Search Results (274)

Triticale (Triticosecale Wittmack) is a versatile forage crop valued for its high yield, balanced nutrition, and environmental adaptability. However, the dough-stage triricale has higher dry matter and starch content but lower water-soluble carbohydrate levels than earlier stages, posing fermentation challenges that may impair silage quality. This study aimed to investigate the effects of lactic acid bacteria inoculation on the fermentation quality, bacterial community, and metabolome of whole-plant triticale silage at the dough stage. Fresh triticale was ensiled for 30 days without or with an inoculant containing Lactiplantibacillus plantarum and Streptococcus bovis. Fermentation quality, bacterial succession, and metabolic profiles were analyzed at multiple time points. Inoculation significantly improved fermentation quality, characterized by a rapid pH drop, increased lactic acid production, and better preservation of fiber components. Microbial analysis revealed that inoculation successfully established Lactobacillus as the dominant genus while suppressing spoilage bacteria like Enterobacter and Clostridium. Metabolomic analysis on day 30 identified numerous differential metabolites, indicating that inoculation primarily altered pathways related to amino acid and purine metabolism. In conclusion, inoculating dough-stage triticale with this LAB combination effectively directs the fermentation trajectory. It enhances silage quality not only by optimizing organic acid profiles and microbial succession but also by modulating key metabolic pathways, ultimately leading to improved nutrient preservation. Full article

Soil degradation exerts profound impacts on soil ecological functions, global food security, and human development, making the development of effective technologies to mitigate degradation a critical research focus. Microorganisms play a leading role in rehabilitating degraded land, improving soil hydraulic properties, and enhancing soil structural stability. Mosses contribute to soil particle fixation through their unique rhizoid structures; however, the mechanisms underlying their interactions in mixed inoculation remain unclear. Therefore, this study addresses soil and water loss caused by rainfall erosion in the cold black soil region. We conducted controlled laboratory experiments cultivating Bacillus subtilis and cold-adapted moss species, evaluating the erosion mitigation effects of different biological treatments under gradient slopes (3°, 6°, 9°) and rainfall intensities (70 mm h⁻¹, 120 mm h⁻¹), and elucidating their carbon-based structural reinforcement mechanism. The results indicated that compared to the control group, Treatment C significantly increased the mean weight diameter (MWD) and geometric mean diameter (GMD) of soil aggregates by 121.6% and 76.75%, respectively. In separate simulated rainfall events at 70 mm h⁻¹ and 120 mm h⁻¹, Treatment C reduced soil loss by 95.70% and 96.75% and decreased runoff by 38.31% and 67.21%, respectively. Crucially, the dissolved organic carbon (DOC) loss rate in Treatment C was only 21.98%, significantly lower than that in Treatment A (32.32%), Treatment B (22.22%), and the control group (51.07%)—representing a 59.41% reduction compared to the control. This demonstrates the following: (1) Bacillus subtilis enhances microbial metabolism, driving carbon conversion into stable pools, while mosses reduce carbon leaching via physical barriers, synergistically forming a dual “carbon protection–structural reinforcement” barrier. (2) The combined inoculation optimizes soil structure by increasing the proportion of large soil particles and enhancing aggregate stability, effectively suppressing soil loss even under extreme rainfall erosion. This study elucidates, for the first time, the biological pathway through which microbe–moss interactions achieve synergistic carbon sequestration and erosion resistance by regulating aggregate formation and pore water dynamics. It provides a scalable “carbon–structure”-optimized biotechnology system (co-inoculation of Bacillus subtilis and moss) for the ecological restoration of the cold black soil region. Full article

Soil salinization has become a major obstacle to global agricultural sustainability. While microbial inoculants show promise for remediation, the functional coordination between Trichoderma and PGPR in saline alkali rhizospheres requires systematic investigation. Pot studies demonstrated that while individual inoculations of Trichoderma longibrachiatum (M) or Bacillus aryabhattai (A2) moderately improved rice growth and soil properties, their co-inoculation (A2 + M) synergistically enhanced stress tolerance and nutrient availability—increasing available nitrogen (AN +28.02%), phosphorus (AP +11.55%), and potassium (AK +8.26%) more than either strain alone, while more effectively mitigating salinity (EC −5.54%) and alkalinity (pH −0.13 units). High-throughput sequencing further revealed that the A2 + M treatment reshaped the rhizosphere microbiome, uniquely enriching beneficial taxa (e.g., Actinomycetota [+9.68%], Ascomycota [+50.58%], Chytridiomycota [+152.43%]), and plant-growth-promoting genera (e.g., Sphingomonas, Trichoderma), while drastically reducing saline-alkali-adapted Basidiomycota (−87.96%). Further analysis identified soil organic matter (SOM), AN, and AP as key drivers for the enrichment of Chytridiomycota and Actinomycetota, whereas pH and EC showed positive correlations with Mortierellomycota, Aphelidiomycota, unclassified_k__Fungi, and Basidiomycota. Collectively, the co-inoculation of Trichoderma and PGPR strains enhanced soil microbiome structure and mitigated saline alkali stress in rice seedlings. These findings demonstrate the potential of microbial consortia as an effective bio-strategy for saline alkali soil amelioration. Full article

Grapevine (Vitis vinifera) is a key crop in Mediterranean agriculture, now increasingly threatened by Xylella fastidiosa subsp. Fastidiosa (Xff), the causal agent of Pierce’s disease. This study investigated: (1) the diversity of culturable fungal endophytes in the xylem sap of naturally Xff-infected grapevines, and (2) the interaction between Xff and the pathogenic fungus Sclerotinia sclerotiorum identified in the sap. The xylem sap was collected from Cabernet Sauvignon vines in Mallorca, Spain, and fungal communities were characterized using culture-dependent methods. Both beneficial fungi (e.g., Aureobasidium pullulans, Rhodotorula mucilaginosa) and pathogenic species (e.g., S. sclerotiorum, Cladosporium sp., Alternaria alternata, and the Phoma complex) were isolated from both Xff-positive and Xff-negative plants, indicating similar community profiles. Although limited by small sample size, these findings offer preliminary evidence of complex ecological interactions between Xff and the xylem-associated mycobiota, with potential implications for grapevine health and disease development under varying environmental and management conditions. Further experiments under controlled conditions revealed that grapevines co-inoculated with Xff and S. sclerotiorum showed increased disease severity, suggesting a synergistic interaction. These preliminary results highlight the complex interplay between Xff and the fungal endophytic microbiome, which may modulate grapevine susceptibility depending on environmental and management conditions. Full article

Drought stress significantly hinders the cultivation of medicinal plants such as licorice (Glycyrrhiza uralensis), valued for its bioactive compounds, glycyrrhizin, and liquiritin. This study aims to investigate how co-inoculation with arbuscular mycorrhizal fungus Rhizophagus irregularis and Trichoderma harzianum can enhance licorice drought tolerance and secondary metabolite production, providing insights for sustainable agriculture in arid regions. The results demonstrate that inoculation with R. irregularis significantly improved biomass, drought stress tolerance, and increased glycyrrhizin and liquiritin concentrations by 29.9% and 3.3-fold, respectively, particularly under drought conditions. Co-inoculation with T. harzianum further boosted glycyrrhizin yield by 93.7%, indicating a synergistic relationship between the two microbes. The expression of key biosynthetic genes, including squalene synthase (SQS1) for glycyrrhizin and chalcone synthase (CHS) for liquiritin, was significantly upregulated, enhancing water use efficiency and the biosynthesis of secondary metabolites. Nutrient analysis showed improved phosphorus uptake, alongside reduced root carbon and nitrogen concentrations, leading to greater nutrient utilization efficiency. These findings suggest that co-inoculating R. irregularis and T. harzianum is a promising approach to improving licorice growth and medicinal quality under drought stress, with broad applications for sustainable crop management. Full article

Background/Objectives: Alternative therapies for urinary tract infections (UTIs) have been explored, but their efficacy remains inconsistent. With rising antibiotic resistance, this study aimed to evaluate simplified postbiotic formulations derived from heat-killed probiotics for long-term protection against primary and recurrent UTIs in a murine model. Methods: We compared a multi-strain (seven-strain) versus a single-strain postbiotic in preventing Escherichia coli-induced UTIs and recurrent polymicrobial UTIs, assessed protection persistence after treatment discontinuation, and established a novel sustained UTI model via intravesical co-inoculation of three uropathogens. Mice were allocated to three experimental groups: a placebo group (PBS), Postbiotic I group (a seven-strain heat-killed probiotic formulation), and Postbiotic II group (a single-strain heat-killed probiotic). After two weeks of treatment, mice were challenged with uropathogenic E. coli (UPEC) and treated for seven days. Following a 14-day washout and bacterial clearance, they were rechallenged with multidrug-resistant UPEC, Klebsiella pneumoniae, and Staphylococcus pseudintermedius. Results: Both postbiotics significantly accelerated bacterial clearance in primary UTIs (p < 0.05). In recurrent UTIs, placebo-treated mice exhibited persistent bacteriuria, while Postbiotic I maintained a significantly higher sterile urine rate (50–80%, p < 0.01) post-treatment. Histopathological analysis confirmed reduced bladder and kidney inflammation (p < 0.05) with Postbiotic I. Conclusions: These findings demonstrate the superior efficacy of Postbiotic I in mitigating UTIs, with sustained protection post-treatment, supporting its potential as a long-term, non-antibiotic strategy. Additionally, our reproducible chronic UTI model, achieved through the co-inoculation of three uropathogens, provides a valuable tool for future research on chronic UTI pathogenesis and treatment. Full article

Although drunken horse grass (Achnatherum inebrians) can be simultaneously infected by the foliar endophyte Epichloë gansuensis and colonized by Bacillus subtilis, it remains unclear whether Epichloë endophyte symbiosis influences B. subtilis colonization, as well as how their interaction affects nitrogen fixation and assimilation. The purpose of the present study was to investigate whether E. gansuensis endophyte infection facilitates the colonization of B. subtilis in the roots of host plants, with a focus on understanding the interaction effects of the E. gansuensis endophyte and B. subtilis on plant growth and nutrient absorption. In this study, we measured the colony growth rate of B. subtilis LZU7 when co-cultured with E. gansuensis strains. In addition to an in vitro test, we investigated the root colonization of Epichloë endophyte-infected plants (E+) and Epichloë endophyte-free plants (E−) with the GFP-tagged B. subtilis LZU7 in an inoculation test. Furthermore, we evaluated the interactions between E. gansuensis endophyte symbiosis and B. subtilis LZU7 colonization on the dry weight, nitrogen fixation, nitrogen converting-enzyme activity, and nutrients for E+ and E− plants by labeling with ¹⁵N₂. The results showed that the growth rates of B. subtilis LZU7 were altered and increased in a co-culture with the E. gansuensis endophyte. A significantly greater colonization of GFP-tagged B. subtilis LZU7 was detected in the roots of E+ plants compared with the roots of E− plants, suggesting that E. gansuensis endophyte symbiosis enhances the colonization of beneficial microorganisms. The combination of E. gansuensis endophyte symbiosis and B. subtilis LZU7 inoculation significantly altered the expression of the nitrogenase (nifH) gene, thereby promoting increased biological nitrogen fixation (BNF). The E. gansuensis endophyte infection and inoculation with B. subtilis LZU7 significantly increased δ15NAir in plants. Co-inoculation with the E. gansuensis endophyte and B. subtilis LZU7 significantly elevated NH₄⁺ accumulation in the roots, depleted the NH₄⁺ availability in the surrounding soil, and showed no measurable impact on the foliar NH₄⁺ content. The observed alterations in the NH₄⁺ content were linked to nitrogen-fixing microorganisms that promoted nitrogen fixation, thereby enhancing nitrogen uptake and contributing to greater biomass production in A. inebrians. Our findings highlighted the fact that a foliar symbiosis with the E. gansuensis endophyte enhances the recruitment of beneficial bacteria, and that the resulting interaction significantly impacts nitrogen fixation, assimilation, and allocation in host plants. Full article

Excessive application of a chemical fertilizer during rice cultivation leads to soil infertility and increases production costs. An alternative way to reduce over-fertilization is to partially or fully replace the fertilizer with microbes that promote the growth and production of plants. This study aimed to investigate the Maled Phai rice cultivar (Oryza sativa L.) in a field experiment with two fungi strains. Rhizophagus variabilis KS-02 and Trichoderma zelobreve PBMP16 were selected as inocula and compared with non-R. variabilis KS-02 and non-T. zelobreve PBMP16, acting as controls, one without synthetic fertilizer and one with synthetic NPK fertilizer. The field experiment was conducted in a Randomized Complete Block design with four replications. Growth and yield parameters were determined in the plant tissues and roots, and bioactive compounds were determined in the rice seeds. The results show the presence of T. zelobreve PBMP16 and R. variabilis KS-02 colonization in the plant roots at the harvest stage. A single inoculum of both R. variabilis KS-02 and T. zelobreve PBMP16 significantly increased most of the plant growth performance and yield parameters, as well as the concentrations of bioactive compounds. Remarkably, such effects were more apparent than those observed with the use of a chemical fertilizer. Thus, a single inoculum of R. variabilis KS-02 or T. zelobreve PBMP16 and the co-inoculation of both have the potential to increase the grain yield and bioactive compounds of Maled Phai under field conditions. Full article

Root-knot nematodes (RKNs; Meloidogyne incognita) pose a significant threat to tomato crops, necessitating sustainable control methods. This study investigated the inoculation efficacy of co-cultured Burkholderia vietnamiensis B418 and Trichoderma harzianum T11W compared with single-strain treatments for RKNs suppression and their influence on the structure and function of the rhizosphere microbiome. Co-inoculation with B418 + T11W achieved a 71.42% reduction in the disease index, significantly outperforming single inoculations of B418 (54.46%) and T11W (58.93%). Co-inoculation also increased plant height by 38.51% and fresh weight by 76.02% compared to the RKNs infested plants control, promoting robust tomato growth. Metagenomic analysis reveals that co-inoculation enhanced bacterial diversity, with 378 unique bacterial species and a high Shannon index, while fungal diversity decreased with Trichoderma dominance (83.31% abundance). Actinomycetota (46.42%) and Ascomycota (97.92%) were enriched in the co-inoculated rhizosphere, showing negative correlations with RKNs severity. Functional analysis indicates enriched metabolic pathways, including streptomycin and unsaturated fatty acid biosynthesis, enhancing microbial antagonism. Single inoculations altered pathways like steroid degradation (B418) and terpenoid biosynthesis (T11W), but co-inoculation uniquely optimized the rhizosphere microenvironment. These findings highlight co-inoculation with B418 + T11W effectively suppressing RKNs and fostering plant health by reshaping microbial communities and functions, offering a promising approach for sustainable agriculture. Full article

This study evaluated the antimicrobial potential of a Bacillus subtilis spore-based probiotic cocktail to reduce foodborne pathogens in both nutrient-rich laboratory media and a complex food matrix (hummus). Three common foodborne pathogens—Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium—were cultured individually in full-strength, half-strength, and quarter-strength tryptic soy broth (TSB) with or without the probiotic spores (~7 log CFU/mL). Additionally, a commercial hummus formulation was inoculated with L. monocytogenes (~3 log CFU/g) and B. subtilis spores (~7 log CFU/g) and stored at 30 °C to simulate temperature abuse. In TSB, E. coli and Salmonella grew to ~8.2 log CFU/mL in full-strength media, with no significant inhibition by the probiotics. However, L. monocytogenes showed substantial suppression: in nutrient-limited TSB, viable counts dropped below the detection limit of 1.48 log CFU/mL by 24 h in the presence of probiotics. In hummus, L. monocytogenes grew to an average of 8.22 log CFU/g in the absence of probiotics but remained significantly lower at an average of 5.03 log CFU/g when co-inoculated with B. subtilis (p < 0.05). Germination of probiotic spores was confirmed within 6 h under all conditions. These findings suggest that B. subtilis spores selectively inhibit Listeria, particularly under nutrient stress or abuse conditions. While the probiotic had limited impact on Gram-negative pathogens, its application may serve as a clean-label strategy for suppressing L. monocytogenes in ready-to-eat (RTE) foods. This dual-model approach provides insights into both mechanistic activity and practical limitations of spore probiotics in complex food matrices. Full article

Lachancea thermotolerans help increase the acidity of wines by producing L-lactic acid, which can serve as a strategy to mitigate the decrease in total acidity in wines promoted by climate change. The aim of the present paper is to test the capability of wine bioacidification of wild strains isolated from Rioja AOC. For this purpose, L. thermotolerans strains isolated from musts were used in mixed fermentation (co-inoculation and sequential inoculation) with Saccharomyces cerevisiae to determine the fermentation performance and L-lactic acid production, in both laboratory scale and pilot scale. Fermentation kinetics was evaluated, in addition to the final wine chemical composition and organoleptical properties. The results indicated that the isolated strains produced L-lactic acid; these effects were dependent on the strain and the inoculation strategy, being higher the effect in sequential inoculation (9.20 g/L) than in co-inoculation. This L-lactic acid production capacity was maintained at a pilot scale (4.65 g/L), in which the acidity increase was perceptible in the sensorial analysis, and an ethanol concentration decrease was also reported. The wine acidification depends on the appropriate selection of the strains, the inoculation procedure, the yeast adaptation to media, and competence with other yeast species present in the fermentation broth. The wild L. thermotolerans Lt97 strain could be used as a bioacidification tool for wines affected by climate change. Full article

This study investigated the beneficial effects of individual and co-inoculation with Lactobacillus delbrueckii subsp. lactis N102 and Lactobacillus sakei H1-5 on improving safety parameters, sensory characteristics, and non-volatile metabolite profiles in dry-fermented sausages. Comprehensive analyses were conducted throughout the 20-day maturation period (0, 6, 13, 16, and 20 days), including physicochemical monitoring (moisture content, malondialdehyde (MDA) levels, biogenic amine concentrations, and sodium nitrite residues); sensory evaluation (color parameters and textural properties); and ¹H NMR-based metabolomic profiling. Key findings revealed strain-specific advantages: the N102 inoculation significantly delayed lipid oxidation, achieving the lowest final MDA concentration (4.5 mg/kg) among all groups. Meanwhile, H1-5 supplementation notably improved color attributes (a*/b* ratio = 1.34). The co-inoculation strategy demonstrated synergistic effects through (1) accelerated acidification (pH 5.3 by day 6); (2) enhanced textural properties (significantly increased hardness and elasticity vs. control); (3) optimized water distribution (free water reduced to 0.56% with 64.73% immobilized water); and (4) a significant reduction in sodium nitrite residues (70% decrease) and complete elimination of phenylethylamine (total biogenic amines: 702.94 mg/kg). ¹H NMR metabolomics identified 30 non-volatile metabolites, and the co-inoculation significantly increased the amount of essential amino acids (leucine, isoleucine), flavor-related compounds (glutamic acid, succinic acid), and bioactive substances (gooseberry, creatine). These metabolites enhanced antioxidant capacity, freshness, and nutritional value. Our findings demonstrate that strategic co-cultivation of food-grade lactobacilli can synergistically enhance both the techno-functional properties and biochemical composition of fermented meat products, providing a viable approach for quality optimization in industrial applications. Full article

Citruses are one of the major fruit crops globally. Among Mediterranean citrus producers, Sicily (southern Italy) is renowned for its high-quality fresh fruit production. Phytophthora diseases are a serious issue for citrus production worldwide and Phytophthora nicotianae is a prevalent causal agent of root rot in most citrus growing areas globally and particularly in the Mediterranean region. This study reports the occurrence of Phytophthora inundata as a root pathogen of declining mature citrus trees in eastern Sicily in association with P. nicotianae. This is the first record of P. inundata on citrus in Europe and the Mediterranean region. The species was identified on the basis of a morphology and multi-gene phylogenetic analysis, which included the internal transcribed spacer, β-tubulin and cytochrome c oxidase subunit 1. Pathogenicity tests on citrus saplings showed P. inundata was a less aggressive pathogen than P. nicotianae. However, the co-inoculation of both species produced more severe symptoms than inoculation with a single species, indicating an additive effect of these two pathogens and suggesting that opportunistic secondary pathogens like P. inundata may have a crucial role in complex diseases. Full article

Co-inoculation with Saccharomyces cerevisiae and non-Saccharomyces yeasts is an effective method to improve the flavor of cider. Wickerhamomyces anomalus, known for its high ester production capacity, was evaluated in combination with S. cerevisiae to identify optimal mixed yeast inoculants for improved sensory characteristics. Three W. anomalus strains and three inoculation ratio attributes (1:5, 1:1, and 5:1) were tested to assess their impact on the physicochemical indices and sensory attributes of cider. All the strains used as starters developed fermentation-producing ciders with alcoholic degrees between 6.22 and 6.36 (% v/v). Co-inoculation with W. anomalus resulted in significantly higher ester, volatile acid, and higher alcohol levels compared to those of S. cerevisiae monocultures, increasing the complexity of fruity and floral aromas. Furthermore, the proportion of W. anomalus strains in the inoculations was positively correlated with increased aromatic esters and higher alcohols. The Sc–Wa (1:5) cider showed the highest contents of ethyl ethanoate and 3-methylbutan-1-ol, contributing to a nail polish-like aroma. Sc–Wa (1:1) yielded a higher aromatic diversity than did Sc–Wa (5:1), suggesting that co-inoculation with a ratio of 1:1 may provide an effective fermentation strategy for cider aroma enhancement. These findings offer valuable insights into how non-Saccharomyces yeasts can be effectively applied in cider co-fermentation, providing a foundation for their future use in industrial applications. Full article

This study evaluates the impact of Rhizobium tropici reinoculation and Azospirillum brasilense co-inoculation at different growth stages on the physiological quality of common bean seeds. A randomized block design was used, assessing germination, vigor, electrical conductivity, seedling length, and dry mass. Treatments T7 (co-inoculation R. tropici + A. brasilense at R5) showed the highest germination rates, indicating enhanced seed viability. The accelerated aging test revealed that T7 exhibited greater resistance to stress, presenting greater seedling vigor, whereas T10 and T11 were more susceptible. The electrical conductivity results remained stable across treatments, suggesting that cell membrane integrity was not significantly compromised. Seedling length and dry mass did not present significant variations, reinforcing the idea that early germination and vigor are primary indicators of seed quality. Canonical discriminant analysis and MANOVA confirmed significant treatment differences, highlighting the influence of inoculation strategies on seed physiology. Overall, co-inoculation with Rhizobium tropici and Azospirillum brasilense (particularly in T7) demonstrated potential to improve seed quality at lower cost, offering sustainable alternatives for optimizing agricultural production. Full article