[Study on 4 cases of mushroom poisoning with amanita and identification of poison].

Different kinds of poisonous mushrooms contain different toxic components. Acute liver injury caused by amanita mushroom is the main cause of death from poisonous mushroom poisoning in China. Consumption of poisonous mushrooms has an incubation period, there is a false recovery period in the clinical process, and the early performance is slight and does not attract enough attention from doctors, and it is easy to miss the treatment opportunity. The clinical characteristics, treatment and identification of mushrooms containing amanita in 4 patients were analyzed in order to improve clinicians' understanding of the diagnosis and treatment of mushroom poisoning and early species identification.

Rapid Russula senecis identification assays using loop-mediated isothermal amplification based on real-time fluorescence and visualization.

Russula senecis, a common poisonous mushroom, is widely distributed in China. Mushroom poisoning is becoming a major threat to human health and its rate is increasing worldwide. For the first time, we developed a set of loop-mediated isothermal amplification (LAMP) assays based on a real-time fluorescence and a visualization method to detect R. senecis, and the visual LAMP reaction system was optimized to further shorten the reaction time. Both real-time LAMP and visual LAMP could detect as low as 3.2 pg of genomic DNA. In addition, fried and digested mushrooms were used to validate the proposed LAMP method, and mushroom mixtures with as low as 1% of the target species could be successfully detected, indicating that the LAMP assays established in this study had good applicability and could be used for clinical sample detection and forensic identification. Furthermore, the LAMP assays were proven to be comparable to the real-time PCR method. KEY POINTS: • A set of loop-mediated isothermal amplification (LAMP) assays based on real-time fluorescence and visualization to detect Russula senecis was developed. • Both real-time LAMP and visual LAMP can be used to detect genomic DNA at concentrations as low as 3.2 pg. • By simulating mushroom processing and digestion in gastric juice, LAMP assays were proved to have good applicability and could be used for clinical diagnosis and forensic analysis.

Surveillance for foodborne disease outbreaks in Zhejiang Province, China, 2015-2020.

BACKGROUND: Foodborne diseases are a major cause of morbidity and mortality and a major public health problem worldwide. We aim to explore characteristics of foodborne disease outbreaks (FBDOs) in Zhejiang Province and to provide data support for foodborne disease prevention and control. To our knowledge, few such analyses have been published at the provincial level. METHODS: Descriptive statistical methods were used to analyze the data reported by centers for disease control (CDC) at all levels in Zhejiang Province through Foodborne Disease Outbreaks Surveillance System (FDOSS) during 2015-2020. RESULTS: A total of 962 FBDOs were reported during this period, resulting in 8324 illnesses, 1028 hospitalizations, and 20 deaths. The number of outbreaks (410 outbreaks, 42.62%) and cases (4991 cases, 59.96%) caused by bacteria were the largest, followed by poisonous mushrooms (157 outbreaks, 587 cases), which was the main cause of death (15 deaths, 75%). The highest number of FBDOs occurred in households (381 outbreaks, 39.60%), followed by restaurants (219 outbreaks, 22.77%) and canteens (174 outbreaks, 18.08%). Animal-based foods were the most common single food reported (232 outbreaks, 24.12%), followed by poisonous mushrooms (162 outbreaks, 16.84%), and plant-based foods (133 outbreaks, 13.83%). Poisonous mushrooms took the first place in outbreaks in households (38.32%, 146/381), while bacteria took the first place in outbreaks outside households. Vibrio parahaemolyticus was responsible for the largest number of outbreaks (232 outbreaks, 24.12%), which mainly occurred in catering service units (93.10%, 216/232). Different types of bacteria tended to be found in different food categories, such as Vibrio parahaemolyticus, which was mainly found in aquatic products. CONCLUSIONS: Analysis of FBDOs can provide insight into the most important pathogens and sources of foodborne disease, helping authorities identify high-risk etiologies, high-risk foods, and high-risk settings to guide policies that would reduce FBDOs.

[Three cases of mushroom poisoning caused by lethal Amanita species].

Mushroom poisoning with amatoxins can cause liver dysfunction in patients, and death in severe cases. The amatoxins detection by enzyme-linked immunosorbent assay (ELISA) can help early clinical diagnosis. Three patients were identified as α-amatoxin containing mushroom poisoning by ELISA. The first symptoms of patients was gastrointestinal symptoms, and liver function damage occured later. One patient gave up treatment and died. After received supportive treatments such as adsorption of toxins, catharsis, fluid supplementation to promote toxin metabolism and liver protection, 2 patients were recovered and discharged.

[An incident of toadstool poisoning].

Objective: To investigate a poisonous mushroom poisoning incident and analyze its clinical data. Methods: Investigate a poisonous mushroom poisoning incident in a place in Shandong in July 2018, at the same time, the clinical data of 2 cases of mushroom poisoning were analyzed and summarized. Results: The incident was caused by a poisoning incident caused by residents eating poisonous mushrooms. The poisonous mushroom in this incident was identified as a scaled white goose cream. Two patients with poisoning developed gastrointestinal symptoms such as nausea and vomiting, abdominal pain and diarrhea, and liver damage occurred later. After active rescue and treatment, one patient was discharged from hospital, and the other patient developed acute pulmonary embolism during the treatment. He was discharged after interventional thrombolysis and follow-up treatment. Conclusion: After investigation, the incident was caused by the ingestion of poisonous mushrooms mainly based on the scalloped white goose cream. After active treatment, they were cured and discharged.

Expansion and diversification of the MSDIN family of cyclic peptide genes in the poisonous agarics Amanita phalloides and A. bisporigera.

BACKGROUND: The cyclic peptide toxins of Amanita mushrooms, such as α-amanitin and phalloidin, are encoded by the "MSDIN" gene family and ribosomally biosynthesized. Based on partial genome sequence and PCR analysis, some members of the MSDIN family were previously identified in Amanita bisporigera, and several other members are known from other species of Amanita. However, the complete complement in any one species, and hence the genetic capacity for these fungi to make cyclic peptides, remains unknown. RESULTS: Draft genome sequences of two cyclic peptide-producing mushrooms, the "Death Cap" A. phalloides and the "Destroying Angel" A. bisporigera, were obtained. Each species has ~30 MSDIN genes, most of which are predicted to encode unknown cyclic peptides. Some MSDIN genes were duplicated in one or the other species, but only three were common to both species. A gene encoding cycloamanide B, a previously described nontoxic cyclic heptapeptide, was also present in A. phalloides, but genes for antamanide and cycloamanides A, C, and D were not. In A. bisporigera, RNA expression was observed for 20 of the MSDIN family members. Based on their predicted sequences, novel cyclic peptides were searched for by LC/MS/MS in extracts of A. phalloides. The presence of two cyclic peptides, named cycloamanides E and F with structures cyclo(SFFFPVP) and cyclo(IVGILGLP), was thereby demonstrated. Of the MSDIN genes reported earlier from another specimen of A. bisporigera, 9 of 14 were not found in the current genome assembly. Differences between previous and current results for the complement of MSDIN genes and cyclic peptides in the two fungi probably represents natural variation among geographically dispersed isolates of A. phalloides and among the members of the poorly defined A. bisporigera species complex. Both A. phalloides and A. bisporigera contain two prolyl oligopeptidase genes, one of which (POPB) is probably dedicated to cyclic peptide biosynthesis as it is in Galerina marginata. CONCLUSION: The MSDIN gene family has expanded and diverged rapidly in Amanita section Phalloideae. Together, A. bisporigera and A. phalloides are predicted to have the capacity to make more than 50 cyclic hexa-, hepta-, octa-, nona- and decapeptides.

Two Photosensitive Dermatitis Outbreaks Caused by Cordierites frondosus - Chuxiong Yi Autonomous Prefecture, Yunnan Province, China, 2023.

What is already known about this topic?: Cordierites frondosus (C. frondosus) is a species of toxic mushroom known to induce symptoms of photosensitive dermatitis. What is added by this report?: In the months of May and June 2023, a total of four patients in Chuxiong Yi Autonomous Prefecture, Yunnan Province, were affected by C. frondosus poisoning, occurring over two distinct incidents. The condition of two patients deteriorated after they were re-exposed to sunlight on the seventh day following the initial poisoning. Separately, an additional two patients reported experiencing a mild, needle-like sensation on areas of their skin exposed to the sun, recorded on the twelfth day subsequent to the poisoning. What are the implications for public health practice?: Given that symptoms of photosensitive dermatitis, a potential severe consequence of C. frondosus poisoning, can manifest up to a week post-sun exposure, it is advisable to avoid sunlight for a minimum of two weeks following poisoning.

Mushroom Poisoning Outbreaks - China, 2023.

What is already known about this topic?: Mushroom poisoning poses a significant food safety concern in China, with a total of 196 species identified in poisoning incidents by the end of 2022. What is added by this report?: In 2023, the China CDC conducted an investigation into 505 cases of mushroom poisoning spanning 24 provincial-level administrative divisions. This investigation resulted in 1,303 patients and 16 deaths, yielding a case fatality rate of 1.23%. A total of 97 mushrooms were identified as the cause of 6 distinct clinical disease types, with 12 species newly documented as poisonous mushrooms in China. What are the implications for public health practice?: Close collaboration among CDC staff, physicians, and mycologists remains crucial for the control and prevention of mushroom poisoning in the future.

Electrochemical biosensor based on the integration of maple leaf-like gold nanocrystal and truncated aptamer for detection of α-amanitin with high sensitivity, selectivity and rapidity.

This study reports the fabrication of three-dimensional gold nanocrystals as sensing material in the presence of l-glutathion and high-performance aptamer with 20 bases of α-amanitin via truncation and optimization of along aptamer. The resulting maple leaf-like gold nanocrystal (ML-Au) exhibits an improved catalytic activity due to more exposed high-index facets. The use of truncated aptamer increases the sensitivity by 15 times and reduces the reaction time by two times compared with those of original aptamer. An α-amanitin electrochemical biosensor constructed by integrating ML-Au nanocrystals with truncated aptamer exhibits high sensitivity, selectivity and rapidity. An increase of the α-amanitin concentration in the range of 1 × 10-14-1 × 10-9 M causes a linear decrease in the amperometric current with a limit of detection of 2.9 × 10-15 M (S/N = 3). The proposed analytical method is satisfactorily used for electrochemical sensing of α-amanitin in urine and wild mushroom samples.

Polymerase chain reaction-based methods for the rapid identification of Amanita exitialis.

Amanita exitialis, a deadly mushroom found in eastern Asia, causes the highest death rates among all poisonous mushrooms in China. The aim of the present study was to develop an efficient, accurate, and user-friendly PCR-based method for identifying A. exitialis that could facilitate the prevention, diagnosis, and treatment of associated food poisoning. A. exitialis-specific primers and probes were designed based on the internal transcribed spacer region variations of 27 mushroom species. Specificity was confirmed using conventional and real-time PCR for 23 non-target mushroom species, including morphologically similar and closely related species. Compared to conventional PCR, real-time PCR was more sensitive (detectable DNA concentration: 1.36 × 10-2 ng/µL vs. 1.36 × 10-3) and efficient (analysis time: 1 h vs. 40 min). Furthermore, the real-time PCR results could be immediately visualized using amplification curve analysis. The results present two robust PCR-based methods for A. exitialis identification that can facilitate food safety.

Mushroom Poisoning Outbreaks - China, 2022.

What is already known about this topic?: Mushroom poisoning is one of the most serious food safety issues in China. By the end of 2021, over 520 poisonous mushrooms had been discovered in China. The Southwest region of China was the most severely affected. Mushroom poisonings mainly concentrated in the summer and autumn months. What is added by this report?: In 2022, China CDC conducted an investigation of 482 incidents of mushroom poisoning across 21 provincial-level administrative divisions (PLADs). This resulted in 1,332 patients and 28 deaths, with a total case fatality rate of 2.1%. A total of 98 mushrooms were identified, causing 7 different clinical types of diseases. Three provisional new species (Collybia humida nom. prov., Spodocybe venenata nom. prov., and Omphalotus yunnanensis nom. prov.) were newly recorded as poisonous mushrooms in China, in addition to 10 other species. What are the implications for public health practice?: In view of the extensive impact and harm of poisonous mushrooms on public health, it is necessary to promote prevention and improve the ability of professionals to identify, diagnose, and treat mushroom poisoning.

[Determination of five amanita peptide toxins in poisonous mushrooms by ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high resolution mass spectrometry].

Food poisoning by toxic mushrooms occurs frequently worldwide. It is one of the most common food poisoning events and the main cause of death. Amanita peptide toxins are the most common lethal toxins in poisonous mushrooms. Presently, a novel method based on ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q/Orbitrap HRMS) was developed for the determination of five amanitapeptide toxins (α-amanitin, ß-amanitin, γ-amanitin, phalloidin, and phallacidin). Because the isotope summit of α-amanitin affects the detection of ß-amanitin, it cannot be distinguished by low resolution mass spectrometry. Therefore, experimental conditions including chromatography and mass spectrometry were explored in detail. The five peptide toxins were extracted from poisonous mushrooms with pure water and filtered through a 0.22 µm teflon microporous membrane. The procedure was rapid, simple, and environmentally friendly. Chromatographic separation was performed on a strong polarity HSS T3 column (100 mm×2.0 mm, 2.1 µm) with gradient elution using acetonitrile and 5 mmol/L ammonium acetate containing 0.1% (v/v) formic acid as mobile phases at a flow rate of 0.3 mL/min. The column temperature was set to 40 ℃. The analytes were ionized using a heating electrospray ionization source and collected in positive ion mode. Full scanning/data-dependent secondary mass spectrometry (Full mass-ddMS2) mode was used for qualitative analysis of the targets within 10 min. The target ion selective scan (Targeted-SIM) mode was used for quantification by external standard calibration. The measured and theoretical values of the exact mass and the MS2 fragment ions of the five compounds were within an error of 5×10-6. Method validation was performed according to the criteria recommended by the Chinese National Standard. All the compounds showed an excellent linear relationship in the range of 1.0-20.0 µg/L. The correlation coefficients (r) ranged from 0.9974 to 0.9989. The limit of detection was 0.006 mg/kg for all five compounds. Recoveries ranged from 81.8% to 102.4%. There was no matrix effect in the blank mushroom sample for the five compounds, and the relative standard deviations ranged from 3.2% to 8.3%. This method provides abundant compound characteristic mass information, such as retention time, exact mass, fragment ions, and other information. The data can be used to identify suspected compounds based on the extracted ion flow diagram and isotope distribution information. Comparison between the actual exact mass and the theoretical exact mass, combined with the fragment ions enables identification of the structures of unknown compounds and collision methods, which can be confirmed in the absence of standard materials. In this study, the isomer of γ-amanitin was identified as amaninamide. The novel method is simple, accurate, specific, and sensitive. The method permits the rapid qualitative and quantitative detection of compound in public health emergency settings and will provide reliable technical support for the rapid screening of such toxic compounds and the structural locking of unknown toxins in the future.

Mushroom poisoning in Thailand between 2003 and 2017.

The frequent occurrence of mushroom poisoning cases in the wet season in Thailand has long been recognized but has never been quantitatively analyzed. This study aims to analyze mushroom poisoning cases in Thailand between 2003 and 2017 and focused on their association with the rainfall. The results revealed 22,571 cases and 106 deaths in this period. Cases were higher for females than males, adults than children, and agriculturists than people in other occupations. Cases were higher in the northeastern and northern regions than in the central, south, and east regions. There are strong effects of seasonality on mushroom poisoning cases. Over the 15-y period, 17,337 cases occurred in the wet season (May-Sep) compared with 5,234 cases in the dry season (Oct-Apr). The number of monthly poisoning cases were high in men, mature adults, agriculturists, people living in the rural areas and people living in the Northeastern and Northern provinces. Moreover, a strong positive correlation was shown between the number of monthly poisoning cases and the amount of monthly rainfall.

Mushroom Poisoning Outbreaks - China, 2021.

What is already known about this topic?: Mushroom poisoning is one of the most serious food safety issues in China. Most poisoning incidents resulted from eating mushrooms causing gastroenteritis and psycho-neurological disorder from which patients usually could fully recover. Most deaths resulted from species causing acute liver failure and rhabdomyolysis, and the remaining deaths were attributed to acute renal failure and hemolysis. What is added by this report?: In 2021, the total number of investigations was 327 from 25 provincial-level administrative divisions, involving 923 patients and 20 deaths, and the overall mortality was 2.17%. Overall, 74 poisonous mushrooms causing 6 different clinical syndromes were successfully identified, 15 of which were newly recorded in China as poisonous mushrooms. What are the implications for public health practice?: Considering the potential huge risks for collecting and eating wild mushrooms, we strongly advise not collecting and eating unfamiliar wild mushrooms. Promoting knowledge about poisonous mushrooms is essential and urgent to reduce mushroom poisonings. Precise species identification timely after mushroom poisoning is important for appropriate diagnosis and treatment. Many deaths were ascribed to delayed hospitalization.

Mushroom mycetism - A neglected and challenging medical emergency in the Indian subcontinent: A road map for its prevention and treatment.

Edible mushrooms, a class of macroscopic fungi, serve as delicious and nutritious food supplements around the world. Nevertheless, accidental consumption of poisonous mushrooms that results in fatality or severe illness is typical in all countries, especially among the tribal indigenous communities that forage wild mushrooms for food. In the Indian subcontinent, mushroom poisoning cases are underreported and neglected. Different classes of toxins, characterized from the poisonous mushrooms found globally, show variable clinical symptoms post-consumption. Although the Indian subcontinent is a biodiversity hotspot and home to different classes of fungi and mushrooms, many species of poisonous mushrooms and their toxins, have yet to be identified and characterized. No epidemiological studies or retrospective analyses of mushroom poisoning cases have been reported from the poison control centers in the Indian subcontinent. Nevertheless, some limited clinical and epidemiological data is available from India and Nepal, and therefore, we critically analyse the mushroom poisoning scenario in these countries, and discuss the mushroom toxins that are likely responsible for the post-ingestion toxicities. We also correlate the clinical manifestations of mushroom intoxication in India and Nepal with the pharmacological properties of the prevalent mushroom toxins in these countries. Our limited study of mushroom poisoning demonstrates that the adverse pharmacological effects of amatoxin, one of the deadliest mushroom toxins, are responsible for the highest mortality and morbidity in India and Nepal. Further, no specific antidote is available to treat mushroom intoxication in the region, and systemic and supportive care is all that is available for in-patient management of cases of severe poisoning. We also suggest a roadmap for the prevention and specific treatment against mushroom poisoning in the Indian subcontinent.

The complete mitochondrial genome of the poisonous mushroom Trichoderma cornu-damae (Hypocreaceae).

Trichoderma cornu-damae is a poisonous mushroom that contains trichothecene mycotoxins. The complete mitochondrial genome of this mushroom was determined using next-generation sequencing. This mitogenome is a circular molecule 94,608 bp in length with a GC content of 27.94% and contains 15 protein-coding genes, two rRNA genes (rnl and rns), and 25 tRNA genes. Phylogenetic analysis placed T. cornu-damae in the family Hypocreaceae group, which includes the genus Trichoderma. The mitogenome of T. cornu-damae will contribute to our understanding of the phylogeny, taxonomy, and population genetics of this mushroom.

Draft Genomes of Six Wild Poisonous Mushrooms.

Foodborne illnesses caused by wild mushroom poisoning occur globally and have led to food safety concerns. Here, we reported de novo genome assemblies of the six most commonly encountered toxic mushrooms in Thailand. These comprised Amanita brunneitoxicaria, Cantharocybe virosa, Chlorophyllum molybdites, Entoloma mastoideum, Pseudosperma sp. and Russula subnigricans. The nuclear genome sizes of these species ranged from 40 to 77 Mb, with the number of predicted genes ranging from 5,375 to 14,099. The mitogenome sizes varied from 41,555 to 78,907 bp. The resulting draft genomes of these poisonous mushrooms provide insights into toxin-related genes that may be used to establish genetic markers for monitoring mushroom poisoning outbreaks.

 Pseudospermaarenarium (Inocybaceae), a new poisonous species from Eurasia, based on morphological, ecological, molecular and biochemical evidence.

In this study, Pseudospermaarenarium is proposed as a new species, based on morphological, ecological, molecular and biochemical evidence. The new species grows on sandy ground under Populus and Pinussylvestris in north-western China and northern Europe, respectively. It is characterised by the combination of the robust habit, nearly glabrous pileus, large cylindrical basidiospores, thin-walled cheilocystidia and ecological associations with Populusalba × P.berolinensis and Pinussylvestris and unique phylogenetic placement. Additionally, a comprehensive toxin determination of the new species using ultra-high performance liquid chromatography-tandem mass spectrometry was conducted. Results showed that it was a muscarine-positive species. The content were approximately five times higher in the pilei [4012.2 ± 803.1-4302.3 ± 863.2 mg/kg (k = 2, p = 95%)] than in the stipes [850.4 ± 171.1-929.1 ± 184.2 mg/kg (k = 2, p = 95%)], demonstrating the severity of mushroom poisoning when patients consumed different parts of the poisonous mushroom. Amatoxins, phallotoxins, ibotenic acid, muscimol, psilocybin and psilocin were not detected.

Toxin screening of Pseudosperma umbrinellum (Agaricals, Basidiomycota): First report of phalloidin in Inocybaceae mushroom.

Pseudosperma species are widely distributed worldwide. Many of them cause poisoning incidents every year, and the toxin responsible for poisoning is muscarine, which could stimulate the parasympathetic nervous system. This study established a method using multiwalled carbon nanotube purification and liquid chromatography-tandem mass spectrometry for the targeted screening of mushroom toxins (muscarine, isoxazole derivatives, tryptamine alkaloids, three amatoxins and three phallotoxins) from Pseudosperma umbrinellum, a common poisonous mushroom distributed in north and northwestern China. Surprisingly, in addition to muscarine, phalloidin was also detected in P. umbrinellum, and the contents were 3022.2 ± 604.4 to 4002.3 ± 804.6 mg/kg (k = 2; p = 95%) muscarine and 5.9 ± 1.2 to 9.3 ± 1.8 mg/kg (k = 2; p = 95%) phalloidin.

Insight into isolation and elucidation of cytotoxic ergostanoids from the mushroom Sarcosphaera crassa (Santi) Pouzar: An edible mushroom.

Sarcosphaera crassa is a mushroom consumed in Europe and Anatolia after being cooked well. The cytotoxic activity of the extracts of unbaked S. crassa against MCF7, HT29, HeLa cancer cell lines and toxicity against PDF fibroblast healthy cell lines were studied using MTT assay. Acetone and methanol extracts of the mushroom exhibited significant cytotoxic activity. Further investigation of cytotoxic extracts afforded two new fatty acid sterols (1-2), a new ergosterol glycoside (4), and seven known compounds, including a fatty acid sterol (3), a steroid glycoside (5), two ergostanoids (6-7) and three sugars (8-10). These compounds were identified as brassicasteryl heptadecanoate (1), brassicasteryl palmitoleate (2), brassicasteryl linoleate (3), brassicasterol ß-ᴅ-xylofuranoside (4), brassicasterol ß-ᴅ-glucoside (5), brassicasterol (6), ergosterol-endoperoxide (7), mannitol (8), erythritol (9) and turanose (10). Among them, 7 exhibited a moderate cytotoxic activity against HeLa (IC50: 70.1 ± 2.0 µg/mL) and high activity against HT29 (IC50: 38.8 ± 0.9 µg/mL), and MCF7 (IC50: 62.9 ± 1.3 µg/mL) cancer cell lines. Compounds 4, 5, and 6 also exhibited significant cytotoxic activity against HT29 and MCF7. Moreover, all compounds exhibited weak toxicity against PDF healthy cell lines. This study indicates the potential use of Sarcosphaera crassa as a natural source of cytotoxic ergostanoids, which can be considered a dietary supplement for cancer prevention.