Toxins

12 pages, 1332 KiB

Open AccessArticle

Real-World Insights into the Effectiveness and Tolerability of OnabotulinumtoxinA in Chronic Migraine: A Long-Term Evaluation of up to 11 Years

by Antonio Santoro, Andrea Fontana, Massimiliano Copetti, Anna Maria Miscio and Giuseppe d’Orsi

Toxins 2025, 17(4), 208; https://doi.org/10.3390/toxins17040208 - 21 Apr 2025

Viewed by 474

Abstract

Background: Chronic migraine (CM) is a debilitating neurological disorder that imposes substantial burdens on individuals and society, including diminished quality of life and increased healthcare utilization. While the efficacy of botulinum neurotoxin type A (BoNT-A) has been demonstrated in controlled trials, this longitudinal, [...] Read more.

Background: Chronic migraine (CM) is a debilitating neurological disorder that imposes substantial burdens on individuals and society, including diminished quality of life and increased healthcare utilization. While the efficacy of botulinum neurotoxin type A (BoNT-A) has been demonstrated in controlled trials, this longitudinal, real-world study offers unprecedented evidence of its long-term benefits, with patients followed for a median of 15 months (interquartile range: 6–36 months) and up to 11 years. Methods: This retrospective analysis included 579 patients diagnosed with CM who were newly treated with BoNT-A, according to the PREEMPT protocol, receiving injections every 12 weeks at doses of 155–195 units across 31–39 sites. Outcomes were assessed through changes in monthly headache days, frequency, symptomatic medication use, and migraine-related disability using Migraine Disability Assessment (MIDAS) scores up to 60 months from recruitment. Safety was evaluated by recording treatment-emergent adverse events (TEAEs), with a focus on long-term tolerability and subgroup variability. Results: Patients showed sustained improvements, with the mean number of monthly headache days decreasing from 22.7 to 5.5, and symptomatic medication use dropping from 33.4 to 3.7 mean doses at 60 months. Additionally, over 60% of patients improved from severe (MIDAS Grade IV) to minimal disability (MIDAS Grade I). Subgroup analysis revealed variability in response rates, emphasizing the need for personalized approaches. TEAEs were predominantly mild, with no new adverse events reported after 36 months, supporting the long-term safety of BoNT-A in real-world settings. Conclusions: This real-world study provides significant evidence for the long-term efficacy, safety, and tolerability of BoNT-A in the preventive treatment of CM. The findings highlight the importance of real-world data to account for patient variability and tailoring treatment strategies. Full article

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13 pages, 252 KiB

Open AccessPerspective

Expert Opinions on Best Practices for Overactive Bladder Management with onabotulinumtoxinA

by Karyn S. Eilber, Benjamin M. Brucker, Andrea Pezzella, Vincent Lucente, Kevin Benson and Michael J. Kennelly

Toxins 2025, 17(4), 207; https://doi.org/10.3390/toxins17040207 - 21 Apr 2025

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Abstract

OnabotulinumtoxinA is an FDA-approved treatment for adults with overactive bladder (OAB) who have an inadequate response to, or are intolerant of, oral pharmacotherapies including anticholinergics or beta-3 agonists. However, procedural practices of onabotulinumtoxinA intradetrusor injection vary among practitioners and can affect patient experience. [...] Read more.

OnabotulinumtoxinA is an FDA-approved treatment for adults with overactive bladder (OAB) who have an inadequate response to, or are intolerant of, oral pharmacotherapies including anticholinergics or beta-3 agonists. However, procedural practices of onabotulinumtoxinA intradetrusor injection vary among practitioners and can affect patient experience. To address this, a panel of six high-volume intravesical onabotulinumtoxinA providers with 100 years of combined experience convened to discuss the best office practices when treating patients with OAB. These key best practices include counseling patients on available OAB therapies, including onabotulinumtoxinA, at the initial consultation in accordance with established AUA and SUFU guidelines in a way that is easily understood. An office setting is preferred over a hospital or surgery center when performing the procedure. Staff involvement, from scheduling to post-procedure, is essential for establishing the relationships necessary to optimize patient experience and encourage compliance and retreatment. Experts generally recommend using a viscous lidocaine bladder instillation for an anesthetic 15 min prior to the reconstitution of onabotulinumtoxinA with 5 to 10 mL of normal saline. A range of one to 20 injection sites is acceptable, with a smaller number preferred. Starting in the lower bladder, experts recommend using a slower speed of injection to improve distribution and decrease patient discomfort. Subsequent treatments should be regularly scheduled at six-month intervals with the option of re-treating earlier if symptoms return, but no sooner than 12 weeks. For office intravesical onabotulinumtoxinA procedures, optimization of the patient experience by the physician and their staff, starting with the initial visit through the post-treatment follow-up, is key to long-term patient compliance. Full article

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

14 pages, 1890 KiB

Open AccessArticle

Toxicokinetics of a Single Oral Dose of Aflatoxin B₁ in Plasma, Feces, and Urine of Male Donkeys

by Yulong Feng, Min Li, Yunduo Zheng, Honglei Qu, Pengshuai Li, Boying Dong, Yantao Wang, Guangyuan Liu, Bin Jia and Qiugang Ma

Toxins 2025, 17(4), 206; https://doi.org/10.3390/toxins17040206 - 20 Apr 2025

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Abstract

Aflatoxin B₁ (AFB₁) is widely present in raw materials for food and feedstock, posing a significant threat to the health of humans and animals. This study explored the toxicokinetics of a single oral administration of AFB₁ at a dose [...] Read more.

Aflatoxin B₁ (AFB₁) is widely present in raw materials for food and feedstock, posing a significant threat to the health of humans and animals. This study explored the toxicokinetics of a single oral administration of AFB₁ at a dose of 100 µg·kg⁻¹ BW (body weight). Donkey blood samples were gathered at 0, 5, 10, 15, 20, 30, 45, and 60 min and at 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 6 h, 9 h, 12 h, 24 h, 48 h, 72 h, 96 h, and 120 h through jugular vein sampling needles at intervals. Fecal and urinary samples were collected at 0 h and every 6 h thereafter until 120 h. The concentrations of AFB₁ and AFM₁ in plasma, urine, and feces were quantitatively analyzed using LC-MS/MS. The maximum concentrations of AFB₁ and AFM₁ in plasma were 13.10 ± 6.35 µg·L⁻¹ and 0.72 ± 0.33 µg·L⁻¹, occurring at 1.38 ± 0.89 h and 2.25 ± 1.57 h after oral administration, respectively. The AFB₁ and AFM₁ elimination half-lives (T_1/2Elim) were 6.65 ± 2.84 h and 5.85 ± 3.00 h, respectively. The total clearances (CL) of AFB₁ and AFM₁ were 163 ± 52.2 L·kg⁻¹ BW⁻¹·h⁻¹ and 3210 ± 2450 L·kg⁻¹ BW⁻¹·h⁻¹, and the volumes of distribution (Vd) for AFB₁ were 1440 ± 417 L·kg⁻¹·BW and 22,400 ± 14,800 L·kg⁻¹·BW, respectively. In addition, the total amounts of AFB₁ and AFM₁ excreted over 120 h through urine and feces accounted for 3.38 ± 0.92% and 3.44 ± 1.45% of the total intake, respectively (calculated by material mass). Furthermore, the research showed that the absorption and metabolism of AFB₁ were rapid in male donkeys, with the tissue exhibiting a wide distribution and long duration. Full article

(This article belongs to the Section Mycotoxins)

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11 pages, 258 KiB

Open AccessArticle

Autonomic Dysfunction and Blood Pressure Variability in Botulinum Intoxication: A Prospective Observational Study from a Single-Center Italian Outbreak

by Giuseppe Miceli, Giuliano Cassataro, Vito Volpe, Emanuela Fertitta, Carmelinda Canale, Lucia Tomaiuolo, Melania Blasco, Mariagrazia Stella, Matteo Velardo and Maurizio Renda

Toxins 2025, 17(4), 205; https://doi.org/10.3390/toxins17040205 - 20 Apr 2025

Viewed by 354

Abstract

Botulinum neurotoxin (BoNT) intoxication is a rare but severe condition that is characterized by autonomic and neuromuscular dysfunction. This study aimed to evaluate autonomic impairment and blood pressure variability in patients with botulinum intoxication during an outbreak, compared to healthy controls, and to [...] Read more.

Botulinum neurotoxin (BoNT) intoxication is a rare but severe condition that is characterized by autonomic and neuromuscular dysfunction. This study aimed to evaluate autonomic impairment and blood pressure variability in patients with botulinum intoxication during an outbreak, compared to healthy controls, and to assess their progression over a six-month follow-up period. Methods: Twenty (n = 20) male patients diagnosed with BoNT intoxication and 34 age- and sex-matched healthy controls were enrolled. At baseline, all subjects underwent 24 h ambulatory blood pressure monitoring (ABPM), and clinostatic and orthostatic blood pressure measurements. Autonomic function parameters, including mean systolic blood pressure (SBP), mean diastolic blood pressure (DBP), SBP and DBP variability, SBP and DBP load, pulse pressure (PP), blood pressure variability ratio (BPVR), and morning surge, were analyzed. Follow-up assessments were conducted after six months. Results: Patients with botulinum intoxication exhibited significantly lower SBP, DBP, and blood pressure variability parameters compared to healthy controls. Orthostatic hypotension was present in 55% of patients at baseline, improving to 5% at follow-up. Respiratory failure occurred in 40% of cases, necessitating non-invasive ventilation in 35% and intubation in 20%. At six-month follow-up, mean SBP, DBP, heart rate, and blood pressure variability parameters increased significantly, indicating partial recovery of autonomic control. However, residual abnormalities in autonomic regulation persisted. Conclusions: BoNT intoxication leads to notable autonomic dysfunction, marked by impaired blood pressure regulation and a high prevalence of orthostatic hypotension. Although partial recovery occurs, long-term autonomic impairment persists, highlighting the necessity for ongoing cardiovascular monitoring and further research to accelerate autonomic recovery through targeted therapeutic interventions. Full article

(This article belongs to the Section Bacterial Toxins)

15 pages, 1265 KiB

Open AccessSystematic Review

Effectiveness of Bee Venom Injection for Parkinson’s Disease: A Systematic Review

by Hyein Jeong, Kyeong Han Kim and Seong-gyu Ko

Toxins 2025, 17(4), 204; https://doi.org/10.3390/toxins17040204 - 20 Apr 2025

Viewed by 502

Abstract

Parkinson’s disease (PD) is the second most common neurodegenerative disorder worldwide, affecting over 8.5 million people as of 2019. While standard pharmacological treatments help alleviate symptoms, their long-term use can lead to side effects such as dyskinesia. Bee venom acupuncture (BVA) involves the [...] Read more.

Parkinson’s disease (PD) is the second most common neurodegenerative disorder worldwide, affecting over 8.5 million people as of 2019. While standard pharmacological treatments help alleviate symptoms, their long-term use can lead to side effects such as dyskinesia. Bee venom acupuncture (BVA) involves the use of a natural toxin derived from bees that can be used for pain relief and treating neurological disorders. This study aimed to review the efficacy and safety of BVA for the treatment of PD. This review protocol was prospectively registered with PROSPERO (CRD420251000577). We searched eight databases in February 2025 and selected 12 studies involving 215 PD patients treated with BVA. Idiopathic Parkinson’s disease (IPD) is the most common diagnosis. The concentration and dosage per session ranged from 0.03 to 0.1 mg/mL and from 0.1 to 1.0 mL, respectively. Twenty-four different outcome measures were used, with the Unified PD Rating Scale employed in 91.7% of the studies. All studies reported improvements in outcomes. Mild adverse effects such as swelling and itching were noted in four studies (33.3%); however, no severe reactions such as anaphylactic shock occurred. These findings suggest that BVA has the potential for broader clinical applications in the treatment of PD. Full article

(This article belongs to the Special Issue Clinical Evidence for Therapeutic Effects and Safety of Animal Venoms)

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17 pages, 7461 KiB

Open AccessArticle

Apoptotic Effect of Combinations of T-2, HT-2, and Diacetoxyscirpenol on Human Jurkat T Cells

by Phattarawadee Wattanasuntorn, Saranya Poapolathep, Patchara Phuektes, Imourana Alassane-Kpembi, Johanna Fink-Gremmels, Isabelle P. Oswald and Amnart Poapolathep

Toxins 2025, 17(4), 203; https://doi.org/10.3390/toxins17040203 - 18 Apr 2025

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Abstract

Trichothecene type A mycotoxins, such as T-2, HT-2, and diacetoxyscirpenol (DAS), are known to induce cytotoxicity and apoptosis in different cell types. As all three Fusarium toxins may occur concomitantly in a given food or feed commodity, there is growing interest in the [...] Read more.

Trichothecene type A mycotoxins, such as T-2, HT-2, and diacetoxyscirpenol (DAS), are known to induce cytotoxicity and apoptosis in different cell types. As all three Fusarium toxins may occur concomitantly in a given food or feed commodity, there is growing interest in the effect of such mycotoxin mixtures. This study aimed to identify the toxic interactions among T-2, HT-2, and DAS in a human Jurkat cell model. As a first step, an MTT assay was used to assess cytotoxicity after 24 h of cell exposure to individual mycotoxins and their mixtures. The results were used to calculate the combination index (CI), which indicates the nature of the mycotoxin interactions. In Jurkat T cells, the toxicity ranking for the individual mycotoxins was T-2 > HT-2 > DAS. The CI values of the dual and triple mycotoxin combinations calculated from the results of the MTT and reactive oxygen species assays showed synergistic effects at low concentrations and an apparent antagonism at very high concentrations for all combinations. The additional cytometric analyses confirmed the synergistic effects, as expected, following co-exposure to the three tested trichothecenes. As the lower toxin concentrations investigated reflect natural contamination levels in food and feeds, the synergistic effects identified should be considered in risk characterization for trichothecene exposure in humans and animals. Full article

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

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12 pages, 239 KiB

Open AccessArticle

Efficacy and Predictive Factors of Intravesical Botulinum Toxin A Injection for Treating Neurogenic Detrusor Overactivity in Children: A Single-Center Retrospective Study

by Chun-Kai Hsu, Han-Yu Lin, Stephen Shei-Dei Yang, Wan-Ling Young and Shu-Yu Wu

Toxins 2025, 17(4), 202; https://doi.org/10.3390/toxins17040202 - 17 Apr 2025

Viewed by 405

Abstract

Neurogenic detrusor overactivity (NDO) is a complex condition associated with detrusor overactivity, reduced bladder compliance, and high intravesical pressures, potentially leading to urinary tract infections (UTIs) and renal impairment. This retrospective study evaluated the safety and potential efficacy of intravesical botulinum toxin A [...] Read more.

Neurogenic detrusor overactivity (NDO) is a complex condition associated with detrusor overactivity, reduced bladder compliance, and high intravesical pressures, potentially leading to urinary tract infections (UTIs) and renal impairment. This retrospective study evaluated the safety and potential efficacy of intravesical botulinum toxin A (BoNT/A) injections in children with NDO at a single institution. Eighteen pediatric patients (median age: 8.1 years) were followed for a median of 6.3 years. At follow-up, 77.8% achieved a global response assessment (GRA) score of ≥2. A statistically significant improvement was found in cystometric bladder capacity (p = 0.041), but it did not remain significant after Bonferroni correction, while other urodynamic trends were not statistically significant. Adverse events were infrequent, with only 11.8% experiencing mild febrile UTIs. While some patients with poorer baseline bladder conditions reported greater subjective improvement, no statistically significant predictors of success were identified. Overall, intravesical BoNT/A injection appears to be a safe and potentially effective option for managing pediatric NDO, though larger prospective studies are needed to confirm these findings. Full article

(This article belongs to the Special Issue Application of Botulinum Neurotoxin in Lower Urinary Tract Dysfunctions: Where Are We Now? II)

15 pages, 3663 KiB

Open AccessArticle

Trends in Mycotoxins Co-Occurrence in the Complete Feed for Farm Animals in Southern Romania During 2021–2024 Period

by Valeria Cristina Bulgaru, Mihail Alexandru Gras, Aglaia Popa, Gina Cecilia Pistol, Ionelia Taranu and Daniela Eliza Marin

Toxins 2025, 17(4), 201; https://doi.org/10.3390/toxins17040201 - 15 Apr 2025

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Abstract

Mycotoxins are common natural contaminants of crops and fruits, associated with negative effects on human and animal health. Currently, more than 300 mycotoxins have been identified, but data on their effects and their limits in feed and food are still inconsistent. The European [...] Read more.

Mycotoxins are common natural contaminants of crops and fruits, associated with negative effects on human and animal health. Currently, more than 300 mycotoxins have been identified, but data on their effects and their limits in feed and food are still inconsistent. The European Commission, by directive EC 574/2011, established regulations concerning the maximum limit allowed in farm animals’ feed for aflatoxins, but for all other mycotoxins there are only recommendations (EC 1319/2016) and there are no established limits. Considering their variety and toxic effects, but also the fact that not many details are yet known about the cumulative effects of co-contamination with various mycotoxins, it is necessary to monitor the evolution of their presence in animal feed. The aim of our study was to analyze for a four-year period (2021–2024) the concentrations of six mycotoxins (total aflatoxins-AFT, fumonisins-FB, deoxynivalenol-DON, zearalenone-ZEA, T2/HT2 and ochratoxin (A + B)-OTA), the most frequently encountered in the south area of Romania in poultry, piglets and pig’s complete feed. Our results showed that the maximum highest concentrations were 5.8 ppb for AFT, 4.7 ppm for FB, 1.9 ppm for DON, 62.8 ppb for ZEA, 32.1 ppb for T2/HT2 and 19.7 ppb for OTA irrespective of the type of feed. It should be noted that AFT and ZEA were identified in all samples during the entire monitored period, and the only mycotoxin that exceeded the guidance value was DON, for which the recommendation of 0.9 ppm for pig feed was exceeded. Recent studies demonstrated that sub-chronic and chronic exposure to low concentrations of mycotoxins and specially co-contamination is more common than acute exposure, being able to affect animal health over time by lowering the defense capacity, inducing inflammatory reactions and affecting intestinal health, which in the long term could have important economic consequences. Our survey study can provide important data showing the degree of contamination with mycotoxins in pig and poultry feed including the simultaneous presence of different mycotoxins in this complete feed. Full article

(This article belongs to the Section Mycotoxins)

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11 pages, 374 KiB

Open AccessArticle

The Effects of Botulinum Toxin A Injections on Patients with Radiogenic Lower Urinary Tract Symptoms

by Anke K. Jaekel, Ann-Christin Brüggemann, John Bitter, Franziska Knappe, Ruth Kirschner-Hermanns and Stephanie C. Knüpfer

Toxins 2025, 17(4), 200; https://doi.org/10.3390/toxins17040200 - 15 Apr 2025

Viewed by 390

Abstract

Botulinum toxin A (BTX-A) injection into the detrusor vesicae is an established therapy for neurogenic lower urinary tract dysfunction as well as idiopathic overactive bladder. Pelvic radiotherapy causes comparable lower urinary tract symptoms (LUTS) in a third of radiated patients. Little is known [...] Read more.

Botulinum toxin A (BTX-A) injection into the detrusor vesicae is an established therapy for neurogenic lower urinary tract dysfunction as well as idiopathic overactive bladder. Pelvic radiotherapy causes comparable lower urinary tract symptoms (LUTS) in a third of radiated patients. Little is known about the effects of BTX-A injections into the detrusor vesicae in the management of radiogenic LUTS. Our aim was to assess the effect of BTX A injections on these symptoms and related quality of life. Material and Methods: In total, 28 patients with BTX-A injections for radiogenic LUTS were assessed retrospectively. We analyzed symptoms recorded in bladder diaries, the results of quality-of-life questionnaires (ICIQ-LUTSqol), and urodynamic studies (UDS) before and after BTX-A injections. Results: A significant reduction in daily micturition frequency, nocturia, and pad consumption was demonstrated in the overall cohort and in gender-related subgroup analysis. There was a significant decrease in the ICIQ-LUTSqol independent of gender or BTX-A units. For UDS maximum cystometric bladder capacity (188.0 vs. 258.2 mL, p = 0.043), micturition volume (138.2 vs. 216.7 mL, p = 0.018), and first desire to void (98.2 vs. 171.2 mL, p = 0.042) was significantly improved. No side effects of the toxin injection or urinary retention were observed. Conclusions: Intradetrusor injection therapy with BTX-A could represent a safe and effective therapeutic option for radiogenic LUTS with increasing quality of life, reductions in symptoms, and the improvement of urodynamic parameters. Full article

(This article belongs to the Special Issue Application of Botulinum Neurotoxin in Lower Urinary Tract Dysfunctions: Where Are We Now? II)

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14 pages, 871 KiB

Open AccessArticle

Variation in Subtypes of Obsessive-Compulsive Traits in Migraine Patients Undergoing Onabotulinum Toxin A Therapy

by Giovanna Viticchi, Lorenzo Falsetti, Chiara Di Felice, Gioacchino De Vanna, Sergio Salvemini, Marco Bartolini, Gianluca Moroncini and Mauro Silvestrini

Toxins 2025, 17(4), 199; https://doi.org/10.3390/toxins17040199 - 14 Apr 2025

Viewed by 432

Abstract

Background: Patients with chronic migraine (CM) associated with medication overuse headache (MOH) often exhibit concomitant psychiatric traits including obsessive-compulsive disorder (OCD). Limited data exist on the impact of migraine therapies on these traits. This study aimed to analyse the influence of onabotulinum toxin [...] Read more.

Background: Patients with chronic migraine (CM) associated with medication overuse headache (MOH) often exhibit concomitant psychiatric traits including obsessive-compulsive disorder (OCD). Limited data exist on the impact of migraine therapies on these traits. This study aimed to analyse the influence of onabotulinum toxin A (OBT-A) on OCD in CM + MOH patients. Methods: All CM + MOH patients attending the AOU-Marche Headache Centre and treated with OBT-A over a 9-month period were prospectively analysed. At baseline and every three months, patients completed several questionnaires, including the Obsessive-Compulsive Inventory-Revised (OCI-R), to assess the presence of OCD and its subscales. Results: Thirty patients were enrolled. Repeated measures tests revealed a statistically significant decrease from T0 to T3 in the OCI-R score (p = 0.017) and among the different subscales, specifically the checking score (p = 0.029). The MIDAS (migraine disability assessment score) and HIT-6 (headache impact test) scores exhibited a statistically significant reduction from T0 to T3 (p < 0.0001), similar to the decrease in monthly migraine days and symptomatic medication intake. Conclusions: Patients treated with OBT-A showed significant improvement in OCD, particularly in subscales assessing somatic and aggressive obsessions as well as control compulsions. Several patients transitioned from a CM + MOH condition to an episodic form without drug abuse. The potential impact of OBT-A on psychiatric symptoms warrants further consideration to improve patient management strategies. Full article

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

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18 pages, 1858 KiB

Open AccessArticle

A Sting Operation: Risk Assessment and Venom Expenditure by Arizona Bark Scorpions (Centruroides sculpturatus) in a Defensive Context

by Lindsay A. Marston, Gerad A. Fox, Kim Y. Hung, Shannon J. Delo and William K. Hayes

Toxins 2025, 17(4), 198; https://doi.org/10.3390/toxins17040198 - 13 Apr 2025

Viewed by 438

Abstract

Scorpion antipredator behavior incorporates risk assessment that informs decision-making and venom usage. We quantified antipredator behaviors of the clinically significant Arizona bark scorpion (Centruroides sculpturatus) in their natural environment using exposure to two stimuli: a freshly thawed laboratory mouse (Mus [...] Read more.

Scorpion antipredator behavior incorporates risk assessment that informs decision-making and venom usage. We quantified antipredator behaviors of the clinically significant Arizona bark scorpion (Centruroides sculpturatus) in their natural environment using exposure to two stimuli: a freshly thawed laboratory mouse (Mus musculus) and a membrane-covered glass beaker. We videotaped and compared envenomation behaviors between sexes (females, gravid females, and males), across sizes, and between animal orientations (on vertical or horizontal substrates). Results failed to show consistent support for any of our four hypotheses. Females (especially gravid females) were no more likely than males to exhibit higher levels of stinging and venom expenditure. Scorpions on horizontal surfaces compared to those on vertical surfaces, and larger scorpions compared to smaller ones, were likewise no more likely to exhibit higher levels of responsiveness. Mice were more likely to be stung than the membrane-covered beaker, but with fewer and briefer stings, suggesting the scorpions did not attempt to deliver more venom into the mice. Thus, we discerned no clear patterns in risk assessment, stinging, and venom use associated with sex, substrate orientation, body size, or threat stimuli. These findings contrasted with those of several prior laboratory studies. Variation from unaccounted environmental variables may have obfuscated divergent behavioral tactics. Nevertheless, the behaviors we document here provide insights on the range of defensive behaviors exhibited by C. sculpturatus under natural environmental conditions, including the frequency of dry stings (11.8%) to the membrane-covered beakers. Full article

(This article belongs to the Section Animal Venoms)

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16 pages, 3855 KiB

Open AccessArticle

A 20-Year Retrospective Analysis of Plant Poisoning Cases at the Naval Hospital, Varna, Bulgaria

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

Toxins 2025, 17(4), 197; https://doi.org/10.3390/toxins17040197 - 12 Apr 2025

Viewed by 603

Abstract

The nature and epidemiology of plant intoxications are still not well understood, with recent data being limited. The present study aims to report cases of plant poisoning in the clinical practice of the Clinical Toxicology Department at the Naval Hospital—Varna, Bulgaria, over a [...] Read more.

The nature and epidemiology of plant intoxications are still not well understood, with recent data being limited. The present study aims to report cases of plant poisoning in the clinical practice of the Clinical Toxicology Department at the Naval Hospital—Varna, Bulgaria, over a 20-year period (2003–2023). A documentary retrospective analysis of the hospitalized cases of poisoning with poisonous plants and their grouping into toxidromes was performed. During the study period, patients with plant poisoning admitted to our hospital unit accounted for 0.35% of a total of 12,857 hospitalized individuals. The distribution across the toxidromes based on clinical presentation revealed the highest frequency of anticholinergic, cyanogen, and ricin toxidromes. The majority of the intoxications resulted from unintentional exposure to plant toxins in adult individuals. Most cases followed a mild to severe clinical course, with patient discharge occurring between 2 and 5 days. No fatalities were recorded, thanks to the reported treatment methods. A relatively low incidence of plant-related poisonings was observed, with their predominant manifestations affecting the gastrointestinal, nervous, and cardiovascular systems. Increased reporting of epidemiological data and clinical experiences in the management of plant intoxications would substantially enhance researchers’ understanding of them and facilitate the development of a standardized treatment protocol. Full article

(This article belongs to the Section Plant Toxins)

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22 pages, 4041 KiB

Open AccessArticle

Histone Methyltransferases AcDot1 and AcRmtA Are Involved in Growth Regulation, Secondary Metabolism, and Stress Response in Aspergillus carbonarius

by Angelo Agnusdei, Adrián González-García, Donato Gerin, Stefania Pollastro, Francesco Faretra, Luis González-Candelas and Ana-Rosa Ballester

Toxins 2025, 17(4), 196; https://doi.org/10.3390/toxins17040196 - 12 Apr 2025

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Abstract

Histone post-translational modifications (HPTMs) can affect gene expression by rearranging chromatin structure. Between these, histone methylation is one of the most studied in filamentous fungi, and different conserved domains coding for methyltransferase were found in Aspergillus spp. genomes. In this work, the role [...] Read more.

Histone post-translational modifications (HPTMs) can affect gene expression by rearranging chromatin structure. Between these, histone methylation is one of the most studied in filamentous fungi, and different conserved domains coding for methyltransferase were found in Aspergillus spp. genomes. In this work, the role of the histone methyltransferases AcDot1 and AcRmtA in the mycotoxigenic fungus Aspergillus carbonarius was investigated, obtaining knockout or overexpression mutants through Agrobacterium tumefaciens-mediated transformation (ATMT). A. carbonarius is responsible for grape-bunch rot, representing the major source of ochratoxin A (OTA) contamination on grapes. In vivo conditions, the deletion of Acdot1 or AcrmtA resulted in upregulation of growth when the isolates were cultivated on a minimal medium. The influence of Acdot1 on the OTA biosynthesis was differently affected by culture conditions. On rich media, an increase in OTA accumulation was observed, while on minimal medium, lower OTA concentrations were reported. The deletion of AcrmtA always resulted in lower OTA accumulation. However, the expression of OTA biosynthesis genes was regulated by both histone methyltransferases. Of the six analyzed OTA genes, three of them showed altered expression in the knockout mutants, and otaB and otaR1 were common between both mutants. Furthermore, both AcDot1 and AcRmtA play a role in oxidative stress response, induced by 1 mM hydrogen peroxide, by modulating growth, conidiation and OTA biosynthesis. Neither the deletion nor the overexpression of the Acdot1 or AcrmtA affected virulence, while both the sporulation and OTA production were negatively affected in vivo by the deletion of AcrmtA. Full article

(This article belongs to the Section Mycotoxins)

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Graphical abstract

23 pages, 935 KiB

Open AccessReview

Reviewing Evidence for Disturbance to Coral Reefs Increasing the Risk of Ciguatera

by Michael J. Holmes and Richard J. Lewis

Toxins 2025, 17(4), 195; https://doi.org/10.3390/toxins17040195 - 11 Apr 2025

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Abstract

The hypothesis that disturbance to coral reefs creates new surfaces that increase the risk of ciguatera is premised upon the increased algal substrates that develop on these surfaces being colonised by high ciguatoxin (CTX)-producing Gambierdiscus species that proliferate and enter the ciguatera food [...] Read more.

The hypothesis that disturbance to coral reefs creates new surfaces that increase the risk of ciguatera is premised upon the increased algal substrates that develop on these surfaces being colonised by high ciguatoxin (CTX)-producing Gambierdiscus species that proliferate and enter the ciguatera food chain. Current evidence indicates that new algal substrates are indeed rapidly colonised by Gambierdiscus. However, the requirement that these Gambierdiscus species include at least one that is a significant (high) CTX-producer is more likely a limiting step. While ambient environmental conditions impact the capacity of Gambierdiscus to bloom, factors that limit the growth of the bloom could influence (typically increase) the flux of CTX entering marine food chains. Additionally, new algal substrates on damaged reefs can be preferentially grazed to funnel ciguatoxins from Gambierdiscus to herbivores in disturbed reef areas. In societies consuming second trophic level species (herbivores, grazers, and detritivores), such funnelling of CTX would increase the risk of ciguatera, although such risk would be partially offset over time by growth (toxin-dilution) and depuration. Here, we review evidence for six potential mechanisms to increase ciguatera risk from disturbance to coral reefs and suggest a hypothesis where ecosystem changes could increase the flux of CTX to groupers through a shift in predation from predominately feeding on planktonic-feeding prey to mostly feeding on benthic-feeding prey, increasing the potential for CTX to accumulate. Evidence for this hypothesis is stronger for the Pacific and Indian Oceans, and it may not apply to the Caribbean Sea/Atlantic Ocean. Full article

(This article belongs to the Special Issue Special Issue in Honor of Prof. Richard J. Lewis: From Ciguatera to Conotoxin Research)

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24 pages, 1322 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

Viewed by 335

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 clam; [...] 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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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

Viewed by 1390

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

Viewed by 752

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

Viewed by 317

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

Cited by 1 | Viewed by 358

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

Viewed by 503

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

Viewed by 447

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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19 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

Viewed by 285

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

Viewed by 384

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

Viewed by 2030

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

Viewed by 400

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

Viewed by 576

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

Viewed by 986

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

Viewed by 333

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

Viewed by 539

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

Viewed by 1381

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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Toxins, Volume 17, Issue 4 (April 2025) – 59 articles

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