Toxins

19 pages, 1666 KB

Open AccessArticle

Self-Sufficient Aflatoxin Decontamination System: MOF-Based Composite Membrane with Peroxidase-Mimic and Controlled H₂O₂ Generation

by Xiaofei Cheng, Wenzhong Zhu, Xueting Zhu, Jinmin Zhang, Jia Yang, Huali Wang, Xiaoqin Mo, Chi Zhang and Lina Wu

Toxins 2025, 17(10), 516; https://doi.org/10.3390/toxins17100516 (registering DOI) - 20 Oct 2025

Abstract

Aflatoxin B1 (AFB1) and its metabolite aflatoxin M1 (AFM1) are stable and carcinogenic mycotoxins that are commonly found in dairy products, posing serious food safety concerns. However, conventional degradation methods face limited degradation efficiency and high energy demand. Here, we develop an innovative [...] Read more.

Aflatoxin B1 (AFB1) and its metabolite aflatoxin M1 (AFM1) are stable and carcinogenic mycotoxins that are commonly found in dairy products, posing serious food safety concerns. However, conventional degradation methods face limited degradation efficiency and high energy demand. Here, we develop an innovative polyvinylidene fluoride (PVDF) composite membrane incorporating Fe/Co-based metal-organic frameworks (MOF) (Named Fe/Co-MIL-88B(NH₂)) and CaO₂ for targeted aflatoxin removal from milk. This system integrates two synergistic mechanisms: (1) hierarchical porous MOF structures enabling superior aflatoxin adsorption capacity and peroxidase-like catalytic activity, and (2) CaO₂ acts as a controllable-release H₂O₂ donor, supplying a steady flux of reactive oxygen species without the addition of exogenous H₂O₂. Moreover, the PVDF membrane with mechanical stability offers uniform immobilization of active components, which prevents the aggregation of nanozymes. As a result, the integrated membrane achieves high degradation efficiency for AFB1 and AFM1, exceeding 95% within 60 min. By eliminating external oxidant addition and minimizing collateral nutrient damage, the technology demonstrates remarkable operational stability (>10 cycles) and milk quality preservation capability. This breakthrough establishes an efficient and reusable detoxification method, providing new opportunities for mycotoxin mitigation in dairy products through spatiotemporal control of reactive oxygen species. Full article

(This article belongs to the Special Issue Detection, Biosynthesis and Control of Mycotoxins (4th Edition))

24 pages, 2023 KB

Open AccessReview

New Insights into Mycotoxin Contamination, Detection, and Mitigation in Food and Feed Systems

by Marija Kovač Tomas and Iva Jurčević Šangut

Toxins 2025, 17(10), 515; https://doi.org/10.3390/toxins17100515 - 20 Oct 2025

Abstract

Mycotoxins are ubiquitous and unavoidable contaminants in food and feed, posing significant health risks through toxicity syndromes collectively referred to as mycotoxicoses. With climate change enhancing the conditions favorable for fungal growth and mycotoxin production, concerns over food and feed safety are increasingly [...] Read more.

Mycotoxins are ubiquitous and unavoidable contaminants in food and feed, posing significant health risks through toxicity syndromes collectively referred to as mycotoxicoses. With climate change enhancing the conditions favorable for fungal growth and mycotoxin production, concerns over food and feed safety are increasingly pressing. Although regulatory frameworks have been established to monitor and limit the exposure, effective mitigation remains a challenge. This paper provides a comprehensive overview of recent advances in the prevention, detection, and control of mycotoxins, with particular emphasis on innovative strategies such as plant-derived bioactives, nanotechnology-based systems, genetic engineering approaches, antibody-mediated technology, and emerging non-thermal processing methods. Full article

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

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22 pages, 4076 KB

Open AccessArticle

Protective Effect of Ethoxyquin and N-acetylcysteine on Biochemical and Pathological Changes Induced by Chronic Exposure to Aflatoxins in Laying Hens

by María Carolina de-Luna-López, Arturo Gerardo Valdivia-Flores, Teódulo Quezada-Tristán, Raúl Ortiz-Martínez, Erika Janet Rangel-Muñoz, Emmanuel Hernández-Valdivia, Esther Albarrán-Rodríguez and Elizabeth de Santiago-Díaz

Toxins 2025, 17(10), 514; https://doi.org/10.3390/toxins17100514 - 20 Oct 2025

Abstract

Aflatoxins (AFs) represent a major threat to poultry health and food safety due to their hepatotoxic, immunosuppressive, and carcinogenic effects. This study evaluated the chemoprotective potential of ethoxyquin (EQ) and N-acetylcysteine (NAC) in laying hens (500 and 800 mg/kg of feed) exposed to [...] Read more.

Aflatoxins (AFs) represent a major threat to poultry health and food safety due to their hepatotoxic, immunosuppressive, and carcinogenic effects. This study evaluated the chemoprotective potential of ethoxyquin (EQ) and N-acetylcysteine (NAC) in laying hens (500 and 800 mg/kg of feed) exposed to chronic dietary AFs contamination (0.0–1.5 mg/kg). A total of 360 Hy-Line W36 Leghorn hens were monitored over 72 weeks using biochemical and histopathological analyses of liver and kidney tissues. NAC significantly (p < 0.01) increased hepatic and renal levels of reduced glutathione (GSH) and stimulated glutathione S-transferases (GST) and gamma-glutamyl transferase (GGT) activity, enhancing detoxification. Both agents significantly (p < 0.05) reduced plasma ALT and AST levels, preserved total protein concentrations, and attenuated liver and kidney hypertrophy. EQ demonstrated antioxidant effects, stabilizing enzymatic responses and limiting tissue damage. Histopathological analysis revealed fewer structural alterations and cellular degeneration, especially in the NAC-treated group (p < 0.01). These results suggest that NAC and EQ activate endogenous detoxification mechanisms, both enzymatic and non-enzymatic, effectively mitigating chronic aflatoxin toxicity. Their dietary supplementation offers a safe and sustainable chemoprotection strategy to support poultry health and productivity, particularly in regions facing high mycotoxin exposure. Full article

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

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29 pages, 982 KB

Open AccessReview

The Ambivalent Nature of Bacteroides fragilis and the Interaction with Clostridioides difficile: Benefits and Disadvantages for the Human Host

by Patrizia Spigaglia

Toxins 2025, 17(10), 513; https://doi.org/10.3390/toxins17100513 - 19 Oct 2025

Abstract

Bacteroides fragilis is a usually beneficial colonizer of the human gut that can also act as an opportunistic pathogen, causing infection and contributing to the development and progression of important diseases. The production and secretion of the B. fragilis toxin (BFT), the main [...] Read more.

Bacteroides fragilis is a usually beneficial colonizer of the human gut that can also act as an opportunistic pathogen, causing infection and contributing to the development and progression of important diseases. The production and secretion of the B. fragilis toxin (BFT), the main virulence factor of this bacterium, distinguishes enterotoxigenic (ETBF) from non-toxigenic (NTBF) strains. Although NTBF does not produce the BFT, certain strains can exhibit unexpected pathogenic characteristics. The complex interactions between B. fragilis and the other intestinal bacteria, such as Clostridioides difficile, the leading cause of antibiotic-associated diarrhea in healthcare settings, highlights its ambivalent role of benefactor and pathogen. In fact, although B. fragilis plays a part in preventing colonization and infection due to C. difficile (CDI), both these anaerobic bacteria can contribute to the development and progression of colorectal cancer (CRC), one of the most prevalent malignant tumors of the digestive tract. This review provides an overview of the dual nature of B. fragilis, focusing on the peculiarities of ETBF and NTBF, delving into B. fragilis interaction with C. difficile and impacts on the host. Full article

(This article belongs to the Special Issue Enterotoxigenic Bacteroides fragilis Toxin: Biological Properties and Pathogenesis)

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15 pages, 1219 KB

Open AccessArticle

Mechanisms of Cellular Responses of the Natural Alkaloid Caulerpin and Its Similarities with the Lipid-Lowering Agent Fenofibrate in Mytilus galloprovincialis

by Michela Panni, Marica Mezzelani, Maria Elisa Giuliani, Paola Nisi Cerioni, Alessandro Nardi, Ernesto Mollo, Francesco Regoli, Maura Benedetti and Stefania Gorbi

Toxins 2025, 17(10), 512; https://doi.org/10.3390/toxins17100512 - 18 Oct 2025

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Abstract

Marine-derived natural products have attracted increasing attention due to their promising pharmacological potential against various diseases. The present study investigated the hypolipidemic properties of the alkaloid caulerpin, a secondary metabolite of green algae of the genus Caulerpa, through an ex vivo approach [...] Read more.

Marine-derived natural products have attracted increasing attention due to their promising pharmacological potential against various diseases. The present study investigated the hypolipidemic properties of the alkaloid caulerpin, a secondary metabolite of green algae of the genus Caulerpa, through an ex vivo approach with Precision-Cut Tissue Slices (PCTSs) of Mytilus galloprovincialis digestive glands. PCTSs were exposed to caulerpin (CAU) (100 µM) and fenofibrate (FFB) (100 µM) for 24, 48, and 72 h. Analyses of molecular and cellular responses pertaining to lipid metabolism suggested a similar mechanism of action between CAU and FFB in marine invertebrate species, resulting in a decrease in neutral lipid content ranging from 50 to 70%. CAU’s hypolipidemic action was not associated with increased prooxidant conditions, and slower metabolism of the natural alkaloid compared to FFB was indicated by the response of biotransformation and excretion pathways. Overall, these findings provide initial insights into the potential valorization of CAU for pharmaceutical and nutraceutical applications, highlighting the need for further investigation into its mechanisms of action, possible side effects, and interactions with other drugs. Full article

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20 pages, 7890 KB

Open AccessArticle

One Bloom Is Not Like the Other—Distinct Environmental Drivers Result in Domoic Acid Events in Monterey Bay, California

by Aubrey Trapp, Andrew Baker, Kendra Hayashi and Raphael M. Kudela

Toxins 2025, 17(10), 511; https://doi.org/10.3390/toxins17100511 - 17 Oct 2025

Viewed by 215

Abstract

Domoic acid (DA), produced by Pseudo-nitzschia diatoms, is the one of the major toxin threats from harmful algal blooms (HABs) on the west coast of the United States. DA events vary in magnitude, timing, and duration, and elucidating drivers for individual events is [...] Read more.

Domoic acid (DA), produced by Pseudo-nitzschia diatoms, is the one of the major toxin threats from harmful algal blooms (HABs) on the west coast of the United States. DA events vary in magnitude, timing, and duration, and elucidating drivers for individual events is a persistent challenge. Monterey Bay experiences near-annual DA events and hosts long-term HAB monitoring at the Santa Cruz Municipal Wharf (SCW). Here we characterize two toxin events, occurring in May 2023 and March 2024. The events were similar in magnitude and duration, but an exploration of physical, biological, and chemical dynamics revealed distinct environmental drivers. These differences resulted in a significant deviation in cellular DA (cDA) within the same species of Pseudo-nitzschia. In addition, opportunistic solid-phase adsorption toxin tracking (SPATT) was used for environmental metabolomics. The novel application of SPATT revealed 159 metabolites that were strongly correlated with DA in both events and produced a spectral match to a new marine natural product using Global Natural Products Social Molecular Networking (GNPS). This work takes a multivariable approach to understanding toxin drivers and lends proof of concept for the integration of environmental metabolomics in HAB monitoring. Full article

(This article belongs to the Special Issue Ecology and Evolution of Harmful Algal Blooms)

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13 pages, 1200 KB

Open AccessArticle

Development of Lab-on-a-Chip LAMP and Real-Time PCR Assays to Detect Aflatoxigenic Aspergillus flavus and Aspergillus parasiticus in Hazelnuts

by Slavica Matić, Livio Cognolato, Martina Sanna, Monica Mezzalama, Riccardo Laurenti and Davide Spadaro

Toxins 2025, 17(10), 510; https://doi.org/10.3390/toxins17100510 - 17 Oct 2025

Viewed by 172

Abstract

Aflatoxins, which are potentially genotoxic and carcinogenic substances, are mainly produced by the Aspergillus section Flavi, including Aspergillus flavus and A. parasiticus. Current Aspergillus spp. detection is often based on molecular methods, such as real-time PCR and loop-mediated isothermal amplification (LAMP), [...] Read more.

Aflatoxins, which are potentially genotoxic and carcinogenic substances, are mainly produced by the Aspergillus section Flavi, including Aspergillus flavus and A. parasiticus. Current Aspergillus spp. detection is often based on molecular methods, such as real-time PCR and loop-mediated isothermal amplification (LAMP), targeting genes of the aflatoxin biosynthetic cluster. In this study, we developed a Lab-on-a-Chip (LoC) method based on real-time PCR and on LAMP for the specific detection of aflatoxigenic strains of A. flavus and A. parasiticus from infected hazelnuts. LoC-LAMP and LoC-real-time PCR assays were tested in terms of specificity, sensitivity, speed, and repeatability. The microfluidic chip allowed quick, specific, sensitive, simple, automatized, cheap, and user-friendly detection of aflatoxigenic strains of A. flavus and A. parasiticus. The LoC-LAMP showed a limit of detection (LOD) of 10 fg of DNA, while the LoC-real-time PCR showed a LOD of 10 pg of DNA. Achieving comparable sensitivity to that of LAMP and real-time PCR techniques, both LoC methods developed in this work offer the advantages of automation, minimal sample requirements, reagent requirements, and cost-effectiveness. Overall, the developed methods open the perspective for alternative monitoring of aflatoxigenic fungi in the agri-food industry. Full article

(This article belongs to the Section Mycotoxins)

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15 pages, 18289 KB

Open AccessArticle

Mapping Dural and Periosteal SV2C, a Botulinum Toxin A Receptor, in the Mouse

by Anisa Dehghani, Agustin Melo-Carrillo, Andrew M. Strassman, Ron S. Broide, Aubrey Manack Adams, Brett Dabruzzo, Mitchell F. Brin and Rami Burstein

Toxins 2025, 17(10), 509; https://doi.org/10.3390/toxins17100509 - 17 Oct 2025

Viewed by 217

Abstract

Objectives: There has been a long-standing debate over the presence or absence of receptors for botulinum toxin A (BoNT/A) in cephalic areas relevant to migraine pathophysiology and onabotulinumtoxinA (onabotA) sites of action in migraine prevention. To address this issue, we sought to investigate [...] Read more.

Objectives: There has been a long-standing debate over the presence or absence of receptors for botulinum toxin A (BoNT/A) in cephalic areas relevant to migraine pathophysiology and onabotulinumtoxinA (onabotA) sites of action in migraine prevention. To address this issue, we sought to investigate for the first time whether synaptic vesicle protein 2C (SV2C), one member of the SV2 receptor family, is present in axons innervating the dura and periosteum. Methods: Single- and double- labeling immunohistochemical techniques were used to map and characterize the distribution of axons containing SV2C, the third isoform of the SV2 glycoprotein, in the mouse dura and periosteum. Results: Dense networks of axons containing SV2C receptors were distributed throughout all regions of the dura and periosteum. In the dura, SV2C-LIR axons were found in 43% of all peripherin-LIR fibers, 49% of all CGRP-LIR fibers, and 75% of all NaV1.8-LIR fibers. In the periosteum, SV2C-LIR was found in 38% of all peripherin-LIR fibers, 53% of all CGRP-LIR fibers, and 68% of all NaV1.8-LIR fibers. Conclusions: We interpret these findings as suggesting that many of the labeled axons are peripheral nerve axons (peripherin-positive) of unmyelinated sensory and possibly parasympathetic origin (CGRP-positive), and that some of these sensory axons are nociceptors (NaV1.8-positive). Clinically, these findings demonstrate an abundance of axons containing onabotA receptors in the vicinity of scalp structures commonly injected with onabotA for the treatment of chronic migraine. Dense labeling in the periosteum provides another rationale for the possibility that onabotA injections in this layer of the scalp may be advantageous. Full article

(This article belongs to the Section Bacterial Toxins)

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24 pages, 3732 KB

Open AccessReview

The Elias University Hospital Approach: A Visual Guide to Ultrasound-Guided Botulinum Toxin Injection in Spasticity, Part IV—Distal Lower Limb Muscles

by Marius Nicolae Popescu, Claudiu Căpeț, Cristina Popescu and Mihai Berteanu

Toxins 2025, 17(10), 508; https://doi.org/10.3390/toxins17100508 - 16 Oct 2025

Viewed by 258

Abstract

Spasticity of the distal lower limb substantially impairs stance, gait, and quality of life in patients with upper motor neuron lesions. Although ultrasound-guided botulinum toxin A (BoNT-A) injections are increasingly employed, structured, muscle-specific visual guidance for the distal lower limb remains limited. This [...] Read more.

Spasticity of the distal lower limb substantially impairs stance, gait, and quality of life in patients with upper motor neuron lesions. Although ultrasound-guided botulinum toxin A (BoNT-A) injections are increasingly employed, structured, muscle-specific visual guidance for the distal lower limb remains limited. This study provides a comprehensive guide for ultrasound-guided BoNT-A injections across ten key distal lower limb muscles: gastrocnemius, soleus, tibialis posterior, flexor hallucis longus, flexor digitorum longus, tibialis anterior, extensor hallucis longus, flexor digitorum brevis, flexor hallucis brevis, and extensor digitorum longus. For each muscle, we present (1) Anatomical positioning relative to osseous landmarks; (2) Sonographic identification cues and dynamic features; (3) Zones of intramuscular neural arborization optimal for injection; (4) Practical injection protocols derived from literature and clinical experience. High-resolution ultrasound images and dynamic videos illustrate real-life muscle behavior and guide injection site selection. This guide facilitates precise targeting by correlating sonographic signs with optimal injection zones, addresses common spastic patterns—including equinus, varus, claw toe, and hallux deformities—and integrates fascial anatomy with motor-point mapping. This article completes the Elias University Hospital visual series, providing clinicians with a unified framework for effective spasticity management to improve gait, posture, and patient autonomy. Full article

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22 pages, 6094 KB

Open AccessArticle

Susceptibility to Secondary Bacterial Infections in Growing Rabbits Exposed to Ochratoxin A and Protected or Not by Herbal Supplements

by Kalina Zhivkova, Stoycho Stoev, Vladimir Petrov and Vesselin Ivanov

Toxins 2025, 17(10), 507; https://doi.org/10.3390/toxins17100507 - 15 Oct 2025

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Abstract

The protective effects of the herbal feed supplements Silybum marianum, Silymarin, Withania somnifera, and Centella asiatica against ochratoxin A (OTA) toxicity were studied in 48 New Zealand White rabbits (37-day-old) during an 80-day experiment. OTA was given at 2 ppm, whereas Silybum [...] Read more.

The protective effects of the herbal feed supplements Silybum marianum, Silymarin, Withania somnifera, and Centella asiatica against ochratoxin A (OTA) toxicity were studied in 48 New Zealand White rabbits (37-day-old) during an 80-day experiment. OTA was given at 2 ppm, whereas Silybum marianum, Silymarin, Withania somnifera, and Centella asiatica were given at feed levels of 5000 ppm, 25,000 ppm, 4000 ppm, and 4600 ppm, respectively. All rabbits were immunized against Rabbit Hemorrhagic Disease Virus (RHDV). OTA was found to induce an immunosuppressive effect on the humoral immune response. Reliable protection against OTA-provoked immunosuppression by Silimarin and Withania somnifera was found. The OTA-induced immunosuppression was responsible for secondary bacterial infection (pasteurellosis) and the death of two rabbits from the OTA-exposed group and one rabbit each from the groups protected with Silybum marianum and Centella asiatica. A decreased body weight was found in rabbits exposed to OTA, but the decrease was slighter in the rabbits protected with herbal supplements. The target organs damaged by OTA exposure were the liver, kidneys, and spleen, while weaker lesions were found in other internal organs, except in the cases of secondary pasteurellosis, in which the strongest damage was found in the lung. All investigated herbal supplements appeared to have stronger protective effects against OTA-induced damage to the kidneys and liver, with slightly protective effects observed in the lungs, myocardium, spleen, brain, intestine, testicles, and ovaries. Full article

(This article belongs to the Section Mycotoxins)

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21 pages, 8380 KB

Open AccessArticle

Comprehensive Study of Some Cyanobacteria in Moscow Waterbodies (Russia), Including Characteristics of the Toxigenic Microcystis aeruginosa Strains

by Elena Kezlya, Elina Mironova, Ekaterina Chernova, Maria Gololobova, Andrei Mironov, Ekaterina Voyakina, Yevhen Maltsev, Dina Snarskaya and Maxim Kulikovskiy

Toxins 2025, 17(10), 506; https://doi.org/10.3390/toxins17100506 - 14 Oct 2025

Viewed by 358

Abstract

The current study focuses on the diversity, distribution and toxic potential of cyanobacteria in the waterbodies of Moscow, Russia. The research involves the sampling of natural and artificial water environments situated within the Moscow city agglomeration, including the waterbodies of recreational importance. A [...] Read more.

The current study focuses on the diversity, distribution and toxic potential of cyanobacteria in the waterbodies of Moscow, Russia. The research involves the sampling of natural and artificial water environments situated within the Moscow city agglomeration, including the waterbodies of recreational importance. A total of 20 strains of cyanobacteria, namely representatives of Anabaena, Aphanizomenon, Argonema, Dolichospermum, Microcystis and Woronichinia, are isolated from the collected samples. The morphology of the newly obtained strains is analyzed through light microscopy. The results of morphological identification are compared to the molecular data. The molecular phylogeny of the cyanobacterial strains is assessed on the basis of 16S rRNA sequencing. The detection of cyanotoxin-producing genes through PCR reveals two strains of Microcystis aeruginosa capable of microcystin synthesis. Further analysis using HPLC-HRMS demonstrates that microcystin production includes a high proportion (20–28%) of exceptionally toxic microcystin–leucine arginine compounds. Hereby, we discuss the morphology and phylogeny of the analyzed strains and provide comments on the toxic potential of cyanobacteria within the waterbodies of Moscow. Full article

(This article belongs to the Special Issue Prospective Studies on Harmful Cyanobacteria and Cyanotoxins)

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2 pages, 628 KB

Open AccessCorrection

Correction: Thenuwara et al. Recent Advancements in Lateral Flow Assays for Food Mycotoxin Detection: A Review of Nanoparticle-Based Methods and Innovations. Toxins 2025, 17, 348

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

Toxins 2025, 17(10), 505; https://doi.org/10.3390/toxins17100505 - 14 Oct 2025

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Abstract

Figure/Legend [...] Full article

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

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15 pages, 1416 KB

Open AccessArticle

Profiling the Paralytic Effects and Lethality of Cone Snail Venom Toxins Using Nanofractionation Analytics with In Vivo Zebrafish Larvae Assays

by Jeroen Kool, Arif Arrahman, Haifeng Xu, Jiaxing Liu, Richard J. Lewis, Christian Tudorache and Fernanda C. Cardoso

Toxins 2025, 17(10), 504; https://doi.org/10.3390/toxins17100504 - 13 Oct 2025

Viewed by 488

Abstract

This study presents nanofractionation analytics coupled with in vivo profiling of zebrafish embryo paralysis and lethality in response to toxins in cone snail venoms. The focus of this study is on the development of this approach using venoms of Conus marmoreus, Conus [...] Read more.

This study presents nanofractionation analytics coupled with in vivo profiling of zebrafish embryo paralysis and lethality in response to toxins in cone snail venoms. The focus of this study is on the development of this approach using venoms of Conus marmoreus, Conus ebraeus, and Conus bandanus. In brief, cone snail venoms were separated using reversed-phase chromatography following high-resolution nanofractionation on microplates with parallel mass spectrometry, enabled via a post-column flow split. All collected fractions were dried overnight, followed by assays on zebrafish embryos. For the paralysis assessment, we monitored swimming behavior and swimming distance and found that exposure to cone snail toxins led to paralysis and decreased movement and swim distance. To correlate the masses of eluted toxins with their paralyzing effects and potency, we compared the fractionation retention time versus normalized swimming distance. This allowed identification of the masses of toxins with paralyzing bioactivity, which were predominantly conopeptides. To assess lethality, zebrafish embryos were exposed to fractionated toxins for 24 h, after which they were inspected. The lethal doses and correlated toxins were identified by comparing retention times of fractionation versus the lethal dose values calculated for each fraction. We found that the most lethal venom was from C. bandanus, displaying the largest number of lethal peptides, followed by C. marmoreus and C. ebraeus. On the other hand, the most paralytic venom was from C. ebraeus, presenting a higher number of peptides with non-lethal paralytic effects, followed by C. bandanus and C. marmoreus. This study provides a pipeline to rapidly identify paralytic and lethal cone snail venom toxins using the zebrafish embryo model. Full article

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

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11 pages, 266 KB

Open AccessReview

Botulinum Toxin Treatment in Hereditary Spastic Paraplegia—A Comprehensive Review and Update

by Bahman Jabbari, Samira Comtesse and Fattaneh Tavassoli

Toxins 2025, 17(10), 503; https://doi.org/10.3390/toxins17100503 - 13 Oct 2025

Viewed by 469

Abstract

Using Medline and Scopus as search engines, we identified reports of 10 clinical studies (published up to 1 September 2025) on botulinum neurotoxin therapy for hereditary spastic paraplegia (HSP). Nine studies were conducted in adults and one in children. Only one of the [...] Read more.

Using Medline and Scopus as search engines, we identified reports of 10 clinical studies (published up to 1 September 2025) on botulinum neurotoxin therapy for hereditary spastic paraplegia (HSP). Nine studies were conducted in adults and one in children. Only one of the ten studies was double-blind and placebo-controlled. The search strategy included only articles published in English and articles providing basic information such as the type of the study, type and dose of the toxin and results of the treatment. Articles not in English, case reports and review articles were excluded. A total of 258 patients were included across all studies. The injected toxin in the open-label studies was botulinumtoxin-A (Botox or Dysport or Xeomin), whereas in the blinded study, the investigators used Prosigne. All open-label studies, which used FDA approved botulinumtoxin-A neurotoxins, demonstrated a degree of motor and non-motor improvement, whereas treatment with Prosigne did not improve patients’ function. The possible reasons for this discrepancy between the blinded study and the open-label studies are discussed. We found no studies on the effect of BoNTs on bladder dysfunction in HSP. There is a need for double-blind, placebo-controlled studies assessing the efficacy of FDA-approved botulinum neurotoxins in children and adults affected by hereditary spastic paraparesis. Such studies should also investigate the effect(s) of early botulinum neurotoxin therapy in this disorder. The novelty of this review is that it represents a comprehensive and critical literature review on this subject, with no other studies of this kind published previously. It also includes data not present in previous reviews of this subject. Full article

(This article belongs to the Special Issue Botulinum Toxin in Neuro-Rehabilitation: Expanding Horizons in Spasticity and Beyond)

23 pages, 10020 KB

Open AccessArticle

Microbiological and Mycotoxicological Quality of Stored Wheat, Wholemeal Flour and Bread: The Impact of Extreme Weather Events in Romania in the 2024 Summer

by Valeria Gagiu, Elena Mirela Cucu (Chirtu), Elena Iulia Lazar (Banuta), Cristian Mihai Pomohaci, Alina Alexandra Dobre, Gina Pusa Pirvu, Oana Alexandra Oprea, Cristian Lazar, Elena Mateescu and Nastasia Belc

Toxins 2025, 17(10), 502; https://doi.org/10.3390/toxins17100502 - 11 Oct 2025

Viewed by 963

Abstract

This study examines the effects of the extreme drought and heatwaves that occurred in Romania during the summer of 2024 on the microbiological and mycotoxicological quality of wheat (Triticum aestivum) stored until April 2025, as well as on the quality of [...] Read more.

This study examines the effects of the extreme drought and heatwaves that occurred in Romania during the summer of 2024 on the microbiological and mycotoxicological quality of wheat (Triticum aestivum) stored until April 2025, as well as on the quality of wholemeal flour and bread derived from it. Comparative analyses were conducted against the contamination in wheat harvested in 2024. The hot and dry conditions significantly influenced the microbial and mycotoxicological contamination of both freshly harvested and stored wheat, as well as the derived flour and bread, due to their notably reduced moisture content and water activity. Although levels of total fungi, Fusarium-damaged kernels, and mycotoxins deoxynivalenol, aflatoxin B1, and ochratoxin A remained well below regulatory thresholds, higher contamination was observed in Transylvania and Moldavia—particularly in the Curvature Carpathians, likely due to their cooler and wetter microclimates. The observed quality changes were strongly associated with alterations in physico-chemical, rheological, and colorimetric parameters, posing potential economic challenges for the milling and baking industries. The study recommends implementing integrated regional strategies to enhance wheat resilience, optimize production systems, and improve contamination control in response to increasing climate stress across Southeastern Europe. Full article

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

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16 pages, 2289 KB

Open AccessFeature PaperArticle

RelabotulinumtoxinA, a Ready-to-Use Formulation Neuromodulator Manufactured with PEARL™ Technology to Maintain High Biological and Specific Activity

by Ulf Ståhl, Emilia Lekholm, Emil Hamnevik, Robert Fredriksson, Sachin M. Shridharani, Keywan Taghetchian, Joel L. Cohen, Mark S. Nestor and Åsa Liljegren Sundberg

Toxins 2025, 17(10), 501; https://doi.org/10.3390/toxins17100501 - 9 Oct 2025

Viewed by 584

Abstract

Most botulinum toxin A (BoNT-A) products for esthetic use require reconstitution before administration. Ready-to-use relabotulinumtoxinA is a liquid manufactured using Precipitation-free Extraction and Activity-preserving, Refined Liquid (PEARL™) Technology from a proprietary C. botulinum type A1 strain. We examined the in vitro characteristics of [...] Read more.

Most botulinum toxin A (BoNT-A) products for esthetic use require reconstitution before administration. Ready-to-use relabotulinumtoxinA is a liquid manufactured using Precipitation-free Extraction and Activity-preserving, Refined Liquid (PEARL™) Technology from a proprietary C. botulinum type A1 strain. We examined the in vitro characteristics of relabotulinumtoxinA. The specific BoNT-A1 potency remained consistent throughout drug substance manufacturing (1.9 × 10⁸–2.2 × 10⁸ LD₅₀ mouse potency units/mg of BoNT-A1, four fractions sampled). Using glabellar line (GL) on-label doses, relabotulinumtoxinA liquid product was compared with powder onabotulinumtoxinA using the following: BoNT-A1 amount based on ELISA; specific enzyme activity based on SNAP-25 cleavage by a fluorescence resonance energy transfer-based assay (BoTest^®); biological activity (binding, internalization, and SNAP-25 cleavage over time) using a cell-based assay. RelabotulinumtoxinA contained more BoNT-A1 per on-label GL dose (0.27 ng) than onabotulinumtoxinA (0.18 ng), had higher enzyme activity (53 vs. 29 BoTest^® units) per GL dose, and had higher specific activity per pg BoNT-A, with onabotulinumtoxinA displaying 81% of the specific activity of relabotulinumtoxinA. In vitro, relabotulinumtoxinA demonstrated higher biological activity and earlier onset of SNAP-25-cleavage than onabotulinumtoxinA. PEARL^TM Technology thus produces high-quality BoNT-A1 with high specific enzyme and biological activities, which may explain the clinical performance of relabotulinumtoxinA in Phase 3 clinical trials examining treatment of GLs and/or LCLs. Full article

(This article belongs to the Section Bacterial Toxins)

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20 pages, 3594 KB

Open AccessArticle

Ganglioside Binding Assay: A Complementary Approach for Enhanced Tetanus Toxoid Quality Control

by Yuki Tanoue, Chie Shitada, Mariko Nakamichi, Naomi Nakamichi, Chiyomi Sakamoto, Hyun Kim, Kohsuke Kumeda, Masaki Ochiai, Susumu Yamaori, Mitsutoshi Senoh and Motohide Takahashi

Toxins 2025, 17(10), 500; https://doi.org/10.3390/toxins17100500 - 9 Oct 2025

Viewed by 392

Abstract

Vaccine quality control has long relied on animal testing, which involves time, cost, and ethical constraints. This study introduces a ganglioside binding (GB) assay as a complementary in vitro screening tool for tetanus toxoid quality control, which was validated in a single-laboratory environment [...] Read more.

Vaccine quality control has long relied on animal testing, which involves time, cost, and ethical constraints. This study introduces a ganglioside binding (GB) assay as a complementary in vitro screening tool for tetanus toxoid quality control, which was validated in a single-laboratory environment as a foundational proof-of-concept. The assay reproduces tetanus toxin binding to gangliosides on microplates using simplified procedures. Validation with samples at different inactivation stages showed excellent linearity (0.0002–0.0156 Lf/mL), reproducibility, and a strong correlation with Ramon’s flocculation (R² = 0.999). The assay clearly distinguished between toxins and toxoids, with the toxoid results remaining at control levels. The time-course inactivation samples were consistent with the animal tests: partially inactivated samples (days 1–3) showed significant GB activity (p < 0.001) and caused 100% mortality, whereas samples from day 4 onward showed no activity and zero mortality. These findings demonstrate that the GB assay reliably differentiates active toxins from toxoids, which aligns with in vivo outcomes. The practical advantages include a simplified protocol, reduced complexity, and improved efficiency for routine testing of samples. As a complementary screening approach, this single-laboratory validation supports the 3Rs principle by demonstrating the potential for reducing animal use while ensuring quality assurance. Broader applicability requires multicenter validation and cross-reactivity, and multicenter validation is ongoing. Full article

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16 pages, 1361 KB

Open AccessArticle

Renal Effects of Sulfated Polysaccharides from the Seaweed Gracilaria cornea

by Terentia Batista Sá Norões, Sophia Moinhos, Helena Serra Azul Monteiro, Alice Maria Costa Martins, Ricardo Parente Garcia Vieira and Claudio Gleidiston Silva

Toxins 2025, 17(10), 499; https://doi.org/10.3390/toxins17100499 - 9 Oct 2025

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Abstract

Sulfated polysaccharides (SPs) are abundant in seaweed and have several industrial and biomedical applications, but their renal effects remain unclear. This study evaluated the effects of total sulfated polysaccharides (TSPs) from Gracilaria cornea using an isolated rat kidney perfusion model. TSP at 3 [...] Read more.

Sulfated polysaccharides (SPs) are abundant in seaweed and have several industrial and biomedical applications, but their renal effects remain unclear. This study evaluated the effects of total sulfated polysaccharides (TSPs) from Gracilaria cornea using an isolated rat kidney perfusion model. TSP at 3 µg/mL increased perfusion pressure and renal vascular resistance at 90–120 min, while 4.5 µg/mL induced earlier and more pronounced changes (from 60 min). Urinary flow decreased at 1 µg/mL (90 min) but increased at 4.5 µg/mL (90–120 min). Sodium transport was reduced at all concentrations, whereas potassium and chloride transport remained unchanged. Histological analysis revealed protein deposits in tubules and urinary space, indicating tubular injury. In vitro, TSP reduced MDCK cell viability in a concentration-dependent manner and induced apoptosis, with some cells progressing to secondary necrosis. In conclusion, TSP altered renal physiology and morphology and triggered apoptotic pathways in renal cells, highlighting the need for further mechanistic and translational studies. Full article

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18 pages, 966 KB

Open AccessArticle

Computational Modelling Suggests Bacteriostatic Saline Does Not Reverse Botulinum Toxin-Induced Brow Ptosis

by Eqram Rahman, Alain Michon, Parinitha Rao, A. Q. M. Omar Sharif, William Richard Webb and Jean D. A. Carruthers

Toxins 2025, 17(10), 498; https://doi.org/10.3390/toxins17100498 - 7 Oct 2025

Viewed by 771

Abstract

Anecdotal reports have recently circulated suggesting that intramuscular injection of bacteriostatic saline (BS)—which contains benzyl alcohol (BnOH)—can reverse botulinum toxin type A (BoNTA)-induced brow ptosis. Given the well-established intracellular persistence of BoNTA’s light chain and its irreversible cleavage of SNAP-25, such rapid functional [...] Read more.

Anecdotal reports have recently circulated suggesting that intramuscular injection of bacteriostatic saline (BS)—which contains benzyl alcohol (BnOH)—can reverse botulinum toxin type A (BoNTA)-induced brow ptosis. Given the well-established intracellular persistence of BoNTA’s light chain and its irreversible cleavage of SNAP-25, such rapid functional recovery challenges existing pharmacological understanding. This study employed high-resolution pharmacokinetic/pharmacodynamic (PK/PD) modelling using the AesthetiSim™ platform to systematically evaluate this hypothesis. A total of 30,000 virtual patients were randomized to receive BoNTA alone, BoNTA followed by BS injection, or BoNTA followed by normal saline (NS) at Day 7. The model incorporated BoNTA diffusion, internalization, SNAP-25 cleavage, neuromuscular output, and transient BS effects on membrane permeability and endosomal trafficking. Simulated recovery trajectories were tracked over 90 days. The primary outcome, time to 80% restoration of baseline frontalis muscle force (T₈₀), averaged 42.0 days in the BoNTA-only group and 35.5 days in the BS group (Δ = −6.5 days; p < 0.001). Only 13.9% of BS-treated patients reached the T₈₀ threshold by Day 30. Partial reactivation (T₃₀) occurred earlier with BS (21.8 ± 5.3 days vs. 27.3 ± 4.9 days), and the area under the effect curve (AUEC) was increased by 9.7%, reflecting higher overall muscle function over time. In molecular simulations, BnOH produced a minor rightward shift in the BoNTA–SNAP-25 dissociation curve, but receptor occupancy remained above 90% at therapeutic toxin concentrations, suggesting no meaningful impairment of binding affinity. A global Sobol sensitivity analysis demonstrated that the primary driver of recovery kinetics was intracellular LC degradation (49% of T₈₀ variance), while BS-modulated extracellular parameters collectively contributed less than 20%. These findings indicate that BS does not reverse the molecular action of BoNTA but may transiently influence recovery kinetics via non-receptor-mediated pathways such as increased membrane permeability or altered vesicular trafficking. The magnitude and variability of this effect do not support the notion of a true pharmacologic reversal. Instead, these results emphasize the need for mechanistic scrutiny when evaluating rapid-reversal claims, particularly those propagated through anecdotal or social media channels without supporting biological plausibility. Full article

(This article belongs to the Section Bacterial Toxins)

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57 pages, 4553 KB

Open AccessReview

Properties and Pharmacology of Scorpion Toxins and Their Biotechnological Potential in Agriculture and Medicine

by Cháriston André Dal Belo, Stephen Hyslop and Célia Regina Carlini

Toxins 2025, 17(10), 497; https://doi.org/10.3390/toxins17100497 - 7 Oct 2025

Viewed by 931

Abstract

Scorpion venoms contain a wide range of toxins that interact with a variety of target molecules (ion channels, receptors and enzymes) associated with synaptic transmission, action potential propagation, cardiac function, hemostasis and other physiological systems. Scorpion toxins are also active towards bacteria, viruses, [...] Read more.

Scorpion venoms contain a wide range of toxins that interact with a variety of target molecules (ion channels, receptors and enzymes) associated with synaptic transmission, action potential propagation, cardiac function, hemostasis and other physiological systems. Scorpion toxins are also active towards bacteria, viruses, fungi and parasites. Such interactions make scorpion toxins useful lead molecules for developing compounds with biotechnological and therapeutic applications, and as tools for cell biology. In addition, scorpion toxins act as insectotoxins, with promising applications as insecticides. This review describes the range of scorpion toxins and discusses their usefulness for the development of insecticides and therapeutic drugs. Full article

(This article belongs to the Special Issue New Insights into Structure and Function of Arthropod Toxins and Other Toxins: Tribute to Professor Hervé Rochat)

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20 pages, 6584 KB

Open AccessArticle

Compromised Regeneration, Damage to Blood Vessels and the Endomysium Underpin Permanent Muscle Damage Induced by Puff Adder (Bitis arietans) Venom

by Sodiq Opeyemi Adeyemi, Nicholas John Richards, Ali Alqallaf, Husain Bin Haidar, Mustafa Jawad Jalil Al-Asady, Jarred Williams, José R. Almeida and Ketan Patel

Toxins 2025, 17(10), 496; https://doi.org/10.3390/toxins17100496 - 6 Oct 2025

Viewed by 637

Abstract

The puff adder (Bitis arietans) is a clinically relevant viper species found throughout Africa, and it is responsible for a greater incidence of health-related envenomations than all other snake species on the continent combined. Unresolved skeletal muscle damage is a common [...] Read more.

The puff adder (Bitis arietans) is a clinically relevant viper species found throughout Africa, and it is responsible for a greater incidence of health-related envenomations than all other snake species on the continent combined. Unresolved skeletal muscle damage is a common consequence of B. arietans envenomation that can result in long-term morbidity and even death. Antivenom treatment can mitigate the systemic effects of the venom but offers little protection against local tissue damage. Identifying the mechanisms through which B. arietans venom induces tissue damage and impedes skeletal muscle regeneration could identify possible treatment alternatives that could help alleviate the long-term consequences of envenomation. Skeletal muscle has an innate ability to regenerate, but constituents within the venom can impede multiple stages of this regeneration process. In this study, we employed a combination of biochemical analyses, cell-based assays, and in vivo experiments to assess the toxicological implications of B. arietans envenomation and its impacts on key processes of regeneration. Our findings demonstrate that the pathological characteristics of permanent muscle damage resulting from B. arietans envenomation may be attributed to the venom’s effects on muscle stem cell precursors, the extracellular matrix (ECM), and the influence of blood-borne proteins that promote fibrosis. Full article

(This article belongs to the Special Issue Snake Bite and Related Injury)

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29 pages, 2554 KB

Open AccessReview

Toxic Alexandrium Treatment in Western Australia: Investigating the Efficacy of Modified Nano Clay

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

Toxins 2025, 17(10), 495; https://doi.org/10.3390/toxins17100495 - 4 Oct 2025

Viewed by 573

Abstract

Alexandrium spp. blooms produce a range of toxins, including spirolides, goniodomins, and paralytic shellfish toxins (PSTs). Of these, PSTs are the most impactful due to their high affinity for voltage-gated sodium ion channels in nerve cell membranes. This interaction can cause neurological effects [...] Read more.

Alexandrium spp. blooms produce a range of toxins, including spirolides, goniodomins, and paralytic shellfish toxins (PSTs). Of these, PSTs are the most impactful due to their high affinity for voltage-gated sodium ion channels in nerve cell membranes. This interaction can cause neurological effects such as paralysis and, in severe cases, may lead to death. Given the implications of Alexandrium blooms on public health, all mitigation, prevention, and treatment strategies aim to reduce their socioeconomic impacts. However, monitoring harmful algal blooms remains difficult due to confounding influences such as pollution, climate change, and the inherent variability of environmental conditions. These factors can complicate early detection and management efforts, especially as the intensity and frequency of blooms continue to rise, further exacerbating their socioeconomic consequences. This review offers insights into several management approaches to prevent and control Alexandrium blooms, focusing on modified nano-clays as a promising emergency mitigation measure for low-density toxic algal blooms, especially in areas predominantly used for recreational fishing. However, it is recommended that treatment be coupled with monitoring to alleviate reliance on treatment alone. Full article

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

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19 pages, 1292 KB

Open AccessReview

Ricin and Abrin in Biosecurity: Detection Technologies and Strategic Responses

by Wojciech Zajaczkowski, Ewelina Bojarska, Elwira Furtak, Michal Bijak, Rafal Szelenberger, Marcin Niemcewicz, Marcin Podogrocki, Maksymilian Stela and Natalia Cichon

Toxins 2025, 17(10), 494; https://doi.org/10.3390/toxins17100494 - 3 Oct 2025

Viewed by 724

Abstract

Plant-derived toxins such as ricin and abrin represent some of the most potent biological agents known, posing significant threats to public health and security due to their high toxicity, relative ease of extraction, and widespread availability. These ribosome-inactivating proteins (RIPs) have been implicated [...] Read more.

Plant-derived toxins such as ricin and abrin represent some of the most potent biological agents known, posing significant threats to public health and security due to their high toxicity, relative ease of extraction, and widespread availability. These ribosome-inactivating proteins (RIPs) have been implicated in politically and criminally motivated events, underscoring their critical importance in the context of biodefense. Public safety agencies, including law enforcement, customs, and emergency response units, require rapid, sensitive, and portable detection methods to effectively counteract these threats. However, many existing screening technologies lack the capability to detect biotoxins unless specifically designed for this purpose, revealing a critical gap in current biodefense preparedness. Consequently, there is an urgent need for robust, field-deployable detection platforms that operate reliably under real-world conditions. End-users in the security and public health sectors demand analytical tools that combine high specificity and sensitivity with operational ease and adaptability. This review provides a comprehensive overview of the biochemical characteristics of ricin and abrin, their documented misuse, and the challenges associated with their detection. Furthermore, it critically assesses key detection platforms—including immunoassays, mass spectrometry, biosensors, and lateral flow assays—focusing on their applicability in operational environments. Advancing detection capabilities within frontline services is imperative for effective prevention, timely intervention, and the strengthening of biosecurity measures. Full article

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15 pages, 1129 KB

Open AccessArticle

Building Sub-Saharan African PBPK Populations Reveals Critical Data Gaps: A Case Study on Aflatoxin B1

by Orphélie Lootens, Marthe De Boevre, Sarah De Saeger, Jan Van Bocxlaer and An Vermeulen

Toxins 2025, 17(10), 493; https://doi.org/10.3390/toxins17100493 - 3 Oct 2025

Viewed by 868

Abstract

Physiologically based pharmacokinetic (PBPK) models allow to simulate the behaviour of compounds in diverse physiological populations. However, the categorization of individuals into distinct populations raises questions regarding the classification criteria. In previous research, simulations of the pharmacokinetics of the mycotoxin aflatoxin B1 (AFB1), [...] Read more.

Physiologically based pharmacokinetic (PBPK) models allow to simulate the behaviour of compounds in diverse physiological populations. However, the categorization of individuals into distinct populations raises questions regarding the classification criteria. In previous research, simulations of the pharmacokinetics of the mycotoxin aflatoxin B1 (AFB1), were performed in the black South African population, using PBPK modeling. This study investigates the prevalence of clinical CYP450 phenotypes (CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4/5) across Sub-Saharan Africa (SSA), to determine the feasibility of defining SSA as a single population. SSA was subdivided into Central, East, South and West Africa. The phenotype data were assigned to the different regions and a fifth SSA group was composed of all regions’ weighted means. Available data from literature only covered 7.30% of Central, 56.9% of East, 38.9% of South and 62.9% of West Africa, clearly indicating critical data gaps. A pairwise proportion test was performed between the regions on enzyme phenotype data. When achieving statistical significance (p < 0.05), a Cohen’s d-test was performed to determine the degree of the difference. Next, per region populations were built using SimCYP starting from the available SSA based SouthAfrican_Population FW_Custom population, supplemented with the phenotype data from literature. Simulations were performed using CYP probe substrates in all populations, and derived PK parameters (C_max, T_max, AUC_ss and CL) were plotted in bar charts. Significant differences between the African regions regarding CYP450 phenotype frequencies were shown for CYP2B6, CYP2C19 and CYP2D6. Limited regional data challenge the representation of SSA populations in these models. The scarce availability of in vivo data for SSA regions restricted the ability to fully validate the developed PBPK populations. However, observed literature data from specific SSA regions provided partial validation, indicating that SSA populations should ideally be modelled at a regional level rather than as a single entity. The findings, emerging from the initial AFB1-focused PBPK work, underscore the need for more extensive and region-specific data to enhance model accuracy and predictive value across SSA. Full article

(This article belongs to the Special Issue Mycotoxins in Food and Feeds: Human Health and Animal Nutrition)

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18 pages, 723 KB

Open AccessReview

Mycotoxin Research in Algeria: A Comprehensive Review of Trends, Challenges, and Future Directions

by Yamina Ben Miri, Imene Chentir, Aldjia Taoudiat, Amina Benabdallah and Marta Herrera

Toxins 2025, 17(10), 492; https://doi.org/10.3390/toxins17100492 - 3 Oct 2025

Viewed by 473

Abstract

This review offers a comprehensive overview of mycotoxin research and related publications in Algeria, outlining key trends, existing challenges, and prospects for future advancement. Despite limited exploration of mycotoxins in Algeria, researchers have made significant progress in understanding mycotoxin contamination and its effects [...] Read more.

This review offers a comprehensive overview of mycotoxin research and related publications in Algeria, outlining key trends, existing challenges, and prospects for future advancement. Despite limited exploration of mycotoxins in Algeria, researchers have made significant progress in understanding mycotoxin contamination and its effects on food safety and public health. The review delves into both research achievements and the challenges encountered in resource-limited settings, while also exploring strategies employed to surmount these obstacles. Through an analysis of existing literature, key themes emerge regarding mycotoxin identification, detection methods, mitigation strategies, and their implications. The importance of collaborative efforts among academia and government agencies is underscored as pivotal in addressing mycotoxin-related challenges. Moreover, this review identified gaps in current research and offered recommendations for future investigations, aiming to advance mycotoxin research in Algeria and beyond. Full article

(This article belongs to the Special Issue Mycotoxins in Food Chain: Occurrence, Analysis and Risk Management)

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17 pages, 2248 KB

Open AccessArticle

Expression of L-Amino Acid Oxidase (Ml-LAAO) from the Venom of the Micrurus lemniscatus Snake in a Mammalian Cell System

by Ari Junio de Oliveira Costa, Alessandra Matavel, Patricia Cota Campos, Jaqueline Leal dos Santos, Ana Caroline Zampiroli Ataide, Sophie Yvette Leclercq, Valéria Gonçalves de Alvarenga, Sergio Caldas, William Castro-Borges and Márcia Helena Borges

Toxins 2025, 17(10), 491; https://doi.org/10.3390/toxins17100491 - 2 Oct 2025

Viewed by 456

Abstract

Animal venoms are rich in bioactive molecules with promising biotechnological potential. They comprise both protein and non-protein toxins. Among the protein toxins are enzymes, such as phospholipases A₂, proteases and L-amino acid oxidases (LAAOs). LAAOs exhibit antimicrobial, antiparasitic, antiviral, and anticancer [...] Read more.

Animal venoms are rich in bioactive molecules with promising biotechnological potential. They comprise both protein and non-protein toxins. Among the protein toxins are enzymes, such as phospholipases A₂, proteases and L-amino acid oxidases (LAAOs). LAAOs exhibit antimicrobial, antiparasitic, antiviral, and anticancer effects, making them potential candidates for biotechnological applications. These activities are linked to their ability to catalyze oxidative reactions that convert L-amino acids into α-keto acids, releasing ammonia and hydrogen peroxide, which contribute to the immune response, pathogen elimination, and oxidative stress. However, in snakes of the Micrurus genus, LAAOs generally represent a small portion of the venom (up to ~7%), which limits their isolation and study. To overcome this, the present study aimed to produce Ml-LAAO, the enzyme from Micrurus lemniscatus, through heterologous expression in mammalian cells. The gene sequence was inferred from its primary structure and synthesized into the pSecTag2B vector for expression in HEK293T cells. After purification using a His Trap-HP column, the presence of recombinant Ml-LAAO (Ml-LAAOrec) was confirmed by Western blot and mass spectrometry, validating its identity. These results support successful recombinant expression of Ml-LAAO and highlight its potential for scalable production and future biotechnological applications. Full article

(This article belongs to the Special Issue Biochemistry, Pathology and Applications of Venoms)

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16 pages, 2336 KB

Open AccessArticle

Bioremediation of Contaminated Water: The Potential of Aquatic Plants Ceratophyllum demersum and Pistia stratiotes Against Toxic Bloom

by Fatma Zohra Tamer, Hadjer Zaidi, Hichem Nasri, Larisa Lvova, Nada Nouri, Fateh Sedrati, Amina Amrani, Nassima Beldjoudi and Xi Li

Toxins 2025, 17(10), 490; https://doi.org/10.3390/toxins17100490 - 2 Oct 2025

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Abstract

Toxic cyanobacteria, including Microcystis, produce harmful toxins that affect aquatic ecosystems and human health. Biotreatment using macrophytes shows promise in mitigating these blooms. This study investigates the bioaccumulation dynamics and biochemical responses of two aquatic macrophytes, Pistia stratiotes and Ceratophyllum demersum, [...] Read more.

Toxic cyanobacteria, including Microcystis, produce harmful toxins that affect aquatic ecosystems and human health. Biotreatment using macrophytes shows promise in mitigating these blooms. This study investigates the bioaccumulation dynamics and biochemical responses of two aquatic macrophytes, Pistia stratiotes and Ceratophyllum demersum, in removing microcystin from contaminated water. P. stratiotes showed high initial bioaccumulation rates with rapid microcystin uptake, which is effective for short-term bioremediation. C. demersum has shown stable bioaccumulation. Biochemical analyses have revealed the activation of plant antioxidant defenses, with both macrophytes showing an increase in carotenoids, glutathione (GSH), and antioxidant enzymes such as superoxide dismutase (SOD) and glutathione-S-transferase (GST) concentrations. In particular, C. demersum has maintained higher antioxidant levels, contributing to its sustained capacity and resilience. Fluctuations in malondialdehyde (MDA) indicated oxidative stress, with P. stratiotes managing such stress through its defenses. Principal Component Analysis (PCA) supports these findings: Pistia’s first two components explained 25.09% and 20.71% of the variance, with Carotenoid and Chl contributing strongly to PC1, and MDA and GST influencing both components. For C. demersum, PC1 and PC2 explained 21.79% and 19.78% of the variance, with Carotenoid and Chl a being major contributors, while SOD and GSH played significant roles in sample differentiation. Integrating both plants into bioremediation strategies could optimize microcystin removal: P. stratiotes offers rapid initial detoxification, while C. demersum ensures continuous, long-term remediation. This combined approach enhances the efficiency and sustainability of phytoremediation. Future research should optimize environmental conditions and explore synergistic effects among multiple plant species for more effective and sustainable bioremediation solutions. Full article

(This article belongs to the Section Bacterial Toxins)

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14 pages, 1349 KB

Open AccessArticle

ProToxin, a Predictor of Protein Toxicity

by Yang Yang, Haohan Zhang and Mauno Vihinen

Toxins 2025, 17(10), 489; https://doi.org/10.3390/toxins17100489 - 1 Oct 2025

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Abstract

Toxins are naturally poisonous small compounds, peptides and proteins that are produced in all three kingdoms of life. Venoms are animal toxins and can contain even hundreds of different compounds. Numerous approaches have been used to detect toxins, including prediction methods. We developed [...] Read more.

Toxins are naturally poisonous small compounds, peptides and proteins that are produced in all three kingdoms of life. Venoms are animal toxins and can contain even hundreds of different compounds. Numerous approaches have been used to detect toxins, including prediction methods. We developed a novel machine learning-based predictor for detecting protein toxins from their sequences. The gradient boosting method was trained on carefully selected training data. Initially, we tested 2614 features, which were reduced to 88 after a comprehensive feature selection procedure. Out of the four tested algorithms, XGBoost was chosen to train the final predictor. Comparison to available predictors indicated that ProToxin showed significant improvement compared to state-of-the-art predictors. On a blind test dataset, the accuracy was 0.906, the Matthews correlation coefficient was 0.796, and the overall performance measure was 0.796. ProToxin is a fast and efficient method and is freely available. It can be used for small and large numbers of sequences. Full article

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21 pages, 4001 KB

Open AccessArticle

Exploring the Venom Diversity of Australian Taipans: Comparative Characterization of Oxyuranus microlepidotus and Oxyuranus scutellatus

by Guilherme Gonelli Paz, Patrick Jack Spencer, Daniel Carvalho Pimenta and Emidio Beraldo-Neto

Toxins 2025, 17(10), 488; https://doi.org/10.3390/toxins17100488 - 1 Oct 2025

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Abstract

The genus Oxyuranus, which includes some of the most venomous snakes in the world, presents a complex venom composition with potent neurotoxic and procoagulant effects. This study provides a comparative proteomic analysis of the venom of Oxyuranus microlepidotus (Inland Taipan) and Oxyuranus [...] Read more.

The genus Oxyuranus, which includes some of the most venomous snakes in the world, presents a complex venom composition with potent neurotoxic and procoagulant effects. This study provides a comparative proteomic analysis of the venom of Oxyuranus microlepidotus (Inland Taipan) and Oxyuranus scutellatus (Coastal Taipan), aiming to elucidate the molecular basis underlying their distinct toxicological profiles. Using high-resolution chromatographic fractionation and LC-MS/MS, we identified a core set of nine protein families shared between both species, including phospholipases A₂ (PLA₂), three-finger toxins (3FTx), natriuretic peptides (NTP), nerve growth factors (NGF), and prothrombin activators (PTA). O. microlepidotus venom exhibited greater diversity of 3FTxs and unique protein families, such as Waprin and 5′-nucleotidases, suggesting lineage-specific functional adaptations. Quantitative analysis revealed a greater relative abundance of PLA₂s in O. scutellatus (66%) compared to O. microlepidotus (47%), whereas 3FTXs were more prominent in O. microlepidotus (33% vs. 9%). These interspecific differences likely underlie the distinct clinical manifestations of envenomation and reflect evolutionary divergence in the venom composition. Our findings provide molecular insights into taipan venom complexity and highlight novel toxin candidates with potential biomedical applications in neurobiology, hemostasis, and anti-infective therapy. Full article

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

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18 pages, 1671 KB

Open AccessArticle

Toxigenic Aspergillus Diversity and Mycotoxins in Organic Spanish Grape Berries

by Clara Melguizo, Andrea Tarazona, Jéssica Gil-Serna, Fernando Mateo, Belén Patiño and Eva María Mateo

Toxins 2025, 17(10), 487; https://doi.org/10.3390/toxins17100487 - 30 Sep 2025

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Abstract

Grapes are frequently contaminated by Aspergillus section Nigri fungi and ochratoxin A (OTA), with A. niger also capable of producing substantial fumonisin B₂ (FB₂) levels. Emerging evidence suggests that aflatoxigenic fungi may eventually replace ochratoxigenic fungi in certain regions due [...] Read more.

Grapes are frequently contaminated by Aspergillus section Nigri fungi and ochratoxin A (OTA), with A. niger also capable of producing substantial fumonisin B₂ (FB₂) levels. Emerging evidence suggests that aflatoxigenic fungi may eventually replace ochratoxigenic fungi in certain regions due to better adaptation to changes in climatic conditions. However, research on the toxigenic fungal community and mycotoxins in grapes from organic vineyards remains limited. Research on Spanish conventional grapes is also deficient, with most of the available literature being outdated. The present study investigates the diversity of toxigenic fungi and the presence of mycotoxins in organically cultivated grape berries in Spain, which are renowned for their significant oenological tradition. This study employed species-specific PCR protocols for fungal characterization and optimized methods for the analysis of OTA, FB₂, and aflatoxin B₁ (AFB₁) by UPLC–ESI–MS/MS. The most prevalent species present were Aspergillus flavus, A. niger, A. parasiticus, A. steynii, A. carbonarius, and A. westerdijkiae (67.1%, 43.5%, 20.0%, 14.1%, 14.1%, and 11.8% of the samples, respectively). OTA was detected only in 16 samples (19%), averaging 0.48 ng/g and peaking at 0.7 ng/g, which were lower than previously reported for conventional grapes. There was no FB₂ or AFB₁ detected. This study is pioneering in its exploration of the occurrence of toxigenic mycobiota, beyond Nigri fungi, and subsequent potential for other serious mycotoxins to contaminate Spain’s organic grapes. Full article

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

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