Toxins

24 pages, 1087 KiB

Open AccessArticle

Transcriptomics as an Early Warning of Domoic Acid Exposure in Pacific Razor Clams (Siliqua patula)

by Lizabeth Bowen, Shannon Waters, Brenda Ballachey, Heather Coletti, Zachary Forster, Jie Li and Bradley Jenner

Toxins 2025, 17(4), 194; https://doi.org/10.3390/toxins17040194 - 11 Apr 2025

Abstract

: As oceans warm, harmful algal blooms (HABs) are expected to increase, including blooms of Pseudo-nitzschia, a diatom that produces domoic acid (DA), which is a potent neurotoxin. Regulatory limits for human consumption (0.075–0.1 mg/kg/day; acute exposure) exist for the Pacific razor [...] Read more.

: As oceans warm, harmful algal blooms (HABs) are expected to increase, including blooms of Pseudo-nitzschia, a diatom that produces domoic acid (DA), which is a potent neurotoxin. Regulatory limits for human consumption (0.075–0.1 mg/kg/day; acute exposure) exist for the Pacific razor clam; however, fisheries currently do not have regulatory limits for chronic low-level exposure to DA even though razor clams can retain DA for over a year after an algal bloom. For bivalves, exposure to marine toxins may disrupt important cellular processes, leading to concerns about effects on their overall health and potential population- and ecosystem-level impacts. Transcriptomics was used to identify differentially expressed genes in razor clams (N = 30) from Long Beach, WA, collected prior to, during, and after a DA-producing bloom. Differentially expressed genes were identified that may indicate exposure of razor clams to DA, including clams with tissue DA concentrations that fall below regulatory limits for human consumption. Targeting these genes in real-time PCR assays may provide an early warning system for routine monitoring of DA in clams. Our results suggest DA exposure is associated with physiological responses ranging from decreased immune function to the potential disruption of cell communication, including retinoic acid catabolic processes, cell adhesion, collagen fibril organization, and immune effector processes. This work may also allow us to examine potential drivers of population-level change and whether chronic lower-level exposure to DA negatively impacts razor clam function, consequently affecting individual and population health. Full article

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

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Graphical abstract

17 pages, 1736 KiB

Open AccessArticle

Electrical Cell Impedance Sensing (ECIS): Feasibility of a Novel In Vitro Approach to Studying Venom Toxicity and Potential Therapeutics

by Abhinandan Choudhury, Kaitlin Linne, Tommaso C. Bulfone, Tanvir Hossain, Abu Ali Ibn Sina, Philip L. Bickler, Bryan G. Fry and Matthew R. Lewin

Toxins 2025, 17(4), 193; https://doi.org/10.3390/toxins17040193 - 11 Apr 2025

Abstract

Snakebite envenoming is often discussed in terms of lethality and limb loss, but local tissue injury and coagulotoxic effects of venom are significantly more common acute manifestations of snakebite envenoming (SBE). Local tissue injury and the hemorrhagic and coagulotoxic effects of venom are [...] Read more.

Snakebite envenoming is often discussed in terms of lethality and limb loss, but local tissue injury and coagulotoxic effects of venom are significantly more common acute manifestations of snakebite envenoming (SBE). Local tissue injury and the hemorrhagic and coagulotoxic effects of venom are challenging to study in live animals and can be ethically fraught due to animal welfare concerns such that attention to the 3Rs of animal welfare motivates the development of in vitro techniques in this arena. Herein, we tested the use of a wound-healing study technique known as Electric Cell-Substrate Impedance Sensing (ECIS) to assess populations of cultured cells exposed to venom with or without sPLA₂ and/or metalloprotease inhibitors (varespladib and marimastat, respectively). For comparison, the StarMax coagulation analyzer for coagulotoxicity was further used to evaluate the venoms and the neutralizing capabilities of the abovementioned direct toxin inhibitors (DTIs) against the same venoms examined using ECIS. Three viper and three elapid venoms that were examined for their effects on H1975 cells were Agkistrodon contortrix (Eastern Copperhead), Crotalus helleri (Southern Pacific Rattlesnake), and Vipera ammodytes (Horned Viper) and Naja atra (Chinese Cobra), Naja mossambica (Mozambique Spitting Cobra), and Naja nigricollis (Black-necked Spitting Cobra), respectively. The combination of cellular and coagulation techniques appears to usefully discriminate the in vitro capabilities and limitations of specific inhibitors to inhibit specific venom effects. This study suggests that ECIS with or without concomitant coagulation testing is a feasible method to generate reproducible, meaningful preclinical data and could be used with any type of cell line. Importantly, this approach is both quantitative and has the potential of reducing animal use and suffering during the evaluation of potential therapeutics. To further evaluate the potential of this method, rescue studies should be performed. Full article

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

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9 pages, 238 KiB

Open AccessReview

Safety of Onabotulinumtoxin-A for Chronic Migraine During Pregnancy and Breastfeeding: A Narrative Review

by Antonio Russo, Luigi Francesco Iannone, Ilaria Orologio, Veronica Rivi, Alberto Boccalini, Flavia Lo Castro, Marcello Silvestro and Simona Guerzoni

Toxins 2025, 17(4), 192; https://doi.org/10.3390/toxins17040192 - 11 Apr 2025

Abstract

Onabotulinumtoxin-A (onabotA) is a neurotoxin widely used for several indications, including chronic migraine (CM) preventive treatment, due to its well-demonstrated efficacy, tolerability, and safety. However, onabotA safety during pregnancy and breastfeeding remains unclear, as these populations are typically excluded from clinical trials. The [...] Read more.

Onabotulinumtoxin-A (onabotA) is a neurotoxin widely used for several indications, including chronic migraine (CM) preventive treatment, due to its well-demonstrated efficacy, tolerability, and safety. However, onabotA safety during pregnancy and breastfeeding remains unclear, as these populations are typically excluded from clinical trials. The action of onabotA starts locally at the injection sites, modulating the pain pathway with minimal systemic absorption, which theoretically minimizes risks to the fetus or breastfeeding infant. Preclinical studies demonstrate that onabotA does not distribute systemically in significant amounts after administration, although adverse fetal outcomes in rats and rabbits were reported when injected at high doses. Limited human data suggest that onabotA exposure during pregnancy may not be associated with major malformations or significant adverse outcomes for the fetus, especially when used at therapeutic doses for migraine prevention during the first trimester or earlier. Data on breastfeeding are even scarcer but indicate a low likelihood of drug transfer into breast milk. This narrative review highlights the available evidence on the use of onabotA in pregnancy and breastfeeding women, including real-word evidence, with a focus on the use for CM. Full article

3 pages, 176 KiB

Open AccessReply

Reply to Trosch et al. Comment on “Dashtipour et al. Dysphagia and Muscle Weakness Secondary to Botulinum Toxin Type A Treatment of Cervical Dystonia: A Drug Class Analysis of Prescribing Information. Toxins 2024, 16, 442”

by Khashayar Dashtipour, Han S. Lee, Aaron Ellenbogen, Rashid Kazerooni, Todd M. Gross, David A. Hollander and Conor J. Gallagher

Toxins 2025, 17(4), 191; https://doi.org/10.3390/toxins17040191 - 11 Apr 2025

Abstract

We thank Trosch et al [...] Full article

(This article belongs to the Section Bacterial Toxins)

3 pages, 162 KiB

Open AccessComment

Comment on Dashtipour et al. Dysphagia and Muscle Weakness Secondary to Botulinum Toxin Type A Treatment of Cervical Dystonia: A Drug Class Analysis of Prescribing Information. Toxins 2024, 16, 442

by Richard Trosch, Daniel Parreirinha and Susanne Proeschel

Toxins 2025, 17(4), 190; https://doi.org/10.3390/toxins17040190 - 11 Apr 2025

Abstract

We read with interest and some concern the article by Dashtipour and colleagues [...] Full article

11 pages, 1187 KiB

Open AccessCase Report

Poisoning from Alocasia × amazonica Roots: A Case Report

by Stanila Stoeva-Grigorova, Stela Dragomanova, Maya Radeva-Ilieva, Gabriela Kehayova, Simeonka Dimitrova, Simeon Marinov, Petko Marinov, Marieta Yovcheva, Diana Ivanova and Snezha Zlateva

Toxins 2025, 17(4), 189; https://doi.org/10.3390/toxins17040189 - 10 Apr 2025

Abstract

All parts of Alocasia × amazonica (A. amazonica, Araceae) pose a toxicological risk due to oxalate production. Ingestion of the plant extract may cause multi-organ damage and fatal outcomes. Given the rarity of poisoning cases, its toxicological profile and systemic effects [...] Read more.

All parts of Alocasia × amazonica (A. amazonica, Araceae) pose a toxicological risk due to oxalate production. Ingestion of the plant extract may cause multi-organ damage and fatal outcomes. Given the rarity of poisoning cases, its toxicological profile and systemic effects remain insufficiently characterized. This study aimed to investigate and report an appropriate approach to managing a patient intoxicated with A. amazonica (Araceae). A case of intentional self-poisoning with A. amazonica is presented. The patient, a 63-year-old woman, ingested approximately 200–300 mL of liquid prepared from the grated root of the plant. The initial clinical presentation involved localized injuries to the oral cavity and gastrointestinal tract, including severe pain, hoarseness, aphonia, dysphagia, mucosal erosions, and necrosis. Additional symptoms included hematinic vomiting, hemorrhagic diarrhea, and abdominal discomfort. These superficial and mucosal lesions resolved without the development of adhesions. Systemic effects comprised impaired consciousness indicative of encephalopathy, early metabolic acidosis, pulmonary edema with acute respiratory insufficiency, mild liver dysfunction, and hematuria. The therapeutic protocol for oral poisoning management was appropriate, leading to the patient’s discharge after 20 days of hospitalization. Full article

(This article belongs to the Special Issue Plant Toxin Emergency)

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16 pages, 6122 KiB

Open AccessCommunication

Assessing the Utility of Broad-Acting Inhibitors as Therapeutics in Diverse Venoms

by Raechel Kadler, Breanna Morrison and Angel Anne Yanagihara

Toxins 2025, 17(4), 188; https://doi.org/10.3390/toxins17040188 - 8 Apr 2025

Abstract

Examination of venom constituent bioactivities from diverse venomous animals shows certain highly conserved classes, including enzymes (e.g., phospholipases and metalloproteinases) and pore-forming proteins. While antivenoms targeting other unique and lethal venom components have proven to be life-saving, venom-enzyme-driven tissue damage and morbidity persists. [...] Read more.

Examination of venom constituent bioactivities from diverse venomous animals shows certain highly conserved classes, including enzymes (e.g., phospholipases and metalloproteinases) and pore-forming proteins. While antivenoms targeting other unique and lethal venom components have proven to be life-saving, venom-enzyme-driven tissue damage and morbidity persists. Broad-acting enzyme inhibitors demonstrate the potential to augment antivenom approaches. In this study, we investigate the potential utility of certain broad-acting inhibitors in cubozoa for the first time. Fluorogenic assays were used to determine the phospholipase A₂ (PLA₂) and matrix metalloproteinase (MMP) activity of the Hawaiian box jellyfish, Alatina alata, and this was compared to representative elapid, viper, and bee venoms. In vitro, evaluation of selected small-molecule inhibitors demonstrated the ability and feasibility of the broad-acting therapeutic doxycycline, which inhibited the PLA₂ and MMP activity of A. alata (approximately 50% reduction at 0.1 mM (95% CI 0.06–0.15) and 2.1 mM (95% CI 1.4–3.0), respectively), in addition to both snake venoms. Additionally, copper gluconate broadly inhibited the PLA₂ activity of bee, snake, and jellyfish venoms. While all venoms are complex mixtures of bioactive molecules, these studies demonstrate that targeting common class components with broad-acting inhibitors shows promise in clinical and preclinical management. Full article

(This article belongs to the Special Issue Venoms of Aquatic Organisms: Biochemistry, Evolution, and Future Perspectives)

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20 pages, 4316 KiB

Open AccessArticle

BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

by Jihong Li, Iman Mehdizadeh Gohari, Isabella Zhang and Bruce A. McClane

Toxins 2025, 17(4), 187; https://doi.org/10.3390/toxins17040187 - 8 Apr 2025

Abstract

By producing alpha toxin (PLC) and perfringolysin O (PFO), Clostridium perfringens type A strains are the most common cause of traumatic gas gangrene. C. perfringens cannot synthesize branched-chain amino acids (BCAAs), so BCAA transporters are essential for C. perfringens growth and survival. C. [...] Read more.

By producing alpha toxin (PLC) and perfringolysin O (PFO), Clostridium perfringens type A strains are the most common cause of traumatic gas gangrene. C. perfringens cannot synthesize branched-chain amino acids (BCAAs), so BCAA transporters are essential for C. perfringens growth and survival. C. perfringens type A strain ATCC3624 encodes the BrnQ1, BrnQ2, and BrnQ3 BCAA transporters. RT-PCR analyses showed that, with increasing culture time in TY broth, brnQ2 and brnQ3 expression levels remained stable but brnQ1 expression levels declined. Single null mutants unable to produce one of the BrnQ proteins grew and survived similarly as wild type. However, these mutants all showed altered PLC production, especially in the early culture stage, and those effects were reversible by complementation. Therefore, the presence of BrnQ proteins impacts toxin production levels, even though they are not necessary for growth. Interestingly, a triple mutant that was unable to produce any BrnQ protein also grew similarly as ATCC3624. Since BCAA uptake is essential for C. perfringens, this strain must produce another (still to be identified) BCAA transporter. Full article

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11 pages, 2044 KiB

Open AccessArticle

Pacific Ciguatoxin-1 (P-CTX-1) in a Moray eel (Gymnothorax javanicus) Responsible for Ciguatera in Khanh Hoa Province, Viet Nam

by Ha Viet Dao, Hy Ho Khanh Le, Ky Xuan Pham, Vy Bao Phan, Anh Phuong Nguyen, Thiet Thi Doan, Xuan-Vy Nguyen, Nhu-Thuy Nhat Nguyen, Xuan-Thuy Thi Nguyen, Tung Ngoc Nguyen, Jiajun Wu, Jingyi Zhu and Leo Lai Chan

Toxins 2025, 17(4), 186; https://doi.org/10.3390/toxins17040186 - 7 Apr 2025

Abstract

On 5 November 2020, a poisoning event involving four people by the consumption of moray eel occurred in Khanh Hoa Province, Viet Nam, with signs indicative of ciguatera. The remaining moray portion was confiscated for identification of causative species and responsible toxins. The [...] Read more.

On 5 November 2020, a poisoning event involving four people by the consumption of moray eel occurred in Khanh Hoa Province, Viet Nam, with signs indicative of ciguatera. The remaining moray portion was confiscated for identification of causative species and responsible toxins. The phylogenetic study based on COX1 identified the moray as Gymnothorax javanicus Bleeker (1859). Out of 17 marine lipophilic toxins (MLPs) that were analyzed using LC-MS/MS, only Pacific ciguatoxin-1 (P-CTX-1) was detected in the moray’s flesh at 1.30 ± 0.004 ng/g ww, while no toxin was found in the skin. The N2a assay’s ciguatoxicities in the skin and flesh were 0.69 ± 0.075 and 2.49 ± 0.216 ng P-CTX-1/g ww equivalent, respectively. In the N2a assay, the P-CTX-1 amount in the moray flesh was 1.9 times greater than that determined by LC-MS/MS, indicating the presence of additional sodium channel activators or a matrix effect. The P-CTX-1 amount in the moray flesh was at a level that generates major ciguatera poisoning (CP) symptoms in humans (1.0 ng/g P-CTX-1), makes sense given that four consumers experienced the onset of poisoning symptoms. This study is significant for the management of seafood safety since it is the first scientific report on the species and toxin in a moray causing ciguatera in Viet Nam. Full article

(This article belongs to the Collection Ciguatoxin)

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15 pages, 3553 KiB

Open AccessArticle

Bite First, Bleed Later: How Philippine Trimeresurus Pit Viper Venoms Hijack Blood Clotting

by Daniel Albert E. Castillo, Lorenzo Seneci, Abhinandan Chowdhury, Marilyn G. Rimando and Bryan G. Fry

Toxins 2025, 17(4), 185; https://doi.org/10.3390/toxins17040185 - 7 Apr 2025

Abstract

The Philippines has a high diversity of venomous snake species, but there is minimal information on their envenomation effects. This is evidenced by the small number of case reports, the poor reporting of envenomation cases, and the absence of specific antivenoms apart from [...] Read more.

The Philippines has a high diversity of venomous snake species, but there is minimal information on their envenomation effects. This is evidenced by the small number of case reports, the poor reporting of envenomation cases, and the absence of specific antivenoms apart from one against the Philippine cobra (Naja philippinensis). This study sought to profile the action of selected Philippine pit viper venoms on blood coagulation and to investigate whether commercially available non-specific antivenoms can provide adequate protection against these venoms. Venom from the pit vipers Trimeresurus flavomaculatus and Trimeresurus mcgregori were subjected to coagulation assays, antivenom cross-neutralization tests, and thromboelastography. Venoms from both species were able to clot human plasma and isolated human fibrinogen. Consistent with pseudo-procoagulant/thrombin-like activity, the resulting fibrin clots were weak and transient, thereby contributing to net anticoagulation through the depletion of fibrinogen levels. Clotting factors fIXa and fXa were also inhibited by the venoms, further contributing to the net anticoagulant activity. Monovalent and polyvalent antivenoms from the Thai Red Cross Society were effective against both venoms, indicating cross-neutralization of venom toxins; the polyvalent antivenom was able to rescue fibrinogen clotting to a greater degree than the monovalent antivenom. Our findings highlight the coagulopathic effects of these pit viper venoms and suggest the utility of procuring the non-specific antivenoms for areas in the Philippines with a high risk for pit viper envenomation. Full article

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

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13 pages, 6584 KiB

Open AccessArticle

Protective Effect of Lipoic Acid on Oxidative Stress and Tissue Damage Induced by Aflatoxin B₁ in Young Laying Hens

by Yihong Chu, Huanbin Wang, Xinyu Xu, Yun Ji, Yiting Zhao, Qianqian Yu, Shahid Ali Rajput, Yi Xue and Desheng Qi

Toxins 2025, 17(4), 184; https://doi.org/10.3390/toxins17040184 - 6 Apr 2025

Abstract

The aim of this study is to investigate the alleviating effect of lipoic acid on oxidative stress and tissue damage induced by aflatoxin B₁ (AFB₁) in young laying hens. The experiment was divided into a control group, an AFB₁ [...] Read more.

The aim of this study is to investigate the alleviating effect of lipoic acid on oxidative stress and tissue damage induced by aflatoxin B₁ (AFB₁) in young laying hens. The experiment was divided into a control group, an AFB₁ group, and three lipoic acid treatment groups. The AFB₁ group and three lipoic acid treatment groups were given diets supplemented with 90 μg/kg of AFB₁. The additional amounts of lipoic acid were 20, 100, and 500 mg/kg, respectively, with a feeding period of 4 weeks. The experimental results showed that AFB₁ significantly increased the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and malondialdehyde (MDA) in the serum and significantly decreased the levels of total protein (TP), albumin (ALB), total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) (p < 0.05). In addition, AFB₁ damaged the structure of the liver, spleen, and ovarian tissues. Lipoic acid reduced the levels of ALT, AST, ALP, and MDA in the serum and increased the levels of TP, ALB, T-SOD, GSH-Px, and CAT (p < 0.05). Meanwhile, lipoic acid also protected mitochondrial structure and alleviated liver, spleen, and ovarian tissue damage caused by AFB₁. In summary, lipoic acid can alleviate oxidative stress and tissue damage caused by AFB₁ in young laying hens. Full article

(This article belongs to the Special Issue Mycotoxin Contamination in Animal Nutrition and Its Mitigation Strategies)

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17 pages, 2538 KiB

Open AccessArticle

Insights into the Protein–Lipid Interaction of Perivitellin-2, an Unusual Snail Pore-Forming Toxin

by Romina F. Vázquez, M. Antonieta Daza Millone, Matías L. Giglio, Tabata R. Brola, Sabina M. Maté and Horacio Heras

Toxins 2025, 17(4), 183; https://doi.org/10.3390/toxins17040183 - 6 Apr 2025

Abstract

The perivitellin-2 (PV2) from snails is an unusual neuro and enterotoxin comprising a pore-forming domain of the Membrane Attack Complex and Perforin Family (MACPF) linked to a lectin. While both domains have membrane binding capabilities, PV2’s mechanism of action remains unclear. We studied [...] Read more.

The perivitellin-2 (PV2) from snails is an unusual neuro and enterotoxin comprising a pore-forming domain of the Membrane Attack Complex and Perforin Family (MACPF) linked to a lectin. While both domains have membrane binding capabilities, PV2’s mechanism of action remains unclear. We studied the apple snail Pomacea maculata PV2’s (PmPV2’s) interaction with lipid membranes using various biophysical and cell biology approaches. In vitro studies showed that PmPV2 toxicity decreased when cholesterol (Chol) was diminished from enterocyte cell membranes. Chol enhanced PmPV2 association with phosphatidylcholine membranes but did not induce pore formation. In contrast, using rat brain lipid models, rich in glycolipids, PmPV2 exhibited high affinity and induced vesicle permeabilization. Negative stain electron microscopy and atomic force microscopy confirmed the formation of pore-like structures in brain lipid vesicles. Our findings suggest that Chol is a necessary lipid component and point to PmPV2–glycolipid interactions as potential activators critical to triggering PmPV2’s pore-forming activity, providing insights into this novel toxin’s mechanism. Full article

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

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23 pages, 6254 KiB

Open AccessArticle

Computational Immunogenetic Analysis of Botulinum Toxin A Immunogenicity and HLA Gene Haplotypes: New Insights

by Eqram Rahman, Parinitha Rao, Munim Ahmed, William Richard Webb and Jean D. A. Carruthers

Toxins 2025, 17(4), 182; https://doi.org/10.3390/toxins17040182 - 6 Apr 2025

Abstract

Botulinum toxin A (BoNT-A) is widely used in both therapeutic and aesthetic settings; however, the formation of neutralizing antibodies (NAbs) remains a critical concern, leading to treatment failure. Immunogenic responses are known to vary between individuals due to HLA polymorphisms. Although some claim [...] Read more.

Botulinum toxin A (BoNT-A) is widely used in both therapeutic and aesthetic settings; however, the formation of neutralizing antibodies (NAbs) remains a critical concern, leading to treatment failure. Immunogenic responses are known to vary between individuals due to HLA polymorphisms. Although some claim that neurotoxin-associated proteins (NAPs) shield BoNT-A from immune detection or are themselves immunogenic, there is limited molecular evidence supporting either view. This study applies computational immunogenetics to explore BoNT-A immunogenicity, focusing on HLA binding and the influence of accessory proteins. Epitope mapping, molecular docking, and HLA binding predictions were used to evaluate interactions between BoNT-A epitopes and selected class II HLA alleles (HLA-DQA1*01:02, HLA-DQA1*03:03, HLA-DQB1*06:04, HLA-DQB1*03:01, and HLA-DRB1*15:01). To assess the potential immunomodulatory role of NAPs, molecular dynamics (MD) simulations, solvent-accessible surface area (SASA) analysis, and electrostatic potential mapping were also conducted. Key epitopes—L11, N25, and C10—showed strong binding affinities to HLA-DQA1*01:02, HLA-DQB1*06:04, and HLA-DQA1*03:03, indicating a potential immunodominant role. NAPs did not obstruct these epitopes but slightly increased their exposure and appeared to stabilize the toxin structure. Electrostatic mapping and binding free energy calculations suggested no significant immunogenic shift in the presence of NAPs. BoNT-A immunogenicity appears to be influenced by HLA allele variability, reinforcing the value of patient-specific genetic profiling. The presumed immunogenic role of NAPs remains unsubstantiated at the molecular level, underscoring the need for evidence-based evaluation over commercial rhetoric. While these findings provide valuable molecular insight, it is important to acknowledge that they are derived entirely from in silico analyses. As such, experimental validation remains essential to confirm the immunological relevance of these predicted interactions. Nonetheless, this computational framework offers a rational basis for guiding future clinical research and the development of HLA-informed BoNT-A therapies. Full article

(This article belongs to the Section Bacterial Toxins)

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14 pages, 3245 KiB

Open AccessArticle

Toxin Production by Alternaria alternata in Black Spot Disease of Chrysanthemum morifolium ‘Fubai’: Accumulation of Altenuene and Tenuazonic Acid in Flowers

by Qingling Zhan, Lina Liu, Wenjie Li, Jingshan Lu, Jiafu Jiang, Fadi Chen, Ye Liu and Zhiyong Guan

Toxins 2025, 17(4), 181; https://doi.org/10.3390/toxins17040181 - 5 Apr 2025

Abstract

Alternaria species produce diverse secondary metabolites that act as critical virulence factors during plant pathogenesis. In cultivation areas of Chrysanthemum morifolium ‘Fubai’—a key cultivar for herbal tea—black spot disease caused by A. alternata manifests as necrotic leaf lesions progressing to wilting. Despite this [...] Read more.

Alternaria species produce diverse secondary metabolites that act as critical virulence factors during plant pathogenesis. In cultivation areas of Chrysanthemum morifolium ‘Fubai’—a key cultivar for herbal tea—black spot disease caused by A. alternata manifests as necrotic leaf lesions progressing to wilting. Despite this disease’s economic impact, information on its associated toxins is limited, and the types of toxins produced by the black spot pathogen of Chrysanthemum morifolium ‘Fubai’ in particular remain unclear. Furthermore, whether toxins are present in the flowers when the leaves show symptoms is uncertain, but their inflorescence is not visibly infected. Using two previously characterized A. alternata strains (F16/F20) isolated from ‘Fubai’ in earlier studies, we demonstrated the concomitant production of altenuene (ALT) and tenuazonic acid (TeA) in both strains, with strain-specific yield variations (F20 TeA: 342.16 µg/mL vs. F16: 21.84 µg/mL; ALT: 0.28 µg/mL vs. 0.90 µg/mL). Time-course monitoring revealed coordinated accumulation of both toxins in inoculated petals, reaching 18.07 μg/g ALT and 2.59 µg/g TeA by day 9. Notably, these two toxins were detected only in flower samples from black spot-infected plants, and their concentrations correlated closely with disease severity in the leaves. Moreover, although the inflorescences did not display symptoms, both fresh and dried flowers retained detectable toxin levels. We established a technical system for the extraction and quantitative detection of the toxins ALT and TeA produced by the black spot pathogen in tea chrysanthemum. This work provides the first confirmation of ALT/TeA co-contamination in Chrysanthemum morifolium ‘Fubai’, revealing substantial dietary exposure risks through tea consumption. Our findings suggest that, from a food safety risk reduction perspective, integrated management strategies should be developed to minimize toxin contamination in tea chrysanthemum, including improved disease prevention measures and potential regulatory considerations. Full article

(This article belongs to the Section Mycotoxins)

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18 pages, 630 KiB

Open AccessStudy Protocol

Protocol and Demographics of the RELY-CD Study: Assessing Long-Term Clinical Response to Botulinum Neurotoxin in Cervical Dystonia

by Benjamin Waeschle, John-Ih Lee, Tristan Kölsche, Robin Jansen, Marta Banach, Stanislaw Ochudlo, Małgorzata Tyślerowicz, Piotr Sobolewski, Sara Sánchez Valiente, Eva López-Valdés, Pablo Mir, Silvia Jesús, Elena Ojeda-Lepe, Ewa Papuć, Pilar Sánchez Alonso, Gabriel Salazar, Georg Comes, Holger Stark and Philipp Albrecht

Toxins 2025, 17(4), 180; https://doi.org/10.3390/toxins17040180 - 5 Apr 2025

Abstract

The RELY-CD study investigated the long-term clinical response to botulinum neurotoxin type A in cervical dystonia within a multicenter, real-world setting. This retrospective study focused on patients treated with complex-free (incobotulinumtoxinA) and complex-containing (onabotulinumtoxinA and abobotulinumtoxinA) BoNT/A formulations over an up to 10-year [...] Read more.

The RELY-CD study investigated the long-term clinical response to botulinum neurotoxin type A in cervical dystonia within a multicenter, real-world setting. This retrospective study focused on patients treated with complex-free (incobotulinumtoxinA) and complex-containing (onabotulinumtoxinA and abobotulinumtoxinA) BoNT/A formulations over an up to 10-year period. The novel dose–effect parameter “DEff” was introduced to quantify the relationship between dose adjustments and clinical outcomes, enabling the identification of partial treatment failures. The primary endpoint was a comparison of a clinically meaningful worsening in DEff in treatment year 7 compared to year 2 between complex-free and complex-containing botulinum neurotoxin type A. The RELY-CD study provides unique insights into long-term treatment patterns, clinical resistance phenomena, and the implications of formulation differences on treatment outcomes, addressing a critical gap in the literature on real-world botulinum neurotoxin type A application. The study methodology, including the definition and calculation of the novel DEff, as well as clinical baseline characteristics, are presented. Full article

(This article belongs to the Special Issue Immunogenicity of Botulinum Toxin)

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15 pages, 1701 KiB

Open AccessReview

Are Aflatoxin Residues in Chicken Products a Real or Perceived Human Dietary Risk?

by Madalitso Chelenga, Limbikani Matumba, Muloongo C. Sitali, Bertha Kachala, Verson Nambuzi, Merning Mwenifumbo, Aggrey Pemba Gama, Mulunda Mwanza, Maurice Monjerezi and John F. Leslie

Toxins 2025, 17(4), 179; https://doi.org/10.3390/toxins17040179 - 4 Apr 2025

Abstract

Aflatoxin is a health threat to humans and domesticated animals. Chickens are often fed aflatoxin-contaminated grain and may retain some toxins in muscle, eggs, and other tissues. A critical food safety question is whether tissues from contaminated birds pose a threat to the [...] Read more.

Aflatoxin is a health threat to humans and domesticated animals. Chickens are often fed aflatoxin-contaminated grain and may retain some toxins in muscle, eggs, and other tissues. A critical food safety question is whether tissues from contaminated birds pose a threat to the humans that consume them. We evaluated literature published from 1984 to 2023 to determine the level of aflatoxin residues retained in chicken eggs, muscles, livers, gizzards, and hearts. In the studies evaluated (n = 33), ~8100 chickens in 334 trials were fed feed contaminated with 0.1–6400 µg/kg of aflatoxins for 7–180 days. There was a positive correlation between the level of feed contamination and residual aflatoxin concentrations (r² = 0.18, p < 0.05), but <1% of the aflatoxin in the feed carried over to edible broiler tissues. Only 0.6% of the trials reported >20 µg/kg of aflatoxin in the tissues, primarily in the muscle tissue, when the chickens were fed feed contaminated with >300 µg/kg of aflatoxins, which is above the US FDA maximum tolerable limit for components of poultry feeds. These composite results suggest a relatively low risk to public health from consuming chickens fed contaminated feed and a relatively high aflatoxin elimination mechanism in chickens that consume feed containing >300 µg/kg of aflatoxins. The data are consistent with chickens fed feed containing up to 500 µg/kg of aflatoxin being allowed in the human food chain without posing a significant health hazard. In reality, the maximum level of aflatoxin allowed in chicken feed will probably be limited by how much the birds can tolerate and still grow at a suitable rate without deformities rather than the risk that processed birds could present to human health. As chickens effectively act as an absorptive buffer for aflatoxin in contaminated feed, we expect that a contamination level that is acceptable for chicken growth performance is likely to be less than the amount that keeps chicken products safe for human consumption. Full article

(This article belongs to the Special Issue Aspergillus flavus and Aflatoxins (3rd Edition))

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15 pages, 4041 KiB

Open AccessArticle

Insights from Metabolomic and Transcriptomic Analyses into Sulforaphane’s Protective Mechanism Against Deoxynivalenol Toxicity via Spermine Regulation

by Yeyi Xiao, Jianliang Wu, Menke Feng, Jie Wang, Lele Qi, Chao Xu, Haifei Wang and Wenbin Bao

Toxins 2025, 17(4), 178; https://doi.org/10.3390/toxins17040178 - 3 Apr 2025

Abstract

Deoxynivalenol (DON) is a mycotoxin ubiquitously present in the environment. Emerging evidence demonstrated that sulforaphane (SFN) exerts potent protective effects against DON-triggered cytotoxicity through multimodal mechanisms. This study aimed to investigate the protective mechanism of SFN during DON exposure. Untargeted metabolomics of IPEC-J2 [...] Read more.

Deoxynivalenol (DON) is a mycotoxin ubiquitously present in the environment. Emerging evidence demonstrated that sulforaphane (SFN) exerts potent protective effects against DON-triggered cytotoxicity through multimodal mechanisms. This study aimed to investigate the protective mechanism of SFN during DON exposure. Untargeted metabolomics of IPEC-J2 cells revealed a total of 399 differential metabolites between the DON and control group and 365 differential metabolites between the SFN + DON and DON group. KEGG enrichment was performed to investigate the potential regulatory pathways. The transcriptome identified a total of 1839 differential expression genes (DEGs) between DON and SFN + DON groups. This result indicated that DON exposure and SFN treatment have a profound impact on cellular metabolism and genes. Integrated analysis of the transcriptome and metabolome showed that spermine was a potential biomarker for SFN treatment. SFN increased spermine abundance by regulating genes in glutathione, beta-alanine, and arginine and proline metabolism pathways. Functional experiments demonstrated that spermine alleviated DON-induced oxidative stress, as evidenced by increased cell viability, reduced ROS levels, restored mitochondrial membrane potential (ΔΨm), and normalized antioxidant enzyme activity. Moreover, spermine significantly decreased the cell apoptosis rate induced by DON, which suggested that spermine significantly alleviated the DON-induced cytotoxicity. Overall, these findings elucidated the protective role of SFN through spermine-related mechanisms against the toxicity of DON. Full article

(This article belongs to the Special Issue Alleviation of Mycotoxin-Induced Toxicity)

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16 pages, 3767 KiB

Open AccessArticle

Establishment and Comparison of Detection Methods for Ricin and Abrin Based on Their Depurination Activities

by Lina Dong, Tingting Liu, Jiaxin Li, Cen Wang, Jing Lv, Jing Wang, Jinglin Wang, Shan Gao, Lin Kang and Wenwen Xin

Toxins 2025, 17(4), 177; https://doi.org/10.3390/toxins17040177 - 3 Apr 2025

Abstract

Ricin (RT) and abrin (AT) are plant toxins extracted from Ricinus communis and Abrus precatorius, respectively, and both have N-glycosidase activity. The detection of these toxins is vital because of their accessibility and bioterrorism potential. While ricin can be effectively detected based [...] Read more.

Ricin (RT) and abrin (AT) are plant toxins extracted from Ricinus communis and Abrus precatorius, respectively, and both have N-glycosidase activity. The detection of these toxins is vital because of their accessibility and bioterrorism potential. While ricin can be effectively detected based on its depurination activity, only a few tests are available for detecting the depurination activity of abrin. Therefore, it is unclear whether they share the same optimal reaction substrate and conditions. Here, we established optimum depurination conditions for ricin and abrin, facilitating the in vitro detection of their depurination activity using high-performance liquid chromatography–tandem mass spectrometry. The parameters optimized were the reaction substrate, bovine serum albumin (BSA), buffer, pH, temperature, time, antibodies, and magnetic beads. Both toxins showed better depurination with single-stranded DNA. However, substrate length, adenine content, BSA concentration, buffer concentration, reaction temperature, and reaction time differed between the two toxins. The optimal conditions for ricin depurination involved a reaction in 1 mM ammonium acetate solution (0.5 μM DNA15A, 20 μg/mL BSA, and 1 mM Zn²⁺, with pH 4.0) at 55 °C for 1 h. The optimal conditions for abrin depurination involved a reaction in 1 mM ammonium citrate solution (0.2 μM DNA20A, 10 μg/mL BSA, 1 mM Mg²⁺, and 0.5 mM EDTA, with pH 4.0) at 45 °C for 2 h. After optimization, the limits of detection (LOD) for ricin and abrin were 0.506 ng/mL and 0.168 ng/mL, respectively. The detection time was also significantly reduced. Full article

(This article belongs to the Special Issue Novel Technologies for the Detection and Identification of Natural Toxins)

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6 pages, 194 KiB

Open AccessEditorial

Mycotoxins and Fungal Toxins: Current Status and Future Perspectives

by Jianhua Wang, Josefa Tolosa, Wenyu Wang and Xianli Yang

Toxins 2025, 17(4), 176; https://doi.org/10.3390/toxins17040176 - 3 Apr 2025

Abstract

Many toxigenic fungi are devastating pathogens of crop, fruit, and vegetable diseases worldwide [...] Full article

(This article belongs to the Special Issue Mycotoxins and Fungal Toxins: Current Status and Future Perspectives)

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