Search Results (65)

The human gut microbiota plays a critical role in health throughout life. While fruits and vegetables are well-known sources of nutrients and prebiotics, recent studies suggest they may also contribute viable microorganisms to the gut microbiome, particularly when consumed raw. However, the impact of agricultural practices—such as the use of microbial biostimulants or exposure to salt stress—on the composition of the edible plant microbiome remains poorly understood. In this study, we performed a comprehensive metataxonomic analysis of the endophytic microbiome in the edible tissues (leaves or fruits) of lettuce (Lactuca sativa) and tomato (Solanum lycopersicum), cultivated under standard conditions with or without microbial biostimulants and salt stress. Our results show that microbial biostimulants—Priestia megaterium (PGPB) and Rhizophagus irregularis (AMF)—as well as moderate salt stress, significantly reshape the composition and diversity of endophytes in both crops. Notably, the PGPB and NaCl treatments enhanced the abundance of bacterial genera such as Pantoea, Stenotrophomonas, and Massilia, which are associated with plant health and may have probiotic potential. Salt stress also increased alpha-diversity indices and favored the presence of Firmicutes and Bacteroidota, phyla commonly linked to a healthy human gut microbiome. Agronomic inputs used in organic and conventional farming, such as microbial biostimulants or controlled salt exposure, may represent novel strategies to enhance the microbial quality of fresh produce and promote gut microbial diversity through diet. Full article

The comprehensive analysis of microbial communities reveals the unique microbial identity of different olive varieties, paving the way for new strategies in their development and commercial exploitation. In this context, the present study aimed to explore the microbial diversity and functional characteristics of Tsounati variety olives from the Monemvasia region of Peloponnese, Greece, that were naturally fermented for three months. The bacterial and fungal microbiota of both olives and brines were fingerprinted throughout the fermentation through classical microbiological analysis combined with molecular techniques. Among the 148 isolated bacteria, 85 were lactic acid bacteria (LAB), and 63 belonged to the Enterobacteriaceae family, while the 178 fungal isolates comprised 136 yeasts and 42 non-yeast or yeast-like fungi. Metataxonomic analysis confirmed the dominance of the bacterial genera Lactiplantibacillus, Leuconostoc, along with the Enterobacteriaceae family, and it revealed the presence of Coleofasciculaceae cyanobacteria mostly in olives. The dominant fungal genera were yeasts, namely Saccharomyces, Nakazawaea, and Cyberlindnera. Using the Folin–Ciocalteu assay, the average total polyphenol content of Tsounati fermented olive samples was 761.80 ± 128.87 mg gallic acid equivalents kg⁻¹ after 90 days of fermentation. The concentrations of the triterpenic, maslinic, and oleanolic acids, as determined by HPLC, remained stable throughout fermentation, with average values of 4764 and 1807 mg kg⁻¹, respectively. Finally, sensory analysis revealed the rich aromatic character of Tsounati variety, highlighting its potential to be used for Greek-style table olive production. Full article

The objective of this study was to evaluate the effect of tyrosine on behavior and ruminal meta-taxonomic profile of Altay sheep. Twelve rams with the lowest behavioral responses to humans and twelve rams with the highest behavioral responses were sorted into calm and nervous groups, respectively. Following the 2 × 2 factorial design, the rams from each group were equally assigned two treatments of a basal diet and a diet with an extra 4 g of tyrosine for 30 d. The temperament trait, growth performance, rumen fermentation parameters, ruminal epithelium antioxidant capacity, and rumen microbial composition were measured at the end of the feeding experiment. The results showed that tyrosine treatment led to a decreased number of crosses in the arena test in nervous sheep (p < 0.05). The tyrosine treatment led to increased antioxidant markers in the rumen epithelium, such as catalase, total antioxidant capacity, and GSH content in rumen epithelial tissues (p < 0.05) in calm and nervous sheep. The total volatile fatty acids and propionic acid content in the rumen were increased by tyrosine treatment in nervous and calm sheep (p < 0.05). The rumen microbial study revealed that the dominant microbial genera were Cryptobacteroides, Prevotella, Limivicinus, Quinella, UBA1711, RUG740, Sachharofermentans, Limomorpha, Soladiphilus, Flexinia, and others in both the tyrosine treatment and temperament groups. A combined effect of treatment and temperament was seen on the abundance of two microbial genera, UBA1711 and RUG740 (p < 0.05). These findings suggest that tyrosine-treated Altay sheep would experience less stress during production, resulting in reduced oxidative stress in the rumen epithelium and improved propionic acid production in the rumen compared with that of basal-diet-treated Altay sheep. Meanwhile, the effect of the dietary tyrosine treatment on ruminal microbial diversity was lower, suggesting lesser degradation of tyrosine by ruminal microbes in Altay sheep. Full article

This study explores the microbiota and metabolite profile of raw cow’s milk, analyzing the correlation between them and their potential impact on consumer health, focusing on factors like cow’s feeding regimen (conventional vs. grazing), the herd’s geographical location (coastal vs. inland), and the thermal stability of raw milk samples. To achieve this, raw milk quality was assessed through mesophilic bacterial count and thermal stability using the 70° and 80° ethanol test. Metataxonomic analysis was performed using Illumina 16S rRNA hypervariable region sequencing, and untargeted metabolomics was conducted using UHPLC-Q/TOF MS/MS, followed by multivariate correlation analysis. All samples met quality standards. A total of 1258 prokaryotic ASVs were identified, spanning 48 bacterial phyla and one archaeal phylum, with 909 genera and 349 unidentified ASVs. Additionally, 70 metabolites were identified, including essential amino acids, vitamins, and bioactive compounds with antimicrobial, anti-inflammatory, and prebiotic properties. Significant correlations between bacterial diversity and metabolite profiles were observed. Feeding regimen and geographical location influence microbial composition and metabolite abundance, while thermal stability was linked to specific metabolites like oleamide and pyridoxal. These findings suggest that microbiota-derived metabolites can enhance the nutritional and functional value of milk. Further research is needed to understand how environmental and processing factors influence these dynamics. Full article

Gas fermentation aims to fix CO₂ into higher-value compounds, such as short or medium-chain fatty acids or alcohols. In this context, the use of mixed microbial consortia presents numerous advantages, including increased resilience and adaptability. The current study aimed to improve the performance of an enriched mixed microbial population via bioaugmentation with Megasphaera sueciensis and Clostridium carboxidivorans to improve the metabolite spectrum. The initial fermentation in trickle-bed reactors mainly yielded acetate, a low-value compound. Introducing M. sueciensis, which converts acetate into higher-chain fatty acids, shifted production toward butyrate (up to 3.2 g/L) and caproate (1.1 g/L). The presence of M. sueciensis was maintained even after several media swaps, showing its ability to establish itself as a permanent part of the microbial community. Metataxonomic analysis confirmed the successful integration of M. sueciensis into the mixed culture, with it becoming a dominant member of the Veillonellaceae family. In contrast, bioaugmentation with C. carboxidivorans was unsuccessful. Although this strain is known for producing alcohols, such as butanol and hexanol, it did not significantly enhance alcohol production, as attempts to establish it within the microbial consortium were unsuccessful. Despite these mixed results, bioaugmentation with complementary microbial capabilities remains a promising strategy to improve gas fermentation efficiency. This approach may enhance the economic feasibility of industrial-scale renewable chemical production. Full article

This study evaluated the effects of adding the autochthonous cultures Limosilactobacillus mucosae CNPC007 (LM) and Lactiplantibacillus plantarum CNPC003 (LP), originally isolated from goat milk and goat cheese, respectively, on microbiological safety, microbiota composition (analyzed through 16S rRNA gene metataxonomic sequencing), and biogenic amine (BA) production in artisanal goat (coalho) cheese made from raw or pasteurized milk during 60 days of ripening at 10 °C. Six types of cheese were produced, varying in milk treatment (raw or pasteurized) and the presence or absence of LP or LM cultures. Adding either LP or LM significantly modulated the microbiota, favoring Streptococcus dominance and reducing overall bacterial diversity compared to non-inoculated cheeses. Raw milk cheeses with added autochthonous cultures exhibited a microbial profile like pasteurized cheeses, suggesting a homogenizing effect on the microbiome. Both cultures effectively reduced microbial load in raw milk cheeses after 20 days, reaching levels comparable to pasteurized cheeses by the end of ripening. Although BA concentrations increased over time, all samples remained within safe limits. Cheeses with LP addition exhibited lower BA levels, suggesting a modulating effect on their biosynthesis. Histamine concentrations were higher in raw milk cheeses with added cultures but remained well below hazardous levels. These findings suggest that incorporating either LP or LM strains is a promising strategy for enhancing the microbial safety and standardization of artisanal goat cheese while preserving its traditional characteristics. Full article

Soil health is essential for maintaining ecosystem balance, food security, and human well-being. Anthropogenic activities, such as climate change and excessive agrochemical use, have led to the degradation of soil ecosystems worldwide. Microbiome transplantation has emerged as a promising approach for restoring perturbed soils; however, direct soil transfer presents practical limitations for large-scale applications. An alternative strategy involves extracting microbial communities through soil washing processes, but its success highly depends on proper microbiota characterization and efficient extraction methods. This study evaluated a soil wash method using four different dispersant solutions (Tween-80, NaCl, sodium citrate, and sodium pyrophosphate) for their ability to extract the majority of microbial cells from Antarctic and Crop soils. The extracted microbiomes were analyzed using 16S rRNA gene metataxonomics to assess their diversity and abundance. We found that some treatments extracted a greater proportion of specific taxa, and, on the other hand, some extracted a lower proportion than the control treatment. In addition, these dispersant solutions showed the extraction of the relevant microbial community profile in soil samples, composed of multiple taxa, including beneficial bacteria for soil health. Our study aims to optimize DNA extraction methods for microbiome analyses and to explore the use of this technique in various biotechnological applications. The results provide insights into the effect of dispersant solutions on microbiome extractions. In this regard, sodium chloride could be optimal for Antarctic soils, while sodium citrate is suggested for the Crop soils. Full article

Background: Ocular infections can cause severe complications, including blindness, and distinguishing bacterial from fungal keratitis based on clinical features alone is difficult. This study compared broad-range conventional PCR and real-time PCR methods targeting the 16S rRNA gene with traditional culture for diagnosing bacterial ocular infections. Methods: We analyzed 160 ocular specimens from 111 patients, categorizing them as septic or aseptic. The results of both conventional PCR and real-time PCR methods targeting the 16S rRNA gene were compared with traditional culture outcomes. Results: Real-time PCR demonstrated higher sensitivity than conventional PCR, and receiver operating characteristic analysis determined optimal ΔCT cutoff values of −2.13 and −4.09 for septic and aseptic specimens, respectively. Delays in specimen processing significantly affected real-time PCR accuracy. The 16S rRNA meta-taxonomic analysis using nanopore sequencing only validated the PCR results when the DNA concentration was sufficient. Conclusions: Broad-range real-time PCR proved to be a valuable diagnostic tool, particularly in aseptic specimens, with greater sensitivity and specificity than conventional PCR. The established ΔCT cutoff values improved diagnostic accuracy and showed that standardized specimen collection and processing are crucial for maximizing PCR efficacy. Full article

Water stress is a major limiting factor in agriculture, particularly in the Mediterranean region, where climate change exacerbates drought conditions. Soil microbiome composition plays a crucial role in plant resilience to environmental stressors, particularly water scarcity and excess. This study examines the impact of different irrigation regimes (optimal, severe deficit, and excess) on tomato soil microbiome composition and plant physiology in a Mediterranean context. Metataxonomic profiling revealed significant shifts in microbial community structure: Proteobacteria dominated under optimal irrigation (WO), Acidobacteria under water deficit (WD), and Actinobacteria under both water deficit and excess (WE). Functional analysis indicated irrigation-induced alterations in microbial metabolic pathways, influencing nutrient cycling. Soil respiration varied, peaking in the WE condition. Plant physiological responses, including gas exchange and Proline content, were significantly affected by water stress. An inverse correlation was observed between microbial diversity and chlorophyll content, suggesting a link between plant stress responses and soil microbial composition. This study underscores for the first time the intricate relationship between water availability and microbial community dynamics, emphasizing the importance of microbiome-driven soil and plant resilience, thus showing this be a key factor in agricultural sustainability under changing climatic conditions. Full article

This study aims to contribute to the understanding of the dental pulp microbiome through metataxonomic analysis of pulp tissues from clinically healthy impacted third molars in 17 patients. Samples from coronal pulp, outer radicular dentin, and negative controls were included. Each sample was spiked with a known bacterial standard. Sequencing of the 16S rRNA V3–V4 region revealed similarity in bacterial taxonomic profiles. The bacterial DNA was detected in all pulp samples, primarily originating from reagent contaminants. Although some oral taxa appeared more abundant in pulp than in dentin or controls, contaminant species overwhelmingly dominated. Most of the extracted DNA was of human origin, rather than bacterial. Our findings question the concept of a core microbiome in healthy pulp and highlight the critical need for rigorous controls in pulpal microbiome studies. 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

Iron ore extraction can lead to significant environmental degradation, particularly due to the generation of tailings during the beneficiation process. This issue was highlighted by the B1 dam collapse in Brumadinho, Brazil, in 2019. Therefore, the study and monitoring of affected areas is essential to assess soil quality throughout the rehabilitation process, whether through natural recovery or active rehabilitation practices. Microbial indicators can serve as valuable tools to track the recovery of these areas, given their high sensitivity and rapid response to environmental changes. The aim of this study was to evaluate soil microbial indicators, such as enzyme activity, microbial biomass carbon, microbial basal respiration and microbial diversity, and to select microbial approaches for monitoring the area affected by mining tailings in Brumadinho. The results indicated that the reference area initially outperformed the affected area on all evaluated bioindicators, highlighting environmental stress in the affected zone. Over the course of the study, the two areas began to show greater similarity, suggesting a natural recovery of the soil together with the return of natural vegetation. Indicators such as microbial carbon biomass went from values close to 50 mg of C Kg of soil⁻¹ in the affected area, to around 200, statistically equal to the reference. qCO₂ also varied in the affected area to values statistically equal to those of the reference over time, variated in the first collection to 0.25 mg of C-CO₂ mg of C⁻¹ h⁻¹ in the affected area against 0.1 in the reference area; in the last collection, both areas presented values close to 0.2. Enzymatic activity had superior values in the reference area about the affected area, being urease, and arylsulfatase more sensitive to show differences between areas over time. The metataxonomic data again revealed indicator species for each environment, including genera such as Bacillus, Mycobacterium, Acidibacter, and Burkholderia representative of the reference, and the genera Ramlibacter, Sinomonas, Psedarthrobacter, and Knoellia indicators of the affected area. By the end of this study, the applicability of microbial indicators for monitoring soil microbiota and its ecosystem services was successfully demonstrated. In addition, specific microbial indicators were proposed for monitoring areas affected by iron mining tailings. Full article

Birds, fish, and marine mammals consumed by indigenous people are included in Arctic biomonitoring. However, there are still many gaps in the data on the microbiota associated with these animals. In the current study, we used high-throughput 16S rRNA gene sequencing to explore the bacterial diversity and composition in the intestines of willow ptarmigans, greater white-fronted geese, and taiga bean geese, which are widely consumed by indigenous people in the Arctic. For the first time, meta-taxonomic data have been obtained on the lungs of wild resident and migratory birds of the Russian North. The potentially pathogenic bacterial genera Helicobacter and Olsenella were found in the intestinal microbiomes of three bird species and in the lungs of willow ptarmigan. Bacteria of the genus Staphylococcus were individually identified in the intestines of willow ptarmigan, Campylobacter sp. in the intestines of taiga bean goose, and Sutterella sp. in the intestines of greater white-fronted goose as potential pathogens. The primary findings will be used to propose a next-generation sequencing scheme for monitoring both chemical and biological contaminants in the Arctic in line with One Health approach. Full article

The aim of the present study was to investigate the microbiota of processed ready-to-eat (fried or boiled) edible insects sold by street vendors at local green markets in Thailand (Bangkok and Koh Samui). To this end, samples of 4 insect species (rhino beetle adults, silkworm pupae, giant waterbugs adults, and black scorpions) were collected and analyzed through viable counting and metataxonomic analysis. Enterobacteriaceae showed counts below 1 log cfu g⁻¹ in all samples, except for black scorpions, which showed elevated counts reaching up to 4 log cfu g⁻¹. Total mesophilic aerobes counts were up to 8 log cfu g⁻¹ in all the analyzed samples. Counts below 1 log cfu g⁻¹ were observed for Escherichia coli, Staphylococcus aureus, sulfite-reducing clostridia viable cells and spores, and Bacillus cereus. All the samples showed the absence of Listeria monocytogenes and Salmonella spp. According to metataxonomic analysis, 14 taxa were consistently present across all insect samples, including Dellaglioa algida, Latilactobacillus curvatus, Latilactobacillus sakei, Acetobacteraceae, Apilactobacillus kunkeei, Bombilactobacillus spp., Enterobacteriaceae, Gilliamella spp., Lactobacillus spp., Lactobacillus apis, Streptococcus thermophilus, Lacticaseibacillus rhamnosus, Lactiplantibacillus plantarum, and Weissella spp. Minority taxa included Alcaligenes spp., Brochothrix thermosphacta, Psychrobacter spp., Staphylococcus saprophyticus, Lactobacillus melliventris, Pediococcus spp., Levilactobacillus brevis, and Snodgrassella alvi. Full article

Pneumonia caused by Pneumocystis jirovecii infection (PCP) is a potentially life-threatening illness, particularly affecting the immunocompromised. The past two decades have shown an increase in PCP incidence; however, the underlying factors that promote disease severity and fatality have yet to be fully elucidated. Recent evidence suggests that the microbiota of the respiratory tract may play a role in stimulating or repressing pulmonary inflammation, as well as the progression of both bacterial and viral pneumonia. Here, we employed 16S rRNA metataxonomic sequencing to profile the respiratory microbiota of patients with mild-moderate and severe PCP. Our results show that the upper and lower airways of PCP patients have bacterial profiles which have been associated with a pro-inflammatory response. Furthermore, we find that severe PCP is associated with lower bacterial diversity and an increase in Prevotella and a decrease in Neisseria. Functionally, severe PCP was associated with a decrease in metabolic pathways of molecules with anti-inflammatory and antimicrobial properties. To our knowledge, this is the first study showing an association of PCP severity with shifts in the respiratory microbiome and may provide some insight into which patients are more susceptible to the more severe manifestations of the disease. Full article