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59 pages, 2944 KB  
Review
Boron’s Double Edge—Antibiotics, Toxins, and the Fine Line Between Them
by Valery M. Dembitsky, Alexander O. Terent'ev, Sergey V. Baranin and Ion Romulus Scorei
Molecules 2026, 31(6), 1021; https://doi.org/10.3390/molecules31061021 - 18 Mar 2026
Viewed by 178
Abstract
Boron is a chemically distinctive bioelement whose electron-deficient structure enables reversible coordination with oxygen-rich functional groups such as diols and hydroxyls. This property allows boron to modulate molecular stability, conformation, and biological reactivity, giving rise to both beneficial pharmacological effects and toxicological outcomes. [...] Read more.
Boron is a chemically distinctive bioelement whose electron-deficient structure enables reversible coordination with oxygen-rich functional groups such as diols and hydroxyls. This property allows boron to modulate molecular stability, conformation, and biological reactivity, giving rise to both beneficial pharmacological effects and toxicological outcomes. This review examines the dual biological role of boron through the framework of bioactive boron-containing natural products and natural compounds capable of forming reversible boron complexes. Particular attention is given to naturally occurring boron-containing antibiotics, including the polyketide macrodiolides boromycin, aplasmomycin, tartrolons, and hyaboron, where boron plays a direct structural and functional role in antimicrobial activity. These compounds demonstrate how boron coordination can influence ion transport, membrane interactions, and molecular assembly, contributing to potent antibacterial properties. Beyond intrinsically boron-containing metabolites, many natural antibiotics and toxins possess oxygen-rich architectures capable of forming transient borate complexes through vicinal 1,2-diol motifs. Examples include polyene macrolide antibiotics such as amphotericin B, fungichromin, and nystatin, as well as tetracyclines, rifamycins, and macrolides such as sorangicin A, where boron coordination may affect solubility, aggregation, ionophoric behavior, and biological selectivity. Similar chemistry is observed in marine neurotoxins and polyether toxins—including tetrodotoxin, saxitoxin derivatives, azaspiracids, pectenotoxins, ciguatoxins, and gambierones—whose hydroxyl-rich frameworks enable reversible interactions with boron species present in seawater. Such complexation may enhance aqueous stability and contribute to trophic transfer and bioaccumulation within marine ecosystems. By framing boron as a molecular “double edge,” this review integrates chemical, biological, and environmental perspectives to highlight how boron coordination can simultaneously enhance antimicrobial activity while influencing toxicity and ecological persistence. Recognizing the role of boron in shaping the activity of natural products provides new insight into antibiotic function, toxin behavior, and the broader impact of boron chemistry in biological systems. Full article
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18 pages, 2570 KB  
Article
Functional Divergence and Toxin Coupling of Cyanobacterial Blooms Across the Lake–River Continuum: Insights from the Lake Taihu Watershed
by Xiang Wan, Yucong Li, Qingju Xue, Guoxiang Wang and Liqiang Xie
Toxins 2026, 18(2), 89; https://doi.org/10.3390/toxins18020089 - 9 Feb 2026
Viewed by 472
Abstract
While harmful cyanobacterial blooms (HCBs) are extensively characterized in eutrophic lakes, the ecological dynamics of connected river networks remain oversimplified, obscuring the mechanisms of community assembly and toxin distribution across the lake–river interface. This study investigated the spatial heterogeneity of HCBs and microcystins [...] Read more.
While harmful cyanobacterial blooms (HCBs) are extensively characterized in eutrophic lakes, the ecological dynamics of connected river networks remain oversimplified, obscuring the mechanisms of community assembly and toxin distribution across the lake–river interface. This study investigated the spatial heterogeneity of HCBs and microcystins (MCs) in the Lake Taihu watershed, revealing a complex functional divergence between lotic and lentic ecosystems. The rivers functioned as primary nutrient sources, with Total Nitrogen (3.35 ± 1.52 mg·L−1) and Total Phosphorus (0.21 ± 0.22 mg·L−1) concentrations being 1.7-fold and 1.8-fold higher, respectively, than those in the lake during peak periods. Conversely, the lake acted as a biological sink, supporting a peak cyanobacterial density (3.32 × 107 cells·L−1) nearly 1.5 times that of the river network. Phytoplankton community analysis revealed distinct ecological niches: while the lentic lake environment was almost exclusively dominated by colonial Microcystis (>90% relative abundance), the lotic river networks harbored a diverse assemblage with significant contributions from filamentous Oscillatoria and Dolichospermum. Correspondingly, intracellular MC (IMC) in the lake (up to 14.5 μg·L−1) significantly exceeded riverine levels (generally <1.0 μg·L−1). Despite these compositional differences, toxin dynamics exhibited strong bidirectional coupling (r > 0.75, p < 0.01), suggesting a spillover effect where the lake determines the watershed’s toxin burden during rivers outflow period. Redundancy Analysis (RDA) further elucidated that limnetic blooms were primarily regulated by water temperature and pH, whereas riverine communities were strictly constrained by hydrodynamic flow. Consequently, the health risk assessment revealed a highly heterogeneous landscape where, beyond the northern lake bays, specific semi-lentic river segments emerged as cryptic hotspots. These findings demonstrate that while nutrients fuel the system, hydrodynamic conditions act as the ultimate ecological filter determining the spatiotemporal distribution of cyanobacterial risks, necessitating an integrated approach to monitoring the entire lake–river continuum. Full article
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23 pages, 962 KB  
Article
Virulence Gene Profiles of Extended-Spectrum β-Lactamase (ESBL)-Producing Escherichia coli Isolated from Turkeys in Hungary: A Whole-Genome Sequencing Study
by Ádám Kerek, Ábel Szabó, Gergely Tornyos, Eszter Kaszab, Krisztina Bali and Ákos Jerzsele
Vet. Sci. 2025, 12(12), 1141; https://doi.org/10.3390/vetsci12121141 - 29 Nov 2025
Viewed by 711
Abstract
Background: Escherichia coli strains associated with poultry are increasingly recognized as reservoirs of both virulence and resistance genes, posing significant zoonotic risks throughout the food production chain. However, the genotypic architecture and pathogenic potential of isolates from large-scale turkey farms remain under [...] Read more.
Background: Escherichia coli strains associated with poultry are increasingly recognized as reservoirs of both virulence and resistance genes, posing significant zoonotic risks throughout the food production chain. However, the genotypic architecture and pathogenic potential of isolates from large-scale turkey farms remain under characterized, particularly in the context of extended-spectrum β-lactamase (ESBL) production. Methods: A total of 160 ESBL-producing E. coli isolates were collected from healthy turkeys on intensive Hungarian farms. Whole genome sequencing (WGS) was performed to characterize virulence factors. Functional annotation included screening for fimbrial adhesins, iron acquisition systems, secretion pathways, and autotransporter toxins, using VirulenceFinder and Prodigal-based genome annotations. Data analysis included assembly quality control with QUAST and BUSCO, and comprehensive virulome profiling. Results: The isolates exhibited a functionally diverse virulence profile encompassing classical ExPEC-associated colonization factors (type I, P, S fimbriae; curli; ECP), multiple iron acquisition systems (enterobactin, salmochelin, aerobactin, yersiniabactin, and heme uptake), and key secretion systems (LEE-associated T3SS and T2SS). Genetic hallmarks of avian pathogenic E. coli (APEC), uropathogenic pathogenic E. coli (UPEC), and enteropathogenic E. coli (EPEC) pathotypes co-occurred in 44% of the isolates, indicating a mosaic virulence landscape. Notably, serine protease autotransporters of Enterobacteriaceae (SPATE) toxins (Vat, Pic) and ColV-type plasmid-associated modules were frequently detected. All isolates were confirmed by ESBL producers, highlighting their antimicrobial resistance potential. Conclusions: This study reveals that E. coli strains isolated from turkeys possess a complex, host-adapted virulence repertoire capable of supporting both enteric and extraintestinal infections. The co-occurence of APEC-, UPEC-, and EPEC-like traits—combined with ESBL production—underscores their One Health relevance. These findings support the need for host-specific surveillance, functional validation, and integrative control strategies in poultry systems. Full article
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20 pages, 745 KB  
Review
Transboundary Diseases and One Health Approach Implications for Global Health Threats, with Particular Interest in Conservation and Bioterrorism
by Massimo Giangaspero, Salah Al Mahdhouri, Sultan Al Bulushi, Metaab K. Al-Ghafri and Pasquale Turno
Pathogens 2025, 14(12), 1193; https://doi.org/10.3390/pathogens14121193 - 22 Nov 2025
Viewed by 1396
Abstract
Among animal diseases, those characterized with transboundary potential enhance their interconnection to the One Health principle. Zoonoses with a higher capacity to spread compared to other diseases with a lower level of transmissibility multiply their potential impact on human populations. The routes and [...] Read more.
Among animal diseases, those characterized with transboundary potential enhance their interconnection to the One Health principle. Zoonoses with a higher capacity to spread compared to other diseases with a lower level of transmissibility multiply their potential impact on human populations. The routes and speed of transmission and virulence may also increase the impact on animal health in the zootechnic sector and in wild animals. This risk, especially in endangered species, has the potential to alter biodiversity, negatively affecting the environment. The characteristics of these pathogens represent a global health danger that requires knowledge and the capacity for prevention and control, considering the possibility of natural outbreak occurrence together with the deliberate use of such pathogens as biological weapons for terrorist attacks. Animal pathogens, particularly those with zoonotic potential, have long been considered for use in bioterrorism. International conventions prohibit the use of microbiological and toxin weapons. Furthermore, recent European legislation has also addressed the potential misuse of animal pathogens in bioterrorism. In this context, the Parliamentary Assembly of the Mediterranean (PAM) and its Center for Global Studies are committed to preventing global health threats by promoting transboundary cooperation, especially through a One Health approach that links human, animal, and environmental health. In the face of future emergencies, PAM is also committed to promoting greater information sharing for harmonized legislative frameworks and equitable access to resources, to strengthen the resilience of global health systems, especially in developing countries. In both the past and recent history, various outbreaks have been attributed to proven or alleged bioterrorist attacks targeting human or animal populations. This study discusses the general characteristics of several relevant transboundary diseases. Paying high attention to One Health is of utmost importance. However, for a full understanding, it is necessary to consider all related aspects and implications. Full article
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12 pages, 3677 KB  
Article
Identification of Unknown Biological Toxin Proteins Using Mass Spectrometry: A Case Study on De Novo Sequencing of Ricin
by Yubo Song, Hao Wang, Junjie Wen, Jiale Xu, Siyu Zhu, Fuli Wang and Yongqian Zhang
Toxins 2025, 17(11), 564; https://doi.org/10.3390/toxins17110564 - 18 Nov 2025
Viewed by 939
Abstract
Background: The rapid and reliable identification of unknown or highly variable biological toxin proteins, such as the potent Ricin toxin, remains a critical challenge in biodefense and public security. Methods: To address this, we developed a Heuristic De Novo Sequencing (HDPS) strategy, which [...] Read more.
Background: The rapid and reliable identification of unknown or highly variable biological toxin proteins, such as the potent Ricin toxin, remains a critical challenge in biodefense and public security. Methods: To address this, we developed a Heuristic De Novo Sequencing (HDPS) strategy, which combines multiple enzymatic and microwave-assisted acid hydrolysis to generate diverse peptides, followed by a two-stage assembly process integrating de novo sequencing with homology-based database searching for robust error correction. Results: When applied to Ricin, this approach achieved 100% sequence coverage for both its A and B chains, with amino acid-level accuracies of 98.13% and 98.47%, respectively, and successfully corrected potential sequencing ambiguities. Conclusions: These results demonstrate that HDPS is a highly accurate and effective method for the de novo sequencing of full-length proteins, making it particularly valuable for characterizing unknown or mutated toxins in the absence of comprehensive reference databases. Full article
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13 pages, 3687 KB  
Article
Stretchable Porous Membranes for Barrier Tissue Models with Real-Time Measurement and Biomimetic Cyclic Strain
by Alexander P. M. Guttenplan, Joseph W. F. Robertson and Darwin R. Reyes
Micromachines 2025, 16(11), 1282; https://doi.org/10.3390/mi16111282 - 13 Nov 2025
Viewed by 2686
Abstract
In recent years, the development of stretchable electronic devices with mechanical properties similar to those of human tissues has attracted increasing research interest in biomedical engineering, wearables, and other fields. These devices have demonstrated, and some other researchers have already shown, promising advancements [...] Read more.
In recent years, the development of stretchable electronic devices with mechanical properties similar to those of human tissues has attracted increasing research interest in biomedical engineering, wearables, and other fields. These devices have demonstrated, and some other researchers have already shown, promising advancements towards applications that span from measurements of the disruption of model barrier tissues to wearable or implantable devices, soft robotics, and the development of flexible and stretchable batteries. For example, models of barrier tissues, consisting of two compartments separated by a porous membrane, have been used to measure their integrity as well as to investigate the passage of drugs, toxins, and cancer cells through these tissues. Some of these models include an elastomeric membrane which can be stretched to model processes such as breathing and gut peristalsis, while others include electrodes for real-time measurement of barrier tissue integrity. However, to date, microelectrodes have not been fabricated directly on a porous elastomeric membrane. Here, we present lithographically patterned gold electrodes on porous PDMS membranes that enable electronic sensing capabilities in addition to mechanical manipulation. These membranes are incorporated into vacuum-actuated devices which impart cyclic mechanical strain, and their suitability for electrical impedance measurements, even after 1000 stretching cycles under fluids similar to cell culture media, is demonstrated. In the future, we expect to use these electrodes to measure the disruption in model cell barriers as well as to dielectrophoretically trap cells in a region of interest for more rapid assembly of a model tissue. Other areas like wearables, robotics, and power sources will greatly benefit from the further development of this technology. Full article
(This article belongs to the Section E:Engineering and Technology)
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45 pages, 5703 KB  
Review
From Artisan Experience to Scientific Elucidation: Preparation Processes, Microbial Diversity, and Food Applications of Chinese Traditional Fermentation Starters (Qu)
by Dandan Song, Xian Zhong, Yashuai Wu, Jiaqi Guo, Lulu Song and Liang Yang
Foods 2025, 14(22), 3814; https://doi.org/10.3390/foods14223814 - 7 Nov 2025
Cited by 2 | Viewed by 2240
Abstract
Background: Qu was the core starter of traditional Chinese fermentation and had long relied on artisan experience, which led to limited batch stability and standardization. This review organized the preparation processes, microbial diversity, and application patterns of qu in foods from experience to [...] Read more.
Background: Qu was the core starter of traditional Chinese fermentation and had long relied on artisan experience, which led to limited batch stability and standardization. This review organized the preparation processes, microbial diversity, and application patterns of qu in foods from experience to science perspective. Methods: This work summarized typical process parameters for daqu, xiaoqu, hongqu, wheat bran or jiangqu, douchi qu, and qu for mold curd blocks used for furu. Parameters covered raw material moisture, bed thickness, aeration or turning, drying, final moisture, and classification by peak temperature. Multi-omics evidence was used to analyze the coupling of temperature regime, community assembly, and functional differentiation. Indicators for pigment or enzyme production efficiency and safety control such as citrinin in hongqu were included. Results: Daqu showed low, medium, and high temperature regimes. Thermal history governed differences in communities and enzyme profiles and determined downstream fermentation fitness. Xiaoqu rapidly established a three-stage symbiotic network of Rhizopus, Saccharomyces, and lactic acid bacteria, which supported integrated saccharification and alcohol fermentation. Hongqu centered on Monascus and achieved coordinated pigment and aroma formation with toxin risk control through programmed control of temperature, humidity, and final moisture. Wheat bran or jiangqu served as an enzyme production engine for salt-tolerant fermentation, and the combined effects of heat and humidity during the qu period, aeration, and bed loading determined hydrolysis efficiency in salt. Douchi and furu mold curd blocks used thin-layer cultivation and near-saturated humidity to achieve stable mold growth and reproducible interfacial moisture. Conclusions: Parameterizing and online monitoring of key variables in qu making built a process fingerprint with peak temperature, heating rate, and moisture rebound curve at its core. Standardization and functional customization guided by temperature regime, community, and function were the key path for the transition of qu from workshop practice to industry and from experience to science. This approach provided replicable solutions for flavor consistency and safety in alcoholic beverages, sauces, vinegars, and soybean products. Full article
(This article belongs to the Special Issue Sensory Detection and Analysis in Food Industry)
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26 pages, 2960 KB  
Article
Tissue-Specific Gene Expression of Digestive Tract Glands in Paroctopus digueti: Insights for Cephalopod Biology and Aquaculture
by María G. Martínez-Morales, Oscar E. Juárez, Dariel Tovar-Ramírez, Clara E. Galindo-Sánchez, Claudia Ventura-López, Carlos Rosas, Héctor Nolasco-Soria and Bertha Patricia Ceballos-Vázquez
Animals 2025, 15(21), 3224; https://doi.org/10.3390/ani15213224 - 6 Nov 2025
Viewed by 1354
Abstract
Pacific pygmy octopus Paroctopus digueti is a promising model for cephalopod research and aquaculture; its feeding and nutritional biology remain poorly understood. The anterior salivary glands (ASG), posterior salivary glands (PSG), and digestive gland (DG) are central to these processes, but molecular comparisons [...] Read more.
Pacific pygmy octopus Paroctopus digueti is a promising model for cephalopod research and aquaculture; its feeding and nutritional biology remain poorly understood. The anterior salivary glands (ASG), posterior salivary glands (PSG), and digestive gland (DG) are central to these processes, but molecular comparisons are lacking. To address this gap, we performed a transcriptomic study to explore the enzymatic repertoire and functional specialization of these tissues. Total RNA was extracted from ASG, PSG, and DG of three pre-adult individuals collected in La Paz Bay, Mexico. RNA-Seq libraries were sequenced, and a non-redundant multi-tissue transcriptome was assembled. The ASG displayed high expression of neuropeptides, playing a role in neuroendocrine regulation. The PSG showed elevated protease expression, supporting its function in extracellular digestion, alongside toxins that reinforce its role as a venom gland. The DG was enriched in proteins linked to biomolecule catabolism and antimicrobial peptides, alluding to metabolic specialization and immune defense. These results were validated by qPCR, and target genes were also amplified in Octopus maya and O. hubbsorum, showing some similarities in expression patterns. Overall, our findings suggest strong glandular specialization in P. digueti, providing insights into cephalopod digestive physiology and supporting its value as a model species. Full article
(This article belongs to the Special Issue Recent Advances in Cephalopod Biology Research)
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21 pages, 2744 KB  
Article
Genomic Surveillance and Resistance Profiling of Multidrug-Resistant Acinetobacter baumannii Clinical Isolates: Clonal Diversity and Virulence Insights
by Maria Vittoria Ristori, Ilaria Pirona, Lucia De Florio, Sara Elsa Aita, Gabriele Macari, Silvia Spoto, Raffaele Antonelli Incalzi and Silvia Angeletti
Microorganisms 2025, 13(11), 2429; https://doi.org/10.3390/microorganisms13112429 - 23 Oct 2025
Viewed by 1111
Abstract
Acinetobacter baumannii is a multidrug-resistant opportunistic pathogen that poses critical challenges in hospital settings due to its environmental resilience and high resistance to antibiotics. Genomic surveillance has become essential for identifying transmission patterns, guiding antimicrobial stewardship, and informing infection control policies. We conducted [...] Read more.
Acinetobacter baumannii is a multidrug-resistant opportunistic pathogen that poses critical challenges in hospital settings due to its environmental resilience and high resistance to antibiotics. Genomic surveillance has become essential for identifying transmission patterns, guiding antimicrobial stewardship, and informing infection control policies. We conducted whole-genome sequencing on 44 A. baumannii isolates collected between 2022 and 2023 from diverse wards in an Italian hospital. Illumina-based sequencing was followed by a comprehensive bioinformatics pipeline, including genome assembly, taxonomic validation, MLST, SNP-based phylogeny, pan-genome analysis, antimicrobial resistance (AMR) gene profiling, and virulence factor prediction. Most isolates were classified as ST2; SAMPLE-34 was ST1 and genetically distinct. Phylogenetic analysis revealed four clonal clusters with cluster-specific AMR and accessory gene content. The pan-genome included 5050 genes, with notable variation linked to hospital ward origin. ICU and internal medicine strains carried higher loads of AMR genes, especially against aminoglycosides, β-lactams, and quinolones. Virulence profiling highlighted widespread immune evasion mechanisms; “Acenovactin” was predominant, while some isolates lacked key adhesion or toxin factors. Our findings underscore the clinical relevance of integrating genomic epidemiology into routine hospital surveillance. Identifying clonal clusters and resistance signatures supports real-time outbreak detection, risk stratification, and targeted infection prevention strategies. Full article
(This article belongs to the Collection Feature Papers in Antimicrobial Agents and Resistance)
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20 pages, 1024 KB  
Review
Chromosomal Roadblocks in Male Fertility: Mechanisms, Risk Factors and Syndromes
by Achilleas G. Mitrakas, Christina-Angelika Alexiadi, Sofia Gargani, Triantafyllos Alexiadis, Sofia-Panagiota Alexopoulou, Olga Pagonopoulou and Maria Lambropoulou
Medicina 2025, 61(10), 1864; https://doi.org/10.3390/medicina61101864 - 16 Oct 2025
Cited by 1 | Viewed by 2975
Abstract
Male infertility affects nearly 15% of couples worldwide, with chromosomal abnormalities representing a major underlying cause. This review explores how numerical and structural chromosomal anomalies, along with environmental exposures, lifestyle factors, and age-related genetic changes, disrupt spermatogenesis and contribute to infertility. It synthesizes [...] Read more.
Male infertility affects nearly 15% of couples worldwide, with chromosomal abnormalities representing a major underlying cause. This review explores how numerical and structural chromosomal anomalies, along with environmental exposures, lifestyle factors, and age-related genetic changes, disrupt spermatogenesis and contribute to infertility. It synthesizes findings from cytogenetic, molecular, and clinical studies, with particular focus on mechanisms such as meiotic nondisjunction, spindle assembly checkpoint dysfunction, and alterations in cohesin and synaptonemal complex proteins. Chromosomal abnormalities, both numerical and structural, emerge as key contributors to male infertility by impairing chromosomal segregation and recombination, often leading to azoospermia or oligospermia. Meiotic checkpoint failures and recombination errors further exacerbate the production of aneuploid sperm. Environmental toxins, oxidative stress, and poor nutrition disrupt hormonal balance and chromatin integrity, while advancing paternal age is associated with increased sperm aneuploidy and impaired meiotic control, with implications for assisted reproduction. Specific syndromes, including AZF deletions, Kallmann syndrome, and 46,XX testicular DSD, exemplify the direct genetic impact on male fertility. Overall, chromosomal abnormalities are central to the pathophysiology of male infertility, arising from intrinsic meiotic errors as well as extrinsic environmental and lifestyle factors. Integrating cytogenetic diagnostics, genetic counseling, and lifestyle interventions is essential for comprehensive fertility assessment and management. Further research into molecular biomarkers and targeted therapies could enhance diagnosis, improve treatment strategies, and lead to better reproductive outcomes. Full article
(This article belongs to the Special Issue From Conception to Birth: Embryonic Development and Disease)
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18 pages, 2398 KB  
Article
Genome Analysis of Alternaria alstroemeriae L6 Associated with Black Spot of Strawberry: Secondary Metabolite Biosynthesis and Virulence
by Li Zhang, Boyuan Zhang, Lizhu Shao, Miaomiao Yang, Xueling Zhao, Ziyu Wang, Yingjun Zhang, Yuting Li, Yating Wang, Yuansen Hu and Peng Li
J. Fungi 2025, 11(10), 710; https://doi.org/10.3390/jof11100710 - 30 Sep 2025
Viewed by 1060
Abstract
A pathogenic fungus was isolated from the leaves of strawberry black spot in Zhengzhou China. Based on morphological and phylogenetic analysis, the isolate was identified as Alternaria alstroemeriae. Hybrid sequencing and assembly yielded a high-quality 38.7 Mb genome with 12,781 predicted genes [...] Read more.
A pathogenic fungus was isolated from the leaves of strawberry black spot in Zhengzhou China. Based on morphological and phylogenetic analysis, the isolate was identified as Alternaria alstroemeriae. Hybrid sequencing and assembly yielded a high-quality 38.7 Mb genome with 12,781 predicted genes and 99.6% Benchmarking Universal Single-Copy Orthologs (BUSCO) completeness. Functional annotation revealed enrichment in carbohydrate metabolism, secondary metabolite biosynthesis, and virulence-associated genes. Strain L6 harbored 45 biosynthetic gene clusters(BGCs), including 12 clusters for terpenes, 7 for non-ribosomal peptide synthetases, and 7 for polyketide synthases. Six BGCs showed high similarity to known pathways producing alternariol (phytotoxic/mycotoxic compound), alternapyrone (phytotoxin), choline (osmoprotectant), terpestacin (anti-angiogenic agent), clavaric acid (anticancer terpenoid), and betaenone derivatives (phytotoxins). CAZyme analysis identified 596 carbohydrate-active enzymes, aligning with L6’s biotrophic lifestyle. Additionally, 996 secreted proteins were predicted, of which five candidate effectors contained the conserved RxLx [EDQ] host-targeting motif, suggesting potential roles in virulence. This genome resource highlights L6’s exceptional secondary metabolites (SMs) diversity, featuring both plant-pathogenic toxins and pharmacologically valuable compounds, indicating that this endophytic fungus is a potential producer of metabolites meriting further exploration and development. Full article
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19 pages, 6870 KB  
Article
Genomic Markers Distinguishing Shiga Toxin-Producing Escherichia coli: Insights from Pangenome and Phylogenomic Analyses
by Asmaa Elrefaey, Kingsley E. Bentum, Emmanuel Kuufire, Tyric James, Rejoice Nyarku, Viona Osei, Yilkal Woube, Temesgen Samuel and Woubit Abebe
Pathogens 2025, 14(9), 862; https://doi.org/10.3390/pathogens14090862 - 30 Aug 2025
Viewed by 1609
Abstract
Shiga toxin-producing Escherichia coli (STEC) are genetically diverse foodborne pathogens of major global public health concerns. Serogroup-level identification is critical for effective surveillance and outbreak control; however, it is often challenged by STEC’s genome plasticity and frequent recombination. In this study, we employed [...] Read more.
Shiga toxin-producing Escherichia coli (STEC) are genetically diverse foodborne pathogens of major global public health concerns. Serogroup-level identification is critical for effective surveillance and outbreak control; however, it is often challenged by STEC’s genome plasticity and frequent recombination. In this study, we employed a standardized pangenomic pipeline integrating Roary ILP Bacterial Core Annotation Pipeline (RIBAP) and Panaroo to analyze 160 complete, high-quality STEC genomes representing eight major serogroups at a 95% sequence identity threshold. Candidate serogroup-specific markers were identified using gene presence/absence profiles from RIBAP and Panaroo. Our analysis revealed several high-confidence markers, including metabolic genes (dgcE, fcl_2, dmsA, hisC) and surface polysaccharide-related genes (capD, rfbX, wzzB). Comparative pangenomic evaluation showed that RIBAP predicted a larger pangenome size than Panaroo. Additionally, some genomes from the O104:H1, O145:H28, and O45:H2 serotypes clustered outside their expected clades, indicating sporadic serotype misplacements in phylogenetic reconstructions. Functional annotation suggested that most candidate markers are involved in critical processes such as glucose metabolism, lipopolysaccharide biosynthesis, and cell surface assembly. Notably, approximately 22.9% of the identified proteins were annotated as hypothetical. Overall, this study highlights the utility of pangenomic analysis for potential identification of clinically relevant STEC serogroups markers and phylogenetic interpretation. We also note that pangenome analysis could guide the development of more accurate diagnostic and surveillance tools. Full article
(This article belongs to the Section Bacterial Pathogens)
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29 pages, 4778 KB  
Article
In Silico Development of a Chimeric Multi-Epitope Vaccine Targeting Helcococcus kunzii: Coupling Subtractive Proteomics and Reverse Vaccinology for Vaccine Target Discovery
by Khaled S. Allemailem
Pharmaceuticals 2025, 18(9), 1258; https://doi.org/10.3390/ph18091258 - 25 Aug 2025
Cited by 4 | Viewed by 2523
Abstract
Background: Helcococcus kunzii, a facultative anaerobe and Gram-positive coccus, has been documented as a cunning pathogen, mainly in immunocompromised individuals, as evidenced by recent clinical and microbiological reports. It has been associated with a variety of polymicrobial infections, comprising diabetic foot [...] Read more.
Background: Helcococcus kunzii, a facultative anaerobe and Gram-positive coccus, has been documented as a cunning pathogen, mainly in immunocompromised individuals, as evidenced by recent clinical and microbiological reports. It has been associated with a variety of polymicrobial infections, comprising diabetic foot ulcers, prosthetic joint infections, osteomyelitis, endocarditis, and bloodstream infections. Despite its emerging clinical relevance, no licensed vaccine or targeted immunotherapy currently exists for H. kunzii, and its rising resistance to conventional antibiotics presents a growing public health concern. Objectives: In this study, we employed an integrated subtractive proteomics and immunoinformatics pipeline to design a multi-epitope subunit vaccine (MEV) candidate against H. kunzii. Initially, pan-proteome analysis identified non-redundant, essential, non-homologous, and virulent proteins suitable for therapeutic targeting. Methods/Results: From these, two highly conserved and surface-accessible proteins, cell division protein FtsZ and peptidoglycan glycosyltransferase FtsW, were selected as promising vaccine targets. Comprehensive epitope prediction identified nine cytotoxic T-lymphocyte (CTL), five helper T-lymphocyte (HTL), and two linear B-cell (LBL) epitopes, which were rationally assembled into a 397-amino-acid-long chimeric construct. The construct was designed using appropriate linkers and adjuvanted with the cholera toxin B (CTB) subunit (NCBI accession: AND74811.1) to enhance immunogenicity. Molecular docking and dynamics simulations revealed persistent and high-affinity ties amongst the MEV and essential immune receptors, indicating a durable ability to elicit an immune reaction. In silico immune dynamic simulations predicted vigorous B- and T-cell-mediated immune responses. Codon optimization and computer-aided cloning into the E. coli K12 host employing the pET-28a(+) vector suggested high translational efficiency and suitability for bacterial expression. Conclusions: Overall, this computationally designed MEV demonstrates favorable immunological and physicochemical properties, and presents a durable candidate for subsequent in vitro and in vivo validation against H. kunzii-associated infections. Full article
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21 pages, 6129 KB  
Article
Diversity and Novelty of Venom Peptides in Vermivorous Cone Snails, Subgenus Rhizoconus (Gastropoda: Mollusca)
by Christine Marie C. Florece, Quentin Kaas, Neda Barghi and Arturo O. Lluisma
Mar. Drugs 2025, 23(7), 266; https://doi.org/10.3390/md23070266 - 26 Jun 2025
Viewed by 2420
Abstract
A large majority of cone snails (a species in the genus Conus) are vermivorous (worm-hunting), but the diversity and bioactivity of their venom peptides remain largely unexplored. In this study, we report the first venom gland transcriptomes from two species in the [...] Read more.
A large majority of cone snails (a species in the genus Conus) are vermivorous (worm-hunting), but the diversity and bioactivity of their venom peptides remain largely unexplored. In this study, we report the first venom gland transcriptomes from two species in the Rhizoconus clade, Conus capitaneus and Conus mustelinus, and a new Conus miles transcriptome from a specimen collected in the Philippines. From the set of assembled sequences, a total of 225 C. capitaneus, 121 C. miles, and 168 C. mustelinus putative peptide toxin transcripts were identified, which were assigned to 27 canonical gene superfamilies in C. capitaneus and 24 in C. miles and in C. mustelinus. Most of these venom peptides are novel, and some exhibit new cysteine patterns. Clustering also revealed 12 putative novel gene superfamilies, highlighting the diversity of uncharacterized venom peptides in this group. The O1-, M-, O2-, and con-ikot-ikot superfamilies were the most abundant, while gene superfamilies such as D and G2 were highly expressed. Several hormone-like conopeptides were also identified in this study, revealing the vast diversity of conopeptides from the Rhizoconus species. Full article
(This article belongs to the Section Marine Toxins)
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13 pages, 3936 KB  
Article
A Highly Sensitive Silicon-Core Quantum Dot Fluorescent Probe for Vomitoxin Detection in Cereals
by Caiwen Dong, Yaqin Li, Xincheng Sun, Xuehao Yang and Tao Wei
Foods 2025, 14(9), 1545; https://doi.org/10.3390/foods14091545 - 28 Apr 2025
Cited by 2 | Viewed by 1335
Abstract
Vomitoxin is a member of the monotrichous mycotoxin family with a complex chemical structure and significant biological activity. This toxin has strong immunosuppressive toxic effects and can cause serious damage to human and animal health. In this study, an on-site immune detection method [...] Read more.
Vomitoxin is a member of the monotrichous mycotoxin family with a complex chemical structure and significant biological activity. This toxin has strong immunosuppressive toxic effects and can cause serious damage to human and animal health. In this study, an on-site immune detection method based on an immune SiO2@QD fluorescent probe was developed, which realized the rapid and quantitative detection of emetic toxins in grains. Polyethyleneimine (PEI) is a polymer containing a large number of amino groups, and the binding of PEI to the surface of quantum dots can serve to regulate growth and provide functionalized groups. A SiO2@QD nanotag with good dispersibility and a high fluorescence intensity was synthesized by combining a PEI interlayer on the surface of SiO2 nanospheres. Utilizing the electrostatic adsorption of the amino group in PEI, CdSe/ZnS QDs were self-assembled on the surface of SiO2 nanospheres. In the stability test, the SiO2@QDs could maintain basically the same fluorescence intensity for 90 consecutive days in the dark at 4 °C, showing a high fluorescence stability. The fluorescence-enhanced QD immune probe was formed by coupling with anti-DON monoclonal antibodies through carbodiimide chemical synthesis. For the detection of spiked wheat flour samples, the immuno-SiO2@QD fluorescent probe showed excellent sensitivity and stability, the detection limit reached 0.25 ng/mL, and the average recovery rate was 92.2–101.6%. At the same time, the immuno-SiO2@QD fluorescent probe is simple to operate, is capable of rapid responses, and has great potential in the rapid detection of vomitoxins in grains. Full article
(This article belongs to the Section Food Toxicology)
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