Search Results (1,713)

Soybean is a critical economic, oil and industrial raw material crop, yet its production is often hindered by pathogen infection and pesticide residues. This study explored the synergistic effects of Rhizophagus intraradices and mycorrhizal helper bacteria (MHB) on AMF colonization, AMF spore density, total number of bacterial colonies, soybean growth, root rot disease index, and carbendazim residues. Hydroponic and pot experiments were conducted using a completely randomized design (CRD) with five biological replicates per treatment; after 30 days of growth, three replicates were randomly selected for all measurements. Results showed that inoculation with microbial agents, particularly co-inoculation, increased soybean biomass, reduced disease index, and decreased carbendazim residues. In the hydroponic experiment, co-inoculation increased plant height, aboveground fresh weight, and underground dry weight by 64.28%, 78.13%, and 109.09%, respectively, and decreased carbendazim residues by 71.84% relative to the carbendazim-alone group. In the pot experiment, co-inoculation reduced carbendazim residues by 81.25% and root rot disease index by 45.56% compared with the carbendazim-alone group. Correlation analysis showed a strong positive correlation (p < 0.001) between carbendazim degradation in hydroponic and pot systems, indicating stable degradation function across environments. Co-inoculation of R. intraradices and MHB synergistically promotes soybean growth, suppresses root rot, and reduces carbendazim residues, providing a theoretical basis for developing functional microbial inoculants for safe and green soybean production. Full article

This study aimed to isolate, characterize, and evaluate bacterial consortia capable of degrading the diamide insecticide cyantraniliprole in aqueous systems and to assess their bioremediation potential under environmentally relevant conditions. Four bacterial consortia, each comprising six isolates, demonstrated significant growth in mineral media containing cyantraniliprole as the sole carbon source, and the isolates were identified using conventional microbiological techniques in combination with MALDI-TOF-MS analysis. The bacterial consortia were enriched from pesticide-contaminated environments and systematically evaluated using microbiological, physiological, and analytical approaches to determine their degradation potential and environmental adaptability. The degradation performance of the consortia was systematically assessed under varying environmental parameters, including temperature, pH, salinity, and incubation time, with optimal degradation observed at 30–35 °C, pH 7.0–8.0, 0.5–5.0% NaCl, and 11 days of incubation at 150 rpm using an initial cyantraniliprole concentration of 50 mg/L. Biodegradation efficiency was further evaluated using DCPIP reduction assays, alongside measurements of biofilm formation and biomass production, indicating enhanced metabolic activity and adaptive responses under pesticide-induced stress. The consortia also exhibited the capacity to degrade structurally related diamide pesticides, including flubendiamide, chlorantraniliprole, cyclaniliprole, and fluchlordiniliprole, suggesting broad-spectrum biodegradation potential. Their performance was further validated in a simulated water microcosm system designed to mimic environmentally relevant contamination scenarios. In simulated contaminated water (60 mg/L cyantraniliprole), bacterial inoculants standardized to 10⁷ CFU/mL achieved substantial degradation after 20 days of incubation at 30 °C, as confirmed by HPLC analysis, with the six-strain consortium (T4), comprising Bacillus subtilis subsp. subtilis AZFS3, Bacillus pumilus AZFS5, Bacillus mojavensis AZFS15, Bacillus paramycoides AZFS18, Pseudomonas aeruginosa KZFS4, and Alcaligenes aquatilis KZFS11, demonstrating the highest removal efficiency (98.27%) and reducing the pesticide concentration to 1.00 mg/L, followed by consortium T3 (96.72%), which consisted of Bacillus subtilis Ht1, Bacillus subtilis Ht2, Bacillus mojavensis Ht3, Pseudomonas aeruginosa Ht4, Pseudomonas aeruginosa Ht5, and Pseudomonas aeruginosa Ht6. Residue analysis and predictive bioinformatic assessment further supported the biodegradation capacity of the selected bacterial communities and suggested the formation of simpler transformation products. Overall, the investigated bacterial consortia exhibited high degradation efficiency and environmental adaptability, highlighting their potential as effective and eco-friendly agents for the bioremediation of cyantraniliprole-contaminated water systems Full article

Introduction: Nasal colonization plays a pivotal role in Methicillin-resistant Staphylococcus aureus (MRSA) carriage and transmission, especially in healthcare settings. Asymptomatic carriers amongst healthcare workers (HCWs) may serve as an important source for inner-hospital transmission, besides personal increased risks of endogenous infections. Medical students are an often-overlooked part of medical staff which, while not typically included in statistics concerning HCWs, are associated with increased patient contact and exposure to healthcare-associated pathogens. This study aimed to assess MRSA carriage rate amongst clinical-year medical students in the largest Romanian medical university, in addition to identifying potential risk factors. Nonetheless, a methodological aim was incorporated, in order to evaluate the effect of nasal swab pre-moistening with sterile saline on MRSA retrieval rate. Materials and Methods: A cross-sectional study was conducted among clinical-year students from the ‘Carol Davila’ University of Medicine and Pharmacy in Bucharest, Romania. Participants completed a survey regarding potential risk factors and underwent nasal swab sampling, being randomly assigned to the two swab collection methods—dry swab or pre-moistened swab with sterile saline, randomization ensuring comparable baseline characteristics. Samples were inoculated on chromogenic MRSA agar media and incubated for 24–48 h at 35–37 °C. Isolates exhibiting characteristic growth further underwent coagulase testing, bacterial identification and methicillin resistance confirmation. Results: The study comprised 156 medical students, with an overall prevalence of asymptomatic MRSA nasal carriage of 5.76% (n = 9, 95% CI: 3.05–10.58%). No statistically significant associations were identified between MRSA carriage and hospital exposure. The prevalence of MRSA positive cultures was 5.00% (n = 4/80) among the conventional dry swab sampling subgroup, while the subgroup undergoing pre-moistened swab collection presented a 6.57% prevalence (n = 5/76), revealing no statistical significance (p = 0.74). Conclusions: Asymptomatic MRSA carriage among medical students in this cohort suggests the potential role of this population in intra-hospital transmission. In addition, pre-moistening the nasal swab for collection of the sample showed no statistically significant impact on MRSA recovery rates, correlating with existing literature on the topic. These findings further emphasize the need for strict adherence to infection prevention and control measures in hospitals. Full article

Respiratory disease remains a major challenge in pig production. This two-room pilot study evaluated whether room-level acoustic monitoring combined with physiological measurements could provide an early warning after an experimental Klebsiella pneumoniae challenge. Forty growing pigs balanced by sex and body weight were housed for 28 days in one control room and one challenged room (20 pigs/room; four pens/room). Challenged pigs were intranasally inoculated on days 8, 12, 16, and 20 with a culture whose dose was retrospectively verified by serial-dilution plating. Nasal and fecal samples were cultured on Klebsiella ChromoSelect agar, and colonies with expected morphology were enumerated as presumptive Klebsiella/K. pneumoniae colonies. A fine-tuned Audio Spectrogram Transformer (AST) classified five sound classes from facility-specific audio and was evaluated by group-blocked hold-out testing, five-fold group-blocked cross-validation, temporal deployment validation, and window-threshold sensitivity analysis. The model achieved hold-out macro-F1 of 0.947, five-fold macro-F1 of 0.928 ± 0.019, and 24 h deployment macro-F1 of 0.914. Presumptive nasal bacterial load was higher in challenged pigs at 1-week post-inoculation (log₁₀ 4.03 vs. 0.67). Group-size-standardized cough detections were also higher in the challenged room (54.84 vs. 36.80 detections/day), and daily coughing first exceeded the baseline threshold on day 8. Thresholds of 0.764 (control) and 1.115 (treatment) were obtained from an integrated score that included coughing, sneezing, ear temperatures, rectal temperature, and respiration rate; the treatment score and treatment–control contrast score first surpassed the threshold on day 8, and daily multimodal scores varied between groups (t = −6.636, p < 0.001). Integrated score improved discrimination of post-inoculation disturbance compared with cough detections alone (leave-one-day-out AUROC: 0.94 vs. 0.88). Because each condition was represented by one room, findings are exploratory temporal contrasts, not replicated treatment effects or a stand-alone diagnostic test. Full article

Metal-contaminated dredged sediments represent heterogeneous environmental matrices in which remediation responses are frequently constrained by elevated background metal loads and complex geochemical conditions. Within such systems, phytoremediation has been discussed as a nature-based management approach whose outcomes depend on plant biomass, internal metal allocation, and context-dependent interactions between plants and sediment. The present study evaluated whether bacterial and fungal plant growth-promoting microorganisms (PGPMs) were associated with changes in plant metal uptake and internal allocation in Sorghum bicolor L. and Cannabis sativa L. grown in dredged sediment collected from the Bega Canal. An outdoor pot experiment was conducted under environmentally relevant conditions, including bacterial and fungal inoculation treatments alongside non-inoculated controls, with plant responses to Cr, Ni, Cu, Zn, As, Cd, and Pb characterized using concentration- and mass-based uptake metrics, root–shoot partitioning, and sediment geochemical assessment based on pseudo-total concentrations and BCR sequential extraction fractions. Across treatments, plant responses were largely governed by intrinsic species traits and biomass production, while PGPM-associated effects remained modest and variable. Root-dominated metal retention and limited translocation were evident irrespective of species, consistent with a phytostabilization-type response rather than systematic extraction. Absolute metal uptake accounted for only a minor fraction of total sediment metal pools, underscoring the importance of interpreting concentration-based indices jointly with mass-based metrics when evaluating system-scale responses. Altogether, the findings indicate that under the investigated outdoor dredged sediment pot conditions, PGPM inoculation acts primarily as a context-specific modulator of plant responses rather than a driver of enhanced phytoremediation performance, reflecting the central role of intrinsic plant traits and stabilization-oriented processes in complex sediment systems. Full article

Fresh cowpeas have a limited shelf life at room temperature. Fermentation of cowpeas not only preserves their nutritional value but also prolongs their shelf life. This study categorized cowpea fermentation processes into natural and inoculated methods, focusing on analyzing physicochemical indices, acid production, and bacterial diversity throughout the cowpea fermentation process. We compared the moisture, protein content, and vitamin C levels of cowpeas. The acidification process was monitored using pH, total acid, and nitrite contents as indicators. Illumina MiSeq sequencing was employed to analyze the bacterial communities in fermented cowpeas at different fermentation stages. The experimental results indicated that during fermentation, pH, total acid content, and nitrite content all changed significantly. Lactobacillus exhibited high dominance in both natural fermentation and inoculated fermentation processes. Moreover, under inoculated fermentation conditions, its population size was significantly greater than that in natural fermentation. Analysis of bacterial community composition revealed that microbial diversity tended to decrease with prolonged fermentation time in both natural and inoculated fermentation systems. The results demonstrate that inoculation fermentation can shorten the fermentation cycle, lower nitrite levels, and confirm that lactic acid bacteria are the dominant microbial genus in vegetable fermentation. Full article

Salinization of agricultural soils is a global problem causing crop yield declines. This impact is caused by osmotic and oxidative stress, which plants often rely on endophytic bacteria to overcome. A bacterial isolate from the roots of the halophyte Plantago salsa was studied between 2024 and 2026, and its ability to increase barley tolerance to moderate salt stress was determined. Based on 16S rRNA gene sequencing (1410 bp), the isolate PS-50.1 was identified as Providencia sp. It demonstrated key plant growth-promoting properties, including indole-3-acetic acid production (21.4 mg L⁻¹) and phosphate solubilization (69.0 mg L⁻¹). The strain supported barley growth at 7% NaCl. Inoculation of barley seeds with this strain (10⁸ CFU L⁻¹) significantly reduced moderate salt stress in plants both in vitro and in a pot experiment. Inoculated plants under salinity conditions had greater shoot length (+11.6%) compared to non-inoculated; higher pre-flag leaf fresh weight; demonstrated decreased levels of prooxidants (H₂O₂ by 44.8% and malondialdehyde by 31.8%), higher proline accumulation (up to 2.0-fold), and increased antioxidant enzyme activity (catalase by 26.6% and ascorbate peroxidase by 191%). Furthermore, inoculated plants showed 9.4% higher water use efficiency and photosynthetic rate (+5.5%) under salt stress compared to uninoculated plants. These results indicate that the halophytic strain Providencia sp. PS-50.1 is a promising candidate for the development of microbial preparations aimed at increasing crop productivity under saline conditions. Full article

This study explored the effects of one mineral fertilizer and two microbial inoculants and their combined applications on soil physicochemical properties, bacterial community structure, and plant growth of Cymbidium faberi under potted cultivation, aiming to provide theoretical and technical support for the sustainable cultivation of ornamental orchids. A single-factor randomized block experiment was designed with eight treatments: control (CK), mineral sulfosulfuric acid potassium (HF), Bacillus subtilis (KC), Trichoderma harzianum (HC), mineral sulfosulfuric acid potassium + Bacillus subtilis (HK), mineral sulfosulfuric acid potassium + Trichoderma harzianum (HH), Bacillus subtilis + Trichoderma harzianum (KH), and mineral sulfosulfuric acid potassium + Bacillus subtilis + Trichoderma harzianum (HKH). Plant growth traits, soil properties, and soil bacterial community characteristics were measured. The effects of inoculant agents on Cymbidium faberi growth, soil environment, and bacterial community, as well as their interaction relationships, were systematically analyzed. The combination of three inoculants significantly promoted plant height and leaf thickness in Cymbidium faberi. Compared with CK, the relative abundance of Pseudomonadota and Bacteroidota in HH treatment increased by 6.0% and 11.0%, respectively, while the relative abundance of Acidobacteriota and Verrucomicrobiota decreased by 6.0% and 12.0%, respectively. Venn diagram analysis revealed 146 ASVs shared among all treatments. KC, HC, and HF had more unique ASVs, whereas HK and HKH had the fewest. Principal component analysis (PCA) was used to visualize differences in bacterial community structure. Significant differences among treatments were confirmed using ANOSIM. Ecological network analysis indicated predominantly positive (cooperative) associations among bacterial taxa, with HKH showing the highest proportion of positive edges, suggesting stronger bacterial cooperation. Correlation analysis showed that Patesibacteria, Acidobacterita, and Planctomycetota were significantly negatively correlated with pH and TP, while Bacteroidota, Actinomycetota, and Methylomirabilota were significantly positively correlated with pH. The Mantel analysis revealed a significant positive correlation between bacterial community composition and richness and pH. Further analysis using the structural equation model revealed that soil nutrients and bacterial communities were the main factors affecting plant growth. This study clarifies the response rules of plant growth, soil physicochemical properties and rhizosphere bacterial communities to different mineral fertilizer and microbial inoculant combinations, and provides a practical basis for the rational screening of functional inoculants and the construction of healthy rhizosphere microecosystems in Cymbidium faberi pot cultivation. Full article

Soil salinization is a challenge for global agriculture and can affect the yield of staple crops such as maize. Plant growth-promoting rhizobacteria (PGPR) are known to play a pivotal role in enhancing plant growth and stress resilience. However, no studies so far have reported plant growth-promoting (PGP) activity in members of the genus Marinobacter. In this study, a novel strain of Marinobacter sp. ZP-590, was identified as a PGPR based on a polyphasic taxonomic analysis, which was isolated from the rhizosphere soil of Tamarix chinensis Lour. Genomic analysis revealed that ZP-590 possesses 5370 protein-coding genes, including core metabolic, catalytic, and transport functions essential for bacterial survival and plant interactions, along with multiple genes potentially associated with PGP traits such as phosphate solubilization, nitrogen fixation, and the production of siderophore and exopolysaccharide (EPS), tryptophan (a prerequisite for IAA synthesis), and amylase. These genomic predictions were functionally validated through in vitro assays confirming all predicted PGP activities. Pot experiment results suggested that inoculation with ZP-590 enhanced maize growth under saline conditions. Compared to the non-inoculated controls, the treatment significantly increased root fresh weight (14.25%; p < 0.05) and stem fresh weight (125.04%; p < 0.01), while shoot height and leaf fresh weight showed no significant changes. Metabolomic profiling revealed that ZP-590 inoculation was associated with systemic metabolic changes in maize under saline conditions. A total of 394, 262, and 601 differentially accumulated metabolites in the root, stem, and leaf, respectively. These changes were characterized by a substantial up-regulation of antioxidant compounds, notably flavonoids, and changes in carbohydrate and lipid metabolism pathways. The changes in carbohydrate and lipid metabolism pathways may contribute to the supply of energy and structural components for stress adaptation. Meanwhile, the accumulation of antioxidant compounds significantly mitigated saline-induced oxidative damage by reducing the levels of superoxide anion (O₂⁻) in leaves. In this study, Marinobacter sp. ZP-590 is characterized as a PGPR that promotes maize growth under saline conditions. These findings provide a foundation for investigating the molecular mechanisms underlying the interaction between ZP-590 and maize under saline conditions. Full article

Potato (Solanum tuberosum) is a vital global non-cereal food crop severely threatened by bacterial wilt, caused by Ralstonia solanacearum(R. solanacearum). Conventional diagnostics like PCR and ELISA, though effective, are destructive and time-consuming, limiting large-scale field applications. This study investigates hyperspectral imaging (HSI) as a non-invasive, rapid, and accurate alternative for early detection and severity grading of potato bacterial wilt. Using a portable HSI system (400–1000 nm), spectral data were collected from inoculated potato plants (‘Longshu No. 7’) at 0, 24, 48, and 72 h post-inoculation, alongside disease severity assessment (grades 0–4). After comprehensive spectral preprocessing and feature band extraction via Competitivse Adaptive Reweighted Sampling (CARS), we developed two distinct sets of models: one for early detection (temporal classification) using Partial Least Squares-Discriminant Analysis (PLS-DA) and Principal Component Analysis-Linear Discriminant Analysis (PCA-LDA), and another for severity grading. The SNV + SG + MC + PLS-DA model achieved exceptional accuracy, exceeding 97% for early detection, while the MSC + SG + MC + CARS + PLS-DA model yielded >97% accuracy for severity grading. These results were supported by low misclassification rates in confusion matrices. This work establishes a robust HSI-based framework for high-throughput screening of resistant potato germplasm and advances precision agriculture strategies for bacterial wilt management. Full article

Plant growth-promoting fungi (PGPF) have shown broad potential to improve soil conditions and enhance root growth and development. However, few studies have examined the effects of exogenous PGPF inoculation on the growth of the medicinal plant Saposhnikovia divaricata and the associated changes in the rhizosphere microbiome. In this study, Aspergillus neoalliaceus MR-86 exhibited phosphate solubilization, growth in nitrogen-free medium, potassium solubilization, IAA production, and siderophore production. PCR assays did not detect the aflatoxin biosynthesis-related genes aflR, aflS, and omtA in strain MR-86. Pot trials demonstrated that inoculation with MR-86 significantly increased the plant height and root dry weight of S. divaricata by 10.32% and 21.05%, respectively (p < 0.05). In the rhizosphere, soil pH decreased, whereas soil alkaline-hydrolyzable nitrogen and available phosphorus levels, as well as the activities of protease, urease, and cellulase, increased significantly. Illumina NovaSeq sequencing revealed that MR-86 inoculation altered the soil microbial community structure and specifically enriched several microbial taxa, including Talaromyces, Subulicystidium, and Aspergillus. Moreover, MR-86 inoculation did not alter the composition of dominant bacterial and fungal phyla, but significantly modified microbial interactions and the topology of microbial networks. Correlation analysis indicated that the specific microbial taxa Subulicystidium, Aspergillus, and Talaromyces were positively associated with soil nutrient indices, enzyme activities, and plant growth parameters. Functional prediction analysis indicated that MR-86 treatment was predicted to be enriched bacterial metabolic pathways, including flavone and flavonol biosynthesis and ether lipid metabolism, and was predicted to increase the relative abundance of functional fungal groups such as ectomycorrhizal and wood-decomposing fungi. In summary, A. neoalliaceus MR-86 may contribute to improved growth of S. divaricata by enhancing nutrient availability and transformation and by modulating the structure and function of the rhizosphere microbiome. Full article

Cadmium (Cd) and polystyrene (PS) microplastic co-contamination in agricultural soils poses a potential threat to food security. Some functional microorganisms in soil can alleviate the dual stress of Cd and PS on crops. In this study, a biofilm-forming bacterium, Enterobacter sp. W5, was isolated from heavy metal-contaminated rhizosphere soil. Strain W5 exhibited Cd removal efficiency (46.3%) and strong biofilm-forming capacity (OD₅₇₀ = 5.05), and it effectively colonized PS microplastic surfaces. XPS analysis detected bacterial functional groups (C–O–C, C=O) and PS-associated signals (O–C=O), which may act synergistically in Cd²⁺ adsorption. Furthermore, XPS and XRD analyses revealed the presence of Cd-containing precipitates (including CdS, CdO, and Cd₃(PO₄)₂). In hydroponic wheat experiments, W5 inoculation alleviated Cd-PS combined stress, thus significantly promoting plant growth and reducing Cd accumulation by 22.6% in roots and by 34.2% in aboveground tissues. Subcellular distribution analysis revealed that W5 enhanced Cd retention in root cell walls, thereby limiting its translocation to active cellular compartments. Proteomic analysis identified a set of 11 consistently downregulated proteins, including A0A3B6HQ68 and A0A3B6KJV9, which were enriched in secondary metabolite biosynthesis pathways. Bioinformatic analysis suggests that these proteins may be associated with Cd stress responses, though their exact roles remain to be verified. Collectively, this study provides a valuable microbial resource and mechanistic insights into the application of biofilm-forming bacteria for mitigating combined heavy metal–microplastic pollution in agricultural systems. Full article

Arbuscular mycorrhizal fungi (AMF) can improve plant performance, but how they coordinately influence root metabolism and associated bacterial communities in sweet potato remains unclear. Here, a pot experiment was conducted to investigate the effects of Glomus intraradices inoculation on sweet potato seedlings by integrating analyses of rhizosphere soil properties, plant growth and nutrient uptake, root metabolomics, and rhizosphere and endophytic bacterial communities using 16S rRNA gene sequencing with FAPROTAX-based functional prediction. AMF inoculation significantly increased whole-plant fresh and dry biomass, potassium concentration and accumulation, and the accumulation of starch and water-soluble carbohydrates, while no significant effects were observed on dry matter rate or plant nitrogen and phosphorus concentration. In the rhizosphere, AMF reduced soil electrical conductivity and increased organic matter content without significantly affecting pH, alkali-hydrolyzable nitrogen, available phosphorus, or available potassium. Root metabolomic profiling identified 289 differential metabolites, with enrichment of phenylpropanoid biosynthesis, glycerophospholipid metabolism, porphyrin metabolism, and nucleotide metabolism, together with broad up-regulation of lipid-related metabolites. Bacterial communities showed strong compartment specificity, with the root endosphere displaying lower alpha diversity than the rhizosphere. Higher rhizosphere bacterial Shannon diversity was observed in the AMF treatment, together with compartment-dependent shifts in bacterial community composition; enrichment of endophytic taxa such as Devosia and Niastella was detected following AMF inoculation. Functional prediction further suggested niche differentiation between rhizosphere and endophytic bacteria, together with AMF-associated shifts in carbon- and nitrogen-related functions. Overall, these results suggest that G. intraradices inoculation is associated with enhanced sweet potato growth and enhanced potassium and carbohydrate accumulation in association with coordinated changes in rhizosphere conditions, root metabolism, and bacterial community assembly. Full article

Yeasts are among the intrinsic factors affecting the quality of beer due to their impact on flavor. Apart from a long and rich brewing tradition of natural fermentations, involving different yeast and bacterial species in regions such as Belgium, non-Saccharomyces yeasts have not been the preferred industrial inoculation strains in both winemaking and brewing sectors. This is mainly due to their slower fermentation rates, lower alcohol tolerance, and limited ability to complete fermentation. This review explores the brewing characteristics of Torulaspora delbrueckii strains, focusing on the aromatic profile of the produced beers, in comparison with the well-known Saccharomyces cerevisiae brewing strains. The effect of fermentation parameters, as well as the use of mono- and mixed cultures with S. cerevisiae, is discussed. Specific T. delbrueckii strains have demonstrated the ability to ferment wort sugars and produce alcohol levels of up to 4–6% (v/v). The main volatile compounds produced include higher alcohols, such as isoamyl and phenyl ethyl alcohol, as well as esters and their acid precursors. Among these, ethyl hexanoate and ethyl octanoate are particularly important due to their contribution to fruity aromas. Mixed fermentations involving T. delbrueckii and S. cerevisiae have been shown to enhance ester production, resulting in improved aromatic complexity in the final product. Full article

Live virus inoculation (LVI) is widely used for porcine reproductive and respiratory syndrome virus (PRRSV) stabilization, yet preparation practices and pathogen composition remain poorly characterized. This study aimed to evaluate variability in LVI preparation, quantify PRRSV genomic load, and detect additional swine pathogens. A survey was conducted to document LVI preparation methods, and samples were analyzed using reverse-transcription quantitative PCR (RT-qPCR) for PRRSV quantification and next-generation sequencing for PRRSV and the metagenomic characterization of additional pathogens. Among 61 LVI samples, substantial variability was observed in preparation practices and viral composition, with 31 distinct PRRSV variants identified and seven samples containing multiple strains. PRRSV RNA concentrations ranged from 10^1.69 to 2.52 × 10⁸ copies/mL. Metagenomic analysis detected a complete or near-complete genome for PRRSV, porcine parvovirus, and porcine circovirus type 2. Genome fragments of porcine sapovirus, porcine rotavirus, porcine astrovirus, and bacterial genetic material from Salmonella spp., Pseudomonas spp., Streptococcus spp., and Escherichia coli were also detected. These findings highlight substantial heterogeneity in LVI materials and encourage the use of next-generation sequencing to verify LVI PRRSV composition and screen for co-existing pathogens, reinforcing the need for standardized preparation protocols and further investigation into optimal viral dosing for effective immunization. Full article