Toxins

14 pages, 2345 KB

Open AccessArticle

Development of a UHPLC-MS/MS Method for Quantitative Analysis of Aflatoxin B₁ in Scutellaria baicalensis

by Yuanfang Liu, Cuiping Zeng, Ying-Ying Li, Jiayu Guo and Jinming Xu

Toxins 2025, 17(9), 473; https://doi.org/10.3390/toxins17090473 - 21 Sep 2025

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Aflatoxin B₁ (AFB₁) contamination in Scutellaria baicalensis poses a serious threat to the safety of traditional Chinese medicinal products. In this study, a sensitive and reliable ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination [...] Read more.

Aflatoxin B₁ (AFB₁) contamination in Scutellaria baicalensis poses a serious threat to the safety of traditional Chinese medicinal products. In this study, a sensitive and reliable ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination of AFB₁ in Scutellaria baicalensis. Method optimization included selection of chromatographic columns, mobile phase composition, and mass spectrometric parameters. Sample pretreatment was also optimized to reduce matrix interference and enhance extraction efficiency. The method showed excellent linearity (R² > 0.999) in the range of 0.1–10.0 µg/L, with a limit of detection (LOD) of 0.03 µg/kg and a limit of quantification (LOQ) of 0.10 µg/kg. Precision and recovery studies demonstrated good repeatability and accuracy, with intra- and inter-day relative standard deviations (RSDs) below 5.2% and recoveries ranging from 88.7% to 103.4%. Application of the method to six commercial Scutellaria baicalensis samples revealed detectable AFB₁ in two samples, though all levels were below national safety limits. This method provides a robust tool for routine monitoring of AFB₁ in herbal medicines and supports the establishment of quality control systems for Scutellaria baicalensis. Full article

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13 pages, 1187 KB

Open AccessArticle

Phenanthrene Amplifies Microcystin-Induced Toxicity in the Submerged Macrophyte Vallisneria natans

by Xiang Wan, Yi Zhang, Yucong Li, Fei Yang and Liqiang Xie

Toxins 2025, 17(9), 472; https://doi.org/10.3390/toxins17090472 - 21 Sep 2025

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Abstract

Microcystin–LR (MC-LR) and phenanthrene (Phen), which commonly co-occur in eutrophic waters, have been extensively studied as individual contaminants, but their combined ecotoxicological effects on submerged macrophytes remain unclear. In this study, we examined the individual and combined toxicity of MC-LR (2, 10, 50, [...] Read more.

Microcystin–LR (MC-LR) and phenanthrene (Phen), which commonly co-occur in eutrophic waters, have been extensively studied as individual contaminants, but their combined ecotoxicological effects on submerged macrophytes remain unclear. In this study, we examined the individual and combined toxicity of MC-LR (2, 10, 50, 250, and 1000 μg/L) and Phen (0.2, 1, 5, 25, and 100 μg/L) on the submerged macrophyte Vallisneria natans over a 7-day exposure. Key toxicity biomarkers, including growth, photosynthetic efficiency, and antioxidant responses (catalase, superoxide dismutase, glutathione S-transferase, and malondialdehyde), were evaluated. The results showed that high concentrations of each contaminant alone (MC-LR ≥ 1000 μg/L; Phen ≥ 100 μg/L) significantly inhibited growth and reduced photosynthetic efficiency. In contrast, synergistic toxicity was observed at much lower combined concentrations (≥50 + 5 μg/L), with effects substantially exceeding those of individual exposures. Co-exposure intensified antioxidant activity, but it was insufficient to mitigate oxidative damage. Notably, Phen at concentrations above 25 μg/L significantly enhanced the bioaccumulation of MC-LR in V. natans. These findings demonstrate that environmentally relevant mixtures of MC-LR and Phen induce remarkable toxicity even at concentrations where individual compounds show negligible effects. The results highlight that co-existing cyanotoxins and polycyclic aromatic hydrocarbons may present greater ecological risks than predicted from single-contaminant assessments, underscoring the need to update current ecological risk frameworks for the accurate evaluation of complex pollution scenarios in freshwater systems. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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13 pages, 277 KB

Open AccessArticle

Regulation of Pseudomonas sp. PSC001 on the Artificial Rumen Environment Contaminated by Zearalenone

by Yiming Han, Xinfeng Li, Xiaoli Ren, Chao Song, Zhaojie Zhang, Yufeng Gao, Dongmei Shi, Hongyu Deng, Heping Huangfu and Jinming Wang

Toxins 2025, 17(9), 471; https://doi.org/10.3390/toxins17090471 - 21 Sep 2025

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Abstract

In this study, the RUSITEC system was used to study the regulation of rumen-derived Pseudomonas sp. PSC001 (PSC001) on the rumen environment contaminated by Zearalenone (ZEN). The rumen fluid of dairy cows was selected as the fermentation broth, and four experimental groups were [...] Read more.

In this study, the RUSITEC system was used to study the regulation of rumen-derived Pseudomonas sp. PSC001 (PSC001) on the rumen environment contaminated by Zearalenone (ZEN). The rumen fluid of dairy cows was selected as the fermentation broth, and four experimental groups were set up: control group (CON), Pseudomonas group (PS), ZEN pollution group (ZEN), and PS and ZEN co-treatment group (PS + ZEN). The NH₃-N, microbial protein (MCP), and volatile fatty acid (VFA) in the rumen fermentation broth were measured after culturing, and the changes in microbial community structure in rumen fluid were analyzed by 16S rRNA gene sequencing. After adding PSC001, the concentration of propionic acid, valeric acid, and butyric acid increased, and the acetate to propionate ratio and concentration of isovaleric acid decreased. ZEN exposure can lead to an abnormal increase in NH₃-N, valeric acid, and isovaleric acid content and a decrease in MCP content. The content of NH₃-N, valeric acid, and isovaleric acid decreased and the content of MCP increased in the PS + ZEN combined treatment group. The addition of PSC001 and ZEN significantly or extremely significantly increased the abundance of 18 genera and significantly or extremely significantly decreased the relative abundance of 5 genera in rumen fluid, respectively. It is worth noting that with the addition of both at the same time, the abundance of four genera in the PS + ZEN group was significantly or extremely significantly increased among the five genera with decreased abundance in the ZEN group. Among the 18 genera with increased abundance in the ZEN group, 10 genera in the PS + ZEN group decreased significantly or extremely significantly. In summary, the addition of PSC001 alleviated the negative impact of ZEN on the internal environment of rumen fermentation, and it also had a positive regulatory effect on rumen fermentation. Full article

15 pages, 4348 KB

Open AccessArticle

Macula Densa Alleviates Shiga Toxin-Induced Acute Kidney Injury via CCN1-Mediated Renal Tubular Repair

by Hongzhi Wan, Yuhui Wang, Jiahui Chen, Hongqi Liu, Jiamei Li, Qisheng Su, Hui Peng, Xiaotao Duan and Bo Wang

Toxins 2025, 17(9), 470; https://doi.org/10.3390/toxins17090470 - 21 Sep 2025

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Abstract

Shiga toxins (Stx), produced by Shiga toxin-producing Escherichia coli, preferentially attack renal tissue and frequently induce acute kidney injury (AKI) and renal failure. To prevent irreversible damage, the injured renal tissue, particularly renal tubular epithelium, mounts a remodeling and regeneration response to [...] Read more.

Shiga toxins (Stx), produced by Shiga toxin-producing Escherichia coli, preferentially attack renal tissue and frequently induce acute kidney injury (AKI) and renal failure. To prevent irreversible damage, the injured renal tissue, particularly renal tubular epithelium, mounts a remodeling and regeneration response to repair itself. However, how such intrinsic renal repair processes are initiated and coordinated in infected renal tubular regions remains elusive. Herein, we reported that macula densa apparatus, in addition to its conventional role as a salt sensor in nephron, can function as an endogenous sensor for exogenous toxins (e.g., Stx). We demonstrated that macula densa cells orchestrate a rapid repair niche by initiating transcriptional activation of repair and regeneration factors in both Stx-injured murine models and human kidney organoids. Mechanistically, we showed that in response to Stx exposure, macula densa cells release a specific repair factor CCN1, which effectively promotes the regeneration of toxin-injured renal tubular epithelium and facilitates renal tubular repair through integrin-mediated signaling pathways. Moreover, we demonstrated that treatment with recombinant CCN1 can greatly ameliorate the structural damage and significantly restore the proximal tubular reabsorption capacity in Stx-infected kidney organoids. Our finding highlights a novel role of macula densa apparatus in toxin-induced renal injury, and paves a new avenue for treatment of AKI-associated renal diseases. Full article

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39 pages, 1701 KB

Open AccessReview

Bioorganic Chemistry, Toxinology, and Pharmaceutical Uses of Datura Metabolites and Derivatives

by Amin Mahmood Thawabteh, Saleh Sulaiman, Ilaf Omar Alabed, Laura Scrano, Donia Karaman, Rafik Karaman and Sabino A. Bufo

Toxins 2025, 17(9), 469; https://doi.org/10.3390/toxins17090469 - 18 Sep 2025

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Abstract

Datura species have been recognized for their potent pharmacological properties, producing a diverse array of tropane and non-tropane alkaloids with significant clinical and toxicological relevance. This review synthesizes current knowledge on the biosynthesis, pharmacology, and therapeutic applications of 43 compounds isolated from Datura [...] Read more.

Datura species have been recognized for their potent pharmacological properties, producing a diverse array of tropane and non-tropane alkaloids with significant clinical and toxicological relevance. This review synthesizes current knowledge on the biosynthesis, pharmacology, and therapeutic applications of 43 compounds isolated from Datura, with emphasis on both major constituents—such as atropine, hyoscyamine, and scopolamine—and minor alkaloids, including anisodamine, apoatropine, and datumetine. These alkaloids were classified into four significant categories, drawing on recent advances in plant biochemistry and analytical chemistry. The analysis is based on 204 peer-reviewed scientific publications from the past decade (2015–2025), highlighting both traditional ethnobotanical knowledge and recent pharmacological advances. The review details their enzymatic pathways, mechanisms of action at muscarinic and other receptor systems, pharmacokinetics, and dose-dependent toxicological profiles. Particular attention is given to lesser-studied derivatives and metabolites with emerging therapeutic potential, as well as their role in metabolic engineering, drug discovery, and forensic analysis. Notably, datum tine is highlighted for its unique NMDA receptor modulatory effects and neurotoxic potential, while tropine and hygrine serve as critical biosynthetic intermediates and analytical markers. By integrating biochemical, pharmacological, and toxicological insights, this work provides a comprehensive framework for future exploration of Datura alkaloids as both therapeutic agents and research tools. Full article

(This article belongs to the Section Plant Toxins)

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21 pages, 1952 KB

Open AccessArticle

Integrated Assessment of Fungi Contamination and Mycotoxins Levels Across the Rice Processing Chain

by Carolina Sousa Monteiro, Eugénia Pinto, Rosalía López-Ruiz, Jesús Marín-Sáez, Antonia Garrido Frenich, Miguel A. Faria and Sara C. Cunha

Toxins 2025, 17(9), 468; https://doi.org/10.3390/toxins17090468 - 18 Sep 2025

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Abstract

This study investigated the occurrence of fungi and mycotoxins throughout the rice processing chain, from paddy rice to final white rice, in two rice varieties (variety I and variety II). A total of 75 fungal isolates were identified, belonging to the genera Penicillium [...] Read more.

This study investigated the occurrence of fungi and mycotoxins throughout the rice processing chain, from paddy rice to final white rice, in two rice varieties (variety I and variety II). A total of 75 fungal isolates were identified, belonging to the genera Penicillium, Alternaria, Aspergillus, Fusarium, and Talaromyces. Variety I exhibited a higher prevalence of Penicillium and Alternaria, whereas Variety II was dominated mainly by Alternaria, accounting for 63% of all isolates. Multi-mycotoxin screening of 22 mycotoxins revealed contamination by tenuazonic acid (TeA), zearalenone (ZEN), and 15-acetyl-deoxynivalenol (15-AcDON), with TeA concentrations exceeding 4000 µg/kg in whitened rice of variety II. Cluster analysis showed paddy and brown rice grouping together due to higher fungal loads and toxin levels, whereas whitened and final white rice clustered separately, reflecting reduced fungal counts but persistence of TeA, 15-AcDON, ZEN, and citrinin (CIT). The co-clustering of Alternaria with TeA and ZEN indicates strong field-related contamination. Although processing significantly decreased fungal loads, residual toxins persisted, emphasizing that rice polishing does not fully mitigate mycotoxin risks. These findings underscore the need for comprehensive surveillance and integrated management practices across the rice supply chain to minimize potential health hazards associated with fungal contaminants and their toxic metabolites. Full article

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19 pages, 3634 KB

Open AccessArticle

Chlorogenic Acid and VX765 Alleviate Deoxynivalenol-Induced Enterohepatic Injury and Lipid Metabolism Disorders by Improving Intestinal Microecology

by Tao Wen, Sirui Li, Huijun Guo, Jinbo Wang, Xinru Zhang and Chunyang Wang

Toxins 2025, 17(9), 467; https://doi.org/10.3390/toxins17090467 - 18 Sep 2025

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Abstract

Widespread contamination of deoxynivalenol (DON) in cereals and feed threatens global food safety. This study investigated the effects of Chlorogenic acid (CGA) and VX765 on DON-induced enterohepatic injury. A total of 48 female mice were divided into four groups: control (normal saline), DON [...] Read more.

Widespread contamination of deoxynivalenol (DON) in cereals and feed threatens global food safety. This study investigated the effects of Chlorogenic acid (CGA) and VX765 on DON-induced enterohepatic injury. A total of 48 female mice were divided into four groups: control (normal saline), DON (1 mg/kg.bw), CGA (100 mg/kg.bw CGA + 1 mg/kg.bw DON), and VX765 (100 mg/kg.bw VX765 + 1 mg/kg.bw DON). After 28-day gavage period, the results showed that CGA and VX765 reduced DON-induced intestinal barrier damage. Metabolomics data revealed that CGA and VX765 restored cecal microbiota structure and alleviated DON-induced hepatic injury and lipid metabolic disorders by reshaping intestinal microbiota. Retrograde endocannabinoid signaling was identified as a critical pathway for cecal microbial metabolism and hepatic lipid regulation mediated by CGA and VX765. Additionally, CGA and VX765 reversed the upregulation of IMPA, CDS2, DGKA, NDUFS8, and MAPK1 mRNA and protein expression levels induced by DON via the microbiota-gut-liver axis. Full article

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15 pages, 605 KB

Open AccessArticle

Metabolic and Phytotoxic Profile of Phytopathogens in Main Extensive Crops of Argentina

by Francisco José Sautua, Maria Chiara Zonno, Pierluigi Reveglia, Maria Letizia Ciavatta, Marianna Carbone, Lucia Lecce, María Cecilia Pérez-Pizá, Gaetano Corso, Marcelo Anibal Carmona and Antonio Evidente

Toxins 2025, 17(9), 466; https://doi.org/10.3390/toxins17090466 - 18 Sep 2025

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Abstract

Phytopathogenic fungi represent a significant biotic stress affecting global agriculture, often causing severe diseases and, in some cases, leading to plant death. They have been isolated from economically important crops, including cereals, legumes, and fruits. Among the compounds produced by fungi, phytotoxins play [...] Read more.

Phytopathogenic fungi represent a significant biotic stress affecting global agriculture, often causing severe diseases and, in some cases, leading to plant death. They have been isolated from economically important crops, including cereals, legumes, and fruits. Among the compounds produced by fungi, phytotoxins play a key role in disease development by interfering with host physiological processes. In this study, organic extracts from Cercospora kikuchii, Cercospora nicotianae, Cercospora sojina, Diaporthe longicolla, Septoria glycines, Pyrenophora teres, and Pyrenophora tritici-repentis, isolated from three major Argentine crops, were first screened for the in vitro production of phytotoxic metabolites. Subsequently, selected metabolites were dereplicated using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The phytotoxins identified varied according to the fungal species and extraction conditions. Cercosporin, putaminoxin, scytalone, and isosclerone were identified. These findings underscore the need for further chemical investigation to comprehensively characterize the metabolome of these phytopathogens and clarify their roles in plant–pathogen interactions. Full article

(This article belongs to the Topic Application of Analytical Technology in Metabolomics)

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14 pages, 3398 KB

Open AccessArticle

Prokaryotic and Eukaryotic Community Succession and Potential Parasitic Interactions During Two Alexandrium pacificum Blooms in Aotearoa New Zealand

by Laura Biessy, Lincoln Mackenzie and Kirsty F. Smith

Toxins 2025, 17(9), 465; https://doi.org/10.3390/toxins17090465 - 17 Sep 2025

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Abstract

Harmful algal blooms (HABs), caused by the dinoflagellate Alexandrium pacificum, are increasingly frequent in the Marlborough Sounds, an important aquaculture region in Aotearoa New Zealand. Alexandrium pacificum produces paralytic shellfish toxins and blooms cause significant economic and ecological disruptions through contamination of [...] Read more.

Harmful algal blooms (HABs), caused by the dinoflagellate Alexandrium pacificum, are increasingly frequent in the Marlborough Sounds, an important aquaculture region in Aotearoa New Zealand. Alexandrium pacificum produces paralytic shellfish toxins and blooms cause significant economic and ecological disruptions through contamination of edible shellfish. High-throughput sequencing of prokaryotic and eukaryotic communities was used to investigate community dynamics during bloom events across two consecutive summers. Distinct successional shifts were observed, with prokaryotic communities dominated by Rhodobacterales and Flavobacteriales during blooms, and increased abundance of the SAR11 clade (Pelagibacterales) post-bloom. Eukaryotic diversity was dominated by Alexandrium species (Gonyaulacales) during the bloom, and subsequently shifted towards Syndiniales, Gymnodiniales, and Peridiniales as blooms collapsed. Significant correlations indicated potential ecological roles for these taxa in bloom regulation, particularly Syndiniales, which could indicate parasitic interactions. Depth profiles revealed consistent microbial composition throughout the water column, validating depth-integrated sampling strategies for community studies. This research describes changes in the composition of microbial communities during two A. pacificum blooms, suggesting that species interactions (e.g., via parasitism) may play a role shaping bloom dynamics. Further studies incorporating environmental parameters, especially nutrient dynamics linked to anthropogenic activities, are necessary to better understand the drivers of blooms in this important aquaculture region. Full article

(This article belongs to the Special Issue Harmful Algae in a Changing World: Where Did We Come from and Where Are We Going?)

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23 pages, 4900 KB

Open AccessArticle

Biodiversity-Driven Screening of Amphibian Skin Secretions for Inflammatory Modulation in Joint Diseases

by Douglas Souza Oliveira, César Alexandre, Miryam Paola Alvarez-Flores, Isadora Maria Villas-Boas, Hugo Vigerelli, Isabel de Fátima Correia Batista, Michelle Cristiane Bufalo, Nancy Starobinas, Flávio Lichtenstein, Rafael Marques-Porto, Marcus Buri, Viviane Portas-Lopes, Pedro Luiz Mailho-Fontana, Marta Maria Antoniazzi, Denise Vilarinho Tambourgi, Ana Marisa Chudzinski-Tavassi, Catarina Teixeira, Carlos Jared and Olga Martinez Ibañez

Toxins 2025, 17(9), 464; https://doi.org/10.3390/toxins17090464 - 17 Sep 2025

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Abstract

This study explores the direct effects of amphibian skin secretions on human cells involved in joint diseases, aiming to identify species with potential for inflammatory modulation. Secretions were obtained from sixteen species distributed across Brazilian biomes and one European species. Following biochemical characterization, [...] Read more.

This study explores the direct effects of amphibian skin secretions on human cells involved in joint diseases, aiming to identify species with potential for inflammatory modulation. Secretions were obtained from sixteen species distributed across Brazilian biomes and one European species. Following biochemical characterization, human chondrocytes, synoviocytes, and macrophages were treated with secretions for 24 h. The cytotoxicity and modulation of the IL-6, IL-8, TNF-α, and IL-1β release were assessed. Synoviocytes showed the greatest resistance to cytotoxic effects, though sensitivity varied by species. Secretions from Trachycephalus mesophaeus, Pipa carvalhoi, and Phyllomedusa bahiana exhibited the highest cytotoxicity. At non-cytotoxic concentrations, P. carvalhoi and Leptodactylus fuscus strongly induced IL-6 and IL-8 in chondrocytes and synoviocytes, with P. carvalhoi also stimulating IL-1β and TNF-α release in macrophages. Among Bufonidae species, particularly Rhinella jimi and Bufo bufo, were potent inducers of TNF-α and IL-1β in macrophages. Secretions lacking pro-inflammatory effects were further tested for anti-inflammatory activity. P. bahiana reduced TNF-α production in stimulated macrophages and IL-6 in synoviocytes, while Siphonops annulatus and T. mesophaeus reduced LPS-induced TNF-α in macrophages. Our data underscore the rich biodiversity of amphibians, supporting the bioprospecting of their cutaneous secretions. These data reveal substantial potential for uncovering bioactive compounds with pharmacological applications. Full article

(This article belongs to the Special Issue Venoms and Drugs)

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13 pages, 1493 KB

Open AccessArticle

OnabotulinumtoxinA Is an Effective Treatment for Reducing the Interictal Burden in Patients with Chronic Migraine: A Prospective Observational Study

by Alejandro Sánchez-Huertas, Oscar Camejo-Mas, Sebastian Garcia-Roldan, Rocio Alonso-Castillo, Lara Pulido-Fraiz, Andrea Higuera Ruiz de la Hermosa, Leonardo Portocarrero-Sánchez and Javier Díaz-de-Terán

Toxins 2025, 17(9), 463; https://doi.org/10.3390/toxins17090463 - 16 Sep 2025

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Abstract

Interictal burden (IB), defined as the symptoms and impairments that occur between migraine attacks, including cognitive dysfunction, photophobia, and fatigue, is recognized as a significant determinant of quality of life in patients. A prospective observational study was conducted. Patients diagnosed with chronic migraine [...] Read more.

Interictal burden (IB), defined as the symptoms and impairments that occur between migraine attacks, including cognitive dysfunction, photophobia, and fatigue, is recognized as a significant determinant of quality of life in patients. A prospective observational study was conducted. Patients diagnosed with chronic migraine (CM) and under treatment with OnabotulinumtoxinA (OnabotA) according to the PREEMPT protocol (every 12 weeks) were assessed at baseline and at 3, 6, 9, and 12 months. The primary endpoint was to evaluate the change in the IB measured with the Migraine Interictal Burden Scale (MIBS-4) and in the monthly migraine days (MMD). The secondary endpoint was acute medication use. This single-center study included 150 patients (91.3% female; median age 44 years). MIBS-4 scores were decreased by 29.1% at 3 months (8.47 to 5.97) and by 41.6% at 12 months (to 4.86; p < 0.001). IB-free status was achieved by 16 patients (10.7%). The most disabling baseline symptoms were photophobia (37%), fatigue (20%), and allodynia (18%), which reduced by 52%, 43%, and 39% at 12 months, respectively. MMD were reduced from 18.6 to 8.3 days at 12 months and triptan and analgesic intake decreased by 58.7% and 55.4%. OnabotA significantly reduced both IB and migraine frequency over 12 months, underscoring its relevance in CM management. Full article

(This article belongs to the Special Issue Botulinum Neurotoxins for the Treatment of Chronic Pain and Headaches)

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17 pages, 6509 KB

Open AccessArticle

Cost-Effective and Rapid Detection of Tetrodotoxin Using Indium Tin Oxide Electrodes via In Vitro Electrophysiology and Electrochemistry

by Naga Adithya Chandra Pandurangi, Manel M. Santafe, Angels Tudo, Nagihan Ozsoy, Fransesc X. Sureda, Mark L. Dallas and Ioanis Katakis

Toxins 2025, 17(9), 462; https://doi.org/10.3390/toxins17090462 - 13 Sep 2025

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Abstract

The real-time, cost-effective detection of marine toxins like tetrodotoxin (TTX) remains a significant challenge for the scientific community. Traditional methods, including cell-based assays (CBAs), high-performance liquid chromatography (HPLC), and automated patch clamp (APC), are time-consuming, requiring expensive lab-based equipment and highly trained personnel. [...] Read more.

The real-time, cost-effective detection of marine toxins like tetrodotoxin (TTX) remains a significant challenge for the scientific community. Traditional methods, including cell-based assays (CBAs), high-performance liquid chromatography (HPLC), and automated patch clamp (APC), are time-consuming, requiring expensive lab-based equipment and highly trained personnel. Enzyme-linked immunosorbent assays (ELISAs), lateral flow assays (LFAs), and immunosensors may not be suitable for toxin analogues. Thus, a simplified approach has been developed in this study, which involves the electrophysiological and electrochemical interrogation of N2a cells grown on ITO-coated glass electrodes by measuring extracellular field potentials (EFP) in conjunction with whole-cell patch clamp recordings and electrochemical impedance spectroscopy (EIS) measurements both before and after incubation with TTX. The ITO substrate proved biocompatible and non-toxic for N2a cells. TTX exposure caused 102% inhibition in EFP values at 300 nM, confirmed by sodium current inhibition of 93% at 300 nM and 22% at 1 nM in patch clamp studies (IC₅₀ = 6.7 nM). EIS measurements indicated concentration-dependent impedance changes in the range of 6–300 nM. This research aims to provide a proof-of-concept for integration of electrophysiological and electrochemical approaches to simplify toxin detection systems. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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11 pages, 1642 KB

Open AccessArticle

Sea Anemone-Derived Toxin Avd3i Inhibited Sodium Channel Nav1.4

by Jiaxin Gao, Guohao Liu, Yan Liu, Dezhao Zhang, Qinyi He, Qiong Liao and Canwei Du

Toxins 2025, 17(9), 461; https://doi.org/10.3390/toxins17090461 - 13 Sep 2025

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Abstract

Ion channels regulate ion transport across cell or organelle membranes, playing an important role in various biological processes. Sodium channel Nav1.4 is critical to initiating and propagating action potentials in skeletal muscles, and its dysfunction is associated with a variety of diseases, such [...] Read more.

Ion channels regulate ion transport across cell or organelle membranes, playing an important role in various biological processes. Sodium channel Nav1.4 is critical to initiating and propagating action potentials in skeletal muscles, and its dysfunction is associated with a variety of diseases, such as non-dystrophic myotonias. In this study, U-actitoxin-Avd3i (Avd3i), a Kunitz-type toxin derived from Anemonia viridis, was expressed in prokaryotic systems and was subsequently purified via high-pressure liquid chromatography. Patch clamp recording showed that Avd3i inhibited Nav1.4 in a concentration-dependent manner, with an IC₅₀ of 25.43 μM. However, the toxin exerted no inhibitory activity in Nav1.5/Nav1.7 channels or Kv1.1/Kv1.3/Kv1.4/Kv4.2 potassium channels. Our study found that the sea anemone-derived toxin Avd3i inhibited sodium channel Nav1.4, providing a novel molecule that can act on the channel. Full article

(This article belongs to the Special Issue Peptide Toxins—Discovery, Mechanisms of Action, and Therapeutic Potential)

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19 pages, 2851 KB

Open AccessArticle

Short-Term Effects of Dietary Selenomethionine Supplementation on Hepatic and Renal Transcriptomic Alterations Induced by Ochratoxin a in Broiler Chickens

by Benjamin Kövesi, Szabina Kulcsár, Zsolt Ancsin, Márta Erdélyi, Erika Zándoki, Márk Tóth, Patrik Gömbös, Ágnes Freiler-Nagy, Krisztián Balogh and Miklós Mézes

Toxins 2025, 17(9), 460; https://doi.org/10.3390/toxins17090460 - 12 Sep 2025

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Abstract

Ochratoxin A (OTA), a mycotoxin commonly found in poultry feed, induces oxidative stress and disrupts redox homeostasis in vital organs such as the liver and kidneys. Selenium (Se), an essential trace element, may mitigate OTA-induced toxicity by supporting the antioxidant defense systems. This [...] Read more.

Ochratoxin A (OTA), a mycotoxin commonly found in poultry feed, induces oxidative stress and disrupts redox homeostasis in vital organs such as the liver and kidneys. Selenium (Se), an essential trace element, may mitigate OTA-induced toxicity by supporting the antioxidant defense systems. This study investigated the short-term effects of dietary selenomethionine (SeMet) supplementation on OTA-induced oxidative and transcriptional responses in broiler chickens. Fifty-four 3-week-old birds were fed diets containing 2 mg/kg OTA, a target supplementation of 0.5 mg/kg Se (measured as 0.59 mg/kg as SeMet), or a combination of the two for five days. Liver and kidney samples were collected on Days 1 and 5 for biochemical and gene expression analyses. Exposure to OTA significantly modulated the expression of redox-sensitive transcription factors (NRF2, KEAP1), selenoproteins (GPX3, GPX4, SELK), and detoxification-related genes (AHR, AHRR, CYP1A2). SeMet alone enhanced selenoenzyme expression and antioxidant capacity, while co-exposure partially attenuated OTA-induced oxidative stress, resulting in more pronounced NRF2 activation in the kidneys and CYP1A2 induction in the liver. This is the first study to characterize the transcriptomic responses to OTA exposure in poultry within the first five days, providing novel insight into organ-specific mechanisms and emphasizing the epidemiological relevance of Se supplementation in mitigating the risk of feed contamination. Full article

(This article belongs to the Special Issue Mycotoxins—Biomonitoring and Exposure)

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24 pages, 1195 KB

Open AccessReview

Bacterial Protein Toxins as Anticancer Agents: Clinical Potential of Pseudomonas and Anthrax Toxins

by Richa Misra, Radhika Gupta, Namita Nayyar, Ritvik Baweja, Vishal Sharma, Yogendra Singh and Renu Baweja

Toxins 2025, 17(9), 459; https://doi.org/10.3390/toxins17090459 - 12 Sep 2025

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Abstract

Protein toxins are biologically active polypeptides produced by a variety of organisms, including bacteria, plants, fungi, and animals. These molecules exert potent and specific toxic effects on target cells and are primarily associated with pathogenicity and defense mechanisms of the organisms. In the [...] Read more.

Protein toxins are biologically active polypeptides produced by a variety of organisms, including bacteria, plants, fungi, and animals. These molecules exert potent and specific toxic effects on target cells and are primarily associated with pathogenicity and defense mechanisms of the organisms. In the past few decades, significant progress has been made in understanding their structure, mechanisms of action, and regulation. Among these, bacterial protein toxins have emerged as valuable tools particularly in the development of targeted therapies. A notable example is Botulinum toxin, originally known for its neurotoxic effects, which was approved as a therapeutic agent in 1989 for strabismus treatment, paving way for repurposing bacterial toxins for clinical use. This review provides an overview of the different classes of bacterial toxin-based therapeutics, with a particular focus on Pseudomonas exotoxin A (PE) from Pseudomonas aeruginosa and anthrax toxin from Bacillus anthracis. The modular architecture and potent cytotoxicity of these A-B type toxins have enabled their successful adaptation into targeted cancer therapies. The clinical approval of the PE-based immunotoxin, moxetumomab pasudotox, for the treatment of hairy cell leukemia, underscores the potential of this strategy. This review also discusses current challenges and outlines future directions for the advancement of bacterial toxin-based therapeutics. Full article

(This article belongs to the Section Bacterial Toxins)

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21 pages, 805 KB

Open AccessArticle

Unilateral EMG-Guided Botulinum Toxin for Retrograde Cricopharyngeus Dysfunction: A Prospective Clinical and Neurophysiological Study

by Giuseppe Cosentino, Chiara Zaffina, Clara Zoccola, Mauro Fresia, Sara Merli, Simone Mauramati, Giulia Bertino, Massimiliano Todisco, Shayan Dodge, Sami Barmada, Enrico Alfonsi and Cristina Tassorelli

Toxins 2025, 17(9), 458; https://doi.org/10.3390/toxins17090458 - 12 Sep 2025

Viewed by 388

Abstract

Retrograde cricopharyngeus dysfunction (R-CPD) is a recently recognized condition characterized by the inability to burp, typically accompanied by gurgling noises, bloating, and flatulence. Percutaneous botulinum neurotoxin (BoNT) injection into the cricopharyngeus muscle is a minimally invasive treatment with promising effects, although current evidence [...] Read more.

Retrograde cricopharyngeus dysfunction (R-CPD) is a recently recognized condition characterized by the inability to burp, typically accompanied by gurgling noises, bloating, and flatulence. Percutaneous botulinum neurotoxin (BoNT) injection into the cricopharyngeus muscle is a minimally invasive treatment with promising effects, although current evidence remains limited. In this prospective, open-label study, we evaluated the clinical effects of increasing doses (10 to 30 U) of EMG-guided unilateral BoNT injection in 67 patients with R-CPD. Symptom severity and quality of life were assessed at baseline and at 1 and 4 months post-treatment. The electromyographic (EMG) parameters of the cricopharyngeus were recorded to explore their association with symptom burden and treatment response. At a 1-month follow-up, 55.2% of patients were classified as responders (satisfaction score ≥ 6/10), with a higher rate (64.4%) observed at higher doses, particularly in female patients. Both symptom severity and quality of life improved significantly at 1 month and were sustained at 4 months. Higher cricopharyngeus EMG activity was associated with more severe symptoms and lesser treatment responses. Unilateral EMG-guided BoNT injection is a safe and effective treatment for R-CPD. Further studies should explore the potential role of electromyography in clarifying the pathophysiological aspects of R-CPD and guiding treatment. Full article

(This article belongs to the Special Issue Botulinum Toxins: New Uses in the Treatment of Diseases (2nd Edition))

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14 pages, 2068 KB

Open AccessArticle

Integrative Analysis of Metabolome and Transcriptome Identifies the Role of γ-Glutamylcysteine in Mitigating Deoxynivalenol-Induced Toxicity

by Xiaocheng Bao, Xiaolei Chen, Shuai Chen, Ming-An Sun and Hairui Fan

Toxins 2025, 17(9), 457; https://doi.org/10.3390/toxins17090457 - 12 Sep 2025

Viewed by 299

Abstract

Deoxynivalenol (DON), a prevalent environmental toxin produced by Fusarium fungi, frequently contaminates feed and food products. However, the critical metabolites and regulatory factors involved in DON toxicity remain poorly understood. Building upon our established DON-induced porcine intestinal epithelial cells (IPEC-J2) injury model, this [...] Read more.

Deoxynivalenol (DON), a prevalent environmental toxin produced by Fusarium fungi, frequently contaminates feed and food products. However, the critical metabolites and regulatory factors involved in DON toxicity remain poorly understood. Building upon our established DON-induced porcine intestinal epithelial cells (IPEC-J2) injury model, this study employed liquid chromatography-tandem mass spectrometry (LC-MS/MS) to conduct metabolomic analysis, and integrated analysis with transcriptomic data from DON-exposed IPEC-J2. Results identified 1524 differentially expressed metabolites, and revealed significant enrichment in Glutathione metabolism and Mucin-type O-glycan biosyn-thesis pathways. Notably, γ-glutamylcysteine (γGC), the rate-limiting precursor for glutathione synthesis, showed significant reduction following DON exposure. To explore the biological function of γGC, this study found through exogenous supplementation experiments that γGC pretreatment could significantly alleviate the inhibition of IPEC-J2 cell viability, effectively reduce intracellular ROS accumulation and inhibit DON-induced apoptosis in IPEC-J2 cells. These results indicated that the severe oxidative stress induced by DON is closely related to the blockage of glutathione synthesis caused by the exhaustion of intracellular γGC, and revealed the application potential of γGC as an exogenous supplement in the prevention and treatment of DON exposure. In conclusion, this study offers valuable insights into the metabolic and transcriptional alterations, along with the key metabolites and regulators involved in the cellular response to DON pollution. It also lays a theoretical foundation for more effective prevention and treatment strategies against DON pollution. Full article

(This article belongs to the Special Issue Alleviation of Mycotoxin-Induced Toxicity)

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19 pages, 347 KB

Open AccessArticle

Functional Efficacies of Humate and β-Mannanase Against Aflatoxin B1 and Deoxynivalenol in the Diets for Nursery Pigs

by Yesid R. Garavito-Duarte, Jeonghyeon Son, Alexandra C. Weaver and Sung Woo Kim

Toxins 2025, 17(9), 456; https://doi.org/10.3390/toxins17090456 - 11 Sep 2025

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Abstract

After in vitro mycotoxin binding validation, humate and β-mannanase were tested for mitigating the negative effects of aflatoxin B₁ and deoxynivalenol. Gilts at 8.7 ± 0.5 kg body weight were allotted to four treatments: NC (uncontaminated diet); PC (contaminated diet: 150 µg [...] Read more.

After in vitro mycotoxin binding validation, humate and β-mannanase were tested for mitigating the negative effects of aflatoxin B₁ and deoxynivalenol. Gilts at 8.7 ± 0.5 kg body weight were allotted to four treatments: NC (uncontaminated diet); PC (contaminated diet: 150 µg aflatoxin B₁ and 1100 µg deoxynivalenol per kg feed); HT (PC + humate, 0.5%); and EM (PC + β-mannanase, 800 U/kg diet). Growth performance was recorded for 42 days, and blood and tissue samples were collected for hematological and histopathological evaluations. The PC reduced (p < 0.05) serum tumor necrosis factor-α at day 28 and tended to increase (p = 0.062) immunoglobulin G (IgG), whereas the EM reduced IgG (p < 0.05) at day 42. The PC increased (p < 0.05) mean corpuscular hemoglobin and volume, which were reduced (p < 0.05) by HT or EM at day 42. The PC increased (p < 0.05) bile duct hyperplasia, which was attenuated (p < 0.05) by HT or EM. The PC reduced (p < 0.05) gain- to-feed ratio for the overall period, whereas HT increased (p < 0.05) average daily gain on days 21 to 28. These results suggest that HT and EM may mitigate mycotoxin-induced immune and hepatic damage in pigs through adsorbing mycotoxins. Full article

(This article belongs to the Special Issue Adverse Effects of Co-Contaminated Mycotoxins and Strategies for Their Mitigation)

16 pages, 1087 KB

Open AccessArticle

Development of a Patient-Centered Outcome Tool for Blepharospasm: A Stepwise Modified Delphi Study

by Brian D. Berman, Fares Qeadan, Amanda D. Henderson, Andrew R. Harrison, Giovanni Defazio, Mark Hallett, Gamze Kilic-Berkmen, Laura Wright, Samantha Pentecost, Paul Reyes, Anna Tingin, Joseph Jankovic, Jane Boyd, Charlene Hudgins, Janet Hieshetter, Joel S. Perlmutter, Hyder A. Jinnah and Sarah Pirio Richardson

Toxins 2025, 17(9), 455; https://doi.org/10.3390/toxins17090455 - 10 Sep 2025

Viewed by 559

Abstract

Blepharospasm (BSP) is characterized by excessive orbicularis oculi muscle activity leading to abnormal blinking and involuntary eyelid closure. Botulinum neurotoxin (BoNT) injections are the main treatment for BSP, but they only partially and transiently relieve symptoms, leading to a waxing and waning therapeutic [...] Read more.

Blepharospasm (BSP) is characterized by excessive orbicularis oculi muscle activity leading to abnormal blinking and involuntary eyelid closure. Botulinum neurotoxin (BoNT) injections are the main treatment for BSP, but they only partially and transiently relieve symptoms, leading to a waxing and waning therapeutic response. A patient-centered outcome (PCO) tool that measures BSP symptoms in a simple and efficient way could inform the development of better treatments. Using a stepwise modified Delphi approach, potential PCO items were first identified using the Dystonia Coalition Database with data from over 200 individuals with BSP who had provided responses to existing clinical assessment scales. These items were then analyzed for contribution to overall severity using a Random Forests approach, and redundant items were merged and revised in a series of iterative meetings with a specialist panel along with input from patient advocacy group representatives and focus groups. An online survey was conducted with 330 individuals with BSP to validate and verify the items’ relevance. Finally, the specialist panel provided content validity ratio, which was repeated until it showed good agreement for relevance and clarity of all items. In the end, an easy-to-use PCO tool designed for smartphones and tablets containing 17 items covering three symptom domains (motor, disability, and psychosocial/quality of life) was created. This novel PCO tool for BSP may be used to characterize the cyclical response that an individual patient experiences from BoNT treatments and provide a vital tool for future investigations of longer-acting BoNT preparations or adjunctive therapies. Full article

(This article belongs to the Section Bacterial Toxins)

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27 pages, 4319 KB

Open AccessArticle

In Vitro Screening of the Antifungal and Antimycotoxin Effects of a Stilbenoids-Riche Grapevine Cane Extract on Fusarium graminearum, Aspergillus flavus and Penicillium expansum

by Dorian Aznar, Alexandre Colas de la Noue, Luc P. R. Bidel, Caroline Cayzac, Charlie Poss, Eloïse Ciordia, Andréa Cozette, Angélique Fontana, Fanny Rolet and Caroline Strub

Toxins 2025, 17(9), 454; https://doi.org/10.3390/toxins17090454 - 9 Sep 2025

Viewed by 417

Abstract

Grapevine cane, an abundant viticultural by-product, contains high levels of stilbenoids and therefore holds promise as a natural antifugal and antimycotoxigenic agent. Produced by a microwave-assisted hydro-ethanolic extraction process, the grapevine cane extract (GCE) was tested for its activity against three mycotoxigenic fungi [...] Read more.

Grapevine cane, an abundant viticultural by-product, contains high levels of stilbenoids and therefore holds promise as a natural antifugal and antimycotoxigenic agent. Produced by a microwave-assisted hydro-ethanolic extraction process, the grapevine cane extract (GCE) was tested for its activity against three mycotoxigenic fungi F. graminearum, A. flavus, and P. expansum. Dose-response assays were performed, based on radial growth and inhibition of specific mycotoxin production. For all fungi, growth inhibition IC₅₀ values clustered between 1.0 and 5.0 g/L, while for specific toxin production, IC₅₀ were lower (≈0.5 g/L) except for patulin, which increased in a dose-dependent manner in the presence of the extract. Specific experiments were designed to highlight the effect of the extracts at various stages of the fungal life cycle (e.g., spore germination, early mycelium, and established colonies). F. graminearum spores’ germination was strongly inhibited (5.0 to 15 g/L), while for other fungi, germination was only delayed. Interestingly, antifungal and especially antimycotoxigenic effects were shown to be persistent after exposure. Full article

(This article belongs to the Special Issue Mycotoxins in Food Safety: Challenges and Biocontrol Strategies)

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20 pages, 1235 KB

Open AccessReview

Research Progress on the Detection Methods of Botulinum Neurotoxin

by Shuo Wang, Huajie Zhang, Yanhua Xue, Yingchao Yang and Liyong Yuan

Toxins 2025, 17(9), 453; https://doi.org/10.3390/toxins17090453 - 8 Sep 2025

Viewed by 770

Abstract

Botulinum neurotoxins (BoNTs), produced by the anaerobic spore-forming bacterium Clostridium botulinum, are among the most potent known biological toxins. BoNTs cause lethal botulism via contaminated food, wound infections, or infant intestinal colonization, posing significant threats to public health. Although the mouse bioassay is [...] Read more.

Botulinum neurotoxins (BoNTs), produced by the anaerobic spore-forming bacterium Clostridium botulinum, are among the most potent known biological toxins. BoNTs cause lethal botulism via contaminated food, wound infections, or infant intestinal colonization, posing significant threats to public health. Although the mouse bioassay is still being considered as the gold standard for detecting BoNTs, its drawbacks, including the lengthy experimental duration, high costs, and ethical issues, highlight the urgent need to develop alternative methods to fulfill the detection requirements. In recent years, frequent botulism poisoning incidents haves put forward higher requirements for detection technology. On-site detection is expected to be rapid and immediate, while laboratory detection requires high sensitivity and serotype discrimination capabilities. This review comprehensively introduces current detection approaches, including mouse bioassay, cell-based assays, immunological methods, endopeptidase–mass spectrometry, biosensors, chromatography, and mass spectrometry techniques. Notably, cell-based assays have been used for the potency testing of commercialized botulinum toxin type A and are considered the most promising alternative to the mouse bioassay. Biosensors based on nanomaterials demonstrate advantages in real-time detection due to their rapid response and portability, while endopeptidase–mass spectrometry achieves high sensitivity and effective serotype identification by specifically recognizing toxin-cleaved substrates. Future works shall aim to completely replace MBA, developing a detection system suitable for multiple scenarios such as clinical diagnosis, food safety monitoring, and environmental monitoring. The detection methods should also have matrix compatibility and serotype discrimination capabilities. Full article

(This article belongs to the Section Bacterial Toxins)

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18 pages, 4803 KB

Open AccessArticle

Exploring the Potential of Genista ulicina Phytochemicals as Natural Biocontrol Agents: A Comparative In Vitro and In Silico Analysis

by Roukia Zatout, Ouided Benslama, Fatima Zohra Makhlouf, Alessio Cimmino, Maria Michela Salvatore, Anna Andolfi, Radhia Manel Kolla and Marco Masi

Toxins 2025, 17(9), 452; https://doi.org/10.3390/toxins17090452 - 6 Sep 2025

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Abstract

Development of new sustainable pesticides represents a real challenge for researchers due to environmental issues and public health aspects. In fact, the overuse of chemical pesticides has led to environmental damage, loss of biodiversity, and pesticide-resistant pests. In a framework characterized by the [...] Read more.

Development of new sustainable pesticides represents a real challenge for researchers due to environmental issues and public health aspects. In fact, the overuse of chemical pesticides has led to environmental damage, loss of biodiversity, and pesticide-resistant pests. In a framework characterized by the necessity of new sustainable agricultural practices, this study investigates the plant Genista ulicina as a producer of bioactive compounds for potential application as eco-friendly biopesticides. First, both roots and aerial parts of G. ulicina were extracted and the main compounds in the crude extracts were identified via GC-MS. Subsequently, the crude extracts were submitted to antifungal and phytotoxic assays. In particular, the antifungal effects were evaluated on three common phytopathogenic fungi, Fusarium oxysporum, Alternaria alternata, and Botrytis cinerea, while phytotoxic activity was evaluated on two weed species: Euphorbia peplus L. and Oxalis corniculata L. Further insights were obtained on the herbicidal potential of phytochemical compounds produced by G. ulicina through in silico investigations. In particular, molecular docking analyses were performed against three key enzymes involved in essential plant metabolic pathways: acetohydroxyacid synthase (AHAS), 4-hydroxyphenylpyruvate dioxygenase (HPPD), and protoporphyrinogen oxidase (PPO). Among the compounds identified, linolelaidic acid methyl ester, 1-monolinolein, stearic acid, and palmitic acid derivatives showed promising binding affinities and favorable interaction patterns compared to reference ligands. Selected phytochemicals from G. ulicina show potential as inhibitors of key herbicide targets, suggesting their value as promising leads in the development of sustainable bio-based weed control agents. Full article

(This article belongs to the Section Plant Toxins)

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11 pages, 5432 KB

Open AccessArticle

Galectin-8A Inhibits Cry11Aa Binding to ALP1 and APN 2 Receptors and Toxicity Against Aedes aegypti Larvae

by Xiaohua Hu, Xianhui Huang, Jiannan Liu, Guohui Zhao, Songqing Wu, Xiaoqiang Yu, Lei Xu, Xiong Guan and Lingling Zhang

Toxins 2025, 17(9), 451; https://doi.org/10.3390/toxins17090451 - 6 Sep 2025

Viewed by 422

Abstract

Aedes aegypti, a crucial vector mosquito that transmits many diseases that cause millions of deaths worldwide, can be controlled with Bacillus thuringiensis subsp. israelensis (Bti). The larvicidal activity of Bti against Ae. aegypti is due primarily to Cry4Aa, Cry4Ba, and Cry11Aa, and Cyt1Aa, [...] Read more.

Aedes aegypti, a crucial vector mosquito that transmits many diseases that cause millions of deaths worldwide, can be controlled with Bacillus thuringiensis subsp. israelensis (Bti). The larvicidal activity of Bti against Ae. aegypti is due primarily to Cry4Aa, Cry4Ba, and Cry11Aa, and Cyt1Aa, a protein that synergizes the activity of the Cry proteins. Interestingly, Galectins-6 and Galectins-14, members of a family of β-galactoside-binding proteins that play a role in immune responses insects, have been shown to decrease the activity of Bti toxins. The activity of other Galectins, particularly Galectin-8A, against the Cry proteins is not known. Toward this end, we cloned the gene coding for galactin-8A and expressed the recombinant protein and purified protein. The bioassay results indicated that Galectin-8A can also reduce the toxicity of Cry11Aa, but it was much stronger than Galectin-6. To investigate the interactions among Galectin-8A, Cry11Aa, and toxin receptors, Octet Red System analysis, Western blot, far-Western blot, and ELISA assay were also performed. The Octet Red System result showed that Galectin-8A could also bind to BBMVs of Ae. aegypti, with a lower kDa value than that of Galectin-6, indicating that Galectin-8A had a stronger binding affinity to BBMVs than Galectin-6. Western blot, far-Western blot, and ELISA assay analyses also demonstrated that Galectin-8A bound to Ae. aegypti receptor ALP1 and APN2, consistent with the protein docking simulation results. These findings support the conclusion that Galectin-8A blocks with ALP1 and APN2 more effectively than Galectin-6, which may subsequently reduce the toxicity of Cry11Aa in Ae. aegypti. Full article

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31 pages, 12038 KB

Open AccessArticle

Co-Occurrence of Toxic Bloom-Forming Cyanobacteria Planktothrix, Cyanophage, and Symbiotic Bacteria in Ohio Water Treatment Waste: Implications for Harmful Algal Bloom Management

by Angela Brooke Davis, Morgan Evans, Katelyn McKindles and Jiyoung Lee

Toxins 2025, 17(9), 450; https://doi.org/10.3390/toxins17090450 - 5 Sep 2025

Viewed by 544

Abstract

Cyanobacterial blooms are increasingly becoming more intense and frequent, posing a public health threat globally. Drinking water treatment plants that rely on algal bloom-affected waters may create waste (water treatment residuals, WTRs) that concentrates contaminants. Source waters may contain harmful cyanobacteria, cyanophages (bacteriophages [...] Read more.

Cyanobacterial blooms are increasingly becoming more intense and frequent, posing a public health threat globally. Drinking water treatment plants that rely on algal bloom-affected waters may create waste (water treatment residuals, WTRs) that concentrates contaminants. Source waters may contain harmful cyanobacteria, cyanophages (bacteriophages that infect cyanobacteria), and bacteria. Cyanophages are known to affect bloom formation and growth dynamics, so there is a need to understand viral-host dynamics between phage and bacteria in these ecosystems for managing cyanobacteria. This study isolated and characterized lytic cyanophages from WTRs of a HAB-affected lake in Ohio that infect toxic bloom-forming filamentous cyanobacteria Planktothrix agardhii. Phage infections in the Lake Erie cyanobacteria culture were examined visually and via microscopy and fluorometry. Whole genome sequencing and metagenomic analyses were also conducted. Observed changes in Planktothrix included sheared and shriveled filaments, reduced clumping, and buoyancy changes. Photosynthetic pigmentation was unexpectedly more apparent during phage infection. Metagenomic analyses identified nineteen phages and seven other co-existing bacterial genera. Annotated bacterial genomes contained metabolic pathways that may influence phage infection efficiency. Viral genomes were successfully tied to microbial hosts, and annotations identified important viral infection proteins. This study examines cyanobacterial-phage interactions that may have potential for bioremedial applications. Full article

(This article belongs to the Special Issue Harmful Algal Blooms in Waters: Characterization, Monitoring and Management)

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18 pages, 10242 KB

Open AccessArticle

Toxicity of Volatile Organic Compounds Produced by Pathogens Ewingella americana and Cedecea neteri Associated with Pleurotus pulmonarius

by Zhiyuan Wei, Yifan Wang, Jieheng Qiu, Yulu Nie, Lian Wang and Bin Liu

Toxins 2025, 17(9), 449; https://doi.org/10.3390/toxins17090449 - 5 Sep 2025

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Abstract

Bacterial diseases of Pleurotus pulmonarius, caused by diverse pathogens and associated with a range of symptoms, reduce its commercial value and lead to substantial economic losses. While most research has focused on Pseudomonas tolaasii and its non-volatile toxin tolaasin, little is known [...] Read more.

Bacterial diseases of Pleurotus pulmonarius, caused by diverse pathogens and associated with a range of symptoms, reduce its commercial value and lead to substantial economic losses. While most research has focused on Pseudomonas tolaasii and its non-volatile toxin tolaasin, little is known about other bacterial pathogens and their volatile metabolites. In this study, two bacterial pathogens were isolated from symptomatic P. pulmonarius fruiting bodies in Guangxi, China, and identified as Ewingella americana and Cedecea neteri. Using headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME-GC-MS), we identified 16 volatile organic compounds (VOCs) produced by these two species, seven of which exhibited toxicity-inducing sunken lesions, discoloration, and inhibition of mycelial growth. Symptom severity was quantified by colorimetric analysis. Among the toxic VOCs, 2,4-di-tert-butylphenol was the most potent, inducing sunken lesions and slight discoloration at concentrations as low as 0.5 mg/mL, and causing significant inhibition of mycelial growth at 5 μg/L. The remaining VOCs also caused varying degrees of sunken lesions, yellowing or browning, and suppression of mycelial growth. This study is the first to demonstrate the pathogenic potential of VOCs produced by bacterial pathogens in P. pulmonarius, underscoring their role as important virulence factors and providing a foundation for further investigation into their mechanisms and control strategies. Full article

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18 pages, 2325 KB

Open AccessArticle

Injection of Affibodies by a Self-Organizing Bacterial Syringe to Interfere with Intracellular Signaling

by Thomas Müller, Sophie Gieß, Fanny Maier, Lara Hofacker, Luca Stenger, Larissa Parker, Robert Grosse and Gudula Schmidt

Toxins 2025, 17(9), 448; https://doi.org/10.3390/toxins17090448 - 5 Sep 2025

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Abstract

Photorhabdus luminescens produces a syringe-like toxin complex to inject toxic enzymes into cells. We demonstrated that the recombinant Photorhabdus toxin complex (PTC) can be engineered for translocation of foreign cargo proteins across cellular membranes. We showed that the system is suitable for injection [...] Read more.

Photorhabdus luminescens produces a syringe-like toxin complex to inject toxic enzymes into cells. We demonstrated that the recombinant Photorhabdus toxin complex (PTC) can be engineered for translocation of foreign cargo proteins across cellular membranes. We showed that the system is suitable for injection of trimeric affibodies into mammalian cells in order to influence crucial signaling pathways. As proof of principle, we inhibited Ras-driven tumor cell proliferation by injection of an affibody which interacts with the Ras binding domain of Raf kinase. The system described here could be applicable to target a wide range of signaling molecules for cell biological or therapeutic intervention. Full article

(This article belongs to the Section Bacterial Toxins)

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14 pages, 2477 KB

Open AccessArticle

Potential Linkage Between Zebra Mussel Establishment, Cyanobacterial Community Composition, and Microcystin Levels in United States Lakes

by Feng Zhang, Jayun Kim, Ozeas S. Costa, Jr., Song Liang and Jiyoung Lee

Toxins 2025, 17(9), 447; https://doi.org/10.3390/toxins17090447 - 5 Sep 2025

Viewed by 597

Abstract

Zebra mussel invasion of North American lakes during the last century may play an important role in the occurrence of toxic cyanobacterial blooms. However, empirical evidence quantifying their influence on cyanobacterial community dynamics at broad spatial scales remains limited. Here, we analyzed data [...] Read more.

Zebra mussel invasion of North American lakes during the last century may play an important role in the occurrence of toxic cyanobacterial blooms. However, empirical evidence quantifying their influence on cyanobacterial community dynamics at broad spatial scales remains limited. Here, we analyzed data from the U.S. EPA National Lakes Assessment (>1000 lakes) to examine potential linkages among zebra mussels, cyanobacterial community composition, and cyanotoxin levels. The analysis results showed significant differences in cyanobacterial communities between lakes located in areas with and without established zebra mussel populations. The lakes with established zebra mussels exhibited significantly higher microcystin levels and cyanobacterial abundance, but lower phosphorus concentrations. Structural equation modeling was used to confirm and estimate the effect of zebra mussels on microcystin concentrations via different pathways. The results suggest three potential pathways whereby zebra mussels influence microcystin production: (1) altering phosphorus concentration; (2) increasing cyanobacterial abundance; and (3) shifting cyanobacteria community structure. The total effect of zebra mussel establishment resulted in an overall 1.40-fold net increase in microcystin level, which presumably resulted from three contributing factors: (1) a 1.06-fold increase through an increased cyanobacterial abundance; (2) a 1.53-fold increase through a selective force, resulting in increased cyanobacteria toxicity; and (3) a 0.86-fold decrease in microcystin level through total phosphorus decrease. The study highlights the potential role of zebra mussel invasion in altering cyanobacterial composition and influencing microcystin levels in U.S. lakes. Full article

(This article belongs to the Special Issue Harmful Algal Blooms in Waters: Characterization, Monitoring and Management)

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16 pages, 2449 KB

Open AccessArticle

Comprehensive Insight into Microcystin-Degrading Mechanism of Sphingopyxis sp. m6 Based on Mlr Enzymes

by Qin Ding, Tongtong Liu, Zhuoxiao Li, Rongli Sun, Juan Zhang, Lihong Yin and Yuepu Pu

Toxins 2025, 17(9), 446; https://doi.org/10.3390/toxins17090446 - 5 Sep 2025

Viewed by 611

Abstract

Bacterial degradation is one important Microcystin (MC) removal method in the natural environment. The traditional MC-degrading pathway was proposed based on the functions of individual recombinant Mlr enzymes and the structures of the main MC-degrading products. However, the actual MC-degrading mechanism by Mlr [...] Read more.

Bacterial degradation is one important Microcystin (MC) removal method in the natural environment. The traditional MC-degrading pathway was proposed based on the functions of individual recombinant Mlr enzymes and the structures of the main MC-degrading products. However, the actual MC-degrading mechanism by Mlr enzymes in wild-type bacteria remains unclear. In this study, bioinformatic analysis, heterologous expression, and knockout mutation were performed to elaborate the MC-degrading mechanism by Mlr enzymes in Sphingopyxis sp. m6. The results showed that mlr gene cluster was initially acquired by horizontal gene transfer, followed by vertical inheritance within Alphaproteobacteria. Mlr enzymes exhibit distinct subcellular localizations and possess diverse conserved catalytic domains. The enzymatic cascade MlrA/MlrB/MlrC sequentially cleaves Microcystin-LR (MC-LR) via Adda-Arg, Ala-Leu, and Adda-Glu bonds, generating characteristic intermediates (linearized MC-LR, tetrapeptide, and Adda). Notably, recombinant MlrC demonstrated dual-targeting degrading capability (linearized MC-LR and tetrapeptide), while tetrapeptide specificity in endogenous processing of Sphingopyxis sp. m6. Marker-free knockout mutants of mlr genes were first constructed in MC-degrading bacteria, unveiling that mlrA was indispensable in initial MC cleavage, whereas mlrB/mlrC/mlrD displayed functional compensation through other enzymes with similar functions. This study promotes the mechanistic understanding of MC bacterial degradation and offers a theoretical basis for a bioremediation strategy targeting cyanotoxin pollution. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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17 pages, 835 KB

Open AccessArticle

Application of Graphitized Multi-Walled Carbon Nanotubes Combined with Orbitrap High-Resolution Mass Spectrometry for the Rapid Detection of Ten Toxins in Wild Mushrooms

by Bo Zhang, Yang Liu, Shengnan Li, Ruonan Li, Yunhui Zhang and Hua Zhao

Toxins 2025, 17(9), 445; https://doi.org/10.3390/toxins17090445 - 4 Sep 2025

Viewed by 554

Abstract

Wild mushroom poisoning is an emerging global food safety issue, especially in subtropical regions like southwestern China, where incidents are geographically clustered. Current detection methods are often time-consuming and overlook region-specific toxins. We developed a rapid, sensitive, and accurate method for the simultaneous [...] Read more.

Wild mushroom poisoning is an emerging global food safety issue, especially in subtropical regions like southwestern China, where incidents are geographically clustered. Current detection methods are often time-consuming and overlook region-specific toxins. We developed a rapid, sensitive, and accurate method for the simultaneous detection of ten characteristic mushroom toxins prevalent in Guizhou, China. The method combines graphite multi-walled carbon nanotubes (G-MWCNTs) for sample preparation with Orbitrap high-resolution mass spectrometry (HRMS). Wild mushroom samples were extracted via ultrasonic-assisted methanol–water extraction, purified using G-MWCNTs, and separated on a Hypersil GOLD C18 column (100 mm × 2.1 mm, 1.9 μm). Gradient elution was performed with 0.1% formic acid + 0.01% ammonia and acetonitrile; quantification used the external standard method. The method achieved LODs of 0.005–0.2 mg/kg and LOQs of 0.015–0.6 mg/kg, with RSDs below 18.11% and excellent linearity (R² = 0.9936–0.9989). Among 45 wild mushroom samples, toxin levels ranged from 0.032 to 445.10 mg/kg, with a detection rate of 22.22%, suggesting notable poisoning risk. This method reduces pretreatment time while ensuring high analytical performance, offering a reliable tool for rapid toxin screening and supporting regional surveillance of wild mushroom poisoning. Full article

(This article belongs to the Special Issue Advances in Poisonous Mushrooms and Their Toxins)

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8 pages, 416 KB

Open AccessArticle

Differences in Marine Toxin Poisonings Reported to US Poison Centers After Pandemic Restrictions

by Baylin J. Bennett, Cailee Hill, Hugh B. Roland, Lorraine C. Backer, Amy H. Schnall and Matthew O. Gribble

Toxins 2025, 17(9), 444; https://doi.org/10.3390/toxins17090444 - 4 Sep 2025

Viewed by 480

Abstract

This study investigated whether marine toxin poisonings reported to U.S. Poison Centers changed during the height of the pandemic period (April 2020 to December 2021). The National Poison Data System was queried for single-substance human exposure calls between 1 January 2000 and 31 [...] Read more.

This study investigated whether marine toxin poisonings reported to U.S. Poison Centers changed during the height of the pandemic period (April 2020 to December 2021). The National Poison Data System was queried for single-substance human exposure calls between 1 January 2000 and 31 March 2022 pertaining to ichthyosarcotoxins. Incidence rate ratios for exposure calls were calculated using mixed-effects negative binomial regression. Call counts were aggregated by year and regressed on a binary indicator for occurrence during pandemic restrictions. During the peak pandemic period, exposure calls decreased for several toxins: ciguatera poisoning: 0.57 (0.43, 0.76); clupeotoxic fish poisoning: 0.12 (0.04, 0.39); diarrhetic shellfish poisoning: 0.28 (0.16, 0.49); paralytic shellfish poisoning: 0.23 (0.17, 0.33); scombroid fish poisoning: 0.46 (0.36, 0.57). However, palytoxin poisoning (1.94 (1.32, 2.84)) and tetrodotoxin poisoning (1.73 (1.46, 2.04)) exposure calls appear to have increased. All results were Bonferroni-significant (p ≤ 0.0009). Sensitivity analyses suggest the PLTX increase began prior to pandemic restrictions, whereas the TTX increase appeared to be directly associated with the restrictions. Both men and women reported increases in TTX exposure calls. The TTX increase could be associated with potentially increased participation in outdoor activities, as TTX exposures are linked to amphibia, echinoderms, fish, and mollusks, among other animals. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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Toxins, Volume 17, Issue 9 (September 2025) – 48 articles

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