Amanita Phalloides Research Articles

S4420 The Shroom of Doom: Acute Liver Injury from Death Cap Mushroom Ingestion Treated With Experimental IV Silibinin

S4420 The Shroom of Doom: Acute Liver Injury from Death Cap Mushroom Ingestion Treated With Experimental IV Silibinin

Detection and quantification of Amatoxin in wild mushrooms from North-East India using HPLC-PDA method for food safety purposes

Misidentification and ingestion of poisonous mushrooms pose significant threats to food safety, particularly in Mizoram, India, where over ten fatalities due to mushroom poisoning have been reported in the past decade (2013-2023). This study aimed to address this critical issue by identifying and quantifying the cause of death due to consumption of wild mushroom from Champhai district, Mizoram, India and to test the reliability of HPLC-PDA for detection and quantification of amatoxins. HPLC-PDA confirmed the presence of α-amanitin in Amanita virosa and Amanita bisporigera in the samples. α-amanitin is a water-soluble, heat-stable, and highly toxic cyclic octapeptide present in the genus Amanita, which includes Amanita phalloides, Amanita verna, and Amanita virosa. Amanitin cytotoxicity arises from the inhibition of RNA polymerases, namely RNA polymerase II, which obstructs mRNA production in kidney and liver cells. Validation of the method demonstrated good precision and accuracy, with LOD and LOQ values of 88 ng g-1 and 210 ng g-1, respectively. The method was successfully applied to quantify α-amanitin in ten wild mushroom samples, revealing its presence only in Amanita virosa (1.17 mg g-1) and Amanita bisporigera (1.91 mg g-1) species. These findings underscore the importance of accurate α-amanitin detection methods in ensuring food safety and public health, particularly in regions prone to mushroom poisoning incidents. It is noteworthy that this study marks the initial exploration for detection and quantification of α-amanitin from poisonous mushrooms found in the wild regions of Champhai district in Mizoram, representing the first report of such in the area.

Establishment and validation of a nomogram model for prediction of clinical outcomes in patients with amanita phalloides poisoning

Amanita phalloides poisoning, renowned for its high mortality rates, is one of the most serious food safety issue in certain regions worldwide. Assessment of prognosis and development of more efficacious therapeutic strategies are critical importance for amanita phalloides poisoning patients. The aim of the study is to establish a nomogram to predict the clinical outcome of amanita phalloides poisoning patients based on the independent risk factor for prognosis. Herein, between January 2013 and September 2023, a cohort of 149 patients diagnosed with amanita phalloides poisoning was enrolled and randomly allocated into training and validation cohorts, comprising 102 and 47 patients, respectively. Multivariate logistic regression analysis was performed to identify the independent risk factors for morality of amanita phalloides poisoning patients in training cohort. Subsequently, a nomogram model was constructed to visually display the risk prediction model. The predictive accuracy of nomogram was verified by the validation cohort. The C index, the area under the receiver operating characteristic curve (AUC), and calibration plots were used to assessed the performance of nomogram. The clinical utility was evaluated by decision curve analysis (DCA). In the present study, the results showed that hepatic encephalopathy (HE), upper gastrointestinal bleeding (UGB), AST, and PT were the independent risk factors associated with the mortality of amantia phalloides poisoning patients. We constructed a new nomogram to evaluate the probability of death induced by amantia phalloides poisoning. The AUC for the prediction accuracy of the nomogram was 0.936 for the training cohort and 0.929 for the validation cohort. The calibration curves showed that the predicted probability matched the actual likelihood. The results of the DCA suggested that the nomogram has a good potential for clinical application. In summary, we developed a new nomogram to assess the probability of mortality for amanita phalloides poisoning patients. This nomogram might facilitate clinicians in making more efficacious treatment strategies for patients with amanita phalloides poisoning.

Chitosan extraction from Amanita phalloides: yield, crystallinity, degree of deacetylation, azo dye removal and antibacterial properties.

Chitosan, a biopolymer obtained from chitin, is known for its remarkable adsorption abilities for dyes, drugs, and fats, and its diverse array of antibacterial characteristics. This study explores the extraction and characterization of chitosan from the mycelium of Amanita phalloides. The moisture content, ash content, water binding capacity, fat binding capacity, and degree of deacetylation of the extracted chitosan were determined. The chitosan exhibited a high yield of 70%, crystallinity of 49.07%, a degree of deacetylation of 86%, and potent antimicrobial properties against both Gram-negative and Gram-positive bacteria. The study also examined the adsorption capabilities of chitosan to remove methylene blue (MB) dye by analysing specific factors like pH, reaction time, and MB concentration using the response surface model. The highest degree of MB dye removal was 91.6% at a pH of 6, a reaction time of around 60min and an initial dye concentration of 16ppm. This experimental design can be applied for chitosan adsorption of other organic compounds such as dyes, proteins, drugs, and fats.

Salidroside Inhibits α-Amanitin-Induced AML-12 Cell Apoptosis via the Regulation of PINK1/Parkin-Mediated Mitophagy and Mitochondrial Function.

Poisoning caused by the mushroom Amanita phalloides, due to the toxin α-amanitin, accounts for approximately 90% of food poisoning deaths in China with no specific antidotes. To investigate the role of salidroside (Sal) in α-amanitin (α-AMA)-induced mitophagy, mouse liver cells AML-12 were exposed to α-AMA in the presence of Sal or not. Intracellular reactive oxygen species (ROS) levels were measured using a ROS detection kit, mitochondrial activity was evaluated using a mitochondrial red fluorescent probe kit or JC-1 dye, and protein expression levels of PINK1, Parkin, LC3 II, P62, Bax, Bcl-2, Caspase 3, Cleaved-Caspase 3, PARP I, and Cleaved-PARP I were detected through Western blot. Results demonstrated that α-AMA led to increased intracellular ROS levels, cell apoptosis, and decreased mitochondrial membrane potential. Notably, expression levels of mitophagy-related proteins PINK1, Parkin, and LC3 increased significantly while the P62 protein expression decreased remarkably. Furthermore, Sal reversed the α-AMA-induced decrease in cell viability and mitochondrial membrane potential and increase in intracellular ROS level. In addition, Sal promoted expression levels of PINK1, Parkin, and LC3 II while suppressing the Bax/Bcl-2 ratio, Cleaved-Caspase 3, and Cleaved-PARP I as well as P62. The results above proved that salidroside alleviates α-AMA-induced mouse liver cells damage via promoting PINK1/Parkin-mediated mitophagy and reducing cell apoptosis.

Amanitin-induced variable cytotoxicity in various cell lines is mediated by the different expression levels of OATP1B3

Amanita phalloides is one of the deadliest mushrooms worldwide, causing most fatal cases of mushroom poisoning. Among the poisonous substances of Amanita phalloides, amanitins are the most lethal toxins to humans. Currently, there are no specific antidotes available for managing amanitin poisoning and treatments are lack of efficacy. Amanitin mainly causes severe injuries to specific organs, such as the liver, stomach, and kidney, whereas the lung, heart, and brain are hardly affected. However, the molecular mechanism of this phenomenon remains not understood. To explore the possible mechanism of organ specificity of amanitin-induced toxicity, eight human cell lines derived from different organs were exposed to α, β, and γ-amanitin at concentrations ranging from 0.3 to 100 μM. We found that the cytotoxicity of amanitin differs greatly in various cell lines, among which liver-derived HepG2, stomach-derived BGC-823, and kidney-derived HEK-293 cells are most sensitive. Further mechanistic study revealed that the variable cytotoxicity is mainly dependent on the different expression levels of the organic anion transporting polypeptide 1B3 (OATP1B3), which facilitates the internalization of amanitin into cells. Besides, knockdown of OATP1B3 in HepG2 cells prevented α-amanitin-induced cytotoxicity. These results indicated that OATP1B3 may be a crucial therapeutic target against amanitin-induced organ failure.

The effects of CDP-choline treatment in Amanita phalloides mushroom toxicosis

Amanita phalloides poisoning is known to be the most fatal case among mushroom poisoning cases. Its main mechanism of toxicity is that it leads to cell death by the irreversible binding of its toxins to the DNA-dependent RNA polymerase II enzyme. This study was planned to analyze the effects of the CDP-choline molecule on Amanita phalloides mushroom poisoning cases.The extract of the Amanita phalloides mushroom was taken and intraperitoneally administered to male Wistar Albino rats at a dose of 0.3 g/kg. In the experiment phase, the rats were divided into three groups of CDP-choline treatment according to the doses of 100 mg/kg, 250 mg/kg, and 500 mg/kg, and one control group was administered a 1 ml/kg dose of 0.9% isotonic NaCl solution. The treatments were then administered intraperitoneally at the 2nd hour, and at the 6th hour, the rats were sacrificed. The degree of damage in the liver and kidney tissues of the rats was evaluated histopathologically.It was concluded that CDP-choline reduced or prevented the damage that occurred in the liver significantly and dose-dependently in the toxicosis picture caused by Amanita phalloides, and it showed a tendency to lower or prevent the damage in the kidney, albeit not significantly.

Biodistribution of radiolabeled alpha-amanitin in mice: An Investigation

Mushroom poisonings caused by Amanita phalloides are the leading cause of mushroom-related deaths worldwide. Alpha-Amanitin (α-AMA), a toxic substance present in these mushrooms, is responsible for the resulting hepatotoxicity and nephrotoxicity. The objective of our study was to determine the distribution of α-AMA in Balb/c mice by labeling with Iodine-131.Mice were injected with a toxic dose (1.4 mg/kg) of α-AMA labeled with Iodine-131. The mice were sacrificed at the 1st, 2nd, 4th, 8th, 24th, and 48th hours under anesthesia. The organs of the mice were removed, and their biodistribution was assessed in all experiments. The percent injected dose per gram (ID/g %) value for kidney, liver, lung, and heart tissues at 1st hour were 1.59 ± 0.07, 1.25 ± 0.33, 3.67 ± 0.80 and 1.07 ± 0.01 respectively.This study provides insights into the potential long-term effects of α-AMA accumulation in specific organs. Additionally, this study has generated essential data that can be used to demonstrate the impact of antidotes on the biological distribution of α-AMA in future toxicity models.

Examining the associations between a generalist feeder and a highly toxic host.

Understanding the often antagonistic plant-herbivore interactions and how host defenses can influence herbivore dietary breadth is an area of ongoing study in ecology and evolutionary biology. Typically, host plants/fungi that produce highly noxious chemical defenses are only fed on by specialists. We know very little about generalist species that can feed and develop on a noxious host. One such example of generalists feeding on toxic host occurs in the mushroom-feeding Drosophila found in the immigrans-tripunctata radiation. Although these species are classified as generalists, their acceptable hosts include deadly Amanita species. In this study, we used behavioral assays to assess associations between one mushroom-feeding species, Drosophila guttifera, and the deadly Amanita phalloides. We conducted feeding assays to confirm the presence of cyclopeptide toxin tolerance. We then completed host preference assays in female flies and larvae and did not find a preference for toxic mushrooms in either. Finally, we assessed the effect of competition on oviposition preference. We found that the presence of a competitor's eggs on the preferred host was associated with the flies increasing the number of eggs laid on the toxic mushrooms. Our results highlight how access to a low competition host resource may help to maintain associations between a generalist species and a highly toxic host.

Unraveling Hematotoxicity of α-Amanitin in Cultured Hematopoietic Cells.

Amanita phalloides poisonings account for the majority of fatal mushroom poisonings. Recently, we identified hematotoxicity as a relevant aspect of Amanita poisonings. In this study, we investigated the effects of the main toxins of Amanita phalloides, α- and β-amanitin, on hematopoietic cell viability in vitro. Hematopoietic cell lines were exposed to α-amanitin or β-amanitin for up to 72 h with or without the pan-caspase inhibitor Z-VAD(OH)-FMK, antidotes N-acetylcysteine, silibinin, and benzylpenicillin, and organic anion-transporting polypeptide 1B3 (OATP1B3) inhibitors rifampicin and cyclosporin. Cell viability was established by trypan blue exclusion, annexin V staining, and a MTS assay. Caspase-3/7 activity was determined with Caspase-Glo assay, and cleaved caspase-3 was quantified by Western analysis. Cell number and colony-forming units were quantified after exposure to α-amanitin in primary CD34+ hematopoietic stem cells. In all cell lines, α-amanitin concentration-dependently decreased viability and mitochondrial activity. β-Amanitin was less toxic, but still significantly reduced viability. α-Amanitin increased caspase-3/7 activity by 2.8-fold and cleaved caspase-3 by 2.3-fold. Z-VAD(OH)-FMK significantly reduced α-amanitin-induced toxicity. In CD34+ stem cells, α-amanitin decreased the number of colonies and cells. The antidotes and OATP1B3 inhibitors did not reverse α-amanitin-induced toxicity. In conclusion, α-amanitin induces apoptosis in hematopoietic cells via a caspase-dependent mechanism.

Diagnosis and Management of Amanita Phalloides Toxicity in the Emergency Department Observation Unit: A Case Report.

Mushroom toxicity is an important etiology of acute liver injury in a patient with gastrointestinal symptoms. We present the case of a male patient presenting to the emergency department (ED) with gastrointestinal distress who was placed under ED observation for elevated liver function tests. During his hospital course, it was revealed he had consumed wild mushrooms believed to be Amanita phalloides. While mushroom ingestion and subsequent toxicity are rare, a high index of suspicion in foraging hobbyists is essential to arriving at the correct diagnosis and directing the patient to the appropriate management.

Death caps (Amanita phalloides) frequently establish from sexual spores, but individuals can grow large and live for more than a decade in invaded forests.

Global change is reshaping Earth's biodiversity, but the changing distributions of nonpathogenic fungi remain largely undocumented, as do mechanisms enabling invasions. The ectomycorrhizal Amanita phalloides is native to Europe and invasive in North America. Using population genetics and genomics, we sought to describe the life history traits of this successfully invading symbiotic fungus. To test whether death caps spread underground using hyphae, or aboveground using sexual spores, we mapped and genotyped mushrooms from European and US sites. Larger genetic individuals (genets) would suggest spread mediated by vegetative growth, while many small genets would suggest dispersal mediated by spores. To test whether genets are ephemeral or persistent, we also sampled from populations over time. At nearly every site and across all time points, mushrooms resolve into small genets. Individuals frequently establish from sexual spores. But at one Californian site, a single individual measuring nearly 10 m across dominated. At two Californian sites, the same genetic individuals were discovered in 2004, 2014, and 2015, suggesting single individuals (both large and small) can reproduce repeatedly over relatively long timescales. A flexible life history strategy combining both mycelial growth and spore dispersal appears to underpin the invasion of this deadly perennial ectomycorrhizal fungus.

Finding of the Amanita phalloides in the Akmola region and its distribution in Kazakhstan

The article presents data on a new finding of the Death cap – Amanita phalloides (Vaill. ex Fr.) Link in Kazakhstan. The species were found in two locations in a mixed forest in the vicinity of the Shchuchinsky village near the Shchuchinsk city. This is the first record of the species for the Akmola region and the fourth location in Kazakhstan. Based on the studied specimens, a clarified description of the macroscopic and microscopic characteristics of A. phalloides. Detailed descriptions for specimens from Kazakhstan are provided for the first time. A comparison of the obtained description with literary data did not reveal significant discrepancies. The photographs of the macro- and microcharacteristics of the collected specimens are presented in the article. A number of literary and electronic sources were analyzed, as a result of which a map of the Death cap finds in Kazakhstan and in the adjacent territory of Russia was built. Considering its general distribution, the discovery of the A. phalloides can be expected in other forested areas of the northern and eastern regions of the country in the future. The article discusses the overall distribution of this species in the republic and the likelihood of its appearance as a result of introduction. 
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Invasive Californian death caps develop mushrooms unisexually and bisexually

Canonical sexual reproduction among basidiomycete fungi involves the fusion of two haploid individuals of different mating types, resulting in a heterokaryotic mycelial body made up of genetically different nuclei. Using population genomics data and experiments, we discover mushrooms of the invasive and deadly Amanita phalloides can also be homokaryotic; evidence of sexual reproduction by single, unmated individuals. In California, genotypes of homokaryotic mushrooms are also found in heterokaryotic mushrooms, implying nuclei of homokaryotic mycelia are also involved in outcrossing. We find death cap mating is controlled by a single mating type locus, but the development of homokaryotic mushrooms appears to bypass mating type gene control. Ultimately, sporulation is enabled by nuclei able to reproduce alone as well as with others, and nuclei competent for both unisexuality and bisexuality have persisted in invaded habitats for at least 17 but potentially as long as 30 years. The diverse reproductive strategies of invasive death caps are likely facilitating its rapid spread, suggesting a profound similarity between plant, animal and fungal invasions.

Distribution changes of the toxic mushroom Amanita phalloides under climate change scenarios and its potential risk over indigenous communities in Mexico

Amanita phalloides is a native European deathly ectomycorrhizal mushroom that was introduced to North America and has been expanding its distribution during the last decades. This species is morphologically similar to wild edible mushrooms and if its distribution expands to Mexico, it could represent a risk in terms of food security for local communities. The aim of this study was to evaluate the potential climatic suitability that exists for A. phalloides in North America and overlay it with the distribution of mycophilic communities in Mexico under a baseline climatic scenario and climate change scenarios. To find climatic suitability we modeled its potential distribution with the algorithm that had the best predictive power after pilot test (MaxEnt) using species presences and eight climatic variables chosen with biological and statistical criteria. We worked with CanESM5 because it is one of the best models to simulate climate in North America and SSP5-8.5 scenario in order to be consistent with the precautionary principle. Our results suggest that even when the species has not yet been registered in Mexico, when using European records to model, this country presents 33.61% of climatic suitability for this species under the baseline scenario, potentially affecting about 70% of indigenous communities which are the main consumers of edible mushrooms. Under climate change scenarios, an increase in climatic suitability is expected in Mexico, while decreases are expected in United States and Canada. When using North American records to model, almost no climatic suitability is found in Mexico; however, the implementation of warning campaigns in Mexico is still needed.

Implementation of Deep Learning for Classification of Mushroom Using CNN Algorithm

Mushrooms are a type of low-level plant that lacks chlorophyll. One of the advantages of fungi is that they are commonly utilized as food items in the community. This paper discussed the implementation of CNN for the classification of mushrooms. The project aims to develop a robust system that can automate the labor-intensive task of mushroom classification. The CNN model will be trained on a large dataset of annotated mushroom images, learning to extract meaningful features and patterns for accurate categorization. To evaluate the performance of the developed system, a comprehensive set of metrics, including accuracy, precision, recall, and F1 score, will be used. The dataset will be split into training, validation, and testing sets to assess the model's generalization ability to unseen data. Based on the experimental result, the average accuracy rate in the Agaricus Portobello test was % -99.89 %, % -99.89 % in the Amanita Phalloides test, % -99.59 % in the Cantharellus Cibarius test, % -98.89 % in the Gyromitra Esculenta test, % -99.96 % in the Hygrocybe Conica, and % -99.93 % in the Omphalotus Orealius.

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<i>Amanita phalloides</i> in Northwest European Russia

The article reviews available data on the “death cap” (Amanita phalloides) occurrences in the North-West of the European Russia (Leningrad, Novgorod, Pskov Oblasts and St. Petersburg City). The literature data are analyzed, the review of ecological preferences of the species in the studied area is carried out. It has been suggested that the species can be both native and imported and was introduced with broad-leaved tree seedlings during the foundation of numerous manor parks in the XVIII–XIX centuries. Molecular-genetic analysis was carried out, showing that ITS sequences of A. phalloides samples from Leningrad Oblast form a common clade with the sequences of collections from Central Russia as well as from Northern and Central Europe.

α-amanitin induces autophagy through AMPK-mTOR-ULK1 signaling pathway in hepatocytes

Amanitin poisoning is one of the most life-threatening mushroom poisonings. α-Amanitin plays a key role in Amanita phalloides intoxication. α-Amanitin shows toxic effects on the liver. However, the mechanism by which α-amanitin induces liver injury has not been elucidated. Autophagy plays a crucial role in maintaining cellular homeostasis and is closely related to the occurrence of a variety of diseases. Studies have shown that autophagy may play an important role in the process of α-amanitin-induced liver injury. However, the mechanism of α-amanitin-induced autophagy remains unclear. Thus, this study aimed to explore the mechanisms of α-amanitin in inducing hepatotoxicity in Sprague Dawley (SD) rats and the normal human liver cell line L02 cells. The SD rats and L02 cells exposed to α-amanitin were observed to determine whether α-amanitin could induce the autophagy of rat liver and L02 cells. The regulatory relationship between autophagy and the AMPK-mTOR-ULK pathway by exposing the autophagy agonist (rapamycin (RAPA)), autophagy inhibitor (3-methylademine (3-MA)), and AMPK inhibitor (compound C) was also explored. Autophagy-related proteins and AMPK-mTOR-ULK pathway-related proteins were detected using Western blot. The results of the study indicated that exposure to different concentrations of α-amanitin led to morphological changes in liver cells and significantly elevated levels of ALT and AST in the serum of SD rats. Additionally, the expression levels of LC3-II, Beclin-1, ATG5, ATG7, AMPK, p-AMPK, mTOR, p-mTOR, and ULK1 were significantly increased in the rat liver. And we found that L02 cells exposed to 0.5 μM α-amanitin for 6 h significantly induced autophagy and activated the AMPK-mTOR-ULK1 pathway. Pretreated with RAPA, 3-MA, and compound C for 1 h, the expression levels of autophagy-related proteins and AMPK-mTOR-ULK pathway-related proteins significantly changed. Our results indicates that autophagy and the AMPK-mTOR-ULK pathway are involved in the process of α-amanitin-induced liver injury. This study may foster the identification of actionable therapeutic targets for A. phalloides intoxication.

Pangenomics of the death cap mushroom Amanita phalloides, and of Agaricales, reveals dynamic evolution of toxin genes in an invasive range

The poisonous European mushroom Amanita phalloides (the “death cap”) is invading California. Whether the death caps’ toxic secondary metabolites are evolving as it invades is unknown. We developed a bioinformatic pipeline to identify the MSDIN genes underpinning toxicity and probed 88 death cap genomes from an invasive Californian population and from the European range, discovering a previously unsuspected diversity of MSDINs made up of both core and accessory elements. Each death cap individual possesses a unique suite of MSDINs, and toxin genes are significantly differentiated between Californian and European samples. MSDIN genes are maintained by strong natural selection, and chemical profiling confirms MSDIN genes are expressed and result in distinct phenotypes; our chemical profiling also identified a new MSDIN peptide. Toxin genes are physically clustered within genomes. We contextualize our discoveries by probing for MSDINs in genomes from across the order Agaricales, revealing MSDIN diversity originated in independent gene family expansions among genera. We also report the discovery of an MSDIN in an Amanita outside the “lethal Amanitas” clade. Finally, the identification of an MSDIN gene and its associated processing gene (POPB) in Clavaria fumosa suggest the origin of MSDINs is older than previously suspected. The dynamic evolution of MSDINs underscores their potential to mediate ecological interactions, implicating MSDINs in the ongoing invasion. Our data change the understanding of the evolutionary history of poisonous mushrooms, emphasizing striking parallels to convergently evolved animal toxins. Our pipeline provides a roadmap for exploring secondary metabolites in other basidiomycetes and will enable drug prospecting.