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The commercial production of bioethanol from lignocellulosic biomass is challenged by the repression of cell growth and compromised fermentation conditions. However, employing Saccharomyces cerevisiae is an economically feasible process resulting in the effective conversion of fermentable sugars to bioethanol. S. cerevisiae’s high productive feature is contributed by robust alcohol dehydrogenase and the ability to tolerate various stresses during the fermentation process. The present review employs various bioinformatics pipeline to assess the structural insights of ADH and interaction among the tolerance genes. Docking of S. cerevisiae’s ADH interaction shows the high binding affinity of − 2.81 Kcal/mol with acetaldehyde contributed by four zinc fingers. In inhibitor tolerance capacity of S. cerevisiae was explored. The STRING platform sheds light on the mechanism and interaction of ASR1 and other stress-responsive elements playing a pivotal role in ethanol tolerance. The stress-responsive genes such as HSPs and MSN2/4 provide balanced physiology under various stress conditions. The present review unravels the complex mechanism behind the inhibitor and ethanol tolerance, directing to several bottlenecks for the improvisation of S. cerevisiae’s performance.

Recently discovered Lytic Polysaccharide Mono-Oxygenase (LPMO) enhances the enzymatic deconstruction of complex polysaccharide by oxidation. The present study demonstrates the agricultural waste hydrolyzing capabilities of Paenibacillus dendritiformis CRN18, which exhibits the enzyme activity of exo-glucanase, β-glucosidase, β-glucuronidase, endo-1, 4 β-xylanases, arabinosidase, and α–galactosidase as 0.1U/ml, 0.3U/ml, 0.09U/ml, 0.1U/ml, 0.05U/ml, and 0.41U/ml, respectively. The genome analysis of strain reveals the presence of four LPMO genes, along with lignocellulolytic genes. The gene structure of LPMO and its phylogenetic analysis shows the evolutionary relatedness with the Bacillus LPMO gene. Gene position of LPMOs in the genome of strains shows the close association of two LPMOs with chitin active enzyme GH18, and the other two are associated with hemicellulases (GH39, GH23). Protein-protein interaction and gene networking of LPMO sheds light on the co-occurrence, neighborhood, and interaction of LPMOs with chitinase and xylanase enzymes. Structural prediction of LPMOs unravels the information of the LPMO's binding site. Although the LPMO has been explored for its oxidative mechanism, a little light has been shed on its gene structure. This study provides insights into the LPMO gene structure in P. dendritiformis CRN18 and its potential in lignocellulose hydrolysis.

Prophylactic usage and high persistent nature of several antibiotics have put selective pressure on the native microbial population that led to the emergence, propagation, and persistence of antibiotic resistance in nature. The surveillance of antibiotic resistome pattern and identification of points of intervention throughout the different environmental habitats will help to break the flow of antibiotic resistance from environmental bacteria to human pathogens. The present study compares the occurrence, diversity, and abundance of ARGs in industrial sludge, wetland sludge, and sediment sample contaminated with pharmaceutical discharge. Metagenomes were mined for the presence of ARGs against the ResFinder 3.2 database using BLASTn program. Pharmaceutical sample (2.52%) showed high degree of ARG abundance and richness as compared with ETP sludge (2.28%) and wetland sludge samples (1.29%). The modern resistome pattern represented by critically important resistance genes against tetracycline (tetA, tetC, tetW, tetT, and tetS/M) and quinolone (qnrS, qnrVC, and qnrD) was identified in pharmaceutical sediment sample. However, effluent treatment plant (ETP) sludge sample showed abundance of multidrug efflux pumps indicating the presence of primitive resistome profile. In conclusion, the indiscriminate distribution pattern of antibiotic resistance genes in three selected environmental sites suggests enrichment and distribution of environmental niche–driven resistance. The study also suggests effluent discharge site from pharmaceutical industries and ETPs as pivotal points of intervention for the mitigation of antibiotic resistance.

Cellulose is the most abundant natural polymer present on Earth in the form of agriculture waste. Hydrolysis of agriculture waste for simple fermentable reducing sugars is the bottleneck in the area of biofuel generation and other value-added products. The present study aims to utilize the camel rumen as a bioreactor for potent cellulolytic and hemicellulolytic bacteria by altering the feed types with varying cellulosic concentrations. A total of 6716 bacterial cultures were subjected to three layers of screening, where plate zymography and chromophoric substrate screening served as primary screening method for cellulolytic and hemicellulolytic potential. The potential isolates were genetically grouped using RAPD, and 51 representative isolates from each group were subjected to molecular identification through 16S rDNA sequencing, followed by quantification of various cellulolytic and hemicellulolytic enzymes. Out of 51 potent isolates, 5 isolates had high endoglucanase activity ranging from 0.3 to 0.48 U/ml. The selected five key isolates identified as Pseudomonas, Paenibacillus, Citrobacter, Bacillus subtilis, and Enterobacter were employed for hydrolyzing the various agriculture residues and resulted in approximately 0.4 mg/ml of reducing sugar. Furthermore, the metaculturomics approach was implemented to deduce the total cultured diversity through 16S rRNA amplicon library sequencing. The metaculturomics data revealed the dominance of proteobacteria and unidentified bacterial population in all four feed types, which indicates the possibility of culturing novel cellulose-deconstructing bacteria. Moreover, the presence of diverse hydrolytic enzymes in cultured isolates supports the usage of these bacteria in bio-processing of agriculture waste residues and obtaining the biofuels and other value-added products.

Inefficient sewage management has caused contamination of our water resources (rivers and lakes) that inspires researchers to constantly innovate and upgrade the existing wastewater treatment technologies. This study explores the possibility of a microbial consortium immobilized on different natural matrix for sewage treatment at lab-scale bioreactor. Bacterial isolates having lipolytic, proteolytic, cellulolytic and amylolytic activities were included in consortia as these are the major constitutes of sewage. The activities of these bacterial cultures were confirmed using different microbiological and molecular tools. The microbial consortium immobilized on natural coconut coir was found to be the most effective matrix to enhance the efficiency of sewage treatment. The immobilization of microbes on a natural dry coconut coir matrix was confirmed through scanning electron microscopy (SEM). Two Lab-scale aerobic bioreactors were setup in parallel for the treatment of sewage and simulated wastewaters. Both the reactors were operated in continuous mode for 15 and 16 days and efficiency of treatment was analysed by monitoring chemical oxygen demand (COD) removal, nitrogen removal as NH4-N and NO3-N. Simultaneously, the nitrogen removal activity was validated in all members of microbial consortium using gene-specific primers. Results show that 75.9% and 73.7% COD removal were observed in column C and D and nitrogen as NO3-N 78.9% and 79.7% (NH4-N) removal were measured in the effluent of reactors. Similarly, 93% COD, 78.1% NO3-N and 79.7% NH4-N removal were observed in a stimulated wastewater reactor. This significant removal of organic material shows the higher efficiency of the reactor which could be used for field application for sewage treatment.

In addition to a wide variety of anaerobic and facultative anaerobic bacteria, camel rumen also harbors a diverse of eukaryotic organisms. In the present study, the eukaryotic communities of camel rumen were characterized using 18S rRNA amplicon sequencing. Metagenomic DNA was isolated from rumen samples of fourteen adult Bikaneri and Kachchhi breeds of camel fed different diets containing Jowar, Bajra, Maize, and Guar. Illumina sequencing generated 27,161,904 number of reads corresponding to 1543 total operational taxonomic units (OTUs). Taxonomic classification of community metagenome sequences from all the samples revealed the presence of 92 genera belonging to 16 different divisions, out of which Ciliophora (73%), Fungi (13%) and Streptophyta (9%) were found to be the most dominant. Notably, the abundance of Ciliophora was significantly higher in the case of Guar feed, while Fungi was significantly higher in the case of Maize feed, indicating the influence of cellulose and hemicellulose content of feedstuff on the composition of eukaryotes. The results suggest that the camel rumen eukaryotes are highly dynamic and depend on the type of diet given to the animal. Pearson's correlation analysis suggested the ciliate protozoa and fungi were negatively correlated with each other. To the best of our knowledge, this is first systematic study to characterize camel rumen eukaryotes, which has provided newer information regarding eukaryotic diversity patterns amongst camel fed on different diets.

Microbes play a major role in phosphate cycling and regulate its availability in various environments. The metagenomic study highlights the microbial community divergence and interplay of phosphate metabolism functional genes in response to phosphate rich (100 mgL

Reconstruction of genomes from environmental metagenomes offers an excellent prospect for studying the metabolic potential of organisms resilient to isolation in laboratory conditions. Here, we assembled 12 high-quality metagenome-assembled genomes (MAGs) with an estimated completion of ≥ 90% from cow and buffalo rumen metagenomes. Average nucleotide identity (ANI) score-based screening with an existing database suggests the novelty of these genomes. Gene prediction led to the identification of 30,359 protein-encoding genes (PEGs) across 12 genomes, of which only 44.8% were annotated against a specific functional attribute. Further analysis revealed the presence of 985 carbohydrate-active enzymes (CAZymes) from more than 50 glycoside hydrolase families, of which 90% do not have a proper match in the CAZy database. Genome mining revealed the presence of a high frequency of plant biomass deconstructing genes in Bacteroidetes MAGs compared to Firmicutes. The results strongly indicate that the rumen chamber harbors high numbers of deeply branched and as-yet uncultured microbes that encode novel CAZymes, candidates for prospective usage in plant biomass-hydrolyzing and biofuels industries. KEY POINTS: • Genome binning plays a crucial role in revealing the metabolic potential of uncultivable microbes. • Assembled 12 novel genomes from cow and buffalo rumen metagenome datasets. • High frequency of plant biomass deconstructing genes identified in Bacteroidetes MAGs.

The major reason behind the spread of antibiotic resistance genes (ARGs) is persistent selective pressure in the environment encountered by bacteria. Genome plasticity plays a crucial role in dissemination of antibiotic resistance among bacterial pathogens. Mobile genetic elements harboring ARGs are reported to dodge bacterial immune system and mediate horizontal gene transfer (HGT) under selective pressure. Residual antibiotic pollutants develop selective pressures that force the bacteria to lose their defense mechanisms (CRISPR-cas) and acquire resistance. The present study targets the ESKAPE organisms (namely, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) causing various nosocomial infections and emerging multidrug-resistant species. The role of CRISPR-cas systems in inhibition of HGT in prokaryotes and its loss due to presence of various stressors in the environment is also focused in the study. IncF and IncH plasmids were identified in all strains of E. faecalis and K. pneumoniae, carrying Beta-lactam and fluoroquinolone resistance genes, whereas sal3, phiCTX, and SEN34 prophages harbored aminoglycoside resistance genes (aadA, aac). Various MGEs present in selected environmental niches that aid the bacterial genome plasticity and transfer of ARGs contributing to its spread are also identified.

Genome sequencing of Klebsiella pneumoniae PVN-1, isolated from effluent treatment plant (ETP), generates a 5.064 Mb draft genome with 57.6% GC content. The draft genome assembled into 19 contigs comprises 4783 proteins, 3 rRNA, 44 tRNA, 8 other RNA, 4911 genes, and 73 pseudogenes. Genome information revealed the presence of phosphate metabolism/solubilizing, potassium solubilizing, auxin production, and other plant benefiting attributes like enterobactin and pyrroloquinoline quinone biosynthesis genes. Presence of gcd and pqq genes in K. pneumoniae PVN-1 genome validates the inorganic phosphate solubilizing potential (528.5 mg/L). Pangenome analysis identified a unique 5ʹ-Nucleotidase that further assists in enhanced phosphate acquisition. Additionally, the genetic potential for complete benzoate, catechol, and phenylacetate degradation with stress response and heavy metal (Cu, Zn, Ni, Co) resistance was identified in K. pneumoniae PVN-1. Functioning of annotated plant benefiting genes validates by the metabolic activity of auxin production (7.40 µg/mL), nitrogen fixation, catalase activity, potassium solubilization (solubilization index—3.47), and protease activity (proteolytic index—2.27). In conclusion, the K. pneumoniae PVN-1 genome has numerous beneficial qualities that can be employed to enhance plant growth as well as for phytoremediation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03020-2.

Rumen microbial community harbors a distinct genetic reservoir of potent carbohydrate-active enzymes (CAZyme) that functions efficiently for the deconstruction of plant biomass. Based on this premise, metagenomics approach was applied to characterize the rumen microbial community and identify carbohydrate-active genes of Bos taurus (cow) and Bubalus bubalis (buffalo) fed on green or dry roughage. Metadata was generated from the samples: green roughage-fed cow (NDC_GR), buffalo (NDB_GR) and dry roughage-fed cow (NDC_DR), buffalo (NDB_DR). Phylogenetic analysis revealed the dominance of Bacteroidetes, Firmicutes, Proteobacteria, Actinobacteria and Fibrobacter in all the four samples, covering 90-96% of the total bacterial population. On finer resolution, higher abundance of bacterial genera Fibrobacter, Bacteroides, Clostridium, Prevotella and Ruminococcus involved in plant biomass hydrolysis was observed in NDB_DR. Functional annotation using dbCAN annotation algorithm identified 28.13%, 8.08% 10.93% and 12.53% of the total contigs as putatively carbohydrate-active against NDC_GR, NDB_GR, NDC_DR and NDB_DR, respectively. Additional profiling of CAZymes revealed an over representation and diversity of putative glycoside hydrolases (GHs) in the animals fed on dry roughage with substantial enrichments of genes encoding GHs from families GH2, GH3, GH13 and GH43. GHs of families GH45, GH12, GH113, GH128, GH54 and GH27 were observed exclusively in NDB_DR metagenome. A higher abundance of cellulases, hemicellulases, debranching and oligosaccharide hydrolyzing enzymes was revealed in NDB_DR metagenome. Accordingly, it can be concluded that buffalo rumen microbiome are more efficient in plant biomass hydrolysis. The present study provides a deep understanding of the shifts in microbial community and plant polysaccharide deconstructing capabilities of rumen microbiome in response to changes in the feed type and host animal. Activity-specific microbial consortia procured from these animals can be used further for efficient plant biomass hydrolysis. The study also establishes the utility of rumen microbiome as a unique resource for mining diverse lignocellulolytic enzymes.

This study aims at designing a consortium using rumen bacterial isolates for enhancing the hydrolysis of sugarcane bagasse (SB) for efficient biofuel formation. The microbial population was screened through biochemical and molecular tools along with enzymatic activity to obtain potential isolates for diverse cellulolytic and hemicellulolytic carbohydrate active enzyme (CAZyme). Five strains (Paenibacillus, Bacillus, Enterobacter, and Microbacterium) were selected for designing the consortium NDMC-1. The hydrolytic efficiency of NDMC-1 was determined based on cellulase production with simultaneous rise in monosaccharides, oligosaccharides, and soluble chemical oxygen demand (sCOD) concentration. Cellulolytic machinery of these isolates was further explored using genome sequencing. The isolates selected for consortia NDMC-1 interacted synergistically leading to enhanced cellulase production. Maximal endoglucanase (1.67 μmol ml−1 min−1), exoglucanase (0.69 μmol ml−1 min−1), and β-glucosidase (2.03 μmol ml−1 min−1) activity were achieved with SB as a sole carbon source after 48 h of incubation. Enhancement in SB hydrolysis employing NDMC-1 was evident by the increase in sCOD from 609 to 2589 mg/l and release of 1295 mg/l reducing sugar, comprising 59.8%, 8.23%, and 6.16% of glucose, cellobiose, and cellotriose, respectively, which resulted in 5.5-fold rise in biogas production. On genome annotation, total 472 contigs from glycoside hydrolase family: 84 from Microbacterium arborescens ND21, 72 from Enterobacter cloacae ND22, 61 from Bacillus subtilis ND23, 116 from Paenibacillus polymyxa ND24, and 140 from Paenibacillus polymyxa ND25 were identified. On further analysis, total 33 cellulases, 59 hemicellulases, and 48 esterases were annotated in the reported genomes. This work proposes the application of consortia-based bioprocessing systems over the conventionally favorable single organism approach for efficient hydrolysis of cellulosic substrates to fermentable sugars.

Improving enzymatic saccharification of lignocellulosic biomass is the major bottle neck in the area of second generation biofuel production. Metalloenzyme lytic polysaccharide monooxygenase (LPMO) has the capability to oxidize the complex lignocellulose in the crystalline area and thus acts as a catalyst in the process of lignocellulosic deconstruction. The present review undermines the importance of LPMO in biofuel sectors. It emphasizes evolutionary relationship, structure, mechanism, and synergistic action of LPMOs. A radial phylogram of 168 LPMO genes from archaea to terrestrial plant infers their evolution in different domains. The existence of LPMOs in extreme environmental condition and structural insights of these LPMOs show the conservation in active copper center and flat surface for substrate binding. This finding may open a new way for the isolation of active LPMOs from different environmental condition and their optimization to withstand the industrial parameters. The synergistic action of LPMO with different cellulase and hemicellulase may help in lowering the catalytic load which ultimately may reduce the cost of pretreatment steps. This review therefore sums up the recent findings concerning the LPMO’s mechanism of action and identifies issues and questions to be addressed in the future.

In India, sewage (partially-treated/ untreated) is randomly used for irrigation because of easy availability and presence of residual organics and nutrients. However, data on the occurrence of contaminants of emerging concerns (CECs) such as pharmaceuticals and personal care products (PPCPs) and antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs) in sewage is scarce in Indian perspective. Herein, for the first time, we present a quantitative contamination profiling of selected PPCPs and antibiotic resistance in untreated and biologically-treated sewage from three different sewage treatment plants, located in northern and central part of India. Profiling of PPCPs were done using LC-ESI-MS/MS whereas antibiotic resistance was analyzed using gradient PCR and qPCR techniques. PPCPs were detected both in untreated and treated samples (0.4 – 1340 μg/L). A reduction in ARB and ARG load (2–3 log) and an increase in ARG copy number with respect to beta lactams and tetracycline were observed in treated sewage. Triclosan, estrone and 17α–ethynylestradiol, ubiquitous in all samples, could be used as markers for performance monitoring of sewage treatment facilities. The results obtained in this study help evaluate health and ecological risks associated with the presence of CECs in treated sewage used for irrigation and frame future policies.

Prophylactic usage and high persistent nature of several antibiotics have put selective pressure on the native microbial population that led to the emergence, propagation, and persistence of antibiotic resistance in nature. The surveillance of antibiotic resistome pattern and identification of points of intervention throughout the different environmental habitats will help to break the flow of antibiotic resistance from environmental bacteria to human pathogens. The present study compares the occurrence, diversity, and abundance of ARGs in industrial sludge, wetland sludge, and sediment sample contaminated with pharmaceutical discharge. Metagenomes were mined for the presence of ARGs against the ResFinder 3.2 database using BLASTn program. Pharmaceutical sample (2.52%) showed high degree of ARG abundance and richness as compared with ETP sludge (2.28%) and wetland sludge samples (1.29%). The modern resistome pattern represented by critically important resistance genes against tetracycline (tetA, tetC, tetW, tetT, and tetS/M) and quinolone (qnrS, qnrVC, and qnrD) was identified in pharmaceutical sediment sample. However, effluent treatment plant (ETP) sludge sample showed abundance of multidrug efflux pumps indicating the presence of primitive resistome profile. In conclusion, the indiscriminate distribution pattern of antibiotic resistance genes in three selected environmental sites suggests enrichment and distribution of environmental niche-driven resistance. The study also suggests effluent discharge site from pharmaceutical industries and ETPs as pivotal points of intervention for the mitigation of antibiotic resistance.

Bacterial esterases are gaining the importance in pharmaceuticals and agrochemical industries due to their excellent biocatalytic properties and a wide range of applications. In the present study, a novel gene encoding an esterase (designated as Est-CR) was identified from shotgun metagenomic sequencing data of camel rumen (Camelus dromedarius) liquor. The open reading frame consisted of 1,224bp, which showed 84.03% sequence identity to Bacteroidales bacterium, corresponding to a protein of 407 amino acids and has a catalytic domain belonging to an esterase. Est-CR belonged to family V with GLSMG domain. The purified enzyme with a molecular mass of 62.64 kDa was checked on SDS-PAGE, and its expression was confirmed by western blotting. The enzyme was active and stable over a broad range of temperature (35–65 °C), displayed the maximum activity at 50 °C and pH 7.0. Individually all metal ions inhibited the enzyme activity, while in combination, K2+, Ca2+, Mg2+ and Mn2+ metal ions enhanced the enzyme activity. The detergents strongly inhibited the activity, while EDTA (10 mM) increased the activity of the Est-CR enzyme. The enzyme showed specificity to short-chain substrates and displayed an optimum activity against butyrate ester. This novel enzyme might serve as a promising candidate to meet some harsh industrial processes enzymatic needs.

The bacterial communities are challenged with oxidative stress during their exposure to bactericidal antibiotics, metals, and different levels of dissolved oxygen (DO) encountered in diverse environmental habitats. The frequency of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) co-selection is increased by selective pressure posed by oxidative stress. Hence, study of resistance acquisition is important from an evolutionary perspective. To understand the dependence of oxidative stress on the dissemination of ARGs and MRGs through a pathogenic bacterial population, 12 metagenomes belonging to gut, water and soil habitats were evaluated. The metagenome–wide analysis showed the chicken gut to pose the most diverse pool of ARGs (30.4 ppm) and pathogenic bacteria (Simpson diversity = 0.98). The most common types of resistances found in all the environmental samples were efflux pumps (13.22 ppm) and genes conferring resistance to vancomycin (12.4 ppm), tetracycline (12.1 ppm), or beta-lactam (9.4 ppm) antibiotics. Additionally, limiting DO level in soil was observed to increase the abundance of excision nucleases (uvrA and uvrB), DNA polymerase (polA), catalases (katG), and other oxidative stress response genes (OSGs). This was further evident from major variations occurred in antibiotic efflux genes due to the effect of DO concentration on two human pathogens, namely Salmonella enterica and Shigella sonnei found in all the selected habitats. In conclusion, the microbial community, when challenged with oxidative stress caused by environmental variations in oxygen level, tends to accumulate higher amounts of ARGs with increased dissemination potential through triggering non-lethal mutagenesis. Furthermore, the genetic linkage or co-occurrence of ARGs and MRGs provides evidence for selecting ARGs under high concentrations of heavy metals.

To explore the diversity of bacterial endophytes of rhizomes of bamboo growing in the central part of India; five Bamboo species were collected and investigated. The culturable endophytic bacteria were isolated by serial dilution and culture-dependant method and their diversity was studied with a 16S rDNA sequence analysis. Consequently, a total of 31 endophytic bacteria isolated from these bamboo species, and the isolates were affiliated to 17 different genera. Proteobacteria was the dominant phylum with the highest diversity containing 11 genera and Staphylococcus, Herbaspirillum, Pseudomonas and Stenotrophomonas being the most frequently observed genera. The Microbacterium, Chryseobacterium, and Chitinophaga were the solitary representative of classes Actinobacteria, Flavobacteria, and Sphingobacteria, respectively. The diversity of bacteria was greater in Dendrocalamus species than in Bambusa species. Based on the putative identity of isolates and their functional roles as reported in the literature, the isolated bacterial endophytes seems to be the promising sustainable bioresource for bioaugmentation of plant growth particularly bamboos in tropical areas including forest.

An environment friendly and cost-effective approach is a major bottleneck in biofuels production from agriculture waste residue. The present study unravels the enzymatic hydrolytic potential of a designed bacterial consortium (SNH-1) comprised of camel rumen derived B. safensis CRN 13, B. subtilis CRN 16, C. braakii CRN 21, B. circulans CRN 24, and P. dendritiformis CRN18. The consortium SNH-1 was evaluated for exo-glucanase (0.29 U/ml), β-glucosidase (0.69U/ml), β-glucuronidase (0.29U/ml), endoxylanase (0.79U/ml), arabinosidase (0.62U/ml), α-galactosidase (0.36 U/ml), and endoglucanase (0.51U/ml) activity. Genome analysis of consortium members identified 660 carbohydrate-active enzymes (CAZyme) responsible for breaking the structural intricacy. These include 296 glycoside hydrolase, 19 auxiliary activity, 186 glycosyl transferases, and 37 carbohydrate-binding modules genes. The designed consortium hydrolyzed sugarcane bagasse, wheat straw, and rice straw effectively. Response surface methodology (RSM) was applied to optimize the pH, temperature, and substrate concentration, which has resulted in a 23% higher saccharification of wheat straw. Further, 6.2% bioethanol production was estimated from wheat straw hydrolysate using S. cerevisiae. The wheat straw saccharification by the designed consortium is a cost-effective, chemical-free, cleaner, and promising approach for producing lignocellulosic bioethanol.

The spread of multidrug-resistant (MDR) bacteria is an emerging threat to the environment and public wellness. Inappropriate use and indiscriminate release of antibiotics in the environment through un-metabolized form create a scenario for the emergence of virulent pathogens and MDR bugs in the surroundings. Mechanisms underlying the spread of resistance include horizontal and vertical gene transfers causing the transmittance of MDR genes packed in different host, which pass across different food webs. Several controlling agents have been used for combating pathogens; however, the use of lytic bacteriophages proves to be one of the most eco-friendly due to their specificity, killing only target bacteria without damaging the indigenous beneficial flora of the habitat. Phages are part of the natural microflora present in different environmental niches and are remarkably stable in the environment. Diverse range of phage products, such as phage enzymes, phage peptides having antimicrobial properties, and phage cocktails also have been used to eradicate pathogens along with whole phages. Recently, the ability of phages to control pathogens has extended from the different areas of medicine, agriculture, aquaculture, food industry, and into the environment. To avoid the arrival of pre-antibiotic epoch, phage intervention proves to be a potential option to eradicate harmful pathogens generated by the MDR gene flow which are uneasy to cure by conventional treatments.

Pharmaceuticals and personal care products are diverse compounds used for the treatment of diseases to promote health, enhance growth and meat production in farm animals. The consumption of pharmaceuticals has increased by 36% during the year 2000–10. Pharmaceuticals are often excreted in unmetabolized form or as active metabolites into the environment. These emerging contaminants impose selection pressure on the environmental microbes leading to the development of antibiotic resistance. Microbes have now gained resistance against “the last resort of antibiotics.” Horizontal gene transfer plays the major role in the dissemination of antimicrobial resistance in diverse niches. It accounts for about 70% exchange of resistance genes between pathogens and environmental microbes. A global, multidimensional approach is required to combat antibiotic resistance. It should emphasize on the conservation of antibiotics by judicious use, adoption of hygiene and cleaner production technology, vaccine development against diseases, and the discovery of novel antimicrobials compounds using functional metagenomics and CRISPR-CAS techniques.

Pho regulon is a highly evolved and conserved mechanism across the microbes to fulfil their phosphate need. In this study, 52 proteobacteria genomes were analyzed for the presence of phosphorus acquisition genes, their pattern of arrangement and copy numbers. The diverse genetic architecture of the Pho regulon genes indicates the evolutionary challenge of nutrient limitation, particularly phosphorus, faced by bacteria in their environment. The incongruence between the Pho regulon proteins phylogeny and species phylogeny along with the presence of additional copies of pstS and pstB genes, having cross similarity with other genera, suggest the possibility of horizontal gene transfer event. The substitution rate analysis and multiple sequence alignment of the Pho regulon proteins were analyzed to gain additional insight into the evolution of the Pho regulon system. This comprehensive study confirms that genes perform the regulatory function (phoBR) were vertically inherited, whereas interestingly, genes whose product involved in direct interaction with the environment (pstS) acquired by horizontal gene transfer. The substantial amino acid substitutions in PstS most likely contribute to the successful adaptation of bacteria in different ecological condition dealing with different phosphorus availability. The findings decipher the intelligence of the bacteria which enable them to carry out the targeted alteration of genes to cope up with the environmental condition.

Genome sequence of Paenibacillus polymyxa ND25 isolated from cow rumen is reported for being a potential candidate in hydrolysis of lignocellulosic plant biomass. Draft genome sequence generated 5.73 Mb data containing 4922 putative protein coding genes, of which 140 are annotated for glycoside hydrolases. P. polymyxa ND25 strain comprises diverse lignocellulolytic components, especially 12 cellulase along with 23 hemicellulases and 11 esterases, signifying its potential for lignocellulose hydrolysis. Subsequent enzyme assay exhibited the potential of strain to produce 0.49, 0.24 and 0.44 U/ml U/ml of endoglucanase, exoglucanase and β-glucosidase, respectively, utilizing sugarcane bagasse as the sole carbon source. This study signifies the efficient application of P. polymyxa ND25 for facilitating plant-biomass utilization. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-018-1274-3) contains supplementary material, which is available to authorized users.

Cellulolytic bacteria from cattle rumen with ability to hydrolyze cellulose rich biomass were explored. The study selected Paenibacillus polymyxa ND24 from 847 isolates as the most potent strain, which can efficiently produce cellulase by utilizing sugarcane bagasse, rice straw, corn starch, CMC, and avicel as a sole carbon source. On annotation of P. polymyxa ND24 genome, 116 members of glycoside hydrolase (GH) family from CAZy clusters were identified and the presence of 10 potential cellulases was validated using protein folding information. Cellulase production was further demonstrated at lab-scale 5-L bioreactor exhibiting maximum endoglucanase activity up to 0.72 U/mL when cultivated in the medium containing bagasse (2% w/v) after 72 h. The bagasse hydrolysate so produced was further utilized for efficient biogas production. The presence of diverse hydrolytic enzymes and formidable cellulase activity supports the use of P. polymyxa ND24 for cost-effective bioprocessing of cellulosic biomass.

Combating bacterial pathogens has become a global concern especially when the antibiotics and chemical agents are failing to control the spread due to its resistance. Bacteriophages act as a safe biocontrol agent by selectively lysing the bacterial pathogens without affecting the natural beneficial microflora. The present study describes the screening of prominent enteric pathogens NDK1, NDK2, NDK3, and NDK4 (Escherichia, Klebsiella, Enterobacter, and Serratia) mostly observed in domestic wastewater; against which KNP1, KNP2, KNP3, and KNP4 phages were isolated. To analyze their potential role in eradicating enteric pathogens and toxicity issue, these bacteriophages were sequenced using next-generation sequencing and characterized based on its genomic content. The isolated bacteriophages were homologous to Escherichia phage (KNP1), Klebsiella phage (KNP2), Enterobacter phage (KNP3), Serratia phage (KNP4), and belonged to Myoviridae family of Caudovirales except for the unclassified KNP4 phage. Draft genome analysis revealed the presence of lytic enzymes such as holing and lysozyme in KNP1 phage, endolysin in KNP2 phage, and endopeptidase with holin in KNP3 phage. The absence of any lysogenic and virulent genes makes this bacteriophage suitable candidate for preparation of phage cocktail to combat the pathogens present in wastewater. However, KNP4 contained a virulent gene rendering it unsuitable to be used as a biocontrol agent. These findings make the phages (KNP1–KNP3) as a promising alternative for the biocontrol of pathogens in wastewater which is the main culprit to spread these dominated pathogens in different natural water bodies. This study also necessitates for genomic screening of bacteriophages for lysogenic and virulence genes prior to its use as a biocontrol agent.

Bacteriophages play significant role in driving microbial diversity; however, little is known about the diversity of phages in different ecosystems. A dynamic predator-prey mechanism called "kill the winner" suggests the elimination of most active bacterial populations through phages. Thus, interaction between phage and host has an effect on the composition of microbial communities in ecosystems. In this study, secondary phage metagenome data from aquatic habitats: wastewater treatment plant (WWTP), fresh, marine, and hot water spring habitat were analyzed using MG-RAST and STAMP tools to explore the diversity of the viruses. Differential relative abundance of phage families-Siphoviridae (34%) and Myoviridae (26%) in WWTP, Myoviridae (30%) and Podoviridae (23%) in fresh water, and Myoviridae (41%) and Podoviridae (8%) in marine-was found to be a discriminating factor among four habitats while Rudiviridae (9%), Globuloviridae (8%), and Lipothrixviridae (1%) were exclusively observed in hot water spring. Subsequently, at genera level, Bpp-1-like virus, Chlorovirus, and T4-like virus were found abundant in WWTP, fresh, and marine habitat, respectively. PCA analysis revealed completely disparate composition of phage in hot water spring from other three ecosystems. Similar analysis of relative abundance of functional features corroborated observations from taxa analysis. Functional features corresponding to phage packaging machinery, replication, integration and excision, and gene transfer discriminated among four habitats. The comparative metagenomics approach exhibited genetically distinct phage communities among four habitats. Results revealed that selective distribution of phage communities would help in understanding the role of phages in food chains, nutrient cycling, and microbial ecology. Study of specific phages would also help in controlling environmental pathogens including MDR bacterial populations using phage therapy approach by selective mining and isolation of phages against specific pathogens persisting in a given environment.

The phosphorus availability in soil ranged from

Increasing levels of carbon dioxide (CO2) in the atmosphere are causing serious effect on climatic changes. Thereby the most concerned environmental issue is global warming these days. The same culprit carbon dioxide is also involved in the most important process called autotrophy, which supports life on earth. Autotrophy could be one of the probable answers for the management of atmospheric CO2 in an eco-friendly manner. Plants contribute a major part in CO2 fixation but often ignored microbes are also involved in CO2 fixation by different CO2 fixation pathways. Since desert represents sparse green cover, we cannot ignore the microbes present in such soil, which would be contributing for carbon fixation. Oligotrophic soil like desert harbours microbes that are capable of surviving in low-carbon conditions, thus opening an area of research for the study of CO2 fixation by oligotrophs. Since microbes require carbon for their growth, and hence under carbon starvation, they might have the mechanism for autotrophy. By exploring CO2 fixation through oligotrophs of desert, we could probably enrich the desert soil by carbon in near future. Such microbe-based CO2 fixation studies could help us to channelize CO2 for the synthesis of some important bioproducts.

Cefuroxime is a semi-synthetic antibiotic of the cephalosporin group and has broad-spectrum antimicrobial activity. HPLC methods, detailed in various pharmacopoeias, are generally used for the quantitative determination of the potency of cefuroxime but are unable to determine its bioactivity against microorganisms. In comparison to HPLC methods bioassay methods are simple and inexpensive however, a bioassay for the determination of both the potency and bioactivity of cefuroxime has not yet been reported in any pharmacopoeia. This article is focused on the development of a bioassay method for the quantification of cefuroxime axetil in pharmaceutical preparations. The bioassay was performed with different microbial strains and Kocuria rhizophila ATCC-9341 was selected as the most susceptible microorganism against cefuroxime axetil. The percent potency of market samples were estimated by a two-level bioassay method. The potency of market samples of Biocef, Ceftech and Ceftum were found to be 100.71% (RSD 1.09%), 101.07% (RSD 1.16%) and 100.44% (RSD 1.08%), respectively. The Biocef intermediate precision RSD between days and between analysts were 1.08% and 1.01%, respectively. The specificity of the bioassay for the analysis of Biocef was also studied in parallel with the Pharmacopoeial HPLC method and the potency was calculated as 101.09%. Results show that a bioassay method can be used for the quality control of cefuroxime axetil in raw materials and pharmaceutical preparations.

Wide range of dyes and dyestuffs used in textile manufacturing are xenobiotic compounds and attract stricter to strict environmental regulations. The ability of microbial consortia to decolorize and metabolize dyes has long been known, and the use of bioremediation based technologies for treating textile wastewater has attracted interest. These dyes are decolorized by microbial consortia but technologies for their complete mineralization are still not developed. The most logical concept for the removal of azo dyes in biological wastewater treatment systems is based on anaerobic treatment, for the reductive decolorization, in combination with aerobic treatment, for the degradation of the by-products (aromatic amines) generated in the anaerobic bioreactor. Several research and review articles were published on anaerobic decolorization; however, research on complete mineralization of dyes through sequential anaerobic–aerobic bioreactors has received greater attention recently. Bioremediation through sequential anaerobic–aerobic bioreactor system has been reviewed in this article with critical appraisal using data generated through our experiments. While reviewing this work, we realized the importance of microbial diversity in a treatment unit to better understand the functional status to enhance the mineralization activity of the bioreactor.

Dye degradation has gained attention of late due to indiscriminate disposal from user industries. Enhancing efficiency of biological treatment provides a cheaper alternative vis-à-vis other advanced technologies. Dye molecules are metabolized biologically via anoxic and oxic treatments. In this study, bacterial community surviving on dye effluent working in anoxic-oxic bioreactor was analyzed using 16S rDNA approach. Azo-dye decolorizing and degrading bacterial community was enriched in lab-scale two-stage anoxic-oxic bioreactor. 16S rDNA metagenomic libraries of enriched population were constructed, screened and phylogenetically analyzed separately. Removal of approximately 35% COD with complete decolorization was observed in anoxic bioreactor. Process was carried out by uncultured gamma proteobacterium constituting 48% of the total population and 12% clones having homology to Klebsiella. Aromatic amines generated during partial treatment under anoxic bioreactor were treated by aerobic population having 72% unculturable unidentified bacterium and rest of the population consisting of Thauera sp., Pseudoxanthomonas sp., Desulfomicrobium sp., Ottowia sp., Acidovorax sp., and Bacteriodetes bacterium sp.

The laboratory-isolated strains Pseudomonas aeruginosa , Rhodobacter sphaeroides , Proteus mirabilis , Bacillus circulance , NAD 1 and NAD 6 were observed to be predominant in the bacterial consortium responsible for effective decolorization of the azo dyes. The kinetic characteristics of azo dye decolorization by bacterial consortium were determined quantitatively using reactive vinyl sulfonated diazo dye, remazol black-B (RB-B) as a model substrate. Effects of substrate (RB-B) concentration as well as different substrates (azo dyes), environmental parameters (temperature and pH), glucose and other electron donor/co-substrate on the rate of decolorization were investigated to reveal the key factor that determines the performance of dye decolorization. The activation energy ( E a ) and frequency factor ( K 0 ) based on the Arrhenius equation was calculated as 11.67 kcal mol −1 and 1.57 × 10 7 mg l g MLSS −1 h −1 , respectively. The Double-reciprocal or Lineweaver–Burk plot was used to evaluate V max , 15.97 h −1 and K m , 85.66 mg l −1 . The two-stage anoxic–oxic reactor system has proved to be successful in achieving significant decolorization and degradation of azo dyes by specific developed bacterial consortium with a removal of 84% color and 80% COD for real textile effluents vis-a-vis ≥90% color and COD removal for synthetic dye solution.

In this work, the coagulation-flocculation process was used as pretreatment for purified terephthalic acid (PTA) wastewater with the objective of improving its overall biodegradability. PTA production generates wastewaters with toxicants p-xylene [1,4-dimethyl-benzene (C8H10)], a major raw material used in the production process, along with some of the intermediates, viz., p-toluic acid, benzoic acid, 4-carboxybenzaldehyde, phthalic acid and terephthalic acid. These compounds affect the bio-oxidation process of wastewater treatment; hence removal of these constituents is necessary, prior to conventional aerobic treatment. This paper addresses the application of coagulation-flocculation process using chemical coagulants, viz., aluminium sulphate (alum), polyaluminium chloride (PAC), ferrous sulphate and ferric chloride in combination with anionic polyelectrolyte. Polyaluminium chloride (PAC) in conjunction with lime and polyelectrolyte removed about 63.1% chemical oxygen demand (COD) and 45.2% biochemical oxygen demand (BOD) from PTA wastewater. Coagulation-flocculation process coupled with aerobic bio-oxidation treatment of PTA wastewater achieved, COD & BOD removals of 97.4% and 99.4%, respectively. The biodegradability enhancement evaluated in terms of the BOD5/COD ratio, increased from 0.45 to 0.67 at the optimum conditions. The results obtained from these studies indicate that the coagulation-flocculation process could be a suitable pretreatment method in reducing toxicity of PTA wastewater whilst enhancing biodegradability for aerobic biological treatment scheme.

Coliphages represent a process indicator for fecal pollution. The coal bed concentration method prepared for enterovirus was refined for a concentration of coliphages. A bed made from 1.5 g of 120-mesh coal powder was used for concentrating coliphage from 200 mL of a water sample with or without the addition of aluminum chloride at different pH values. The isolated E. coli strain EC-R8 was found to be more susceptible to the desired coliphage and showed significant coliphage-coliform response, with clear plaque used for further studies. The complete coliphage adsorption was achieved with the addition of 0.0005 M AlCl 3 at pH 6.0. Adsorbed coliphages were eluted with 3% beef extract in Mcllvaine buffer at pH 7.1, with an average recovery of 78.74%. This concentration technique was applied for the detection of coliphages from the well water of Nagpur city (India) and found to contain coliphages in the range of 2 to 28 plaque-forming units per liter (PFU/L).