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Simple Summary: A feather-degrading bacterium, Bacillus velezensis PN1, was isolated, and solid-state fermentation conditions were optimized. The resulting fermented feather meal (FFM) was evaluated for bioavailability in broilers. B. velezensis PN1 showed the highest feather degradation rate, with optimal fermentation achieved at 37 °C for 48–72 h. FFM2, produced with 65% moisture at 37 °C for 48 h, was compared with FFM1 (produced at 27 °C) and commercial hydrolyzed feather meal (HFM). FFM2 was found to be more suitable for large-scale production due to its amino acid profile and fermentation efficiency. In a broiler growth trial, diets containing 5% FFM2 showed no significant difference in body weight, feed conversion ratio, or performance efficiency compared to the 5% HFM group (p > 0.05). However, both FFM2 and HFM groups showed lower weight gain than the 5% fish meal (FM) group (p < 0.05). Without supplemental amino acids, growth performance did not differ between FFM2 and HFM groups (p > 0.05). In conclusion, FFM produced by B. velezensis PN1 can completely replace HFM when included at 5% in broiler diets. In this study, an effective feather-degrading bacterium was isolated and the solid-state fermentation condition for feather degradation was optimized. The resulting fermented feather meal (FFM) was evaluated for its bioavailability in broilers. Four Bacillus strains were examined for feather degradation rates, with Bacillus velezensis PN1 exhibiting the highest rate (83.24%, p < 0.05). A 3 × 3 × 3 factorial design was used to test substrate moisture content (45%, 55%, 65%), temperature (27 °C, 37 °C, 47 °C), and incubation time (24, 48, 72 h) for optimized fermentation conditions. In vitro pepsin digestibility (IVPD) revealed a significant interaction between temperature and time (p < 0.05), and the optimal performance was achieved at 37 °C for 72 h, followed by 37 °C for 48 h. Considering production time and cost, FFM2 (produced with 65% moisture at 37 °C for 48 h) was further compared with FFM1 (produced under the same conditions but at a lower temperature of 27 °C), and commercial hydrolyzed feather meal (HFM). IVPD did not differ significantly between FFM1, FFM2, and HFM, as they all showed significantly higher digestibility compared to raw feathers (RFs). FFM1, as well as FFM2, had significantly higher histidine and lysine concentrations than RF (p < 0.05), while FFM2 had a significantly lower cysteine content (p < 0.05). Based on fermentation conditions and amino acid composition, FFM2 was considered more suitable for large-scale production and was used in a broiler growth trial. The inclusion of 5% FFM2 in the broiler diet did not significantly affect body weight at 35 days compared to the 5% HFM group (p > 0.05), although both groups showed significantly lower weight gain than the 5% fish meal (FM) group (p < 0.05). The feed conversion ratio and performance efficiency factor did not differ significantly between the FFM2, HFM, and FM groups (p > 0.05). In groups fed diets without supplemental crystalline amino acids, growth performance did not significantly differ between the HFM and FFM2 groups (p > 0.05), although both performed significantly worse than groups with amino acid supplementation. In conclusion, FFM produced by B. velezensis PN1 through solid-state fermentation enhances feather bioavailability in poultry and can completely replace HFM when included at 5% in broiler diets, offering a potential sustainable alternative for poultry nutrition on a commercial scale. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the antimicrobial effect of direct cold atmospheric plasma (CAPP) treatment for pre-harvest application using four different crop species: Hordeum vulgare L. (barley), Triticum aestivum L. (wheat), Brassica napus L. (rapeseed) and Lupinus angustifolius L. (lupine). The model bacterium Bacillus atrophaeus served as a proxy for spore-forming plant pathogens on the seed surface. After semi-dry inoculation of spores onto the seeds, treatment with two different plasma sources, a volume-dielectric barrier discharge and a corona discharge, and different exposure times was carried out. Subsequently, recovery of viable spores from the seeds' surfaces was performed. Moreover, seed viability was determined based on maximum germination, as well as water contact angle as a measure for seed surface hydrophilicity. Direct CAPP treatment was efficient in reducing viable spores of B. atrophaeus with no significant differences between the plasma sources, reaching a mean inactivation of 1 log10 CFU/mL across all treatment times and crops species. Maximum germination of seeds was not negatively affected under any treatment condition. Seed hydrophilicity was increased for both plasma sources tested. Overall, this study provides valuable information on the efficiency of direct CAPP treatment of seeds with the purpose of seed hygienization with the premise of unaltered seed vitality and evaluates the potential application in comparison with previous investigated CAPP methods. [ABSTRACT FROM AUTHOR]

Hydroponic greenhouses and vertical farms provide an alternative crop production strategy in regions that experience low temperatures, suboptimal sunlight, or inadequate soil quality. However, hydroponic systems are soilless and, therefore, have vastly different bacterial microbiota than plants grown in soil. This review highlights some of the most prevalent plant growth-promoting bacteria (PGPB) and destructive phytopathogenic bacteria that dominate hydroponic systems. A complete understanding of which bacteria increase hydroponic crop yields and ways to mitigate crop loss from disease are critical to advancing microbiome research. The section focussing on plant growth-promoting bacteria highlights putative biological pathways for growth promotion and evidence of increased crop productivity in hydroponic systems by these organisms. Seven genera are examined in detail, including Pseudomonas, Bacillus, Azospirillum, Azotobacter, Rhizobium, Paenibacillus, and Paraburkholderia. In contrast, the review of hydroponic phytopathogens explores the mechanisms of disease, studies of disease incidence in greenhouse crops, and disease control strategies. Economically relevant diseases caused by Xanthomonas, Erwinia, Agrobacterium, Ralstonia, Clavibacter, Pectobacterium, and Pseudomonas are discussed. The conditions that make Pseudomonas both a friend and a foe, depending on the species, environment, and gene expression, provide insights into the complexity of plant–bacterial interactions. By amalgamating information on both beneficial and pathogenic bacteria in hydroponics, researchers and greenhouse growers can be better informed on how bacteria impact modern crop production systems. [ABSTRACT FROM AUTHOR]

Cannabis sativa L. has a variety of uses, including fiber production, food, oil, and medicine. In response to environmental concerns regarding chemical fertilizers, Bacillus velezensis S141 was examined as a plant-growth-promoting bacterium (PGPB) for cannabis. This study evaluated the effects of S141 on cannabis growth and utilized transcriptomic analysis to identify the responsive pathways. Inoculation with S141 significantly increased growth in laboratory and field environments, with most of the bacteria residing in the leaves, followed by the stems and roots, as determined by quantitative polymerase chain reaction (qPCR). Transcriptomic analysis revealed 976 differentially expressed genes. Upregulated genes were associated with metabolism, cellular processes, and catalytic activities, especially in the biosynthesis of phenylpropanoid, plant–pathogen interactions, and hormone signaling pathways. S141 mutants deficient in the production of auxin and cytokinin displayed reduced growth enhancement, which affirmed the roles of these hormones in cannabis development. These findings emphasize the potential of S141 as a sustainable growth promoter for cannabis and provide insights into the underlying pathways it influences. [ABSTRACT FROM AUTHOR]

Douchi is a kind of soybean-fermented food in China. To explore the common and differential compounds in different Douchi, Douchi was fermented by Aspergillus niger, Rhizopus arrhizus, and Bacillus circulans, respectively, and co-fermented by the three strains in this study. The common and characteristic flavor compounds and common and characteristic non-volatile components of different strains of fermented Douchi were explored through GC×GC-TOFMS and UPLC–Q-TOFMS omics analysis. The result suggested that Pyrazines, ketones, and alkenes such as tetramethyl-pyrazine, 2,5-dimethyl pyrazine, furaneol, 2,3-butanedione, gamma-terpinene might contribute to the basic flavor of the Douchi fermented by A. niger, R. arrhizus, and B. circulans. Peptides, amines, and flavonoids, such as N–acetylhistamine, 7,3′,4′–trihydroxyflavone, (3S,8As)-3-isobutylhexahydropyrrolo[1,2-a]pyrazine-1,4-dione might contribute to the basic function of the above three Douchi. The common metabolic pathways involved in the fermentation were isoflavonoid biosynthesis, flavonoid biosynthesis, etc. Ketones and esters such as 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one, 3-octanone, 5-methylfurfural and nonanal contributed to the unique flavor, while betaine, oleanolic acid, saikosaponin D and leucine might contribute to the unique function of A. niger fermented Douchi. Alkenes, pyrazine, and ketones such as α-terpinene, ethyl-pyrazine, dihydro-3-methyl-2(3H)-furanone, and linalool might contribute to unique flavor, while cordycepin, 2-Phenylacetamide might contributed to the unique function of R. arrhizus fermented Douchi. The unique flavor of B. circulans fermented Douchi might derived from ketones and esters such as 3-acetyl-2-butanone, 2-tridecanone, propionic acid-2-phenylethyl ester, while vitexin, astragalin, and phenethylamine might contribute to the unique function. Compared with single-strain fermented Douchi, the flavor substances and non-volatile components in multi-strain fermented Douchi were more abundant, such as hexadecanoic acid methyl ester, benzeneacetic acid ethyl ester, 9,12-octadecadienoic acid ethyl ester, nuciferine, and erucamide. It was speculated that there were common and differential substances in Douchi fermented by Aspergillus niger, Rhizopus arrhizus, and Bacillus circulans, which might contribute to the basic and unique flavor and function. Compared with single-strain fermented Douchi, the flavor substances and metabolites in multi-strain fermented Douchi were more abundant. This study provided a reference for the research of flavor and functional substances of Douchi. [ABSTRACT FROM AUTHOR]

Douchi has attracted attention for its unique taste and rich health functions. This study investigated the nutrition, flavor and correlation between the flavor and microorganisms of two traditional bacterial douchi from the province of Gansu in northwest China. The findings reveal significant variations in nutrition, flavor compounds, and the microbiota between Longnan and Qingyang douchi. Three dominant bacterial genera (Carnobacterium, Ignatzschineria, and Bacillus) and one dominant bacterial genus (Pichia) were found in the QY douchi, while four bacterial genera (Bacillus, Ignatzschineria, Proteus, and Providencia) and three fungal genera (Pichia, Candida, and Rhodosporidium) were dominant in samples of the LN douchi. For flavor substances, a total of 48 volatile components were detected in Longnan douchi and 41 in Qingyang douchi. Using the relative odor activity value (ROAV), we identified five key flavor compounds in Longnan douchi and four key flavor compounds in Qingyang douchi. The correlation analysis showed that there were certain positive or negative correlations between the key microorganisms and the flavor of the two traditional bacterial douchi. The results of this study can serve as a theoretical reference for improving the quality and flavor of traditional douchi. [ABSTRACT FROM AUTHOR]

Extended shelf-life (ESL) refrigerated ready-to-eat foods are thermally pasteurized to ensure food safety and stability. However, surviving psychrotrophic Bacillus cereus spores can still pose a challenge. Studies predicting their behavior often overlook sporulation conditions. This study investigated the effect of sporulation temperature on germination of three Bacillus weihenstephanensis strains in specific nutrients (inosine and/or amino acids) with or without prior heat activation (80 °C, 10 min). Sporulation temperature variably affected germination, with stronger effects in moderately responsive strains and nutrients. Heat activation strongly stimulated germination, particularly in nutrients with poorer responses, mitigating differences induced by sporulation temperature. The influence of sporulation temperature on germination and growth in an ESL matrix at refrigeration temperatures (4 °C or 8 °C) in vacuum packaging after heat activation or commercial pasteurization (90 °C, 10 min) was also studied. The latter treatment increased germination rates of surviving spores; however, some strains suffered damage and lost viability upon germination at 4 °C but recovered and grew at 8 °C. These findings highlight the need to account for variability in spore recovery and outgrowth during quantitative risk assessments for psychrotrophic B. cereus in ESL foods. [ABSTRACT FROM AUTHOR]

Arthrospira platensis (Ap) and Bacillus coagulans (Bc) have been successfully used to develop functional foods, but a combination of both regarding functional implications in nutritional value and antioxidant capacity has not been explored. This work aimed to develop an artisanal wheat flour pasta with egg using 5% A. platensis and 1% B. coagulans GBI 6068 (labeled as Bc+Ap). Uncooked pasta was characterized regarding nutritional value; furthermore, total phenolic content, antioxidant capacity by 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric reducing antioxidant power (FRAP), pigment content, colorimetry assay, textural profile analysis, buffering capacity, and probiotic viability were carried out on uncooked and cooked pasta to assess the changes induced by cooking. The Bc+Ap pasta showed enhanced nutritional value with a significant increase in protein content (30.61%). After cooking, the pasta showed increased phenolic content (14.22% mg GAE/g) and antioxidant capacity (55.59% µmol Trolox equivalents/g and 10.88% µmol Fe+2/g) for ABTS and FRAP, respectively, as well as pigment content (6.72 and 1.17 mg/100 g) for chlorophyll a+b and total carotenoids, respectively, but relative impacts on colorimetric parameters in contrast to control (wheat flour pasta). Furthermore, Bc+Ap showed improved firmness (59%, measured in g), buffer capacity (87.80% μmol H+(g × ΔpH)−1), and good probiotic viability (7.2 ± 0.17 log CFU/g) after the cooking process. [ABSTRACT FROM AUTHOR]

Trichloroethylene (TCE) is a prevalent groundwater contaminant detected worldwide, and microbes are sensitive indicators and initial responders to these chemical contaminants causing disturbances to their ecosystem. In this study, microbes isolated from San Marcos River sediment were screened for their TCE degradation potential. Among the twelve isolates (SAN1-12), five isolates demonstrated TCE degradation within 5 days at 25 °C and 40 mg/L of TCE concentration in the following order: SAN8 (87.56%), SAN1 (77.31%), SAN2 (76.58%), SAN3 (49.20%), and SAN7 (3.36%). On increasing the TCE concentration to 80 mg/L, the degradation efficiency of these isolates declined, although SAN8 remained the prominent TCE degrader with 75.67% degradation. The prominent TCE-degrading isolates were identified as Aeromonas sp. SAN1, Bacillus sp. SAN2, Gordonia sp. SAN3, and Bacillus proteolyticus SAN8 using 16S rRNA sequencing. The TCE degradation and cell biomass of Bacillus proteolyticus SAN8 were significantly improved when the incubation temperature was increased from 25 °C to 30 °C. However, both slightly acidic and alkaline pH levels, as well as higher TCE concentrations, lowered the efficacy of TCE degradation. Nevertheless, these conditions led to an increase in bacterial cell biomass. [ABSTRACT FROM AUTHOR]

Rice false smut, caused by the plant pathogenic fungus Ustilaginoidea virens, is widespread in rice-growing regions globally, severely compromising rice quality and production. Employing Bacillus spp. to control rice false smut represents an effective and environmentally sustainable strategy for disease management. The lipopeptides produced by Bacillus velezensis Jt84 demonstrated robust inhibitory effects against U. virens, resulting in abnormal mycelial morphology and spore germination. Iturins were identified as essential for the antifungal activity against U. virens, as confirmed by mutagenesis experiments that suppressed iturin biosynthesis. The surfactin-deficient mutant exhibited inhibitory effects against U. virens comparable to the wild-type, indicating that the absence of surfactins did not diminish its antifungal activity. Both the Jt84∆srf and Jt84∆itu mutants displayed reduced biofilm formation capabilities compared to the wild-type, with the Jt84∆srf mutant being particularly impaired and unable to form a complete biofilm. Regarding swarming motility, the ∆srf mutant exhibited a significant reduction compared to the wild-type, whereas the Jt84∆itu mutant showed a modest increase. Colonization experiments revealed that the Jt84∆srf mutant strain had significantly lower colonization on rice leaf surfaces than the wild-type strain, highlighting the critical role of surfactins in the colonization of B. velezensis Jt84 on rice leaves. In conclusion, our research demonstrated that surfactins and iturins have distinct functionalities and act synergistically to contribute to the biocontrol of rice false smut in B. velezensis Jt84. This synergy is achieved through their potent antifungal effects, biofilm formation, and successful colonization. [ABSTRACT FROM AUTHOR]

Forest cover is deteriorating rapidly due to anthropogenic causes, making its restoration urgent. Plant growth-promoting bacteria (PGPB) could offer a viable solution to ensure successful reforestation efforts. This study aimed to select bacterial strains with mechanisms that promote plant growth and enhance seedling development. The bacterial strains used in this study were isolated from the rhizosphere and endophyte regions of Pinus montezumae Lamb. and Pinus patula Schl. et Cham., two Mexican conifer species commonly used for reforestation purposes. Sixteen bacterial strains were selected for their ability to produce auxins, chitinase, and siderophores, perform nitrogen fixation, and solubilize inorganic phosphates; they also harbored genes encoding antimicrobial production and ACC deaminase. The adhesion to seeds, germination rate, and seedling response of P. montezumae and P. patula were performed following inoculation with 10 bacterial strains exhibiting high plant growth-promoting potential. Some strains demonstrated the capacity to enhance seedling growth. The selected strains were taxonomically characterized and belonged to the genus Serratia, Buttiauxella, and Bacillus. These strains exhibited at least two mechanisms of action, including the production of indole-3-acetic acid, biological nitrogen fixation, and phosphate solubilization, and could serve as potential alternatives for the reforestation of affected areas. [ABSTRACT FROM AUTHOR]

The current knowledge of Staphylococcus warneri phages is limited, with few genomes sequenced and characterized. In this study, a prophage, vB_G30_01, isolated from Staphylococcus warneri G30 was characterized and evaluated for its lysogenic host range. The phage was studied using transmission electron microscopy and a host range. The phage genome was sequenced and characterized in depth, including phylogenetic and taxonomic analyses. The linear dsDNA genome of vB_G30_01 contains 67 predicted open reading frames (ORFs), classifying it within Bronfenbrennervirinae. With a total of 10 ORFs involved in DNA replication-related and transcriptional regulator functions, vB_G30_01 may play a role in the genetics and transcription of a host. Additionally, vB_G30_01 possesses a complete set of genes related to host lysogeny and lysis, implying that vB_G30_01 may influence the survival and adaptation of its host. Furthermore, a comparative genomic analysis reveals that vB_G30_01 shares high genomic similarity with other Staphylococcus phages and is relatively closely related to those of Exiguobacterium and Bacillus, which, in combination with the cross-infection assay, suggests possible cross-species infection capabilities. This study enhances the understanding of Staphylococcus warneri prophages, providing insights into phage–host interactions and potential horizontal gene transfer. [ABSTRACT FROM AUTHOR]

In order to prevent structural damage or high repair costs caused by concrete crack propagation, the use of microbial-induced CaCO3 precipitation to repair concrete cracks has been a hot topic in recent years. However, due to environmental constraints such as oxygen concentration, the width and depth of repaired cracks are seriously insufficient, which affects the further development of microbial self-healing agents. In this paper, a ternary microbial self-healing agent composed of different proportions of Bacillus pasteurii, Saccharomyces cerevisiae, and Bacillus mucilaginosus was designed, and its crack repair ability was evaluated. When the mixing ratio was 7:1:2, the cell concentration was the highest, the precipitation amount of CaCO3 was the highest, and the crystallinity of calcite crystal was the highest. Compared to the single microorganism, the mortar specimens with ternary microorganisms had the largest repair area (up to 100%) and the deepest repair depth (CaCO3 presents at 9–12 mm from the crack surface). This is because when the concrete breaks, all three microorganisms are activated by water, O2, and CO2. Saccharomyces cerevisiae and Bacillus mucilaginosus accelerated the growth of Bacillus pasteurii and more mineralized products; CaCO3 was rapidly formed and quickly filled on the crack surface. When CaCO3 seals the surface of the crack, the internal Saccharomyces cerevisiae and Bacillus mucilaginosus continue to play a role. Bacillus mucilaginosus can accelerate the dissolution of CO2 produced by the anaerobic fermentation of Saccharomyces cerevisiae and the hydrolysis of CO32−, thereby improving the repair of the crack depth direction. [ABSTRACT FROM AUTHOR]

This study investigated the germination response to temperature of seeds of nine Arabidopsis thaliana ecotypes. They are characterized by a similar temperature dependency of seed germination, and 10 °C and 29 °C were found to be suboptimal low and high temperatures for all nine ecotypes, even though they originated from regions with diverse climates. We tested the potential of four PGPR strains from the genera Pseudomonas and Bacillus to stimulate seed germination in the two ecotypes under these suboptimal conditions. Biopriming of seeds with only the biofilm-forming strain Pseudomonas putida KT2440 significantly increased the germination of Cape Verde Islands (Cvi-0) seeds at 10 °C. However, biopriming did not significantly improve the germination of seeds of the widely utilized ecotype Columbia 0 (Col-0) at any of the two tested temperatures. To functionally investigate the role of KT2440's biofilm formation in the stimulation of seed germination, we used mutants with compromised biofilm-forming abilities. These bacterial mutants had a reduced ability to stimulate the germination of Cvi-0 seeds compared to wild-type KT2440, highlighting the importance of biofilm formation in promoting germination. These findings highlight the potential of PGPR-based biopriming for enhancing seed germination at low temperatures. [ABSTRACT FROM AUTHOR]

The crude metabolic extract from plant biocontrol bacteria plays a very important role in sustainable agricultural production. These extracts help maintain healthy plants and have very important application prospects in biotechnology related to alleviating apple replant disease (ARD). In this study, Bacillus velezensis XC1 (T1), Bacillus amyloliquefaciens QSB-6 (T2), and Lactobacillus reuteri LBR (T3) were examined to characterize the ability of their crude metabolic extracts to alleviate ARD. The high-throughput sequencing data of the soil microbial community structure were analyzed in relation to LBR crude metabolic extracts, and an extensive untargeted metabolomic analysis of UHPLC-Qex active components was performed. Active LC-MS/MS revealed that the main secondary metabolites involved in the biological control exerted by L. reuteri included 3-hydroxypropionaldehyde, extracellular polysaccharides (EPS), p-hydroxybenzoic acid, and azelaic acid. These crude metabolic extracts significantly inhibited the growth of soil pathogenic fungi, reduced the abundance of Fusarium, promoted the abundance of beneficial bacteria such as Pseudomonas, and optimized the soil microbial community structure. Improved modern extraction and purification technologies will be able to offer additional insights into the mechanism of action of these secondary metabolites and enable them to be used in biological preparations to prevent and control ARD in the future, as well as to allow harmful chemical fumigants to be discontinued. [ABSTRACT FROM AUTHOR]

Probiotics are live microbiota that can confer their hereditary health benefits upon the host. They can positively alter the diversity of the host's gut microbiota population. Bacillus clausii is a spore-producing potential probiotic. Its application in the food industry has been highly recommended. Clausii spores are greatly resistant to harsh food processing treatment (heat and industrial pressure) and they can maintain their physiological traits (acid, bile salt) inside the human gut. The utilization of spores can enhance the nutritional viability of livestock, as well as the functionality of on-shelf products. This contemporary review covers the great attributes of B. clausii, including physiological characteristics, modes of action, probiotics benefits, a safety assessment, and the bacteria food industry applications for the purposes of producing enhanced functional foods known as probiotic foods. To our knowledge, although B. clausii has been widely applied in food industry, the amount of the literature that is dedicated to its role in sustainable food production and addresses its potential sporogenous probiotic behavior is fewer as compared to non-sporogenous lactic acid bacteria (Lactobacillus spp. and Bifidobacterium spp.). In our review, we also discovered a lack of empirical evidence on the synbiotic and synergistic behavior of clausii in combination with other active nutrients. Therefore, our review suggests that more studies should be conducted on the potential probiotic contribution of B. clausii in sustainable food production and its synergistic and synbiotic behavior in conjunction with other active nutrients. [ABSTRACT FROM AUTHOR]

Background/Objectives: Research-engaged academic institutions offer the opportunity to couple undergraduate education/citizen science projects with antimicrobial biodiscovery research. Several initiatives reflecting this ethos have been reported internationally (e.g., Small World, Tiny Earth, MicroMundo, Antibiotics Unearthed). These programs target soil habitats due to their high microbial diversity and promote initial screening with non-selective, nutrient media such as tryptic soy agar (TSA). However, evaluation of published outputs to date indicates that isolate recovery on TSA is consistently dominated by the genera Bacillus, Pseudomonas, and Paenibacillus. In this study, we evaluated the potential of soil extract agar to enhance soil isolate diversity and antibiosis induction outcomes in our undergraduate Antibiotics Unearthed research program. Methods: We comparatively screened 229 isolates from woodland and garden soil samples on both tryptic soy agar (TSA) and soil extract agar (SEA) for antimicrobial activity against a panel of clinically relevant microbial pathogens. Results: On one or both media, 15 isolates were found to produce zones of clearing against respective pathogens. 16S rRNA gene sequencing linked the isolates with three genera: Streptomyces (7), Paenibacillus (6), and Pseudomonas (2). Six of the Streptomyces isolates and one Pseudomonas demonstrated antimicrobial activity when screened on SEA, with no activity on TSA. Furthermore, incorporation of the known secondary metabolite inducer N acetyl-glucosamine (20 mM) into SEA media altered the pathogen inhibition profiles of 14 isolates and resulted in broad-spectrum activity of one Streptomyces isolate, not observed on SEA alone. In conclusion, SEA was found to expand the diversity of culturable isolates from soil and specifically enhanced the recovery of members of the genus Streptomyces. SEA was also found to be a superior media for antibiosis induction among Streptomyces isolates when compared to TSA. It was noted that Paenibacillus isolates' antibiosis induction demonstrated a strain-specific response with respect to the growth media used. Conclusions: The authors propose SEA inclusion of in soil screening protocols as a cost-effective, complementary strategy to greatly enhance outcomes in undergraduate/citizen science-engaged antimicrobial biodiscovery initiatives. [ABSTRACT FROM AUTHOR]

Background: Beef burgers are perishable meat products, and to extend their shelf life, EU Regulation 1129/11 permits the use of certain additives. Objectives: However, given the concerns of health-conscious consumers and the potential toxicity of synthetic substances, this study aimed to explore the use of fennel waste extracts as natural preservatives. Methods: This study characterized the bioactive compounds (phenolic content), the antioxidant activity (ABTS+ and DPPH assay), and the antimicrobial properties (against Salmonella enterica serotype Enteritidis, Escherichia coli, Staphylococcus aureus, Bacillus cereusi, and Pseudomonas aeruginosa) of different fennel waste extracts (LF, liquid fraction; SF, solid fraction and PF, pellet fraction). Additionally, the potential use of the best fennel extract was evaluated for its impact on beef burger shelf life (up to 18 days at 4 ± 1 °C) in terms of microbiological profile, pH, and activity water (aw). Results: The PF extract, which was rich in flavones, hydroxybenzoic, and hydroxycinnamic acids, demonstrated the highest antioxidant and antimicrobial activities. Microbiological analyses on beef burgers with PF identified this extract as a potential antimicrobial substance. The aw and pH values did not appear to be affected. Conclusions: In conclusion, fennel extracts could be proposed as natural compounds exploitable in beef burgers to preserve their quality and extend their shelf-life. [ABSTRACT FROM AUTHOR]

This study explores sustainable methods for Sardine Processing Waste (SPW) valorization. Two approaches were investigated: (a) SPW microbial pretreatment adding Saccharomyces cerevisiae or Bacillus sp. in a two-stage anaerobic digestion (AD) for enzyme and biomethane production and (b) a single-stage AD without SPW pretreatment. Both S. cerevisiae and Bacillus sp. secreted proteases (0.66 and 0.58 U mL−1, respectively) and lipases (3.8 and 4.3 U mL−1, respectively) during hydrolysis, thus reducing viscosity (2.8 and 2.9 cP, respectively) compared with the untreated SPW (4.1 cP). Biomethane production was higher in the single-stage AD (1174 mL CH4 g−1 VS−1) when compared with the two-stage AD (821.5 and 260 mL CH4 g−1 VS−1 with S. cerevisiae and Bacillus sp., respectively). S. cerevisiae addition enhanced SPW degradation as implied by VS and sCOD values (70 and 84%, respectively), but this also resulted in a higher toxicity due to a three-fold increment in NH4-N content, reducing methanogen activity. This research demonstrates the innovative application of S. cerevisiae, a common bread-making yeast, in the biotechnological enhancement of SPW hydrolysis. Non-genetically engineered S. cerevisiae not only co-produced proteases and lipases but also significantly improved solubilization, degradation, and viscosity reduction, thereby rendering the yeast a key player in solid fish waste valorization, beyond its traditional applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Probiotics have a range of effects on poultry industry, including improving production performance, inhibiting or mitigating pathogenic infections, enhancing antioxidant capacity, reducing toxic effects, and improving the intestinal environment of poultry. They have significant potential for use as an ideal alternative to antibiotics in poultry farming. The purpose of this research was to evaluate the effect of solid fermentation products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans on the growth characteristics of broilers and the prevention of avian colibacillosis. The findings indicate that solid fermentation products can enhance the growth characteristics, immune function, and intestinal morphology of broilers. Additionally, it can optimize the abundance of microflora in the cecum and prevent avian colibacillosis, which is caused by avian pathogenic Escherichia coli (APEC). This study investigates the impact of the solid-state fermentation products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans (LCBs) on the growth characteristics, immune function, intestinal morphology, cecum microbial community, and prevention of avian colibacillosis in broilers. One hundred and twenty Hyland Brown broilers (aged one day) were divided randomly into three groups (four replicates of ten broilers per group). (1) The CON group was fed a basal diet. (2) The MOD group was fed a basal diet. On day 40, APEC strain SX02 (1.1 × 105 CFU/g) was administered to the breasts of chickens in this group. (3) The LCBs group was fed a basal diet supplemented with fermentation products (98.5% basal diet + 0.5% Lactobacillus plantarum and Candida utilis solid-state fermentation products + 1.0% Bacillus coagulans solid-state fermentation products). On day 40, the LCBs group received the same treatment as the MOD group. The experiment lasted 43 days. This study found that the average daily gain (ADG) of the LCBs group was significantly higher than that of the MOD group (p < 0.05), indicating that LCBs can significantly increase the ADG of broilers and improve the feed conversion ratio. Furthermore, compared to the MOD group, the heart bacterial load was significantly reduced in the LCBs group (p < 0.05), and the lesions less severe in the heart, liver, and jejunum were observed (p < 0.05). Additionally, the detection of intestinal flora showed a significant increase in the abundance of beneficial bacteria in the cecum of the LCBs group, while the number of Escherichia coli and Shigella decreased significantly. In conclusion, the solid fermentation of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans can improve the growth performance of broilers while also protecting against avian pathogenic Escherichia coli infection. This demonstrates the potential usefulness of these LCBs in feed production. [ABSTRACT FROM AUTHOR]

In this study, the nutrients, phytochemicals (including isoflavone and ginsenoside derivatives), and antioxidant activities of cheonggukjang with different ratios (0%, 2.5%, 5%, and 10%) of mountain-cultivated ginseng (MCG) were compared and analyzed using microorganisms isolated from traditional cheonggukjang. The IDCK 30 and IDCK 40 strains were confirmed as Bacillus licheniformis and Bacillus subtilis, respectively, based on morphological, biological, biochemical, and molecular genetic identification, as well as cell wall fatty acid composition. The contents of amino acids and fatty acids showed no significant difference in relation to the ratio of MCG. After fermentation, isoflavone glycoside (such as daidzin, glycitin, and genistin) contents decreased, while aglycone (daidzein, glycitein, and genistein) contents increased. However, total ginsenoside contents were higher according to the ratio of MCG. After fermentation, ginsenoside Rg2, F2, and protopanaxadiol contents of cheonggukjang decreased. Conversely, ginsenoside Rg3 (2.5%: 56.51 → 89.43 μg/g, 5.0%: 65.56 → 94.71 μg/g, and 10%: 96.05 → 166.90 μg/g) and compound K (2.5%: 28.54 → 69.43 μg/g, 5.0%: 41.63 → 150.72 μg/g, and 10%: 96.23 → 231.33 μg/g) increased. The total phenolic and total flavonoid contents were higher with increasing ratios of MCG and fermentation (fermented cheonggukjang with 10% MCG: 13.60 GAE and 1.87 RE mg/g). Additionally, radical scavenging activities and ferric reducing/antioxidant power were significantly increased in fermented cheonggukjang. This study demonstrates that the quality of cheonggukjang improved, and cheonggukjang with MCG as natural antioxidants may be useful in food and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Bradymonabacteria, as the representative of the facultative prey-dependent predators, were re-classified from the preceding Deltaproteobacteria into the phylum Myxococcota and proposed as a novel class named Bradymonadia. However, it was ambiguous whether their predatory pattern and properties were similar to those of the other myxobacterial predators. Therefore, the physiologic features were compared to determine the similarities and differences during the process of group attack and kin discrimination. Comparative genomic analyses were performed to conclude the core genome encoded commonly by bradymonabacteria, Myxococcia, and Polyangia. In conclusion, we proposed that bradymonabacteria have a predation pattern similar to the that of the representative of opportunistic predators like Myxococcus xanthus but with some subtle differences. Their predation was predicted to be initiated by the needle-less T3SS*, and the S-motility mediated by T4P also participated in the process. Meanwhile, their group attacks relied on cell contact and cell destiny. Inter-species (strains) kin discriminations occurred without the existence of T6SS. However, no extracellular lethal substance was detected in the fermentation liquor culture of bradymonabacteria, and the death of prey cells could only be observed when touched by their cells. Moreover, the prey-selective predation was observed when the predator encountered certain prey from Bacillus (G+), Algoriphagus (G−), and Nocardioides (G+). Bradymonabacteria can be regarded as a potential consumer and decomposer, and preying on many sea-dwelling or human pathogenic bacteria allows this group a broad application prospect in marine culture and clinical disease control. Our study will provide more evidence for its exploitations and applications. [ABSTRACT FROM AUTHOR]

A Planetary Atmospheric Chamber (PAC) was used to create simulations of interplanetary conditions to test the spore survival of three Bacillus spp. exposed to interacting conditions of vacuum (VAC), simulated solar heating (HEAT), and simulated solar ultraviolet irradiation (UV). Synergism was observed among the experimental factors for all three Bacillus spp. tested that suggested the increased lethality of HEAT and UV when concomitantly exposed to VAC. The most aggressive biocidal effects were observed for assays with VAC + HEAT + UV conditions run simultaneously over short time-steps. The results were used to predict the accumulation of extremely rapid Sterility Assurance Levels (SALs; def., −12 logs of bioburden reduction) measured in a few minutes to a few hours for external surfaces of interplanetary spacecraft. Furthermore, the results were extrapolated to predict that approx. 1 × 104 SAL exposures might be accumulated for external surfaces on the Europa Clipper spacecraft during a 3.5-year transit time between Venus (0.7 AU) and Mars (1.5 AU) during a series of Venus–Earth–Earth gravity assists (VEEGA trajectory) to Jovian space. The results are applicable to external spacecraft surfaces exposed to direct solar heating and UV irradiation during transits though the inner solar system. [ABSTRACT FROM AUTHOR]

Volatile organic compounds (VOCs) produced by Bacillus species exhibit biocontrol activity against fungal pathogens of fruits and vegetables. However, research on the effect of VOCs on Aspergillus flavus in stored grains is limited. This study aimed to investigate the effects of VOCs extracted from the strain R2, which was isolated from unhulled rice and identified as Bacillus paramycoides on A. flavus in vitro and unhulled rice. R2 VOCs effectively inhibited conidial germination and the hyphal growth of A. flavus in vitro. Moreover, R2 VOCs reduced the fungal population, aflatoxin B1 (AFB1) levels, and free fatty acid (FFA) value by 90.8%, 67%, and 38.7%, respectively, in unhulled rice. Eighteen R2 VOCs were identified using headspace solid-phase micro-extraction gas chromatography–mass spectrometry, and the individual activity of the VOCs against A. flavus was tested in vitro. Benzaldehyde (Ben) and 3,7-dimethyl-1-octanol (Dmo) showed strong inhibitory activities against A. flavus on PDA plates, with inhibition rates of 100% and 91.2%, respectively, at a concentration of 20 μL/dish. Ben at the concentration of 0.09 mg/mL, Dmo at the concentration of 0.07 mg/mL, or a mixture of both at halved concentrations could reduce the fungal population, AFB1 levels, and FFA content in unhulled rice. Our findings suggest that R2 VOCs are good alternatives to traditional chemical fumigants for suppressing A. flavus in stored grains. However, further research is necessary to establish the optimal fumigation concentration of these two components in unhulled rice. The impact of their residues on grain quality should be explored through sensory evaluation and nutritional analysis, and their safety to the environment and human body should be evaluated through safety assessment. [ABSTRACT FROM AUTHOR]

For years, the Bacillus genus has encompassed a heterogeneous group of individuals whose main common trait was their ability to sporulate in the presence of oxygen. This criterion has been revised, resulting in the reclassification of several species into new genera and to a redefinition of the characteristics of the members of this taxon. Currently, the species of the genus are grouped into the Subtilis clade and the Cereus clade. The former, called Bacillus sensu stricto, initially composed of B. subtilis, B. licheniformis, B. pumilus, and B. amyloliquefaciens, has subsequently incorporated new species related to these. The Cereus clade, Bacillus cereus sensu lato, consists of pathogenic species (B. anthracis, B. cereus, and B. thuringiensis) as well as others of significance in agriculture and industry. Furthermore, identifying these individuals remains complex, requiring alternatives to 16S rRNA sequencing. The ability to form spores resistant to stressful conditions provides a significant advantage over other genera, with observable differences in sporulation rates and spore structure among different species. Additionally, Bacillus spp. are known for their capacity to produce antimicrobial substances, lytic enzymes, and volatile organic compounds, each with diverse applications. Some species are even used as probiotics. This review delves into aspects related to the taxonomy and identification of microorganisms belonging to the genus Bacillus, which often present challenges. The aim is to provide a comprehensive overview of the topic. In addition, it highlights the characteristics and applications of the genus, emphasizing its importance in biotechnology and microbiology. [ABSTRACT FROM AUTHOR]

Sclerotinia minor (S. minor) Jagger is a phytopathogenic fungus that causes lettuce drop, a serious problem in lettuce (Lactuca sativa L.) production. The control of this pathogen is challenging because of the resistance of sclerotia, which can survive in the soil under favorable conditions. In Bulgaria, the management of lettuce drop relies primarily on the strategic application of synthetic fungicides. To find alternative methods for disease management, four bacterial isolates were screened for antagonism against S. minor. This study reports the in vitro evaluation of the antifungal activity of Bacillus subtilis, Priestia megaterium, Bacillus safensis, and Bacillus mojavensis against S. minor. The molecular identification of the isolates involved in the activity was examined through 16s rRNA sequencing. Isolated bacterial strains produced indole-3-acetic acid (IAA) in a medium supplemented with 0.1% L-tryptophan. The ability of these strains to increase the mobility of phosphorus and zinc was elucidated. The production of siderophores was confirmed on CAS (Chrom azurol S) medium. The inhibitory action of the bacterial growth broth filtrates against S. minor was demonstrated, indicating the nature of the molecules involved. The evaluation of antifungal activity was carried out in vitro and in pot experiments. This study determined the effect of growth-promoting rhizobacteria on the development of lettuce. This research focuses mainly on the development of biocontrol strategies for the management of lettuce drop in greenhouses. [ABSTRACT FROM AUTHOR]

Seven auxin-producing endophytic bacterial strains (Azospirillum spp., Methylobacterium symbioticum, Bacillus spp.), and two different combinations of these strains were used to verify their influence on tomato during germination and development in hydroponic conditions where, as a novelty for Canestrino di Lucca cultivar, endophytic bacteria were inoculated. To emphasize the presence of bacterial auxins in roots and stems of seedlings, both in situ staining qualitative assessment and quantitative analysis were carried out. Moreover, hypogeal and epigeal growth of the plantlets were measured, and correlation analyses were conducted to examine the relationship between the amount of indolacetic acid (IAA) produced by the bacterial strains and root and stem parameters. Plantlets treated with microbial inoculants showed a significant increase in the survival rate compared to the control treatment. The best results as IAA producers were from Azospirillum baldaniorum Sp245 and A. brasilense Cd, which also induced significant root growth. On the other hand, Bacillus amyloliquefaciens and B. licheniformis induced the best rates in stem growth. These findings highlight the potential for using endophytic bacterial strains in a hydroponic co-cultivation system that enables inoculating plantlets, at an early stage of growth (5 days old). [ABSTRACT FROM AUTHOR]

Fusarium pseudograminearum (Fpg) is a significant pathogen responsible for fusarium crown rot (FCR) in wheat (Triticum aestivum L.), a disease with devastating impacts on crop yield. The utilization of biocontrol bacteria to combat fungal diseases in plants is a cost-effective, eco-friendly, and sustainable strategy. In this trial, an endophytic bacterial species, designated as SW, was isolated from the roots of wheat. The strain exhibited potent antagonistic effects against Fpg and reduced the FCR disease severity index by 76.07 ± 0.33% in a greenhouse pot trial. Here, 106 colony-forming units (CFUs)/mL of the SW strain was determined to be the minimum dose required to exhibit the antagonism against Fpg. The strain was identified as Bacillus atrophaeus using genome sequencing and comparison with type strains in the NCBI database. Whole-genome sequencing analysis revealed that SW harbors genes for siderophores, antifungal metabolites, and antibiotics, which are key contributors to its antagonistic activity. Additionally, the strain's ability to utilize various carbon and nitrogen sources, successfully colonize wheat root tissues as an endophyte, and form biofilms are critical attributes for promoting plant growth. In summary, these findings demonstrate the ability of Bacillus atrophaeus to control FCR disease in wheat in a sustainable agricultural setting. [ABSTRACT FROM AUTHOR]

YqeK is a bacterial HD-domain metalloprotein that hydrolyzes the putative second messenger diadenosine tetraphosphate (Ap4A). Elevated Ap4A levels are primarily observed upon exposure of bacteria to factors such as heat or oxidative stress and cause pleiotropic effects, including antibiotic sensitivity and disrupted biofilm formation. Ap4A thus plays a central role in bacterial physiology and metabolism, and its hydrolysis by YqeK is intimately linked to the ability of these microbes to cope with stress. Although YqeK is reported to hydrolyze Ap4A under aerobic conditions, all four existing crystal structures reveal an active site that consists of a diiron center, portraying a cryptic chemical nature for the active metallocofactor. This study examines two YqeK proteins from two ecologically diverse parent organisms: the obligate anaerobe Clostridium acetobutylicum and the facultative aerobe Bacillus halodurans. Both enzymes utilize Fe-based cofactors for catalysis, while under ambient or oxidative conditions, Bh YqeK hydrolyzes Ap4A more efficiently compared to Ca YqeK. This redox-dependent activity difference stems from the following two molecular mechanisms: the incorporation of mixed-metal, Fe-based bimetallic cofactors, in which the second metal is redox inert (i.e., Fe–Zn) and the upshift of the Fe–Fe cofactor reduction potentials. In addition, three strictly conserved, positively charged residues vicinal to the active site are critical for tuning Ap4A hydrolysis. In conclusion, YqeK is an Fe-dependent phosphohydrolase that appears to have evolved to permit Ap4A hydrolysis under different environmental niches (aerobic vs. anaerobic) by expanding its cofactor configuration and O2 tolerance. [ABSTRACT FROM AUTHOR]

The formation of cross-linked enzyme aggregates (CLEAs) using macromolecular cross-linkers improves substrate accessibility and enhances enzyme retention. However, there have been few studies exploring the use of macromolecular cross-linkers due to the challenges related to cross-linker screening. In compliance with our previous computational and experimental screening, dextran is the optimal macromolecular cross-linker to develop CLEAs of endolevanase from Bacillus lehensis G1 (rlevblg1-dex-CLEA) for levan-type-fructooligosaccharides (L-FOS) production. In this study, rlevblg1-dex-CLEAs was optimized, and the activity recovery continued to increase and reached 90.5%. Subsequently, the rlevblg1-dex-CLEAs were characterized and they displayed higher thermal stability after 1 h of incubation in comparison to the free enzyme. Moreover, the rlevblg1-dex-CLEAs were reusable for five cycles and exhibited greater storage stability over 180 days at 4 °C (60.9%) than that of free rlevblg1. In addition, the rlevblg1-dex-CLEAs demonstrated similar catalytic efficiency as the free enzyme and generated a substantial amount of L-FOS with a longer degree of polymerization, which is more beneficial for industrial use. [ABSTRACT FROM AUTHOR]

This study identified a salt-tolerant GH11 xylanase, Xynst, which was isolated from a soil bacterium Bacillus sp. SC1 and can resist as high as 4 M NaCl. After rational design and high-throughput screening of site-directed mutant libraries, a double mutant W6F/Q7H with a 244% increase in catalytic activity and a 10 °C increment in optimal temperature was obtained. Both Xynst and W6F/Q7H xylanases were stimulated by high concentrations of salts. In particular, the activity of W6F/Q7H was more than eight times that of Xynst in the presence of 2 M NaCl at 65 °C. Kinetic parameters indicated they have the highest affinity for beechwood xylan (Km = 0.30 mg mL−1 for Xynst and 0.18 mg mL−1 for W6F/Q7H), and W6F/Q7H has very high catalytic efficiency (Kcat/Km = 15483.33 mL mg−1 s−1). Molecular dynamic simulation suggested that W6F/Q7H has a more compact overall structure, improved rigidity of the active pocket edge, and a flexible upper-end alpha helix. Hydrolysis of different xylans by W6F/Q7H released more xylooligosaccharides and yielded higher proportions of xylobiose and xylotriose than Xynst did. The conversion efficiencies of Xynst and W6F/Q7H on all tested xylans exceeded 20%, suggesting potential applications in the agricultural and food industries. [ABSTRACT FROM AUTHOR]

Simple Summary: Comparing different commercial biofertilizer products for their effects as biostimulants, as protectors of plants from drought and plant-pathogenic fungi, can be difficult due to the increasing number of bacteria available in the market. Using a panel of tests, we compared the effects of four commercial strains of the operational group Bacillus amyloliquefaciens to assess their ability to protect pepper plants and stimulate their growth. Then, we obtained numerical values that allow quick diagnoses when choosing a product to ensure that it has the greatest added value or when describing a new strain. The market for bacteria as agricultural biofertilizers is growing rapidly, offering plant-growth stimulants; biofungicides; and, more recently, protectors against extreme environmental factors, such as drought. This abundance makes it challenging for the end user to decide on the product to use. In this work, we describe the isolation of a strain of Bacillus velezensis (belonging to the operational group Bacillus amyloliquefaciens) for use as a plant-growth-promoting rhizobacterium, a biofungicide, and a protector against drought. To compare its effectiveness with other commercial strains of the same operational group, Bacillus amyloliquefaciens, we analyzed its ability to promote the growth of pepper plants and protect them against drought, as well as its fungicidal activity through antibiosis and antagonism tests, its ability to solubilize potassium and phosphates, and its ability to produce siderophores. Finally, we used a probit function, a type of regression analysis used to model the outcomes of analyses, to quantify the biostimulatory effectiveness of the different plant-growth-promoting rhizobacteria, developing what we have called the Agricultural Protection Against Stress Index, which allowed us to numerically compare the four commercial strains of the operational group Bacillus amyloliquefaciens, based on a Delphi method—a type of regression analysis that can be used to model a cumulative normal distribution—and integrate the results from our panel of tests into a single value. [ABSTRACT FROM AUTHOR]

Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this goal. Unfortunately, the adaptability of insects has led to resistance to certain proteins in current commercial products. Therefore, new insecticidal traits representing a different mode of action (MoA) than those currently in use are needed. Cry1Ja has good insecticidal activity against various lepidopteran species, and it provides robust protection against insect feeding with in planta expression. For Bt proteins, different MoAs are determined by their binding sites in the insect midgut. In this study, competitive binding assays are performed using brush border membrane vesicles (BBMVs) from Helicoverpa zea, Spodoptera frugiperda, and Chrysodeixis includens to evaluate the MoA of Cry1Ja relative to representatives of the various Bt proteins that are expressed in current commercial products for lepidopteran insect protection. This study highlights differences in the shared Cry protein binding sites in three insect species, Cry1Ja bioactivity against Cry1Fa resistant FAW, and in planta efficacy against target pests. These data illustrate the potential of Cry1Ja for new insect trait development. [ABSTRACT FROM AUTHOR]

Butirosins are naturally occurring aminoglycoside (AG) antibiotics featuring a 4,5-disubstituted 2-deoxystreptamine (2-DOS) with a (2S)-4-amino-2-hydroxybutyrate (AHBA) side chain. This side chain has been shown to confer resistance against AG-modifying enzymes, leading to ongoing studies on the butirosin biosynthetic pathway and the corresponding enzymes. Butirosin is produced by Niallia (formerly Bacillus) circulans and Bacillus vitellinus, with most research focused on the first strain. To date, no whole-genome analysis has been performed on B. vitellinus. In this study, we sequenced the complete genome of B. vitellinus NBRC 13296 and performed a comparative analysis of different butirosin biosyntheric gene clusters (BGCs), including those from N. circulans. The complete genome of B. vitellinus NBRC 13296 comprises a 6,331,192-base circular chromosome with GC content of 52.68%. The annotation revealed the presence of 5605 CDSs, 70 tRNA genes, 30 rRNA genes, and 3 ncRNA genes in NBRC 13296. The highest dDDH and ANI values between NBRC 13296 and the most closely related type strain, Paenibacillus chitinolyticus KCCM 41,400, were 97.8% and 98.66%, respectively. Based on these genome-based comparative analyses, we propose reclassifying B. vitellinus NBRC 13296 as P. chitinolyticus. Genome mining revealed 18 gene clusters encoding the biosynthesis of diverse secondary metabolites in the genome of B. vitellinus NBRC 13296, indicating the enormous biosynthetic potential of this strain. The predicted structural diversity of the secondary metabolites includes aminoglycosides, PKS, NRPS, PKS–NRPS hybrids, metallophores, phosphonates, terpenes, β-lactones, and RiPP peptides. We then comparatively characterized the butirosin BGCs previously studied in several N. circulans strains. Additionally, the comparative genome analysis revealed complete butirosin BGCs identified from P. chitinolyticus KCCM 41,400, P. chitinolyticus NRRL B-23119, P. chitinolyticus NRRL B-23120, P. chitinolyticus B-14908, P. chitinolyticus YSY-3.1, P. chitinolyticus JMW06, Paenibacillus sp. GbtcB18, Paenibacillus sp. HGH0039, and Paenibacillus sp. MZ04-78.2. Finally, we identified the core region consisting of BtrS, BtrN, BtrM, BtrL, BtrA, BtrB, BtrC, BtrD, BtrD, BtrE, BtrF, BtrG, BtrH, BtrI, BtrI, BtrJ, BtrK, BtrO, BtrP, and BtrV, followed by an upstream region organizing BtrQ, BtrW, BtrX, BtrY, and BtrZ in the same transcriptional direction and sequential genetic arrangement, and a downstream region organizing various proteins based on BtrT, BtrR2, BtrU, and BtrR1. Our study provides insights into the reclassification of B. vitellinus NBRC 13296 to P. chitinolyticus and suggests the need for continued studies on butirosin biosynthesis from an enzymatic perspective. [ABSTRACT FROM AUTHOR]

The bacterial communities in rhizosphere soil interact with the roots of plants. This interaction is beneficial for both the bacteria and the plants, which makes it very important to identify the structure of these bacterial communities for plant growth and development. However, the composition characteristics of bacterial communities in rhizosphere soil of 2-year and 5-year Haloxylon ammodendron have not been clearly defined. The purpose of this study was to identify the diverse composition of 2-year and 5-year Haloxylon ammodendron in Turpan, Xinjiang. Thus, rhizosphere soil bacteria were analyzed by isolating, purifying, and identifying the species through high-throughput sequencing technology. The bacterial strains in the rhizosphere soil of Haloxylon ammodendron were isolated with the dilution coating method, resulting in 37 isolated strains. The selective media were used to screen the growth-promoting characteristics of the rhizosphere soil isolates of Haloxylon ammodendron. The results of high-throughput amplification sequencing showed that the rhizosphere bacteria in the 2-year rhizosphere soil belonged to 45 phyla, 109 classes, 288 orders, 451 families, 826 genera, and 404 species, and those in the 5-year rhizosphere soil belonged to 56 phyla, 148 classes, 369 orders, 601 families, 1062 genera, and 671 species. Among them, Firmicutes, Proteobacteria, Actinobacteriota, Bacteroidota, Crenarchaeota, and so on are the dominant bacteria. There were 12206 and 14,053 OTUs in the 2-year-old and 5-year-old rhizosphere soil bacteria, respectively, and 3329 OTUs in the 2-year- and 5-year-old rhizosphere soil, accounting for 16.98% of the total number of OTUs. The results showed that three strains, sg16, sg21, and ss4, had the highest inorganic phosphorus solubility index (1.58). The isolated strain did not have the ability to dissolve organophosphorus and potassium, while the screened strain sg16 had the ability to fix nitrogen. Two strains with a good iron-bearing capacity, Sg9F and Sg1, were screened, among which Sg9F had the highest D/d value and Sg9F had the strongest iron-bearing capacity. The results showed that 37 strains of rhizosphere soil bacteria belonged to six genera. They are Bacillus, Corynebacterium, Phyllobacterium, Lysinibacillus, Sinorhizobium meliloti, and Streptomyces levis. Among them are sg21 (Bacillus sp.), sg1 (Bacillus sp.), sg9F (Streptomyces levis), sg16 (Phyllobacterium phragmitis), and ss4 (Sinorhizobium meliloti). This study provides a particular research basis for the influence of Haloxylon ammodendron rhizosphere bacteria on soil nutrient release and depicts a solution for improving the yield and quality of cistanche deserticola indirectly through isolating, screening, and identifying rhizosphere soil bacteria, including screening strains with growth-promoting functions and analyzing the population structure of rhizosphere bacteria in 2- and 5-year soil in combination with high-throughput sequencing technology. [ABSTRACT FROM AUTHOR]

Drinking water sources with groundwater arsenic (As) contamination face multifaceted challenges in the removal and supply of fresh drinking water resources. To eradicate this problem, bioremediation has evolved to become more effective than other chemical and physical removal processes in its cost-effectiveness, high removal efficiency, and lesser production of secondary by-products or waste. Thus, this study aimed to treat As from aqueous media and to detoxify highly toxic forms of As by the isolated bacteria from As-affected areas. We isolated two new Gram-positive bacteria, which are reported here (Bacillus sp. and Bacillus cereus), with As5+ minimum inhibitory concentrations (MICs) of 4500 mg/L for the Bacillus sp. and 1000 mg/L for Bacillus cereus; meanwhile, for As3+, the MICs are 600 mg/L for both isolates. Bacillus sp. and Bacillus cereus can also effectively convert the highly toxic and easily mobile As3+ to As5+ in aqueous media. This study also demonstrates that these bacteria can remove a significant proportion of As3+ and As5+ (averaging 50% for both) from aqueous media. These As-resistant bacteria from the As-affected area can be used and upscaled for the treatment of As for a safer drinking water supply. [ABSTRACT FROM AUTHOR]

Ectoine is a compatible solute naturally produced in some halophilic bacteria as a protective agent for survival in salty environments. It has gained special interest as a therapeutic agent in the pharmaceutical and healthcare sectors for the treatment of different diseases. Ectoine mainly produced by bacterial milking, chemical, and fed-batch fermentation methods under a high-salt medium. Unfortunately, the ectoine yield through these methods is still too low to meet high industrial demand, causing salinity issues. The biobrick method was potentially utilized for efficient ectoine biosynthesis under a low-salt medium with different conditions in E. coli BL21(DE3) harboring the pET-22bNS-EctA-EctB-EctC plasmid. Firstly, three genes, L-2,4-diamino-butyric acid acetyltransferase (ectA), L-2,4-diaminobutyric acid transaminase (ectB), and ectoine synthase (ectC) from Bacillus pseudofirmus OF4, were precisely assembled and expressed into E. coli BL21(DE3). After optimizing the reaction conditions in a whole-cell catalytic reaction [50 mM of the sodium phosphate buffer (pH~7.5) containing 300 mM L-aspartic acid, 100 mM glycerol, 1/20 g/mL cell pellets], the amount of ectoine in the plasmid pET-22bNS-ALacBTacCTac reached the maximum level of 167.2 mg/mL/d (6.97 mg/mL/h). Moreover, Western blot analysis revealed that high expression levels of EctA and EctC had a significant effect on ectoine biosynthesis, indicating that both proteins might be the key enzymes in ectoine production. We conclude that a high amount of ectoine achieved through the biobrick method and efficiently used for different industrial applications. [ABSTRACT FROM AUTHOR]

The purpose of this study was to isolate, identify, and evaluate the antibacterial and probiotic potential of bacteria from honeydew honey collected in Poland. Isolates (189 colonies from 10 honey samples) were evaluated for their antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Yersinia enterocolitica, and then identified by MALDI-TOF-MS. The isolates with the greatest antimicrobial properties were screened for their probiotic potential. The total number of bacteria isolated from honey did not exceed the value of 2.5 × 102 CFU/mL. The Bacillus pumilus/altitudinis, B. licheniformis, and Bacillus cereus groups were the dominant identified bacteria. Almost 16% of the isolates expressed antibacterial potential against three pathogenic bacteria, over 20% against two, while almost 34% of the isolates did not inhibit any. The survival rate of the isolates under gastrointestinal tract conditions was higher after 4 h of exposure to bile salts (>60% survival rate for 66.66% of the isolates), while at pH 2.0, it was lower (>50% survival rate for 44% of the isolates). The most resistant isolate B. pumilus/altitudinis survived at a rate of 77% at low pH and 108% with bile salts. These results confirmed that honeydew honey is a promising reservoir of bacteria that produces metabolites with antimicrobial and probiotic potential. [ABSTRACT FROM AUTHOR]

The rapid emergence and spread of multidrug-resistant microorganisms is threatening our ability to treat common infections, with serious medical, social, and economic consequences. Despite substantial progress in the global fight against antibiotic resistance, the number of effective antibiotics is rapidly decreasing, underlying the urgent need to develop novel antimicrobials. In the present study, the green synthesis of novel iodine-substituted tricyclic flavonoids has been accomplished using an eco-friendly reagent, HPW-SiO2, as a cyclization agent for the precursor 3-dithiocarmamic flavanones. In vitro antimicrobial activity of the new compounds was evaluated using minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentrations. All tested compounds displayed potent inhibitory activity against all tested microbial strains, with the lowest MIC values of 0.12 µg/mL and 0.48 µg/mL recorded for compound 5c against Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus. Higher MIC values (7.81 µg/mL) were registered for several flavonoids against Gram-negative bacteria Escherichia coli and Acinetobacter pittii. No inhibitory activity was evidenced against Pseudomonas aeruginosa strain. The highest antifungal activity was displayed by flavonoid 5d against Candida krusei (MIC = 3.9 µg/mL). The same compound also exhibited the most potent bactericidal and fungicidal activity against Bacillus subtilis (0.9 µg/mL) and Staphylococcus aureus (1.97 µg/mL), Candida albicans, and Candida krusei (7.81 µg/mL). Based on the reported results, we believe that the novel iodine-substituted tricyclic flavonoids have good potential to become new antimicrobial agents effective against bacterial and fungal strains, including WHO-priority pathogens. [ABSTRACT FROM AUTHOR]

Jeju, the largest island in Korea, is the most economically important in terms of marine aquaculture. We investigated the marine viral composition adjacent to Jeju Island over four seasons in 2022 and sequenced DNA libraries extracted from samples in March, June, September, and December using Illumina HiSeq 2000. We obtained 212,402, 186,542, 235,441, and 224,513 contigs from the four-season samples, respectively. Among the identified metagenomes, bacteriophages were dominant in all the samples. Bacillus phage G was the dominant species in March and June, whereas Pelagibacter phage HTVC 008M was the dominant species in September and December. Additionally, the number of viruses that infected algal hosts was higher in December than in other seasons. Marine viruses appeared in all seasons and infected marine vertebrates such as fish. Functional analysis using MG-RAST revealed that cell wall- and capsule-related metabolism groups were activated in March and June, whereas virulence-, disease-, and defense-related metabolism groups were activated in September and December. Conclusively, this study revealed seasonal changes in marine viral communities in the sea adjacent to Jeju Island. Our data will be useful in identifying emerging marine viral pathogens and for further community studies on marine organisms. [ABSTRACT FROM AUTHOR]

In this study, the effects of adding glutamate (Glu), glutamine (Gln), aspartate (Asp), and asparagine (Asn) on the flavor formation of Huangjiu were investigated, and the effect of Gln concentration on the quality, microbial community structure, and flavor development of Huangjiu was further explored. Varied Gln concentrations influenced yeast growth, sugar utilization, microbial communities, and quality attributes. Additional Gln promoted yeast cell counts and sugar depletion. It increased the complexity of bacterial co-occurrence networks and reduced the impact of stochastic processes on assembly. Correlation analysis linked microorganisms to flavor compounds. Isolation experiments verified the role of Saccharomyces cerevisiae, Aspergillus chevalieri, Bacillus altitudinis, and Lactobacillus coryniformis in flavor production under Gln conditions. This research elucidated the microbiological mechanisms by which amino acid supplementation, especially Gln, enhances Huangjiu quality by modulating microbial metabolic functions and community dynamics during fermentation. This research is significant for guiding the production of Huangjiu and enhancing its quality. [ABSTRACT FROM AUTHOR]

Vanilla planifolia is grown as a high-value orchid spice for its odor and savor attributes that increase due to the curing process associated with microbial colonization. This tends to influence the aromatic properties of vanilla. Hence, 11 Bacillus sp. strains were isolated from V. planifolia and identified with 16S rRNA gene sequencing. The liquid culture (1 mL of 107 CFU mL−1) of selected Bacillus vallismortis NR_104873.1:11-1518, Bacillus velezensis ZN-S10, and Bacillus tropicus KhEp-2 effectively fermented green-blanched vanilla pods kept at 10 °C during the sweating stage. GC-MS analysis showed that the methanol extract of non-coated, and B. vallismortis treated vanilla detected three (3) volatile compounds, whereas seven (7) components were obtained in B. tropicus and B. velezensis treatment. 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl was found in B. velezensis ZN-S10, B. tropicus KhEp-2, and B. vallismortis while it was not present in the control samples. This ketone compound suggested a Maillard reaction resulting in brown-increased aroma pods. Linoleic acid and Hexadecanoic acid ethyl esters were detected only in ZN-S10 strain-coated vanilla. A novel 3-Deoxy-d-mannoic lactone was detected only in B. vallismortis-treated vanilla characterized as a new compound in V. planifolia which suggested that the new compound can be altered with the coating of bacteria in vanilla during fermentation. Thus, the Bacillus strains improved the volatile profile and exhibited a new aroma and flavor profile of vanilla owing to bacteria fermentation during the curing process. [ABSTRACT FROM AUTHOR]

Mycotoxins, toxic compounds produced by fungi, pose significant risks to food safety and human health. This study investigates the bio-detoxification potential of 238 strains of lactic acid bacteria (LAB) and Bacillus spp., previously isolated from cereals (including mycotoxin-contaminated grains), against the emerging mycotoxin, enniatin B (ENB), and the prevalent mycotoxin, deoxynivalenol (DON). Out of the tested strains, 26 demonstrated notable mycotoxin reduction capabilities, including 2 Bacillus pumilus and 24 Bacillus licheniformis strains. B. licheniformis strains MA572, MA695, MA696, TR174a, TR284, TR363, and TR466a degraded ENB to levels below the detection limit, and six strains reduced DON by 30–35%; B. licheniformis TR251b and TR374 showed the highest DON reduction with 35.7%. The most promising strains for bio-detoxification were B. licheniformis TR284, which achieved a 100% reduction in ENB and a 28.6% reduction in DON and B. licheniformis TR388 with a 97.5% reduction in ENB and a 31.9% reduction in DON. None of the tested LAB strains significantly reduced either mycotoxin. These findings highlight the promising potential of B. licheniformis strains in bio-detoxifying mycotoxin-contaminated cereal products. Further research into the underlying detoxification mechanisms and safety aspects is essential to develop effective bio-detoxification strategies for enhancing food safety. [ABSTRACT FROM AUTHOR]

It was previously discovered that, in the Gram-negative bacterium Escherichia coli growing on a minimal medium with sulfate, stress-induced growth arrest is accompanied by the release of hydrogen sulfide. The source of the sulfide is the desulfurization of intracellular cysteine as one of the ways of maintaining it at a safe level. The danger of excess cysteine is associated with its participation in the Fenton reaction, leading to the formation of highly toxic hydroxyl radicals. Using electrochemical sensors, we identified stress-induced sulfide production in the Gram-positive bacteria Bacillus subtilis and Bacillus megaterium, growing on a minimal medium with sulfate, and changes in physiological parameters such as Eh, pH, and oxygen and potassium consumption. Sulfide production was observed during growth arrest due to the depletion of glucose, ammonium or antibiotic action. The use of sensors allowed to continuously record, in growing cultures, even small changes in parameters. There were significant differences in the amount and kinetics of sulfide production between Bacillus and E. coli. These differences are thought to be due to the lack of glutathione in Bacillus. It is suggested that stress-induced sulfide production by Bacillus under the described conditions may be one of the previously unknown sources of hydrogen sulfide in nature. [ABSTRACT FROM AUTHOR]

Carajasia cangae (Rubiaceae) is a narrow endemic species from the canga ecosystems of the Carajás National Forest that is facing extinction due to a limited range and habitat disturbance from hydroclimatological changes and mining activities. This study examines the influence of rhizospheric and endophytic bacteria on C. cangae seed germination to support conservation efforts. Soil samples, both rhizospheric and non-rhizospheric, as well as plant root tissues, were collected. Bacteria from these samples were subsequently isolated, cultured, and identified. DNA sequencing revealed the presence of 16 isolates (9 rhizospheric and 7 endophytic), representing 19 genera and 6 phyla: Proteobacteria, Actinobacteria, Acidobacteria, Firmicutes, Bacteroidetes, and Chloroflexi. The endophytic isolates of Bacillus and the rhizospheric isolates of Planococcus and Lysinibacillus reduced the median germination time and initiation time, while the rhizospheric isolates Serratia and Comamonas increased the germination time and decreased the germination percentage in comparison to the control sample. These findings emphasize the crucial role of endophytic bacteria in the germination of C. cangae and highlight isolates that could have beneficial effects in the following stages of plant growth. Understanding the impact of endophytic and rhizospheric bacterial isolates on seed germination can enhance conservation efforts by shortening the germination period of this species and thereby improving seedling production. Additionally, this knowledge will pave the way for future research on the role of bacteria in the establishment of C. cangae. [ABSTRACT FROM AUTHOR]

Salvinia auriculata Aublet is a floating aquatic plant, capable of absorbing the excess of nutrients and water contaminants and can be used in effluent treatment plants. The ability to survive in degraded areas may be related to the association with beneficial bacteria capable of promoting plant growth. However, little is known about the microbiota associated with this aquatic plant and its potential application to the aquatic environment. In this sense, this work aims to identify bacteria associated with S. auriculata that could be able to promote plant growth. Eighteen bacterial strains were identified by sequencing of the 16S rRNA gene, belonging to the genera Agrobacterium, Bacillus, Curtobacterium, Enterobacter, Pseudomonas, Siccibacter, and Stenotrophomonas. All isolates produced indole compounds, 12 fixed N2, and 16 solubilized phosphate. A new strain of Enterobacter (sp 3.1.3.0.X.18) was selected for inoculation into S. auriculata. For this purpose, 500 mL of nutrient solution and 1 g of the plant were used in the control and inoculated conditions. Enterobacter inoculation promoted a significant increase (p ≤ 0.05) in fresh plant biomass (17%) after 4 days of cultivation. In summary, the present study characterized 18 plant-growth-promoting bacteria isolated from S. auriculata with potential for biotechnological application, such as the production of bioinoculants or biomass resources, to protect or improve plant growth under conditions of stress. [ABSTRACT FROM AUTHOR]

Biofertilizers are environmentally friendly compounds that can enhance plant growth and substitute for chemically synthesized products. In this research, a new strain of the bacterium Bacillus velezensis, designated JZ, was isolated from the roots of strawberry plants and exhibited potent antagonistic properties against Bacillus altitudinis m-1, a pathogen responsible for leaf spot disease in strawberry. The fermentation broth of JZ exerted an inhibition rate of 47.43% against this pathogen. Using an optimized acid precipitation method, crude extracts of lipopeptides from the JZ fermentation broth were obtained. The crude extract of B. velezensis JZ fermentation broth did not significantly disrupt the cell permeability of B. altitudinis m-1, whereas it notably reduced the Ca2+-ATPase activity on the cell membrane and markedly elevated the intracellular reactive oxygen species (ROS) concentration. To identify the active compounds within the crude extract, QTOF-MS/MS was employed, revealing four antimicrobial compounds: fengycin, iturin, surfactin, and a polyene antibiotic known as bacillaene. The strain JZ also produced various plant-growth-promoting substances, such as protease, IAA, and siderophore, which assists plants to survive under pathogen infection. These findings suggest that the JZ strain holds significant potential as a biological control agent against B. altitudinis, providing a promising avenue for the management of plant bacterial disease. [ABSTRACT FROM AUTHOR]

A novel aerobic denitrifying bacterium Bacillus cereus J1 was isolated from a sewage treatment plant. Its characteristics under two distinct nitrogen sources were systematically investigated. According to the results of whole-genome sequencing, we inferred that strain J1 removes nitrogen through processes such as aerobic denitrification, dissimilatory nitrate reduction to ammonium, and ammonium assimilation. The degradation process of COD and total inorganic nitrogen (TIN) correlated to the zero-order degradation kinetics equation, and the maximum removal rate of NO3−−N reached 3.17 mg/L/h and that of NH4+−N was 3.79 mg/L/h. Utilizing single-factor experiments and response surface methodology, the optimal conditions for nitrate removal were determined as a shaking speed of 115 rpm, COD/nitrogen mass (C/N ratio) of 12.25, and salinity of 3.44 g/L, with the C/N ratio exerting the most significant influence. Similarly, for the maximum ammonium removal, the ideal conditions involved a shaking speed of 133 rpm, C/N ratio of 29, and salinity of 13.30 g/L, with the shaking speed exerting the most significant influence. These findings demonstrate that large amounts of ammonium and nitrate can be quickly removed with the help of Bacillus cereus J1, indicating that strain J1 may be applied to alleviate nitrogen pollution in aquatic environments. [ABSTRACT FROM AUTHOR]

Synthetic microbial communities (SynComs) have been shown to be an ecofriendly alternative for promoting plant growth. However, the mechanisms by which SynCom inoculants drive plant growth promotion in rhizosphere soil are still not fully explored. Herein, we designed a three-strain consortium based on the biocompatibility among strains and indole-3-acetic acid (IAA) production. The consortium containing Bacillus safensis 5-49, Bacillus stratosphericus 5-54, and Bacillus halotolerans 6-30 possessed a synergistic effect on IAA production and biofilm formation. Genetic analysis suggested that IAA was synthesized through tryptophan-dependent pathways in the strains. The consortium outperformed the plant growth-promoting effect observed with single strains, showing an increase in walnut (Juglans regia) seedling dry weight by 92.3% over the non-inoculated plants after 60 days of cultivation. This effect was underpinned by the synergistic interactions of the consortium, which was evidenced by the significantly increased relative abundance of Bacillus and tryptophan metabolism-associated genes in the rhizosphere of consortium-inoculated plants. Meanwhile, the consortium increased the relative abundance of indigenous Pseudomonas in rhizosphere soil, providing a synergistic effect on improving soil enzyme activities and thus available nutrients. The available N, P, and K contents in the consortium-inoculated plant rhizosphere were 3.77–28.4% higher than those in non-inoculated samples. This work provided an efficient bacterial consortium and proposed the mode of action by which this consortium improved plant growth and soil fertility. [ABSTRACT FROM AUTHOR]

Maize, one of the most important cereal crops in Bangladesh, is severely contaminated by fumonisin, a carcinogenic secondary metabolite produced by Fusarium including Fusarium proliferatum. Biocontrol with Bacillus strains is an effective approach to controlling this F. proliferatum as Bacillus has proven antagonistic properties against this fungus. Therefore, the present study aimed to determine how native Bacillus strains can reduce fumonisin in maize cultivated in Bangladesh, where BDISO76MR (Bacillus subtilis) strains showed the highest efficacy both in vitro in detached cob and in planta under field conditions. The BDISO76MR strain could reduce the fumonisin concentration in detached cob at 98.52% over untreated control, by inhibiting the conidia germination and spore formation of F. proliferatum at 61.56% and 77.01%, respectively in vitro. On the other hand, seed treatment with formulated BDISO76MR showed higher efficacy with a reduction of 97.27% fumonisin contamination compared to the in planta cob inoculation (95.45%) over untreated control. This implies that Bacillus-based formulation might be a potential approach in mitigating fumonisin contamination in maize to ensure safe food and feed. [ABSTRACT FROM AUTHOR]