Rethinking Botox Aftercare: 5000 Patients with a 10-Minute Protocol
Extensive Variation in Thermal Responses and Toxin Content Among 40 Strains of the Cold-Water Diatom Pseudo-nitzschia seriata

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 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 6 topical sections.

Impact Factor: 4.0 (2024); 5-Year Impact Factor: 4.5 (2024)

Imprint Information Journal Flyer Open Access ISSN: 2072-6651

Latest Articles

22 pages, 804 KiB

Open AccessArticle

Prevalence and Risk Assessment of Multiple Mycotoxins in Durum Wheat from Fields Under Different Agricultural Practices in Tunisia

by Marwa Hassine, Khouloud Ben Hassouna, Salma Tissaoui, Mokhtar Baraket, Amine Slim, Olfa Ayed Slama, Hajer Slim Amara, Ahmed Al-Amiery, Noelia Pallarés, Houda Berrada, Samir Abbès and Jalila Ben Salah-Abbès

Toxins 2025, 17(8), 410; https://doi.org/10.3390/toxins17080410 (registering DOI) - 14 Aug 2025

Abstract

Mycotoxin contamination in wheat, a staple food critical to human nutrition, poses significant public health concerns. This study investigated the natural occurrence of 17 mycotoxins in Tunisian durum wheat, assessed the influence of soil tillage practices on mycotoxin contamination, and performed an associated exposure risk assessment. A total of 167 wheat samples were randomly collected over two years (2021 and 2022) from fields managed under conventional tillage (CT) and no-tillage (NT) systems during both pre- and post-harvest periods. Mycotoxins were extracted using the QuEChERS method and quantified via UHPLC-MS/MS. The results demonstrated contamination by ZEN, DON, OTA, ENA1, ENB, and ENB1. Among regulated mycotoxins, OTA was the most prevalent, detected in 68 out of 167 samples with a mean concentration of 1.85 µg/kg. ZEN was the most abundant, detected in 65 samples with a mean concentration of 26.85 µg/kg, while DON was less frequently detected in 62 samples with a mean concentration of 0.68 µg/kg. Regarding emerging mycotoxins, ENB was the most prevalent and abundant, found in 51 samples with a mean concentration of 10.13 µg/kg; ENB1 and ENA1 were detected in 20 and 10 samples, with mean concentrations of 3.38 µg/kg and 1.69 µg/kg, respectively. Furthermore, mycotoxin concentrations varied according to agricultural practices. DON, ZEN, ENA1, ENB, and ENB1 showed higher frequencies and concentrations (ranging from 0.08 to 210.11 µg/kg) in samples collected during the 2021 pre-harvest period from NT fields. In contrast, OTA exhibited greater prevalence and higher concentrations (ranging from 2.33 to 9.78 µ/kg) in samples collected during the 2022 post-harvest period from CT fields. The Estimated Daily Intake (EDI) of mycotoxins by Tunisian adults was calculated based on contamination levels in raw durum wheat from fields under NT and CT practices, resulting in the following values (ng/kg bw/day), with the first value corresponding to NT samples and the second to CT samples: OTA (17.3; 20.8), ZEN (466.3; 194.0), DON (8.0; 7.56), ENA1 (4.30; 18.85), ENB (105.17; 121.08), and ENB1 (49.91; 40.91). Both the Margin of Exposure (MOE) values for OTA and the Hazard Quotients (HQ) for ZEN and DON exceeded established safety thresholds, indicating potential health risks for Tunisian adults. These findings highlight the urgent need to implement stricter mycotoxin regulations in Tunisia and enhance surveillance systems. Further research is warranted to elucidate the mechanisms by which soil tillage practices influence mycotoxin contamination and to develop targeted mitigation strategies to ensure food safety. Full article

(This article belongs to the Special Issue Strategies for Mitigating Mycotoxin Contamination in Food and Feed)

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16 pages, 1011 KiB

Open AccessArticle

Effect of Dietary Difructose Anhydride III Supplementation on the Metabolic Profile of Japanese Black Breeding Herds with Low-Level Chronic Exposure to Zearalenone in the Dietary Feed

by Topas Wicaksono Priyo, Jr., Naoya Sasazaki, Katsuki Toda, Hiroshi Hasunuma, Daisaku Matsumoto, Emiko Kokushi, Seiichi Uno, Osamu Yamato, Takeshi Obi, Urara Shinya, Oky Setyo Widodo, Yasuho Taura, Tetsushi Ono, Masayasu Taniguchi and Mitsuhiro Takagi

Toxins 2025, 17(8), 409; https://doi.org/10.3390/toxins17080409 (registering DOI) - 14 Aug 2025

Abstract

Mycotoxin contamination in animal feed can cause acute or chronic adverse effects on growth, productivity, and immune function in livestock. This study aimed to evaluate the impact of difructose anhydride III (DFA III) supplementation on serum biochemical parameters and intestinal environment in Japanese Black (JB) breeding cows under low-level chronic dietary exposure to zearalenone (ZEN). Using urinary ZEN concentration as an indicator of exposure, 25 JB cows were selected from a breeding farm with confirmed natural feed contamination. Blood samples were collected before DFA III supplementation (day 0), and on days 20 and 40 post-supplementation. Serum biochemical parameters and short-chain fatty acid concentrations were measured. During the studies, dietary ZEN concentration increased, yet improvements were observed in liver function, nutritional status, immune response, and inflammatory markers. Notably, serum butyrate concentration significantly increased following DFA III administration. These findings suggest that DFA III may positively influence intestinal microflora and enhance intestinal barrier function, which could contribute to improved health and nutritional status in cattle exposed to low-level chronic dietary ZEN contamination. DFA III supplementation may represent a promising strategy for mitigating the effects of low-level mycotoxin exposure in livestock production systems. Full article

(This article belongs to the Special Issue Occurrence, Toxicity, Metabolism, Analysis and Control of Mycotoxins)

23 pages, 13623 KiB

Open AccessArticle

Comprehensive Transcriptomic and m6A Epitranscriptomic Analysis Reveals Colchicine-Induced Kidney Toxicity via DNA Damage and Autophagy in HK2 Cells

by Kun Tian, Jiaxin Wen, Dongcheng Zhang, Jiaxuan Lin, Lixiang Weng, Lele Yang, Wei Zhao, Chutao Li and An Zhu

Toxins 2025, 17(8), 408; https://doi.org/10.3390/toxins17080408 (registering DOI) - 14 Aug 2025

Abstract

Colchicine is commonly prescribed for inflammation and gout, but its nephrotoxicity and underlying mechanisms remain incompletely understood. The objective of this research was to clarify the association between m6A methylation modifications and nephrotoxicity caused by colchicine. A significant decrease in HK2 cell viability was observed following colchicine treatment, and mRNA sequencing (mRNA-seq) revealed the differential expression of genes associated with DNA damage and autophagy. Further methylated RNA immunoprecipitation sequencing (MeRIP-seq) analysis revealed an association between N6-methyladenosine (m6A) modifications and the expression of genes involved in DNA damage and autophagy after colchicine exposure. Molecular docking and a molecular dynamics (MD) analysis identified ZC3H13 as a potential regulator of colchicine-induced cytotoxicity in HK2. Experimental validation confirmed that colchicine induces DNA damage and autophagy in HK2 cells, with ZC3H13 playing a significant role in these processes. In conclusion, the findings suggested that colchicine-induced damage in HK2 cells is associated with changes in m6A methylation levels in target genes and the altered expression of m6A regulator. Full article

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9 pages, 271 KiB

Open AccessArticle

Electrophysiological Evidence of Contralateral Neuromuscular Effects Following Long-Term Botulinum Toxin Therapy in Hemifacial Spasm

by Tehran Aliyeva, Mehmet Fevzi Oztekin, Yasemin Eren and Zeynep Nese Oztekin

Toxins 2025, 17(8), 407; https://doi.org/10.3390/toxins17080407 (registering DOI) - 14 Aug 2025

Abstract

Hemifacial spasm (HFS) is a cranial nerve disorder characterized by involuntary contractions of muscles innervated by the facial nerve. Botulinum toxin type A (BoNT-A) is widely used for symptom control. Although local diffusion is well established, the extent and clinical relevance of BoNT-A spread to contralateral muscles remain unclear. This study aimed to investigate the contralateral neuromuscular effects of BoNT-A in patients undergoing long-term treatment with BoNT-A. This retrospective cross-sectional study included 39 patients with HFS (mean age, 58.6 ± 8.5 years). Bilateral compound muscle action potentials (CMAPs) were recorded before and four weeks after the BoNT-A injection. Single-fiber electromyography (SFEMG) jitter and mean consecutive difference (MCD) were evaluated contralaterally using concentric needle electrodes. Patients were categorized as first-time (n = 10) or long-term (n = 29; treatment duration: 1–20 years) BoNT-A recipients. Contralateral CMAP amplitudes decreased by 21.1% post-injection (p < 0.001). MCD increased from 33.2 ± 5.6 to 37.0 ± 5.3 µs (p < 0.001), and jitter rose by 81%, from 7.9 ± 6.2 to 14.3 ± 8.1 µs (p < 0.001). The percentage increase in MCD was significantly higher in long-term versus first-time patients (12.7% vs. 7.5%; p = 0.039), suggesting a cumulative neuromuscular effect. Spontaneous myokymia or fasciculations were clinically observed in four long-term patients. These findings provide electrophysiological evidence that unilateral BoNT-A injections may induce neuromuscular transmission abnormalities in the contralateral facial muscles. This effect appears more pronounced in chronically treated individuals, highlighting the need for awareness of potential bilateral spread when planning long-term therapy. Full article

(This article belongs to the Special Issue Advances in the Treatment of Movement Disorders with Botulinum Toxins)

16 pages, 1703 KiB

Open AccessArticle

Analysis of the Proteome and Biochemistry of Venom from Tityus confluens, a Scorpion That Can Be Involved in Severe Envenomation Cases in Brazil

by Laís Corrêa Lima, Henrique Ranieri Covali-Pontes, Ohanna Gabriely Souza Leite, Renata Trentin Perdomo, Luiz Filipe Ramalho Nunes de Moraes, Ludovico Migliolo, Mauricio Nogueira Moyses, Natália Gabrielly Pereira dos Santos, Daniel Carvalho Pimenta, Mariana Soares Rodrigues, Karen Morais-Zani, Guilherme Rabelo Coelho and Malson Neilson Lucena

Toxins 2025, 17(8), 406; https://doi.org/10.3390/toxins17080406 (registering DOI) - 14 Aug 2025

Abstract

In Brazil, the annual scorpion sting cases surpass those of other neglected tropical diseases, highlighting a significant public health issue. The severity of scorpion envenomation relates to the venom’s rapid action, complex composition, species identification challenges, and limited antivenom availability. This work aimed to characterize the venom of Tityus confluens through proteomic, enzymatic, and biological analyses while also assessing its reactivity to anti-scorpion antivenom. The electrophoretic analysis revealed seven protein bands, with the most prominent bands at 30, 15, and 10 kDa. The C18-RP-HPLC analysis isolated sixteen primary fractions. The proteomic analysis identified various toxins, including potassium channel toxins, sodium channel toxins, and antimicrobial peptides, as well as other proteins such as hypotensin and metalloproteinases. Antigenic components were identified in the T. confluens venom, which displayed dose-dependent but time-independent amylolytic activity. The ATPase activity significantly increased with 1–10 μg of venom. No cytotoxic effects were observed on carcinoma or non-tumoral cell lines. The T. confluens venom features a complex protein composition rich in toxins that target ion channels and enzymes. It exhibits active enzymatic and antigenic properties, and displays low cytotoxicity. This is the first proteomic research on the composition of T. confluens venom and may provide valuable insights into understanding the clinical manifestations of scorpion stings. Full article

(This article belongs to the Special Issue Transcriptomic and Proteomic Study on Animal Venom: Looking Forward)

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19 pages, 1559 KiB

Open AccessArticle

Functional and Proteomic Characterization of Acanthophis antarcticus Venom: Evidence of Fibrinogenolytic and Serine Peptidase Inhibitory Activities

by Monica V. Falla, Enzo P. Sousa, Karen de Morais-Zani, Rodrigo Valladão, Natalia G. Santos, Nathalia C. Galizio, Mariana S. Rodrigues, Heloisa F. Almeida, Adriana R. Lopes, Mauricio N. Moises, Ivo Lebrun, Patrick J. Spancer, Daniel C. Pimenta and Guilherme R. Coelho

Toxins 2025, 17(8), 405; https://doi.org/10.3390/toxins17080405 - 13 Aug 2025

Abstract

Acanthophis antarcticus, commonly known as the death adder, is a venomous Australian snake and a member of the Elapidae family. Due to its robust body and triangular head, it was historically misclassified as a viper. Its venom is known for neurotoxic, hemorrhagic, and hemolytic effects but displays low anticoagulant activity. Although key toxins such as three-finger toxins (3FTxs) and phospholipase A₂ (PLA₂) have been previously described, no study has integrated proteomic and functional analyses to date. In this study, we conducted a comprehensive characterization of A. antarcticus venom. Reverse-phase high-performance liquid chromatography (RP-HPLC) followed by LC-MS/MS enabled the identification of nine toxin families, with 3FTxs and PLA₂ as the most abundant. Less abundant but functionally relevant toxins included Kunitz-type inhibitors, CRISP, SVMP, LAAO, NGF, natriuretic peptides, and nucleotidases, the latter being reported here for the first time based on proteomic evidence. Hydrophilic interaction chromatography (HILIC) coupled with MALDI-TOF was used to analyze polar, non-retained venom components, revealing the presence of low-molecular-weight peptides (2–4 kDa). Functional assays confirmed the enzymatic activity of HYAL, PLA₂, and LAAO and, for the first time, demonstrated inhibitory activity on serine peptidases and fibrinogenolytic activity in the venom of this species. These findings expand our understanding of the biochemical and functional diversity of this venom. Full article

(This article belongs to the Special Issue Transcriptomic and Proteomic Study on Animal Venom: Looking Forward)

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18 pages, 1147 KiB

Open AccessArticle

Geographic Variation in Venom Proteome and Toxicity Profiles of Chinese Naja atra: Implications for Antivenom Optimization

by Jianqi Zhao, Xiao Shi, Guangyao Liu, Yang Yang and Chunhong Huang

Toxins 2025, 17(8), 404; https://doi.org/10.3390/toxins17080404 - 12 Aug 2025

Abstract

Differences in venom within snake species can affect the efficacy of antivenom, but how this variation manifests across broad geographical scales remains poorly understood. Naja atra envenoming causes severe morbidity in China, yet whether intraspecific venom variation exists across mainland regions is unknown. We collected venom samples from seven biogeographical regions (spanning > 2000 km latitude). Venom lethality, systemic toxicity (organ damage biomarkers and coagulopathy), and histopathology of major organs were assessed. Neutralization by antivenom and label-free quantitative proteomics (LC-MS/MS) were also performed. The results revealed a non-uniform LD₅₀, with venom from Yunnan exhibiting the highest lethality (2.1-fold higher than venom from Zhejiang, p < 0.001). Commercial antivenom showed lower neutralization efficacy against the venom from the Yunnan, Guangxi, and Guangdong regions. Regarding organ damage and coagulopathy, venom from Yunnan caused severe liver damage, while venom from the Zhejiang region induced significant coagulopathy. Finally, proteomic profiles identified 175 proteins: venom from Yunnan was dominated by phospholipases, contrasting with eastern regions (Anhui/Zhejiang: cytotoxins CTXs > 30%). Venom from Guangdong contained higher levels of the weak neurotoxin NNAM2 (5.2%). Collectively, significant geographical divergence exists in Chinese Cobra venom composition, systemic toxicity, and antivenom susceptibility, driven by differential expression of key toxins. Our study provides a molecular basis for precision management of snakebites, and we call for optimized antivenom production tailored to regional variations. Full article

(This article belongs to the Special Issue Animal Venoms: Unraveling the Molecular Complexity (2nd Edition))

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16 pages, 574 KiB

Open AccessArticle

In Vitro Evaluation of Aflatoxin B1 Detoxification by Lactobacillus, Pediococcus, and Bacillus Strains

by Sarra Rafai, Ana Moreno, Alessandra Cimbalo, Pilar Vila-Donat, Lara Manyes and Giuseppe Meca

Toxins 2025, 17(8), 403; https://doi.org/10.3390/toxins17080403 - 11 Aug 2025

Abstract

Biologically based detoxification strategies are increasingly being explored as alternatives to conventional methods for the removal of toxic contaminants in food products. Among these, aflatoxin B1 (AFB1) is one of the most potent mycotoxins due to its high toxicity, genotoxicity, and persistence in the human body once ingested. In this study, the detoxification potential of bacterial strains belonging to the genera Lactobacillus/Pediococcus (n = 10) and Bacillus (n = 10) was evaluated using extracts from naturally contaminated corn flour. Detoxification was assessed after incubation for 12, 24, and 48 h in specific culture media. AFB1 quantification and metabolite profiling were performed at each time point using Quadrupole Time-of-Flight Mass Spectrometry (LC-QTOF-MS). The highest detoxification rates were observed with Lactobacillus curvatus 14 (L. curvatus 14) (41.1 ± 19.3%) and Pediococcus pentosaceus 4 (P. pentosaceus 4) (25.4 ± 11.3%) after 48 h, and Bacillus firmus 6 (B. firmus 6) (25.1 ± 12.9%) after 24 h. An in vitro digestion model was also applied to assess detoxification under gastrointestinal conditions. Results showed substantial AFB1 reduction at the colonic stage, reaching 72.26 ± 7.54% for P. pentosaceus 4 and 69.67 ± 9.70% for L. curvatus 14. These findings underscore the potential application of Lactobacillus, Pediococcus, and Bacillus strains in biological detoxification strategies to reduce dietary exposure to AFB1. Full article

(This article belongs to the Special Issue Aflatoxins: Contamination, Analysis and Control)

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21 pages, 3230 KiB

Open AccessArticle

Phytochemistry, Mode of Action Predictions, and Synergistic Potential of Hypenia irregularis Essential Oil Mixtures for Controlling Aedes aegypti

by Luis O. Viteri Jumbo, Wellington S. Moura, Richard D. Possel, Osmany M. Herrera, Rodrigo R. Fidelis, Bruno S. Andrade, Guy Smagghe, Gil R. Santos, Eugênio E. Oliveira and Raimundo W. S. Aguiar

Toxins 2025, 17(8), 402; https://doi.org/10.3390/toxins17080402 - 11 Aug 2025

Abstract

Aedes aegypti, also known as the yellow fever mosquito, presents a major public health challenge, highlighting the need for effective biorational agents for mosquito control. Here, we investigated the synergistic effects of essential oil mixtures derived from Hypenia irregularis that is a mint-family shrub native to Brazil’s Cerrado biome, known as “alecrim do Cerrado”, in combination with essential oils from noni (Morinda citrifolia), Brazilian mint (“salva-do-Marajó”, Hyptis crenata), and lemongrass (Cymbopogon citratus) against Ae. aegypti. We conducted phytochemical analyses and assessed larvicidal, repellent, and oviposition deterrent activities. Using in silico methods, we predicted molecular interactions between key essential oil components and physiological targets involved in repellent action (odorant-binding protein AeagOBP1 and olfactory receptor Or31) and larvicidal activity (GABA and octopamine receptors, TRP channels, and acetylcholinesterase [AChE]). Major compounds identified included octanoic acid (23%; Hipe. irregularis × M. citrifolia), 2,5-dimethoxy-p-cymene (21.9%; Hipe. irregularis × Hypt. crenata), and citral (23.0%; Hipe. irregularis × C. citratus). Although individual oils showed strong larvicidal activity (Hipe. irregularis LC₅₀ = 2.35 µL/mL; Hypt. crenata = 2.37 µL/mL; M. citrifolia and C. citratus = 2.71 µL/mL), their mixtures did not display synergistic effects. Similarly, repellency and oviposition deterrence were comparable to DEET for individual oils but were not enhanced in mixtures. Notably, the Hipe. irregularis × C. citratus essential oil blend reduced oviposition deterrence. Molecular docking confirmed strong binding of major oil components to AeagOBP1 and Or31, supporting their role in repellency. For larvicidal effects, AChE showed the highest predicted binding affinity. Overall, our findings suggest that H. irregularis, Hypt. crenata, C. citratus, and M. citrifolia (alone or in 1:1 mixture) are promising, sustainable agents for A. aegypti control. Full article

(This article belongs to the Section Plant Toxins)

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20 pages, 345 KiB

Open AccessReview

Botulinum Toxin for the Treatment of Tremors

by Steven Bellows and Joseph Jankovic

Toxins 2025, 17(8), 401; https://doi.org/10.3390/toxins17080401 - 11 Aug 2025

Abstract

Tremor, an oscillatory movement disorder, is commonly encountered in clinical practice in the setting of a variety of etiologies, such as essential tremor and Parkinson’s disease. Despite its high prevalence, treatment options are somewhat limited. Oral medications are often ineffective or limited by side effects, and other treatments, such as deep brain stimulation, are more invasive and costly. Botulinum toxin (BoNT) injections are a well-established therapy in the treatment of dystonia, but its use in the treatment of tremors has not been fully explored. In this review, we discuss the available randomized controlled trials and open-label evidence for the use of BoNT in various tremor etiologies, as well as its injection techniques. While essential tremor is the most studied condition, other tremor etiologies and tremor types such as Parkinson’s disease, head tremor, voice tremor, proximal tremor, and tremor due to dystonia and multiple sclerosis have been studied as well. Botulinum toxin injections have provided evidence of significant benefit in outcomes in several trials among these indications, but transient weakness remains a common adverse effect. There is a paucity of well-designed trials as many published studies have relatively small cohorts and results are additionally limited by heterogenous outcome measures, dosages, muscle selection techniques and methods of injection. Full article

(This article belongs to the Special Issue Biological and Pharmacological Properties of Botulinum Neurotoxins)

14 pages, 3876 KiB

Open AccessArticle

Ricin Toxicity to Intestinal Cells Leads to Multiple Cell Death Pathways Mediated by Oxidative Stress

by Francesco Biscotti, Massimo Bortolotti, Federica Falà, Antimo Di Maro, Andrea Bolognesi and Letizia Polito

Toxins 2025, 17(8), 400; https://doi.org/10.3390/toxins17080400 - 9 Aug 2025

Abstract

Ricin, a type 2 ribosome-inactivating protein, is a lethal toxin found in castor bean seeds. Although the systemic toxicity of ricin has been extensively studied, its localized effect on the gastrointestinal tract remains a critical concern, particularly in the case of oral ingestion. This study investigates the cytotoxic effects of ricin on human intestinal epithelial cell lines and its impact on epithelial barrier integrity. Ricin cytotoxicity was assessed on the intestinal-derived HT29 and Caco-2 cell lines using dose– and time–response assays, while the epithelial integrity was evaluated via Trans-Epithelial Electrical Resistance (TEER) measurements in Caco-2 monolayers. Cell death was determined through flow cytometry analysis, and the protective effects of cell death inhibitors and antioxidant scavengers were investigated on ricin-intoxicated cells. Ricin showed high cytotoxicity on HT29 and Caco-2 cells, with EC₅₀ values in the nM range after 24–72 h of intoxication. Moreover, ricin strongly reduced TEER values in Caco-2 cells at 0.1–1 nM after 24 h of treatment. At a 1 nM concentration, ricin cytotoxicity can be significantly prevented by pre-incubating cells with the cell death inhibitors Z-VAD or necrostatin-1 and the antioxidant scavenger catalase, butylated hydroxyanisole or sodium pyruvate, demonstrating the involvement of apoptosis/necroptosis and oxidative stress in ricin cell death pathways and mechanisms. Full article

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

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12 pages, 399 KiB

Open AccessArticle

Aflatoxin M₁ Levels in Milk and Urine Samples of Nursing Mothers in Bangladesh: Exposure Assessment of Infants

by Humaira Rashid Tuba, Sohel Rana, Khandaker Atkia Fariha, Gisela H. Degen and Nurshad Ali

Toxins 2025, 17(8), 399; https://doi.org/10.3390/toxins17080399 - 8 Aug 2025

Abstract

Breast milk is the ideal source of nutrition for infant growth and development. However, when nursing mothers consume aflatoxin B₁ (AFB₁)-contaminated food, the hydroxylated form aflatoxin M₁ (AFM₁) is transferred to breast milk and urine. AFB₁ and its metabolite AFM₁ are potent carcinogens and can pose significant risks to food safety and public health worldwide. This study determined the prevalence of AFM₁ in the breast milk and urine of nursing mothers in Bangladesh and estimated infant exposure to this toxin. Breast milk and urine samples (72 each), collected from nursing mothers in three different regions of the country, were analyzed for AFM₁ occurrence via a sensitive enzyme-linked immunosorbent assay (ELISA). AFM₁ was present in 88.9% of urine samples, with a mean concentration of 109.9 ± 52.8 pg/mL, ranging from 40.0 to 223.8 pg/mL. AFM₁ was also detected in 50% of the breast milk samples, with a mean concentration of 4.6 ± 0.7 pg/mL, ranging from 4.0 to 6.1 pg/mL. A strong correlation (r = 0.72) was observed between milk and urinary AFM₁ levels, indicating these as suitable biomarkers of AFB₁ exposure. Yet, no significant correlations were identified between AFM₁ levels in either milk or urine and the food items typically consumed by nursing mothers. The average estimated daily intake (EDI) for AFM₁ with breast milk was 0.59 ng/kg bw/day, with no significant difference between infants (0.57 ng/kg bw/day) and toddlers (0.65 ng/kg bw/day). A comparison of computed EDI ranges for AFM₁ with a proposed tolerable daily intake value resulted in Hazard Indices below 1 for all exposure scenarios. This indicates that the AFM₁ concentrations in breast milk from three regions of Bangladesh raise no concern. Also, the measured levels were far lower than the maximal levels set in the EU regulations for AFM₁ in dairy milk and infant formula (50 ng/kg and 25 ng/kg, respectively). Full article

(This article belongs to the Special Issue Occurrence, Toxicity, Metabolism, Analysis and Control of Mycotoxins)

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15 pages, 2398 KiB

Open AccessArticle

Heating up the Blunts: Prothrombin Activation, with Factor Va as an Obligate Cofactor, Is the Dominant Procoagulant Mechanism of Blunt-Nosed Viper Venoms (Macrovipera Species)

by Patrick S. Champagne, Lorenzo Seneci and Bryan G. Fry

Toxins 2025, 17(8), 398; https://doi.org/10.3390/toxins17080398 - 8 Aug 2025

Abstract

Venoms of the Palearctic vipers in the Macrovipera genus cause severe procoagulant clinical effects, yet the precise molecular targets remain incompletely defined. To fill this toxicological knowledge gap, we tested five Macrovipera venoms—M. lebetina cernovi, M. l. obtusa, M. l. turanica (Turkmenistan and Uzbekistan localities), and M. schweizeri—using plasma clotting assays, Factors VII, X, XI, and XII and prothrombin zymogen activation assays, and SDS-PAGE to visualise Factor V (FV) cleavage. All venoms induced extremely rapid clot formation (10.5–12.5 s) compared with the negative control (spontaneous clotting) of 334.6 ± 3.6 s) and the positive control (kaolin trigger) of 55.8 ± 1.9 s. Activation of FVII or FXI was negligible, whereas consistent FX activation and species-variable FXII activation, both moderate, were observed. Prothrombin remained inert in the absence of cofactors, but the presence of FV or FVa elicited potent thrombin generation. SDS-PAGE confirmed proteolytic conversion of the 330 kDa FV zymogen into the ~105 kDa heavy and ~80 kDa light chains of FVa by the venoms of all species. This data demonstrates that Macrovipera venoms rely on a dual enzyme strategy: (i) activation of FV to FVa by serine proteases and (ii) FVa-dependent prothrombin activation by metalloproteases. These results reveal that prothrombin activation is the dominant procoagulant pathway and overshadows the historically emphasised FX activation. This mechanism mirrors, yet is evolutionarily independent from, the FXa:FVa prothrombinase formation seen in Australian elapid venoms, highlighting convergent evolution of cofactor-hijacking strategies among snakes. The discovery of potent FVa-mediated prothrombin activation in Macrovipera challenges existing paradigms of viperid venom action, prompts re-evaluation of related genera (e.g., Daboia), and underpins the design of targeted antivenom and therapeutic interventions. Full article

(This article belongs to the Special Issue Toxins from Venoms and Poisons)

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16 pages, 1518 KiB

Open AccessArticle

T-2 Toxin-Induced Hepatotoxicity in HepG2 Cells Involves the Inflammatory and Nrf2/HO-1 Pathways

by Mercedes Taroncher, Felipe Franco-Campos, Yelko Rodríguez-Carrasco and María-José Ruiz

Toxins 2025, 17(8), 397; https://doi.org/10.3390/toxins17080397 - 8 Aug 2025

Abstract

The T-2 toxin is one of the most toxic mycotoxins, to which the population is exposed through the diet. T-2 toxins are especially found in cereals and cereal-based products. To deepen our understanding of the mechanisms of T-2 toxin action, the morphological changes, oxidative stress, and inflammatory response of this mycotoxin have been evaluated in HepG2 cells. The mRNA and protein expression levels of inflammatory cytokines such as IL-1β, IL-6, and TNF-α and proteins such as Nrf2 and HO-1 were analyzed after T-2 exposure (7.5, 15, and 30 nM) by qPCR and Western blot assays. Firstly, changes in the morphology of HepG2 cells after T-2 exposure from circular to elongated shape were observed in a concentration-dependent manner by indirect immunofluorescence. These alterations may reflect early signs of cell stress. The results revealed an upregulation of the mRNA of IL-1β, IL-6, and TNF-α after T-2 exposure, with the highest increase in TNF-α after 30 nM T-2, suggesting a proinflammatory effect. Regarding the oxidative response, HO-1 at the lowest T-2 concentration was upregulated. However, the Nrf2 at all T-2 concentrations tested was downregulated. These findings were corroborated by Western blot analysis. These results confirm that T-2 hepatotoxicity produces an increase in key inflammatory cytokines, modulates the Nrf2/HO-1 pathway, and produces morphological changes in HepG2 cells. The next step would be to test whether a co-exposure of natural antioxidants with T-2 exerts a cytoprotective effect. Full article

(This article belongs to the Special Issue Advances in Mycotoxin Determination: From Risk Assessment to Global Management Strategies)

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14 pages, 510 KiB

Open AccessArticle

Is There a “Non-Motor Effect” of Botulinum Toxin Treatment in Cervical Dystonia in Addition to Its Effects on Motor Symptoms?

by Małgorzata Dudzic, Anna Pieczyńska, Artur Drużdż, Anna Rajewska and Katarzyna Hojan

Toxins 2025, 17(8), 396; https://doi.org/10.3390/toxins17080396 - 6 Aug 2025

Abstract

The efficacy of botulinum toxin A (BoNT) in alleviating motor symptoms of cervical dystonia (CD) has been well established, and it is the treatment of choice in this disease. Lately, the effect of BoNT on non-motor symptoms (NMS) such as cognitive function, depression, anxiety, pain, and sleep disturbance has been observed in patients with CD. A comprehensive clinical and functional assessment of motor (dystonia severity, gait) and non-motor symptoms (cognitive functions, depression, anxiety, sleep, and pain) has been performed in a total of 34 adult patients with cervical dystonia before and after BoNT treatment. Results have also been compared to a control group. Significant improvements in the scales assessing dystonia severity have been observed, which is in line with previous studies on the effect of BoNT on motor symptoms in dystonia. Interestingly, the results also clearly indicate that BoNT has a positive effect on NMS. Among the studied non-motor domains, depression and cognitive functions improved the most after the treatment procedure. The study highlights the potential of BoNT to positively influence non-motor symptoms in patients with cervical dystonia, although its effect on various NMS is not equal. Full article

(This article belongs to the Special Issue Advances in the Treatment of Movement Disorders with Botulinum Toxins)

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20 pages, 2960 KiB

Open AccessArticle

Effectiveness of Kaolinite with and Without Polyaluminum Chloride (PAC) in Removing Toxic Alexandrium minutum

by Cherono Sheilah Kwambai, Houda Ennaceri, Alan J. Lymbery, Damian W. Laird, Jeff Cosgrove and Navid Reza Moheimani

Toxins 2025, 17(8), 395; https://doi.org/10.3390/toxins17080395 - 6 Aug 2025

Abstract

Alexandrium spp. blooms and paralytic shellfish poisoning pose serious economic threats to coastal communities and aquaculture. This study evaluated the removal efficiency of two Alexandrium minutum strains using natural kaolinite clay (KNAC) and kaolinite with polyaluminum chloride (KPAC) at three concentrations (0.1, 0.25, and 0.3 g L⁻¹), two pH levels (7 and 8), and two cell densities (1.0 and 2.0 × 10⁷ cells L⁻¹) in seawater. PAC significantly enhanced removal, achieving up to 100% efficiency within two hours. Zeta potential analysis showed that PAC imparted positive surface charges to the clay, promoting electrostatic interactions with negatively charged algal cells and enhancing flocculation through Van der Waals attractions. In addition, the study conducted a cost estimate analysis and found that treating one hectare at 0.1 g L⁻¹ would cost approximately USD 31.75. The low KPAC application rate also suggests minimal environmental impact on benthic habitats. Full article

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

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24 pages, 3858 KiB

Open AccessReview

Emerging Strategies for Aflatoxin Resistance in Peanuts via Precision Breeding

by Archana Khadgi, Saikrisha Lekkala, Pankaj K. Verma, Naveen Puppala and Madhusudhana R. Janga

Toxins 2025, 17(8), 394; https://doi.org/10.3390/toxins17080394 - 6 Aug 2025

Abstract

Aflatoxin contamination, primarily caused by Aspergillus flavus, poses a significant threat to peanut (Arachis hypogaea L.) production, food safety, and global trade. Despite extensive efforts, breeding for durable resistance remains difficult due to the polygenic and environmentally sensitive nature of resistance. Although germplasm such as J11 have shown partial resistance, none of the identified lines demonstrated stable or comprehensive protection across diverse environments. Resistance involves physical barriers, biochemical defenses, and suppression of toxin biosynthesis. However, these traits typically exhibit modest effects and are strongly influenced by genotype–environment interactions. A paradigm shift is underway with increasing focus on host susceptibility (S) genes, native peanut genes exploited by A. flavus to facilitate colonization or toxin production. Recent studies have identified promising S gene candidates such as AhS5H1/2, which suppress salicylic acid-mediated defense, and ABR1, a negative regulator of ABA signaling. Disrupting such genes through gene editing holds potential for broad-spectrum resistance. To advance resistance breeding, an integrated pipeline is essential. This includes phenotyping diverse germplasm under stress conditions, mapping resistance loci using QTL and GWAS, and applying multi-omics platforms to identify candidate genes. Functional validation using CRISPR/Cas9, Cas12a, base editors, and prime editing allows precise gene targeting. Validated genes can be introgressed into elite lines through breeding by marker-assisted and genomic selection, accelerating the breeding of aflatoxin-resistant peanut varieties. This review highlights recent advances in peanut aflatoxin resistance research, emphasizing susceptibility gene targeting and genome editing. Integrating conventional breeding with multi-omics and precision biotechnology offers a promising path toward developing aflatoxin-free peanut cultivars. Full article

(This article belongs to the Special Issue Strategies for Mitigating Mycotoxin Contamination in Food and Feed)

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20 pages, 312 KiB

Open AccessArticle

Pimelea and Its Toxicity: A Survey of Landholder Experiences and Management Practices

by Rashid Saleem, Shane Campbell, Mary T. Fletcher, Sundaravelpandian Kalaipandian and Steve W. Adkins

Toxins 2025, 17(8), 393; https://doi.org/10.3390/toxins17080393 - 6 Aug 2025

Abstract

Pimelea is one of the highly toxic plants in Australia, particularly affecting cattle. It contains simplexin, a potent toxin that can cause Pimelea poisoning (St. George Disease) in livestock. A survey was conducted to assess the current impact of Pimelea on livestock production, pasture systems, and financial losses among agricultural producers. In addition, information was also sought about the environmental conditions that facilitate its growth and the effectiveness of existing management strategies. The survey responses were obtained from producers affected by Pimelea across nine different Local Government Areas, through three States, viz., Queensland, New South Wales, and South Australia. Pimelea was reported to significantly affect animal production, with 97% of producers surveyed acknowledging its detrimental effects. Among livestock, cattle were the most severely affected (94%), when compared to sheep (13%), goats (3%), and horses (3%). The presence of Pimelea was mostly observed in spring (65%) and winter (48%), although 29% of respondents indicated that it could be present all year-round under favorable rainfall conditions. Germination was associated with light to moderate rainfall (52%), while only 24% linked it to heavy rainfall. Pimelea simplex F. Muell. was the most frequently encountered species (71%), followed by Pimelea trichostachya Lindl. (26%). Infestations were reported to occur annually by 47% of producers, with 41% noting occurrences every 2 to 5 years. Financially, producers estimated average annual losses of AUD 67,000, with 50% reporting an average of 26 cattle deaths per year, reaching up to 105 deaths in severe years. Some producers were spending up to AUD 2100 per annum to manage Pimelea. While chemical and physical controls were commonly employed, integrating competitive pastures and alternative livestock, such as sheep and goats, was considered as a potential management strategy. This study reiterates the need for further research on sustainable pasture management practices to reduce Pimelea-related risks to livestock and agricultural production systems. Full article

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

14 pages, 504 KiB

Open AccessArticle

Comparative Efficacy of pHA130 Haemoadsorption Combined with Haemodialysis Versus Online Haemodiafiltration in Removing Protein-Bound and Middle-Molecular-Weight Uraemic Toxins: A Randomized Controlled Trial

by Shaobin Yu, Huaihong Yuan, Xiaohong Xiong, Yalin Zhu and Ping Fu

Toxins 2025, 17(8), 392; https://doi.org/10.3390/toxins17080392 - 5 Aug 2025

Abstract

Protein-bound uraemic toxins (PBUTs), such as indoxyl sulphate (IS) and p-cresyl sulphate (PCS), are poorly cleared by conventional haemodialysis (HD) or haemodiafiltration (HDF). Haemoadsorption combined with HD (HAHD) using the novel pHA130 cartridge may increase PBUT removal, and this trial aimed to compare its efficacy and safety with HDF in patients with end-stage renal disease (ESRD). In this single-centre, open-label trial, 30 maintenance HD patients were randomized (1:1:1) to HDF once every two weeks (HDF-q2w), HAHD once every two weeks (HAHD-q2w), or HAHD once weekly (HAHD-q1w) for 8 weeks, with the primary endpoint being the single-session reduction ratio (RR) of IS. The combined HAHD group (n = 20) demonstrated a significantly greater IS reduction than the HDF-q2w group (n = 10) (46.9% vs. 31.8%; p = 0.044) and superior PCS clearance (44.6% vs. 31.4%; p = 0.003). Both HAHD regimens significantly reduced predialysis IS levels at Week 8. Compared with HDF, weekly HAHD provided greater relief from pruritus and improved sleep quality, with comparable adverse events among groups. In conclusion, HAHD with the pHA130 cartridge is more effective than HDF for enhancing single-session PBUT removal and alleviating uraemic symptoms in patients with ESRD, with weekly application showing optimal symptomatic benefits. Full article

(This article belongs to the Section Uremic Toxins)

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15 pages, 1507 KiB

Open AccessArticle

Determination of Fumonisins B1 and B2 in Food Matrices: Optimisation of a Liquid Chromatographic Method with Fluorescence Detection

by Óscar Cebadero-Domínguez, Santiago Ruiz-Moyano, Alberto Martín and Elisabet Martín-Tornero

Toxins 2025, 17(8), 391; https://doi.org/10.3390/toxins17080391 - 5 Aug 2025

Abstract

Fumonisins, primarily produced by Fusarium spp. and Aspergillus section nigri, are common contaminants in maize, cereal grains, and other processed and derived products, representing a significant risk to food safety and public health. This study presents the development and optimisation of a high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for the quantification of fumonisin B1 (FB1) and B2 (FB2) in various food matrices. In contrast with conventional protocols employing potassium phosphate buffers as the mobile phase, the proposed method utilises formic acid, offering enhanced compatibility with liquid chromatography systems. An automated online precolumn derivatisation with o-phthaldialdehyde (OPA) was optimised through experimental design and response surface methodology, enabling baseline separation of FB1 and FB2 derivatives in less than 20 min. The method demonstrated high sensitivity, with limits of detection of 0.006 µg mL⁻¹ for FB1 and 0.012 µg mL⁻¹ for FB2, and excellent repeatability (intraday RSD values of 0.85% and 0.83%, respectively). Several solid-phase extraction (SPE) strategies were evaluated to enhance sample clean-up using a variety of food samples, including dried figs, raisins, dates, corn, cornmeal, wheat flour, and rice. FumoniStar Inmunoaffinity columns were the only clean-up method that provided optimal recoveries (70–120%) across all tested food matrices. However, the MultiSep™ 211 column yielded good recoveries for both fumonisins in dried figs and raisins. Additionally, the C18 cartridge achieved acceptable recoveries for both fumonisins in dried figs and wheat flour. Full article

(This article belongs to the Section Mycotoxins)

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