Search Results (796)

‘Genetic zipper’ technology—based on CUAD (Contact Unmodified Antisense DNA) biotechnology, briefly CUADb—represents a step forward in eco-friendly pest control. This unique innovative approach is based on a fundamentally new biological mechanism—a two-step DNA containment (DNAc) mechanism. DNAc employs short, unmodified antisense DNA molecules to selectively degrade target pre-rRNA and/or rRNA in insect pests recruiting up-regulated RNase H1 and RT-RNase H during DNAc, disrupting protein synthesis and causing the down-regulation of kinases due to ATP insufficiency and ultimately leading to high mortality rates. Demonstrating exceptional speed and precision, this technology enables the design of effective and selective DNA pesticides (oligonucleotide pesticides) for no less than 15% of known insect pests in a single day. In this review, we highlight the simplicity and global applicability of this technology using case studies involving 12 economically significant pest species, including hemipterans and one spider mite, from five continents. These oligonucleotide pesticides, computationally predicted via the DNAInsector web tool, are supposed to offer 80–90% efficacy against target pests within one–two weeks under laboratory or field conditions. Their action is primarily non-systemic, requiring direct contact. Oligonucleotide pesticides are environmentally safe, biodegradable, and highly specific, reducing risks to non-target organisms. The ‘genetic zipper’ technology not only provides a powerful tool for researchers and practitioners but also opens a new era in DNA-programmable pest management, where personalized, algorithm-driven pesticides can be easily created and applied for sustainable agriculture. Full article

Neisseria gonorrhoeae is a major sexually transmitted pathogen and global public health concern. A porA pseudogene-targeting TaqMan quantitative PCR (qPCR) assay was evaluated as a complementary laboratory-based method for the qualitative detection of N. gonorrhoeae and exploratory Z017-calibrated molecular estimation in cultured reference strains. Two candidate probe sets were evaluated, and the probe set generating a 157 bp amplicon was selected for a defined two-step qPCR workflow with a 52 min on-instrument amplification runtime. In silico analysis showed compatible target site arrangements in all 201 N. gonorrhoeae genomes and none in 114 non-target organisms. The assay detected N. gonorrhoeae DNA down to 1 × 10⁻⁵ ng/µL, with no amplification of the tested non-target organisms. All 23 N. gonorrhoeae reference strains and 80 N. gonorrhoeae-positive clinical specimens were detected, whereas no amplification was observed in the 20 negative clinical specimens, using DNA extracted from the primary clinical specimens. A standard curve was generated from the titered Z017 reference material (R² = 0.999; efficiency, 94.6%), which enabled reproducible qPCR-derived CFU/mL-equivalent molecular estimation in cultured World Health Organization reference strains. The qPCR-derived estimates were lower than the paired culture-derived CFU/mL values, and the paired log10-transformed values did not show a statistically significant correlation. These findings provide an assay design and validation-oriented laboratory-based framework combining in silico sequence assessment, the qualitative detection of N. gonorrhoeae in clinical specimens, and exploratory molecular estimation in cultured reference strains, supporting future assay development and bacterial burden research. Full article

Yellow adhesive chromotropic traps are widely deployed across Mediterranean olive cultivation to monitor B. oleae population dynamics, yet their adhesive surfaces can also retain non-target organisms. Here we report the first documented entrapment of the common pipistrelle Pipistrellus pipistrellus (Schreber, 1774) on an adhesive monitoring trap in Greece, recorded on 15 May 2026 in a traditionally managed olive grove on Lesvos Island (North Aegean). The adult specimen (wingspan of 23.1 cm, body mass of 5.9 g) was independently identified by two specialist chiropterologists and confirmed dead by direct examination. The trap surface bore small-bodied Diptera belonging to groups known to occur in the diet of P. pipistrellus, raising the possibility that prey-associated cues may have contributed to the encounter. We argue that this record exposes an agroecological contradiction relevant to conservation-oriented olive management: the monitoring device deployed to track B. oleae may incidentally eliminate an insectivorous bat belonging to the broader natural enemy community associated with olive agroecosystems. Given the extent of adhesive trap deployment across Mediterranean olive cultivation and the strict legal protection afforded to all European bats under the Habitats Directive (92/43/EEC), systematic bycatch recording, mechanistic investigation and evaluation of simple mitigation measures are warranted. Full article

Ips hauseri Reitter is the most important wood-boring pest of natural Tianshan spruce (Picea schrenkiana) forests and is listed as an A2 quarantine pest by the European and Mediterranean Plant Protection Organization (EPPO). Because the beetle is small and cryptic and its immature stages cannot be reliably identified by morphology, a rapid molecular identification method is urgently needed. Here, we targeted the mitochondrial cytochrome c oxidase subunit I (COI) gene, designed a set of I. hauseri-specific LAMP primers, and established a visually interpretable LAMP assay using a WarmStart colorimetric system. The optimal reaction conditions were 63.8 °C for 35 min. The assay amplified only I. hauseri, with no cross-reaction with the related species Pityogenes spessivtsevi, Ips typographus, Ips subelongatus, and Hylurgus ligniperda; its limit of detection was 100 pg of genomic DNA; and it amplified efficiently from adults, larvae, pupae, and a single egg. Positive results could be scored directly by eye from the red-to-yellow colour change in the reaction, requiring no precision instruments at any step. Using a non-destructive alkaline crude-DNA extraction (HotSHOT), the assay also amplified I. hauseri directly from single adults and larvae, confirming its compatibility with a rapid, field-simple sample preparation. The LAMP assay was rapid, specific within the tested panel, and simple. However, broader validation with additional sympatric non-target species and geographically distinct populations of I. hauseri is needed before routine field or quarantine application. Full article

Tropical fruit processing generates substantial quantities of seed-derived by-products that remain underutilized despite their potential as sources of bioactive compounds, proteins, lipids, dietary fiber, and polysaccharides. This critical narrative review evaluates their conversion into functional ingredients by integrating compositional variability, pretreatment and extraction, antinutritional-factor mitigation, structural characterization, food-system performance, storage stability, gastrointestinal behavior, safety, regulatory readiness, and scale-up feasibility. Direct evidence was distinguished from mechanistic support derived from non-target botanical matrices. An exploratory semantic clustering of 100 PubMed records published in 2024–2026 was conducted with Carrot2 to organize application-oriented themes and identify knowledge gaps; its output was interpreted as a thematic aid rather than a quantitative trend analysis. The review also examines biochar production from exhausted or non-food-grade seed biomass as a complementary cascading route, considering feedstock properties, thermochemical conditions, physicochemical quality, safety, application performance, and techno-economic constraints. The main contribution is an evidence-derived multi-stage decision framework linking ingredient validation with residual-biomass valorization. Current evidence supports promising nutritional and technological applications but remains heterogeneous and predominantly laboratory-based, with limited standardized comparisons, in vivo validation, regulatory assessment, and pilot-scale studies. Advancing these materials therefore requires application-specific, safety-oriented, and economically realistic validation within circular bioeconomy systems. Full article

Pythium insidiosum, the causative agent of pythiosis, remains an important pathogen in tropical and subtropical regions, where delayed diagnosis is associated with severe disease and poor clinical outcomes. We developed a loop-mediated isothermal amplification coupled with lateral flow dipstick (LAMP-LFD) assay for the rapid detection of P. insidiosum. The assay employs dual-labeled LAMP amplicons that are directly detected by a lateral flow dipstick, enabling simple visual interpretation without agarose gel electrophoresis. Analytical performance was evaluated using genomic DNA from 102 clinically relevant isolates, comprising 51 P. insidiosum isolates and 51 non-target organisms. The LAMP-LFD assay correctly detected all P. insidiosum isolates, achieving an inclusivity of 100.0% (95% CI: 93.0–100.0%), and correctly excluded 48 of 51 non-target organisms, yielding an exclusivity of 94.1% (95% CI: 83.8–98.8%). The assay demonstrated a detection limit of 10 fg of genomic DNA per reaction, representing at least 1000-fold greater analytical sensitivity than the reference multiplex PCR (m-PCR) assay. Clinical validation using 47 blinded animal tissue specimens showed a sensitivity of 90.3%, specificity of 87.5%, accuracy of 89.4%, and good agreement with the reference standard (culture and ITS sequence analysis) (κ = 0.76). In comparison, the m-PCR assay achieved a sensitivity of only 41.9%, indicating substantially lower clinical detection performance. In conclusion, direct LFD-based detection preserves the analytical performance of the established P. insidiosum LAMP assay while providing a clear and easily interpretable visual endpoint. The LAMP-LFD assay represents a rapid, sensitive, and practical tool for screening pythiosis, offering improved diagnostic performance and more reliable result interpretation. Full article

The use of environmental nucleic acid (eNA), both DNA and RNA, as a means for surveillance has been a fixture in the scientific literature for many years. The application of environmental screening for genomic signatures of organisms of interest—particularly those of diagnostic concern—is a promising yet still under-utilised tool for sample screening. While the literature tends to focus on the use of high-throughput sequencing (HTS) to detect organisms of interest using metagenomic or metatranscriptomic sampling, this approach is not cost-competitive with more traditional targeted molecular test methods. Consequently, eNA sampling still has not gained significant traction in practical settings despite its popularity in ecological research. To address these issues in a biosecurity context, we report here the development of a testing protocol to monitor irrigation water for the presence of pepino mosaic virus (PepMV) that also includes an endogenous Sphingomonas control. We employed passive sampling through the immersion of filtering devices into the water system at three commercial growing operations at two time points to collect samples with minimal hands-on effort, while simultaneously developing and validating molecular methods for the recovery of RNA competent for both PCR and high-throughput sequencing. We demonstrate not only the ability to detect PepMV from water collections, but also that the method is robust to the accumulation of non-target material and does not lose signal if viruses are only transiently present in the water system. Finally, we developed a capsid-integrity pre-treatment protocol for differentiating between intact and denatured (non-viable) virus particles during PCR testing. This work presents a low-cost and low-effort technique for proactive screening of commercial greenhouse facilities to facilitate early detection of harmful crop pests and pathogens. Full article

Phenolic acids and flavonoids are major bioactive constituents of Sparganium stoloniferum tubers (SL), yet their spatial organization within this medicinal organ remains insufficiently characterized, limiting understanding of how anatomical structure relates to metabolite distribution. To address this, we integrated mass spectrometry imaging (MSI), non-targeted LC–MS and targeted LC–MS/MS metabolomics, and reverse transcription quantitative polymerase chain reaction (RT–qPCR) analysis to characterize the spatial patterns of phenylpropanoid- and auxin-related metabolites and selected transcripts in SL. MSI and non-targeted LC–MS profiling showed that phenylpropanoid- and flavonoid-related metabolites were preferentially accumulated in the peripheral cortex rather than in the stele, although the stele constitutes the dominant internal tissue of the tuber. Targeted LC–MS/MS further confirmed that representative hydroxycinnamic acids and caffeoylquinic acid derivatives were enriched in the cortex and, for most validated metabolites, in the cell wall-enriched fraction. Notably, spatial metabolomic profiling also revealed a contrasting stele-biased distribution of indole-related metabolites, including indole-3-acetamide-related features detected by MSI and non-targeted LC–MS and indole-3-acetic acid (IAA) enrichment validated by targeted LC–MS/MS. RT–qPCR analysis showed that phenylpropanoid biosynthetic genes were generally more highly expressed in the cortex and cell wall-enriched fraction, whereas auxin-related genes showed higher expression in the stele. Together, these results show cortex-biased phenylpropanoid accumulation, preferential association of most validated phenylpropanoid-related metabolites with the cell wall-enriched fraction, and stele-associated IAA accumulation and auxin-related transcript expression. This study provides a spatial framework for understanding metabolite partitioning in medicinal aquatic storage organs and highlights the importance of integrating anatomical, metabolomic, and gene expression information in medicinal plant research. Full article

Background: The fall armyworm, S. frugiperda, is a globally threatening insect pest of agricultural crops, including in Thailand. Symbiotic bacteria in the genera Xenorhabdus and Photorhabdus, which are derived from entomopathogenic nematodes, can produce diverse bioactive compounds with insecticidal activities. However, no study on Thai isolates of these symbiotic bacteria against S. frugiperda has been reported, leading to a significant gap in local biopesticide development. Methods: Two symbiotic bacterial isolates, X. miraniensis (bMH16.1_TH) and P. akhurstii (bSBR11.1_TH), were isolated from Thai entomopathogenic nematodes and cultured in LB broth at 10⁸ CFU/mL. Three bacterial fractions, i.e., whole-cell suspension, cell supernatant, and cell pellet, were prepared from each isolate. Insecticidal bioassays were conducted by topical application on second- and fifth-instar larvae of S. frugiperda (10 larvae × 5 replicates per treatment, CRD). Larval mortality was recorded daily for seven days, and median lethal time (LT₅₀) was calculated. Data were analyzed by two-way ANOVA with Duncan’s Multiple Range Test (p < 0.05). Results: The results indicated that both bacterial isolates caused significantly higher larval mortality than controls across all treatments (p < 0.05). X. miraniensis (bMH16.1_TH) showed higher virulence, achieving ~80% mortality of second-instar larvae within 24 h and 100% by day three. Whole-cell suspensions consistently produced the fastest and highest mortality with the lowest LT₅₀ values in both larval stages. Second-instar larvae were significantly more susceptible than fifth-instar larvae. The shortest LT₅₀ was recorded in second-instar larvae treated with X. miraniensis supernatant at 0.616 days, while fifth-instar larvae generally showed higher LT₅₀ values across all treatments. Conclusions: Our comparative study demonstrated that the insecticidal efficacy varied between the two symbiotic bacterial isolates and among their fractions. Whole-cell suspensions of X. miraniensis (bMH16.1_TH) produced the highest larval mortality and the smallest LT₅₀ values, particularly against early-instar larvae. These findings demonstrated that local symbiotic bacteria are promising candidates to be developed as sources of biopesticides against this insect pest. In addition, characterizing active insecticidal compounds, evaluating field efficacy, and assessing safety to non-target organisms need to be further studied to support integration of these biopesticides in sustainable pest management programs. Full article

Glyphosate is one of the most widely used herbicides worldwide, and its extensive application has raised increasing concern regarding environmental occurrence, ecological exposure, and potential risks to non-target organisms. Its major degradation product, aminomethylphosphonic acid (AMPA), has also received growing attention because of its persistence, transport behavior, and contribution to long-term contamination profiles. To clarify the development and emerging priorities of this field, we conducted a bibliometric review of glyphosate-related publications indexed in the Web of Science Core Collection from 1974 to 2024. After screening and data cleaning, 7050 articles and reviews were included. Publication output increased markedly over time, with annual publications exceeding 300 after 2019 and reaching a peak of 488 in 2023. The United States ranked first with 1928 publications, accounting for 27.4% of the total output, followed by Brazil and China. Keyword co-occurrence, temporal overlay, and collaboration analyses showed that glyphosate research has shifted from early agronomic topics, including herbicide efficacy, crop selectivity, and resistance management, toward a broader environmental research framework. Three major research fronts were identified: mechanism-oriented ecotoxicology in non-target organisms, environmental fate and transport of glyphosate and AMPA across soil–water–sediment systems, and the development of analytical and sensing technologies for environmental monitoring. The results further indicate that the field is moving from single-compound residue assessment toward integrated contaminant-ecology perspectives linking occurrence, transformation, biological response, exposure assessment, and ecological risk. Future studies should strengthen the integration of long-term environmental monitoring, AMPA-inclusive risk assessment, realistic multi-stressor exposure scenarios, and field-deployable detection technologies. This review provides a quantitative overview of the global research landscape and identifies priority directions for environmental assessment and management of glyphosate and AMPA contamination. Full article

Fenthion is a veterinary pharmaceutical (VP) used in the control of ecto- and endoparasites in livestock. This active substance may reach the soil through the animals’ urine and feces, but the ecotoxicological data of this compound on non-target soil fauna is limited. This study aimed to evaluate the chronic ecotoxicity of a fenthion-based veterinary formulation on six species of soil invertebrates. Ecotoxicological tests were performed in a natural tropical soil (Entisol) with earthworms Eisenia andrei, enchytraeids Enchytraeus crypticus and Enchytraeus bigeminus and collembolans Folsomia candida, Sinella curviseta and Proisotoma minuta, following ISO protocols. The species sensitivity distribution (SSD) approach was employed to establish protective concentrations (PCs) for fenthion in soil. Collembolans were the most sensitive organisms; the EC50 for F. candida, S. curviseta and P. minuta were 0.72, 0.93 and 0.48 mg kg⁻¹, respectively. Earthworms E. andrei presented intermediate sensitivity (EC50 = 28.40 mg kg⁻¹). Enchytraeids were the least sensitive animals (EC50 for E. crypticus and E. bigeminus of 233.66 and 221.47 mg kg⁻¹, respectively). SSD-derived PCs for 95%, 90%, 80% and 50% of soil species were estimated in 0.017, 0.054, 0.225 and 3.432 mg kg⁻¹, respectively. The predicted environmental concentration (PEC) of fenthion in soil via veterinary use (0.032 mg kg⁻¹) is almost twice the PC95 value (0.017 mg kg⁻¹), indicating potential ecological risk of fenthion to soil species. This study brings new information regarding the ecotoxicological profile and ecological risk of a relevant VP to the edaphic community. Full article

The fall armyworm (FAW), Spodoptera frugiperda, is a highly polyphagous pest that has rapidly expanded across Africa, Asia, and Oceania, threatening food security and agricultural productivity. Reliance on synthetic insecticides for FAW management is increasingly challenged by insecticide resistance, environmental contamination, and adverse effects on non-target organisms and human health. This review synthesizes current knowledge on the potential of endophytic fungi as a sustainable strategy for FAW management. Following a systematic literature search, 59 original research articles published over the past 41 years were evaluated. These studies investigated 44 fungal species across 37 host crops, with Zea mays representing the most extensively studied system. The fungal species most frequently assessed were Beauveria bassiana, Metarhizium anisopliae sensu lato (s.l.), and Epichloe coenophialum. Overall, several fungal species successfully colonized plant tissues and reduced FAW performance through increased larval mortality, delayed development, and reduced feeding. These effects were associated with the production of bioactive secondary metabolites, induction of plant defense responses, and alteration of volatile organic compound emissions. Despite these promising findings, important knowledge gaps remain. Studies assessing effects on non-target organisms and natural enemies are scarce, and field validation remains limited. Research efforts are also geographically biased, with relatively few studies conducted in Africa despite the continent experiencing some of the highest FAW-related losses. Future research should prioritize field evaluations, multitrophic interaction studies, investigations in underrepresented regions, and socio-economic analyses to support farmer adoption. Addressing these gaps will be critical for determining the practical role of endophytic fungi in sustainable FAW management. Full article

Pit mud (PM) harbors abundant bacteria and archaea, shaping Baijiu’s flavor and adapting to high-ethanol and acidic conditions. This study integrated 16S rRNA sequencing, physicochemical analysis, and non-targeted metabolomics. The results showed that the 30-year-old Zhuyechun PM had higher levels of humic acid, while the 20-year-old Fengchun and Bitanchun PM exhibited near-neutral pH, which is favorable for Clostridia and methanogens. Canonical correspondence analysis revealed Rummeliibacillus and Caproiciproducen positively correlated with organic carbon, humic acid, and available phosphorus, whereas Methanosarcina showed negative correlations. In Baijiu, 1206 non-volatile signals were identified, with unsaturated fatty acid biosynthesis as a key flavor pathway. Methanogens significantly associated with key metabolites (p < 0.05). These findings reveal the correlation between the PM microecosystem and Baijiu quality, providing insights for the development of high-quality Baijiu. Full article

Plant cell, tissue, and organ cultures are increasingly used as controlled platforms for producing pharmaceutically important specialized metabolites. Their productivity can be improved through elicitors, plant biostimulant-like substances, microbial inputs, and nano-enabled delivery systems. However, the regulatory interpretation of these inputs is not determined only by their biological mechanism. It also depends on product composition, formulation, intended use, exposure scenario, and label claim. This review analyzes how elicitors, plant biostimulants, biofertilizer- and bioinput-related concepts, and nano-enabled inputs are positioned within selected regulatory frameworks in the European Union, United States, Canada, Brazil, India, China, and Mexico. The analysis shows that current frameworks increasingly emphasize product identity, efficacy substantiation, safety evidence, and labeling, but they remain fragmented for multifunctional materials that may simultaneously affect nutrient-related processes, abiotic stress tolerance, defense signaling, growth regulation, and specialized metabolism. Nano-enabled and encapsulated formulations create additional challenges because they can modify uptake, persistence, bioavailability, release behavior, residue profiles, and non-target exposure compared with conventional formulations. A clearer distinction is therefore needed between contained biotechnological use in plant biofactories and open-environment agricultural applications. Future regulatory development should move toward formulation-level assessment, claim-specific evidence, nanospecific characterization, traceability, and harmonized terminology to support innovation while ensuring safety, reproducibility, and regulatory clarity. Full article

Glyphosate (GLY), one of the most extensively applied broad-spectrum herbicides worldwide, frequently contaminates soil and aquatic ecosystems, posing serious threats to human health, non-target organisms, soil microbial communities, and environmental sustainability. In the present study, phytoremediation using Medicago sativa (MS) was evaluated for the removal of GLY from contaminated soil under greenhouse conditions, with remediation efficiency enhanced through inoculation with two bacterial bioagents, Bacillus sp. h10 (BS) and Pseudomonas aeruginosa KZFS4 (PA). Biochemical parameters, including superoxide dismutase (SOD), catalase (CAT), hydrogen peroxide (H₂O₂), and malondialdehyde (MDA), together with detoxification-related gene expression, were investigated in the roots and leaves of MS exposed to GLY stress. The combined application of MS with BS + PA, followed by MS + PA and MS + BS, significantly decreased GLY residues in soil and increased GLY accumulation in plant roots and leaves after 1, 3, 7, and 10 days compared with MS treatment alone. In vitro batch equilibrium experiments demonstrated that BS and PA desorbed 33.63 and 40.56 µg g⁻¹ of GLY, respectively, thereby enhancing its removal from soil. The persistence of GLY was highest in contaminated soil without treatment, exhibiting a half-life (t₁/₂) of 52.66 days, whereas the shortest half-life (6.69 days) was recorded in soil treated with MS combined with BS and PA relative to sterilized contaminated soil. Furthermore, inoculation with BS and PA markedly increased SOD and CAT activities in MS tissues, while significantly reducing H₂O₂ and MDA accumulation, indicating alleviation of oxidative stress. GLY exposure also triggered substantial upregulation of detoxification-associated genes, including cytochrome P450, glutathione S-transferases (GST), glycosyltransferases (GTs), and ABC transporters in MS. These findings demonstrate that the integration of BS and PA with phytoremediation effectively accelerates GLY dissipation and reduces pesticide-associated toxicity in contaminated soils and plants. Full article