Exopolysaccharides (EPSs) produced by Bacillus species have recently emerged as promising commercial antioxidants in various industries, such as pharmaceutics and biomedicine. However, little is known about EPS production and function from Bacillus velezensis so far. In the present study, the effect of sugar sources on EPS production by B. velezensis VTX20 and the genetic biosynthesis, characteristics, and antioxidant activity of the resulting EPS were evaluated. The strain VTX20 produced the maximum EPS yield of 75.5 ± 4.8 g/L from an initial 200 g/L of sucrose after a 48-h cultivation. Through genomic analysis, ls-levB operon was found, for the first time, to be responsible for the levan-type EPS production in B. velezensis. Biochemical and structural characterization further confirmed the majority of levan, followed by an extremely low level of dextran biopolymer. The water solubility index and water holding capacity of the EPSs were 81.9 ± 3.4% and 100.2 ± 3.4%, respectively. In vitro antioxidant activity analyses showed strong scavenging activity for 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical values of 40.1–64.0% and 16.0–40%, respectively. These findings shed light on the EPS biosynthesis of B. velezensis at both structural and genetic levels and the potential application of EPS as a natural antioxidant for pharmaceutical and biomedical industries.

Background: Fungal stem end rot disease of pitaya caused by Alternaria alternata is one of the most destructive diseases in Binh Thuan province, Vietnam. This study aimed to assess the antagonistic effects of some endophytic bacteria isolated from the weed plant (Echinochloa colonum) against A. alternata. Results: A total of 19 endophytic bacteria were isolated and 5 of them presented in vitro antagonistic activity against A. alternata. Of five, strain EC80 significantly inhibited the pathogenic growth with a mean inhibition diameter of 11.88 ± 0.08 mm, while the other four (C79, EC83, EC90, and EC97) showed a weak inhibition. Interestingly, the combination of EC79 and EC80 reduced more biomass of pathogenic fungi than the single one did. EC79 showed positive results for amylase, indole acetic acid (IAA), and biofilm production, whereas EC80 presented positive capabilities for IAA and biofilm production and a negative one for amylase production. In addition, the combined filtrate of EC79 and EC80 presented non-antifungal activity on biocontrol tests in vitro, indicating that bacteria cells played a role in defending against the pathogen. Moreover, both isolates EC79 and EC80 significantly increased seedling biomass than the control. Conclusions: The results suggest that those two strains in combination had the potential to be used as a biocontrol agent against A. alternata. More studies should be done in the future to evaluate their efficiency under the field conditions.

Purpose: To date, a total of 13 probiotic Bacillus species are considered as a Generally Recognized as Safe organism (GRAS) approved by the US Federal Food, Drug, and Cosmetic Act (FDCA), which are used for food and feed additives. However, Bacillus velezensis is not considered as a probiotic candidate in swine farming due to a lack of genetic basis of probiotic action-related traits. Therefore, the present study was undertaken to exploit the genetic basis underlying the probiotic traits of B. velezensis VTX9. Methods: The genome sequencing of B. velezensis VTX9 was performed on a PacBio Sequel platform. The probiotic properties including biosafety, antioxidative capacity, and riboflavin and exopolysaccharide production were evaluated by using genotypic and phenotypic analysis. The secondary metabolite potentials were also predicted. Results: Strain VTX9 isolated from swine feces proved some probiotic properties including resistance to 3 mM H₂O₂, 0.6 mM bile salt, low pH, and antipathogenic activity. The complete genome of B. velezensis VTX9 consists of a 3,985,800 bp chromosome that housed 3736 protein-coding genes and 5 plasmids with the size ranging from 7261 to 20,007 bp. Genome analysis revealed no functional genes encoding enterotoxins and transferable antibiotic resistance, which confirmed the safety of VTX9. A total of 82 genes involved in gastrointestinal stress tolerance were predicted, which has not been reported previously. The maximum production of riboflavin reached 769 ± 7.5 ng/ml in LB medium after 72 h, which was in agreement with the complete de novo riboflavin biosynthetic pathway exploited for the first time in the B. velezensis genome. Antagonistic activity against pathogenic bacteria was attributed to 10 secondary metabolites clusters. The presence of a large gene cluster involved in biosynthesis of exopolysaccharides underscored further the adhesion and biofilm-forming capabilities of VTX9 in swine intestines. Conclusion: Our results revealed for the first time that B. velezensis VTX9 has the potential to be a probiotic candidate. The information provided here on the genome of B. velezensis VTX9 opens new opportunities for using B. velezensis as a feed additive for swine farming in the future.

The present study aimed to evaluate the synergistic effects of the crude ethyl acetate extract (CEAE) from endophytic actinomycete MPT42 and essential oil (EO) of the same host plant Litsea cubeba. The isolate MPT42, exhibiting broad-spectrum antimicrobial activities and harboring all three antibiotic-related biosynthetic genes pks-I, pks-II, and nrps, was identified as Streptomycete griseorubens based on an analysis of the morphology, physiology, and 16S rDNA sequence. Minimum inhibitory concentrations (MICs) and the fractional inhibitory concentration index were used to estimate the synergistic effects of various combined ratios between CEAE or antibiotics (erythromycin, vancomycin) and EO toward 13 microbial strains including pathogens. L. cubeba fruit EO, showing the main chemical constituents of 36.0% citral, 29.6% carveol, and 20.5% limonene, revealed an active-low against tested microbes (MICs ≥ 600 μg/mL). The CEAE of S. griseorubens culture exhibited moderate–strong antimicrobial activities against microbes (MICs = 80–600 μg/mL). Analysis of the mechanism of action of EO on Escherichia coli ATCC 25922 found that bacterial cells were dead after 7 h of the EO treatment at 1 MIC (5.5 mg/mL), where 62% cells were permeabilized after 2 h and 3% of them were filament (length ≥ 6 μm). Combinations of CEAE, erythromycin, or vancomycin with EO led to significant synergistic antimicrobial effects against microbes with 4–16 fold reduction in MIC values when compared to their single use. Interestingly, the vancomycin–EO combinations exhibited a strong synergistic effect against five Gram-negative bacterial species. This could assume that the synergy was possibly due to increasing the cell membrane permeability by the EO acting on the bacterial cells, which allows the uptake and diffusion of antimicrobial substances inside the cell easily. These findings in the present study therefore propose a possible alternative to combat the emergence of multidrug-resistant microbes in veterinary and clinics.