Effect Of Pretreatment Research Articles

Provenance and pretreatment effect on seed germination of twenty-six provenances of Xanthoceras sorbifolium

ABSTRACT Due to the long dormancy period and hard seed coat, the natural germination percentage of yellowhorn (Xanthoceras sorbifolium) seeds is low, which seriously hinders the large-scale cultivation of the yellowhorn. In this study, the seeds of 26 provenances with different climatic and geographical conditions were collected and the seed germination was tested, which was conducted to determine the effects of seed pretreatment methods on the germination of yellowhorn. This study showed that the germination percentage (GP), mean germination time (MGT) and germination index (GI) of 26 provenances of yellowhorn seeds in different pretreatment methods were highly significant (p < 0.01). The sand storage + clean water and sand storage + PEG almost significantly promote the GP and GI, and reduce the MGT of all provenances, while the pretreatments of warm water and cold water have almost no promoting effect on the GP and GI. In addition, this study indicates that provenances are as important as pretreatments in germination ability. Correlation between germination ability and geo-climatic data of provenances showed that low temperature can improve seed germination ability. This study showed that the seed germination ability of yellowhorn can be improved by using combined pretreatments for provenances with genetically low germination potential.

Effect of ultrasonic pretreatment with synergistic microbial fermentation on tenderness and flavor of air-dried duck under low nitrite process

Effect of ultrasonic pretreatment with synergistic microbial fermentation on tenderness and flavor of air-dried duck under low nitrite process

Promoted lignocellulose fractionation and improved enzymatic hydrolysis of corn stalks through cationic surfactant combined with deep eutectic solvent pretreatment.

The efficient conversion of lignocellulose relies on the implementation of an effective pretreatment strategy. Cationic surfactants combined with deep eutectic solvent ([Betaine][LA] was employed as a novel pretreatment strategy for treating corn stalks. Valuable insights were provided on the impact of the surfactant hydrophobic segment length on pretreatment efficacy. The specific pretreatment conditions were optimized by single factor and orthogonal experiment. Octadecyl trimethyl ammonium bromide (OTAB, 1.5 wt%) with the longest hydrophobic part combined with [Betaine][LA] (Betaine-to-LA molar ratio 1:4) achieved the best pretreatment effect (delignification 92.8 %, xylan elimination 91.1 %) when severity factor reached 4.26, meanwhile, 1.7 g/L xylo-oligosaccharides and 4.4 g/L furfural were detected in pretreatment liquid due to the hydrolysis of hemicellulose in corn stalks with acidic deep eutectic solvent [Betaine][LA]. The relative saccharification activity reached 2.7 times of raw material, while biomass crystallinity, lignin surface area and lignin hydrophobicity significantly decreased, leading to enhanced cellulose accessibility. Additionally, molecular perspective provided by molecular dynamics showed the elimination of lignin and hemicellulose was facilitated by strong interaction of hydrogen-bond and van der Waals force between lignin and hemicellulose with [Betaine][LA] + OTAB. Overall, the effectiveness and potential of cationic surfactant combined deep eutectic solvent pretreatment strategy for lignocellulose pretreatment was revealed.

Effect of the Total Saponins of Bupleurum chinense DC. Water Extracts Following Ultrafiltration Pretreatment on Macroporous Resin Adsorption

Macroporous resin is an efficient separation technology that plays a crucial role in the separation and purification of traditional Chinese medicine (TCM). However, the application of macroporous resins in TCM pharmaceuticals is hindered by serious fouling caused by the complex materials used in TCM. This study examines the impact of ultrafiltration (UF) membrane technology on the macroporous resin adsorption behavior of TCM extracts. In this paper, Bupleurum chinense DC. (B. chinense) water extracts were included as an example to study the effect of UF pretreatment on the macroporous resin adsorption of total saponins. The study results indicated that the adsorption of total saponins constituents from the water extracts of B. chinense on the macroporous resin followed the pseudo-second-order kinetic model and the Langmuir model. The thermodynamic parameters of adsorption, including enthalpy changes and Gibbs free energies, were negative, while entropy changes were positive. These results demonstrated that the total saponin components form a monolayer adsorption layer by spontaneous thermal adsorption on the macroporous resin, and that the adsorption rate is not determined by the rate of intraparticle diffusion. Following treatment with a UF membrane with an average molecular weight cut-off of 50 kDa, the protein, starch, pectin, tannin, and other impurities in the water extracts of B. chinense were reduced, while the total saponin content was retained at 82.32%. The adsorption kinetic model of the saponin constituents on the macroporous resin remained unchanged and was consistent with both the second-order kinetic model and the Langmuir model; the adsorption rate of the second-order kinetic model increased by 1.3 times and in the Langmuir model at 25 °C, the adsorption performance improved by 1.16 times compared to the original extracts. This study revealed that UF technology as a pretreatment method can reduce the fouling of macroporous resin by TCM extracts and improve the adsorption performance of macroporous resin.

Effects of NaOH and Na2CO3 pretreatment on the saccharification of sweet sorghum bagasse

In this work, the chemical composition, chemical structures, and sugar release were evaluated for sweet sorghum bagasse (SSB) biomass in response to alkaline pretreatments and enzymatic hydrolysis. Five different ratios of 2 M sodium hydroxide (NaOH) and sodium carbonate (Na2CO3) solutions were used for biomass fractionation along with two heat treatment conditions, 85°C and 150°C for 1 h. Sugar yields from enzymatic hydrolysis were measured at different durations up to 72 h. SSB samples pretreated at high temperature using alkaline solutions containing equal amounts of Na2CO3 and NaOH favored retaining cellulose content (up to 90%) while effectively removing hemicellulose (65%) and lignin (92%). The high-temperature pretreatment conditions resulted in improved delignification, higher sugar concentrations, and sugar yields. Pretreatment solutions consisting of 50% or more NaOH produced higher glucose yields (85%–90%) and total sugar concentrations. Moreover, pretreatment solutions containing Na2CO3 improved the hydrolysis rates allowing SSB samples reach maximum yields faster than those without it, 48 versus 72 h. Results showed pretreatment methods combining NaOH and Na2CO3 were effective at promoting SSB biomass conversion, increasing sugar recovery after hydrolysis, and reducing the hydrolysis duration, all of which are desirable and cost beneficial.

Enhanced anaerobic digestion of freezing and thawing pretreated cow manure with increasing solid content: kinetics and microbial community dynamics

High solid anaerobic digestion has proved the mainstream technology for the treatment of organic wastes. However, the high molecular weight and complex lignocellulosic structure of cow manure (CM) make it indigestible and inefficient, leading to limit the hydrolysis step of anaerobic digestion at high solid content. To mitigate this bottleneck, an improved cost-effective freezing and thawing pretreatment technique was proposed in this study. The freezing and thawing pretreatment of raw CM without any dilution was done for 20 days. The maximum cumulative methane yield (487 mL CH4 g− 1VS) was achieved at a total solid (TS) of 5% followed by TS of 10% and 15%, which was 13%, 20% and 21% higher than obtained from untreated CM, respectively. The kinetic results revealed that the biodegradable materials could be utilized at increasing TS with decreasing hydrolysis rate. The pretreatment significantly enhanced the methylotrophic methanogenic pathway during high solid anaerobic digestion, which was contrary to the general concept that the process is usually dominated by acetoclastic and hydrogenotrophic methanogens. This study is very important to understand the effect of solid content but also important to understand the effect of freezing and thawing pretreatment on process parameters and microbial community dynamics in high solid anaerobic digestion.

Protective Effect of Protocatechuic Aldehyde on Cerebral Ischemia/Reperfusion Injury in Rats through Blood-Brain Barrier Protection.

Cerebral ischemia can lead to destruction of the blood-brain barrier (BBB), the main cause of cerebral edema and cerebral infarction. BBB damage is also one of the key factors affecting the result of drug therapy. We studied the protective effect of 5-day pretreatment with protocatechuic aldehyde (PAL) at doses of 10 and 20 mg/kg on BBB function and structure after middle cerebral artery occlusion/reperfusion (MCAO/R) in rats. The infarct volume, behavioral neurological deficit score, and Evans blue content in the brain were estimated. We also evaluated the content of nitric oxide (NO) and activities of inducible and neuronal NO synthases. Expression of aquaporin-4 (AQP-4), occludin, claudin-5, and MMP-3 in the brain tissues was estimated by Western blotting. The BBB ultrastructure was analyzed under an electron microscope. We revealed that PAL at both used doses significantly reduced the neurological deficit score, brain infarct volume, and Evans blue extravasation. Electron microscopy showed that PAL significantly improved the ultrastructure of BBB and alleviated its injury. Pretreatment with PAL increased expression of occludin and claudin-5 and reduced expression of AQP-4 and MMP-3. At the same time, the release of NO and activities of NO synthases were notably inhibited. Our results suggest that PAL can be a promising compound to attenuate cerebral ischemia resulting from occlusion/reperfusion injury via BBB protection.

Effect of Different Chemical Pretreatments and Packaging Systems on Quality Characteristics of Fresh-Cut Pomegranate Arils (Punica granatum, L.)

Aims: Pomegranate (Punica granatum) is considered as challenging to consume due to its hard structure, hence used for fresh-cut production. Extending the shelf life of fresh-cut pomegranate arils would improve their marketability, transport efficiency, and reduce post-harvest losses. Study Design: Completely Randomized Design by using GRAPES (General R based Analysis Platform Empowered by Statistics). Place and Duration of Study: College of Agriculture, Kerala Agricultural University, February 2022-July 2024. Methodology: Pomegranate arils were extracted and treated with different pre-treatment solutions viz., 0.5% ascorbic acid, 0.5% citric acid, 1% calcium chloride and 1% calcium ascorbate for 4 minutes. The best pre-treatment solution was selected and arils were packaged in different packaging systems viz., shrink wrapping in 15µ polyolefin film, vacuum packaging in laminated pouches, MAP with N2 flushing in laminated pouches, aluminium tray wrapped with cling film and stored under two different storage conditions, after treating with 1% calcium ascorbate. Results: 1% calcium ascorbate was selected as the best pre-treatment solution and pre-treated arils kept in MAP with N2 flushing in laminated pouches stored under low temperature condition was found to be the best packaging and storage system. 1% calcium ascorbate treated arils in MAP with N2 flushing in laminated pouches stored under low temperature condition (10±5oC) had a shelf life of 6 days with superior physiological (PLW, percent leakage), chemical (acidity, vitamin C, phenol) and sensory characters (color, flavour, texture, appearance, taste and overall acceptability) as compared to control (2.00 days). Maximum vitamin C content of 23.02 mg 100g-1 and least phenol content of 214.16 mg 100g-1 were recorded on 6th day of storage. Conclusion: Pomegranate arils pre-treated with 1% calcium ascorbate for 4 minutes, kept in MAP with N2 flushing in laminated pouches stored under low temperature condition (10±5oC) can be considered as a protocol for shelf life extension of pomegranate arils.

Significantly enhancing enzymatic saccharification of poplar waste through the effective removal of lignin and hemicellulose by 1-butanol-Na3PO4-water treatment

Na3PO4-H2O-assisted 1-butanol pretreatment was established for changing the structure of poplar waste and improving its enzymatic efficiency. After poplar waste was pretreated at 180 °C for 60 min, 51.3 % of xylan and 52.5 % of lignin were eliminated. After the enzymatic saccharification of treated poplar waste, the yield of reducing sugars reached 75.3 %. A series of characterizations demonstrated that Na3PO4-H2O-assisted 1-butanol pretreatment altered the poplar composition with improved cellulose accessibility from 231.2 to 459.5 mg/g and reduced surface area of lignin from 384.2 to 255.3 m2/g. Molecular simulations, quantum chemical calculations and independent gradient model analysis were performed on the poplar model, and the 1-butanol-Na3PO4-H2O pretreatment resulted in the increase of electrostatic and van der Waals repulsion forces in lignin and xylan, explaining the efficient lignin and xylan removal. A comprehensive interpretation of mass balance and energy flow was applied to assess the entire pretreatment process. This eco-friendly 1-butanol-Na3PO4-H2O pretreatment effectively broken the encapsulation of cellulose by enzyme inhibitors and improve biomass pretreatment effect.

Clinical outcomes of round spermatid injection versus intra-cytoplasmic sperm injection: The potential role of hormonal pretreatment in male patients with non-obstructive azoospermia.

Round spermatid injection (ROSI) into oocytes offers men with non-obstructive azoospermia (NOA) the opportunity to have biological offspring in cases where mature spermatozoa are not detected. However, the clinical outcomes of ROSI remains poor. This study compares the outcomes of ROSI with intracytoplasmic sperm injection (ICSI) and investigates the effect of hormonal pretreatment. This retrospective cohort study enrolled infertile couples undergoing either ROSI or ICSI at the reproductive center in Taipei Veterans General Hospital. The administration of selective estrogen receptor modulators, gonadotropins, and aromatase inhibitors in male patients were collected. Relevant hormonal markers and biochemical parameters were determined. The outcomes of ROSI and ICSI were assessed based on fertilization rate, implantation rate, and live birth rate. A total of 36 couples were recruited in the ROSI group, whereas 39 couples were recruited in the ICSI group for the analysis. Patients in each group demonstrated similar characteristics, except for a higher proportion of male patients in the ROSI group who were pretreated with anastrozole. The fertilization rate and implantation rate were similar between ROSI and ICSI groups after adjusting for confounding variable. The live birth rate was significant lower in the ROSI group (8.3%) than in the ICSI group (30.8%) before and after adjusting for confounding variable. ROSI demonstrated fertilization and implantation rates comparable to those of ICSI for male patient with NOA undergoing testicular sperm extraction surgery. Further studies evaluating the effect of anastrozole administration on ROSI outcomes are warrant.

Effect of Curing Time and Ferric Chloride on a Copper Concentrate with a High Arsenic Content

As a result of changes in copper mineralogy, various treatment options for copper sulfides have been explored, including pretreatment processes aimed at enhancing material permeability and improving the dissolution of valuable minerals. Despite its significance, this topic has only recently gained attention. In this research, a copper concentrate with a high arsenic content was studied, with enargite (Cu3AsS4) as the main mineral phase. The objective was to evaluate the effect of pretreatment on copper extraction efficiency prior to leaching. Three key variables were investigated: curing time (0, 5, 10, and 15 days), H2SO4 dosage (0, 70, 140, and 210 kg/t), and FeCl3 concentration (0, 0.5, 1, and 1.5 M). The sample was characterized both before and after pretreatment, revealing the formation of new species such as CuSO4·5H2O and Cu2Cl(OH)3 under optimal conditions of 15 days curing time, 70 kg/t of H2SO4, and 1 M FeCl3. Copper extraction solely through curing reached 20.79%. The analysis suggests that curing time is the most influential factor in the process, accounting for 46% of the overall contribution. In comparison, sulfuric acid and ferric chloride contribute less, with 20% and 10% contributions, respectively.

Effect of microwave hydrothermal pretreatment on dissolution of composite components in Acacia wood and subsequent pulping performance

Effect of microwave hydrothermal pretreatment on dissolution of composite components in Acacia wood and subsequent pulping performance

Shift of plasmon resonance in silver nanoparticles: effect of magnetic field pre-treatment

Abstract A novel magnetic field induced phenomenon in ZnO/Ag nanoparticles is detected and investigated with reference to the surface plasmon resonance (SPR) effect. A shift of the maximum of plasmon absorption of the ZnO/Ag nanoparticles formed in different days in magnetic field treated ZnO/ethanol solutions was observed. The observed phenomena were explained in terms of the ferromagnetic-like properties of the ZnO nanoparticles due to the surface broken bonds, which result in the appearance of non-zero magnetic moments. Magnetic field pre-treatment may be used as an effective tool for manipulating the plasmon properties of silver-based nanoparticles, which is perspective for creating new-generation sensor systems.

Impact of Nanoparticle Addition and Ozone Pre-Treatment on Mesophilic Methanogenesis in Temperature-Phased Anaerobic Digestion

Biodegradable organic waste offers significant opportunities for resource recovery within the frame of the circular economy. In this work, the effects of carbon-encapsulated iron nanoparticles and ozone pre-treatments in the mesophilic methanogenic stage of a temperature-phased an-aerobic digestion have been studied using biochemical methanogenic potential (BMP) tests and modeling simulation. To do that, digestates from a pre-treated thermophilic acidogenic reactor that co-digested sludge and wine vinasse were used. The addition of nanoparticles favored the removal of particulate matter, which increased by 9% and 6% in terms of total solids and volatile solids, respectively. When combined with ozone pre-treatment, these increases were 27% and 24%, respectively, demonstrating enhanced AD efficiency. The dose of iron nanoparticles encapsulated in carbon did not result in a statistically significant increase in methane production when sludge and vinasse were used as feedstock. The combination of nanoparticles with the ozone pre-treatment significantly improved the methanogenic phase of the second stage, increasing the methane production yield by 22% and reducing the lag phase from 10 days to 3 days, according to the modified Gompertz model.

Enhanced performance and economic feasibility of sewage sludge digestion using a two-stage anaerobic digestion with a dynamic membrane and alkaline-thermal pretreatment

This study suggests a high-rate sewage sludge anaerobic digestion (AD) process. An alkaline-thermal pretreatment and a dynamic membrane (DM) were used to enhance AD efficiency and economic feasibility in a two-stage system. The effect of pretreatment on volatile fatty acid (VFA) production in the acidogenic phase was investigated at various hydraulic retention times (HRT). After optimizing the acidogenic phase condition (HRT of 3 days), single- and two-stage AD processes with DM modules were operated simultaneously to compare performance. The highest methane production rates of 0.69 L/L/d for single-stage AD and 1.10 L/L/d for two-stage AD were observed at a total HRT of 12 days. Phase separation enhanced the growth of acetoclastic methanogens. A techno-economic analysis showed that the two-stage AD system would achieve a positive net present value within 2 years. This study demonstrated the feasibility of high-rate AD systems for sewage sludge using DM, alkaline-thermal pretreatment, and phase separation.

Effect of aeration pretreatment on anaerobic digestion of swine manure

ABSTRACT To investigate the effects of aeration pretreatment on the anaerobic digestion (AD) of swine manure, five pretreatment groups were established with dissolved oxygen (DO) in each group set to 0.0, 0.4, 0.8, 1.4, and 2.0 mg/L, respectively. The results demonstrated that compared to the non-aeration group, methane production increased to varying degrees with different aeration pretreatments (AP), with a maximum increase of 27.98% (DO = 2.0 mg/L). AP reduced the hydrogen sulfide (H2S) content of biogas. The H2S concentration in the DO = 2.0 mg/L was only 0.209%, and this represented an increased H2S removal rate of 49.27% compared to that of the DO = 0.0 mg/L (0.412%). Simultaneously, AP increases the hydrolysis rate. When the DO concentration reached 2.0 mg/L, the hydrolysis rate reached its maximum. An increase in the hydrolysis rate further enhanced the removal rate of organic matter. The organic matter removal rate was highest (36.96%) at DO = 2.0 mg/L. AP effectively prolonged the methane generation time and shortened the lag time of methane generation. AP creates a brief micro aerobic environment, accelerates substrate hydrolysis, and promotes the production and consumption of total volatile fatty acids, particularly acetic acid. Additionally, AP promoted the symbiotic relationship between Caldicoprobacter (20.93%–34.96%) and Metanosaeta (14.73%–18.45%).

GlyT1 inhibition promotes neuroprotection in the middle cerebral artery occlusion model through the activation of GluN2A-containing NMDAR

Glycine Transporter Type 1 (GlyT1) inhibition confers neuroprotection against different forms of cerebral damage. This effect occurs through the elevation of synaptic glycine concentrations, which enhances N-methyl-d-aspartate receptor (NMDAR) activation by glutamate. To investigate the neuroprotective mechanism of GlyT1 inhibition, we used the Middle Cerebral Artery Occlusion (MCAO) model in male C57BL/6 mice, aged 10–12 weeks. We administered N-[3-(4′-fluorophenyl)-3-(4′-phenylphenoxy)propyl] sarcosine (NFPS), a GlyT1 inhibitor, 24 h prior to ischemia induction. NFPS pretreatment provided significant neuroprotection in the MCAO model, associated with modulation of pathways related to long-term potentiation. Specifically, GluN2A subunit expression was upregulated, while GluN2B subunit expression was downregulated in cortical areas, correlating with enhanced phosphorylation of CaMKIV and CREB proteins. Coadministration with the GluN2B antagonist Eliprodil or the CREB inhibitor C646 did not affect the neuroprotective effects of NFPS pretreatment, but TCN-201, a specific GluN2A antagonist, disrupted these effects. These findings suggest that GlyT1 inhibition mediates neuroprotection through activation of GluN2A-containing NMDARs and the GluN2A/CaMKIV/CREB signaling cascade, thereby modulating the balance between GluN2A and GluN2B subunits.

Effect of delignification pretreatment and furfurylation on practical properties of alder wood (Alnus spp.)

Partial delignification of alder wood specimens was performed with alkaline sulfite and peracetic acid (PAA) methods, and subsequently the delignified samples were saturated with furfuryl alcohol, as an eco-friendly chemical. The aim of delignification before furfurylation was to facilitate the penetration of furfuryl alcohol into the cell wall, in order to overcome the weak mechanical properties and hydrophilicity of alder wood as a fast growing species. The results showed that alkaline delignification removed a lower percentage of lignin despite more significant mass loss as compared to peracetic acid delignification method, which indicates more carbohydrates degradation in alkaline delignification. Infrared spectroscopy also confirmed more retention of carbohydrates in the sample delignified with peracetic acid. The polymerization of furfuryl alcohol in the cell cavities decreased the access of water molecules to the hydroxyl groups of the cell wall, increased the contact angle, and decreasing surface wettability. This effect was more noticeable in the PAA-delignified samples. Mechanical tests revealed that the delignified samples with peracetic acid had higher mechanical strength and modulus, compared to the alkaline sulfite delignified samples. This was attributable to the maintenance of carbohydrates during the delignification process.

Change in components by pretreatments alters the antimicrobial activity of Strobilanthes cusia biomass

The plant Strobilanthes cusia contains various antibacterial components, but the mechanisms behind its antimicrobial activity are not yet fully understood. This study demonstrates that alterations in components due to different pretreatments significantly impacted the antimicrobial activity of S. cusia biomass. Biomass dried using microwave pretreatment, which contained a high amount of indican, exhibited the lowest antibacterial activity, whereas heat-dried biomass without indican showed the highest antibacterial activity. High-performance liquid chromatography analysis indicated that pretreatments influenced the ratios of indican, indoxyl, indigo, and indirubin in S. cusia biomass, likely due to the effect of pretreatments on β-glucosidase activity. Extracts with higher levels of indoxyl, an intermediate product of indican hydrolysis, exhibited stronger inhibitory effects against Staphylococcus aureus. Indoxyl increased the NAD+/NADH ratio, which led to elevated hydrogen peroxide generation and subsequent cell damage. Furthermore, transmission electron microscopy, scanning electron microscopy, and confocal laser scanning microscopy revealed distinct morphological changes in cells exposed to indoxyl. Finally, leveraging the water-soluble properties of indoxyl, we propose a novel approach using microwave drying assisted by β-glucosidase to enhance the antimicrobial activity of S. cusia biomass. These findings provide a theoretical and methodological foundation for the evaluation and future application of S. cusia's bioactive substances.

Feasibility of biogas upgrading at a WWTP after pre-treatment application: Techno-economic assessment validation with pilot test data

Improving the efficiency of anaerobic digestion (AD) of sewage sludge (SS) is a critical step toward the achievement of energy neutrality in wastewater treatment plants (WWTPs), as required by the European Green Deal. This study used a comparative techno-economic assessment (TEA) to evaluate the feasibility of producing biomethane, at a WWTP, through upgrading biogas with a double-stage permeation membrane plant. The biogas was originally generated from the AD of a mixture of primary sludge (PS) and either raw or pre-treated waste activated sludge (WAS), where biological or thermo-alkali pre-treatments were applied to increase the WAS intrinsic low degradability.The TEA was supported by the results of pilot-scale tests, carried out on WAS, which mimicked (i) a traditional mesophilic AD process; (ii) a two-stage AD process, where a temperature-phased anaerobic digestion (TPAD, 3 days, 55 °C + 20 days, 38 °C) was performed to biologically pre-treat WAS; (iii) a traditional mesophilic AD process preceded by a thermo-alkali (4 g NaOH/100 g TS, 90 °C, 90 min) pre-treatment.The TEA was carried out in two phases. In the first, the minimum size of the WWTP capable of making the costs necessary for the implementation of the biogas upgrading plant equal to the revenues coming from selling biomethane (at 62 €/MWh) in 10 years was calculated in the absence of pre-treatments. It resulted of 500,000 equivalent inhabitants (e.i.). In the second phase, for the WWTP size found previously, the effect of either biological or thermo-alkali pre-treatments on the economic balance was evaluated, that is the gain (or the loss) associated to the selling of biomethane, compared to the reference price of 62 €/MWh. It was found that the TPAD increased the biogas productivity by only 23.6%, too little to compensate the amount of heat necessary for the pre-treatment and the purchase cost of the additional reactor. Conversely, the thermo-alkali pre-treatment, which enhanced the WAS biogas productivity by 110%, increased the biomethane revenues by approx. 10 €/MWh, compared to the scenario without pre-treatments. This study offers useful data to WWTP managers who want to introduce WAS pre-treatments, combined with interventions for biogas upgrading, in a new or existing sludge line of a WWTP.