Search Results (28)

Soil salinization represents a significant abiotic constraint to global agricultural sustainability. The potential of extremophile plant growth-promoting bacteria (PGPB) to alleviate such stress in maize was investigated in this study. Siderophore-producing PGPB enhance plant growth and improve the rhizosphere microenvironment by increasing nutrient availability and inducing systemic resistance. Two salt-tolerant, high-siderophore-producing PGPB strains, Bacillus toyonensis TRM58010 and Peribacillus frigoritolerans TRM58009, were isolated and identified from soil samples collected on the Pamir Plateau. In this study, we found that B. toyonensis TRM58010 synthesized catechol-type siderophores, which enhanced iron availability for maize in saline–alkaline conditions, thereby improving iron nutrition and directly promoting root and stem growth under salt stress. P. frigoritolerans TRM58009 produced hydroxamate-type siderophores, which increased maize iron uptake and stimulated antioxidant enzyme activity, mitigating oxidative stress caused by salinity and alkalinity and supporting overall plant health. Both strains demonstrated robust tolerance to extreme alkaline and saline conditions. Hydroponic and pot experiments showed that these strains significantly improved maize germination rate, root and stem development, plant height, leaf growth, antioxidant enzyme activities, and chlorophyll content under saline–alkaline stress. Notably, the application of P. frigoritolerans TRM58009 bacterial suspension increased maize leaf catalase, peroxidase, and superoxide dismutase activities by 15.712%, 11.584%, and 2.820%, respectively (all p < 0.05), while decreasing malondialdehyde (MDA) content by 15.685% (p < 0.05). P. frigoritolerans TRM58009 elevated chlorophyll a content by 23.4% (p < 0.05). These findings demonstrate the potential of extremophile PGPB strains to mitigate the impact of saline–alkaline stress on maize growth. The distinct growth-promoting effects of these strains, isolated from Pamir Plateau meadow soils, present a promising strategy for bioremediation of saline–alkaline lands and the development of efficient microbial fertilizers. By advancing the use of salt-tolerant siderophore-producing bacteria, this study lays the foundation for innovative approaches to enhance crop resilience and productivity in challenging environments. Full article

The Bacillus cereus group (B. cereus group) comprises several closely related species that share high genetic similarity but display markedly different phenotypic traits and pathogenic potential. Reliable and rapid discrimination at the species level remains challenging using conventional microbiological and molecular methods. In this study, Fourier Transform Infrared (FTIR) spectroscopy was evaluated as a rapid phenotypic approach to differentiate seven members of the Bacillus cereus sensu stricto (B. cereus s.s.), Bacillus anthracis (B. anthracis), Bacillus thuringiensis (B. thuringensis), Bacillus mycoides (B. mycoides), Bacillus toyonensis (B. toyonensis), Bacillus wiedmannii (B. wiedmannii) and Bacillus weihenstephanensis (B. weihenstephanensis). A collection of 190 isolates previously characterized by whole genome sequencing was analyzed using the IR Biotyper system. Spectral data were processed through multivariate analyses, including principal component analysis and linear discriminant analysis, following a hierarchical classification strategy. FTIR spectroscopy enabled clear discrimination of B. anthracis from other members of the B. cereus group and allowed the separation of several additional species based on distinct spectral signatures. A further discrimination step permitted differentiation between B. cereus sensu stricto and B. thuringiensis, with minimal overlap. These findings demonstrate that FTIR spectroscopy represents a promising and rapid tool for species-level discrimination within the B. cereus group. While the results should be considered preliminary for species represented by a limited number of isolates, this approach shows strong potential as a complementary method to molecular techniques in routine diagnostics in food safety and veterinary microbiology. Full article

As an important complementation of plant genetic traits, seed endophytes (SEs) have garnered significant attention due to their crucial roles in plant germination and early seedling establishment. In this study, we employed both culture-dependent and amplicon sequencing-based approaches to characterize the endophytic microbiome in seed samples derived from different individual Panax notoginseng plants. Additionally, we evaluated the antagonistic activity of isolated culturable bacterial SEs against the root rot pathogens Fusarium solani and F. oxysporum. Our results demonstrated that a greater sampling quantity substantially increased the species richness (Observed OTUs) and diversity of seed endophytic microbiota, underscoring the importance of seed population size in facilitating the vertical transmission of diverse endophytes to progeny. The endophytic communities (including both fungi and bacteria) exhibited a conserved core microbiota alongside host-specific rare taxa, forming a phylogenetically and functionally diverse endophytic resource pool. Core bacterial genera included Streptococcus, Methylobacterium-Methylorubrum, Sphingomonas, Burkholderia-Caballeronia-Paraburkholderia, Pantoea, Halomonas, Acinetobacter, Pseudomonas, Vibrio, and Luteibacter, while core fungal genera comprised Davidiella, Thermomyces, Botryotinia, Myrothecium, Haematonectria, and Chaetomium. Among 256 isolated endophytic bacterial strains, 11 exhibited strong inhibitory effects on the mycelial growth of F. solani and F. oxysporum. Further evaluation revealed that two antagonistic strains, Bacillus cereus and B. toyonensis, significantly enhanced seed germination and plant growth in P. notoginseng, and effectively suppressed root rot disease in seedlings. These findings highlight the potential use of SEs as biocontrol agents and growth promoters in sustainable agriculture. Full article

In vitro shoot culture and cryopreservation (CP) are techniques essential for the ex situ preservation of genetic resources and the production of plant propagation material of clonally propagated horticultural crops. Changes in plant-associated microbiota diversity and composition induced by in vitro cultivation and CP treatment could have a negative effect on the growth and ex vitro adaptation of the in vitro propagated shoots. Therefore, the aim of the present study was to assess changes in endophytic bacteria diversity in domestic apple tissues induced by in vitro cultivation and CP treatment and to investigate the potential of the bacterial inoculum to improve the rooting and ex vitro acclimatisation of the propagated shoots. Metataxonomic analysis revealed a variation in the endophytic bacteria diversity and taxonomic composition between the field-grown tree dormant bud and the in vitro propagated or CP-treated shoot samples of apple cv. Gala. Whereas Sphingobacteriaceae, Sphingomonadaceae, Pseudomonadaceae, and Beijerinckiaceae families were the most prevalent families in the bud samples, Enterobacteriaceae, Bacillaceae, and Lactobacillaceae were dominant in the in vitro shoots. The bacterial inoculum effect on rooting and ex vitro acclimatisation was assessed using four isolates selected by screening the endophytic isolate collection. Bacillus sp. L3.4, B. toyonensis Nt18, or a combined inoculum resulted in a 21%, 36%, and 59% increase in cumulative root length and a 41%, 46%, and 35% increase in the biomass accumulation of ex vitro acclimatised plantlets, respectively. Root zone microbiota functional diversity analysis implied that growth stimulation was not related to improved nutrient uptake but could involve a pathogen-suppressing effect. The results demonstrate that the application of plant growth-promoting bacteria can potentially improve the performance of the in vitro propagated germplasm. Full article

This study evaluated the effects of Bacillus toyonensis BCT-7112^T on laying duck performance, egg quality, ammonia emission, and cecal microbiota. Two hundred twenty 32-week-old Khaki Campbell ducks were assigned to four dietary treatments (0, 200, 500, or 1000 ppm Toyocerin^® 10⁹ premixture, containing 1 × 10⁹ B. toyonensis BCT-7112^T CFU/g) with five replicate pens of eleven birds each. Data on productivity, egg quality, and ammonia emissions were collected weekly, and cecal microbiota were analyzed at 44 weeks of age. Supplementation with 1000 ppm B. toyonensis BCT-7112^T significantly increased average egg weight (p < 0.001) and eggshell thickness (p = 0.007). Egg mass also improved at higher supplementation levels (p < 0.05), though feed intake, feed conversion, daily egg production, and most egg quality parameters were unaffected (p > 0.05). Ammonia emissions in litter decreased significantly (p < 0.05) with probiotic inclusion. Cecal microbiota analysis revealed higher diversity in ducks fed 1000 ppm, with a notable shift in predominant phyla from Bacteroidetes (35.12%) and Firmicutes (34.93%) in the controls to Bacteroidetes (40.52%), Firmicutes (34.08%), and Deferribacteres (9.54%) in the treated ducks. The findings suggest that 1000 ppm B. toyonensis BCT-7112^T enhances egg production, eggshell quality, and microbial diversity while reducing ammonia emissions in laying duck systems. Full article

Bacillus cereus sensu lato can contaminate food and cause food poisoning by producing toxins such as cereulide, toxin BL, and cytotoxin K. In this study, we retrospectively analyzed B. cereus sensu lato from retail food products and food poisoning cases using PCR methods to determine their virulence profiles. A new toxin profile, encoding all four toxins (hbl, nhe, cytK, ces), was found in 0.4% of isolates. The toxin profiles, classified into A-J, revealed that 91.8% harbored nhe genes, while hbl, cytK, and ces were detected in 43.8%, 46.9%, and 4.2% of isolates, respectively. Whole-genome sequencing (WGS) identified four distinct species within the B. cereus group, with 21 isolates closely related to B. cereus sensu stricte, 25 to B. mosaicus, 2 to B. toyonensis, and 1 to B. mycoides. Three novel sequence types (STs 3297, 3298, 3299) were discovered. Antibiotic resistance genes were common, with 100% of isolates carrying beta-lactam resistance genes. Fosfomycin (80%), vancomycin (8%), streptothricin (6%), tetracycline (4%), and macrolide resistance (2%) genes were also detected. These results highlight the genetic diversity and antibiotic resistance potential of B. cereus sensu lato strains in Polish food products. Full article

The accumulation of high amounts of plastic waste in the environment has raised ecological and health concerns, particularly in croplands, and biological degradation presents a promising approach for the sustainable treatment of this issue. In this study, a polyvinyl chloride (PVC)-degrading bacterium was isolated from farmland soil samples attached to waste plastic, utilizing PVC as the sole carbon source. The circular chromosome of the strain Cbmb3, with a length of 5,768,926 bp, was subsequently sequenced. The average GC content was determined to be 35.45%, and a total of 5835 open reading frames were identified. The strain Cbmb3 was designated as Bacillus toyonensis based on phylogenomic analyses and genomic characteristics. The bioinformatic analysis of the Cbmb3 genome revealed putative genes encoding essential enzymes involved in PVC degradation. Additionally, the potential genomic characteristics associated with phytoprobiotic effects, such as the synthesis of indole acetic acid and secondary metabolite synthesis, were also revealed. Overall, the present study provides the first complete genome of Bacillus toyonensis with PVC-degrading properties, suggesting that Cbmb3 is a potential strain for PVC bioremediation and application. Full article

Climate change has increased variable weather patterns that affect plants. To address these issues, we developed a microbial biocontrol agent against drought stress in kimchi cabbage (Brassica rapa L. subsp. pekinensis). We selected three bacterial strains (Leifsonia sp. CS9, Bacillus toyonensis TSJ7, and Lysinibacillus capsici TT41) because they showed a survival rate of up to 50% and good growth rate when treated with 30% PEG 6000. The three strains were treated with kimchi cabbage to confirm their enhanced drought stress resistance under non-watering conditions. Among the three strains, the TT41 treated group showed a significant increase in various plant parameters compared with the negative control on the 7th day. We performed extensive profiling of primary and secondary metabolites from kimchi cabbage and the TT41 strain. Multivariate and pathway analyses revealed that only the TT41 group clustered with the well-watered group and showed almost the same metabolome on the 7th day. When treated with TT41, lactic acid was identified as an indicator metabolite that significantly improved drought stress tolerance. Furthermore, lactic acid treatment effectively induced drought stress tolerance in kimchi cabbage, similar to that achieved with the TT41 strain. Full article

Fungal infections currently pose a real threat to human lives. In the current study, soil bacterial isolates were screened for the production of antifungal compounds to combat human fungal pathogens. Notably, the bacterial F1 isolate exhibited antimycotic action towards the Candida albicans ATCC 10231 and Aspergillus niger clinical isolates. By employing phenotypic and molecular techniques, we identified the F1 isolate as the Bacillus toyonensis isolate OQ071612. The purified extract showed stability within a pH range of 6–7 and at temperatures of up to 50 °C. It demonstrated potential antifungal activity in the presence of various surfactants, detergents, and enzymes. The purified extract was identified as 6-methoxy-1H-Indole-2-carboxylic acid using advanced spectroscopic techniques. To optimize the antifungal metabolite production, we utilized response surface methodology (RSM) with a face-centered central composite design, considering nutritional and environmental variables. The optimal conditions were as follows: starch (5 g/L), peptone (5 g/L), agitation rate of 150 rpm, pH 6, and 40 °C temperature. A confirmatory experiment validated the accuracy of the optimization process, resulting in an approximately 3.49-fold increase in production. This is the first documented report on the production and characterization of 6-methoxy-1H-Indole-2-carboxylic acid (MICA) antifungal metabolite from Bacillus toyonensis. Full article

Bacillus species produce different classes of antimicrobial and antioxidant substances: peptides or proteins with different structural compositions and molecular masses and a broad range of volatile organic compounds (VOCs), some of which may serve as biomarkers for microorganism identification. The aim of this study is the identification of biologically active compounds synthesized by five Bacillus species using gas chromatography coupled to mass spectrometry (GC–MS). The current study profoundly enhances the knowledge of antibacterial and antioxidant metabolites ensuring the unambiguous identification of VOCs produced by some Bacillus species, which were isolated from vegetable samples of potato, carrot, and tomato. Phylogenetic and biochemical studies were used to identify the bacterial isolates after culturing. Phylogenetic analysis proved that five bacterial isolates BSS12, BSS13, BSS16, BSS21, and BSS25 showed 99% nucleotide sequence similarities with Bacillus safensis AS-08, Bacillus cereus WAB2133, Bacillus acidiproducens NiuFun, Bacillus toyonesis FORT 102, and Bacillus thuringiensis F3, respectively. The crude extract was prepared from bacterial isolates to assess the antibiotic resistance potency and the antimicrobial potential against various targeted multidrug-resistant strains, including yeast strains such as Candida albicans, Candida krusei, and bacterial strains of Enterococcus hirae, Escherichia coli, Klebsiella aerogenes, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus group B, Streptococcus mutans, Shigella sonnei, Salmonella enteritidis, Serratia marcescens, Pseudomonas aeruginosa, and Proteus vulgaris. GC–MS analysis of bacterial strains found that VOCs from Bacillus species come in a variety of chemical forms, such as ketones, alcohols, terpenoids, alkenes, etc. Overall, 69 volatile organic compounds were identified from five Bacillus species, and all five were found to share different chemical classes of volatile organic components, which have a variety of pharmacological applications. However, eight antibacterial compounds with different concentrations were commonly found in all five species: acetoin, acetic acid, butanoic acid, 2-methyl-, oxime-, methoxy-phenyl, phenol, 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester, nonanoic acid, and hexadecanoic acid, methyl. The present study has demonstrated that bacterial isolates BSS25, BSS21, and BSS16 display potent inhibitory effects against Candida albicans, while BSS25, BSS21, and BSS13 exhibit the ability to restrain the growth and activity of Candida krusei. Notably, BSS25 and BSS21 are the only isolates that demonstrate substantial inhibitory activity against Klebsiella aerogenes. This disparity in inhibitory effects could be attributed to the higher concentrations of acetoin in BSS25 and BSS21, whereas BSS16 and BSS13 have relatively elevated levels of butanoic acid, 2-methyl-. Certainly, the presence of acetoin and butanoic acid, 2-methyl-, contributes to the enhanced antibacterial potential of these bacterial strains, in conjunction with other organic volatile compounds and peptides, among other factors. The biology and physiology of Bacillus can be better understood using these results, which can also be used to create novel biotechnological procedures and applications. Moreover, because of its exceptional ability to synthesize and produce a variety of different antibacterial compounds, Bacillus species can serve as natural and universal carriers for antibiotic compounds in the form of probiotic cultures and strains to fight different pathogens, including mycobacteria. Full article

Unexpected atypical isolates of Bacillus cereus s.l. occasionally challenge conventional microbiology and even the most advanced techniques for anthrax detection. For anticipating and gaining trust, 65 isolates of Bacillus cereus s.l. of diverse origin were sequenced and characterized. The BTyper3 tool was used for assignation to genomospecies B. mosaicus (34), B. cereus s.s (29) and B. toyonensis (2), as well as virulence factors and toxin profiling. None of them carried any capsule or anthrax-toxin genes. All harbored the non-hemolytic toxin nheABC and sphygomyelinase spH genes, whereas 41 (63%), 30 (46%), 11 (17%) and 6 (9%) isolates harbored cytK-2, hblABCD, cesABCD and at least one insecticidal toxin gene, respectively. Matrix-assisted laser desorption ionization-time of flight mass spectrometry confirmed the production of cereulide (ces genes). Phylogeny inferred from single-nucleotide polymorphisms positioned isolates relative to the B. anthracis lineage. One isolate (BC38B) was of particular interest as it appeared to be the closest B. anthracis neighbor described so far. It harbored a large plasmid similar to other previously described B. cereus s.l. megaplasmids and at a lower extent to pXO1. Whereas bacterial collection is enriched, these high-quality public genetic data offer additional knowledge for better risk assessment using future NGS-based technologies of detection. Full article

This study was conducted to investigate the impact of probiotic supplementation using Lactobacillus plantarum DSA 20174 and/or Bacillus toyonensis ATCC 55050 on growth performance, blood parameters, hematological measures, and fecal microbiota in pre-weaning Holstein calves. Thirty-two four-day-old male calves with a similar genetic background, weighing an average of 38.27 ± 0.12 kg, were randomly assigned to four groups. The groups consisted of a control group (CON) without supplementation, a group receiving B. toyonensis (BT) at 3 × 10⁹ cfu/calf/day, a group receiving L. plantarum (LP) at 1 × 10¹⁰ cfu/calf/day, and a group receiving a combination of LP and BT (LP + BT) at half the dosage for each. The study found that calves supplemented with LP and LP + BT experienced significant improvements in average daily gain and final body weight compared to the control group. The LP + BT group showed the most positive effects on TDMI, starter intake, and CP intake. RBC counts tended to be higher in the probiotic groups, with the LP + BT group having the highest values. The LP + BT group also had higher total protein, albumin, globulin, and hematocrit concentrations. All probiotic groups showed higher serum IgG concentrations. Probiotic supplementation led to increased total bacterial count and decreased levels of E. coli, salmonella, and clostridium. The LP + BT group had a significant decrease in coliform count, while both LP and LP + BT groups had increased Lactobacillus populations. In conclusion, LP + BT probiotic supplement showed the most beneficial effects on growth, feed efficiency, blood constituents, and modulation of fecal microbiota composition. Full article

Fungal pathogens are the main causal agents of postharvest diseases of fruits and vegetables. To prevent this problem and avoid the use of harmful chemical fungicides, safer and greener alternatives have been sought. One of these alternatives is the use of plant-growth-promoting bacteria (PGPB). In this study, we evaluated in vitro four well-known PGPB strains (Pseudomonas fluorescens UM270, Bacillus toyonensis COPE52, Bacillus sp. E25, and Bacillus thuringiensis CR71) for their biocontrol potential against nineteen postharvest fungal pathogens. In vivo assays were also performed, and bacterial cells were inoculated on harvested strawberries and grapes with the pathogens Botrytis cinerea, Alternaria alternata, and Fusarium brachygibbosum to evaluate loss of firmness and disease incidence. Our results show that the four strains antagonized fungi in direct and indirect confrontation assays. Stronger antagonism was observed by the action of diffusible metabolites (DMs) compared to volatile organic compound (VOC) activity. All PGPB significantly improved the fruit firmness and reduced disease incidence caused by the fungal pathogens tested. However, strain UM270 showed excellent biocontrol activity, reducing the disease incidence of Fusarium brachygibbosum, Botrytis cinerea, and Alternaria alternata on strawberry fruits by 60%, 55%, and 65%, respectively. Diffusible antifungals and VOCs such as 2,4-diacetyl phloroglucinol, siderophores, auxins, fengycins, and N, N-dimethyl-hexadecyl amine, among others, might be responsible for the beneficial activities observed. These results suggest excellent biocontrol activities to inhibit postharvest pathogenic fungi and improve harvested fruit quality. Full article

During 2022, intense heat waves, together with particularly extreme dry conditions, created a propitious scenario for wildfires, resulting in the area of vegetation consumed in Europe doubling. Mediterranean countries have been particularly affected, reaching 293,155 hectares in Spain, the worst data in the last 15 years. The effects on the vegetation and the soil are devastating, so knowing the recovery factors is essential for after-fire management. Resilient microorganisms play a fundamental role in rapid nutrient recycling, soil structure, and plant colonization in fire-affected soils. In this present work, we have studied emergent microbial communities in the case of the Los Guájares (Granada, Spain) fire, one of the most extensive of the year, to evaluate their role in the recovery of soil and vegetation cover. We aim to discern which are the main actors in order to formulate a new treatment that helps in the ecosystem recovery. Thus, we have found the relevant loss in phosphorous and potassium solubilizers, as well as siderophores or biofilm producers. Here, we decided to use the strains Pseudomonas koreensis AC, Peribacillus frigoritolerans CB, Pseudomonas fluorescens DC, Paenibacillus lautus C, Bacillus toyonensis CD, and Paenarthrobacter nitroguajacolicus AI as a consortium, as they showed most of the capacities required in a regenerative treatment. On the other hand, the microcosm test showed an enhanced pattern of germination of the emerging model plant, Bituminaria bituminosa, as well as a more aggregated structure for soil. This new approach can create a relevant approach in order to recover fire-affected soils in the future. Full article

Causing major health and ecological disturbances, polychlorinated biphenyls (PCBs) are persistent organic pollutants still recovered all over the world. Microbial PCB biotransformation is a promising technique for depollution, but the involved molecular mechanisms remain misunderstood. Ligninolytic enzymes are suspected to be involved in many PCB transformations, but their assessments remain scarce. To further inventory the capabilities of microbes to transform PCBs through their ligninolytic enzymes, we investigated the role of oxidase and peroxidase among a set of microorganisms isolated from a historically PCB-contaminated site. Among 29 isolated fungi and 17 bacteria, this work reports for the first time the PCB-transforming capabilities from fungi affiliated to Didymella, Dothiora, Ilyonectria, Naganishia, Rhodoturula, Solicoccozyma, Thelebolus and Truncatella genera and bacteria affiliated to Peribacillus frigotolerans, Peribacillus muralis, Bacillus mycoides, Bacillus cereus, Bacillus toyonensis, Pseudarthrobacter sp., Pseudomonas chlororaphis, Erwinia aphidicola and Chryseobacterium defluvii. In the same way, this is the first report of fungal isolates affiliated to the Dothiora maculans specie and Cladosporium genus that displayed oxidase (putatively laccase) and peroxidase activity, respectively, enhanced in the presence of PCBs (more than 4-fold and 20-fold, respectively, compared to controls). Based on these results, the observed activities are suspected to be involved in PCB transformation. Full article