Toxins

19 pages, 1202 KB

Open AccessArticle

Antimicrobial Activity of Micrurus Venoms and Bioactive Films Functionalized with Purified L-Amino Acid Oxidase

by Vitelbina Núñez Rangel, Paola Rey-Suárez, Daniel Buitrago-Chinchilla, Laura Reyes-Méndez, Leidy Gómez-Sampedro, Alejandro Carmona-Jiménez, Mateo Rivillas-Ochoa and Adriana Muñoz-Bravo

Toxins 2026, 18(6), 240; https://doi.org/10.3390/toxins18060240 - 22 May 2026

Abstract

Phytopathogenic bacteria and fungi significantly reduce fruit and vegetable yields, resulting in substantial economic losses. Conventional management relies on synthetic agrochemicals; however, their intensive use poses risks to human health, environmental integrity, and biodiversity. Snake venoms have evolved under selective pressure, developing specialized [...] Read more.

Phytopathogenic bacteria and fungi significantly reduce fruit and vegetable yields, resulting in substantial economic losses. Conventional management relies on synthetic agrochemicals; however, their intensive use poses risks to human health, environmental integrity, and biodiversity. Snake venoms have evolved under selective pressure, developing specialized components with potent antimicrobial properties as part of a defense mechanism against prey-borne microorganisms. This study evaluated the inhibitory potential of Micrurus venoms against pathogens of agricultural interest and developed bioactive gelatin-based films incorporated with purified L-amino acid oxidases (LAAOs) as a novel biocontrol strategy. Venoms from M. ancoralis, M. mipartitus, and M. dumerilii exhibited significant growth inhibition against Xanthomonas and Fusarium strains. The primary active component was identified as LAAO through biological activity and mass spectrometry. Biofilms were formulated by incorporating M. ancoralis venom and its purified LAAO into a gelatin matrix. Physicochemical and microbiological characterization, alongside in situ assays on strawberries, demonstrated that the functionalized biofilms retained potent antimicrobial activity. Furthermore, LAAO incorporation did not significantly alter the physicochemical properties of the fruit but effectively extended shelf life by reducing weight loss and maintaining sensory appearance. These findings highlight the biotechnological potential of elapid venom components in the development of alternatives for phytopathogen control and active food packaging. Full article

(This article belongs to the Special Issue Venoms and Drugs)

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26 pages, 4280 KB

Open AccessArticle

Tracking Toxins: A Pilot Investigation of Cyanotoxins in North-Central Tennessee’s Surface Waters and Wells

by Kristi L. Hill, Andrea C. Jaegge, Devin M. Moore and Thomas D. Byl

Toxins 2026, 18(6), 239; https://doi.org/10.3390/toxins18060239 - 22 May 2026

Abstract

Cyanobacterial toxins (cyanotoxins) threaten aquatic ecosystems and human health, yet the factors influencing their production and distribution in freshwater remain unclear. In north-central Tennessee, nutrient-rich runoff from agricultural and urban areas, combined with a karst landscape that supports drinking and recreational water use, [...] Read more.

Cyanobacterial toxins (cyanotoxins) threaten aquatic ecosystems and human health, yet the factors influencing their production and distribution in freshwater remain unclear. In north-central Tennessee, nutrient-rich runoff from agricultural and urban areas, combined with a karst landscape that supports drinking and recreational water use, heightens the need to understand cyanotoxin behavior. To examine cyanotoxin patterns, the U.S. Geological Survey and the Tennessee Department of Environment and Conservation monitored 18 sites, including two wells under the influence of surface water, every two weeks from September 2022 to November 2024. At least one cyanotoxin was detected at all sites, with the highest concentrations in deep reservoirs and lower levels in shallow systems. Most detections occurred during summer and fall, aligning with high temperatures and rapid-onset drought. Statistical analysis indicated that increased specific conductivity and pH raised the likelihood of detecting total microcystin, likely resulting from drought conditions and nutrient-laden runoff. Additionally, dissolved microcystin showed an inverse relationship with Cumberland River water levels, and principal component analysis showed that Secchi depth, chlorophyll a, pH, temperature, and conductivity explained most water quality variability. These results help increase understanding of cyanotoxin distribution and associated water quality conditions during detections to guide future freshwater cyanotoxin monitoring studies. Full article

(This article belongs to the Special Issue Detection and Adsorption of Cyanotoxins in Waters)

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15 pages, 1016 KB

Open AccessReview

The Utility of the Electromyography and Ultrasound Guidance Combination for Botulinum Neurotoxin Injection: Focus on the Added Value of Electromyography

by Domenico Antonio Restivo, Mario Stampanoni Bassi, Rosario Marchese-Ragona, Giovanni Castelnovo, Angelo Alito, Demetrio Milardi, Stefano Masiero and Daniele Coraci

Toxins 2026, 18(6), 238; https://doi.org/10.3390/toxins18060238 - 22 May 2026

Abstract

The efficacy of botulinum neurotoxin (BoNT) is strongly dependent on its accurate delivery to hyperactive muscles and, ideally, to motor endplate regions. Although guidance techniques such as electromyography (EMG) and ultrasound (US) improve injection precision, each technique provides only partial information—either functional or [...] Read more.

The efficacy of botulinum neurotoxin (BoNT) is strongly dependent on its accurate delivery to hyperactive muscles and, ideally, to motor endplate regions. Although guidance techniques such as electromyography (EMG) and ultrasound (US) improve injection precision, each technique provides only partial information—either functional or anatomical. Integrating these techniques could enhance targeting accuracy, optimize dose distribution, and reduce off-target effects. A structured PubMed search was performed using terms related to BoNT, spasticity/dystonia, EMG, and US. Filters included clinical trials, randomized controlled trials, meta-analyses and reviews published within the last decade. Fifty-nine studies met the inclusion criteria. The publications were predominantly in neuroscience and rehabilitation journals. Only 17 studies reported combined EMG–US guidance. These focused mainly on stroke and cervical dystonia. While EMG-US integration is a promising strategy, we emphasize the added value of EMG guidance for US approaches, which is particularly important when treating complex neurological conditions involving complex, overlapping muscle activation patterns, or when targeting structures that are inaccessible to conventional imaging techniques. The EMG-US integrated approach is a promising strategy for optimizing BoNT therapy by combining structural visualization with real-time functional assessment. Despite its promising advantages in terms of accuracy and dose optimization, its clinical adoption is limited by a lack of high-quality evidence. Full article

(This article belongs to the Section Bacterial Toxins)

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26 pages, 1778 KB

Open AccessArticle

Phenotypic and Genotypic Characteristics of Staphylococcus aureus Nasal Strains Isolated from Students of the Pomeranian Medical University in Szczecin, Poland: A Cross-Sectional Study

by Paweł Kwiatkowski, Helena Masiuk, Agata Pruss, Stefania Giedrys-Kalemba, Piotr Baszuk, Iwona Wojciechowska-Koszko and Monika Sienkiewicz

Toxins 2026, 18(5), 237; https://doi.org/10.3390/toxins18050237 - 21 May 2026

Abstract

Staphylococcus aureus nasal carriage contributes to asymptomatic transmission in both community and healthcare settings. This study aimed to characterize S. aureus strains isolated from students of the Pomeranian Medical University in Szczecin, Poland, using phenotypic and genotypic methods. A total of 175 S. [...] Read more.

Staphylococcus aureus nasal carriage contributes to asymptomatic transmission in both community and healthcare settings. This study aimed to characterize S. aureus strains isolated from students of the Pomeranian Medical University in Szczecin, Poland, using phenotypic and genotypic methods. A total of 175 S. aureus strains were isolated from the nasal vestibules of 800 students between 2014 and 2015. Species identification and antimicrobial susceptibility testing were performed using standard microbiological methods, while virulence-associated genes and agr groups were analyzed using Single-PCR and Multiplex-PCR assays. Genotypic diversity was assessed by pulsed-field gel electrophoresis (PFGE). The prevalence of S. aureus nasal carriage among students was 21.9% and did not differ according to faculty or year of study. Most isolates (84.0%) were susceptible to all tested antibiotics, and no methicillin-resistant S. aureus (MRSA) strains were detected. All strains carried the hla gene, whereas hld and hlg were identified in 93.7% and 93.1% of isolates, respectively. In addition, the tst gene was detected in 22.3% of strains, while the lukS-PV/lukF-PV genes were identified in only one isolate (0.6%). The most prevalent enterotoxin genes were sep (17.1%) and sea (13.7%), whereas genes of the egc cluster, including seg, sei, and seo, were detected in 53.7% of isolates. Significant associations were observed between specific egc gene combinations and superantigen gene profiles, including increased frequencies of sec, sel, and tst genes (p < 0.001). The predominant agr type was agr-1 (49.7%), followed by agr-3 (28.6%) and agr-2 (20.0%). Strains carrying agr-1 more frequently harbored the g i m n o cluster as well as the sec, sel, and sep genes, whereas agr-3-positive isolates were significantly associated with the g i m o u and g i o u clusters and with the presence of tst, sea, and seh genes (p < 0.05). PFGE analysis demonstrated substantial genetic heterogeneity among the isolates, with no evidence of a predominant clonal lineage. These findings indicate a heterogeneous, non-epidemic population structure of S. aureus strains circulating among university students and highlight the considerable diversity and interrelationships of virulence-associated genetic profiles within this population. Full article

(This article belongs to the Special Issue Staphylococcus aureus Toxins：Presence and Detection in Human, Animals and Food)

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37 pages, 1571 KB

Open AccessReview

Biological Detoxification of Mycotoxins by Lactic Acid Bacteria: Safeguarding Food from Fungal Contaminants

by Nazia Tabassum, Minji Kim, Tae-Hee Kim, Du-Min Jo, Won-Kyo Jung, Young-Mog Kim and Fazlurrahman Khan

Toxins 2026, 18(5), 236; https://doi.org/10.3390/toxins18050236 - 20 May 2026

Abstract

Mycotoxins are one of the biggest threats to global food safety, public health, and economic stability. More than 400 mycotoxins have been found to be secondary metabolites of toxigenic fungi, mostly from the genera Aspergillus, Fusarium, Penicillium, and Alternaria. [...] Read more.

Mycotoxins are one of the biggest threats to global food safety, public health, and economic stability. More than 400 mycotoxins have been found to be secondary metabolites of toxigenic fungi, mostly from the genera Aspergillus, Fusarium, Penicillium, and Alternaria. Aflatoxins (AFs), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FBs), patulin (PAT), and T-2/HT-2 toxins are the most dangerous to the health of people and animals. Conventional physical and chemical decontamination methods are only partially effective and can reduce food quality, leave toxic residues, or be too expensive for smallholder food systems. Recent studies have shown that the application of lactic acid bacteria (LAB) as a biological detoxification method is a safe, cost-effective, and environmentally friendly option, and has a long history of safe use in fermented foods. Selected strains or taxonomic units have been granted GRAS status by the FDA or QPS (Qualified Presumption of Safety) status by EFSA. However, their use for mycotoxin detoxification still requires strain-level safety assessment and efficacy validation in the intended food matrix. There are several mechanisms by which LAB employ to reduce the bioavailability of mycotoxins in food systems: (i) physical adsorption via cell wall components such as peptidoglycan, teichoic acids, and exopolysaccharides; (ii) enzymatic biotransformation that may produce non-toxic or less-toxic metabolites, though the safety of degradation products requires case-by-case toxicological assessment; (iii) antifungal metabolite production that inhibits fungal growth and mycotoxin biosynthesis; and (iv) competitive exclusion of toxigenic fungi during fermentation. This comprehensive review examines the existing evidence on the detoxification of major food mycotoxins by LAB, with an emphasis on mechanisms, strain-specific efficacy, food-matrix applications, and factors that affect detoxification efficacy. Discussion has also been made of translating in vitro findings to in vivo settings and food-scale applications, alongside regulatory frameworks, current challenges, and future research directions. The review also suggests ways to combine LAB with new technologies, such as encapsulation, genetic engineering, and fermentation optimization, to make food systems safer by synergistically controlling mycotoxins. Full article

(This article belongs to the Special Issue Adverse Effects of Co-Contaminated Mycotoxins and Strategies for Their Mitigation)

51 pages, 6769 KB

Open AccessArticle

A Comprehensive Structural and Functional Analysis of Saccharomyces Killer Toxins

by Jack W. Creagh, Lily L. Givens, David C. Reetz, Sarah A. Coss, Rodolfo Bizarria, Jr., Siti Aisyah Alias, Mohammed Rizman-Idid, Jagdish S. Patel, Andre Rodrigues, F. Marty Ytreberg and Paul A. Rowley

Toxins 2026, 18(5), 235; https://doi.org/10.3390/toxins18050235 - 20 May 2026

Abstract

Antifungal killer toxins are cytotoxic proteins that have the potential to combat the growing threat of fungi to human health and agriculture. A lack of empirical tertiary structures has limited understanding of their mechanisms of action and their ability to target pathogens. In [...] Read more.

Antifungal killer toxins are cytotoxic proteins that have the potential to combat the growing threat of fungi to human health and agriculture. A lack of empirical tertiary structures has limited understanding of their mechanisms of action and their ability to target pathogens. In this study, AlphaFold and molecular dynamics simulations were used to generate tertiary structure models of all canonical Saccharomyces killer toxins and to place them in the context of historical empirical data. These models enabled the prediction of functional domains and posttranslational modifications, including proteolytic cleavage sites and disulfide bonds. They also revealed unexpected homology between Saccharomyces killer toxins, suggesting that all but K28 are likely ionophores. Structural homology to the well-studied killer toxins K1 and K2 enabled the prediction of the antifungal and immunity mechanisms of K1L, K21, K45, K74, and KHS. The understudied killer toxins Klus, KHR, and K62 were found to have homology to bacterial and plant toxins, including members of the aerolysin family and antifungal lectins. These structural similarities provide clues for the mechanisms of killer toxin carbohydrate binding, oligomerization, and membrane attack. This modeling approach will help guide the continued use of the model yeast S. cerevisiae to study killer toxins in the context of the wealth of functional data gathered in the decades since their first discovery. Full article

(This article belongs to the Special Issue Molecular Response of Hosts to Fungal Toxins)

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11 pages, 1783 KB

Open AccessArticle

New Furoisocoumarins with Phytotoxic Activity from the Fungus Aspergillus calidoustus VKM F-4916

by Tatiana V. Antipova, Vsevolod R. Dubovik, Anton N. Yurchenko, Olesya I. Zhuravleva, Valentina P. Zhelifonova, Elizaveta G. Lukina, Boris P. Baskunov, Oussama Abdelhamid Mammeri, Sergey N. Smirnov, Natalya E. Ivanushkina, Kirill V. Zaitsev, Qunfang Weng, Mikhail B. Vainshtein and Alexander O. Berestetskiy

Toxins 2026, 18(5), 234; https://doi.org/10.3390/toxins18050234 - 20 May 2026

Abstract

Aspergillus fungi are a source of low-molecular compounds of various structures possessing biological activities. We investigated the secondary metabolite profile of the soil fungus A. calidoustus VKM F-4916. The strain was found to synthesize new metabolites attributed to furoisocoumarins, which we named asperisocoumarin [...] Read more.

Aspergillus fungi are a source of low-molecular compounds of various structures possessing biological activities. We investigated the secondary metabolite profile of the soil fungus A. calidoustus VKM F-4916. The strain was found to synthesize new metabolites attributed to furoisocoumarins, which we named asperisocoumarin J and K, and a known siderophore desferritriacetylfusigen. The structure of asperisocoumarin J and K were determined by mass spectrometry and NMR spectroscopy. Asperisocoumarins J, K and desferritriacetylfusigen possessed a phytotoxicity, inhibiting the lettuce root growth. Sow thistle leaf and wheat leaf cuttings were sensitive to the action of asperisocoumarin J and K at a concentration of 5 mg/mL. Analysis of the structures of furoisocoumarins (asperisocoumarins J and K) using the online resource Pesti-DGI-Net showed that compounds had the physico-chemical properties favorable for pesticide development, in particular, fungicides and herbicides. An in-depth study of the phytotoxic properties of furoisocoumarins and their natural analogs is of interest in the context of the search for new herbicide compounds. Full article

(This article belongs to the Special Issue Fungal Phytotoxins: A Themed Issue in Honor of Prof. Antonio Evidente)

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10 pages, 3768 KB

Open AccessArticle

Differential Thermal Inactivation Enables Simultaneous Quantitation of Ricin and Abrin

by Woo-Hyeon Jeong

Toxins 2026, 18(5), 233; https://doi.org/10.3390/toxins18050233 - 19 May 2026

Abstract

Ricin and abrin are highly lethal Type II ribosome-inactivating proteins. They depurinate the same site of the 28S rRNA to inhibit protein synthesis. Consequently, standard molecular-level activity assays used to detect the toxic activity of ricin or abrin do not distinguish between the [...] Read more.

Ricin and abrin are highly lethal Type II ribosome-inactivating proteins. They depurinate the same site of the 28S rRNA to inhibit protein synthesis. Consequently, standard molecular-level activity assays used to detect the toxic activity of ricin or abrin do not distinguish between the two in mixed samples without prior physical separation or specially designed substrates. This study proposes a novel, cost-effective method to separately and simultaneously quantify the activities of ricin and abrin in mixtures by exploiting their distinct thermal stabilities. Thermal inactivation was used to demonstrate that heating samples at 80 °C for 5 min maximized the difference in their activities; while ricin retained most of its activity, abrin activity dropped to 20% after thermal treatment. This thermal treatment yielded 4 standard curves—ricin or abrin, thermally treated or not treated—in the 0.3 to 50 µg/mL range. By applying Cramer’s rule, the individual concentrations of active ricin and abrin in mixed samples were successfully calculated. However, this method should be used with a method detecting presence of ricin/abrin, to avoid unexpected reactivity due to contaminating RIPs. Full article

(This article belongs to the Collection Ribosome-Inactivating Proteins)

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13 pages, 271 KB

Open AccessPerspective

Beyond Wrinkle Efficacy: Toward a Broader Assessment of Longitudinal Compatibility in Routine Upper-Face Aesthetic BoNT-A

by Andrea Felice Armenti

Toxins 2026, 18(5), 232; https://doi.org/10.3390/toxins18050232 - 19 May 2026

Abstract

Botulinum toxin A (BoNT-A) is well established for the esthetic treatment of upper-face dynamic lines, and long-term adult studies suggest that repeated treatment may remain effective and acceptable over time in many patients. However, wrinkle efficacy and patient satisfaction do not by themselves [...] Read more.

Botulinum toxin A (BoNT-A) is well established for the esthetic treatment of upper-face dynamic lines, and long-term adult studies suggest that repeated treatment may remain effective and acceptable over time in many patients. However, wrinkle efficacy and patient satisfaction do not by themselves determine whether repeated treatment remains acceptable in broader morphologic, dynamic–expressive, and longitudinal terms. At the same time, the toxin’s pharmacology entails finite presynaptic blockade followed by active synaptic and architectural recovery, and imaging, histological, and neurophysiological studies indicate that repeated chemodenervation cannot be assumed to be biologically neutral at the muscle level. The resulting problem is not whether BoNT-A works, but whether current outcome frameworks are sufficient to judge repeated-treatment compatibility in full. Here, structural tolerance is proposed as a provisional clinical lens for an underdescribed boundary: whether repeated-treatment compatibility is fully captured by wrinkle reduction and patient acceptability alone. The paper organizes this problem across morphologic, dynamic–expressive, and longitudinal domains, outlines candidate warning signs, and develops a pragmatic tiered approach to assessment spanning routine care, structured clinical follow-up, and research-oriented evaluation. The aim is to support more complete longitudinal thinking in upper-face aesthetic BoNT-A, to clarify what current outcome frameworks remain unaddressed, and to identify priorities for future empirical study. Full article

(This article belongs to the Section Bacterial Toxins)

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12 pages, 1587 KB

Open AccessArticle

In Vivo Model of Short-Term Efficacy and Favorable Safety of Botulinum Toxin Type E Compared with Type A

by Jeong-Sun Nam, Daewon Yoon, Yujin Kim, Su-Young Kim, Jae-Young Kim, Yoonkyoung Cha, Joon Seok and Beom Joon Kim

Toxins 2026, 18(5), 231; https://doi.org/10.3390/toxins18050231 - 16 May 2026

Abstract

Botulinum toxin suppresses neurotransmitter release, thereby inhibiting muscle contraction and inducing flaccid paralysis. Botulinum toxin type A (BoNT/A) is widely used for neuromuscular blockade but, upon repeated administration, may cause long-lasting muscle atrophy, fibrosis, and inflammation. It is produced as a single peptide [...] Read more.

Botulinum toxin suppresses neurotransmitter release, thereby inhibiting muscle contraction and inducing flaccid paralysis. Botulinum toxin type A (BoNT/A) is widely used for neuromuscular blockade but, upon repeated administration, may cause long-lasting muscle atrophy, fibrosis, and inflammation. It is produced as a single peptide chain that becomes activated through cleavage into a heavy and light chain. BoNT/E, like BoNT/A, is produced as a single-chain polypeptide and requires cleavage to generate the active dichain form. Although BoNT/E is known to have a faster onset and shorter duration of action compared with BoNT/A, its efficacy and safety have not been thoroughly investigated. We compared BoNT/E and BoNT/A in SKH-1 hairless mice. Neuromuscular blockade, recovery pattern, and changes in muscle weight, volume, fiber size, fibrosis, mast cell infiltration, and diffusion to adjacent muscles were evaluated over time. BoNT/E induced maximal neuromuscular blockade on day 3 and fully recovered by day 35, whereas BoNT/A reached maximal effect on day 7 and showed only 20% recovery of the vehicle group by day 35. BoNT/E caused transient, dose-dependent reductions in muscle weight, volume, fiber size, and fibrosis, which largely normalized by day 35. In contrast, BoNT/A, administered at a dose of 0.5 U per injection site, induced persistent muscle atrophy, fibrosis, and significantly increased mast cell infiltration under the experimental conditions used in this study. Neither BoNT/E nor BoNT/A showed diffusion to adjacent muscles or changes in body weight. These findings suggest that BoNT/E provides rapid onset, short duration, and favorable safety, supporting its potential as an alternative therapeutic option for indications requiring temporary muscle relaxation with minimized long-term adverse effects. Full article

(This article belongs to the Section Bacterial Toxins)

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22 pages, 384 KB

Open AccessArticle

Multi-Mycotoxin Occurrence and Health Risk Assessment in Early Post-Harvest Durum Wheat, Barley, and Maize from Tunisia

by Khouloud Ben Hassouna, Jalila Ben Salah-Abbès, Samir Abbès, Ferjeni Zouidi, Mourad Jridi, Albert Sebastià, Noelia Pallarés and Houda Berrada

Toxins 2026, 18(5), 230; https://doi.org/10.3390/toxins18050230 - 16 May 2026

Abstract

The contamination of cereals by mycotoxins represents a major concern due to their harmful effects on human health and food quality. The current study investigated the occurrence of major mycotoxins (AFB1, AFB2, AFG1, AFG2, OTA, ENA, ENA1, ENB, and ENB1) in 158 raw [...] Read more.

The contamination of cereals by mycotoxins represents a major concern due to their harmful effects on human health and food quality. The current study investigated the occurrence of major mycotoxins (AFB1, AFB2, AFG1, AFG2, OTA, ENA, ENA1, ENB, and ENB1) in 158 raw cereal samples (durum wheat, barley, and maize) collected from two Tunisian regions: Beja (continental region) and Mahdia (coastal region). Mycotoxins were extracted using the QuEChERS method and quantified by UHPLC–MS/MS. Several mycotoxins were detected at high levels across all the cereals. In the Beja region, durum wheat was contaminated with AFB1, AFG1, ENA, ENA1, ENB, and ENB1, with ENB being the most frequent (70%). Mahdia durum wheat was contaminated only with ENA, ENA1, ENB, and ENB1, with ENB1 being the most prevalent (22.6%). Barley from both regions was contaminated only with ENs. The ENB was the most frequent (Beja 66%, Mahdia 28.6%). Maize from Beja was contaminated by OTA and ENs, with ENA1 being the most frequent (22.5%), while maize from Mahdia was contaminated by AFB1, AFB2, AFG2, and ENs (AFB1 was the most frequent, 35%). All wheat samples contaminated with AFG1 (6.55%) exceeded the European Union maximum limit for AFs in cereals (4 µg/kg). Similarly, maize samples contaminated with AFB1 (17.5%), AFB2 (10%), and AFG2 (2.5%) exceeded the EU maximum limit for aflatoxins in maize (10 µg/kg). Additionally, maize samples contaminated with OTA (5%) exceeded the EU maximum limit for OTA in unprocessed cereals (5 µg/kg). The co-occurrence of multiple mycotoxins was observed in all cereal types, with up to six different mycotoxins detected in a single sample. Dietary risk assessment revealed high EDIs of AFB1, AFG1, and ENs through the consumption of wheat and barley by Tunisian adults. The calculated MOE values for AFB1 and AFG1 in wheat were below 10,000 (MOE = 1190 for AFB1 and 2.5 for AFG1), suggesting a potential health concern associated with dietary exposure. Despite this potential risk, AFB1 and AFG1 were detected in only 3% and 7% of the analyzed samples, respectively. These results highlight the need for regular monitoring and the establishment of regulations to control mycotoxins in Tunisian cereals. Full article

(This article belongs to the Section Mycotoxins)

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19 pages, 2256 KB

Open AccessArticle

Efficient Biotransformation of Zearalenone in Acidic Food Matrices by Alkaline Enzyme–Inorganic Hybrid Nanoflower

by Ping Ding, Wenchao Liao, Chenyu Chen, Xincheng Chen, Chengfei Wang and Xiaoyang Li

Toxins 2026, 18(5), 229; https://doi.org/10.3390/toxins18050229 - 13 May 2026

Abstract

Zearalenone (ZEN) is a pervasive mycotoxin contaminating global food and feed. While enzymatic degradation offers a promising, specific, and eco-friendly strategy for mycotoxin mitigation, the biotransformation of ZEN within acidic food matrices remains challenging due to the intrinsically low activity of zearalenone lactonase [...] Read more.

Zearalenone (ZEN) is a pervasive mycotoxin contaminating global food and feed. While enzymatic degradation offers a promising, specific, and eco-friendly strategy for mycotoxin mitigation, the biotransformation of ZEN within acidic food matrices remains challenging due to the intrinsically low activity of zearalenone lactonase (ZENG). In this work, we synthesized a ZENG–hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂) hybrid nanoflower (CaNF) via biomineralization under alkaline conditions. Compared to free ZENG, the as-prepared biohybrid nanoflower exhibited markedly enhanced acid tolerance and catalytic activity, achieving a 12-fold increase in ZEN degradation efficiency at pH 5.0. Furthermore, the biohybrid nanoflower demonstrated robust performance in various acidic food matrices, including corn juice, wort, beer, and corn steep liquor. This study presents a powerful enzymatic tool for the efficient biotransformation of ZEN in acidic food-related systems. Full article

(This article belongs to the Section Mycotoxins)

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13 pages, 639 KB

Open AccessArticle

Early Intervention of Hemiplegic Shoulder Pain in the Context of Post-Stroke Shoulder Spasticity: A Canadian Cross-Sectional Survey

by Farris Kassam, Fraser MacRae, Linden Lechner, Heather Dow, Ève Boissonnault, Fiona Huang and Paul Winston

Toxins 2026, 18(5), 228; https://doi.org/10.3390/toxins18050228 - 12 May 2026

Abstract

Objectives: To investigate current Canadian physicians’ practice patterns of treating upper limb post-stroke spasticity (PSS) and hemiplegic shoulder pain (HSP) acutely after a stroke. In addition, by examining Canadian physicians’ diagnostic capabilities, time till treatment, minimum criteria to begin treatment, mechanisms of treatment, [...] Read more.

Objectives: To investigate current Canadian physicians’ practice patterns of treating upper limb post-stroke spasticity (PSS) and hemiplegic shoulder pain (HSP) acutely after a stroke. In addition, by examining Canadian physicians’ diagnostic capabilities, time till treatment, minimum criteria to begin treatment, mechanisms of treatment, targeting of muscles, and benefits and adverse effects of treatment, we aim to learn about areas of improvement to optimize PSS management for Canadians. Design: The present study was a cross-sectional survey, polling practicing Canadian physicians. Results: A total of 17 physicians completed the survey, all PM&R specialists, save one neurologist. Four provinces were represented in the responses. Participants had, on average, over ten years of experience managing post-stroke spasticity in outpatient and inpatient clinics. All 17 perform botulinum neurotoxin A (BoNT-A) injections for HSP associated with PSS. Most participants reported that they will begin BoNT-A treatments 2–3 weeks post-stroke, most commonly targeting the pectoralis major, subscapularis, and latissimus dorsi. Participants reported the mean median dosage they use as onabotulinum toxin A (169.12 units, SD = 73.70), incobotulinum toxin A (178.13 units, SD = 65.75), and abobotulinum toxin A (470.83 units, SD = 171.17). For injection guidance, participants responded that they use ultrasound for the largest percentage of their caseload, followed by electromyography, then electrical stimulation, then palpation. Very seldom did participants use palpation alone. Conclusions: From the limited sample included in analyses, the Canadian physicians respondents seem to be treating HSP and associated PSS with variable strategies. Further research is required to align dosages, targets, and guidance strategies as they vary considerably. Full article

(This article belongs to the Special Issue Botulinum Toxin: Advancing Treatments for Spasticity)

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21 pages, 2752 KB

Open AccessArticle

Nicotinamide Ameliorates Deoxynivalenol-Induced Injury in Renal Cells via Inhibiting PARP1 Hyperactivation and Restoring NAD⁺ Homeostasis

by Chao Chen, Yifan Qin, Zijun Luo, Peiqiang Mu, Jikai Wen and Yiqun Deng

Toxins 2026, 18(5), 227; https://doi.org/10.3390/toxins18050227 - 10 May 2026

Abstract

Deoxynivalenol (DON) is a globally prevalent mycotoxin that threatens food and feed safety via severe multi-organ toxicity. Previous studies indicate that DON induces cellular energy metabolism dysregulation by triggering oxidative stress and impairing mitochondrial function. During this process, nicotinamide adenine dinucleotide (NAD⁺ [...] Read more.

Deoxynivalenol (DON) is a globally prevalent mycotoxin that threatens food and feed safety via severe multi-organ toxicity. Previous studies indicate that DON induces cellular energy metabolism dysregulation by triggering oxidative stress and impairing mitochondrial function. During this process, nicotinamide adenine dinucleotide (NAD⁺), a central coenzyme in cellular energy metabolism, frequently exhibits significantly decreased intracellular levels or even complete depletion. However, the molecular mechanisms underlying the disruption of NAD⁺ homeostasis by DON exposure, as well as the development of targeted countermeasures, remain elusive. Using human embryonic kidney 293T (HEK293T) cells as an in vitro renal toxicity model, we dissected DON-induced NAD⁺ dysregulation and evaluated the protective potential of nicotinamide (NAM). DON caused significant NAD⁺ depletion in porcine serum (in vivo) and HEK293T cells (in vitro), which was confirmed as a key driver of cytotoxicity. Mechanistically, although DON binds and inhibits nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme of the NAD⁺ salvage pathway, neither NAMPT knockdown and overexpression nor nicotinamide mononucleotide (NMN) supplementation rescued DON-induced toxicity. Instead, DON dose-dependently activated poly(ADP-ribose) polymerase 1 (PARP1), the primary intracellular NAD⁺-consuming enzyme, to accelerate NAD⁺ depletion. PARP1 knockdown markedly attenuated DON-induced cytotoxicity, identifying PARP1 hyperactivation as the core toxic mechanism. NAM dose-dependently suppressed PARP1 activity, replenished NAD⁺ pools, and reversed cell injury. These findings establish PARP1-driven NAD⁺ depletion as an important mechanism of DON-induced renal toxicity, providing a promising intervention candidate for mitigating DON toxicity in food safety. Full article

(This article belongs to the Special Issue Adverse Effects of Co-Contaminated Mycotoxins and Strategies for Their Mitigation)

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21 pages, 2725 KB

Open AccessArticle

Response of Hemolytic and Photosynthetic Activity of Chattonella marina Complex Under Variable N:P Stoichiometry

by Xinyi Wang, Kehan Yi, Yongjun Jiang and Mengmeng Tong

Toxins 2026, 18(5), 226; https://doi.org/10.3390/toxins18050226 - 9 May 2026

Abstract

Chattonella marina is an ichthyotoxic, bloom-forming raphidophyte known for its hemolytic activity. However, the mechanisms by which nitrogen (N) and phosphorus (P) limitation influence this hemolytic toxicity remain poorly understood. In this study, both N and P limitation reduced growth, photosynthetic efficiency (F [...] Read more.

Chattonella marina is an ichthyotoxic, bloom-forming raphidophyte known for its hemolytic activity. However, the mechanisms by which nitrogen (N) and phosphorus (P) limitation influence this hemolytic toxicity remain poorly understood. In this study, both N and P limitation reduced growth, photosynthetic efficiency (F_v/F_m, Y_II, rETR_max), and the expression of nutrient-uptake, tetrapyrrole/chlorophyll biosynthesis genes. Nevertheless, the two nutrients produced opposite effects on toxicity: N limitation lowered hemolytic activity and ROS levels to near zero, whereas P limitation kept both relatively high, similar to nutrient-replete controls. The addition of the antioxidant NAC (N-Acetyl-L-cysteine) reduced hemolytic activity, confirming that ROS contributes to toxicity. Transcriptome data showed that under N limitation, genes for nitrogen uptake and initial reduction (NRT, NR, glnA) were upregulated, while downstream assimilation genes (nirA, GLT1) were downregulated. In contrast, under P limitation, all the nitrogen-metabolism-related genes (NRT, NR, glnA, nirA, GLT1) were downregulated. In the tetrapyrrole pathway, most genes were downregulated under both nutrient-limited conditions, except for HemD, suggesting a bottleneck that may result in the accumulation of porphyrin intermediates within the tetrapyrrole/chlorophyll biosynthesis pathway. Together, the secondary products derived primarily from the reaction of ROS with tetrapyrrole-based compounds appear to be the main contributors to hemolytic toxicity. Consequently, high levels of both ROS and porphyrin intermediates under P-limited conditions, as well as high ROS levels but low porphyrin intermediates under nutrient-sufficient conditions, may both contribute to the high hemolytic toxicity of C. marina. In contrast, under N limitation, despite the accumulation of porphyrin intermediates, the strong suppression of photosynthetic electron transport limits both ROS production and the synthesis of nitrogen-containing toxins, resulting in low hemolytic activity. These findings demonstrate that nutrient conditions regulate hemolytic activity in C. marina in a nutrient-specific manner. Full article

(This article belongs to the Special Issue Harmful Algal Toxins: Structure, Function, and Taxonomic Insights)

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25 pages, 8811 KB

Open AccessArticle

Plasma Metabolomics Reveals a Shared Metabolomic Profile in Experimental and Human Chronic Kidney Disease

by Søren H. Elsborg, Jasmine C. L. Atay, Johan Palmfeldt, Christian Daugaard Peters, Krista Dybtved Kjærgaard, Henricus A. M. Mutsaers and Rikke Nørregaard

Toxins 2026, 18(5), 225; https://doi.org/10.3390/toxins18050225 - 9 May 2026

Abstract

Chronic kidney disease (CKD) affects nearly 10% of the global population, yet diagnosis and disease monitoring still rely primarily on plasma creatinine. Because creatinine levels are strongly influenced by non–renal factors, such as age, sex, muscle mass, and diet, its accuracy as a [...] Read more.

Chronic kidney disease (CKD) affects nearly 10% of the global population, yet diagnosis and disease monitoring still rely primarily on plasma creatinine. Because creatinine levels are strongly influenced by non–renal factors, such as age, sex, muscle mass, and diet, its accuracy as a kidney function marker is limited. To identify plasma biomarkers that reflect kidney injury, we applied untargeted and targeted metabolomics in the adenine-induced CKD mouse model, a well-known tubular damage model, and validated the findings in plasma from patients with advanced CKD and healthy controls. We identified five metabolites that showed altered plasma levels in both experimental and human CKD, of which galactonic acid, pipecolic acid, and N-acetylneuraminic acid were significantly associated with measured glomerular filtration rate (GFR). As a proof-of-concept, we demonstrated that integrating these metabolites into a biomarker panel alongside creatinine could improve GFR estimation compared with creatinine alone. Our study introduces a promising metabolite-based biomarker panel that might enhance the accuracy of kidney function assessment and could potentially support diagnosis, risk stratification, and monitoring of disease progression; however, validation in a broader CKD cohort is needed. Full article

(This article belongs to the Section Uremic Toxins)

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17 pages, 543 KB

Open AccessArticle

Carry-Over Factor of Zearalenone in the Roof of the Third Ventricle of the Brain and Selected Skeletal Muscles During Low-Dose Zearalenone Mycotoxicosis in Prepubertal Gilts

by Magdalena Gajęcka, Łukasz Zielonka and Maciej T. Gajęcki

Toxins 2026, 18(5), 224; https://doi.org/10.3390/toxins18050224 - 8 May 2026

Abstract

The aim of this study was to determine whether exposure to low doses of zearalenone (ZEN) over a period of six weeks affects the values of the carry-over factor (COF) of ZEN in the roof of the third cerebral ventricle (RTCV) and in [...] Read more.

The aim of this study was to determine whether exposure to low doses of zearalenone (ZEN) over a period of six weeks affects the values of the carry-over factor (COF) of ZEN in the roof of the third cerebral ventricle (RTCV) and in selected skeletal muscles (longissimus and quadriceps) in prepubertal gilts. The study was conducted on 60 clinically healthy prepubertal gilts with an initial body weight (BW) of 14.5 ± 2 kg. Gilts were randomly assigned to a control group (group C; n = 15) and three experimental groups (ZEN5, ZEN10, and ZEN15; n = 15 each). Groups ZEN5, ZEN10, and ZEN15 were administered ZEN per os at doses of 5 µg/kg BW, 10 µg/kg BW, and 15 µg/kg BW, respectively. Group C animals were orally administered a placebo. Tissue samples (brain and skeletal muscles) were collected post-mortem for toxicological analyses on exposure days 7 (D1), 21 (D2), and 42 (D3). The concentrations of ZEN and its metabolites, α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL), were determined in the collected samples. All examined tissues contained the parent compound, but ZEN metabolites were not detected in any of the samples. The absence of ZEN metabolites may have resulted from a physiological deficit of estradiol (E₂) and, consequently, testosterone (T) and progesterone (P₄) in prepubertal gilts. Low-dose ZEN mycotoxicosis led to a persistent presence of ZEN in the RTCV (COF from 1 × 10⁻⁶ on D1 to 7 × 10⁻⁷ on D3) and somewhat lower ZEN levels in skeletal muscles (COF from 8 × 10⁻⁶ on D1 to 6 × 10⁻⁷ on D3). The presence of ZEN in the RTCV confirms that it crosses the blood–brain barrier and may therefore participate in the hormonal homeostasis of the brain. Full article

(This article belongs to the Section Mycotoxins)

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24 pages, 882 KB

Open AccessReview

Nutritional Factors Affecting Uremic Toxin Production

by Fanny Jouve, Christophe O. Soulage and Laetitia Koppe

Toxins 2026, 18(5), 223; https://doi.org/10.3390/toxins18050223 - 8 May 2026

Abstract

Chronic kidney disease (CKD) is characterized by persistent exposure to uremic toxins (UTs), many of which originate from gut microbial metabolism and contribute to renal, cardiovascular, and metabolic complications. Current evidence indicates that CKD is associated with dysbiosis and the enrichment of microbial [...] Read more.

Chronic kidney disease (CKD) is characterized by persistent exposure to uremic toxins (UTs), many of which originate from gut microbial metabolism and contribute to renal, cardiovascular, and metabolic complications. Current evidence indicates that CKD is associated with dysbiosis and the enrichment of microbial taxa carrying genes involved in UT precursor production. Diet is a major modulator of the gut microbiota and therefore represents a promising lever to reduce UT generation in synergy with current nephroprotective therapies. Beyond simple protein restriction, more specific dietary approaches, particularly plant-based low-protein diets, appear especially relevant. Additional factors, including amino acid composition, lipid quality, food processing, constipation, transit time, meal timing, and circadian rhythms, may also influence microbial metabolism and UT production. This review examines the role of nutrition in shaping the gut microbiota–UT–kidney axis and discusses how dietary modulation may support precision nutrition in the context of CKD. It also highlights future directions based on multidimensional phenotyping and robust biomarkers to capture interindividual variability, guide personalized interventions, and ultimately improve renal and cardiovascular outcomes in CKD. Full article

(This article belongs to the Special Issue New Insights in Uremic Toxins and Toxin-Related Outcomes in Chronic Kidney Disease)

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17 pages, 3039 KB

Open AccessArticle

Optimized SPE–UPLC–MS/MS Method for Sensitive Determination of Cereulide in Complex InfantFormula Matrices

by Zixiao Zhou, Ziyi Wang, Chundi Mu, Yan Qi, Jing Zhang, Xia Cui, Sai Fan, Jing Xiao and Rong Zhao

Toxins 2026, 18(5), 222; https://doi.org/10.3390/toxins18050222 - 8 May 2026

Abstract

Cereulide is a heat-stable cyclic depsipeptide toxin produced by Bacillus cereus and is responsible for foodborne emetic syndrome. Recent reports of Bacillus cereus contamination and cereulide occurrence in infant formula have raised increasing food safety concerns. Due to the immature immune and metabolic [...] Read more.

Cereulide is a heat-stable cyclic depsipeptide toxin produced by Bacillus cereus and is responsible for foodborne emetic syndrome. Recent reports of Bacillus cereus contamination and cereulide occurrence in infant formula have raised increasing food safety concerns. Due to the immature immune and metabolic systems of infants, exposure to cereulide through contaminated formula may lead to potential health risks. However, direct application of existing cereulide analytical methods to infant formula remains challenging because of the unique processing technologies, encapsulated nutrients, and variable matrix composition of this product category, which may hinder toxin release and cause significant matrix interference. In practical analysis, inter-laboratory comparisons revealed that existing methods exhibited relatively large deviations and insufficient sensitivity, making them not specifically optimized for infant formula matrices. The present study was motivated by the need for a matrix-specific, sensitive, and reliable analytical method for cereulide determination in infant formula. In this study, a method based on solid-phase extraction coupled with ultra-performance liquid chromatography–tandem mass spectrometry (SPE–UPLC–MS/MS) was developed and validated. To improve the applicability of cereulide analysis to infant formula, this method incorporates a hydration-assisted extraction step tailored to infant formula, which increased the detected cereulide response by approximately fourfold, together with optimized SPE clean-up and improved chromatographic conditions to reduce matrix effects and enhance quantitative reproducibility. The method showed good linearity (0.1–10 μg·L⁻¹, R² > 0.999), low values for limit of detection (LOD) (0.03 μg·kg⁻¹) and limit of quantification (LOQ) (0.1 μg·kg⁻¹), and acceptable recoveries (94.4–110.3%) with RSDs below 3.7%. The developed method was successfully applied to commercial infant formula samples, and cereulide-positive samples were identified. This method provides a reliable analytical tool for the monitoring of cereulide in infant formula and contributes to improved food safety surveillance and exposure risk assessment. Full article

(This article belongs to the Special Issue Detection and Risk Assessment of Microbial Toxins in Food)

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