Toxins doi: 10.3390/toxins18090393

Authors: Bazbek Davletov Alexandr Ignatchenko Aisha Zhantleuova Rashid A. Giniatullin

Alpha-latrotoxin (α-LTX) is the principal vertebrate-specific neurotoxin in widow spider (Latrodectus) venom and a powerful presynaptic secretagogue. Although its receptors, pore-forming activity and stimulation of neurotransmitter release have been studied extensively in central and motor neurons, its relationship to the severe, persistent pain of latrodectism remains poorly understood. This focused review re-examines α-LTX from a nociceptive perspective. The available evidence supports a model in which α-LTX binds adhesion G protein-coupled receptor L1 (ADGRL1; latrophilin-1) and neurexin-1α on susceptible sensory neurons; inserts a large cation-permeable pore; and promotes membrane depolarisation, calcium entry and release of pain-associated neuropeptides. The transcript-level gene expression of α-LTX receptor genes in dorsal root ganglion neurons and evidence of toxin-evoked neuropeptide release provide a molecular basis for direct nociceptor activation, although functional validation in defined nociceptor subclasses remains necessary. A comparison with the nociceptive ion channels transient receptor potential vanilloid 1 (TRPV1) and transient receptor potential ankyrin 1 (TRPA1) highlights their convergence on calcium-dependent sensory excitation, while distinguishing α-LTX from toxins that modulate endogenous channels. We propose that the pain of latrodectism is a composite state in which direct nociceptor activation complements muscle spasm and tissue-derived signalling, positioning α-LTX as a distinctive probe of pain pathways.

Toxins doi: 10.3390/toxins18090392

Authors: Padmini Vaka Laharika Kappari Gokul V. Selvaraj Ramesh K. Selvaraj Todd J. Applegate Revathi Shanmugasundaram

Mycotoxins are widespread contaminants in animal feeds and continue to pose serious threats to animal welfare, production efficiency, food security, and sustainable economic growth worldwide. Chickens are highly susceptible to multiple mycotoxins, such as aflatoxins (AFLs), deoxynivalenol (DON), fumonisins (FBs), zearalenone (ZEA), ochratoxin A (OTA), and T-2 toxin. Exposure to these toxins can damage intestinal integrity, compromise immune functions, impair nutrient absorption, and ultimately reduce production efficiency even at subclinical concentrations. Conventional mycotoxin detection methods, such as enzyme-linked immunosorbent assays (ELISA) and chromatographic techniques, provide accurate quantification but remain expensive, labor-intensive, time-consuming, and inefficient for large-scale screening, particularly when masked mycotoxins are present. The frequent co-occurrence of multiple mycotoxins further complicates risk assessment and effective management. Emerging analytical technologies, including hyperspectral imaging, biosensors, Internet of Things (IoT)-based platforms, and machine-learning algorithms, offer promising advancements for rapid detection and predictive risk forecasting. This review summarizes the toxicological impacts of major mycotoxins on poultry, outlines critical challenges in detection and prevention, and evaluates current artificial intelligence (AI) and machine learning (ML) approaches for mycotoxin identification, prediction, and management. A systematic literature search conducted across PubMed, ScienceDirect and Google Scholar identified 176 unique studies published between 2010 and 2026. Key knowledge gaps include limited availability of high-quality datasets and source code, inconsistent model interpretability, and poor reproducibility across production environments. By integrating conventional toxicology with data-driven approaches, this review highlights how predictive modeling can strengthen mycotoxin surveillance and support proactive mitigation strategies in modern poultry production systems.

Toxins doi: 10.3390/toxins18090391

Authors: Farshad Mazda Shirazi Srikar R. Adhikari Vance G. Nielsen Geoffrey Thomas Smelski

Neurotoxic envenomation by rattlesnakes has been of great concern throughout the Americas and in Arizona. In addition to clinical weakness, patients sometimes display myokymia, a disordered fasciculation of skeletal muscle, which is mediated by Mojave toxin. As neurotoxic venoms have been associated with increased release of creatine kinase (CK) activity and concordant increases in length of stay (LOS) in the hospital, it was retrospectively determined if point-of-care ultrasound detected myokymia at the bite site was associated with increased CK activity, LOS, and healthcare cost compared to rattlesnake envenomations not associated with myokymia. The Arizona Poison and Drug Information Center patient database was queried for patients with point-of-care ultrasound exam results at or near the bite site from 2017 to 2021. Using point-of-care ultrasound to detect subclinical myokymia, thirty-seven patients that met inclusion criteria were determined to have myokymia (n = 16) and/or not display it (n = 21). Patients with myokymia had twice the CK activities, double the LOS, and two-fold the healthcare costs of patients without myokymia detected. These findings serve as a rationale for future investigations to link ultrasound detected myokymia with patient outcomes.

Toxins doi: 10.3390/toxins18090390

Authors: Oana Mîrza Denisia Pașca Lara Manyes Anamaria Cozma-Petruț Roxana Banc Simona Codruța Hegheș Lorena Filip Béla Kiss

Mycotoxins are secondary fungal metabolites that have continuously attracted research interest, both for their potential negative effects on animal and human health and for the economic losses they entail. Studies assessing exposure to mycotoxins have become increasingly widespread. The aim of this study was to provide a comprehensive description of mycotoxins and to evaluate the evolution of research on monitoring mycotoxin exposure using human urine samples, concerning analytical techniques and occurrence. Studies published in English over the last 25 years from four databases (Web of Science, PubMed, Science Direct, Scopus) were analyzed. Evaluating 73 original articles, a constant interest in the analysis of ochratoxin A in human urine samples could be observed (maximum level 148 ng/mL), a fact also justified by its demonstrated toxicity. The most commonly used analytical technique to evaluate mycotoxin presence in human urine was liquid chromatography coupled with high-performance detectors. The limitations of the review include the large period covered and the lack of mycotoxin concentrations expressed in ng biomarker/mL urine in some articles. Finally, an overview of how mycotoxin levels in human urine can be applied to mycotoxin exposure risk assessment was included.

Toxins doi: 10.3390/toxins18090389

Authors: Ren-Chieh Wu Kuang-Ting Yeh

Acute kidney injury (AKI) is a severe complication of Eastern Russell’s viper (Daboia siamensis) envenoming, but species-specific mechanisms, treatment effects, and long-term outcomes remain uncertain. This review addressed four questions: the human renal evidence specific to D. siamensis; venom and host pathways contributing to AKI; conclusions supported for antivenom timing and thrombotic microangiopathy (TMA); and quantifiable long-term kidney risk. PubMed and OpenAlex were searched from inception through 3 August 2026. Species-authenticated human evidence was separated from mixed-species or D. russelii studies and from venom-analytical and experimental evidence. Referral cohorts show AKI and dialysis burdens, but heterogeneous ascertainment, definitions, and care preclude pooled incidence. Proteomic and experimental studies support interacting coagulopathic, endothelial, hemodynamic, pigment, inflammatory, and direct renal pathways; no human study quantifies their relative contribution, and no D. siamensis study localizes specific venom antigens within renal tissue. Observational data support prompt use of indicated, regionally appropriate antivenom but do not validate a universal two-hour renal-protection threshold. TMA and long-term outcome evidence remain largely indirect. Serial renal and coagulation surveillance is warranted irrespective of whether injury is direct or indirect. Prospective, species-authenticated studies linking venom kinetics and toxin profiles to standardized renal phenotypes and follow-up are required.

Toxins doi: 10.3390/toxins18090388

Authors: Kuo-Hao Lo Kuo-Cheng Lu Yen-Hung Liu Yi-Chou Hou

Streptococcal toxic shock syndrome (STSS) is a fulminant manifestation of invasive group A Streptococcus infection characterized by superantigen-driven hyperinflammation, refractory shock, and rapidly progressive multiorgan dysfunction. Despite antimicrobial therapy, source control, intravenous immunoglobulin, and intensive organ support, some patients deteriorate rapidly, prompting interest in extracorporeal immunomodulation. This narrative review examines the biological rationale and clinical experience of hemoadsorption in STSS, supported by an illustrative case and comparison with published reports. Our patient developed vasopressor-dependent shock, acute kidney injury, thrombocytopenia, hepatic and myocardial injury, hyperlactatemia, and metabolic acidosis within 48 h. CytoSorb was integrated with continuous venovenous hemofiltration during ongoing deterioration, followed by hemodynamic and organ recovery; however, concurrent therapies preclude attribution of benefit to hemoadsorption. Published cases show a similar pattern, with extracorporeal escalation initiated during refractory shock and accumulating organ dysfunction rather than at a predefined biomarker threshold. Although experimental and observational evidence supports the biological activity of hemoadsorption, clinical evidence remains insufficient to establish a survival benefit. Current evidence does not support routine hemoadsorption for STSS. A rapidly progressive, multidimensional hyperinflammatory trajectory may instead provide a framework for future studies of patient selection and treatment timing.

Toxins doi: 10.3390/toxins18090387

Authors: Suguru Yamamoto

The number of patients with chronic kidney disease (CKD) has increased worldwide [...]

Toxins doi: 10.3390/toxins18090386

Authors: Ivonne Salazar-Rodríguez Iván López Patricia Glorio-Paulet Isabel Cristina Molina-Botero Arturo Gerardo Valdivia-Flores Carlos Gómez-Bravo

The strategy of adding mycotoxin binders (MTBs) to feed lactating cows to reduce the absorption of aflatoxins (AFs) in the gastrointestinal tract and reduce milk contamination by hydroxylated AF compounds, especially AFM1, to below the maximum contaminant level (MCL) has become widespread. However, the indiscriminate use of MTBs is very common. The objective of the study was to determine, under field conditions, the efficacy of an MTB in reducing AFM1 contamination in cow’s milk. Feed and raw milk were evaluated on-farm using lateral flow immunochromatography devices. In a 2 × 2 Latin square design, in two separate test periods, two diets were administered consecutively to each of twelve Holstein cows (multiparous, 220 ± 20 lactation days): 1: Basal diet naturally contaminated (120 AFB1 µg/d); 2: Basal diet plus an MTB (calcium carbonate and yeast fragments; 150 g/d). MTB intake effectively reduced the mean concentration, daily excretion, and percentage transfer rate of AFM1 in milk (0.724 ± 0.045 to 0.479 ± 0.055 µg/kg, p < 0.01; 15.6 ± 1.7 to 9.7 ± 1.5 µg/d, p < 0.05; and 13.0 ± 1.4 to 8.1 ± 1.3%, p < 0.05, respectively). These results highlight the need to evaluate, under field conditions, the effectiveness of MTBs as part of a mycotoxin contamination control program on dairy farms.

Toxins doi: 10.3390/toxins18090385

Authors: Francesca Bosco Chiara Mollea Gabriele Baldissone Micaela Demichela Davide Fissore

In the present work, the correlation between the respirometric activity and the aflatoxin synthesis of the hazelnut microbiota has been investigated. Microcosms containing a monolayer of hazelnuts were maintained at controlled conditions in terms of temperature and humidity. To monitor microbial growth, CO2 was evaluated along with the specific O2 uptake rate, RO2, measured by a respirometric sensor, usually applied in the evaluation of soil microbial activity. The respiratory quotient (RQ) was calculated and a linear relationship between RQ and produced aflatoxins was obtained. Depending on incubation time, four main clusters of RQ values were identified: for higher incubation time (15–40 days) RQ was lower than 2 and an aflatoxin amount of 0.31–0.35 μg/kg was observed. CO2, O2, and aflatoxin data have been used to develop an autoregressive model and excellent agreement between measurements and estimations was obtained. In most cases, an error lower than 1% on the estimated value of the mycotoxin amount revealed the accuracy of the evaluation of the RQ. The present work represents the first attempt to develop an effective method, fast and low-cost, for the in-line monitoring of mycotoxin contamination.

Toxins doi: 10.3390/toxins18090384

Authors: Zixin Wang Mingzhu Chen Jialei Liu Tao Wang Zhe Sun Xiaolu Jin

The ubiquitous presence of modified mycotoxins such as zearalenone-14-glucoside (ZEN-14G) in agricultural resources presents a critical challenge to livestock safety, specifically regarding their capacity to disrupt lipid metabolism and fermentation in the rumen. This study aimed to evaluate the dose- and time-dependent direct effects of ZEN-14G on rumen fermentation characteristics, nutrient disappearance, and mycotoxin biotransformation under controlled in vitro conditions, and to explore the associated microbial and metabolic responses. An in vitro batch fermentation system was established with three ZEN-14G doses, namely control (CON), low dosage (GL), and high dosage (GH), across three time points (6, 12, and 24 h). Basic fermentation parameters, fat disappearance rates, and ZEN-14G metabolites were quantified, while 16S rRNA gene sequencing and untargeted metabolomics were conducted specifically on 24 h endpoint samples from the CON and GH groups to investigate downstream mechanistic disruptions. The results showed that while baseline pH homeostasis remained unaffected across groups, ZEN-14G dose-dependently inhibited fat disappearance, with the GH group exhibiting a significant reduction throughout fermentation. Targeted quantification confirmed that ZEN-14G was predominantly deglucosylated to free ZEN and reduced to α/β-ZEL. Endpoint multi-omics revealed that although overall α/β-diversity was maintained at 24 h, ZEN-14G induced fine-scale strain-level replacement (12% shared ASVs), marked by the depletion of the key lipolytic bacterium Prevotella sp. R79. Untargeted metabolomics showed marked disruptions in sphingolipid metabolism and linoleic acid oxidation, where the accumulation of cytotoxic oxidized fatty acids (12,13-EpOME) and membrane turnover markers (sphingosine) positively correlated with enriched Bacillota. In summary, under macroscopic acid–base homeostasis, masked ZEN-14G directly impairs ruminal lipid metabolism, alters microbial community structure at the strain level, and perturbs cell membrane lipid turnover strictly within an in vitro system. These findings establish a key mechanistic baseline for masked mycotoxin biotransformation in the rumen, highlighting the necessity for future in vivo feeding trials to fully evaluate their systemic risk profile in ruminants.

Toxins doi: 10.3390/toxins18090383

Authors: Queenta Ngum Nji Patrick Berka Njobeh

Aquaculture has become the fastest-growing food-production sector worldwide, with increasing reliance on plant-based feed ingredients such as maize, soybean, wheat, rice, and oilseed meals. Although these ingredients reduce dependence on fishmeal, they are highly susceptible to fungal contamination and subsequent mycotoxin production. Global surveys reveal widespread contamination of aquafeeds with major regulated mycotoxins, including aflatoxins, fumonisins, deoxynivalenol, zearalenone, ochratoxin A, and T-2 toxin, alongside emerging toxins such as enniatins, beauvericins, and sterigmatocystin metabolites. Aflatoxin B1 is the most prevalent and toxic mycotoxin, with contamination prevalence often exceeding 80–100% in Africa and Asia with contamination levels surpassing 400 µg/kg in some regions of these continents. Fumonisins, deoxynivalenol, and zearalenone are also commonly detected, with fumonisins levels occasionally exceeding 7000 µg/kg. Mycotoxins induce diverse toxic effects in fish, including growth suppression, hepatotoxicity, nephrotoxicity, immunosuppression, oxidative stress, reproductive dysfunction, and mortality. Collectively, these findings highlight the urgent need for improved surveillance, feed management, and harmonized regulations addressing multi-mycotoxin exposure to safeguard aquaculture productivity, fish, and public health. This review synthesizes current knowledge on the global mycotoxin contamination patterns, analytical detection methods in fish feed and processed seafood as well as the toxicological effects of mycotoxins, with emphasis on species-specific responses and food safety implications.

Toxins doi: 10.3390/toxins18090382

Authors: Min Gan Peirong Chen Rui Zeng Fuhua Wang Yarong Zhao

Zearalenone (ZEN) is a mycotoxin produced by Fusarium species that is harmful to agricultural crops. Therefore, the prevention and control of ZEN contamination is an important concern for food safety. Lactobacillus paracasei 85 is a well-recognized probiotic with considerable potential for mycotoxin biocontrol owing to its diverse probiotic properties and antimicrobial activity. Based on its reported potential, we aimed to evaluate its ability to degrade ZEN using degradation assays under optimal conditions. Transcriptome sequencing, untargeted metabolomics, and integrated bioinformatics analyses were conducted to explore the molecular mechanisms underlying ZEN removal. Transcriptomic analysis showed that the strain activated central carbon and secondary metabolism during ZEN removal, and all seven differentially expressed genes closely related to carbohydrate metabolism were downregulated. Metabolomic analysis revealed that lipid and amino acid metabolism, in addition to carbohydrate metabolism, played important roles in ZEN removal. Correlation analysis between carbohydrate metabolism-related genes and their metabolites revealed that these genes were significantly associated with four metabolites: myo-inositol, guanosine, glycolic acid, and trehalose. As key nodes in the carbohydrate metabolism pathway, changes in the expression of these four metabolites and their interactions with genes may represent key regulatory factors responsible for the overall downregulation of carbohydrate metabolism.

Toxins doi: 10.3390/toxins18090381

Authors: Sandra Juárez-García Ana Isabel Fernández-Díez Rafael Mora-Medina Nahúm Ayala-Soldado Antonio Lora-Benitez Rosario Moyano-Salvago Ana María Molina-López

Mycotoxin contamination in food and feed remains a major public health concern because of its potential toxicological impact and its global economic implications. Aflatoxins and ochratoxin A are of particular concern due to their carcinogenic, hepatotoxic, and nephrotoxic effects, highlighting the need for effective surveillance systems to reduce dietary exposure in the population. This study analyzes 2479 mycotoxin-related notifications concerning both food and feed reported through the iRASFF system during the period 2020–2024, with the aim of characterizing temporal notification trends, identifying the most affected food products and geographical regions, and evaluating the evolution of control strategies implemented by competent authorities. The results did not show a statistically significant long-term increasing trend in the number of mycotoxin notifications, although interannual fluctuations may reflect both changes in contamination patterns and improvements in detection and reporting systems. Products originating outside the European Union accounted for 90.44% of notifications and showed a significantly higher mean number of notifications than products originating within the European Union. Turkey (24%) and the United States (14%) were the most frequently reported countries of origin at a global level. Aflatoxins were the most frequently detected mycotoxins in food (84%), followed by ochratoxin A (15%), mainly affecting nuts, cereals, fruits, vegetables and herbs and spices. These findings demonstrate that mycotoxins continue to represent a persistent challenge for European food safety. Strengthening surveillance systems, enhancing international cooperation, and adapting control strategies to emerging environmental and trade-related factors are essential to minimize consumer exposure, particularly to compounds with genotoxic and carcinogenic potential, and to improve risk management within the food supply chain.

Toxins doi: 10.3390/toxins18090380

Authors: Andras-Laszlo Nagy Francesca Caloni Regina Tose Kemper Sofia Gonzalez Lleras Heather Beigh Christian DeJean Maurice Matthew Georgios Paraschou

Tropical regions are characterized by exceptional biodiversity, including extremely diverse flora. Many plants produce chemical compounds that function as defense mechanisms, some of which are toxic to mammals, including livestock species. However, documentation of toxic plants in the Caribbean region remains limited. The present study aimed to identify toxic and potentially toxic plant species affecting livestock on the island of Saint Christopher (St. Kitts) through surveys of a representative number of farms across all nine parishes. An extensive literature review was performed in order to characterize the local flora and to identify plant species with known or suspected toxic potential. Farm visits were conducted using a structured surveillance questionnaire developed by the authors. During these visits, representative photographs of commonly encountered toxic plants were taken. The identified plant species were classified according to the chemical nature of their active ingredient, and their main clinical effects were described. Ten plant species (Abrus precatorius, Senna occidentalis, Catharanthus roseus, Crotalaria retusa, Datura stramonium, Manihot esculenta, Mimosa pudica, Nerium oleander, Nicotiana tabacum and Ricinus communis) were found to be widespread throughout the island and have been documented in the literature as toxic to livestock. By knowing the distribution of these species, high-risk areas and pastures can be identified, thereby supporting targeted prevention and more effective management of future plant-associated toxicosis outbreaks.

Toxins doi: 10.3390/toxins18090379

Authors: Giovanna Viticchi Lorenzo Falsetti Marco Bartolini Sergio Salvemini Gioacchino De Vanna Eleonora Beccacece Najdi Tanuzi Mauro Silvestrini

Sleep disturbances are highly prevalent in patients with chronic migraine (CM) and are increasingly recognized as major contributors to disease burden and headache chronification. Growing evidence suggests that dysregulation of calcitonin gene-related peptide (CGRP) signaling and central sensitization may represent common biological mechanisms linking migraine and sleep impairment. OnabotulinumtoxinA (OBT-A), an established preventive treatment for CM, inhibits the release of CGRP and other nociceptive mediators from trigeminal sensory neurons, thereby reducing peripheral and central sensitization. However, its effects on sleep quality remain incompletely characterized. The aim of this prospective real-world observational pilot study was to evaluate the impact of OBT-A treatment on sleep quality in patients with CM. Consecutive patients initiating OBT-A therapy at a tertiary headache center were enrolled and assessed at baseline (before the first OBT-A treatment cycle) and after three months, immediately before the second scheduled administration, using the Pittsburgh Sleep Quality Index (PSQI), alongside measures of migraine frequency, acute medication use, migraine-related disability, and headache impact. Fifteen patients completed follow-up evaluations. OBT-A treatment was associated with significant improvements across all clinical outcomes. Median monthly migraine days decreased from 18 to 10 (adjusted p = 0.006), while acute medication use, MIDAS scores, and HIT-6 scores were significantly reduced. Sleep quality improved markedly, with median PSQI scores decreasing from 10 to 6 (adjusted p = 0.006). Eleven of 15 patients (73.3%) achieved a ≥3-point reduction in PSQI, and the proportion of participants with pathological sleep quality (PSQI > 5) decreased from 86.7% to 53.3%. Although this categorical improvement did not reach statistical significance (exact p = 0.063), likely due to the limited sample size, it may nonetheless suggest a beneficial effect on sleep quality, although the lack of statistical significance warrants cautious interpretation. These findings provide preliminary real-world evidence that OBT-A treatment is associated with improved subjective sleep quality in patients with chronic migraine. Improvements in sleep quality occurred alongside reductions in migraine frequency, acute medication use, disability, and headache impact. Given the observational and uncontrolled design of the study, it remains unclear whether sleep improvement reflects a direct effect of OBT-A or occurs secondarily to improvements in migraine-related outcomes. Larger prospective controlled studies are needed to clarify the mechanisms underlying sleep improvement and its relationship with treatment response.

Toxins doi: 10.3390/toxins18090378

Authors: Sebastian Paus Andreas Funke Jürgen Hamacher Bernhard Haslinger Jan Heckelmann Chi Wang Ip Wolfgang H. Jost Anatol Kivi Stephan Klebe John-Ih Lee Ebba Lohmann Frederic Mack Axel Schramm Andrea Stenner Pawel Tacik Uwe Walter David T. Weise

Cervical dystonia (CD) is a hyperkinetic movement disorder characterized by involuntary muscle contractions leading to abnormal postures of the head and neck. The injection of botulinum neurotoxin type A (BoNT-A) into the muscles involved is the treatment of choice for CD. BoNT-A therapy of CD requires appropriate training, which has not been standardized in Germany. To address the key challenges in BoNT-A education, an expert panel developed a simplified step-by-step treatment guideline for beginners, adapted from the established Col-Cap-Concept (“Dystonia 3×3”). Based on expert consensus, recommendations were developed for the core clinical elements of BoNT-A treatment in CD—recognition of the movement pattern, identification of target muscles, development of an injection plan, ultrasound-guided injection, and dosing, taking into account the target audience of the guideline. As such, Dystonia 3×3 provides a user-friendly beginner’s guidebook, enhancing accessibility and standardization of CD treatment training. Further research is required to evaluate its clinical efficacy and suitability for training.

Toxins doi: 10.3390/toxins18090377

Authors: Ahmed H. El-Khatib Arnold Bahlmann Christoph Hutzler Stefan Weigel

Wheat, soy and pea protein isolates used for the production of plant-based meat alternatives were analyzed for 87 mycotoxins and 26 plant toxins by means of a validated QuEChERS-based LC-MS/MS multi-method. For wheat gluten, the results revealed the presence of Alternaria toxins, deoxynivalenol, enniatins, ergot alkaloids and ochratoxin A (OTA). For soy protein, beauvericin, enniatin B and OTA were the most frequently detected mycotoxins, while for pea protein, OTA was the most frequently detected mycotoxin. From plant toxins, the tropane alkaloids atropine and scopolamine were detected in wheat gluten and soy protein. The emerging Alternaria toxin alterperylenol was reported for the first time in wheat gluten (85% of samples). OTA was present at high levels in all three matrices (exceeding the EU maximum level in one wheat gluten sample). Ergot alkaloids were found in 95% of wheat gluten samples, but none exceeded the ML. The highest co-occurrence of different toxins was observed in wheat gluten (median: 22, max: 27 toxins/sample), followed by soy protein (median: five, max: nine toxins/sample) and pea protein (median: one, max: five toxins/sample).

Toxins doi: 10.3390/toxins18090376

Authors: Antonio Evidente

Forests are an indispensable resource for human existence in the different regions of the world. Forests are often concentrated in a few parts and vary in size. The most important and extensive green reserve on Earth is the Amazon rainforest. Unfortunately, the forests are continually suffering massive losses due to abiotic stresses and deforestation to obtain other arable lands and microbial diseases, particularly those caused by pathogenic fungi. Pathogenic fungi play an important role in inducing oak decline, which is one of the most relevant forest diseases globally. The pathogens also affect disease development and its global distribution. In this review, the phytotoxins produced by fungal pathogens of different oak species in various world regions are described in depth. In particular, the isolation, the chemistry and biology of phytotoxins synthesized by oak pathogenic fungi are reported, and their role in pathogenesis is also discussed. For some phytotoxins, the enantioselective synthesis, the structure–biological activity relationships, and their utilization in agro-industry and medical applications are also discussed. In addition, for the Diplodia species, the comparison between their secondary metabolite profile and taxonomy with those of fungal pathogens of other close forest plants is discussed. Thus, fungal diseases of numerous oak trees and the serious damages they cause to the forest heritage are described, as well as the chemical and biological properties and results of some of the structure–activity relationship (SAR) studies of the phytotoxins involved. In addition, for the Diplodia species both phylogenetically closely related and unrelated, comparison between their secondary metabolite profile with those of pathogens of other close forest plants, in combination with traditional taxonomic characteristics, has helped to better understand the pathogenic process relationships within the same fungal f the pathogenic process within the same fungal family.

Toxins doi: 10.3390/toxins18090375

Authors: Ekaterina N. Chernova Sophia Barinova

In recent decades, the problem of harmful algal blooms (HABs) in various water bodies has gained widespread recognition worldwide [...]

Toxins doi: 10.3390/toxins18090374

Authors: Xue Zhang Esa Abiso Godana Kaili Wang Hongyin Zhang Qiya Yang

Aflatoxins (AFs) are highly carcinogenic mycotoxins produced by toxigenic fungi, posing serious threats to food safety, animal production, and human health. Microbial aflatoxin control has recently emerged as a promising, sustainable, and environmentally friendly strategy for reducing aflatoxin contamination. However, growing evidence suggests that reductions in AF levels within microbial treatment systems may arise not only from direct toxin degradation, but also from inhibition of fungal growth, adsorption or sequestration processes, and suppression of AF biosynthesis. In microbial co-culture systems involving viable toxigenic fungi, exogenous microorganisms and their metabolites can establish persistent ecological stress through nutrient competition, oxidative stress, and interspecies signaling. These stresses activate fungal cell wall integrity pathways, MAPK signaling cascades, and transcriptional regulatory networks, leading to membrane remodeling, alterations in lipid and energy metabolism, and redistribution of cellular resources. As a consequence, fungal physiology progressively shifts from a growth- and toxin-production-oriented state toward a survival- and defense-oriented state, resulting in impaired growth and reduced AF biosynthesis. This review proposes the internal disintegration framework, which conceptualizes AF suppression as a progressive loss of toxin-producing capacity caused by sustained microbial-induced physiological remodeling rather than solely by fungal growth inhibition or toxin degradation. It will provide new perspectives for developing precise, efficient, and eco-friendly dual-target control strategies.

Toxins doi: 10.3390/toxins18090373

Authors: Seong-Dae Moon Dae-Sik Hwang Jung-Suk Lee Su-Ok Hwang Hans W. Paerl Baik-Ho Kim

Microcystins (MCs) are cyanobacterial hepatotoxins that contaminate bloom-affected freshwater, drinking-water sources, aquatic food webs, and recreational environments. Human evidence for chronic or repeated exposure remains fragmented across study designs and organ systems. We systematically reviewed epidemiological evidence for hepatic, renal, and metabolic outcomes and integrated supporting mechanistic toxicology. PubMed, Scopus, and Web of Science searches identified 9021 records published from 1990 to 2025; an update search was conducted in April 2026. Risk of bias was assessed using a domain-based framework adapted from the Newcastle–Ottawa Scale and OHAT. The review was not prospectively registered. The qualitative evidence architecture retained 192 studies: 34 human epidemiological studies and 158 supporting mechanistic or contextual studies. Only seven independent adjusted estimates met the one-study–one-main-estimate rule for the primary quantitative synthesis. The hepatic meta-analysis included two estimates (OR 2.13; 95% CI, 1.29–3.53), the renal finding was a descriptive single-study estimate (OR 4.81; 95% CI, 1.96–11.81), and the metabolic meta-analysis included four estimates (OR 1.73; 95% CI, 1.00–2.98) with substantial heterogeneity (I2 = 91%; prediction interval, 0.56–5.36). Mechanistic evidence supported biological plausibility but was not treated as causal human evidence. Human evidence was relatively more consistent for hepatic outcomes, although it remained limited to two independent studies with different endpoints; renal and metabolic findings remain sparse or heterogeneous.

Toxins doi: 10.3390/toxins18090372

Authors: Trang Tran-Minh Marie-France Corio-Costet Marie Laurens Nils Hocquemiller Jessica Vallance Marie-Noëlle Bonnin-Verdal Pierre Waffo-Téguo Florence Richard-Forget Vessela Atanasova

With increased mycotoxin contamination, the search for eco-friendly biocontrol strategies has become increasingly important. This study aimed to evaluate the efficiency of stilbenoid-enriched Merlot (Me) and Tannat (Ta) vine root extracts, and their major active molecule, vitisin B (VIT), against Aspergillus carbonarius growth and its ochratoxin A (OTA) production using in vitro liquid and solid, and ex vivo grape-berry assays. In liquid assay, Me and Ta (125 mg/L) and VIT (36 mg/L) significantly inhibited extracellular OTA. Additionally, VIT temporarily reduced both fungal growth and total OTA. Transcriptomics showed that VIT and Me slightly induced OTA biosynthesis genes. In solid and ex vivo assays, doses above 1 g/L decreased OTA production by up to 48% without impacting fungal growth. This lack of correlation suggested a distinct regulatory response to extract-induced stress. Importantly, the fungicide tebuconazole showed method-dependent effects. On in vitro solid medium, it significantly inhibited the fungal growth without affecting OTA yield, whereas in ex vivo berry assays, it significantly inhibited both. Our findings emphasize the potential of stilbenoid-enriched extracts as biocontrol agents targeting OTA production, highlighting the value of multi-assay evaluations. Future studies should elucidate their modes of action and optimize their efficacy.

Toxins doi: 10.3390/toxins18090371

Authors: Elodie Lassallette Philippe Guerre

Fusarium mycotoxins are major contaminants of human and animal feed and pose significant risks to health. A recent recommendation from the European Commission drastically decreased the maximum tolerated concentrations of fumonisins (FBs) and deoxynivalenol (DON) in poultry feed. The objectives of this study were to characterize the effects of a 14-day exposure to a diet containing 7.5 mg FB1 + FB2/kg, 2.5 mg DON/kg and 0.6 mg zearalenone (FDZ diet) in turkeys and chickens, and to assess the persistence of these effects following withdrawal of the contaminated diet. No clinical signs of toxicity were observed in either species, and markers of hepatic oxidative damage remained unchanged. In contrast, sphingolipidome alterations were detected, being more pronounced in turkeys than chickens and more persistent in the liver than in plasma. The C22–C24:C16 ratios measured across several sphingolipid (SL) classes emerged as sensitive biomarkers of FB exposure, whereas the sphinganine:sphingosine ratio (Sa:So) remained unchanged. Determination of the 90th percentile (P90) thresholds for SL ratios in unexposed animals provided good-to-excellent discrimination between exposed and unexposed animals. Notably, this conservative percentile-based approach detected FB exposure using the Sa:So ratio despite the absence of significant differences in group means. Multivariate analysis of the complete sphingolipidome provided the highest discriminatory power, distinguishing exposed from unexposed turkeys and chickens for up to four days after withdrawal of the FDZ diet. Finally, the biological significance of alterations in d18:1P, 18:0/2:0, 18:1/16:0, and dihydrosphingolipids induced by the FDZ diet is discussed.

Toxins doi: 10.3390/toxins18090370

Authors: Arkhom Arayawichanont Subsai Kongsaengdao

Pain is a major contributor to disability in cervical dystonia (CD) and may persist despite improvement in abnormal head posture after botulinum toxin treatment. Whether increasing the dose of Neubotulinum toxin type A (Neu-BoNT-A) provides additional analgesic benefit without compromising safety remains uncertain. In this prospective, multicenter, randomized, double-blind, two-period crossover trial, 50 adults with CD were randomized 1:1 to Neu-BoNT-A 100 U followed by 50 U or 50 U followed by 100 U, with each treatment period lasting 12 weeks. The principal between-dose TWSTRS analysis used a crossover mixed-effects model including treatment, period, and randomized sequence, with participant as a random effect. TWSTRS total, severity, and disability did not differ significantly between regimens. Neu-BoNT-A 100 U produced greater improvement in TWSTRS pain (adjusted 100 U minus 50 U difference, −1.83 points; 95% CI, −2.93 to −0.73; p = 0.0011). The pain finding remained significant in a robust GEE sensitivity analysis (difference, −1.83 points; 95% CI, −3.04 to −0.62; p = 0.003). CDIP-58 psychosocial functioning also improved more with 100 U (nominal p = 0.001). In a Holm sensitivity analysis across all 24 efficacy comparisons, TWSTRS pain and CDIP-58 psychosocial functioning remained significant (adjusted p ≈ 0.025 and 0.024, respectively); no other comparison remained significant. No serious adverse events occurred. Neu-BoNT-A 100 U did not improve overall CD severity compared with 50 U but was associated with greater pain relief and improved psychosocial functioning without compromising tolerability. These secondary findings remained significant after multiplicity adjustment but should be interpreted in the context of the nonsignificant overall TWSTRS result and require confirmation.

Toxins doi: 10.3390/toxins18090369

Authors: Vasileios G. Papatsiros

Mycotoxin contamination of feedstuffs remains one of the most persistent and economically damaging challenges confronting global animal production [...]

Toxins doi: 10.3390/toxins18090368

Authors: Mahtab Khatibi José R. Almeida Ashifa Al Juwaiser Soheil Gilabadi Ketan Patel Sakthivel Vaiyapuri

Venoms of African spitting cobras, including Naja mossambica, N. nigricincta, and N. pallida, are widely recognised for their cytotoxicity. However, their effects on platelets and blood coagulation remain poorly characterised. To address this gap, we characterised their enzymatic profiles and evaluated their effects on haemostasis and cell viability using functional blood-based and cellular assays, respectively. All three venoms markedly impaired haemostatic regulation by affecting intrinsic coagulation, significantly inhibiting platelet function, and inducing haemolysis. They also exerted potent myotoxic effects on cultured myoblasts/myotubes while causing only minimal cytotoxicity towards platelets. Fractionation by reversed-phase HPLC followed by mass spectrometry identified three-finger toxins that reproduced the potent platelet-inhibitory and myotoxic activities of the crude venoms but only partially accounted for the haemolytic and anticoagulant effects. These findings demonstrate that African spitting cobra venoms induce coordinated, multifaceted effects on human blood, with three-finger toxins serving as key functional mediators alongside other potential venom components. Overall, this study advances our understanding of the effects of spitting cobra envenoming, provides a foundation for developing improved snakebite therapies, and highlights venom-derived components as potential scaffolds for developing novel antithrombotic agents.

Toxins doi: 10.3390/toxins18090367

Authors: Gyu-Mi Jung Sung-Yong Hong Ae-Son Om

Alternariol (AOH) is a mycotoxin produced mainly by Alternaria alternata on fruits including tomatoes and strawberries as well as cereal grains including wheat and soybeans. In this study, we isolated Rhodotorula taiwanensis (R. taiwanensis) P1 from a pear and investigated the effects of incubation time and temperature on AOH reduction rates and the mechanism involved in AOH detoxification by the yeast strain. The yeast strain showed a 76.10% AOH reduction rate from the initial level (1 μg/mL) at 30 °C after 48 h of incubation. The yeast cell-free filtrate from the yeast culture did not increase the AOH reduction rate relative to the control without cell-free filtrate after 48 h. In addition, the AOH reduction rate by heat-inactivated yeast cells (74.57%) was similar to that by viable yeast cells (76.33%). The AOH detoxification test using the yeast cell wall fraction showed that AOH binds to its cell walls and that approximately 1/3 of the AOH bound onto the cell walls was extracted from them. These data strongly suggest that the AOH detoxification by the yeast strain was not due to degradation by either intracellular enzymes or extracellular enzymes of the yeast culture but was due to binding to yeast cell walls. The use of spheroplasts, in which cell walls are deficient, confirmed that AOH detoxification occurred by binding onto the cell walls of R. taiwanensis. Our data demonstrated that R. taiwanensis P1 was able to remove AOH by adsorption onto its cell walls. These results could help in the development of potential strategies to effectively mitigate AOH contamination of food.

Toxins doi: 10.3390/toxins18090366

Authors: Paola Rosalba Russo Andrea Sbarbati Emanuele Bartoletti Loredana Cavalieri Maurizio Cavallini Valentina Pinto Giovanni Salti Sheila Veronese

To date, there are no studies concerning the ear aging phases, nor adequate anti-aging protocols. The aims of this study are: (1) to propose an aesthetic classification of auricular aging; (2) to evaluate the aesthetic implications, efficacy, and safety levels of botulinum toxin application to the auricles as anti-aging treatment. A retrospective study was conducted in a single center on 40 patients’ charts to define an aesthetic ear aging classification, focusing on the muscles involved in the aging process; then an analysis on the results of the treatment of 50 healthy women (50–65 years), with the botulinum toxin type A for ear rejuvenation was conducted; the technique, visual aesthetic results, incidence of side effects, and efficiency of the treatment were presented. All procedures were performed in 2024, and subjects were monitored for the duration of the effects. Auricle aging process may be classified into 4 degrees. The treatment of 50 women with aging ears degree 1–2 permits correction of both ear protrusion and shape. In 1.5% of the auricles, mild swelling and transient bruising at the injection site were observed immediately after the treatment and resolved within 2–3 days. Patient satisfaction was high, and two external reviewers confirmed the treatment’s aesthetic efficiency. Aesthetic classification of auricle aging can be a useful tool in anti-aging decision-making. Ear treatment with botulinum toxin type A seems complementary to face treatment. The results are encouraging, but the number of subjects treated is limited. Further studies are required to outline age-related guidelines.

Toxins doi: 10.3390/toxins18090365

Authors: John T. Daugirdas

Comparisons of hemodiafiltration with hemodialysis have indicated significantly improved survival when the substitution fluid volume is greater than 23 L per treatment. This survival benefit has been ascribed to better removal of high-molecular-weight uremic toxins. However, with hemodiafiltration, predialysis levels of some high-molecular-weight toxins are only modestly reduced and remain far above levels measured in subjects with normal kidney function. In the present review, the potential role of other factors associated with hemodiafiltration that may beneficially impact survival relative to that seen with hemodialysis is explored. Factors analyzed include removal of relatively small uremic toxins such as urea, phosphate, and oxalate, a reduced incidence of intradialytic hypotension in some studies, possibly related to lower extracorporeal circuit temperature, benefits associated with use of ultrapure dialysis solution, and potential benefits associated with avoidance of thrice-weekly saline infusion for priming, volume replacement, and rinse back. The survival benefit associated with each of these factors may be small, but collectively, they may have a clinical impact.

Toxins doi: 10.3390/toxins18090364

Authors: Jiaxin Qin Qianqian Zhang Na Ren Zhenyu Su Zibin Zheng Tianzhu Yin Decheng Suo Xia Fan Liwen He Wei Zhang

Zearalenone-14-glucoside (Z14G) is a common modified mycotoxin attracting considerable attention due to its high occurrence in feed and food, and its potential to transform into the parent form zearalenone (ZEN) in vivo. However, the toxicokinetic data in ruminants remain scarce. In this study, male sheep (n = 5) were orally administered a single dose of Z14G at 2 mg·kg−1 BW. Serum, rumen fluid, urine, and fecal samples were collected at designated time points, and tissue samples were obtained at slaughter 168 h post-dosing. Concentrations of Z14G, ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), zearalanone (ZAN), and zearalenone-14-sulfate (Z14S) in these samples were determined using liquid chromatography coupled with tandem quadrupole linear ion trap mass spectrometry (LC-Qtrap-MS/MS), so as to explore the absorption, distribution, biotransformation, excretion patterns, and tissue residue profiles of Z14G. The results showed that Z14G was rapidly hydrolyzed in the rumen, with multiple metabolites detected within 1 h. Among these, β-ZOL exhibited concentrations of 154.75 ± 55.66 ng·mL−1, followed by ZEN 83.56 ± 33.36 ng·mL−1. In serum, ZEN, β-ZOL, and Z14G all reached peak concentrations at 30 min, with values of 20.25 ± 2.55, 3.01 ± 0.59 and 2.15 ± 0.78 ng·mL−1, respectively. The half-lives of all three analytes were less than 3 h, with Z14G eliminated the fastest (t1/2 = 0.94 ± 0.31 h) and ZEN the slowest (t1/2 = 2.91 ± 1.49 h). The total excretion of Z14G and its metabolites in feces and urine accounted for 0.90 ± 0.14% and 0.04 ± 0.01%, respectively. Z14G was not detected in any tissues, but ZEN was detected (1.38–3.61 ng·g−1). As the first systematic toxicokinetic investigation of Z14G in ruminants, this study provides critical data for food safety risk assessment of Z14G, and offers a theoretical basis for controlling Z14G contamination in ruminant feed.

Toxins doi: 10.3390/toxins18090363

Authors: Lorenzo Covarelli Giovanni Beccari Emilio Balducci Ida Karlsson Paula Persson Hanna Friberg Michael Sulyok Antonio Prodi Massimo Montanari Francesco Tini

Barley grain provides an ecological niche in which fungal microorganisms can develop. The presence of mycotoxigenic genera can result in the accumulation of fungal secondary metabolites. The present study aims to survey barley grain samples collected from Northern and Central Italy by assessing: (1) the presence of fungal microorganisms using high-throughput sequencing of the internal transcribed spacer region; (2) the presence and quantification, by real-time quantitative PCR (RT-qPCR), of the main Fusarium species associated with Fusarium head blight; and (3) the presence and quantification of secondary metabolites produced by Fusarium and Alternaria using liquid chromatography–tandem mass spectrometry. Both potentially pathogenic genera (mainly within the Ascomycota division) and non-pathogenic genera (within both Ascomycota and Basidiomycota divisions) were detected. Alternaria and Cryptococcus were the dominant genera within Ascomycota and Basidiomycota divisions, respectively, in both macro-areas. Some genera, such as Aureobasidium and Cryptococcus, showed interesting negative correlations with several pathogenic genera. Two important mycotoxigenic genera (Alternaria and Fusarium) were detected, with RT-qPCR that revealed the predominance of Fusarium graminearum in the Northern macro-area. Fusarium secondary metabolites exhibited generally higher accumulation in the North, whereas Alternaria secondary metabolites were more evenly distributed across Italy, reflecting the abundance of the two genera. The co-occurrence of Fusarium and Alternaria secondary metabolites within single barley grain samples was also observed.

Toxins doi: 10.3390/toxins18090362

Authors: Kana Yuasa Hidehisa Shimizu

Gut-derived indole metabolites are implicated in the gut–kidney axis, but the factors controlling the intestinal conversion of indole-3-acetic acid (IAA) to skatole remain incompletely defined. We developed a deterministic, hypothesis-generating framework that represents this conversion as a finite-pool allocation process governed by pH-dependent ecological permissiveness, terminal-conversion capacity, precursor availability, spatial progression, and competing loss. The model separates the available-pool scale from a dimensionless integrated conversion exposure, Ψ. Across 5400 loss-free scenarios spanning 10 pH profiles and graded metabolic and host-associated constraints, the distal endpoint normalized to the available pool followed the analytically derived relationship 1−exp−Ψ. Thus, distinct combinations of mechanistically relevant parameters produced the same normalized distal endpoint, demonstrating that this endpoint alone cannot uniquely identify the underlying mechanism. Competing loss further separated absolute, total-pool-normalized, and conditional outputs, showing that mechanistic interpretation depends on endpoint normalization. Analytical and numerical checks supported internal consistency. The framework was not fitted to biological data, and concentration values were used only as technical scaling references. This biologically unvalidated model generates experimentally testable hypotheses regarding the roles of pH, terminal-conversion capacity, precursor availability, and competing loss in intestinal IAA-to-skatole metabolism; it is not intended to provide physiological or clinical predictions.

Toxins doi: 10.3390/toxins18090361

Authors: Charbel Sleilaty Marilyn Hnein Teddy Lattouf Thea Gemayel Fouad Attieh Tia Kreidy Kevin Sarkis May Bark Maha Hoteit Alain Chebly Marwan Ghosn André El Khoury Jad Chémali

Aflatoxin B1 (AFB1) is a potent foodborne mycotoxin classified as a Group 1 human carcinogen by the International Agency for Research on Cancer (IARC) and is widely recognized for its causal role in hepatocellular carcinoma (HCC). Beyond its well-established hepatotoxicity, increasing evidence indicates that AFB1 also disrupts intestinal homeostasis by impairing epithelial barrier integrity, reducing mucus production, altering gut microbial composition, and disturbing microbial metabolism. These changes promote increased intestinal permeability, facilitating the translocation of bacteria and microbial products and contributing to chronic inflammation through the gut–liver axis. Recent experimental studies further suggest that alterations in short-chain fatty acid (SCFA) production and microbiota-dependent signaling pathways may actively mediate AFB1-induced intestinal and hepatic injury. Although direct evidence linking AFB1-induced dysbiosis to extrahepatic carcinogenesis remains limited, growing evidence indicates that persistent barrier dysfunction, microbial imbalance, and chronic inflammation may create a microenvironment favorable for tumor initiation and progression in tissues beyond the liver. This review critically summarizes current evidence regarding the effects of AFB1 on intestinal barrier function, gut microbiota dysbiosis, bacterial translocation, microbial metabolites, and the gut–liver axis while evaluating the mechanistic evidence supporting these interactions and highlighting the major knowledge gaps that should be addressed in future research.

Toxins doi: 10.3390/toxins18090360

Authors: Marko Bašković Vedrana Nikić Čižmek Jana Buzuk Bianka Dujić Danijela Jurić Kristina Jurković Karla Pehar Sara Vuković

Botulinum toxin, once known only as the cause of botulism, has become a versatile therapeutic agent whose use in children now extends across many medical and surgical specialties. This narrative review synthesises the clinical use of botulinum toxin in patients up to eighteen years of age. We describe its pharmacology, the non-interchangeable commercial preparations, and the weight-based dosing and safety principles specific to childhood, then examine its applications by organ system. Gastrointestinal uses include functional constipation and internal anal sphincter achalasia, persistent obstruction after surgery for Hirschsprung disease, anal fissure, and achalasia. Urological uses centre on neurogenic and idiopathic detrusor overactivity and on sphincter-directed injection for dysfunctional voiding. Neurological uses are dominated by cerebral palsy spasticity and, on considerably weaker evidence, dystonia, while head and neck uses include sialorrhoea, congenital muscular torticollis, and strabismus. We also address neonatal brachial plexus palsy together with hyperhidrosis and aesthetic use in adolescents. The evidence supporting these uses is markedly uneven. Randomised controlled trials exist for only a small number of paediatric indications, chiefly spasticity in cerebral palsy, neurogenic detrusor overactivity, and chronic sialorrhoea, whereas most remaining applications rest on observational cohorts, small case series, or extrapolation from adult practice, and we grade the certainty of evidence separately for every indication. Only a few indications are formally approved, and most remain off-label, though increasingly supported. Botulinum toxin is a reversible and generally safe adjunct, but rigorous paediatric trials are still needed.

Toxins doi: 10.3390/toxins18090359

Authors: Andrés Zamora Alexandra Rucavado Teresa Escalante José María Gutiérrez Erika Camacho

Skeletal muscle regeneration is often impaired after acute muscle damage induced by viperid snake venoms, such as that of Bothrops asper, a medically relevant species in Latin America. It has been shown that traces of venom that remain in the damaged muscle affect myogenic cells in culture, raising the possibility of inhibiting these toxins during the regenerative process to improve regeneration. Using a mouse model of myonecrosis and regeneration, we evaluated the effects of Varespladib (a phospholipase A2 inhibitor) or Marimastat (a metalloproteinase inhibitor) on muscle regeneration when administered intravenously 24 h after the onset of myonecrosis, i.e., after muscle damage has occurred. The regenerative process was evaluated 14 and 28 days after venom injection. Marimastat, or a combination of both inhibitors, increased the number and diameter of regenerating muscle fibers and reduced tissue fibrosis compared to tissues from mice receiving only venom, as judged by qualitative and quantitative histological assessment. These initial results underscore the deleterious role of traces of venom components, especially metalloproteinases, in damaged muscles during muscle regeneration.

Toxins doi: 10.3390/toxins18090358

Authors: Andrea Felice Armenti Giovanni Salti

Botulinum neurotoxin type A (BoNT-A) is used across coexisting craniocervical conditions, and therapeutic response is often overinterpreted as evidence of a shared generator. The bruxism literature shows why. Across six controlled studies with event-level outcome measurement, reported effects on event frequency track not dose or injection field but whether the rule used to detect an event could follow the amplitude the toxin had just reduced; none yet combines a placebo arm with an amplitude-independent event definition. This targeted critical narrative review interprets representative evidence mechanistically rather than assessing efficacy. It examines chronic migraine, tension-type headache, myogenous temporomandibular disorders, and bruxism, with somatosensory tinnitus as a cross-modal boundary case. Efficacy is protocol-specific in chronic migraine and uncertain elsewhere. Tracing studies locate somatosensory routes to the cochlear nucleus in the spinal trigeminal and dorsal column nuclei; a direct mesencephalic-trigeminal-to-cochlear projection has not been demonstrated in the tracing literature reviewed, so somatic–auditory plausibility does not establish the masticatory proprioceptive route invoked by muscle-targeted rationales. Because BoNT-A affects motor output, peripheral nociceptive signaling, and muscle spindle input—the last of these probably differing in availability across injection fields—we frame it as a site-dependent, multi-output perturbation. That pharmacology is established; what is offered here is the inferential framing and the anatomical constraint following from it.

Toxins doi: 10.3390/toxins18080357

Authors: Ana Juan-García Cristina Juan García

Biomonitoring is a valuable tool for assessing exposure to toxicants and understanding their potential impact on human and animal health [...]

Toxins doi: 10.3390/toxins18080356

Authors: Alessandro Picelli Rita Di Censo Nicola Smania Valentina Varalta Mirko Filippetti

Post-stroke spasticity may interfere with function and rehabilitation and contribute to pain, contracture, care burden, and disability. Botulinum toxin type A is an established focal treatment for post-stroke spasticity, but it is often introduced in the chronic phase, when persistent involuntary muscle overactivity may coexist with secondary musculoskeletal complications. This opinion paper critically examines whether early treatment with botulinum toxin should be considered a myth or an emerging clinical reality. Current evidence from meta-analyses, randomized trials, observational cohorts, and pooled analyses was reviewed with particular attention to treatment timing, treatment triggers, functional outcomes, and prevention of secondary complications. The literature supports the clinical usefulness of earlier treatment in appropriately selected patients, particularly for reducing focal involuntary muscle overactivity and resistance to passive movement, delaying symptomatic progression, slowing contracture development, and reducing pain-related complications. However, a consistent additional effect on active motor recovery has not been demonstrated, and evidence for participation and long-term functional independence remains less definitive. Moreover, “early” is heterogeneously defined across studies, ranging from the first weeks to the first year after stroke. We therefore propose moving from a rigid time-based concept of “early” treatment toward timely, target- and goal-guided intervention, integrating botulinum toxin with rehabilitation when a clinically relevant and modifiable focal neural treatment target emerges. To operationalize this concept, we propose a pragmatic set of clinical variables including the neural treatment target, its clinical impact and trajectory, passive musculoskeletal status, pain and care burden, motor recovery context, rehabilitation goals, and patient priorities.

Toxins doi: 10.3390/toxins18080355

Authors: Qingqing Zhao Zenghao Xu Xianglong Dai Xingyu Zhang Maolong Li Juan Chang Qingqiang Yin Guoyu Yang Chaoqi Liu

Deoxynivalenol (DON) and patulin (PAT) are common mycotoxins in cereals and fruits, posing health risks for animals and human beings. In order to study their liver toxicity, 24 mice were randomly assigned to four groups, with six replicates in each group (one mouse per cage). The mice were intragastrically administered with DON, PAT, DON + PAT (DP), or without DON and PAT (the control group) for 28 days, respectively. The results showed that body weight gain was significantly reduced by all toxin treatments, compared with the control group, and the lowest body weight gain was observed in the DP group. Histopathology revealed that hepatocyte damage and inflammatory infiltration were more serious in the DP group, exhibiting the highest mRNA abundances of MyD88, IFN-γ, and JAK2. The severity of hepatocyte apoptosis induced in each group followed the order: DON > DP > PAT; the severity of oxidative stress was ranked as DP > DON > PAT. Transcriptomic analysis revealed that numerous differentially expressed genes were regulated by DP treatment, which were mainly enriched in the MAPK, JAK-STAT, and transforming growth factor (TGF)-β signaling pathways. In conclusion, individual exposure to DON or PAT triggered hepatic injury, and their co-exposure further exacerbated liver damage.

Toxins doi: 10.3390/toxins18080354

Authors: Sasha Cardozo Ciaran Skerry

Peptidoglycan (PGN) is a mesh like polymer that builds and protects the bacterial cell wall. Across and even within species, PGN varies considerably in structure and composition, and this structural diversity shapes how released fragments are detected by the host. The varied structural features of PGN can also dictate its ability to act as a toxin or danger signal. Traditionally, bacterial cell wall components have been classified as toxic, based on their ability to cause extensive host cell damage, but the specific immune responses triggered by distinct PGN structural motifs when they are released into the extracellular space, and their potential direct role as toxins are not yet fully understood. Emerging evidence suggests that PGN is recognized by the host not only through its canonical di-/tripeptides, but also through novel structural motifs that are sensed by host pattern recognition receptors (PRRs). However, the mechanisms underlying the recognition of these structurally diverse fragments and their role in promoting immune activation, bacterial pathogenesis, and immune evasion remain poorly understood. In this review, we describe how structurally distinct muropeptides are synthesized, processed, and selectively sensed by host PRRs, and how differential immune recognition shapes PRR activation, thereby influencing host–pathogen interactions and infection outcomes.

Toxins doi: 10.3390/toxins18080353

Authors: Boyu Fang Miao Long

Mycotoxin contamination remains a persistent threat to food and feed safety owing to the chemical stability of many mycotoxins, frequent co-occurrence, and matrix-dependent risks. Enzymatic detoxification enables structure-targeted transformation of toxicity-determining motifs, such as epoxide rings, reactive double bonds, amide linkages, and lactone structures. However, free mycotoxin-degrading enzymes are often constrained by poor operational stability, difficult recovery, and limited adaptability to complex matrices. Metal–organic frameworks (MOFs) provide programmable microenvironments for enzyme immobilization through tunable pore structures, interfacial chemistry, and confinement effects. This review links toxic structural motifs with enzymatic transformation targets, discusses MOF–enzyme interface engineering and representative host–enzyme compatibility, and evaluates application modes including single-enzyme systems, multi-enzyme co-immobilization or cascade systems, adsorption–degradation coupling, and detection–degradation integration. Key bottlenecks involving enzyme leakage, mass-transfer limitation, real-matrix stability, scalable preparation, and biosafety are critically discussed. Rather than treating MOFs as passive enzyme carriers, this review proposes an application-oriented framework that integrates toxin structure, enzyme function, MOF interface regulation, matrix compatibility, and safety validation to guide the development of MOF-immobilized degrading enzymes for practical mycotoxin detoxification.

Toxins doi: 10.3390/toxins18080352

Authors: Raza Mohai Ud Din Xin Zhang Salwa Eman Ahmed A. Saleh Mudathir Y. Abdulrahman Hosameldeen Mohamed Husien Shahab ur Rehman Xiaodong Guo Ning Chen Mengzhi Wang

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin which can be carried over into animal products and cause deterioration of livestock productivity when fed to livestock and wildlife. This review proposes a biomarker-guided framework for improving the early assessment of AFB1 exposure and toxicological responses in animal models. Oral exposure leads to the absorption of AFB1, which is bioactivated in the liver to the reactive AFB1-exo-8,9-epoxide that causes DNA and protein adduct formation, inflammation, mitochondrial apoptosis, and other effects. Cytochrome P450 activation and glutathione-dependent detoxification are in balance in determining susceptibility species, and this balance is different for poultry, pigs, ruminants, and rodents. In addition to traditional liver enzymes and histopathology, we highlight mechanistically informative biomarkers such as metabolites of aflatoxin, DNA and albumin adduct, lipid peroxidation products, antioxidant indices, cytokines, apoptotic markers, as well as signals involved in the Nrf2/NFκB pathway. AFB1 also damages the integrity of the intestinal barrier, the maintenance of the intestinal gut microbiota, reproductive function, growth performance, and development, thus creating a gut–liver-systemic toxic cascade. Finally, an assessment of stage-targeted interventions such as aluminosilicate binders, adsorbents derived from yeast, probiotics and nano-enabled interventions is conducted as viable tools for the reduction in exposure and injury. This review offers targeted mitigation strategies for early diagnosis of aflatoxicosis in animal production systems based on a biomarker approach.

Toxins doi: 10.3390/toxins18080351

Authors: Lanfen Yang Li Li Jinwei Dao Li Yang Qi Yang

Hymenopteran venoms contain diverse proteins and peptides that shape envenomation, defense, predation, and allergic responses. Honeybee venom from Apis mellifera is well characterized, while molecular resources for the Asian giant hornet Vespa mandarinia remain less curated. We compared protein- and peptide-fraction LC-MS/MS identification datasets from V. mandarinia venom (VM-V) and A. mellifera venom (AM-V), supported by a de novo V. mandarinia transcriptome-derived database. Protein-level identification yielded 197 protein groups in VM-V PRO and 164 protein groups in AM-V PRO. AM-V contained well-recognized honeybee venom components, including phospholipase A2, hyaluronidase, venom acid phosphatase, venom dipeptidyl peptidase 4, melittin precursor, mast cell degranulating peptide precursor, secapin, and allergen Api m 6. VM-V PRO contained transcriptome-supported candidate venom-associated proteins, including venom dipeptidyl peptidase 4-like, hyaluronidase-like, venom allergen 5-like, serine protease-like, apolipophorin-like, and hexamerin-like entries. Peptide-fraction annotation was strongest in AM-V PEP, led by melittin precursor, whereas VM-V PEP remained largely unannotated. The study focuses on identification, annotation, and hypothesis-generating functional summaries, not replicate-level differential abundance. GO, KEGG, and STRING analyses organized functional annotation patterns and prioritized candidate protein groups. These data provide an identification-based comparative venomomics resource for VM-V and AM-V and a foundation for targeted validation of candidate V. mandarinia venom-associated components.

Toxins doi: 10.3390/toxins18080350

Authors: Ashley Griffin Brittany Berry Lidia Melaku Delijah Johnson Perla Guevarra Leslie A. Shack Shauna-Kay Spencer Bindu Nanduri Kedra Wallace

Pregnancy-related acute kidney injury (PR-AKI) increases the risk of chronic kidney disease (CKD) in the postpartum period, yet mechanisms driving this transition remain unclear. Uremic toxins, including indoxyl sulfate (IS), are implicated in AKI-to-CKD progression. Using a rat model of PR-AKI induced by ischemia–reperfusion on gestational day (GD) 18, we assessed IS contributions to renal injury in the postpartum. A subset of rats received the oral adsorbent AST-120 in the postpartum period to reduce IS. Additional groups received IS during pregnancy with or without AST-120 treatment in the postpartum period. Renal function, blood pressure, circulating and urinary IS concentrations, and renal histopathology were evaluated. PR-AKI resulted in sustained postpartum elevations in circulating (p = 0.03) and urinary (p < 0.03) IS, reduced urine output (p = 0.03), increased proteinuria (p < 0.0001), increased serum albumin, and increased renal fibrosis (p = 0.008) compared to normal pregnant control rats. Absorption of indole, a precursor for IS, significantly reduced urinary IS (p = 0.03), reduced serum creatinine (p = 0.006), and attenuated renal fibrosis (p = 0.002) in treated PR-AKI rats. While not significant, indole absorption improved urine output (p = 0.06) and reduced proteinuria (p = 0.07) in treated PR-AKI rats. IS administration during pregnancy recapitulated key features of postpartum CKD. Elevated IS contributes to persistent renal injury following PR-AKI. Postpartum reduction in IS with AST-120 dampens the progression of renal injury. These findings highlight uremic toxins as mechanistic drivers and potential therapeutic targets in post partum CKD following PR-AKI.

Toxins doi: 10.3390/toxins18080349

Authors: Ellen van Eerden Anna Arczewska-Wlosek Zsofia Bata Sylwester Swiatkiewicz Regiane R. Santos

The interaction between modified forms of deoxynivalenol (DON), such as DON-3-glucoside (DON-3G), and feed enzymes is underestimated. We assessed the interaction between DON and non-starch polysaccharide enzymes (NSPases) on growth performance, and excreta levels of DON and its metabolites (Experiment 1), as well as DON toxicokinetics (Experiment 2). Broiler chickens were fed maize-based diets naturally contaminated with low (LD; 516–792 μg DON/kg diet) or moderate (MD; 2395–3020 μg DON/kg diet) levels of DON. The LD and MD diets were supplemented with or without an NSPase blend. The MD diet impaired growth performance in broiler chickens up to 14 days of age, whereas no differences were observed in older birds. NSPase supplementation decreased body weight gain (BWG) in 28-day-old broiler chickens fed the MD diet and increased excreta levels of DON-3-sulfate (DON-3S). NSPase supplementation also improved the feed conversion ratio (FCR) without increasing BWG. Additionally, NSPase altered the toxicokinetic profile of DON in broiler chickens fed the MD diet, and the time to maximum plasma concentration (Tmax) of DON-3S was influenced by both NSPase supplementation and the DON level in the diet. To our knowledge, these findings provide the first evidence that NSPases influence DON toxicokinetics in poultry.

Toxins doi: 10.3390/toxins18080348

Authors: Mengyang Li Penghua Jian Jiayi Liu Yiwei Jia Hui Deng Lingxian Yi Nan Zhuang Zelin Hong Daojin Yu Bo Yang

Phytochemicals with antifungal activity against Fusarium graminearum (F. graminearum) are considered potential in-feed anti-mycotoxigenic agents that may complement conventional post-harvest control strategies. Therefore, we screened fourteen phytochemicals, including seven quinolizidine alkaloids (QAs), for anti-F. graminearum activity, evaluated their individual and combined efficacy in inhibiting mycelial growth, spore germination, and deoxynivalenol (DON) production, and explored the underlying mechanisms via transcriptomic analysis. Matrine (MT), sophoridine (SR), and sophocarpine (SC) exhibited antifungal activity against F. graminearum, with minimum inhibitory concentrations (MICs) of 5, 4, and 4 mg/mL, respectively. The three QAs at their MICs completely inhibited mycelial growth and DON production in liquid cultures, but were less effective on potato dextrose agar plates and in pig feed. Particularly pronounced antifungal activity and inhibitory effects on DON production were observed when the three QAs were applied in combination with berberine hydrochloride (BBR). The observed inhibition appears to be attributed to dysregulated expression of the TRI cluster genes and ERG biosynthesis genes, coupled with significant modulation of steroid biosynthesis (ko00100) and central carbon/lipid metabolism pathways. These findings provide preliminary experimental evidence supporting the further exploration of QA–BBR combinations to control fungal proliferation and DON accumulation in pig feed.

Toxins doi: 10.3390/toxins18080347

Authors: Ömer Faruk Yeşil Ahmet Çelik

Mycotoxins contaminate a wide range of foods and may contribute to both acute toxicity and chronic dietary exposure. Because online video increasingly shapes public interpretation of food-safety hazards, we evaluated the quality of English-language YouTube™ content on foodborne mycotoxin risks. Of 166 records screened for eligibility, 53 were excluded and 113 videos were analyzed. Two food-safety experts independently evaluated each video with an investigator-developed Video Content Quality (VCQ) checklist, the Global Quality Scale (GQS), and a five-item modified DISCERN instrument. The final sample comprised 77 company/commercial and 36 academic/noncommercial videos. After Holm adjustment across five source-group outcomes, company/commercial videos had higher VCQ and modified DISCERN scores (both adjusted p < 0.001), whereas academic/noncommercial videos had a higher interaction rate (adjusted p = 0.024). The GQS comparison did not meet the adjusted significance threshold (adjusted p = 0.066) and should not be interpreted as proof of equivalence. Video age was associated with cumulative views and likes but not with any quality score after correction. Item-level VCQ results showed that authoritative-source citation and health-effects coverage were among the least frequently awarded full-credit domains. Although the three quality measures were strongly correlated, neither interaction rate nor viewing rate was significantly associated with them. Platform engagement therefore did not serve as a reliable marker of scientific quality in this sample. Source- and region-based comparisons remain exploratory because the single-query design, broad uploader categories, and uneven source composition may have shaped the sampled videos.

Toxins doi: 10.3390/toxins18080346

Authors: José Manuel Veiga-del-Baño José Oliva Pedro Andreo-Martínez Miguel Motas Eva María Mateo José Miguel Soria María Ángeles García-Esparza

Background: Ochratoxin A (OTA) is a widespread mycotoxin with well-established nephrotoxic, immunotoxic, and carcinogenic properties. Despite decades of research, the structure, thematic evolution, and emerging directions of the scientific literature on OTA toxicity remain incompletely mapped. Methods: A bibliometric analysis was conducted using 883 publications indexed in the Web of Science Core Collection (1965–2025). Bibliometrix, VOSviewer, and BibExcel were integrated within a comparative cross-software workflow to assess publication trends, collaboration networks, thematic evolution, and thematic research directions in in vitro and in vivo OTA toxicity studies, while enabling cross-validation of the main bibliometric outputs. Results: After normalisation, authors, journals, author keywords, and Keywords Plus showed strong cross-software agreement, whereas country and institution outputs retained software dependent differences related mainly to country aggregation and affiliation parsing. Publication output increased substantially over the analysed period, particularly during the most recent decade, reflecting the growing visibility of OTA research in food safety and toxicology. The field is highly collaborative and multidisciplinary. Oxidative stress-related terms were prominent in the keyword and thematic analyses and co-occurred with terminology related to apoptosis, DNA damage, and mitochondrial dysfunction. Thematic evolution analyses showed a transition from early studies focused on nephrotoxicity and animal models toward more recent investigations addressing molecular pathways, cellular responses, and microbiota host interactions. Comparatively less prominent or emerging bibliometric themes included the gut immune axis, co-exposure to multiple mycotoxins, and the broader representation of animal-health and productivity-related research. Conclusions: The bibliometric and science mapping analyses indicate that the literature on OTA toxicity has evolved from predominantly organ, and animal model-related research, toward increasing attention to molecular, cellular, intestinal, and microbiota-related topics. These patterns describe changes in the conceptual structure of the literature rather than direct evidence of biological causality. Comparatively, less prominent themes, including human-relevant models, combined exposure scenarios, and microbiome-related research, may warrant further investigation.

Toxins doi: 10.3390/toxins18080345

Authors: Juan Blanco Ángeles Moroño Fabiola Arévalo Jorge Correa Araceli E. Rossignoli Juan Pablo Lamas

A previously overlooked isomer of okadaic acid (OA), which is structurally related to dinophysistoxin-2 (DTX2), was identified in bivalves and water columns from the Northwest Iberian Peninsula. This isomer elutes chromatographically after DTX2 and before DTX1, with mass spectrometric fragmentation patterns distinct from those of OA. It occurs at lower concentrations than OA but at levels comparable to DTX2, showing a consistent presence since monitoring by LC-MS/MS began in 2014. The abundance of this isomer correlated more strongly with OA than with DTX2, suggesting that it originates mainly from phytoplankton species that do not produce DTX2. Seasonal patterns revealed two maxima for the isomer, aligning with OA but differing from the single autumn–winter maximum of DTX2. Spatially, the isomer followed a northeast–southwest gradient along the Galician coast, similar to the OA and DTX2 distributions. The esterification rates of the isomer in mussels are lower than those of OA but similar to those of DTX2, likely resulting in slower depuration and longer persistence. Temporal trends indicate a recent increase in the concentration and seasonality of the isomer, rendering it more conspicuous. These findings highlight the growing significance of this isomer in shellfish toxin profiles throughout the studied period, underscoring the need to determine its structure and potential toxicity to better assess risks to human health.

Toxins doi: 10.3390/toxins18080344

Authors: Jin Huang Chenxi Ling Yuxuan Li Yake Wang Liyu Yang Qiuliang Xu Hongyu Deng Congcong Li

Aflatoxin 1. (AFB1) threatens animal health and dairy safety, but mammary-protective interventions remain poorly defined. We evaluated whether S-methylmethionine (SMM; commonly known as vitamin U) attenuates AFB1-associated injury using MAC-T bovine mammary epithelial cells, a mouse mammary model, biochemical assays, RT-qPCR, Western blotting, H&E staining, and exploratory ATAC-seq/RNA-seq analysis. Under 4 μg/mL (approximately 12.81 μM) AFB1 challenge, 1 mM SMM increased MAC-T cell viability by 14.1% (Tukey-adjusted p = 0.0387) and reduced LDH release by 38.2% (p = 0.000236) relative to the AFB1-only group. SMM also shifted selected redox markers toward a less injured state. Nrf2 protein abundance differed between the AF and AU groups (p = 0.049), although pathway activation and causal dependence were not tested. In mice, SMM co-treatment reduced serum ALT by 38.5% and mammary MDA by 56.4%, while increasing mammary T-SOD by 85.4% and GSH-Px by 100.9% relative to AF (p < 0.05 for the reported pairwise comparisons); inflammatory transcripts changed non-uniformly. The AU-versus-AF integration yielded four exploratory candidate genes (TNFSF10, NBEAL1, ENSBTAG00000054644, and ENSBTAG00000052086), and the selected RT-qPCR results were directionally consistent with the RNA-seq data. Thus, SMM attenuated selected AFB1-associated injury features under the tested conditions, while the underlying mechanism remains unresolved.

Toxins doi: 10.3390/toxins18080343

Authors: Md Mostafa Masud Sumyya Waliullah Abdus Sobhan Emran Ali

Aspergillus flavus is a major fungal species that contaminates a wide range of crops and produces aflatoxins under favorable environmental conditions. Several approaches have been developed for aflatoxigenic A. flavus identification and aflatoxins detection. This review summarizes the existing methods for the identification of aflatoxigenic A. flavus, along with recent progress in aflatoxins detection methods. It also highlights limitations and possible factors associated with the accurate identification of aflatoxigenic isolates. Particularly, this review underscores the utilization of advanced analytical or detection methods along with existing identification techniques to accurately identify aflatoxigenic A. flavus.

Toxins doi: 10.3390/toxins18080342

Authors: Marcello Silvestro Maria Albanese Ilaria Orologio Luigi Francesco Iannone Francesca Pistoia Stefania Battistini Gianluca Avino Martina Petracca Marina Romozzi Raffaele Ornello Diego Centonze Simona Sacco Antonio Russo on behalf of the Italian Headache Registry (RICe) Study Group on behalf of the Italian Headache Registry (RICe) Study Group

Background: A substantial proportion of patients with chronic migraine (CM) with previous failures among oral standard-of-care migraine medications do not respond to monoclonal antibodies targeting the calcitonine gene related peptide pathway (CGRP-mAbs), leaving limited evidence-based options for subsequent preventive strategies. The present multicentre real-world study evaluated the effectiveness of onabotulinumtoxin-A (BoNT-A) in CM patients with previous CGRP-mAb failure and multiple prior oral preventive failures. Methods: This prospective, observational, non-randomized multicentre study enrolled patients with CM who had failed ≥3 classes of oral preventive treatments and ≥1 CGRP-mAb due to lack of efficacy or intolerance. Participants received BoNT-A according to the PREEMPT “follow-the-pain” paradigm every three months and were followed for 6 months. Co-primary outcomes were change in monthly headache days (MHD) after three and six months and ≥50% MHD responder rates at both time points. Results: Fifty-three patients were analysed (85% female, aged 48.5 ± 15.2 years, range 20–73). Compared to the baseline, at the third month and sixth month, MHD decreased from 22.83 (SD 6.74) to 15.38 (SD 7.91; p < 0.001) and 14.55 (SD 9.35; p < 0.001), respectively. The percentages of participants achieving the response status in MMD at the third month and sixth month were 30% and 45%, respectively. In the third and sixth months, 22 patients (41.5%) and 27 patients (50.9%), respectively, converted from chronic to episodic migraine, while the prevalence of medication-overuse headache decreased from 81.1% of patients at baseline to 45.3% and 43.4%, respectively. Migraine-related impact and disability scores improved over follow-up, alongside improvements in anxiety and depressive symptoms and in migraine-specific quality of life. Conclusions: The present findings support the use of BoNT-A in difficult-to-treat patients with CM following CGRP-mAb failure and provide further evidence that its therapeutic effects may rely on mechanisms that are at least partially distinct from, and potentially complementary to, those of CGRP-targeted therapies.

Toxins doi: 10.3390/toxins18080341

Authors: Revathi Shanmugasundaram Klint W. McCafferty Laharika Kappari Xin Ye Kelsy Robinson Anthony E. Glenn

Deoxynivalenol (DON), a Fusarium mycotoxin frequently detected in poultry feeds, impairs broiler health by damaging gut integrity, altering immune response, and compromising growth performance. Insect-derived proteins, such as those from yellow mealworm (Tenebrio molitor) larvae meal (YMW), are gaining interest as an alternate protein source for broiler chickens. Hence, this study evaluated whether defatted YMW produced from larvae reared on DON-contaminated substrates can be safely included in broiler diets. A total of 400 one-day-old Cobb 700 mixed-sex broilers were assigned to four dietary treatments—(1) the control, (2) DON (15 mg/kg), (3) 2% YMW, and (4) 4% YMW—in ten replicates with 10 birds per replicate for 15 days. On day 14, performance parameters, intestinal morphology, tight-junction protein (TJP) gene expression, and CD4+ and CD8+ T-cell populations in cecal tonsils were evaluated. Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test. DON exposure reduced body weight by 1% and body weight gain (BWG) by 1.2%, decreased the CD4+: CD8+ T-cell ratio by 80%, and reduced the villus height in the jejunum by 19.8% and ileum by 16%. DON altered the TJP expression by upregulating claudin-2 mRNA expression by 2.1-fold in the jejunum and 5.2-fold in the ileum, and it downregulated claudin-4 expression by 6.4-fold in the ileum (p < 0.05). Dietary inclusion of 2% or 4% YMW from DON-contaminated substrates had no negative effect on growth performance or immune parameters and maintained intestinal morphology and TJP gene expression, comparable to the control groups. In conclusion, defatted YMW produced from larvae reared on DON-contaminated substrates can be safely used as a sustainable alternative protein ingredient in broiler diets.

Toxins doi: 10.3390/toxins18080339

Authors: Bharti Sangwan Ugochukwu Okoro Isabella Atteck Pawel Jaruga Chinwe Ekenna Michael Fasullo

VERO cells, derived from the kidney epithelium of the African green monkey, are widely used in virology, but their ability to metabolize xenobiotics is not fully understood. Since cytochrome P450 (CYP) enzymes participate in xenobiotic metabolism, we investigated which CYP genes are expressed in VERO-E6 cells. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) showed that VERO-E6 cells express CYP3A4, CYP3A5, and CYP3A7. In contrast, CYP1A1, CYP1A2, CYP1B1, CYP2E1, CYP2D6, and CYP2C9 transcripts were either not detected or at a low detection level. To determine whether the encoded enzymes have the potential to activate aflatoxin B1 (AFB1), we used artificial intelligence (AI)-based structural modeling along with molecular docking. AI modeling suggested that CYP3A enzymes can position AFB1 in an orientation compatible with the formation of the reactive intermediate, and CYP3A4 showed the most favorable predicted interaction (docking score: −16.3 kcal/mol). To demonstrate AFB1 bioactivation, we exposed VERO-E6 cells to 200 nmol/L AFB1. After 10 days, we observed about 40% cell death. Liquid chromatography–tandem mass spectroscopy (LC–MS/MS) analysis confirmed the presence of AFB1-derived DNA adducts, indicating that metabolic activation occurred in these cells. These findings support the presence of CYP-dependent AFB1 bioactivation in VERO-E6 cells. Thus, combining computational and experimental approaches elucidates xenobiotic metabolism in cells where biochemical data are limited.

Toxins doi: 10.3390/toxins18080340

Authors: Lifeng Che Bichen Miao Lijuan Suo Jie Tang

The gut microbiota and its metabolic functions are critical for intestinal homeostasis, but their integrated responses to diarrhea remain poorly understood in non-model mammals, such as the complex-toothed flying squirrel (Trogopterus xanthipes). Here, we collected fecal samples from six captive animals with diarrhea and six healthy controls; the diarrheal animals had been provided with moldy corn, but mycotoxin levels were not measured. Fecal microbiota was profiled by full-length 16S rRNA sequencing and untargeted metabolomics with integrative analyses. Diarrheal animals showed significantly decreased fecal microbial alpha-diversity and distinct community separation. LEfSe analysis identified Lawsonia intracellularis and Streptococcus macedonicus as the most enriched taxa in the diarrhea group. Untargeted metabolomics revealed 2216 differential metabolites (1931 downregulated), primarily enriched in steroid hormone biosynthesis, arachidonic acid metabolism, and bile secretion pathways. Procrustes and Mantel tests confirmed significant microbiome–metabolome concordance (M2 = 0.422, p < 0.001). Spearman correlation networks showed that L. intracellularis was positively correlated with multiple differential metabolites, while S. macedonicus exhibited predominantly negative correlations. These findings suggest that diarrhea in T. xanthipes may be associated with moldy corn exposure and is accompanied by gut dysbiosis and metabolic disturbances. L. intracellularis and S. macedonicus may be associated with disease progression through pathways linked to lipid and bile acid metabolism and inflammation. This study provides novel microbial–metabolic insights into diarrhea in a medicinal mammal, supporting disease prevention and healthy breeding.

Toxins doi: 10.3390/toxins18080338

Authors: Daniel Torrejón Angie Regalado Alex Proleón Víctor Otárola Fanny Lazo Edith Rodríguez Erasmo Colona-Vallejos Libertad Alzamora-Gonzales Jherson Cisneros-Gutierrez Jacquelyne Zarria-Romero Miryam Paola Alvarez Flores Renata Nascimento Gomes Thatiana Corrêa de Melo Ronnie G. Gavilan Javier Cárdenas Tenorio Félix A. Urra Dan E. Vivas-Ruiz Armando Yarlequé

Lys49 phospholipase A2 (Lys49-PLA2) homologues are catalytically inactive snake venom toxins with diverse biological activities, but their functional characterization requires recombinant production strategies to overcome the limitations inherent to native venom-derived proteins. Here, we produced recombinant BaMtx (rBaMtx), a Lys49-PLA2 homologue from Bothrops atrox, in the Pichia pastoris KM71 expression system and evaluated whether its biochemical and biological properties were preserved. rBaMtx was secreted into the culture medium, purified by cation-exchange chromatography, and characterized by SDS-PAGE, Western blotting, RP-HPLC (85.9% purity), and MALDI-TOF mass spectrometry, confirming a molecular mass of 13,821 Da. Despite lacking PLA2 activity, rBaMtx retained pronounced myotoxicity in vivo, inducing dose-dependent plasma creatine kinase release and skeletal muscle damage comparable to or greater than native BaMtx. In vitro, rBaMtx reduced viability of murine 4T1 breast carcinoma cells (IC50 = 60.94 µg/mL), altered cell morphology, modestly increased IL-1β release, and produced context-dependent effects in paired human tumor and non-tumor cell models, differing from native BaMtx and crude venom. These findings establish rBaMtx as a reproducible model for investigating the mechanisms and biomedical potential of catalytically inactive Lys49-PLA2 homologues.

Toxins doi: 10.3390/toxins18080337

Authors: Apurva Lad Mudit Bhatia Johnna A. Birbeck Alex Kuang Elliot Furr Judy Westrick Youngwoo Seo Dae-Wook Kang David J. Kennedy Steven T. Haller

Excessive cyanobacterial proliferation and toxin production increasingly threaten freshwater ecosystems, drinking water systems, and public health. Among cyanotoxins, microcystin-LR (MC-LR) is one of the most prevalent variants and is recognized for its hepatotoxicity, with evidence showing it can also alter gut microbiota composition. Previous studies, including our own, demonstrate that MC exposure can induce inflammation, oxidative stress, and changes in gene expression associated with immune responses, even at concentrations below guideline limits. This study investigated the protective effect of an MC-degrading bacterium, Sphingopyxis sp. IM1 (IM1) with a known enzymatic MC degradation pathway, against MC-LR-induced hepatotoxicity under in vitro conditions. Human Hep3B hepatocytes were pretreated with varying ratios of IM1 bacteria and subsequently exposed to 9.95 ppm of MC-LR for 24 h. RT-qPCR analysis demonstrated that MC-LR exposure strongly increased the expression of the inflammatory markers TNFα and TGF-β1, whereas pretreatment with IM1 significantly attenuated these responses. Mass spectrometric analysis of cell pellets and spent culture media demonstrated reduced concentrations of intact MC-LR in IM1-pretreated samples compared to MC-LR-only controls, accompanied by increased detection of tetrapeptide degradation products generated during mlr-mediated MC degradation. These results demonstrate that IM1-mediated degradation of MC-LR attenuates toxin-induced hepatotoxic and inflammatory responses, highlighting the potential of a novel microbial-based therapeutic approach to mitigate MC-induced toxicity.

Toxins doi: 10.3390/toxins18080336

Authors: Sarah C. Finch Craig Waugh D. Tim Harwood Alison R. Turnbull Andrew I. Selwood Emillie M. F. Passfield Jonathan Puddick

Nodularin is a cyanobacterial toxin predominantly produced by Nodularia spumigena Mertens. This cyanobacterium has been associated with fish, bird, and animal deaths, and the toxin can contaminate fish and shellfish intended for human consumption. With climate change, blooms of Nodularia spumigena are becoming more frequent and more widespread, meaning that the risk this cyanobacterium poses to human health must be evaluated. However, toxicity information is scarce, meaning that data from the closely related toxin class, microcystins, are currently used instead. In the current study, the LD50 of nodularin-R by intraperitoneal injection was found to be 50 µg/kg, consistent with previous reports. For the first time, the LD50 of nodularin-R by oral administration was determined using the voluntary consumption of laced cream cheese (7.5 mg/kg), which is similar to that reported for microcystin-LR (10.9 mg/kg). The use of microcystin toxicology data in food safety assessments for nodularin, therefore, seems reasonable—but it is critical that an oral repeated-dose study is performed on nodularin to better understand the risk posed by this toxin through chronic exposure.

Toxins doi: 10.3390/toxins18080335

Authors: Lee E. Voth-Gaeddert Hannah Glesener Gabriela Montenegro-Bethancourt

Decentralized agricultural systems (DAS) are essential to food security for millions of people in low-resource settings, yet most existing contaminant monitoring frameworks have been designed for centralized supply chains. This paper proposes a structured framework for integrating contaminant monitoring into DAS by (1) mapping the diverse supply chain structures and stakeholders that constitute these systems; (2) analyzing stakeholder motivations, constraints, and value propositions for monitoring adoption; and (3) identifying efficient monitoring points where these value propositions align with product quality improvement. The framework is organized around four core principles: tiered monitoring matched to supply chain position and stakeholder capacity, monitoring at stakeholder handoff points rather than within individual segments, incentive structures that make monitoring self-sustaining, and capacity building and governance as durable infrastructure. The framework is illustrated through the case of maize and aflatoxin contamination in Guatemala. It remains a conceptual framework, however, and its principles are intended to be generalizable across crops, contaminants, and geographies rather than demonstrated to be so. Empirical validation across these dimensions remains a necessary next step. The central argument is that through this framework, small, strategically placed incentives at high-leverage points in decentralized supply chains can shift system-wide quality outcomes. This can occur without requiring top-down regulatory enforcement, which is often neither feasible nor effective in these contexts.

Toxins doi: 10.3390/toxins18080334

Authors: Leah Hernandez Camillo Tancredi Strizzi Miriam Rosina Angelina Schwarz Nina Kronqvist Samsul Arefin Peter Stenvinkel Karolina Kublickiene

Chronic kidney disease is associated with neurovascular complications including cognitive impairment, potentially involving blood–brain barrier (BBB) impairment. The protein-bound uremic toxin indoxyl sulfate (IS) promotes endothelial injury, but its effects on human brain microvascular endothelial cells remain incompletely understood. Human cerebral microvascular endothelial cells (hCMEC/D3) were exposed to IS 200 and 900 μM. BBB integrity was assessed by the FITC-dextran (4 kDa) transwell permeability assay and claudin-5 immunofluorescence. Transcriptional responses were quantified by qPCR for aryl hydrocarbon (AhR) target genes, oxidative stress-associated and inflammatory markers, senescence, and junction-associated genes. Senescence-associated phenotypic changes were evaluated by SA-β-galactosidase staining and cytokine array profiling. IS increased endothelial permeability at 24 and 48 h (~1.5-fold relative to control) without evidence of cytotoxicity and reduced claudin-5 staining intensity. IS strongly upregulated AhR target genes, including CYP1A1, CYP1B1, and CYP1A2. NFE2L2 and IDO1 increased, while NFKB1 remained unchanged. SA-β-gal positivity increased, accompanied by elevated GM-CSF and G-CSF secretion, while CDKN1A decreased at IS 900 µM and CDKN2A remained unchanged. CDH5 was downregulated, TJP1 increased at 900 µM, and CLDN5 remained unchanged. These findings indicate that IS exposure is associated with impaired BBB integrity, AhR-related transcriptional responses, oxidative stress-associated transcriptional changes, junctional remodeling, and senescence-like endothelial features. However, causal attribution to individual pathways requires inhibition or knockdown studies.

Toxins doi: 10.3390/toxins18080333

Authors: Minseung Kim Sangwoon Chung John W. Christman Jiyoung Lee

Cyanotoxin events are among the most serious consequences of cyanobacterial harmful algal blooms. Microcystins (MCs), among the most prevalent cyanotoxins, are known to adversely affect human health. Recently, respiratory exposure has emerged as an important exposure route for MCs, with potential downstream effects on the gut microbiome through the lung–gut axis. In this pilot study, we investigated acute respiratory immune responses and gut microbiome alterations following MC inhalation using female C57BL/6J mice. Mice were assigned to three dose groups: control (0 µg/kg body weight), medium (25 µg/kg), and high (50 µg/kg). MC-LR was administered intranasally once daily for 3 days. Fecal samples were collected daily for 16S rRNA sequencing, and bronchoalveolar lavage (BAL) fluids were collected following sacrifice for immune cell analysis. MC exposure resulted in significantly increased monocyte counts (p < 0.1), while neutrophil, macrophage, and total cell counts did not significantly change (p > 0.1), suggesting selective lower respiratory tract inflammation. Functional prediction analysis of gut microbiota revealed significant increases (p < 0.1) in pathways associated with host health, including heme biosynthesis, sulfur oxidation, carbon metabolism, and antibiotic resistance. These findings suggest that inhaled MCs may induce respiratory inflammation and contribute to gut microbiome dysbiosis via the lung–gut axis.

Toxins doi: 10.3390/toxins18080332

Authors: Salvatore Fundarò Paola Molinari Giovanni Brunelli Maria Cazzulani Guido Dalla Costa Roberto Dell’Avanzato Roberta D’Emilio Carlo Di Gregorio Antonella Franceschelli Monica Renga Giovanni Salti Adriano Santorelli Ugo Simone Urso Ilaria Zollino

Background: Treatment of the lower face with botulinum toxin type A (BoNT-A) is particularly challenging because of the important functional roles of the involved muscles, which increases the risk of adverse events and complications. Methods: To address this issue, a panel of 14 experts conducted a three-round consensus study, including digital surveys and a formal meeting, on the aesthetic treatment of the mentalis (MT), depressor anguli oris (DAO), and platysma (PL) muscles using BoNT-A. The study was complemented by a literature review of 16 international consensus papers published between 2004 and 2024. Results: The primary indications for MT treatment that achieved strong consensus were “peau d’orange” (“golf ball”) chin appearance and hyperactivity. The Italian panel recommended a mean total dose of 8 U (distributed bilaterally), which was higher than the international mean dose of 6.5 U. The preferred injection technique consisted of a combination of intramuscular and intradermal injections. For the DAO, the strongest indication was correction of downturned oral commissures. Both the Italian panel and the international consensus reported a mean total dose of 5.6 U. Most experts (92.8%) preferred intramuscular injections, typically administered at a single injection point lateral to the marionette line in order to remain within a safe treatment zone and avoid diffusion to the depressor labii inferioris muscle. Indications for PL treatment included platysmal bands, jawline recontouring, and improvement of skin texture. The techniques discussed included treatment of both platysmal bands and the jawline. The consensus panel recommended a significantly higher mean total dose (48.4 U) than the international average (38.2 U). The paper also highlights the growing importance of ultrasound imaging as a tool for identifying individual anatomical variations in the lower facial musculature. Conclusions: As the demand for lower face rejuvenation continues to increase, standardized injection techniques and the integration of ultrasound imaging are essential for minimizing functional complications and optimizing aesthetic outcomes.

Toxins doi: 10.3390/toxins18080331

Authors: Joanna Soto de Jesus Carmen González-Keelan Peter A. Meléndez Carmen L. Cadilla Jasmine Contreras Braulio D. Jiménez-Vélez

This study assesses the impact of sublethal levels of four marine toxins ciguatoxin (CTX-1), maitotoxin-2 (MTX-2), saxitoxin (STX) and brevetoxin-2 (BTX-2) on murine hepatic detoxification enzymes expressed in mouse liver. CTX-1, BTX-2, and STX altered hepatic detoxification responses, but their effects were generally more limited or temporally variable than those observed with MTX-2. CTX-1 produced time-dependent changes in cytochromes P450 (CYPs)-associated activities, BTX-2 induced early CYP1A2 and CYP3A11 responses followed by later suppression, and STX reduced CYP1A2 and CYP3A11 while increasing microsomal reductase activities. Of these toxins, MTX-2 exhibited the highest toxicity, notably decreasing key proteins such as CYPs, including CYP1A2 and CYP3A11. CYP enzymes are vital for metabolizing important endogenous and various xenobiotic substances, including therapeutic drugs. After MTX-2 exposure, both CYP1A2 and CYP3A11 levels dropped significantly at 12 h (p < 0.0001 for CYP3A11, p < 0.0001 for CYP1A2). Histopathological analysis revealed liver damage; however, albumin mRNA levels remained stable post-MTX-2 treatment, indicating that hepatotoxicity was not the sole cause of CYP3A11 reduction. Immunohistochemical analysis displayed uniform CYP3A11 distribution across liver regions after MTX-2 treatment. This suggests that MTX-2 exposure could augment the toxicity of drugs like Aldactone, Erythromycin, and Cyclosporine that utilize this metabolic pathway in humans. This is the first type of research performed of this nature, which could add to our understanding of marine toxin metabolism. These findings provide additional toxicological insights into the effects of four marine toxins on detoxification enzymes, with particular interest in MTX-2 toxicity, and potential implications for the treatment of fish poisoning, including ciguatera fish poisoning.

Toxins doi: 10.3390/toxins18080330

Authors: Betül Aydın Münire Nazlı Höbek Başer İpek Midi Naime Evrim Karadağ Saygı Özge Keniş Coşkun

Post-stroke spasticity involves both neural and non-neural mechanisms, including structural alterations of muscles and surrounding fascia. Although botulinum toxin type A (BoNT-A) is widely used to treat focal spasticity, its effects on fascial tissue remain unclear. This retrospective observational study investigated longitudinal changes in fascial thickness after BoNT-A injection using ultrasonography. Thirty-seven patients with post-stroke spasticity were included. Fascial thickness was measured at baseline, 3 weeks, and 3 months after treatment in the injected muscle regions. Clinical outcomes were assessed using the Modified Ashworth Scale (MAS), Fugl–Meyer Assessment (FMA), and Brunnstrom stages. Repeated-measures analysis demonstrated significant reductions in fascial thickness across all evaluated regions. Pairwise comparisons showed significant decreases from baseline to week 3 and month 3, whereas continued reductions between week 3 and month 3 were observed only in the flexor digitorum superficialis fascia, the flexor digitorum superficialis–flexor digitorum profundus interfascial layer, and the deep brachialis fascia. MAS scores and upper extremity Fugl–Meyer scores improved significantly, whereas Brunnstrom stages remained unchanged. These findings suggest that the therapeutic effects of BoNT-A may extend beyond neural inhibition to include fascial remodeling. Ultrasonographic assessment of fascial thickness may represent a potential imaging biomarker for monitoring structural treatment responses in post-stroke spasticity.

Toxins doi: 10.3390/toxins18080329

Authors: Thomas Lang Adam M. Zawada Xiaoling Wang Jochen G. Raimann Manuela Stauss-Grabo Nadja Grobe

Hemodiafiltration (HDF) combines diffusion and convection to enhance solute removal compared with hemodialysis (HD). While widely adopted in Europe and parts of Asia, uptake elsewhere remains limited due to regulatory, reimbursement, and infrastructural constraints. Recent randomized trials, large observational studies, and meta-analyses have renewed interest in HDF, particularly post-dilution HDF achieving convection volumes of ≥23 L/session, in which the most consistent clinical benefits have been observed. This review summarizes the mechanistic rationale, clinical effects, and safety profile of high-volume HDF. Compared with HD, HDF provides consistently superior removal of middle molecules, whereas clearance of protein-bound uremic toxins (PBUTs) remains constrained by albumin binding and compartmental redistribution, resulting in only modest or variable advantages. Emerging strategies aimed at releasing or capturing PBUTs may address this gap. Beyond solute removal, evidence suggests that HDF may lower micro-inflammation by enhanced clearance of inflammatory mediators and reduced immune activation. When delivered with adequate infrastructure, monitoring, modern equipment, ultrapure fluids, trained staff, appropriate vascular access, and anticoagulation, high-volume HDF has a favorable safety profile and is associated with fewer intradialytic hypotension (IDH) events and improved treatment tolerability. Available evidence has not identified a consistent excess risk of device- or infection-related complications, although comparative data remain limited. Overall, current evidence supports clinically relevant advantages of high-volume HDF under optimal treatment conditions. However, important challenges remain, particularly the limited removal of PBUTs, underscoring the need for further innovation in uremic toxin management.

Toxins doi: 10.3390/toxins18080328

Authors: Alexander Gardetto Luis Thaler-Wolf

Botulinum neurotoxin type A (BoNT-A) injection for lateral canthal rhytids is among the most frequently performed nonsurgical aesthetic procedures; however, procedure-related hematoma and ecchymosis remain frequent local adverse events associated with the multiple puncture sites required in the conventional three-point technique. This prospective single-center preliminary clinical observation evaluated a single-point intramuscular injection technique designed to reduce percutaneous vascular-layer penetrations while preserving treatment territory, dose, and intramuscular target localization. A total of 152 consecutive patients (104 female and 48 male) underwent bilateral treatment between May 2025 and May 2026 using a single skin entry point per side, positioned 10–15 mm caudal to and 10 mm temporal to the lateral canthus, followed by 18–20 mm intramuscular advancement within the lateral orbicularis oculi and continuous retrograde infusion of 12 U of onabotulinumtoxinA per side. Post-procedural assessment after injection demonstrated small unilateral ecchymosis in 2 of 304 treated sides (0.66%; Clopper–Pearson 95% Confidence Interval (CI), 0.08–2.36%). After the 10-day follow-up, no persistent bruising or other adverse events were reported, and all patients confirmed satisfaction with the treatment. At the structured one-year follow-up, the satisfactory reduction in dynamic lateral canthal rhytids and continued patient satisfaction with the treatment were confirmed, while a subset elected to return for a subsequent treatment session during the observation period. By reducing bilateral percutaneous venous-layer penetrations from six to two while maintaining dose and treatment-territory equivalence, the single-point intramuscular technique may represent a structurally coherent refinement of established BoNT-A treatment concepts for lateral canthal rhytids. Because this was an uncontrolled preliminary clinical observation, the observed rate should be interpreted as descriptive only; any potential reduction in hematoma risk remains hypothetical.

Toxins doi: 10.3390/toxins18080327

Authors: Alessandro Picelli Rita Di Censo Ilaria Di Maria Antonella Vangelista Maria Vittoria Benetti Nicola Smania Valentina Varalta Mirko Filippetti

Cerebral palsy is a leading cause of childhood motor disability, and ambulatory children with spastic hemiplegia commonly present with dynamic equinus, calf muscle overactivity, and gait impairment. Evidence on focal vibration after botulinum toxin type A injection remains limited. This exploratory proof-of-concept pilot study assessed short-term changes following a four-week multimodal rehabilitation program incorporating focal vibration. In this uncontrolled single-group pre–post study, nine ambulatory children (mean age, 12.1 years), all classified as level II on the Gross Motor Function Classification System, received onabotulinumtoxinA injections into the affected gastrocnemius muscles, followed by twelve outpatient sessions combining focal vibration, conventional physiotherapy, and robotic gait training. Passive ankle dorsiflexion increased, calf muscle spasticity decreased, and selected gait parameters improved. Walking speed increased from 0.79 to 0.90 m per second, an absolute change of 0.11 m per second, comparable in magnitude to published estimates of clinically important change in ambulatory children with cerebral palsy. After Bonferroni correction, only walking speed and affected-limb stride length remained significant. No adverse events occurred. These findings describe outcomes associated with the combined program but do not establish the efficacy or independent contribution of focal vibration.

Toxins doi: 10.3390/toxins18080326

Authors: Juan Chen Bing Wang Yingying Xie Fei Xue Li Shi Yang Jiao Yongqiang Lin

Toad poison (ChanSu), a traditional animal-derived medicine, has long been used in East Asian medical systems for the treatment of inflammatory conditions and tumor-related disorders. While bufadienolides have been extensively investigated as its major bioactive constituents, the contribution of peptide components to its antitumor effects remains largely unexplored. Here, we identified protein-derived candidate antiproliferative peptides from toad poison using an integrated proteomics and bioinformatics-guided strategy. Proteomic analysis identified 135 proteins, from which 2117 peptide sequences were generated via in silico digestion with trypsin and pepsin. Subsequent multi-step screening using PeptideRanker, AntiCP, iACP, and ACPred yielded twelve candidate peptides with predicted anticancer activity. Network pharmacology analysis suggested their potential involvement in cancer-related targets and pathways. ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) evaluation was subsequently used as a complementary assessment of drug-like and safety-related properties, further prioritizing four peptides for experimental validation, while molecular docking supported their interactions with key lung cancer-associated targets. In vitro assays demonstrated that three of the four prioritized peptides (WEAWN, NSQWG, and ACGVIGICQ) exhibited initial antiproliferative activity against human lung cancer A549 cells, though these findings should be interpreted with caution given the absence of a positive control in the MTT assay. This study provides preliminary evidence suggesting that protein-derived peptide candidates from toad poison may possess antiproliferative potential, representing an early systematic exploration of its previously unexplored peptidome as a source of candidate antiproliferative peptides warranting further pharmacological investigation. The integrated strategy presented here offers an efficient approach for the discovery of bioactive peptides from animal-derived traditional medicines.

Toxins doi: 10.3390/toxins18080325

Authors: Georgios I. Papakonstantinou Christos Eliopoulos Dimitrios Arapoglou Nikolaos Tsekouras Katerina Manolakou Labrini V. Athanasiou Dimitrios Gougoulis Soultanidis Aris Vasileios G. Papatsiros

Mycotoxin contamination of cereal-based pig feeds is a persistent threat to swine health and global pork safety. Pigs are uniquely susceptible to mycotoxicosis, and the principal toxins—aflatoxins (AFs), deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FUMs), and ochratoxin A (OTA)—cause impaired growth, reproductive failure, hepato- and nephrotoxicity, immunosuppression, and carry-over residues in edible tissue. Anti-mycotoxin feed additives, including mineral adsorbents, biotransforming agents, and multicomponent mycotoxin-detoxifying agents (MMDAs) combining clays, phytogenic antioxidants (curcumin, silymarin), and postbiotics (yeast cell wall, hydrolyzed yeast), constitute the primary mitigation strategy. Beyond animal health, these agents play underappreciated roles in both the circular economy (CE)—enabling safe valorization of by-product feed ingredients and reducing feed waste—and in the life cycle assessment (LCA) sustainability profile of pork production, where feed conversion ratio (FCR) drives 22–95% of greenhouse gas emissions. Field-based in vivo studies in sows and weaned piglets, integrating plasma oxidative stress biomarkers (TBARS, protein carbonyls, total antioxidant capacity) with performance and reproductive endpoints, provide the most comprehensive real-world evidence for MMDA efficacy to date. This review synthesizes mycotoxin toxicology, anti-mycotoxin agent mechanisms, clinical evidence, and the CE/LCA sustainability framework, proposing integrated mycotoxin management as a One Health imperative in modern swine production.

Toxins doi: 10.3390/toxins18080324

Authors: Xingsai Liu Fengguang Zhao Kashif Iqbal Sahibzada Yaping Zhang Shan Wei Pingping Tian Yingying Wang Yuansen Hu Yangyong Lv

In this study, a previously uncharacterized zearalenone lactone hydrolase, ZHD30L, from Knufia peltigerae was identified and characterized, and semi-rational design was employed to enhance its thermostability. The results indicate that the optimal reaction conditions for this enzyme are 40 °C and pH 9.0, under which it catalyzes the ring-opening of the ZEN lactone to generate the non-toxic product HZEN. The mutant with five combined sites Mut5, (E11T-A127V-I180K-H235P-V156I), was obtained through semi-rational design combined with multiple rounds of iterative superposition and negative elimination strategies. Thermodynamic characterization revealed that, compared to the wild type, the t1/2 of Mut5 at 48 °C was significantly extended from 1.67 min to 796.8 min, and the Tm value increased from 39.56 °C to 50.04 °C. The markedly prolonged thermal half-life and increased apparent melting temperature may improve the operational robustness of Mut5 during moderate-temperature feed and food processing or post-processing applications, potentially reducing activity loss and the need for repeated enzyme additions. Concurrently, the kcat/KM improved from 19.08 μM−1·s−1 (95% Cl = 12.80 to 25.36 μM−1·s−1) to 33.29 μM−1·s−1 (95% Cl = 23.20 to 43.38 μM−1·s−1). Under optimal reaction conditions, WT and Mut5 degraded 76.51% ± 1.65% and 99.13% ± 0.71% of ZEN (10 μg/mL), respectively, within 3 min at equal enzyme concentrations. Molecular dynamics simulations indicate that the H235P and E11T mutations reduce local conformational entropy, the A127V and V156I mutations optimize internal hydrophobic stacking, and the I180K mutation reshapes the local charge microenvironment. Furthermore, Mut5 exhibited an overall RMSD at high temperatures that was 0.026 Å lower than that of the wild type, and a global free energy minimum that was 0.36 kcal/mol lower. These computational results offer possible structural interpretations, providing a theoretical reference for developing highly efficient enzyme preparations for the feed and food industries.

Toxins doi: 10.3390/toxins18080323

Authors: Waleska Stephanie da Cruz Nizer Giulliana Altaf dos Santos William Gustavo Lima Felipe Henrique de Souza Silva Wanderson Aparecido Brandão Candido Amanda Neves de Souza Giovanna Paula Araújo Rodrigo Moreira Verly Maria Elena de Lima

The emergence of multidrug-resistant (MDR) bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA), represents a major global health challenge by limiting the current therapeutic options. In this context, antimicrobial peptides (AMPs) have been widely studied for their potent antimicrobial properties. In this study, we evaluated the anti-MRSA effect of a novel AMP, LyeTx I mnΔKL, derived from a toxin of Lycosa erythrognatha. Its activity was evaluated in vitro by minimal inhibitory and bactericidal concentrations (MIC and MBC), antibiofilm effect, membrane interaction, cytotoxicity, synergistic interaction with vancomycin, and in vivo in an MRSA murine wound/abscess infection model. LyeTx I mnΔKL showed enhanced antimicrobial activity against clinical MRSA isolates compared to its prototype (LyeTx I mnΔK), with MIC50 and MBC50 of 2 and 8 µM and 16 and 32 µM, respectively. Furthermore, LyeTx I mnΔKL exhibited a rapid bactericidal effect and a pronounced ability to inhibit biofilm formation and disrupt mature biofilms. LyeTx I mnΔKL interacts with bacterial membranes, adopts an α-helical structure, and induces membrane disruption and leakage of intracellular material. In vivo, topical treatment with LyeTx I mnΔKL reduced MRSA burden compared with LyeTx I mnΔK and untreated controls (log10 CFU/g of wound of 2.4, 4.4, and 6.5 for LyeTx I mnΔKL, LyeTx I mnΔK, and the saline group, respectively). However, increased cytotoxicity remains a significant limitation. Overall, LyeTx I mnΔKL is a promising anti-MRSA candidate for topical use with potent antibiofilm and in vivo activity.

Toxins doi: 10.3390/toxins18080322

Authors: Upasana Dhakal Christopher Toomajian

Fusarium head blight (FHB) caused by Fusarium graminearum is a major disease of wheat and barley worldwide. Besides causing yield loss, F. graminearum also contaminates infected grains with trichothecene mycotoxins such as deoxynivalenol (DON) and its acetylated derivatives. Field isolates of F. graminearum vary in the amount of mycotoxins produced, both on infected wheat heads and in controlled laboratory experiments. Genes encoding the enzymes responsible for trichothecene mycotoxin biosynthesis are already characterized, but additional genes responsible for the variation in amounts of mycotoxins detected within and among populations remain to be identified. We measured levels of trichothecenes produced in vitro in a sample of 151 F. graminearum field isolates. Genome-wide association performed with these measurements identified 10 quantitative trait loci (QTL) associated with variation in DON and/or 15ADON levels. The candidate regions contain many functionally characterized genes, including the Swr1p helicase gene, an MFS transporter, and multiple other transmembrane transporters that may relate to the fungus’ ability to transport trichothecenes across membranes for sequestration or export. These results help to characterize the genetic factors that influence variability in trichothecene levels, which contribute to our understanding of trichothecene levels on infected grain and may lead to strategies to mitigate this toxin contamination.

Toxins doi: 10.3390/toxins18080321

Authors: Oscar Sánchez-Carranza Claudia Jatzke Andreas Gravius Jens Nagel

Schwann cells (SC) myelinate peripheral axons and orchestrate nerve regeneration after injury by switching between myelinating, proliferative and repair states. Evidence suggests that Botulinum Neurotoxin Type A (BoNT/A) influences SC biology, potentially supporting nerve repair and pain relief in peripheral neuropathic pain (PNP) models. However, BoNT/A receptor and target expression in human SC (hSC) remains poorly explored. Here, this translational gap was addressed by transcriptionally profiling genes encoding BoNT/A-relevant receptors and targets in primary hSC and testing whether paclitaxel evokes hSC phenotype plasticity in vitro based on changes in gene expression. Primary hSC were isolated, cultured, and treated with paclitaxel or vehicle, followed by RT-qPCR profiling of hSC markers and BoNT/A receptor/targets genes. Untreated hSC expressed moderate NGFR and S100β, with low MBP levels, suggesting a non-myelinating state profile. Transcripts encoding the BoNT/A receptor machinery (SV2A, SYT1) and the target SNAP25 were detectable at moderate levels. Paclitaxel induced changes in gene expression: SV2A and SYT1 decreased (up to two-fold), whereas SNAP25 and MBP increased, accompanied by reduced NGFR, indicating a shift toward a more differentiated transcriptional state. These data indicate hSC transcriptional plasticity in vitro and provide transcriptional evidence for the expression of BoNT/A-related molecular components in non-neuronal human cells.

Toxins doi: 10.3390/toxins18080320

Authors: Xia Cui Jing Zhang Zixiao Zhou Ziyi Wang Jing Hou Rong Zhao Sai Fan

Arachidonic acid (ARA) is an essential long-chain polyunsaturated fatty acid for infant growth and development, commonly added to infant formula and supplementary foods as ARA oil or ARA powder. Since December 2025, infant formula products have been recalled in several countries due to cereulide contamination in ARA ingredients. Cereulide, a heat-resistant emetic toxin produced by Bacillus cereus, presents a serious threat to infant health. However, specific analytical methods for cereulide detection in ARA raw materials remain scarce, hindering effective quality control and source prevention of food safety hazards. This study developed and validated a rapid ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) method for the quantitative determination of cereulide in ARA oil and powder. Sample pretreatment was optimized according to matrix characteristics: for microencapsulated ARA powder, water dissolution was performed to release cereulide before acetonitrile extraction, whereas direct extraction was applied to ARA oil. Subsequent HLB-P pass-through solid-phase extraction effectively eliminated matrix interference in both matrices. The method exhibited excellent linearity (R2 > 0.999) from 0.1 to 20 μg·L−1, with an LOD of 0.03 μg·kg−1 and an LOQ of 0.1 μg·kg−1, respectively. Spike recoveries reached 90.7–107.0% for ARA oil (RSDs ≤ 6.5%) and 88.5–107.9% for ARA powder (RSDs ≤ 7.0%). This rapid and reliable method provides a critical analytical tool to monitor cereulide in ARA raw materials and mitigate infant safety hazards at the source.

Toxins doi: 10.3390/toxins18080319

Authors: Diego Bagnasco Benedetta Bondi Greta Losacco Carola Montagnino Francesca Froio Elena Tedesco Gloria D’Alessandro Ilaria Baglivo Laura Bruno Sara Chiappori Maria José Murillo Jaramillo Marcello Mincarini Fulvio Braido Cristiano Caruso

Staphylococcus aureus frequently colonizes the skin and upper airways and produces a broad repertoire of immunomodulatory molecules. In asthma, the most consistent evidence concerns staphylococcal enterotoxins (SEs), which can act both as superantigens and as allergens, and IgE sensitization to SEs (SE-sIgE). SE-sIgE is associated with severe asthma, type 2 inflammation, chronic rhinosinusitis with nasal polyps (CRSwNP), exacerbations, and, in some longitudinal studies, persistent airflow obstruction. However, this relationship is not necessarily causal: SE-sIgE may reflect exposure, an immune response, or a biologically active endotype, whereas colonization, local toxin production, and systemic sensitization are not equivalent. SEs simultaneously bind class II MHC molecules and Vbeta regions of the T-cell receptor, activating large fractions of T lymphocytes; they also promote IL-4, IL-5, and IL-13 production, polyclonal B-cell activation, local IgE synthesis, mast-cell degranulation, eosinophilia, and IL-8/neutrophil circuits. Alpha-toxin (Hla) and SEB can damage the epithelial barrier, facilitating allergen penetration and alarmin signalling. These observations support an interaction model in which dysbiosis, barrier dysfunction, and type 2 immunity mutually reinforce one another along the nasobronchial axis. Corticosteroids and antibiotics may modify selected nodes in this circuit, but current evidence is insufficient to recommend decolonization or antitoxin therapy in stable asthma. Biologics interrupt downstream pathways potentially fuelled by toxins: omalizumab neutralizes free IgE; mepolizumab and benralizumab reduce the eosinophilic axis; dupilumab blocks IL-4/IL-13 signalling; and tezepelumab acts upstream on TSLP. Nevertheless, randomized trials stratified by SE-sIgE are lacking, and no evidence demonstrates that these treatments eliminate colonization or toxin production. SE-sIgE therefore appears to be a promising biomarker, particularly in severe asthma with CRSwNP, but it is not yet an autonomous criterion for biologic selection. A broader barrier-organ analysis also identifies nasal, cutaneous, and intestinal colonization as distinct ecological states; atopic dermatitis as a complementary model of toxin-amplified type 2 inflammation; and biofilms and extracellular vesicles as candidate mechanisms of persistent toxin delivery. These data increase biological plausibility but remain indirect for asthma.

Toxins doi: 10.3390/toxins18070318

Authors: Xiaohong Ge Zhikun Chen Haoyuan Guo Junjian Ran

Fusarium head blight (FHB) caused by Fusarium graminearum leads to huge yield losses and mycotoxin contamination in wheat globally. Paenibacillus polymyxa with strong antagonistic activity was preliminarily identified. To clarify the key antifungal component, an extracellular protein was purified via ammonium sulfate precipitation, DEAE-52 anion-exchange and Sephadex G-75 gel filtration chromatography. SDS-PAGE showed a single band at 76 kDa. liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis confirmed this protein belongs to glycosyl hydrolase family with 86% sequence coverage. Biochemical characterization showed that the crude protein was stable at 40–90 °C and pH 3.0–9.0, sensitive to proteinase K, trypsin and neutral protease. The purified 76 kDa protein exhibited antifungal activity against F. graminearum. The gene encoding this protein was cloned and expressed in Escherichia coli. The renatured recombinant protein p76kd showed comparable antifungal activity to the native protein. This study purified and characterized a 76 kDa protein annotated as a glycosyl hydrolase via LC-MS/MS peptide matching; its antifungal function is presumed to originate from the conserved glycosyl hydrolase domain according to existing homologous research, which is distinct from previously reported lipopeptides or uncharacterized complexes. This protein provides a promising candidate for the biocontrol of FHB and related fungal diseases in cereal crops.

Toxins doi: 10.3390/toxins18070317

Authors: Till-Alexander Plinkert Johanna Doll-Lee Edoardo Passarotto Christos I. Ioannou Eckart Altenmüller André Lee

Musician’s Dystonia (MD) is a task-specific movement disorder that occurs while playing an instrument and severely limits a musician’s ability to perform. While currently no causal therapies are available, symptomatic therapies, particularly the injection of botulinum toxin (BoNT), offer a therapeutic option. The present study presents a 30-year longitudinal evaluation of the clinical course of 156 patients with MD treated with BoNT at a specialist outpatient facility. The evaluation took into account known risk factors, such as family history, time to treatment, initial severity of symptoms, gender, and age at the onset of playing an instrument and the onset of symptoms. The study showed that more severe initial symptoms, which entailed a larger number of muscles requiring treatment, predicted a poorer long-term outcome. Furthermore, a longer interval between symptom onset and the commencement of BoNT treatment was also associated with a poorer outcome. These results highlight the importance of rapid diagnosis and the early initiation of BoNT therapy in MD patients.

Toxins doi: 10.3390/toxins18070316

Authors: Edward Agyarko Omeralfaroug Ali Lucy Ikanya Attila Zsarnovszky Melinda Kovács András Szabó

Fumonisins, a class of mycotoxins produced primarily by Fusarium fungi, pose significant health risks to humans and animals through contamination of the food and feed chains. They rank among the most prevalent mycotoxins contaminating maize and maize-derived feeds worldwide, resulting in chronic dietary exposure of both humans and livestock populations across many regions. Their core mechanism of action is the inhibition of ceramide synthases (CerS), which disrupts the essential balance of sphingolipid metabolism by causing an accumulation of free sphingoid bases and a depletion of complex sphingolipids. Both sphingolipids and steroidogenesis are metabolically linked to mitochondrial, membrane and kinase-cascade mechanisms; hence this metabolic disruption may consequently affect steroid hormone biosynthesis, triggering toxicity phenotypes marked by impaired gametogenesis hormonal imbalances, and compromised pregnancy outcomes across mammalian species. Despite the established link between fumonisins and sphingolipid disruption, there is a gap in the literature, as no study to date has integrated sphingolipid disruptions with steroid hormone levels in a dose-dependent manner within reproductive tissues in vivo. This review synthesizes current scientific knowledge across mammalian species to highlight the risks fumonisins pose to reproductive physiology and to identify directions for future research.

Toxins doi: 10.3390/toxins18070315

Authors: Yingyi Wu Shuying Lin Chuhan Shao Cheng Zhang Kaiyin Xie Kaizhao Zhang Xiaohong Huang Hongjie Cui

Ochratoxin A (OTA) is a widespread nephrotoxic mycotoxin that contaminates cereal grains and feed, posing serious threats to animal health and food safety. Although curcumin has been reported to exert protective biological activities, it remains unclear whether its protection against OTA-induced renal epithelial injury is mediated by broad transcriptomic reversal or by selective regulation of key survival pathways. This study investigated the protective effect of curcumin against OTA-induced apoptosis in porcine renal epithelial (PK-15) cells and explored the underlying molecular mechanism using transcriptome sequencing combined with molecular validation. PK-15 cells were treated with 8 μg/mL OTA (approximately 19.8 μmol/L), 10 μmol/L curcumin, or their combination. Cell viability, LDH release, apoptotic morphology, mitochondrial membrane potential (ΔΨm), and apoptosis rate were assessed by CCK-8, LDH assay, Hoechst 33342 staining, JC-1 staining, and flow cytometry. Transcriptome sequencing was performed to identify global gene expression changes and key signaling pathways, followed by qRT-PCR and Western blot validation of apoptosis-related factors and the PI3K/AKT/mTOR pathway. OTA markedly reduced cell viability, increased LDH release, induced nuclear condensation and apoptotic body formation, and decreased ΔΨm. Transcriptome analysis revealed that OTA caused extensive transcriptional dysregulation (11,707 differentially expressed genes), whereas curcumin selectively modulated 498 genes, of which 380 overlapped with OTA-responsive genes. KEGG enrichment identified the PI3K-Akt signaling pathway as a key regulatory target. At the mRNA level, OTA upregulated Bax and Caspase-3 and downregulated Bcl-2; corresponding changes were observed at the protein level, and curcumin reversed these effects. Furthermore, OTA reduced the mRNA levels of PI3K, AKT, and mTOR and the protein abundance of PI3K, p-AKT, and mTOR, whereas curcumin partially restored these changes. In conclusion, curcumin alleviates OTA-induced mitochondrial apoptosis in PK-15 cells mainly by restoring PI3K/AKT/mTOR-associated survival signaling and Bcl-2/Bax/Caspase-3 balance rather than by broadly reversing the entire transcriptomic disturbance. These findings provide mechanistic insight into curcumin-mediated protection against OTA-induced renal epithelial toxicity.

Toxins doi: 10.3390/toxins18070314

Authors: Artur Drużdż Małgorzata Dudzic Joanna Poszwa Igor Bednarski Katarzyna Hojan Jolanta Dorszewska

Cervical dystonia (CD) is a focal dystonia typically treated with botulinum toxin type A (BoNT-A), but objective biomarkers of disease state and treatment response are limited. In a prospective controlled observational study with within-subject phase comparison, we evaluated clinical, neurophysiological, and biochemical markers at two time points (waning and peak response phases) within the BoNT-A injection interval in 30 CD individuals (with caput, collis and mixed patterns) and 25 healthy controls. We measured clinical status (TWSTRS and VAS pain), neurophysiology (F-wave minimal latency (F-min) and cutaneous silent period (CSP) indices) and plasma biochemical markers (neurofilament light chain (NfL) and zonulin). Compared with controls, CD patients showed lower NfL levels, higher zonulin, shorter CSP end duration and shorter CSP-derived central conduction time (CSP-CCT); nerve conduction velocity and CSP onset were similar. From waning to peak, TWSTRS (all subscales) and VAS improved, zonulin decreased, NfL increased, CSP end duration and CSP-CCT increased, and Fmin slightly prolonged. Changes in CSP measures correlated with pain (VAS and TWSTRS pain), while zonulin changes showed no significant correlations and NfL changes correlated weakly/negatively with CSP end time. Phase-dependent changes in biochemical and neurophysiological markers were observed alongside clinical measures across the BoNT-A treatment cycle in CD, suggesting potential associations between peripheral biochemical markers, central inhibitory measures, and treatment phase. These findings may warrant further investigation of multimodal biomarker approaches in CD.

Toxins doi: 10.3390/toxins18070313

Authors: Sarra Rafai Lara Manyes Ana Moreno Alessandra Cimbalo Giuseppe Meca Victor Dopazo

Contamination of food and feed by aflatoxin B1 (AFB1) remains a major global concern due to its toxicity, carcinogenicity, and persistence in the food chain. Environmental and climatic pressures continue to favor aflatoxigenic fungal contamination, highlighting the need for effective and sustainable detoxification strategies. Biological detoxification has emerged as a promising alternative to conventional physical and chemical treatments. This systematic review, conducted following PRISMA guidelines, summarizes microbial and enzymatic approaches for AFB1 detoxification, focusing on bacteria, yeasts, and microbial enzymes. The literature shows a predominance of bacterial systems, especially lactic acid bacteria and Bacillus species, mainly acting through adsorption, fungal growth inhibition, and suppression of aflatoxin biosynthesis. Yeasts, although less represented, also showed promising detoxification capacities through adsorption and biodegradation-related mechanisms. Enzymatic systems achieved the highest efficiencies, particularly oxidative enzymes such as laccases and dye-decolorizing peroxidases, often exceeding 90% detoxification under optimized conditions. However, industrial application remains limited by laboratory-scale validation, variability among protocols, incomplete toxicological assessment of degradation products, and limited evidence in complex food and feed matrices.

Toxins doi: 10.3390/toxins18070312

Authors: Stefano Stuard Charles Hugh-Jones Dinesh Chatoth Michael Anger Alfred Gagel Bernard Canaud

Uremic syndrome results from the accumulation of biologically active solutes that contribute to inflammation, oxidative stress, endothelial dysfunction, cardiovascular disease, anemia, CKD–mineral and bone disorder, and protein-energy wasting in patients with end-stage kidney disease. While conventional hemodialysis efficiently removes small water-soluble compounds, clearance of middle molecules and protein-bound uremic toxins remains limited. Post-dilution hemodiafiltration (HDF) combines diffusion and convection to enhance removal across a broader molecular-weight spectrum. This review summarizes the mechanisms of uremic toxin accumulation, the biologic and clinical relevance of major retained solutes, and current evidence supporting improved toxin removal with HDF. Randomized trials, meta-analyses, and observational studies have associated HDF with reduced inflammatory burden, improved hemodynamic tolerance, and favorable clinical outcomes, supporting its role as an advanced strategy for targeting the broader spectrum of uremic toxicity.

Toxins doi: 10.3390/toxins18070311

Authors: Regiane Stafim da Cunha Paulo Cézar Gregório Amanda Carina Coelho Morais Fellype Carvalho Barreto Andréa Emilia Marques Stinghen

Chronic kidney disease (CKD) represents a significant public health problem. Patients with CKD have high morbidity and mortality, mainly due to cardiovascular diseases. As CKD progresses, impaired renal clearance leads to the progressive accumulation of uremic toxins, culminating in deleterious effects on multiple organ systems. Uremic toxins contribute to cardiovascular disease by inducing inflammation, oxidative stress, and endothelial barrier dysfunction, thereby promoting atherosclerotic processes. They are also potentially implicated in neurological impairment, bone disorders, and sarcopenia. Some uremic toxins originate from dietary components and gut microbiota metabolism, which are key mechanisms and pathways in uremic toxicity. Current dialysis therapies only partially mitigate uremic toxicity, highlighting the need for strategies that reduce uremic toxin generation as well as more efficient dialysis modalities. Given the importance of uremic toxins in CKD progression, this review provides a comprehensive overview of uremic toxicity, focusing on its pathophysiological mechanisms, its relationship with the gut microbiota, and emerging interventions to mitigate its deleterious effects.

Toxins doi: 10.3390/toxins18070310

Authors: Jacopo Gardellin Marta D’Angelo Lorenzo Spadotto Giacomo Checchin Giovanni Gaetti Luca Gambolò Giuseppe Stirparo Nicola Merli

Background: Increasing evidence supports the existence of a somatosensory subtype of tinnitus, in which craniocervical–craniomandibular structures can modulate tinnitus perception also through central auditory pathways. Despite growing mechanistic evidence, no targeted pharmacological intervention has been validated for this subtype. Objectives: This study aimed to synthesize the anatomical and neurophysiological rationale linking masseter hyperactivity to somatosensory tinnitus and to discuss BoNT/A as a hypothesis-generating therapeutic option requiring clinical validation. Methods: A PRISMA-ScR-guided scoping review was conducted in MEDLINE and EMBASE, using three complementary search lines (last search: 31 May 2026). Results: Of 284 records screened, four studies met eligibility criteria: three from the search strategy on BoNT/A and tinnitus and one from the search strategy on tinnitus and masticatory dysfunction. One additional record (Ranoux and Levine, 2024) was identified through manual search, describing tinnitus improvement following periauricular and splenius capitis BoNT/A injections, with masseter/temporalis injection proposed as an alternative site in the same protocol. Conclusions: Current anatomical and neurophysiological evidence supports a plausible link between masseter hyperactivity and somatosensory tinnitus in selected patients. BoNT/A is established for reducing masseter hyperactivity and may theoretically reduce abnormal somatosensory input; however, direct clinical evidence for tinnitus improvement remains insufficient. Its use should be considered experimental or hypothesis-based, within a multidisciplinary framework and with standardized outcome assessment. Prospective controlled trials are needed.

Toxins doi: 10.3390/toxins18070309

Authors: James S. Metcalf Manuel Aparicio Sandra A. Banack Jason Pim John R. Cassani Paul A. Cox

Cyanobacterial and harmful algal blooms are common components of the waters in and around Southwest Florida and are sufficiently frequent to be of concern with respect to adverse effects on short- and long-term human and animal health. Currently, the contribution of airborne exposure to cyanobacterial toxins is not as advanced as for the other known human exposure routes such as consumption of contaminated water or fish. An airborne monitoring device named Airborne Detection for Algae Monitoring (ADAM) was developed to collect air samples in the proximity of algal and/or cyanobacterial blooms to examine the relationship between naturally occurring toxins in the air and those in proximal Floridian waters. Twenty-one air and water samplings were performed between July and December 2021 using a filter and impinger system for airborne components, along with a water sample from the same location. Samples were assessed for microcystins, anatoxin-a, cylindrospermopsin, saxitoxin and BMAA and isomers. Furthermore, as Karenia brevis is common in this area, brevetoxins were also assessed in air samples. Although very few large-scale cyanobacterial bloom events were observed at sampling locations, cyanobacterial and algal toxins were present in the majority of samples at low concentration. Data obtained from ADAM indicate that people may be chronically exposed to low concentrations of cyanobacterial toxins and that further assessment is required to help protect human health.

Toxins doi: 10.3390/toxins18070308

Authors: Yongping Jiang Bowen Tai Yufan Yang Xinyue Zhang Jing Jin Fuguo Xing

Aflatoxin B1 (AFB1) contamination in maize poses serious risks to food and feed safety; however, conventional laboratory-based assays are often constrained by time and throughput for large-scale screening. This study proposes a hyperspectral imaging (HSI) workflow for rapid, non-destructive prediction of AFB1. A total of 236 hyperspectral images were acquired in the 400–1000 nm range (256 bands) from maize samples covering a broad gradient of AFB1 contamination, and spectral features were extracted from regions of interest (ROI) for model development. The results demonstrate that appropriate spectral preprocessing and wavelength selection play a critical role in improving model robustness, with the SNV–CARS–KNN model achieving the best prediction performance (test R2 = 0.9341, RMSE = 4.8938). Based on the predicted AFB1 values, contamination grading was further explored to enable rapid screening and risk management in agricultural applications. Aflatoxin B1 (AFB1) contamination in maize poses substantial risks to food and feed safety, creating a need for rapid screening tools that can support high-throughput prescreening. In this study, hyperspectral imaging (HSI) was explored as a non-destructive approach for the preliminary assessment of AFB1 contamination in maize kernels. A total of 236 hyperspectral images were collected in the 400–1000 nm range, and region-of-interest spectra were extracted for model development. Spectral preprocessing and wavelength selection strategies were compared in combination with several conventional regression models to examine their influence on predictive performance using the current dataset. Among the tested combinations, the SNV–CARS–KNN model showed the most favorable performance on the held-out test set (R2 = 0.9341; RMSE = 4.8938). In addition, a grade-based classification derived from predicted values was explored as an application-oriented extension for rapid risk sorting. However, the study was conducted on artificially contaminated samples and relied on rapid-test-derived reference values, so the findings should be interpreted as a proof of concept rather than a fully validated quantitative method. Overall, the results support the potential of HSI for rapid prescreening of AFB1 in maize and provide a basis for further validation using more rigorous reference analysis and independent sample sets.

Toxins doi: 10.3390/toxins18070307

Authors: Kohei Kazuma Naoki Tani Katsuhiro Konno Migaku Kawaguchi Hidetoshi Inagaki

Digger wasps are solitary apoid wasps that excavate ground nests and use venom to paralyze insect prey. Recent phylogenetic analyses suggest that digger wasps belong to an ancient lineage of aculeate Hymenoptera and share an evolutionary ancestry with other venomous hymenopterans, including bees and ants. Although the venoms of ants, bees, and social wasps have been extensively studied, those of digger wasps remain poorly characterized in terms of their composition, molecular diversity, and biological activity. In this study, we investigated the venom of the digger wasp Cerceris japonica using integrated transcriptomic and proteomic analyses. We identified 19 toxin-like proteins and peptides, 14 non-toxin-associated components, and 11 novel peptides and proteins with no detectable similarity to known peptides and proteins. Among these, peptide Cj 2 exhibited insecticidal activity but showed no antimicrobial, hemolytic, or nicotinic acetylcholine receptor-modulating activities. These findings provide new insights into the molecular diversity and biological activities of digger wasp venom, expand our understanding of venom evolution in venomous hymenopterans, and serve as a framework for elucidating the conserved and lineage-specific features of hymenopteran venoms.

Toxins doi: 10.3390/toxins18070306

Authors: Matteo Val Ludovica Zucchini Matteo Pollis Luca Guarda-Nardini Luca Lombardo Daniele Manfredini Anna Colonna

Myofascial pain of the masticatory muscles, frequently associated with bruxism, is a prevalent condition with substantial impact on quality of life. Botulinum toxin type A (BoNT-A) is used as an adjunctive treatment for refractory cases, but the temporal pattern of masseter morphometric change after injection remained incompletely characterised. This prospective observational pilot study was designed and reported strictly as a 3D imaging and morphometric methodology study; clinical pain and patient-reported outcomes were outside its scope and were not assessed. Eleven adults with myofascial pain and bruxism received BoNT-A (100 U total: 30 U per masseter side, 20 U per anterior temporalis side). Structured-light 3D facial scans (EinScan H, Shining 3D) were acquired at baseline (T0), one month (T1), and three months (T2). A standardised masseter region of interest (“TGL area”) was defined using cutaneous landmarks (tragus, gonion, labial commissure). Measurement precision was confirmed by intra-session repeatability (0.083 ± 0.020 mm), registration accuracy (RMS error 0.189–0.200 mm), and excellent landmark reliability (mean ICC [3, 1] = 0.955, 95% CI: 0.843–0.991). Repeated-measures ANOVA showed a significant effect of time on bilateral surface displacement (F(1.85,18.52) = 14.39, p < 0.001, η2G = 0.44), with significant reductions at T1 (–0.716 ± 0.480 mm; p = 0.002, Hedges’ g = 2.03) and T2 (–0.847 ± 0.588 mm; p = 0.002, g = 1.96) relative to baseline, and no significant change between T1 and T2 (p = 1.000). Changes were bilaterally symmetric (symmetry index at T1: 0.231 ± 0.154 mm). These findings indicate that 3D structured-light scanning reliably detects subtle facial contour changes after BoNT-A injection and may complement clinical and patient-reported assessments; the pilot design and small sample preclude definitive conclusions.

Toxins doi: 10.3390/toxins18070305

Authors: Vinícius Moura Mário Lopes Lucas Butturi Barbara Doupovec Giovana Longhini Marcelo Viviani Fabricia Roque Vitor Pais Lúcio Araújo Cristiane Araujo

Mycotoxins are a worldwide hazard to animal feed and capable of deleterious effects on poultry. The present study aimed to evaluate the effects of an anti-mycotoxin feed additive on productive and reproductive parameters of breeder hens and roosters fed diets experimentally contaminated with zearalenone (ZEN). A total of 288 hens and 32 roosters (Cobb 500) were randomly distributed in one of four dietary treatments, with nine hens and a rooster per pen and eight pens per treatment: control (C); C + anti-mycotoxin (AM) additive; C + ZEN; and C + AM + ZEN. The trial lasted from 28 to 65 weeks of age. Egg production, egg quality, fertility, hatchability, and hatch rates of fertile eggs were evaluated, as were sperm morphology and abnormalities, and chick quality. ZEN inclusion did not affect egg production but had harmful effects on albumen, egg quality and weight, eggshell thickness, egg fertility and hatchability, and chick length and quality, compared to C, with this negative effect partially alleviated by AM inclusion. Both AM and ZEN, however, decreased spermatic concentration, without affecting sperm morphology or abnormality. The anti-mycotoxin additive effectively mitigated the deleterious effects of ZEN on reproductive parameters and egg quality indexes when fed to breeder hens.

Toxins doi: 10.3390/toxins18070304

Authors: Kazuya Yoshida

Botulinum neurotoxin (BoNT) injection into the masseter muscle is used in selected conditions associated with excessive or painful masseter activity, including oromandibular dystonia, masseter-related myalgia, and bruxism-associated masticatory muscle pain. However, clinical outcomes and adverse-event profiles vary widely, partly because injection technique, dose, dilution, injection depth, anatomical targeting, patient selection, and injector experience have not been consistently standardized. This review examines the anatomical and pharmacological rationale for motor endplate zone (MEZ)-targeted BoNT injection into the masseter muscle. MEZ-targeted injection is not presented as an established clinical guideline or as a proven superior technique but as a hypothesis-generating framework for future controlled studies. Anatomical and electrophysiological studies indicate that neuromuscular junctions in the masseter muscle are concentrated within relatively restricted MEZs. From a pharmacological perspective, delivery of BoNT close to these zones may allow more efficient neuromuscular blockade with lower doses, although direct clinical evidence demonstrating superiority over conventional techniques remains insufficient. Reported adverse events may be influenced by excessive dosing, inaccurate injection depth, diffusion into adjacent structures, or repeated high-dose injections. Future studies should evaluate anatomically informed, dose-minimized injection strategies using clearly defined clinical populations, standardized diagnostic criteria, detailed reporting of injection technique, objective functional outcomes, and systematic adverse-event assessment.

Toxins doi: 10.3390/toxins18070303

Authors: Slobodan Dolašević Nikola Rokvić Marko Jauković Maja Petričević Tanja Keškić Marija Gogić Aleksandar Ž. Kostić

Bee-collected pollen (BCP), due to its valuable nutrient content, is considered a natural food suitable for human consumption. On the other hand, it should be noted that the rich nutritional value of BCP is also relevant in the context of potential contamination with fungal secondary metabolites, such as mycotoxins. In this study, a total of 26 pollen samples were collected from different locations in order to investigate the potential presence of mycotoxins. All BCP samples were obtained from beekeepers who produce pollen for further commercialization. The analyzed mycotoxins included ochratoxin A (OTA), aflatoxin B1 (AFB1), zearalenone (ZEN), T-2 toxin, HT-2 toxin, deoxynivalenol (DON), and fumonisins (FUMs), which were measured by direct competitive enzyme-linked immunosorbent assays. The obtained results indicated the presence of at least three toxins in all samples. AFB1, OTA, and ZEN were detected above the limit of quantification (LOQ) in all samples, whereas T-2 and HT-2 were below the LOQ in five samples and DON in four samples. FUMs were below LOQ in all samples. Due to the significant content of AFB1 determined in all samples, there is a potential risk for future consumers, demanding constant monitoring as well as improvement of BCP production.

Toxins doi: 10.3390/toxins18070302

Authors: Hyun Jin Park You-Jin Choi

The posterior belly of the digastric muscle (PBDM) has recently been suggested as a target for botulinum toxin (BoNT) treatment in facial synkinesis. However, anatomically based injection sites for the PBDM have not been established. This study aimed to identify an ideal BoNT injection site by analyzing the intramuscular nerve distribution of the PBDM and its anatomical relationships with surrounding structures. Twelve PBDM specimens were obtained bilaterally from six embalmed cadavers. Modified Sihler’s staining was performed to visualize intramuscular nerve distribution. The PBDM was divided into anterior, middle, and posterior thirds, and the course and terminal branches of the facial nerve were analyzed. In all specimens, the PBDM was innervated by a single branch of the facial nerve. Nerve entry points were located in the middle third in 75% of cases and in the posterior third in 25%. Intramuscular nerve endings were most densely distributed in both the anterior and middle thirds in all specimens, whereas the posterior third showed a lower distribution (25%). The anterior and middle thirds of the PBDM appear to be the most appropriate targets for BoNT injection. The area 1 cm posterior to the angle of the mandible may serve as a practical landmark-guided injection site in clinical practice.

Toxins doi: 10.3390/toxins18070301

Authors: John McGuire Karin Goodfriend Sarah Golus Whitney Morelli Mary Elizabeth S. Nelson-Biersach Nicholas Ketchum

Objective: Residency programs in the United States lack the necessary training for physicians to confidently and competently inject botulinum neurotoxin (BoNT) and manage complex spasticity patients. This creates an unmet need for spasticity patients who could benefit from BoNT injections. This study measured the impact of additional BoNT injection and spasticity management training on post-residency medical practice. Design: Graduates from our Physical Medicine and Rehabilitation residency from 2008 to 2024 were surveyed. Responses were collected from physicians who had residency BoNT injection and spasticity management training (RT) and compared to fellowship BoNT injection and comprehensive spasticity management training (FT). Results: 15 RT and 14 FT physicians completed the survey and were evaluated using descriptive statistics. Spasticity and dystonia treatments comprised an average of 10% of the RT and 62% of the FT clinical practice. BoNT injections involved 5% of the RT practice and 52% of the FT practice. Conclusion: Disabling spasticity is undertreated in this country, partly due to a shortage of training programs for physicians to become skilled and confident at injecting BoNT and comprehensive spasticity management. FT programs can help meet this unmet need by increasing the number of physicians who can provide specialized BoNT injections and spasticity management.

Toxins doi: 10.3390/toxins18070299

Authors: Hanke Zhao Changfeng Xiao Hui Jiang Lihui Zhu Junhua Yang Fang Gan

Mycotoxin contamination in feed poses a major threat to animal health and productivity, particularly in pigeons. This study evaluated the protective effects of three mycotoxin biodegradation agents (MBAs) in pigeons subchronically co-exposed to a mycotoxin-contaminated (MYC) diet for 60 days. The MYC diet was prepared by mixing 3% naturally mold-contaminated corn with 97% basal diet, yielding final dietary concentrations of fumonisin B1 (7829.41 µg/kg), aflatoxin B1 (201.16 µg/kg), zearalenone (1675.46 µg/kg), T-2 toxin (7721.41 µg/kg), and deoxynivalenol (1211.77 µg/kg). Birds were allocated to five groups (n = 18/group): CON (basal diet); MYC (MYC diet alone); and TRT1–TRT3 fed the MYC diet supplemented with 1.5 g/kg of MBA I (Bacillus subtilis + Bacillus licheniformis + Devosia sp.), MBA II (Bacillus licheniformis + Saccharomyces cerevisiae), or MBA III (Bacillus subtilis + glucose oxidase), respectively. Subchronic co-exposure to the MYC diet significantly increased serum pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), whereas all three MBAs effectively attenuated these elevations. Histopathological examination revealed inflammatory infiltration and tissue damage in the jejunum and liver of MYC birds, which were markedly alleviated by MBA supplementation. Gut microbiota analysis showed that mycotoxins decreased Firmicutes and Lactobacillus and increased Proteobacteria and Streptococcus, whereas MBAs partially restored microbial balance. Among the three MBAs, MBA II exhibited the strongest protective effect, as reflected by the greatest improvements in serum biochemistry, intestinal morphology, and microbial composition. Liver transcriptome analysis showed that mycotoxin mainly affected pathways related to cell cycle and DNA replication, whereas MBA II enhanced pathways associated with intestinal immune function and antigen processing and presentation. Collectively, these findings suggest that MBA II offers effective protection against combined mycotoxin-induced toxicity in pigeons and may serve as a promising feed additive for mitigating mycotoxin contamination in poultry production.

Toxins doi: 10.3390/toxins18070300

Authors: Melissa Regina Fessel Ana Marisa Chudzinski-Tavassi Fernanda Faria

Cystatins from hematophagous organisms modulate host proteases and are promising biotechnological tools. CysHv, a secreted cystatin from the Brazilian leech Haementeria vizottoi, was previously characterized as a potent inhibitor of papain and cathepsin L. In this work, it is shown that its inhibitory activity is tightly linked to conformational state. CysHv is purified as an active monomer but forms inactive dimers in a time-, temperature-, and concentration-dependent manner. A domain-swapping-like mechanism was hypothesized, and a disulfide-engineered mutant restricting local flexibility remained predominantly monomeric, supporting the role of conformational plasticity. Dimer formation correlates with loss of inhibitory activity and is accelerated at acidic pH, suggesting that pH modulates the kinetics rather than triggering dimerization. Comparative structural analysis of cystatins from hematophagous organisms revealed recurrent pairs of oppositely charged residues in β-strands flanking the inhibitory loop in secreted cystatins, but these pairs are generally absent in intracellular homologs. Although causality cannot be established and additional determinants likely contribute to dimerization, this pattern is consistent with pH-sensitive modulation of conformational behavior. Together, these findings identify CysHv as a dimer-prone cystatin for which oligomerization, potentially via a domain-swapping-like mechanism, could act as an environment-dependent regulator of inhibitory activity.

Toxins doi: 10.3390/toxins18070298

Authors: Flora Lefevre Rania Chermiti Julien Cebile Nathalie McKay Stanislas Bataille Stéphane Burtey Laetitia Dou

Patients with chronic kidney disease (CKD) are exposed to high levels of uremic toxins and have an increased risk of cardiovascular disease. Among these toxins, indolic compounds such as indoxyl sulfate (IS) are predictors of cardiovascular events and mortality in CKD patients and induce a procoagulant and proinflammatory vascular phenotype through activation of the aryl hydrocarbon receptor (AhR). Targeting AhR activation by indolic toxins may therefore help prevent cardiovascular complications in CKD. To this end, we investigated in vitro whether natural dietary AhR ligands (galangin, quercetin, curcumin and indole-3-carbinol) could antagonize IS-induced AhR activation and the associated inflammatory response in endothelial cells. The activation of the AhR genomic pathway was assessed by measuring the expression of AhR target genes (CYP1A1, CYP1B1, and AHRR) in endothelial cells and by evaluating AhR-dependent transcriptional activity using a CALUX-AHRE luciferase reporter assay in HG40/6 cells. In parallel, endothelial inflammation was evaluated by analyzing the expression of AhR-related inflammatory genes: F3/tissue factor, PTGS2/COX-2, CCL2/MCP-1, and CXCL8/IL-8. Quercetin was the only ligand capable of antagonizing IS-induced AhR transcriptional activity, as well as the upregulation of the endothelial AhR target genes CYP1A1 and CYP1B1. In contrast, galangin, curcumin, and I3C exhibited no inhibitory effects. Moreover, none of the tested dietary AhR ligands suppressed the IS-induced upregulation of endothelial inflammatory genes; instead, they tended to potentiate IS-induced inflammatory responses at high concentrations. In conclusion, among the AhR ligands tested, quercetin was the only one that attenuated IS-induced activation of the AhR genomic pathway in endothelial cells. However, it may also enhance IS-mediated endothelial inflammation, an effect also observed at specific concentrations of galangin, curcumin, and I3C. These findings suggest that the potential beneficial effects of natural dietary AhR ligands should be carefully considered in the context of CKD patients exhibiting high levels of indolic uremic toxins.

Toxins doi: 10.3390/toxins18070297

Authors: María Agustina Pavicich Andrea Patriarca

The landscape of mycotoxin research is evolving rapidly [...]

Toxins doi: 10.3390/toxins18070296

Authors: Carla de Sanctis Pecora Birgit Blessmann-Gurk Bianca Viscomi

The eyebrows are a central element of facial harmony, influenced by individual and external factors such as gender, ethnicity, facial shape, age, culture, and fashion trends. Traditional methods for esthetically enhancing eyebrows include plucking, highlighting, and surgical interventions. Another minimally invasive option for altering, thus enhancing eyebrow positioning, is botulinum toxin type A (BoNT-A) injection. The objective was to demonstrate a systematic, customizable approach for BoNT-A injections to reshape and reposition eyebrows according to individual facial shape enhancing balance. To determine optimal BoNT-A dose and injection points, an individualized assessment of facial and eyebrow shape and position was performed, leading to a customized protocol of neurotoxin injections according to each patient’s needs. The assessment method aims for the best match between the individual facial shape and eyebrow form and position. This technique enables individualized modulation of eyebrow position and shape, improving facial balance while addressing dynamic upper facial rhytides, thereby promoting harmonious outcomes that preserve individual facial characteristics and providing an additional approach to conventional eyebrow enhancement techniques.

Toxins doi: 10.3390/toxins18070295

Authors: Margaret-Jane Gordon Noshin Daud Louise Cantlay Silvia W. Gratz

The presence of Fusarium mycotoxins is an intractable problem in cereal production, with T-2 and HT-2 posing a particular issue in oats. This study assesses the prevalence of Fusarium mycotoxins in unprocessed and de-hulled oats and oat food products and quantifies the bioaccessibility of T-2, HT-2 and HT-2-glucoside from oat porridge in vitro. Twenty unprocessed food oat samples were de-hulled and mycotoxins quantified in unprocessed grains, hulls and groats using LC-MS/MS. Seventy-seven oat food products (40 porridge samples, 10 granola and muesli samples, 11 oat biscuits, 11 oatcakes, 5 infant cereals) were analysed for mycotoxins. Two oat porridge samples were tested for bioaccessibility using Infogest 2.0 method followed by faecal microbiota incubations. T-2/HT-2 and their glucosides were the most prevalent mycotoxins in unprocessed food oats (85–100%). DON was also highly prevalent (70%) while DON-glucoside, NIV, ZEN and their glucoside were less frequently detected. Reduction of 89–100% was achieved by de-hulling oats for most mycotoxins except DON-glucoside (77% reduction). Oat food samples were also frequently contaminated with T-2+HT-2 (prevalence porridge 90%, granola and muesli 40%, oat biscuits 36%, oatcakes 73%) while no mycotoxins were detectable in infant cereal foods. Upon in vitro digestion, 73–82% of free and glucosylated T-2/HT-2 were readily bioaccessible, underlining their importance for human dietary exposure. The study highlights that T-2/HT-2 contamination is prevalent in oats and carry-over into food products cannot be completely avoided, potentially leading to mycotoxin exposure in oat food consumers.

Toxins doi: 10.3390/toxins18070294

Authors: Bozidar Udovicki Dragan Milicevic Andreja Rajkovic

Risk assessment is a dynamic and continuously evolving process aimed at characterizing the potential adverse effects on life and health arising from exposure to hazards. Among these hazards, mycotoxins represent one of the most significant and widely investigated contaminants in food and feed. This review aims to summarize current knowledge on mycotoxin risk assessment practice, both for single substances and for cumulative risk assessment, and to provide practical guidance for future researchers. As individual substances, mycotoxins are well characterized from a toxicological perspective, and numerous risk assessments have been conducted globally across a wide range of food products. Although well-established methodological frameworks exist to support the cumulative risk assessment of mycotoxins, further efforts are needed to define common assessment groups, especially considering the pleiotropic nature of these compounds.