Search Results (1,344)

In the Iberian Peninsula, the rapid expansion of the invasive species Alburnus alburnus (bleak) has intensified its contact with several endemic cyprinid species, raising concerns about hybridization, introgression, and the loss of genetic diversity. Despite increasing evidence of hybridization, data remain limited for many Iberian River basins, where endemic species persist in fragmented and vulnerable habitats. The aim of this study is to assess the extent and spatial distribution of hybridization between the bleak and the following native cyprinid species, Anaecypris hispanica (jarabugo), Iberochondrostoma lemmingii (pardilla), Pseudochondrostoma willkommii (Guadiana nase), and Squalius alburnoides (calandino), across several rivers within the Guadalquivir Basin. To this end, Sanger sequencing will be performed on the mitochondrial cytochrome b gene (maternal lineage; approximately 1000 base pairs (bp)) and the nuclear β-actin gene (paternal lineage; approximately 950 bp) from individuals of all endemic species and the bleak. Parental species and putative hybrids were initially identified in the field using diagnostic morphological and meristic characters, including number of rays in the anal fin morphology, mouth position, and the number of lateral line scales. Molecular analyses will include haplotype network reconstruction and phylogenetic tree inference to evaluate relationships among individuals from different species and to assess lineage divergence. The results will allow us to: (1) detect hybrids between the bleak and endemic cyprinids, (2) identify hybridization events among endemic cyprinid species, and (3) evaluate the correspondence between diagnostic morphological and meristic traits and the molecular identification of hybrid individuals. Overall, these findings will provide key information for the conservation management of endangered Iberian freshwater fishes in the context of invasive species expansion and global change. Full article

Background: Benzimidazole (BZ) resistance in the gastrointestinal nematode Haemonchus contortus is a major constraint to sheep production worldwide. However, data on the prevalence and molecular mechanisms of resistance in Yili Prefecture, Xinjiang—a key livestock region in Northwestern China—remain limited. This study aimed to determine the frequency of BZ resistance-associated single nucleotide polymorphisms (SNPs) in H. contortus populations from Zhaosu and Tekesi counties. Methods: Adult male worms (n = 150) were collected from naturally infected sheep at local abattoirs. Species identity was confirmed morphologically by sequencing the internal transcribed spacer 2 (ITS-2) region. A 385 bp fragment of the isotype-1 β-tubulin gene was amplified and sequenced to detect SNPs at codons 167 (F167Y), 198 (E198A), and 200 (F200Y). Results: The F167Y mutation was absent in all individuals. In contrast, the E198A mutation occurred at exceptionally high frequencies, with resistant allele frequencies (RAF) of 64.7% in Zhaosu and 52.7% in Tekesi. The F200Y mutation showed clear geographical variation: it remained low in Zhaosu (RAF = 9.3%) but was substantially higher in Tekesi (RAF = 33.3%). Haplotype analysis revealed that resistance in Zhaosu was driven primarily by the E198A mutation, whereas the Tekesi population exhibited complex patterns of co-selection of both E198A and F200Y, with a high proportion of double-heterozygous individuals (29.3%). Conclusions: This study provides comprehensive molecular evidence of severe BZ resistance in H. contortus populations from Zhaosu and Tekesi counties, Yili Prefecture. The marked predominance of the E198A mutation, together with the emergence of multi-locus resistance in Tekesi, indicates a rapid escalation of resistance beyond historical levels. These findings suggest that benzimidazoles are likely ineffective in this region and highlight the urgent need to revise local parasite control strategies. Full article

Surface-enhanced Raman scattering (SERS) offers a powerful route for biomolecular detection because it combines molecular specificity with high sensitivity, rapid optical readout, and multiplexing capability. In real biological samples, however, analytical performance is rarely determined by signal enhancement alone. Biofluids such as serum, plasma, saliva, urine, and interstitial fluid contain complex biomolecular mixtures that interfere with target capture, spectral response, and data interpretation. A practical SERS biosensor must therefore localize targets, stabilize spectral responses, tolerate matrix-induced variation, and convert complex spectra into reliable analytical information. This review discusses recent progress in SERS biosensing from an integrated system perspective, with particular focus on artificial intelligence/machine learning (AI/ML)-assisted interpretation. Direct label-free SERS provides chemically transparent readouts but is limited by stochastic adsorption, hotspot heterogeneity, and spectral variation in complex samples. Bio-recognition interfaces improve target localization, while signal-transduction strategies based on nanotags, immunoassays, clustered regularly interspaced short palindromic repeats (CRISPR) systems, nanozymes, and lateral-flow formats decouple molecular recognition from spectral generation. Digital SERS further improves measurement robustness by converting fluctuating intensities into countable, event-based outputs. AI/ML-assisted analysis can support full-spectrum classification, calibration transfer, explainability, and patient-level decision-making. We frame AI/ML-assisted SERS biosensing as an integrated architecture connecting substrate design, interface engineering, signal transduction, digital measurement, and clinical validation. Future progress will depend as much on validation-ready workflows as on plasmonic enhancement itself, especially for systems intended to operate across different samples, instruments, and clinical settings. Full article

Fungal infections pose diagnostic challenges in both human and veterinary medicine, as traditional detection methods such as fungal culture are time-consuming, microscopy is operator-dependent, and molecular detection assays often require specialized instrumentation and trained personnel, which can limit their routine clinical application. This study developed a sandwich immunoassay to detect β-1,3- and β-1,6-glucans, two major components of the fungal cell wall, based on two catalytically inactive glucanase mutants, LamA_E175Q and Neg1_E321Q. The sandwich ELISA exhibited higher detection sensitivity than conventional ITS-based PCR for Saccharomyces cerevisiae and Candida albicans under the conditions of this study. Using pre-coated plates, the sample-processing and detection workflow can be completed in approximately 40 min. It effectively detected a wide range of fungal species, including yeasts (Saccharomyces cerevisiae, Candida albicans) and filamentous fungi such as dermatophytes and non-dermatophyte molds. In a preliminary clinical cohort, the assay identified β-glucan signals in all 21 samples confirmed positive for dermatophytes, while no signal was detected in 20 negative samples, suggesting potential clinical applicability. This dual-enzyme sandwich immunoassay provides a rapid and low-cost complementary tool for broad-spectrum fungal screening, which may help guide further confirmatory diagnostics and timely clinical decision-making. Full article

Per- and polyfluoroalkyl substances (PFASs) are persistent emerging contaminants characterized by high environmental stability and biotoxicity. Ubiquitous detection of these contaminants across aquatic environments poses severe threats to ecosystem stability and human health, while constructed wetlands (CWs) serve as a sustainable low-carbon alternative for the remediation of PFAS-laden wastewater. However, traditional mechanisms such as matrix adsorption, phytoaccumulation, and microbial transformation often suffer from low efficiency, rapid saturation, and incomplete degradation. To overcome the above drawbacks, the arbuscular mycorrhizal fungi (AMF)–plant–microbe synergistic consortium has become a promising remediation candidate, which facilitates PFAS immobilization and biodegradation via symbiotic crosstalk among three components. This paper reviews recent advancements in PFAS remediation within AMF-facilitated systems, examining fundamental synergistic mechanisms, treatment efficiencies, and key influencing factors. We propose several optimization strategies, including substrate modification, operational parameter refinement, and the integration of advanced technologies. Furthermore, we emphasize the necessity of elucidating the molecular pathways governing long-chain PFAS degradation and addressing current bottlenecks in engineering applications. Future research should prioritize molecular interaction level interaction mechanisms, the development of anti-interference systems, and field-scale validation. This review provides a theoretical foundation and technical framework for leveraging AMF–plant–microbe synergism to enhance PFAS removal in CWs. Full article

The hybrid golden pompano “Chenhai No. 1” (CH), generated through distant hybridization [(♀ Trachinotus ovatus × ♂ T. blochii) × ♂ T. ovatus], exhibits significantly enhanced growth performance compared to its parental T. ovatus (TO). To elucidate the molecular mechanisms underlying this rapid growth, we performed integrated transcriptomic and untargeted metabolomic profiling of muscle tissue. Transcriptomic analysis identified 3172 differentially expressed genes (DEGs), with weighted gene co-expression network analysis (WGCNA) highlighting the ‘darkorange2’ module as strongly associated with rapid growth. Key DEGs, including mapk8a, acacb, and pkmb, were upregulated and implicated in energy metabolism, glycolysis, and signal transduction. Metabolomic profiling detected 576 significantly altered metabolites, predominantly enriched in glycolysis, the tricarboxylic acid (TCA) cycle, amino acid metabolism, lipid biosynthesis, and mTOR signaling. Integrated analysis revealed coordinated alterations between core module genes and differential metabolites in interrelated pathways, including correlations between pfkpa/pfkma and glyceraldehyde-3-phosphate, acacb and phosphatidylcholine/phosphatidylethanolamine, and sesn2 and leucine. These findings suggest that the growth advantage of CH arises from the coordinated enhancement of energy metabolism, amino acid sensing, and lipid metabolic remodeling, establishing a synergistic transcription–metabolism regulatory network. This study provides multi-omics insights into the molecular basis of rapid growth in an economically important teleost fish. Full article

(1) Background: Indonesia faces the dual challenge of a high tuberculosis (TB) burden and increasing drug resistance. Conventional molecular diagnostics frequently fail to detect isoniazid resistance and nontuberculous mycobacteria (NTM). This study evaluates a domestic multiplex Open PCR system in Medan, Indonesia. (2) Methods: From July to November 2025, 1569 sputum specimens from suspected TB patients were analysed using the Indigen MTB/NTM/DR-TB Real-time PCR Kit Gen 2. (3) Results: Mycobacterial DNA was detected in 421 specimens (26.8%). Among these, 396 (94.1%) were drug-susceptible TB, while 16 (3.8%) showed resistance, predominantly INH mono-resistance (n = 14; 0.89% of total). Additionally, 9 cases (2.1%) involved NTM or TB-NTM co-infections. Tertiary hospitals showed significantly higher positivity rates (33.5%) than primary care (18.9%; p < 0.001). TB status was significantly associated with male (p = 0.0052) and older age (p = 0.006), whereas resistance profiles and NTM distribution were consistent across all demographic groups (p > 0.80). (4) Conclusions: This study describes the implementation and diagnostic yield of a domestic multiplex Open PCR system in Medan, Indonesia. By bridging diagnostic gaps across a decentralized referral network, this facilitates rapid and targeted therapy. Integrating multiplex domestic innovations into national diagnostic algorithms is essential for achieving Indonesia’s TB elimination targets. Full article

Acute gastroenteritis is a common condition with a wide and often indistinguishable etiology, requiring laboratory support for an accurate diagnosis. Classical diagnostic methods, including stool culture and antigen-based tests, are limited by restricted pathogen coverage and variable sensitivity. In the present study, 548 stool samples from patients with suspected gastroenteritis were tested using the QIAstat-Dx Gastrointestinal Panel 2 and compared with stool culture and rotavirus/adenovirus antigen tests. The molecular panel showed a positivity rate of 50.4%, consistently higher than stool culture (12.6%) and antigen assays (12.3% for rotavirus and 4.4% for adenovirus). The most frequently detected pathogens included enteropathogenic Escherichia coli (15.3%), Campylobacter spp. (12.0%), and enteroaggregative E. coli (10.2%). Agreement between methods was good for bacterial pathogens but low for viral targets. Discordant viral results were often associated with low antigen cut-off index values and multiple pathogen detections by the molecular panel, suggesting potential limitations of antigen-based assays. Overall, the QIAstat-Dx Gastrointestinal Panel 2 improves pathogen detection and provides rapid, comprehensive diagnostic information, while a combined approach with conventional methods may represent the most appropriate strategy for optimizing patient management. Full article

Giardia duodenalis (synonyms G. lamblia and G. intestinalis) and Blastocystis sp. are prevalent zoonotic intestinal protozoans with significant public health and economic importance. Both parasites infect various hosts, including humans and most livestock. G. duodenalis can cause infections ranging from asymptomatic carriage to clinical manifestations such as acute diarrheal disease, growth stunting and malabsorption, whereas Blastocystsi sp. infection is often asymptomatic and its pathogenicity remains debated. Following the rapid expansion of modern equine industries (horse racing, equestrian sports, production, and tourism), concern about the spread of parasitic diseases caused by human–horse contact has gradually increased. In this study, 631 horse fecal specimens were collected from three geographical localities in Shanxi Province, North China. The prevalence and genetic characteristics of the two parasites were determined by amplification of multiple genetic loci (tpi, triosephosphate isomerase; gdh, glutamate dehydrogenase; bg, beta-giardin) for G. duodenalis and the small subunit ribosomal RNA (SSU rRNA) gene for Blastocystis sp. Overall, the prevalence of G. duodenalis and Blastocystis sp. in horses in Shanxi Province was 7.9% and 0.8%, respectively. Sequence analysis identified three assemblages (A, B, and E) of G. duodenalis and two subtypes (ST1 and ST5) of Blastocystis sp. in horses; assemblage B and subtype ST1 were detected more frequently than the other types (subtypes) in this data set. This study is the first to report G. duodenalis and Blastocystis sp. infections in horses in Shanxi Province, and these findings provide baseline molecular epidemiological data for horses in Shanxi Province and support continued hygiene management to reduce potential zoonotic transmission. Full article

Canine dirofilariasis, caused by Dirofilaria immitis and Dirofilaria repens, is an emerging vector-borne disease of increasing veterinary and zoonotic importance. Rapid and species-specific detection is essential for effective clinical management and epidemiological surveillance. This study aimed to develop and diagnostically evaluate two novel species-specific loop-mediated isothermal amplification (LAMP) assays for the direct detection of D. immitis and D. repens in canine whole blood, performed in parallel in separate reactions, with emphasis on simplified and potentially near-point-of-care applicability. Primers targeting mitochondrial COI and NADH gene regions were designed and validated. In silico specificity analysis against 13 filarioid species confirmed the absence of non-specific primer binding. A direct closed-tube LAMP protocol using sodium hydroxide–Chelex-100 lysis was optimized, enabling amplification without conventional DNA extraction while reducing contamination risk and processing time to under 60 min. Relative diagnostic performance was evaluated relative to quantitative real-time PCR (qPCR) results. Using purified DNA, the D. repens assay achieved 100% relative sensitivity and relative specificity, whereas the D. immitis assay showed 94.5% relative sensitivity and 100% specificity. In direct whole-blood assays, relative specificity remained 100% for both targets, while sensitivity decreased to 90.9% for D. immitis and 77.42% for D. repens, with most false-negative reactions associated with high qPCR Ct values (>30). These findings demonstrate that the proposed assays provide a rapid and practical molecular diagnostic approach with potential applicability for point-of-care veterinary testing. Full article

Forensic biotechnology is a rapidly evolving interdisciplinary field integrating molecular biology, genomics, and data science to address complex investigative challenges. Its applications span diverse domains, including criminalistics, food authentication, environmental monitoring, and bioterrorism preparedness. Advanced technologies such as Next-Generation Sequencing (NGS), CRISPR-Cas biosensors, and Artificial Intelligence (AI) play pivotal roles in modern diagnostics. NGS and eDNA revolutionize genetic profiling and ecological tracking, while microbiome analysis provides crucial insights into post-mortem intervals, cause of death, and geolocation. Simultaneously, CRISPR-based methods enable ultra-rapid pathogen detection, nanobiotechnology facilitates portable Lab-on-a-Chip (LOC) DNA analysis, and AI-driven algorithms optimize the interpretation of complex genomic mixtures and epigenetic age estimation. Despite these breakthroughs, significant challenges persist, including the strict legal admissibility of novel methodologies, the “black-box” dilemma in AI, ethical concerns regarding genetic privacy, and the critical need for global standardization. This review critically examines current biotechnological progress and future prospects, emphasizing the necessity of interdisciplinary collaboration to ensure reliable, accurate, and ethically sound forensic practices. Full article

Background: The rapid rise in antimicrobial resistance (AMR) represents one of the most pressing global health challenges of the 21st century, threatening antibiotic effectiveness, compromising clinical outcomes, and undermining healthcare systems. Understanding how resistant pathogens emerge and spread across human, animal, and environmental sectors is essential for effective global response. Main body: This review evaluates traditional and advanced AMR detection methodologies, including phenotypic assays, molecular diagnostics, whole-genome sequencing (WGS), metagenomics, and biosensor-based technologies. It also highlights the role of bioinformatics tools, surveillance databases, and integrated platforms that support real-time analysis. Genomic surveillance provides unparalleled resolution for characterizing resistance mechanisms, transmission patterns, and evolutionary trajectories of multidrug-resistant organisms. Techniques such as WGS and metagenomics allow timely and precise identification of resistance genes, improving outbreak detection and strengthening antimicrobial stewardship. Despite these advantages, the adoption of genomic surveillance faces barriers in low- and middle-income countries, including high costs, limited infrastructure, insufficient technical expertise, and the lack of standardized data frameworks. Conclusions: Genomic surveillance is a transformative tool for combating AMR and strengthening global health systems. Effective implementation requires sustained investment, capacity-building, coordinated cross-sector collaboration, and commitment to the One Health approach to ensure equitable access and long-term global impact. Full article

Background: The rapid expansion of functional ready-to-drink (RTD) beverages—formulated with prebiotic fibers, botanical extracts, and reduced sugar—has outpaced systematic characterization of their small-molecule composition. Methods: We applied dual-mode untargeted high-resolution liquid chromatography–tandem mass spectrometry (LC–MS/MS), integrating hydrophilic interaction (HILIC) and reversed-phase C18 separations, to profile five commercial RTD beverages spanning distinct formulation categories: Coca-Cola^®, Poppi^® Orange, OLIPOP^® Cream Soda, Pure Leaf^® Unsweetened Black Tea, and BeePop™ Peach + Orange Blossom Honey. Results: Across all products, 478 compounds were structurally annotated at Metabolomics Standards Initiative (MSI) Levels 1 and 2, of which 42 matched compounds with reported bioactivity in a curated literature-based reference database. Seventeen compounds—including the NAD+ precursor trigonelline and multiple B vitamins—were detected across all five products. The number and diversity of compounds with reported bioactivity varied substantially by product and correlated with botanical ingredient complexity. Conclusions: This work presents a qualitative molecular survey of the RTD beverage category using standardized, dual-mode untargeted metabolomics, providing a reference dataset for future targeted quantitation studies. Full article

Green mold disease caused by Trichoderma species represents one of the most serious threats to the cultivation of Ganoderma lingzhi. In this study, a rapid molecular detection system was developed for the identification of green mold caused by Trichoderma longibrachiatum. The pathogen was first identified based on morphological characteristics and pathogenicity tests. Species-specific primers were then designed targeting the RPB2 gene region, and their specificity and sensitivity were evaluated using polymerase chain reaction (PCR). The results confirmed that the causal pathogen was T. longibrachiatum. The designed primers exhibited high specificity and produced amplification only with genomic DNA from the target strain. Sensitivity assays demonstrated that the minimum detectable DNA concentration was 10⁻² ng/µL. In soil inoculation experiments, the target DNA could be detected in soil containing 4.28 × 10⁶ spores/g using the developed primers. Compared with conventional culture-based detection methods, the molecular detection system established in this study is faster, more accurate, and highly sensitive. This method provides a practical diagnostic tool for the early detection and management of green mold disease in G. lingzhi cultivation. Full article

Streptococcus agalactiae is a major contagious pathogen of bovine mastitis and causes substantial economic losses in the dairy industry. In this study, a dual-readout RPA-PfAgo detection platform targeting the conserved cfb gene of S. agalactiae was established and optimized. Seven pairs of RPA primers were designed and screened to construct the Basic-RPA assay, and six guide DNAs (gDNAs) together with a specific probe were evaluated for PfAgo-assisted detection. Field validation was performed using 153 bovine milk samples collected from five dairy-farming regions in China, and assay performance was compared with bacteriological culture and a standardized quantitative PCR (qPCR) assay. The Basic-RPA assay achieved optimal amplification at 37 °C for 30 min, with a detection limit of 1 × 10⁻³ ng/µL and no cross-reactivity with non-target bacteria. The optimized RPA-PfAgo-RTF assay detected as few as 10 copies/µL, whereas the RPA-PfAgo-LFD assay detected 100 copies/µL, and both formats showed high analytical specificity. In field milk samples, bacteriological culture detected 21 positive samples, whereas standardized real-time PCR (qPCR), RPA-PfAgo-RTF, and RPA-PfAgo-LFD each detected 33 positive samples. When compared with bacteriological culture as a conventional comparator, all three molecular assays showed 100.00% positive agreement, 90.91% negative agreement, and a Kappa value of 0.733. In addition, RPA-PfAgo-RTF and RPA-PfAgo-LFD were completely concordant with the standardized qPCR assay across all 153 samples. These results indicate that the dual-readout RPA-PfAgo platform is a rapid and reliable molecular tool for detection of S. agalactiae in bovine milk. Full article