Temperature-driven carboxylic acid production from waste activated sludge and food waste: Co-fermentation performance and microbial dynamics.

This work aims to improve the continuous co-fermentation of waste activated sludge (WAS) and food waste (FW) by investigating the long-term impact of temperature on fermentation performance and the underpinning microbial community. Acidogenic co-fermentation of WAS and FW (70:30 % VS-basis) to produce volatile fatty acids (VFA) was studied in continuous fermenters at different temperatures (25, 35, 45, 55 °C) at an organic loading rate of 11 gVS/(L·d) and a hydraulic retention time of 3.5 days. Two batches of WAS (A and B) were collected from the same wastewater treatment plant at different periods to understand the impact of the WAS microbioota on the fermenters' microbial communities. Solubilisation yield was higher at 45 °C (575 ± 68 mgCOD/gVS) followed by 55 °C (508 ± 45 mgCOD/gVS). Fermentation yield was higher at 55 °C (425 ± 28 mgCOD/gVS) followed by 35 °C (327 ± 17 mgCOD/gVS). Temperature also had a noticeable impact on the VFA profile. At 55 °C, acetic (40 %) and butyric (40 %) acid dominated, while acetic (37 %), butyric acid (31 %), and propionic acid (17 %) dominated at 35 °C. At 45 °C, an accumulation of caproic acid was detected which did not occur at other temperatures. Each temperature had a distinct microbial community, where the WAS microbiota played an important role. The biomass mass-balance showed the highest growth of microorganisms (51 %) at 35 °C and WAS_B, where a consumption of acetic acid was observed. Therefore, at 35 °C, there is a higher risk of acetic acid consumption probably due to the proliferation of methanogens imported from WAS.

Fractional calculus as a generalized kinetic model for biochemical methane potential tests.

This study presents a fractional calculus model as a generalized kinetic model for estimating the maximum methane yield and degradation kinetics in biomethane potential (BMP) assays, a key analytical method in anaerobic digestion research and application. The fractional model outperformed common first-order kinetic models by yielding superior data fitting and properly managing substrate heterogeneity. The fractional model showed robust performance in mono-digestion, co-digestion and pre-treatment BMP assays with or without presence of large tailing or sigmoidal patterns in the BMP curve. The main advantage of the fractional model over other models is its ability to capture the complexities of the methane production process without losing model accuracy. Assessment of the mathematical model revealed that for fractional orders greater than 0.8 the Mittag-Leffler sequence could be transformed into a more computationally efficient exponential function.

Impact of food waste composition on acidogenic co-fermentation with waste activated sludge.

The impact of food waste (FW) composition on co-fermentation performance was studied to elucidate if adjusting FW composition can be used to drive the fermentation yield and profile, which is relevant for biorefinery applications. First, the impact of individual FW components (i.e., fruit, vegetables, pasta, rice, meat, fish, and cellulose) was assessed. Subsequently, the effect of mixing a protein-rich component and a carbohydrate-rich component was studied (i.e., fish/fruit and fish/cellulose, and meat/rice and meat/vegetable). All experiments were carried out in mesophilic batch assays using waste activated sludge (WAS) as main substrate, the same mixture ratio (70 % WAS +30 % FW on VS basis), and no pH control. Results showed that each FW component had a distinct effect on VFA yield and profile, with protein-rich components reaching the highest VFA yields; 502 and 442 mgCOD/gVS for WAS/Fish and WAS/Meat, respectively. A positive interaction on VFA yield was observed when mixing a protein-rich and a carbohydrate-rich component. This interaction was not proportional to the co-substrates proportion in the mixtures. On the other hand, the VFA profile was clearly driven by the components in the mixture, including both WAS and FW composition. Overall, these results indicate that predicting the VFA yield of WAS/FW co-fermentation is not just related to FW composition, but FW composition could be used to adjust the VFA profile to a certain extent.

Gamma distribution function to understand anaerobic digestion kinetics: Kinetic constants are not constant.

The Gamma model is a novel approach to characterise the complex degradation dynamics taking place during anaerobic digestion. This three parameters model results from combining the first-order kinetic model and the Gamma distribution function. In contrast to conventional models, where the kinetic constant is considered invariant, the Gamma model allows analysing the variability of the kinetic constant using a probability density function. The kinetic constant of mono-digestion and co-digestion batch tests of different wastes were modelled using the Gamma model and two common first-order models: one-step one-fraction model and one-step two-fraction model. The Gamma distribution function approximates three distinct probability density functions, i.e. exponential, log-normal, and delta Dirac. Specifically, (i) cattle paunch and pig manure approximated a log-normal distribution; (ii) cattle manure and microalgae approximated an exponential distribution, and (iii) primary sludge and cellulose approximated a delta Dirac distribution. The Gamma model was able to characterise two distinct waste activated sludge, one approximated to a log-normal distribution and the other to an exponential distribution. The same cellulose was tested with two different inocula; in both tests, the Gamma distribution function approximated a delta Dirac function but with a different kinetic value. The potential and consistency of Gamma model were also evident when analysing pig manure and microalgae co-digestion batch tests since (i) the mean k of the co-digestion tests were within the values of the mono-digestion tests, and (ii) the profile of the density function transitioned from log-normal to exponential distribution as the percentage of microalgae in the mixture increased.

The novel genus, 'Candidatus Phosphoribacter', previously identified as Tetrasphaera, is the dominant polyphosphate accumulating lineage in EBPR wastewater treatment plants worldwide.

The bacterial genus Tetrasphaera encompasses abundant polyphosphate accumulating organisms (PAOs) that are responsible for enhanced biological phosphorus removal (EBPR) in wastewater treatment plants. Recent analyses of genomes from pure cultures revealed that 16S rRNA genes cannot resolve the lineage, and that Tetrasphaera spp. are from several different genera within the Dermatophilaceae. Here, we examine 14 recently recovered high-quality metagenome-assembled genomes from wastewater treatment plants containing full-length 16S rRNA genes identified as Tetrasphaera, 11 of which belong to the uncultured Tetrasphaera clade 3. We find that this clade represents two distinct genera, named here Ca. Phosphoribacter and Ca. Lutibacillus, and reveal that the widely used model organism Tetrasphaera elongata is less relevant for physiological predictions of this uncultured group. Ca. Phosphoribacter incorporates species diversity unresolved at the 16S rRNA gene level, with the two most abundant and often co-occurring species encoding identical V1-V3 16S rRNA gene amplicon sequence variants but different metabolic capabilities, and possibly, niches. Both Ca. P. hodrii and Ca. P. baldrii were visualised using fluorescence in situ hybridisation (FISH), and PAO capabilities were confirmed with FISH-Raman microspectroscopy and phosphate cycling experiments. Ca. Phosphoribacter represents the most abundant former Tetrasphaera lineage and PAO in EPBR systems in Denmark and globally.

Potential of anaerobic co-fermentation in wastewater treatments plants: A review.

Fermentation (not anaerobic digestion) is an emerging biotechnology to transform waste into easily assimilable organic compounds such as volatile fatty acids, lactic acid and alcohols. Co-fermentation, the simultaneous fermentation of two or more waste, is an opportunity for wastewater treatment plants (WWTPs) to increase the yields of sludge mono-fermentation. Most publications have studied waste activated sludge co-fermentation with food waste or agri-industrial waste. Mixing ratio, pH and temperature are the most studied variables. The highest fermentation yields have been generally achieved in mixtures dominated by the most biodegradable substrate at circumneutral pH and mesophilic conditions. Nonetheless, most experiments have been performed in batch assays which results are driven by the capabilities of the starting microbial community and do not allow evaluating the microbial acclimation that occurs under continuous conditions. Temperature, pH, hydraulic retention time and organic load are variables that can be controlled to optimise the performance of continuous co-fermenters (i.e., favour waste hydrolysis and fermentation and limit the proliferation of methanogens). This review also discusses the integration of co-fermentation with other biotechnologies in WWTPs. Overall, this review presents a comprehensive and critical review of the achievements on co-fermentation research and lays the foundation for future research.

Assessing the potential of waste activated sludge and food waste co-fermentation for carboxylic acids production.

This study investigated waste activated sludge (WAS) and food waste (FW) co-fermentation in batch assays to produce carboxylic acids. Three mixtures (50%, 70% and 90% WAS in VS basis) were studied under different conditions: with and without extra alkalinity, and with and without WAS auto-hydrolysis pre-treatment. All tests were carried out at 35 °C, without pH adjustment and without external inoculum. Experimental results showed that co-fermentation yields, including volatile fatty acids and lactic acid, were always higher than WAS and FW mono-fermentation yields (ca. 100 and 80 mgCOD/gVS, respectively). Co-fermentation yields increased as the proportion of FW in the mixture increased, indicating that the improvement was primarily due to a higher FW degradation under co-fermentation conditions. The maximum co-fermentation yield was on average 480 mgCOD/gVS for the WAS/FW_50/50 mixture. The importance of pH on co-fermentation performance was evident in the experiments carried out with extra alkalinity, which showed that the proportion of WAS in the mixture should be high enough to keep the pH above 5.0. However, fermenters operational conditions should also prevent the enrichment of acetic acid consuming microorganisms. WAS auto-hydrolysis pre-treatment did not enhance co-fermentation yields but showed minor kinetic improvements. Regarding the product profile, butyric acid was enriched as the proportion of FW in the mixture increased and the concomitant pH decreased to the detriment of propionic acid. Propionic acid prevailed under neutral pH in the WAS mono-fermentation and the WAS/FW_90/10 mixture.

Humic acid inhibition of hydrolysis and methanogenesis with different anaerobic inocula.

There is increasing evidence that humic acid (HA) is hampering the performance of anaerobic digesters treating animal manures and thermally-hydrolysed waste activated sludge. In the present study, HA inhibition and inhibition resilience was examined for hydrolysis (carbohydrate and protein) and acetotrophic methanogenesis with four distinct full-scale anaerobic inocula. The aim was to further understand HA inhibition and to explore potential relationships between microbial factors and inhibition resilience. For two of the four tested inocula, cellulose degradation showed a start-up delay that lengthened as HA concentration increased from 0 to 2 g L-1. This inhibition was reversible because, after the initial delay, subsequent hydrolysis rates and methane yields were not significantly influenced by HA concentration. Cellulose hydrolysis results at HA concentrations below 2 g L-1 support a threshold inhibition mechanism, i.e. HA complexes with hydrolytic enzymes preventing them from binding with cellulose, but once all the HA had been complexed, enzymes subsequently released are free to bind with cellulose. Inocula with higher cellulose hydrolytic activity were less affected by HA inhibition, suggesting a potential link between HA inhibition resilience and microbial activity. However, above 5 gHA L-1, cellulose hydrolysis rates decreased with increasing HA concentration; indicating that the mechanisms of inhibition may change depending on some threshold HA concentration. Protein hydrolysis and acetotrophic methanogenesis were less susceptible to HA inhibition than cellulose hydrolysis, since signs of inhibition were only observed above 5 gHA L-1. Acetotrophic methanogenesis was partially inhibited at 10 gHA L-1 and completely inhibited at 20 gHA L-1. These results further support that HA inhibition is selective towards particular enzymes.

Characterising and modelling free ammonia and ammonium inhibition in anaerobic systems.

Inhibition by ammoniacal nitrogen, consisting of free ammonia (NH3) and ammonium ion (NH4+), has been widely investigated for anaerobic digestion. However, despite the large amount of research on the subject, ammoniacal nitrogen inhibition still threatens many anaerobic digesters. This paper presents (i) a method to reliably characterise ammoniacal nitrogen inhibition and (ii) a robust inhibition modelling approach. Results showed that NH3 and NH4+ inhibition need to be jointly determined, which can only be done by performing inhibition tests at various total ammoniacal nitrogen (TAN) concentrations and pH values. These test conditions were reliably achieved using the salts NH4HCO3 and NH4Cl without pH adjustment, rather than by using NH4Cl with pH adjustment. The use of only salts showed a lower pH change during the inhibition test (∼1.5 days), thereby decreasing the uncertainty in TAN speciation and strengthening the test and model outputs. A threshold inhibition function satisfactorily described (R2 > 0.99) the joint inhibition of NH3 and NH4+ on three distinct inocula, and provided a better description of the inhibition testing results than a non-competitive inhibition function (R2 ∼0.70). The key advantage of the proposed threshold inhibition function is its capacity to identify the inhibition lower limit (concentration where inhibition starts; KImin) and upper limit (concentration where inhibition is complete; KImax). The threshold inhibition function also identifies the 50% inhibition concentration (KI50) at the midpoint of KImin and KImax. Finally, experimental and model results show that at pH 7.3-7.7 and TAN concentrations above 2000 mgN·L-1, both NH3 and NH4+ contribute significantly to overall inhibition.

Deterministic mechanisms define the long-term anaerobic digestion microbiome and its functionality regardless of the initial microbial community.

The impact of the starting inoculum on long-term anaerobic digestion performance, process functionality and microbial community composition remains unclear. To understand the impact of starting inoculum, active microbial communities from four different full-scale anaerobic digesters were each used to inoculate four continuous lab-scale anaerobic digesters, which were operated identically for 295 days. Digesters were operated at 15 days solid retention time, an organic loading rate of 1 g COD Lr-1 d-1 (75:25 - cellulose:casein) and 37 °C. Results showed that long-term process performance, metabolic rates (hydrolytic, acetogenic, and methanogenic) and microbial community are independent of the inoculum source. Digesters process performance converged after 80 days, while metabolic rates and microbial communities converged after 120-145 days. The convergence of the different microbial communities towards a core-community proves that the deterministic factors (process operational conditions) were a stronger driver than the initial microbial community composition. Indeed, the core-community represented 72% of the relative abundance among the four digesters. Moreover, a number of positive correlations were observed between higher metabolic rates and the relative abundance of specific microbial groups. These correlations showed that both substrate consumers and suppliers trigger higher metabolic rates, expanding the knowledge of the nexus between microorganisms and functionality. Overall, these results support that deterministic factors control microbial communities in bioreactors independently of the inoculum source. Hence, it seems plausible that a desired microbial composition and functionality can be achieved by tuning process operational conditions.

Biochemical methane potential (BMP) tests: Reducing test time by early parameter estimation.

Biochemical methane potential (BMP) test is a key analytical technique to assess the implementation and optimisation of anaerobic biotechnologies. However, this technique is characterised by long testing times (from 20 to >100days), which is not suitable for waste utilities, consulting companies or plants operators whose decision-making processes cannot be held for such a long time. This study develops a statistically robust mathematical strategy using sensitivity functions for early prediction of BMP first-order model parameters, i.e. methane yield (B0) and kinetic constant rate (k). The minimum testing time for early parameter estimation showed a potential correlation with the k value, where (i) slowly biodegradable substrates (k≤0.1d-1) have a minimum testing times of ≥15days, (ii) moderately biodegradable substrates (0.1

Semi-aerobic fermentation as a novel pre-treatment to obtain VFA and increase methane yield from primary sludge.

There is a growing trend to consider organic wastes as potential sources of renewable energy and value-add products. Fermentation products have emerged as attractive value-add option due to relative easy production and broad application range. However, pre-fermentation and extraction of soluble products may impact down-stream treatment processes, particularly energy recovery by anaerobic digestion. This paper investigates primary sludge pre-fermentation at different temperatures (20, 37, 55, and 70°C), treatment times (12, 24, 48, and 72h), and oxygen availability (semi-aerobic, anaerobic); and its impact on anaerobic digestion. Pre-fermentation at 20 and 37°C succeeded for VFA production with acetate and propionate being major products. Pre-fermentation at 37, 55, and 70°C resulted in higher solubilisation yield but it reduced sludge methane potential by 20%. Under semi-aerobic conditions, pre-fermentation allowed both VFA recovery (43gCODVFAkg(-1)VS) and improved methane potential. The latter phenomenon was linked to fungi that colonised the sludge top layer during pre-fermentation.

Effect of moisture on pretreatment efficiency for anaerobic digestion of lignocellulosic substrates.

The present study evaluates the effect of moisture in low-temperature and ultrasound pretreatment on lignocellulosic substrates anaerobic biodegradability, where brewer's spent grain was used as model substrate. Besides moisture content, low-temperature pretreatment was also evaluated in terms of temperature (60-80°C) and exposure time (12-72 h). Likewise, ultrasonication was also evaluated in terms of specific energy (1000-50,000 kJ kg TS(-1)). In addition, the effect of substrate particle size reduction by milling pretreatment was also considered. The results clearly demonstrated that substrate moisture (total solid concentration) is a significant parameter for pretreatment performance, although it has been rarely considered in pretreatment optimisation. Specifically, moisture optimisation increased the methane yield of brewer's spent grain by 6% for low-temperature pretreatment (60°C), and by 14% for ultrasound pretreatment (1000 kJ kg TS(-1)) towards the control (without pretreatment). In both pretreatments, the experimental optimum total solid concentration was 100 gTS kg(-1). Thus, lowering substrate moisture, a strategy suggested attaining energetic pretreatment feasibility, needs to be analysed as another pretreatment variable since it might have limited correlation. Finally, a preliminary energetic balance of the pretreatments under study showed that the extra methane production could not cover the energetic pretreatment expenses.

Assessing total and volatile solids in municipal solid waste samples.

Municipal solid waste is broadly generated in everyday activities and its treatment is a global challenge. Total solids (TS) and volatile solids (VS) are typical control parameters measured in biological treatments. In this study, the TS and VS were determined using the standard methods, as well as introducing some variants: (i) the drying temperature for the TS assays was 105°C, 70°C and 50°C and (ii) the VS were determined using different heating ramps from room tempature to 550°C. TS could be determined at either 105°C or 70°C, but oven residence time was tripled at 70°C, increasing from 48 to 144 h. The VS could be determined by smouldering the sample (where the sample is burnt without a flame), which avoids the release of fumes and odours in the laboratory. However, smouldering can generate undesired pyrolysis products as a consequence of carbonization, which leads to VS being underestimated. Carbonization can be avoided using slow heating ramps to prevent the oxygen limitation. Furthermore, crushing the sample cores decreased the time to reach constant weight and decreased the potential to underestimate VS.

Balancing hygienization and anaerobic digestion of raw sewage sludge.

The anaerobic digestion of raw sewage sludge was evaluated in terms of process efficiency and sludge hygienization. Four different scenarios were analyzed, i.e. mesophilic anaerobic digestion, thermophilic anaerobic digestion and mesophilic anaerobic digestion followed by a 60 °C or by an 80 °C hygienization treatment. Digester performance (organic matter removal, process stability and biogas yield) and the hygienization efficiency (reduction of Escherichia coli, somatic coliphages and F-specific RNA phages) were the main examined factors. Moreover, a preliminary economical feasibility study of each option was carried out throughout an energy balance (heat and electricity). The obtained results showed that both thermophilic anaerobic digestion and mesophilic anaerobic digestion followed by a hygienization step were able to produce an effluent sludge that fulfills the American and the European legislation for land application. However, higher removal efficiencies of indicators were obtained when a hygienization post-treatment was present. Regarding the energy balance, it should be noted that all scenarios have a significant energy surplus. Particularly, positive heat balances will be obtained for the thermophilic anaerobic digestion and for the mesophilic anaerobic digestion followed by 60 °C hygienization post-treatment if an additional fresh-sludge/digested sludge heat exchanger is installed for energy recovery.

The posterior segment of the temporomandibular joint capsule and its anatomic relationship.

PURPOSE: The aim of this work was to clarify the arrangement of the posterior segment of the temporomandibular joint capsule and its pertinent relationships. MATERIALS AND METHODS: The temporomandibular region was dissected bilaterally in 20 adult cadavers. Natural stained latex was injected into 16 cadavers through the external carotid artery to facilitate the dissection of the arterial vessels. RESULTS: The posterior segment of the joint capsule is made up of the so-called "bilaminar zone" of the articular disc. The upper internal portion of the posterior segment of the capsule was reinforced by the discomalleolar ligament. The retroarticular space was filled with loose connective tissue and the anterior branches of the anterior tympanic artery were distributed throughout the posterior segment of the joint capsule. CONCLUSION: The posterior segment of the temporomandibular joint capsule corresponds to the bilaminar zone of the articular disc. The structures of the retroarticular space are extracapsular.

[Functional laryngeal obstruction]. / Obstrucción laríngea funcional.

We describe 4 patients with reversible obstruction of the upper airways, a condition that usually presents with crises of dyspnea and respiratory sounds. No organic cause could be identified and the symptoms were initially confused with those of bronchial asthma. During episodes, the flow-volume curve suggested obstruction of the upper airways with poor reproducibility and repeatedly normal resistance values. Fiberoptic bronchoscopy allowed us to confirm the functional nature of the obstruction in these cases.