This study compared the growth performance, rumen fermentation patterns, and microbial community structure between three-way crossbred (Poll Dorset × Southdown × Hu, TNH) and purebred Hu (HH) sheep. A total of 75 male lambs (HH: n = 35; TNH: n = 40) with comparable birth dates were selected. After a 56-day suckling period, all lambs underwent an 84-day standardized fattening protocol with a uniform diet. Growth indicators were monitored, and rumen fluid was collected using an oral ruminal sampler for the determination of volatile fatty acids (VFAs) and 16S rRNA sequencing. TNH sheep demonstrated significantly superior growth performance, including higher body weight (BW), average daily gain (ADG) and Kleiber ratio (KR). TNH sheep exhibited an acetate-type pattern with higher acetate, butyrate, total VFAs, and acetate/propionate ratio (A/P), whereas HH sheep showed a propionate-type fermentation pattern with higher branched-chain VFAs. Bacteroidetes and Firmicutes were the dominant core microbiota in both the TNH and HH groups. However, the higher species richness and diversity in the TNH sheep was observed, and harbored a distinct community structure featuring Prevotella as the predominant genus. Shuttleworthia, Olsenella, and Syntrophococcus were unique to HH sheep, whereas Prevotellaceae_UCG_003 was specifically enriched in the TNH sheep. Metabolic pathways, carbon metabolism, and glycolysis/gluconeogenesis were predominantly enriched in TNH sheep, whereas biosynthesis of amino acids and ABC transporters were more abundant in HH sheep. These findings demonstrate that crossbreeding can optimize host metabolism and production performance by modulating rumen VFA dynamics and microbiota structure.

Dengue virus serotype 2 (DENV-2) is a major driver of severe dengue outbreaks globally, yet its evolutionary history and transmission dynamics across Africa remain incompletely understood. To clarify its origins, introduction pathways, and patterns of regional spread, we compiled all available full-length DENV-2 genomes up to 16 July 2025, prioritizing African sequences while retaining representative global references. After quality filtering and down-sampling, a final dataset of 414 genomes was analyzed. Sequences were screened for recombination using RDP4 and analyzed with maximum-likelihood phylogenetics in IQ-TREE with 1,000 ultrafast bootstrap replicates. Temporal dynamics were inferred using time-scaled phylogenies reconstructed in BEAST under a strict molecular clock and exponential-growth coalescent prior, while viral dispersal was examined through discrete-trait phylogeographic analyses with Bayesian stochastic search variable selection. In parallel, single-nucleotide polymorphisms in the prM, E, and NS1 genes were characterized, and significant sites were mapped onto corresponding protein structures. Between 1964 and 2024, African DENV-2 diversity was shaped primarily by repeated introductions of the Cosmopolitan genotype, giving rise to geographically structured lineages. Lineage IIB, likely originating from La Réunion in the late 1970s, subsequently disseminated across West, Central, and East Africa, with Burkina Faso emerging as a key regional hub. Lineage IIA, also linked to La Réunion, diverged into two sub-lineages that expanded through East and Central Africa and extended into South Asia. In contrast, lineages IID and IIF were detected only sporadically, with limited geographic representation and no evidence of sustained regional expansion. Despite substantial genetic variation across prM, E, and NS1, relatively few significant non-synonymous mutations were identified, and critical N-glycosylation motifs remained conserved. This pattern suggests that epidemiological connectivity and repeated viral introductions, rather than sustained positive selection at major functional loci, are the dominant forces shaping DENV-2 spread in Africa. Overall, our findings highlight the central role of recurrent introductions and regional transmission networks in structuring DENV-2 circulation across the continent and emphasize the need for strengthened genomic surveillance to track lineage emergence and improve evidence-based dengue prevention and control strategies.

Plant growth, soil health, and crop productivity with nutritional quality can be significantly enhanced by employing microbial consortia that incorporate diverse microorganisms with complementary functions. Plants produce various types of secondary metabolites such as terpenoids, alkaloids, phenolics, essential oils, and other metabolites through various cellular mechanisms, which are often stimulated by microbial interactions. These metabolites exert beneficial effects on plants and perform multiple roles in agriculture, contributing significantly to growth and economy. This review summarizes microbial consortia-mediated enhancement of plant health and their intricate interactions with host plants. Beneficial microbes of a consortium trigger complex signaling cascades leading to a dynamic regulatory strategy through which plants enhance their secondary metabolite synthesis. Secondary messengers and hormonal cross-talk further integrate the signal to transcription factors, which play a central role in activating or repressing the key genes of the metabolic pathways. Thus, the interplay of microbial signal, secondary messengers, hormonal cross-talk, and key metabolite genes forms the basis of plant secondary metabolite biosynthesis. In addition, recent advances in systems microbiology, including metagenomics, metatranscriptomics, and metabolomics, have enabled a holistic understanding of microbial community dynamics and their collective role in regulating secondary metabolism.

Vector-borne and other zoonotic RNA viruses provide a significant and growing threat to global health, especially in areas where climate change, urbanization, and population growth facilitate the proliferation of arthropod vectors. This review offers an extensive examination of the biology, epidemiology, and pathogenesis of numerous important viruses, including dengue, Zika, chikungunya, yellow fever, Japanese encephalitis, Crimean-Congo hemorrhagic fever, Nipah, Ebola, and hantaviruses. We underscore how environmental and social factors, particularly increasing temperatures, modified precipitation patterns, and accelerated urbanization, transform vector habitats and spillover dynamics. The article further analyzes host-virus and virus-vector interactions, highlighting mechanisms of immune evasion, neurotropism, and vascular disease. Computational and machine learning models are examined as novel instruments for forecasting outbreaks and developing early warning systems. Finally, a summary of present and prospective control options is provided, covering integrated vector management, Wolbachia-based biological control, vaccinations, and antiviral immunotherapies.

Colorectal cancer (CRC) remains a leading cause of cancer-related death worldwide. Primary sclerosing cholangitis (PSC) increases the risk of CRC, but the biological link is not well understood. Changes in the gut-liver axis, including gut microbiota and microbial metabolites, may contribute to CRC development in PSC. To explore pathways connecting PSC with CRC through gut microbiota signals, and to identify candidate host genes for follow-up analyses. Mendelian randomization (MR) was used to estimate the causal effect of genetically predicted PSC on CRC risk and to test associations between PSC liability and gut microbial traits. Mediation analyses were performed to assess whether specific microbial features may contribute to the PSC-CRC association. Single-cell RNA sequencing (scRNA-seq) data from PSC and CRC were integrated to screen candidate host genes, which were further evaluated using survival analyses and immunohistochemistry. MR supported a causal association between PSC and higher CRC risk (OR = 1.172, 95% CI: 1.033-1.329, P = 0.014). Genetically predicted PSC was associated with changes in gut microbial composition, with the family Lachnospiraceae emerging as a notable feature. Within these signals, the genus Lachnospiraceae FCS020 group was associated with CRC risk (OR = 1.237, 95% CI: 1.004-1.523, P = 0.045) and partially mediated the PSC-CRC association (mediating proportion 13.7%, 95% CI: 1.0%-68.1%; indirect effect 0.021, 95% CI: 0.000-0.054). In scRNA-seq analyses, PCBP1 emerged as a shared signal across PSC and CRC programs and was linked to butyrate related gene signatures. Higher PCBP1 expression was associated with poorer overall survival (OS) and recurrence-free survival (RFS) in CRC, and PCBP1 showed a positive association with PD-L1 by immunohistochemistry in an exploratory analysis. These findings provide supportive evidence that gut microbiota changes and host gene signals are linked to the PSC-CRC association. Lachnospiraceae related microbial features and PCBP1 warrant further validation. Studies in independent cohorts and mechanistic models will be important to clarify functional specificity and clinical utility.

Aedes albopictus is the primary vector of major arboviral diseases such as dengue fever, chikungunya fever, and Zika virus disease, and its control is highly dependent on chemical insecticides. However, the long-term use of pyrethroid insecticides has led to the development of insecticide resistance in Ae. albopictus, which severely undermines the efficacy of vector control programs. Ae. albopictus populations were collected from five sites in Guangdong and Hainan provinces, China. Beta-cypermethrin resistance levels were determined via bioassays, with resistance ratios at the median lethal concentration (RR50) calculated. Target-site resistance was evaluated via kdr mutation detection in the voltage-gated sodium channel (VGSC) gene. Transcriptome sequencing identified differentially expressed genes (DEGs), and 16S rRNA sequencing characterized gut microbiome alterations. Correlation analysis and Cedecea neteri dietary supplementation assays verified the role of gut microbiota in resistance. The results showed that all four populations (except the CP population) exhibited varying degrees of resistance to beta-cypermethrin, with resistance ratios at the median lethal concentration (RR50) ranging from 2.84 to 29.18. Detection of kdr mutations revealed three mutations (F1534C, F1534L, F1534S) at codon 1534 of the voltage-gated sodium channel (VGSC) gene in all field populations, with mutation frequencies ranging from 49.4% to 100.0%, and a low-frequency V1016G mutation at codon 1016. Transcriptome analysis identified a total of 2,566 commonly upregulated genes and 994 commonly downregulated genes across the resistant populations. Gut microbiome analysis revealed a significant alteration in the intestinal microbial community structure of resistant populations; specifically, the relative abundance of the genus Cedecea differed significantly between resistant and susceptible populations and correlated strongly with the expression of most differentially expressed genes. Furthermore, dietary supplementation with Cedecea neteri significantly increased the survival rate of Ae. albopictus exposed to β-cypermethrin (73.86% vs 40.00%; P < 0.0001). From the perspectives of target-site mutations, gene expression regulation, and gut microbe interactions, this study providing a foundation for further studies on resistance mechanisms in Ae. albopictus, thereby providing a theoretical foundation for further dissection of resistance mechanisms and optimization of vector control strategies.

Single-stranded DNA (ssDNA) is an essential intermediate of genome duplication but can also arise in the genome, including at highly transcribed loci. Although the origin recognition complex (ORC), a eukaryotic replication initiator, has been reported to bind ssDNA, how interactions with ssDNA-exposing regions are regulated without compromising origin specificity and genome stability remains poorly understood. Here, we characterize the ssDNA-binding properties of budding yeast ORC and the role of an initiator-specific motif (ISM)-related region within the AAA+ domain of Orc1. In vitro, ORC binds ssDNA (except for poly dA) at affinities comparable to those of replication protein A (RPA) under identical binding conditions, suggesting base composition-dependent modulation of ORC-ssDNA binding. Genome-wide analyses show partial overlap between ORC- and RPA-enriched peaks, consistent with ORC binding at a subset of ssDNA-forming loci in vivo. Mutations of the Orc1 ISM-related region increase ORC binding to ssDNA and promote higher-order ORC-ssDNA complex formation in vitro, indicating that this region normally limits ORC-ssDNA binding. In contrast, these mutations impair origin-specific ORC binding through mechanisms involving the essential A-element-proximal region. Genetic and chromatin-based analyses further reveal that enhanced ssDNA binding correlates with reduced origin binding in vivo, indicating a redistribution of ORC from replication origins to ssDNA-forming loci. Despite conservation of the ISM across domains of life, these results suggest that eukaryotic ORC has functionally diverged such that origin binding is coupled to the repression of ssDNA binding through the Orc1 ISM-related region, thereby safeguarding faithful origin selection.

Soybean root rot, a devastating soil-borne disease, severely limits global soybean yield and quality. Traditional control measures cause environmental pollution and have regional limitations. Root-associated microbiomes are vital for plant health, but most studies use relative abundance sequencing that distorts actual microbial quantities. This study used absolute quantitative high-throughput sequencing to clarify soil chemical properties and bacterial community assembly in healthy and diseased soybeans, laying a theoretical foundation for microbiome-based root rot management. Samples were collected from bulk soil, and the endosphere, rhizoplane, rhizosphere of healthy and diseased soybeans in black soil fields of Heilongjiang. Soil chemical properties, including pH, soil organic carbon (SOC), total nitrogen (TN), available nitrogen (AN), available phosphorus (AP), and available potassium (AK), were determined. Absolute quantitative sequencing of bacterial 16S rRNA V4-V5 region was performed, combined with qPCR for absolute abundance calibration. Bioinformatics analyses included α/β diversity, co-occurrence network, community assembly (βNTI & RCbray), random forest and correlation analysis to identify key taxa and their relationships with environmental factors. Root rot significantly reduced rhizosphere SOC (by 29.13%), TN (8.57%), AN (24.18%), AP (18.86%), while increased AK (12.82%) and pH. However, the contents of certain bacterial taxa at the genus levels showed significant differences in both absolute and relative abundances. The bacterial co-occurrence network indicate that the interaction in the healthy soybean (H) group was more complex than that in the diseased soybean (S) group. Specifically, 1 module hub and 21 connectors were identified in the H group, whereas 55 connectors were detected in the S group. Community assembly in both the H and S groups was governed by deterministic processes, with homogeneous selection primarily observed in the S group. Random forest and correlation heatmap analyses revealed ASV115 (Candidatus Koribacter) in H group was positively correlated with SOC, pH and AN; ASV16 (Streptomyces), ASV42 (Agrobacterium) and ASV46 (Mesorhizobium) were keystones in S group. Root rot destroyed rhizosphere nutrient balance and reshaped bacterial community structure, reducing network complexity. Absolute quantification effectively compensated for the defects of relative abundance, accurately reflecting community changes. These findings clarify the microecological mechanism of soybean root rot, supporting the development of biocontrol strategies targeting rhizosphere microbiome for sustainable soybean production.