Toxins

15 pages, 1482 KiB

Open AccessArticle

Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures

by Anna Maria Engel, Ahmed H. El-Khatib, Martin Bachmann, Monika Wensch-Dorendorf, Fenja Klevenhusen, Stefan Weigel, Robert Pieper and Annette Zeyner

Toxins 2025, 17(2), 46; https://doi.org/10.3390/toxins17020046 - 21 Jan 2025

Abstract

The transformation of hypoglycin A (HGA), hypoglycin B (HGB), and methylene cyclopropyl glycine (MCPrG) in ruminal fluid batch cultures was investigated, and the effect of these toxins on the batch culture microorganisms using microbial metabolites was measured. An experiment was conducted using ovine [...] Read more.

The transformation of hypoglycin A (HGA), hypoglycin B (HGB), and methylene cyclopropyl glycine (MCPrG) in ruminal fluid batch cultures was investigated, and the effect of these toxins on the batch culture microorganisms using microbial metabolites was measured. An experiment was conducted using ovine ruminal fluid batch cultures and the ANKOM RF Gas Production System over four runs, each with an incubation period of 48 h. The fermenters contained 200 mg of (i) a substrate mixture (80% cellulose, 20% starch; CSM), (ii) CSM and 1.5 mL of a solution of pure toxins (a mixture of 500 ng/mL HGA and MCPrG each; PCM), or (iii) CSM and 100 mg sycamore maple seeds (SMS). Each fermenter contained 30 mL of inoculum (ruminal fluid and buffer, 1:2 v/v). For control, autoclaved ruminal fluid was incubated with CSM, PCM, and SMS, respectively. Samples were taken from the liquid phase of the fermenters and analyzed using liquid chromatography–tandem mass spectrometry (LC/MS-MS) for sycamore maple toxins and metabolites. Microbial activity was assessed using gas production, short chain fatty acids, and NH₃ concentration. Additionally, pH and redox potentials were measured. In PCM, HGA and MCPrG concentrations rapidly decreased (p < 0.05), and were not measurable anymore after a 24 h incubation period. In SMS, the initial concentrations were 4.7 ± 1.4 µg/mL HGA, 19.9 ± 5.41 µg/mL HGB, and 1.2 ± 0.33 µg/mL MCPrG. In SMS, HGA increased in 24 h, coincidently to a decrease in HGB concentration (p < 0.05). We modeled a rapid conversion of HGB to HGA, accompanied by progressive HGA transformation. The concentration of MCPrG was constant until 4 h and decreased afterwards (p < 0.05). In SMS incubations, HGA and MCPrG concentrations of 5.6 ± 1.5 and 0.32 ± 0.090 µg/mL remained after 48 h, respectively. The HGB to HGA conversion and transformation of HGA and MCPrG also occurred in autoclaved ruminal fluid. Gas production and microbial metabolite concentrations were higher in SMS compared to CSM and PCM (p < 0.05), as the seeds were used as an additional substrate by the batch culture microorganisms. Full article

(This article belongs to the Section Plant Toxins)

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26 pages, 1815 KiB

Open AccessArticle

The First Large Identification of 3ANX and NX Producing Isolates of Fusarium graminearum in Manitoba, Western Canada

by Maria Antonia Henriquez, Srinivas Sura, Sean Walkowiak, David Kaminski, Anne Kirk, Mark W. Sumarah, Parthasarathy Santhanam, Nina Kepeshchuk, Jules Carlson, E. RoTimi Ojo, Pam de Rocquigny and Holly Derksen

Toxins 2025, 17(1), 45; https://doi.org/10.3390/toxins17010045 - 17 Jan 2025

Abstract

Fusarium head blight, caused by Fusarium graminearum, continues to be one of the most important and devastating fungal diseases on cereal grains including wheat, barley, and oat crops. F. graminearum produces toxic secondary metabolites that include trichothecene type A and type B [...] Read more.

Fusarium head blight, caused by Fusarium graminearum, continues to be one of the most important and devastating fungal diseases on cereal grains including wheat, barley, and oat crops. F. graminearum produces toxic secondary metabolites that include trichothecene type A and type B mycotoxins. There are many variants of these toxins that are produced, and in the early 2010s, a novel type A trichothecene mycotoxin known as 3ANX (7-α hydroxy,15-deacetylcalonectrin) and its deacetylated product NX (7-α hydroxy, 3,15-dideacetylcalonectrin) were identified in Minnesota, USA. In the current study, a total of 31,500 wheat spikes over a period of 6 years (2015–2020) within Manitoba, Canada, were screened for the F. graminearum pathogen, which accounted for 72.8% (2015), 98.3% (2016), 71.9% (2017), 74.4% (2018), 92.6% (2019), and 66.1% (2020) of isolations. A total of 303 F. graminearum isolates, confirmed through sequencing of the ribosomal intergenic spacer, were further investigated for variation in the gene Tri1, which was previously associated with the production of the NX toxin, as well as the accumulation of mycotoxins. A subset of these isolates, consisting of 73 isolates, which tested positive or negative for the NX-Tri1-F/R assay in this study, were cultured in vitro using rice media. Mycotoxins were quantified in these samples using mass spectrometry. Using the same rice culture, genomic DNA was isolated, and the Tri1 coding sequence along with its flanking regions (upstream and downstream of the Tri1 gene) was amplified and sequenced. Deoxynivalenol (DON) accumulated in 96% of the cultures from these isolates, while 3-acetyl deoxynivalenol (3ADON) and 3ANX mycotoxins accumulated in 66% and 63%, respectively. Nivalenol, 15-acetyl deoxynivalenol, and NX mycotoxins were detected in 62%, 36%, and 19% of samples, respectively. A significant correlation was observed between 3ADON and 3ANX (r² = 0.87), as well as between DON and 3ANX (r² = 0.89). This study highlights the first large identification of 3ANX- and NX-producing isolates of F. graminearum in Western Canada. In addition, it is the first identification of 15ADON chemotypes producing 3ANX in Western Canada and the first identification of 3ANX and NX-producing isolates in Manitoba, collected from wheat samples. Full article

(This article belongs to the Section Mycotoxins)

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19 pages, 13103 KiB

Open AccessArticle

Updated Nematocyst Types in Tentacle of Venomous Box Jellyfish, Chironex indrasaksajiae _{(Sucharitakul, 2017)} and Chiropsoides buitendijki _{(Horst, 1907)} (Cnidaria, Cubozoa) in Thai Waters

by Thippawan Yasanga, Klintean Wunnapuk, Rochana Phuackchantuck, Lakkana Thaikruea, Thunyaporn Achalawitkun, Purinat Rungraung and Sineenart Santidherakul

Toxins 2025, 17(1), 44; https://doi.org/10.3390/toxins17010044 - 17 Jan 2025

Abstract

The multiple-tentacle box jellyfish, Chironex indrasaksajiae (Sucharitakul, 2017) and Chiropsoides buitendijki (Horst, 1907), are venomous species found in Thai waters. They are responsible for numerous envenomations through their stinging organelles, nematocysts. These specialized microscopic structures discharge venom, yet detailed knowledge of their types [...] Read more.

The multiple-tentacle box jellyfish, Chironex indrasaksajiae (Sucharitakul, 2017) and Chiropsoides buitendijki (Horst, 1907), are venomous species found in Thai waters. They are responsible for numerous envenomations through their stinging organelles, nematocysts. These specialized microscopic structures discharge venom, yet detailed knowledge of their types and morphology in these species remains limited. This study updates the characterization of nematocyst types and features in C. indrasaksajiae and C. buitendijki using light and scanning electron microscopy for detailed examination. Four distinct nematocyst types were identified: banana-shaped microbasic p-mastigophores, oval-shaped microbasic p-rhopaloids, sub-spherical microbasic p-rhopaloids, and rod-shaped isorhizas. In C. indrasaksajiae, banana-shaped microbasic p-mastigophores exhibited significant intraspecific variability, ranging from 30.26 µm to 102.56 µm in length and 6.42 µm to 17.01 µm in width. Conversely, C. buitendijki showed a narrower size range, 72.17 µm to 98.37 µm in length and 10.73 µm to 16.48 µm in width, based on multiple individuals. The size ranges for the other nematocyst types were consistent across both species. This study enhances the understanding of nematocyst morphology in these box jellyfish, providing a foundation for further research on venom delivery mechanisms and improved management of jellyfish envenomations in Thai waters. Full article

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31 pages, 758 KiB

Open AccessReview

Aflatoxin B1: Challenges and Strategies for the Intestinal Microbiota and Intestinal Health of Monogastric Animals

by Hyunjun Choi, Yesid Garavito-Duarte, Alexa R. Gormley and Sung Woo Kim

Toxins 2025, 17(1), 43; https://doi.org/10.3390/toxins17010043 - 17 Jan 2025

Abstract

The objective of this review is to investigate the impacts of aflatoxins, particularly aflatoxin B1 (AFB₁), on intestinal microbiota, intestinal health, and growth performance in monogastric animals, primarily chickens and pigs, as well as dietary interventions to mitigate these effects. Aflatoxin [...] Read more.

The objective of this review is to investigate the impacts of aflatoxins, particularly aflatoxin B1 (AFB₁), on intestinal microbiota, intestinal health, and growth performance in monogastric animals, primarily chickens and pigs, as well as dietary interventions to mitigate these effects. Aflatoxin B1 contamination in feeds disrupts intestinal microbiota, induces immune responses and oxidative damage, increases antioxidant activity, and impairs jejunal cell viability, barrier function, and morphology in the small intestine. These changes compromise nutrient digestion and reduce growth performance in animals. The negative impact of AFB₁ on the % change in average daily gain (ΔADG) of chickens and pigs was estimated based on meta-analysis: ΔADG (%)_chicken = −0.13 × AFB₁ intake per body weight (ng/g·d) and ΔADG (%)_pig = −0.74 × AFB₁ intake per body weight (µg/kg·d), indicating that increasing AFB₁ contamination linearly reduces the growth of animals. To mitigate the harmful impacts of AFB₁, various dietary strategies have been effective. Mycotoxin-detoxifying agents include mycotoxin-adsorbing agents, such as clay and yeast cell wall compounds, binding to AFB₁ and mycotoxin-biotransforming agents, such as specific strains of Bacillus subtilis and mycotoxin-degrading enzyme, degrading AFB₁ into non-toxic metabolites such as aflatoxin D1. Multiple mycotoxin-detoxifying agents are often combined and used together to improve the intestinal health and growth of chickens and pigs fed AFB₁-contaminated feeds. In summary, AFB₁ negatively impacts intestinal microbiota, induces immune responses and oxidative stress, disrupts intestinal morphology, and impairs nutrient digestion in the small intestine, leading to reduced growth performance. Supplementing multi-component mycotoxin-detoxifying agents in feeds could effectively adsorb and degrade AFB₁ co-contaminated with other mycotoxins prior to its absorption in the small intestine, preventing its negative impacts on the intestinal health and growth performance of chickens and pigs. Full article

(This article belongs to the Special Issue Aspergillus flavus and Aflatoxins (Volume III))

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18 pages, 2085 KiB

Open AccessArticle

Crustacean Zooplankton Ingestion of Potentially Toxic Microcystis: In Situ Estimation Using mcyE Gene Gut Content Detection in a Large Temperate Eutrophic Lake

by Helen Agasild, Margarita Esmeralda Gonzales Ferraz, Madli Saat, Priit Zingel, Kai Piirsoo, Kätlin Blank, Veljo Kisand, Tiina Nõges and Kristel Panksep

Toxins 2025, 17(1), 42; https://doi.org/10.3390/toxins17010042 - 16 Jan 2025

Abstract

Grazing by zooplankton can regulate bloom-forming cyanobacteria but can also transfer toxin-producing cells, as well as toxic metabolites, to the food web. While laboratory investigations have provided extensive knowledge on zooplankton and toxic cyanobacteria interactions, information on zooplankton feeding on toxin-producing cyanobacteria in [...] Read more.

Grazing by zooplankton can regulate bloom-forming cyanobacteria but can also transfer toxin-producing cells, as well as toxic metabolites, to the food web. While laboratory investigations have provided extensive knowledge on zooplankton and toxic cyanobacteria interactions, information on zooplankton feeding on toxin-producing cyanobacteria in natural water bodies remains scarce. In this study, we quantified Microcystis-specific mcyE synthase genes from the gut contents of various cladoceran and copepod taxa to assess the in situ crustacean community and taxon-specific ingestion of potentially toxic Microcystis in Lake Peipsi, a large eutrophic lake in Estonia, Northern Europe. Microcystis cells with mcyE genes were found in all crustaceans examined. However, some species, such as the cyclopoid copepod Mesocyclops leuckarti, were more efficient in ingesting potentially toxic Microcystis than other co-occurring cladocerans (Daphnia spp., Bosmina spp., Chydorus sphaericus) and copepods (Eudiaptomus gracilis). The amount of toxigenic Microcystis cells grazed by crustacean population changed temporarily, and copepods were the predominant consumers of toxigenic Microcystis during several months of the 5-month study period. Crustacean ingestion of toxigenic Microcystis was not related to Microcystis biomass or mcyE gene copy numbers in the environment but was instead related to the abundance of major crustacean grazers. Our findings emphasize the close interaction between crustacean zooplankton and toxigenic Microcystis, indicating that some species may play a more significant role in linking toxic cells within the food web than others. Full article

(This article belongs to the Special Issue Harmful Algae in a Changing World: Where Did We Come from and Where Are We Going?)

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17 pages, 5733 KiB

Open AccessArticle

Spatial Distribution and Dietary Risk Assessment of Aflatoxins in Raw Milk and Dairy Feedstuff Samples from Different Climate Zones in China

by Xueli Yang, Bolin Liu, Lei Zhang, Xiaodan Wang, Jian Xie and Jiang Liang

Toxins 2025, 17(1), 41; https://doi.org/10.3390/toxins17010041 - 16 Jan 2025

Abstract

This study aimed to explore the contamination of aflatoxins by investigating the spatial distribution of aflatoxin B1 (AFB1) in cow feedstuff and aflatoxin M1 (AFM1) in raw milk, and the potential health risks of AFM1 in milk and dairy products. Feedstuff and raw [...] Read more.

This study aimed to explore the contamination of aflatoxins by investigating the spatial distribution of aflatoxin B1 (AFB1) in cow feedstuff and aflatoxin M1 (AFM1) in raw milk, and the potential health risks of AFM1 in milk and dairy products. Feedstuff and raw milk were collected from 160 pastures in three climate zones of China from October to November 2020. The results indicated the level of AFB1 and AFM1 ranged from 51.1 to 74.1 ng/kg and 3.0 to 7.0 ng/kg, respectively. Spatial analysis indicated the contamination was mostly concentrated in the temperate monsoon climate zone. On average, the estimated dietary exposure to AFM1 from milk and dairy products for Chinese consumers ranged from 0.0138 to 0.0281 ng/kg bw/day, with the MOE values below 10,000, and liver cancer risk of 0.00004–0.00009 cases/100,000 persons/year. For different groups, the average exposure to AFM1 was highest in the temperate monsoon climate zone and for toddlers. Full article

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Graphical abstract

20 pages, 2771 KiB

Open AccessArticle

Delphi Consensus on the Management of Spanish Patients with Post-Stroke Hemiplegic Shoulder Pain Treated with Botulinum Toxin A: Result Study

by Carlos Cordero-García, Irene de Torres, Jacobo Formigo-Couceiro, Lluis Guirao, Mª Dolores Romero-Torres, Sergio Otero-Villaverde, Alberto Herrera, Cristina Santa and Antonio Mena-Rodriguez

Toxins 2025, 17(1), 40; https://doi.org/10.3390/toxins17010040 - 16 Jan 2025

Abstract

The study aimed to identify expert opinions and obtain recommendations on the management of post-stroke hemiplegic shoulder pain (HSP) and treatment with botulinum toxin A (BoNT-A). A multicenter Delphi study was conducted using an online survey designed by a committee of experts with [...] Read more.

The study aimed to identify expert opinions and obtain recommendations on the management of post-stroke hemiplegic shoulder pain (HSP) and treatment with botulinum toxin A (BoNT-A). A multicenter Delphi study was conducted using an online survey designed by a committee of experts with at least 10 years of experience in post-stroke HSP management with BoNT-A in Spain. Forty-seven panelists (specialists with at least 5 years of experience in post-stroke HSP management with BoNT-A) rated their level of agreement in two rounds based on acceptance by ≥66.7% of them. In round 1, 245 statements on three dimensions were evaluated (diagnosis, treatment, and follow-up of the HSP patients treated with BoNT-A). A total of 159 statements (70.9%) were finally accepted after round 2. Experts recommended BoNT-A as soon as spasticity affects daily activities. They considered ultrasound as the preferred guided technique. Experts recommended regular assessments using validated scales and patient-reported outcomes to evaluate treatment goals and safety. In case of lack of response, experts suggested increasing the dose or number of treated muscles or considering alternative treatments. These consensus-based recommendations offer clinicians an approach to the management of post-stroke HSP with BoNT-A, supporting informed decision making. Full article

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18 pages, 1651 KiB

Open AccessArticle

Fusarium Head Blight in Argentina, a Profile of Produced Mycotoxins and a Biocontrol Strategy in Barley During Micro-Malting Process

by María Silvina Alaniz-Zanon, Marianela Bossa, Lorenzo Antonio Rosales Cavaglieri, Juan Manuel Palazzini, Michael Sulyok, Sofía Noemí Chulze and María Laura Chiotta

Toxins 2025, 17(1), 39; https://doi.org/10.3390/toxins17010039 - 15 Jan 2025

Abstract

Barley (Hordeum vulgare L.) is the second winter crop in Argentina. In the national market, grains are mainly destined to produce malt for beer manufacture. Fusarium species are common, causing Fusarium Head Blight (FHB) in barley, which generates yield and quality losses, [...] Read more.

Barley (Hordeum vulgare L.) is the second winter crop in Argentina. In the national market, grains are mainly destined to produce malt for beer manufacture. Fusarium species are common, causing Fusarium Head Blight (FHB) in barley, which generates yield and quality losses, as well as mycotoxin occurrence. The aims of this study were to determine (a) the incidence of the main species causing FHB in different locations of the barley-growing region of Argentina, (b) their ability to produce mycotoxins, and (c) the levels of deoxynivalenol (DON) and nivalenol (NIV) natural occurrence in grains at the harvest stage. Additionally, a strain of Bacillus velezensis was studied as a biocontrol agent in order to control F. graminearum sensu stricto and mycotoxin accumulation during the malting process, with the final objective being to reduce DON contamination in the beer manufacture chain. Fusarium graminearum ss was the most prevalent species causing FHB, with Fusarium poae being less distributed. Both species produced several mycotoxins, including NX-2 and NX-3, which is the first report of their production by strains isolated from barley in Argentina. Deoxynivalenol contamination was found in 95% of barley grains during the 2016 harvest season (mean: 0.4 mg/kg), while NIV contamination was present in 29% of samples (mean: 0.49 mg/kg). In the 2017 harvest season, 53.6% of grains were contaminated with DON (mean: 0.42 mg/kg), and 21% with NIV (mean: 0.8 mg/kg). Quantification of F. graminearum ss by real-time PCR during the micro-malting process showed that application of the biocontrol agent before the germination stage was the most effective treatment, with a 45% reduction in fungal DNA levels. Reduction in DON contamination (69.3–100%) in artificially infected grains with F. graminearum ss, was also observed. The present work contributes to the knowledge of FHB in Argentina and to the development of a strategy to control this disease and mycotoxin contamination in barley, promoting at the same time food security. Full article

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

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13 pages, 3637 KiB

Open AccessArticle

A Study of Combined Onabotulinumtoxin A and Hyaluronic Acid Filler for the Treatment of Enlarged Facial Pores

by Vasanop Vachiramon, Sonphet Chirasuthat, Suphagan Boonpethkaew, Nawara Sakpuwadol, Tanat Yongpisarn and Natthachat Jurairattanaporn

Toxins 2025, 17(1), 38; https://doi.org/10.3390/toxins17010038 - 15 Jan 2025

Abstract

Introduction: Enlarged facial pores are a common cosmetic concern caused by excessive sebum production, visible hair shafts, and a reduction in skin elasticity, leading to a decrease in skin quality and overall appearance. Various treatment modalities have been explored to address this issue. [...] Read more.

Introduction: Enlarged facial pores are a common cosmetic concern caused by excessive sebum production, visible hair shafts, and a reduction in skin elasticity, leading to a decrease in skin quality and overall appearance. Various treatment modalities have been explored to address this issue. This study focuses on the efficacy and safety of combining Onabotulinumtoxin A (OnaBoNT-A) and hyaluronic acid filler (HA filler) to target enlarged facial pores in Asians. Materials and Methods: This study aimed to compare the efficacy and safety of OnaBoNT-A monotherapy in combination with HA filler for the treatment of enlarged facial pores. This study was a prospective, randomized, single-blinded, split-face, controlled trial that enrolled 32 subjects with visibly enlarged pores on both cheeks. One side of the face received intradermal injections of OnaBoNT-A, while the other side received OnaBoNT-A in combination with intradermal hyaluronic acid filler injection. The outcomes were measured by pore volume, visual assessment, pain score, improvement score, and side effects at various time intervals up to 24 weeks. Results: This study investigated the effects of onaBoNT-A monotherapy or in combination with HA filler on facial pore size and skin roughness. The results showed that both sides exhibited a reduction in pore volume and skin roughness over time, but the side treated with onaBoNT-A monotherapy had a slightly better improvement than the combination side at the 6-month follow-up. Subjects with histories of facial oiliness were more likely to respond to onaBoNT-A monotherapy, while those without histories of facial oiliness were more likely to respond to the side treated with combined treatment. The most common adverse events were erythema, bruising, and edema, which were more frequent on the combination side. Additionally, 18 subjects (56.25%) experienced a palpable lump on the combination side, which resolved in most cases within a few months. Conclusion: BoNT-A and HA dermal filler had a role in reducing pore size. Nonetheless, individuals with enlarged pores who exhibited beneficial effects to botulinum toxin injection typically had a background of facial oiliness. Adverse incidents like dermal edema and palpable nodules were observed, underscoring the significance of meticulous patient selection and accurate injection technique. Full article

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15 pages, 1275 KiB

Open AccessArticle

Evaluating Methods for Aflatoxin B1 Monitoring in Selected Food Crops Within Decentralized Agricultural Systems

by Haadia Tanveer, Hannah Glesener, Blake Su, Brooke Bolsinger, Rosa Krajmalnik-Brown and Lee E. Voth-Gaeddert

Toxins 2025, 17(1), 37; https://doi.org/10.3390/toxins17010037 - 14 Jan 2025

Abstract

Aflatoxin B1 (AFB1) contamination of food crops pose severe public health risks, particularly in decentralized agricultural systems common in low-resource settings. Effective monitoring tools are critical for mitigating exposure, but their adoption is limited by barriers such as cost, infrastructure, and technical expertise. [...] Read more.

Aflatoxin B1 (AFB1) contamination of food crops pose severe public health risks, particularly in decentralized agricultural systems common in low-resource settings. Effective monitoring tools are critical for mitigating exposure, but their adoption is limited by barriers such as cost, infrastructure, and technical expertise. The objectives of this study were: (1) to evaluate common AFB1 detection methods, including enzyme-linked immunosorbent assays (ELISA) and lateral-flow assays (LFA), validated via high-performance liquid chromatography (HPLC), focusing on their suitability for possible applications in decentralized, low-resource settings; and (2) to conduct a barriers-to-use assessment for commonly available AFB1 detection methods and their applicability in low-resource settings. Among four ELISA kits, the AgraQuant Aflatoxin B1 2/50 ELISA Kit demonstrated the highest accuracy and precision, reliably quantifying AFB1 in maize and tortillas across 5–150 ppb with minimal cross-reactivity. For LFA, a smartphone-based algorithm achieved a high presence/absence accuracy rate of 84% but struggled with concentration prediction. The barriers-to-use analysis highlighted the practicality of low-cost tools like moisture readers for field screening but underscored their qualitative limitations. Advanced methods like HPLC and LC-MS offer greater precision but remain impractical due to their high costs and infrastructure requirements, suggesting a potential role for adapted ELISA or LFA methods as confirmatory approaches. These findings support the development of multi-tiered frameworks integrating affordable field tools with regional or centralized confirmatory testing. Addressing systemic barriers through capacity building, partnerships, and improved logistics will enhance AFB1 monitoring in decentralized systems, protecting public health in vulnerable communities. Full article

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22 pages, 3875 KiB

Open AccessReview

Venoms and Extracellular Vesicles: A New Frontier in Venom Biology

by Auwal A. Bala, Naoual Oukkache, Elda E. Sanchez, Montamas Suntravat and Jacob A. Galan

Toxins 2025, 17(1), 36; https://doi.org/10.3390/toxins17010036 - 14 Jan 2025

Abstract

Extracellular vesicles (EVs) are nanoparticle-sized vesicles secreted by nearly all cell types under normal physiological conditions. In toxicological research, EVs have emerged as a crucial link between public health and multi-omics approaches, offering insights into cellular responses to disease-causing injury agents such as [...] Read more.

Extracellular vesicles (EVs) are nanoparticle-sized vesicles secreted by nearly all cell types under normal physiological conditions. In toxicological research, EVs have emerged as a crucial link between public health and multi-omics approaches, offering insights into cellular responses to disease-causing injury agents such as environmental and biological toxins, contaminants, and drugs. Notably, EVs present a unique opportunity to deepen our understanding of the pathophysiology of envenomation by natural toxins. Recent advancements in isolating and purifying EV cargo, mass spectrometry techniques, and bioinformatics have positioned EVs as potential biomarkers that could elucidate biological signaling pathways and provide valuable information on the relationship between venomous toxins, their mechanisms of action, and the effectiveness of antivenoms. Additionally, EVs hold promise as proxies for various aspects of envenomation, including the toxin dosage, biological characterization, injury progression, and prognosis during therapeutic interventions. These aspects can be explored through multi-omics technology applied to EV contents from the plasma, saliva, or urine samples of envenomated individuals, offering a comprehensive integrative approach to understanding and managing envenomation cases. Full article

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

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16 pages, 905 KiB

Open AccessArticle

Risk Assessment of Harmful Algal Blooms in Salmon Farming: Scotland as a Case Study

by Fatima Gianella, Michael T. Burrows and Keith Davidson

Toxins 2025, 17(1), 35; https://doi.org/10.3390/toxins17010035 - 13 Jan 2025

Abstract

This study explored harmful algal bloom (HAB) risk as a function of exposure, hazard and vulnerability, using Scotland as a case study. Exposure was defined as the fish biomass estimated to be lost from a bloom event, based on the total recorded annual [...] Read more.

This study explored harmful algal bloom (HAB) risk as a function of exposure, hazard and vulnerability, using Scotland as a case study. Exposure was defined as the fish biomass estimated to be lost from a bloom event, based on the total recorded annual production. Hazard was estimated from literature-reported bloom events. Vulnerability was calculated from records of the number of employees (2020), as an estimate of aquaculture-based employment. The dinoflagellate Karenia mikimotoi was identified as the HAB species with the highest frequency of reported bloom events in Scotland, with variable spatial and temporal reports, but environmental and climatological variables regulating these events are currently unknown. The Shetland Islands region exhibited the highest combined HAB risk, with the highest scores in all three components. Vulnerability was particularly important to overall risk within an island setting, where a larger proportion of the population was dependent on aquaculture. The analysis demonstrated the potential to evaluate the economic and social consequences of HAB events on the aquaculture industry. As fish-killing HABs and fish health impacts are likely under-reported, more transparent reporting of events and related fish health and physiological consequences is recommended for a more quantitative application of this approach. Full article

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20 pages, 1431 KiB

Open AccessArticle

Mapping Variability of Mycotoxins in Individual Oat Kernels from Batch Samples: Implications for Sampling and Food Safety

by Irene Teixido-Orries, Francisco Molino, Bianca Castro-Criado, Monika Jodkowska, Angel Medina, Sonia Marín and Carol Verheecke-Vaessen

Toxins 2025, 17(1), 34; https://doi.org/10.3390/toxins17010034 - 11 Jan 2025

Abstract

Oats are susceptible to contamination by Fusarium mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), and T-2/HT-2 toxins, posing food safety risks. This study analyses the variation in levels of 14 mycotoxins in 200 individual oat kernels from two DON-contaminated batch samples (mean = 3498 [...] Read more.

Oats are susceptible to contamination by Fusarium mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), and T-2/HT-2 toxins, posing food safety risks. This study analyses the variation in levels of 14 mycotoxins in 200 individual oat kernels from two DON-contaminated batch samples (mean = 3498 µg/kg) using LC-MS/MS. The samples also contained deoxynivalenol-3-glucoside (DON-3G), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and ZEN. Contamination levels varied notably among individual kernels, with DON detected in 70% of them, followed by DON-3G (24.5%) and 3-ADON (20.5%). Importantly, 8% of kernels exceeded the EU legal limit for DON (1750 µg/kg), and some occasionally surpassed limits for ZEN and T-2/HT-2. Correlation analyses revealed strong associations between DON and its derivatives but weaker correlations with other toxins. Mycotoxin ratios varied widely, indicating that although they often co-occur, their concentrations differ between kernels. Contamination did not significantly impact kernel weight, though a slight trend toward lower weights in contaminated kernels was noted. Additionally, sampling statistics showed that as the percentage of selected kernels increased, the probability of batch sample rejection for DON contamination rose significantly. The study highlights the heterogeneity of mycotoxin contamination in oat batches, emphasising the importance of accurate detection and regulatory compliance to ensure safer oat-based products. Full article

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

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17 pages, 5927 KiB

Open AccessArticle

Pulsed Electric Field Induces Significant Changes in the Metabolome of Fusarium Species and Decreases Their Viability and Toxigenicity

by Adam Behner, Jana Palicova, Anna-Hirt Tobolkova, Nela Prusova and Milena Stranska

Toxins 2025, 17(1), 33; https://doi.org/10.3390/toxins17010033 - 11 Jan 2025

Abstract

Fusarium fungi are widespread pathogens of food crops, primarily associated with the formation of mycotoxins. Therefore, effective mitigation strategies for these toxicogenic microorganisms are required. In this study, the potential of pulsed electric field (PEF) as an advanced technology of increasing use in [...] Read more.

Fusarium fungi are widespread pathogens of food crops, primarily associated with the formation of mycotoxins. Therefore, effective mitigation strategies for these toxicogenic microorganisms are required. In this study, the potential of pulsed electric field (PEF) as an advanced technology of increasing use in the food processing industry was investigated to minimize the viability of Fusarium pathogens and to characterize the PEF-induced changes at the metabolomic level. Spores of four Fusarium species (Fusarium culmorum, Fusarium graminearum, Fusarium poae, and Fusarium sporotrichioides) were treated with PEF and cultured on potato dextrose agar (PDA) plates. The viability of the Fusarium species was assessed by counting the colony-forming units, and changes in the mycotoxin content and metabolomic fingerprints were evaluated by using UHPLC-HRMS/MS instrumental analysis. For metabolomic data processing and compound identification, the MS-DIAL (v. 4.80)–MS-CleanR–MS-Finder (v. 3.52) software platform was used. As we found out, both fungal viability and the ability to produce mycotoxins significantly decreased after the PEF treatment for all of the species tested. The metabolomes of the treated and untreated fungi showed statistically significant differences, and PEF-associated biomarkers from the classes oxidized fatty acid derivatives, cyclic hexapeptides, macrolides, pyranocoumarins, carbazoles, and guanidines were identified. Full article

(This article belongs to the Section Mycotoxins)

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13 pages, 1661 KiB

Open AccessArticle

Encapsulated LyeTx III Peptide: Cytotoxic Agent Isolated from Lycosa erythrognatha Spider Venom

by Daniel Moreira dos Santos, Livia Ramos Santiago, Nayara Araújo dos Santos, Wanderson Romão, Jarbas Magalhães Resende, Maria Elena de Lima, Márcia Helena Borges and Rosy Iara Maciel de Azambuja Ribeiro

Toxins 2025, 17(1), 32; https://doi.org/10.3390/toxins17010032 - 10 Jan 2025

Abstract

The discovery of novel cytotoxic drugs is of paramount importance in contemporary medical research, particularly in the search for treatments with fewer side effects and higher specificity. Antimicrobial peptides are an interesting class of molecules for this endeavor. In this context, the LyeTx [...] Read more.

The discovery of novel cytotoxic drugs is of paramount importance in contemporary medical research, particularly in the search for treatments with fewer side effects and higher specificity. Antimicrobial peptides are an interesting class of molecules for this endeavor. In this context, the LyeTx III, a new peptide extracted from the venom of the Lycosa erythrognatha spider, stands out. The peptide exhibits typical antimicrobial traits: a positive net charge and amphipathic α -helix structure in lipid-like environments. Its unique sequence (GKAMKAIAKFLGR-NH₂), identified via mass spectrometry and Edman degradation, shows limited similarity to existing peptides. Significantly, when liposome-encapsulated, LyeTx III demonstrates selective activity against tumor cells in culture. Our MTT results showed that the cytotoxicity of the peptide increased against HN13 cells when administered as liposomes, with their viability in HN13 cells alone being 98%, compared to 38% in liposome-encapsulated form. This finding underscores that the LyeTx III peptide may be a good candidate for the development of new drugs against cancer. Its activity when encapsulated is promising, as it can increase its half-life in the body and can also be targeted to specific tumors. Full article

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

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12 pages, 290 KiB

Open AccessReview

The Esthetic Use of Botulinum Toxins in Cancer Patients: Providing a Foundation for Future Indications

by Marco Papagni, Monica Renga, Selene Mogavero, Paolo Veronesi and Maurizio Cavallini

Toxins 2025, 17(1), 31; https://doi.org/10.3390/toxins17010031 - 10 Jan 2025

Abstract

Advances in oncological treatments have improved the survival rates of cancer patients but have often resulted in significant physical changes that negatively impact their self-esteem and psychological well-being. Cancer patients frequently ask esthetic practitioners to perform procedures to address such changes. However, practitioners [...] Read more.

Advances in oncological treatments have improved the survival rates of cancer patients but have often resulted in significant physical changes that negatively impact their self-esteem and psychological well-being. Cancer patients frequently ask esthetic practitioners to perform procedures to address such changes. However, practitioners often hesitate to satisfy such requests due to lacking guidelines or recommendations. The use of botulinum toxins (BoNTs) for esthetic purposes has shown significant promise in improving the quality of life for cancer patients. This review explores the broad application of BoNTs in many medical branches, focusing on oncology. A substantial amount of literature shows that BoNTs are safe and effective as a type of adjunctive therapy compared to classical cancer treatments. We provide our expert opinion that the use of BoNTs for esthetic purposes is safe for cancer patients and even recommended for those whose mood is influenced by the worsening of their physical appearance. Careful patient selection and interdisciplinary collaboration are essential to the safe integration of BoNTs into cancer care. Full article

(This article belongs to the Section Bacterial Toxins)

44 pages, 3456 KiB

Open AccessReview

Species Differences in the Biotransformation of Aflatoxin B1: Primary Determinants of Relative Carcinogenic Potency in Different Animal Species

by David L. Eaton, David E. Williams and Roger A. Coulombe

Toxins 2025, 17(1), 30; https://doi.org/10.3390/toxins17010030 - 9 Jan 2025

Abstract

It has been known since the early days of the discovery of aflatoxin B1 (AFB1) that there were large species differences in susceptibility to AFB1. It was also evident early on that AFB1 itself was not toxic but required bioactivation to a reactive [...] Read more.

It has been known since the early days of the discovery of aflatoxin B1 (AFB1) that there were large species differences in susceptibility to AFB1. It was also evident early on that AFB1 itself was not toxic but required bioactivation to a reactive form. Over the past 60 years there have been thousands of studies to delineate the role of ~10 specific biotransformation pathways of AFB1, both phase I (oxidation, reduction) and phase II (hydrolysis, conjugation, secondary oxidations, and reductions of phase I metabolites). This review provides a historical context and substantive analysis of each of these pathways as contributors to species differences in AFB1 hepatoxicity and carcinogenicity. Since the discovery of AFB1 as the toxic contaminant in groundnut meal that led to Turkey X diseases in 1960, there have been over 15,000 publications related to aflatoxins, of which nearly 8000 have addressed the significance of biotransformation (metabolism, in the older literature) of AFB1. While it is impossible to give justice to all of these studies, this review provides a historical perspective on the major discoveries related to species differences in the biotransformation of AFB1 and sets the stage for discussion of other papers in this Special Issue of the important role that AFB1 metabolites have played as biomarkers of exposure and effect in thousands of human studies on the toxic effects of aflatoxins. Dr. John Groopman has played a leading role in every step of the way—from initial laboratory studies on specific AFB1 metabolites to the application of molecular biomarkers in epidemiological studies associating dietary AFB1 exposure with liver cancer, and the design and conduct of chemoprevention clinical trials to reduce cancer risk from unavoidable aflatoxin exposures by alteration of specific AFB1 biotransformation pathways. This article is written in honor of Dr. Groopman’s many contributions in this area. Full article

(This article belongs to the Special Issue 65 Years On from Aflatoxin Discovery—a Themed Issue in Honor of Professor John D. Groopman)

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18 pages, 4143 KiB

Open AccessArticle

Proteomic Analysis of the Murine Liver Response to Oral Exposure to Aflatoxin B1 and Ochratoxin A: The Protective Role to Bioactive Compounds

by Silvia Trombetti, Alessandra Cimbalo, Michela Grosso, Pilar Vila-Donat, Jordi Mañes and Lara Manyes

Toxins 2025, 17(1), 29; https://doi.org/10.3390/toxins17010029 - 9 Jan 2025

Abstract

Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are considered the most important mycotoxins in terms of food safety. The aim of this study was to evaluate the hepatotoxicity of AFB1 and OTA exposure in Wistar rats and to assess the beneficial effect of [...] Read more.

Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are considered the most important mycotoxins in terms of food safety. The aim of this study was to evaluate the hepatotoxicity of AFB1 and OTA exposure in Wistar rats and to assess the beneficial effect of fermented whey (FW) and pumpkin (P) as functional ingredients through a proteomic approach. For the experimental procedures, rats were fed AFB1 and OTA individually or in combination, with the addition of FW or a FW-P mixture during 28 days. For proteomics analysis, peptides were separated using a LC-MS/MS-QTOF system and differentially expressed proteins (DEPs) were statistically filtered (p < 0.05) distinguishing males from females. Gene ontology visualization allowed the identification of proteins involved in important biological processes such as the response to xenobiotic stimuli and liver development. Likewise, KEGG pathway analysis reported the metabolic routes as the most affected, followed by carbon metabolism and biosynthesis of amino acids. Overall, the results highlighted a strong downregulation of DEPs in the presence of AFB1 and OTA individually but not with the mixture of both, suggesting a synergistic effect. However, FW and P have helped in the mitigation of processes triggered by mycotoxins. Full article

(This article belongs to the Special Issue Novel Approaches in Mycotoxins Research: Detection, Prevention and Mode of Action)

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10 pages, 1053 KiB

Open AccessArticle

Isolation and Structural Identification of New Diol Esters of Okadaic Acid and Dinophysistoxin-1 from the Cultured Prorocentrum lima

by Yeong Kwang Ji, Semin Moon, Sangbum Lee, Yun Na Kim, Eun Ju Jeong and Jung-Rae Rho

Toxins 2025, 17(1), 28; https://doi.org/10.3390/toxins17010028 - 7 Jan 2025

Abstract

Prorocentrum, a dinoflagellate responsible for producing diarrhetic shellfish poisoning (DSP) toxins, poses significant threats to marine ecosystems, aquaculture industries, and human health. DSP toxins, including okadaic acid (OA), dinophysis toxin (DTX), and their diverse derivatives, continue to be identified and characterized. In [...] Read more.

Prorocentrum, a dinoflagellate responsible for producing diarrhetic shellfish poisoning (DSP) toxins, poses significant threats to marine ecosystems, aquaculture industries, and human health. DSP toxins, including okadaic acid (OA), dinophysis toxin (DTX), and their diverse derivatives, continue to be identified and characterized. In this study, we report the isolation of four new diol esters of OA/DTX-1 from large-scale cultures of Prorocentrum lima. Their chemical structures were elucidated through comprehensive NMR and MS analyses, along with structural comparisons with the well-known OA. Notably, compound 1 featured an additional ester group within the diol unit, while compound 2 was revealed to be a C11 diol ester. The cytotoxicity of these newly isolated derivatives was evaluated against three cell lines: Neuro2a (mouse), HCT116 (human), and HepG2 (human). All diol esters exhibited cytotoxic effects, with compound 3 displaying toxicity comparable to OA. These results expand our understanding of DSP toxin diversity and provide valuable insight into the structural variations and biological activity of diol esters of OA/DTX-1. Full article

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