Genomic analysis of acid tolerance genes and deciphering the function of ydaG gene in mitigating acid tolerance in Priestia megaterium.

Adverse environmental conditions, such as acid stress, induce bacteria to employ several strategies to overcome these stressors. These strategies include forming biofilms and activating specific molecular pathways, such as the general stress response (GSR). The genome of Priestia megaterium strain G18 was sequenced using the Illumina NextSeq 500 system, resulting in a de novo assembly of 80 scaffolds. The scaffolded genome comprises 5,367,956 bp with a GC content of 37.89%, and was compared to related strains using the MiGA web server, revealing high similarity to P. megaterium NBRC 15308 and P. aryabhattai B8W22 with ANI scores of 95.4%. Phylogenetic and ribosomal multilocus sequence typing (rMLST) analyses, based on the 16S rRNA and ribosomal protein-encoding alleles, confirmed close relationships within the P. megaterium species. Functional annotation identified 5,484 protein-coding genes, with 72.31% classified into 22 COG categories, highlighting roles in amino acid transport, transcription, carbohydrate metabolism, and ribosomal structure. An in-depth genome analysis of P. megaterium G18 revealed several key genes associated with acid tolerance. Targeted inactivation of the ydaG gene from SigB regulon, a general stress response gene, significantly reduced growth under acidic conditions compared to the wild type. qRT-PCR analysis showed increased ydaG expression in acidic conditions, further supporting its role in acid stress response. Microscopic analysis revealed no morphological differences between wild-type and mutant cells, suggesting that ydaG is not involved in maintaining cellular morphology but in facilitating acid tolerance through stress protein production. This research contributes to understanding the molecular mechanisms underlying acid tolerance in soil bacteria, P. megaterium, shedding light on potential applications in agriculture and industry.

Fermented Polyherbal Formulation Ameliorates the Severity of Acute Multiple Antibiotics-Resistant Pseudomonas aeruginosa-Infected Burn Wound in a Rat Burn Model.

Pseudomonas aeruginosa, a Gram-negative opportunistic bacterium, has emerged as a cause of life-threatening infections in burn wounds. Current therapeutic approaches through wound dressings and systemic medicines are far from satisfactory; resistance to more than two antibiotics shown by pathogens contributes to failures of therapy causing mortality. This animal study was conducted to check the efficacy of one Ayurveda-based fermented polyherbal preparation (AP 01) against multiple antibiotics-resistant (MAR) P. aeruginosa HW01-infected rat burn wounds. AP-01 was applied on artificially infected burn wound on a rat model infected with MAR P. aeruginosa to register the healing effects in terms of reduction in residual wound area percentage, the presence of C-reactive protein in blood, and the presence of viable bacteria colony. Topical application with conventional antibiotics served as a positive control. The polyherbal preparation had reduced the infected residual burn wound area at 40.63% ± 0.69 from the initial burn wound area within two weeks after a single intervention, whereas residual burn wound area remained much higher in the case of animals left untreated and in the case of the animals treated with control drug. Restoration to the normalcy of serum C-reactive protein level was also achieved earlier in the case of polyherbal AP-01-treated groups than in other groups. Fermented formulations using components of AP-01 singly or in different combinations had never been tested earlier for topical application in infected burn wound. The formulation of AP-01 was found superior in terms of the rate of healing and control of infection by MAR P. aeruginosa strains in burn wounds in rat models.

What transcriptomics and proteomics can tell us about a high borate perturbed boron tolerant Bacilli strain.

A variety of genes work together to allow the bacterium Lysinibacillus sp. OL1 to survive and grow under B-stress circumstances. This bacterium was previously identified and described from agricultural soil treated with a boron fertilizer. The effects of B-stress on OL1 cells cultured in the presence of 200 mM boric acid were evaluated as changes in the log-phase cell transcriptome and proteome. OL1 has been found to upregulate all genes involved in producing critical macromolecules when exposed to B-stress. It was also observed that genes governing energy supply lines were in higher expression stages, indicating that they were more likely to support the increased production of macromolecules and stress-induced proteins, such as efflux proteins, to reduce boron damage and prevent boron accumulation inside the cell. It has been explained how the hub genes and bottleneck genes cooperate to survive boron stress and support bacterial growth. The proteome results have significantly confirmed the boron tolerance paradigm. Thus, the current study has improved our understanding of the bacterial B-stress response mechanism and opened new research directions.

Draft Genome Sequences from Two Gram-Negative Bacteria, Serratia sp. Strain EWG9 and Leclercia sp. Strain EMC7, Isolated from the Earthworm Eisenia fetida.

We present the draft genome sequences of two bacterial strains that are putatively unique species and belong to two different Gram-negative genera: Serratia sp. EWG9 and Leclercia sp. EMC7, recovered from the gut and cast, respectively, of the compost worm Eisenia fetida.

Inducible boron resistance via active efflux in Lysinibacillus and Enterococcus isolates from boron-contaminated agricultural soil.

Phylogenetically diverse bacteria tolerate high boron concentrations while others require it for metabolic purposes despite the metalloid being toxic beyond a threshold. Boron resistance and plant growth promoting attributes of two bacterial strains, Lysinibacillus sp. OL1 and a novel Enterococcus sp. OL5, isolated from boron-fertilizer-amended cauliflower fields were investigated in this study. OL1 and OL5 grew efficiently in the presence of 210-230 mM boron, and resistance was found to be inducible by small amounts of the element: 5 to 50 mM boron pre-exposure progressively shortened the lag phase of growth in the presence of 200 mM boron. Intracellular boron accumulation was also found to be regulated by the level of pre-exposure: no induction or induction by small amounts led to higher levels of intracellular accumulation, whereas induction by high concentrations led to lower accumulation. These data, in the context of the strains' overall resistance towards 200 mM boron, indicated that induction by higher boron concentrations turned potential efflux mechanisms on, while resistance was eventually achieved by continuous cellular entry and exit of the ions. Involvement of solute efflux in boron resistance was corroborated by the genome content of the isolates (genes encoding proteins of the ATP-binding cassette, major facilitator, small multidrug resistance, multi antimicrobial extrusion, and resistance-nodulation-cell division, family/superfamily). Bacteria such as OL1 and OL5, which resist boron via influx-efflux, potentially lower boron bioavailability, and therefore toxicity, for the soil microbiota at large. These bacteria, by virtue of their plant-growth-promoting attributes, can also be used as biofertilizers.

Draft Genome Sequences of Four Novel Strains of Microbes Isolated from Lepidocephalichthys guntea.

We report the draft genome sequences of four bacterial strains (all of which are putatively novel species) belonging to four different genera. The Gram-positive Bacillus sp. strain GG161 and Rhodococcus sp. strain GG48 and the Gram-negative Achromobacter sp. strain GG226 and Shigella sp. strain GCP5 were all isolated from the gut of the optionally intestine-breathing freshwater fish Lepidocephalichthys guntea.

Functional characterization of thermotolerant microbial consortium for lignocellulolytic enzymes with central role of Firmicutes in rice straw depolymerization.

Rice (Oryza sativa L.) straw, an agricultural waste of high yield, is a sustainable source of fermentable sugars for biofuel and other chemicals. However, it shows recalcitrance to microbial catalysed depolymerization. We herein describe development of thermotolerant microbial consortium (RSV) from vermicompost with ability to degrade rice straw and analysis of its metagenome for bacterial diversity, and lignocellulolytic carbohydrate active enzymes (CAZymes) and their phylogenetic affiliations. RSV secretome exhibited cellulases and hemicellulases with higher activity at 60 °C. It catalysed depolymerization of chemical pretreated rice straw as revealed by scanning electron microscopy and saccharification yield of 460 mg g-1 rice straw. Microbial diversity of RSV was distinct from other compost habitats, with predominance of members of phyla Firmicutes, Proteobacteria and Bacteroidetes; and Pseudoclostridium, Thermoanaerobacterium, Chelatococcus and Algoriphagus being most abundant genera. RSV harboured 1389 CAZyme encoding ORFs of glycoside hydrolase, carbohydrate esterase, glycosyl transferase, carbohydrate binding module and auxiliary activity functions. Microorganisms of Firmicutes showed central role in lignocellulose deconstruction with importance in hemicellulose degradation; whereas representatives of Proteobacteria and Bacteroidetes contributed to cellulose and lignin degradation, respectively. RSV consortium could be a resource for mining thermotolerant cellulolytic bacteria or enzymes and studying their synergism in deconstruction of chemically pretreated rice straw.

Draft Genome Sequences of Two Boron-Tolerant, Arsenic-Resistant, Gram-Positive Bacterial Strains, Lysinibacillus sp. OL1 and Enterococcus sp. OL5, Isolated from Boron-Fortified Cauliflower-Growing Field Soils of Northern West Bengal, India.

Two novel boron-tolerant, arsenic-resistant, Gram-positive bacterial strains, Lysinibacillus sp. OL1 and Enterococcus sp. OL5, were isolated from boron fertilizer-amended cauliflower plantation field soils in India. Here, we report the draft genome sequences of OL1 (4.87 Mb) and OL5 (3.93 Mb) to explore the intricacies of boron tolerance in bacteria.

The antioxidant rich active principles of Clerodendrum sp. controls haloalkane xenobiotic induced hepatic damage in murine model.

Clerodendrum is a plant with potent antioxidant activity and has been frequently employed as a traditional remedy against bronchitis, asthma, liver and stomach disorders. Three species of genus Clerodendrum namely Clerodendrum indicum, C. colebrookianum and C. inerme (Syn. Volkameria inermis) were investigated for their possible activity against oxidative stress induced liver injury. Apart from generation of Reactive Oxygen Species (ROS) in the WRL-68 cell line (human hepatic cell line), in-vitro and in-vivo antioxidant assays were also assessed. Features of immune cell proliferation (MTT) were analyzed thoroughly. Gas Chromatography-Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FTIR) analyses have been performed to identify the active biological compounds. These active biological compounds were further subjected to molecular docking. The antioxidant activity of three Clerodendrum sp. was significantly high in DPPH, nitric oxide, hydroxyl radical and hydrogen peroxide etc. Biochemical parameters like catalase, superoxide dismutase (SOD) and reduced glutathione (GSH) were generated in excess due to CCl4 administration, which was ameliorated by treating with Clerodendrum extract. The phytochemical 24,25-Dihydroxyvitamin D shows excellent binding affinity in Autodock Vina. The present study provided convincing evidences that C. indicum and C. inerme showed good result but C. colebrookianum performed better by almost all means.

Amelioration of CCl4 induced liver injury in swiss albino mice by antioxidant rich leaf extract of Croton bonplandianus Baill.

The progress in industrialization has blessed mankind with a technologically superior lifestyle but poor management of industrial waste has in turn poisoned nature. One such chemical is carbon tetra chloride (CCl4), which is a potent environmental toxin emitted from chemical industries and its presence in the atmosphere is increasing at an alarming rate. Presence of CCl4 in human body is reported to cause liver damage through free radical mediated inflammatory processes. Kupffer cells present in the liver are potentially more sensitive to oxidative stress than hepatocytes. Kuffer cells produced tumor necrosis factor-α (TNF-α) in response to reactive oxygen species (ROS), that might further cause inflammation or apoptosis. In this study hepatoprotective capacity of antioxidant rich extract of Croton bonplandianus Baill. (CBL) was evaluated on CCl4 induced acute hepatotoxicity in murine model. Hydro-methanolic extract of C. bonplandianus leaf was used for evaluation of free radical scavenging activity. Liver cells of experimental mice were damaged using CCl4 and subsequently hepatoprotective potential of the plant extract was evaluated using series of in-vivo and in-vitro studies. In the hepatoprotective study, silymarin was used as a positive control. Antioxidant enzymes, pro-inflammatory markers, liver enzymatic and biochemical parameters were studied to evaluate hepatoprotective activity of Croton bonplandianus leaf extract. Free radical scavenging activity of CBL extract was also observed in WRL-68 cell line. The phytochemicals identified by GCMS analysis were scrutinized using in-silico molecular docking procedure. The results showed that CBL extract have potent free radical scavenging capacity. The biochemical parameters were over expressed due to CCl4 administration, which were significantly normalized by CBL extract treatment. This finding was also supported by histopathological evidences showing less hepatocellularnecrosis, inflammation and fibrosis in CBL and silymarin treated group, compared to CCl4 group. ROS generated due to H2O2 in WRL-68 cell line were normalize in the highest group (200 µg/ml) when compared with control and negative control (CCl4) group. After molecular docking analysis, it was observed that the compound α-amyrin present in the leaf extract of C. bonplandianus has better potentiality to protect hepatocellular damages than the standard drug Silymarin. The present study provided supportive evidence that CBL extract possesses potent hepatoprotective capacity by ameliorating haloalkane induced liver injury in the murine model. The antioxidant and anti-inflammatory activities also affirm the same. The synergistic effects of the phytochemicals present in CBL are to be credited for all the hepatoprotective activity claimed above.

Lignin-graft-Polyoxazoline Conjugated Triazole a Novel Anti-Infective Ointment to Control Persistent Inflammation.

Lignin, one of the most abundant renewable feedstock, is used to develop a biocompatible hydrogel as anti-infective ointment. A hydrophilic polyoxazoline chain is grafted through ring opening polymerization, possess homogeneous spherical nanoparticles of 10-15 nm. The copolymer was covalently modified with triazole moiety to fortify the antimicrobial and antibiofilm activities. The hydrogel was capable of down regulating the expression level of IL-1ß in LPS induced macrophage cells, and to cause significant reduction of iNOS production. It supported cellular anti-inflammatory activity which was confirmed with luciferase assay, western blot, and NF-κB analysis. This novel lignin-based hydrogel tested in-vivo has shown the abilities to prevent infection of burn wound, aid healing, and an anti-inflammatory dressing material. The hydrogel reported here provides a new material platform to introduce a cost-effective and efficient ointment option after undertaking further work to look at its use in the area of clinical practice.