Search Results (20,463)

Water contamination with surface-active agents, such as sodium lauryl sulfate (SLS), represents a serious environmental concern, driving the need for efficient and low-cost alternative adsorbents as a step toward sustainable waste valorization. In this study, waste biomass derived from plum stones, date stones, and walnut shells was successfully transformed into activated carbons via chemical activation using potassium hydroxide (KOH) at 850 °C with a 1:1 impregnation ratio. The synthesized materials underwent comprehensive physicochemical characterization utilizing TG-DSC, elemental analysis, Boehm titration, SEM, TEM, and nitrogen physisorption (BET), whereas their adsorption performance was evaluated against aqueous SLS solutions across various concentrations. The obtained results reveal a predominantly microporous structure with a high specific surface area, reaching up to 1059.01 m²/g for ACdate. The equilibrium adsorption data were well described by the Langmuir isotherm model, which yielded model-estimated asymptotic adsorption capacities (q_m) of 219.70 mg/g for ACwalnut, 178.25 mg/g for ACdate, and 57.80 mg/g for ACplum. These values represent Langmuir-derived model parameters rather than experimentally attained adsorption capacities within the investigated concentration range. Notably, despite having a lower specific surface area than ACdate, ACwalnut exhibited the highest Langmuir-estimated q_m, which may be associated with its structural balance and well-developed mesoporous network (0.210 cm³/g), facilitating the intraparticle transport of SLS molecules. These findings highlight that high efficiency originates from a synergistic combination of accessible porosity, a mesoporous transport network, hydrophobic character, and specific surface functional groups, demonstrating the exceptional potential of these activated carbons for anionic surfactant wastewater remediation. Full article

Poly(β-L-malic acid) (PMLA) is a water-soluble, biodegradable aliphatic polyester whose pendant carboxyl groups support chemical functionalization for biomedical, packaging, and materials applications. Microbial fermentation can use pure sugars and biomass-derived carbon sources under mild conditions, but industrial translation remains constrained by feedstock cost and variability, strain performance, oxygen and pH control, pretreatment-derived inhibitors, and downstream recovery. This review therefore focuses on the fermentative production of PMLA from refined and renewable carbon sources, the microorganisms and metabolic routes involved, and the process variables that govern titer, yield, productivity, molecular weight, and purification. Agricultural and forestry feedstocks are compared according to their actual carbohydrate class and processing requirements. Bamboo shoot shell hydrolysate is treated as an emerging case study rather than an established production platform: one accepted shake-flask study directly demonstrated PMLA production by Aureobasidium pullulans NRRL Y-2311-1, but controlled bioreactor validation, reproducibility, techno-economic analysis, and application-specific product qualification remain to be further investigated. The review also examines autohydrolysis, low-molecular-weight PMLA for biomedical use, furan inhibition, membrane and ion-exchange purification, and the limits of current economic comparisons. This evidence-based framing identifies where bamboo-processing residues may contribute to renewable PMLA production while distinguishing laboratory feasibility from industrial readiness. Full article

Gansu Province has abundant agricultural biomass resources, but the gap between the biological yield of biomass and available quantities, along with strong spatial heterogeneity, restricts efficient bioenergy utilization. This study quantifies four categories of agricultural biomass across 85 county-level units in Gansu, evaluates relative enrichment density via the Biomass Resource Location Quotient (BRLQ), and identifies spatial agglomeration patterns using global and local spatial autocorrelation. The results show a significant structural mismatch: total biological yield of biomass reaches 46.41 million tons, while available resources are only 12.13 million tons, with crop straw as the dominant available feedstock. The Longdong Loess Plateau has the highest enrichment density (BRLQ = 1.588). No significant global spatial agglomeration is observed, and significant clusters are confined to local small areas. This study provides quantitative support for zonal bioenergy planning in Gansu. Full article

In silico studies of microbiological systems are essential for predicting and controlling the impact of external factors on bacterial communities. Quorum sensing represents one of the key mechanisms of bacterial communication, particularly in pathogenic bacteria, realized as a cell-density-dependent regulatory process governed by diffusible signaling molecules. The present study proposes a Physics-Informed Neural Network (PINN)-based computational framework for a spatially independent model of bacterial quorum sensing and population dynamics. The approach solves both the forward and inverse problems for a spatially independent model formalized by a system of nonlinear ordinary differential equations. The forward problem is solved numerically by reconstructing the dynamics of three key characteristics: signaling molecule concentration, degrading enzyme concentration, and bacterial biomass density. The obtained PINN solutions are compared with numerical solutions computed using the Radau IIA implicit Runge–Kutta method. The inverse problem capability is evaluated by recovering system parameters that are difficult to measure directly in experimental settings. The framework is implemented using the DeepXDE library with a PyTorch backend, employing hard constraints for initial conditions, singularity-avoiding loss reformulations, and a multi-stage Adam–L-BFGS optimization strategy. Validation is performed on a Monod chemostat benchmark and the Pseudomonas putida IsoF quorum sensing regulatory network. The proposed PINN-based framework extends the applied mathematical toolkit for in silico studies of microbial systems, enabling accurate reconstruction of emergent population dynamics and robust inference of regulatory parameters that are inaccessible to direct experimental measurement. Full article

The application of biochar and bioorganic fertilizer (BOF) in agricultural systems has garnered increasing attention in recent years. Nevertheless, research remains scarce on the impacts of biochar and BOF on the rhizosphere microecological characteristics of saline-alkali soils. This research involved the execution and analysis of 16S rRNA sequencing using Illumina technology to explore how biochar, whether used alone or in conjunction with BOF, along with varying application rates, impacts the microbial community in the saline-alkali rhizosphere soil during quinoa cultivation. In the conducted field trial, sole BOF application, sole biochar application, and their combined application (referred to as BOFB) led to a substantial enhancement of 23.88%, 74.08–97.00%, and 188.88–220.59% in quinoa aerial biomass, respectively. Meanwhile, sole biochar application or biochar combined with BOF reduced soil electrical conductivity (EC) by 26.42–39.81%. Biochar and BOF significantly improved most soil parameters, with the exception of total phosphorus (TP). In comparison to the control (CK), the relative abundances of Pseudomonas, Arthrobacter, Skermanella, and Bacillus were elevated in the biochar and BOFB treatments, while Sphingomonas was more abundant in the BOF treatment. In addition, Skermanella exhibited a significant positive correlation with EC and available potassium (AK). Biochar exerted a stronger effect on soil bacterial community structure than BOF. Furthermore, the complexity of the bacterial community in biochar and BOFB treatments far exceeded that in the BOF and CK treatments. Overall, the application of biochar effectively reduced soil EC and improved soil fertility, enhanced bacterial community stability, and optimized bacterial community structure, thereby increasing quinoa aerial biomass. Under the conditions of this study, the optimal application rate for biochar was 15 t/ha, and the combined application of biochar and BOF produced superior effects relative to either amendment alone. Full article

Brackish water utilization provides an alternative strategy for alleviating freshwater scarcity in arid agricultural regions; however, the synergistic regulation of salinity, irrigation, and nitrogen management remains unclear. A two-year field experiment was conducted in 2025 and 2026 to investigate the effects of water–nitrogen–salinity interactions on growth, yield formation, resource use efficiency, and fruit quality of pigment pepper (Capsicum annuum L.) under arid conditions in Xinjiang, China. An L9(3³) orthogonal experimental design was adopted with three levels of brackish water salinity, irrigation amount, and nitrogen application rate. The comprehensive production performance of different management strategies was further evaluated using a combined weighting Cloud–TOPSIS approach. The results showed that water–nitrogen–salinity interactions significantly regulated pigment pepper growth, yield formation, and resource utilization, with consistent responses observed across the two experimental years. Increasing irrigation water salinity reduced leaf chlorophyll content (CHL) and nitrogen balance index (NBI), whereas flavonoid content (FLAV) exhibited an increasing trend under moderate salinity stress. Low-salinity irrigation combined with appropriate water and nitrogen inputs maintained higher photosynthetic capacity and nitrogen nutritional status. Yield, water use efficiency (WUE), and partial factor productivity of nitrogen (PFPN) were jointly affected by salinity, irrigation, and nitrogen supply. Excessive salinity significantly reduced crop productivity, while optimized irrigation and nitrogen management alleviated salt stress effects. The T2 treatment (1 g L⁻¹ salinity, 2400 m³ ha⁻¹ irrigation, and 300 kg ha⁻¹ nitrogen application) achieved the highest yield and maintained favorable WUE and PFPN values in both years. Fruit quality responses demonstrated that moderate salinity promoted capsaicinoid accumulation, whereas excessive salinity restricted biomass production and quality improvement. Correlation analysis revealed that photosynthetic nitrogen metabolism indicators were closely associated with yield formation, while flavonoid accumulation showed stronger relationships with quality attributes. The Cloud–TOPSIS evaluation identified T2 as the optimal management strategy under the experimental conditions by balancing yield, quality, and resource use efficiency. These findings indicate that coordinated regulation of irrigation water salinity, water supply, and nitrogen input is essential for achieving efficient brackish water utilization and sustainable pigment pepper production in arid regions. Full article

In alkaline soils rich in residual phosphorus, phosphorus availability to plants depends not only on the existing reserve, but also on the redistribution of the extractable fraction within the root-explored zone. This study evaluated the effects of water regime, fertilizer treatment, and wheat presence on the distribution of ammonium lactate-extractable phosphorus (P-AL) within the 2–8 cm layer of an alkaline calcareous Chernozem with high initial P availability (P-AL = 178.1 mg kg⁻¹). The pot experiment compared plant-free soil (S0) and wheat-planted soil (SP), four water regimes (H0–H150; 0–150 L m⁻²), and four fertilization treatments: an unfertilized control (F0), a solid NPK fertilizer (FS), and a liquid NP fertilizer applied at low and high rates (FL1 and FL2). These treatments represented practical fertilization options and were not equivalent in P input, supplying 51.8, 13.9, and 27.9 mg P pot⁻¹ for FS, FL1, and FL2, respectively. Soil and plant samples were collected at BBCH 21–22, 20 days after fertilization. Water regime was a major factor shaping P-AL redistribution, significantly affecting P-AL at all three analyzed soil depths (p < 0.001), with its effect depending on vegetation condition and fertilization treatment. Wheat presence reduced P-AL relative to S0, and apparent P-AL depletion (ΔP-AL = S0 − SP) was greatest under H0, ranging from 83 to 99 mg kg⁻¹. FL2 produced the largest S0–SP contrasts under H0–H100, whereas under H150 the largest difference was associated with FS. Under high water input, the higher-input solid NPK treatment (FS), which supplied the largest P input and was the only treatment supplying K, was associated with the highest shoot biomass (16.7 g), root biomass (6.92 g), and root P accumulation (25.2 mg pot⁻¹). The results indicate that P fertilization in high-P alkaline soils should be adapted to water regime, fertilizer input and application method, without allowing for direct conclusions regarding phosphorus use efficiency, total plant P uptake, or leaching losses. Full article

The development of sustainable, biocompatible scaffolds with enhanced structural stability remains a primary challenge in bone tissue engineering. In this study, structurally reinforced nanocomposite scaffolds were successfully fabricated by integrating green-synthesized carbon quantum dots (CQDs) into a gelatin (G) matrix, offering an innovative platform that mimics the organic–inorganic interfaces of natural bone tissue. The CQDs were derived from oatmeal via a sustainable, green hydrothermal route, serving simultaneously as zero-dimensional reinforcing fillers and bioactive agents within the biopolymer network. To ensure an additive-free fabrication process that avoids toxic chemical cross-linkers, dehydrothermal (DHT) treatment was employed, successfully modulating the interfacial and chemical cross-linking interactions between the gelatin chains and the oxygen-rich surface groups of the CQDs. Structural characterization confirmed the uniform dispersion of CQDs (average diameter 7–8 nm) within the porous gelatin framework. The incorporation of CQDs significantly improved the physicochemical properties of the scaffolds; the G/CQD 5% formulation emerged as the optimal composition, exhibiting a 118% increase in compression modulus compared to pristine gelatin. The composite demonstrated tuned swelling kinetics and a significantly reduced degradation rate, restricting mass loss after 14 days of incubation to approximately 24% compared to 40% for pristine gelatin, which is essential for maintaining a structural template during the initial stages of tissue formation. Bioactivity assays in simulated body fluid (SBF) confirmed the rapid, biomimetic induction of a crystalline hydroxyapatite layer with a natural Ca/P ratio of 1.61 within 14 days. Furthermore, preliminary in vitro assessments using Human Skin Fibroblasts (HSFs) confirmed excellent general cytocompatibility, with cell viability exceeding 90%. This study highlights the unique potential of utilizing biomass-derived carbon nanostructures and clean manufacturing processing to engineer multifunctional scaffolds with enhanced structural stability and intrinsic bioactivity for potential bone defect repairs. Full article

Cibotium barometz (L.) J.Sm., a medicinal fern, relies on three interconnected processes for in vitro propagation: spore germination, ordered gametophyte development, and successful initiation and early establishment of the sporophyte generation. However, during aseptic subculture, gametophytes frequently deviate from this developmental trajectory. The predominant abnormalities include filamentous arrest, thick fan-shaped blades, and irregular spatulate blades, all of which were associated with reduced archegonial maturation and subsequent sporophyte establishment. In this study, developmental time-course observation, paraffin-section histology, and medium-regulation experiments were integrated to establish a diagnosis–recovery–initiation framework for Cibotium barometz gametophytes. Spore germination and gametophyte development were divided into eight practical checkpoints: spore imbibition, spore-wall rupture, rhizoid emergence, rhizoid elongation, filamentous prothallus, lamellar prothallus, cordate gametophyte, and sporophyte formation. The ontogeny of archegonia was delineated into six histological stages, ranging from the initial-cell stage to the pre-fertilization stage. Based on an integrative assessment of population appearance, individual morphology, and anatomical sections, abnormal gametophytes were classified into three diagnostic categories. Filamentous arrest manifested as wool-like or thread-like growth patterns, featuring persistent chain-like cellular organization and failure to develop a two-dimensional blade. Thick fan-shaped blades showed pronounced enlargement and thickening; section images revealed narrow, densely packed cells, accumulation of storage compounds, and localized wall thickening, a pattern consistent with, but not sufficient to demonstrate, a preferential shift toward vegetative proliferation. Irregular spatulate blades were narrow and asymmetric and showed morphological and histological features suggestive of disrupted tissue polarity. Although archegonial initial cells or primordium-like structures were intermittently detected, they infrequently advanced to mature archegonia. Medium treatment significantly affected arrest rate, hypertrophy rate, sporophyte initiation, and mean young-sporophyte height (p < 0.001). Murashige and Skoog (MS) medium supplemented with IAA, 6-BA, and activated carbon was associated with the highest arrest rate, whereas full-strength MS medium was associated with the most pronounced hypertrophy. Among hormone-free subculture treatments, 1/2 MS + activated carbon gave the highest sporophyte initiation rate (71.45% ± 9.16%); the same formula also performed well in primary hormone-free cultures (70.54% ± 9.07%). Correlation analysis showed a strong positive association between sporophyte initiation rate and mean young-sporophyte height (r = 0.935, p < 0.001), while arrest and hypertrophy were negatively associated with sporophyte initiation. These results support evaluating in vitro propagation using quantified developmental outcomes—arrest rate, hypertrophy rate, sporophyte initiation rate, and mean young-sporophyte height—rather than biomass alone. Under the tested conditions, hormone-free 1/2 MS + activated carbon was associated with the most favorable recovery outcomes; the underlying mechanisms require validation by quantitative histology, gene-expression analysis, endogenous-hormone profiling, and direct measurement of compounds adsorbed by activated carbon. Full article

Reliable spatial evidence can support quality assurance and quality control for land use, land-use change and forestry (LULUCF), but land-cover and species-distribution layers do not by themselves identify IPCC Forest Land or species-pure stands. We estimated 2010 and 2020 above- and below-ground living biomass carbon within tree-covered European beech (Fagus sylvatica L.) distribution and occurrence footprints in Greece. Our operational hypothesis was that increasingly restrictive species masks would materially alter the mapped extent and carbon estimates. ESA Climate Change Initiative Biomass v6, ESA WorldCover 2021, European Forest Genetic Resources Programme (EUFORGEN) polygons, and Forest Information System for Europe (FISE) relative probability of presence layers were processed in Google Earth Engine. Biomass was converted with IPCC default carbon fractions and root:shoot ratios, and the results were summarized nationally and for GAUL Level-2 units. The broad EUFORGEN footprint covered 22,133 km², whereas the Combined overlap of EUFORGEN, FISE relative probability of presence ≥ 0.50, and tree cover covered 2742 km². Within the Combined footprint, the pixel mean living biomass carbon density was 60.33 Mg C ha⁻¹ in 2010 and 62.50 Mg C ha⁻¹ in 2020, and the area-integrated change was +0.58 Tg C; the area-normalized regional change was positive in 13 of 17 units and negative in 4. Across masks, the mean decadal change ranged from −0.50 to +3.37 Mg C ha⁻¹ and the approximate area-integrated totals from −1.10 to +0.58 Tg C. These scenario-conditioned estimates are neither official national greenhouse gas inventory estimates nor tests of statistical significance; instead, they provide reproducible spatial screening while making mask sensitivity and unpropagated uncertainty explicit. Full article

Implant-associated infections remain a major challenge in bone-related biomedical applications, where bacterial colonization and biofilm formation can compromise tissue integration and clinical performance. This study reports the mechanochemical synthesis, physicochemical characterization, antimicrobial activity, antibiofilm performance, and short-term fibroblast cytocompatibility of a TiO₂-containing bovine-derived biogenic hydroxyapatite ceramic composite (BHAp-TiO₂). The composite was prepared by high-energy mechanical milling using 10 wt% TiO₂ and characterized by X-ray diffraction, Rietveld refinement, Raman spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. XRD/Rietveld analysis identified a multiphase ceramic composite composed of hydroxyapatite, whitlockite, and rutile TiO₂, with no evidence of Ti⁴⁺ substitution into the hydroxyapatite lattice or detectable anatase within the XRD/Rietveld detection limit. SEM-EDS confirmed the granular agglomerated morphology of the powders and the elemental presence of Ti in BHAp-TiO₂. Compared with pristine BHAp, BHAp-TiO₂ produced a concentration-dependent reduction in AlamarBlue^®-derived bacterial metabolic activity against five clinically relevant planktonic strains. At 200 µg/mL, residual metabolic activity decreased to 10.90–32.90%, depending on the bacterial species, with the strongest response observed for Escherichia coli. In crystal violet assays, BHAp-TiO₂ markedly inhibited Pseudomonas aeruginosa biofilm biomass, reaching 91.9 ± 3.4% inhibition at 200 µg/mL. In NIH/3T3 fibroblasts, BHAp-TiO₂ preserved short-term cytocompatibility after 24 h of direct exposure within the 0.1–100 µg/mL range, with MTT- and AlamarBlue^®-derived responses remaining close to or above the 80% cytotoxicity limit. Overall, BHAp-TiO₂ is best interpreted as a rutile TiO₂-containing biogenic calcium phosphate ceramic composite with enhanced antimicrobial and antibiofilm performance while maintaining short-term fibroblast cytocompatibility under the evaluated conditions. Full article

The intrinsic weakness of the budding yeast Saccharomyces cerevisiae in xylose utilization limits its application in biological manufacturing using lignocellulosic biomass. Although the natural yeast strain S. cerevisiae YB-2625 exhibits superior innate xylose-fermenting capability, the underlying mechanisms remain largely unexplored. Here, we employed comparative multi-omics to systematically dissect the molecular basis of its high stress tolerance and superior xylose consumption. Comparative genomics revealed 73,842 single nucleotide polymorphisms (SNPs) and 5191 small insertions/deletions (InDels) in YB-2625 relative to S288C, with significant enrichment in genes associated with chromatin remodeling, transcriptional regulation, and stress signaling. Integration of genomic and transcriptomic data identified candidate variants in key regulators. Functional validation further demonstrated that Tra1, a component of the SAGA, SLIK, and NuA4 histone acetyltransferase complexes, acts as a global regulator with growth-coupled effects on stress tolerance and xylose metabolism. Deletion of TRA1 significantly reduced the final biomass in xylose medium. Moreover, deletion of RTT109 specifically impaired growth on xylose without affecting any of the tested stress tolerance phenotypes. We further examined global chromatin accessibility changes upon deletion of the histone acetyltransferase gene NGG1, a manipulation previously shown to substantially enhance xylose utilization in the engineered YB-2625 background. ATAC-seq analysis revealed that loss of Ngg1 alters chromatin accessibility at loci governing carbohydrate metabolism and stress responses, thereby establishing a direct link between epigenetic remodeling and the superior phenotype of YB-2625. Our findings provide a basis for deciphering the regulatory circuitry governing xylose utilization in recombinant yeast and for the rational engineering of robust strains for lignocellulosic bioconversion. Full article

Botrytis cinerea is a major fungal pathogen of grapevine responsible for gray mold, causing significant economic losses worldwide. Under specific environmental conditions, controlled development of Botrytis cinerea may lead to “noble rot,” a quality-enhancing attribute of premium wines; however, the same fungus can rapidly develop into destructive gray mold, causing significant grape losses. Consequently, the challenge is not necessarily the complete eradication of the pathogen, but the regulation of its development to prevent uncontrolled disease progression while preserving grape quality. The present study investigated the potential of using essential oils extracted from selected aromatic plants using ultrasound-assisted hydrodistillation (UAHD) as eco-friendly biocontrol agents against Botrytis cinerea. The extraction process was optimized by evaluating the effects of biomass particle size, ultrasonic power, and distillation time on essential oil yield. Antifungal activity was assessed through in vitro poisoned food and disk diffusion assays and validated in vivo on artificially inoculated grape berries by evaluating disease incidence, disease severity, treatment efficacy, and weight loss. Ultrasound pretreatment increased essential oil yield by up to 29.58% compared with conventional hydrodistillation, with optimal conditions achieved using 1 cm biomass, 150 W ultrasonic power, and 85 min distillation. Oregano, thyme, sage, and lemon balm essential oils achieved 100% inhibition of Botrytis cinerea mycelial growth at the tested concentrations, whereas lavender essential oil showed 95.03% inhibition at 0.125% and complete inhibition at 0.25%. In vivo, essential oil formulations markedly reduced gray mold development, with the 2.0% formulation providing the highest treatment efficacy and the lowest disease severity. These findings show that UAHD-derived essential oils represent promising sustainable alternatives for the management of Botrytis cinerea in viticulture. However, these findings represent preliminary evidence obtained under controlled in vitro conditions, necessitating further validation in real-world applications. Full article

The dynamics of aboveground biomass (AGB) in rubber plantations (RPs) provide an important basis for evaluating carbon stocks and environmental adaptability in tropical plantations. However, continuous monitoring of AGB of RPs at the regional scale is lacking, and its nonlinear responses to hydrothermal conditions remain insufficiently understood. This study focused on RPs in western Hainan Island (WHI), including Danzhou, Baisha, Lingao, and Chengmai, and integrated field plot data with multi-source remote sensing datasets. A framework for mapping RPs combining rule-based constraints and phenology-based random forest (RF) classification was developed. After key variable screening, extreme gradient boosting (XGBoost), Shapley additive explanations (SHAP), and generalized additive model (GAM) were used for AGB estimation and identification of climatic responses. The results showed that mapping of RPs achieved an overall accuracy of 92.89% and a Kappa coefficient of 0.854. The XGBoost-derived estimates showed that AGB of RPs in the study area increased by approximately 1.43 × 10⁶ Mg from 2017 to 2025, with growth areas mainly concentrated in the Danzhou–Baisha and western Chengmai. AGB exhibited significant nonlinear responses to climatic factors. Specifically, the effect of precipitation (PRE) shifted to negative after approximately 1945 mm yr⁻¹, whereas annual mean maximum temperature (TMAX) shifted to a positive effect after about 29.72 °C, although this effect gradually weakened as temperature continued to rise. Combinations such as PRE × annual mean temperature (PRE × TMP), PRE × TMAX, and PRE × potential evapotranspiration (PRE × PET) exhibited significant nonlinear interactions, indicating that the direction and magnitude of the effect of PRE shifted with changes in temperature and PET levels. These findings link the spatiotemporal changes in AGB of RPs in WHI with hydrothermal thresholds and their interacting effects, deepening our understanding of the climatic response characteristics of AGB in RPs in this region. They also provide a scientific basis for RP monitoring, carbon stock assessment, and climate-adaptive management in WHI. Full article

Conventional anaerobic digestion relies on diverse inocula present in sludge-based systems. A defined consortium approach was investigated as an alternative. Acetivibrio cellulolyticus was chosen as the cellulose degrader, and two strains of Methanosarcina barkeri were selected as methane producers. Initial cultivation following manufacturer protocols highlighted significant challenges in maintaining strict anaerobic conditions, particularly in the absence of specialized infrastructure. A simplified anaerobic cultivation workflow was therefore evaluated for pure cultures of the selected anaerobes and subsequently used to evaluate a defined consortium using microcrystalline cellulose (MCC) and industrial lignocellulosic biomass (LB) residue as growth substrates. The workflow enabled successful cultivation of pure cultures in their recommended nutrient media without detectable contamination, as assessed by microscopy, aerobic contamination checks, and gas chromatography analysis. Growth-associated observations were obtained on MCC after prolonged incubation (~30 days); however, a metabolically active cellulolytic–methanogenic consortium was not established, as methane was not detected and no activity was detected on the LB substrate. This study demonstrates that anaerobic cultivation of fastidious microorganisms is feasible using a simplified method without fully controlled anaerobic environments and highlights inherent challenges associated with the defined consortium on substrates such as MCC and complex LB residue. The simplified workflow may provide an accessible approach to anaerobic cultivation for sustainable biomethane research in laboratories lacking specialized anaerobic infrastructure. Further work is required to determine conditions supporting methane production using the defined consortium. Full article