Using MALDI-ToF mass spectrometry to identify mushroom species: Proof of concept analysis of Amanita genus specimens.

Food poisoning caused by toxic mushrooms, such as species in the Amanita genus, occurs frequently around the world. To properly treat these patients, it is important to rapidly and accurately identify the causal species. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectrometry is a rapid technique that has been used in medical laboratories for the past three decades to identify bacteria, yeasts, and filamentous fungi.Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-Tof MS) is a rapid method used for the past three decades to identify microorganisms. In this study, we created and internally validated a MALDI-Tof MS reference database comprising 15 Amanita species frequently encountered in France, and we challenged this database with 38 Amanita specimens from four French locations, using a free online application for MALDI-ToF spectra identifications.Assessment of the database showed that mass spectra can be obtained by analyzing any portion of a carpophore and that all portions enabled identification of the carpophore at the species level. Most carpophores were correctly identified using our database, with the exception of specimens from the Vaginatae section. Decay tests also demonstrated that decayed portions (like those found in the kitchen garbage can) of Amanita phalloides mushrooms could be properly identified using MALDI-ToF MS.Our findings provide important insight for toxicology laboratories that often rely on DNA sequencing to identify meal leftovers implicated in food poisoning. In future developments, this technique could also be used to detect counterfeit mushrooms by including other genera in the reference database. LAY SUMMARY: MALDI-ToF MS is a powerful identification tool for microorganisms. We demonstrate that the technique can be applied to Amanita specimens. This will prevent food intoxications as a rapid and definite identification can be obtained, and it can also be used for food remnants.

Fatal Amanita muscaria poisoning in a dog confirmed by PCR identification of mushrooms.

Diagnosing mushroom poisoning in dogs can be difficult and often includes identification of suspect mushrooms. Visual identification may be hindered by mastication, oral medications, or poor quality of environmental mushroom samples. Other analytical techniques may thus be necessary to aid in mushroom identification. A 5-y-old neutered male Labrador Retriever dog developed acute onset of vomiting, diarrhea, tremors, seizures, and somnolence. The dog was treated at a veterinary clinic and was briefly stabilized, but died during transport to an emergency clinic. On postmortem examination at the University of Kentucky Veterinary Diagnostic Laboratory, the dog's stomach was full of mushrooms covered with activated charcoal. Mushrooms were damaged, fragmented, and discolored, precluding accurate visual identification. Mushroom pieces were sent to the Department of Plant Pathology at the University of California-Davis for PCR identification; the neurotoxic mushroom Amanita muscaria was identified. A qualitative liquid chromatography-mass spectrometry (LC-MS) method was developed to detect ibotenic acid and muscimol, the toxic compounds present in A. muscaria. Mushrooms, stomach contents, and urine were analyzed by LC-MS; ibotenic acid and muscimol were detected in all samples. Because identification of ingested mushrooms is sometimes necessary to confirm mushroom poisoning, PCR can identify ingested mushrooms when visual identification is unreliable.

Amatoxin-Containing Mushroom Poisonings: Species, Toxidromes, Treatments, and Outcomes.

Amatoxins are produced primarily by 3 species of mushrooms: Amanita, Lepiota, and Galerina. Because amatoxin poisonings are increasing, the objective of this review was to identify all amatoxin-containing mushroom species, present a toxidromic approach to earlier diagnoses, and compare the efficacies and outcomes of therapies. To meet these objectives, Internet search engines were queried with keywords to select peer-reviewed scientific articles on amatoxin-containing mushroom poisoning and management. Descriptive epidemiological analyses have documented that most mushroom poisonings are caused by unknown mushrooms, and most fatal mushroom poisonings are caused by amatoxin-containing mushrooms. Amanita species cause more fatal mushroom poisonings than other amatoxin-containing species, such as Galerina and Lepiota. Amanita phalloides is responsible for most fatalities, followed by Amanita virosa and Amanita verna. The most frequently reported fatal Lepiota ingestions are due to Lepiota brunneoincarnata, and the most frequently reported fatal Galerina species ingestions are due to Galerina marginata. With the exception of liver transplantation, the current treatment strategies for amatoxin poisoning are all supportive and have not been subjected to rigorous efficacy testing in randomized controlled trials. All patients with symptoms of late-appearing gastrointestinal toxicity with or without false recovery or quiescent periods preceding acute liver insufficiency should be referred to centers providing liver transplantation. Patients with amatoxin-induced acute liver insufficiency that does not progress to liver failure will have a more favorable survival profile with supportive care than patients with amatoxin-induced acute liver failure, about half of whom will require liver transplantation.

A Case of Mushroom Poisoning with Russula subnigricans: Development of Rhabdomyolysis, Acute Kidney Injury, Cardiogenic Shock, and Death.

Mushroom exposures are increasing worldwide. The incidence and fatality of mushroom poisoning are reported to be increasing. Several new syndromes in mushroom poisoning have been described. Rhabdomyolytic mushroom poisoning is one of new syndromes. Russula subnigricans mushroom can cause delayed-onset rhabdomyolysis with acute kidney injury in the severely poisoned patient. There are few reports on the toxicity of R. subnigricans. This report represents the first record of R. subnigricans poisoning with rhabdomyolysis in Korea, describing a 51-year-old man who suffered from rhabdomyolysis, acute kidney injury, severe hypocalcemia, respiratory failure, ventricular tachycardia, cardiogenic shock, and death. Mushroom poisoning should be considered in the evaluation of rhabdomyolysis of unknown cause. Furthermore, R. subnigricans should be considered in the mushroom poisoning with rhabdomyolysis.

Russula subnigricans Poisoning: From Gastrointestinal Symptoms to Rhabdomyolysis.

Wild mushroom poisoning is often reported to cause acute liver or renal failure. However, acute rhabdomyolysis caused by wild mushroom poisoning has rarely been reported. We describe 7 patients of 1 family with Russula subnigricans Hongo poisoning. Their clinical manifestations varied from gastrointestinal symptoms to rhabdomyolysis, with 1 fatality. Our report provides supporting evidence that rhabdomyolysis may result from ingestion of R subnigricans mushrooms. A key to survival for patients with rhabdomyolysis caused by R subnigricans poisoning may be early recognition and intensive supportive care.

A Case Study: Rare Lepiota brunneoincarnata Poisoning.

Amatoxin poisoning from the genus Lepiota may have a deadly outcome, although this is not seen as often as it is from the genus Amanita. In this report, we present a patient who was poisoned by a sublethal dose of Lepiota brunneoincarnata mushrooms. The patient was hospitalized 12 hours after eating the mushrooms. The patient's transaminase levels increased dramatically starting on day 4. Aspartate transaminase peaked at 78 hours. Starting at 1265 IU/L, alanine transaminase peaked at 90 hours at 5124 IU/L. The patient was discharged on day 8 to outpatient care, and his transaminase levels returned to normal ranges in the subsequent days. A toxin analysis was carried out on the mushrooms that the patient claimed to have eaten. Using reversed-phase high-performance liquid chromatography analysis, an uptake of approximately 19.9 mg of amatoxin from nearly 30 g of mushrooms was calculated. This consisted of 10.59 mg of α-amanitin, 9.18 mg of ß-amanitin, and 0.16 mg of γ-amanitin. In conclusion, we present a patient from Turkey who was poisoned by L. brunneoincarnata mushrooms.

Intoxicaciones por setas, una actualización / Mushroom poisoning, an update

Las intoxicaciones por setas, motivadas por el consumo accidental de especies tóxicas, se producen todos los años durante la temporada de crecimiento de las mismas. Desde los motivos de la ingestión hasta los tratamientos médicos extremos como el trasplante hepático, pasando por la primera orientación diagnóstica, la derivación a urgencias y el ingreso en algunos casos, estas intoxicaciones son susceptibles de tener repercusiones médico-legales. Este trabajo ofrece una puesta al día sobre las intoxicaciones por setas que además de contribuir a mejorar su abordaje y tratamiento subraya diversos aspectos importantes desde la perspectiva médico-legal. Se describen diferentes aspectos clínicos, de diagnóstico y tratamiento de las principales intoxicaciones por setas, incluidas algunas descritas recientemente (AU)

Mushroom poisonings, caused by accidental intake of toxic species, occur every year during their growing season. From the cause of the ingestion to the application of extreme medical treatments such as liver transplantation, through the first diagnostic work up, referral to the emergency department and hospital admission in some cases, these poisonings are likely to have medico-legal implications.This paper provides an update on the subject of mushroom poisoning, which will help to improve their management and additionally highlight various important aspects from a forensic perspective. Diverse clinical, diagnostic and therapeutic matters of mushroom poisonings are depicted, including some new syndromes recently described (AU)

Laboratory confirmation of Amanita smithiana mushroom poisoning.

CONTEXT: Here we present a case of Amanita smithiana poisoning resulting in acute kidney injury requiring dialysis, and highlight laboratory methods used to confirm the diagnosis. Identification of Amanita smithiana toxin using thin-layer chromatography can provide greater diagnostic certainty than history and renal function tests alone. CASE DETAILS: A 63-year-old male presented to hospital with anuria and gastrointestinal symptoms, two days after consuming a soup of wild mushrooms he had picked. He was found to be in acute renal failure, requiring hemodialysis. After nine days of supportive treatment, he recovered renal function, and was discharged in good health 15 days post-ingestion. The patient provided a sample of leftover soup, and examination of cooked mushroom fragments by a mycologist provided preliminary identification of A. smithiana. Thin-layer chromatography revealed the presence of A. smithiana toxin in the soup, confirming this identification. DISCUSSION: A. smithiana is a nephrotoxic mushroom that can be easily mistaken for the edible and highly prized Pine mushroom (Tricholoma magnivelare). It causes initial gastrointestinal symptoms, followed by acute renal failure. Treatment includes dialysis and supportive care until the patient recovers renal function. The chemical structure of the A. smithiana toxin is unknown, but it can be identified as a characteristic spot on thin-layer chromatography.

Molecular detection of poisonous mushrooms in different matrices.

Amanita phalloides, Lepiota cristata, Lepiota brunneoincarnata and Inocybe asterospora are among the most important species responsible of mushroom poisoning in northern Italy. A real time PCR method for the identification of samples containing DNA from each of these species was developed. To test specificity all protocols were applied on DNA extracted from various mushroom species; sensitivity was assessed performing serial dilutions on all samples; versatility of the protocols was evaluated performing tests on DNA extracted from different matrices. The protocols showed high sensitivity (32 ng dried mushroom), high specificity and sensitive detection of DNA extracted from difficult samples, including pasta with mushroom, cooked mushrooms and gastric aspirates.

[Amanita pantherina poisoning or brain stroke?]. / Zatrucie muchomorem plamistym czy udar mózgu?

A case of a 72-year-old female who ingested panther cap (Amanita pantherina) was presented. During the ingestion of mushrooms she developed short-time diarrhea and severe transient neurological disorders; short-term hallucination followed by deep coma, skeletal muscle flaccidity with hyporeflexia. Also significant facial asymmetry was noted. A head CT showed a small ischaemic focus in capsula interna. After a neurological consult it was concluded that this small malformation was not the cause of severe patient's condition. The sporological test revealed numerous spores of panther cap in patient's vomit. After the next 6 hours complete resolution of neurological symptoms was observed. Finally it was assumed that patient's symptoms and signs were most probably a result of panther cap poisoning complicated with transient ischaemia of the brain.

[Foodborne infections and intoxications in Poland in 2003]. / Zatrucia i zakazenia pokarmowe w Polsce w 2003 roku.

A total of 20 221 bacterial foodborne infections and intoxications were registered in 2003 (incidence 52.9/100 000 population). These illnesses were less incident in 2003, compared both to 2002 figure (26 734 cases; incidence 69.9) and the median 1998-2001 (26 902 cases; incidence 69.6). Unlike bacterial infections, the incidence of viral foodborne infections increased in the recent years (2003--21.10 vs. 2002--17.30 and median 1998-2001 6.85). The parasitic foodborne infections did not change much during the recent years. A total of 78 cases of mushroom poisonings were reported (incidence 0.2) in 2003, compared to 66 cases in 2002 (incidence 0.17) and a median of 223 cases in 1998-2001 (incidence 0.6). The incidence of foodborne infections and intoxications was higher in the age group 0-4 years, compared to other age groups and in urban areas, compared to rural areas. A total of 230 foodborne and waterborne outbreaks involving 3816 cases were reported in 2003 (in Poland only outbreaks involving 4 persons or more were reported). The most prevalent etiological factors in outbreaks were Salmonella strains (63.6% of cases),Staphylococcus aureus (10.0% of cases) and Klebsiella strains (1.4% of cases). The most prevalent Salmonella strain were S. Enteritidis (89.3%) and S. Hadar (5.0%). The main vehicle of foodborne and waterborne outbreaks were meals prepared from various (> 2) raw materials of animal sources (29.2% of cases), egg meals (27.6%) and meats (7.2%). Of the places of food contamination, the most prevalent were own apartments (26.1% of outbreaks), food producing farms (10.6%), and restaurants (4.9%). One death was attributed to an outbreak of unknown etiology.

Management of maternal Amanita phalloïdes poisoning during the first trimester of pregnancy: a case report and review of the literature.

BACKGROUND: Amanita phalloïdes poisoning produces acute liver failure and often death. Maternal poisonings are rare, and medical decisions of abortion or liver transplantation in this critical situation frequently are based on laboratory data. We report here the case of a 22-year-old-woman in the 11th week of pregnancy, who ingested mushrooms. CASE REPORT: The patient's clinical symptoms (e.g., vomiting and diarrhea) and blood chemistry data (persistent increases of aspartate aminotransferase and alanine aminotransferase and severe decreases in prothrombin, factor V, factor II, factor VII, and factor X) indicated poisoning of medium severity. The management consisted of intravenous hydration, and administration of silymarine and N-acetylcysteine. No fetal damage was observed, and birth and development of the infant (now 2 years of age) proceeded without incident. CONCLUSION: Abortion is not necessarily indicated in maternal poisoning by A. phalloïdes, even in the first trimester of pregnancy.

A review of mushroom poisoning in Mexico.

A review of mycetisms reported in Mexico and a description of two interesting cases are given. Worldwide mycetisms are rarely studied. To identify the causal agents, clinical notes were analysed and field research with some of the persons involved in the poisonings were carried out. Symptoms and field data confirm a mortal hepatotoxic poisoning and a gastrointestinal mycetism produced by Amanita and Ramaria, respectively. In addition, mycetisms produced in Mexico by Boletus, Chlorophyllum, Conocybe, Hypomyces, Panaeolus, Psilocybe, Scleroderma, and Stropharia are discussed.

Multiple fatal mycetism caused by Amanita virosa in Mexico.

Mushroom poisonings caused by amatoxins are mostly lethal. Information about mycetisms caused by white species of Amanita is scarce. The present paper describes a case of mushroom poisoning caused by A. virosa. A prolongated latency period (6-10 hours), followed by cholera-like, improvement and visceral complication phases confirmed the amatoxin poisoning. The consumption of about 3 pounds of the toadstool by seven persons caused the death of five. Two patients survive the ingestion.

[The forensic medical diagnosis of death-cup poisonings]. / Sudebno-meditsinskaia diagnostika otravlenii blednoi pogankoi.

Comparative analysis and evaluation of all modern methods used for medicolegal diagnosis of poisoning with Amanita phalloides are presented. Imperfection of methods used for laboratory diagnosis of such poisonings requires extensive experimental control using the designed method of liquid chromatography for amanitine and phalloidine assay in the cadaveric organs and tissues.