The comprehensive effects of aluminum oxide nanoparticles on the physiology of freshwater microalga Scenedesmus obliquus and it's phycoremediation performance for the removal of sulfacetamide.

Nanoparticles are inevitable byproducts of modern industry. However, the environmental impacts arising from industrial applications of nanoparticles are largely under-reported. This study evaluated the ecotoxicological effects of aluminum oxide nanoparticles (Al2O3NP) and its influence on sulfacetamide (SA) biodegradation by a freshwater microalga, Scenedesmus obliquus. Although Al2O3NP showed limited toxicity effect on S. obliquus, we observed the toxicity attenuation aspect of Al2O3NP in a mixture of sulfacetamide on microalgae. The addition of 100 mg L-1 of Al2O3NP and 1 mg L-1 of SA reduced total chlorophyll by 23.3% and carotenoids by 21.6% in microalgal compared to control. The gene expression study demonstrated that ATPF0C, Lhcb1, HydA, and psbA genes responsible for ATP synthesis and the photosynthetic system were significantly downregulated, while the Tas gene, which plays a major role in biodegradation of organic xenobiotic chemicals, was significantly upregulated at 1 and 100 mg L-1 of Al2O3NP. The S. obliquus removed 16.8% of SA at 15 mg L-1 in 14 days. However, the removal was slightly enhanced (18.8%) at same concentration of SA in the presence of 50 mg L-1 Al2O3NP. This result proves the stability of sulfacetamide biodegradation capacity of S. obliquus in the presence of Al2O3NP co-contamination. The metabolic analysis showed that SA was degraded into simpler byproducts such as sulfacarbamide, sulfaguanidine, sulfanilamide, 4-(methyl sulfonyl)aniline, and N-hydroxy-benzenamine which have lower ecotoxicity than SA, demonstrating that the ecotoxicity of sulfacetamide has significantly decreased after the microalgal degradation, suggesting the environmental feasibility of microalgae-mediated wastewater technology. This study provides a deeper understanding of the impact of nanoparticles such as Al2O3NP on aquatic ecosystems.

Water condition in biotrickling filtration for the efficient removal of gaseous contaminants.

Biofiltration (BF) facilitates the removal of organic and inorganic compounds through microbial reactions. Water is one of the most important elements in biotrickling filters that provides moisture and nutrients to microbial biofilms. The maintenance of proper trickle watering is very critical in biotrickling filtration because the flow rate of the trickling water significantly influences contaminant removal, and its optimal control is associated with various physicochemical and biological mechanisms. The lack of water leads to the drying of the media, creating several issues, including the restricted absorption of hydrophilic contaminants and the inhibition of microbial activities, which ultimately deteriorates the overall contaminant removal efficiency (RE). Conversely, an excess of water limits the mass transfer of oxygen or hydrophobic gases. In-depth analysis is required to elucidate the role of trickle water in the overall performance of biotrickling filters. The processes involved in the treatment of various polluted gases under specific water conditions have been summarized in this study. Recent microscopic studies on biofilms were reviewed to explain the process by which water stress influences the biological mechanisms involved in the treatment of hydrophobic contaminated gases. In order to maintain an effective mass transfer, hydrodynamic and biofilm conditions, a coherent understanding of water stress and the development of extracellular polymeric substances (EPS) in biofilms is necessary. Future studies on the realistic local distribution of hydrodynamic patterns (trickle flow, water film thickness, and wet efficiency), integrated with biofilm distributions, should be conducted with respect to EPS development.

Efficient removal of formaldehyde using metal-biochar derived from acid mine drainage sludge and spent coffee waste.

A novel metal-biochar (Biochar/AMDS) composite were fabricated by co-pyrolysis of spent coffee waste (SCW)/acid mine drainage sludge (AMDS), and their effective application in adsorptive removal of air pollutants such as formaldehyde in indoor environments was evaluated. The physicochemical characteristics of Biochar/AMDS were analyzed using SEM/EDS, XRF, XRD, BET, and FTIR. The characterization results illustrated that Biochar/AMDS had the highly porous structure, carbonaceous layers, and heterogeneous Fe phases (hematite, metallic Fe, and magnetite). The fixed-bed column test showed that the removal of formaldehyde by Biochar/AMDS was 18.4-fold higher than that by metal-free biochar (i.e., SCW-derived biochar). Changing the ratio of AMDS from 1:6 to 1:1 significantly increased the adsorption capacity for formaldehyde from 1008 to 1811 mg/g. In addition, thermal treatment of used adsorbent at 100 °C effectively restored the adsorptive function exhausted during the column test. These results provide new insights into the fabrication of practical, low-cost and ecofriendly sorbent for formaldehyde.

Biodegradation of high concentration phenol using sugarcane bagasse immobilized Candida tropicalis PHB5 in a packed-bed column reactor.

Biodegradation of phenolic compounds in wastewater can be effectively carried out in packed bed reactors (PBRs) employing immobilized microorganisms. A low-cost, reusable immobilization matrix in PBR can provide economic advantages in large scale removal of high concentration phenol. In this study, we evaluated the efficiency and reusability of sugarcane bagasse (SCB) as a low-cost immobilization support for high strength phenol removal in recirculating upflow PBR. An isolated yeast Candida tropicalis PHB5 was immobilized onto the SCB support and packed into the reactor to assess phenol biodegradation at various influent flow rates. Scanning electron microscopy exhibited substantial cell attachment within the pith and onto the fibrous strand surface of the SCB support. The PBR showed 97% removal efficiency at the initial phenol concentration of 2400 mg L-1 and 4 mL min-1 flow rate within 54 h. Biodegradation kinetic studies revealed that the phenol biodegradation rate and biodegradation rate constant were dependent on the influent flow rate. A relatively higher rate of biodegradation (64.20 mg g-1 h-1) was found at a flow rate of 8 mL min-1, indicating rapid phenol removal in the PBR. Up to six successive batches (phenol removal >94%) were successfully applied in the PBR using an initial phenol concentration of 400-2400 mg L-1 at a flow rate of 4 mL min-1 indicating the reusability of the PBR system. The SCB-immobilized C. tropicalis could be employed as a cost-effective packing material for removal of high strength phenolic compounds in real scale PBR.

Improved dewatering of CEPT sludge by biogenic flocculant from Acidithiobacillus ferrooxidans.

Bioleaching using an iron-oxidizing bacterium, Acidithiobacillus ferrooxidans, and its biogenic flocculants was evaluated to improve the dewaterability of chemically enhanced primary treatment (CEPT) sewage sludge. CEPT sludge in flasks was inoculated with A. ferrooxidans culture, medium-free cells and the cell-free culture filtrate with and without the energy substance Fe(2+), and periodically the sludge samples were analysed for the dewaterability. This investigation proves that bioleaching effectively improved the sludge dewaterability as evidenced from drastic reduction in capillary suction time (≤20 seconds) and specific resistance to filtration (≥90%); however, it requires an adaptability period of 1-2 days. On the other hand, the biogenic flocculant produced by A. ferrooxidans greatly decreased the time-to-filtration and facilitated the dewaterability within 4 h. Results indicate that rapid dewatering of CEPT sludge by biogenic flocculants provides an opportunity to replace the synthetic organic polymer for dewatering.

Influence of clay minerals on sorption and bioreduction of arsenic under anoxic conditions.

Adsorption of As(V) on various clay minerals including kaolinite (KGa-1), montmorillonite (SWy-1) and nontronites (NAU-1 and NAU-2), and subsequent bioreduction of sorbed As(V) to As(III) by bacterium Shewanella putrefaciens strain CN-32 were investigated. Nontronites showed relatively higher sorption capacity for As(V) primarily due to higher iron oxide content. Freundlich equation well described the sorption of As(V) on NAU-1, NAU-2 and SWy-1, while As(V) sorption isotherm with KGa-1 fitted well in the Langmuir model. The bacterium rapidly reduced 50% of dissolved As(V) to As(III) in 2 h, followed by its complete reduction (>ca. 98%) within 12 h. In contrast, sorption of As(V) to the mineral surfaces interferes with the activity of bacterium, resulting in low bioreduction of As(V) by 27% for 5 days of incubation. S. putrefaciens also promoted the reduction of Fe(III) present in the clay mineral to Fe(II). This study indicates that the sorption and subsequent bioreduction of As(V) on clay minerals can significantly influence the mobility of As(V) in subsurface environment.

Degradation of a xenobiotic textile dye, Disperse Brown 118, by Brevibacillus laterosporus.

The toxic textile dye, Disperse Brown 118, was degraded by Brevibacillus laterosporus. 96% decolorization was achieved within 48 h at pH 7, 40 °C at 50 mg dye l(-1) accompanied by significant increases in the activities of veratryl alcohol oxidase, tyrosinase and NADH-DCIP reductase. HPTLC and FT-IR spectroscopy confirmed biodegradation after dye decolorization. As identified by GC-MS, biodegradation products of Disperse Brown 118 were N-carbamoyl-2-[(8-chloroquinazolin-4-yl)oxy] acetamide and N-carbamoyl-2-(quinazolin-4-yloxy)acetamide which were much less toxic than parent dye as evidenced by phytotoxicity tests.

Differential catalytic action of Brevibacillus laterosporus on two dissimilar azo dyes Remazol red and Rubine GFL.

This comparative study disclosed the diverse catalytic activities of Brevibacillus laterosporus on two different azo dyes. It decolorized 100% of Remazol red and 95% of Rubine GFL within 30 and 48 h respectively, under static condition at 50 mg l⁻¹ dye concentration. Significant increase was observed in azo reductase, NADH-DCIP reductase, veratryl alcohol oxidase and tyrosinase in cells obtained after decolorization of Remazol red; whereas these values were much different with complete inhibition of azo reductase during decolorization of Rubine GFL. The plausible pathway of dye degradation obtained from Gas chromatography-Mass spectroscopy (GC-MS) data confirmed the different metabolic fate of these structurally unidentical dyes. FTIR and HPTLC analysis of extracted metabolites confirmed the biodegradation, while phytotoxicity study assured the detoxification of both the dyes studied. The results obtained in this study suggests, i) sulpho and hydroxyl group present at ortho position to azo group stimulated reduction of azo bond by azo reductase in Remazol red, ii) the same reduction was totally hampered due to presence of ethyl-amino propanenitrile group at para position to azo group in Rubine GFL.

Fate of five bisphenol derivatives in Chlamydomonas mexicana: Toxicity, removal, biotransformation and microalgal metabolism.

Bisphenols (BPs) are recognized as emerging contaminants because of their estrogenic properties and frequent occurrence in environmental matrices. Here, we evaluated the toxic effects of five common BPs on freshwater microalga Chlamydomonas mexicana and removal of the BPs by the alga. Bisphenols -AF (BPAF), -B (BPB), and -Z (BPZ) (96 h, EC50 1.78-12.09 mg·L-1) exhibited higher toxicity to C. mexicana compared to bisphenol -S (BPS) and -F (BPF) (96 h, EC50 30.53-85.48 mg·L-1). In contrast, the mixture of BPs exhibited acute toxicity (96 h, EC50 8.07 mg·L-1). After 14 days, C. mexicana had effectively removed 61%, 99%, 55%, 87%, and 89% of BPS, BPF, BPAF, BPB, and BPZ, respectively, at 1 mg L-1. The biotransformed products of all five BPs were analyzed using UHPLC QTOF, and their toxicity was predicted. All biotransformed products were observed to be less toxic than the parent compounds. The fatty acid composition of C. mexicana after exposure to the BP mixture was predominantly palmitic acid (34.14%), followed by oleic acid (18.9%), and γ-linolenic acid (10.79%). The results provide crucial information on the ecotoxicity of these five BPs and their removal by C. mexicana; the resulting biomass is a potential feedstock for producing biodiesel.

Biochar as functional amendment for antibiotic resistant microbial community survival during hen manure composting.

The study aim was to reveal the mechanism of impact of two type biochar on composting of hen manure (HM) and wheat straw (WS). Biochar derived from coconut shell and bamboo used as additives to reduce antibiotic resistant bacteria (ARB) in HM compost. The results manifested that effect of biochar amendment was significant to reduce ARB in HM composting. Compared with control, the microbial activity and abundance were increased in both biochar applied treatment, and bacterial community was changed. Additionally, network analysis revealed that biochar amendment increased the quantity of microorganisms related to organic matter degrading. Among them, coconut shell biochar (CSB) played a pioneering role to mitigate ARB to better exert its effects. Structural correlation analysis showed that CSB reduce ARB mobility and promote organic matter degradation via improving beneficial bacterial community structure. Overall, composting with participation of biochar amendment stimulated antibiotic resistance bacterial dynamics. These results evidence practical value for scientific research and lay the foundation for agricultural promotion of composting.

Biodegradation of Rubine GFL by Galactomyces geotrichum MTCC 1360 and subsequent toxicological analysis by using cytotoxicity, genotoxicity and oxidative stress studies.

Galactomyces geotrichum MTCC 1360 showed 87 % decolorization of the azo dye Rubine GFL (50 mg l(-1)) within 96 h at 30 °C and pH 7.0 under static conditions, with significant reduction of chemical oxygen demand (67 %) and total organic carbon (59 %). Examination of oxidoreductive enzymes, namely laccase, tyrosinase and azo reductase, confirmed their role in decolorization and degradation of Rubine GFL. Biodegradation of Rubine GFL into different metabolites was confirmed using high-performance TLC, HPLC, Fourier transform IR spectroscopy and GC-MS analysis. During toxicological studies, cell death was observed in Rubine GFL-treated Allium cepa root cells. Toxicological studies before and after microbial treatment were done with respect to cytotoxicity, genotoxicity, oxidative stress, antioxidant enzyme status, protein oxidation and lipid peroxidation using root cells of A. cepa. The analysis with A. cepa showed that the dye exerts oxidative stress and subsequently has a toxic effect on the root cells, whereas its metabolites are less toxic. Phytotoxicity studies revealed the less toxic nature of the metabolites as compared with Rubine GFL.

Differential fate of metabolism of a disperse dye by microorganisms Galactomyces geotrichum and Brevibacillus laterosporus and their consortium GG-BL.

The present work aims to evaluate Brown 3 REL degrading potential of developed microbial consortium GG-BL using two microbial cultures, Galactomyces geotrichum MTCC 1360 (GG) and Brevibacillus laterosporus MTCC 2298 (BL). Microbial consortium GG-BL showed 100% decolorization of a dye Brown 3 REL, while individually G. geotrichum MTCC 1360 and B. laterosporus MTCC 2298 showed 26% and 86% decolorization under aerobic condition (shaking) respectively. Measurements of biochemical oxygen demand (BOD) (76%) and chemical oxygen demand (COD) (68%) were done after decolorization by consortium GG-BL. No induction in activities of oxidoreductive enzymes found in G. geotrichum while B. laterosporus showed induction of veratryl alcohol oxidase, Nicotineamide adenine dinucleotide-dichlorophenol indophenol (NADH-DCIP) reductase and riboflavin reductase indicating their role in dye metabolism. Consortium GG-BL showed induction in the activities of laccase, veratryl alcohol oxidase, tyrosinase, NADH-DCIP reductase and riboflavin reductase. Two different sets of induced enzymes from G. geotrichum and B. laterosporus work together in consortium GG-BL resulting in faster degradation of dye. The degradation of Brown 3 REL was analyzed using high performance thin layer chromatography (HPTLC), high performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR) and gas chromatography mass spectroscopy (GC-MS). Phytotoxicity study revealed that metabolites formed after degradation was significantly less toxic in nature.

A review on physico-chemical delignification as a pretreatment of lignocellulosic biomass for enhanced bioconversion.

Effective pretreatment of lignocellulosic biomass (LCB) is one of the most important steps in biorefinery, ensuring the quality and commercial viability of the overall bioprocess. Lignin recalcitrance in LCB is a major bottleneck in biological conversion as the polymerization of lignin with hemicellulose hinders enzyme accessibility and further bioconversion to fuels and chemicals. Therefore, there is a need to delignify LCB to ease further bioprocessing. The efficiency of delignification, quality and quantity of the desired products, and generation of inhibitors depend upon the type of pretreatment employed. This review summarizes different single and integrated physicochemical pretreatments for delignification. Additionally, conditions required for effective delignification and the advantages and drawbacks of each method were evaluated. Advances in overcoming the recalcitrance of residual lignin to saccharification and the methods to recover lignin after delignification are also discussed. Efficient lignin recovery and valorization strategies provide an avenue for the sustainable lignocellulose biorefinery.

Unravelling metabolism and microbial community of a phytobed co-planted with Typha angustifolia and Ipomoea aquatica for biodegradation of doxylamine from wastewater.

Pharmaceutical contaminants in environment induce unexpected effects on ecological systems and human; thus, development of efficient technologies for their removal is immensely necessary. In this study, biodegradation and metabolic fate of a frequently found pharmaceutical contaminant, doxylamine by Typha angustifolia and Ipomoea aquatica was investigated. Microbial community of the plant rhizosphere has been identified to understand the important roles of the functional microbes. The plants reduced 48-80.5 % of doxylamine through hydrolysis/dehydroxylation and carbonylation/decarbonylation. A constructed phytobed co-planted with T. angustifolia and I. aquatica removed 77.3 %, 100 %, 83.67 %, and 61.13 % of chemical oxygen demand, total nitrogen, total phosphorus, and doxylamine respectively from real wastewater. High-throughput sequencing of soil and rhizosphere indicated that the phyla Proteobacteria, Bacteroidetes, Firmicutes, Planctomycetes, Actinobacteria, and Cyanobacteria dominated the microbial communities of the phytobed. Current study has demonstrated the applicability of the developed phytobeds for the treatment of doxylamine from municipal wastewater and provide a comprehensive understanding of its metabolism through plant and its rhizospheric microbial communities.

Anaerobic co-digester microbiome during food waste valorization reveals Methanosaeta mediated methanogenesis with improved carbohydrate and lipid metabolism.

This study determines the optimum food waste (FW) loading in an anaerobic digester for methane production. Interrelation between the degradation mechanism and microbial community composition was assessed through in-depth metabolic pathway analysis and gene quantification. Higher methane production and short lag phase were observed in the FW reactors with low substrate loadings (<4% v/v) while extended lag phase and incomplete substrate utilization were observed in the reactors fed with higher substrates (>6% v/v). The long-chain fatty acids (LCFAs) degradation was influenced by initial FW loading, and up to 99% LCFA degradation occurred at 4% FW reactor. The addition of 8 to 10% FW substrate inhibited methanogenesis due to the accumulation of volatile fatty acids (VFA) and low LCFA degradation. Under optimal conditions of substrate loading, Methanosaeta and Methanosarcina were abundant, indicating their role in methanogenesis and syntrophic acetogenesis, along with enhanced metabolic pathways specific for carbohydrate and lipid metabolism.

Syntrophic metabolism facilitates Methanosarcina-led methanation in the anaerobic digestion of lipidic slaughterhouse waste.

Different inoculum to slaughterhouse waste (SHW) ratios (Ino/SHW) influences the digester performance, substrate utilization, and methane yield through microbial shift and their metabolic syntrophy. Acetoclastic Methanosarcina (68-87%) was dominant in the exponential phase, overpowering the initial abundance of Methanosaeta (86% of methanogens) in the SHW digesters. Positive interactions among acetogenic and acetate-oxidizing species of Clostridium (11%) with Methanosarcina (84% of methanogens) improved the methanogenic activity (292 mL g-1 VSinitial d-1) and final VS utilization (90%) at the highest Ino/SHW loading. In contrast, significant improvement of methane yield (152% higher than the control) at the lowest Ino/SHW loading was attributed to strong syntrophy among Methanosaeta (24% of methanogens) and its exoelectrogenic partners, Bythopirellula (0.52%) and Mariniphaga (0.08%) and the acetogenic Cloacimonas (0.16%) and Longilinea (0.32%). These syntrophic interactions among the core microbiota induced major metabolic activities, including butanoate, glycine, serine and threonine, methane, propanoate, and pyruvate metabolism, and quorum sensing.

Multiple metabolic pathways of enrofloxacin by Lolium perenne L.: Ecotoxicity, biodegradation, and key driven genes.

Contamination of fluoroquinolones (FQs) are of emerging concerns because of their adverse effects on environment and humans. This study investigated the ecotoxicological effects, biodegradation, and multiple metabolic pathways of a frequently found FQ, enrofloxacin (ENR) by ryegrass (Lolium perenne L.). Key metabolic genes for driving the metabolism of ENR have been identified using transcriptome profiling of L. perenne and gene network analysis. Toxicity of ENR on ryegrass has been evaluated according to the morphological changes, lipid peroxidation content, and antioxidant enzymatic activities. Moreover, there was 94.33%, 71.58%, 57.22%, and 55.23% removal of 1, 10, 50 and 100 mg L-1 ENR, respectively, which was mainly achieved by biodegradation according to the mass balance. A biodegradation pathway has been proposed by incorporating mass spectrums of extracted ENR intermediates with their formation dynamics. Analysis of differentially expressed genes (DEGs) and their network unraveled that the genes encoding monooxygenase, oxidative carboxylase, methyltransferase, lyase, hydroxylase, dehydrogenase, and peroxidase were the key functional genes. These enzymes can induce di/hydroxylation, decarboxylation, methylation, and bond and ring cleavage of ENR for its effective degradation. This study demonstrated that ryegrass can be used for efficient treatment of ENR polluted water and extended the understanding of the molecular mechanism of antibiotics' biodegradation in plants.

Rapid recovery of methane yield in organic overloaded-failed anaerobic digesters through bioaugmentation with acclimatized microbial consortium.

Acidification during anaerobic digestion (AD) due to organic overloading is one of the major reasons for process failures and decreased methane productivity in anaerobic digesters. Process failures can cause the anaerobic digesters to stall completely, prolong the digester recovery period, and inflict an increased operational cost on wastewater treatment plants and adverse impacts on the environment. This study investigated the efficacy of bioaugmentation by using acclimatized microbial consortium (AC) in recovering anaerobic digesters stalled due to acidosis. Overloading of digesters with food waste leachate (FWL) led to the accumulation of volatile fatty acids (11.30 g L-1) and a drop in pH (4.67), which resulted in process failure and a 22-fold decline in cumulative methane production compared to that in the initial phase. In the failure phase, the syntrophic and methanogenic activities of the anaerobic digester microbiota were disrupted by a significant decrease in the abundance of syntrophic populations such as Syntrophomonas, Syntrophorhabdus, Sedimentibacter, and Levilinea, and the phylum Euryarchaeota. Bioaugmentation of the failed digesters by adding AC along with the adjustment of pH resulted in the prompt recovery of methane productivity with a 15.7-fold higher yield than that in unaugmented control. The abundance of syntrophic bacteria Syntrophomonas and phylum Euryarchaeota significantly increased by 29- and 17-fold in the recovered digesters, respectively, which showed significant positive correlations with methane productivity. Methanosarcina and acetoclastic Methanosaeta played a major role in the recovery of the digesters; they were later replaced by hydrogenotrophic Methanoculleus. The increase in the abundance of genes associated with biomethanation contributed to digester recovery, according to the functional annotation of 16S rDNA amplicon data. Thus, bioaugmentation with AC could be a viable solution to recover digesters experiencing process failure due to organic overloading.

High-throughput integrated pretreatment strategies to convert high-solid loading microalgae into high-concentration biofuels.

The commercial feasibility of energy-efficient conversion of highly concentrated microalgal suspensions to produce high-titer biofuels is a major bottleneck due to high energy consumption. Herein, high-titer biofuels (bioethanol, higher-alcohols, and biodiesel) were generated from carbohydrate-rich Chlamydomonas mexicana and lipid-rich Chlamydomonas pitschmannii biomass through energy-saving microwave pretreatment, successive fermentation, and transesterification. Microwave pretreatment needed low specific energy (4.2 MJ/kg) for 100 g/L of microalgal suspension. Proposed sustainable integrated pretreatments method achieved unprecedented total conversion efficiency (67%) and highest biomass utilization (87%) of C. pitschmannii (100 g/L) with high yields of bioethanol (0.48 g-ethanol/g-carbohydrates), higher-alcohols (0.44 g-higher-alcohols/g-proteins), and biodiesel (0.90 g-biodiesel/g-lipids). Transmission electron microscopy showed the changes in the microalgal cellular integrity before and after sequential fermentations. Energy-efficient integrated pretreatments enhanced the extraction efficiency and whole utilization of high-concentration microalgae to generate high-titer biofuels with minimum waste production.

Microbial community acclimatization for enhancement in the methane productivity of anaerobic co-digestion of fats, oil, and grease.

The methane productivity and long chain fatty acids (LCFAs) degradation capability of unacclimatized seed sludge (USS) and acclimatized seed sludge (ASS) at different substrate ratios of fats oil and grease (FOG) and mixed sewage sludge were investigated in this study. Biogas produced in ASS in initial phase of anaerobic digestion had higher methane content (65-76%) than that in USS (26-73%). The degradation of major LCFAs in the ASS was 22-80%, 33-191%, and 7-64% higher for the substrate ratios of 100:10, 100:20, and 100:30, respectively, as compared to the LCFAs' degradation in USS. Microbial acclimatization increased the population of Firmicutes (40%), Bacteroidetes (32%), Synergistetes (10%), and Euryarchaeota (8%) in ASS, which supported the faster rate of LCFAs degradation for its later conversion to methane. The significant abundance of Syntrophomonas and Methanosarcina genera in ASS supported faster generation rate of methane in an obligatory syntrophic relationship.