Toxins

21 pages, 6401 KiB

Open AccessArticle

The Dissociation of Latrophilin Fragments by Perfluorooctanoic Acid (PFOA) Inhibits LTX^N4C-Induced Neurotransmitter Release

by Evelina Petitto, Jennifer K. Blackburn, M. Atiqur Rahman and Yuri A. Ushkaryov

Toxins 2025, 17(7), 359; https://doi.org/10.3390/toxins17070359 - 20 Jul 2025

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Abstract

α-Latrotoxin stimulates neurotransmitter release by binding to a presynaptic receptor and then forming ion-permeable membrane pores and/or stimulating the receptor, latrophilin-1, or Adhesion G-protein-coupled receptor type L1 (ADGRL1). To avoid pore formation, we use the mutant α-latrotoxin (LTX^N4C), which does not [...] Read more.

α-Latrotoxin stimulates neurotransmitter release by binding to a presynaptic receptor and then forming ion-permeable membrane pores and/or stimulating the receptor, latrophilin-1, or Adhesion G-protein-coupled receptor type L1 (ADGRL1). To avoid pore formation, we use the mutant α-latrotoxin (LTX^N4C), which does not form pores and only acts through ADGRL1. ADGRL1 is cleaved into an N-terminal fragment (NTF) and a C-terminal fragment (CTF), which behave as independent cell-surface proteins, reassociating upon binding LTX^N4C. We investigated the role of the NTF-CTF association in LTX^N4C action, using perfluorooctanoic acid (PFOA). We demonstrate that at low concentrations (≤100 μM) PFOA does not adversely affect ADGRL1-expressing neuroblastoma cells or inhibit LTX^N4C binding. However, it causes the dissociation of the NTF-CTF complexes, independent redistribution of the fragments on the cell surface, and their separate internalization. PFOA also promotes the dissociation of NTF-CTF complexes induced by LTX^N4C binding. When applied to mouse neuromuscular junctions, PFOA inhibits LTX^N4C-induced neurotransmitter release in a concentration-dependent manner. Our results indicate that ADGRL1 can mediate LTX^N4C signaling only while its fragments remain associated. These findings explain some aspects of receptor-dependent toxin action and contribute to a mechanistic understanding of ADGRL1 functions in neurons. Full article

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21 pages, 2039 KiB

Open AccessArticle

Comprehensive RNA-Seq Analysis of Human Osteoclast Function in Response to Bothrops moojeni Venom Fractions: Pathways of Bone Resorption and Cytoskeletal Disruption

by Fernanda D’Amélio, Hugo Vigerelli, Rodrigo Pinheiro Araldi, Isabel de Fátima Correia Batista, Daniel Carvalho Pimenta and Irina Kerkis

Toxins 2025, 17(7), 358; https://doi.org/10.3390/toxins17070358 - 19 Jul 2025

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Abstract

This study investigated the effects of Bothrops moojeni (B. moojeni) venom and its high- (HMM) and low-molecular mass (LMM) fractions on human osteoclast (OC) differentiation and function in vitro, aiming to identify novel therapeutics for bone disorders. Venom preparations were applied [...] Read more.

This study investigated the effects of Bothrops moojeni (B. moojeni) venom and its high- (HMM) and low-molecular mass (LMM) fractions on human osteoclast (OC) differentiation and function in vitro, aiming to identify novel therapeutics for bone disorders. Venom preparations were applied at 5 µg/mL (crude venom and HMM) or 1 µg/mL (LMM) from day 4 of peripheral blood mononuclear cell (PBMC) differentiation through terminal OC formation, enabling evaluation across early differentiation, fusion, and maturation stages. RNA sequencing revealed 7793 genes common to all experimental groups, with unique gene expression signatures of 149 (control), 221 (HMM), 248 (crude venom), and 60 (LMM) genes, reflecting distinct molecular responses. The negative control PBMC group exhibited 1013 unique genes enriched in immune-related pathways, consistent with their undifferentiated state. Crude venom induced the broadest transcriptional modulation, upregulating key fusion (CD47) and resorption (CTSK) genes, and altering markers of OC differentiation. The HMM fraction predominantly influenced inflammatory and osteoclastogenic pathways, notably TNF and NF-κB signaling, while the LMM fraction selectively regulated fusion-related genes (e.g., CD44) and immune pathways, indicating targeted modulation of OC activity. Cytokine profiling showed that crude venom and HMM suppressed osteoclastogenic cytokines such as IL-1β and IL-6, supporting their potential use in inflammatory bone diseases. Pathway enrichment analyses confirmed these differential effects on immune response and bone resorption mechanisms. Together, these results demonstrate that B. moojeni venom and its fractions differentially impact OC biology, with crude venom exerting broad effects and HMM and LMM fractions offering more specific modulation. Future studies will isolate bioactive components and assess therapeutic efficacy in animal models of osteoporosis and rheumatoid arthritis. Full article

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

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17 pages, 1217 KiB

Open AccessArticle

mRNA Expression of Two Colon Enzymes in Pre-Pubertal Gilts During a 42-Day Exposure to Zearalenone

by Magdalena Gajęcka, Łukasz Zielonka and Maciej T. Gajęcki

Toxins 2025, 17(7), 357; https://doi.org/10.3390/toxins17070357 - 17 Jul 2025

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Abstract

The aim of this study was to determine whether a low dose of zearalenone (ZEN) affects the mRNA expression of the CYP1A1 (P450 cytochrome) and GSTπ1 (glutathione S-transferase) genes in the large intestine of pre-pubertal gilts. Materials: Control (C) group gilts (n [...] Read more.

The aim of this study was to determine whether a low dose of zearalenone (ZEN) affects the mRNA expression of the CYP1A1 (P450 cytochrome) and GSTπ1 (glutathione S-transferase) genes in the large intestine of pre-pubertal gilts. Materials: Control (C) group gilts (n = 18) received a placebo. Experimental (E) group gilts (n = 18) were orally administered 40 μg ZEN/kg body weight (BW) each day before morning feeding for 42 days. Three animals from each group were sacrificed each week of the study. Tissue samples were collected from the medial parts of the ascending colon and the descending colon on six dates. Results: Zearalenone concentrations were multiple times higher in the last three weeks of exposure, and ZEN metabolites were not detected. In phase I, CYP1A1 mRNA expression in the ascending colon was suppressed in the final three weeks of exposure, which substantially increased the ZEN concentration in the descending colon. In phase II, ZEN levels were high in the descending colon due to CYP1A1 suppression in the ascending colon. Consequently, the phase II detoxification processes could not take place due to the absence of a substrate. Conclusion: This study demonstrated that low-dose ZEN mycotoxicosis disrupts the expression of the CYP1A1 and GSTπ1 genes, which co-participate in the enzymatic biotransformation of ZEN in both examined sections of the large intestine. The above could have contributed to increased ZEN accumulation in the mucosa of the descending colon in the last three weeks of exposure. Full article

(This article belongs to the Section Mycotoxins)

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21 pages, 385 KiB

Open AccessReview

Emerging Mycotoxins in Aquaculture: Current Insights on Toxicity, Biocontrol Strategies, and Occurrence in Aquafeed and Fish

by Patrizio Lorusso, Giusy Rusco, Alessio Manfredi, Nicolaia Iaffaldano, Angela Di Pinto and Elisabetta Bonerba

Toxins 2025, 17(7), 356; https://doi.org/10.3390/toxins17070356 - 17 Jul 2025

Viewed by 362

Abstract

Mycotoxins are secondary metabolites produced by various fungal species that can contaminate food and feed, posing significant risks to human and animal health. In aquaculture, the replacement of fishmeal with alternative protein sources has increased the risk of mycotoxin contamination, becoming a major [...] Read more.

Mycotoxins are secondary metabolites produced by various fungal species that can contaminate food and feed, posing significant risks to human and animal health. In aquaculture, the replacement of fishmeal with alternative protein sources has increased the risk of mycotoxin contamination, becoming a major challenge in fish feed production. Current data highlights that fish are exposed not only to common mycotoxins but also to emerging ones, raising concerns about human exposure through fish consumption. In this review, we draw attention to the toxicity data of key emerging mycotoxins from Fusarium (enniatins, ENNs; beauvericin, BEA) and Alternaria (alternariol monomethyl ether, AME; alternariol, AOH), their occurrence in aquafeeds and in commercially relevant fish species in Europe, and potential biocontrol approaches to prevent/mitigate contaminations. From the present review, it emerged that these mycotoxins exhibit in vitro cytotoxic properties. Their prevalence and concentrations vary widely both among aquafeeds, depending on the sample’s origin, and among fish species. Biocontrol approaches using microorganisms or natural compounds show promise as sustainable solutions to limit contamination. However, further research is essential to address data gaps and to allow for a proper risk assessment and, if necessary, the implementation of effective management measures. Full article

(This article belongs to the Special Issue Risk Assessment of Mycotoxins: Challenges and Emerging Threats)

10 pages, 598 KiB

Open AccessReview

Translational Impact of Genetics and Epigenetics of CGRP System on Chronic Migraine Treatment with Onabotulinumtoxin A and Other Biotech Drugs

by Damiana Scuteri and Paolo Martelletti

Toxins 2025, 17(7), 355; https://doi.org/10.3390/toxins17070355 - 17 Jul 2025

Viewed by 581

Abstract

Migraine is a neurovascular paroxysmal disorder characterized by neurogenic inflammation and has a remarkable impact on the quality of life. The Food and Drug Administration (FDA) approved onabotulinumtoxin A in 2010 for the prophylactic treatment of chronic migraine. Today, in its 4th decade, [...] Read more.

Migraine is a neurovascular paroxysmal disorder characterized by neurogenic inflammation and has a remarkable impact on the quality of life. The Food and Drug Administration (FDA) approved onabotulinumtoxin A in 2010 for the prophylactic treatment of chronic migraine. Today, in its 4th decade, it is approved in 100 countries for 15 main indications. Its mechanism of action, based on the inhibition of neurotransmitter release from primary sensory neurons, is very complex: it affords antinociception, but it also has an analgesic effect on neuropathic pain conditions and reduces the need for rescue medications. Genetic variants have been investigated for their potential role in the pathogenesis and clinical expression of migraine and of the response to treatments. These studies primarily involved genes associated with vascular regulation and cardiovascular pathology, including those encoding angiotensin-converting enzyme (ACE) and methylenetetrahydrofolate reductase (MTHFR). However, epigenetics and, particularly, genetic and epigenetic modifications are still poorly studied in terms of understanding the mechanisms implicated in susceptibility to migraine, aura, chronification and response to symptomatic and preventive treatments. In particular, the aim of the present study is to gather evidence on the genetic variants and epigenetic modifications affecting the pathway of the calcitonin gene-related peptide (CGRP), the target of onabotulinumtoxin A and of all the novel monoclonal antibodies. Full article

(This article belongs to the Special Issue Botulinum Toxins: Beyond the SNAP-25 Cleavage and New Insights on Pain Relief Mechanisms)

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24 pages, 3329 KiB

Open AccessArticle

Isolation of a Novel Streptomyces sp. TH05 with Potent Cyanocidal Effects on Microcystis aeruginosa

by Xuhan Wang, Siqi Zhu, Shenchen Tao, Shaoyong Zhang, Ruijun Wang and Liqin Zhang

Toxins 2025, 17(7), 354; https://doi.org/10.3390/toxins17070354 - 17 Jul 2025

Viewed by 475

Abstract

In this paper, cultivable actinobacteria were isolated, cultured, and identified from the heavily algal-bloomed waters of Taihu Lake using 16S rRNA gene sequencing. Among the isolates, a single strain exhibiting vigorous cyanocidal activity against Microcystis aeruginosa FACHB-905 was selected for further investigation. The [...] Read more.

In this paper, cultivable actinobacteria were isolated, cultured, and identified from the heavily algal-bloomed waters of Taihu Lake using 16S rRNA gene sequencing. Among the isolates, a single strain exhibiting vigorous cyanocidal activity against Microcystis aeruginosa FACHB-905 was selected for further investigation. The cyanocidal efficacy and underlying mechanisms of this strain, designated TH05, were assessed through using chlorophyll content, cyanobacterial inhibition rate, and cyanobacterial cell morphology measurements. In addition, oxidative stress responses, expression of key functional genes in FACHB-905, and variations in microcystin concentrations were comprehensively evaluated. Cyanobacterial blooms caused by Microcystis aeruginosa pose serious ecological and public health threats due to the release of microcystins (MCs). In this study, we evaluated the cyanocidal activity and mechanism of a novel actinomycete strain, Streptomyces sp. TH05. Optimization experiments revealed that a light–dark cycle of 12 h/12 h, temperature of 25 °C, and pH 7 significantly enhanced cyanocidal efficacy. Under these conditions, TH05 achieved an 84.31% inhibition rate after seven days of co-cultivation with M. aeruginosa. Scanning electron microscopy revealed two distinct cyanocidal modes: direct physical attachment of TH05 mycelia to cyanobacterial cells, causing cell wall disruption, and indirect membrane damage via extracellular bioactive compounds. Biochemical analyses showed increased levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) during the first five days, peaking at 2.47-, 2.12-, and 1.91-fold higher than control levels, respectively, indicating elevated oxidative stress. Gene expression analysis using elf-p as a reference showed that TH05 modulated key genes associated with photosynthesis (PsaB, PstD1, PstD2, RbcL), DNA repair and stress response (RecA, FtsH), and microcystin biosynthesis (McyA, McyD). All genes were upregulated except for RbcL, which was downregulated. In parallel, microcystin content peaked at 32.25 ng/L on day 1 and decreased to 16.16 ng/L by day 9, which was significantly lower than that of the control group on day 9 (29.03 ng/L). These findings suggest that strain TH05 exhibits potent and multifaceted cyanocidal activity, underscoring its potential for application in the biological control of cyanobacterial blooms. Full article

(This article belongs to the Topic Research on Natural Bioactive Product-Based Pesticidal Agents—2nd Edition)

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14 pages, 1611 KiB

Open AccessArticle

Explaining Echis: Proteotranscriptomic Profiling of Echis carinatus carinatus Venom

by Salil Javed, Prasad Gopalkrishna Gond, Arpan Samanta, Ajinkya Unawane, Muralidhar Nayak Mudavath, Anurag Jaglan and Kartik Sunagar

Toxins 2025, 17(7), 353; https://doi.org/10.3390/toxins17070353 - 16 Jul 2025

Viewed by 1035

Abstract

Snakebite remains the most neglected tropical disease globally, with India experiencing the highest rates of mortality and morbidity. While most envenomation cases in India are attributed to the ‘big four’ snakes, research has predominantly focused on Russell’s viper (Daboia russelii), [...] Read more.

Snakebite remains the most neglected tropical disease globally, with India experiencing the highest rates of mortality and morbidity. While most envenomation cases in India are attributed to the ‘big four’ snakes, research has predominantly focused on Russell’s viper (Daboia russelii), spectacled cobra (Naja naja), and common krait (Bungarus caeruleus), leading to a considerable gap in our understanding of saw-scaled viper (Echis carinatus carinatus) venoms. For instance, the venom gland transcriptome and inter- and intra-population venom variation in E. c. carinatus have largely remained uninvestigated. A single study to date has assessed the effectiveness of commercial antivenoms against this species under in vivo conditions. To address these crucial knowledge gaps, we conducted a detailed investigation of E. c. carinatus venom and reported the first venom gland transcriptome. A proteotranscriptomic evaluation revealed snake venom metalloproteinases, C-type lectins, L-amino acid oxidases, phospholipase A₂s, and snake venom serine proteases as the major toxins. Moreover, we assessed the intra-population venom variation in this species using an array of biochemical analyses. Finally, we determined the venom toxicity and the neutralising efficacy of a commercial antivenom using a murine model of snake envenoming. Our results provide a thorough molecular and functional profile of E. c. carinatus venom. Full article

(This article belongs to the Special Issue Venom Genes and Genomes of Venomous Animals: Evolution and Variation)

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19 pages, 6405 KiB

Open AccessArticle

The Venom Proteome of the Ecologically Divergent Australian Elapid, Southern Death Adder Acanthophis antarcticus

by Theo Tasoulis, C. Ruth Wang, Shaun Ellis, Tara L. Pukala, Joanna Sumner, Kate Murphy, Nathan Dunstan and Geoffrey K. Isbister

Toxins 2025, 17(7), 352; https://doi.org/10.3390/toxins17070352 - 14 Jul 2025

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Abstract

The composition of Australian snake venoms is the least well-known of any continent. We characterised the venom proteome of the southern death adder Acanthophis antarcticus—one of the world’s most morphologically and ecologically divergent elapids. Using a combined bottom-up proteomic and venom gland [...] Read more.

The composition of Australian snake venoms is the least well-known of any continent. We characterised the venom proteome of the southern death adder Acanthophis antarcticus—one of the world’s most morphologically and ecologically divergent elapids. Using a combined bottom-up proteomic and venom gland transcriptomic approach employing reverse-phase chromatographic and gel electrophoretic fractionation strategies in the bottom-up proteomic workflow, we characterised 92.8% of the venom, comprising twelve different toxin identification hits belonging to seven toxin families. The most abundant protein family was three-finger toxins (3FTxs; 59.8% whole venom), consisting mostly of one long-chain neurotoxin, alpha-elapitoxin-Aa2b making up 59% of the venom and two proteoforms of another long-chain neurotoxin. Phospholipase A₂s (PLA₂s) were the second most abundant, with four different toxins making up 22.5% of the venom. One toxin was similar to two previous non-neurotoxic PLA₂s, making up 16% of the venom. The remaining protein families present were CTL (3.6%), NGF (2.5%), CRiSP (1.8%), LAAO (1.4%), and AChE (0.8%). A. antarcticus is the first Australian elapid characterised that has a 3FTx dominant venom, a composition typical of elapids on other continents, particularly cobras Naja sp. The fact that A. antarcticus has a venom composition similar to cobra venom while having a viper-like ecology illustrates that similar venom expressions can evolve independently of ecology. The predominance of post-synaptic neurotoxins (3FTxs) and pre-synaptic neurotoxins (PLA₂) is consistent with the neurotoxic clinical effects of envenomation in humans. Full article

(This article belongs to the Section Animal Venoms)

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4 pages, 154 KiB

Open AccessEditorial

Assessing the Use of BotulinumtoxinA for Hyperactive Urinary Tract Dysfunction a Decade After Approval

by Heinrich Schulte-Baukloh

Toxins 2025, 17(7), 351; https://doi.org/10.3390/toxins17070351 - 13 Jul 2025

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Abstract

Botulinum toxin was approved by the Food and Drug Administration in 2011 for neurogenic bladder and in 2013 for idiopathic overactive bladder (OAB) [...] Full article

(This article belongs to the Special Issue Botulinum Toxin for Urinary Tract Disease: After a Decade from Approval)

15 pages, 264 KiB

Open AccessReview

Intramuscular Botulinum Toxin for Complex Regional Pain Syndrome: A Narrative Review of Published Cases

by Marc Klee, Nilkolaj la Cour Karottki and Bo Biering-Sørensen

Toxins 2025, 17(7), 350; https://doi.org/10.3390/toxins17070350 - 11 Jul 2025

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Abstract

Background: Since the 1980s, numerous case reports have explored the use of intramuscular botulinum toxin (BoNT) for Complex Regional Pain Syndrome (CRPS), with significant variation in rationale, dosing, guidance techniques, and outcome measures. This narrative review aims to summarize published evidence on the [...] Read more.

Background: Since the 1980s, numerous case reports have explored the use of intramuscular botulinum toxin (BoNT) for Complex Regional Pain Syndrome (CRPS), with significant variation in rationale, dosing, guidance techniques, and outcome measures. This narrative review aims to summarize published evidence on the use of intramuscular BoNT in patients with CRPS, including studies using earlier terminology such as reflex sympathetic dystrophy (RSD). Given the heterogeneous and largely anecdotal nature of the literature, this review is intended to map the existing landscape rather than conduct a formal analysis. Methods: The PubMed and EMBASE databases were searched in August 2024 using terms related to CRPS and botulinum toxin. Following abstract and full-text screening, 25 publications were included. Results: The included studies span single case reports, case series, and small cohorts, encompassing at least 96 individual CRPS patients treated with intramuscular BoNT. Reported outcomes were heterogeneous, and key treatment parameters—such as toxin type, target muscles, guidance technique, and dosing—were inconsistently reported. Conclusion: The evidence for intramuscular BoNT in CRPS remains limited and heterogeneous, preventing firm conclusions on its efficacy or safety. Its use may be considered in select cases, particularly those with disabling or painful focal dystonia or myofascial pain, but standardized prospective studies are needed to clarify its clinical role. Full article

(This article belongs to the Special Issue Botulinum Neurotoxins for the Treatment of Chronic Pain and Headaches)

17 pages, 10456 KiB

Open AccessArticle

Efficacy and Safety of Letibotulinum Toxin A for the Treatment of Melasma in Two Different Dilutions: A Randomized Double-Blind Split-Face Study

by Juthapa Pongklaokam, Woraphong Manuskiatti, Rungsima Wanitphakdeedecha, Pitchaya Maneeprasopchoke, Panwadee Thongjaroensirikul, Yanin Nokdhes, Rona Maria R. Abad-Constantino, Woramate Bhorntarakcharoen, Sariya Sittiwanaruk and Thanya Techapichetvanich

Toxins 2025, 17(7), 349; https://doi.org/10.3390/toxins17070349 - 11 Jul 2025

Viewed by 985

Abstract

Background: Melasma is an acquired hyperpigmentation disorder with multifactorial etiologies and limited response to conventional therapies. Recent evidence suggests that Botulinum Toxin A (BoNT-A) may modulate ultraviolet (UV)-induced pigmentation and offer therapeutic benefits. Objective: We sought to evaluate the efficacy and safety of [...] Read more.

Background: Melasma is an acquired hyperpigmentation disorder with multifactorial etiologies and limited response to conventional therapies. Recent evidence suggests that Botulinum Toxin A (BoNT-A) may modulate ultraviolet (UV)-induced pigmentation and offer therapeutic benefits. Objective: We sought to evaluate the efficacy and safety of two intradermal dilutions of Letibotulinum toxin A (LetiBoNT-A) in Thai patients with melasma. Methods: In this randomized, double-blind, split-face study, 30 participants aged 32–62 years received a single intradermal injection of LetiBoNT-A, with 20 units administered per cheek. A 1:5 dilution was injected on one side of the face, and a 1:10 dilution was injected on the contralateral side. Outcomes were evaluated over a 6-month period using the Hemi-modified Melasma Area and Severity Index (Hemi-mMASI), VISIA^® brown spot analysis, and quantitative assessments of skin texture. Results: Both dilutions significantly improved Hemi-mMASI scores (1:5, p = 0.043; 1:10, p = 0.002) and brown spots (1:5, p = 0.002; 1:10, p < 0.001). The 1:10 dilution showed earlier and more sustained improvements. Subgroup analysis revealed greater reductions in Hemi-mMASI scores among patients with telangiectatic melasma, particularly with the 1:10 dilution, though they were not statistically significant. Additionally, the 1:10 dilution significantly reduced pore volume, pore area, and sebum levels. One case of transient facial asymmetry was reported with the 1:5 dilution. Conclusions: LetiBoNT-A is a safe and effective adjunct in melasma treatment. The 1:10 dilution offered superior clinical outcomes. Full article

(This article belongs to the Special Issue The Evolving Role of Botulinum Toxin in Clinical Therapeutics)

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46 pages, 3177 KiB

Open AccessReview

Recent Advancements in Lateral Flow Assays for Food Mycotoxin Detection: A Review of Nanoparticle-Based Methods and Innovations

by Gayathree Thenuwara, Perveen Akhtar, Bilal Javed, Baljit Singh, Hugh J. Byrne and Furong Tian

Toxins 2025, 17(7), 348; https://doi.org/10.3390/toxins17070348 - 11 Jul 2025

Viewed by 626

Abstract

Mycotoxins are responsible for a multitude of diseases in both humans and animals, resulting in significant medical and economic burdens worldwide. Conventional detection methods, such as enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), and liquid chromatography-tandem mass spectrometry (LC-MS/MS), are highly effective, [...] Read more.

Mycotoxins are responsible for a multitude of diseases in both humans and animals, resulting in significant medical and economic burdens worldwide. Conventional detection methods, such as enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), and liquid chromatography-tandem mass spectrometry (LC-MS/MS), are highly effective, but they are generally confined to laboratory settings. Consequently, there is a growing demand for point-of-care testing (POCT) solutions that are rapid, sensitive, portable, and cost-effective. Lateral flow assays (LFAs) are a pivotal technology in POCT due to their simplicity, rapidity, and ease of use. This review synthesizes data from 78 peer-reviewed studies published between 2015 and 2024, evaluating advances in nanoparticle-based LFAs for detection of singular or multiplex mycotoxin types. Gold nanoparticles (AuNPs) remain the most widely used, due to their favorable optical and surface chemistry; however, significant progress has also been made with silver nanoparticles (AgNPs), magnetic nanoparticles, quantum dots (QDs), nanozymes, and hybrid nanostructures. The integration of multifunctional nanomaterials has enhanced assay sensitivity, specificity, and operational usability, with innovations including smartphone-based readers, signal amplification strategies, and supplementary technologies such as surface-enhanced Raman spectroscopy (SERS). While most singular LFAs achieved moderate sensitivity (0.001–1 ng/mL), only 6% reached ultra-sensitive detection (<0.001 ng/mL), and no significant improvement was evident over time (ρ = −0.162, p = 0.261). In contrast, multiplex assays demonstrated clear performance gains post-2022 (ρ = −0.357, p = 0.0008), largely driven by system-level optimization and advanced nanomaterials. Importantly, the type of sample matrix (e.g., cereals, dairy, feed) did not significantly influence the analytical sensitivity of singular or multiplex lateral LFAs (Kruskal–Wallis p > 0.05), confirming the matrix-independence of these optimized platforms. While analytical challenges remain for complex targets like fumonisins and deoxynivalenol (DON), ongoing innovations in signal amplification, biorecognition chemistry, and assay standardization are driving LFAs toward becoming reliable, ultra-sensitive, and field-deployable platforms for high-throughput mycotoxin screening in global food safety surveillance. Full article

(This article belongs to the Special Issue Advances in Contamination, Detection and Risk Assessment of Mycotoxins)

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15 pages, 1732 KiB

Open AccessArticle

Differentiating Zeranol Implant Abuse and Fusarium spp. Toxin-Contaminated Corn Intake by Detection and Quantification of Resorcylic Acid Lactones in Bovine Urine

by Rafael Silva Gomes, Vanessa Gonçalves dos Santos, Carlos Juliano da Silva, Amanda Martinez Nagato Simões, Eliene Alves dos Santos, Mary Ane Gonçalves Lana, Kelly Moura Keller, Marco Blokland, Ane Arrizabalaga-Larrañaga, Rafael Romero Nicolino, Marcelo Resende de Souza, Tadeu Chaves de Figueiredo, Saskia Sterk and Silvana de Vasconcelos Cançado

Toxins 2025, 17(7), 347; https://doi.org/10.3390/toxins17070347 - 11 Jul 2025

Viewed by 402

Abstract

Resorcylic acid lactones (RALs) are fungal metabolites with known biological activity. Zeranol, a synthetic RAL, has been used as an estrogenic growth promoter in cattle; however, its use is prohibited in several countries. Zearalenone, a mycotoxin produced by Fusarium spp., is commonly found [...] Read more.

Resorcylic acid lactones (RALs) are fungal metabolites with known biological activity. Zeranol, a synthetic RAL, has been used as an estrogenic growth promoter in cattle; however, its use is prohibited in several countries. Zearalenone, a mycotoxin produced by Fusarium spp., is commonly found in contaminated animal feed and can be metabolized into other RALs, which are subsequently excreted in urine. To differentiate between natural contamination from feed and the illegal administration of zeranol, the European Union Reference Laboratory for Growth Promoters (EURL) developed a mathematical equation. This study aims to evaluate the detection and quantification of RALs in bovine urine from animals fed zearalenone-contaminated diets, implanted with zeranol, or subjected to both conditions. RALs were detected and quantified in the urine of cattle consuming contaminated corn, while zeranol and taleranol were identified in the urine of implanted animals. The EURL equation proved to be a valuable tool for determining the origin of RALs in bovine urine and holds significant potential for monitoring and enforcing regulations regarding the illegal use of zeranol. Full article

(This article belongs to the Section Mycotoxins)

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17 pages, 5008 KiB

Open AccessArticle

Biodegradation of Microcystins by Aquatic Bacteria Klebsiella spp. Isolated from Lake Kasumigaura

by Thida Lin, Kazuya Shimizu, Tianxiao Liu, Qintong Li and Motoo Utsumi

Toxins 2025, 17(7), 346; https://doi.org/10.3390/toxins17070346 - 10 Jul 2025

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Abstract

Microcystins (MCs) are the most toxic and abundant cyanotoxins found in natural waters during harmful cyanobacterial blooms. These toxins pose a significant threat to plant, animal, and human health due to their toxicity. Degradation of MCs by MC-degrading bacteria is a promising method [...] Read more.

Microcystins (MCs) are the most toxic and abundant cyanotoxins found in natural waters during harmful cyanobacterial blooms. These toxins pose a significant threat to plant, animal, and human health due to their toxicity. Degradation of MCs by MC-degrading bacteria is a promising method for controlling these toxins, demonstrating safety, high efficiency, and cost-effectiveness. In this study, we isolated potential MC-degrading bacteria (strains TA13, TA14, and TA19) from Lake Kasumigaura in Japan and found that they possess a high capacity for MC degradation. Based on 16S rRNA gene sequencing, all three isolated strains were identified as belonging to the Klebsiella species. These bacteria effectively degraded MC-RR, MC-YR, and MC-LR under various temperature and pH conditions within 10 h, with the highest degrading activity and degradation rate observed at 40 °C. Furthermore, the isolated strains efficiently degraded MCs not only under neutral pH conditions, but also in alkaline environments. Additionally, we detected the MC-degrading gene (mlrA) in all three isolated strains, marking the first report of the mlrA gene in Klebsiella species. The copy number of the mlrA gene in the strains increased after exposure to MCs. These findings indicate that strains TA13, TA14, and TA19 significantly contribute of MC bioremediation in Lake Kasumigaura during cyanobacterial blooms. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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20 pages, 1188 KiB

Open AccessReview

Increased Cardiovascular Mortality in Hemodialysis: The Role of Chronic Inflammation, Complement Activation, and Non-Biocompatibility

by Ákos Géza Pethő, Tibor Fülöp, Petronella Orosz, Gábor Szénási, Mihály Tapolyai and László Dézsi

Toxins 2025, 17(7), 345; https://doi.org/10.3390/toxins17070345 - 10 Jul 2025

Viewed by 598

Abstract

Background: Chronic kidney disease (CKD) is an established global health problem, with the increased prevalence of vascular inflammation, accelerated atherogenesis, and thrombotic risk all contributing to overall cardiovascular risk. The major CKD-specific risk factor is presumed to be the accumulation of uremic toxins [...] Read more.

Background: Chronic kidney disease (CKD) is an established global health problem, with the increased prevalence of vascular inflammation, accelerated atherogenesis, and thrombotic risk all contributing to overall cardiovascular risk. The major CKD-specific risk factor is presumed to be the accumulation of uremic toxins in circulation and tissues, further accelerating the progression of CKD and its co-morbidities, including those of bone mineral disorders and cardiovascular diseases. Materials and Methods: In our narrative review, we focused on non-traditional cardiovascular risk factors, as they evolve with declined kidney function and are potentially further modulated by the choice of kidney replacement therapy. Results: Based on the data from the literature to date, the pre-eminent role of non-traditional risk factors emerges to mediate inflammation and increased cardiovascular mortality. In particular, patients receiving hemodialysis (HD) display dramatically increased CVD-mediated mortality. This intensified state of inflammation may be linked to the direct exposure of the bloodstream to a bio-incompatible environment in HD; for both complement-mediated and non-complement-mediated reactions, the possible contribution of neutrophil extracellular traps and complement activation-related pseudoallergy are reviewed in detail. Conclusions: Our narrative review emphasizes key elements of a bio-incompatible HD environment that may contribute to increased cardiovascular mortality in patients receiving HD. Summarizing these results may provide conceptual opportunities to develop new therapeutic targets. Full article

(This article belongs to the Section Uremic Toxins)

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27 pages, 768 KiB

Open AccessReview

Pre-Harvest Aflatoxin Contamination in Crops and Climate Change Factors: A European Overview

by Ainhoa Bereziartua, Anke Huss, Jannigje G. Kers, Lidwien A. M. Smit, Roel Vermeulen and Daniel Martins Figueiredo

Toxins 2025, 17(7), 344; https://doi.org/10.3390/toxins17070344 - 8 Jul 2025

Viewed by 650

Abstract

Aflatoxin (AF) contamination of crops during the pre-harvest period is a significant global concern for food and feed safety (FFS). In Europe, climate change presents a growing threat to agricultural products by increasing the risk of AF contamination. This umbrella review evaluates the [...] Read more.

Aflatoxin (AF) contamination of crops during the pre-harvest period is a significant global concern for food and feed safety (FFS). In Europe, climate change presents a growing threat to agricultural products by increasing the risk of AF contamination. This umbrella review evaluates the scope and quality of pre-harvest data on climate-related AF contamination in Europe, addressing key questions: What insights do researchers provide on the relationship between climate change and pre-harvest AF contamination, and what data are lacking? Which crops are the focus of current research, and where in Europe are these studies concentrated? How is the data presented, and is it standardized? We conducted an umbrella literature review, extracting relevant studies from PubMed and Scopus up to 14 October 2024. Our findings indicate that rising temperatures, droughts, and shifting rainfall patterns increasingly favor the growth of aflatoxigenic fungi and pre-harvest AF contamination in European crops, posing risks to FFS and agricultural stability. However, inconsistencies in data collection and reporting limit cross-regional comparisons and hinder the development of effective mitigation strategies. Standardizing methodologies and improving data accessibility will enhance predictive modeling, strengthen risk assessments, and support targeted adaptation efforts, providing actionable insights for policymakers and agricultural stakeholders. Full article

(This article belongs to the Section Mycotoxins)

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19 pages, 1475 KiB

Open AccessArticle

Chemical and Morphological Constitutive Defensive Traits of Cyanobacteria Have Different Effects on the Grazing of a Small Tropical Cladoceran

by Luciana Machado Rangel, Marcella Coelho Berjante Mesquita, Shara Rosa de Barros, Vinicius Neres-Lima, Michael Ribas Celano, Mauro Cesar Palmeira Vilar, Sandra Maria Feliciano de Oliveira e Azevedo and Marcelo Manzi Marinho

Toxins 2025, 17(7), 343; https://doi.org/10.3390/toxins17070343 - 5 Jul 2025

Viewed by 587

Abstract

Antipredator defenses of bloom-forming cyanobacteria species maximize their fitness but can reduce carbon and energy transfer efficiency to higher trophic levels, making them a key regulator of plankton communities in eutrophic waters. We investigated the grazing responses of the tropical cladoceran Moina micrura [...] Read more.

Antipredator defenses of bloom-forming cyanobacteria species maximize their fitness but can reduce carbon and energy transfer efficiency to higher trophic levels, making them a key regulator of plankton communities in eutrophic waters. We investigated the grazing responses of the tropical cladoceran Moina micrura to different strains of the cyanobacteria Microcystis aeruginosa and Planktothrix isothrix, using a good food source (green algae Mono-raphidium capricornutum) as a control. Both Microcystis strains grow as unicellular and are microcystins producers; however, this cyanotoxin was not detected on the filamentous Planktothrix strains. M. micrura ingested all cyanobacteria at reduced rates compared to single diets with Monoraphidium. In mixed diets, food type had a significant effect on grazing responses, which differed interspecifically. Planktothrix was more grazed than Microcystis strains. Feeding selectivity on Monoraphidium was negatively affected by the increase of cyanobacteria in the diet. We observed varied responses across treatments, ranging from feeding inhibition to different degrees of tolerance toward cyanobacteria, particularly in non-microcystin-producing species. We also highlight the selectivity of small tropical cladocerans, a pattern that is not yet well documented. These findings emphasize that studies incorporating phyto- and zooplankton with a history of coexistence can provide more meaningful insights into natural ecosystem dynamics. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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20 pages, 3269 KiB

Open AccessArticle

Immunomodulatory Effect of a Cysteine-Rich Secretory Protein from an Entomopathogenic Nematode with Sterol-Binding Activity

by Jorge Frias, Duarte Toubarro, Tiago Paiva and Nelson Simões

Toxins 2025, 17(7), 342; https://doi.org/10.3390/toxins17070342 - 5 Jul 2025

Viewed by 481

Abstract

The Steinernema carpocapsae nematode is known to release several excretory/secretory products (ESPs) in its venom upon contact and during the parasitic infection process of insect hosts. A recurrent family of proteins found in this nematode’s venom is the CAP (cysteine-rich secretory protein/antigen 5/pathogenesis-related [...] Read more.

The Steinernema carpocapsae nematode is known to release several excretory/secretory products (ESPs) in its venom upon contact and during the parasitic infection process of insect hosts. A recurrent family of proteins found in this nematode’s venom is the CAP (cysteine-rich secretory protein/antigen 5/pathogenesis-related 1) protein, but the functional role of these proteins remains unknown. To elucidate the biological function, this study focused on characterising the secreted protein, first identified in the venom of the nematode’s parasitic stage, and the sequence retrieved from transcriptomic analysis. The structural comparisons of the Sc-CAP protein model, as determined by AlphaFold2, revealed related structures from other parasitic nematodes of vertebrates. Some of these closely related proteins are reported to have sterol-binding ability. The Sc-CAP recombinant protein was successfully produced in Escherichia coli in conjunction with a chaperone protein. The results showed that the Sc-CAP protein binds to cholesterol, and docking analyses of sterols on the protein revealed potential molecular interactions. Immunoassays performed in Galleria mellonella larvae revealed that this venom protein has an inhibitory effect against phenoloxidase and the antimicrobial response of insects. This suggests that the venom protein has an immunomodulatory function against insects, emphasising its importance during the parasite–host interaction. Full article

(This article belongs to the Section Animal Venoms)

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15 pages, 881 KiB

Open AccessArticle

Economic Evaluation of the InTENSE Program of Therapy Alongside Botulinum Neurotoxin a for the Rehabilitation of Chronic Upper Limb Spasticity

by Rachel Milte, Jia Song, Sean Docking, Julie Ratcliffe, Ian D. Cameron, Maria Crotty, Louise Ada, Coralie English and Natasha A. Lannin

Toxins 2025, 17(7), 341; https://doi.org/10.3390/toxins17070341 - 4 Jul 2025

Viewed by 518

Abstract

Spasticity is a persistent and debilitating consequence of stroke and effective rehabilitation is a healthcare priority. Botulinum neurotoxin A (BoNT-A) with supportive therapy has increasingly been embedded within clinical practice for treatment of post-stroke spasticity. But the evidence for this approach has hitherto [...] Read more.

Spasticity is a persistent and debilitating consequence of stroke and effective rehabilitation is a healthcare priority. Botulinum neurotoxin A (BoNT-A) with supportive therapy has increasingly been embedded within clinical practice for treatment of post-stroke spasticity. But the evidence for this approach has hitherto been limited to the findings of a limited number of small trials. The InTENSE trial was undertaken specifically to provide high-quality clinical trial evidence focusing on the effect of BoNT-A and adjunctive therapy on upper limb spasticity. While the clinical trial did not detect a significant impact upon clinical outcomes, there remains a need to evaluate any impact on the broader use of healthcare resources and overall cost-effectiveness. A detailed cost–utility analysis of the InTENSE trial was undertaken. The costs over the 12-month follow-up period were compared with quality-adjusted life years (QALY) gained using utilities generated from the EQ-5D three level (EQ-5D-3L) instrument. There were no significant differences in QALY gained between the intervention and control groups identified, or in the majority of health and community care costs. The Incremental Cost-Effectiveness Ratio per QALY gained was estimated at AU $63,947.11 (Australian dollars), which is well above accepted thresholds for cost-effectiveness in Australia. The study was unable to identify evidence for the cost-effectiveness of treatment approaches combining BoNT-A with adjunctive therapy. Full article

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23 pages, 2062 KiB

Open AccessReview

Potential Compounds as Inhibitors of Staphylococcal Virulence Factors Involved in the Development of Thrombosis

by Anna Lichota, Krzysztof Gwozdzinski and Monika Sienkiewicz

Toxins 2025, 17(7), 340; https://doi.org/10.3390/toxins17070340 - 4 Jul 2025

Viewed by 415

Abstract

For many years, staphylococci have been detected mainly in infections of the skin and soft tissues, organs, bone inflammations, and generalized infections. Thromboembolic diseases have also become a serious plague of our times, which, as it turns out, are closely related to the [...] Read more.

For many years, staphylococci have been detected mainly in infections of the skin and soft tissues, organs, bone inflammations, and generalized infections. Thromboembolic diseases have also become a serious plague of our times, which, as it turns out, are closely related to the toxic effects of staphylococci. Staphylococcus aureus, because of the presence of many different kinds of virulence factors, is capable of manipulating the host’s innate and adaptive immune responses. These include toxins and cofactors that activate host zymogens and exoenzymes, as well as superantigens, which are highly inflammatory and cause leukocyte death. Coagulases and staphylokinases can control the host’s coagulation system. Nucleases and proteases inactivate various immune defense and surveillance proteins, including complement components, peptides and antibacterial proteins, and surface receptors that are important for leukocyte chemotaxis. On the other hand, secreted toxins and exoenzymes are proteins that disrupt the endothelial and epithelial barrier as a result of cell lysis and disintegration of linking proteins, which ultimately increases the risk of thromboembolism. In this review, we discuss various virulence factors and substances that may inhibit their activity. Full article

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15 pages, 4607 KiB

Open AccessArticle

Transcriptome Analysis of Dimethyl Fumarate Inhibiting the Growth of Aspergillus carbonarius

by Siruo Wang, Bowen Tai, Xifan Yu, Erfeng Li, Gang Wang, Jing Jin and Fuguo Xing

Toxins 2025, 17(7), 339; https://doi.org/10.3390/toxins17070339 - 4 Jul 2025

Viewed by 405

Abstract

Aspergillus carbonarius is one of the main pathogens responsible for postharvest diseases in fruits and is also one of the main ochratoxin A-producing strains. It not only causes significant economic losses but also poses a risk to human health. We found an inhibitory [...] Read more.

Aspergillus carbonarius is one of the main pathogens responsible for postharvest diseases in fruits and is also one of the main ochratoxin A-producing strains. It not only causes significant economic losses but also poses a risk to human health. We found an inhibitory effect of dimethyl fumarate fumigation on the growth of A. carbonarius. To further explore its antifungal mechanism, this study elucidated the functions of key pathway-related genes through a transcriptomics analysis. A total of 1402 differentially expressed genes (DEGs) were identified, including 987 up-regulated and 415 down-regulated genes. Dimethyl fumarate was found to significantly inhibit the growth of A. carbonarius by disrupting cell integrity and obstructing mycelium growth and secondary metabolism. These findings provide a basis for the potential application of dimethyl fumarate in the food industry to inhibit A. carbonarius and subsequent ochratoxin A contamination. Full article

(This article belongs to the Section Mycotoxins)

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20 pages, 10410 KiB

Open AccessArticle

Modeling Algal Toxin Dynamics and Integrated Web Framework for Lakes

by Özlem Baydaroğlu, Serhan Yeşilköy, Anchit Dave, Marc Linderman and Ibrahim Demir

Toxins 2025, 17(7), 338; https://doi.org/10.3390/toxins17070338 - 3 Jul 2025

Viewed by 524

Abstract

Harmful algal blooms (HABs) are one of the major environmental concerns, as they have various negative effects on public and environmental health, recreational services, and economics. HAB modeling is challenging due to inconsistent and insufficient data, as well as the nonlinear nature of [...] Read more.

Harmful algal blooms (HABs) are one of the major environmental concerns, as they have various negative effects on public and environmental health, recreational services, and economics. HAB modeling is challenging due to inconsistent and insufficient data, as well as the nonlinear nature of algae formation data. However, it is crucial for attaining sustainable development goals related to clean water and sanitation. From this point of view, we employed the sparse identification nonlinear dynamics (SINDy) technique to model microcystin, an algal toxin, utilizing dissolved oxygen as a water quality metric and evaporation as a meteorological parameter. SINDy is a novel approach that combines a sparse regression and machine learning method to reconstruct the analytical representation of a dynamical system. The model results indicate that MAPE values of approximately 2% were achieved in three out of four lakes, while the MAPE value of the remaining lake is 11%. Moreover, a model-driven and web-based interactive tool was created to develop environmental education, raise public awareness on HAB events, and produce more effective solutions to HAB problems through what-if scenarios. This interactive and user-friendly web platform allows tracking the status of HABs in lakes and observing the impact of specific parameters on harmful algae formation. Full article

(This article belongs to the Special Issue Managing and Preventing Harmful Algal Blooms (HABs): Do We Know Enough? (2nd Edition))

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18 pages, 3265 KiB

Open AccessArticle

Nymphoides peltata Alleviates Patulin-Induced Glutamine Metabolic Stress and Epithelial Toxicity in Small Intestinal Epithelial Cells

by Chae Hyun Lee, Sangsu Shin, Tae Hyun Kim and Sang In Lee

Toxins 2025, 17(7), 337; https://doi.org/10.3390/toxins17070337 - 3 Jul 2025

Viewed by 504

Abstract

Patulin (PAT) is a mycotoxin commonly found in fruits and contaminated feedstuffs, known for its gastrointestinal and systemic toxicity. However, the mechanisms underlying PAT-induced damage to intestinal epithelial cells remain poorly understood. In this study, we demonstrated that 6.5 µM PAT exposure for [...] Read more.

Patulin (PAT) is a mycotoxin commonly found in fruits and contaminated feedstuffs, known for its gastrointestinal and systemic toxicity. However, the mechanisms underlying PAT-induced damage to intestinal epithelial cells remain poorly understood. In this study, we demonstrated that 6.5 µM PAT exposure for 24 h reduced glutamine (GLN) uptake and altered the expression of GLN transporters and related metabolic enzymes in IPEC-J2 cells. This concentration was selected based on previous in vitro studies that reported PAT-induced cytotoxicity in porcine intestinal epithelial cells. Moreover, PAT also upregulated ER stress markers (DDIT3, EIF2AK3, ERN1, and HSPA5) and inflammatory cytokines (IL-8, IL-1β, and TNF-α), while decreasing ZO-1 localization, indicating disrupted epithelial barrier integrity. Although 6 mM GLN supplementation only partially mediated ER stress and inflammatory responses, it more effectively restored ZO-1 localization. A high-throughput screening of 324 natural products was conducted to identify potential protective agents, identifying Nymphoides peltata extract as a promising candidate. Co-treatment with 80 ng/μL N. peltata extract improved GLN uptake, partially alleviated ER stress and inflammation, and significantly restored tight junction structure in PAT-exposed cells. Collectively, these findings suggest that N. peltata could serve as a novel natural therapeutic for enhancing intestinal resilience against PAT-induced toxicity. Specifically, this study highlights the potential use of N. peltata extract as a natural feed additive to protect intestinal health in livestock under mycotoxin stress. Full article

(This article belongs to the Section Mycotoxins)

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14 pages, 1948 KiB

Open AccessArticle

Establishing a 3D Spheroid Model of Cholinergic Neurons from SH-SY5Y Cells for Neurotoxicity Assessment

by Felipe Franco-Campos, Mónica Fernández-Franzón, Yelko Rodríguez-Carrasco and María-José Ruiz

Toxins 2025, 17(7), 336; https://doi.org/10.3390/toxins17070336 - 2 Jul 2025

Viewed by 553

Abstract

The nervous system maintains homeostasis and coordinated behavior through complex neuronal and glial cells. Traditional models, such as primary rodent neurons and human-induced pluripotent stem cell (hIPSC)-derived neurons, have advanced our understanding of neuronal function and neurotoxic damage; however, they are costly and [...] Read more.

The nervous system maintains homeostasis and coordinated behavior through complex neuronal and glial cells. Traditional models, such as primary rodent neurons and human-induced pluripotent stem cell (hIPSC)-derived neurons, have advanced our understanding of neuronal function and neurotoxic damage; however, they are costly and labor-intensive. SH-SY5Y cells, an immortalized human neuroblastoma cell line, provide a more accessible alternative for studying neuronal processes and neurotoxicity. However, their limited capacity to differentiate into specific neuronal phenotypes remains a challenge. To address this limitation, differentiation protocols using neuronal factors and vitamins have been developed, primarily in two-dimensional (2D) cultures, which reduces physiological relevance. Here, we present a novel three-dimensional (3D) SH-SY5Y model incorporating 2D differentiation protocols to generate cholinergic neurons (ChAT+). This model enhances neurotoxicity studies related to pesticides and mycotoxins. Our protocol produces homogeneous spheroids differentiated into cholinergic neurons using serum restriction and specific factors, maintaining viability and circularity for up to 22 days. Differentiation was validated by immunofluorescence and Western blot by Choline acetyltransferase (ChAT) expression. This scalable and reproducible 3D model provides a valuable in vitro tool for neurotoxicological research, improving physiological relevance and enabling the study of cholinergic neuron differentiation and function. Full article

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

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25 pages, 3047 KiB

Open AccessArticle

Fate of Pyrrolizidine Alkaloids in Soil: Insights from Myosotis arvensis L. and Senecio vulgaris L.

by Ilva Nakurte, Gundars Skudriņš and Ieva Mežaka

Toxins 2025, 17(7), 335; https://doi.org/10.3390/toxins17070335 - 2 Jul 2025

Viewed by 413

Abstract

Pyrrolizidine alkaloids are plant-derived toxins with environmental persistence and the potential to contaminate soil, water, and adjacent crops. This study investigated the leaching behavior and environmental fate of PAs from two PA-producing weeds—Myosotis arvensis L. (Boraginaceae) and Senecio vulgaris L. (Asteraceae)—in two [...] Read more.

Pyrrolizidine alkaloids are plant-derived toxins with environmental persistence and the potential to contaminate soil, water, and adjacent crops. This study investigated the leaching behavior and environmental fate of PAs from two PA-producing weeds—Myosotis arvensis L. (Boraginaceae) and Senecio vulgaris L. (Asteraceae)—in two Latvian agricultural soils: sandy loam and loam. Hot- and cold-water plant extracts were applied to soil columns (10 cm and 20 cm), and leachates were analyzed over a 14-day period using QuEChERS purification and LC-HRMS detection. Leaching varied by plant species, extract type, and soil. M. arvensis showed significantly higher cumulative leaching (77–84% for cold, 65–71% for hot extracts), attributed to the higher solubility of N-oxides. In contrast, S. vulgaris extracts leached minimally (<0.84% from sandy loam) and were undetectable in loam. The presence of cyclic diester PAs in S. vulgaris and the higher cation exchange capacity of loam favored retention or degradation. PANO-to-PA conversion occurred in both soils, indicating redox activity. The fate of PAs was influenced by structural type (diesters showing higher persistence), extraction method (hot extraction releasing more pyrrolizidine alkaloids), and soil properties such as pH, organic matter, and cation exchange capacity, which affected sorption and mobility. These findings underscore the significance of soil composition in controlling PA mobility and associated environmental risks. Future research should focus on long-term PA persistence across diverse soil types and investigate crop uptake potential and microbial degradation pathways under field conditions. Full article

(This article belongs to the Special Issue Toxic Plant-Derived Metabolites)

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19 pages, 2150 KiB

Open AccessArticle

Associations Between Uraemic Toxins and Gut Microbiota in Adults Initiating Peritoneal Dialysis

by Philippa James, Jordan Stanford, Ojas V. A. Dixit, Mary Ann Nicdao, Brett McWhinney, Kamal Sud, Michele Ryan, Scott Read, Golo Ahlenstiel, Kelly Lambert, Claire O’Brien and Katrina Chau

Toxins 2025, 17(7), 334; https://doi.org/10.3390/toxins17070334 - 1 Jul 2025

Viewed by 475

Abstract

Declining kidney function contributes to the accumulation of uraemic toxins produced by gut microbiota, leading to the uraemic syndrome. This study aimed to identify associations between uraemic toxins, diet quality, symptoms and the gut microbiota in individuals initiating peritoneal dialysis. A cross-sectional analysis [...] Read more.

Declining kidney function contributes to the accumulation of uraemic toxins produced by gut microbiota, leading to the uraemic syndrome. This study aimed to identify associations between uraemic toxins, diet quality, symptoms and the gut microbiota in individuals initiating peritoneal dialysis. A cross-sectional analysis of baseline data from participants in a longitudinal study was conducted. Symptom scores using the Integrated Palliative Care Outcomes Scale-Renal were recorded. Plasma p-Cresyl sulfate, indoxyl sulfate and trimethylamine N-oxide were measured using liquid chromatography-mass spectrometry. Gut microbiota was determined using 16S rRNA sequencing. Multivariate linear models examined associations across the cohort. Data from 43 participants (mean age 61 ± 13 years; 70% male; median eGFR 7 mL/min/1.73 m²) were analysed. Diabetes was the primary cause of kidney disease (51.2%). Patients were classified into ‘high’ (n = 18) and ‘low’ (n = 26) uraemic toxin groups using K-means clustering. The ‘high’ group had a lower eGFR (p < 0.05) but no differences in diet quality or symptom scores. Significant differences in alpha and beta diversity were observed between the groups (p = 0.01). The ‘high’ group had increased Catenibacterium, Prevotella, Clostridia, and decreased Ruminococcus gnavus abundances. Multivariate models identified 32 genera associated with uraemic toxins, including positive associations of Oscillospiraceae UCG-002 and UCG-005 with p-cresyl sulfate, and negative associations with Actinomyces and Enterococcus. Patients with kidney failure initiating peritoneal dialysis have distinct uraemic toxin profiles, associated with differences in microbial diversity. This phenotype was also associated with differences in residual kidney function but not with diet or symptom severity. Longitudinal studies are required to determine causality and guide therapeutic interventions. Full article

(This article belongs to the Section Uremic Toxins)

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17 pages, 1034 KiB

Open AccessArticle

Monitoring of Vitamin C Plasma Levels in a Reversible Model of Malabsorption Generated in Mice by Ebulin-f

by Daniel Arranz-Paraiso, M. Angeles Rojo, Cristina Martin-Sabroso, Manuel Cordoba-Diaz, Tomás Girbés, Manuel Garrosa and Damian Cordoba-Diaz

Toxins 2025, 17(7), 333; https://doi.org/10.3390/toxins17070333 - 30 Jun 2025

Viewed by 589

Abstract

The development of reversible animal models for the study of intestinal pathologies is essential to reduce the number of animals used in research and to better understand disease mechanisms. In this study, we present a reversible model of intestinal malabsorption through the administration [...] Read more.

The development of reversible animal models for the study of intestinal pathologies is essential to reduce the number of animals used in research and to better understand disease mechanisms. In this study, we present a reversible model of intestinal malabsorption through the administration of sublethal doses of ebulin-f, a ribosome-inactivating protein, and validate its usefulness by monitoring vitamin C absorption. The scientific community increasingly recognizes the importance of rationalizing experimental designs, optimizing treatment protocols, and minimizing the use of animals in research models. Thus, new methodologies are needed to minimize invasive sampling and to develop reversible animal models that recover physiologically post-study. Such models are essential for in vivo studies of human pathologies. Sublethal doses of ebulin-f (2.5 mg/kg) administered intraperitoneally to female Swiss CD1 mice (n = 6 per group) can cause reversible intestinal alterations in the small intestine, which offer the possibility of having a valuable reversible study model of malabsorption for the investigation of this syndrome. To verify whether nutrient absorption is altered, we used vitamin C as a traceable nutrient that can be quantified in the blood. Peripheral blood samples were collected through the retro-orbital area at 30, 80, 120, 180, and 1440 min post-administration, treated with DTT and MPA, and analyzed using a validated UV/Vis–HPLC method to indirectly determine vitamin C absorption by enterocytes. Pharmacokinetic analysis revealed significantly increased vitamin C absorption on days 1 and 3 post-treatment (AUC values of 3.65 × 10⁴ and 7.10 × 10⁴, respectively) compared to control (0.94 × 10⁴), with partial recovery by day 22 (3.27 × 10⁴). Blood concentration profiles indicate that intestinal damage peaks at day 3, followed by significant regeneration by day 22, establishing this as a viable reversible model for inflammatory bowel disease research. Full article

(This article belongs to the Special Issue Ribosome-Inactivating Proteins: From Plant Toxins to Drug Applications)

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16 pages, 5492 KiB

Open AccessArticle

Tityus serrulatus Scorpion Venom-Induced Nociceptive Responses Depend on TRPV1, Immune Cells, and Pro-Inflammatory Cytokines

by Camila R. Ferraz, Marília F. Manchope, Mariana M. Bertozzi, Telma Saraiva-Santos, Ketlem C. Andrade, Anelise Franciosi, Tiago H. Zaninelli, Julia Bagatim-Souza, Sergio M. Borghi, Denise M. Cândido, Thiago M. Cunha, Rubia Casagrande, Fábio H. Kwasniewski and Waldiceu A. Verri

Toxins 2025, 17(7), 332; https://doi.org/10.3390/toxins17070332 - 30 Jun 2025

Viewed by 510

Abstract

For centuries, researchers have been fascinated by the composition of scorpion venom and its local and systemic effects on humans. During a sting, scorpions inject peptides and proteins that can affect immune cells and neurons. While the immune and nervous systems have been [...] Read more.

For centuries, researchers have been fascinated by the composition of scorpion venom and its local and systemic effects on humans. During a sting, scorpions inject peptides and proteins that can affect immune cells and neurons. While the immune and nervous systems have been studied independently in the context of scorpion stings, here we reveal part of the mechanism by which Tityus serrulatus venom induces hyperalgesia in mice. Through behavioral, immune, imaging assays, and mice genetics, we demonstrate evidence of neuroimmune crosstalk during scorpion stings. Tityus serrulatus venom induced mechanical and thermal hyperalgesia in a dose-dependent manner, as well as overt pain-like behavior. The venom directly activated dorsal root ganglia neurons and increased the recruitment of macrophages and neutrophils, releasing pro-inflammatory cytokines TNF-α and IL-1β. Blocking TRPV1⁺ neurons, TNF-α, IL-1β, and NFκB reduced the mechanical and thermal hyperalgesia, overt pain-like behavior, and the migration of macrophages and neutrophils induced by Tityus serrulatus venom. Collectively, Tityus serrulatus venom targets primary afferent nociceptive TRPV1⁺ neurons to induce hyperalgesia through the recruitment of macrophages and neutrophils and the release of pro-inflammatory cytokines. Full article

(This article belongs to the Section Animal Venoms)

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27 pages, 2201 KiB

Open AccessReview

Toxicity, Mitigation, and Chemical Analysis of Aflatoxins and Other Toxic Metabolites Produced by Aspergillus: A Comprehensive Review

by Habtamu Fekadu Gemede

Toxins 2025, 17(7), 331; https://doi.org/10.3390/toxins17070331 - 30 Jun 2025

Viewed by 1460

Abstract

Aflatoxins, toxic secondary metabolites produced primarily by Aspergillus flavus and Aspergillus parasiticus, pose significant risks to food safety, public health, and global trade. These mycotoxins contaminate staple crops such as maize and peanuts, particularly in warm and humid regions, leading to economic [...] Read more.

Aflatoxins, toxic secondary metabolites produced primarily by Aspergillus flavus and Aspergillus parasiticus, pose significant risks to food safety, public health, and global trade. These mycotoxins contaminate staple crops such as maize and peanuts, particularly in warm and humid regions, leading to economic losses and severe health effects, including hepatocellular carcinoma, immune suppression, and growth impairment. In addition to aflatoxins, Aspergillus species produce other toxic metabolites such as ochratoxin A, sterigmatocystin, and cyclopiazonic acid, which are associated with nephrotoxic, carcinogenic, and neurotoxic effects, respectively. This review provides a comprehensive analysis of aflatoxin toxicity, mitigation strategies, and chemical detection methods. The toxicity of aflatoxins is discussed in relation to their biochemical mechanisms, carcinogenicity, and synergistic effects with other mycotoxins. Various mitigation approaches, including pre-harvest biocontrol, post-harvest storage management, and novel detoxification methods such as enzymatic degradation and nanotechnology-based interventions, are evaluated. Furthermore, advances in aflatoxin detection, including chromatographic, immunoassay, and biosensor-based methods, are explored to improve regulatory compliance and food safety monitoring. This review underscores the need for integrated management strategies and global collaboration to reduce aflatoxin contamination and its associated health and economic burdens. Future research directions should focus on genetic engineering for resistant crop varieties, climate adaptation strategies, and improved risk assessment models. Full article

(This article belongs to the Special Issue Toxicity, Mitigation and Chemical Analysis of Aflatoxins and Other Toxic Metabolites Produced by Aspergillus)

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14 pages, 368 KiB

Open AccessArticle

Long-Term Effectiveness of Onabotulinum Toxin-A in a Combined Total Endoscopic Management of Pediatric Vesicoureteral Reflux in Neurogenic Bladder Dysfunction

by Claudio Paratore, Chiara Pellegrino, Noemi Deanesi, Rebecca Pulvirenti, Maria Luisa Capitanucci and Giovanni Mosiello

Toxins 2025, 17(7), 330; https://doi.org/10.3390/toxins17070330 - 29 Jun 2025

Viewed by 362

Abstract

Vesicoureteral reflux (VUR) management in children with neurogenic bladder dysfunction (NBD) remains a clinical challenge. Total endoscopic management (TEM), combining intradetrusor Onabotulinum Toxin-A (BTX-A) and subureteric dextranomer/hyaluronic acid (Deflux^(R)) injection, offers a minimally invasive alternative. The aim of this retrospective study [...] Read more.

Vesicoureteral reflux (VUR) management in children with neurogenic bladder dysfunction (NBD) remains a clinical challenge. Total endoscopic management (TEM), combining intradetrusor Onabotulinum Toxin-A (BTX-A) and subureteric dextranomer/hyaluronic acid (Deflux^(R)) injection, offers a minimally invasive alternative. The aim of this retrospective study is to evaluate the long-term effectiveness of TEM. Inclusion criteria: symptomatic II–V grade VUR (also I in bilateral VUR) in NBD children with follow-up ≥12 months. Nineteen patients were enrolled, 24 ureters (grade I–II: 2, grade III–V: 22); 5 patients (20.8%) had bilateral VUR. Mean age at surgery: 7.6 years (1.3–17). No complications were reported. TEM was effective in 11 patients (57.9%), 3/11 requiring a second TEM treatment. VUR resolution appeared in 14 ureters (58.3%), downgrading in 6 (42.9%), persistence in 4 (28.6%). Among non-responders’ patients (8/19, 42.1%), five (26.3%) required bladder augmentation (one combined with ureteral reimplantation), one (5.3%) underwent reimplantation, and two (10.5%) continued conservative management. At bladder biopsy, 11 patients (57.9%) had chronic inflammation, 8 (42.1%) showed fibrosis; no difference in success rate was recorded. All responders required repeated BTX-A injections. Mean follow-up: 3.2 years (range 1–4.7). In selected patients, TEM appears to be a safe and effective strategy, potentially delaying or avoiding major reconstructive surgery. Full article

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Toxins, Volume 17, Issue 7 (July 2025) – 48 articles

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