Fatal Mushroom Poisonings Research Articles

Investigation of a family cluster poisoning incident caused by Galerina sulciceps mushroom

Mushroom poisoning is the most important cause of death in food-borne poisoning in China, mainly caused by amanitin, which is caused by rapid progression, complex mechanism and latency. Early identification, diagnosis and treatment are important to improve the prognosis of fatal mushroom poisoning. This article analyzes the clinical characteristics, identification process and treatment of 14 patients with amanitin-containing Galerina sulciceps mushroom poisoning in a family, so as to improve the identification ability of the first physician in recognizing and managing early-stage mushroom poisoning, and to increase the cure rate through early bundle therapy of mushroom poisoning.

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.

Characterization of Lethal Toxin-Producing Genes in Amanita brunneitoxicaria and PCR-based Detection of Deadly Poisonous Amanitas in the Section Phalloideae

A recent discovery of Amanita brunneitoxicaria revealed that this mushroom and A. exitialis are the main causative agents of fatal mushroom poisoning in Thailand. Poisonous amatoxins present are bicyclic octapeptides encoded by the MSDIN family and macrocyclized by prolyl oligopeptidase B enzyme. Diversity of the MSDIN toxin-associated genes in A. brunneitoxicaria however, remains unexplored. Therefore, this study aimed to characterize the MSDIN family members in A. brunneitoxicaria in comparison with A. exitialis based on whole genome sequencing, followed by gene annotation using bioinformatic tools. Identification of conserved core peptides of the MSDIN family revealed the presence of α-amanitin and β-amanitin as endogenous toxins in A. brunneitoxicaria. In addition to the major amatoxins, amanexitide and phallacidin were also found in A. exitialis. Phylogenetic analyses of the MSDIN family members showed that an unknown peptide present in A. brunneitoxicaria is clustered with phalloidin. On the basis of PCR detection, a modified forward primer was incorporated into the PCR reactions to obtain clear and distinctive DNA fragments of approximately 300 bp from three deadly poisonous mushrooms of A. brunneitoxicaria, A. exitialis and A. fuliginea in the section Phalloideae. Hence, they could be distinguished from other edible and less toxic species in the same genus. Such rapid and accurate identification of the target region of the MSDIN family from the lethal mushrooms is relevant as means to reduce mortality. HIGHLIGHTS Amanita brunneitoxicaria and A. exitialis are responsible for most cases of fatal mushroom poisoning in Thailand During wild mushroom foraging, A. brunneitoxicaria is often mistaken for the edible species of A. vaginata A. brunneitoxicaria and other lethal aminitas contain amatoxin-producing genes encoded by the MSDIN family PCR-based detection of the target region of the MSDIN family can be used to distinguish the lethal aminitas in the section Phalloideae This rapid detection is useful for timely and appropriate medical treatment of mushroom intoxication GRAPHICAL ABSTRACT

Occurrence and chemotaxonomical analysis of amatoxins in Lepiota spp. (Agaricales)

About 95% of fatal mushroom poisonings worldwide are caused by amatoxins and phallotoxins mostly produced by species of Amanita, Galerina, and Lepiota. The genus Lepiota is supposed to include a high number of species producing amatoxins. In this study, we investigated 16 species of Lepiota based on 48 recently collected specimens for the presence of amatoxins by liquid chromatography coupled to a diode-array detector and mass spectrometry (UHPLC-QTOF-MS/MS). By comparing the retention times, UV absorptions, and diagnostic MS fragment ions with data obtained from the benchmark species Amanita phalloides, we detected α-amanitin and γ-amanitin in Lepiota subincarnata, α-amanitin and amaninamide in Lepiota brunneoincarnata, and β-amanitin and α-amanitin in Lepiota elaiophylla. Phallotoxins have not been detected any of these species. Two possibly undescribed amatoxin derivatives were found in Lepiota boudieri and L. elaiophylla, as well as one further non-amatoxin compound in one specimen of L. cf. boudieri. These compounds might be used to differentiate L. elaiophylla from L. xanthophylla and species within the L. boudieri species complex. No amatoxins were detected in L. aspera, L. castanea, L. clypeolaria, L. cristata, L. erminea, L. felina, L. fuscovinacea, L. lilacea, L. magnispora, L. oreadiformis, L. pseudolilacea, L. sp. (SeSa 5), and L. subalba. By combining the occurrence data of amatoxins with a phylogenetic analysis, a monophyletic group of amatoxin containing species of Lepiota is evident. These chemotaxonomic results highlight the relevance of systematic relationships for the occurrence of amatoxins and expand our knowledge about the toxicity of species of Lepiota.

Cyclopeptide toxins of lethal amanitas: Compositions, distribution and phylogenetic implication

Lethal amanitas (Amanita sect. Phalloideae) are responsible for 90% of all fatal mushroom poisonings. Since 2000, more than ten new lethal Amanita species have been discovered and some of them had caused severe mushroom poisonings in China. However, the contents and distribution of cyclopeptides in these lethal mushrooms remain poorly known. In this study, the diversity of major cyclopeptide toxins in seven Amanita species from Eastern Asia and three species from Europe and North America were systematically analyzed, and a new approach to inferring phylogenetic relationships using cyclopeptide profile was evaluated for the first time. The results showed that there were diversities of the cyclopeptides among lethal Amanita species, and cyclopeptides from Amanita rimosa and Amanita fuligineoides were reported for the first time. The amounts of amatoxins in East Asian Amanita species were significantly higher than those in European and North American species. The analysis of distribution of amatoxins and phallotoxins in various Amanita species demonstrated that the content of phallotoxins was higher than that of amatoxins in Amanita phalloides and Amanita virosa. In contrast, the content of phallotoxins was significantly lower than that of amatoxins in all East Asian lethal Amanita species tested. However, the distribution of amatoxins and phallotoxins in different tissues showed the same tendency. Eight cyclopeptides and three unknown compounds were identified using cyclopeptide standards and high-resolution MS. Based on the cyclopeptide profiles, phylogenetic relationships of lethal amanitas were inferred through a dendrogram generated by UPGMA method. The results showed high similarity to the phylogeny established previously based on the multi-locus DNA sequences.

Hepatoprotective Effects and Mechanisms of Action of Triterpenoids from Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) on α-Amanitin-Induced Liver Injury in Mice.

Most fatal mushroom poisonings are caused by species of the genus Amanita; the amatoxins are responsible for acute liver failure and death in humans. Ganoderma lucidum is a well-known traditional medicinal mushroom that has been shown to have obvious hepatoprotective effects. This study evaluated the hepatoprotective effects of triterpenoids from G. lucidum on liver injury induced by a-amanitin (α-AMA) in mice and the mechanisms of action of these triterpenoids, including radical scavenging and antiapoptosis activities. Mice were treated with α-AMA, followed by G. lucidum total triterpenoids or individual triterpenoids, and their hepatoprotective effects were compared with those of the reference drug silibinin (SIL). Treatment with SIL, G. lucidum total triterpenoids, and each of the 5 individual triterpenoids significantly reduced serum alanine aminotransaminase and aspartate ami- notransaminase concentrations and reduced mortality rates 20-40%. Moreover, triterpenoids and SIL significantly enhanced superoxide dismutase and catalase activity and reduced malondialdehyde content in livers. Treatment with ganoderic acid C2 significantly inhibited DNA fragmentation and decreased caspase-3, -8, and -9 activities. The results demonstrated that triterpenoids have hepatoprotective effects on α-AMA-induced liver injury and that their hepatoprotective mechanisms may be the result of their antioxidative and radical scavenging activities and their inhibition of apoptosis.

Production of (15)N-labeled α-amanitin in Galerina marginata.

α-Amanitin is the major causal constituent of deadly Amanita mushrooms that account for the majority of fatal mushroom poisonings worldwide. It is also an important biochemical tool for the study of its target, RNA polymerase II. The commercial supply of this bicyclic peptide comes from Amanita phalloides, the death cap mushroom, which is collected from the wild. Isotopically labeled amanitin could be useful for clinical and forensic applications, but α-amanitin has not been chemically synthesized and A. phalloides cannot be cultured on artificial medium. Using Galerina marginata, an unrelated saprotrophic mushroom that grows and produces α-amanitin in culture, we describe a method for producing (15)N-labeled α-amanitin using growth media containing (15)N as sole nitrogen source. A key to success was preparing (15)N-enriched yeast extract via a novel method designated "glass bead-assisted maturation." In the presence of the labeled yeast extract and (15)N-NH4Cl, α-amanitin was produced with >97% isotope enrichment. The labeled product was confirmed by HPLC, high-resolution mass spectrometry, and NMR.

DETERMINATION OF MUSHROOM TOXIN ALPHA-AMANITIN IN SERUM BY LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY AFTER SOLID-PHASE EXTRACTION

Alpha-amanitin is a cyclic peptide which belongs to a large group of mushroom toxins known as amatoxins. Being responsible for the majority of fatal mushroom poisonings, they require rapid detection and excretion from the body fluids. In accordance with these requirements, a simple and an accurate method was developed for successful identification and quantification of alpha-amanitin in serum with electrospray liquid chromatography–mass spectrometry (LC-ESI-MS) after collision-induced dissociation. The method conforms to the established International Conference on Harmonization Q2A/Q2B 1996 guidelines on the validation of analytical methods. Linearity, precision, extraction recovery and stability test on blank serum spiked with alpha-amanitin and stored in different conditions met the acceptance criteria. The obtained calibration curve was linear over the concentration range 5-100 ng/mL with a lower limit of quantification (LOQ) of 5 ng/mL and limit of detection (LOD) of 2.5 ng/mL. The mean intraand inter-day precision and accuracy were 6.05% and less than ±15% of nominal values, respectively. The neutral solid phase extraction with copolymer hydrophilic–lipophilic balance cartridges was found optimal for sample preparation with the mean recovery of 91.94%. The proposed method demonstrated high sensitivity and selectivity which can be useful both for clinical and forensic toxicology analysis of alpha-amanitin at low concentrations. Acta Medica Medianae 2015;54(1): 12-19.

Peptide Macrocyclization Catalyzed by a Prolyl Oligopeptidase Involved in α-Amanitin Biosynthesis

Amatoxins are ribosomally encoded and posttranslationally modified peptides that account for the majority of fatal mushroom poisonings of humans. Arepresentative amatoxin is the bicyclic octapeptideα-amanitin, formed via head-to-tail macrocyclization, which is ribosomally biosynthesized as a 35-amino acid propeptide in Amanita bisporigera and in the distantly related mushroom Galerina marginata. Although members of the prolyl oligopeptidase (POP) family of serine proteases have been proposed to play a role in α-amanitin posttranslational processing, the exact mechanistic details are not known. Here, we show that a specific POP (GmPOPB) is required for toxin maturation in G.marginata. Recombinant GmPOPB catalyzed two nonprocessive reactions: hydrolysis at an internal Pro to release the C-terminal 25-mer from the 35-mer propeptide and transpeptidation at the second Pro to produce the cyclic octamer. On the other hand, we show thatGmPOPA, the putative housekeeping POP of G.marginata, behaves like a conventional POP.

Profiling of amatoxins and phallotoxins in the genus Lepiota by liquid chromatography combined with UV absorbance and mass spectrometry.

Species in the mushroom genus Lepiota can cause fatal mushroom poisonings due to their content of amatoxins such as α-amanitin. Previous studies of the toxin composition of poisonous Lepiota species relied on analytical methods of low sensitivity or resolution. Using liquid chromatography coupled to UV absorbance and mass spectrometry, we analyzed the spectrum of peptide toxins present in six Italian species of Lepiota, including multiple samples of three of them collected in different locations. Field taxonomic identifications were confirmed by sequencing of the internal transcribed spacer (ITS) regions. For comparison, we also analyzed specimens of Amanita phalloides from Italy and California, a specimen of A. virosa from Italy, and a laboratory-grown sample of Galerina marginata. α-Amanitin, β-amanitin, amanin, and amaninamide were detected in all samples of L. brunneoincarnata, and α-amanitin and γ-amanitin were detected in all samples of L. josserandii. Phallotoxins were not detected in either species. No amatoxins or phallotoxins were detected in L. clypeolaria, L. cristata, L. echinacea, or L. magnispora. The Italian and California isolates of A. phalloides had similar profiles of amatoxins and phallotoxins, although the California isolate contained more β-amanitin relative to α-amanitin. Amaninamide was detected only in A. virosa.

Clinical importance of toxin concentration in Amanita verna mushroom

Poisoning from Amanita group of mushrooms comprises approximately 3% of all poisonings in our country and their being responsible for nearly the entire fatal mushroom poisonings makes them important. These mushrooms contain primarily two types of toxins, amatoxins and phallotoxins. Phallotoxins have a more limited toxicity potential and they primarily consist of phalloidin (PHN) and phallacidin (PCN). Amatoxins, on the other hand, are very toxic and they primarily consist of alpha-amanitin (AA), beta-amanitin (BA) and gamma-amanitin (GA). Toxin levels can vary among various species, even among varieties of the same species, of Amanita mushroom family. Revealing the differences between the toxin compositions of the Amanita species that grow in our region may contribute to the clinics of poisonings. Our study aims at showing in detail the toxin levels in various parts of Amanita verna mushroom.A. verna mushrooms needed for toxin analysis were collected from Kozak Plateau near Ayvalik county of Balıkesir, Turkey in April 2013. The mushrooms were divided into their parts as pileus, gills, stripe and volva. Following the procedures required before the analysis, the AA, BA, GA, PHN and PCN levels were measured using the RP-HPLC method.While the lowest level of amatoxin was in the volva of the mushroom, the highest was measured in the gills. This was followed by pileus and stripe where the levels were close to each other. Similarly, the highest level of phallotoxin was measured in the gills. Gamma toxin and phalloidin were at lower amounts than the other toxins.A. verna is frequently confused with edible mushrooms with white caps due to its macroscopic similarity. If just one of them is eaten by mistake by an adult person with no mushroom experience, it can easily poison them. The amount of amatoxin is more as compared to Amanita phalloides and A. phalloides var. alba. Particularly, the AA and BA levels are approximately three times higher, whereas GA levels are lower. Similarly, the level of PCN is approximately four times higher as compared to A. phalloides and A. phalloides var. alba; by contrast, the level of PNH is about a half of theirs.In summary, it can be said that A. verna is a more toxic mushroom than A. phalloides and has a higher rate of mortality. With our study, the amatoxin and phallotoxin concentrations and distribution in A. verna mushrooms were shown in detail for the first time and it would be useful to carry out more similar studies with other members of Amanita family growing in various parts of the world.

Acute liver failure after amanitin poisoning: a porcine model to detect prognostic markers for liver regeneration.

Over 90% of fatal mushroom poisoning occurs after ingestion of amanitin-containing species. This study aimed to investigate markers indicating spontaneous liver regeneration in a porcine acute liver failure (ALF) model after α-amanitin intoxication. German landrace pigs received either 0.15mg/kg (n=5) α-amanitin intravenously or 0.35mg/kg (n=5) intraportally. Pigs were invasively monitored and kept under general anesthesia throughout the experiment. Laboratory parameters were analyzed every 8h. ALF occurred in all animals (10/10) 41±3h after intoxication. All pigs receiving 0.35mg/kg α-amanitin and one pig receiving 0.15mg/kg α-amanitin died 57±16h after the primary onset of ALF. Four pigs of the 0.15mg/kg intoxication group recovered spontaneously from ALF after 56±6h. Starting at 32h after intoxication, significantly higher values of albumin and total plasma protein could be measured in surviving animals (p<0.05). A significant temporary increase in the tumor necrosis factor alpha (TNF-α) plasma concentration was detected 40-80h after intoxication in recovering animals (p<0.05). This porcine model represents a novel tool to analyse multiple aspects of liver regeneration following α-amanitin poisoning to allow early discrimination between a fatal course and survivors. Decreased albumin and total plasma protein concentrations in the early intoxication phase indicated a lethal outcome, while an increase in the TNF-α plasma concentration was identified as the earliest prognostic plasma marker detecting liver regeneration a long time before liver function was biochemically and clinically impaired.

Legalon® SIL: The Antidote of Choice in Patients with Acute Hepatotoxicity from Amatoxin Poisoning

More than 90% of all fatal mushroom poisonings worldwide are due to amatoxin containing species that grow abundantly in Europe, South Asia, and the Indian subcontinent. Many cases have also been reported in North America. Initial symptoms of abdominal cramps, vomiting, and a severe cholera-like diarrhea generally do not manifest until at least six to eight hours following ingestion and can be followed by renal and hepatic failure. Outcomes range from complete recovery to fulminant organ failure and death which can sometimes be averted by liver transplant. There are no controlled clinical studies available due to ethical reasons, but uncontrolled trials and case reports describe successful treatment with intravenous silibinin (Legalon® SIL). In nearly 1,500 documented cases, the overall mortality in patients treated with Legalon® SIL is less than 10% in comparison to more than 20% when using penicillin or a combination of silibinin and penicillin. Silibinin, a proven antioxidative and anti-inflammatory acting flavonolignan isolated from milk thistle extracts, has been shown to interact with specific hepatic transport proteins blocking cellular amatoxin re-uptake and thus interrupting enterohepatic circulation of the toxin. The addition of intravenous silibinin to aggressive intravenous fluid management serves to arrest and allow reversal of the manifestation of fulminant hepatic failure, even in severely poisoned patients. These findings together with the available clinical experience justify the use of silibinin as Legalon® SIL in Amanita poisoning cases.

Early morphological and functional alterations in canine hepatocytes due to α-amanitin, a major toxin of Amanita phalloides

The toadstool death cap (Amanita phalloides) and its subspecies, destroying angel (A. virosa) and death angel (A. verna) are responsible for nearly 95% of all fatal mushroom poisonings. High mortality rate in A. phalloides intoxications is principally a result of the acute liver failure following significant hepatocyte damage due to hepatocellular uptake of amanitins, the major toxins of this mushroom. This study evaluated early morphological and functional alterations in hepatocytes exposed to different concentrations of alpha-amanitin (alpha-AMA). All experiments were performed on cultured canine hepatocytes since intoxicated with A. phalloides dogs have clinical course and pathological findings similar to those seen in humans. The overall functional integrity and viability of cultured hepatocytes were assessed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and by measurements of lactate dehydrogenase (LDH), total protein, and urea levels. Our results showed that the course of alpha-AMA toxicity in cultured dog hepatocytes is divided into two phases. The first phase comprises functional cell impairments expressed by significant increase of LDH activity and inhibition of protein and urea synthesis when compared with the control group. This is followed by discrete changes in hepatocyte ultrastructure, including marginalization and condensation of nuclear chromatin, as well as formation of the foamlike cytoplasm. The second stage is lethal and is characterized by ongoing necrosis, and/or apoptosis. This may be related to dose of toxin and time of exposure.

Syndromic diagnosis and management of confirmed mushroom poisonings

To assess the evolving global epidemiology of mushroom poisoning and to identify new and emerging mushroom poisonings and their treatments, a descriptive analysis and review of the world's salient scientific literature on mushroom poisoning was conducted. Data sources from observation studies conducted over the period 1959-2002, and describing 28,018 mushroom poisonings since 1951, were collected from case reports, case series, regional descriptive studies, meta-analyses, and laboratory studies of mushroom poisonings and the toxicokinetics of mycotoxins. Studies included in the review were selected by a MEDLINE search, 1966-2004, an Ovid OLDMEDLINE search, 1951-1965, and a medical library search for sources published before 1951. To better guide clinicians in establishing diagnoses and implementing therapies, despite confusing ingestion histories, data were extracted to permit an expanded syndromic classification of mushroom poisoning based on presentation timing and target organ systemic toxicity. The final 14 major syndromes of mushroom poisoning were stratified first by presentation timing and then by target organ systemic toxicity and included early (<6 hrs), late (6-24 hrs), and delayed syndromes (> or =1 day). There were eight early syndromes (four neurotoxic, two gastrointestinal, two allergic); three late syndromes (hepatotoxic, accelerated nephrotoxic, erythromelalgia); and three delayed syndromes (delayed nephrotoxic, delayed neurotoxic, rhabdomyolysis). Four new mushroom poisoning syndromes were classified including accelerated nephrotoxicity (Amanita proxima, Amanita smithiana), rhabdomyolysis (Tricholoma equestre, Russula subnigricans), erythromelalgia (Clitocybe amoenolens, Clitocybe acromelalgia), and delayed neurotoxicity (Hapalopilus rutilans). In addition, data sources were stratified by three chronological time periods with >1,000 confirmed mushroom ingestions reported and tested for any statistically significant secular trends in case fatalities from mushroom ingestions over the entire study period, 1951-2002. Since the 1950s, reports of severe and fatal mushroom poisonings have increased worldwide. Clinicians must consider mushroom poisoning in the evaluation of all patients who may be intoxicated by natural substances. Since information on natural exposures is often insufficient and incorrect, a new syndromic classification of mushroom poisoning is recommended to guide clinicians in making earlier diagnoses, especially in cases where only advanced critical care, including organ transplantation, may be life saving.

Evolving global epidemiology, syndromic classification, general management, and prevention of unknown mushroom poisonings

To assess the evolving global epidemiology of mushroom poisoning and to identify new and emerging mushroom poisonings and their treatments, a descriptive analysis and review of the world's salient scientific literature on mushroom poisoning was conducted. Data sources from observation studies conducted over the period 1959-2002 and describing 28,018 mushroom poisonings since 1951 were collected from case reports, case series, regional descriptive studies, meta-analyses, and laboratory studies of mushroom poisonings and the toxicokinetics of mycotoxins. Studies included in the review were selected by a MEDLINE search, 1966-2004, an Ovid OLDMEDLINE search, 1951-1965, and a medical library search for sources published before 1951. To better guide clinicians in establishing diagnoses and implementing therapies, despite confusing ingestion histories, data were extracted to permit an expanded syndromic classification of mushroom poisoning based on presentation timing and target organ systemic toxicity. The final 14 major syndromes of mushroom poisoning were stratified first by presentation timing and then by target organ systemic toxicity and included early (<6 hrs), late (6-24 hrs), and delayed syndromes (> or =1 day). There were eight early syndromes (four neurotoxic, two gastrointestinal, two allergic); three late syndromes (hepatotoxic, accelerated nephrotoxic, erythromelalgia); and three delayed syndromes (delayed nephrotoxic, delayed neurotoxic, rhabdomyolysis). Four new mushroom poisoning syndromes were classified including accelerated nephrotoxicity (Amanita proxima, Amanita smithiana), rhabdomyolysis (Tricholoma equestre, Russula subnigricans), erythromelalgia (Clitocybe amoenolens, Clitocybe acromelalgia), and delayed neurotoxicity (Hapalopilus rutilans). In addition, data sources were stratified by three chronological time periods with >1,000 confirmed mushroom ingestions reported and tested for any statistically significant secular trends in case fatalities from mushroom ingestions over the entire study period, 1951-2002. Since the 1950s, reports of severe and fatal mushroom poisonings have increased worldwide. Clinicians must consider mushroom poisoning in the evaluation of all patients who may be intoxicated by natural substances. Since information on natural exposures is often insufficient and incorrect, a new syndromic classification of mushroom poisoning is recommended to guide clinicians in making earlier diagnoses, especially in cases where only advanced critical care, including organ transplantation, may be life saving.

Liver Necrosis Caused by Mushroom Poisoning in Dogs

Mushrooms in Category A cause hepatic and renal damage after a prolonged latent period and are the most common cause of fatal mushroom poisonings in human beings. Category A is subdivided into 2 groups on the basis of the type of toxin produced. The genera Amanita and Galerina produce cyclopeptide toxins (Group I), and Gyromitra produces monomethylhydrazine toxins (Group II). Several Amanita species contain 3 kinds of cyclopeptide toxins: phallolysin, phallotoxins, and amanitins. 9 Of these toxins, only the amanitins play a significant role in poisonings. Mushrooms in categories B, C, and D have a rapid onset of clinical signs and effect the autonomic nervous system, central nervous system, and gastrointestinal tracts, respectively. Only one of the reported cases of mushroom poisoning in the dog was associated with a mushroom in Category A in which several animals were poisoned by ingesting Amanita phalloides. 6 Other mushrooms in this genus and category that have either been associated with human poisonings or contain amanitins are Amanita verna, Amanita virosa, Amanita bisporigera, and Amanita ocreata. 8 The purpose of this report is to describe the clinical and laboratory findings in 2 cases of canine mushroom poisoning. A 7-week-old male golden retriever had a sudden onset of extreme listlessness, ataxia, and petit mal seizures. The owner had observed the animal eating mushrooms on the day prior to onset of clinical signs. The puppy did not respond to intravenous Lactated Ringer’s solution and Valium and died within 6 hr of onset of clinical signs. Fresh and formalinfixed tissue, stomach contents, urine, clotted blood, and a mushroom were submitted to the diagnostic laboratory for evaluation. Strychnine was not detected in urine or stomach contents. The mushroom was identified as A. verna by a mycologist. The owner of the second case owned a small kennel and observed a sudden onset of yelping and a tense abdomen in a 12-week-old female Labrador retriever. The puppy became

Amanita Phalloides in the Eastern United States

SUMMARY Amanita phalloides has recently been collected in four eastern states: Delaware, New Jersey, New York and Virginia. Color and habit of the carpophores matched illustrations of European A. phalloides. Dimensions of spores from both the American and European specimens varied from one carpophore to another but fell in the same range. The average size and shape of spores deposited from individual specimens of A. phalloides changed as the carpophore aged, the length/width ratio passing through a maximum. Amanita phalloides was probably involved in two cases of fatal mushroom poisoning in New Jersey in November, 1970.

Mushroom Poisoning-an overview

There are many thousands of mushroom species in the world, some are edible and some are poisonous due to containing significant toxins. The edible mushroom is a common food item with tempting flavour, taste and nutritive value; nowadays quite often grown at home and cultured with commercial marketing. Mushroom poisoning usually results from ingestion of wild mushrooms due to misidentification of a toxic mushroom as an edible species bearing very close resemblance, deliberate seeking of psychotropic mushrooms and accidental childhood ingestions. Majority of fatal mushroom poisoning occurs due to ingestion of Amanita Phalloides - the 'death cap', due to its high content of Amatoxin - a potent cytotoxin. Fatal poisoning is usually associated with delayed onset of symptoms which are very severe, with hepatic, renal, hemolytic and CNS involvement. Aim of this article is for informational and preventive purpose. Key words: Mushroom poisoning; amanita phalloidis; amatoxin. DOI: 10.3126/jcmsn.v6i2.3619 Journal of college of Medical Sciences-Nepal, 2010, Vol.6, No-2, 56-61

The toxic peptides of Amanita phalloides.

Fatal mushroom poisoning is almost exclusively attributable to members of the genus Amanita. In the United States, it is Amanita verna, the so called “destroying angel”, which along with another poisonous species, A. tenuifolia, appears to play the major part in lethal mushroom poisoning (1, 2). Quite recently we were able to demonstrate the presence of the two main amanita toxins in a European sample of A. verna (73). Besides these mushrooms some relatively rare Galerina species also contain the amanita toxins (42). In Central Europe the predominant culprit is the greenish Amanita phalloides (Fig. 1). This mushroom, also known as the “deadly agaric”, is frequently confused with the delicious field mushroom Agaricus campestris or with the yellow Amanita mappa (citrina), which contains no toxic peptides, but the relatively untoxic bufotenine (5-hydroxy-N-dimethyl-tryptamine) (67, 43), a base occurring in toads and recently isolated also from different plants, among them Piptadenia peregrina (12), P. macrocarpa and P. excelsa (20), Desmodium pulchellum (19) or from Epena, a drug from a woody South American Leguminosa (28).KeywordsAspartic AcidUltraviolet SpectrumCysteic AcidPaper ElectrophoresisPaper ChromatogramThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.