Biochemical Methane Potential Tests Research Articles

Impacts of free nitrous acid on stabilizing food waste and sewage sludge for anaerobic digestion

This work investigated the effectiveness of free nitrous acid (FNA) in enhancing organic waste solubilization to improve biogas production in anaerobic digestion (AD). The results indicated that FNA pretreatment can enhance soluble organic content and control H2S odor in tested organic wastes, including food waste, sewage sludge, and their combination. However, a significant decrease (>50 %) in FNA concentration was found in the reactors, possibly due to denitrifier-driven NO2– consumption. Biochemical methane potential (BMP) tests showed a 25 ± 8 % enhancement in CH4 production in the reactors fed with mixed substrate pretreated with 2.9 mg FNA-N/L. However, the presence of NO2– (325.6–2368.0 mg N/L) in some BMP reactors, due to carryover from FNA pretreatment, adversely affected CH4 production (>55 %) and prolonged lag time (>4.2 times). These findings are valuable for researchers and practitioners in waste management, offering insights for implementing FNA pretreatment to enhance the biodegradability of organic wastes in AD.

Mechanisms of metabolic regulation and enhanced methane production by hydrolytic nanozyme in sludge anaerobic digestion

Artificial nanozymes have demonstrated potential as natural enzyme substitutes due to their efficient biomacromolecule hydrolysis. However, the metabolic and catalytic mechanisms of nanozyme during the biodegradation of complex organics remain inadequately understood. This study evaluated the effect of the hydrolytic mechanism of cerium-based metal–organic frameworks (Ce-MOFs) hydrolytic nanozyme on shaping microbiome and metabolic functions. Biochemical methane potential test revealed that Ce-MOF-based hydrolytic nanozyme increased methane production by 7.8 − 22.2% as the hydrolytic nanozyme dosage increased from 50 to 150 mg/g VS. The highest methane production observed in 150 mg/g VS dosage of Ce-MOF (Ce-MOF-150) could be attributed to the enhanced sludge hydrolysis, acidogenesis, and methanogenesis processes, proven by the increased relative abundance of dominant acidogens and methanogens. Furthermore, 150 mg/g VS hydrolytic nanozyme bolstered the relative abundance of genes encoding acetyl-CoA carboxylase (accABCD) and involving in glycolysis (hxk and pfkA). These quantities were higher (8.31 − 34.14%) than that without nanozyme addition. The enhanced relative abundance of ackA, pta, cdhCDE, mvhAD, fpoABCDHIJKLMNO, vhoC, rnfABCDEG, mtrABCDEFGH, and hdrABC in the Ce-MOF-150 test indicated that the high proton conductivity of nanozyme facilitated acetoclastic methanogenesis and membrane-bound electron transport system, thereby enhancing methane production. These findings contribute to the expanding understanding of the application of artificial nanozymes as biocatalysts for solid waste resource recovery, providing a promising approach for methane production from sewage sludge.

Impact of food waste addition in energy efficient municipal wastewater treatment by aerobic granular sludge process

Recently, one of the main purposes of wastewater treatment plants is to achieve a neutral or positive energy balance while meeting the discharge criteria. Aerobic granular sludge (AGS) technology is a promising technology that has low energy and footprint requirements as well as high treatment performance. The effect of co-treatment of municipal wastewater and food waste (FW) on the treatment performance, granule morphology, and settling behavior of the granules was investigated in the study. A biochemical methane potential (BMP) test was also performed to assess the methane potential of mono- and co-digestion of the excess sludge from the AGS process. The addition of FW into wastewater enhanced the nutrient treatment efficiency in the AGS process. BMP of the excess sludge from the AGS process fed with the mixture of wastewater and FW (195 ± 17 mL CH4/g VS) was slightly higher than BMP of excess sludge from the AGS process fed with solely wastewater (173 ± 16 mL CH4/g VS). The highest methane yield was observed for co-digestion of excess sludge from the AGS process and FW, which was 312 ± 8 mL CH4/g VS. Integration of FW as a co-substrate in the AGS process would potentially enhance energy recovery and the quality of effluent in municipal wastewater treatment.Graphical abstract

Establishment of an inspection standard for decomposition and methane production rates of organic waste co-digestion facilities in Korea

Domestic organic waste resources have increased over the past decade and treatment of this waste via co-digested biogasification facilities is increasing annually. However, inspection standards for such facilities are not well-established. Herein, we aimed to derive calculation formulas and factors related to organic matter decomposition efficiency and methane production rate in accordance with waste treatment facility inspection standards. We also aimed to determine the optimum waste mixing ratio. Sample (field) surveys of 18 treatment facilities and complete enumeration of 110 facilities were conducted. Calculation formulas and factors were derived using the survey data and biochemical methane potential (BMP) test. The calculated coefficients derived through the BMP test were 0.512 m3 CH4/kgVSin for food waste, 0.601 m3 CH4/kgVSin for livestock manure, and 0.382 m3 CH4/kgVSin for sewage sludge. The final derived calculation factors were 65.0% for food waste, 36.0% for livestock manure, and 20.0% for sewage sludge for organic matter decomposition efficiency, and 0.380 m3 CH4/kgVSin for food waste, 0.27 m3 CH4/kgVSin for livestock manure, and 0.140 m3 CH4/kgVSin for sewage sludge for methane production rates. The derived effective capacity calculation factors can be utilized in future waste treatment facility inspection methods by aiding in the establishment of appropriate inspection standards for co-digested biogasification facilities other than single food waste treatment facilities. In addition, the optimum mixing ratio can be used as design data for co-digested biogasification facilities.

Anaerobic biodegradation of brown grease and its potential as a source of renewable energy

The purpose of the study was twofold. The first was to study the anaerobic digestibility and the biochemical methane potential (BMP) of brown grease (BG)–fats and oils collected from restaurants and similar food production facilities. The second objective was to investigate the effects of adding dry, food waste derived hydrochar to enhance the digestion process, specifically as a possible in situ agent to improve biogas quality. The BMP tests involved various BG loadings between 3 and 30 g BG/L. The results indicate that BG is highly digestible under anaerobic conditions with 354 mL CH4/g COD equivalent of BG at 1 atm and 35°C testing conditions, which translates into 28 million gasoline gallon equivalent (GGE) potential energy that could be recovered by anaerobically digesting or co-digesting BG. The particular hydrochar investigated in this study did not show any potential to increase biogas CH4 content.

Methane Production from Poly(lactic Acid) (PLA) and Polyhydroxybutyrate (PHB) Biodegradable Plastics with Anaerobic Granulated Sludge

- In this study, using Methanosarcina barkeri DSM 800 and Methanococcus vannielii DSM 1224 methanogenic bacteria under mesophilic (38 ± 1oC) and thermophilic (58 ± 1oC) conditions in anaerobic granulated sludge taken from Pakmaya Yeast Factory in Izmir, Turkey; Methane production from biodegradable plastics with poly(lactic acid) (PLA) and polyhydroxybutyrate (PHB) was investigated. Effect of different operating parameters, increasing biodegradation times (from 10 days to 500 days), different inoculumsubstrate ratios (ISRs) (16, 8, 4, 2, 1) and increasing biochemical methane potential (BMP) times (between 10 day and 500 days) for the production of methane gas from PLA and PHB biodegradable plastics in anaerobic granular sludge waste; Methanosarcina barkeri DSM 800 and Methanococcus vannielii DSM 1224 methanogenic bacteria were operated during the anaerobic digestion process under anaerobic conditions at mesophilic (38 ± 1oC) and thermophilic (58 ± 1oC) experimental temperatures. PLA biodegradable plastics were operated at optimum pH=7.6. PHB biodegradable plastics were carried out at optimum pH=8.1. Predicting the biodegradation behavior of PLA and PHB biodegradable plastics with BMP tests; It is found that the ISR parameter plays a very important role. This study showed that temperature plays a key role in the aging of microorganisms (Methanosarcina barkeri DSM 800 and Methanococcus vannielii DSM 1224 methanogenic bacteria) during anaerobic digestion, the degradation of bioplastic materials (PLA and PHB) and the degradation of their natural properties. The increase in temperature from mesophilic conditions to thermophilic conditions increased the activities of methanogenic bacteria such as Methanosarcina barkeri DSM 800 and Methanococcus vannielii DSM 1224. The maximum cumulative CH4(g) production was measured at 630 NL CH4 / kgVS for PHB biodegradable plastics in anaerobic granulated sludge with inoculum culture (the mixture of Methanosarcina barkeri DSM 800 and Methanococcus vannielii DSM 1224 methanogenic bacteria), at ISR=16 value, after 100 days, at pH=8.1 and at 58±1oC, respectively. The maximum 97% biodegradation efficiency was observed for PHB biodegradable plastics after 100 days, at pH=8.1 and at 58 ± 1oC thermophilic conditions, respectively.

Correction to: Alkaline pretreatment of tannery wastewater impact on biochemical methane potential tests: experimental study and kinetic modeling

Correction to: Alkaline pretreatment of tannery wastewater impact on biochemical methane potential tests: experimental study and kinetic modeling

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.

Biogas and Biohythane Production from Anaerobic Co-digestion of Canned Sardine Wastewater with Glycerol Waste

A biochemical methane potential (BMP) test investigated the effect of glycerol waste (GW) concentration on anaerobic co-digestion with canned sardine wastewater (CSW). They were studied using the single-stage process at mesophilic (P1) and thermophilic (P2) conditions and two-stage mesophilic (P3) processes. The P3 process has provided the most significant potential for improving biogas production in the sardine canning industry. Using 4% GW (v/v), the optimal hydrogen and methane concentrations at P3 are 43.00 ml H2/g CODr and 303.69 ml CH4/g CODr, respectively. The P3 process was 11.33 m3 biohytane/m3 mixed substrate, and the biohytane composition contained 43.11% CH4, 21.45% H2, and 35.43% CO2. The modified Gompertz model could simulate satisfactory hydrogen and methane yields, corresponding to high regression coefficients (R2>0.90). Hydrogen-producing bacteria in the H2 batch reactor were dominated by Micrococcus sp. and Desulfovibrio sp., while Methanosaeta sp., Methanoculleus sp., and Methanosarcina sp. are the major methanogens in the CH4 batch reactor. A two-stage process of co-fermenting CSW and GW could be a potential option for simultaneous biofuel recovery and waste treatment.

A method for evaluating and verifying biochemical methane potential test completion performed with landfilled municipal solid waste.

The biochemical methane potential (BMP) test is significant for the landfill industry as it provides a means to evaluate the gas potential, and therefore potential degradability, of both incoming and in-place municipal solid waste (MSW). However, the BMP test is not standardized making comparison of BMP results across sites problematic. For example, the BMP test duration has historically ranged from 20 days to several months with most current BMP tests lasting 60 days. However, the gas generation data can potentially be modelled for any of those durations to produce a prediction of the ultimate BMP value (BMPULT). Currently, the predicted BMPULT values of 23 long-duration (115-150 days) BMP tests were used to determine the required quantity of data (i.e. number of days) needed to produce an accurate BMPULT prediction. Results showed that no single test duration produced both accurate and efficient results, so a novel performance-based endpoint was proposed. The relative change in predicted BMPULT values with respect to time (dBMPULT/dt) was chosen as a potential performance-based completion metric. Results indicate that once the absolute normalized dBMPULT/dt value is within <2.5, <1.5 and <0.6% day-1 that the predicted BMPULT is within 20, 10 and 5% of the true BMPULT, respectively. Overall, the use of performance-based metrics for determining BMP test completion will allow for the collection of partial data sets, reduced experimental times and verification of results.

Effect of direct enzyme addition on the anaerobic digestion of OFMSW

The organic fraction of municipal solid waste (OFMSW) has high biodegradability and is a good feedstock for anaerobic digestion (AD). However, the solubilization of OFMSW is a limiting factor for AD. In this study, enzyme treatment was investigated to increase the conversion of complex materials of OFMSW into soluble fractions. Six different concentrations (0.15, 0.3, 0.5, 1.0, 1.5 and 3.0 mg/VSadded) of the alpha amylase enzyme (obtained from Aspergillus oryzae) were directly added as a powder and automated biochemical methane potential (BMP) tests were performed under thermophilic conditions. According to BMP tests, the methane (CH4) yield increased up to 17.5 % and the lag phase decreased to 19.3 % after enzyme addition. The highest CH4 yield and shortest lag phase were obtained when 0.5 mg of enzyme/VSadded was added to the reactor. These findings suggest that direct alpha amylase enzyme application obtained from Aspergillus oryzae can be used to increase biogas production by enhancing solubilization of OFMSW for AD.

Near-infrared reflectance spectroscopy for rapid prediction of biochemical methane potential of wastewater wasted sludge

The information of biochemical methane potential (BMP) of wasted sludge is essential to ensure the stable operation of sludge management processes. However, conventional anaerobic digestion (AD) approach for BMP test is time-consuming and labour-intensive. Currently, the technique of Near Infrared Spectroscopy (NIRS) is gaining prominence in the biogas production within AD process. Previous studies mostly focused on predicting BMP values for fibrous plant biomass and solid waste, with only a limited number of studies attempting to apply NIRS to obtain BMP values across a wide array of wasted sludge types. To obtain BMP values for this diverse range of wasted sludge efficiently and accurately, it is imperative to develop precise models for assessing BMP values using NIRS. In this study, the possibility of using NIRS to predict the BMP values of wasted sludge was evaluated. A total of 70 sludge samples from different sources were investigated to develop a BMP-prediction model by correlating the measured BMP values with the obtained NIR spectra. As a result, a reliable and successful BMP-prediction model was established with the determination coefficient of 0.90, residual prediction deviation of 3.50 and low root mean square error of prediction of 36.8 mL CH4/g VS. This BMP-prediction model is satisfactory for predicting BMP values of various types of sludge. It could provide support for plant operators to make decisions rapidly, thereby improving the process efficiency and optimizing sludge management procedures.

Assessment of biochemical methane potential of dairy wastewater with different co-substrates and evaluation of different kinetic models.

Dairy industry wastewater can be considered as an important source of pollution due to its high amounts and pollutant concentrations. Anaerobic treatment is seen as a suitable alternative over aerobic treatment which requires huge aeration systems. Biochemical methane potential (BMP) testing is a widely applied technique for estimating the performance of anaerobic digesters and still has no clear alternative. In the study, the biochemical methane potential change was investigated by mixing dairy wastewater with different co-substrates (cattle manure, chicken manure and slaughterhouse wastewater) at different rates. The highest biogas potential per gram of chemical oxygen demand added (CODadded) was determined as 574 mLbiogas in a mixture of 74% dairy wastewater + 2% chicken manure + 24% slaughterhouse wastewater inoculated with granular sludge. The highest methane potential was determined as 340 mLCH4 in the same co-substrate mixture inoculated with anaerobic sludge. In recent years, mathematical modeling offers an alternative to BMP tests and many different models are used for this purpose. In the study, six different mathematical models were used to simulate the BMP results, and the highest correlation coefficient in almost all mixtures ranged from 0.900 to 0.997 with the Modified Gompertz equation and Fitzhugh models.

Characterization and Methanogenic Potential Evaluation of Faecal Sludge: Case of the Kossodo Biogas Plant in Ouagadougou

The use of faecal sludge (FS) in anaerobic digestion (AD) requires a perfect knowledge of their composition. Considered as a very heterogeneous material, the high variability of FS can disturb biodigesters’ functionality and impact biogas production. Unique in West Africa, Kossodo’s biogas plant in Ouagadougou receives sludge from septic tanks and pit latrines. To evaluate the quality of sludge discharged in this treatment plant and its ability for AD, a characterization of 130 FS trucks from several onsite sanitation facilities was carried out. Physico-chemicals, including heavy metals and microbiological parameters, were analyzed using standard protocols. A biochemical methane potential test was employed to evaluate biogas yield. Results showed that raw sludge averaged 1.12% total solids (TS), 54.74% volatile solids (VS), 9253 mg/L chemical oxygen demand (COD), and 1645 mg/L biochemical oxygen demand (BOD). Settled faecal sludge exhibited higher levels of total coliforms, E. coli, helminth eggs, and heavy metals. Heavy metal levels met AD standards defined by VDI 4630, with decreasing toxicity order: Zn &gt; Mn &gt; Cu &gt; Cr &gt; Ni &gt; Pb &gt; As ≥ Hg. The carbon-to-nitrogen (C/N) ratio was 6.7 ± 4.3, indicating unsuitability for AD. Sludge settling increased C/N ratio by 46%, which was still below optimal AD conditions (20–30). Methane yield of raw and settled FS averaged 61 ± 0.2 and 156 ± 3.2 NL CH4/kg VS removed, respectively. Co-substrate addition could enhance the methanogenic yield of these sludges. This study provides a valuable database on the characteristics of FS, supporting sustainable recovery options.

Impact of dewatering inorganic coagulants on anaerobic digestion treating food waste leachate

Anaerobic digestion of food waste leachate (FWL) provides a viable solution for waste treatment and energy production. Returning solids from digested sludge to the reactor maintains a high microbial concentration and enhances digestion efficiency. However, this requires coagulants because the digestate has low dewaterability. This study analyzed methane production and microbial communities using biochemical methane potential tests for inorganic coagulants (AlCl3, Al2(SO4)3, FeCl3, and Fe2(SO4)3) in FWL treatment. Cumulative methane production was the highest in the control and decreased in the order of Fe2(SO4)3, AlCl3, FeCl3, and Al2(SO4)3. Iron ions inhibited H2S production while aluminum ions increased it compared to the control group. Despite the absence of significant changes in microbial communities following coagulant injection, a substantial increase in damaged cells was observed. These findings highlight the intricate repercussions of coagulant introduction in anaerobic digestion, emphasizing notable alterations in methane production dynamics and the integrity of microbial cells.

Chemically activated hydrochars as catalysts for the treatment of HTC liquor by catalytic wet air oxidation

Hydrothermal carbonization (HTC) is a highly efficient and valuable technology for treating wet solid wastes and producing solid carbon-based materials named hydrochar. In this work, a hydrochar coming from the HTC of an anaerobic digestion sludge of wastewater treatment plant was used to assess the influence of several activation agents, a base (KOH) and different chloride salts (FeCl3, ZnCl2, and CuCl2) with the exact molar quantities, to develop materials with enhanced surface area and potential inclusion of metal active species for application in wet air oxidation processes. The KOH as an activating agent increased the surface area of hydrochar up to ca. 1000 m2/g of BET surface area. The employment of CuCl2 and FeCl3 as activating agents allows Cu- and Fe-rich doped materials of remarkable surface areas with 49.1 and 42.5 wt% of each metal, respectively. Likewise, the catalytic behavior of the different synthesized carbon-based materials as metal-free and metal-doped catalysts was evaluated for the Catalytic Wet Air Oxidation (CWAO) of a HTC aqueous liquor from a HTC process of animal manure to produce a valuable stream of higher biochemical methane potential in anaerobic digestion. CWAO effluents increased the proportion of carboxylic acids as final by-products due to the oxidation of more complex organic compounds of the initial effluent (ketones, phenols, aromatics and olefins). The CWAO treatments improve the anaerobic digestion rate in biochemical methane potential tests, although the methane production was limited by the lower TOC concentration of the treated streams after CWAO. This research contributes to developing sustainable and efficient strategies for the HTC-liquor treatment, using its solid hydrochar as catalysts, closing the loop of a Circular Economy.

Evaluation of batch mesophilic anaerobic digestion of waste Euro banknotes for methane Production: Preliminary studies and kinetic approach

The European Central Bank is striving to find environmentally friendly alternative methods of waste disposal. In 2020, it decided to end the disposal of Waste Euro Banknotes (W€B) in landfills and planned to use them for recycling and/or energy recovery. Despite being recognized as an effective tool in a circular economy model, there are no reported studies on the use of W€B as a substrate in anaerobic digestion (AD). Thus, the aim of this research was to assess the viability of W€B to be converted into high-value-added energy products (mainly methane) through AD. W€B (€10 and €20 denominations) provided by the Bank of Spain were used. Biochemical methane potential (BMP) tests of pre-treated (grinded) and untreated W€B were assessed at mesophilic temperature. The ultimate methane yield was considerably higher for pre-treated W€B (334 ± 23 NmL CH4 g VS-1added) than for untreated W€B (297 ± 14 NmL CH4 g VS-1added). The Logistic or Sigmoidal kinetic model adequately fit the experimental data and allowed for obtaining the kinetic parameters of each case studied. In this sense, an increase of 22.4 % in the maximum methane production rate (Rmax) was observed for the pre-treated W€B (52.5 ± 0.9 mL CH4 (g VS·d)-1 compared to the untreated W€B (16.2 ± 1.8 mL CH4 (g VS·d)-1). According to the obtained results, AD may be a good alternative for the energetic valorization and recycling of W€B.

Machine learning-based prediction of methane production from lignocellulosic wastes

The biochemical methane potential test is a standard method to determine the biodegradability of lignocellulosic wastes (LWs) during anaerobic digestion (AD) with disadvantages of long experiment duration and high operating expense. This paper developed a machine learning model to predict the cumulative methane yield (CMY) using the data of 157 LWs regarding physicochemical characteristics, digestion condition and methane yield, with the coefficient of determination equal to 0.869. Model interpretability analyses underscored lignin content, organic loading, and nitrogen content as pivotal attributes for CMY prediction. For the feedstocks with a cellulose content exceeding about 50%, the CMY in the early AD stage would be relatively lower than those with low cellulose content, but prolonging digestion time could promote methane production. Besides, lignin content in feedstock surpassing 15% would significantly inhibit methane production. This work contributes to valuable guidance for feedstock selection and operation optimization for AD plants.

Evaluating Biogas Production of Polylactic acid with the addition of Conductive Materials

Objectives:The demand for polylactic acid (PLA) is increasing as an alternative to conventional petroleum-based plastics due to its eco-friendly characteristics. PLA can be processed through anaerobic digestion (AD) along with other organic wastes to promote efficient and controlled biodegradation. This study aimed to evaluate the feasibility of biogas production through AD of PLA and to enhance efficiency through the addition of conductive materials.Methods:Two sets of biochemical methane potential (BMP) tests were conducted. The first BMP test involved substrate/inoculum ratios (SIRs) of 0.6, 0.8, 1.0, 1.2, 1.4, and 1.6 (g COD PLA/g COD seed sludge). Conductive materials powdered activated carbon (PAC), carbon nanotube (CNT), and Magnetite were individually added to reactors at a concentration of 3 g/L in the second test.Results and Discussion:The study revealed an increasing trend in methane production and methane yield with higher levels of PLA input. However, a decline in methane yield was observed when PLA was injected at levels surpassing 35g COD/L. This phenomenon can be attributed to a prolonged Lag phase, indicating a longer adaptation period for microorganisms, consequently resulting in a reduction of the conversion rate from 1g COD substrate to methane. The introduction of conductive material led to elevated cumulative methane production and methane yield in comparison to the control group. Notably, Magnetite exhibited the highest increase rate among the tested materials. Additionally, the addition of PAC demonstrated favorable results in terms of methane production rate at 99.4mL/L/Day and a Lag phase of 5.7 days. Conclusion:Experiments were conducted to evaluate the effects of PLA biogas production and the injection of PAC, CNT, and Magnetite at various substrate/inoculum ratios. When PLA was used as a substrate for anaerobic digestion, its feasibility was confirmed. However, it exhibited lower efficiency compared to other substrates. Therefore, the addition of a conductive material was found to increase the biogas production efficiency.

ENHANCEMENT OF BIOGAS GENERATION BY UTILIZING RAW AND MODIFIED WITH HNO3 BIOCHAR OBTAINED FROM PYROLYSIS OF BIOMASS AND DIGESTATE

Biomass- and digestate-derived biochars were modified with nitric acid solution and examined in biochemical methane potential (BMP) tests to determine the effect of pretreatment on each of the different materials capability to improve the biogas production from the anaerobic digestion of conventional substrates such as corn silage. Methane yields from corn silage (0.308 m3kgVS-1) were over the average value for the specific type of lignocellulosic material. Addition of digestate-derived biochar (BCD) in AD process improved the methane production 1.13-fold. However, the sawdust-derived biochar (BCS) resulted in an even greater methane release of 0.374 m3kgVS-1. Chemical treatment reduced the pH of BCs from 10.29 and 11.54 to 3.10 and 2.81 for BCS and BCD, respectively while had a significant impact on materials composition almost removing the ash and metal elements from BCS and markedly decreasing 1.43-fold the ash fraction and by 70-75 % the minerals proportion in BCD. The presence of modified digestate-derived biochar (M-BCD) in a culture led to an enhanced methane production (0.402 m3kgVS-1) indicating that the specific additive exhibited a higher potential than all BCs to promote the efficiency of AD of a biomass feedstock. M-BCD also possessed the greatest capability to lessen an inhibition caused by H2S retaining the concentration of the toxic gas at levels lower than 100 ppm. On the other hand, modified BCS provoked a 9% abatement in methane yields providing evidence that nitric acid could have a neutral or slightly negative effect on the capability of a BC to improve the AD process.