Toxins doi: 10.3390/toxins18100452

Authors: Josh A. Garber Damian Shea Juliette L. Smith

The co-occurrence of phycotoxins produced by harmful algal blooms is becoming more frequent. To combat this problem, monitoring programs that can track and quantify a wide range of hydrophilic and lipophilic phycotoxins are required. A 24 h, bench-scale study was conducted to provide an initial comparison of apparent sorption and extraction recovery for 24 freshwater and marine phycotoxins from two passive samplers under controlled artificial seawater conditions. Modifications were made to previously published ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methods to capture all phycotoxin analytes of interest in two injections. This analytical method was also evaluated alongside extraction methods for a commonly used passive sampler, a solid phase adsorption toxin tracker (SPATT), and a novel passive sampler, the composite integrated passive sampler (CIPS). SPATT contained a lipophilic resin, Diaion® HP20, whereas the CIPS is a continuous sheet with hydrophilic and lipophilic regions aimed at allowing for adsorption across a wider range of phycotoxins. A single-step extraction procedure was tested for both samplers to determine apparent extraction recoveries for all 24 phycotoxins. The CIPS seemed to produce overall higher apparent extraction recoveries of analytes, but the receiving phase generally sorbed less ng of phycotoxin during the experimental period than SPATTs and matrix effects were present. Across all lipophilic and hydrophilic phycotoxins, SPATT sorbed more than 90% of each phycotoxin from the seawater within 24 h, for all but four phycotoxins. The CIPS, however, sorbed less of each, but still more than 50% in artificial seawater for all but three phycotoxins. When considering extraction efficiency, SPATT may be better suited for hydrophilic phycotoxins, while the extraction method for CIPS appeared more efficient for lipophilic compounds, but further investigations are required to investigate potential matrix effects. The utility of the novel CIPS was preliminarily demonstrated in the laboratory, with seemingly relatively improved performance for particular phycotoxin classes (Azaspiracids, Dinophysistoxins/Pectenotoxins, Microcystins, and Brevetoxin) as compared to SPATT. The expanded UPLC methods and CIPS must be trialed in the field next to determine their applicability to monitoring, research, and management. The benchtop work, however, already suggests that users should consider the full range of phycotoxins present at an environmental site and create sampling schemes appropriate for their region given the different sorption and extraction efficiencies of these two samplers.

Toxins doi: 10.3390/toxins18100451

Authors: Avra Melina Madella Carlon Catharina Emma van van Gerven Elena Sendino Garví Jeroen van Bergenhenegouwen Johan Garssen Rosalinde Masereeuw Saskia Adriana Overbeek

Chronic kidney disease (CKD) is increasingly linked to intestinal barrier dysfunction, which contributes to systemic inflammation and disease progression. We investigated whether 2′-fucosyllactose (2′-FL), a human milk oligosaccharide, could modulate intestinal epithelial barrier function under uremic toxin exposure and examined the role of the aryl hydrocarbon receptor (AhR). Using wild-type (WT) and AhR knockout (AhR KO) Caco-2 cell lines, we assessed barrier function and cellular responses through transepithelial electrical resistance (TEER), paracellular permeability (Papp), tight junction (TJ) gene expression, AhR target gene activation, and metabolic assays. AhR deficiency impaired epithelial maturation, confirming its role in barrier development. Short-term exposure to uremic toxins (UTs) did not significantly affect TEER or paracellular permeability under the conditions tested. However, 2′-FL reduced paracellular permeability in both WT and AhR KO cells in the presence of FCS. These effects occurred without changes in TJ gene expression, indicating mechanisms beyond transcriptional regulation. AhR target genes were not induced by UTs, suggesting limited receptor activation, and metabolic activity was unaffected. Overall, 2′-FL reduced paracellular permeability under selected experimental conditions, while the impaired barrier development observed in AhR-deficient cells supports a role for AhR in intestinal epithelial barrier development.

Toxins doi: 10.3390/toxins18100448

Authors: Woon-Kyong Chung Min Joo Kim

More aesthetic patients now ask for changes in facial contour: a smaller face, a sharper jawline, and less sagging. Botulinum toxin (BoNT) is best known as a treatment for wrinkles, but it can also be used for modifying facial contour. With BoNT alone, a wide face can be made narrower, a heavy lower face can be made lighter, sagging on the jawline can be reduced, and a short chin can be corrected. Many physicians already do this, but each site is taught and studied on its own. This review brings the sites together under the name “contouring BoNT”. It describes seven sites: five muscles (masseter, temporalis, platysma, depressor anguli oris, mentalis) and two salivary glands (parotid, submandibular). The seven sites work in three ways: reducing the volume of a muscle or gland, removing a downward pull so that the face looks lifted, and relaxing an overactive muscle that changes the shape of the chin. All three serve one goal that patients describe in their own words: a smaller and lifted face. For each site, the review explains what it does to the contour, who is a good candidate, who needs caution, the published dose range and the authors’ practice, and which neighboring site changes when the target is treated. It explains which sites are checked together and how each is examined. Two safety points specific to contouring—the immunogenicity of large repeated doses, and salivary gland treatment—are discussed separately. The framework provided is a checklist for assessment, not a rule stating that every patient needs all seven sites to be treated.

Toxins doi: 10.3390/toxins18100449

Authors: Luyao Wang Li Ding Tianxiang Qin Xinyu Liu Chenglan Lou Hongyi Luo Fang Sun Chenhu Qin Xiangdong Ye Xudong Luo Zongyun Chen

Kunitz-type peptides from venomous and blood-sucking animals represent an important natural molecular resource for anticoagulant development towards XIa, a new target for antithrombotic drug discovery. However, the plasmin-inhibitory activity of these Kunitz-type XIa inhibitory anticoagulants limits their application. Here, using the snake Kunitz-type toxin analogue BF9-N17H, an XIa inhibitor with a unique P1-site histidine, as a template, P2′ scanning was performed to enhance its selectivity towards XIa. Ten BF9-N17H analogues were designed using different amino acid classification replacement at the P2′ site, including non-polar amino acids A and P, acidic amino acids D and E, polar amino acids S, N, and Y, and basic amino acids K, R, and H. Coagulation factor inhibitory experiments showed that three BF9-N17H analogues, BF9N17HP2, BF9N17HK8 and BF9N17HR9, lost their XIa inhibitory activity, but seven other analogues, BF9N17HA1, BF9N17HD2, BF9N17HS4, BF9N17HN5, BF9N17HY6, BF9N17HE7 and BF9N17HH10, effectively maintained their inhibitory activity towards XIa, with IC50 values from 61.19 ± 7.04 nM to 4.54 ± 3.65 nM. One analogue, BF9N17HD2, inhibited XIa with an IC50 of 6.19 ± 1.38 nM and had a selectivity index of 142.79 towards plasmin. It only had weak inhibitory activity toward plasmin, with an IC50 of 883.88 ± 148.35 nM. Excitingly, five analogues, BF9N17HS4, BF9N17HN5, BF9N17HY6, BF9N17HE7 and BF9N17HH10, almost lost their plasmin-inhibitory activity completely, indicating their high selectivity towards XIa. To the best of our knowledge, this is the first report of Kunitz-type XIa inhibitors without plasmin-inhibitory activity, and it highlights that the Kunitz-type peptide scaffold might be used as a potential candidate for the development of potent and selective XIa-inhibitory antithrombotic agents.

Toxins doi: 10.3390/toxins18100447

Authors: İsmet Bulut Zeynep Yegin Katran Özge Argın Mustafa Katran Fatma Çakmak

Venom immunotherapy (VIT) is the only disease-modifying treatment for Hymenoptera venom allergy, but real-world data describing VIT-related reactions and individualized premedication practices remain limited. We retrospectively evaluated 16 adults undergoing VIT with a target maintenance dose of 100 µg administered every 4 weeks at a tertiary allergy center between January 2024 and December 2025, focusing on clinical characteristics, premedication, treatment-related reactions, and field-sting outcomes during follow-up of up to 24 months. All patients had severe index reactions (Müller grade III-IV); 13 (81.3%) received Vespula VIT and 3 (18.8%) honey bee VIT. Premedication was initiated before VIT in seven patients (43.8%), including omalizumab in five (31.3%). During the 7-week up-dosing phase, five patients (31.3%) developed local reactions, whereas no systemic reactions occurred (0/16; 95% CI: 0–20.6%). During maintenance, one systemic reaction occurred (6.3%), requiring adrenaline and one-night intensive care admission, after which VIT was discontinued. Two patients were re-stung during follow-up and developed only local reactions. In this small, selected real-world cohort, no systemic reactions were observed during the up-dosing phase, whereas one systemic reaction occurred during maintenance. Given the limited sample size, wide confidence intervals, and frequent use of individualized premedication, these findings should be interpreted as descriptive observations and not as evidence establishing the safety of VIT or the effectiveness of premedication.

Toxins doi: 10.3390/toxins18100450

Authors: Rui Xie Bei Qin Naila Khan Bangash Mingyao Xie Kunlong Yang Fengqin Song

Pyruvate carboxylase (PYC), a multifunctional biotin-dependent enzyme, is essential in diverse metabolic cascades including gluconeogenesis and the tricarboxylic acid (TCA) cycle. This study aimed to dissect the physiological roles of PYC and its succinylated site in Aspergillus flavus (A. flavus). In A. flavus, biological functions of PYC were characterized through gene deletion and site-directed mutagenesis. Phenotypic analysis and qRT-PCR demonstrated that PYC positively regulated sporulation and sclerotial formation, while negatively regulating aflatoxin B1 biosynthesis. The transcriptomics revealed that development-related DEGs of abaA, wetA, vosA, atfB, creA, crzA, and areA were downregulated, whereas veA and are2 were increased in Δpyc1. Meanwhile, aflatoxin-related DEGs of nor1, aflW, agiB, aflO, and aflM were upregulated, while cfoC and ucdC were lower in Δpyc1. In addition, a succinylated site on PYC (K1068) was characterized by noncovalent interactions. Combined with the transcriptomics analysis of this site, this study revealed that DEGs in pyc1K1068R (mimic desuccinylation) were enriched in fundamental metabolic pathways of glycolysis/gluconeogenesis, carbon metabolism, fructose and mannose metabolism, pentose phosphate pathway, etc., which mainly affected A. flavus development. This research provided insights into the regulatory mechanisms underlying A. flavus development and mycotoxin contamination.

Toxins doi: 10.3390/toxins18100446

Authors: Dario Lucchetti Daniela Delfino Marta Senofonte Laura Spinaci Rocco Baccelliere Alessandro Ubaldi Francesca Longo

The presence and accumulation of marine biotoxins, including paralytic shellfish toxins (PSTs) and tetrodotoxin (TTX) analogues, in marine organisms poses a significant food safety concern. This study describes the validation of a quantitative confirmatory HILIC–LC–MS/MS method for the simultaneous determination of a broad panel of marine toxins in four matrices: mussels, clams, sea snails, and sea urchins. The method covered 17 target compounds, including TTX analogues (TTX, 4-epiTTX, and 4.9-anhTTX), saxitoxin and analogues (STX, dcSTX, NEO, and dcNEO), gonyautoxins (GTX 1–4, GTX 2–3, GTX 5, GTX 6, and dcGTX 2–3), and sulfocarbamoyl toxins C1–C2. Particular attention was devoted to the evaluation of matrix effects (ME), a critical factor affecting LC–MS/MS quantification. The method was validated according to SANTE/11312/2021 guidelines, showing satisfactory selectivity, precision (RSD ≤ 20%), and accuracy (recoveries within 70–120%). Significant matrix-dependent signal suppression and enhancement were observed among the investigated species. Matrix-matched calibration enabled reliable quantification in mussels, clams, and sea snails, whereas sea urchin samples showed greater analytical complexity. The validated method provides a robust approach for the determination of PSTs and TTX analogues in different marine organisms and supports its application in official monitoring programs.

Toxins doi: 10.3390/toxins18100445

Authors: Yo Han Lee Ye Rin Bae Tae Hong Kang Jung Woong Yoon Seung Joon Lim Chae Hyun Lee Suhyeon Yun Sang In Lee

Fumonisin B1 (FB1) is a mycotoxin commonly found in animal feed, particularly in corn-based products. Exposure to FB1 can damage multiple organs, including the intestine, but its molecular effects on intestinal epithelial cells remain incompletely understood. This study investigated FB1-induced toxicity in porcine intestinal epithelial (IPEC-J2) cells and screened natural products for protective effects. Exploratory gene expression profiling identified genes showing predefined expression changes following FB1 exposure, and subsequent Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated associations with apoptosis-related processes. Annexin V/propidium iodide staining showed increased apoptosis-associated staining, while quantitative real-time polymerase chain reaction (qRT-PCR) showed altered expression of BCL2, CASP3, and CASP9. To identify protective candidates, 332 natural products were evaluated using high-throughput screening. Among the screened extracts, C. communis showed the highest cell viability in the primary screening and was selected for further investigation. Co-treatment with C. communis was associated with reduced apoptosis-associated staining and partial counteraction of FB1-associated changes in apoptosis-related gene expression. These findings suggest that C. communis warrants further investigation as a natural product for mitigating FB1-associated intestinal epithelial toxicity.

Toxins doi: 10.3390/toxins18100444

Authors: Kyu-Ho Yi Kyu-Lim Lee

Botulinum neurotoxin type A (BoNT-A) dosing in facial aesthetics varies across anatomical targets, formulations, and treatment indications. This structured narrative review examined published dose ranges and the potential relevance of muscle thickness, target depth, functional anatomy, and injection localization to individualized treatment planning. English-language regulatory, clinical, anatomical, ultrasound, and consensus literature available through 13 April 2026 was considered. Evidence was interpreted according to source type, distinguishing approved product-specific regimens, controlled clinical studies, observational reports, and anatomy- or expert-derived recommendations. Muscle bulk may inform masseter contouring, but the literature does not establish a validated thickness-based dosing equation applicable across facial muscles. For facial expression muscles, contraction patterns, treatment objectives, and preservation of adjacent function are central considerations. Evidence for aesthetic dosing of the temporalis and salivary glands remains limited, and glandular indications require separate assessment from skeletal muscle treatment. Anatomically informed placement has a plausible mechanistic rationale, whereas evidence that motor-zone targeting consistently reduces toxin requirements is insufficient. Ultrasound may assist selected anatomically complex or previously treated cases but is not established as necessary for routine injections. Safe dose selection requires formulation-specific interpretation, indication-specific evidence, individualized anatomical assessment, and recognition of the limitations of existing studies.

Toxins doi: 10.3390/toxins18100443

Authors: João G. Brás Fokko P. Wieringa Babette H. Lentferink Simon Kootwijk Albert H. Westra Leonard J. van Schelven Mies van Steenbergen Joachim Jankowski Marianne C. Verhaar Karin G. F. Gerritsen Jeroen C. Vollenbroek

Current hemodialysis (HD) methods inefficiently remove protein-bound uremic toxins (PBUTs) due to their high binding affinity to plasma proteins, mainly albumin. This study aimed to investigate electromagnetic (EM) fields and temperature as potential extracorporeal, non-systemic methods to increase HD PBUT clearance. PBUT release was measured in infusible (caprylate-stabilized) albumin solutions spiked with indoxyl sulfate (IS) using an in vitro dialysis setup including a real-time, in-line IS-sensor. No measurable influence on PBUT release was observed during EM field-tests (f = 1–600 MHz, E <= 1 kV/m), but temperature tests (35–40 °C range) showed an absolute increase in IS free fraction of 0.7%/°C. Furthermore, static equilibrium dialysis tests, including IS, p-cresyl sulfate (pCS) and hippuric acid (HA), showed a free fraction increase for all three PBUTs upon a temperature increase from 37 °C to 43 °C (IS 1.30-fold, pCS 1.12-fold, HA 1.15-fold). Mathematical modeling showed a 1.14-fold decrease in plasma IS concentration after 5 weeks of HD, and exploratory hemocompatibility tests did not reveal pronounced changes over 15 heating cycles at 43 °C under static conditions. Extracorporeal temperature elevation is a promising technique to enhance PBUT removal during hemodialysis, which may be easily combined with current HD machines, does not break sterility, and avoids elevating free PBUTs inside the body.

Toxins doi: 10.3390/toxins18100442

Authors: Andrew D. Turner Karl J. Dean Monika Dhanji-Rapkova Alison O’Neill Benjamin H. Maskrey Patrycja Stubbs David M. Hartnell Shamina Page Naomi Baker Nadine Morrell John M. Turner Joan Turner Florence R. I. Turner Stephen D. Gregory Adam M. Lewis Katy Lewis

Cyanobacteria are microorganisms that can produce natural toxins with significant impacts on human, animal and ecosystem health. Portishead Marine Lake is a small freshwater lake in southwest England frequented by thousands of people each year for recreational activities but was associated periodically with prolific cyanobacterial blooms with potential risk to visitor health. Water samples were tested weekly between January and December 2016 for cyanobacteria and cyanotoxins. Citizen scientists undertook testing for nutrients, dissolved oxygen, pH, temperature and meteorological parameters. Four major blooms of Microcystis spp. were observed in the warmer months between May and September and were associated with high concentrations of microcystins. Complex toxin congener profiles were dominated by MC-LR and MC-RR, with lower contributions from MC-WR and MC-YR. Cyanobacterial cell densities and summed microcystin concentrations ranged from 1600 to 5.6 × 106 cells/mL and 0.1 to 18,069 µg/L, respectively, and correlated significantly. Toxin concentrations were positively correlated with nitrate levels and inversely correlated with pH, although mean pH levels did not fall below 7.0. There was little statistical evidence for influence of weather conditions overall, although significant intra-lake differences in cell densities and toxin concentrations were noted. Intervention measures employed by local authorities included the use of barley straw, hydrological manipulation, debris removal and ultrasound treatment. Whilst no statistical assessment of intervention effectiveness was possible without effective control samples, there was some limited evidence for the temporary reduction in toxin levels after hydrological intervention, but none following installation of ultrasound. Overall, the study utilized LC-MS/MS and microscopy to quantify microcystins and Microcystis spp., respectively, and evidenced risks to lake visitors and the importance of toxin testing in recreational waters and the use of appropriate risk mitigation approaches. Further work incorporating appropriate experimental controls would be needed to determine the effectiveness of hazard mitigation measures for reducing or eliminating human health threats.

Toxins doi: 10.3390/toxins18100441

Authors: Veronica Russo Sara Ragucci Miriam Russo Maria Antonietta Oliva Nicola Landi Antimo Di Maro Antonietta Arcella

Background: Glioblastoma (GBM) is the most aggressive primary brain tumor and remains difficult to treat because of its highly invasive behavior and resistance to conventional therapies despite surgery, radiotherapy, and chemotherapy. Sodin 5, a novel type-1 ribosome-inactivating protein (RIP), inhibits protein synthesis and displays cytotoxic activity against several tumor cell types, making it a promising candidate for GBM therapy. Methods: The antitumor activity of sodin 5 was evaluated in the U87MG glioblastoma cell line and patient-derived primary GBM cell lines (CG, ZAR, and NULU). Cell viability, migration, and molecular changes were assessed after treatment. Biological activity was also investigated under different pH conditions. Vimentin and phosphorylated ERK (p-ERK) expression were analyzed to investigate the mechanisms underlying sodin 5 activity. Results: Sodin 5 significantly reduced cell viability in all GBM cell lines. Treatment also inhibited cell migration, accompanied by decreased vimentin expression, suggesting cytoskeletal remodeling and reduced invasive potential. Furthermore, sodin 5 decreased p-ERK levels, indicating modulation of signaling pathways involved in proliferation and motility. The protein retained biological activity under different pH conditions, supporting its potential efficacy in the heterogeneous GBM microenvironment. Conclusions: These findings identify sodin 5 as a promising therapeutic candidate for GBM by targeting multiple processes involved in tumor growth and invasion.

Toxins doi: 10.3390/toxins18100440

Authors: Stephany Young Yusty Celestino Aguilar Alexa Prescilla-Ledezma Alexander A. Martínez Claudia González Hildaura Acosta de Patiño

Snakebite envenoming is a major neglected public health problem in Panama, yet culture-independent data on the oral microbiota of medically important viperid snakes remain limited. We characterized the oral microbiota of four captive species native to Panama (Bothrops asper, Bothriechis nigroviridis, Cerrophidion sasai, and Porthidium lansbergii) using forward-read analysis of 16S rRNA V3–V4 amplicons and compositional methods. Among 68 retained oral samples, communities were dominated by Pseudomonadota, Bacillota, Actinomycetota, and Bacteroidota. Observed ASV richness differed among species (Kruskal–Wallis H = 21.87, p < 0.001), whereas no significant differences were detected in Shannon diversity or Pielou’s evenness. Richness differences require caution because sequenced negative controls were unavailable and no independent depth-aware sensitivity analysis was performed. Community composition was significantly associated with host species (PERMANOVA R2 = 0.197, p = 0.001), although PERMDISP detected heterogeneous within-group dispersion (p = 0.005), requiring cautious interpretation. ANCOM-BC2 identified 21 differentially abundant genera, including Providencia, Morganella, Pseudomonas, Stenotrophomonas, and Staphylococcus, with high relative abundance of Mycoplasma in a subset of P. lansbergii samples. These findings provide a culture-independent taxonomic baseline for future culture-based, antimicrobial-susceptibility, and paired snake-oral/human-wound investigations in Panama.

Toxins doi: 10.3390/toxins18100439

Authors: Xiaoyu Xiao Junxuan Zeng Songheng Zheng Shifeng Liu Qi Deng Mei Qiu Zhijia Fang Gaoshuang Hu Yaling Wang Youxin Wang

Microcystins (MCs) produced by cyanobacteria can contaminate aquatic foods, but drinking water surveillance does not directly characterize dietary exposure. This critical narrative and framework review evaluates environmental transfer, tissue residues, analytical coverage, toxicity, exposure assessment, and management for fish, bivalves, crustaceans, and related products, with a separate assessment of algal dietary supplements. Evidence is organized by species, consumed tissue, concentration basis, analytical fraction, congener coverage, and consumption scenario. Reported concentrations are not directly comparable when studies analyze different organs or use different extraction procedures and mass bases. Paired analyses of tilapia muscle illustrate how measurements limited to free congeners can miss a substantial additional burden, while shellfish studies show why clearance requires verification in the edible tissue. Structural and toxicological evidence explains why MC-LR provides a reference for assessment but does not fully characterize every mixture. We propose a framework linking environmental warnings to product sampling, validated quantification, explicit exposure scenarios, and proportionate management. Priorities are comparable occurrence reporting, interlaboratory validation, characterization of the oral relevance of bound fractions, and congener-specific toxicity data. Neither a positive screening result nor an exceedance of a chronic intake benchmark alone establishes the probability of illness from an individual meal.

Toxins doi: 10.3390/toxins18100438

Authors: Ling Wang

Mycotoxins, toxic secondary metabolites produced by filamentous fungi, represent a persistent challenge to global food safety, animal health, and human well-being [...]

Toxins doi: 10.3390/toxins18100437

Authors: Christopher Hernandez Vincent Merlhiot Selma P. Snini Florence Mathieu Vanessa Durrieu Jean-Denis Bailly

Aflatoxins are carcinogenic mycotoxins that can contaminate many crops and whose distribution is increasing due to climate change. Given the mutagenic and carcinogenic properties of aflatoxins, protecting public health requires minimizing dietary exposure. In the European Union, maximum regulatory levels can be as low as 0.10 µg/kg for AFB1 in processed cereal-based foods and baby foods. Once produced, aflatoxins are stable, so the best way to protect consumers is to minimize aflatoxin production. Numerous studies have investigated sustainable and eco-friendly strategies to limit aflatoxin contamination in food and feed. These strategies rely on microorganisms or natural, plant-derived compounds that hinder the development of aflatoxigenic species and/or specifically inhibit toxin biosynthesis. However, a substantial translational gap remains between initial in vitro evidence of the effects on fungal growth and/or aflatoxin synthesis and the effective application of such biocontrol agents under field conditions. This review aims to clarify the current state of the art by presenting the diverse biocontrol strategies currently being investigated to combat aflatoxins. We discuss the advantages and disadvantages of such strategies and describe the level of technological readiness, which spans from in vitro tests to large-scale in-field applications.

Toxins doi: 10.3390/toxins18100436

Authors: Tamás Schieszl György Kövér Lucia Gambacorta Michele Solfrizzo Lajos Biró Györgyi Arató Andrea Zentai Brigitta Bóta András Szabó Melinda Kovács

Mycotoxins are common contaminants of the food and feed chain and have been associated with developmental disorders, hepatotoxicity, nephrotoxicity, and various types of cancer. This study aimed to estimate dietary mycotoxin exposure in the Hungarian population and to identify the major cereal-based food contributors. Urinary mycotoxin concentrations were measured by LC–MS/MS, and population exposure was assessed using estimated daily intake (EDI) values. Associations between food consumption and urinary mycotoxin levels were evaluated using Spearman correlation analysis on urine samples collected in year 2018 and 2020. The average EDI values were below the tolerable daily intake (TDI) for all mycotoxins in both years, except for ochratoxin A in 2018. Distinct exposure patterns were observed between the two sampling years. In 2018, consumption of oats and oat-based products showed a moderate positive correlation with urinary fumonisin B2 levels (r(s) = 0.497, adjusted p = 0.001962350574942). In 2020, wheat and wheat-based product consumption correlated with urinary deoxynivalenol levels (r(s) = 0.6868, adjusted p = 0.009509496), while corn and corn-based product consumption correlated with urinary zearalenone levels (r(s) = 0.6137, adjusted p = 0.04134226737). These findings highlight differences in dietary mycotoxin exposure between the studied populations and sampling periods and emphasize the importance of human biomonitoring to identify major dietary mycotoxin sources/carriers and support exposure and risk assessment.

Toxins doi: 10.3390/toxins18100435

Authors: David M. Simpson Marta Banach Jason Aldred Jack Lee William Ondo Fernando Pagan Anastasia Bagretsova Gudrun Klein Thorin L. Geister Irena Pulte Mandar Jog

This double-blind, placebo-controlled, phase II trial (NCT04766723) investigated the efficacy and safety of incobotulinumtoxinA (Xeomin®) in the treatment of essential tremor (ET) of the upper limb (UL). Adults with ET of the UL were randomized 2:1 (n = 78) and treated (n = 77) with intramuscular injections of incobotulinumtoxinA or placebo at semi-flexible doses into their motor-dominant arm. The primary endpoint was the change from baseline to week 6 in maximum tremor amplitude at wrist level. Baseline characteristics of treated patients were well matched across treatment arms. The primary endpoint was not met; maximum tremor amplitude at the wrist decreased from baseline to week 6 in both groups, with changes numerically favoring placebo (least squares mean treatment difference 0.21, p = 0.28). At week 6, compared with the placebo group, the incobotulinumtoxinA group had improvement in The Essential Tremor Rating Assessment Scale (TETRAS) performance dominant UL score (p = 0.006) and Quality of Life in ET (QUEST) total score (p = 0.073). However, these secondary outcomes must be interpreted descriptively as exploratory findings. Although the p-value and the 95% confidence interval including zero did not support statistical significance, the observed numerical treatment difference in change from baseline in QUEST total score suggests that the treatment effect may be consistent with the minimal clinically important difference in favor of incobotulinumtoxinA. Improvement in TETRAS Activities of Daily Living UL score was similar in both groups at week 6 (p = 0.933). IncobotulinumtoxinA was generally well tolerated. Treatment of UL ET with incobotulinumtoxinA did not show superiority over placebo for the primary endpoint. Secondary and other efficacy endpoints should therefore be viewed as exploratory, generating hypotheses suggesting potential improvements in TETRAS UL performance score and quality of life that require confirmation in further research. IncobotulinumtoxinA demonstrated a favorable safety profile without any new or unexpected safety concerns.

Toxins doi: 10.3390/toxins18100434

Authors: Hiromi Watari Koji Nakano Masaya Kitano Hiromu Kageyama Yoshikazu Tanaka Yuri Ishibashi Ryuichi Sakai

Soritesidine (SOR) is a potent proteinaceous toxin from the marine sponge Spongosorites sp. that is toxic to mammalian cells, brine shrimp larvae, sea hare embryos, and mice. Although SOR exhibits divalent metal ion-dependent DNase activity, its molecular identity and mechanism of action remained unknown. We determined the amino acid sequence of SOR and investigated its cellular mode of action. Degenerate PCR, 3′-RACE, inverse PCR, and nested PCR identified an open reading frame encoding a 947-amino-acid acidic protein. The deduced sequence showed no substantial similarity to known proteins except for approximately 30% sequence identity to a hypothetical protein from the marine α-proteobacterium Euryhalocaulis caribicus. Analysis of the AlphaFold 3-predicted N-terminal domain using Foldseek and structural superposition revealed a DNase I-like fold with a catalytic architecture similar to that of vertebrate DNase I. A recombinant N-terminal fragment (residues 1–310, N-SOR) exhibited DNA cleavage activity, supporting the assignment of this region as a DNase I-like endonuclease domain. The recombinant fragment lacked detectable cytotoxicity, suggesting that regions outside the N-terminal DNase I-like domain contribute to the cytotoxicity of SOR. Fluorescence imaging showed that SOR entered cells and accumulated in the nucleus, whereas its cellular uptake was attenuated at 4 °C, suggesting the involvement of an energy-dependent internalization pathway. Immunostaining revealed increased γH2AX signals in the nuclei of SOR-treated cells, indicating activation of a DNA damage response. Together, these findings identify SOR as an unusual multidomain protein toxin containing a functional DNase I-like domain and provide a molecular basis for further investigation of its cytotoxic mechanism.

Toxins doi: 10.3390/toxins18100433

Authors: Yucong Li Xiang Wan Fangqun Gan

Microcystin (MC) risk is often assessed within bloom-affected lakes and reservoirs, although hydrological connectivity can extend exposure into downstream river and estuarine networks. We synthesize evidence using a lake–river continuum framework that treats connected reaches as active processing environments. The framework separates the source pool, export flux, and downstream exposure, and distinguishes advective redistribution, reversible retention, irreversible transformation, and source or remobilization processes. Hydrology sets reaction opportunity, while phase partitioning, photochemical and microbial transformation, sediment exchange, and biological uptake determine how MC mass and bioavailability change during transport. Field evidence shows that downstream occurrence cannot be inferred reliably from visible source blooms: reaches may attenuate, retain, redistribute, or locally renew MC exposure depending on hydraulic state and process coupling. These dynamics shift exposure among water, particles, sediments, cells, and biota. Consequently, source-lake indicators or water concentrations alone can mischaracterize ecological exposure, and drinking-water guideline values are not ecological protection thresholds. Key priorities are flux-resolved monitoring, direct field estimates of process rates and transformation products, chronic ecotoxicology matched to realistic exposure routes, and hierarchical transport–biogeochemical models. This continuum perspective links source, process, pathway, receptor, and effect within a single source-to-risk framework.

Toxins doi: 10.3390/toxins18100432

Authors: Safy M. Sh. M. Abaza Jiaqi Zhou Ziqin Wang Zanyu Pan Haoran Ma Yingdi Sun Ruben K. Dagda Edward S. Gasanoff

Bee venom melittin is a small protein with significant pharmacological potential. In this study, AutoDock simulations identified Arg22 as a key residue of melittin responsible for targeting the phosphate groups of cardiolipin (CL). Recombinant melittin with the native amino acid sequence, as well as a modified version with Arg22 substituted by Gly22, were cloned, expressed, and purified. The effects of both melittins on proton adsorption in CL-containing membranes, mitochondrial ATP production, and the formation of non-bilayer lipid structures were examined. Based on the array of computational and experimental data obtained in this work, together with published findings, we propose that Arg22, in tandem with Lys23, targets and neutralizes CL bound to conserved Lys7 and Lys43 of the ATP synthase rotor, thereby detaching CL from the rotor. These events promote a conical shape of CL, which triggers the formation of inverted micelles that entrap protons from the Fo subunit and transfer them to the proton pump of a respiratory complex. This study advances both the conceptual framework of mitochondrial energetics and the translational application of natural toxins. Specifically, the findings support the development of new pharmaceuticals aimed at counteracting the loss of mitochondrial energy production that occurs with aging and pathological conditions.

Toxins doi: 10.3390/toxins18100431

Authors: Yuval Levy Ayalon Reis Galit Blecher Oksana Daron Sara Dadon Raz Zarivach Noam Zilberberg

Kv4 channels generate transient A-type potassium currents, but the contribution of Kv4.1 remains difficult to define because few pharmacological tools discriminate among Kv4 isoforms. We investigated the molecular basis of Kv4.1-selective inhibition by BlTx2, an α-KTx15 scorpion toxin from Buthacus leptochelys. Using electrophysiology, mutagenesis, tandem-channel analysis, structural modeling, and evolutionary analysis, we mapped the determinants governing BlTx2 activity and selectivity. Substitution of conserved α-KTx residues Lys27 and Tyr36 abolished detectable inhibition, whereas N30A reduced potency, supporting a canonical pore-blocking mechanism. A Kv4.1/Kv4.3 tandem chimera reduced BlTx2 potency, consistent with a contribution of adjacent Kv4.1 subunits to the high-affinity toxin-binding surface. Channel mutagenesis identified the Kv4.2/Kv4.3 lysine corresponding to Kv4.1 Asn355 as a key barrier to BlTx2 activity, since introducing this lysine into Kv4.1 abolished detectable inhibition. Mutational mapping of BlTx2 revealed that pore-facing residues required for inhibition are separated from peripheral residues that tune potency and cross-reactivity. In particular, Q9K and I18K conferred Kv1.2 activity, partly through interaction with Kv1.2 Glu355. Evolutionary analysis linked conserved pore-facing residues to purifying selection, and peripheral determinants to diversification. These findings identify distinct conserved and peripheral determinants that together shape BlTx2 activity and selectivity, thereby establishing BlTx2 as a useful probe for Kv4.1 function.

Toxins doi: 10.3390/toxins18100430

Authors: Tomé Azevedo Lucas Ribeiro Joana Azevedo Rui Barbosa Rita Mendes Joana Martins Marisa Freitas Vitor Vasconcelos Alexandre Campos

Cyanobacterial blooms and the associated production of cyanotoxins represent a growing concern for freshwater ecosystems, particularly in regions increasingly affected by eutrophication and climate change. In Portugal, data on the occurrence and distribution of cyanotoxins remain limited, especially in southern multipurpose reservoirs that support the water supply and irrigation in a region severely affected by water scarcity. This study investigates the presence of cyanobacteria and their major cyanotoxin groups—microcystins (MCs), cylindrospermopsin (CYN), and anatoxin-a (ANTX)—in three reservoirs of the Alqueva water storage system (EFEMA), combining chemical analyses with molecular tools to assess toxin-producing cyanobacteria. Liquid chromatography–mass spectrometry (LC–MS) detected MC-LR, MC-RR, and MC-YR exclusively in the particulate fraction of water samples, while no toxins were detected in the dissolved fraction. MC concentrations were generally low (<0.1 µg/L), with the highest levels observed during summer, particularly in July. Among the variants, MC-LR was the most common and abundant. Molecular analyses (16S rRNA) confirmed the widespread presence of cyanobacteria and identified the genus Microcystis in all reservoirs. Several genes from the microcystin biosynthesis cluster (mcyA, mcyB, mcyD, mcyE, and mcyG) were amplified, and sequencing of the mcyA gene revealed high homology with Microcystis aeruginosa. Although the detected MC concentrations remained below World Health Organization (WHO) guideline values for drinking water, their recurrent detection and the confirmed presence of toxin-producing genotypes highlight the need for continued monitoring. This study underscores the value of integrating chemical and molecular approaches for assessing cyanotoxin risks in large multipurpose reservoir systems under increasing environmental pressure.

Toxins doi: 10.3390/toxins18100429

Authors: Stephen A. Ashford Mark Weatherall Klemens Fheodoroff Jorge Jacinto Richard J. Siegert Christian Hannes Lynne Turner-Stokes

Improvement in active function (making it easier to use the limb) is a common goal following treatment of upper limb spasticity with botulinum toxin (BoNT-A). ULIS-III was a large international, observational, longitudinal study (N = 953) conducted in the context of usual clinical practice over two years. In this secondary post hoc analysis, we examine whether goals for improvement in active function were met over repeated injection cycles. Patients were selected if improved active function (AF) was a primary/secondary goal for at least one cycle of treatment (n = 286/953). They were assessed at the start and end of each cycle using the Goal Attainment T-score alongside the Arm Activity measure (ArmA). Serial changes and predictors of improvement were explored. Across eight cycles, there were 1167 injections for active function, 875/1998 (44%) AF goals were set and 493 (57%) of those goals were achieved. Patients continued to show significant improvement in AF (p < 0.001) for each injection at least up to three cycles, which is likely to be clinically significant, especially in patients with milder spasticity who were naïve to BoNT-A. No statistically significant predictors were confirmed on mixed effects modeling. Conclusions: Repeated BoNT-A continued to produce significant improvement in active function for at least three cycles, regardless of patient- or treatment-related factors in this cohort, but type II errors due to small numbers cannot be excluded.

Toxins doi: 10.3390/toxins18100428

Authors: Kazuya Yoshida

Botulinum neurotoxin (BoNT) therapy has expanded from focal chemodenervation to a versatile therapeutic modality used across multiple medical specialties. The stomatognathic system is particularly suited to BoNT therapy because motor, sensory, articular, secretory, and aesthetic functions converge within a compact anatomical region. Current and emerging applications include oromandibular dystonia, dyskinesia, bruxism, tremor, hemimasticatory spasm, embouchure dystonia, masticatory muscle and temporomandibular disorder-related pain, trigeminal neuralgia, recurrent temporomandibular joint dislocation, sialorrhea, Frey syndrome, masseter prominence, gummy smile, and perioral wrinkles. Evidence and regulatory status vary substantially among indications, and many applications remain off-label. Dentistry and oral and maxillofacial surgery have historically played a limited role in BoNT therapy despite their detailed anatomical and functional expertise in this region. However, interest is increasing, and the recent approval of onabotulinumtoxinA in Japan for masseter prominence in adults younger than 65 years represents an important milestone. This review synthesizes current applications, limitations, safety considerations, and future directions, and proposes the stomatognathic system as a major expanding frontier for BoNT therapy.

Toxins doi: 10.3390/toxins18100427

Authors: Christine Rasetti-Escargueil Stefano Palea

The present review is focused on a range of BoNT/A molecular targets and the subsequent modulation of cellular signaling pathways focused on gene regulation, skin and nerve regeneration, vascular effects, and contributions to pain relief. This brings fresh insights to the diversity of BoNT/A mechanisms of action, opening new avenues of research into therapeutic options for patients. A summary of the mechanisms of action of BoNT/A formulations will be presented in parallel with their specific indications, showing the versatility of the BoNTs’ efficiency through different pathways well beyond the canonical SNARE cleavage mechanism. The current research progress is summarized, ranging from the molecular mechanism of BoNT/A involved in the treatment of inflammation, pathological scars, pain, and a range of neurological and vascular disorders via clinical investigations and experimental studies.

Toxins doi: 10.3390/toxins18100426

Authors: Imene Ghorbal Sara Redouane-Salah Safa Smail Amandine Nachtergael Mohammed Aufy Mohamed F. Abdallah

Mycotoxigenic fungi, particularly species of Aspergillus, Fusarium, and Penicillium, contribute substantially to food contamination through the production of chemically diverse mycotoxins. Plant-derived essential oils (EOs) and extracts have therefore attracted increasing interest as potential alternatives or complements to conventional antifungal strategies. The genus Artemisia, which comprises several aromatic species with extensive biological properties, may constitute a promising source of such natural agents. This review critically examines the available data on the use of EOs and extracts from four Artemisia species (A. absinthium, A. campestris, A. herba-alba, and A. judaica) against mycotoxigenic fungi, with an emphasis on chemical composition, antifungal efficacy, mechanisms of action, and safety. Across the available studies, evidence suggests that Artemisia preparations frequently inhibit fungal growth, but their efficacy varies substantially according to species, geographical origin, plant part, harvesting conditions, extraction method, chemical composition, fungal species, and experimental assay. Importantly, evidence for direct inhibition of mycotoxin biosynthesis remains considerably more limited than evidence for fungal growth inhibition. Proposed antifungal mechanisms include disruption of the fungal cell wall and plasma membrane, altered mitochondrial function, and interference with cellular transport processes, although mechanistic evidence specifically for Artemisia preparations remains limited. Available toxicological data further indicate that efficacy and safety must be evaluated together, particularly for chemotypes containing potentially hazardous constituents such as thujone. Overall, Artemisia EOs and extracts may represent promising but not yet sufficiently validated candidates for food-associated mycotoxin control. Future studies could prioritize chemical standardization, mechanistic characterization, mycotoxin-specific endpoints, food-matrix validation, and integrated efficacy–safety assessment.

Toxins doi: 10.3390/toxins18100425

Authors: Tanno Van Herpen Radja Millenio Kosasih Jeroen Kool Julien Slagboom

Between 80,000 and 140,000 fatalities occur yearly because of snakebite envenoming, which causes it to be classified as a Neglected Tropical Disease (NTD), stressing the importance of snake venom toxin characterisation to aid the development of next-generation antivenoms. Snake venom is made up of a mixture of bioactive proteins that are also known as venom toxins. These venom toxins, being complex and structurally diverse, present a significant challenge for analytical separation and functional characterisation. Traditional methods, such as reversed-phase liquid chromatography (RPLC), often involve denaturing conditions that can alter the native state of these proteins, potentially reducing their bioactivity and limiting subsequent bioassays. Therefore, this study developed and optimised an analytical methodology using non-denaturing Hydrophobic Interaction Chromatography (HIC) for venom separation in conjunction with post-column nanofractionation to perform post-column toxin identification using high-throughput venomics with parallel coagulation bioassays. The results demonstrate that the combination of HIC and nanofractionation allows separation and qualitative profiling of venom toxins with detectable post-column activity, although several toxins co-elute. The post-column bioassays demonstrated detectable coagulation-modulating activity, and high-throughput venomics provided rapid toxin identification capabilities and qualitative information on candidate venom proteins. This approach offers a robust platform for the detailed study of venom toxins, facilitating the discovery of novel bioactive compounds and enhancing our understanding of venom biology.

Toxins doi: 10.3390/toxins18100424

Authors: Yue Lin Yiseul Choi Xiaoqing Liu Hee-Dae Jeon Hyun Jin Park

Regulatory approval may occur before or after a research topic becomes prominent in the scientific literature. We examined this temporal relationship using botulinum neurotoxin (BoNT) as a longitudinal case study. We analyzed 2065 anatomy-related BoNT publications indexed in Web of Science and Scopus from 1983 to 2024. The literature was divided into 16 periods based on U.S. FDA approval dates for onabotulinumtoxinA, and indication-related keyword rankings and thematic clusters were compared with the corresponding approval dates. Among the 10 indication groups examined, 7 showed prominent indication-related keywords before FDA approval, 2 showed signals around the time of approval, and 1 showed a post-approval signal. We describe these patterns as “proactive exploration” when scientific attention became prominent before approval and “reactive catalysis” when it became more prominent around or after approval. Differences in the observed lag may be related to target anatomical complexity and market orientation, but this remains conceptual. Overall, the findings show that time-resolved scientometrics can retrospectively identify scientific patterns associated with regulatory milestones. The approach is hypothesis-generating and requires prospective validation before it can be used for regulatory forecasting.

Toxins doi: 10.3390/toxins18100423

Authors: Kathleen D. Cusick Jackie Palau Malcolm N. McFarland Edith A. Widder

Pyrodinium bahamense is a saxitoxin-producing bioluminescent dinoflagellate that forms annual recurring blooms in the Indian River Lagoon (IRL), FL. Little information exists as to the extent of bloom toxicity. This study examined toxin dynamics in P. bahamense IRL field populations from two sites in the northern IRL (NIRL). Additionally, genomic diversity and transcription were assessed using quantitative molecular assays targeting key functional genes in P. bahamense ecology: sxtA4 (toxin biosynthesis), rbcL (photosynthesis), and lcf (bioluminescence). A positive correlation was found between sxtA4 transcript copy number and toxicity, mirroring the trend recorded in previous seasons and indicating that sxtA4 transcripts can serve as a molecular proxy for bloom toxicity. Lab experiments with 32 strains isolated from the NIRL showed a large variation in copy number for both sxtA4 (42–318 copies cell−1, dependent on strain) and rbcL (1385 to 4911 copies cell−1). An lcf qPCR assay developed here showed substantial variation among isolates as well. These results are indicative of the underlying diversity in field populations, as genome copy number variation is a major driver of genome evolution and diversity. A negative correlation was found between cell abundance and toxin quota in field populations. Growth and toxin quota were also found to have a negative correlation in the 32 isolates. Based on these collective lab and field results, a model is presented in which P. bahamense populations utilize both their toxicity and bioluminescence in bloom development and persistence.

Toxins doi: 10.3390/toxins18100422

Authors: Suguru Yamamoto Takahiro Tanaka Nobutaka Kitamura Miho Kawachi Akio Yokoseki Minako Wakasugi Osamu Onodera Naoto Endo Masakazu Wada Kenji Sato Kaori Kikuchi Ayano Konagai Shin Goto Ichiei Narita

Cognitive dysfunction is increasingly recognized as a significant complication of chronic kidney disease (CKD), but the clinical relevance of protein-bound uremic toxins (PBUTs) to cognition remains insufficiently characterized. This cross-sectional analysis included 1217 community-dwelling individuals. Cognitive function was assessed using the Mini-Mental State Examination (MMSE), with scores < 28 indicating possible mild cognitive impairment (MCI). Serum concentrations of five PBUTs—indoxyl sulfate, p-cresyl sulfate, phenyl sulfate, indoleacetic acid, and hippuric acid—were measured, and a composite PBUT score was derived using principal component analysis. Multivariable logistic regression was used to examine the associations of MCI with the PBUT score and with each PBUT. Serum PBUT concentrations increased with advancing CKD stage. The mean MMSE score was 26.7 ± 3.4, and scores progressively declined with advancing CKD stage. A higher PBUT score was independently associated with MCI (coefficient, 0.290 ± 0.077; odds ratio, 1.337; 95% confidence interval, 1.151–1.554; p < 0.001). In conclusion, a greater PBUT burden was independently associated with cognitive impairment. These findings identify PBUT burden as a potential correlate of cognitive impairment in CKD. Prospective studies are needed to clarify its temporal relationship with cognitive decline and to assess whether it provides predictive value beyond established measures of kidney function.

Toxins doi: 10.3390/toxins18100421

Authors: Ibraimo Teleha Chabite Jamal Bakari Kussaga Frida Nyamete Quenton Kritzinger Shivan Maharaj Rashid Suleiman

Aflatoxin contamination in maize remains a major food safety and public health concern in Nampula Province, Mozambique. Maize samples obtained from farmers, traders, and milling units across the districts of Malema, Mossuril, Mecuburi, Monapo, and Nampula in 2024 and 2025 were analysed using UPLC-ESI-MS/MS. In 2024, aflatoxins were detected in samples from Mecuburi (43%), Mossuril (17%), Monapo (17%), and traders (13%), with some exceeding the regulatory limit of 15 µg/kg. AFB1 was the predominant aflatoxin, followed by AFB2. At the farmer level, Mecuburi recorded the highest contamination, with AFB1 and AFB2 detected in 43% of samples and AFG1 and AFG2 in 29%, while Mossuril had 20% contamination with AFB1 and AFG1. The maximum concentration was 4221.33 µg/kg of AFB1 in Mecuburi, whereas the minimum detected concentration was 2.82 µg/kg of AFB2. Aflatoxins were not detected above the limit of detection in milling samples or in samples collected in 2025. Despite the relatively low prevalence, the occurrence of extremely high aflatoxin concentrations highlights persistent contamination risks and the need for integrated mitigation strategies throughout the maize supply chain.

Toxins doi: 10.3390/toxins18100420

Authors: Guoping Li Lingli Meng Siyuan Feng Pengjuan Cui Gemei Liang Kongming Wu Qiuju Qin Hongqiang Feng

Mythimna (=Leucania) loreyi (Lepidoptera: Noctuidae) is an important pest of summer maize (Zea mays) in China’s Huang–Huai–Hai region. The approved transformation events DBN9936 (Cry1Ab) and Ruifeng 125 (Cry1Ab/Cry2Aj) were primarily developed to target Ostrinia furnacalis, and their efficacy against other lepidoptera pests, such as M. loreyi, remains largely unknown. Using age-stage two-sex life tables under controlled conditions, we quantified lethal and sublethal effects on these transformation events on first- to third-instar M. loreyi larvae fed V6-V8 leaves, and further evaluated how host plant movement scenarios affected larval development. Within a 7-day exposure period, both transformation events exerted significant lethal effects on M. loreyi. Corrected mortalities for first- to third-instar larvae were 93.43%, 17.33%, and 8.61% for DBN9936, and 44.33%, 8.53%, and 3.34% for Ruifeng 125, respectively. Individuals that survived on Bt maize exhibited significant sublethal effects, including prolonged larval development (ΔT = 8.12~9.93 d), reduced pupation rates (decreased by 69.41~100.00%), lower pupal mass (reduced by 31.14~33.64%), and decreased fecundity (by 177.86~262.96 eggs/female). Life table analysis revealed that both transformation events suppressed M. loreyi population growth in an instar-dependent manner. First-instar larvae failed to survive on either Bt transformation event. For second-instar larvae, DBN9936 remained completely lethal, while Ruifeng 125 allowed minimal survival (R0 = 7.51, relative fitness = 0.03). For third-instar larvae, partial survival occurred on both Bt events (DBN9936: R0 = 21.68, relative fitness = 0.09; Ruifeng 125: R0 = 32.34, relative fitness = 0.14), with rm values (0.06–0.07), lower than those on conventional controls (0.14–0.16). Host plant movement scenarios also affected M. loreyi survival and mass gain in an age-dependent manner. Brief early exposure to Bt maize did not reduce survival, but significantly suppressed body mass, whereas continuous Bt exposure caused drastic mortality in young larvae (2.08–8.33% survival). Older larvae tolerated Bt maize without survival loss, yet still exhibited significantly reduced body mass (p < 0.05). These findings demonstrate that DBN9936 and Ruifeng 125 exert substantial significant lethal and sublethal effects on M. loreyi, but the ability of older larvae to survive on Bt maize, combined with potential larval movement between Bt and non-Bt plants may facilitate resistance evolution.

Toxins doi: 10.3390/toxins18100419

Authors: Martina Ferrillo Claudio Curci Dario Calafiore Lorenzo Lippi Yasemin Yumuşakhuylu Michele Vecchio Vincenzo Quinzi Alfredo De Rosa Marco Invernizzi Alessandro de Sire

Temporomandibular disorders (TMDs) are heterogeneous musculoskeletal conditions frequently characterized by pain, impaired mandibular function, and reduced quality of life. Botulinum neurotoxin type A (BoNT-A) has emerged as a potential therapeutic option, particularly for persistent myogenous TMDs associated with masticatory muscle hyperactivity. This narrative review critically examines the mechanisms of action, current clinical evidence, therapeutic applications, and safety profile of BoNT-A in TMD management. Available evidence suggests that BoNT-A can reliably reduce masticatory muscle activity and may provide pain relief in selected patients with refractory myogenous TMDs; however, its superiority over placebo or established conservative treatments remains inconsistent. Considerable heterogeneity in patient selection, injected muscles, dosing regimens, injection techniques, and outcome assessment currently prevents the definition of standardized treatment protocols. Although short-term adverse events are generally mild and transient, repeated injections may induce muscle atrophy, reduced bite force, and potentially mandibular skeletal changes, whose long-term clinical relevance remains uncertain. Therefore, BoNT-A should not be considered a first-line treatment but rather a carefully selected adjunct within a multimodal rehabilitation strategy after appropriate conservative management has failed. Future phenotype-stratified comparative trials with standardized protocols and long-term structural safety outcomes are warranted.

Toxins doi: 10.3390/toxins18100418

Authors: Roko Gladović João G. Brás Jeroen C. Vollenbroek Karin G. F. Gerritsen

Human serum albumin (HSA) is the most abundant plasma protein and plays essential roles in regulating oncotic pressure, transporting endogenous and exogenous compounds, and maintaining redox balance. Because of its biochemical properties, HSA is widely used in clinical medicine, biotechnology, and pharmaceutical development. However, albumin is not a static molecule. It undergoes numerous post-translational modifications (PTMs), including oxidation, glycation, carbonylation, carbamylation, and aggregation, which can significantly alter its structure, ligand-binding properties, antioxidant capacity, and pharmacokinetics. Furthermore, commercial albumin products exhibit substantial heterogeneity arising from differences in source material, manufacturing processes, formulation conditions, and storage practices. This review summarizes the current understanding of PTMs affecting human serum albumin, with particular emphasis on their molecular origins and functional implications. We discuss how physiological, pathological, and manufacturing-induced modifications influence albumin activity and contribute to variability among commercial albumin products. In addition, we examine established and emerging applications of PTM-modified albumin in diagnostics, critical care, extracorporeal detoxification, drug delivery, nanomedicine, and regenerative medicine. Finally, we highlight current challenges related to product heterogeneity, quality control, regulatory assessment, and discuss future opportunities in albumin-based technologies. A deeper understanding of albumin PTMs is essential for improving critical care and developing next-generation albumin-based biomedical products.

Toxins doi: 10.3390/toxins18100417

Authors: Gianpaolo Ronconi Rossella Calciano Alberto Cutaia Rosa Cioffi Eslam Abdelkhalek Farag Anna Sanfilippo Martina Ferrillo Alessandro de Sire Paola Emilia Ferrara

Temporomandibular disorders (TMDs) represent a heterogeneous group of conditions, including myogenous and arthrogenous, with a significant impact on quality of life. Botulinum toxin type A (BoNT-A) has been proposed as a therapeutic option, particularly for patients with refractory myogenous TMD and bruxism. This systematic review aimed to evaluate the clinical effectiveness of BoNT-A in the management of TMDs overall and bruxism, with particular focus on myogenous TMDs compared to placebo and non-surgical treatments. RCTs published between 2020 and 2025 were identified through PubMed, Scopus, and Google Scholar. Overall, 15 RCTs involving 797 participants were included. The available evidence suggests potential benefits of BoNT-A, particularly in patients with localized myofascial pain, myogenous TMD, or marked masticatory muscle hyperactivity. Interpretation of the findings is limited by substantial heterogeneity in treatment protocols, toxin dosage, diagnostic criteria, outcome measures, and follow-up duration. Current evidence does not support the routine use of BoNT-A as a first-line treatment for TMDs overall. Instead, BoNT-A may represent an adjunctive or second-line option for selected patients with myogenous TMD, localized myofascial pain, or marked masticatory muscle hyperactivity who do not adequately respond to conventional noninvasive treatment. Larger and methodologically rigorous RCTs are needed to establish appropriate indications, duration of treatment effects, and long-term safety across different TMD subtypes.

Toxins doi: 10.3390/toxins18100416

Authors: Marta Mira Jørs Geoffrey Masuyer René Jørgensen

Diphthamide-specific mono-ADP-ribosylating toxins form a structurally conserved but phylogenetically diverse family of AB-type bacterial toxins, with Pseudomonas exotoxin A (PE) serving as a prototypical member. Here, we expand the known diversity of PE-like toxins (PLTs) through comprehensive genome mining and phylogenetic, structural and sequence analyses of PE, Chx, and their Aeromonas and Vibrio cholerae homologues. Aeromonas toxins separate into two distinct groups: a PE-like cluster (AEI) and a cholix-like cluster (AEII), while a fourth cholix subgroup (ChxIV) is identified within V. cholerae, expanding the previously described ChxI–III classification. Although all newly identified toxins retain the core catalytic architecture and hallmark residues essential for ADP-ribosylation of eEF2, they differ in key functional motifs, including receptor-binding residues, furin cleavage loops, and C-terminal ER-retrieval signals, suggesting variation in host-cell uptake routes and intracellular trafficking. Signal peptides were present in all toxins but varied modestly between clusters, consistent with normal species-level Sec-pathway variation rather than divergent secretion mechanisms. Together, our findings provide a significantly expanded overview of PLT diversity and indicate that Aeromonas-derived toxins comprise at least two evolutionarily and mechanistically distinct groups. These insights offer a framework for future functional studies and highlight PLTs as emerging virulence factors in environmentally and clinically relevant bacteria.

Toxins doi: 10.3390/toxins18100415

Authors: Kalina Zhivkova

Ochratoxin A (OTA), a mycotoxin produced by Aspergillus and Penicillium species, is a widespread feed contaminant with nephrotoxic, hepatotoxic, immunotoxic, reproductive, and teratogenic effects. Despite the particular susceptibility of rabbits to OTA, information on ochratoxicosis in this species remains limited. This review summarizes current evidence on OTA occurrence, toxicokinetics, mechanisms of toxicity, clinicopathological effects, immunotoxicity, and preventive strategies in rabbits. Following gastrointestinal absorption, OTA has a high affinity for plasma proteins and preferentially accumulates in the kidneys and liver. Its toxicity is mediated by the inhibition of protein synthesis, mitochondrial dysfunction, oxidative stress, genotoxicity, apoptosis, and immune dysregulation, ultimately leading to renal and hepatic injury, hematological and biochemical alterations, impaired growth, and reproductive toxicity. OTA-induced immunosuppression may also increase susceptibility to secondary infections, including Pasteurella multocida. Importantly, pathological and biochemical alterations may occur following relatively low-level exposure, while residual OTA levels may persist in tissues after exposure is discontinued. Effective prevention of OTA contamination in feed is therefore crucial for controlling ochratoxicosis. Microbial and enzymatic detoxification, probiotics, and phytogenic additives represent promising approaches for the prevention and mitigation of OTA toxicity; however, further studies in rabbits are needed to establish their efficacy and optimal application and to evaluate the effects of different OTA doses, early biomarkers, and tissue residue kinetics.

Toxins doi: 10.3390/toxins18100414

Authors: Qiaomin Duan Wanjun Zhang Ying Liu Sunlin Luo Wenjun He Wenjun Yang Yiqiang Chen

Aflatoxin B1 (AFB1) toxicity in poultry is often investigated using naturally contaminated ingredients, which can confound its effects with co-occurring mycotoxins and changes in feed quality. Male Arbor Acres broilers received a common basal diet supplemented with purified AFB1 at nominal concentrations of 0, 5, 10, 20, 50, 100, or 200 μg/kg for 42 d. The reported analyses detected no significant treatment differences in growth performance or major organ indices. Hepatic antioxidant enzyme activities and total antioxidant capacity decreased at selected exposures, while serum antioxidant enzymes, cytokines, and hematological indices showed selective responses. Jejunal villus height was lower in the nominal 50–200 μg/kg groups. Renal tubular lesions were most evident at 50–100 μg/kg. Bile-acid profiling in four selected groups showed compartment-specific differences. These findings suggest that selected biological and morphological responses may occur without detectable growth impairment under the conditions tested. Isolated biomarker differences remain exploratory, and the data do not establish a formal ranking of endpoint sensitivity or a dietary safety threshold.

Toxins doi: 10.3390/toxins18100413

Authors: Enrico Pambianco Octavian Augustin Mihalache Chiara Dall’Asta Antonia Garrido Frenich Laura Carbonell-Rozas

The aim of this study was to evaluate the co-occurrence and to assess the dietary health risk of multiple mycotoxins in fruit-based purées intended for infant consumption, combining targeted quantification and dietary exposure assessment with high-resolution suspect screening. Forty samples with varying apple content were randomly collected from Italian (n = 20) and Spanish markets (n = 20). A fit-for-purpose ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the simultaneous determination of patulin (PAT) and three Alternaria toxins, including alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN), providing satisfactory sensitivity, accuracy and precision. Targeted analysis revealed frequent co-occurrence of PAT, AOH, and AME, whereas TEN was not detected. Dietary exposure assessment indicated that PAT posed negligible health risks, whereas AOH and AME exceeded their respective Thresholds of Toxicological Concern (TTCs) across all evaluated population groups, indicating Alternaria toxins as the main contributors to dietary risk. High-resolution mass spectrometry (HRMS) suspect screening expanded the mycotoxin profile through the identification of tenuazonic acid, the tentative identification of altenusin and aflatoxin G2, and the annotation of 2,3-dihydroxybenzoic acid, a potential interferent in PAT determination. Overall, the proposed workflow provides a comprehensive strategy for mycotoxin monitoring in infant fruit purées and supports enhanced surveillance of emerging Alternaria toxins.

Toxins doi: 10.3390/toxins18100412

Authors: Marilena Caporale Giulia Marsico Patrizia Scafato Stefano Superchi

An efficient synthesis of phytotoxin (±)-3-deoxyradicinin was developed from commercially available 4-hydroxy-6-methyl-2H-pyran-2-one. The propenyl side chain was introduced through regioselective bromination followed by Horner–Wadsworth–Emmons olefination, while the second pyranone ring was constructed via a Lewis acid-catalyzed reaction with crotonic acid. The synthetic route was designed according to the principles of operational simplicity, with the aim of minimizing the use of hazardous reagents and solvents, as well as demanding purification procedures and cryogenic conditions. These objectives were successfully achieved for most of the synthetic sequence, with the use of CCl4 in the bromination step representing the main remaining limitation. Four of the six steps were successfully performed on a gram scale, demonstrating the practical potential of the route for a possible scale-up. In addition, a practical synthesis of the natural product 5,6-dehydrokawain was established. Overall, 319 mg of (±)-3-deoxyradicinin were obtained in 23% overall yield, providing a practical approach with reduced reliance on hazardous reagents and low-temperature conditions. This synthetic strategy therefore represents a useful platform for further scale-up of 3-deoxyradicinin preparation and for the synthesis of structurally related analogues.

Toxins doi: 10.3390/toxins18100411

Authors: Euikyung Kim

Animal venoms are among nature’s most sophisticated biochemical systems [...]

Toxins doi: 10.3390/toxins18100410

Authors: Marta Spochacz-Santoro Szymon Chowański Karolina Walkowiak-Nowicka Monika Szymczak-Cendlak Zbigniew Adamski

Glycoalkaloids are plant secondary metabolites known to affect multiple physiological processes in insects; however, their effects on carbohydrate and polyol metabolism remain poorly understood. This study investigated the effects of solasonine on hemolymph metabolites and aldose reductase activity in the fat body of Galleria mellonella larvae. Larvae were fed a diet supplemented with five nominal dietary concentrations of solasonine (10−4–1 mg/kg diet) for three days. Hemolymph metabolites were analyzed by RP-HPLC, and aldose reductase activity was determined in fat body lysates. Inositol levels differed significantly among experimental groups (one-way ANOVA, p < 0.0001), with the 10−4 mg/kg diet treatment differing significantly from the vehicle control (p = 0.0015). Aldose reductase activity was significantly lower at 10−2, 10−3, and 10−4 mg/kg diet compared with the untreated control (p = 0.0439, p = 0.027, and p < 0.0001, respectively), while only the 10−4 mg/kg treatment differed significantly from the vehicle control (p = 0.0029). Formic acid also significantly affected several metabolic parameters. Overall, solasonine exposure was associated with altered aldose reductase activity and changes in polyol-related metabolites, although some metabolic effects may be attributable to the vehicle.

Toxins doi: 10.3390/toxins18100409

Authors: Bryan G. Fry

Debate over the origin of squamate venom is frequently distorted by treating molecular availability, toxin recruitment and secretion, functional venom, and delivery-system elaboration as a single evolutionary event. This review separates these transitions and reassesses early venom evolution in anguimorph lizards and snakes using anatomical, molecular, proteomic, functional, ecological, and phylogenetic evidence. The species-level ancestral-state reconstructions of a recent Wüster, Modahl, and Arbuckle manuscript are internally robust to the four coding schemes tested and accommodate uncertain tips and reversible venom loss. Their inference nevertheless remains conditional on a binary definition of observed functional venom, a single hidden precursor category, a homogeneous transition process, and non-caenophidian snake states that were not varied in the relevant sensitivity analyses. While they coded two origins of venom within Anguimorpha, gland anatomy and derived oral products make an early functional origin followed by retention, reduction, loss, and parallel elaboration a strongly supported alternate hypothesis. However, as critical bridge taxa lack direct evidence of natural delivery, realistic dose, or secretion-specific fitness benefit, resolution of the history remains unresolved. For Heloderma, field studies of H. suspectum and H. horridum, together with painful, prolonged defensive bites, support a defensive role, although its primacy (to the exclusion of an additional predatory function) as advocated by them is sensitive to observational bias. Neurotoxic, cardiovascular, and coagulotoxic activities make a predatory contribution supported, leaving the original selective driver unresolved. Within Serpentes, their placement of venom at the base of the Caenophidia is challenged by distributed maxillary, mandibular, and rictal glands and derived oral products in non-caenophidian taxa, making an origin before Caenophidia a viable alternative hypothesis. Derived mucous systems in boas and pythons and specialised Scolecophidia cannot alone polarise the ancestral snake state. Current evidence is therefore insufficient to distinguish one shared functional origin from two functionalisations of a homologous ancestral oral secretory system. No alternative comparative reconstruction is performed; the review shows only that earlier origins remain viable and categorical absence coding is insufficiently secure for several poorly investigated taxa. Resolution requires matched gland cell biology, orthology- and synteny-resolved genomics, quantitative secretion proteomics, realistic functional assays, secretion-specific ecological tests, broader Iguania and non-caenophidian snake sampling, and ancestral-state models that represent graded evidence, both false-negative and false-positive classification, reduction, loss, and phylogenetic uncertainty.

Toxins doi: 10.3390/toxins18100408

Authors: Maxine Bennek Martina Kerscher

Botulinum neurotoxin type A (BoNTA) is widely used for the treatment of dynamic facial wrinkles, where precise control of toxin diffusion is essential to achieve predictable aesthetic outcomes while preserving natural facial expression. However, the relative influence of injection pattern, dose, and dilution on diffusion characteristics remains incompletely understood. In this exploratory case series, treatment-naïve subjects undergoing aesthetic treatment of horizontal forehead lines with LetibotulinumtoxinA (Leti-BoNTA) were assigned to one of four injection paradigms according to their wrinkle pattern: a conventional five-point injection pattern, the previously published LEBO injection pattern, dose variation (2–5 U per injection point), or dilution variation (reconstitution volumes 0.675–2.5 mL). Diffusion was assessed using the Minor iodine–starch test, and anhidrotic areas were quantified in cm2. Increasing dose was associated with progressively larger anhidrotic areas in this exploratory case series. Greater reconstitution volumes were associated with broader toxin distribution, whereas lower dilution volumes produced more confined diffusion areas. The conventional five-point injection pattern resulted in relatively extensive diffusion, whereas the LEBO protocol demonstrated a more localized and consistent distribution profile. Despite differences in diffusion characteristics, natural facial dynamics were preserved across all treatment paradigms. These findings suggest that injection pattern, dose, and dilution collectively influence Leti-BoNTA diffusion and should be considered when optimizing individualized treatment strategies.

Toxins doi: 10.3390/toxins18100407

Authors: Yuan Guo Yuxin An Jiaojiao Xu Zengxuan Cai Meijia Xu Xiaomin Xu

Bongkrekic acid (BA) is a metabolite that is lethal to humans and is produced by Burkholderia gladioli pathovar cocovenenans (BGC) containing the bon gene cluster. An analytical method for BA and its stereoisomer, iso-bongkrekic acid (IBA), was established for deeply fermented rice products, such as red yeast rice and its related products, by liquid chromatography–tandem mass spectrometry (LC-MS/MS). BA and IBA in samples were extracted with 80% methanol/water containing 1% ammonia by ultrasonic assistance, cleaned by solid-phase extraction (SPE) with an anion-exchange cartridge, separated with an XBridgeTM BEH C18 chromatographic column (150 mm × 3.0 mm, 2.5 μm), measured by MS/MS, and quantified using an isotope-labeled internal standard. The influencing factors such as extraction solvents, ultrasonic extraction time and loading volume during SPE clean-up were optimized to improve the recoveries of BA and IBA. Also, the extraction efficiency between the simulated quality control and directly spiked samples, the matrix effects with the use of different chromatographic columns, the impact of co-eluting interferences under different mobile phase compositions and the conversion of BA to IBA are discussed. The linear range of BA and IBA was 0.1–50 ng/mL (equivalent to 1–500 μg/kg in the sample), with the correlation coefficient exceeding 0.999. The limits of detection and quantification were 0.3–0.5 μg/kg and 1.0–1.5 μg/kg, respectively. The average recoveries of the analytes were 88.4–107% with relative standard deviations (RSDs, n = 6) between 2.9% and 11.8% in red yeast rice and its related products under three spiking levels. This optimized method was applied to study the contamination of BA and IBA in commercially available samples. This study can provide technical support for the supervision and risk assessment of BA and IBA in red yeast rice and its related products.

Toxins doi: 10.3390/toxins18100406

Authors: Rozenn Souillard Sandra Rouxel Virginie Dorenlor Grégoire Kuntz Félix Mahé Typhaine Poezevara Florent Eono Amandine Avouac Stéphane Kerphérique Léa Jambou Eloïse Cailton Mathys Gouet Sébastien Solanas Loïc Balaine Adeline Huneau-Salaün Marianne Chemaly Sophie Le Bouquin Alain Hartmann Caroline Le Maréchal

Animal botulism mainly affects poultry and cattle, with risks of cross-contamination. This study aimed to evaluate Clostridium botulinum contamination in manure and soil following manure spreading on 13 clinically affected farms (10 cattle and three poultry) and 39 clinically unaffected farms (20 cattle and five poultry farms without any previous botulism outbreak, six poultry farms with a history of botulism and eight mixed poultry/cattle farms with an ongoing bovine botulism outbreak). Twelve manure samples, two swabs on a manure pile, 12 soil samples, and two boot swabs from fields were collected before manure spreading. Twelve soil samples and two boot swabs from fields were obtained after manure spreading. C. botulinum types C, D, C/D, and D/C were detected using real-time PCR after an enrichment step. On clinically unaffected poultry and cattle farms with no history of botulism, C. botulinum was detected in 1.4% and 1.8% of samples, respectively. Detection rates reached 8% on poultry farms with a history of botulism and 42.8% on poultry farms with ongoing bovine outbreaks. On clinically affected poultry and cattle farms, 34% and 18.9% of samples were positive, respectively. This study highlights the presence of C. botulinum in manure and soil and emphasizes the importance of biosecurity measures to prevent cross-contamination and disease recurrence.

Toxins doi: 10.3390/toxins18090405

Authors: Tulio Custódio Reis Ana Clara Lunardi Yagi Eloise T. Filardi Marcela Romanazzi Michele Ferreira da Silva Mela Rui S. Ferreira Júnior Felipe A. Cerni Márcia A. S. Graminha Manuela B. Pucca

Leishmaniasis and Chagas disease are neglected tropical diseases that continue to pose significant public health challenges, highlighting the need for new therapeutic strategies. Brazilian snake venoms represent a valuable and underexplored source of bioactive molecules with diverse pharmacological activities and promising therapeutic potential. In this context, the present study aimed to investigate the in vitro antiprotozoal potential of venoms from Bothrops and Crotalus species, including Bothrops jararacussu, Bothrops pauloensis, Bothrops alternatus, Bothrops leucurus, Bothrops atrox, Bothrops jararaca, Crotalus durissus terrificus, and Crotalus durissus ruruima, metalloprotease (MTL)- and crotoxin (CTX)-enriched, against Leishmania spp. and Trypanosoma cruzi. Crude venom protein profiles were analyzed by SDS-PAGE, and cytotoxicity was assessed in differentiated C2C12 myotubes and murine peritoneal macrophages. Antiprotozoal activity was evaluated against extracellular parasite forms, including promastigotes of Leishmania amazonensis and Leishmania infantum and epimastigotes of T. cruzi. Venoms exhibiting the highest selectivity indices (SIs), calculated using murine macrophage cytotoxicity data, were further tested against intracellular amastigotes of L. amazonensis. Among the tested samples, B. jararacussu venom showed activity against promastigotes of both L. amazonensis (IC50 = 2.9 µg mL−1) and L. infantum (IC50 = 2.8 µg mL−1), as well as in epimastigotes of T. cruzi (IC50 = 15.5 µg mL−1). The MTLand CTX pools from C. d. ruruima venom exhibited the highest activity against promastigotes of L. amazonensis, with IC50 values of 1.6 and 3.8 µg mL−1, respectively. Against intracellular amastigotes, MTL and CTX showed IC50 values of 4.1 and 2.5 µg mL−1, respectively, with selectivity indices (SIs) of 73.1 and 120, respectively. Overall, these findings provide preliminary evidence for the antiparasitic potential of snake venom-derived molecules, supporting further characterization and in vivo evaluation.

Toxins doi: 10.3390/toxins18090404

Authors: Kazuya Yoshida

Botulinum neurotoxin (BoNT) therapy has developed from a treatment primarily used for neurological disorders into a versatile therapeutic modality applied across a broad range of medical and dental conditions [...]

Toxins doi: 10.3390/toxins18090403

Authors: Andrea Felice Armenti Francesco Armenti

Pharmacodynamic models of botulinum toxin type A map a mean concentration onto a smooth dose–response curve, yet the outcome is binary and arises from a finite population of near-threshold motor units. We analyse a discrete model in which N units are silenced once local SNAP-25 cleavage exceeds a unit-specific threshold, and a functional block of neuromuscular transmission follows once the silenced fraction exceeds a collective threshold. Whether that collective threshold is a fraction of the population or a fixed count changes the location of the dose–response curve and not its steepness: under a fraction, potency is invariant to unit number at fixed dose–concentration gain, and the transition contracts in absolute dose where fixed-count architectures require it to widen. The width contracts as N−1/2, an exponent that is prior and not claimed here. One consequence needs no measurement: a quantal terminal response caps the silenced fraction below unity, so any fixed count fails above a target size it determines, whereas the toxin block targets differ by orders of magnitude. A second reinterprets existing data, flat cohort dose–response curves being what a near-step individual response predicts once convolved with between-subject dispersion. Outcome is accordingly less repeatable in the smallest targets. No parameter is fitted.

Toxins doi: 10.3390/toxins18090402

Authors: Bart Eeckhaut Steven Truijen Parham Haghshenas Antriana Roussou Petros Taflampas Wim Saeys

Background: Botulinum toxin type A (BoNT-A) is globally recognized as a standard treatment for post-stroke spasticity (PSS), although its secondary mechanisms remain underinvestigated. The objective of this study was to investigate the neurophysiological mechanisms underlying BoNT-A treatment in PSS and explore their implications for mechanism-based rehabilitation. Methods: A systematic review and meta-analysis (PROSPERO registration number ID: CRD420261352230) of 39 studies involving 760 patients was conducted on 18 August 2026, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. PubMed, Web of Science, Scopus, and Embase were systematically searched using the PICO framework. Where possible, quantitative data were pooled according to predefined neurophysiological constructs, including passive stretch-evoked activity, voluntary activation of the injected muscle, compound muscle action potential (CMAP), reciprocal motor control, normalized Hmax/Mmax, and specific spinal inhibitory mechanisms. Results: Quantitative synthesis demonstrated construct-specific neurophysiological changes following BoNT-A treatment. Passive stretch-evoked activity showed the largest pooled effect (Hedges’ g = 0.80, 95% CI 0.35–1.24), followed by CMAP amplitude (g = 0.69, 95% CI 0.23–1.14) and reciprocal motor control (g = 0.52, 95% CI 0.23–0.81). Effects on voluntary activation of the injected muscle (g = 0.31, 95% CI −0.38 to 1.00) and normalized Hmax/Mmax (g = 0.22, 95% CI −0.74 to 1.17) were smaller and more heterogeneous. Reciprocal and recurrent inhibition were each represented by single-study estimates, while supraspinal outcomes were synthesized narratively due to methodological heterogeneity. Conclusions: The findings support a predominantly peripheral mechanism of BoNT-A while suggesting possible secondary changes within spinal and supraspinal motor control pathways. However, the certainty of evidence was low to very low. These neurophysiological effects provide a rationale for individualized, mechanism-based rehabilitation during the post-injection therapeutic window, although higher-quality evidence is required to confirm this framework.

Toxins doi: 10.3390/toxins18090401

Authors: Corinne M. Gibble Melissa A. Miller Raphael M. Kudela Angelina Reed Kevin Keel Rebecca S. Duerr

In 2017, an unusually large number of live stranded and dead marine birds were found in California, coincident with a large bloom of the diatom Pseudo-nitzschia that produced the toxin domoic acid (DA). The most abundant bird species affected by this large die-off and stranding event were Pacific Loons (Gavia pacifica) and Red-throated Loons (Gavia stellata). International Bird Rescue (IBR; San Pedro, CA) received 117 live loons with suspected DA toxicosis. Many birds that survived the first few days in rehabilitation quickly re-attained suitable health metrics; however, 68 loons (58.1%) died or were considered unreleasable due to ongoing abnormal behavior or injuries that necessitated humane euthanasia. In this descriptive study, both live stranded euthanized birds and dead stranded birds were examined to characterize and better understand the effects of DA toxicosis in the avian brain and body. Utilizing a suite of diagnostics, our analysis revealed novel pathological patterns that divided these birds into three groups: those suspected of having acute, subacute, and chronic DA toxicosis. Domoic Acid was found in all groups, and the severity of the microscopic lesions appeared to correspond with the amount of time each bird survived post-stranding. Our investigation is the first known account identifying the pathological progression from acute to chronic DA toxicosis in birds.

Toxins doi: 10.3390/toxins18090400

Authors: Riccardo Marvulli Serena Caforio Montesardo Chiara Moccia Marisa Megna Maurizio Ranieri

Upper limb spasticity is a common complication of stroke that negatively affects motor function, range of motion, and quality of life. Although abobotulinumtoxinA is widely used for the treatment of focal post-stroke spasticity, evidence regarding its early onset of action remains limited. This prospective observational pilot study investigated the early clinical, functional, and neurophysiological effects of abobotulinumtoxinA in 20 adults with post-stroke upper limb spasticity. Patients received ultrasound-guided injections into the biceps brachii and flexor digitorum superficialis muscles and were evaluated at baseline, 24 h, 5 days, 7 days, 14 days, and 30 days after treatment. Outcome measures included the Modified Ashworth Scale (MAS), passive range of motion (ROM), caregiver-reported hand function assessed using a visual analog scale (VAS), and compound muscle action potential (cMAP) recordings. No significant changes were observed at 24 h. A significant reduction in spasticity was detected at the day-5 assessment, accompanied by significant improvements in ROM and functional outcomes. Neurophysiological assessment demonstrated significant reductions in cMAP amplitudes at the day-5 assessment, indicating early neuromuscular transmission blockade. Clinical improvements were also first detected at day-5 assessment and were maintained at subsequent evaluations through day 30. These findings indicate that clinically meaningful effects of abobotulinumtoxinA were detectable by the first scheduled assessment at day 5 after injection. Given the absence of intermediate assessments between 24 h and day 5, the exact timing of treatment onset cannot be determined. A multimodal assessment approach may help characterize early treatment-related changes and provide useful information for future studies investigating the timing of adjunctive rehabilitation interventions.

Toxins doi: 10.3390/toxins18090399

Authors: Audra Janovskienė Justina Stučinskaitė-Maračinskienė Žygimantas Petronis Jan Pavel Rokicki Dainius Razukevičius Loreta Pilipaitytė

Background: Botulinum toxin type A (BoNT-A) has been proposed as an adjunct to orthognathic surgery because its temporary neuromuscular effects may reduce muscular loading during postoperative healing and potentially influence fixation stability, skeletal relapse, and postoperative recovery. This systematic review aimed to evaluate the available clinical evidence regarding the adjunctive use of BoNT-A in orthognathic surgery. Methods: A systematic literature search was conducted in PubMed, the Cochrane Library, and Google Scholar in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Clinical studies directly comparing orthognathic surgery with and without adjunctive BoNT-A administration and reporting outcomes related to mechanical stability, skeletal stability, or postoperative recovery were included. Risk of bias was assessed using the Cochrane Risk of Bias 2 (RoB 2) tool for randomized controlled trials and ROBINS-I for the non-randomized study. Results: Three studies met the eligibility criteria, comprising two randomized controlled trials and one retrospective comparative study. Considerable heterogeneity was observed in surgical procedures, targeted muscles, BoNT-A doses, timing of administration, and evaluated outcomes. Individual studies reported associations between BoNT-A administration and a lower incidence of fixation plate fracture, reduced anteroposterior skeletal relapse at the Pogonion, or lower preanalgesic pain scores and postoperative opioid consumption. However, these findings were derived from different studies evaluating distinct interventions and outcomes and were not consistently demonstrated across comparable outcome measures. Conclusions: BoNT-A may represent a promising adjunct to orthognathic surgery, with potential benefits for fixation-related mechanical stability, selected parameters of skeletal relapse, and postoperative pain management. However, the limited number of studies, methodological limitations, and substantial clinical heterogeneity preclude definitive conclusions. Further well-designed randomized controlled trials with standardized injection protocols and longer follow-up are required before routine clinical use can be recommended.

Toxins doi: 10.3390/toxins18090398

Authors: Aideen Steed Heather Williams Jessie Alfonso Denise Hancock Stephen A. Ashford

Botulinum toxin type A (BoNT-A) is an established treatment for spasticity. Goal Attainment Scaling (GAS), alongside standardized measures such as Arm Activity Measure (ArmA) and Leg Activity Measure (LegA), enables evaluation of patient-centered outcomes. This real-world cohort study evaluated goal attainment and standardized measure functional changes following BoNT-A for arm and/or leg spasticity, evaluated using GAS as well as passive function subscales of ArmA (ArmA-PF) and LegA (LegA-PF), and active function subscales (ArmA-AF; LegA-AF). Inclusion in the analysis was based on documented baseline and post-intervention GAS scores following BoNT-A therapy (n = 390). Across 855 treatment cycles, 1258 goals were set and categorized, of which 150 (11.9%) were related to active function and 705 (56.0%) to passive function. The mean GAS T-score improved from 36.2 to 49.4 (change 13.1, p < 0.001), with significant gains observed for both passive (change 13.7, p < 0.001) and active goals (change 12.6, p < 0.001). Passive function improved significantly on ArmA-PF (change 2.5) and LegA-PF (change 2.1), while significant changes in ArmA-AF and LegA-AF were not seen. In conclusion, goal-directed spasticity management incorporating BoNT-A resulted in improvements in goal attainment, which corresponded to a clinically meaningful change in passive function measured by ArmA-PF and LegA-PF, but not for active function. Passive function was the most common treatment priority. Findings provide real-world evidence to support community-based spasticity services and the use of structured goal setting, ArmA and LegA application, in routine practice.

Toxins doi: 10.3390/toxins18090397

Authors: Noah M. Souza Alexander O. Hale Richard S. Beard Juliette K. Tinker

AB5 toxins are important virulence factors produced by Gram-negative bacteria and have been identified in many non-typhoidal Salmonella (NTS) serovars; however, their effects on host cell signaling and epithelial barrier function remain poorly understood. This study characterized the molecular and functional effects of purified ArtAB toxin and its binding subunit, ArtB, from multidrug-resistant Salmonella Typhimurium definitive type 104 (DT104). Equimolar concentrations of ArtAB (10 µg/mL) and ArtB (7.25 µg/mL) were evaluated in Chinese hamster ovary (CHO-K1) cells using cytotoxicity assays, quantitative proteomics, and mRNA sequencing, while barrier function was assessed in C2BBe1 human intestinal epithelial cells using electric cell-substrate impedance sensing (ECIS). In CHO-K1 cells, both proteins exhibited minimal cell death. ArtB elicited pronounced proteomic changes associated with endoplasmic reticulum (ER) homeostasis, vesicular trafficking, and lysosomal processing, but limited transcriptional responses. ArtAB instead altered host transcriptional programs associated with cell growth, inflammatory signaling, and cytoskeletal organization. Unlike ArtB, ArtAB significantly reduced barrier function in C2BBe1 cells. Together, these findings show that ArtAB remodels host cell signaling without overt cytotoxicity in CHO-K1 cells and produces a distinct functional effect in C2BBe1 cells. Although obtained in different cellular models, these results provide insight into the potential role of ArtAB in NTS pathogenesis.

Toxins doi: 10.3390/toxins18090396

Authors: Guangyao Ying Maofa Zeng Liang Hong Zhaokui Li Jun Li Huili Xia

Mycotoxin contamination represents one of the most pressing food safety challenges worldwide, with millions of tons of agricultural commodities affected annually. Conventional detection methods, while accurate, are labor-intensive, destructive, and unsuitable for large-scale screening. Hyperspectral imaging (HSI) has emerged as a transformative non-destructive analytical technique capable of simultaneously capturing spatial and spectral information across hundreds of contiguous wavelengths. This review critically evaluates the current state of HSI technology for mycotoxin detection in food products, covering the physical principles underlying spectral–mycotoxin interactions, systematic applications across major mycotoxin classes (aflatoxins, deoxynivalenol, ochratoxin A, fumonisins, and zearalenone), and the integration of machine learning and deep learning algorithms for spectral data analysis. The review reveals that while Vis-NIR (400–1000 nm) and SWIR (1000–2500 nm) HSI systems have achieved classification accuracies exceeding 90% for several mycotoxin–matrix combinations, fundamental challenges persist in model transferability, direct quantification at regulatory thresholds, and scalability for industrial deployment. Recent advances in transformer architectures, transfer learning, interpretable deep learning, and portable multispectral systems demonstrate encouraging progress toward practical implementation. This review concludes by identifying critical research gaps and proposing strategic directions for translating HSI-based mycotoxin detection from laboratory proof-of-concept to routine industrial application.

Toxins doi: 10.3390/toxins18090395

Authors: Galina Nugumanova

Cyanobacterial harmful algal blooms (cyanoHABs) typically involve the release of a wide spectrum of toxic metabolites produced by different cyanobacteria, which can cause adverse health effects in animals and humans. At the same time, many cyanobacteria are capable of producing more than one toxin simultaneously. As a result, the multiclass toxins released into aquatic environments can exert complex and combined effects on various ecosystem components. This review aims to provide an overview of the occurrence and effects of different cyanotoxins in aquatic systems and current analytical methods of their simultaneous detection. The main classes of cyanotoxins have been considered in this review. The occurrences, properties, and environmental and health impacts of these toxins are described. The main methods for cyanobacterial toxin analysis are covered. A comprehensive review and analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection and sample preparation approaches used to identify various classes of cyanotoxins is provided, together with a practical decision tree for analytical workflow selection according to toxin class and sample matrix, highlighting key trends and outlining directions for further research in this area.

Toxins doi: 10.3390/toxins18090394

Authors: Yong-Hua Wu Zheng-Xu Zhong Yan Li Jing-An Wang Zheng-Wen Ou Hou-Jin Li Wen-Jian Lan

Vespa basalis Smith venom (VBsV) causes painful and potentially severe systemic reactions, yet the relationships among its composition, bioactivity, and cytotoxicity remain poorly understood. Proteomic and metabolomic profiling was combined with ultrafiltration and gel filtration to characterize VBsV and obtain >3 kDa, operationally defined 700–3000 Da, and <700 Da fractions. The effect on LPS-induced NO production was evaluated by nitric oxide (NO) release in LPS-stimulated RAW264.7 cells, and cytotoxicity was assessed in tumor and THLE-2 liver cells. Five venom allergen proteins were identified, including two phospholipase A1 isoforms, hyaluronidase A, antigen 5, and venom dipeptidyl peptidase IV. Targeted profiling also detected several neurotransmitters and energy metabolites, including 5-hydroxytryptamine, L-glutamate, histamine, γ-aminobutyric acid, inosine, and adenosine. The 700–3000 Da fraction exhibited the strongest NO-suppressive and tumor-cell-viability effects among the tested fractions. The 700–3000 Da fraction suppressed NO release (IC50, approximately 23 μg/mL) and reduced tumor-cell viability, while minimally affecting THLE-2 cells at 10–80 μg/mL; by contrast, the >3 kDa fraction showed greater THLE-2 cytotoxicity. These findings identify the 700–3000 Da fraction as a priority for direct compositional characterization and isolation of the responsible molecules.

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.