Search Results (533)

Common bean (Phaseolus vulgaris L.) is a critical protein-rich legume supporting food and nutritional security globally. However, Fusarium wilt, caused by Fusarium solani, remains a major constraint to production, with yield losses reaching up to 84%. While biocontrol strategies have been explored, most microbial agents are sourced from mesophilic environments and show limited effectiveness under abiotic stress. Here, we report the isolation and characterization of extremophilic Bacillus spp. from the hypersaline Lake Bogoria, Kenya, and their biocontrol potential against F. solani. From 30 isolates obtained via serial dilution, 9 exhibited antagonistic activity in vitro, with mycelial inhibition ranging from 1.07–1.93 cm 16S rRNA sequencing revealed taxonomic diversity within the Bacillus genus, including unique extremotolerant strains. Molecular screening identified genes associated with the biosynthesis of antifungal metabolites such as 2,4-diacetylphloroglucinol, pyrrolnitrin, and hydrogen cyanide. Enzyme assays confirmed substantial production of chitinase (1.33–3160 U/mL) and chitosanase (10.62–28.33 mm), supporting a cell wall-targeted antagonism mechanism. In planta assays with the lead isolate (B7) significantly reduced disease incidence (8–35%) and wilt severity (1–5 affected plants), while enhancing root colonization under pathogen pressure. These findings demonstrate that extremophile-derived Bacillus spp. possess robust antifungal traits and highlight their potential as climate-resilient biocontrol agents for sustainable bean production in arid and semi-arid agroecosystems. Full article

A simple and rapid precipitation process was successfully employed to prepare silver phosphate (SP, Ag₃PO₄). Two different phosphate sources: diammonium hydrogen phosphate ((NH₄)₂HPO₄) and dipotassium hydrogen phosphate (K₂HPO₄) were applied separately as the precursor, obtaining ((NH₄)₂HPO₄)⁻ and K₂HPO₄⁻ derived SP powders, named SP-A or SP-P, respectively. Fourier transform infrared (FTIR) spectra pointed out the vibrational characteristics of P–O and O–P–O interactions, confirming the presence of the PO43– functional group for SP. X-ray diffraction (XRD) patterns revealed that the SP crystallized in a cubic crystal structure. Whereas the field emission scanning electron microscope (FESEM) exposed spherical SP particles. The potentially antibacterial activity of SP-A and SP-P against bacterial Bacillus stratosphericus, yeast Meyerozyma guilliermondii, and fungal Phanerodontia chrysosporium was subsequently investigated. All studied microorganisms were recovered and isolated from the aquatic plant during the tissue culture process. The preliminary result of the antimicrobial test revealed that SP-A has higher antimicrobial activity than SP-P. The superior antimicrobial efficiency of SP-A compared to SP-P may be attributed to its purity and crystallite size, which provide a higher surface area and more active sites. In addition, the presence of potassium-related impurities in SP-P could have negatively affected its antimicrobial performance. These findings suggest that SP holds potential as an antimicrobial agent for maintaining sterility in tissue cultures, particularly in aquatic plant systems. The growth of both B. stratosphericus and M. guilliermondii was suppressed effectively at 30 ppm SP-A, whereas 10 ppm of SP-A can suppress P. chrysosporium development. This present work also highlights the potential of SP at very low concentrations (10–30 ppm) for utilization as an effective antimicrobial agent in tissue culture, compared to a commercial antimicrobial agent, viz., acetic acid, at the same concentration. Full article

Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential against L. monocytogenes. Methods: A total of 26 Bacillus isolates were screened for antimicrobial activity, gastrointestinal resilience, and host cell adhesion. A fluorescence-based infection assay using mCherry-expressing HCT 116 cells was used to assess cytoprotection against L. monocytogenes NCTC 7973. Eight strains significantly improved host cell viability and were validated by quantification of intracellular CFU. Two top candidates were tested in a murine model of listeriosis. The genome of the lead strain was sequenced to evaluate safety and biosynthetic potential. Results: B. subtilis CECT 8266 completely inhibited intracellular replication of L. monocytogenes in HCT 116 cells, reducing bacterial recovery to undetectable levels. In vivo, it decreased splenic bacterial burden by approximately 6-fold. Genomic analysis revealed eight bacteriocin biosynthetic clusters and silent antibiotic resistance genes within predicted genomic islands, as determined by CARD and Alien Hunter analysis. The strain also demonstrated bile and acid tolerance, as well as strong adhesion to epithelial cells. Conclusions: The proposed pipeline enables efficient identification of probiotic Bacillus strains with intracellular protective activity. B. subtilis CECT 8266 is a promising candidate for translational applications in food safety or health due to its efficacy, resilience, and safety profile. Full article

Hydrazine, a highly toxic and reactive compound widely used as rocket fuel, poses significant environmental and health risks, particularly in long-term space missions. This study investigates the cometabolic capacity of Chlorella vulgaris and seven extremophilic Bacillus spp. strains—isolated from the arid Dead Sea region—to tolerate and degrade hydrazine at concentrations up to 25 ppm. The microalga C. vulgaris reduced hydrazine levels by 81% within 24 h at 20 ppm, while the Bacillus isolates achieved an average reduction of 45% over 120 h. Identified strains included B. licheniformis, B. cereus, and B. atrophaeus. Co-culture experiments demonstrated that C. vulgaris and B. cereus (isolate ISO-36) stably coexisted without antagonistic effects, suggesting a synergistic detoxification interaction. Flow cytometry revealed that most bacteria transitioned into spores under stress, highlighting a survival adaptation. Titanium, representing a biocompatible material common in aerospace hardware, did not inhibit microbial growth or hydrazine degradation. These findings underscore the potential of Dead Sea-derived microbial consortia for cometabolic hydrazine detoxification and support the feasibility of converting spacecraft components into functional photobioreactors. This approach offers dual-use benefits for space missions and industrial wastewater treatment. Future studies should investigate degradation pathways, stress resilience, and bioreactor scale-up. Full article

Following hygiene standards in milk production is essential for making high-quality sour cream, especially when using traditional methods that rely on raw milk. The aim of this study was to determine the physicochemical, microbiological, and sensory quality of artisanal sour cream samples collected from major marketplaces in the wider Zagreb area. On average, the samples contained 27.99% milk fat, 3.30% protein, 34.29% dry matter, 6.51% fat-free dry matter and 3.00% lactose, with considerable variability observed across all components. Microbiological analysis revealed the presence of Staphylococcus aureus in 35.30% of the samples, Enterobacteriaceae in 76.47%, Escherichia coli in 94.11%, Bacillus spp. in 23.53%, and yeasts in 100% of the samples. Listeria monocytogenes and Salmonella spp. were not detected. The sensory analysis of the textural properties showed significant variability in firmness, adhesiveness, viscosity, creaminess, and fizziness. Samples with higher milk fat and dry matter content were rated better for creaminess, viscosity and mouth firmness. Flavour assessments, particularly for cream and diacetyl notes, also varied widely among samples. These findings highlight the complexity of sour cream’s sensory attributes and the significant influence of ingredient composition and processing techniques on appearance, aroma, texture, taste, and flavour. Principal component analysis (PCA) with Varimax rotation simplified the data structure and identified key dimensions of quality variation. Principal component analysis (PCA) revealed that the first principal component (PC1) effectively discriminated the cream samples based on sensory attractiveness and indicators of spoilage and highlighted the association between off-flavour and microbial contamination with inferior characteristics. The second principal component (PC2) captured the differences in physicochemical characteristics and showed a gradient from richer, creamier samples with higher fat content to those with lower acidity and higher freshness. Full article

The spoilage of dairy products and foodborne diseases caused by Bacillus spp. are important public concerns. The objective of this study was to estimate the global prevalence of Bacillus spp. in a range of milk and dairy products by using a meta-analysis of literature data published between 2001 and 2023. A total of 3624 publications were collected from Web of Science and PubMed databases. Following the principles of systematic review, 417 sets of prevalence data were extracted from 142 eligible publications. Estimated by the random-effects model, the overall prevalence of Bacillus spp. in milk and dairy products was 11.8% (95% CI: 10.1–13.7%), with highly severe heterogeneity (94.8%). Subgroup analyses revealed substantial heterogeneity in Bacillus spp. prevalence according to geographical continents, sources of sampling, types of dairy products, microbial species, and detection methods. The prevalence of Bacillus spp. was highest in Asia (15.4%, 95% CI: 12.3–19.1%), lowest in Oceania (3.5%, 95% CI: 3.3–3.7%) and generally higher in developing versus developed countries. The prevalence of Bacillus spp. isolated from retail markets (16.1%, 95% CI: 13.0–19.7%) was higher than from farms (10.3%, 95% CI: 6.9–15.0%) or dairy plants (9.2%, 95% CI: 7.1–12.0%). This finding is likely attributable to its inherent characteristic of the resistant endospores and ubiquitous presence in the environment—Bacillus spp. can potentially cyclically contaminate farms, dairy products and human markets. Regarding the species distribution, Bacillus cereus presented a cosmopolitan distribution across all continents. The epidemic patterns of different Bacillus species vary depending on the sample sources. In addition, the detection method utilized also affected the reported prevalence of Bacillus spp. It is recommended to use molecular-based rapid detection methods to obtain a more accurate prevalence of Bacillus contamination. Therefore, a better understanding of variations in Bacillus spp. prevalence across different factors will enable competent authorities, industries, and other relevant stakeholders to tailor their interventions for effectively controlling Bacillus spp. in milk and dairy products. Full article

The cultivation of cowpea (Vigna unguiculata), a vital vegetable crop, faces significant threats from Fusarium spp.-induced root rot. In this study, three fungal pathogens (Fusarium falciforme HKFf, Fusarium incarnatum HKFi, and Fusarium oxysporum HKFo) were isolated from symptomatic cowpea plants, and we screened 90 rhizobacteria from healthy rhizospheres using six culture media. Among these pathogens, Priestia megaterium TSA-10E showed a notable suppression of F. oxysporum HKFo (63.21%), F. incarnatum HKFi (55.16%), and F. falciforme HKFf (50.93%). In addition, Bacillus cereus KB-6 inhibited the mycelial growth of F. incarnatum HKFi and F. oxysporum HKFo by 42.39% and 47.93%, respectively. Critically, cell-free filtrates from P. megaterium TSA-10E and B. cereus KB-6 cultures reduced conidial germination in F. oxysporum HKFo and F. incarnatum HKFi, highlighting their role in disrupting the early infection stages. In greenhouse trials, TSA-10E and KB-6 reduced disease severity by 48.7% and 40.4%, respectively, with treated plants maintaining healthy growth while untreated controls succumbed to wilting. Broad-spectrum assays revealed that B. subtilis TSA-6E and P. megaterium TSA-10E were potent antagonists against both economic and grain crop pathogens. These findings underscore the potential of rhizobacteria as sustainable biocontrol agents for managing root rot disease caused by Fusarium spp. in cowpea cultivation. Full article

Surfactants are widely utilized in agriculture as emulsifying, dispersing, anti-foaming, and wetting agents. In these adjuvant roles, the inherent biological activity of the surfactant is secondary to the active ingredients. Here, the hydrophilic non-ionic surface-active tri-block copolymer Pluronic^® F68 is investigated for direct biological activity in wheat. F68 binds to and inserts into lipid membranes, which may benefit crops under abiotic stress. F68’s interactions with Triticum aestivum (var Juniper) seedlings and a seed-borne Bacillus spp. endophyte are presented. At concentrations below 10 g/L, F68-primed wheat seeds exhibited unchanged emergence. Root-applied fluorescein-F68 (fF68) was internalized in root epidermal cells and concentrated in highly mobile endosomes. The potential benefit of F68 in droughted wheat was examined and contrasted with wheat treated with the osmolyte, glycine betaine (GB). Photosystem II activity of droughted plants dropped significantly below non-droughted controls, and no clear benefit of F68 (or GB) during drought or rehydration was observed. However, F68-treated wheat exhibited increased transpiration values (for watered plants only) and enhanced shoot dry mass (for watered and droughted plants), not observed for GB-treated or untreated plants. The release of seed-borne bacterial endophytes into the spermosphere of germinating seeds was not affected by F68 (for F68-primed seeds as well as F68 applied to roots), and the planktonic growth of a purified Bacillus spp. seed endophyte was not reduced by F68 applied below the critical micelle concentration. These studies demonstrated that F68 entered wheat root cells, concentrated in endosomes involved in transport, significantly promoted shoot growth, and showed no adverse effects to plant-associated bacteria. Full article

The excessive use of chemical fertilizers in tea cultivation threatens soil health, environmental sustainability, and long-term crop productivity. This study explores the application of plant growth-promoting bacteria (PGPB) as an eco-friendly alternative to conventional fertilizers. A bacterial consortium was developed using selected rhizobacterial isolates—Lysinibacillus fusiformis, five strains of Serratia marcescens, and two Bacillus spp.—based on their phosphate and zinc solubilization abilities and production of ACC deaminase, indole-3-acetic acid, and siderophores. The consortium was tested in both pot and field conditions using two tea clones, S3A3 and TS491, and compared with a chemical fertilizer treatment. Plants treated with the consortium showed enhanced growth, biomass, and antioxidant activity. The total phenolic contents increased to 1643.6 mg GAE/mL (S3A3) and 1646.93 mg GAE/mL (TS491), with higher catalase (458.17–458.74 U/g/min), glutathione (34.67–42.67 µmol/gfw), and superoxide dismutase (679.85–552.28 units/gfw/s) activities. A soil metagenomic analysis revealed increased microbial diversity and the enrichment of phyla, including Acidobacteria, Proteobacteria, Actinobacteria, Chloroflexi, and Firmicutes. Functional gene analysis showed the increased abundance of genes for siderophore biosynthesis, glutathione and nitrogen metabolism, and indole alkaloid biosynthesis. This study recommends the potential of a PGPB consortium as a sustainable alternative to chemical fertilizers, enhancing both the tea plant performance and soil microbial health. Full article

Coinfection poses severe threats to poultry health, particularly due to the complexity and resilience of multispecies interactions, increasing the difficulty of treatment. Haemophilus spp., a heterotrophic bacterium, heavily relies on extracellular growth factors acquired from other organisms or its surrounding environment. Although coinfections by Avibacterium paragallinarum and Bacillus have been reported, the underlying mechanism of the cooperative interaction remains poorly understood. In this study, we characterized the growth-promoting properties and nicotinamide adenine dinucleotide production of some Bacillus species, including probiotic Bacillus, to evaluate the feasibility of A. paragallinarum coinfection in vitro. Meanwhile, we determined the minimum inhibitory concentration (MIC) and population dynamics of cocultured Bacillus and A. paragallinarum to assess the effect of bacterial interactions on antibiotic efficacy. Additionally, we demonstrated that B. cereus aggravates rhinitis symptoms in chickens infected with A. paragallinarum. Our findings reveal that Bacillus spp.-derived metabolites sustain A. paragallinarum growth and enhance its survival, thereby highlighting the infection risks associated with Bacillus colonization in the respiratory tract. Full article

Plant microbiota contributes to nutrient absorption, and the production of hormones and vitamins, and plays a crucial role in responding to environmental stress. We hypothesized that Vaccinium spp. harbour a unique microbiota that enables them to coexist in extreme environments such as saline, nutrient-poor, and waterlogged conditions. Upon examining Bacillus spp. endophytes isolated from blueberries, cranberries and lingonberries in vitro, we identified B. halotolerans (Bil-LT1_1, Bil-LT1_2) and B. velezensis (Cran-LT1_8, Ling-NOR4_15) strains that inhibit the growth of five pathogenic fungi and five foodborne bacteria. Whole-genome sequencing provided insights into genome organization and plasticity, helping identify mobile elements and genes potentially acquired through horizontal gene transfer. Functional annotation identified genes associated with plant colonization, stress tolerance, biocontrol activity, and plant growth promotion. Comparative genomic analyses revealed key biosynthetic gene clusters (BGCs) responsible for producing antifungal metabolites, including lipopeptides and polyketides. Genes supporting plant nutrition, growth, and environmental adaptation were present also in these strains. Notably, isolated endophytes exhibited particularly high levels of genomic plasticity, likely due to horizontal gene transfer involving gene ontology (GO) pathways related to survival in polymicrobial and foreign environments. Full article

This study assessed the viable and culturable microbial diversity that remained on equipment surfaces after hygiene procedures in Brazilian poultry and fish slaughterhouses. Food-contact surface samples were collected using sterile swabs in poultry (n = 50) and fish (Oreochromis niloticus, n = 50) slaughterhouses. The swab samples were used to prepare culture plates to recover viable and culturable cells. The grown plates were washed, and the total DNA of the cell suspension was extracted with a commercial kit. Sequencing of the total DNA extracted from cultures was targeted at the V3 and V4 regions of the 16S rRNA. DNA reads were analyzed by QIIME2 software, with results expressed in relative frequency (%RF). Alpha and beta diversity indexes were analyzed considering the spots of sample collection, type of industry, surfaces (smooth or modular), and materials (polypropylene, stainless steel, or polyurethane). The results showed that in the poultry slaughterhouse, the most abundant genera were Acinetobacter (27.4%), Staphylococcus (7.7%), and Pseudomonas (5.3%), while for the fish slaughterhouse, there was a higher abundance of Staphylococcus (27.7%), Acinetobacter (17.2%), and Bacillus (12.5%). Surface characteristics influenced the microbial diversity, with Acinetobacter spp. dominating modular surfaces and Staphylococcus spp. prevailing on smooth surfaces. The results obtained indicate there is an important resident microbiota that persists even after hygiene processes, and surface-specific cleaning strategies should be developed. Full article

The major objective was to investigate the protective capabilities of endophytic bacterial strains isolated from a number of medicinal plant species towards Aspergillus spp. secured from the internal tissues of fungi-infected peanuts. Among 32 fungal isolates surveyed for mycotoxin production in various culture media (PDA, RBCA, YES, CA), 10 isolates qualitatively producing AFB₁, besides 10 OTA-producers, were assayed by HPLC for quantitative toxin production. Aspergillus spp. isolate Be 13 produced an extraordinary quantity of 1859.18 μg mL⁻¹ AFB₁, against the lowest toxin level of 280.40 μg mL⁻¹ produced by the fungus isolate IS 4. The estimated amounts of OTA were considerably lower and fell in the range 0.88–6.00 μg mL⁻¹; isolate Sa 1 was superior, while isolate Be 7 seemed inferior. Based on ITS gene sequencing, the highly toxigenic Aspergillus spp. isolates Be 13 and Sa 1 matched the description of A. novoparasiticus and A. ochraceus, respectively, ochraceus, respectively, which are present in GenBank with identity exceeding 99%. According to 16S rRNA gene sequencing, these antagonists labeled Ar6, Ma27 and So34 showed the typical characteristics of Pseudomonas aeruginosa, Bacillus subtilis and Bacillus velezensis, respectively, with similarity percentages of 99–100. The plant growth-promoting activity measurements of the identified endophytes indicated the production of 16.96–80.00 μg/100 mL culture medium of IAA. Phosphate-solubilizing capacity varied among endophytes from 2.50 to 21.38 μg/100 mL. The polysaccharide production pool of bacterial strains ranged between 2.74 and 6.57 mg mL⁻¹. P. aeruginosa Ar6 and B. velezensis successfully produced HCN, but B. subtilis failed. The in vitro mycotoxin biodegradation potential of tested bacterial endophytes indicated the superiority of B. velezensis in degrading both mycotoxins (AFB₁-OTA) with average percentage of 88.7; B. subtilis ranked thereafter (85.6%). The 30-day old peanut (cv. Giza 6) seedlings grown in gnotobiotic system severely injured due to infection with AFB₁/OTA-producing fungi, an effect expressed in significant reductions in shoot and root growth traits. Simultaneous treatment with the endophytic antagonists greatly diminished the harmful impact of the pathogens; B. velezensis was the pioneer, not P. aeruginosa Ar6. In conclusion, these findings proved that several endophytic bacterial species have the potential as alternative tools to chemical fungicides for protecting agricultural commodities against mycotoxin-producing fungi. Full article

This study aims to identify the optimal conditions—host, plant material, seasonality, and agricultural practices—for isolating and developing a collection of culturable endophytic microorganisms to support sustainable Olea europaea L. cultivation. Samples were collected from three Sicilian olive cultivars (‘Nocellara del Belice’, ‘Nocellara Etnea’, and ‘Nocellara Messinese’) and six wild olive accessions across different phenological phases and under organic and conventional agronomic management. Endophytes were isolated from leaves and twigs using a culture-dependent approach, and their taxonomic diversity and plant-growth-promoting (PGP) traits were analyzed. A total of 133 endophytic isolates were identified, spanning bacterial (Proteobacteria, Firmicutes, and Actinobacteria) and fungal (Ascomycota and Basidiomycota) phyla. Wild olive trees contributed more than cultivated varieties to enriching the diversity and composition of culturable endophyte collection as well as twigs instead of leaves. Winter sampling allowed to implement the taxonomic genera of olive endophyte collection. Both farming systems favored an increase in the composition of microbial collection, though organic farming systems supported greater microbial richness. Functional analysis highlighted key PGP traits in a selection of bacterial isolates, including indole-3-acetic acid and siderophore production, nitrogen fixation, and antifungal activity. Bacillus spp. dominated enzymatic activities, such as amylase, protease, and lipase production, as well as antifungal activity against the olive fungal pathogen Neofusicoccum vitifusiforme. This research highlights the significant diversity and functional potential of Mediterranean olive endophytes. Our findings emphasize the role of native microbial communities as bio-inoculants, promoting plant growth, nutrient uptake, and disease resistance. These insights lay the groundwork for developing targeted olive-microbial consortia for biocontrol and stress tolerance applications. Full article

Norway spruce (Picea abies (L.) Karst.) has been widely planted beyond its natural range due to its fast growth rate and valuable wood. In Québec, over 200 million seedlings have been planted since 1964. Several of these plantations are now facing a new potential threat, i.e., spruce budworm (Choristoneura fumiferana (Clem.)) infestations. Despite contrasting results, Norway and white spruce (P. glauca [Moench] Voss) apparently sustain a similar degree of budworm defoliation. The main study objective is to quantify defoliation in Norway spruce caused by spruce budworm. We also evaluate the efficacy of Bacillus thuringiensis Berliner spp. kurstaki (Btk) in protecting this exotic host tree. Annual defoliation was assessed in plantations of Norway, white, and black spruce (P. mariana [Mill.] BSP) between 2018 and 2022 in the Bas-Saint-Laurent region. Additional surveys were conducted in Norway and white spruce plantations in the Gaspésie and Côte-Nord to evaluate Btk efficacy. We show that both species exhibit similar defoliation levels, though Norway spruce sometimes sustains greater damage (e.g., 35% vs. 10% in 2019). Btk formulations showed low efficacy in protecting Norway spruce foliage (≥49.32% defoliation in treated plantations). Further studies are needed to understand factors influencing Btk efficacy on this host. Full article