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Toxins

Journal Description

Toxins

Toxins is an international, peer-reviewed, open access journal related to toxinology and all kinds of toxins (biotoxins) from animals, microbes and plants, and is published monthly online by MDPI. TheFrench Society of Toxinology (SFET)International Society for Mycotoxicology (ISM)Japanese Society of Mycotoxicology (JSMYCO) and European Uremic Toxins (EUTox) Work Group are affiliated withToxins and their members receive a discount on the article processing charges.
  • Open Access— free for readers, witharticle processing charges (APC) paid by authors or their institutions.
  • High Visibility: indexed withinScopus,SCIE (Web of Science),PubMed,MEDLINE,PMC,Embase,CAPlus / SciFinderAGRIS, and other databases.
  • Journal Rank: JCR - Q1 (Toxicology) / CiteScore - Q1 (Toxicology)
  • Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 18.4 days after submission; acceptance to publication is undertaken in 2.8 days (median values for papers published in this journal in the first half of 2025).
  • Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.
  • Sections: published in 6topical sections.
Impact Factor: 4.0 (2024); 5-Year Impact Factor: 4.5 (2024)

Latest Articles

14 pages, 4099 KB  
Article
Structure of Ribosome-Inactivating Protein fromMirabilis jalapa and Its L12-Stalk-Dependent Inhibition ofEscherichia coli Ribosome
byNanami Nishida,Yuki Ninomiya,Toru Yoshida,Takehito Tanzawa,Yasushi Maki,Hideji Yoshida,Hideaki Tsuge andNoriyuki Habuka
Toxins2025,17(12), 575;https://doi.org/10.3390/toxins17120575 - 28 Nov 2025
Abstract
Mirabilis antiviral protein (MAP) is the type I ribosome-inactivating protein (RIP), which consists of an RNAN-glycosylase domain with no carbohydrate-binding domain. Unlike many RIPs, such as ricin or trichosanthin, which inactivate eukaryotic ribosomes, MAP also inactivates theE. coli ribosome by [...] Read more.
Mirabilis antiviral protein (MAP) is the type I ribosome-inactivating protein (RIP), which consists of an RNAN-glycosylase domain with no carbohydrate-binding domain. Unlike many RIPs, such as ricin or trichosanthin, which inactivate eukaryotic ribosomes, MAP also inactivates theE. coli ribosome by cleaving theN-glycosidic bond at A2660 of 23S ribosomal RNA. The structure of the wild-type MAP has not been revealed yet. Here, we expressed, purified, and crystallized the plural recombinant MAPs, including both E168Q and R171Q mutations (MAP-EQRQ) inE. coli, and determined the crystal structure of MAP-EQRQ at 2.1 Å resolution. According to the predicted structure with RNA (sarcin-ricin loop) and the mutant protein’s activities using quantitative RT-PCR, we showed that residue R171 at the active site of MAP is a key residue to form the stable complex with target adenine. Furthermore, we showed that MAP bound the C-terminal domains of eukaryotic P2-stalk as well asE. coli L12-stalk.Full article
(This article belongs to the SectionPlant Toxins)
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19 pages, 1624 KB  
Article
Mycotoxin-Induced Oxidative Stress and Its Impact on Human Folliculogenesis: Examining the Link to Reproductive Health
byZsuzsanna Szőke,Eszter Ruff,Patrik Plank,Zsófia Molnár,Lili Hruby,Apolka Szentirmay,Márkó Unicsovics,Bernadett Csókay,Katalin Varga,Tímea Buzder,Miklós Sipos,Katalin Sára-Popovics,Dóra Holéci,Katalin Posta andLevente Sára
Toxins2025,17(12), 574;https://doi.org/10.3390/toxins17120574 - 28 Nov 2025
Abstract
Climate change has contributed to increased mycotoxin contamination in food systems, posing a growing threat to human health, including reproductive health. Our study aimed to investigate how mycotoxins entering the follicular fluid affect oxidative stress processes. We analyzed 88 follicular fluid samples from [...] Read more.
Climate change has contributed to increased mycotoxin contamination in food systems, posing a growing threat to human health, including reproductive health. Our study aimed to investigate how mycotoxins entering the follicular fluid affect oxidative stress processes. We analyzed 88 follicular fluid samples from infertile patients for common mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), its main metabolite alpha-zearalenol (aZOL), and aflatoxin M1 (AfM1), and examined their relationship with oxidative stress markers (MDA, SOD, GPx, CAT, and TAOC) and hormones (cortisol, estradiol, and anti-Müllerian hormone). Higher mycotoxin levels were associated with increased oxidative stress, particularly elevated MDA levels, and disrupted antioxidant enzyme activity. Notably, DON showed a positive correlation with SOD and estradiol levels, indicating a compensatory antioxidant response, while AfM1 served as a negative predictor. The metabolite aZOL was strongly linked to cortisol, with effects influenced by estradiol levels, implying endocrine-disrupting activity. Importantly, the interaction between DON and AMH appeared to impact dominant follicle development, suggesting a potential mechanism by which environmental toxins impair fertility without directly reducing oocyte or embryo counts. These results highlight the complex, dose-dependent effects of mycotoxins on oxidative and hormonal balances within the follicular environment, with implications for oocyte quality and reproductive success. Better understanding these mechanisms could help develop early diagnostic markers and targeted interventions to improve fertility outcomes in women exposed to changing environmental conditions.Full article
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12 pages, 254 KB  
Communication
Artificial Intelligence in Managing Spasticity with Botulinum Toxin Type A—Insights from an Exploratory Pilot Investigation: The AIMS Study
byMirko Filippetti,Rita Di Censo,Lyria Arcari,Maria Concetta Schiavariello,Marco Battaglia,Salvatore Facciorusso,Stefania Spina,Laura Antonucci,Andrea Santamato,Alessio Baricich,Nicola Smania andAlessandro Picelli
Toxins2025,17(12), 573;https://doi.org/10.3390/toxins17120573 - 27 Nov 2025
Abstract
This study aimed to explore the potential role of artificial intelligence in optimizing botulinum toxin type A treatment for spasticity and to evaluate its alignment with expert clinical decisions. A comparative analysis was conducted using thirty hypothetical clinical cases involving individuals with spasticity [...] Read more.
This study aimed to explore the potential role of artificial intelligence in optimizing botulinum toxin type A treatment for spasticity and to evaluate its alignment with expert clinical decisions. A comparative analysis was conducted using thirty hypothetical clinical cases involving individuals with spasticity resulting from various neurological conditions. Five rehabilitation physicians, each with more than five years of experience, participated in the study. An artificial intelligence model trained on scientific literature and clinical guidelines generated treatment recommendations, including target muscles and dosages, which were compared with those proposed independently by the physicians. The primary outcome was the level of agreement in muscle selection and dosage. The model demonstrated consistency and adherence to guidelines but showed limited adaptability in complex presentations, such as an adducted thigh and equinovarus foot. It generally recommended lower dosages and differed significantly from physicians in both muscle selection and treatment strategies. Artificial intelligence shows promise as a clinical support tool in spasticity management, offering standardized and reproducible recommendations. However, its limited capacity to interpret clinical subtleties currently restricts its practical application. Future models should integrate multimodal clinical data and real-time clinician feedback to better emulate expert decision-making processes.Full article
14 pages, 1561 KB  
Systematic Review
Impact of Protein Concentrate Production on Mycotoxin Mitigation: A Systematic Review
byCaroline Senna,Marianna Cruz,Larine Kupski andEliana Badiale-Furlong
Toxins2025,17(12), 572;https://doi.org/10.3390/toxins17120572 - 26 Nov 2025
Abstract
The plant-based protein industry has explored new material sources, such as agro-industrial by-products and extraction techniques based on chemical properties assisted by ultrasound, high pressure and other tools to improve the yield and functionality of protein concentrates. However, promising by-products from vegetable processing [...] Read more.
The plant-based protein industry has explored new material sources, such as agro-industrial by-products and extraction techniques based on chemical properties assisted by ultrasound, high pressure and other tools to improve the yield and functionality of protein concentrates. However, promising by-products from vegetable processing are susceptible to incidental and natural contaminants, mainly mycotoxins. Adopting sustainable strategies and understanding how they affect mycotoxin fate during processing remains a challenge to ensure food security. In this study, a systematic literature review and bibliometric analysis were conducted to identify reliable pre-treatments and treatments for producing protein concentrates and evaluate the efficiency of technologies to mitigate mycotoxin bioaccessibility. Searching for research in Scopus, Web of Science and ScienceDirect (2010–2024) identified 3688 scientific articles on techniques to improve the yield and functionality of recovered proteins, but only three studies addressed mycotoxin fate. Aflatoxin, the most prevalent mycotoxin in raw materials, was the only one considered, highlighting that chemical and enzymatic treatments may help mitigate mycotoxicological risks in protein concentrates. Results indicate a gap in plant-based food security regarding mycotoxin contamination, which must be addressed through mitigation strategies aligned with efficient processes to ensure sustainable and safe plant protein-based foods.Full article
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18 pages, 5407 KB  
Article
Differential Effects of Marimastat and Prinomastat on the Metalloprotease Activity of Various Snake Venoms
byMahtab Khatibi,José R. Almeida,Soheil Gilabadi,David Ramírez,Paulina Valenzuela-Hormazábal,Ketan Patel andSakthivel Vaiyapuri
Toxins2025,17(12), 571;https://doi.org/10.3390/toxins17120571 - 26 Nov 2025
Abstract
Snakebite envenoming is a neglected tropical disease, responsible for approximately 140,000 deaths globally each year. Vipers and elapid snakes represent the most significant snake families in medical contexts, exhibiting a variety of venom components and clinical effects in bite victims. Metalloproteases, a primary [...] Read more.
Snakebite envenoming is a neglected tropical disease, responsible for approximately 140,000 deaths globally each year. Vipers and elapid snakes represent the most significant snake families in medical contexts, exhibiting a variety of venom components and clinical effects in bite victims. Metalloproteases, a primary component of venoms, are mainly accountable for haemotoxic and myotoxic effects. Although predominantly found in viper venoms, these enzymes are also present in varying levels in elapid snake venoms. Marimastat and prinomastat are matrix metalloprotease inhibitors initially developed as cancer therapies. Recently, extensive research has focused on these inhibitors to neutralise venom metalloproteases. However, their effects on different viper and elapid snake venoms remain unclear. Here, we report the sensitivity of seven elapid venoms (specifically, cobras) and 12 viper venoms to marimastat and prinomastat, utilising selective in vitro experiments and molecular docking analyses performed using representative metalloprotease (VAP2, a viper metalloprotease from the venom ofCrotalus atrox and an elapid metalloprotease from the venom ofNaja atra) structures. Both compounds inhibited the metalloprotease, fibrinogenolytic, and caseinolytic activities of most viper venoms. While prinomastat displayed prominent inhibitory effects on cobra venoms in these assays, marimastat demonstrated limited inhibitory effects on these venoms. These findings illustrate the role of matrix metalloprotease inhibitors in modulating metalloprotease activities across a range of viper and cobra venoms. Collectively, this study establishes the differential effects of marimastat and prinomastat on various levels of metalloproteases present in viper and elapid venoms. This will enhance understanding of the abundance of metalloproteases in snake venoms and their sensitivity to different matrix metalloprotease inhibitors.Full article
(This article belongs to the Special IssueBiochemistry, Pathology and Applications of Venoms)
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18 pages, 2187 KB  
Article
AcuteAmanita muscaria Toxicity: A Literature Review and Two Case Reports in Elderly Spouses Following Home Preparation
byStanila Stoeva-Grigorova,Ivanesa Yarabanova,Maya Radeva-Ilieva,Diana Ivanova,Snezha Zlateva andPetko Marinov
Toxins2025,17(12), 570;https://doi.org/10.3390/toxins17120570 - 25 Nov 2025
Abstract
Amanita muscaria (L.) Lam., commonly known as fly agaric, remains an uncommon yet clinically important cause of acute mushroom intoxication. Although typically associated with mild to moderate neuropsychiatric disturbances, the mushroom’s toxic profile is highly variable and continues to attract scientific, toxicological, [...] Read more.
Amanita muscaria (L.) Lam., commonly known as fly agaric, remains an uncommon yet clinically important cause of acute mushroom intoxication. Although typically associated with mild to moderate neuropsychiatric disturbances, the mushroom’s toxic profile is highly variable and continues to attract scientific, toxicological, and public health interest. This work provides an integrative review of the biochemical composition, toxicodynamics, and clinical manifestations associated withA. muscaria exposure, with particular emphasis on the pharmacological actions of its principal constituents, ibotenic acid and muscimol. The review is complemented by two contemporaneous cases of severe intoxication in elderly individuals, illustrating the real-world clinical expression of the toxidrome and the challenges in diagnosis and management. Both cases presented with rapid-onset gastrointestinal symptoms, profound central nervous system depression, and cholinergic features, requiring intensive supportive therapy, atropine infusion, and continuous monitoring. Full recovery was achieved in both patients. These clinical observations contextualize the broader toxicological framework discussed in the review and underscore the need for increased clinical vigilance, improved public education, and strengthened diagnostic and therapeutic preparedness regarding psychoactive wild mushroom exposures.Full article
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13 pages, 1185 KB  
Systematic Review
Botulinum Toxin Combined with Robot-Assisted Therapy for Post-Stroke Spasticity: A Systematic Review
bySalvatore Facciorusso,Stefania Spina,Mirko Filippetti,Rajiv Reebye,Gerard E. Francisco andAndrea Santamato
Toxins2025,17(12), 569;https://doi.org/10.3390/toxins17120569 - 25 Nov 2025
Abstract
(1) Background: Post-stroke spasticity limits motor recovery and independence. Combining botulinum toxin type-A (BoNT-A) injection with intensive, task-specific robot-assisted therapy (RAT) might enhance neuroplasticity and functional gains, but its additive effect and optimal timing are uncertain. (2) Methods: We systematically searched major medical [...] Read more.
(1) Background: Post-stroke spasticity limits motor recovery and independence. Combining botulinum toxin type-A (BoNT-A) injection with intensive, task-specific robot-assisted therapy (RAT) might enhance neuroplasticity and functional gains, but its additive effect and optimal timing are uncertain. (2) Methods: We systematically searched major medical databases and trial registries up to April 2025 for randomized controlled trials in adults with post-stroke spasticity comparing botulinum toxin type-A injection plus RAT with toxin injection plus conventional therapy, or RAT alone with RAT combined with toxin injection. Risk of bias was assessed using the RoB 2 tool, and findings were synthesized narratively. (3) Results: Seven trials (n = 229) were included. Across all studies, toxin treatment reduced spasticity within groups, whereas additional spasticity reduction with RAT versus conventional rehabilitation was inconsistent. In contrast, several lower-limb trials reported greater improvements in walking capacity and balance when RAT was added, while upper-limb trials showed comparable motor recovery across treatment arms with occasional advantages in strength and movement quality. A pilot four-arm study suggested that starting RAT around four weeks after injection may maximize upper-limb motor gains. (4) Conclusions: The combination of BoNT-A with RAT appears safe and is particularly promising for gait rehabilitation, but further research is needed to define optimal timing and protocols.Full article
(This article belongs to the SectionBacterial Toxins)
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16 pages, 1365 KB  
Article
Dose-Dependent but Non-Interactive Effects of Ochratoxin A and Selenomethionine on Hepatic Lipid Metabolism and Oxidative Stress in Broiler Chickens
bySzabina Kulcsár,Krisztián Balogh,Erika Zándoki,Edward Agyarko,Omeralfaroug Ali,Benjámin Kövesi,Ágnes Freiler-Nagy,András Szabó andMiklós Mézes
Toxins2025,17(12), 568;https://doi.org/10.3390/toxins17120568 - 25 Nov 2025
Abstract
This study examined the effects of ochratoxin A (OTA) exposure and graded dietary selenium (Se) supplementation on fatty acid (FA) composition and oxidative stress markers in the liver of broiler chickens. OTA is known to generate oxidative stress, promote lipid peroxidation, and affect [...] Read more.
This study examined the effects of ochratoxin A (OTA) exposure and graded dietary selenium (Se) supplementation on fatty acid (FA) composition and oxidative stress markers in the liver of broiler chickens. OTA is known to generate oxidative stress, promote lipid peroxidation, and affect the antioxidant system. Se, an essential trace element with antioxidant properties, may help counteract OTA-induced toxicity. In this short-term (5-day) in vivo feeding experiment, 21-day-old broiler chickens were divided into six groups, each with six birds: Control (diet free from Se), 0.3 mg/kg Se, 0.5 mg/kg Se, 2 mg/kg OTA, 2 mg/kg OTA + 0.3 mg/kg Se, 2 mg/kg OTA + 0.5 mg/kg Se. Our findings show that supplementing 0.3 mg/kg (p < 0.01) or 0.5 mg/kg Se (p < 0.001) in OTA-exposed birds significantly reduced the early oxidative stress markers (conjugated dienes and trienes) and significantly increased (0.3 mg/kgp < 0.01; 0.5 mg/kgp < 0.001) glutathione levels, indicating enhanced glutathione-dependent antioxidant protection. The treatments also significantly altered the ratio of monounsaturated and n6/n3 polyunsaturated FAs. OTA with 0.3 mg/kg Se supplementation significantly (p ˂ 0.05) reduced total unsaturation and FA average chain length. At a dose of 0.3 mg/kg, the interaction of Se and OTA altered the PUFA composition, while 0.5 mg/kg Se supplementation enhanced antioxidant defense and reduced lipid peroxidation. These results highlight the dual but separate role of Se, where inadequate doses may enhance OTA toxicity, while optimal supplementation may have a protective effect on hepatic lipid homeostasis. These findings can be used in the future progress of the mitigation strategy against OTA exposure in poultry nutrition.Full article
(This article belongs to the Special IssueMycotoxins—Biomonitoring and Exposure)
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17 pages, 2236 KB  
Review
Updates on the Proposed Botulinum Toxin A Mechanisms of Action in Orofacial Pain: A Review of Animal Studies
byJaime Fabillar Jr,Yumiko Yamamoto,Kazuyuki Koike,Daisuke Ikutame andYoshizo Matsuka
Toxins2025,17(12), 567;https://doi.org/10.3390/toxins17120567 - 23 Nov 2025
Abstract
Experimental animal models of orofacial pain have been instrumental in elucidating biological pathways underlying the antinociceptive effect of botulinum neurotoxin type A (BoNT/A). Although several mechanisms have been proposed to explain how BoNT/A relieves pain, the precise modes of action, particularly in the [...] Read more.
Experimental animal models of orofacial pain have been instrumental in elucidating biological pathways underlying the antinociceptive effect of botulinum neurotoxin type A (BoNT/A). Although several mechanisms have been proposed to explain how BoNT/A relieves pain, the precise modes of action, particularly in the oral and maxillofacial areas, remain elusive. The purpose of this review was to synthesize and assess the latest proposed mechanisms of action through which BoNT/A attenuates orofacial pain in established animal models. A comprehensive search was conducted using the terms “botulinum neurotoxin,” “mechanism,” and “orofacial pain” or “trigeminal neuralgia.” Only animal studies involving the establishment of an orofacial pain model were selected. Additional relevant studies were identified through manual screening of cited references. Over the past five years, several animal pain models have been established to decipher the mechanisms underlying the BoNT/A-mediated antinociception in orofacial pain. The proposed mechanisms include retrograde transport, neuronal excitability regulation, neuropeptide inhibition, inflammatory modulation, and opioid system stimulation in both the peripheral and central nervous systems. Despite the insubstantial number of investigations and findings, BoNT/A exhibits multidimensional modulation of nociceptive responses and, therefore, remains a promising therapeutic agent for managing orofacial pain conditions, with animal studies consistently providing insights into the mechanism of its antinociceptive action.Full article
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17 pages, 548 KB  
Article
Fumonisin Intake from Consumption of Wheat- and Corn-Based Products in Hungary
byTamás Schieszl,Ákos Jozwiak,Miklós Süth,Imre Nemes,Melinda Kovács andAndrea Zentai
Toxins2025,17(12), 566;https://doi.org/10.3390/toxins17120566 - 22 Nov 2025
Abstract
Fumonisins are mycotoxins commonly found on corn and other grains, and have been linked to several health concerns. The aim of this study is to estimate the exposure of the Hungarian population to fumonisins from regular food consumption. Fumonisin B1 and B2 concentrations [...] Read more.
Fumonisins are mycotoxins commonly found on corn and other grains, and have been linked to several health concerns. The aim of this study is to estimate the exposure of the Hungarian population to fumonisins from regular food consumption. Fumonisin B1 and B2 concentrations were determined in commercially available corn-, wheat- and rice-based foods. Daily intakes on an individual level were calculated deterministically based on recent individual food consumption data distributions, and the average contamination of the concerned food categories. The most significant sources of fumonisins were corn flour, cornmeal, cornflakes, other corn-based products, and wheat-based products (average total fumonisin contents in the middle bound scenario were 0.115, 0.074, 0.052, 0.091 and 0.077 mg/kg, respectively). In cases where the concentration values measured below the analytical determination limits, these were substituted by half of the corresponding limits (middle bound scenario). Mean and high (95th percentile) exposures to fumonisins B1 + B2 were 0.101 and 0.258 µg/bw kg/day for adults, and 0.313 and 0.744 µg/bw kg/day for children, respectively. Our results show that about 1.5% of children’s exposure could exceed the tolerable daily intake limit (TDI) of fumonisins, established by EFSA in 2018, meaning that potential health risks cannot be ruled out for a low proportion of consumers. We found that besides corn-based products, wheat-based food products could also contribute to total intake, due to their high consumption levels. Constant monitoring of fumonisin levels in corn- and wheat-based products is recommended to safeguard consumer health.Full article
(This article belongs to the SectionMycotoxins)
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13 pages, 5739 KB  
Article
Bacterial Microbiota in Soil Amended with Deoxynivalenol-Contaminated Wheat
byEmmanuel W. Bumunang,Kim Stanford,Yuxi Wang,Benjamin Ellert,Matthew Waldner andTrevor W. Alexander
Toxins2025,17(12), 565;https://doi.org/10.3390/toxins17120565 - 22 Nov 2025
Abstract
Feed contaminated with the mycotoxin deoxynivalenol (DON) can negatively impact livestock health and performance. Bacteria capable of degrading DON present a method of mitigating its harmful effects. This study aimed to identify microbial consortia from soil samples that could degrade DON. Soil from [...] Read more.
Feed contaminated with the mycotoxin deoxynivalenol (DON) can negatively impact livestock health and performance. Bacteria capable of degrading DON present a method of mitigating its harmful effects. This study aimed to identify microbial consortia from soil samples that could degrade DON. Soil from central (Lacombe, LA) and southern (Lethbridge, LE) Alberta were used as microbial inoculant. The soils were mixed with DON-contaminated wheat (18 ppm/kg) on day 0, and each soil type was divided into triplicate pots (180 g) and placed in a controlled environment for 32 d. Control pots of each soil type were included, which contained no DON-contaminated wheat. On days 0, 7, 14, and 32, 1 g subsamples were collected from pots, serially diluted in a limited medium containing DON (10 µg/mL) as the only carbon source, and incubated for 2 weeks (30 °C). DNA was extracted from the pots across time, as well as the subsample consortia grown in DON-amended medium, and was analyzed for bacterial changes after 16S rRNA gene sequencing. The relative abundance of bacterial genera in soil samples after enrichment with DON-contaminated wheat increased across time compared to the baseline day 0 time point. DON-degrading activity (26%) was only detected in LA soil suspension on day 7, and was highest after 14 days of incubation. The most abundant bacteria in the LA DON-degrading consortia belonged to thePseudomonas (8.8%),Delftia (7.4%),Acinetobacter (6.4%),Comamonas (5.7%),Stenotrophomonas (5.5%),Shinella (5.5%),Ensifer (5.1%),Agrobacterium (5.0%),Achromobacter (4.7%), andRhizobium (3.7%) genera.Pseudomonas aeruginosa (n = 9) andSerratia liquefaciens (n = 3) strains isolated from the LA consortia did not degrade DON. Overall, this study shows that the soil contained bacteria capable of degrading DON; however, variation existed depending on the soil’s source.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
byYubo Song,Hao Wang,Junjie Wen,Jiale Xu,Siyu Zhu,Fuli Wang andYongqian Zhang
Toxins2025,17(11), 564;https://doi.org/10.3390/toxins17110564 - 18 Nov 2025
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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17 pages, 6436 KB  
Article
Cytotoxic Effects Induced by Combined Exposure of the Patulin, Ochratoxin A, and Acetamiprid to HK-2 and SK-N-SH Cell Lines
byZuoyin Zhu,Hanke Zhao,Xianli Yang,Dongxia Nie,Junhua Yang andZheng Han
Toxins2025,17(11), 563;https://doi.org/10.3390/toxins17110563 - 18 Nov 2025
Abstract
Patulin (PAT), ochratoxin A (OTA), and acetamiprid (ACM) are common food contaminants that frequently co-occur in agricultural products, raising concerns over their cumulative health risks. This study is the first to systematically assess the combined cytotoxic effects of PAT, OTA, and ACM using [...] Read more.
Patulin (PAT), ochratoxin A (OTA), and acetamiprid (ACM) are common food contaminants that frequently co-occur in agricultural products, raising concerns over their cumulative health risks. This study is the first to systematically assess the combined cytotoxic effects of PAT, OTA, and ACM using combination index (CI) and dose reduction index (DRI) models in HK-2 and SK-N-SH cells. All three compounds exhibited dose-dependent toxicity, with potency ranked as PAT > OTA > ACM. In HK-2 cells, PAT+OTA and OTA+ACM showed primarily antagonistic interactions, with synergism observed at low doses. PAT+ACM displayed exposure time-dependent additive effects, while the ternary mixture was mostly antagonistic, with OTA being the dominant contributor. In SK-N-SH cells, most combinations were antagonistic; however, OTA+ACM showed dose-dependent shifts, and the triple mixture transitioned from antagonism to synergism at higher concentrations. OTA and ACM were identified as the main toxicity drivers in all combinations. These findings highlight dose- and cell-specific interactions and underscore the importance of cumulative risk assessment of co-occurring mycotoxins and pesticides in food safety regulation.Full article
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18 pages, 5231 KB  
Article
Trace Aflatoxins Extraction in Pistachio, Maize and Rice Based on β-Cyclodextrin-Doped Cu-Carboxylated Graphene Oxide Nanocomposite
byAmr A. Yakout,Wael H. Alshitari,Hassan M. Albishri,Faten M. Ali Zainy andAdel M. Alshutairi
Toxins2025,17(11), 562;https://doi.org/10.3390/toxins17110562 - 17 Nov 2025
Abstract
Aflatoxins remain among the most challenging food contaminants to monitor due to their structural diversity, low abundance, and the chemical complexity of cereal- and nut-based matrices. In this study, a multifunctional Cu/β-cyclodextrin@carboxylated graphene oxide (Cu/β-CD@CGO) nanocomposite was synthesized through a green, two-step procedure [...] Read more.
Aflatoxins remain among the most challenging food contaminants to monitor due to their structural diversity, low abundance, and the chemical complexity of cereal- and nut-based matrices. In this study, a multifunctional Cu/β-cyclodextrin@carboxylated graphene oxide (Cu/β-CD@CGO) nanocomposite was synthesized through a green, two-step procedure and employed as a high-affinity nanosorbent for trace extraction of AFB1, AFB2, AFG1, and AFG2. The architecture integrates three complementary components: β-cyclodextrin for inclusion-driven molecular recognition, copper nanoparticles that establish coordination interactions with lactone-bearing aflatoxins, and CGO nanosheets that supply extensive π-rich surfaces and abundant carboxyl functionalities. Comprehensive characterization (FTIR, Raman, XPS, SEM, EDX-mapping, and HRTEM) confirmed the formation of a uniform, porous hybrid network. Under optimized d-SPE conditions, the nanocomposite enabled quantitative recovery (92.0–108.5%) of aflatoxins from pistachio, maize, and rice extracts while achieving sub-ng kg−1 detection limits and excellent reproducibility. The results demonstrate that the Cu/β-CD@CGO platform provides a robust, selective, and sustainable alternative to conventional immunoaffinity or polymeric sorbents, offering strong potential for routine surveillance of aflatoxins in complex food systems.Full article
(This article belongs to the SectionMycotoxins)
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18 pages, 2788 KB  
Article
Venom Peptides Across Asian and American Tarantulas Utilize Dual Pharmacology to Target Activation and Fast Inactivation of Voltage-Gated Sodium Channels
byAmatulla S. Nashikwala,Charan Kotapati,David A. Eagles,Richard J. Lewis andFernanda C. Cardoso
Toxins2025,17(11), 561;https://doi.org/10.3390/toxins17110561 - 14 Nov 2025
Abstract
Spider-derived venoms are a rich source of cystine knot peptides with immense therapeutic potential. Many of these peptides exert unique biological activities through the modulation of ion channels, including of human voltage-gated sodium (NaV1.1–NaV1.9) channels. NaV channel subtypes [...] Read more.
Spider-derived venoms are a rich source of cystine knot peptides with immense therapeutic potential. Many of these peptides exert unique biological activities through the modulation of ion channels, including of human voltage-gated sodium (NaV1.1–NaV1.9) channels. NaV channel subtypes have diverse functions determined by their tissue and cellular distribution and biophysical properties, and are pathophysiology mediators in various diseases. Therefore, NaVs are central in studies of human biology. This work investigated the pharmacological properties of venom of the Thai theraphosidOrnithoctonus aureotibialis on NaV channels. We discovered a predominant venom peptide named Oa1a and assessed its pharmacological properties across human NaV channel subtypes. Synthetic forms of the peptide Oa1a showed preferential inhibition of NaV1.1 and NaV1.7, while recombinant Oa1a displayed a preference for inhibiting NaV1.2, NaV1.6, and NaV1.7. Interestingly, all versions of Oa1a peptides exerted dual pharmacological effect by reducing the peak current and slowing fast inactivation of NaV1.3, consistent with Oa1a having more than one binding site on NaV channels. Such complex pharmacology was previously observed for a venom peptide in a Central American and Costa Rican tarantula, suggesting a conserved mechanism of action amongst these geographically distinct species. However, Oa1a lacked activity in the T-type channels observed in the tarantula peptide from Central America. Structure–function relationships investigated using molecular modelling showed that the dual pharmacology is driven by a conserved mechanism utilizing a mix of aromatic and charged residues, while the T-type activity appears to require additional charged residues in loop 2 and fewer positive charges in loop 4. Future structure–activity relationship studies of Oa1a will guide the development of pharmacological tools as well as next-generation drugs to treat NaV channel dysfunction associated with neurological disorders.Full article
(This article belongs to the SectionAnimal Venoms)
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20 pages, 3294 KB  
Article
Brevetoxin Dynamics and Bioavailability from Floc Following PAC-Modified Clay Treatment ofKarenia brevis Blooms
byNicholas R. Ohnikian,Christopher D. Sibley,R. Ben Freiberger,Kristen N. Buck,Alyssa Myers,Samantha Harlow,Donald M. Anderson,Richard Pierce andJennifer H. Toyoda
Toxins2025,17(11), 560;https://doi.org/10.3390/toxins17110560 - 13 Nov 2025
Abstract
Harmful algal blooms (HABs) caused by the dinoflagellateKarenia brevis present serious ecological and public health concerns due to the production of brevetoxins (BTX). Clay flocculation and sedimentation of cells, particularly with polyaluminum chloride (PAC)-modified clays, is a promising HAB mitigation approach. This [...] Read more.
Harmful algal blooms (HABs) caused by the dinoflagellateKarenia brevis present serious ecological and public health concerns due to the production of brevetoxins (BTX). Clay flocculation and sedimentation of cells, particularly with polyaluminum chloride (PAC)-modified clays, is a promising HAB mitigation approach. This study evaluated the efficacy of Modified Clay-II (MCII), a PAC-modified kaolinite clay, in reducingK. brevis cell abundance in mesocosm experiments and examined the bioavailability of BTX potentially released from settled floc back into the water column and sediment over the first 72 h after treatment. Additionally, we quantified trace metals in benthic clams (Mercenaria mercenaria) exposed to the floc post-treatment to assess metal accumulation and potential toxicological effects from MCII application. MCII treatment (0.2 g/L) resulted in a 91% reduction inK. brevis cell density and a 50% decrease in waterborne brevetoxins after 5 h. Brevetoxins accumulated in sediment post-flocculation, with BTX-B5 emerging as the dominant congener. Clams exposed to MCII-treated floc showed comparable tissue BTX levels to controls and significantly elevated aluminum concentrations, though without mortality. The aluminum accumulations in this study do not raise concerns for the health of the clams or the humans who eat them, given other dietary exposures. These findings support the potential of MCII for HAB mitigation while underscoring the need for further evaluation of exposure risks to all benthic species.Full article
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23 pages, 11689 KB  
Article
Visual Tracking and Organ Targeting ofNaja atra andDeinagkistrodon acutus Venoms in Mice
byShaocong Hu,Manqi Xiao,Ningjing Jiang,Ziyan Zhang,Qiuju Jia,Yi Zhou,Xin Liu andMing Liao
Toxins2025,17(11), 559;https://doi.org/10.3390/toxins17110559 - 13 Nov 2025
Abstract
In China, bites caused by theNaja atra andDeinagkistrodona acutus are the most common types of snakebites. While the functional characteristics of the two venom components have been well documented, their in vivo metabolic pathways, target organ distribution patterns, and dynamic pharmacokinetic [...] Read more.
In China, bites caused by theNaja atra andDeinagkistrodona acutus are the most common types of snakebites. While the functional characteristics of the two venom components have been well documented, their in vivo metabolic pathways, target organ distribution patterns, and dynamic pharmacokinetic profiles remain less explored. This study established a murine envenoming model through CY7-SE labeling ofNaja atra andDeinagkistrodon acutus venoms. The real-time in vivo absorption and biodistribution of venoms were dynamically monitored via fluorescence imaging, with subsequent proteomic profiling to characterize organ-specific toxin targeting patterns. Gel filtration chromatography and HPLC analyses validated labeling efficiency at ratios of 0.1 mg CY7-SE per 1 mgNaja atra venom and 0.075 mg CY7-SE per 1 mgDeinagkistrodon acutus venom, with electrophoretic confirmation of protein integrity and preserved 740 nm fluorescence excitation. Acute toxicity assays demonstrated no significant difference in LD50 lethality between labeled and native venoms (p > 0.05). The intoxication models revealed species-specific pathophenotypes, i.e., CY7-Naja atra venom induced systemic weakness, tachypnea, and inflammatory necrosis in lung, myocardium, and liver, whereas CY7-Deinagkistrodon acutus venom provoked hemorrhagic diathesis. Both models exhibited marked leukocytosis, transaminitis, and elevated creatinine levels (p < 0.05). Fluorescence tracing uncovered distinct biodistribution kinetics:Deinagkistrodon acutus venom achieved peak organ accumulation at 3 h with rapid dissemination (24 h injection-site retention: 12.61%), contrasting withNaja atra venom’s delayed 6 h peak and prolonged renal sequestration (24 h injection-site retention: 60.9%). Target organ proteomic profiling identifiedDeinagkistrodon acutus-enriched thrombin-like enzymes and metalloproteinases in lung/liver/spleen, whileNaja atra venom predominantly accumulated renal acidic phospholipase A2 and weakly neurotoxic NNAM2.Full article
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12 pages, 438 KB  
Article
Early Botulinum Toxin Type A Injection May Improve Motor Recovery in Patients with Post-Stroke Spasticity: A Secondary Analysis from a Longitudinal Cohort Study
byAlessandro Picelli,Andrea Santamato,Michela Cosma,Alessio Baricich,Carmelo Chisari,Marzia Millevolte,Cristina Del Prete,Ilenia Mazzù,Rita Di Censo,Nicola Smania andMirko Filippetti
Toxins2025,17(11), 558;https://doi.org/10.3390/toxins17110558 - 13 Nov 2025
Abstract
Spasticity after stroke impairs motor control, delays recovery, and reduces quality of life. Botulinum toxin type A is the first-line treatment, but it is often administered in the chronic phase, potentially limiting its impact on rehabilitation. Emerging evidence suggests that earlier treatment may [...] Read more.
Spasticity after stroke impairs motor control, delays recovery, and reduces quality of life. Botulinum toxin type A is the first-line treatment, but it is often administered in the chronic phase, potentially limiting its impact on rehabilitation. Emerging evidence suggests that earlier treatment may enhance recovery, though functional benefits remain uncertain. We conducted a secondary analysis of a multicenter, open-label, longitudinal cohort study to investigate whether the timing of the first botulinum toxin type A injection influences outcomes in post-stroke patients naïve to this treatment. All participants received botulinum toxin injections combined with conventional rehabilitation. Assessments were performed at baseline and at 4, 12, and 24 weeks post-injection. The primary outcome was muscle tone; secondary outcomes included motor strength, sensorimotor recovery, and global disability. Statistical analyses used mixed-effects models and trend tests. Patients treated within 90 days of stroke onset showed greater reductions in spasticity at 4 and 12 weeks compared with later treatment. Despite having more severe baseline impairments, early treated patients demonstrated faster and more pronounced improvements in upper-limb strength, sensorimotor recovery, and global disability. Early toxin administration is associated with enhanced reduction in spasticity and improved motor recovery, particularly in patients with severe initial deficits.Full article
(This article belongs to the Special IssueBotulinum Toxin: Advancing Treatments for Spasticity)
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19 pages, 2162 KB  
Article
Optimization by the 4S Sequential Experimental Design Process of a Competitive Lateral Flow Immunoassay Device for the Detection of Aflatoxin B1
bySimone Cavalera,Sofia Stanzani,Thea Serra,Valentina Testa,Fabio Di Nardo,Claudio Baggiani andLaura Anfossi
Toxins2025,17(11), 557;https://doi.org/10.3390/toxins17110557 - 13 Nov 2025
Abstract
Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic compound produced by certain fungi (e.g.,Aspergillus flavus andAspergillus parasiticus). Rapid and ultra-sensitive detection methods for AFB1 in various commodities are in high demand. This study aimed to enhance the sensitivity of [...] Read more.
Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic compound produced by certain fungi (e.g.,Aspergillus flavus andAspergillus parasiticus). Rapid and ultra-sensitive detection methods for AFB1 in various commodities are in high demand. This study aimed to enhance the sensitivity of a competitive lateral flow immunoassay (LFIA) for AFB1 detection by leveraging a previously developed experimental design strategy, named 4S. This approach comprises four phases—START, SHIFT, SHARPEN, and STOP—and involves the analysis of two reference conditions: NEG (0 ng/mL AFB1) and POS (1 ng/mL AFB1). By generating and overlaying response surfaces, regions of optimal NEG signal and POS/NEG signal ratio (IC%) were identified. Four variables were optimized: two related to the labeled antibody (its concentration and antibody-to-label ratio) and two to the competitor antigen (its concentration and hapten-to-protein ratio). An initial design defined the parameter space, while three subsequent designs did not yield further improvements in sensitivity. A strong anti-correlation was observed between the IC% and competitor parameters. The optimized LFIA-1 exhibited enhanced sensitivity, achieving a limit of detection of 0.027 ng/mL compared to 0.1 ng/mL for the original device. Additionally, the amount of expensive antibody required for device fabrication was reduced by around a factor of four.Full article
(This article belongs to the SectionMycotoxins)
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22 pages, 2245 KB  
Review
Adsorption–Degradation Integrated Approaches to Mycotoxin Removal from Food Matrices: A Comprehensive Review
byXiyu Yang,Mingjian Yao,Wenchao Liao andXiaoyang Li
Toxins2025,17(11), 556;https://doi.org/10.3390/toxins17110556 - 12 Nov 2025
Abstract
Mycotoxin contamination is a crucial issue in food safety. However, the removal of trace amounts of mycotoxins from complex food and feed matrices without significant loss of nutritional and flavor quality remains a significant challenge. The integrated adsorption–catalysis strategy involves immobilizing catalytic modules [...] Read more.
Mycotoxin contamination is a crucial issue in food safety. However, the removal of trace amounts of mycotoxins from complex food and feed matrices without significant loss of nutritional and flavor quality remains a significant challenge. The integrated adsorption–catalysis strategy involves immobilizing catalytic modules onto adsorption materials, enabling in situ degradation while enriching the mycotoxins. This approach can significantly reduce the dosage of detoxification agents and achieve efficient removal of trace mycotoxins in food. This review provides an overview of adsorbents with enrichment capabilities and their applications in the targeted removal of mycotoxins from food. The adsorption–degradation coupled systems are categorized into the following two main types: adsorption–photocatalysis coupled systems and adsorption–biocatalysis coupled systems. The review introduces recent advances in the design of bifunctional catalysts, focusing on their synergistic mechanisms and practical applications for detoxifying various mycotoxins in food matrices. Finally, the review discusses current industrial challenges and offers insights into future directions for this field.Full article
(This article belongs to the Special IssueAdvances in Detection and Removal of Mycotoxins in Food)
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