Bacillus Subtilis Research Articles

Generation of a synthetic autochthonous microbiota responsible for the essential flavors and brewing properties of Daqu

Based on the 26 strains of medium–high-temperature Daqu, seed microbial inocula and mature microbial inoculants were prepared and inoculated into crushed wheat to prepare cleaner medium–high-temperature Daqu (XQ). The physicochemical properties, flavor, and microbial profile of XQ were studied using traditional medium–high-temperature Daqu (CQ) or industry standards as controls. The results showed that the liquefaction ability, saccharification activity, esterification activity, and moisture content of XQ were better than those of Daqu industry standards. The results of GC–MS and E-nose analyses showed that the main flavor compounds in the two types of Daqu were esters, alcohols, acids, and aldehydes, which had similar flavor profiles. Microbial sequencing revealed that the most abundant fungi in XQ were Aspergillus subflavus, Wickerhamomyces anomalus, Rhizopus oryzae, and Candida tropicalis, whereas the most abundant bacteria were Lactobacillus curvatus, Weissella cibaria, and Bacillus subtilis. Both types of Daqu contained a high abundance of W. anomalus. In addition, the COG (Clusters of Orthologous Groups of Proteins) analyses displayed that the two types of Daqu had their dominant functions and generally had the pathways and functional characteristics of organism growth and metabolism. This study laid some experimental research bases for the preparation of cleaner Daqu and could provide insights for producing safer microbial fermented foods.

Pyrrolnitrin in Pseudomonas chlororaphis strain AFS009 metabolites reduces constitutive and DMI-induced MfCYP51 gene expression in Monilinia fructicola.

Brown rot, caused by Monilinia fructicola, is one of the most economically important diseases of peach. Demethylation inhibitor (DMI) fungicides play an important part in managing brown rot in the southeastern U.S., but over the last 20 years, reduced efficacy to DMIs has been reported in field isolates overexpressing the DMI target enzyme encoding MfCYP51 gene. Metabolites of the biocontrol agent (BCA) Pseudomonas chlororaphis strain AFS009 suppressed the MfCYP51 gene in sensitive and resistant M. fructicola isolates previousely, but it is not known what molecule was responsible. The goals of this study were to determine the presence and role of pyrrolnitrin (PRN), a common metabolite of P. chlororaphis and chemical analogue to fludioxonil with antifungal activity, in the suppression of the MfCYP51 gene and to investigate if MfCYP51 expression can also be suppressed by Bacillus subtilis. High-performance liquid chromatography (HPLC) detected pyrrolnitrin at 1.75 µg/mg in P. chlororaphis metabolites formulated as Howler EVO (Howler). PRN at 0.1 µg/ml, fludioxonil at 0.1 µg/ml, and Howler applied at a dose that contained 0.1 µg/ml PRN significantly reduced the MfCYP51 gene expression at similar levels in DMI-resistant isolates. Furthermore, MfCYP51 expression in DMI-sensitive and three DMI-resistant isolates treated with Howler (88.1 µg/ml), Theia (209.5 µg/ml), propiconazole (0.3 µg/ml), or the mixture of either Howler or Theia + propiconazole revealed that Howler significantly reduced the MfCYP51 target gene expression in two of three sensitive and all three resistant M. fructicola isolates. On the other hand, Theia showed no suppressive effect and even increased the MfCYP51 gene expression level in two of three resistant isolates. In detached fruit assays on apple with a DMI resistant isolate, only the mixture of Howler + 50 µg/ml propiconazole resulted in synergism. The results indicate that suppression of MfCYP51 target gene is BCA-dependent and can be induced by pyrrolnitrin.

Bacillus subtilis EpsA-O: A novel exopolysaccharide structure acting as an efficient adhesive in biofilms

Extracellular polysaccharides are crucial components for biofilm development. Although Bacillus subtilis is one of the most characterized Gram-positive biofilm model system, the structure-function of its exopolysaccharide, EpsA-O, remains to be elucidated. By combining chemical analysis, NMR spectroscopy, rheology, and molecular modeling, high-resolution data of EpsA-O structure from atom to supramolecular scale was obtained. The repeating unit is composed of the trisaccharide backbone [→3)-β-d-QuipNAc4NAc-(1→3)-β-d-GalpNAc-(1→3)-α-d-GlcpNAc-(1]n, and the side chain β-d-Galp(3,4-S-Pyr)-(1→6)-β-d-Galp(3,4-S-Pyr)-(1→6)-α-d-Galp-(1→ linked to C4 of GalNAc. Close agreement between the primary structure and rheological behavior allowed us to model EpsA-O macromolecular and supramolecular solution structure, which can span the intercellular space forming a gel that leads to a complex 3D biofilm network as corroborated by a mutant strain with impaired ability to produce EpsA-O. This is a comprehensive structure-function investigation of the essential biofilm adhesive exopolysaccharide that will serve as a useful guide for future studies in biofilm architecture formation.

Efficacy of Bacillus spp. Isolates for Application as Biofertilizer in Tomato (Solanum lycospersicum L.) Cultivation

Certain species of Bacillus are considered rhizobacteria that promote plant growth, inhibit the growth of plant pathogens and deleterious rhizospheric microorganisms. The aim of this study was to determine the stimulatory effect of Bacillus subtilis KBM1 and Bacillus venezensis KBM1 on the growth and disease reduction of tomato plants in Côte d'Ivoire. In this respect, three substrates consisting of soil only, sawdust only and a mixture of soil and sawdust were inoculated with the two bacteria, then left to ferment for twenty-four (24) hours. The biofertilizers obtained after fermentation were used to assess their effects on the development of potted tomatoes. Growth parameters, infection rate and mortality rate of tomato plants were measured regularly during 6 weeks of cultivation. A search for tomato pathogenic fungi was carried out to demonstrate the efficacy of Bacillus strains isolates on plant health. The results obtained showed that Bacillus subtilis KBM1 and Bacillus venezensis KBM1 had a positive impact on plant growth of tomato plants grown. Infection and mortality rates of 0% were observed for tomato plants grown on the three substrates inoculated with these two bacteria. No pathogenic fungi were isolated from the organs of tomato plants grown on the three substrates fermented with the bacteria. However, on untreated control plants, a variety of pathogenic fungi, namely Fusarium sp, Phytophtora sp, Cladosporium sp, Rhizoctonia sp, Colletotrichum sp, Penicillium sp, Rhizopus sp and Trichoderma sp were observed. However, the Bacillus subtilis KBM1 treatment stood out from Bacillus venezensis KBM1 with the best results for the growth parameters measured. In view of these results, these two bacteria could be used to produce an effective biofertilizer for tomato cultivation in Côte d'Ivoire.

Potentiality of Formulated Pseudomonas Fluorescens and Bacillus Subtilis for the Management of Sheath Blight Disease of Rice Under Field Condition

Rice production is severely affected by sheath blight disease in Bangladesh which is caused by a fungus Rhizoctonia solani that is still merely managed by synthetic chemical fungicides. In this study, different isolates of Pseudomonas fluorescens and Bacillus subtilis were evaluated to assess their inhibitory effects against R. solani both in- vitro and field condition following dual culture method in order to formulate technique for the management of sheath blight of rice. Among all the isolates, P. fluorescens (Pf- 7 and Pf- 8) and B. subtilis (Bs-17 and Bs-21) showed the highest in –vitro mycelial growth suppression of the pathogen. Based on the in-vitro mycelial growth suppression performance, fourteen treatments combinations, either single or in consortia of antagonistic bacteria in talc-based formulations were applied by root dipping. All the formulated bacterial isolates showed significant reduction in disease and severity compared to untreated control and chemical fungicide. At 90 DAT, both the disease incidence and severity were found lowest in T11 (31.67 % and 37.33 % respectively) where the seedlings were treated for 24 hours with consortium of Pf-7 + Pf- 8 formulated in talc followed by in T12 (32.00 % and 38.00 %) where the seedlings were treated for 24 hours with consortium of Bs-17 + Bs-21 formulated in talc respectively. Vegetative and yield parameters were also significantly increased in both T11 and T12. The highest yield as well as Benefit Cost Ratio (BCR) were found in T11 (2.50 kg/ m2 and 2.20) followed by T12 (2.49 kg/m2 and 2.18). Thus, the consortia of bacterial isolates (Pseudomonas fluorescens and Bacillus subtilis) with seedling dipping time for 24 hour represents a promising alternative to chemical fungicides for the management of sheath blight of rice in field condition. J Bangladesh Agril Univ 22(3): 342-351, 2024

A simple and highly efficient catalyst-free for the synthesis of dihydro pyrido[1, 2-a]quinoxaline derivatives by four component reactions and evaluation of their antibacterial activity

In the current work, a new series of methyl-10-amino-9-cyano-6-oxo-8-aryl-6,8-dihydro-5H-pyrido[1,2-a]quinoxaline-7-carboxylate derivatives were efficiently designed and prepared using a convenient route in excellent yields by four-component reactions among benzene-1,2-diamine, dimethyl acetylenedicarboxylate, various aromatic aldehydes and malononitrile. These reactions were carried out in acetonitrile at ambient temperature under catalyst-free conditions for 7 h. The structures of the new obtained compounds were confirmed by NMR, IR, EI-MS and elemental analysis. The antibacterial activity of the synthesized products was studied using bacterial strains: Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. The results showed that all the synthesized compounds are effective against Staphylococcus aureus, Bacillus subtilis and Escherichia coli bacteria.

In vitro and in vivo evaluation of the nutritional value of various hydrolyzed feather meals and comparison of their effect on performance and carcass characteristics of broilers

In the current study, the nutritional value of various hydrolyzed feather meals (HFM) was investigated in two separate experiments (an in vitro and an in vivo experiment). The in vitro experiment was based on a completely randomized design with five replications and seven treatments including (1) Raw feather meal (RFM), (2) HFM by autoclave (Au-HFM), (3) Fermented feather meal (FFM) by Bacillus licheniformis (Bl-FFM), (4) FFM by Bacillus subtilis (Bs-FFM), (5) FFM by Aspergillus niger (An-FFM), (6) FFM by Bacillus licheniformis + Bacillus subtilis + Aspergillus niger (Co-FFM), and (7) HFM by an enzyme (En-HFM). The highest in vitro pepsin-pancreatin and apparent ileal CP digestibility were observed in Co-FFM, and the lowest amount belonged to RFM (P < 0.05). For the in-vivo experiment, 480 1-d-old male Ross 308 broilers were distributed in the experimental units in a completely randomized design with 8 treatments and 5 replicates (12 chicks/replicate). The treatments were: (1) Control diet (without feather meal (FM)), (2), (3), (4), (5), (6), (7), and (8), diets containing 4 % RFM, Au-HFM, Bl-FFM, Bs-FFM, An-FFM, Co-FFM, and En-HFM, respectively. For the in-vivo study, the birds fed control and Co-FFM diets had the highest feed intake, body weight gain, and the lowest feed conversion ratio compared to the other treatments (P < 0.05). The broilers fed the control and FFMs diets had the lowest relative weight of abdominal fat and liver compared to the other groups (P < 0.05). Therefore, our findings advise the poultry feed industry to look for Co-FFM as an effective alternative and cheaper feed ingredient to replace part of soybean meal in poultry diet.

Sustainable management of blister and grey blight diseases of tea using antagonistic and plant growth promoting microbes in Western Ghats of India

The blister and grey blights are the devastating leaf diseases in tea. The severity of blister and grey blight diseases in tea ranged from 22% to 68% and 11.41%–57.02%, respectively in a survey conducted in Southern India during 2022–2023. The morphological and molecular identifications of pathogens revealed that Exobasidium vexans and Neopestalotiopsis clavispora were responsible for causing blister and grey blights on tea in southern India. The conventional management strategy with foliar spray of synthetic fungicides result in fungicidal residue in tea causes environmental pollution and health hazards to human. To overcome this the current study aims to manage the blister and grey blight diseases of tea through the microbial biocontrol agents. The bacterial biocontrol agents viz., Bacillus subtilis (BBV57), Bacillus amyloliquefaciens (TNAU), Pseudomonas fluorescens (Pf1) and the fungal antagonist, Trichoderma asperellum (TV 1) were evaluated for their efficacy against the both blight diseases. All the bio agents have significantly reduced the basidiospore germination and the mycelial growth of E. vexans and N. clavispora, respectively. Among the bio agents, B. amyloliquefaciens registered 88.36% reduction in the germination of E. vexans basidiospore and 85.56% of mycelial growth reduction of N. clavispora in vitro (P < 0.05). Under field conditions the combined soil and foliar application of bacterial antagonists viz., B. subtilis, B. amyloliquefaciens and P. fluorescens each with 1 ˟ 1011 CFU/ml @ 5% concentration at 7 days interval significantly reduced the blister blight incidence with 53.53% & 51.53% reduction over control and 51.53% & 43.65% of reduction over initial during first and second year, respectively. The combined application also reduced the grey blight incidence with 53.53% & 52.00% reduction over control and 56.81% & 47.65% of reduction over initial during first and second year, respectively. This treatments with bioagents were almost comparable with synthetic treatments, such as copper oxy chloride (0.25%) and hexaconazole (0.25%) with 64.63% and 64.76% reduction of blister and grey blight reduction in first year; 59.74%, 51.11% reduction in second year over control (P < 0.05), respectively. Thus, the application of B. subtilis, B. amyloliquefaciens and P. fluorescens can be recommended for the sustainable management of tea blister and grey blight diseases in tea gardens.

Complex sporulation-specific expression of transcription termination factor Rho highlights its involvement in Bacillus subtilis cell differentiation

Termination factor Rho, responsible for the main factor-dependent pathway of transcription termination and the major inhibitor of antisense transcription, is an emerging regulator of various physiological processes in microorganisms. In Gram-positive bacterium Bacillus subtilis, Rho is involved in the control of cell adaptation to starvation and, in particular, in the control of sporulation, a complex differentiation program leading to formation of a highly resistant dormant spore. While the initiation of sporulation requires a decrease in Rho protein levels during the transition to stationary phase, the mechanisms regulating the expression of rho gene throughout the cell cycle remain largely unknown. Here we show that a drop in the activity of the vegetative SigA-dependent rho promoter causes the inhibition of rho expression in stationary phase. However, after the initiation of sporulation, rho gene is specifically reactivated in two compartments of the sporulating cell using distinct mechanisms. In the mother cell, rho expression occurs by read-through transcription initiated at the SigH-dependent promoter of the distal spo0F gene. In the forespore, rho gene is transcribed from the intrinsic promoter recognized by the alternative sigma factor SigF. These regulatory elements ensure the activity of Rho during sporulation, which appears important for the proper formation of spores. We provide experimental evidence that disruption of the spatiotemporal expression of rho during sporulation affects the resistance properties of spores, their morphology, and the ability to return to vegetative growth under favorable growth conditions.

Investigating the in vitro antibacterial, antibiofilm, antioxidant, anticancer and antiviral activities of zinc oxide nanoparticles biofabricated from Cassia javanica.

It is thought to be risk-free, environmentally benign, and safe for biological processes to produce zinc oxide nanoparticles from renewable resources. This study examined Cassia javanica's ability to create ZnONPs. The generated ZnONPs were analyzed using a variety of techniques, such as TEM, FTIR spectroscopy, UV-Vis spectroscopy, and XRD analysis. The antibacterial potential of ZnONPs has been investigated using both Agar well diffusion and microtitreplate (MTP) methods. One method used to evaluate ZnONPs' capacity to scavenge free radicals at different concentrations was the DPPH method. The permanent zinc oxide (ZnO) shape and the naturally occurring crystal structure of ZnONPs were validated by the XRD data. ZnONPs showed antibacterial activity with MICs of 31.7 μg/mL toward Bacillus subtilis, 62.5 μg/mL for Salmonella typhimurium, Escherichia coli while Clostridium sporogenes and Bacillus pumilus was 125μg/mL. Furthermore, ZnONPs demonstrated a range of antibiofilm activities toward Staphylococcus aureus (MRSA). ZnONPs showed an intriguing antioxidant capacity, achieving IC50 of 109.3 μg/ml μg/mL. Additionally, ZnONPs demonstrated low toxic effect on Vero cell with IC50 154.01 μg/mL as well as possible anticancer action when applied to the carcinoma cell lines HepG2 with IC50 of 47.48 μg/mL. Furthermore, ZnONPs at 62.5 μg/mL had a promising antiviral impact against HSV1 and COX B4, with antiviral activities of 75.4% and 65.8%, respectively.

Antioxidant, antibacterial, and α-glucosidase inhibition potential of three Allium species (Amaryllidaceae) from Iran

Introduction: Wild species of the genus Allium have high potential for use as medicine due to their essential secondary metabolites with antioxidant activity. This study explored the antioxidant, antibacterial, and α-glucosidase inhibition activities of three Allium species: Allium tripedale, Allium hooshidaryae, and Allium stipitatum. Methods: The antioxidant potentials of the plant methanol extracts were evaluated using the ferric reducing antioxidant power (FRAP) and the 2,2-diphenyl-1picrylhydrazil (DPPH) radical scavenging test. Total phenolic content (TPC), total flavonoid content (TFC) and α-glucosidase inhibition were also evaluated. Antibacterial assessments were done employing disk diffusion and microdilution methods to determine inhibition zone and minimum inhibitory concentration (MIC), respectively against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. Results: Allium hooshidaryae displayed high TPC (70.24 ± 0.0039 mg gallic acid equivalent/g extract), while A. tripedale had the highest TFC (87 ± 0.013 mg Quercetin equivalent/g extract). A. hooshidaryae showed superior antioxidant capacity (DPPH IC50: 724.4 ± 0.31 µg/mL; FRAP: 36.87 mg ascorbic acid equivalent/g extract) and stronger α-glucosidase inhibition (IC50 = 2.59 mg/mL vs. 4.33 mg/mL for A. tripedale and 6.41 mg/mL for A. stipitatum). Qualitative tests confirmed phenolic, flavonoid, and glycoside compounds in all three species. A. stipitatum uniquely contained saponin and tannin. A. hooshidaryae and A. stipitatum inhibited the bacterial strains effectively, especially at the higher concentration (400 µg/mL). A. stipitatum showed inhibition against all strains, particularly against S. aureus (MIC: 12.5 µg/mL). Conclusion: This study highlights the antidiabetic and antibacterial potential of three Allium species, emphasizing their values as rich sources of bioactive compounds.

Exploring the uncharted: Zinc and phosphate solubilization in Zn-P isolates from wheat rhizosphere inceptisols

This research delves into the untapped potential of phosphorous and zinc solubilizing rhizobacteria, known as Zn-P isolates, from wheat rhizosphere inceptisols. A total of thirty rhizosphere soil samples were collected, resulting in recovery of forty unique bacterial isolates. After initial screening, out of forty isolates, recovered on the basis of halozone formation on the nutrient agar medium. four were subjected to biochemical and further molecular identification. Four isolates, identified as Bacillus subtilis (Zn-P-1), Pseudomonas aureginosa (Zn-P-2), Staphylococcus aureus (Zn-P-3), and Methylobacterium organophyllum (Zn-P-4) through a set of 16S rRNA primers, forward (5́- GGATGAGCCCGCGGCCTA-3́) and reverse (5́- CGGTGTGTACAAGGCCCGG-3́), exhibited superior solubilization efficiency of phosphorous and zinc These strains were evaluated via in vitro and pot culture assays. The study found that Zn-P-1 demonstrated the highest zinc solubilization (134.87 mg/l) when ZnO was used as the zinc source as compared to ZnCO3 and Zn-EDTA as also highest at 72.32 mg/l in CaH2PO4 and lowest at 14.44 mg/l with KH2PO4 using P sources, thus highlighting the role of Bacillus in zinc and phosphorous activity with substrate type. The inoculation of tri-calcium phosphate (TCP) and ZnCO3, along with Bacillus and Methylobacterium, led to increased phosphorous and zinc solubilization, uptake, and use efficiency, marking these rhizobacteria as potentially beneficial for nutrient enhancement and PGPR activities in wheat crops grown in inceptisols.

Biological Control of Anthracnose in Capsicum annuum Using Inoculants in Hydroponics: A Mechanistic Review

The Colletotrichum species that causes anthracnose poses a significant threat to the growth of Capsicum annuum, or ringer pepper, resulting in significant yield losses. Conventional produced control systems can give rise to ecological and economic difficulties, necessitating the need for alternative, workable solutions. This study explores the feasibility and parts of natural inoculants in controlling anthracnose ailment in Capsicum annuum filled in chipped away at hydroponics structures. As organic inoculants, we utilized a wide variety of useful microorganisms, such as Trichoderma harzianum, Bacillus subtilis, and mycorrhizal parasites. At various stages of the Capsicum annuum plant's development, these inoculants were incorporated into the supplement arrangements for tank farming. Anthracnose was observed and compared to untreated controls and synthetically treated plants in terms of its severity. Plants treated with natural inoculants show a significant reduction in sickness rate and severity, as shown by our findings. In fact, Trichoderma harzianum outperformed Bacillus subtilis and mycorrhizal parasites in terms of biocontrol productivity. Through a series of biochemical and sub-atomic tests, the fundamental instruments were examined. Natural inoculants, according to important discoveries, increase the plant's intrinsic safe reaction by causing fundamental obstruction. This is affirmed by the upregulation of pathogenesis-related (PR) characteristics and extended activity of shield impetuses like peroxidase (Unit), polyphenol oxidase (PPO), and phenylalanine antacid lyase (Amigo). In addition, the inoculants improved root development and supplement absorption, enhancing overall establish health and resistance to microorganisms. This study demonstrates that natural inoculants can be a reasonable and effective method for controlling anthracnose in Capsicum annuum grown in improved tank farms. In accordance with the goals of a potential agribusiness, incorporating these biocontrol specialists into aquafarming frameworks can increase crop productivity and reduce the use of compound pesticides.

Lag Plot: A Novel Method to Determine Viable Bacterial Cell Number via a Single OD Measurement.

Bactericidal activity is a valuable parameter which is considered and measured for antimicrobial compounds. The available standard protocol to evaluate bactericidal activity is based on the direct colony count. Colony counting requires serial dilution, plating, overnight incubation, and direct counting, which is time-and labor-intensive. In regard to eliminate direct plate count, novel techniques were developed based on the real-time growth monitoring which can come with some limitations and drawbacks. These drawbacks encourage us to develop a novel technique with simple procedure to determine viable bacterial cell count. In this procedure, real-time growth monitoring is not required. In fact, after an incubation time, the number of viable bacteria can be determined with a single OD measurement and through an equation. In this regard, four standard bacterial strains, including Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), and Bacillus subtilis (ATCC 23857), were cultured with descending inoculum densities (1.5 × 107 to 15CFU/mL) and growth curves were drawn. As expected, growth in the samples with lower inoculum densities recorded with longer lag phase. Also, a direct relation was observed between the recorded turbidities and initial cell counts. A logarithmic curve (lag plot) was obtained by plotting the OD, after 12-18h incubation, against initial cell counts. In all examined strains, R2 was calculated in the range of 0.96-0.99 which is acceptable value for coefficient of determination. Equation corresponding to the lag plot was obtained and called lag equation. This equation is applicable to calculate the number of viable cells in unknown samples simply by inoculation, incubation, and single OD measurement. Developed technique can be introduced as a simple substitution for labor- and time-intensive direct colony counting.

Reactivity of (3‐(4‐Fluorophenyl)oxyran‐2‐yl)(naphthalene‐2‐yl)methanone Toward Some Nucleophiles for Preparing Promising Agents with Antimicrobial and Antioxidant Activities

Abstract(3‐(4‐Fluorophenyl)oxyran‐2‐yl)(naphthalene‐2‐yl)methanone 2 was prepared from the reaction of 3‐(4‐fluorophenyl)‐1‐(naphthalen‐2‐yl)prop‐2‐en‐1‐one (1) with hydrogen peroxide and reacted with different nucleophiles such as hydrazine hydrate, phenyl hydrazine, thiosemicarbazide to give 4‐hydroxy‐1H‐pyrazole derivatives 3a,b‐4. Also, compound 2 was treated with hydroxylamine hydrochloride to give the substituted 2,3‐dihydroisoxazole 5. Similarly, compound 2 was treated with carbon disulfide, phenyl isothiocyanate, and phenyl isocyanate to give the corresponding compounds 6–8. Various spectroscopic techniques and elemental analysis were used to determine the molecular structures of the new derivatives. All the new analogs were screened against five human pathogenic microbial strains: Escherichia coli and Pseudomonas aeruginosa as Gram‐negative bacteria, Bacillus subtilis and Staphylococcus aureus as Gram‐positive bacteria and the yeast Candida albicans using agar diffusion method. Compound 4 exhibited moderate antimicrobial activity against all the tested microorganisms compared to positive control ciprofloxacin and fluconazole. On the other hand, the free radical scavenging activity of the new compounds using DPPH assay protocol revealed that most of the compounds have a weak reducing activity. Moreover, compounds 4 and 8 were docked against 1KZN protein (DNA gyrase), 3FYV and 5V5Z showing the ability to interact with key amino acid residues and the potential to act as inhibitors.

Plants-Extracts mediated CuS Nanoparticles: An Effective Antibacterial Agent

Antibacterial activity of plant-mediated synthesized CuS nanoparticles against gram-positive Bacillus subtilis and gram negative Pseudomonas putida is studied in this presented work. A facile hydrothermal approach was followed to synthesize spherical shaped CuS nanoparticles using flowers extract of Tagetes patula (S1), leaves extract of Azadirachta indica (S2), and bark extract of Terminalia arjuna (S3). Crystal phase identification, average crystallites size, surface morphology, absorbance spectra and energy band gap analyses of samples were further determined using XRD, SEM, and UV-DRS spectroscopy respectively. Furthermore, under irradiation of visible light, antibacterial activity of marigold flowers mediated (S1), neem leaves mediated (S2), and arjuna bark mediated (S3) CuS samples were studied. XRD data confirmed the marigold mediated CuS sample has relatively smaller crystallites size of 10.67 nm. The estimated band gap energies for respective S1, S2, and S3 samples are 174 eV, 175 eV, and 177 eV. The antibacterial analysis of S1 sample displays its excellent antibacterial activity with the formation of relatively large inhibition zones of 29 mm and 30 mm diameters, against Bacillus subtilis and Pseudomonas putida respectively. The demonstrated antibacterial activity of biocompatible CuS nanoparticles suggests their potential use in biomedical applications.

Investigation of Acetoin Biosynthesis by Bacillus subtilis ACA-DC 1176 Growing on Crude Glycerol in Flask and Bioreactor Trials

Acetoin biosynthesis by two Bacillus subtilis strains valorising crude glycerol was thoroughly explored within a pre-defined range of culture conditions and systems. B. subtilis ACA-DC 1176 stood out for its higher efficiency in acetoin production, prompting an investigation into the potential for enhanced productivity through the evaluation of diverse culture conditions and media compositions. The primary by-products of the biodiesel and corn industries, namely crude glycerol and corn steep liquor, respectively, were successfully employed as the principal carbon and nitrogen sources of the newly developed low-cost culture medium. Furthermore, the results of the various feeding strategies that were tested indicated that the conversion of 2,3-butanediol (BDO) to acetoin occurred exclusively when the concentration of glycerol was below approximately 5 g/L. This seemed to be necessary for the production of NADH, which is essential for maintaining cellular processes. Following the complete depletion of glycerol, acetic acid increased and became the predominant metabolite, while both acetoin and BDO decreased, presumably resulting in ATP generation. This is likely a mechanism employed by the cell to generate energy in the absence of a carbon source. In the fed-batch bioreactor culture, the kinetics of metabolites differed, as there was no conversion of BDO to acetoin at the final depletion of glycerol. At volumetric mass transfer coefficient (kLa) levels exceeding approximately 70 1/h, the production of acetoin was favoured over that of BDO, with the highest observed acetoin/BDO ratio reaching 4.29 g/g. Conversely, at kLa values below approximately 60 1/h, the titres of acetoin and BDO were found to be nearly equal. The final concentrations of acetoin and BDO reached 36.0 g/L and 25.5 g/L, respectively, resulting in a total yield of both (acetoin + BDO) per glycerol consumption of 0.40 g/g. To the best of our knowledge, this is the first study to focus on acetoin production from crude glycerol fermentative valorisation. The study presents new findings regarding the parameters influencing the level of BDO conversion to acetoin. However, further research is required in order to gain a comprehensive understanding of the underlying phenomena and metabolic pathways involved.

Clarithromycin-tailored cubosome: A sustained release oral nano platform for evaluating antibacterial, anti-biofilm, anti-inflammatory, anti-liver cancer, biocompatibility, ex-vivo and in-vivo studies

The clinical implication of clarithromycin (CLT) is compromised owing to its poor solubility and, subsequently, bioavailability, unpalatable taste, rapid metabolism, short half-life, frequent dosing, and adverse effects. The present investigation provides an innovative sustained-release oral drug delivery strategy that tackles these challenges. Accordingly, CLT was loaded into a cubosome, a vesicular system with a bicontinuous cubic structure that promotes solubility and bioavailability, provides a sustained release system combating short half-life and adverse effects, masks unpleasant taste, and protects the drug from destruction in gastrointestinal tract (GIT). Nine various formulas were fabricated using the emulsification method. The resulting vesicles increased the encapsulation efficiency (EE %) from 57.64 ± 0.04 % to 96.80 ± 1.50 %, the particle size (PS) from 147.30 ± 21.77 nm to 216.61 ± 5.37 nm, and the polydispersity index (PDI) values ranged from 0.117 ± 0.024 to 0.278 ± 0.073. The zeta potential (ZP) changed from −20.65 ± 2.01 mV to −33.98 ± 2.60 mV. Further, the release profile exhibited a dual release pattern within 24 h., with the percentage of cumulative release (CR %) expanding from 30.06 ± 0.42 % to 98.49 ± 2.88 %, optimized formula was found to be CC9 with EE % = 96.80 ± 1.50 %, PS = 216.61 ± 5.37 nm, ZP = −33.98 ± 2.60 mV, PDI = 0.117 ± 0.024, CR % = 98.49 ± 2.88 % and IC50 of 0.74 ± 0.19 µg/mL against HepG-2 cells with scattered unilamellar cubic non-agglomerated vesicles. Additionally, it exhibited higher anti-MRSA biofilm, relative bioavailability (2.8 fold), and anti-inflammatory and antimicrobial capacity against Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus compared to free CLT. Our data demonstrate that cubosome is a powerful nanocarrier for oral delivery of CLT, boosting its biological impacts and pharmacokinetic profile.

Isolation of a novel Bacillus subtilis HF1 strain that is rich in lipopeptide homologs and has strong effects on the resistance of plant fungi and growth improvement of broilers.

Bacillus subtilis is an important probiotic microorganism that secretes a variety of antimicrobial compounds, including lipopeptides, which are a class of small molecule peptides with important application value in the fields of feed additives, food, biopesticides, biofertilizers, medicine and the biological control of plant diseases. In this study, we isolated a novel B. subtilis HF1 strain that is rich in lipopeptide components and homologs, has a strong antagonistic effect on a variety of plant fungi, and is highly efficient in promoting the growth of broilers. The live B. subtilis HF1 and its fermentation broth without cells showed significant inhibitory effects on 20 species of plant fungi. The crude extracts of lipopeptides in the fermentation supernatant of B. subtilis HF1 were obtained by combining acid precipitation and methanol extraction, and the lipopeptide compositions were analyzed by ultrahigh-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The results showed that HF1 could produce 11 homologs of surfactin and 13 homologs of fengycin. Among the fengycin homologs, C13-C19 fengycin A and C15-C17 fengycin B were identified; among the surfactin homologs, C11-C17 surfactin A and C13-C16 surfactin B were characterized. C13 fengycin A, C11 surfactin A and C17 surfactin A were reported for the first time, and their functions are worthy of further study. In addition, we found that HF1 fermentation broth with and without live cells could be used as a feed additive to promote the growth of broilers by significantly increasing body weight up to 15.84%. HF1 could be a prospective strain for developing a biocontrol agent for plant fungal diseases and an efficient feed additive for green agriculture.

Enhanced phosphorus availability and cadmium remediation using phosphate-solubilizing bacteria-loaded biochar in contaminated soils

Phosphate-solubilizing bacteria (PSB) have garnered extensive attention for their ability to improve soil phosphorus (P) availability and their potential applications in bioremediation. However, the efficacy of PSB is contingent upon their colonization in soils, which can be hindered by the toxicity of heavy metals such as cadmium (Cd). Utilizing biochar as a protective carrier for PSB presents a promising strategy to address this challenge. This study investigated the effects of PSB-loaded biochar on soil P pools and wheat (Triticum aestivum L.) growth under Cd-contaminated conditions. Results indicated that biochar loading facilitated the colonization and functioning of the inoculated PSB strain (Bacillus subtilis), as evidenced by increased soil cultivatable PSB abundance and phosphatase activity as well as increased relative abundance of Bacillus in bacterial community. Consequently, improved soil P availability and reduced soil exchangeable Cd content were observed, leading to a 30.3% increase in wheat shoot biomass, a 50.4% increase in P accumulation, and a 24.1% reduction in Cd accumulation in wheat shoots under the PSB-loaded biochar treatment. Significant shifts in bacterial community composition were revealed by 16S rRNA amplicon sequencing and enhanced microbe-mediated metabolic activities were observed, as demonstrated by soil enzyme assays and metabolome analysis. Overall, these findings underscore the potential of PSB-loaded biochar to enhance P availability, mitigate Cd toxicity, and shape the soil microbial community, offering a promising strategy for sustainable nutrient management and bioremediation in Cd-contaminated agricultural soils.