Bacillus Subtilis Strain Research Articles

Evaluation of antimicrobial activity and bioactive compound analysis of Verbascum thapsus L. A folklore medicinal plant

BackgroundThe prevalence of microbial infectious diseases is a major contributor to morbidity and mortality on a global scale due to the emergence of antibiotic-resistant microbes, despite the fact that many new antibiotics have been introduced for several decades. In present scenario, the potential utilization of natural plant derived extracts for medicinal and therapeutic purposes has increased remarkably due to adverse effects of synthetic drugs and resistance among pathogenic microbes. Aim of the studyVerbascum thapsus L. (Common mullein) has been traditionally used as a medicine for lung, skin and throat and many other disorders since time immemorial, therefore, the current study was aimed to evaluate the antimicrobial activity of leaf and root extracts of Verbascum thapsus against pathogenic bacterial and fungal strains and its phytochemical composition by spectral studies. MethodsSoxhlet extraction method was employed for extraction process by using varying polarity solvents like hexane, ethyl acetate and methanol. Phytochemical analysis of methanol extract was determined by Fourier Transform-Infrared Spectroscopy (FTIR) and Gas Chromatography-Mass Spectrometry (GC–MS). Antimicrobial activity was evaluated by determining the MIC and MBC/MFC of each extract by broth dilution method. ResultsThe antimicrobial competence of each extract sample was evaluated against six pathogenic bacterial strains and three fungal strains. Methanolic extract of leaves and the ethyl acetate extract of roots of Verbascum thapsus exhibited significant zones of inhibition against all the tested microorganisms. The highest inhibition zone was observed against the bacterial strain Bacillus subtilis (24.1±0.18 mm) followed by Streptococcus pyogenes (22.9±0.64 mm) and the fungal strain Aspergillus niger (22.8±0.55 mm) at a concentration of 500 μg/ml. Methanolic leaf extract strongly inhibited the growth of tested microbial strains with MIC ranged from 15.625 to 125 μg/ml. The FTIR and GCMS analysis confirms the presence of bioactive compound, possessing broad range of therapeutic activities. ConclusionThis Presence of various important phytoconstituents and broad-spectrum antimicrobial activity points Verbascum thapsus as a promising source for the isolations of molecules responsible for antimicrobial activities.

Probiotic Potential of Bacillus Subtilis Strain I3: Antagonistic Activity Against Chalkbrood Pathogen and Pesticide Degradation for Enhancing Honeybee Health.

To explore the potential of probiotic candidates beneficial for honeybee health through the modulation of the gut microbiome, bee gut microbes were isolated from bumblebee (Bombus terrestris) and honeybee (Apis mellifera) using diverse media and cultural conditions. A total of 77 bee gut bacteria, classified under the phyla Proteobacteria, Firmicutes, and Actinobacteria, were identified. The antagonistic activity of the isolates against Ascosphaera apis, a fungal pathogen responsible for chalkbrood disease in honeybee larvae, was investigated. The highest growth inhibition percentage against A. apis was demonstrated by Bacillus subtilis strain I3 among the bacterial strains. The presence of antimicrobial peptide genes in the I3 strain was detected using PCR amplification of gene fragments encoding surfactin and fengycin utilizing specific primers. The export of antimicrobial peptides by the I3 strain into growth medium was verified using liquid chromatography coupled with mass spectroscopy. Furthermore, the strain's capabilities for degrading pesticides, used for controlling varroa mites, and its spent growth medium antioxidant activity were substantiated. The survival rate of honeybees infected with (A) apis was investigated after feeding larvae with only medium (fructose + glucose + yeast extract + royal jelly), (B) subtilis I3 strain, A. apis with medium and I3 strain + A. apis with medium. Honeybees receiving the I3 strain + A. apis exhibited a 50% reduction in mortality rate due to I3 strain supplementation under experimental conditions, compared to the control group. In silico molecular docking revealed that fengycin hydrolase from I3 strain effectively interacted with tau-fluvalinate, suggesting its potential in bee health and environmental protection. Further studies are needed to confirm the effects of the I3 strain in different populations of honey bees across several regions to account for genetic and environmental variations.

The Biological Efficiencies of Multifunctional Complexes Based on Bacillus subtilis Strains and Chitosan Salicylate in Wheat Cultivation

The Biological Efficiencies of Multifunctional Complexes Based on Bacillus subtilis Strains and Chitosan Salicylate in Wheat Cultivation

Production of ultra-high-molecular-weight poly-γ-glutamic acid by a newly isolated Bacillus subtilis strain and genomic and transcriptomic analyses.

Poly-γ-glutamic acid (γ-PGA) is a microbial-derived polymer with molecular weight (Mw) from 104 to 107 Da, and the high-Mw(>7.0 × 105 Da) or ultra-high-Mw(>5.0 × 106 Da) γ-PGA has important application value as a tissue engineering material, as a flocculant, and as a heavy metal remover. Therefore, how to produce these high-Mw γ-PGAs with low cost and high efficiency has attracted wide attention. In this study, a γ-PGA producer was isolated from the natural environment, and identified and named Bacillus subtilis GXD-20. Then, the ultra-high-Mw(>6.0 × 106 Da) γ-PGA produced by GXD-20 was characterized. Interestingly, GXD-20 could produce γ-PGA at 42°C, and exhibited a γ-PGA titer of up to 22.29±0.59 g L-1 in a 5-L fermenter after optimization of the fermentation process. Comparative genomic analysis indicated that the specific protein sequence and subcellular localization of PgdS (a γ-PGA-degrading enzyme) were closely related to the ultra-high-Mw of γ-PGA. Transcriptomic analysis revealed that the high γ-PGA titer at 42°C was mainly related to the high expression of genes encoding enzymes for sucrose transportation and utilization, nitrogen transportation, endogenous glutamate synthesis, and γ-PGA synthesis. These results provide new insights into the production of ultra-high-Mw γ-PGA by Bacillus at high temperatures.

Using kin discrimination to construct synthetic microbial communities of Bacillus subtilis strains impacts the growth of black soldier fly larvae.

Using synthetic microbial communities to promote host growth is an effective approach. However, the construction of such communities lacks theoretical guidance. Kin discrimination is an effective means by which strains can recognize themselves from non-self, and construct competitive microbial communities to produce more secondary metabolites. However, the construction of cooperative communities benefits from the widespread use of beneficial microorganisms. We used kin discrimination to construct synthetic communities (SCs) comprising 13 Bacillus subtilis strains from the surface and gut of black soldier fly (BSF) larvae. We assessed larval growth promotion in a pigeon manure system and found that the synthetic community comprising 4 strains (SC 4) had the most profound effect. Genomic analyses of these 4 strains revealed that their complementary functional genes underpinned the robust functionality of the cooperative synthetic community, highlighting the importance of strain diversity. After analyzing the bacterial composition of BSF larvae and the pigeon manure substrate, we observed that SC 4 altered the bacterial abundance in both the larval gut and pigeon manure. This also influenced microbial metabolic functions and co-occurrence network complexity. Kin discrimination facilitates the rapid construction of synthetic communities. The positive effects of SC 4 on larval weight gain resulted from the functional redundancy and complementarity among the strains. Furthermore, SC 4 may enhance larval growth by inducing shifts in the bacterial composition of the larval gut and pigeon manure. This elucidated how the SC promoted larval growth by regulating bacterial composition and provided theoretical guidance for the construction of SCs.

A molybdenum-ciprofloxacin complex with an isopolyoxometalate having DNA binding and antibacterial activity

In this study, an antibacterial complex based on the quinolone antibacterial drug ciprofloxacin (CF) and a polyoxometalate was synthesized, Mo2O5(CF)2(CFd)2(β-Mo8O26)·5H2O (1). Complex 1 was characterized by elemental analysis and X-ray single-crystal diffraction. Single-crystal structural analysis revealed that 1 was made of β-Mo8 clusters, binuclear molybdenum clusters Mo2O5(CF)2(CFd)2, and water molecules. The antibacterial activity of 1 has been tested against three microorganisms (Staphylococcus aureus, Escherichia coli, and Bacillus subtilis strains); the antibacterial activities of 1 were higher than those of free CF at the same mass amounts. The interaction between 1 and calf thymus DNA (CT-DNA) was investigated using Uv-vis spectroscopy to determine the binding constants. The results indicated that intercalation is the probable mode of action, with 1 demonstrating higher binding constants (K b) to CT-DNA (1.8 × 105 M−1) compared to pure CF (4.9 × 104 M−1). These findings show that the implementation of molybdate for ciprofloxacin modification resulted in the formation of a metal complex with enhanced antibacterial activity over CF and significantly improving its binding affinity to DNA.

Microbial inoculation alters rhizoplane bacterial community and correlates with increased rice yield

Microbial inoculants are recognized as environmentally friendly methods to promote plant growth and improve soil properties. However, the effects of inoculation on the rhizosphere and rhizoplane community structure of plants remain poorly documented and need further investigation. Rhodopseudomonas palustris (R. palustris) strain has nitrogen fixing ability and Bacillus subtilis (B. subtilis) strain is a plant growth-promoting rhizobacterium (PGPR). In this study, we investigated the effects of single and co-inoculation with R. palustris and B. subtilis on the increase of rice yield as well as on the microbial communities in the rhizosphere and rhizoplane of rice through potting experiments, respectively. The results showed that inoculation significantly increased rice yield and seed setting rate, with co-inoculation raising the yield by up to 13.7%. Inoculation influenced both rhizosphere and rhizoplane community structures and functions, amplifying the differences between them. The most significant changes were brought about by the combined inoculation treatment. Co-inoculation with R. palustris and B. subtilis had a synergistic effect. The profound alterations of rhizoplane bacterial community structures and functions were proved to be positively correlated with rice yield and seed setting rate (r = 0.59–0.76, p < 0.05). These results provide new ideas for the investigation of the potential microbiological mechanisms of microbial co-inoculation in practical agricultural applications.

Biodegradation of sugarcane trash into substrates of utilizable products using lignocellulolytic microorganisms

Biodegradation of sugarcane trash was conducted using lignocellulolytic microorganisms, Bacillus subtilis (MW715021) and Schizophyllum commune (MW720154) strains. Throughout the biodegradation process, various physicochemical, biochemical, and microbial parameters were monitored. The weight, organic carbon, and C:N ratio of the sugarcane residue progressively decreased from 20 to 120 days of incubation. Among different treatments, T7 (Sugarcane trash + Bacillus subtilis + Schizophyllum commune) exhibited significantly the lowest weight, organic carbon, and C:N ratio of 61.60 g, 17.85%, and 13.02, respectively. The pH of the substrate decreased slightly at the initial period of biodegradation then it increased slightly and became stable at near about neutral pH. The Electrical Conductivity (EC) of the substrate was increased with the advancement of incubation periods. The Total Kjeldahl Nitrogen (TKN) content of the substrate increased with the duration of incubation. A higher TKN content of 1.37% was recorded in T7 on 120 days of incubation. The nutrient content (P2O5, K2O, Fe, Mn, Zn, and Cu) of the substrate was increased with the advancement of the decomposition period. The cellulose and lignin content was reduced from 40.78 to 21.54 % to 23.16 and 10.84 %, respectively in T7. Lignocellulolytic enzymes i.e. cellulase, laccase, lignin peroxidase, and manganese peroxidase were increased at initial degradation periods up to 60 days then it declined at a later stage of biodegradation. The microbial population increased with incubation periods up to 60 days then after it was declined at a later stage of biodegradation.

Optimization of the Conditions for the Transformation of a Bacillus subtilis Strain L11 to Prepare Nano Selenium and Its Preliminary Application in Sheep Feed

Selenium nanoparticles (SeNPs) have greater bioavailability and safety than inorganic selenium, and was widely used in medical, agricultural, nutritional supplements, and antibacterial fields. The present study screened a strain L11 producing SeNPs from a selenium rich dairy cow breeding base in Hubei Province, China. The strain was identified as Bacillus subtilis through physiological, biochemical, and molecular biology analysis. By adjusting the cultivation conditions, the experiment determined the ideal parameters for L11 to efficiently produce SeNPs. These parameters include a pH value of 6, a cultivation temperature of 37 °C, a concentration of 4 mmol/L Na2SeO3, and a cultivation of 48 h. X-ray Photoelectron Spectroscopy (XPS), Scanning Electron Microscope-Energy Dispersive Spectroscopy (SEM-EDS), and Transmission Electron Microscopy (TEM) were used to verify that the Se particles produced by L11 are SeNPs with diameters ranging from 50 to 200 nm. The combination of the protein analysis of different cell components and TEM analysis showed that L11 mainly produces SeNPs through the transformation of the cell’s periplasmic space, cell membrane, and cell wall. Adding the L11 SeNPs complex to sheep feed can significantly enhance the antioxidant activity and immunity of sheep, and increase the Se content in the neck muscles, liver, and spleen tissues.

Effect of a Bacillus subtilis plus Yeast Cell Wall Synbiotic on Salmonella Enteritidis Colonization in Ceca of Layer Pullets

Salmonella Enteritidis (SE) is a major contamination concern in eggs and risk for Salmonellosis in humans. Strains of Bacillus subtilis and yeast cell wall can be used as substitutes for antibiotic substances in feed against Salmonella in poultry. The objective of this study was to assess the effect of BacPack® (Quality Technology International, Inc., Elgin, IL, USA) Q1+1 (BPQ11), a feed additive combination of a Bacillus subtilis strain and Saccharomyces cerevisiae cell wall, on SE cecal colonization in Lohmann LSL pullets. A control group (CON) and a test group (BPQ) were each randomly assigned 100-day-old chicks. CON was fed a corn–soybean meal-based vegetarian mash diet, and BPQ was fed the control diet supplemented with BPQ11 for the duration of the study. At 8 days of age, chicks were orally challenged with a nalidixic acid-resistant SE strain at a dose of 6.3 × 107 colony forming units (CFUs) per bird. At 7, 11, 15, and 19 days post-challenge (DPC), 25 birds per group were euthanized, and their cecal contents were collected and analyzed for SE. SE counts were 6.88, 7.98, 7.79, and 7.50 in CON and 7.18, 7.31, 6.35, and 6.30 log10 CFU/g in BPQ at 7, 11, 15, and 19 DPC, respectively. SE did not differ between CON and BPQ at 7 DPC; however, BPQ had lower (p &lt; 0.0001) SE at 11 (−0.67), 15 (−1.45), and 19 (−1.20 log10 CFU/g) DPC. Results indicate that synbiotic BPQ11 may be a useful dietary pre-harvest tool for SE management in layer birds.

XPORT ENTRAP: A droplet microfluidic platform for enhanced DNA transfer between microbial species

A significant hurdle for the widespread implementation and use of synthetic biology is the challenge of highly efficient introduction of DNA into microorganisms. This is especially a barrier for the utilization of non-model organisms and/or novel chassis species for a variety of applications, ranging from molecular biology to biotechnology and biomanufacturing applications. Common approaches to episomal and chromosomal gene editing, which employ techniques such as chemical competence and electroporation, are typically only amenable to a small subset of microbial species while leaving the vast majority of microorganisms in nature genetically inaccessible. To address this challenge, we have employed the previously described B. subtilis broad-host conjugation strain, XPORT, which was modularly designed for loading DNA cargo and conjugating such DNA into recalcitrant microbes. In this current work, we have leveraged and adapted the XPORT strain for use in a droplet microfluidic platform to enable increased efficiency of conjugation-based DNA transfer. The system named DNA ENTRAP (DNA ENhanced TRAnsfer Platform) utilizes cell-encapsulated water-in-oil emulsion droplets as pico-liter-volume bioreactors that allows controlled contacts between the donor and receiver cells within the emulsion bioreactor. This allowed enhanced XPORT-mediated genetic transfer over the current benchtop XPORT process, demonstrated using two different Bacillus subtilis strains (donor and receiver), as well as increased throughput (e.g., number of successfully conjugated cells) due to the automated assay steps inherent to microfluidic lab-on-a-chip systems. DNA ENTRAP paves the way for a streamlined automation of culturing and XPORT-mediated genetic transfer processes as well as future high-throughput cell engineering and screening applications.

Antagonistic potential of forestry compost bacteria on Sclerotinia sclerotiorum (Lib.) de Bary, causal agent of carrot white mould

Composts are known to contain beneficial bacteria, which may be antagonistic to plant pathogens. This work evaluated whether carrot white mould, causal agent Sclerotinia sclerotiorum, can be reduced using antagonistic bacteria isolated from forestry compost. In vitro and in vivo experiments demonstrated that bacteria from the genera Pseudomonas and Bacillus can inhibit mycelial growth and reduce white mould. Bacillus subtilis strains F9–2 and F9–12 and Pseudomonas arsenicoxydans strain F9–7 showed the highest inhibitory properties. Three cyclic dipeptides (diketopiperazines) were characterized from the antifungal culture filtrates of P. arsenicoxydans F9–7. When assayed against S. sclerotiorum, the diketopiperazines showed the following inhibitory activity, in increasing order: cyclo-(l-Pro-l-Val), cyclo-(l-Pro-l-Phe) and cyclo-(l-Pro-l-Leu). The combination of these diketopiperazines indicated additive and, occasionally, synergistic antifungal effects. These results indicated a potential for some bacteria to inhibit the growth of S. sclerotiorum and reduce its associated disease on carrots postharvest.

Antimicrobial Activity of Citrate-Coated Cerium Oxide Nanoparticles.

The purpose of this study was to investigate the antimicrobial activity of citrate-stabilized sols of cerium oxide nanoparticles at different concentrations via different microbiological methods and to compare the effect with the peroxidase activity of nanoceria for the subsequent development of a regeneration-stimulating medical and/or veterinary wound-healing product providing new types of antimicrobial action. The object of this study was cerium oxide nanoparticles synthesized from aqueous solutions of cerium (III) nitrate hexahydrate and citric acid (the size of the nanoparticles was 3-5 nm, and their aggregates were 60-130 nm). Nanoceria oxide sols with a wide range of concentrations (10-1-10-6 M) as well as powder (the dry substance) were used. Both bacterial and fungal strains (Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris, Candida albicans, Aspergillus brasielensis) were used for the microbiological studies. The antimicrobial activity of nanoceria was investigated across a wide range of concentrations using three methods sequentially; the antimicrobial activity was studied by examining diffusion into agar, the serial dilution method was used to detect the minimum inhibitory and bactericidal concentrations, and, finally, gas chromatography with mass-selective detection was performed to study the inhibition of E. coli's growth. To study the redox activity of different concentrations of nanocerium, we studied the intensity of chemiluminescence in the oxidation reaction of luminol in the presence of hydrogen peroxide. As a result of this study's use of the agar diffusion and serial dilution methods followed by sowing, no significant evidence of antimicrobial activity was found. At the same time, in the current study of antimicrobial activity against E. coli strains using gas chromatography with mass spectrometry, the ability of nanoceria to significantly inhibit the growth and reproduction of microorganisms after 24 h and, in particular, after 48 h of incubation at a wide range of concentrations, 10-2-10-5 M (48-95% reduction in the number of microbes with a significant dose-dependent effect) was determined as the optimum concentration. A reliable redox activity of nanoceria coated with citrate was established, increasing in proportion to the concentration, confirming the oxidative mechanism of the action of nanoceria. Thus, nanoceria have a dose-dependent bacteriostatic effect, which is most pronounced at concentrations of 10-2-10-3 M. Unlike the effects of classical antiseptics, the effect was manifested from 2 days and increased during the observation. To study the antimicrobial activity of nanomaterials, it is advisable not to use classical qualitative and semi-quantitative methods; rather, the employment of more accurate quantitative methods is advised, in particular, gas chromatography-mass spectrometry, during several days of incubation.

Growth performance, organs weight, intestinal histomorphology, and oocyst shedding in broiler chickens offered novel single strain Bacillus subtilis isolated from camel dung and challenged with Eimeria

We evaluated a single strain Bacillus subtilis BS-9 direct-fed microbial (BSDFM) isolated from camel dung in Eimeria challenged broiler chickens. Seven-hundred d-old Ross 708 male chicks were placed in pens (25 birds/pen) and allocated to 2 treatments (n = 14). From d 0 to 13, control pens received untreated water (-BSDFM), and 2 treated pens received water and 2 mL x 108 colony forming unit/bird/d (+BSDFM); daily water intake (WI) was recorded. On d 9, birds in half (+Eimeria) of pens per treatment received of 1 mL of Eimeria maxima and Eimeria acervulina oocysts orally, and the other half (-Eimeria) sterile saline solution. Birds had ad libitum access to feed and a water line from d 14. Feed intake (FI), body weight (BW) and mortality were recorded for calculating BW gain (BWG) and feed conversion ratio (FCR). On d 14 and 35, samples of birds were necropsied for organ weight and intestinal measurements. Excreta samples were collected from d 14 to 19 for oocyst count. There was no treatment effect (P > 0.05) on growth performance or WI on d 0 to 9. There were interactions between BSDFM and Eimeria on d 19 (P = 0.014) and 29 (P = 0.036) BW with unchallenged +BSDFM birds being heavier than birds in the other treatments. The main effects (P < 0.05) on d 10 to 35 FI, BW, and BWG were such that +BSDFM increased and Eimeria decreased (P < 0.01) these parameters. There was interaction (P = 0.022) between BSDFM and Eimeria on d 10 to 35 FCR such that the FCR of challenged -BSDFM birds was poor than that of unchallenged counterparts, but none differed with +BSDFM birds. There was an interaction (P = 0.039) between BSDFM and Eimeria on d 14 bursa weight with challenged birds exhibiting heavier bursa than unchallenged +BSDFM birds. Eimeria reduced (P = 0.01) and BSDFM (P = 0.002) increased the villi height to crypt depth ratio. Results showed that BSDFM supplementation via water can support the growth performance of broiler chickens challenged with Eimeria and may be a strategy to reduce adverse effects of coccidiosis.

Photocatalytic Degradation of Rhodamine-B and Water Densification via Eco-Friendly Synthesized Cr2O3 and Ag@Cr2O3 Using Garlic Peel Aqueous Extract

The purification and densification of wastewater play an important role in water recycling, especially if the materials used in water recycling are other types of recycled waste. Therefore, considering this view in this study, the biosynthesis of silver-decorated chromium oxide nanoparticles utilizing a wasted Allium sativum (garlic) peel extract is investigated. The aqueous extract of garlic peel (GPE) was treated with silver nitrate, chromium nitrate, and a mixture of silver nitrate and chromium nitrate to synthesize silver nanoparticles (Ag-garlic), chromium oxide nanoparticles (Cr2O3-garlic), and silver-decorated chromium oxide nanoparticles (Ag@Cr2O3-garlic), respectively. The synthesized nanoparticles were elucidated via thermal gravimetric analysis (TGA), infrared spectra (FT-IR), absorption spectra (UV-Vis), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). Antimicrobial activity studies were conducted against waterborne germs, bacterial strains (Bacillus subtilis, Enterococcus faecium, Escherichia coli, and Pseudomonas aeruginosa), and fungal strains (Alternaria porri, Aspergillus flavus, Aspergillus niger, Fuserium oxysporum, and Trichoderma longibrachiatum) and showed significant levels of antimicrobial activity. The results revealed that Ag@Cr2O3 significantly improved antimicrobial activity due to their synergistic effect. The photocatalytic activity of nanoparticles was assessed using Rhodamine B dye (5 ppm) under solar irradiation. Cr2O3-garlic exhibited the best activity as a photocatalyst among the studied nanoparticles, with 97.5% degradation efficiency under optimal conditions.

Characterization and Identification of Thermophilic Bacteria Isolated from Different Sites Located in Al-Jouf Region, Saudi Arabia

This study aims to investigate the morphological and biochemical characteristics of thermophilic bacteria isolated from thermophilic biotopes in the Al-Jouf region of Saudi Arabia for the first time. Additionally, the production of thermostable bacterial enzymes (proteases, amylases, cellulases, gelatinases, and lipases) was investigated. Six main bacterial species were identified via 16S rRNA sequencing and phylogenetic analysis. Twenty water and soil samples were collected from several thermophilic sites in the Dumat Al-Jandal and Al-Qurayyat cities. In total, 28 bacterial strains were isolated and biochemically characterized. Most isolated strains showed high protease and amylase production. 46 and 28% of isolated strains showed the production of cellulase and gelatinase, respectively. However, lipase production was not observed in any isolated bacteria. Four Bacillus licheniformis and two Bacillus subtilis strains exhibited high GC content (55%). Our findings suggest thermophilic bacteria as a source of thermostable enzymes for pharmaceutical and industrial applications.

Çeşitli Kaynaklardan ve Ticari Gıda Takviyelerinden İzole Edilen Bacillus Türlerinin Bazı Probiyotik Özelliklerinin Karşılaştırmalı Değerlendirilmesi

Bacillus species are gram-positive, aerobic, peritrically flagellated and endospore-forming bacteria. They can be found everywhere in the environment, especially in soil (its common habitat), water, dust or in the air. Probiotics, which have beneficial health effects, constitute an important group of Bacillus species. This study aimed to isolate Bacillus from various sources, identify it molecularly and determine its probiotic properties.For this purpose, eight Bacillus subtilis, Bacillus coagulans and Bacillus clausii strains among 58 isolates from fish intestine, soil, ripened cheese and commercial probiotic supplements were identified and their probiotic properties were characterized. Firstly, Bacillus strains were molecularly identified by 16S rRNA PCR analysis. The growth of Bacillus isolates at various temperatures, salt concentrations, and pH levels, as well as tests for esculin hydrolysis, starch hydrolysis, nitrate reduction, and gas generation from glucose, were all investigated to assess the isolates' physiological and biochemical characteristics. In terms of probiotic potential of Bacillus isolates; tolerance of bile salt, cell surface hydrophobicity, auto aggregation, antibiotic susceptibility tests were conducted. In all analyses, strains obtained from food supplements showed high levels of hydrophobicity and auto-aggregation properties, and the highest values following these strains were observed in Bacillus subtilis strains (F1 and S2) isolated from fish intestines and soil, respectively. All strains showed strong growth features in bile salt conditions. It has been determined that antibiotic sensitivity varies depending on the strain. Overall, high sensitivity to tetracycline has been observed. In summary, this study revealed the potential probiotic properties of Bacillus isolates obtained from different sources. The study also compared these probiotic properties with probiotic Bacillus strains isolated from food supplements.

Innovative utilization of harvested mushroom substrate for green synthesis of silver nanoparticles: A multi–response optimization approach

In this work, harvested mushroom substrate (HMS) has been explored for the first time through a comprehensive optimization study for the green synthesis of silver nanoparticles (AgNPs). A multiple response central composite design with three parameters: pH of the reaction mixture, temperature, and incubation period at three distinct levels was employed in the optimization study. The particle size of AgNPs, UV absorbance, and the percentage of Ag/Cl elemental ratio were considered as the response parameters. For each response variable examined the model used was found to be significant (P < 0.05). The ideal conditions were: pH 8.9, a temperature of 59.4 °C, and an incubation period of 48.5 h. The UV–visible spectra of AgNPs indicated that the absorption maxima for AgNP–3 were 414 nm, 420 for AgNPs–2, and 457 for AgNPs-1. The XRD analysis of AgNPs-3 and AgNPs-2 show a large diffraction peak at ∼38.2°, ∼44.2°, ∼64.4°, and ∼77.4°, respectively, which relate to the planes of polycrystalline face-centered cubic (fcc) silver. Additionally, the XRD result of AgNPs–1, reveals diffraction characteristics of AgCl planes (111, 200, 220, 311, 222, and 400). The TEM investigations indicated that the smallest particles were synthesized at pH 9 with average diameters of 35 ± 6 nm (AgNPs–3). The zeta potentials of the AgNPs are −36 (AgNPs–3), −28 (AgNPs–2), and −19 (AgNPs–1) mV, respectively. The distinct IR peak at 3400, 1634, and 1383 cm−1 indicated the typical vibration of phenols, proteins, and alkaloids, respectively. The AgNPs were further evaluated against gram (+) strain Bacillus subtilis (MTCC 736) and gram (−) strain Escherichia coli (MTCC 68). All of the NPs tested positive for antibacterial activity against both bacterial strains. The study makes a sustainable alternative to disposing of HMS to achieve the Sustainable Development Goals (SDGs).

Nitrate biodegradation by indigenous bacteria strain Bacillus subtilis obtained from eutrophic waters of Dal Lake Srinagar (India): Mechanisms and optimization

The presence of elevated nitrate concentrations in natural water bodies is a cause of significant concern, owing to its potential ecological and human health ramifications. Dal lake, a eutrophic lake situated in Srinagar, India, which bears a substantial burden of nutrient pollution stemming from various sources. To tackle high nitrate levels, an indigenous bacterial strain from Dal Lake, Srinagar, India, was used for its biodegradative abilities. To enhance biodegradation, the bacterial strain's efficiency was rigorously tested across various environmental conditions, including temperature, nitrate concentration, pH, contact time, and adsorbent quantity. The active isolate, identified through genetic analysis via 16S rRNA sequencing, was determined to be Bacillus subtilis ON358108. Fourier-transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), BET (Brunauer–Emmett–Teller) analysis (BET), XRD (X-ray diffraction) was employed.These methodologies were employed to scrutinize various aspects of the adsorbent, including surface area, pore volume, crystalline structure, composition, and internal structure were used in characterizing and elucidating the behaviour of functional groups engaged in the biodegradation process, both prior to and post-nitrate uptake. Adsorption mechanisms were established from experiments using the Langmuir, Freundlich, and Temkin models. We analyzed adsorption kinetics using pseudo-first- and pseudo-second-order models. To optimize the nitrate biodegradation process, we applied Response Surface Methodology (RSM) based on a central composite design approach. This approach successfully removed 91 % (±1.5) of the effluent's nitrate with Bacillus subtilis ON358108.Furthermore, our study demonstrated that the selected isotherm models fit the adsorption process in the following sequence: Langmuir > Temkin > Freundlich. In addition, thermodynamic study revealed the process is spontaneous and endothermic.

Toward Effects of Hydrophobicity on Biosurfactant Production by Bacillus subtilis Isolates from Crude-Oil-Exposed Environments

Background: Due to their structural features, biosurfactants reveal promising physicochemical properties, making them interesting for various applications in different fields, such as the food, cosmetics, agriculture, and bioremediation sectors. In particular, the bioproduction of surfactin, one of the most potent microbially synthesized biosurfactant molecules, is of great interest. However, since the wild-type productivities are comparably low, stimulatory environmental conditions have to be identified for improved bioproduction This study aims to find a correlation between the hydrophobicity and production of the biosurfactant surfactin by B. subtilis isolates from crude-oil-contaminated soil and water. Methods: The surfactin production yield was characterized in adapted batch cultivations using high-performance thin-layer liquid chromatography (HPTLC). Defined hydrophobic environmental conditions were achieved by supplementation with hexadecane or polystyrene beads, and the effects on biosurfactant production were measured. Adaptations at the protein level were analyzed using mass spectrometry measurements. Results: The correlation between hydrophobicity and surfactin production was characterized using Bacillus subtilis strains ZH1 and P7 isolated from crude-oil-contaminated soil and water. Since these isolates show the biodegradation of crude oil and hexadecane as hydrophobic substrates, respectively, a first-time approach, using polystyrene beads, was applied to provide a hydrophobic environment. Interestingly, contrary to popular opinion, reduced biosurfactant production was determined. Using mass spectrometric approaches, the physiological effects of co-cultivation and the cellular response at the protein level were investigated, resulting in altered quantities of stress proteins and proteins involved in the carbon metabolism counter to polystyrene beads. Conclusions: Contrary to common opinion, increasing hydrophobicity does not have a stimulating effect, and even reduces the effect on the bioproduction of surfactin as the main biosurfactant using selected B. subtilis strains.