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BACKGROUND: The intestine is a barrier resisting various stress responses. Intrauterine growth restriction (IUGR) can cause damage to the intestinal barrier via destroying the balance of intestinal epithelial cells' proliferation and apoptosis. Bacillus subtilis has been reported to regulate intestinal epithelial cells' proliferation and apoptosis. Thus, the purpose of this study was to determine if B. subtilis could regulate intestinal epithelial cells' proliferation and apoptosis in intrauterine growth restriction suckling piglets. RESULTS: Compared with the normal birth weight group, the IUGR group showed greater mean optical density values of Ki‐67‐positive cells in the ileal crypt (P < 0.05). IUGR resulted in higher ability of proliferation and apoptosis of intestinal epithelial cells, by upregulation of the messenger RNA (mRNA) or proteins expression of leucine rich repeat containing G protein coupled receptor 5, Caspase‐3, Caspase‐7, β‐catenin, cyclinD1, B‐cell lymphoma‐2 associated agonist of cell death, and BCL2 associated X (P < 0.05), and downregulation of the mRNA or protein expression of B‐cell lymphoma‐2 and B‐cell lymphoma‐2‐like 1 (P < 0.05). However, B. subtilis supplementation decreased the mRNA or proteins expression of leucine rich repeat containing G protein coupled receptor 5, SPARC related modular calcium binding 2, tumor necrosis factor receptor superfamily member 19, cyclinD1, Caspase‐7, β‐catenin, B‐cell lymphoma‐2 associated agonist of cell death, and Caspase‐3 (P < 0.05), and increased the mRNA expression of B‐cell lymphoma‐2 (P < 0.05). CONCLUSION: IUGR led to excessive apoptosis of intestinal epithelial cells, which induced compensatory proliferation. However, B. subtilis treatment prevented intestinal epithelial cells of IUGR suckling piglets from excessive apoptosis. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

A new series of N-benzyl-2,5-dihydro-1H-pyrrole-linked benzopyrimidines 5 and 6 were synthesized via the 1,3-dipolar cycloaddition reaction of acetylenic dipolarophiles 3 and 4 with azomethine ylide generated in situ from N-(methoxymethyl)-N-(trimethylsilyl)benzylamine and evaluated for their in vitro antimicrobial activity. The structures of the prepared products were characterized using 1H/13C NMR, IR, and ESI-HRMS techniques. Further, the synthesized compounds were assessed for their antibacterial activity against five types of pathogenic bacteria and three fungal strains. The results show that the degree of antimicrobial effect displayed by the synthesized compounds was variable. Notably, compounds 2a, 2d, 2f, 3e, 3h, 4a, 4d, 4f, 5a, 5b, 5d, 5f, 6b, and 6c showed high activity against the bacterial strains, with MIC values ranging from 15.12 to 15.62 μg mL−1. Moreover, compounds 2a, 2f, 5a, 5b, 5d, 5g, and 5h exhibited promising antifungal activity against three Candida strains, with MIC values ranging from 62.5 to 125 μg mL−1. In addition, molecular docking studies performed to identify the most effective antibacterial compounds against B. subtilis highlighted the high binding affinity of the synthesized compounds toward Bacillus subtilis YdiB (pdb: 5MVR). [ABSTRACT FROM AUTHOR]

Background Aims Methods Results Conclusions Skin aging is one of the most abundant aging‐related disorders that can be accelerated by excessive exposure to ultraviolet irradiation. Topically applied fermented skincare ingredients have gained mounting attentions due to their high concentration of various skin nourishing nutrients and bioactive components and low skin irritation potency.In the present study, we aim to fully demonstrate the skin‐related benefits of a novel extract of Thermus thermophilus and Bacillus subtilis mixed‐culture ferment (TBFE).TBFE was prepared through an innovative mixed‐culture fermentation process. The contents of nutrients and bioactive ingredients were quantified by different methods accordingly. Both in vitro tests and randomized controlled human trial were utilized to further demonstrate multifaceted beneficial effects on human skin, as well as the potential mechanisms.Our results showed that TBFE upregulated the expression of type IV collagen, elastin, aquaporin‐3, and dermal‐epidermal junction markers, while inhibited production of melanin, in different skin cell models. Moreover, TBFE inhibited the generation of reactive oxygen species and pro‐inflammatory mediators induced by ultraviolet irradiation in normal human keratinocytes, while stimulated autophagy in senescent keratinocytes. Results from clinical studies confirmed those in vitro findings, demonstrating that TBFE at 5% and 20% concentration provides anti‐aging properties in subjects with sensitive skin, in terms of improving wrinkles, moisturization, and skin lightening.In summary, we demonstrate that a novel mixed‐culture ferment extract has promising anti‐aging effects, which may be attributed to anti‐oxidation, anti‐inflammation, and promotion of autophagy in skin cells. [ABSTRACT FROM AUTHOR]

ABSTRACT In our work, we have successfully synthesized twelve diorganotin(IV) complexes (4–15) from different hydrazone ligands and diorganotin(IV) dichlorides. The complexes have a general formula [R2SnL1–3], where R = Et, Me, Bu, and Ph groups. The hydrazone ligands (1–3) were synthesized through a condensation reaction involving 3‐ethoxysalicylaldehyde with different benzhydrazide derivatives. All the synthesized hydrazone ligands and their complexes underwent screening by employing numerous spectroscopic and physicochemical techniques, such as molar conductance measurements, mass spectrometry, (1H, 119Sn, and 13C) NMR, SEM‐EDAX, and FT‐IR. Spectroscopic analysis revealed that the hydrazone ligands were attached to tin atoms in a tridentate fashion via ONO donor atoms, suggesting a pentacoordinated geometry for the complexes. Furthermore, the X‐ray crystallography analysis for Complex 6 revealed the distorted square pyramidal geometry around the tin atom. Moreover, a DFT study was also carried out for Ligand 3 and its complexes (12–15) by employing B3LYP/LanL2DZ theory level (Gaussian 9 software package) to obtain the optimized geometry and global reactivity descriptors, structural behavior, and their efficacy against different microbes. To assess the biological efficacy of synthesized compounds, an in vitro antimicrobial assay was conducted against different fungal (Aspergillus niger and Candida albicans) and bacterial (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa) strains. The results of the antimicrobial assay demonstrated that Complexes 7, 11, and 15 depicted better results against E. coli strain and C. albicans strain, implying that the higher lipophilic character of these complexes facilitates their easy passage through the cell membrane of microbes. Furthermore, an ADMET study was carried out to evaluate the toxicity score of synthesized compounds. [ABSTRACT FROM AUTHOR]

Deep eutectic solvents (DESs) are gaining rapid prominence in numerous biocatalysis processes due to their green characteristics, biodegradability, low cost, and simple preparation compared to harsh organic solvents and toxic ionic liquids. However, natural enzymes tend to show activity reduction, even inactivation in the presence of many DESs. Here, we present the first rational design approach to achieve enzymes resistant to both DESs and high temperatures. Using the interaction pattern between DESs and BSLA (Bacillus subtilis lipase A, our model enzyme) derived from all-atom molecular dynamics (MD) simulations, we formulated a solvation-guided engineering strategy. This was established after assessing 33 structural, solvation, and energy observables. We rationally designed and experimentally tested 36 single substitutions, of which 28 (a 77.78% success rate) exhibited improvements in at least two DES cosolvents: choline chloride (ChCl) : acetamide, tetrabutylphosphonium bromide (TBPB) : ethylene glycol (EG), and ChCl : EG. Additionally, through stepwise recombination, we identified two robust BSLA variants, D64H/R107L/E171Y and D64H/R142L, showing stability improvements of up to 4.4-fold and 3.2-fold in three DESs and at 50 °C, respectively. Further MD studies demonstrated that (i) the restricted overall structure but fine-tuned local flexibility and (ii) increased water but decreased DES molecules at substituted sites are two main factors contributing to the enhanced multiple DES resistance. Overall, solvation-guided engineering offers an efficient and rational approach for designing lipases tolerant to DES cosolvents and elevated temperatures, with a considerable potential for adaptation to other enzymes. [ABSTRACT FROM AUTHOR]

Maltotetraose amylase (Mta) catalyzes the hydrolysis of amylaceous polysaccharides into maltotetraose, which is an important functional sugar used in the food industry. However, the lack of efficient expression systems for recombinant Mta has hindered its scale-up production and application. In this study, a codon-optimized mta gene from Pseudomonas saccharophila was efficiently produced in Bacillus subtilis by optimizing the regulatory elements. First, a plasmid library containing 173 different signal peptide sequences placed upstream of mta gene was constructed, and transformed into B. subtilis strain WB800N(amyEΔ1) for high-throughput screening. The signal peptide yhcR was found to significantly enhance the secretion of Mta, reaching an activity of 75.4 U/mL in the culture medium. After optimization of the promoters, the Mta activity was further increased to 100.3 U/mL using a dual-promoter PHpaIIPamyE. Finally, the carbon sources and nitrogen sources for recombinant Mta production were optimized, yielding a highest Mta activity of 288.9 U/mL under the optimal culture conditions. The crude enzyme solution containing recombinant Mta produced a highest maltotetraose yield of 70.3% with 200 g/L of maltodextrin as the substrate. Therefore, the present study have demonstrated a high yield of Mta produced in B. subtilis, laying the foundation for large-scale Mta production and application. [ABSTRACT FROM AUTHOR]

Aims: This study evaluated the impact of wheat straw return and microbial agent application on rice field environments. Methods: Using Rice variety Chuankangyou 2115 and a microbial mix of Bacillus subtilis and Trichoderma harzianum. Five treatments were tested: T1 (no straw return), T2 (straw return), T3, T4, and T5 (straw return with varying ratios of Bacillus subtilis and Trichoderma harzianum). Results: Results indicated significant improvements in rice root length, surface area, dry weight, soil nutrients, and enzyme activity across T2-T5 compared to T1, enhancing yield by 3.81-26.63%. T3 (50:50 microbial ratio) was optimal, further increasing root dry weight, soil enzyme activity, effective panicle and spikelet numbers, and yield. Dominant bacteria in T3 included MBNT15, Defluviicoccus, Rokubacteriales, and Latescibacterota. Higher Trichoderma harzianum proportions (75% in T5) increased straw decomposition but slightly inhibited root growth. Correlation analysis revealed a significant positive relationship between yield and soil microorganisms like Gemmatimonadota and Firmicutes at the heading stage. Factors like dry root weight, straw decomposition rate postjointing stage, and elevated soil enzyme activity and nutrient content from tiller to jointing stage contributed to increased panicle and spikelet numbers, boosting yield. Conclusion: The optimal Bacillus subtilis and Trichoderma harzianum ratio for straw return was 50:50, effectively improving soil health and synergizing high rice yield with efficient straw utilization. [ABSTRACT FROM AUTHOR]

Introduction: Toxic heavy metal pollution has been considered a major ecosystem pollution source. Unceasing or rare performance of Pb2+ to the surrounding environment causes damage to the kidney, nervous, and liver systems. Microbial remediation has acquired prominence in recent decades due to its high efficiency, environment-friendliness, and cost-effectiveness. Methods: The lead biosorption by Bacillus subtilis was optimized by two successive paradigms, namely, a definitive screening design (DSD) and an artificial neural network (ANN), to maximize the sorption process. Results: Five physicochemical variables showed a significant influence (p < 0.05) on the Pb2+ biosorption with optimal levels of pH 6.1, temperature 30°C, glucose 1.5%, yeast extract 1.7%, and MgSO4.7H2O 0.2, resulting in a 96.12% removal rate. The Pb2+ biosorption mechanism using B. subtilis biomass was investigated by performing several analyses before and after Pb2+ biosorption. The maximum Pb2+ biosorption capacity of B. subtilis was 61.8 mg/g at a 0.3 g biosorbent dose, pH 6.0, temperature 30°C, and contact time 60 min. Langmuir's isotherm and pseudo-second-order model with R² of 0.991 and 0.999 were suitable for the biosorption data, predicting a monolayer adsorption and chemisorption mechanism, respectively. Discussion: The outcome of the present research seems to be a first attempt to apply intelligence paradigms in the optimization of low-cost Pb2+ biosorption using B. subtilis biomass, justifying their promising application for enhancing the removal efficiency of heavy metal ions using biosorbents from contaminated aqueous systems. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this study, green synthesis of Ag doped ZnO (AZNPR) has been carried out using the sol-gel method at pH 8.0, using two types of microbial cells, namely Aspergillus niger and Bacillus subtilis as capping and reducing agents. This work centred on the phytomediated biosynthesis and characterisation of AZNPR and its biological utility. Based on infrared analysis, phytochemicals present in microbial cells are extracellular enzymes namely proteolytic, amylolytic, lipolytic, cellulolytic, and pectinase, which can be extracted from A. niger and B. subtilis and are responsible for controlling the growth of AZNPR. The formation of AZNPR was characterised using different analyses. The FTIR shows absorption band located at ∼864 cm−1 that shows the interaction between Ag and ZnO. The XRD results showed the presence of crystalline AZNPR in the forms of hexagonal wurtzite ZnO and Ag. Microstructural and size examination showed that AZNPR was in the form of nanorods with homogeneous distribution with a size of rods in A. nige r media 137.0 nm, while in B. subtili s media 197.4 nm. The Ag–ZnO nanorods revealed the ability to inhibit Candida albicans species, a fungus causing infectious disease, with an inhibition zone measuring 2.1–3.2 mm at 24 h. [ABSTRACT FROM AUTHOR]

Bacterial interactions are vital for adapting to changing environments, with quorum sensing (QS) systems playing a central role in coordinating behaviors through small signaling molecules. The RRNPPA family is the prevalent QS systems in Bacillota and mediating communication through secreted oligopeptides, which are processed into active pheromones by extracellular proteases. Notably, in several cases the propeptides show the presence of multiple putative pheromones within their sequences, which has been proposed as a mechanism to diversify peptide-receptor specificity and potentially facilitate new functions. However, neither the processes governing the maturation of propeptides containing multiple pheromones, nor their functional significance has been evaluated. Here, using 2 Rap systems from bacteriophages infecting Bacillus subtilis that exhibit different types of pheromone duplication in their propeptides, we investigate the maturation process and the molecular and functional activities of the produced pheromones. Our results reveal that distinct maturation processes generate multiple mature pheromones, which bind to receptors with varying affinities but produce identical structural and biological responses. These findings add additional layers in the complexity of QS communication and regulation, opening new possibilities for microbial social behaviors, highlighting the intricate nature of bacterial interactions and adaptation. Some bacterial quorum sensing (QS) systems use oligopeptide pheromones to orchestrate adaptive responses to the environment. This study shows that extracellular proteases yield multiple pheromones from tandemly duplicated propeptides, adding complexity to QS responses. [ABSTRACT FROM AUTHOR]

Background: Cymbopogon is a member of the family Poaceae and has been explored for its phytochemicals and bioactivities. Although the antimicrobial activities of Cymbopogon spp. extracts have been extensively studied, comprehensive analyses are required to identify promising compounds for the treatment of antimicrobial resistance. Therefore, this study investigated the antioxidant and antimicrobial properties of Cymbopogon spp. ethanolic extracts in every single organ. Methods: Ethanolic extracts were obtained from three Indonesian commercial species of Cymbopogon spp., namely Cymbopogon citratus (L.) Rendle, Cymbopogon nardus (DC.) Spatf., and Cymbopogon winterianus Jowitt. The leaf, stem, and root extracts were evaluated via metabolite profiling using gas chromatography-mass spectrometry (GC–MS). In silico and in vitro analyses were used to evaluate the antioxidant and antimicrobial properties of the Cymbopogon spp. ethanolic extracts. In addition, bioactivity was measured using cytotoxicity assays. Antioxidant assays were performed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid (ABTS) to determine toxicity to Huh7it-1 cells using a tetrazolium bromide (MTT) assay. Finally, the antimicrobial activity of these extracts was evaluated against Candida albicans, Bacillus subtilis, Staphylococcus aureus, and Escherichia coli using a well diffusion assay. Results: GC–MS analysis revealed 53 metabolites. Of these, 2,5-bis(1,1-dimethylethyl)- phenol (27.87%), alpha-cadinol (26.76%), and 1,2-dimethoxy-4-(1-propenyl)-benzene (20.56%) were the predominant compounds. C. winterianus and C. nardus leaves exhibited the highest antioxidant activity against DPPH and ABTS, respectively. Contrastingly, the MTT assay showed low cytotoxicity. C. nardus leaf extract exhibited the highest antimicrobial activity against E. coli and S. aureus, whereas C. winterianus stem extract showed the highest activity against B. substilis. Furthermore, computational pathway analysis predicted that antimicrobial activity mechanisms were related to antioxidant activity. Conclusions: These findings demonstrate that the leaves had strong antioxidant activity, whereas both the leaves and stems showed great antimicrobial activity. Furthermore, all Cymbopogon spp. ethanolic extracts showed low toxicity. These findings provide a foundation for future studies that assess the clinical safety of Cymbopogon spp. as novel drug candidates. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: To effectively introduce plasmids into Bacillus species and conduct genetic manipulations in Bacillus chassis strains, it is essential to optimize transformation methods. These methods aim to extend the period of competence and enhance the permeability of the cell membrane to facilitate the entry of exogenous DNA. Although various strategies have been explored, few studies have delved into identifying metabolites and pathways associated with enhanced competence. Additionally, derivative Bacillus strains with non-functional restriction-modification systems have demonstrated superior efficiency in transforming exogenous DNA, lacking more explorations in the regulation conducted by the restriction-modification system to transformation process. Results: Transcriptomic comparisons were performed to discover the competence forming mechanism and the regulation pathway conducted by the BsuMI methylation modification group in Bacillus. subtilis 168 under the Spizizen transformation condition, which were speculated to be the preferential selection of carbon sources by the cells and the preference for specific metabolic pathway when utilizing the carbon source. The cells were found to utilize the glycolysis pathway to exploit environmental glucose while reducing the demand for other phosphorylated precursors in this pathway. The weakening of these ATP-substrate competitive metabolic pathways allowed more ATP substrates to be distributed into the auto-phosphorylation of the signal transduction factor ComP during competence formation, thereby increasing the expression level of the key regulatory protein ComK. The expression of ComK upregulated the expression of the negative regulator SacX of starch and sucrose in host cells, reinforcing the preference for glucose as the primary carbon source. The methylation modification group of the primary protein BsuMI in the restriction-modification system was associated with the functional modification of key enzymes in the oxidative phosphorylation pathway. The absence of the BsuMI methylation modification group resulted in a decrease in the expression of subunits of cytochrome oxidase, leading to a weakening of the oxidative phosphorylation pathway, which promoted the glycolytic rate of cells and subsequently improved the distribution of ATP molecules into competence formation. A genetic transformation platform for wild-type Bacillus strains was successfully established based on the constructed strain B. subtilis 168-R−M− without its native restriction-modification system. With this platform, high plasmids transformation efficiencies were achieved with a remarkable 63-fold improvement compared to the control group and an increased universality in Bacillus species was also obtained. Conclusions: The enhanced competence formation mechanism and the regulation pathway conducted by the functional protein BsuMI of the restriction-modification system were concluded, providing a reference for further investigation. An effective transformation platform was established to overcome the obstacles in DNA transformations in wild-type Bacillus strains. [ABSTRACT FROM AUTHOR]

Introduction: Research on Glycyrrhiza uralensis, a nonhalophyte that thrives in saline-alkaline soil and a traditional Chinese medicinal component, is focused on improving its ability to tolerate salt stress to increase its productivity and preserve its "Dao-di" characteristics. Furthermore, the inoculation of bioagents such as Bacillus subtilis to increase plant responses to abiotic stressors is currently a mainstream strategy. Mitogen-activated protein kinase (MAPK), a highly conserved protein kinase, plays a significant role in plant responses to various abiotic stress pathways. Methods: This investigation involved the identification of 21 members of the GuMAPK family from the genome of G. uralensis, with an analysis of their protein conserved domains, gene structures, evolutionary relationships, and phosphorylation sites using bioinformatics tools. Results: Systematic evolutionary analysis of the 21 GuMAPKs classified them into four distinct subgroups, revealing significant differences in gene structure and exon numbers. Collinearity analysis highlighted the crucial role of segmental duplication in expanding the GuMAPK gene family, which is particularly evident in G. uralensis and shows a close phylogenetic relationship with Arabidopsis thaliana, tomato, and cucumber. Additionally, the identification of phosphorylation sites suggests a strong correlation between GuMAPK and various physiological processes, including hormonal responses, stress resistance, and growth and development. Protein interaction analysis further supported the role of GuMAPK proteins in regulating essential downstream genes. Through examination of transcriptome expression patterns, GuMAPK16-2 emerged as a prospective pivotal regulatory factor in the context of salt stress and B. subtilis inoculation, a finding supported by its subcellular localization within the nucleus. Discussion: These discoveries offer compelling evidence for the involvement of GuMAPK in the salt stress response and for the exploration of the mechanisms underlying B. subtilis' enhancement of salt tolerance in G. uralensis. [ABSTRACT FROM AUTHOR]

The hydroxylation of remote C(sp3)−H bonds in aliphatic amino acids yields crucial precursors for the synthesis of high‐value compounds. However, accurate regulation of the regioselectivity of remote C(sp3)−H bonds hydroxylation in aliphatic amino acids continues to be a common challenge in chemosynthesis and biosynthesis. In this study, the Fe(II)/α‐ketoglutarate‐dependent dioxygenase from Bacillus subtilis (BlAH) was mined and found to catalyze hydroxylation at the γ and δ sites of aliphatic amino acids. Crystal structure analysis, molecular dynamics simulations, and quantum chemical calculations revealed that regioselectivity was regulated by the spatial effect of BlAH. Based on these results, the spatial effect of BlAH was reconstructed to stabilize the transition state at the δ site of aliphatic amino acids, thereby successfully reversing the γ site regioselectivity to the δ site. For example, the regioselectivity of L‐Homoleucine (5 a) was reversed from the γ site (1 : 12) to the δ site (>99 : 1). The present study not only expands the toolbox of biocatalysts for the regioselective functionalization of remote C(sp3)−H bonds, but also provides a theoretical guidance for the precision‐driven modification of similarly remote C(sp3)−H bonds in complex molecules. [ABSTRACT FROM AUTHOR]

Although antimicrobial dressings have proven to be crucial in the treatment of wounds, they may give rise to antibiotic‐resistant strains or result in the release of endotoxins after bacterial death, which in turn inhibits wound healing. This study highlights the efficacy of a novel alumina ceramic textile as dressing that utilizes the principles of bacteria capture from the wound bed and inhibition of bacterial infiltration into the wound bed to reduce the bacterial burden and to inhibit the spread of infection, without the involvement of active antimicrobial substances or functional nanoparticles. The alumina textiles are compared to commercial dressings like the non‐woven mesh Cutimed Sorbact and gauzes from LEINA WERKE and performed significantly better in capturing bacteria. They are found to be effective against both Gram‐negative Escherichia coli and Gram‐positive Bacillus subtilis and show promising results in the presence of simulated wound fluid and in artificial wound bed tests from which they can be easily lifted without leaving behind any visible residues. In summary, the alumina textiles exhibit a highly efficient bacterial binding activity, possibly due to the intrinsic material properties of their hierarchical structure including the tricot knit mesh, small fiber diameters, pronounced fiber surface microtopography, and high specific surface area. [ABSTRACT FROM AUTHOR]

Background and Objectives: Colorectal cancer (CRC) is a heterogeneous disease of the colon or rectum arising from adenoma precursors and serrated polyps. Recently, probiotics have been proposed as an effective and potential therapeutic approach for CRC prevention and treatment. Probiotics have been shown to alleviate inflammation by restoring the integrity of the mucosal barrier and impeding cancer progression. Materials and Methods: In this study, we aimed to investigate the immunomodulatory effects of live and UV-killed Bacillus subtilis natto on the inflammatory response in CRC. Caco-2 cells were exposed to various concentrations of live and UVkilled B. subtilis natto, and cell viability was assessed using MTT assay. Gene expression analysis of IL-10, TGF-β, TLR2 and TLR4 was performed using RT-qPCR. Results: Our findings showed that both live and UV-killed B. subtilis natto caused significant reduction in inflammatory response by decreasing the gene expression of TLR2 and TLR4, and enhancing the gene expression of IL-10 and TGF-β in Caco-2 cells as compared to control group. Conclusion: The results of this study suggest that live and UV-killed B. subtilis natto may hold potential as a therapeutic supplement for modulating inflammation in CRC. [ABSTRACT FROM AUTHOR]

In the model organism Bacillus subtilis, a signaling protease produced in the forespore, SpoIVB, is essential for the activation of the sigma factor σK, which is produced in the mother cell as an inactive pro‐protein, pro‐σK. SpoIVB has a second function essential to sporulation, most likely during cortex synthesis. The cortex is composed of peptidoglycan (PG) and is essential for the spore's heat resistance and dormancy. Surprisingly, the genome of the intestinal pathogen Clostridioides difficile, in which σK is produced without a pro‐sequence, encodes two SpoIVB paralogs, SpoIVB1 and SpoIVB2. Here, we show that spoIVB1 is dispensable for sporulation, while a spoIVB2 in‐frame deletion mutant fails to produce heat‐resistant spores. The spoIVB2 mutant enters sporulation, undergoes asymmetric division, and completes engulfment of the forespore by the mother cell but fails to synthesize the spore cortex. We show that SpoIIP, a PG hydrolase and part of the engulfasome, the machinery essential for engulfment, is cleaved by SpoIVB2 into an inactive form. Within the engulfasome, the SpoIIP amidase activity generates the substrates for the SpoIID lytic transglycosylase. Thus, following engulfment completion, the cleavage and inactivation of SpoIIP by SpoIVB2 curtails the engulfasome hydrolytic activity, at a time when synthesis of the spore cortex peptidoglycan begins. SpoIVB2 is also required for normal late gene expression in the forespore by a currently unknown mechanism. Together, these observations suggest a role for SpoIVB2 in coordinating late morphological and gene expression events between the forespore and the mother cell. [ABSTRACT FROM AUTHOR]

Extracellular proteases are a class of public good that support growth of Bacillus subtilis when nutrients are in a polymeric form. Bacillus subtilis biofilm matrix molecules are another class of public good that are needed for biofilm formation and are prone to exploitation. In this study, we investigated the role of extracellular proteases in B. subtilis biofilm formation and explored interactions between different public good producer strains across various conditions. We confirmed that extracellular proteases support biofilm formation even when glutamic acid provides a freely available nitrogen source. Removal of AprE from the NCIB 3610 secretome adversely affects colony biofilm architecture, while sole induction of WprA activity into an otherwise extracellular protease‐free strain is sufficient to promote wrinkle development within the colony biofilm. We found that changing the nutrient source used to support growth affected B. subtilis biofilm structure, hydrophobicity and architecture. We propose that the different phenotypes observed may be due to increased protease dependency for growth when a polymorphic protein presents the sole nitrogen source. We however cannot exclude that the phenotypic changes are due to alternative matrix molecules being made. Co‐culture of biofilm matrix and extracellular protease mutants can rescue biofilm structure, yet reliance on extracellular proteases for growth influences population coexistence dynamics. Our findings highlight the intricate interplay between these two classes of public goods, providing insights into microbial social dynamics during biofilm formation across different ecological niches. [ABSTRACT FROM AUTHOR]

RNase Y is a key endoribonuclease that regulates global mRNA turnover and processing in Bacillus subtilis and likely many other bacteria. This enzyme is anchored to the cell membrane, creating a pseudo-compartmentalization that aligns with its role in initiating the decay of mRNAs primarily translated at the cell periphery. However, the reasons behind and the consequences of RNase Y's membrane attachment remain largely unknown. In our study, we examined a strain expressing wild-type levels of a cytoplasmic form of RNase Y from its chromosomal locus. This strain exhibits a slow-growth phenotype, similar to that of an RNase Y null mutant. Genome-wide data reveal a significant impact on the expression of hundreds of genes. While certain RNA substrates clearly depend on RNase Y's membrane attachment, others do not. We observed no correlation between mRNA stabilization in the mutant strains and the cellular location or function of the encoded proteins. Interestingly, the Y-complex, a specificity factor for RNase Y, also appears also recognize the cytoplasmic form of the enzyme, restoring wild-type levels of the corresponding transcripts. We propose that membrane attachment of RNase Y is crucial for its functional interaction with many coding and non-coding RNAs, limiting the cleavage of specific substrates, and potentially avoiding unfavorable competition with other ribonucleases like RNase J, which shares a similar evolutionarily conserved cleavage specificity. [ABSTRACT FROM AUTHOR]

The objective of this study was to analyze the chemical composition and evaluate the biological capabilities of the essential oils (EOs) extracted from leaves and stems of wild Aeschynomene indica L. plants by the hydrodistillation method. By using GC-FID/MS, fifty-six and fifty-five compounds, representing 95.1 and 97.6% of the essential oils in the leaves and stems, respectively, were characterized. The predominant constituents of A. indica EOs were (E)-caryophyllene, linalool, viridiflorol, phytol, hexadecanoic acid, trans-verbenol, and α-guaiene. The antibacterial and synergistic activities of the EOs were assessed by microdilution and checkerboard assays. The results revealed a potent inhibition and bactericidal activity against Staphylococcus aureus and Bacillus subtilis with MICs of 0.312–0.625 mg/mL. When combined with traditional antibiotics, the essential oils of A. indica possessed excellent synergistic effects against all tested bacteria. Additionally, the EOs of A. indica leaves showed higher antioxidant activity (IC50 = 0.11 ± 0.01 µg/mL) compared to the stem oil (IC50 = 0.19 ± 0.01 µg/mL) using the ABTS radical scavenging assay. The in vitro cytotoxicity of EOs against human cancer cell lines HepG2, MCF-7, A-549, and HCT-116 was examined, and MTT assays showed that the EOs possessed a significant cytotoxic potential against MCF-7 breast cancer cells, with IC50 values of 10.04 ± 1.82 and 15.89 ± 1.66 μg/mL, and a moderate cytotoxic activity against other tested cells. In conclusion, the A. indica EOs could be considered a potential source of pharmacologically active compounds. [ABSTRACT FROM AUTHOR]

The deterioration of the quality of raw liquor caused by the low content of ethyl hexanoate in Nongxiangxing baijiu has become a pervasive problem in the baijiu industry. Therefore, this study attempted to increase the synthesis of ethyl hexanoate by microorganisms with high esterase activity to increase Zaopei fermentation. The results showed that biofortification was a feasible and important way to improve the quality of the raw liquor and increase the ethyl hexanoate content. Adding Bacillus subtilis, Staphylococcus epidermidis, and Millerozyma farinosa for biofortified fermentation disturbed the microbial community structure of Zaopei and increased the abundance of Wickerhamomyces, Saccharomyces, and Thermoascus. The contents of ethyl hexanoate, ethyl valerate, ethyl caprylate, and ethyl heptanoate also increased noticeably in baijiu. The results of E‐nose and sensory analysis tested and verified that the baijiu in the fortified group had better flavor characteristics. [ABSTRACT FROM AUTHOR]

The nitrilase enzymes are able to convert nitriles to their corresponding carboxylic acid. The present study reports the purification and characterisation of nitrilase (EC 3.5.5.1) which is able to convert acrylonitrile to acrylic acid and ammonia from Bacillus subtilis AGAB 2 (GenBank Ascension number- MW857547) in an optimized mineral medium. The purified nitrilase showed maximum enzyme activity of 5874.82 U ml−1 and, specific enzyme activity of 2097.92 U mg−1 with a 9.95-fold purification. The purified nitrilase showed high specificity for acrylonitrile (Vmax-344.82 µM s−1; Kcat-1603.84 s−1; Km-2.8 mM). The characterization showed that purified nitrilase is most stable and works best under the pH range of 7–8, and temperature range of 28 ºC—37 ºC temperature. The presence of heavy metal like—Co+2, Cu+2, Fe+2, Hg+2, Mg+2, Mn+2, Ni+2, Ca+2, Pb+2, Ba+2 and Zn+2 negatively affected the enzyme activity. Among the tested cosolvents (Methanol, Ethanol, Hexane, DMSO, IPA, Acetone, Ethyl Acetate, Benzene, Chloroform, and Butanol), only Methanol negatively affected nitrilase activity. Hydroxylamine competitively inhibited nitrilase against acrylonitrile, with an IC50 value of 5.5 mM. The biotransformation of acrylonitrile to acrylic acid achieved 95% yield with high purity and selectivity in 48 h, confirmed by HPLC and GCMS analysis. [ABSTRACT FROM AUTHOR]

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are of huge veterinary importance, mainly as vectors of pathogens, such as Bluetongue virus. Currently, there are no effective methods to protect animals against biting midges, as insecticides have limited or short‐lived efficacy. Biting midges are attracted to hosts by carbon dioxide and by their body odours, which are mainly produced by skin bacteria. In humans, it has been shown that differences between individuals in attractiveness to mosquitoes is mediated by these skin bacterial volatiles. This opens the possibility to protect individuals from biting insects by supplementing their skin microbiome with probiotics. In this study, we investigated this approach by culturing sheep skin bacteria on various media and assessing their effects against field‐caught Culicoides (overwhelmingly Obsoletus group species) as well as laboratory‐reared Culicoides nubeculosus (Meigen). Aerococcus urinaeequi, Bacillus safensis, Bacillus subtilis, Jeotgalicoccus psychrophilus, Micrococcus sp. and Staphylococcus equorum were selected to be tested in a dual‐choice Y‐tube olfactometer, assessing their behavioural effects on biting midges. We revealed an avoidance effect on laboratory‐reared C. nubeculosus when testing B. safensis and B. subtilis. Bacillus safensis and Micrococcus sp. yielded significant repellent potential towards field‐caught Culicoides. These two candidates were subsequently tested in a membrane blood‐feeding assay. When the bacterial species B. safensis was applied to the membrane, a feeding reduction of 83% was observed with field‐caught Culicoides. [ABSTRACT FROM AUTHOR]

Survival of probiotics in processed feed can be affected not only by feed processing techniques but also by factors such as storage and gastrointestinal transit of ingested feed. This study investigates the effect of free and spray-dried Bacillus subtilis (BS) in oat β-glucan microcapsules on growth performance, hematology, intestinal microbiota, histology, and immunology of Nile tilapia, Oreochromis niloticus. A total of 400 tilapia were randomly stocked into 25 aquaria in a randomized experimental design and fed one of the following diets: (C) control diet (without synbiotics); (0.1%S) 1g kg−1 diet of unloaded microcapsules and 2.7 × 109 of free BS CFU kg−1 diet; (0.2%S) 2 g kg−1 diet of unloaded microcapsules and 5.4 × 109 of free BS CFU kg−1 diet; (0.1%SM) 1g of microcapsules loaded with 2.7 × 109 BS CFU kg−1 diet; and (0.2%SM) 2g of microcapsules loaded with 5.4 × 109 BS CFU kg−1 diet. After a 60-day feeding trial, fish fed 0.2%SM showed the highest growth performance and best feed utilization compared to fish fed free probiotics and control diet. Intestinal villi were longer, and the submucosa layer was thicker in fish fed both free and microencapsulated probiotics than the control group. Fusobacteriota, Firmicutes, and Bacteroidota were the dominant phyla across all samples accounting for more than 90% of the gut microbiota. No differences were registered in hematological parameters. Phagocytic activity was enhanced in fish fed both 0.2%S and 0.2%SM diets. Microencapsulation has the potential to protect B. subtilis and may constitute a valuable approach for enhancing the viability of probiotics as additives for fish feeds. [ABSTRACT FROM AUTHOR]

The present investigation examined the potential of a previously identified probiotic derived from the intestinal tract of Labeo rohita to modulate the immune system and confer disease resistance when included in a basic diet. The viability of B. subtilis in the feed was assessed at both 4 °C and 28 ± 2 °C, revealing no notable changes. A total of 180 L. rohita fingerlings (19.27 ± 1.96 g) were randomly distributed in four groups (one control and three probiotic-treated groups in triplicate) and fed with the basal (the control) and B. subtilis–supplemented diets (at a concentration of 107, 108, and 109 CFU g−1) over a period of 20 days. Immunological responses, biochemical parameters, and immune-relevant gene expression (interleukin 1β (IL-1β), interferon 1γ (IFN-1γ)) relevant to immunity were analyzed on the 0, 10th, and 20th day post-feeding. In a separate but similar setup, probiotic-fed fish were intraperitoneally challenged with Aeromonas hydrophila (ATCC 7966), and their post-challenge survivability was calculated. After the 20-day feeding period, a significant (P < 0.05) improvement in immune-biochemical response and immune gene expression was evident. Serum glucose and sodium levels exhibited a significant (P < 0.05) decrease. The inclusion of B. subtilis in the diets led to a substantial (P < 0.05) increase in the survival of A. hydrophila–challenged L. rohita, thereby highlighting the potential of B. subtilis COFCAU_BSP3 as a beneficial probiotic for aquaculture. [ABSTRACT FROM AUTHOR]

Plants have limited resources to allocate to defences against infection and herbivory. While interactions between plant responses to microbial and herbivore attack are complex, it is often the case that the induction of one response will act antagonistically to the other. Recent studies have shown that plant growth promoting rhizobacteria, which improves overall plant health and general stress resistance, can enhance both anti-microbial and anti-herbivore defences. We investigated how soil application of the biofungicide Serenade ASO (Bacillus subtilis strain QST 713), which primes plant defences against fungal and bacterial infection and promotes plant growth, affects anti-herbivore defences by measuring both constitutive and induced defences. We applied Serenade one or two times to the soil of tomato plants and measured the numbers of type IV glandular trichomes on leaves, the weight gain of a generalist caterpillar (beet armyworms; BAW), and the activity of two enzymes associated with defence against insects (polyphenoloxidase and peroxidase). Serenade treated plants grew faster and foliage from treated plants had significantly higher numbers of glandular trichomes and higher polyphenoloxidase and peroxidase activities than untreated plants. However, Serenade treatment did not affect the degree of induction of plant defences when damaged by BAW feeding and did not slow the growth rate of BAW relative to plants not treated with Serenade. Therefore, the biofungicide Serenade increased plant growth and altered the densities of trichomes and the activities of two defensive enzymes in plants, but it did not affect overall susceptibility of the plants to a generalist herbivore. [ABSTRACT FROM AUTHOR]

Potato (Solanum tuberosum L.) seed production using the aeroponic system has been widely implemented in Indonesia. However, aeroponic systems in tropical areas such as Indonesia faced obstacles in the form of high wilt attacks caused by the bacteria Ralstonia solanacearum. This research aimed to control R. solanacearum wilt disease in an aeroponic system using various microbes. The research was carried out from September 2022 to January 2023. This research consisted of four stages, namely exploration and isolation of microorganisms, identification of microbial antagonism, testing of in vitro potential microbial antagonism and testing of selected microbial antagonism in the aeroponic system. The nutrients used in the aeroponic system contained R. solanacearum with a concentration of 104 CFU mL-1, while the concentration of the microbial solution used was 108 CFU mL-1. Pseudomonas fluorescence was consistently the best microbe both in vitro and in aeroponics, whereas Bacillus subtilis could only overcome wilt in vitro and could not overcome wilt in aeroponic system. Pseudomonas fluorescence required 5 min soaking time and could increase wilt with increasing soaking time. The consortium between B. subtilis and P. fluorescence with 10 min soaking time could overcome wilt by 85% and increase the number of tubers and tuber weight by 79% and 85%, respectively, so this prospective microbial consortium could be applied to aeroponic systems environmentally friendly in tropical areas with high R. solanacearum opportunities. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a fungal toxin produced by Fusarium that widely occurs in various types of grains and feed. It has strong reproductive toxicity and a wide range of distribution, which will impact food and feed safety. In this study, Bacillus subtilis ZENL09, which is capable of transforming ZEN was investigated. By optimizing the fermentation conditions, the supernatant of Bacillus subtilis ZENL09 fermentation can achieve about 88% degradation rate of ZEN after reacting with ZEN for 12 h. The degradation product was collected by high-performance liquid chromatography, followed by LC-Q-TOF and MS/MS analysis to identify the changes in molecular mass and compound structure. The possible structure of the degradation product was also tentatively determined. Further evaluation of the degradation products' cytotoxicity was conducted using HepG2 cells, and the results indicated a significant reduction in toxicity compared to ZEN. Detoxification experiments on corn steep liquor showed that the fermentation supernatant of Bacillus subtilis degraded 87% of ZEN within 12 h, which further validates its potential application for detoxification in food and agricultural products. [ABSTRACT FROM AUTHOR]

In this research, we employed a microwave-assisted green synthesis method utilizing extracts from Cistus salviifolius L. and Ferula communis L. to synthesize silver nanoparticles (AgNPs). Subsequently, we characterized the biosynthesized AgNPs, both Cs-AgNPs (derived from C. salviifolius) and Fc-AgNPs (derived from F. communis), through UV-Vis, FT-IR, XRD, SEM, and TEM analyses. The AgNPs exhibited a spherical shape with sizes averaging below 11 nm. To assess their antioxidant activity, we employed the DPPH method. Furthermore, we examined the antimicrobial potential of Cs-AgNPs and Fc-AgNPs against pathogenic bacterial strains, including Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli, using the disc diffusion assay and minimum inhibitory concentration. Additionally, we evaluated the cytotoxic effects of Cs-AgNPs and Fc-AgNPs on various cell lines, namely breast adenocarcinoma (MCF-7), colon carcinoma (HT-29), human keratinocytes (HaCaT), and embryonic kidney (HEK-293), employing the Alamar Blue assay. Our findings indicate that the biosynthesized Cs-AgNPs and Fc-AgNPs possess antioxidant, antibacterial, and anticancer activities. Therefore, the current study suggests that Cs-AgNPs and Fc-AgNPs exhibit potential for treating cancer and diseases caused by pathogenic microorganisms. [ABSTRACT FROM AUTHOR]

In this study, bacteria with the best abilities in cellulose degradation, siderophore production, phosphate solubility, and Pythium parasitica inhibition were selected from termite nests and guts. The isolate BTNASP 5-2, BTPK 5-3 and BTNA 5-1 from termite guts exhibited highest in siderophore production index (SPI) (4.16 ± 0.21), phosphate solubilizing index (PSI) (2.10 ± 0.14) and percentage inhibition of radial growth (PIRG) (67.07 ± 4.02 %), respectively. The BGNACMC 4-3 isolated from termite nest gave the highest cellulolytic index (CI) of 5.17 ± 0.24. Bacterial classification was performed using 16s rRNA gene sequencing. The isolates BTPK 5-3, BGNACMC 4-3 and BTNA 5-1 were found closely related to Bacillus cereus, whereas the bacterial isolate BTNASP 5-2 was closely related to Bacillus subtilis. It is also suggested that the Bacillus cereus exhibited a variety of biological activities, denoting the highest cellulase, phosphate-solubilizing and antifungal activities, while Bacillus subtilis produced only a siderophore. The results obtained suggest that the bacteria selected will be used to develop bio-compost to promote plant growth, leading to sustainable farming. [ABSTRACT FROM AUTHOR]

We established confrontations between two different fungi, i.e., the maize anthracnose and stalk rot pathogen Colletotrichum graminicola, and the ubiquitous fungus Aspergillus nidulans, and different biocontrol species, i.e., Bacillus subtilis, Bacillus velezensis, and Bacillus amyloliquefaciens. In all fungus–bacterium confrontations tested, growth arrest and, thus, distance inhibition was observed on solid substrata. LC–MS/MS analyses of culture filtrates suggested formation of several metabolites only synthesized in confrontations. Interestingly, microscopy of fungal hyphae grown in liquid medium showed protrusions and color changes occurred only in media harboring fungus-bacterium confrontations. These observations indicate metabolic re-programming and suggest formation of putative secondary metabolites in interactions involving microbial biocontrol agents. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to investigate the efficacy of Bacillus-based probiotics supplemented at two different levels to modulate the productive performance, egg quality, tibia traits, and specific cecal bacteria counts of Hy-Line Brown layers from 25 to 37 weeks of age. Methods: A total of 216 twenty-five-week-old hens were randomly distributed into 3 experimental diets with 12 replicates of 6 birds per cage. Diets included basal diet supplemented with 0 (CON), 3x108 (PRO1), or 3x109 (PRO2) colony-forming unit (CFU) of the test probiotic containing Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 per kilogram of feed. Results: Improved egg weights and mass at 29 weeks; and feed intake at 31 weeks (p<0.10) were noticed with the probiotic-supplemented PRO1 and PRO2 diets. Considering egg quality, the shell thickness, Haugh units, and yolk color were improved; but yolk cholesterol was lowered (p<0.05) with PRO1 and PRO2 diets at 29 weeks. At both 33 and 37 weeks, the egg-breaking strength, shell color and thickness, albumen height, Haugh units, and yolk color were improved; but yolk cholesterol was similarly lowered (p<0.05) with the PRO1 and PRO2 diets. Improved tibia Ca, ash, weights, and density; and raised cecal counts of Bifidobacteria and Lactobacilli (p<0.05) were noticed with PRO1 and PRO2 diets. Improved tibia P but reduced Clostridia counts (p<0.10) were also observed with the PRO1 and PRO2 diets. Conclusion: Probiotic supplementation of Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 at 3x108 CFU/kg of feed is adequate to significantly improve egg quality, lower yolk cholesterol, enhance several tibia traits, and raise the populations of beneficial cecal bacteria. Modest improvements in several productive parameters and tibia P but reduced Clostridia were also observed; and could warrant further investigation of probiotic effects beyond the current test period. [ABSTRACT FROM AUTHOR]

Clostridium tyrobutyricum strain NRRL B-67062 was previously isolated from an ethanol production facility and shown to produce high yields of butyric acid. In addition, the cell-free supernatant of the fermentation broth from NRRL B-67062 contained antibacterial activity against certain Gram-positive bacteria. To determine the source of this antibacterial activity, we report the genome and genome mining of this strain. The complete genome of NRRL B-67062 showed one circular chromosome of 3,242,608 nucleotides, 3114 predicted coding sequences, 79 RNA genes, and a G+C content of 31.0%. Analyses of the genome data for genes potentially associated with antimicrobial features were sought after by using BAGEL-4 and anti-SMASH databases. Among the leads, a polypeptide of 66 amino acids (PEG 446) contains the DUF4177 domain, which is an uncharacterized highly conserved domain (pfam13783). The cloning and expression of the peg446 gene in Escherichia coli and Bacillus subtilis confirmed the antibacterial property against Lactococcus lactis LM 0230, Limosilactobacillus fermentum 0315-25, and Listeria innocua NRRL B-33088 by gel overlay and well diffusion assays. Molecular modeling suggested that PEG 446 contains one alpha-helix and three anti-parallel short beta-sheets. These results will aid further functional studies and facilitate simultaneously fermentative production of both butyric acid and a putative bacteriocin from agricultural waste and lignocellulosic biomass materials. [ABSTRACT FROM AUTHOR]

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Bacterial illness causes detrimental impacts on fish health and survival and finally economic losses for the aquaculture industry. Antibiotic medication causes microbial resistance, so alternative control strategies should be applied. In this work, we investigated the probiotic-medicated diet as an alternative control approach for antibiotics in treating Vibrio cholerae infection in Nile tilapia (Oreochromis niloticus). One hundred eighty fish (50 ± 2.5 g Mean ± SD) were allocated into six groups in glass aquariums (96 L) in triplicate for 10 days. Groups 1 (G1), G2, and G 3 were intraperitoneally (IP) injected with 0.5 mL sterilized tryptic soy broth and fed on a basal diet, basal diet contained B. subtilis (BS) (1 × 10 5 CFU/ kg-1 diet), and basal diet contained trimethoprim-sulfamethoxazole (TMP-SMX) (1.5 g/kg-1 diet), respectively. Additionally, G4, G5, and G6 were IP challenged with 0.5 mL of V. cholerae (1.5 × 107 CFU) and received the same feeding regime as G 1 to 3, respectively. The results exhibited that the V. cholera-infected fish exhibited skin hemorrhage, fin rot, and the lowest survival (63.33%). Additionally, lowered immune-antioxidant biomarkers (white blood cells count, serum bactericidal activity, phagocytic activity, phagocytic index, and lysozymes) with higher lipid peroxidation marker (malondialdehyde) were consequences of V. cholerae infection. Noteworthy, fish-fed therapeutic diets fortified with BS and TMP-SMX showed a substantial amelioration in the clinical signs and survival. The BS diet significantly improved (P < 0.05) the immune-antioxidant indices of the infected fish compared to the TMP-SMX diet. The current findings supported the use of a BS-enriched diet as an eco-friendly approach for the control of V. cholerae in O. niloticus. [ABSTRACT FROM AUTHOR]

The demand for probiotics is increasing, providing opportunities for food and beverage products to incorporate and market these foods as a source of additional benefits. The most commonly used probiotics belong to the genera of Lactobacillus and Bifidobacterium, and traditionally these bacteria have been incorporated into dairy products, where they have a wider history and can readily survive. More recently, there has been a desire to incorporate probiotics into various food products, including baked goods. In recent years, interest in the use of Bacillus species as probiotics has greatly increased. The spores of various Bacillus species such as Bacillus coagulans and Bacillus subtilis, have significantly improved viability and stability under harsher conditions during heat processing. These characteristics make them very valuable as probiotics. In this review, factors that could affect the stability of Bacillus probiotics in food products are highlighted. Additionally, this review features the existing research and food products that use Bacillus probiotics, as well as future research opportunities. [ABSTRACT FROM AUTHOR]

Fermentation with Bacillus subtilis significantly enhances the physiological activity and bioavailability of soymilk, but the resulting characteristic flavor seriously affects its industrial promotion. The objective of this study was to identify key proteins associated with characteristic flavors in B. subtilis BSNK-5-fermented soymilk using tandem mass tag (TMT) proteomics. The results showed that a total of 765 differentially expressed proteins were identified. Seventy differentially expressed proteins related to characteristic flavor were screened through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. After integrating metabolomics data, fifteen key proteases of characteristic flavor components in BSNK-5-fermented soymilk were further identified, and free ammonia was added. In addition, there were five main formation mechanisms, including the decomposition of urea to produce ammonia; the degradation of glutamate by glutamate dehydrogenase to produce ammonia; the degradation of threonine and non-enzymatic changes to form the derivative 2,5-dimethylpyrazine; the degradation of valine, leucine, and isoleucine to synthesize isovalerate and 2-methylbutyrate; and the metabolism of pyruvate and lactate to synthesize acetate. These results provide a theoretical foundation for the improvement of undesirable flavor in B. subtilis BSNK-5-fermented soy foods. [ABSTRACT FROM AUTHOR]

The fungus Physisporinus vitreus and the bacterium Bacillus subtilis are commonly used organisms in wood biotechnology to increase wood permeability. However, comparative researches on wood destruction by these biological wood decomposers are scarce, particularly from microscopic and chemical perspectives. This study aimed to compare the fungal and bacterial wood destruction patterns over time. Loblolly pine (Pinus taeda L.) heartwood samples were exposed to both organisms for six and nine weeks; microscopic and spectroscopic analyses of the incubated wood were performed, and mass loss of samples was calculated. Both organisms initially destroyed the pit tori and ray parenchyma cells, but B. subtilis was more selective in its consumption of wood components. It exclusively consumed hemicelluloses and free carbohydrates in wood, whereas the fungus destroyed all chemical components simultaneously. The digestion of bordered pits by B. subtilis was more selective than that by P. vitreus, making it a better choice for controlled wood destruction. By extending the exposure duration, the bacterial destruction pattern remained consistent, whereas fungal destruction spread to the secondary wall of the tracheids. Understanding the interactions between microbes and wood can lead to a more effective and efficient use of this natural resource. [ABSTRACT FROM AUTHOR]

The mechanism underlying seed germination promotion by plant growth-promoting rhizobacteria (PGPR) remains unclear. This study investigated the effects of Bacillus subtilis QM3 (a PGPR) and gibberellic acid (GA3) on germination indexes, seedling growth, and amylase activity in wheat seeds (Triticum aestivum L.) (caryopses) at different germination times. The results showed that B. subtilis QM3 exhibited a significantly greater increase in most measured indicators compared to GA3 (p < 0.05), and the coordinated effect of bacteria QM3 and GA3 was superior. GA3 showed no promotion of radicle length (LR) from day 3 to day 7. Conversely, the addition of uniconazole (U), a gibberellin synthesis inhibitor, significantly reduced all indexes except for fresh weight (FW) and root-shoot ratio (RSR). However, both B. subtilis QM3 and GA3 significantly alleviated the inhibition (p < 0.05), especially in the germination rate and amylase activity. The gibberellin (GA) content and expression of two key biosynthetic enzyme genes were also characterized. Further investigations revealed that the GA content in the B. subtilis QM3 group was significantly higher than in the control group at 12 h, 24 h, and 48 h during seed germination (p < 0.05). Moreover, B. subtilis QM3 significantly increased TaGA3ox-B2 expression (p < 0.05), with 1.44 times that of the control group, while there was no significant effect on TaGA20ox-D2 expression. Furthermore, amylase activity was positively correlated with the germination rate (GR) and GA content was positively correlated with amylase activity according to Pearson's correlation analysis, with R2 values of 0.8102 and 0.8365, respectively. Our results demonstrate that one of the mechanisms by which B. subtilis QM3 promotes wheat seed germination may be the induction of GA biosynthesis. [ABSTRACT FROM AUTHOR]

Problem statement: As the intestinal function is intimately affected by fed diets, many kinds of natural substances, prebiotics and probiotics have been supplemented to broilers to increase poultry production by activating intestinal function. The aim of this study was to investigate whether Bacillus subtilis could improve the growth performance and serum biochemistry in broiler chicken. Approach: A total of 200 broiler chicks were randomly assigned to 4 treatment groups, consisting of 2 replicates of 25 birds each. Commercial mash pre-starter, starter and finisher diets were supplemented with 50, 100 and 150 ppm of B. subtilis. Results: Body weight gain was better in all the experimental groups than the control. The growth performance was increased in 150 ppm of B. subtilis. Total cholesterol and Triglycerides were decreased, HDL, Serum Glutamine Pyruvic Transaminase (SGPT), Serum Glutamic Oxaloacetic Transaminase (SGOT) were increased. Conclusion: This study shows that adding B. subtilis to the broiler diet could improve the growth performance, increase feed efficiency and regulate serum biochemical parameters. Based on our study, it could be recommended that addition of B. subtilis at 150 ppm (150 mg/kg) could improve the growth performance of broiler chicken. [ABSTRACT FROM AUTHOR]

Microbial communities such as biofilms are commonly found at interfaces. However, it is unclear how the physical environment of interfaces may contribute to the development and behavior of surface-associated microbial communities. Combining multimode imaging, single-cell tracking, and numerical simulations, here, we found that activity-induced interface bulging promotes colony biofilm formation in Bacillus subtilis swarms presumably via segregation and enrichment of sessile cells in the bulging area. Specifically, the diffusivity of passive particles is ~50% lower inside the bulging area than elsewhere, which enables a diffusion-trapping mechanism for self-assembly and may account for the enrichment of sessile cells. We also uncovered a quasilinear relation between cell speed and surface-packing density that underlies the process of active interface bulging. Guided by the speed-density relation, we demonstrated reversible formation of liquid bulges by manipulating the speed and local density of cells with light. Over the course of development, the active bulges turned into striped biofilm structures, which eventually give rise to a large-scale ridge pattern. Our findings reveal a unique physical mechanism of biofilm formation at air-solid interface, which is pertinent to engineering living materials and directed self-assembly in active fluids. [ABSTRACT FROM AUTHOR]

Endospore-forming bacteria are ubiquitous. Bacterial endospores are multilayered proteinaceous structures that protects the bacterial genome during stress conditions. They are also responsible for a wide range of critical clinical infections in humans. Precise analysis of spore-forming pathogens remains a major challenge in the field of proteomics because spore structures are highly resistant to conventional solubilizers and denaturing agents, such as sodium dodecyl sulfate and urea. We present an ionic liquid-assisted (i-soln) technique of sample preparation, called pTRUST, which enables shotgun analysis of Bacillus subtilis spores even when the starting materials are in the sub-microgram range. In proteomic analysis, this technique shows 50–2000-fold higher sensitivity than other conventional gel-based or gel-free methods (including one-pot sample processing). Using this technique, we identified 445 proteins with high confidence from trace amounts of highly pure spore preparations, including 52 of the 79 proteins (approximately 70%) previously demonstrated to be localized in spores in the SubtiWiki database and detected through direct protein analysis. Consequently, 393 additional proteins were identified as candidates for spore constitutive proteins. Twenty of these newly identified candidates were produced as green fluorescent protein fusion proteins, and each was evaluated for authenticity as a spore constituent using fluorescence microscopy analysis. The pTRUST method's sensitivity and reliability using the i-soln system, together with hitherto unreported proteins in spores, will enable an array of spore research for biological and clinical applications. [ABSTRACT FROM AUTHOR]

DaiDai fruit, a medicinal and edible plant fruit, is abundant in biologically active compounds and has a long history of use in traditional Chinese medicine. This research focuses on utilizing fermentation to develop a functional DaiDai fruit fermentation broth. Lactobacillus, Bacillus subtilis and Saccharomyces cerevisiae were employed in the fermentation process. By conducting screenings of bacterial strains, single factor experiments, and response surface methodology, the total flavonoids, polysaccharides, polyphenols, and 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical scavenging rate were used as the index for selection, ultimately identifying Lactobacillus L-13 as the optimal fermentation strain. The optimal fermentation conditions were determined to be a time of 108 h, a temperature of 43.6°C, and a solid-liquid ratio of 1:15.157 (w/v). Under these conditions, the total flavonoid content reached 412.01 mg/g, representing a 36.71% increase compared to conventional extraction methods. The contents of polysaccharides and polyphenols and the DPPH scavenging rate were also increased. The fermentation broth of DaiDai fruit exhibited inhibitory effects on tyrosinase and melanin production in mouse melanoma cells B16-F10 induced by α-MSH and anti-inflammatory properties in a zebrafish inflammation model. These indicate that the DaiDai fruit fermentation broth possesses antimelanoma, whitening, and anti-inflammatory properties, showcasing significant potential for applications in medicine, cosmetics, and other industries. [ABSTRACT FROM AUTHOR]

AbstractAmylases are highly needful product in food, beverage, and detergent industry. With the objective of isolating α-amylases producing strains, samples were collected from local bakery chimneys and kitchens, Chennai, Tamil Nadu, India. The potential α-amylase producers were isolated and screened by Starch-Iodine plate assay method. The optimized parameters such as 1.5% of starch concentration in the media with 7.2-pH at 35 °C for 36 h enhanced the production of α-amylase. The strain showing high potential, identified by 16s rRNA sequencing was found to be Bacillus subtilis. The Bacillus subtilis strain produced extracellular α-amylase with activity of 1330.14 U/mL. Further partial purification by ammonium sulphate precipitation followed by dialysis and column chromatography enhanced the specific activity to 1695.306 U/mg. The molecular weight of enzymes was found to be around 36 kDa. The enzyme stability and activity characterization revealed that the enzyme was thermotolerant which was able to retain more than 70% activity at 70 °C, 80 °C and 90 °C. The purified enzyme was also found to be Ca2+ independent. The hydrolysis of raw starch from rice, wheat and corn flour using the purified α-amylase from Bacillus subtilis showed significant activity of 72.8%, 76.1% and 65.2% with 4 h incubation respectively. The potentiality of the enzyme as an additive with commercial detergent was also determined to be effective. This study favourably attracts the attention towards industrial applications. [ABSTRACT FROM AUTHOR]

Background: Resistance to antifungal medications poses a significant obstacle in combating fungal infections. The development of novel therapeutics for Candida albicans is necessary due to the increasing resistance of candidiasis to the existing medications. The utilization of biological control is seen as a more advantageous and less hazardous strategy therefore the objective of this study is to identify the antifungal properties of Bacillus subtilis against pathogenic C. albicans. Results: We conducted a study to evaluate the antifungal properties of three bacterial isolates against the human pathogen Candida albicans. One of the bacterial isolates exhibited a potent antifungal activity against this fungal pathogen. This bacterium was identified as Bacillus subtilis based on the 16Sr RNA gene sequence. It exhibited inhibitory efficacy ranging from 33.5 to 44.4% against 15 Candida isolates. The optimal incubation duration for achieving the maximum antifungal activity was determined to be 48 h, resulting in a mean inhibition zone diameter of 29 ± 0.39 mm. The Potato Dextrose agar (PDA) medium was the best medium for the most effective antifungal activity. Incubation temperature of 25oC and medium pH value of 8.0 were the most favorable conditions for maximum antagonistic activity that resulted fungal growth inhibition of 40 ± 0.16 and 36 ± 0.94 mm respectively. Furthermore, the addition of 10.5 mg/ml of bacterial filtrate to C. albicans colonies resulted in 86.51%. decrease in the number of germinated cells. The fungal cell ultrastructural responses due to exposure to B. subtilis filtrate after 48 h were investigated using transmission electron microscopy (TEM). It revealed primary a drastic abnormality that lead to cellular disintegration including folding and lysis of the cell wall, total collapse of the yeast cells, and malformed germ tube following the exposure to the filtrate. However, the control culture treatment had a characteristic morphology of the normal fungal cells featuring a consistently dense central region, a well-organized nucleus, and a cytoplasm containing several components of the endomembrane system. The cells were surrounded by a uniform and intact cell wall. Conclusion: The current study demonstrates a notable antifungal properties of B. subtilis against C. albicans as a result of production of bioactive components of the bacterial exudate. This finding could be a promising natural antifungal agent that could be utilized to combat C. albicans. [ABSTRACT FROM AUTHOR]

The AMP-forming acetyl-CoA synthetase is regulated by lysine acetylation both in bacteria and eukaryotes. However, the underlying mechanism is poorly understood. The Bacillus subtilis acetyltransferase AcuA and the AMP-forming acetyl-CoA synthetase AcsA form an AcuA•AcsA complex, dissociating upon lysine acetylation of AcsA by AcuA. Crystal structures of AcsA from Chloroflexota bacterium in the apo form and in complex with acetyl-adenosine-5′-monophosphate (acetyl-AMP) support the flexible C-terminal domain adopting different conformations. AlphaFold2 predictions suggest binding of AcuA stabilizes AcsA in an undescribed conformation. We show the AcuA•AcsA complex dissociates upon acetyl-coenzyme A (acetyl-CoA) dependent acetylation of AcsA by AcuA. We discover an intrinsic phosphotransacetylase activity enabling AcuA•AcsA generating acetyl-CoA from acetyl-phosphate (AcP) and coenzyme A (CoA) used by AcuA to acetylate and inactivate AcsA. Here, we provide mechanistic insights into the regulation of AMP-forming acetyl-CoA synthetases by lysine acetylation and discover an intrinsic phosphotransacetylase allowing modulation of its activity based on AcP and CoA levels. AMP-forming acetyl-CoA synthetase (AcsA) is an essential enzyme in all domains of life. Here, the authors provide mechanistic insights into regulation of AcsA by lysine acetylation, uncovering a regulatory phosphotransacetylase activity. [ABSTRACT FROM AUTHOR]

In this work, we describe the synthesis of novel Ruthenium (II) complex-based salen Schiff bases. The obtained Ruthenium (II) complexes are characterized using usual spectroscopic and spectrometric techniques, viz., IR, UV-Vis, NMR (1H and 13C), powder X-ray diffraction, and HRMS. Further techniques, such as DTA-TGA and elemental analysis, are used to well establish the structure of the obtained complexes. Octahedral geometries are tentatively proposed for the new Ru(II) complexes. The measured molar conductance for the Ruthenium (II) complexes shows their electrolytic nature (4.24–4.44 S/m). The new Ru(II) complexes are evaluated for their antioxidant and antibacterial activities. The DPPH radical scavenging, FRAP, and total antioxidant capacity (TAC) assays show that the obtained complexes are more potent than the used positive control. They also exhibit promising antibacterial responses against pathogen bacteria: [RuH2L3Cl2] exhibits an important inhibition against Bacillus subtilis DSM 6633, with an inhibition zone of 21 ± 1.41 mm with an MIC value of 0.39 mg/mL, and Proteus mirabilis INH, with 16.50 ± 0.70 mm and an MIC value of 0.78 mg/mL, while [RuH2L2Cl2] exerts interesting antibacterial effects versus Bacillus subtilis DSM 6633 (21 ± 1.41 mm) and Proteus mirabilis INH (25.5 ± 0.70 mm) with equal MIC values of 0.97 mg/mL. [ABSTRACT FROM AUTHOR]

Simple Summary: Bacillus subtilis is a probiotic that can produce surfactin during fermentation. Surfactin is a lipopeptide whose antibacterial activity increases positively with fatty acid chain length, showing great potential to replace antibiotic growth promoters. B. subtilis LYS1 (LYS1) is selected as a strain that can produce high surfactin yields. In addition, previous research has confirmed that surfactin content and LYS1 counts in the fermented product (FP) are positively correlated with weight gain and production efficiency factor in broilers. This study shows that dietary supplementation with 1 to 2.5% LYS1 FP can improve the growth performance, development of intestinal villi, and tibial bone density/mineralization in broilers, and that 1.9% is the optimal supplementation level of LYS1 FP in the diet. This study aimed to investigate the physiochemical characterizations of the fermented product (FP) produced by the high-yield surfactin strain Bacillus subtilis LYS1 (LYS1), as well as its effects on growth performance, carcass traits, intestinal morphology, tibial bone characteristics, and clinical blood biochemistry in broilers. Accordingly, the optimal supplementation of FP for improving growth performance, intestinal villi development, and tibial bone strength in broilers was elucidated using a broken-line quadratic (BLQ) ascending model. Three hundred and sixty 0-day-old Ross 308 broiler chicks, with equal numbers of both sexes, were randomly assigned to dietary supplementation of 2.5% fish meal or 0, 1, 1.5, 2, or 2.5% FP. Each treatment had six replicates, and the experimental period was 5 wk. The LYS1 count, surfactin content, and surfactin composition of the FP were 9.1 log CFU/g, 11.23 mg/g, and C12 to C18, respectively. The FP-supplemented groups improved feed intake, weight gain (WG), and production efficiency factor at 0 to 5 weeks old (p < 0.05) compared with the 0% group. The villus height/crypt depth (V/C) in the jejunum and ileum of the FP-supplemented groups was higher than in the 0% group (p < 0.05). The tibiotarsal index, Ca, and P in the tibia showed a linear effect with increased FP supplementation (p < 0.05). Moreover, the tibiotarsus weight/length index (TWLI) showed a quadratic effect with increased FP supplementation (p < 0.05). The optimal supplementation of FP for WG, V/C in the jejunum and ileum, and TWLI was 1.8, 1.9, and 1.6%, respectively. In conclusion, dietary supplementation with 1 to 2.5% LYS1 FP in broilers can improve their growth performance and the development of intestinal villi. Moreover, 1.9% is the optimal supplementation of LYS1 FP in the diet, based on the fitting results obtained with the BLQ model. [ABSTRACT FROM AUTHOR]