Toxins

12 pages, 1689 KiB

Open AccessArticle

Insights into the Ecotoxicology of Radicinin and (10S,11S)-(—)-epi-Pyriculol, Fungal Metabolites with Potential Application for Buffelgrass (Cenchrus ciliaris) Biocontrol

by Antonietta Siciliano, Jesús G. Zorrilla, Lorenzo Saviano, Alessio Cimmino, Marco Guida, Marco Masi and Susan Meyer

Toxins 2023, 15(6), 405; https://doi.org/10.3390/toxins15060405 - 20 Jun 2023

Cited by 1 | Viewed by 937

Abstract

Buffelgrass (Cenchrus ciliaris L.) is an invasive C4 perennial grass species that substantially reduces native plant diversity of the Sonoran Desert through fire promotion and resource competition. Broad-spectrum herbicides are essentially used for its control, but they have a negative environmental and [...] Read more.

Buffelgrass (Cenchrus ciliaris L.) is an invasive C4 perennial grass species that substantially reduces native plant diversity of the Sonoran Desert through fire promotion and resource competition. Broad-spectrum herbicides are essentially used for its control, but they have a negative environmental and ecological impact. Recently, phytotoxicity on C. ciliaris has been discovered for two metabolites produced in vitro by the phytopathogenic fungi Cochliobolus australiensis and Pyricularia grisea. They were identified as (10S,11S)-(—)-epi-pyriculol and radicinin and resulted in being potential candidates for the development of bioherbicides for buffelgrass biocontrol. They have already shown promising results, but their ecotoxicological profiles and degradability have been poorly investigated. In this study, ecotoxicological tests against representative organisms from aquatic ecosystems (Aliivibrio fischeri bacterium, Raphidocelis subcapitata alga, and Daphnia magna crustacean) revealed relatively low toxicity for these compounds, supporting further studies for their practical application. The stability of these metabolites in International Organization for Standardization (ISO) 8692:2012 culture medium under different temperatures and light conditions was also evaluated, revealing that 98.90% of radicinin degraded after 3 days in sunlight. Significant degradation percentages (59.51–73.82%) were also obtained at room temperature, 30 °C or under ultraviolet (254 nm) light exposure. On the other hand, (10S,11S)-epi-pyriculol showed more stability under all the aforementioned conditions (49.26–65.32%). The sunlight treatment was also shown to be most effective for the degradation of this metabolite. These results suggest that radicinin could provide rapid degradability when used in agrochemical formulations, whereas (10S,11S)-epi-pyriculol stands as a notably more stable compound. Full article

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13 pages, 6348 KiB

Open AccessArticle

Microcystin-LR-Exposure-Induced Kidney Damage by Inhibiting MKK6-Mediated Mitophagy in Mice

by Xueqiong Yao, Ying Liu, Yue Yang, Yafang Li, Na Hu, Fengmei Song and Fei Yang

Toxins 2023, 15(6), 404; https://doi.org/10.3390/toxins15060404 - 19 Jun 2023

Cited by 3 | Viewed by 1443

Abstract

Previous studies have reported that microcystin-LR (MC-LR) levels are highly correlated with abnormal renal function indicators, suggesting that MC-LR is an independent risk factor for kidney damage. However, the evidence for the exact regulation mechanism of MC-LR on kidney damage is still limited, [...] Read more.

Previous studies have reported that microcystin-LR (MC-LR) levels are highly correlated with abnormal renal function indicators, suggesting that MC-LR is an independent risk factor for kidney damage. However, the evidence for the exact regulation mechanism of MC-LR on kidney damage is still limited, and further in-depth exploration is needed. In addition, the mitochondria-related mechanism of MC-LR leading to kidney damage has not been elucidated. To this end, the present study aimed to further explore the mechanism of mitophagy related to kidney damage induced by MC-LR through in vitro and in vivo experiments. Male C57BL/6 mice were fed with a standard rodent pellet and exposed daily to MC-LR (20 μg/kg·bw) via intraperitoneal injections for 7 days. Moreover, HEK 293 cells were treated with MC-LR (20 μM) for 24 h. The histopathological results exhibited kidney damage after MC-LR exposure, characterized by structurally damaged nephrotomies, with inflammatory cell infiltration. Similarly, a significant increase in renal interstitial fibrosis was observed in the kidneys of MC-LR-treated mice compared with those of the control group (CT) mice. MC-LR exposure caused impaired kidney function, with markedly increased blood urea nitrogen (BUN), creatinine (Cr), and uric acid (UA) levels in mice. Ultrastructural analysis exhibited obviously swollen, broken, and disappearing mitochondrial crests, and partial mitochondrial vacuoles in the MC-LR-treated HEK 293 cells. The Western blotting results demonstrated that exposure to MC-LR significantly increased the protein expressions of MKK6, p-p38, and p62, while the expression of mitophagy-related proteins was significantly inhibited in the kidneys of mice and HEK293 cells, including parkin, TOM20, and LC3-II, indicating the inhibition of mitophagy. Therefore, our data suggest that the inhibition of MKK6-mediated mitophagy might be the toxicological mechanism of kidney toxicity in mice with acute exposure to MC-LR. Full article

(This article belongs to the Special Issue Recent Advances in Microcystins)

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39 pages, 5344 KiB

Open AccessReview

Insights into Toxic Prymnesium parvum Blooms as a Cause of the Ecological Disaster on the Odra River

by Janusz Sobieraj and Dominik Metelski

Toxins 2023, 15(6), 403; https://doi.org/10.3390/toxins15060403 - 19 Jun 2023

Cited by 3 | Viewed by 2477

Abstract

In 2022, Poland and Germany experienced a prolonged and extensive mass fish kill in the Odra River. During the period from the end of July to the beginning of September 2022, a high level of incidental disease and mortality was observed in various [...] Read more.

In 2022, Poland and Germany experienced a prolonged and extensive mass fish kill in the Odra River. During the period from the end of July to the beginning of September 2022, a high level of incidental disease and mortality was observed in various fish species (dozens of different species were found dead). The fish mortality affected five Polish provinces (Silesia, Opole, Lower Silesia, Lubuskie, and Western Pomerania) and involved reservoir systems covering most of the river (the Odra River is 854 km long, of which 742 km are in Poland). Fatal cases were investigated using toxicological, anatomopathological, and histopathological tests. Water samples were collected to determine nutrient status in the water column, phytoplankton biomass, and community composition. High nutrient concentrations indicated high phytoplankton productivity, with favorable conditions for golden algal blooms. The harmful toxins (prymnesins secreted by Prymnesium parvum habitats) had not been found in Poland before, but it was only a matter of time, especially in the Odra River, whose waters are permanently saline and still used for navigation. The observed fish mortality resulted in a 50% decrease in the fish population in the river and affected mainly cold-blooded species. Histopathological examinations of fish showed acute damage to the most perfused organs (gills, spleen, kidneys). The disruption to hematopoietic processes and damage to the gills were due to the action of hemolytic toxins (prymnesins). An evaluation of the collected hydrological, meteorological, biological, and physico-chemical data on the observed spatio-temporal course of the catastrophe, as well as the detection of three compounds from the group of B-type prymnesins in the analyzed material (the presence of prymnesins was confirmed using an analysis of the fragmentation spectrum and the accurate tandem mass spectrometry (MS/MS) measurement, in combination with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS), allowed the formulation and subsequent testing of the hypothesis for a direct link between the observed fish mortality and the presence of prymnesins in the Odra River. This article systematizes what is known about the causes of the fish kill in the Odra River in 2022, based on official government reports (one Polish and one German) and the EU technical report by the Joint Research Centre. A review and critical analysis of government findings (Polish and German) on this disaster were conducted in the context of what is known to date about similar cases of mass fish kills. Full article

(This article belongs to the Special Issue Unveiling the Toxic Effects of Harmful Algal Blooms)

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17 pages, 2507 KiB

Open AccessArticle

Biocontrol Potential of Antagonistic Yeasts on In Vitro and In Vivo Aspergillus Growth and Its AFB₁ Production

by Dilara Nur Dikmetas, Hayrettin Özer and Funda Karbancıoglu-Guler

Toxins 2023, 15(6), 402; https://doi.org/10.3390/toxins15060402 - 19 Jun 2023

Cited by 2 | Viewed by 1406

Abstract

Aspergillus flavus is a major aflatoxin B₁, posing significant health concerns to humans, crops, and producer fungi. Due to the undesirable consequences of the usage of synthetic fungicides, biological control using yeasts has gained more attention. In this study, eight isolates [...] Read more.

Aspergillus flavus is a major aflatoxin B₁, posing significant health concerns to humans, crops, and producer fungi. Due to the undesirable consequences of the usage of synthetic fungicides, biological control using yeasts has gained more attention. In this study, eight isolates of epiphytic yeasts belonging to Moesziomyces sp., Meyerozyma sp. and Metschnikowia sp., which have been identified as antagonists, were isolated from different plants, including grapes, blueberries, hawthorns, hoşkıran, beans and grape leaf. While volatile organic compounds (VOCs) produced by Moesziomyces bullatus DN-FY, Metschnikowia aff. pulcherrima DN-MP and Metschnikowia aff. pulcherrima 32-AMM reduced in vitro A. flavus mycelial growth and sporulation, only VOCs produced by Metschnikowia aff. fructicola 1-UDM were found to be effective at reducing in vitro AFB₁ production. All yeasts reduced the mycelial growth of A. flavus by 76–91%, while AFB₁ production reduced to 1.26–10.15 ng/g and the control plates’ growth was 1773 ng/g. The most effective yeast, Metschnikowia aff. Pulcherrima DN-HS, reduced Aspergillus flavus growth and aflatoxin B₁ production on hazelnuts. The AFB₁ content on hazelnuts reduced to 333.01 ng/g from 536.74 ng/g. To our knowledge, this is the first report of yeasts isolated from plants being tested as potential biological control agents to reduce AFB₁ production on hazelnuts. Full article

(This article belongs to the Special Issue Impact of Host-Fungi-Environment Interaction on Parent and Modified Mycotoxin Co-occurrence)

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12 pages, 933 KiB

Open AccessArticle

Simultaneous Determination of Pyrethrins, Pyrethroids, and Piperonyl Butoxide in Animal Feeds by Liquid Chromatography-Tandem Mass Spectrometry

by Xin Xu and Lisa A. Murphy

Toxins 2023, 15(6), 401; https://doi.org/10.3390/toxins15060401 - 17 Jun 2023

Cited by 1 | Viewed by 1567

Abstract

The presence of insecticides like pyrethrins and synthetic pyrethroids, combined with the synergist piperonyl butoxide, in animal feeds can pose a risk to both animal and human health by contaminating the food chain. In this study, a simple and fast method was developed [...] Read more.

The presence of insecticides like pyrethrins and synthetic pyrethroids, combined with the synergist piperonyl butoxide, in animal feeds can pose a risk to both animal and human health by contaminating the food chain. In this study, a simple and fast method was developed for the simultaneous determination of these compounds in contaminated animal feeds using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Sample preparation was carried out using a QuEChERS-based approach, and the method was validated with acceptable accuracy ranging from 84 to 115% and precision below 10%. The limit of detection (LOD) and limit of quantification (LOQ) were between 0.15 and 3 and 1 and 10 µg/kg, respectively. The method detected insecticide contaminations in various livestock and poultry feeds. Furthermore, the method was applied to a toxicology case, where it identified and quantified piperonyl butoxide and deltamethrin in the submitted horse feed sample. These results demonstrate that the method can be a valuable tool in animal health and food safety diagnostic applications, as well as veterinary toxicology investigations concerning pyrethrin-related feed contamination. Full article

(This article belongs to the Special Issue Animal Poisoning: Toxins from Plants or Feed)

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14 pages, 1353 KiB

Open AccessArticle

Development of Thermally Stable Nanobodies for Detection and Neutralization of Staphylococcal Enterotoxin B

by Anna C. Hughes, Marina Kirkland, Wenxian Du, Reuven Rasooly, Bradley Hernlem, Christina Tam, Yuzhu Zhang and Xiaohua He

Toxins 2023, 15(6), 400; https://doi.org/10.3390/toxins15060400 - 16 Jun 2023

Viewed by 1398

Abstract

In this study, sixteen unique staphylococcal enterotoxin B (SEB)-reactive nanobodies (nbs), including ten monovalent and six bivalent nbs, were developed. All characterized nbs were highly specific for SEB and did not cross-react with other staphylococcal enterotoxins (SE). Several formats of highly sensitive enzyme-linked [...] Read more.

In this study, sixteen unique staphylococcal enterotoxin B (SEB)-reactive nanobodies (nbs), including ten monovalent and six bivalent nbs, were developed. All characterized nbs were highly specific for SEB and did not cross-react with other staphylococcal enterotoxins (SE). Several formats of highly sensitive enzyme-linked immunosorbent assays (ELISAs) were established using SEB nbs and a polyclonal antibody (pAb). The lowest limit of detection (LOD) reached 50 pg/mL in PBS. When applied to an ELISA to detect SEB-spiked milk (a commonly contaminated foodstuff), a LOD as low as 190 pg/mL was obtained. The sensitivity of ELISA was found to increase concurrently with the valency of nbs used in the assay. In addition, a wide range of thermal tolerance was observed among the sixteen nbs, with a subset of nbs, SEB-5, SEB-9, and SEB-6², retaining activity even after exposure to 95 °C for 10 min, whereas the conventional monoclonal and polyclonal antibodies exhibited heat-labile properties. Several nbs demonstrated a long shelf-life, with one nb (SEB-9) retaining 93% of its activity after two weeks of storage at room temperature. In addition to their usage in toxin detection, eleven out of fifteen nbs were capable of neutralizing SEB’s super-antigenic activity, demonstrated by their inhibition on IL-2 expression in an ex vivo human PBMC assay. Compared to monoclonal and polyclonal antibodies, the nbs are relatively small, thermally stable, and easy to produce, making them useful in applications for sensitive, specific, and cost-effective detection and management of SEB contamination in food products. Full article

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1 pages, 177 KiB

Open AccessCorrection

Correction: Tsai et al. The Efficacy of Transvaginal Ultrasound-Guided BoNT-A External Sphincter Injection in Female Patients with Underactive Bladder. Toxins 2023, 15, 199

by Cheng-Yen Tsai, Yao-Hung Yeh, Li-Hsien Tsai and Eric Chieh-Lung Chou

Toxins 2023, 15(6), 399; https://doi.org/10.3390/toxins15060399 - 16 Jun 2023

Viewed by 945

Abstract

In the original publication [...] Full article

23 pages, 3384 KiB

Open AccessFeature PaperReview

Snake Antivenoms—Toward Better Understanding of the Administration Route

by Erika Gamulin, Sanja Mateljak Lukačević, Beata Halassy and Tihana Kurtović

Toxins 2023, 15(6), 398; https://doi.org/10.3390/toxins15060398 - 15 Jun 2023

Cited by 1 | Viewed by 2314

Abstract

Envenomations induced by animal bites and stings constitute a significant public health burden. Even though a standardized protocol does not exist, parenterally administered polyclonal antivenoms remain the mainstay in snakebite therapy. There is a prevailing opinion that their application by the i.m. route [...] Read more.

Envenomations induced by animal bites and stings constitute a significant public health burden. Even though a standardized protocol does not exist, parenterally administered polyclonal antivenoms remain the mainstay in snakebite therapy. There is a prevailing opinion that their application by the i.m. route has poor efficacy and that i.v. administration should preferentially be chosen in order to achieve better accomplishment of the antivenom therapeutic activity. Recently, it has been demonstrated that neutralization not only in the systemic circulation but also in the lymphatic system might be of great importance for the clinical outcome since it represents another relevant body compartment through which the absorption of the venom components occurs. In this review, the present-day and summarized knowledge of the laboratory and clinical findings on the i.v. and i.m. routes of antivenom administration is provided, with a special emphasis on the contribution of the lymphatic system to the process of venom elimination. Until now, antivenom-mediated neutralization has not yet been discussed in the context of the synergistic action of both blood and lymph. A current viewpoint might help to improve the comprehension of the venom/antivenom pharmacokinetics and the optimal approach for drug application. There is a great need for additional dependable, practical, well-designed studies, as well as more practice-related experience reports. As a result, opportunities for resolving long-standing disputes over choosing one therapeutic principle over another might be created, improving the safety and effectiveness of snakebite management. Full article

(This article belongs to the Special Issue Passive Immunotherapy and Its Applications in Envenomation and Infectious Diseases: In Tribute to Gaston Ramon (1886–1963))

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23 pages, 6122 KiB

Open AccessArticle

High-Resolution Magic Angle Spinning (HRMAS) NMR Identifies Oxidative Stress and Impairment of Energy Metabolism by Zearalenone in Embryonic Stages of Zebrafish (Danio rerio), Olive Flounder (Paralichthys olivaceus) and Yellowtail Snapper (Ocyurus chrysurus)

by Mark Annunziato, Narmin Bashirova, Muhamed N. H. Eeza, Ariel Lawson, Daniel Benetti, John D. Stieglitz, Jörg Matysik, A. Alia and John P. Berry

Toxins 2023, 15(6), 397; https://doi.org/10.3390/toxins15060397 - 15 Jun 2023

Cited by 1 | Viewed by 1314

Abstract

Zearalenone (ZEA) is a mycotoxin, commonly found in agricultural products, linked to adverse health impacts in humans and livestock. However, less is known regarding effects on fish as both ecological receptors and economically relevant “receptors” through contamination of aquaculture feeds. In the present [...] Read more.

Zearalenone (ZEA) is a mycotoxin, commonly found in agricultural products, linked to adverse health impacts in humans and livestock. However, less is known regarding effects on fish as both ecological receptors and economically relevant “receptors” through contamination of aquaculture feeds. In the present study, a metabolomics approach utilizing high-resolution magic angle spinning nuclear magnetic resonance (HRMAS NMR) was applied to intact embryos of zebrafish (Danio rerio), and two marine fish species, olive flounder (Paralichthys olivaceus) and yellowtail snapper (Ocyurus chrysurus), to investigate the biochemical pathways altered by ZEA exposure. Following the assessment of embryotoxicity, metabolic profiling of embryos exposed to sub-lethal concentrations showed significant overlap between the three species and, specifically, identified metabolites linked to hepatocytes, oxidative stress, membrane disruption, mitochondrial dysfunction, and impaired energy metabolism. These findings were further supported by analyses of tissue-specific production of reactive oxygen species (ROS) and lipidomics profiling and enabled an integrated model of ZEA toxicity in the early life stages of marine and freshwater fish species. The metabolic pathways and targets identified may, furthermore, serve as potential biomarkers for monitoring ZEA exposure and effects in fish in relation to ecotoxicology and aquaculture. Full article

(This article belongs to the Section Mycotoxins)

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8 pages, 1118 KiB

Open AccessCommunication

Is Proteolytic Cleavage Essential for the Enhanced Activity of Hydra Pore-Forming Toxin, HALT-4?

by Wei Yuen Yap and Jung Shan Hwang

Toxins 2023, 15(6), 396; https://doi.org/10.3390/toxins15060396 - 13 Jun 2023

Viewed by 830

Abstract

Hydra actinoporin-like toxin 4 (HALT-4) differs from other actinoporins due to its N-terminal propart that contains approximately 103 additional residues. Within this region, we identified five dibasic residues and assumed that, when cleaved, they could potentially exhibit HALT-4′s cytolytic activity. We created five [...] Read more.

Hydra actinoporin-like toxin 4 (HALT-4) differs from other actinoporins due to its N-terminal propart that contains approximately 103 additional residues. Within this region, we identified five dibasic residues and assumed that, when cleaved, they could potentially exhibit HALT-4′s cytolytic activity. We created five truncated versions of HALT-4 (tKK1, tKK2, tRK3, tKK4 and tKK5) to investigate the role of the N-terminal region and potential cleavage sites on the cytolytic activity of HALT-4. However, our results demonstrated that the propart-containing HALT-4 (proHALT-4), as well as the truncated versions tKK1 and tKK2, exhibited similar cytolytic activity against HeLa cells. In contrast, tRK3, tKK4 and tKK5 failed to kill HeLa cells, indicating that cleavage at the KK1 or KK2 sites did not enhance cytolytic activity but may instead facilitate the sorting of tKK1 and tKK2 to the regulated secretory pathway for eventual deposition in nematocysts. Moreover, RK3, KK4 and KK5 were unlikely to serve as proteolytic cleavage sites, as the amino acids between KK2 and RK3 are also crucial for pore formation. Full article

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

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19 pages, 3856 KiB

Open AccessArticle

Dissolved Algal Toxins along the Southern Coast of British Columbia Canada

by Ryan B. Shartau, Lenora D. M. Turcotte, Julia C. Bradshaw, Andrew R. S. Ross, Blair D. Surridge, Nina Nemcek and Stewart C. Johnson

Toxins 2023, 15(6), 395; https://doi.org/10.3390/toxins15060395 - 13 Jun 2023

Cited by 1 | Viewed by 1750

Abstract

Harmful algal blooms (HABs) in coastal British Columbia (BC), Canada, negatively impact the salmon aquaculture industry. One disease of interest to salmon aquaculture is Net Pen Liver Disease (NPLD), which induces severe liver damage and is believed to be caused by the exposure [...] Read more.

Harmful algal blooms (HABs) in coastal British Columbia (BC), Canada, negatively impact the salmon aquaculture industry. One disease of interest to salmon aquaculture is Net Pen Liver Disease (NPLD), which induces severe liver damage and is believed to be caused by the exposure to microcystins (MCs). To address the lack of information about algal toxins in BC marine environments and the risk they pose, this study investigated the presence of MCs and other toxins at aquaculture sites. Sampling was carried out using discrete water samples and Solid Phase Adsorption Toxin Tracking (SPATT) samplers from 2017–2019. All 283 SPATT samples and all 81 water samples tested positive for MCs. Testing for okadaic acid (OA) and domoic acid (DA) occurred in 66 and 43 samples, respectively, and all samples were positive for the toxin tested. Testing for dinophysistoxin-1 (DTX-1) (20 samples), pectenotoxin-2 (PTX-2) (20 samples), and yessotoxin (YTX) (17 samples) revealed that all samples were positive for the tested toxins. This study revealed the presence of multiple co-occurring toxins in BC’s coastal waters and the levels detected in this study were below the regulatory limits for health and recreational use. This study expands our limited knowledge of algal toxins in coastal BC and shows that further studies are needed to understand the risks they pose to marine fisheries and ecosystems. Full article

(This article belongs to the Special Issue Harmful Algal Blooms and Microalgae from Ecology to Control Strategies)

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17 pages, 4562 KiB

Open AccessArticle

Vitamin D Supplementation Impacts Calcium and Phosphorus Metabolism in Piglets Fed a Diet Contaminated with Deoxynivalenol and Challenged with Lipopolysaccharides

by Béatrice Sauvé, Younes Chorfi, Marie-Pierre Létourneau Montminy and Frédéric Guay

Toxins 2023, 15(6), 394; https://doi.org/10.3390/toxins15060394 - 13 Jun 2023

Cited by 2 | Viewed by 1548

Abstract

Using alternative feed ingredients in pig diets can lead to deoxynivalenol (DON) contamination. DON has been shown to induce anorexia, inflammation, and—more recently—alterations in the vitamin D, calcium, and phosphorus metabolisms. Adding vitamin D supplementation in the form of vitamin D₃ and [...] Read more.

Using alternative feed ingredients in pig diets can lead to deoxynivalenol (DON) contamination. DON has been shown to induce anorexia, inflammation, and—more recently—alterations in the vitamin D, calcium, and phosphorus metabolisms. Adding vitamin D supplementation in the form of vitamin D₃ and 25-OH-D₃ to the feed could modify the effects of DON in piglets. In this study, vitamin D₃ or 25-OH-D₃ supplementation was used in a control or DON-contaminated treatment. A repetitive exposure over 21 days to DON in the piglets led to disruptions in the vitamin D, calcium, and phosphorus metabolisms, resulting in a decreased growth performance, increased bone mineralization, and the downregulation of genes related to calcium and to phosphorus intestinal and renal absorption. The DON challenge also decreased blood concentrations of 25-OH-D₃, 1,25-(OH)₂-D₃, and phosphate. The DON contamination likely decreased the piglets’ vitamin D status indirectly by modifying the calcium metabolism response. Vitamin D supplementations did not restore vitamin D status or bone mineralization. After a lipopolysaccharide-induced inflammatory stimulation, feeding a 25-OH-D₃ supplementation increased 25-OH-D₃ concentration and 1,25-(OH)₂-D₃ regulations during the DON challenge. DON contamination likely induced a Ca afflux by altering the intestinal barrier, which resulted in hypercalcemia and hypovitaminosis D. The vitamin D supplementation could increase the calcitriol production to face the combined LPS and DON challenge. Full article

(This article belongs to the Special Issue Effects of Mycotoxins on Animals)

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13 pages, 8752 KiB

Open AccessArticle

Development of an Online Genome Sequence Comparison Resource for Bacillus cereus sensu lato Strains Using the Efficient Composition Vector Method

by Kui Wang, Changlong Shu, Alejandra Bravo, Mario Soberón, Hongjun Zhang, Neil Crickmore and Jie Zhang

Toxins 2023, 15(6), 393; https://doi.org/10.3390/toxins15060393 - 12 Jun 2023

Cited by 3 | Viewed by 1658

Abstract

An automated method was developed for differentiating closely related B. cereus sensu lato (s.l.) species, especially biopesticide Bacillus thuringiensis, from other human pathogens, B. anthracis and B. cereus sensu stricto (s.s.). In the current research, four typing methods were [...] Read more.

An automated method was developed for differentiating closely related B. cereus sensu lato (s.l.) species, especially biopesticide Bacillus thuringiensis, from other human pathogens, B. anthracis and B. cereus sensu stricto (s.s.). In the current research, four typing methods were initially compared, including multi-locus sequence typing (MLST), single-copy core genes phylogenetic analysis (SCCGPA), dispensable genes content pattern analysis (DGCPA) and composition vector tree (CVTree), to analyze the genomic variability of 23 B. thuringiensis strains from aizawai, kurstaki, israelensis, thuringiensis and morrisoni serovars. The CVTree method was the best option to be used for typing B. thuringiensis strains since it proved to be the fastest method, whilst giving high-resolution data about the strains. In addition, CVTree agrees well with ANI-based method, revealing the relationship between B. thuringiensis and other B. cereus s.l. species. Based on these data, an online genome sequence comparison resource was built for Bacillus strains called the Bacillus Typing Bioinformatics Database to facilitate strain identification and characterization. Full article

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15 pages, 10298 KiB

Open AccessArticle

Zearalenone Exposure Disrupts STAT-ISG15 in Rat Colon: A Potential Linkage between Zearalenone and Inflammatory Bowel Disease

by Haonan Ruan, Jiashuo Wu, Fangqing Zhang, Ziyue Jin, Jiao Tian, Jing Xia, Jiaoyang Luo and Meihua Yang

Toxins 2023, 15(6), 392; https://doi.org/10.3390/toxins15060392 - 09 Jun 2023

Viewed by 1395

Abstract

Zearalenone (ZEN), a prevalent mycotoxin contaminating food and known for its intestinal toxicity, has been suggested as a potential risk factor for inflammatory bowel disease (IBD), although the exact relationship between ZEN exposure and IBD remains unclear. In this study, we established a [...] Read more.

Zearalenone (ZEN), a prevalent mycotoxin contaminating food and known for its intestinal toxicity, has been suggested as a potential risk factor for inflammatory bowel disease (IBD), although the exact relationship between ZEN exposure and IBD remains unclear. In this study, we established a rat model of colon toxicity induced by ZEN exposure to investigate the key targets of ZEN-induced colon toxicity and explore the underlying connection between ZEN exposure and IBD. Histological staining of the rat colon revealed significant pathological changes resulting from ZEN exposure (p < 0.01). Furthermore, the proteomic analysis demonstrated a notable upregulation of protein expression levels, specifically STAT2 (0.12 ± 0.0186), STAT6 (0.36 ± 0.0475) and ISG15 (0.43 ± 0.0226) in the rat colon (p < 0.05). Utilizing bioinformatics analysis, we combined ZEN exposure and IBD clinical sample databases to reveal that ZEN exposure may increase the risk of IBD through activation of the STAT-ISG15 pathway. This study identified novel targets for ZEN-induced intestinal toxicity, providing the basis for further study of ZEN exposure to IBD. Full article

(This article belongs to the Special Issue Omics Approaches to Study Toxins)

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16 pages, 624 KiB

Open AccessReview

Improving the Efficacy of Botulinum Toxin for Cervical Dystonia: A Scoping Review

by Roberto Erro, Marina Picillo, Maria Teresa Pellecchia and Paolo Barone

Toxins 2023, 15(6), 391; https://doi.org/10.3390/toxins15060391 - 09 Jun 2023

Cited by 3 | Viewed by 3077

Abstract

Cervical dstonia (CD) is a chronic disorder with a significant detrimental impact on quality of life, requiring long-term treatment. Intramuscular injections of botulinum neurotoxin (BoNT) every 12 to 16 weeks have become the first-line option for CD. Despite the remarkable efficacy of BoNT [...] Read more.

Cervical dstonia (CD) is a chronic disorder with a significant detrimental impact on quality of life, requiring long-term treatment. Intramuscular injections of botulinum neurotoxin (BoNT) every 12 to 16 weeks have become the first-line option for CD. Despite the remarkable efficacy of BoNT as a treatment for CD, a significantly high proportion of patients report poor outcomes and discontinue the treatment. The reasons that drive sub-optimal response or treatment failure in a proportion of patients include but are not limited to inappropriate muscle targets and/or BoNT dosing, improper method of injections, subjective feeling of inefficacy, and the formation of neutralizing antibodies against the neurotoxin. The current review aims to complement published research focusing on the identification of the factors that might explain the failure of BoNT treatment in CD, highlighting possible solutions to improve its outcomes. Thus, the use of the new phenomenological classification of cervical dystonia known as COL-CAP might improve the identification of the muscle targets, but more sensitive information might come from the use of kinematic or scintigraphic techniques and the use of electromyographic or ultrasound guidance might ensure the accuracy of the injections. Suggestions are made for the development of a patient-centered model for the management of cervical dystonia and to emphasize that unmet needs in the field are to increase awareness about the non-motor spectrum of CD, which might influence the perception of the efficacy from BoNT injections, and the development of dedicated rehabilitation programs for CD that might enhance its effectiveness. Full article

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23 pages, 5583 KiB

Open AccessEditor’s ChoiceArticle

The Clostridium botulinum C2 Toxin Subunit C2IIa Delivers Enzymes with Positively Charged N-Termini into the Cytosol of Target Cells

by Sebastian Heber, Joscha Borho, Nicole Stadler, Fanny Wondany, Irina König, Jens Michaelis, Panagiotis Papatheodorou, Holger Barth and Maximilian Fellermann

Toxins 2023, 15(6), 390; https://doi.org/10.3390/toxins15060390 - 09 Jun 2023

Viewed by 1743

Abstract

The binary Clostridium (C.) botulinum C2 toxin consists of two non-linked proteins. The proteolytically activated binding/transport subunit C2IIa forms barrel-shaped homoheptamers, which bind to cell surface receptors, mediate endocytosis, and translocate the enzyme subunit C2I into the cytosol of target cells. [...] Read more.

The binary Clostridium (C.) botulinum C2 toxin consists of two non-linked proteins. The proteolytically activated binding/transport subunit C2IIa forms barrel-shaped homoheptamers, which bind to cell surface receptors, mediate endocytosis, and translocate the enzyme subunit C2I into the cytosol of target cells. Here, we investigate whether C2IIa can be harnessed as a transporter for proteins/enzymes fused to polycationic tags, as earlier demonstrated for the related anthrax toxin transport subunit PA₆₃. To test C2IIa-mediated transport in cultured cells, reporter enzymes are generated by fusing different polycationic tags to the N- or C-terminus of other bacterial toxins’ catalytic A subunits. C2IIa as well as PA₆₃ deliver N-terminally polyhistidine-tagged proteins more efficiently compared to C-terminally tagged ones. However, in contrast to PA₆₃, C2IIa does not efficiently deliver polylysine-tagged proteins into the cytosol of target cells. Moreover, untagged enzymes with a native cationic N-terminus are efficiently transported by both C2IIa and PA₆₃. In conclusion, the C2IIa-transporter serves as a transport system for enzymes that harbor positively charged amino acids at their N-terminus. The charge distribution at the N-terminus of cargo proteins and their ability to unfold in the endosome and subsequently refold in the cytosol determine transport feasibility and efficiency. Full article

(This article belongs to the Special Issue Intracellular Transport of Toxins: Insights into Mechanisms of Cytotoxicity and Applications in Cell Biology and Medicine)

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19 pages, 9115 KiB

Open AccessArticle

Natural Occurrence of Regulated and Emerging Mycotoxins in Wheat Grains and Assessment of the Risks from Dietary Mycotoxins Exposure in China

by Xiaofeng Ji, Canghong Jin, Yingping Xiao, Meihua Deng, Wen Wang, Wentao Lyu, Jiapeng Chen, Rui Li, Yan Li and Hua Yang

Toxins 2023, 15(6), 389; https://doi.org/10.3390/toxins15060389 - 09 Jun 2023

Cited by 4 | Viewed by 1544

Abstract

Wheat grains are susceptible to contamination with various natural mycotoxins including regulated and emerging mycotoxins. This study surveyed the natural presence of regulated mycotoxins, such as deoxynivalenol (DON) and zearalenone (ZEN), and emerging mycotoxins such as beauvericin (BEA), enniatins (ENNs such as ENA, [...] Read more.

Wheat grains are susceptible to contamination with various natural mycotoxins including regulated and emerging mycotoxins. This study surveyed the natural presence of regulated mycotoxins, such as deoxynivalenol (DON) and zearalenone (ZEN), and emerging mycotoxins such as beauvericin (BEA), enniatins (ENNs such as ENA, ENA1, ENB, ENB1) and Alternaria mycotoxins (i.e., alternariol monomethyl ether (AME), alternariol (AOH), tenuazonic acid (TeA), tentoxin (TEN), and altenuene (ALT)) in wheat grains randomly collected from eight provinces across China in 2021. The results revealed that each wheat grain sample was detected with at least one type of mycotoxin. The detection rates of these mycotoxins ranged from 7.1% to 100%, with the average occurrence level ranging from 1.11 to 921.8 µg/kg. DON and TeA were the predominant mycotoxins with respect to both prevalence and concentration. Approximately 99.7% of samples were found to contain more than one toxin, and the co-occurrence of ten toxins (DON + ZEN + ENA + ENA1 + ENB + ENB1 + AME + AOH + TeA + TEN) was the most frequently detected combination. The dietary exposure to different mycotoxins among Chinese consumers aged 4–70 years was as follows: 0.592–0.992 µg/kg b.w./day for DON, 0.007–0.012 µg/kg b.w./day for ZEN, 0.0003–0.007 µg/kg b.w./day for BEA and ENNs, 0.223–0.373 µg/kg b.w./day for TeA, and 0.025–0.041 µg/kg b.w./day for TEN, which were lower than the health-based guidance values for each mycotoxin, with the corresponding hazard quotient (HQ) being far lower than 1, implying a tolerable health risk for Chinese consumers. However, the estimated dietary exposure to AME and AOH was in the range of 0.003–0.007 µg/kg b.w./day, exceeding the Threshold of Toxicological Concern (TTC) value of 0.0025 µg/kg b.w./day, demonstrating potential dietary risks for Chinese consumers. Therefore, developing practical control and management strategies is essential for controlling mycotoxins contamination in the agricultural systems, thereby ensuring public health. Full article

(This article belongs to the Section Mycotoxins)

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13 pages, 1097 KiB

Open AccessReview

Cyanotoxins and Other Bioactive Compounds from the Pasteur Cultures of Cyanobacteria (PCC)

by Muriel Gugger, Anne Boullié and Thierry Laurent

Toxins 2023, 15(6), 388; https://doi.org/10.3390/toxins15060388 - 09 Jun 2023

Cited by 2 | Viewed by 1668

Abstract

In tribute to the bicentenary of the birth of Louis Pasteur, this report focuses on cyanotoxins, other natural products and bioactive compounds of cyanobacteria, a phylum of Gram-negative bacteria capable of carrying out oxygenic photosynthesis. These microbes have contributed to changes in the [...] Read more.

In tribute to the bicentenary of the birth of Louis Pasteur, this report focuses on cyanotoxins, other natural products and bioactive compounds of cyanobacteria, a phylum of Gram-negative bacteria capable of carrying out oxygenic photosynthesis. These microbes have contributed to changes in the geochemistry and the biology of Earth as we know it today. Furthermore, some bloom-forming cyanobacterial species are also well known for their capacity to produce cyanotoxins. This phylum is preserved in live cultures of pure, monoclonal strains in the Pasteur Cultures of Cyanobacteria (PCC) collection. The collection has been used to classify organisms within the Cyanobacteria of the bacterial kingdom and to investigate several characteristics of these bacteria, such as their ultrastructure, gas vacuoles and complementary chromatic adaptation. Thanks to the ease of obtaining genetic and further genomic sequences, the diversity of the PCC strains has made it possible to reveal some main cyanotoxins and to highlight several genetic loci dedicated to completely unknown natural products. It is the multidisciplinary collaboration of microbiologists, biochemists and chemists and the use of the pure strains of this collection that has allowed the study of several biosynthetic pathways from genetic origins to the structures of natural products and, eventually, their bioactivity. Full article

(This article belongs to the Special Issue Pasteur, Toxins/Pathogenicity, Anti-toxins, a Bicentennial Contribution)

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15 pages, 2901 KiB

Open AccessArticle

Induced Expression of the Acinetobacter sp. Oxa Gene in Lactobacillus acidophilus and Its Increased ZEN Degradation Stability by Immobilization

by Yuqun Zhou, An Wang, Qingzi Yu, Yuqian Tang and Yuanshan Yu

Toxins 2023, 15(6), 387; https://doi.org/10.3390/toxins15060387 - 08 Jun 2023

Cited by 2 | Viewed by 1229

Abstract

Zearalenone (ZEN, ZEA) contamination in various foods and feeds is a significant global problem. Similar to deoxynivalenol (DON) and other mycotoxins, ZEN in feed mainly enters the body of animals through absorption in the small intestine, resulting in estrogen-like toxicity. In this study, [...] Read more.

Zearalenone (ZEN, ZEA) contamination in various foods and feeds is a significant global problem. Similar to deoxynivalenol (DON) and other mycotoxins, ZEN in feed mainly enters the body of animals through absorption in the small intestine, resulting in estrogen-like toxicity. In this study, the gene encoding Oxa, a ZEN-degrading enzyme isolated from Acinetobacter SM04, was cloned into Lactobacillus acidophilus ATCC4356, a parthenogenic anaerobic gut probiotic, and the 38 kDa sized Oxa protein was expressed to detoxify ZEN intestinally. The transformed strain L. acidophilus pMG-Oxa acquired the capacity to degrade ZEN, with a degradation rate of 42.95% at 12 h (initial amount: 20 μg/mL). The probiotic properties of L. acidophilus pMG-Oxa (e.g., acid tolerance, bile salt tolerance, and adhesion properties) were not affected by the insertion and intracellular expression of Oxa. Considering the low amount of Oxa expressed by L. acidophilus pMG-Oxa and the damage to enzyme activity by digestive juices, Oxa was immobilized with 3.5% sodium alginate, 3.0% chitosan, and 0.2 M CaCl₂ to improve the ZEN degradation efficiency (from 42.95% to 48.65%) and protect it from digestive juices. The activity of immobilized Oxa was 32–41% higher than that of the free crude enzyme at different temperatures (20–80 °C), pH values (2.0–12.0), storage conditions (4 °C and 25 °C), and gastrointestinal simulated digestion conditions. Accordingly, immobilized Oxa could be resistant to adverse environmental conditions. Owing to the colonization, efficient degradation performance, and probiotic functionality of L. acidophilus, it is an ideal host for detoxifying residual ZEN in vivo, demonstrating great potential for application in the feed industry. Full article

(This article belongs to the Special Issue Mitigation and Detoxification Strategies of Mycotoxins)

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9 pages, 933 KiB

Open AccessCommunication

Transgenic Drosophila to Functionally Validate Fall Armyworm ABCC2 Mutations Conferring Bt Resistance

by Rafaela Panteleri, Amalia Anthousi, Shane Denecke, Debora Boaventura, Ralf Nauen and John Vontas

Toxins 2023, 15(6), 386; https://doi.org/10.3390/toxins15060386 - 07 Jun 2023

Viewed by 1991

Abstract

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) is an invasive agricultural pest with a global distribution, causing major crop losses annually. Its control strategies largely rely on chemical insecticides and transgenic crops expressing Bacillus thuringiensis insecticidal proteins (Cry and Vip [...] Read more.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) is an invasive agricultural pest with a global distribution, causing major crop losses annually. Its control strategies largely rely on chemical insecticides and transgenic crops expressing Bacillus thuringiensis insecticidal proteins (Cry and Vip toxins); however, the development of high resistance poses a significant issue. The ATP-binding cassette transporter C2 (ABCC2) has been linked to Cry toxin pore formation, acting as a receptor of some Cry toxins. Recently detected mutations in the SfABCC2 gene in extracellular loop 4 (ECL4) have been associated with Bt toxin resistance in FAW. In the present study, we expressed the SfABCC2 gene in Drosophila melanogaster, a species normally unaffected by the Bt toxins. We demonstrate that susceptibility can be introduced by the ectopic and tissue-specific expression of wildtype SfABCC2. Next, we introduced mutations into ECL4—both individually and in combination—that have been recently described in Brazilian FAW and functionally validated by toxicity bioassays against the foliar Bt product Xentari. Our results provide an efficient demonstration of the suitability of transgenic Drosophila for validating FAW ABCC2 resistance mutations in ECL4 against Bt toxins, and potential cross-resistance issues between closely related proteins that use ABCC2. Full article

(This article belongs to the Special Issue Resistance to Insect-Killing Proteins from the Bacillus thuringiensis (Bt) Toxins)

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12 pages, 916 KiB

Open AccessArticle

Botulinum Toxin Therapy for Psychiatric Disorders in Clinical Practice: A Retrospective Case Study

by Franziska Lehnert, Insa Neumann, Tillmann H. C. Krüger and Marc A. Wollmer

Toxins 2023, 15(6), 385; https://doi.org/10.3390/toxins15060385 - 07 Jun 2023

Cited by 1 | Viewed by 1528

Abstract

Inhibiting the facial expression of negative emotions via botulinum toxin A (BTX) has been shown to mitigate symptoms of clinical depression in randomized controlled trials. This retrospective case study sought to reproduce the beneficial effects of BTX in a naturalistic setting for major [...] Read more.

Inhibiting the facial expression of negative emotions via botulinum toxin A (BTX) has been shown to mitigate symptoms of clinical depression in randomized controlled trials. This retrospective case study sought to reproduce the beneficial effects of BTX in a naturalistic setting for major depressive disorder and collect casuistic data on its effect on other mental disorders. Moreover, we describe symptom development across multiple treatment cycles with BTX, and assess the implementation of additional injection targets in the lower face region. Participants were N = 51 adult psychiatric outpatients mainly seeking treatment for depression. Over 50% suffered from comorbid psychiatric conditions, predominantly generalized anxiety disorder (GAD) or borderline personality disorder (BPD). A pre–post case series design was adapted. All participants received BTX-injections in the glabellar region on at least one occasion. Some received additional injections in the mouth region and over multiple treatment cycles. Treatment response was followed up by self-rated scales at varying time intervals post treatment. The results showed that BTX may yield favorable outcomes across multiple and comorbid mental disorders, especially, however, for patients suffering from depression. It potentially prevents the recurrence of clinical symptoms if applied regularly. Adding additional regions of the face does not seem to be superior over applying it to the glabellar region alone. The results add to the growing evidence that BTX therapy is effective in alleviating symptoms of depression. Positive effects can be sustained and reinstated, when applied over multiple treatment cycles. Observed symptom reduction in other psychiatric disorders was less pronounced. Further research is needed to understand the mechanisms by which BTX therapy reduces psychiatric symptoms. Full article

(This article belongs to the Special Issue Treatment of Mental Health Disorders with Botulinum Toxin)

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15 pages, 7662 KiB

Open AccessFeature PaperEditor’s ChoiceArticle

Domperidone Protects Cells from Intoxication with Clostridioides difficile Toxins by Inhibiting Hsp70-Assisted Membrane Translocation

by Maria Braune-Yan, Jinfang Jia, Mary Wahba, Johannes Schmid, Panagiotis Papatheodorou, Holger Barth and Katharina Ernst

Toxins 2023, 15(6), 384; https://doi.org/10.3390/toxins15060384 - 07 Jun 2023

Cited by 1 | Viewed by 1627

Abstract

Clostridioides difficile infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into [...] Read more.

Clostridioides difficile infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into the cytosol. The enzyme domains glucosylate small GTPases such as Rac1, thereby inhibiting processes such as actin cytoskeleton regulation. Here, we demonstrate that specific pharmacological inhibition of Hsp70 activity protected cells from TcdB intoxication. In particular, the established inhibitor VER-155008 and the antiemetic drug domperidone, which was found to be an Hsp70 inhibitor, reduced the number of cells with TcdB-induced intoxication morphology in HeLa, Vero and intestinal CaCo-2 cells. These drugs also decreased the intracellular glucosylation of Rac1 by TcdB. Domperidone did not inhibit TcdB binding to cells or enzymatic activity but did prevent membrane translocation of TcdB’s glucosyltransferase domain into the cytosol. Domperidone also protected cells from intoxication with TcdA as well as CDT toxin produced by hypervirulent strains of Clostridioides difficile. Our results reveal Hsp70 requirement as a new aspect of the cellular uptake mechanism of TcdB and identified Hsp70 as a novel drug target for potential therapeutic strategies required to combat severe Clostridioides difficile infections. Full article

(This article belongs to the Special Issue Toxin-Host Interaction of Clostridium Toxins)

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13 pages, 1061 KiB

Open AccessReview

Enniatin B1: Emerging Mycotoxin and Emerging Issues

by Beatrice De Felice, Leon J. Spicer and Francesca Caloni

Toxins 2023, 15(6), 383; https://doi.org/10.3390/toxins15060383 - 06 Jun 2023

Cited by 4 | Viewed by 1732

Abstract

Although over the last 10 years several studies have focused on the emerging mycotoxins known as enniatins (ENNs), there is still a lack of knowledge regarding their toxicological effects and the development of a correct risk assessment. This is especially true for enniatin [...] Read more.

Although over the last 10 years several studies have focused on the emerging mycotoxins known as enniatins (ENNs), there is still a lack of knowledge regarding their toxicological effects and the development of a correct risk assessment. This is especially true for enniatin B1 (ENN B1), considered the younger sister of the widely studied enniatin B (ENN B). ENN B1 has been found in several food commodities and, as with other mycotoxins, presents antibacterial and antifungal properties. On the other hand, ENN B1 has shown cytotoxic activity, impairment of the cell cycle, the induction of oxidative stress, and changes in mitochondrial membrane permeabilization, as well as negative genotoxic and estrogenic effects. Overall, considering the paucity of information available regarding ENN B1, further studies are necessary to perform a risk assessment. This review summarizes information on the biological characteristics and toxicological effects of ENN B1 as well as the future challenges that this mycotoxin could present. Full article

(This article belongs to the Collection Editorial Board Members’ Collection Series: Fungal Metabolites: From Toxins to Therapeutics)

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9 pages, 1215 KiB

Open AccessCommunication

Safety and Effectiveness of Repeated Botulinum Toxin A Intracavernosal Injections in Men with Erectile Dysfunction Unresponsive to Approved Pharmacological Treatments: Real-World Observational Data

by François Giuliano, Pierre Denys and Charles Joussain

Toxins 2023, 15(6), 382; https://doi.org/10.3390/toxins15060382 - 05 Jun 2023

Viewed by 2710

Abstract

Intracavernosal injections of botulinum toxin A (BTX/A ic) may be effective for difficult-to-treat erectile dysfunction (ED). This is a retrospective case series study of the effectiveness of repeated off-label BTX/A ic (onabotulinumtoxinA 100U, incobotulinumtoxinA 100U or abobotulinumtoxinA 500U) in men with ED [...] Read more.

Intracavernosal injections of botulinum toxin A (BTX/A ic) may be effective for difficult-to-treat erectile dysfunction (ED). This is a retrospective case series study of the effectiveness of repeated off-label BTX/A ic (onabotulinumtoxinA 100U, incobotulinumtoxinA 100U or abobotulinumtoxinA 500U) in men with ED and insufficient response to phosphodiesterase type 5 inhibitors (PDE5-Is) or prostaglandinE1 intracavernosal injections (PGE1 ICIs), defined as an International Index of Erectile Function-Erectile Function domain score (IIEF-EF) < 26 on treatment. Further injections were performed on patients’ requests, and the files of men who underwent at least two injections were reviewed. The response to BTX/A ic was defined as the achievement of the minimally clinically important difference in IIEF-EF adjusted to the severity of ED on treatment at baseline. Out of 216 men treated with BTX/A ic and PDE5-Is or PGE1-ICIs, 92 (42.6%) requested at least a second injection. The median time since the preceding injection was 8.7 months. In total, 85, 44 and 23 men received, respectively, two, three and four BTX/A ic. The overall response rate was 77.5%: 85.7% in men with mild ED, 79% for moderate ED and 64.3% for severe ED on treatment. The response increased with repeated injections: 67.5%, 87.5% and 94.7%, respectively, after the second, third and fourth injections. Post-injection changes in IIEF-EF were similar across injections. The time from injection to request for a further injection varied little. Four men reported penile pain at the time of injection (1.5% of all injections), and one experienced a burn at the penile crus. Repeated BTX/A injections combined with PDE5-Is or PGE1-ICIs produced an effective and durable response, with acceptable safety. Full article

(This article belongs to the Special Issue Clinical Application of Botulinum Toxin 2.0)

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12 pages, 3197 KiB

Open AccessArticle

Screening Biocontrol Agents for Cash Crop Fusarium Wilt Based on Fusaric Acid Tolerance and Antagonistic Activity against Fusarium oxysporum

by Qinggang Guo, Shixin Li, Lihong Dong, Zhenhe Su, Peipei Wang, Xiaomeng Liu and Ping Ma

Toxins 2023, 15(6), 381; https://doi.org/10.3390/toxins15060381 - 05 Jun 2023

Cited by 1 | Viewed by 1534

Abstract

Fusarium wilt, caused by Fusarium oxysporum, is one of the most notorious diseases of cash crops. The use of microbial fungicides is an effective measure for controlling Fusarium wilt, and the genus Bacillus is an important resource for the development of microbial [...] Read more.

Fusarium wilt, caused by Fusarium oxysporum, is one of the most notorious diseases of cash crops. The use of microbial fungicides is an effective measure for controlling Fusarium wilt, and the genus Bacillus is an important resource for the development of microbial fungicides. Fusaric acid (FA) produced by F. oxysporum can inhibit the growth of Bacillus, thus affecting the control efficacy of microbial fungicides. Therefore, screening FA-tolerant biocontrol Bacillus may help to improve the biocontrol effect on Fusarium wilt. In this study, a method for screening biocontrol agents against Fusarium wilt was established based on tolerance to FA and antagonism against F. oxysporum. Three promising biocontrol bacteria, named B31, F68, and 30833, were obtained to successfully control tomato, watermelon, and cucumber Fusarium wilt. Strains B31, F68, and 30833 were identified as B. velezensis by phylogenetic analysis of the 16S rDNA, gyrB, rpoB, and rpoC gene sequences. Coculture assays revealed that strains B31, F68, and 30833 showed increased tolerance to F. oxysporum and its metabolites compared with B. velezensis strain FZB42. Further experiments confirmed that 10 µg/mL FA completely inhibited the growth of strain FZB42, while strains B31, F68, and 30833 maintained normal growth at 20 µg/mL FA and partial growth at 40 µg/mL FA. Compared with strain FZB42, strains B31, F68, and 30833 exhibited significantly greater tolerance to FA. Full article

(This article belongs to the Special Issue Detoxification, Control, and Toxicological Studies on Mycotoxins through Biotechnological Approaches)

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30 pages, 1287 KiB

Open AccessReview

Bacterial Toxin-Antitoxin Systems’ Cross-Interactions—Implications for Practical Use in Medicine and Biotechnology

by Lidia Boss and Barbara Kędzierska

Toxins 2023, 15(6), 380; https://doi.org/10.3390/toxins15060380 - 04 Jun 2023

Cited by 2 | Viewed by 2647

Abstract

Toxin-antitoxin (TA) systems are widely present in bacterial genomes. They consist of stable toxins and unstable antitoxins that are classified into distinct groups based on their structure and biological activity. TA systems are mostly related to mobile genetic elements and can be easily [...] Read more.

Toxin-antitoxin (TA) systems are widely present in bacterial genomes. They consist of stable toxins and unstable antitoxins that are classified into distinct groups based on their structure and biological activity. TA systems are mostly related to mobile genetic elements and can be easily acquired through horizontal gene transfer. The ubiquity of different homologous and non-homologous TA systems within a single bacterial genome raises questions about their potential cross-interactions. Unspecific cross-talk between toxins and antitoxins of non-cognate modules may unbalance the ratio of the interacting partners and cause an increase in the free toxin level, which can be deleterious to the cell. Moreover, TA systems can be involved in broadly understood molecular networks as transcriptional regulators of other genes’ expression or modulators of cellular mRNA stability. In nature, multiple copies of highly similar or identical TA systems are rather infrequent and probably represent a transition stage during evolution to complete insulation or decay of one of them. Nevertheless, several types of cross-interactions have been described in the literature to date. This implies a question of the possibility and consequences of the TA system cross-interactions, especially in the context of the practical application of the TA-based biotechnological and medical strategies, in which such TAs will be used outside their natural context, will be artificially introduced and induced in the new hosts. Thus, in this review, we discuss the prospective challenges of system cross-talks in the safety and effectiveness of TA system usage. Full article

(This article belongs to the Special Issue Intracellular Transport of Toxins: Insights into Mechanisms of Cytotoxicity and Applications in Cell Biology and Medicine)

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14 pages, 355 KiB

Open AccessReview

Mycotoxin Determination and Occurrence in Pseudo-Cereals Intended for Food and Feed: A Review

by María Vanessa Vila-López, Noelia Pallarés, Emilia Ferrer and Josefa Tolosa

Toxins 2023, 15(6), 379; https://doi.org/10.3390/toxins15060379 - 04 Jun 2023

Cited by 2 | Viewed by 1519

Abstract

Nowadays, pseudo-cereals’ consumption is increasing due to their health benefits as they possess an excellent nutrient profile. Whole pseudo-cereal grains are rich in a wide range of compounds, namely flavonoids, phenolic acids, fatty acids, and vitamins with known beneficial effects on human and [...] Read more.

Nowadays, pseudo-cereals’ consumption is increasing due to their health benefits as they possess an excellent nutrient profile. Whole pseudo-cereal grains are rich in a wide range of compounds, namely flavonoids, phenolic acids, fatty acids, and vitamins with known beneficial effects on human and animal health. Mycotoxins are common contaminants in cereals and by-products; however, the study of their natural occurrence in pseudo-cereals is currently scarce. Pseudo-cereals are similar to cereal grains; thus, mycotoxin contamination is expected to occur in pseudo-cereals. Indeed, mycotoxin-producing fungi have been reported in these matrices and, consequently, mycotoxin contents have been reported too, especially in buckwheat samples, where ochratoxin A and deoxynivalenol reached levels up to 1.79 μg/kg and 580 μg/kg, respectively. In comparison to cereal contamination, mycotoxin levels detected in pseudo-cereal samples are lower; however, more studies are necessary in order to describe the mycotoxin pattern in these samples and to establish maximum levels that ensure human and animal health protection. In this review, mycotoxin occurrence in pseudo-cereal samples as well as the main extraction methods and analytical techniques to determine them are described, showing that mycotoxins can be present in pseudo-cereal samples and that the most employed techniques for their determination are liquid and gas chromatography coupled to different detectors. Full article

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

24 pages, 8927 KiB

Open AccessFeature PaperArticle

Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Phα1β Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer

by Ekaterina N. Lyukmanova, Pavel A. Mironov, Dmitrii S. Kulbatskii, Mikhail A. Shulepko, Alexander S. Paramonov, Elizaveta M. Chernaya, Yulia A. Logashina, Yaroslav A. Andreev, Mikhail P. Kirpichnikov and Zakhar O. Shenkarev

Toxins 2023, 15(6), 378; https://doi.org/10.3390/toxins15060378 - 03 Jun 2023

Cited by 3 | Viewed by 1971

Abstract

Phα1β (PnTx3–6) is a neurotoxin from the spider Phoneutria nigriventer venom, originally identified as an antagonist of two ion channels involved in nociception: N-type voltage-gated calcium channel (Ca_V2.2) and TRPA1. In animal models, Phα1β administration reduces both acute and chronic pain. [...] Read more.

Phα1β (PnTx3–6) is a neurotoxin from the spider Phoneutria nigriventer venom, originally identified as an antagonist of two ion channels involved in nociception: N-type voltage-gated calcium channel (Ca_V2.2) and TRPA1. In animal models, Phα1β administration reduces both acute and chronic pain. Here, we report the efficient bacterial expression system for the recombinant production of Phα1β and its ¹⁵N-labeled analogue. Spatial structure and dynamics of Phα1β were determined via NMR spectroscopy. The N-terminal domain (Ala1–Ala40) contains the inhibitor cystine knot (ICK or knottin) motif, which is common to spider neurotoxins. The C-terminal α-helix (Asn41–Cys52) stapled to ICK by two disulfides exhibits the µs–ms time-scale fluctuations. The Phα1β structure with the disulfide bond patterns Cys1–5, Cys2–7, Cys3–12, Cys4–10, Cys6–11, Cys8–9 is the first spider knottin with six disulfide bridges in one ICK domain, and is a good reference to other toxins from the ctenitoxin family. Phα1β has a large hydrophobic region on its surface and demonstrates a moderate affinity for partially anionic lipid vesicles at low salt conditions. Surprisingly, 10 µM Phα1β significantly increases the amplitude of diclofenac-evoked currents and does not affect the allyl isothiocyanate (AITC)-evoked currents through the rat TRPA1 channel expressed in Xenopus oocytes. Targeting several unrelated ion channels, membrane binding, and the modulation of TRPA1 channel activity allow for considering Phα1β as a gating modifier toxin, probably interacting with S1–S4 gating domains from a membrane-bound state. Full article

(This article belongs to the Special Issue Ion Channels, Venom, and Toxins)

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22 pages, 5156 KiB

Open AccessArticle

Multi-Omic Identification of Venom Proteins Collected from Artificial Hosts of a Parasitoid Wasp

by Kaili Yu, Jin Chen, Xue Bai, Shijiao Xiong, Xinhai Ye, Yi Yang, Hongwei Yao, Fang Wang, Qi Fang, Qisheng Song and Gongyin Ye

Toxins 2023, 15(6), 377; https://doi.org/10.3390/toxins15060377 - 03 Jun 2023

Viewed by 1642

Abstract

Habrobracon hebetor is a parasitoid wasp capable of infesting many lepidopteran larvae. It uses venom proteins to immobilize host larvae and prevent host larval development, thus playing an important role in the biocontrol of lepidopteran pests. To identify and characterize its venom proteins, [...] Read more.

Habrobracon hebetor is a parasitoid wasp capable of infesting many lepidopteran larvae. It uses venom proteins to immobilize host larvae and prevent host larval development, thus playing an important role in the biocontrol of lepidopteran pests. To identify and characterize its venom proteins, we developed a novel venom collection method using an artificial host (ACV), i.e., encapsulated amino acid solution in paraffin membrane, allowing parasitoid wasps to inject venom. We performed protein full mass spectrometry analysis of putative venom proteins collected from ACV and venom reservoirs (VRs) (control). To verify the accuracy of proteomic data, we also collected venom glands (VGs), Dufour’s glands (DGs) and ovaries (OVs), and performed transcriptome analysis. In this paper, we identified 204 proteins in ACV via proteomic analysis; compared ACV putative venom proteins with those identified in VG, VR, and DG via proteome and transcriptome approaches; and verified a set of them using quantitative real-time polymerase chain reaction. Finally, 201 ACV proteins were identified as potential venom proteins. In addition, we screened 152 and 148 putative venom proteins identified in the VG transcriptome and the VR proteome against those in ACV, and found only 26 and 25 putative venom proteins, respectively, were overlapped with those in ACV. Altogether, our data suggest proteome analysis of ACV in combination with proteome–transcriptome analysis of other organs/tissues will provide the most comprehensive identification of true venom proteins in parasitoid wasps. Full article

(This article belongs to the Special Issue Animal Toxins: Biodiscovery, Mechanistic Insights and Translational Potential)

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10 pages, 599 KiB

Open AccessArticle

Outcomes of IncobotulinumtoxinA Injection on Myalgia and Arthralgia in Patients Undergoing Temporomandibular Joint Arthroscopy: A Randomized Controlled Trial

by David Faustino Ângelo, David Sanz, Francesco Maffia and Henrique José Cardoso

Toxins 2023, 15(6), 376; https://doi.org/10.3390/toxins15060376 - 03 Jun 2023

Cited by 4 | Viewed by 1431

Abstract

Background: Several studies have considered Botulinum Neurotoxin Type A injections effective in treating temporomandibular joint disorder (TMD) symptoms. A double-blind, randomized, controlled clinical trial investigated the benefit of complementary incobotulinumtoxinA (inco-BoNT/A) injections in the masticatory muscles of patients submitted to bilateral temporomandibular joint [...] Read more.

Background: Several studies have considered Botulinum Neurotoxin Type A injections effective in treating temporomandibular joint disorder (TMD) symptoms. A double-blind, randomized, controlled clinical trial investigated the benefit of complementary incobotulinumtoxinA (inco-BoNT/A) injections in the masticatory muscles of patients submitted to bilateral temporomandibular joint (TMJ) arthroscopy. Methods: Fifteen patients with TMD and an indication for bilateral TMJ arthroscopy were randomized into inco-BoNT/A (Xeomin, 100 U) or placebo groups (saline solution). Injections were carried out five days before TMJ arthroscopy. The primary outcome variable was a Visual Analogue Scale for TMJ arthralgia, and secondary outcomes were the myalgia degree, maximum mouth opening, and joint clicks. All outcome variables were assessed preoperatively (T0) and postoperatively (T1—week 5; T2—6-month follow-up). Results: At T1, the outcomes in the inco-BoNT/A group were improved, but not significantly more than in the placebo group. At T2, significant improvements in the TMJ arthralgia and myalgia scores were observed in the inco-BoNT/A group compared to the placebo. A higher number of postoperative reinterventions with further TMJ treatments were observed in the placebo group compared to inco-BoNT/A (63% vs. 14%). Conclusions: In patients submitted to TMJ arthroscopy, statistically significant long-term differences were observed between the placebo and inco-BoNT/A groups. Full article

(This article belongs to the Special Issue Botulinum Neurotoxin: Shared/Common Mechanisms in the Treatment of Pain, Spasticity and Movement Disorders)

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Toxins, Volume 15, Issue 6 (June 2023) – 55 articles

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