Pan-genome analysis identifies intersecting roles for Pseudomonas specialized metabolites in potato pathogen inhibition

Agricultural soil harbors a diverse microbiome that can form beneficial relationships with plants, including the inhibition of plant pathogens. Pseudomonas spp. are one of the most abundant bacterial genera in the soil and rhizosphere and play important roles in promoting plant health. However, the genetic determinants of this beneficial activity are only partially understood. Here, we genetically and phenotypically characterize the Pseudomonas fluorescens population in a commercial potato field, where we identify strong correlations between specialized metabolite biosynthesis and antagonism of the potato pathogens Streptomyces scabies and Phytophthora infestans. Genetic and chemical analyses identified hydrogen cyanide and cyclic lipopeptides as key specialized metabolites associated with S. scabies inhibition, which was supported by in planta biocontrol experiments. We show that a single potato field contains a hugely diverse and dynamic population of Pseudomonas bacteria, whose capacity to produce specialized metabolites is shaped both by plant colonization and defined environmental inputs.

Nutraceutical Chewing Candy Formulations Based on Acetic, Alcoholic, and Lactofermented Apple Juice Products

The aim of this study was to develop nutraceutical chewing candy (NCC) formulations based on acetic, alcoholic, and lactofermented apple juice (AJ) products. In addition, different texture-forming (gelatin, pectin) and sweetening (stevia, xylitol) agents were tested. To implement the aim of this study, combinations based on AJ, prepared from fresh and frozen apples, apple cider (C) samples (No.1, No.2, No.3, and No.4), and apple vinegar (V) were used. First, the most appropriate combination was selected by evaluating overall acceptability (OA) and emotions induced for consumers (EIC). In addition, the volatile compound (VC) profile, and physicochemical and antimicrobial characteristics of the developed combinations were analyzed. For AJ fermentation, lactic acid bacteria (LAB) strains possessing antimicrobial properties (LUHS122—L. plantarum and LUHS210—L. casei) were used. AJ prepared from frozen apples had 11.1% higher OA and 45.9%, 50.4%, and 33.3% higher fructose, glucose, and saccharose concentrations, respectively. All the tested C samples inhibited Bacillus subtilis and had an average OA of 6.6 points. Very strong positive correlations were found between AJ and C OA and the emotion ‘happy’; comparing lactofermented AJ, the highest OA was obtained for AJ fermented for 48 h with LUHS122, and a moderate positive correlation was found between AJ OA and the emotion ‘happy’ (r = 0.7617). This sample also showed the highest viable LAB count (7.59 log10 CFU mL−1) and the broadest spectrum of pathogen inhibition (inhibited 6 out of 10 tested pathogens). Further, acetic, alcoholic, and lactofermented AJ product combinations were tested. For the preparation of NCC, the combination consisting of 50 mL of AJ fermented with LUHS122 for 48 h + 50 mL C-No.3 + 2 mL V was selected because it showed the highest OA, induced a high intensity of the emotion ‘happy’ for the judges, and inhibited 8 out of 10 tested pathogens. Finally, the OA of the prepared NCC was, on average, 9.03 points. The combination of acetic, alcoholic, and lactofermented AJ products leads to the formation of a specific VC profile and increases the OA and antimicrobial activity of the products which could be successfully applied in the food and nutraceutical industries.

198 Effects of Bacillus Spp. on Pathogen Inhibition in vitro and on Growth Performance and Bacterial Shedding in Growing Finishing Pigs

Three experiments (EXP) were conducted to evaluate effectiveness of Bacillus strains on pathogen inhibition in vitro (EXP. 1 and 2) and on growing-finishing pig performance and bacteria shedding (EXP. 3). In EXP. 1, antimicrobial activity of 4 individual Bacillus strains were tested against 9 bacterial pathogens using agar diffusion cross-streak assay. In EXP. 2, a combination of 4 strains (FS4) was evaluated for antimicrobial activity against surrogate strains of the same pathogens in EXP. 1 using well-diffusion and competitive exclusion assays. There were strong inhibitory effects against Streptococcus agalactiae and Staphylococcus aureus aureus with zones of inhibition (ZOI) up to 18.1 and 13.7 mm, respectively. Moderate effects were observed against Listeria monocytogenes, Bordetella bronchiseptica, Escherichia coli, and Salmonella enterica serovar Typhimurium with ZOI up to 8.7, 8.7, 8.1, and 6.3 mm, respectively. On broth, FS4 inhibited Listeria completely (9 log10 CFU/mL) and reduced Staphylococcus by 8 log10 CFU/mL after 24h co-cultivation. In EXP. 3, 236 pigs (initial BW: 33 kg; 6–7 pigs/pen; 17 pens/treatment) were used to evaluate effects of feeding FS4 (7.5 x 104 CFU/g of feed) vs. the control (CON) diets. On d 0, 49, and at market, a pooled fresh fecal sample per pen was obtained for the enumeration of total Salmonella, E. coli, coliform, and Lactobacillus. Feeding FS4 improved (P < 0.05) BW (d 72), ADG (d 43–72 and d 0–72), and Feed:Gain (d 43–72) compared to CON. Overall, pigs fed FS4 were numerically heavier (1.1 kg) than CON pigs. FS4 reduced (P < 0.05) total coliform counts in pigs at market, and numerically reduced E. coli at d 49; however, FS4 had no impact on Lactobacillus. Our data indicate that FS4 had strong inhibitory effects against Streptococcus, Listeria, and Staphylococcus and feeding FS4 improved performance of pigs and lowered coliform shedding without impacting Lactobacillus.

Unraveling the Metabolite Signature of Endophytic Bacillus velezensis Strain Showing Defense Response Towards Fusarium oxysporum

Seedling blight, caused by the fungus Fusarium oxysporum, significantly lowers rice production globally. Earlier reports have opined that endophytic bacteria strains could be possible biocontrol agents, but the mechanistic actions involved are still unclear. Therefore, this study aimed to isolate the endophytic bacteria with high inhibitory activity and elucidate its possible mechanisms for inducing resistance by metabolomics. The results showed that mdj-36 had the strongest in vitro pathogen inhibition of F. oxysporum, while mdj-34 displayed the lowest inhibitory activity identified as Bacillus velezensis strains. Metabolomic analyses demonstrated that B. velezensis mdj-36 growth medium could produce higher organic acids, terpenes, and diterpene than B. velezensis mdj-34. Further investigation revealed that ‘secondary bile acid biosynthesis’ and ‘glycerophospholipid metabolism’ pathways played essential roles in defense response towards F. oxysporum. This study’s findings provide a credible theoretical basis for the possible use of the B. velezensis strain against rice seedling blight.

Biocontrol Traits Correlate With Resistance to Predation by Protists in Soil Pseudomonads

Root-colonizing bacteria can support plant growth and help fend off pathogens. It is clear that such bacteria benefit from plant-derived carbon, but it remains ambiguous why they invest in plant-beneficial traits. We suggest that selection via protist predation contributes to recruitment of plant-beneficial traits in rhizosphere bacteria. To this end, we examined the extent to which bacterial traits associated with pathogen inhibition coincide with resistance to protist predation. We investigated the resistance to predation of a collection of Pseudomonas spp. against a range of representative soil protists covering three eukaryotic supergroups. We then examined whether patterns of resistance to predation could be explained by functional traits related to plant growth promotion, disease suppression and root colonization success. We observed a strong correlation between resistance to predation and phytopathogen inhibition. In addition, our analysis highlighted an important contribution of lytic enzymes and motility traits to resist predation by protists. We conclude that the widespread occurrence of plant-protective traits in the rhizosphere microbiome may be driven by the evolutionary pressure for resistance against predation by protists. Protists may therefore act as microbiome regulators promoting native bacteria involved in plant protection against diseases.

Secretory Peptides as Bullets: Effector Peptides from Pathogens against Antimicrobial Peptides from Soybean

Soybean is an important crop as both human food and animal feed. However, the yield of soybean is heavily impacted by biotic stresses including insect attack and pathogen infection. Insect bites usually make the plants vulnerable to pathogen infection, which causes diseases. Fungi, oomycetes, bacteria, viruses, and nematodes are major soybean pathogens. The infection by pathogens and the defenses mounted by soybean are an interactive and dynamic process. Using fungi, oomycetes, and bacteria as examples, we will discuss the recognition of pathogens by soybean at the molecular level. In this review, we will discuss both the secretory peptides for soybean plant infection and those for pathogen inhibition. Pathogenic secretory peptides and peptides secreted by soybean and its associated microbes will be included. We will also explore the possible use of externally applied antimicrobial peptides identical to those secreted by soybean and its associated microbes as biopesticides.

ANALYSIS OF STRUCTURE AND ANTIMICROBIAL ACTIVITY OF CeO2 AND Nd2O3 NANOPARTICLES

Cerium oxide (CeO2) and Neodymium oxide (Nd2O3) nanoparticles using local content have been synthesized by precipitation method. The CeO2 and Nd2O3 nanoparticles were characterized by X-Ray Diffraction (XRD) and Fourier Transform Infrared (FTIR) to analyze the material phase and structure. The XRD spectrum shows that CeO2 and Nd2O3 nanoparticles have face-centered cubic and hexagonal, and cubic, respectively. The anti-microbial activity of CeO2 and Nd2O3 nanoparticles was analyzed by diffusion method using gram-negative bacteria (E. coli, S. aureus, P. aeruginosa), and gram-positive bacteria (S. entericatyphi, L. monocyogenes), and fungus (C. albicans). The result confirms that CeO2 and Nd2O3 nanoparticles have the capability of microbial pathogen inhibition. The CeO2 nanoparticles have the effective activities of inhibition for the microbial of S. aureus and S. entericatyphi, whereas Nd2O3 nanoparticles can inhibit the microbial of P. aeruginosa, S. entericatyphi, and L. monocyogenes.

The Potential of Fucose-Containing Sulfated Polysaccharides As Scaffolds for Biomedical Applications

Marine environments have a high quantity and diversity of sulfated polysaccharides. In coastal regions brown algae are the most abundant biomass producers and their cell walls have fucosecontaining sulfated polysaccharides (FCSP), known as fucans and/or fucoidans. These sulfated compounds have been widely researched for their biomedical properties, namely the immunomodulatory, haemostasis, pathogen inhibition, anti-inflammatory capacity, and antitumoral. These activities are probably due to their ability to mimic the carbohydrate moieties of mammalian glycosaminoglycans. Therefore, the FCSP are interesting compounds for application in health-related subjects, mainly for developing scaffolds for delivery systems or tissue regeneration. FCSP showed potential for these applications also due to their ability to form stable 3D structures with other polymers able to entrap therapeutic agents or cell and growth factors, besides their biocompatibility and biodegradability. However, for the clinical use of these biopolymers well-defined reproducible molecules are required in order to accurately establish relationships between structural features and human health applications.