Search Results (658)

We report a case of catheter-associated bloodstream infection caused by a putative novel species in the genus Erwinia, identified using whole-genome sequencing (WGS). A female adolescent receiving long-term home parenteral nutrition via a central venous catheter (CVC) presented with a fever. Gram-negative rods were isolated from two CVC-derived blood culture sets, while peripheral cultures remained negative. Conventional identification methods, including VITEK 2, Phoenix M50, MALDI-TOF MS, and 16S rRNA and rpoB gene sequencing, failed to achieve species-level identification. WGS was performed on the isolate using Illumina MiSeq. Genomic analysis revealed a genome size of 5.39 Mb with 56.8% GC content and high assembly completeness. The highest average nucleotide identity (ANI) was 90.3% with Pantoea coffeiphila, and ≤85% with known Erwinia species, suggesting that it represents a distinct taxon. Phylogenetic analyses placed the isolate within the Erwinia clade but separate from any known species. Antimicrobial susceptibility testing showed broad susceptibility. This case highlights the utility of WGS for the identification of rare or novel organisms not captured by conventional methods and expands the clinical spectrum of Erwinia species. While the criteria for species delineation were met, the phenotypic characterization remains insufficient to formally propose a new species. Full article

Soil bacteria drive biogeochemical cycles and influence disease suppression, playing pivotal roles in sustainable agriculture. Using Illumina MiSeq sequencing, we assessed how six ridge-furrow film mulching patterns affect soil bacterial diversity in a continuous potato system. The Shannon index showed significantly higher diversity in fully mulched treatments (T2–T3) versus controls (CK), suggesting mulching enhances microbial community richness. This result suggests that complete mulching combined with ridge planting (T2) may significantly enhance bacterial proliferation in soil. The bacterial communities were predominantly composed of Acidobacteria, Pseudomonadota, Bacteroidota, Chloroflexota, and Planctomycetota. Among these, Acidobacteria showed the highest abundance, with ridge planting patterns favoring greater Acidobacteria richness compared to furrow planting. In contrast, Pseudomonadota exhibited higher abundance under half-mulching conditions than under complete mulching. At class level, Acidobacteria and Proteobacteria emerged as the most abundant groups, with Proteobacteria constituting 22.6–35.7% of total microbial populations. Notably, Proteobacteria demonstrated particular dominance under the complete mulching with ridge planting pattern (T2). At the genus level, Subgroup_6_norank represented the most dominant taxon among the 439 identified bacterial genera, accounting for 14.0–20.2% of communities across all treatments, with half-mulching ridge planting (T4) showing the highest relative abundance. Our findings demonstrate that different ridge-furrow film mulching patterns significantly influence soil microbial diversity. While traditional non-mulched (CK) and mulched flat plots (T1) exhibited similar impacts on bacterial community structure, other treatments displayed distinct taxonomic profiles. Complete mulching patterns, particularly ridge planting (T2), appear most conducive to microbial development, suggesting their potential to enhance soil biogeochemical cycling in continuous cropping systems. These results provide valuable insights for optimizing mulching practices to improve soil health in agricultural ecosystems. Full article

Bacterial symbionts play an important role in insect survival by contributing to key metabolic and defensive functions. While stingless bees are known to harbor diverse microbial communities, their core bacterial symbionts remain poorly characterized. In this study, we analyzed the gut microbiota of sixteen stingless bee species collected from different regions of Mexico using 16S rRNA gene sequencing on the Illumina^® MiSeq™ platform. Our results revealed that Proteobacteria, Firmicutes, and Actinobacteria are the most abundant bacterial phyla across species. Among the dominant genera, lactic acid bacteria, such as Lactobacillus spp., Bifidobacterium, and Fructobacillus spp., were the most prevalent. These bacteria are responsible for developing biochemical functions in metabolic processes like lactic fermentation and the biotransformation of complex organic compounds into molecules that are more easily assimilated by bees. This study offers a novel perspective on the diversity and predicted composition of gut microbiota in Mexican stingless bees. By highlighting differences in microbial communities among species with different feeding habits, our results emphasize the importance of preserving microbial biodiversity in these pollinators. Full article

This research aimed to study changes in the diversity of the soil bacterial community in a jujube forest with different years of abandonment. To this end, we took the mountain jujube forest with different abandoned years (1 a, 3 a, 6a and 20 a) in the Qijiashan jujube experimental demonstration base in Yanchuan County as the research object; we used Illumina Miseq high-throughput sequencing technology to analyze the changes in the soil bacterial community structure and reveal the key environmental drivers of bacterial community variation in the abandoned jujube forest in the study area. The results showed the following findings: (1) Phylum Actinomycetota (34%), Proteobacteria (29%), and Acidobacteriota (13%) were the dominant phyla of the soil bacterial community in the abandoned jujube forest. (2) Abandonment altered the composition of soil bacteria at the OTU level in jujube plantations. (3) There are differences in the soil bacterial community structure across different periods of abandonment in the jujube forest. (4) Soil water content is the main factor affecting the bacterial community structure of the abandoned jujube forest. There are differences in the soil water content of abandoned woodlands, which affects the community structure of soil microorganisms. Full article

Potato early dying disease complex (PED) leads to premature senescence and rapid decline in potato plants. Unlike potato wilt caused solely by Verticillium species, PED symptoms are more severe due to the synergistic effects of multiple pathogens, including root-lesion nematodes, fungi such as Colletotrichum and Fusarium, and soft-rot bacteria. To investigate the microbiome responsible for PED, soil and stem samples from healthy-looking and symptomatic plants were analyzed using amplicon-targeted next-generation sequencing (Illumina MiSeq and PacBio technologies). Samples were collected from four locations in New Brunswick, Canada from fields previously rotated with barley or oat. Comparative analysis of the bacterial, fungal, and eukaryotic diversity in soil samples showed minimal differences, with only bacterial alpha diversity influenced by the plant health status. Verticillium dahliae was abundant in all soil samples, and its abundance was significantly higher in the stems of diseased plants. Additional fungal species implicated in PED, including Plectosphaerella cucumerina, Colletotrichum coccodes, Botrytis sp., and Alternaria alternata, were also identified in the stems. This study highlights the complex, plant-associated microbial interactions underlying PED and provides a foundation for microbiome-informed disease management strategies. Full article

Excessive fertilizer input and low output are currently problems for peach production in China. Sugar alcohols such as sorbitol and mannitol represent promising eco-friendly fertilization strategies to improve fruit quality and optimize nutrient management. Our research explored the effect of sorbitol and mannitol on the rhizosphere environment and peach growth from the rhizosphere micro-ecology perspective. Potted peach seedlings were used as materials. Without adding or adding different sorbitol and mannitol concentration gradients (100, 200, 400) combined with potassium dihydrogen phosphate (KH₂PO₄), the physicochemical properties of rhizosphere soil, leaf nutrition, photosynthetic and growth index were determined, and the rhizosphere bacterial community was analyzed via Illumina Miseq high-throughput sequencing. Both sorbitol and mannitol altered the rhizosphere environment, effectively improved leaf photosynthesis, and promoted peach seedling growth; particularly, M100 had optimal affection. Sorbitol and mannitol altered the bacterial structure and reduced bacterial diversity, which observably correlated with soil organic matter and available potassium. For the rhizosphere bacterial composition, sorbitol and mannitol increased specific bacterial OTUs and induced changes in bacterial composition, among which chemoheterotrophic and nitrogen-transforming bacteria increased with the addition of sorbitol and mannitol. Association network analysis and a structural equation model showed that S100 and M100 mainly enriched Vicinamibacteraceae to regulate peach seedling growth. Overall, low-concentration sorbitol and mannitol showed the best effect in peach seedling growth through regulating the rhizosphere environment. Full article

Background: Antimicrobial resistance (AMR) is a critical global health threat, with AMR Salmonella enterica serovar Typhimurium strains being a major foodborne pathogen. Integrons, a type of mobile genetic element, capture and transfer resistance genes, thereby playing a role in the spread of AMR. Objectives: This study aimed to characterize the locations of integrons carrying AMR genes within the whole genomes of 32 Salmonella Typhimurium isolates collected from dairy cattle by two U.S. Veterinary Diagnostic Laboratories between 2009 and 2012. Methods: Class I integrons were sequenced from PCR-amplified products. DNA was extracted, quantified, barcoded, and sequenced on the Illumina MiSeq platform. Whole genome sequences were trimmed and assembled using the SPAdes assembler in Geneious Prime^®, and plasmids were identified with the PlasmidFinder pipeline in Linux. Integron locations were determined by aligning their sequences with whole genome contigs and plasmids, while AMR genes were identified through BLAST with the MEGARes 3.0 database and confirmed by alignment with isolate, plasmid, and integron sequences. Statistical analysis was applied to compare the proportions of isolates harboring integrons on their chromosome versus plasmids and also to examine the associations between integron presence and AMR gene presence. Results: Seven plasmid types were identified from all isolates: IncFII(S) (n = 14), IncFIB(S) (n = 13), IncC (n = 7), Inc1-I(Alpha) (n = 3), and ColpVC, Col(pAHAD28), and Col8282 (1 isolate each). Of the 32 isolates, 16 (50%) carried at least one size of integron. Twelve of them carried both 1000 and 1200 bp; 3 carried only 1000 bp and 1 carried 1800 bp integrons. Of the 15 isolates that carried 1000 bp integron, 12 harbored it on IncFIB(S) plasmids, 2 on IncC plasmids, and 1 on the chromosome. The 1200 bp integrons from all 12 isolates were located on chromosomes. There were significant positive associations between the presence of integrons and the presence of several AMR genes including sul1, aadA2, blaCARB-2, qacEdelta1, tet(G), and floR (p < 0.05). AMR genes were located as follows: aadA2 on IncFIB(S) and IncC plasmids; bla_CMY-2 on IncC plasmid; qacEdelta1 on IncFIB(S), IncC, and chromosome; bla_CARB-2, floR, tet(A) and tet(G) on the chromosome. Conclusions: The findings highlight the genomic and plasmid complexity of Salmonella Typhimurium which is impacted by the presence and location of integrons, and this study provides genomic insights that can inform efforts to enhance food safety and protect both animal and public health. Full article

Background: Antimicrobial resistance (AMR) poses significant risks to human and animal health, while the environment can contribute to its spread. National AMR surveillance programs are pivotal for assessing AMR prevalence, trends, and intervention outcomes; however, integrating advanced surveillance tools can be difficult. This pilot study, conducted by FAO ECTAD Indonesia and DGLAHS, the Indonesian Ministry of Agriculture, evaluated the costs and benefits of integrating the Nanopore MinION, Illumina MiSeq, and Sensititre system into a culture-based slaughterhouse–river surveillance system. Methods: Water samples were collected from six chicken slaughterhouses and adjacent rivers (pre- and post-treatment effluent, upstream, and downstream). Culture-based ESBL and general E. coli concentrations were estimated via the WHO Tricycle Protocol, while isolates (n = 42) were sequenced (MinION, MiSeq) and antimicrobial susceptibility testing conducted (Sensititre). Results: The Tricycle Protocol results provided estimates of effluent and river concentrations of ESBL and general E. coli identifying ESBL-to-general E. coli ratios of 13.8% and 6.2%, respectively. Compared to hybrid sequencing assemblies, MinION had a higher concordance than MiSeq for ARG identification (98%), virulence genes (96%), and locations for both (predominately plasmids). Furthermore, MinION concordance with Sensititre AST was 91%. Conclusions: Cost–benefit comparisons suggest sequencing can complement culture-based methods but is dependent on the value placed on the additional information gained. Full article

Outbreaks of infectious diseases contribute significantly to morbidity and mortality in resource-limited settings, yet the capacity to identify their etiology remains limited. We aimed to characterize microbes and antimicrobial resistance (AMR) genes in Tanzanian children affected by an acute febrile illness (AFI) outbreak using metagenomic next-generation sequencing (mNGS). A cross-sectional study was conducted on archived blood samples from children who presented with AFI between 2018 and 2019. Total nucleic acids were extracted from 200 µL of blood, and complementary DNA (cDNA), along with enriched pathogenic DNA, was sequenced using the Illumina MiSeq platform. mNGS data were analyzed using CZ-ID Illumina mNGS bioinformatics pipeline v7.0. Results were obtained from 25 participants (mean age: 11.6 years; SD ± 5), of whom 36% had a moderate to high-grade fever. The following five potential microbial causes of AFI were identified: Escherichia coli (n = 19), Paraclostridium bifermentans (n = 2), Pegivirus C (n = 2), Shigella flexneri (n = 1) and Pseudomonas fluorescens (n = 1), with E. coli being the most prevalent. Twelve AMR genes were detected, including mdtC, acrF, mdtF, and emrB. E. coli harbored most of the AMR genes previously associated with resistance to commonly used antibiotics. mNGS offers a promising complementary approach to conventional diagnostics for identifying pathogens and AMR profiles in vulnerable populations. Full article

The intestinal microbiota is vital for host immunity and metabolism, and its changes are associated with the development stage of hosts. However, little is known regarding how growth and development of anurans affect the diversity of their microbiota, which has a complex life cycle. The Tibetan toad (Bufo tibetanus) is a wild population in the high-altitude area of southwest China, which has special adaptability to the environment. Here, the microbial community of the Tibetan toad at six developmental stages (from the tadpole at Gosner stage 18 to the 8-year-old adult) was assessed using high-throughput 16S rRNA sequencing. The alpha diversity index analysis showed that the Chao, Ace, and Shannon indices were highest at Gosner stage 32 and decreased as development progressed, and their alpha diversity remained unchanged over time in adult stages. Beta diversity revealed that the gut microbiota structure differed significantly from Gosner stages 18 to 31, and it became similar to adult toads from Gosner stages 45 to 46 and in juvenile groups. At the phylum level, Firmicutes, Proteobacteria, and Actinobacteria were dominant phyla in tadpoles and adults. The relative abundance of Firmicutes and Proteobacteria in the adult group was significantly higher and lower than that of tadpoles, respectively. The linear discriminant analysis effect size (LEfSe) analysis identified seven phyla exhibiting significant differences during life stages: Verrucomicrobiota, Bacteroidota, and Proteobacteria (Gosner 18 to 31), Cyanobateria and Chloroflexi (Gosner 32 to 41), Actinobacteriota (Gosner 45 to 46), Desulfobacterota (juvenile group), and Firmicutes (adult group). A pathway enrichment analysis revealed that the metabolism and biosynthesis of secondary metabolites were significantly enriched across all developmental stages. This research unveiled variations in the intestinal microbiota composition during development in anurans. Factors such as developmental stage, habitat type and feeding habit jointly affected the gut microbial diversity and community composition in the Tibetan toad. The findings of this study can provide information for understanding the influence of historical developments on the intestinal microbiota and provide protection information for anurans. Full article

We have used a rapid, portable assay (Bacterisk) to determine the bacterial water quality along several inland waters in SW England. Water samples were compared by a conventional membrane filter and culture methods for faecal indicator bacteria (FIB; E. coli and enterococci) and endotoxin measurement by Bacterisk. The Bacterisk data, measured in near-real-time, correlate well with both E. coli and enterococci, but also allow the presence of potential pathogens of a non-faecal origin to be detected. The sensitivity was calculated to be 92.96% with a specificity of 46.3% for E. coli with an expanded uncertainty of 22.07% and an Endotoxin Risk detection limit of 25 units. The presence of Bacterisk detectable non-faecal pathogenic bacteria in the water samples was successfully confirmed by Illumina MiSeq sequencing followed by target species-specific qPCR. Sequencing showed the presence of pathogens including Pseudomonas aeruginosa, Salmonella typhi, Acinetobacter baumannii, Shigella spp., and Legionella spp. as well as antimicrobial resistance genes. Furthermore, the portable Bacterisk assay was able to acquire data on the water quality from different locations and at different time points, providing a comprehensive surveillance tool that challenges the time to results by conventional methods (minutes instead of days), yielding compatible results. Full article

Soil fungi serve as key mediators of belowground ecological processes; however, the altitudinal distribution patterns and their driving mechanisms of soil fungal communities in alpine shrubland ecosystems remain poorly understood. In this study, soil samples were collected from Potentilla fruticosa shrubs at different altitudes, and their physical and chemical properties were determined. Illumina MiSeq sequencing technology was used to study the characteristics of soil fungal communities at different altitudes (3400, 3700, 4000, and 4300 m), and the driving factors affecting the composition of soil fungal communities were found through variance analysis and redundancy analysis. With the increase in altitude, species diversity decreased while total phosphorus and available phosphorus increased. Compared with 3400 m, the diversity index (Sobs, Chao1, and ACE index) of the soil fungal community at 4000 m is the highest, and that at 4300 m is the lowest. NMDS analysis showed that there were significant differences among soil fungal community structures at different altitudes. Redundancy analysis (RDA) indicated that available potassium, available phosphorus, and the Shannon–Wiener diversity index were the primary factors influencing the variation in soil fungal communities along the elevation gradient. Furthermore, the impact of soil physical and chemical properties on soil fungal communities was found to be more pronounced than that of plant characteristics. Network analysis shows that the network complexity is the highest at 4300 m above sea level. These studies provide a new perspective and basis for understanding the distribution pattern of soil fungi in the rhizosphere Potentilla fruticosa in the eastern Qinghai–Tibet Plateau. Full article

Ecuador, a tropical country with frequent dengue outbreaks, including a surge from 16,017 cases in 2022 to 61,329 in 2024, was the focus of this study. The study was conducted in Borbon, a semi-urban rural town in the Esmeraldas province. Genomic analysis, alongside entomo-virological surveillance, provides valuable insights into DENV-2 genotypes. Five pools of female Aedes aegypti mosquitoes from Borbon tested positive for DENV serotype 2 through RT-qPCR. One positive pool (CT = 16.13) was sequenced using Illumina MiSeq, and genotyping was conducted via the Dengue Typing Tool and Maximum Likelihood phylogenetic tree. The genotype assigned was III Southern Asian-American. Comparison with other genomes revealed genetic similarity to a human dengue genome sequenced in 2021, also from Esmeraldas, clustering with genomes reported across the Americas, particularly from Colombia and Venezuela. This study enhances our understanding of dengue virus epidemiology in rural areas, emphasizing the critical role of clinical case surveillance and vector monitoring in guiding evidence-based interventions. Full article

In this study, 172 Single-Nucleotide Polymorphisms (SNPs) (94 identity-informative SNPs, 56 ancestry-informative SNPs, and 22 phenotypic-informative SNPs) included in the ForenSeq™ DNA Signature Prep kit/DNA Primer Mix B (Verogen) were used for genotyping DNA samples from a population of twenty-one unrelated subjects, native to Northeast Italy. SNP sequencing was performed with the MiSeq FGx™ Forensic Genomics System (Illumina-Verogen), and data were analyzed using the Universal Analysis Software (UAS) v1.2. Raw data underwent further examination with STRait Razor v3 (SRv3) to compare the target SNPs’ genotype calls made with UAS and to identify the presence of microhaplotypes (MHs) due to SNPs associated with the same target SNP’s amplicon. The allele (haplotype) frequencies, Hardy–Weinberg equilibrium, linkage disequilibrium, number of effective alleles (A_e), and relevant forensic statistic parameters were calculated. Among the 172 SNPs evaluated, 45 unique microhaplotypes were found, comprising a novel sequence variant never previously described. The presence of MHs resulted in an 8.00% rise in the typologies of unique sequences, leading to changes in A_e. Notably, for 12 out of the 94 iiSNPs, the values of A_e exceeded 2.00, which is generally associated with a higher expected heterozygosity and increased power of discrimination. Full article

Pinus tabuliformis is a well-recognized woody mycorrhizae host plant growing in North China. EM fungi contribute to the host health and the stability of the forest ecosystem. However, ectomycorrhiae (EM) fungal community associated with this species is less documented. In this study, we examined EM fungal diversity and composition of P. tabuliformis from three sites in Inner Mongolia, China by using Illumina MiSeq sequencing on the rDNA ITS2 region. Our results showed that a total of 105 EM fungal operational taxonomic units (OTUs) were identified from 15 composite root samples, and the dominant lineages were /suillus-rhizopogon, /tomentella-thelephora, /tricholoma, /amphinema-tylospora, /wilcoxina, /inocybe, and /Sebacina. A high proportion of unique EM fungal OTUs (33, 31.4% of total OTUs) were detected, and some abundant OTUs preferred to exist in specific sites. The composition of EM fungal communities was significantly different among the sites, with soil, climatic, and spatial variables being related to the community variations. The EM fungal community assembly was mainly driven by environmental factors in deterministic processes. These findings suggest that this endemic Pinaceae species in China also harbored a rich and distinctive EM fungal community and deterministic processes played more important roles than stochastic in shaping the symbiotic fungal community. Our study improves our understanding of EM fungal diversity and community structure from the perspective of a single host plant that has not been investigated exclusively before. Full article