtained in this study, the potential role of filamentous Chloroflexi species in activated sludge is discussed.

Phylogeny, physiology and distribution of 'Candidatus Microthrix calida', a new Microthrix species isolated from industrial activated sludge wastewater treatment plants.

Twelve strains of filamentous bacteria morphologically identified as 'Microthrix parvicella' were isolated from industrial activated sludge wastewater treatment plants. 16S rRNA gene sequences analysis showed that these strains were all closely related to 'Candidatus Microthrix parvicella'. Six of them, however, had a 16S rRNA gene similarity of only 95.7% and 96.7% to 'Candidatus Microthrix parvicella' suggesting the presence of a new species. The name 'Candidatus Microthrix calida' is proposed for this new microorganism. The physiological properties of these six isolates supported the description of a new taxon. The 'Candidatus Microthrix calida' strains produced thin filaments (0.3-0.7 microm diameter), they did not grow on the media supporting the growth of 'Candidatus Microthrix parvicella' and could be cultivated at higher temperature (up to 36.5 degrees C). Preliminary data on substrate uptake were obtained by microautoradiography on pure culture. Two new fluorescence in situ hybridization (FISH) probes, Mpa-T1-1260 specific for 'Candidatus Microthrix calida' and Mpa-all-1410 targeting both Microthrix species, were designed. The presence of Microthrix spp. was investigated in 114 activated sludge plants. 'Microthrix parvicella' morphotype was detected in 23% of the analysed samples and FISH analysis revealed that 'Candidatus Microthrix calida' was present in 5% of them. The remaining 'M. parvicella' filaments were positive with probe Mpa-all-1410 but could not all be identified as 'Candidatus Microthrix parvicella' suggesting the presence of more hitherto undescribed biodiversity within this morphotype.

Biofilm formation by Escherichia coli is stimulated by synergistic interactions and co-adhesion mechanisms with adherence-proficient bacteria.

Laboratory strains of Escherichia coli do not show significant ability to attach to solid surfaces and to form biofilms. We compared the adhesion properties of the E. coli PHL565 laboratory strain to eight environmental E. coli isolates: only four isolates displayed adhesion properties to glass significantly higher than PHL565. The ability of the adhesion-proficient isolates to attach to glass tubes strongly correlated with their ability to express curli (thin aggregative fimbriae), thus suggesting that curli are a common adhesion determinant in environmental strains. Despite its inability to attach to solid surfaces, growth of E. coli PHL565 in mixed cultures with Pseudomonas putida MT2 resulted in co-adhesion and in formation of a mixed E. coli/P. putida biofilm, which was able to colonize glass surfaces with dramatic efficiency compared to P. putida alone. E. coli/P. putida interactions stimulate initial adhesion to glass, and the presence of both bacterial species in the mature biofilm was confirmed by quantitative PCR. In contrast, no synergistic biofilm formation was observed in mixed cultures of E. coli with the Gram-positive bacterium Staphylococcus epidermidis. Interestingly, E. coli PHL565 also stimulated biofilm formation by bacterial communities isolated from drinking water distribution systems. Our results strongly suggest that co-adhesion and synergistic interaction with biofilm-forming species might represent an important mechanism, and a possible alternative strategy to production of adhesion determinants, for persistence and propagation of E. coli in the environment.

Filamentous Alphaproteobacteria associated with bulking in industrial wastewater treatment plants.

The phylogeny and distribution of filamentous Alphaproteobacteria, morphologically similar to "Nostocoida limicola" and Eikelboom Type 021N that cause the solids separation problem of bulking in industrial activated sludge plants is described here. A combination of culture-dependent and culture-independent molecular methods has characterized 5 novel species. 16S rRNA targeted oligonucleotide probes were designed for their in situ identification by fluorescence in situ hybridisation (FISH) and used to monitor their presence in 86 WWTPs treating different industrial effluents in four European countries. The involvement of these bacteria in bulking in these plants was confirmed. Filaments hybridising with the ALF-968 probe for the Alphaproteobacteria were present in 65% of the WWTPs examined. They were dominant and therefore probably responsible for bulking in 25.5% of them. The heterogeneous filamentous alphaproteobacterial populations in these communities could be completely identified after application of the oligonucleotide probes used in this study in 91% of the plants containing them. The only filamentous Alphaproteobacteria retrieved in pure culture was isolated from three different industrial WWTPs plants. None of these isolates could grow anaerobically on glucose or denitrify, but all grew aerobically and heterotrophically on a range of carbon sources. Although morphologically similar to the Eikelboom Type 021N morphotype, they were not involved in sulphur metabolism. These bacteria accumulated lipidic storage granules that were associated with their presence under the unbalanced growth conditions existing in these plants.