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Integrating Telemedicine in Botulinum Toxin Type-A Treatment for Spasticity Management: Perspectives and Challenges from Italian Healthcare Professionals
Paralytic Shellfish Poisoning (PSP) Toxins in Bivalve Molluscs from Southern Italy Analysed by Liquid Chromatography Coupled with High-Resolution Mass Spectrometry (UHPLC-HRMS/MS)
The Effects of Indoxyl Sulfate in the Exacerbation of Colorectal Cancer During the Progression of Chronic Kidney Disease

Journal Description

Toxins

Toxins is an international, peer-reviewed, open access journal which provides an advanced forum for studies related to toxinology and all kinds of toxins (biotoxins) from animals, microbes and plants. Toxins is published monthly online by MDPI. The French Society on Toxinology (SFET), International Society for Mycotoxicology (ISM), Japanese Society of Mycotoxicology (JSMYCO) and European Uremic Toxins (EUTox) Work Group are affiliated with Toxins and their members receive a discount on the article processing charges.

Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, MEDLINE, PMC, Embase, CAPlus / SciFinder, AGRIS, 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 20.3 days after submission; acceptance to publication is undertaken in 2.9 days (median values for papers published in this journal in the second half of 2024).
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 6 topical sections.

Impact Factor: 3.9 (2023); 5-Year Impact Factor: 4.2 (2023)

Imprint Information Journal Flyer Open Access ISSN: 2072-6651

Latest Articles

22 pages, 6784 KiB

Open AccessArticle

Transcriptomic Characterization of the Porcine Urinary Bladder Trigone Following Intravesical Administration of Resiniferatoxin: Insights from High-Throughput Sequencing

by Ewa Lepiarczyk, Mateusz Maździarz, Łukasz Paukszto, Agnieszka Bossowska, Mariusz Majewski, Jerzy Kaleczyc, Elżbieta Łopieńska-Biernat, Łukasz Jaśkiewicz, Agnieszka Skowrońska, Mariusz T. Skowroński and Marta Majewska

Toxins 2025, 17(3), 127; https://doi.org/10.3390/toxins17030127 (registering DOI) - 9 Mar 2025

Resiniferatoxin (RTX), a potent capsaicin analog, is being investigated as a therapeutic agent for neurogenic conditions, particularly those affecting bladder control. However, the transcriptomic effects of RTX on the urinary bladder remain largely unexplored. This study aimed to characterize the transcriptomic changes in [...] Read more.

Resiniferatoxin (RTX), a potent capsaicin analog, is being investigated as a therapeutic agent for neurogenic conditions, particularly those affecting bladder control. However, the transcriptomic effects of RTX on the urinary bladder remain largely unexplored. This study aimed to characterize the transcriptomic changes in the porcine urinary bladder trigone region removed seven days post-treatment with intravesical RTX administration (500 nmol per animal in 60 mL of 5% aqueous solution of ethyl alcohol). High-throughput sequencing identified 126 differentially expressed genes (DEGs; 66 downregulated, 60 upregulated), 5 differentially expressed long non-coding RNAs (DELs), and 22 other RNAs, collectively involved in 175 gene ontology (GO) processes. Additionally, differential alternative splicing events (DASes) and single nucleotide variants (SNVs) were detected. RTX significantly modulated signaling pathways related to nerve growth and myelination. Changes in genes associated with synaptic plasticity and neuromodulation were observed, particularly within serotoninergic and cholinergic signaling. RTX altered the expression of immune-related genes, particularly those involved in chemokine signaling and immune regulation. Notably, altered gene expression patterns suggest a potential anti-cancer role for RTX. These findings provide new insights into RTX’s therapeutic effects beyond TRPV1 receptor interactions, filling a critical gap in our understanding of its molecular impact on bladder tissue. Full article

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49 pages, 2741 KiB

Open AccessReview

Health and Environmental Impacts of Cyanobacteria and Cyanotoxins from Freshwater to Seawater

by Tamara Villalobos, Benjamín Suárez-Isla and Carlos Garcia

Toxins 2025, 17(3), 126; https://doi.org/10.3390/toxins17030126 - 7 Mar 2025

Cyanobacterial harmful algal blooms (cyanoHABs) are a natural phenomenon produced mainly by the interaction between natural and anthropogenic events. CyanoHABs are characterized by the production of cyanotoxins that can have harmful effects on different species within the food web and even affect human [...] Read more.

Cyanobacterial harmful algal blooms (cyanoHABs) are a natural phenomenon produced mainly by the interaction between natural and anthropogenic events. CyanoHABs are characterized by the production of cyanotoxins that can have harmful effects on different species within the food web and even affect human health. Among the most prevalent toxin groups worldwide are microcystins (MCs), anatoxins (ATXs), cylindrospermopsins (CYNs) and nodularins (NODs), which are characterized as toxins with hepatotoxic, neurotoxic, and cytotoxic effects. This review summarizes and analyzes research on the influence of cyanoHABs, the main toxin-producing cyanobacteria and the most prevalent cyanotoxins in freshwater and marine bodies, highlighting their global occurrence, toxicology, and bioaccumulation dynamics in vectors of the food web, and the main cases of acute and chronic intoxications in humans. This review is useful for understanding the dynamics of cyanoHABs’ interaction with the ecosystem and their impact on human health, and how the implementation of a surveillance and management framework for cyanobacteria and cyanotoxins could generate vital information for stakeholders to establish health guidelines on the risks and hazards of cyanoHABs for the ecosystem and humans. Full article

(This article belongs to the Special Issue Advances in Cyanotoxins: Latest Developments in Risk Assessment)

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15 pages, 968 KiB

Open AccessArticle

Learnings from Separate Aconitum Poisonings in British Columbia and Ontario, Canada in 2022

by Lorraine McIntyre, Stefanie Georgopoulos, Dorianna Simone, Emily Newhouse, JoAnne Fernandes, David A. McVea, Arnold Fok, Ania-Maria McIntyre, Bryn Shurmer, Marie-Claude Gagnon, Michael Chan, Marina Chiaravalloti, Nikita Saha Turna, Debra Kent, Dennis Leong, Katherine Paphitis, Christina Lee and the Outbreak Investigation Teams

Toxins 2025, 17(3), 125; https://doi.org/10.3390/toxins17030125 - 7 Mar 2025

Background: Three aconitine poisoning events occurred in two Canadian provinces in 2022: one in British Columbia (BC) and two in Ontario (ON). Aconitine is a potent alkaloid found in several species of the plant Aconitum, containing cardiotoxins and neurotoxins. It is used [...] Read more.

Background: Three aconitine poisoning events occurred in two Canadian provinces in 2022: one in British Columbia (BC) and two in Ontario (ON). Aconitine is a potent alkaloid found in several species of the plant Aconitum, containing cardiotoxins and neurotoxins. It is used in traditional Chinese medicine (TCM) for pain management, and in powdered form, Aconitum is similar in appearance to sand ginger (Kaempferia galanga), which can lead to poisonings from misidentification and mislabeling. Methods: Aconitine poisoning is rare in Canada; here, we compare communications, collaborations, laboratory testing options and actions during investigations. Results: Fourteen cases occurred from the consumption of sand ginger: in BC (n = 2), purchased at an Asian health food store; in ON (n = 11), Kaempferia galanga powder (KGP) spices were used to prepare meals at a restaurant, and in one ON case, KGP was purchased. Traceback found product imported from China contained aconitine levels ranging from 1304 to 5500 ppm. Later investigations revealed mislabeling of Aconitum as KGP from the same imported lot (January 2020). Plant DNA testing found no KGP in any spice packets, including lots not linked to illness, suggestive of adulteration. Conclusion: Method development for aconitine in BC led to an improved response time for testing in ON. BC and ON updated outbreak response protocols and communications. Full article

(This article belongs to the Special Issue Plant Toxin Emergency)

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16 pages, 4035 KiB

Open AccessArticle

Abiotic Degradation of the Toxin Simplexin by Soil Collected from a Pimelea-Infested Paddock

by Zhi Hung Loh, Natasha L. Hungerford, Diane Ouwerkerk, Athol V. Klieve and Mary T. Fletcher

Toxins 2025, 17(3), 124; https://doi.org/10.3390/toxins17030124 - 6 Mar 2025

Pimelea poisoning of cattle is caused by the toxin simplexin present in native Pimelea plant species. Surface weathering and burial of Pimelea plant material under soil in Pimelea-infested pastures previously showed simplexin degradation, suggesting soil microbial metabolism and/or abiotic degradation of simplexin [...] Read more.

Pimelea poisoning of cattle is caused by the toxin simplexin present in native Pimelea plant species. Surface weathering and burial of Pimelea plant material under soil in Pimelea-infested pastures previously showed simplexin degradation, suggesting soil microbial metabolism and/or abiotic degradation of simplexin in the field. This current study investigated whether soil from a Pimelea-infested paddock was capable of simplexin degradation in the laboratory. The effects of temperature on isolated simplexin levels and simplexin levels in Pimelea plant material treated with field-collected soil, acid-washed sand or bentonite were determined. Pimelea plant material incubated in field-collected soil at 22 °C for seven days did not show any simplexin degradation. Isolated simplexin preadsorbed to field-collected soil, acid-washed sand or bentonite showed simplexin decrease after one hour of incubation at 100 °C with three breakdown products identified by UPLC-MS/MS, indicating that toxin breakdown can be a heat-induced process rather than a microbial-based metabolism. Decreased simplexin levels were observed in Pimelea plant material mixed with acid-washed sand under similar incubation conditions. Overall, the study showed the field-collected soil did not contain soil microorganisms capable of simplexin metabolism within a short period of time. However, the co-exposure to high temperature resulted in significant abiotic simplexin breakdown, without microorganism involvement, with the product structures suggesting that the degradation was a heat promoted acid hydrolysis/elimination process. Overall, this study demonstrated that simplexin breakdown in the field could be a thermal abiotic process with no indication of microbial involvement. Full article

(This article belongs to the Section Plant Toxins)

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8 pages, 411 KiB

Open AccessCommunication

Deoxynivalenol Determination Using Innovative Lateral Flow Device Technology

by Halberstadt Delphine, Giorni Paola, Barato Erica and Bertuzzi Terenzio

Toxins 2025, 17(3), 123; https://doi.org/10.3390/toxins17030123 - 5 Mar 2025

Deoxynivalenol (DON), generally the most widespread mycotoxin in wheat, is regulated by the EU regulation in cereals and cereal-derived products. Its presence can be detected by chromatographic or rapid methods; the latter technique is generally used in control analysis, fulfilling the needs of [...] Read more.

Deoxynivalenol (DON), generally the most widespread mycotoxin in wheat, is regulated by the EU regulation in cereals and cereal-derived products. Its presence can be detected by chromatographic or rapid methods; the latter technique is generally used in control analysis, fulfilling the needs of the stakeholders of the wheat grain chain. Lateral flow strips are often used for the rapid detection of different mycotoxins in several agricultural products; regarding DON determination, different lateral flow immunochromatography strips are currently available, also providing quantitative results. The purpose of this work was to evaluate the accuracy of an innovative lateral flow device coupled to a bench top device, following a digital approach. The proposed method was compared to an LC-MS/MS method, analyzing 50 naturally contaminated wheat samples. The results obtained using the two methods were very similar and, applying a paired t-test, the mean difference between measurements resulted not significantly different (α = 0.003). The correlation between the results showed a slope of the line close to 1 (m = 0.9904) and a regression coefficient (r) of 0.9968. Full article

(This article belongs to the Special Issue Analytical Novelties and Challenges for the Detection of Natural Toxins)

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12 pages, 269 KiB

Open AccessReview

Review: Botulinum Toxin for Treatment of Focal Limb Dystonia

by Emma H. Kaplan, Michele Vecchio and David M. Simpson

Toxins 2025, 17(3), 122; https://doi.org/10.3390/toxins17030122 - 4 Mar 2025

Focal limb dystonias (FLDs) are abnormal postures and muscle contractions in an arm or leg that can occur in the setting of specific activities or without any stimulus. This pathology can have a profound impact on quality of life and potentially limit work [...] Read more.

Focal limb dystonias (FLDs) are abnormal postures and muscle contractions in an arm or leg that can occur in the setting of specific activities or without any stimulus. This pathology can have a profound impact on quality of life and potentially limit work in those whose dystonias are brought on by activities related to their occupations. Botulinum toxin (BoNT) is approved for use in the United States by the Food and Drug Administration for several indications, including cervical dystonia and blepharospasm, but not for FLD. Despite this limitation, BoNT is frequently used clinically for FLD, generally with individualized dosing based on patient need and clinician expertise. Various methods exist for targeting treatment to the specific affected muscles and assessing the benefit of treatment. Small clinical trials have demonstrated the efficacy of BoNT, but larger controlled studies are needed. Full article

(This article belongs to the Special Issue Botulinum Toxins Injections for Hypertonic Limbs—Is It So Effective?)

18 pages, 7929 KiB

Open AccessCase Report

The Relationship Between Stiff Knee Gait Runner’s Dystonia and Musculoskeletal Knee Pathology: A Case Series

by Jared A. Stowers, Derek S. Day, Steven Jow, Sarah Heins, Euan Forrest, Yonathan M. Assefa, Paige M. Lind, Afreen Mushtaheed, Frances T. Sheehan and Katharine E. Alter

Toxins 2025, 17(3), 121; https://doi.org/10.3390/toxins17030121 - 3 Mar 2025

Background: Runner’s dystonia (RD), a rare task-specific lower-limb dystonia affecting high-mileage runners, presents as abnormal lower-extremity muscle contractions during running. Treatment of RD is challenging and often confounded by significant diagnostic delays due to overlapping symptomatology with other conditions. This case series examines [...] Read more.

Background: Runner’s dystonia (RD), a rare task-specific lower-limb dystonia affecting high-mileage runners, presents as abnormal lower-extremity muscle contractions during running. Treatment of RD is challenging and often confounded by significant diagnostic delays due to overlapping symptomatology with other conditions. This case series examines the relationship between stiff knee gait RD and musculoskeletal (MSK) knee pathology. Methods: Eight RD cases, evaluated at the NIH Movement Disorders Clinic since 2018, were retrospectively reviewed. Patients underwent neurological, biomechanical, and MSK evaluations, including 3D motion analysis, surface electromyography, and knee ultrasound. Therapeutic interventions, including botulinum neurotoxin (BoNT) injections, were assessed. Results: Seven patients demonstrated stiff knee gait subtypes, with all having ipsilateral and/or contralateral knee effusions or tendinopathies. Three patients who received MSK interventions (e.g., aspiration, corticosteroid injections) combined with BoNT therapy experienced significant symptom improvement. One patient with isolated foot dystonia displayed different biomechanical patterns without knee pathology. Conclusions: RD patients with stiff knee gait often exhibit knee pathology, most likely due to altered biomechanics and running history. Addressing both issues is essential for optimizing treatment outcomes, reducing pain, and improving function, especially since pain can trigger dystonia. Future research should determine the ideal sequence of interventions for RD patients with MSK issues to develop effective, personalized treatment algorithms. Full article

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14 pages, 843 KiB

Open AccessArticle

An Unusual Outbreak of Ochratoxicosis Associated with Trigonella foenum-graecum Ingestion in Ruminants from Different Farms of Sicily

by Maria Rizzo, Patrizia Licata, Pietro Paolo Niutta, Michela Pugliese, Vito Macaluso, Giovanna Lucrezia Costa, Giuseppe Bruschetta and Fabio Bruno

Toxins 2025, 17(3), 120; https://doi.org/10.3390/toxins17030120 - 2 Mar 2025

Trigonella foenum-graecum is a widely cultivated legume in Mediterranean regions, and it is used for human and animal consumption, as well as for medical purposes. High temperatures and abundant rainfall during the spring season in Sicily favor the formation of an environment suitable [...] Read more.

Trigonella foenum-graecum is a widely cultivated legume in Mediterranean regions, and it is used for human and animal consumption, as well as for medical purposes. High temperatures and abundant rainfall during the spring season in Sicily favor the formation of an environment suitable for the growth and proliferation of fungi with the production of mycotoxins. In this study, ochratoxin A, aflatoxin, deoxynivalenol, zearalenone, fumonisin, and T-2 toxin concentrations in Trigonella foenum-graecum were determined in feed administered to ruminants and also in blood samples from cattle and sheep in order to evaluate the toxicity correlated to the possible presence of these mycotoxins based on the clinical signs observed in the animals. Analyses of mycotoxins in fenugreek and blood samples were conducted using the enzyme immunoassay KIT. Five extensive farms sited in the northwest of the Sicily region, with a total of 90 intoxicated animals, reported a concomitant unusual outbreak of neurological disorders. Decreased spinal reflex responses, postural abnormalities associated with weakness or recumbency, and hyperesthesia of the limbs suggested a problem regarding the peripheral nervous system. The mortality rate recorded was very high, even reaching 100% of the intoxicated animals. OTA intoxication in Sicilian ruminants represents an important warning on the vulnerability of farms to mycotoxin contamination and underlines the importance of preventive measures and monitoring in animal health management. Full article

(This article belongs to the Section Mycotoxins)

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18 pages, 3605 KiB

Open AccessArticle

Proteo-Transcriptomic Analysis of the Venom Gland of the Cone Snail Cylinder canonicus Reveals the Origin of the Predatory-Evoked Venom

by Zahrmina Ratibou, Anicet E. T. Ebou, Claudia Bich, Fabrice Saintmont, Gilles Valette, Guillaume Cazals, Dominique K. Koua, Nicolas Inguimbert and Sébastien Dutertre

Toxins 2025, 17(3), 119; https://doi.org/10.3390/toxins17030119 - 2 Mar 2025

Cone snails are carnivorous marine predators that prey on mollusks, worms, or fish. They purposefully inject a highly diversified and peptide-rich venom, which can vary according to the predatory or defensive intended use. Previous studies have shown some correlations between the predation- and [...] Read more.

Cone snails are carnivorous marine predators that prey on mollusks, worms, or fish. They purposefully inject a highly diversified and peptide-rich venom, which can vary according to the predatory or defensive intended use. Previous studies have shown some correlations between the predation- and defense-evoked venoms and specific sections of the venom gland. In this study, we focus on the characterization of the venom of Cylinder canonicus, a molluscivorous species collected from Mayotte Island. Integrated proteomics and transcriptomics studies allowed for the identification of 108 conotoxin sequences from 24 gene superfamilies, with the most represented sequences belonging to the O1, O2, M, and conkunitzin superfamilies. A comparison of the predatory injected venom and the distal, central, and proximal sections of the venom duct suggests mostly distal origin. Identified conotoxins will contribute to a better understanding of venom–ecology relationships in cone snails and provide a novel resource for potential drug discovery. Full article

(This article belongs to the Special Issue Conotoxins: Evolution, Classifications and Targets)

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27 pages, 7555 KiB

Open AccessArticle

Cylindracin, a Fruiting Body-Specific Protein of Cyclocybe cylindracea, Represses the Egg-Laying and Development of Caenorhabditis elegans and Drosophila melanogaster

by Yamato Kuratani, Akira Matsumoto, Ayako Shigenaga, Koji Miyahara, Keisuke Ekino, Noriaki Saigusa, Hiroto Ohta, Makoto Iwata and Shoji Ando

Toxins 2025, 17(3), 118; https://doi.org/10.3390/toxins17030118 - 1 Mar 2025

Mushrooms are a valuable source of bioactive compounds to develop efficient, secure medicines and environmentally friendly agrochemicals. Cylindracin is a small cysteine-rich protein that is specifically expressed in the immature fruiting body of the edible mushroom Cyclocybe cylindracea. Recombinant protein (rCYL), comprising [...] Read more.

Mushrooms are a valuable source of bioactive compounds to develop efficient, secure medicines and environmentally friendly agrochemicals. Cylindracin is a small cysteine-rich protein that is specifically expressed in the immature fruiting body of the edible mushroom Cyclocybe cylindracea. Recombinant protein (rCYL), comprising the C-terminal cysteine-rich domain of cylindracin, inhibits the hyphal growth and conidiogenesis of filamentous fungi. Here, we show that rCYL represses the egg-laying and development of Caenorhabditis elegans and Drosophila melanogaster. The feeding of rCYL at 16 µM reduced the body volume of C. elegans larvae to approximately 60% when compared to the control. At the same concentration, rCYL repressed the frequencies of pupation and emergence of D. melanogaster to 74% and 40%, respectively, when compared to the control. In virgin adult flies, feeding of rCYL at 47 µM substantially repressed the frequency of egg-laying, and the pupation and emergence of the next generation, especially for females. These inhibitory effects of rCYL gradually disappeared after ceasing the ingestion of rCYL. The use of fluorescence-labeled rCYL revealed that the protein accumulates specifically at the pharynx cuticles of C. elegans. In D. melanogaster, fluorescence-labeled rCYL was detected primarily in the midguts and to a lesser degree in the hindguts, ovaries, testes, and malpighian tubules. rCYL was stable against trypsin, chymotrypsin, and pepsin, whereas it did not inhibit proteolytic and glycolytic enzymes in vitro. rCYL oligomerized and formed amyloid-like aggregates through the binding to heparin and heparan sulfate in vitro. These results suggest that rCYL has potential as a new biocontrol agent against pests. Full article

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25 pages, 2471 KiB

Open AccessArticle

Pain Reduction with Repeated Injections of Botulinum Toxin A in Upper Limb Spasticity: A Longitudinal Analysis from the ULIS-III Study

by Lynne Turner-Stokes, Khan Buchwald, Stephen A. Ashford, Klemens Fheodoroff, Jorge Jacinto, Ajit Narayanan and Richard J. Siegert

Toxins 2025, 17(3), 117; https://doi.org/10.3390/toxins17030117 - 1 Mar 2025

Pain reduction is a common goal of the treatment of upper limb spasticity with botulinum toxin (BoNT-A). ULIS-III was a large international, observational, longitudinal study (N = 953) conducted in real-life clinical practice over two years. In this secondary post hoc analysis, we [...] Read more.

Pain reduction is a common goal of the treatment of upper limb spasticity with botulinum toxin (BoNT-A). ULIS-III was a large international, observational, longitudinal study (N = 953) conducted in real-life clinical practice over two years. In this secondary post hoc analysis, we examine whether goals for pain reduction were met over repeated injection cycles. We report serial changes in pain severity and explore predictors of pain reduction and injection frequency. Patients were selected if pain reduction was a primary/secondary goal for at least one cycle (n = 438/953). They were assessed at the start and end of each cycle using the goal attainment T-score alongside a self-report of pain severity (range 0–10). Across all cycles, pain-related goals were set for 1189/1838 injections (64.7%) and were achieved in 839 (70.6%). Patients continued to show a significant reduction in pain (p < 0.001) for each injection up to seven cycles, with some cumulative benefit (p < 0.001). Those requiring more frequent injections tended to have higher starting pain scores and a smaller reduction in pain score, but these differences were not significant when other covariates (age, previous injection history, time since onset, severity and distribution of spasticity) were taken into account (p > 0.713). Conclusion: Repeated BoNT-A administration continued to result in a significant reduction in upper limb spasticity-related pain, regardless of patient-related factors. Full article

(This article belongs to the Section Bacterial Toxins)

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40 pages, 2108 KiB

Open AccessArticle

Multi-Mycotoxin Contamination of Aquaculture Feed: A Global Survey

by Christiane Gruber-Dorninger, Anneliese Müller and Roy Rosen

Toxins 2025, 17(3), 116; https://doi.org/10.3390/toxins17030116 - 1 Mar 2025

Plant-based materials are increasingly being used as ingredients of aquaculture feed. These materials are prone to mycotoxin contamination, as mycotoxigenic fungi infest crop plants in the field and agricultural products during storage. As mycotoxins can cause toxic effects in aquatic animals, their occurrence [...] Read more.

Plant-based materials are increasingly being used as ingredients of aquaculture feed. These materials are prone to mycotoxin contamination, as mycotoxigenic fungi infest crop plants in the field and agricultural products during storage. As mycotoxins can cause toxic effects in aquatic animals, their occurrence in feedstuffs should be monitored. To this end, we performed an extensive global survey of mycotoxin contamination in aquaculture feed and plant-based feed raw materials. We collected samples of compound feed for fish (n = 226) and shrimps (n = 61), maize (n = 3448), maize DDGS (n = 149), wheat (n = 1578), soybean (n = 428), and rice (n = 65). We analyzed concentrations of 51 mycotoxins, emerging mycotoxins, masked mycotoxins, and mycotoxin metabolites. Mycotoxins were almost ubiquitously present in compound feed, as >90% of samples were contaminated with at least one mycotoxin. Feed raw materials exhibited distinct mycotoxin occurrence patterns consistent with known susceptibility to fungal pathogens and with their production process. Unsafe concentrations of aflatoxin B₁ exceeding the EU maximum level were detected in 7.2% of fish feed samples. While most feedstuffs complied with EU guidance values for deoxynivalenol, zearalenone, and fumonisins, a comparison of detected concentrations with dietary concentrations reported to cause adverse effects in fish and shrimps in published studies indicated that significant fractions of samples contained potentially harmful levels of these mycotoxins. In addition to regulated mycotoxins, several emerging mycotoxins (e.g., enniatins, beauvericin, alternariol, moniliformin) were prevalent. Feed was frequently co-contaminated with multiple mycotoxins indicating a risk of combined effects. In conclusion, mycotoxin contamination was common in aquaculture feed and fractions of samples were contaminated with mycotoxin levels known to exert adverse effects in aquaculture species. Results of this survey highlight the necessity for targeted studies on the effects of frequently detected mycotoxin mixtures and emerging mycotoxins in fish and shrimp. Full article

(This article belongs to the Section Mycotoxins)

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20 pages, 869 KiB

Open AccessReview

Human Deaths Related to Oleander Poisoning: A Review of the Literature

by Matteo Antonio Sacco, Saverio Gualtieri, Aurora Princi, Alessandro Pasquale Tarallo, Maria Cristina Verrina, Lucia Tarda, Luca Calanna, Santo Gratteri and Isabella Aquila

Toxins 2025, 17(3), 115; https://doi.org/10.3390/toxins17030115 - 1 Mar 2025

Oleander poisoning, resulting from the ingestion of Nerium oleander or Thevetia peruviana, is a serious toxicological issue in various parts of the world, particularly in regions where these plants grow abundantly and are easily accessible. Oleander contains potent cardiac glycosides, such as oleandrin [...] Read more.

Oleander poisoning, resulting from the ingestion of Nerium oleander or Thevetia peruviana, is a serious toxicological issue in various parts of the world, particularly in regions where these plants grow abundantly and are easily accessible. Oleander contains potent cardiac glycosides, such as oleandrin and thevetin, which exert powerful effects on the cardiovascular system, leading to symptoms ranging from nausea and abdominal pain to severe arrhythmias and sudden cardiac death. This review summarizes the existing literature on the epidemiology, clinical features, pathophysiology, and challenges in treatment management associated with oleander poisoning. While supportive care, gastric decontamination, and the administration of digoxin-specific Fab antibody fragments (Digifab) are essential therapeutic measures, limited access to Digifab, delays in intervention, and insufficient supportive care practices remain significant complicating factors. Particular attention is given to findings from autopsy reports, which provide critical insights into the pathophysiological effects of oleander toxins and help bridge gaps in understanding fatal cases. This review acknowledges key limitations, particularly the scarcity of English-language publications, which restricts input from regions such as southern Asia and the Mediterranean—areas where oleander-related poisoning, especially in cases of intentional self-harm, is more prevalent. Additionally, this review highlights the socio-cultural dimensions of oleander ingestion, often linked to intentional self-poisoning, and emphasizes the need for enhanced preventive measures and public education. Future research efforts should prioritize addressing these gaps through autopsy-based studies and the development of more accessible and effective antidotes, which are essential to mitigate the global health burden of oleander-related mortality. Full article

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26 pages, 1953 KiB

Open AccessReview

Short Peptides from Asian Scorpions: Bioactive Molecules with Promising Therapeutic Potential

by Kaiyun Xin, Ruize Sun, Wanyang Xiao, Weijie Lu, Chenhui Sun, Jietao Lou, Yanyan Xu, Tianbao Chen, Di Wu and Yitian Gao

Toxins 2025, 17(3), 114; https://doi.org/10.3390/toxins17030114 - 28 Feb 2025

Scorpion venom peptides, particularly those derived from Asian species, have garnered significant attention, offering therapeutic potential in pain management, cancer, anticoagulation, and infectious diseases. This review provides a comprehensive analysis of scorpion venom peptides, focusing on their roles as voltage-gated sodium (Nav), potassium [...] Read more.

Scorpion venom peptides, particularly those derived from Asian species, have garnered significant attention, offering therapeutic potential in pain management, cancer, anticoagulation, and infectious diseases. This review provides a comprehensive analysis of scorpion venom peptides, focusing on their roles as voltage-gated sodium (Nav), potassium (Kv), and calcium (Cav) channel modulators. It analyzed Nav1.7 inhibition for analgesia, Kv1.3 blockade for anticancer activity, and membrane disruption for antimicrobial effects. While the low targeting specificity and high toxicity of some scorpion venom peptides pose challenges to their clinical application, recent research has made strides in overcoming these limitations. This review summarizes the latest progress in scorpion venom peptide research, discussing their mechanisms of action, therapeutic potential, and challenges in clinical translation. This work aims to provide new insights and directions for the development of novel therapeutic drugs. Full article

(This article belongs to the Special Issue Animals Venom in Drug Discovery: A Valuable Therapeutic Tool)

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12 pages, 1271 KiB

Open AccessArticle

Susceptibility and Mechanism of Aflatoxin Contamination of Ziziphus jujuba var. spinosa

by Abdelrahman Elamin and Shohei Sakuda

Toxins 2025, 17(3), 113; https://doi.org/10.3390/toxins17030113 - 27 Feb 2025

The susceptibility and mechanism of aflatoxin (AF) contamination in Ziziphus jujuba var. spinosa, whose seeds are important for medicinal use, were evaluated in this study. First, the susceptibility of intact fruits, classified into four maturity groups, to AF accumulation was assessed through [...] Read more.

The susceptibility and mechanism of aflatoxin (AF) contamination in Ziziphus jujuba var. spinosa, whose seeds are important for medicinal use, were evaluated in this study. First, the susceptibility of intact fruits, classified into four maturity groups, to AF accumulation was assessed through artificial contamination with an aflatoxigenic Aspergillus flavus strain. AF analysis revealed that mid-mature fruits were highly susceptible to AF contamination. Next, AF accumulation in seed parts was examined by artificially inoculating A. flavus on intact fruits, showing AF presence in seeds after 30 days of incubation. The susceptibility of jujube kernels to AF accumulation in seed parts was then studied. The artificial inoculation of A. flavus on kernels, classified into three groups based on the pedicel condition, showed no correlation between AF contamination and the pedicel condition, with large fluctuations within each group. Finally, the effect of the hilar region morphology on AF contamination in seeds was investigated. The microscopic investigation of artificially contaminated seeds and AF quantification revealed that variations in AF concentration were linked to differences in the hilar region morphology. Full article

(This article belongs to the Special Issue Aspergillus flavus and Aflatoxins (Volume III))

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18 pages, 3357 KiB

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Structurally Similar Mycotoxins Aflatoxin B₁ and Sterigmatocystin Trigger Different and Distinctive High-Resolution Mutational Spectra in Mammalian Cells

by Pennapa Thongararm, Marisa Chancharoen, Nutchapong Suwanwong, Somsak Ruchirawat, Mathuros Ruchirawat, Bogdan I. Fedeles, Robert G. Croy and John M. Essigmann

Toxins 2025, 17(3), 112; https://doi.org/10.3390/toxins17030112 - 27 Feb 2025

Aflatoxin B₁ (AFB₁) and sterigmatocystin (ST) are mycotoxins that pose significant threats to human and animal health owing to their mutagenic, carcinogenic, and toxic properties. They are structurally similar and widely believed to exert their biological effects via the generation [...] Read more.

Aflatoxin B₁ (AFB₁) and sterigmatocystin (ST) are mycotoxins that pose significant threats to human and animal health owing to their mutagenic, carcinogenic, and toxic properties. They are structurally similar and widely believed to exert their biological effects via the generation of DNA-damaging epoxides at their respective terminal furan rings. Despite structural identity in the warhead portion of each toxin, this work shows that distal parts of each molecule are responsible for the distinctive mutational fingerprints seen in gptΔ C57BL/6J mouse embryo fibroblasts (MEFs). The two toxins differ structurally in the puckered cyclopentenone ring of AFB₁ and in the planar xanthone functionality of ST. While both toxins mainly induce GC→TA mutations, the aforementioned differences in structure apparently trigger unique patterns of mutations, as revealed by high-resolution duplex sequencing of MEF genomes. AFB₁ is more mutagenic than ST and displays its transversion mutations in a pattern with primary and secondary hotspots (underscored) in 5′-CGC-3′ and 5′-CGG-3′ contexts, respectively. ST displays a modest 5′-CGG-3′ hotspot while its other GC→TA transversions are more uniformly distributed in a pattern resembling established oxidative stress mutational spectra. This research delineates the mutational spectra of AFB₁ and ST, establishing these patterns as possible early-onset biomarkers of exposure. Full article

(This article belongs to the Special Issue 65 Years On from Aflatoxin Discovery—a Themed Issue in Honor of Professor John D. Groopman)

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17 pages, 6988 KiB

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Structural and Energetic Evidence Supports the Non-Covalent Phosphate Cyclization by the Class II Phospholipase D from Loxosceles intermedia

by Carolina Gismene, José Fernando Ruggiero Bachega, Daniel Z. Doherty, Silvio Sanches Veiga, Raghuvir K. Arni and Jorge Enrique Hernández González

Toxins 2025, 17(3), 111; https://doi.org/10.3390/toxins17030111 - 27 Feb 2025

Phospholipase D (PLD) enzymes from Loxosceles spider venom mediate envenomation pathology by cleaving phospholipid headgroups. We revisited the crystal structure of Loxosceles intermedia PLD (PDB: 3RLH) to evaluate two alternative mechanisms—covalent and non-covalent—for headgroup cleavage. The covalent mechanism involves a nucleophilic attack on [...] Read more.

Phospholipase D (PLD) enzymes from Loxosceles spider venom mediate envenomation pathology by cleaving phospholipid headgroups. We revisited the crystal structure of Loxosceles intermedia PLD (PDB: 3RLH) to evaluate two alternative mechanisms—covalent and non-covalent—for headgroup cleavage. The covalent mechanism involves a nucleophilic attack on the substrate’s P atom by catalytic histidine, forming a phosphohistidine intermediate. It was originally suggested that this intermediate hydrolyzes, leading to linear phosphates. The non-covalent mechanism relies on the substrate’s hydroxyl group performing an intramolecular attack on the P atom, thereby generating a cyclic phosphate. Structural refinement of the crystal structure revealed a cyclic phosphate bound at the active site, replacing previously assigned PEG molecules. This cyclic product, stabilized by His12, His47, and Mg²⁺, provides structural evidence that supports phosphate cyclization. The results of computational analyses, including molecular dynamics and quantum mechanics/molecular mechanics simulations, further support the non-covalent mechanism as the energetically preferred pathway, with a significantly lower activation barrier. Our findings highlight the role of substrate orientation and of the catalytic His residues in transphosphatidylation, advancing our understanding of PLD enzymology and providing insights for the design of inhibitors against Loxosceles envenomation. Full article

(This article belongs to the Special Issue From Animal Venoms to Solutions: In Honor of Professor Lourival D. Possani on the Occasion of His 85th Birthday)

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11 pages, 932 KiB

Open AccessArticle

Resilience to Climate Change by Biocontrol Yeasts Against Ochratoxin A Production in Robusta Coffee

by Claudia López-Rodríguez, Carol Verheecke-Vaessen, Caroline Strub, Angélique Fontana, Tagro Guehi, Sabine Schorr-Galindo and Angel Medina

Toxins 2025, 17(3), 110; https://doi.org/10.3390/toxins17030110 - 27 Feb 2025

Aspergillus carbonarius is the main producer of Ochratoxin A (OTA) in coffee. In the last few years, there has been an increasing interest in using yeast isolates as Biocontrol Agents to prevent OTA production in coffee cherries during the primary postharvest processing. Little [...] Read more.

Aspergillus carbonarius is the main producer of Ochratoxin A (OTA) in coffee. In the last few years, there has been an increasing interest in using yeast isolates as Biocontrol Agents to prevent OTA production in coffee cherries during the primary postharvest processing. Little is known about how climate change abiotic conditions of increased temperature (+2–4 °C), elevated CO₂ (existing levels of 400 vs. 1000 ppm), and increased drought stress will impact biocontrol resilience. This study examined the effect of a three-way interaction between temperature (27, 30, and 33 °C) x water activity (a_w) (0.90 and 0.95 a_w) x CO₂ level (400 vs. 1000 ppm) on the growth and OTA production of A. carbonarius and the resilience of three yeast strains’ biocontrol capacity on fresh coffee cherries. High a_w (0.95), CO₂, and temperature levels increased the production of OTA by A. carbonarius. All the yeast biocontrol strains significantly reduced A. carbonarius growth by at least 20% and OTA production by up to 85%. From the three strains used, the Meyerozyma caribbica strain (Y4) showed the best resilience to climate change, since it reduced both growth (50%) and OTA production (70%) under future scenarios of CO₂ and a_w at all temperatures tested, and should be the one selected for pilot scale experiments in Ivory Coast. Full article

(This article belongs to the Collection Impact of Climate Change on Fungal Population and Mycotoxins)

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15 pages, 2412 KiB

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Tolerance to a Diet of Toxic Microcystis aeruginosa in Caenorhabditis elegans

by Jordan Balson, Jeffrey R. Boudreau, Ian D. Chin-Sang, Yuxiang Wang and Daniel D. Lefebvre

Toxins 2025, 17(3), 109; https://doi.org/10.3390/toxins17030109 - 27 Feb 2025

Reported incidences of cyanobacterial harmful algal blooms (CHABs) are increasing across the world due to climate change and nutrient loading, dominating freshwater ecosystems and producing dangerous cyanotoxins that cause ecological damage. Microcystis aeruginosa is one of the most common species of cyanobacteria; it [...] Read more.

Reported incidences of cyanobacterial harmful algal blooms (CHABs) are increasing across the world due to climate change and nutrient loading, dominating freshwater ecosystems and producing dangerous cyanotoxins that cause ecological damage. Microcystis aeruginosa is one of the most common species of cyanobacteria; it produces hepatotoxic and neurotoxic microcystin-LR. The ecological and human impact of algal blooms is immense, and traditional CHAB remediation methods are not always adequate in eutrophic regions such as Lake Erie in North America. As a result, a proactive, targeted approach is needed to bioremediate cyanobacteria in their pre-colonial stages. Nematodes, such as the model organism Caenorhabditis elegans, are potential candidates for bioremediating cyanobacteria such as M. aeruginosa. C. elegans have metabolic pathways that could detoxify microcystin-LR and enable tolerance to cyanobacteria in nature. We analyzed C. elegans health and fat accumulation on a diet of toxic M. aeruginosa and found that C. elegans can ingest, digest, metabolize, and survive off of this diet. The mean lifespans of the worm populations were only slightly different at 20.68 ± 0.35 (mean ± S.E.M) and 17.89 ± 0.40 when fed E. coli and toxic M. aeruginosa, respectively. In addition, a diet of toxic M. aeruginosa compared to E. coli did not have any significant impact on C. elegans pharyngeal pumping (304.2 ± 9.3 versus 330.0 ± 10.4 pumps/min), dauer response (86.3 ± 1.0 versus 83.65 ± 1.0% in dauer), mobility (209.25 ± 7.0 versus 210.15 ± 4.4 thrashes/min), or SKN-1 expression based on SKN1::GFP fluorescence measurements. Overall, a diet of toxic M. aeruginosa was able to sustain C. elegans development, and C. elegans was tolerant of it. These results suggest that C. elegans and similar nematodes could be viable candidates for cyanobacterial bioremediation. Full article

(This article belongs to the Special Issue Unraveling the Environmental Threat of Harmful Algae Blooms (HABs): Causes and Impacts)

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17 pages, 7243 KiB

Open AccessArticle

N-Glycosylation Profile of Abrin Certified EU Reference Material

by Roland Josuran, Andreas Wenger, Sylvia Worbs, Bettina Kampa, Andreas Rummel, Brigitte G. Dorner and Sabina Gerber

Toxins 2025, 17(3), 108; https://doi.org/10.3390/toxins17030108 - 26 Feb 2025

Abrin is a highly toxic plant protein encompassing four isoforms, abrin-a, -b, -c and -d. An abrin reference material was isolated from Abrus precatorius and certified (EURM-113) by the EuroBioTox consortium. Here, we present a detailed characterisation of the N-glycosylation profile of [...] Read more.

Abrin is a highly toxic plant protein encompassing four isoforms, abrin-a, -b, -c and -d. An abrin reference material was isolated from Abrus precatorius and certified (EURM-113) by the EuroBioTox consortium. Here, we present a detailed characterisation of the N-glycosylation profile of EURM-113. The monosaccharide composition of the N-glycans was determined and quantified. Release of the N-glycans yielded 13 different partially xylosylated, oligomannosidic and paucimannosidic glycan structures. Two N-glycans were found at N82 and N110 of the abrin-b A-chain and another two at N100 and N140 of the B-chains. The N-glycosylation sites N200 in the A-chain and N141 in the B-chain were non-glycosylated. Whereas N82 and N110 of abrin-b comprised paucimannosidic glycans, N100 and N140 of the B-chains revealed oligomannosidic N-glycans. Xylose was absent in the glycans at N100 but was present in about half of the glycans at N140. Hence, this study revealed substantially different types of glycan structures within the B-chains compared to the abrin-b A-chain. Furthermore, the most C-terminal N-glycosylation site in the A-chain was found to be non-glycosylated in all abrin isoforms detected. Additionally, the establishment of the N-glycosylation profile of the abrin reference material led to the identification of the abrin isoforms -a, -b and -c. In conclusion, the abrin N-glycosylation profile is highly similar to the one of ricin and yields high analytical value to be further exploited as a fingerprint in forensic investigations to uncover toxin production or toxin provenance. Full article

(This article belongs to the Special Issue The EuroBioTox Project: Progress Towards the Detection and Identification of Biotoxins via Standardization, Training, and Method Evaluation)

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