Toxins

17 pages, 2819 KiB

Open AccessArticle

Isolation and Mechanistic Investigation of the Efficient Zearalenone-Removing Strain Bacillus licheniformis YJ25

by Yuting Wu, Feina Wu, Pan Zhao, Yan Gao, Mengyao Li, Mengjiao Luo, Qian Zhou, Siyuan Zhou, Xinhui Li, Yaling Hong, Yang Wu, Zhaorong Zhou, Yang Liu, Yandong Xia, Lijun Zou and Jia Yin

Toxins 2025, 17(6), 263; https://doi.org/10.3390/toxins17060263 (registering DOI) - 23 May 2025

Abstract

Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin produced by Fusarium graminearum species, poses a significant threat to both human food safety and animal feed quality. In this study, we isolated a strain, designated as Bacillus licheniformis YJ25, from a contaminated moldy corn sample, demonstrating [...] Read more.

Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin produced by Fusarium graminearum species, poses a significant threat to both human food safety and animal feed quality. In this study, we isolated a strain, designated as Bacillus licheniformis YJ25, from a contaminated moldy corn sample, demonstrating substantial effectiveness in removing ZEN. Our findings revealed that YJ25’s ZEN removal occurs primarily through cell wall adsorption, with enzymatic degradation representing a potential mechanism. In practical applications, enzymatic degradation may yield metabolites with heightened toxicity. However, liquid chromatography–mass spectrometry (LC–MS) analysis revealed that ZEN was not converted into α-/β-zearalenol (α-/β-ZEL) or α-/β-zearalanol (α-/β-ZAL) by YJ25, substantiating the safety profile of YJ25 in the removal of ZEN. Our mechanistic investigations revealed that the cell wall components peptidoglycan and teichoic acid serve as the primary binding sites for ZEN adsorption. Fourier-transform infrared spectroscopy (FTIR) analysis identified O-H, C-H, C=O, and C-O as the principal functional groups participating in the cell wall adsorption process. These investigations establish a scientific foundation for the prospective application of this strain as an efficient biological detoxification agent in food and feed safety management systems. Full article

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28 pages, 5963 KiB

Open AccessArticle

Computational Modeling of Low-Abundance Proteins in Venom Gland Transcriptomes: Bothrops asper and Bothrops jararaca

by Joseph Espín-Angulo and Doris Vela

Toxins 2025, 17(6), 262; https://doi.org/10.3390/toxins17060262 - 22 May 2025

Abstract

Snake venoms contain numerous toxic proteins, but low-abundance proteins often remain uncharacterized due to identification challenges. This study employs a bioinformatics approach to identify and structurally model low-abundance proteins from the venom gland transcriptomes of Bothrops asper and Bothrops jararaca. Using tools [...] Read more.

Snake venoms contain numerous toxic proteins, but low-abundance proteins often remain uncharacterized due to identification challenges. This study employs a bioinformatics approach to identify and structurally model low-abundance proteins from the venom gland transcriptomes of Bothrops asper and Bothrops jararaca. Using tools such as tblastn, Jalview, and CHIMERA, we analyzed sequences and structural features of proteins including arylsulfatase, CRISP (Cysteine-Rich Secretory Protein), von Willebrand factor type D (vWFD), and dihydroorotate dehydrogenase (DHODH), and identified potential new isoforms of SVMP-PIIIb (Ba_1) and botrocetin in B. asper. Protein models were generated with AlphaFold2, compared with crystallized structures from the Protein Data Bank (PDB), and validated using Procheck, ERRAT, and Verify3D. Conserved motifs and domains were annotated through Pfam and InterPro, revealing structural elements that suggest possible roles in venom physiology and toxicity. These findings emphasize the potential of computational biology to characterize structurally relevant but experimentally inaccessible venom proteins, and to lay the groundwork for future functional validation. Full article

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

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22 pages, 685 KiB

Open AccessReview

Mycotoxins in Broiler Production: Impacts on Growth, Immunity, Vaccine Efficacy, and Food Safety

by Ramona Maria Olariu, Nicodim Iosif Fiţ, Cosmina Maria Bouari and George Cosmin Nadăş

Toxins 2025, 17(6), 261; https://doi.org/10.3390/toxins17060261 - 22 May 2025

Abstract

Mycotoxins are secondary fungal metabolites that frequently contaminate poultry feed, posing significant risks to animal health, productivity, and food safety. In broiler production, mycotoxins such as aflatoxins, trichothecenes, fumonisins, ochratoxin A, deoxynivalenol, and zearalenone have been shown to impair growth performance, damage key [...] Read more.

Mycotoxins are secondary fungal metabolites that frequently contaminate poultry feed, posing significant risks to animal health, productivity, and food safety. In broiler production, mycotoxins such as aflatoxins, trichothecenes, fumonisins, ochratoxin A, deoxynivalenol, and zearalenone have been shown to impair growth performance, damage key organs, and disrupt immune function. This review explores the multifaceted impact of mycotoxin exposure in broilers, with particular emphasis on immunosuppression, decreased vaccine efficacy, and increased vulnerability to infectious diseases, including coccidiosis, salmonellosis, E. coli, and viral infections like infectious bursal disease and infectious laryngotracheitis. Mycotoxin contamination in poultry feed can lead to direct economic losses through reduced feed conversion efficiency, increased mortality, and reproductive disorders, while also resulting in the transfer of toxic residues into meat and eggs, thereby threatening consumer health. The review further examines the synergistic interactions between mycotoxins and pathogens, the physiological and histopathological changes in exposed birds, and the implications for public health. Finally, it discusses current mitigation strategies, including mycotoxin binders, probiotics, and regulatory approaches to reduce exposure. An integrated management strategy combining feed hygiene, monitoring, and targeted nutritional interventions is essential to safeguard poultry health, enhance vaccine responses, and ensure the safety of poultry-derived food products. This review offers actionable insights for veterinarians, nutritionists, and policymakers, reinforcing the importance of mycotoxin mitigation strategies within a One Health framework. Full article

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

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16 pages, 4520 KiB

Open AccessArticle

StingReady: A Novel Device for Controlled Insect Sting Challenge—From Field Capture to Clinical Application

by Xesús Feás, Margarita Armisén, Sara López-Freire, Manuela Alonso-Sampedro and Carmen Vidal

Toxins 2025, 17(6), 260; https://doi.org/10.3390/toxins17060260 - 22 May 2025

Abstract

Reliable assessment of protection in venom immunotherapy (VIT) patients remains a clinical challenge, especially due to the limitations of conventional sting challenge tests (SCTs), which require complex insect handling and may compromise test accuracy. This study introduces StingReady, a novel, user-friendly device designed [...] Read more.

Reliable assessment of protection in venom immunotherapy (VIT) patients remains a clinical challenge, especially due to the limitations of conventional sting challenge tests (SCTs), which require complex insect handling and may compromise test accuracy. This study introduces StingReady, a novel, user-friendly device designed to streamline the SCT process by enabling safe, efficient, and minimally manipulative exposure to hymenopteran stings. For the first time, StingReady was applied to conduct SCTs with Vespa velutina, an invasive hornet species of increasing clinical relevance. The device was tested in a real-world setting at Belvís Park in Santiago de Compostela, Spain, where hornets were successfully captured and transported to the hospital without anesthesia or limb removal. The design features adjustable mesh sizes, allowing compatibility with various hymenopteran taxa. Using StingReady, nine patients underwent SCTs with no need for direct insect handling during the hospital procedure. The process improved patient safety and comfort while preserving the insect’s natural stinging behavior, thereby enhancing test reliability. This study demonstrates that StingReady significantly improves SCT methodology, offering a practical, reproducible, and ethically sound alternative for evaluating VIT efficacy across diverse hymenopteran species. Full article

(This article belongs to the Section Animal Venoms)

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18 pages, 3255 KiB

Open AccessArticle

Growth Response and Cell Permeability of the Fish-Killing Phytoflagellate Heterosigma akashiwo Under Projected Climate Conditions

by Malihe Mehdizadeh Allaf and Charles G. Trick

Toxins 2025, 17(5), 259; https://doi.org/10.3390/toxins17050259 - 21 May 2025

Abstract

Climate change and anthropogenic alterations in biogeochemical cycles are intensifying the frequency, duration, and potential toxicity of harmful algal blooms (HABs) in marine ecosystems. However, these effects are highly variable and depend on species identity, strain-specific traits, and local environmental conditions. Key drivers [...] Read more.

Climate change and anthropogenic alterations in biogeochemical cycles are intensifying the frequency, duration, and potential toxicity of harmful algal blooms (HABs) in marine ecosystems. However, these effects are highly variable and depend on species identity, strain-specific traits, and local environmental conditions. Key drivers include rising sea surface temperatures, changes in salinity resulting from altered precipitation patterns and runoff, and elevated CO₂ levels leading to ocean acidification. Heterosigma akashiwo, a euryhaline raphidophyte responsible for the widespread killing of fish, is particularly responsive to these changes. This study investigated the combined effects of temperature, salinity, and CO₂ concentration on the growth, yield, and cell membrane permeability of H. akashiwo using a Design of Experiment (DOE) approach. DOE facilitates a detailed and systematic analysis of multifactorial interactions, enabling a deeper understanding of complex relationships while maximizing efficiency and minimizing the use of experimental resources. The results revealed that growth and yield were maximized at higher temperatures and salinities, whereas cell permeability increased under cooler, less saline, and lower CO₂ conditions. These findings suggest that projected future ocean conditions may enhance biomass production while potentially reducing cellular permeability and, by extension, toxicity. This study highlights the value of the DOE framework in identifying key interactions among environmental drivers of HABs, offering a practical foundation for future predictive modeling under climate change scenarios. Full article

(This article belongs to the Special Issue Mechanisms Underlying Metabolic Regulation by Marine Toxins)

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13 pages, 1599 KiB

Open AccessReview

Botulinum Toxin in Pain-Related Post-Stroke Limb Spasticity: A Meta-Analysis of Early and Late Injections

by Frances Marie Tamayo, Raymond Rosales, Jörg Wissel, Bo Biering-Sørensen, Joshua Nathaniel Ellano and David Simpson

Toxins 2025, 17(5), 258; https://doi.org/10.3390/toxins17050258 - 21 May 2025

Abstract

Spasticity is a common complication associated with stroke, and around 72% of stroke patients will develop pain during the disease. Botulinum toxin (BoNT) is a safe and efficacious treatment for spasticity and can improve associated complications, including pain. Hence, this meta-analysis aims to [...] Read more.

Spasticity is a common complication associated with stroke, and around 72% of stroke patients will develop pain during the disease. Botulinum toxin (BoNT) is a safe and efficacious treatment for spasticity and can improve associated complications, including pain. Hence, this meta-analysis aims to establish whether BoNT can reduce pain-related post-stroke spasticity (pPSS) in the early treatment period (<12 weeks post-stroke) or in the late period (>12 weeks post-stroke) based on the available evidence. This study also aims to establish the dose–response relationship of BoNT-A in pPSS. Based on pooled data from multiple studies, there is no significant difference in the scores measuring pPSS between patients who received early BoNT-A injections and those who received a placebo. This finding suggests that within the early treatment period, BoNT-A may not be more effective than a placebo in reducing pPSS. However, it is important to note that the data for early BoNT-A injections are limited, indicating that research is needed to draw definitive conclusions [z = 3.90 (p < 0.0001)]. While BoNT-A appears somewhat more effective than a placebo in the late phase, as indicated by the small to moderate positive z value, there is not enough evidence to confidently claim superiority over a placebo [z = 1.48 (p = 0.14)]. Full article

(This article belongs to the Special Issue Botulinum Toxins: New Uses in the Treatment of Diseases (2nd Edition))

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16 pages, 1260 KiB

Open AccessReview

Clinical Evidence of Bee Venom Acupuncture for Ankle Pain: A Review of Clinical Research

by Soo-Hyun Sung, Hyein Jeong, Jong-Hyun Park, Minjung Park and Gihyun Lee

Toxins 2025, 17(5), 257; https://doi.org/10.3390/toxins17050257 - 21 May 2025

Abstract

The prevalence of ankle pain in adults is 9–15%, with up to 45% of sports-related injuries attributed to ankle pain and injuries. If ankle pain is not controlled in a timely manner, it can lead to ankle instability, resulting in further damage, recurrence [...] Read more.

The prevalence of ankle pain in adults is 9–15%, with up to 45% of sports-related injuries attributed to ankle pain and injuries. If ankle pain is not controlled in a timely manner, it can lead to ankle instability, resulting in further damage, recurrence of pain, and secondary injuries. The present study aimed to assess the therapeutic potential and safety profile of bee venom acupuncture (BVA) in the management of ankle pain. Ten electronic databases were searched for articles published up to March 2025. We included clinical studies that utilized BVA for the treatment of ankle pain and studies that included pain- and function-related assessment tools. The safety of bee venom acupuncture (BVA) was assessed by extracting adverse events from the included studies and categorizing them according to the Common Terminology Criteria for Adverse Events (CTCAE). A total of 14 clinical studies were selected, of which 9 were case reports, 2 were case-controlled clinical trials (CCTs), and 3 were randomized controlled trials (RCTs). The conditions causing ankle pain were mostly traumatic (42.9%), followed by inflammatory (21.4%) and neuropathic disorders (14.3%). BVA was applied at concentrations ranging from 0.05 to 0.5 mg/mL, with a per-session volume ranging from 0.04 to 2.5 mL. In most studies, BVA was reported to improve both ankle pain and function simultaneously. Among the 14 studies, four participants reported adverse events following BVA treatment, all of which were classified as grade 1 or grade 2, indicating mild to moderate severity. This review suggests that BVA may be recommended for controlling ankle pain based on clinical evidence. However, the number of high-quality RCTs is limited, and half of the studies did not report side effects, indicating the need for further clinical research to verify its safety and efficacy. Full article

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

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25 pages, 1538 KiB

Open AccessSystematic Review

Effect of Ochratoxin A (OTA) on the Immune System: A Systematic Review

by Yusif Mubarik, Shadrach Tetteh Boyetey, Anastasia Rosebud Aikins and Mohamed Mutocheluh

Toxins 2025, 17(5), 256; https://doi.org/10.3390/toxins17050256 - 20 May 2025

Abstract

Ochratoxin A (OTA) is a mycotoxin with different adverse health effects. The authors conducted a systematic review to evaluate the effects of OTA on the immune system, with more emphasis on its effects on immune system organs, innate and adaptive immunity and related [...] Read more.

Ochratoxin A (OTA) is a mycotoxin with different adverse health effects. The authors conducted a systematic review to evaluate the effects of OTA on the immune system, with more emphasis on its effects on immune system organs, innate and adaptive immunity and related signaling pathways. Studies have demonstrated that exposure to OTA disrupts the functions of immune system organs, resulting in weight loss, histological lesions and a decrease in antibody-secreting cells. There is evidence that OTA impairs epithelial barrier integrity and macrophage function and induces elevated secretion of pro-inflammatory cytokines. In adaptive immunity, OTA regulates T-cell differentiation, particularly Th1 and Th17 subsets, and adversely impacts humoral immunity, ultimately leading to immune suppression. Full article

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

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25 pages, 723 KiB

Open AccessSystematic Review

Systematic Review on CyanoHABs in Central Asia and Post-Soviet Countries (2010–2024)

by Kakima Kastuganova, Galina Nugumanova and Natasha S. Barteneva

Toxins 2025, 17(5), 255; https://doi.org/10.3390/toxins17050255 - 20 May 2025

Abstract

Cyanobacterial harmful blooms (CyanoHABs) in lakes, estuaries, and freshwater reser-voirs represent a significant risk to water authorities worldwide due to their cyanotoxins and economic impacts. The duration, spread, and severity of CyanoHABs have markedly increased over the past decades. The article addresses CyanoHABs, [...] Read more.

Cyanobacterial harmful blooms (CyanoHABs) in lakes, estuaries, and freshwater reser-voirs represent a significant risk to water authorities worldwide due to their cyanotoxins and economic impacts. The duration, spread, and severity of CyanoHABs have markedly increased over the past decades. The article addresses CyanoHABs, cyanotoxins, and monitoring methodologies in post-Soviet and Central Asian countries. This particular re-gion was selected for the systematic review due to its relative lack of representation in global CyanoHABs reporting, particularly in Central Asia. The main aim of this systematic review was to analyze the primary literature available from 2010–2024 to examine the current situation of CyanoHAB detection, monitoring, and management in Central Asia and post-Soviet countries. Following a detailed database search in several selected data-bases (Google Scholar, Pubmed, Web of Science (WOS), Scopus, Elibrary, ENU, and KazNU) along with additional hand searching and citation searching, 121 primary arti-cles reporting 214 local cyanobacterial bloom cases were selected for this review. Aquatic cyanotoxins were reported in water bodies of eight countries, including high concentra-tions of microcystins that often exceeded reference values established by the World Health Organization (WHO). Advancing monitoring efforts in Baltic countries, Belarus, and the Russian Federation differed from only a few Central Asian reports. However, Central Asian aquatic ecosystems are especially threatened by rising anthropogenic pressures (i.e., water use, intensive agriculture, and pollution), climate change, and the lack of adequate ecological surveillance. We hypothesize that recent Caspian seal mass mortality events have been caused by a combination of infection (viral or bacterial) and exposure to algal neurotoxins resulting from harmful algal blooms of Pseudo-nitzschia. We conclude that there is an urgent need to improve the assessment of cyanobacterial blooms in Central Asia and post-Soviet countries. Full article

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

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30 pages, 5676 KiB

Open AccessReview

Insecticidal Activities of Diterpene Alkaloids in Plants of the Genera Aconitum and Delphinium

by Jinqiu Wang, Luchuan Zheng, Wenxi Huang, Linxuan Li, Jialian Yuan and Lin Chen

Toxins 2025, 17(5), 254; https://doi.org/10.3390/toxins17050254 - 20 May 2025

Abstract

As the global population grows, food security and agricultural productivity face challenges, and insect pests cause significant losses to crops. The effectiveness of traditional chemical pesticides is declining, and eco-friendly pesticides need to be developed. Diterpenoid alkaloids (DAs), natural products of plant origin, [...] Read more.

As the global population grows, food security and agricultural productivity face challenges, and insect pests cause significant losses to crops. The effectiveness of traditional chemical pesticides is declining, and eco-friendly pesticides need to be developed. Diterpenoid alkaloids (DAs), natural products of plant origin, have attracted attention due to their low environmental risks. Here we review the classification, structure, insecticidal and anti-feeding activities of diterpenoid alkaloids, as well as the current state of research on these chemicals. Studies have shown that C₁₉- and C₂₀-diterpenoid alkaloids show significant activity against a variety of insects, but there are still limited studies on C₁₈-diterpenoid alkaloids. Therefore, through in-depth research on diterpenoid alkaloids, we have discovered that there are various compounds with high efficiency and specificity in insecticidal and antifeedant activities among C₁₉- and C₂₀-diterpenoid alkaloids, which exhibit high selectivity and efficiency towards target pests. This paper emphasizes the potential of diterpenoid alkaloids as novel biopesticides and highlights the need to combine new technologies to conduct further systematic evaluation and screening of these compounds. This work provides new ideas for the development of environmentally friendly pesticides and contributes to sustainable agricultural practices. Full article

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17 pages, 3704 KiB

Open AccessArticle

Heterologous Overexpression of Cytochrome P450BM3 from Bacillus megaterium and Its Role in Gossypol Reduction

by Wenpeng Fan, Jingjing Cui, Tongxiang Xu, Shiheng Xu, Zulibina Ainiwaer, Qiyu Luo and Caidie Wang

Toxins 2025, 17(5), 253; https://doi.org/10.3390/toxins17050253 - 20 May 2025

Abstract

Gossypol is a polyphenolic toxic compound present in cotton plants. To determine whether the candidate cytochrome P450BM3 enzymes could reduce gossypol in vitro, functional recombinant cytochrome P450BM3 enzymes were successfully expressed in E. coli. Site-directed mutagenesis generated mutants (R162H, R162K, Q129H, Q129N) [...] Read more.

Gossypol is a polyphenolic toxic compound present in cotton plants. To determine whether the candidate cytochrome P450BM3 enzymes could reduce gossypol in vitro, functional recombinant cytochrome P450BM3 enzymes were successfully expressed in E. coli. Site-directed mutagenesis generated mutants (R162H, R162K, Q129H, Q129N) to explore structural determinants of catalytic efficiency. Both wild-type P450BM3 and mutants exhibited significant ability to reduce gossypol levels, with R162H and R162K showing 33.4% and 24.2% reduced catalytic efficiency compared with the wild-type enzyme, respectively. Q129H and Q129N mutants maintained comparable catalytic efficiency to the wild type. Metabolomic profiling revealed two distinct reducing pathways catalyzed by wild-type P450BM3 and its mutants (R162H/Q129H), involving decarboxylation, hydroxylation, and C-C bond cleavage. This study demonstrated the feasibility of P450BM3 as a highly efficient biocatalyst for reducing gossypol levels, speculated that Arg162 might be a critical active residue, and hypothesized the potential pathways by which P450BM3 catalyzes the reduction of gossypol content, thereby providing a theoretical foundation for the enzymatic reduction of gossypol. Full article

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13 pages, 1836 KiB

Open AccessArticle

A Comprehensive Probabilistic Risk Assessment Strategy of Aflatoxin B1 Exposure from Medical Areca Nuts Consumption

by Tian-Tian Zuo, Ya-Zhong Zhang, Hong-Yan Zhai, Rong-Yao Chen, Jing-Zhe Pu, Hong-Yu Jin, Jing Liu, Xian-Long Cheng and Feng Wei

Toxins 2025, 17(5), 252; https://doi.org/10.3390/toxins17050252 - 19 May 2025

Abstract

Aflatoxin B₁ (AFB₁) is widely found and substantially impends public health. The current work aimed to assess AFB₁ in medical areca nuts in China. The average content of AFB₁ was 13.0 μg/kg, and the maximum content was 146.0 [...] Read more.

Aflatoxin B₁ (AFB₁) is widely found and substantially impends public health. The current work aimed to assess AFB₁ in medical areca nuts in China. The average content of AFB₁ was 13.0 μg/kg, and the maximum content was 146.0 μg/kg. Furthermore, a comprehensive probabilistic risk assessment approach considering combined utilization of a Monte Carlo simulation with the margin of exposure (MOE) and quantitative liver cancer risk (HCC) strategies was developed for assessing the human health risk of AFB₁ from consuming medical areca nuts for the first time. The MOE values of AFB₁ in samples for no more than the 75th percentile were less than the threshold of 10,000 for both men and women. The estimated 90th percentile to the maximum of the HCC values for males and the estimated 75th percentile to the maximum of the HCC values for females were higher than one in a million upon exposure to medical areca nuts, indicating an unacceptable liver cancer risk. Sensitivity analysis demonstrated that for both MOE and HCC approaches, AFB₁ content was the parameter with the greatest effects on the results, followed by the exposure frequency (EF) and daily intake rate (IR). This study is the first of this kind, demonstrating the applicability of stochastic exposure evaluation techniques for the precise and scientific assessment of the health risk of AFB₁ in medical areca nuts, with the main purpose of minimizing human cancer risk. Full article

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23 pages, 3679 KiB

Open AccessArticle

Impact of Low-Level Ergot Alkaloids and Endophyte Presence in Tall Fescue Grass on the Metabolome and Microbiome of Fall-Grazing Steers

by Ignacio M. Llada, Jeferson M. Lourenco, Madison M. Dycus, Jessica M. Carpenter, Zachery R. Jarrell, Dean P. Jones, Garret Suen, Nicholas S. Hill and Nikolay M. Filipov

Toxins 2025, 17(5), 251; https://doi.org/10.3390/toxins17050251 - 17 May 2025

Abstract

Fescue toxicosis (FT) is a mycotoxin-related disease caused by the ingestion of tall fescue, naturally infected with the ergot alkaloid (EA)-producing endophyte Epichloë coenophiala. Some grazing on endophyte-free (E−) or non-toxic (NT), commercial endophyte-infected pastures takes place in the US as well. [...] Read more.

Fescue toxicosis (FT) is a mycotoxin-related disease caused by the ingestion of tall fescue, naturally infected with the ergot alkaloid (EA)-producing endophyte Epichloë coenophiala. Some grazing on endophyte-free (E−) or non-toxic (NT), commercial endophyte-infected pastures takes place in the US as well. Earlier, we found that grazing on toxic fescue with low levels of EAs during fall affects thermoregulation, behavior, and weight gain. Building on these findings, the current study aimed to investigate how the presence of low EA-producing E+ or NT endophytes can influence animal metabolome, microbiome, and, ultimately, overall animal health. Eighteen Angus steers were placed on NT, E+, and E− fescue pastures for 28 days. Urine, rumen fluid (RF), rumen solid (RS), and feces were collected pre-exposure, and on days 2, 7, 14, 21, and 28. An untargeted high-resolution metabolomics approach was used to analyze urine and RF, while 16S rRNA-based next-generation sequencing (NGS) was used to examine RF, RS, feces, and fescue plant microbiomes. While alpha- or beta-microbiota diversity across all analyzed matrices were unaffected, there were specific effects of E+ on the relative abundance of some taxa (i.e., Prevotellaceae). Additionally, E+ grazing impacted aromatic amino acid metabolism in the urine and the metabolism of lipids in both the RF and urine. In both matrices, trace amine-related metabolic features differed markedly between E+ and the other groups. Compared to the endophyte-free group, endophyte presence, whether novel or toxic, influenced amino acid and carbohydrate metabolism, as well as unsaturated fatty acid biosynthesis. These findings suggest that low-EA-producing and non-toxic endophytes in fescue have more prominent effects on the metabolome than the microbiome, and this metabolome perturbation might be associated with decreased performance and reported physiological signs of FT. Full article

(This article belongs to the Section Mycotoxins)

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15 pages, 2172 KiB

Open AccessArticle

Ustilaginoidin D Induces Acute Toxicity and Hepatotoxicity in Mice

by Bo Wang, Xiaolong Bai, Min Zhang, Xiangxiang Liu, Muhammad Zulqar Nain Dara, Lingjing Liu, Mingming Ou, Dayong Li, Jiyang Wang, Ling Liu and Wenxian Sun

Toxins 2025, 17(5), 250; https://doi.org/10.3390/toxins17050250 - 17 May 2025

Abstract

Ustilaginoidin D is a type of bis-naphtho-γ-pyrone mycotoxin produced by Ustilaginoidea virens, the causal agent of rice false smut. Although previous studies have demonstrated the inhibitory effect of ustilaginoidin D on ATP synthesis and cancer cell growth in mice, its specific health risks [...] Read more.

Ustilaginoidin D is a type of bis-naphtho-γ-pyrone mycotoxin produced by Ustilaginoidea virens, the causal agent of rice false smut. Although previous studies have demonstrated the inhibitory effect of ustilaginoidin D on ATP synthesis and cancer cell growth in mice, its specific health risks remain unclear. Here, we reveal that ustilaginoidin D is highly toxic to mice with an LD₅₀ value of 213 mg /kg·bw. Dose-dependent weight loss and liver damage were observed, accompanied by altered markers of liver cell damage, including the enzyme activities of alanine aminotransferase and aspartate aminotransferase and the content of glutathione in mouse liver. RNA-seq analysis of liver tissues from mice treated with 150 mg of ustilaginoidin D/kg·bw identified significant changes in gene expression profiles, with differentially expressed genes enriched in cancer-related pathways, hypertrophic cardiomyopathy, and metabolic pathways. RT-qPCR data are highly consistent with transcriptome analysis in expression profiles of 22 chemical-carcinogenesis-associated genes. These findings indicate that ustilaginoidin D induces acute toxicity and liver dysfunction in mice, raising serious concerns about its threat to human health. Full article

(This article belongs to the Special Issue Occurrence, Toxicity, Metabolism, Analysis and Control of Mycotoxins)

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17 pages, 2725 KiB

Open AccessReview

Botulinum Toxin for Bruxism: An Overview

by Manuella Salm Coelho, Júlia Meller Dias de Oliveira, Helena Polmann, Patrícia Pauletto, Cristine Miron Stefani, Lara Catarine De Luca Maciel and Graziela De Luca Canto

Toxins 2025, 17(5), 249; https://doi.org/10.3390/toxins17050249 - 16 May 2025

Abstract

This overview aimed at assessing botulinum toxin type A (BoNT-A)’s effectiveness in managing bruxism compared to a placebo, the absence of treatment, or other interventions in adults. Only systematic reviews (SRs), with or without a meta-analysis, were included. A comprehensive literature search was [...] Read more.

This overview aimed at assessing botulinum toxin type A (BoNT-A)’s effectiveness in managing bruxism compared to a placebo, the absence of treatment, or other interventions in adults. Only systematic reviews (SRs), with or without a meta-analysis, were included. A comprehensive literature search was conducted on 16 June 2024, encompassing seven databases and grey literature sources. Experts and reference lists of the included SRs were also consulted. Study selection was conducted in two phases by two independent authors. Methodological quality was evaluated using AMSTAR-2. Overlap was assessed using the corrected covered area. The 14 included SRs addressed several outcomes. In most studies, BoNT-A showed effectiveness in reducing pain (n = 10), the frequency of bruxism events (n = 7), and the maximum bite force (n = 5). None of the SRs effectively evaluated BoNT-A’s impact on functional movements. All of the included SRs scored “critically low” and “low” confidence rates in AMSTAR-2. Only one SR attempted to assess the certainty of the evidence, although unsuccessfully. The overlap across included SRs was rated as “high”, with a corrected covered area of 10.11%. The current systematic reviews on BoNT-A for bruxism lack methodological quality, limiting the reliability of their findings. Despite evidence indicating a potential reduction in pain, bruxism events, and the maximum bite force, methodological limitations prevent definitive conclusions from being drawn. High-quality research with standardized methodologies is essential to establish BoNT-A’s efficacy and support evidence-based clinical practice. Full article

(This article belongs to the Section Bacterial Toxins)

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19 pages, 3329 KiB

Open AccessArticle

Response Surface Methodology Optimization of Time-Resolved Fluorescence Immunoassay for Rapid Detection of AflatoxinB₁ in Yellow Rice Wine

by Mengjie Zhu, Dun Wang, Du Wang, Jing Dong, Xue Wang, Qi Zhang and Man Xiao

Toxins 2025, 17(5), 248; https://doi.org/10.3390/toxins17050248 - 16 May 2025

Abstract

Yellow rice wine is susceptible to aflatoxinB₁ (AFB₁) contamination, yet existing detection technologies suffer from limitations such as high false-positive rates, cumbersome operational protocols, or elevated costs, rendering them inadequate for large-scale screening requirements. Consequently, the development of a highly [...] Read more.

Yellow rice wine is susceptible to aflatoxinB₁ (AFB₁) contamination, yet existing detection technologies suffer from limitations such as high false-positive rates, cumbersome operational protocols, or elevated costs, rendering them inadequate for large-scale screening requirements. Consequently, the development of a highly sensitive and rapid detection method for AFB₁ is urgently needed to provide technical support for quality supervision and risk assessment of yellow rice wine. In this study, AFB₁ detection was performed using time-resolved fluorescence immunoassay technology, with quantitative analysis based on the ratio of the T signal value of the detection line to the C signal value of the quality control line and the natural logarithmic value of the standard solution concentration. Statistical experimental designs were used to optimize the process of this rapid detection of AFB₁ in yellow rice wine. The most important factors influencing recovery rate (p < 0.05), as identified by a two-level Plackett-Burman design with 11 variables, were methanol-water volume fraction, sample to extraction solvent ratio, heating temperature, and heating time. The steepest ascent method was employed to identify the optimal regions for these four key factors. Central composite design (CCD) coupled with response surface methodology (RSM) was subsequently utilized to further explore the interactive effects among variables and determine their optimal values that maximize the recovery rate. The analysis results indicated that interactions between methanol-water volume fraction and other three factors–sample to extraction solvent ratio, heating temperature, heating time–affected the response variable (recovery rate) significantly. The predicted results showed that the maximum recovery rate of AFB₁ (97.35%) could be obtained under the optimum conditions of a methanol-water volume fraction of 78%, a sample to extraction solvent ratio of 1:3.2, a heating temperature of 34 °C, and a heating time of 6.4 min. These predicted values were further verified by validation experiments. The excellent correlation between predicted and experimental values confirmed the validity and practicability of this statistical optimum strategy. Optimal conditions obtained in this experiment laid a good foundation for further use of time-resolved fluorescence immunoassay for rapid detection of AFB₁ in yellow rice wine, demonstrating broad application prospects. Full article

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12 pages, 1959 KiB

Open AccessArticle

Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection

by Yi-Zhe Zhang, Yi Yao, Kai-Ping Zhang, Jia-Qi Liang, Jia-Ju Zhong, Zhong-Feng Li, Hai-Jiao Li and Fei Xu

Toxins 2025, 17(5), 247; https://doi.org/10.3390/toxins17050247 - 16 May 2025

Abstract

Amanita species are widely distributed worldwide. Many of these species are poisonous and can cause health problems, resulting in morbidity and mortality. The toxins responsible for poisoning are amatoxins, aminohexadienoic acid, ibotenic acid, muscimol and muscarines, which damage the liver, kidney, central nervous [...] Read more.

Amanita species are widely distributed worldwide. Many of these species are poisonous and can cause health problems, resulting in morbidity and mortality. The toxins responsible for poisoning are amatoxins, aminohexadienoic acid, ibotenic acid, muscimol and muscarines, which damage the liver, kidney, central nervous system and parasympathetic nervous system. In recent years, several toxins have been discovered from different poisonous mushrooms. In this study, multiwalled carbon nanotube purification and ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used for the sensitive detection and targeted quantitative screening of 12 mushroom toxins (muscarine, two isoxazole derivatives, three tryptamine alkaloids, three amatoxins and three phallotoxins) from Amanita citrina, A. citrina var. grisea and A. sinocitrina. This study found that buiotenine, one of the tryptamine alkaloids, was detected in A. citrina and A. sinocitrina with an average content of 2.90 and 1.19–6.70 g/kg (n = 3) in the dried mushrooms, respectively. None of the 12 common toxins were discovered in A. citrina var. grisea. These results provide reference data for future research on the role of toxins in the evolution of Amanita mushrooms. Future studies should explore the biosynthetic pathways and ecological roles of these toxins in Amanita species. Full article

(This article belongs to the Special Issue Advances in Poisonous Mushrooms and Their Toxins)

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2 pages, 477 KiB

Open AccessCorrection

Correction: Di Paola et al. Impact of Mycotoxin Contaminations on Aquatic Organisms: Toxic Effect of Aflatoxin B1 and Fumonisin B1 Mixture. Toxins 2022, 14, 518

by Davide Di Paola, Carmelo Iaria, Fabiano Capparucci, Alessia Arangia, Rosalia Crupi, Salvatore Cuzzocrea, Nunziacarla Spanò, Enrico Gugliandolo and Alessio Filippo Peritore

Toxins 2025, 17(5), 246; https://doi.org/10.3390/toxins17050246 - 15 May 2025

Abstract

In the original publication [...] Full article

(This article belongs to the Section Mycotoxins)

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28 pages, 8296 KiB

Open AccessArticle

Survey of Microcystin-Producing Cyanobacteria in French Lakes of Various Trophic Status Using Environmental and Cyanobacterial Parameters and an Active Mussel Biomonitoring

by Emilie Lance, Alexandra Lepoutre, Luc Brient, Nicolas Maurin, Emmanuel Guillon, Alain Geffard and Dominique Amon-Moreau

Toxins 2025, 17(5), 245; https://doi.org/10.3390/toxins17050245 - 15 May 2025

Abstract

Microcystins (MCs), hepatotoxins produced by cyanobacteria, represent a potential threat to aquatic ecosystems and human health. Measuring various environmental and cyanobacterial parameters in water samples can be useful for monitoring water quality and assessing risk but remains a short-term approach. Beyond local risk [...] Read more.

Microcystins (MCs), hepatotoxins produced by cyanobacteria, represent a potential threat to aquatic ecosystems and human health. Measuring various environmental and cyanobacterial parameters in water samples can be useful for monitoring water quality and assessing risk but remains a short-term approach. Beyond local risk assessments, estimating global and medium-term levels of freshwater contamination by MC-producing cyanobacteria is challenging in large lakes due to the spatio-temporal variability of their proliferation and the need to multiply sampling dates and locations. In such conditions, a sentinel organism can be valuable for monitoring MCs in situ and providing a time-integrated picture of contamination levels at various stations. We previously assessed the ability of the freshwater bivalves Anodonta anatina and Dreissena polymorpha to act as biointegrators of MCs, even under low exposure levels to cyanobacteria. In this study, through a two-season investigation in several French lakes experiencing moderate cyanobacterial blooms, we evaluated the relevance of various parameters (cyanobacterial density and biovolume, chlorophyll-a, and phycocyanin) as well as the use of bivalves as indicators of medium-term freshwater contamination by MC-producing cyanobacteria. MC concentrations in cyanobacterial biomass (intracellular MCs) and in bivalves (free MCs, being unbound, and total free and protein-bound accumulated MCs) were measured alongside the characterization of phytoplankton communities. Both mussels integrated and highlighted the presence of intracellular MCs in the environment over the period between two successive water samplings, even at low contamination levels, demonstrating their suitability for in situ biomonitoring of MC-producing cyanobacteria. The results are discussed in terms of the strengths and limitations of different parameters for assessing MC contamination levels in waters depending on the objective (managing, preventing, or global evaluation) and the monitoring strategies used. Full article

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

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