Utilizing the DNA Aptamer to Determine Lethal α-Amanitin in Mushroom Samples and Urine by Magnetic Bead-ELISA (MELISA)

Amanita poisoning is one of the most deadly types of mushroom poisoning. α-Amanitin is the main lethal toxin in amanita, and the human-lethal dose is about 0.1 mg/kg. Most of the commonly used detection techniques for α-amanitin require expensive instruments. In this study, the α-amanitin aptamer was selected as the research object, and the stem-loop structure of the original aptamer was not damaged by truncating the redundant bases, in order to improve the affinity and specificity of the aptamer. The specificity and affinity of the truncated aptamers were determined using isothermal titration calorimetry (ITC) and gold nanoparticles (AuNPs), and the affinity and specificity of the aptamers decreased after truncation. Therefore, the original aptamer was selected to establish a simple and specific magnetic bead-based enzyme linked immunoassay (MELISA) method for α-amanitin. The detection limit was 0.369 μg/mL, while, in mushroom it was 0.372 μg/mL and in urine 0.337 μg/mL. Recovery studies were performed by spiking urine and mushroom samples with α-amanitin, and these confirmed the desirable accuracy and practical applicability of our method. The α-amanitin and aptamer recognition sites and binding pockets were investigated in an in vitro molecular docking environment, and the main binding bases of both were T3, G4, C5, T6, T7, C67, and A68. This study truncated the α-amanitin aptamer and proposes a method of detecting α-amanitin.

An Automated and Highly Sensitive Chemiluminescence Immunoassay for Diagnosing Mushroom Poisoning

Mushrooms containing Amanita peptide toxins are the major cause of mushroom poisoning, and lead to approximately 90% of deaths. Phallotoxins are the fastest toxin causing poisoning among Amanita peptide toxins. Thus, it is imperative to construct a highly sensitive quantification method for the rapid diagnosis of mushroom poisoning. In this study, we established a highly sensitive and automated magnetic bead (MB)-based chemiluminescence immunoassay (CLIA) for the early, rapid diagnosis of mushroom poisoning. The limits of detection (LODs) for phallotoxins were 0.010 ng/ml in human serum and 0.009 ng/ml in human urine. Recoveries ranged from 81.6 to 95.6% with a coefficient of variation <12.9%. Analysis of Amanita phalloides samples by the automated MB-based CLIA was in accordance with that of HPLC-MS/MS. The advantages the MB-based CLIA, high sensitivity, repeatability, and stability, were due to the use of MBs as immune carriers, chemiluminescence as a detection signal, and an integrated device to automate the whole process. Therefore, the proposed automated MB-based CLIA is a promising option for the early and rapid clinical diagnosis of mushroom poisoning.

Wild mushroom poisoning in Kumaon region of Uttarakhand: a case series

Mushrooms have been dietary source in hilly and ethnic tribes of India. More than 5000 mushroom species are known worldwide and nearly 100 species are known to be poisonous for humans. Mushroom poisoning occurs due to unintentional and accidental ingestion of poisonous mushroom due to misidentification of poisonous variety as edible one. There has been increasingly incidence of reporting of mushroom poisoning cases nowadays. Here we are reporting case series of 4 patients admitted hailing from same village with accidental ingestion of poisonous mushrooms with clinical-laboratory profile and outcome at our institution. Mushroom Poisoning is an emerging healthcare concern nowadays. Education and mass awareness for identification of poisonous mushrooms is an important preventive measure. Early hospitalization, proper hydration, gastric decontamination, silibinin and N- acetyl cysteine therapy with hepato-renal support constitutes mainstay of treatment. Delay in treatment and complications results in poor prognosis and mortality.

Two new Inosperma (Inocybaceae) species with unexpected muscarine contents from tropical China

An accurate identification of poisonous mushrooms and the confirmation of the toxins involved are both of great importance in the treatment of mushroom poisoning incidents. In recent years, cases of mushroom poisoning by Inosperma spp. have been repeatedly reported from tropical Asia. It is urgent to know the real species diversity of Inosperma in this region. In the present study, we proposed two new Inosperma species from tropical Asia, namely I. muscarium and I. hainanense. They were described based on morphology and multilocus phylogeny. Detailed descriptions, color photographs and the discussion with other closely related species of the two new taxa were provided. In addition, a comprehensive muscarine determination of these two new species using ultrahigh performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach has been performed. Results showed that these two species were muscarine positive, with a content of 16.03 ± 1.23 g/kg in I. muscarium and a content of 11.87 ± 3.02 g/kg in I. hainanense, much higher than the known species I. virosum. Recovery of muscarine ranged from 93.45% to 97.25%, and the average recovery is 95.56%.

COMPARATIVE INVESTIGATION OF KNOWLEDGE OF WILD EDIBLE MUSHROOMS IN VARNA REGION WITH OTHER FOREIGN GEOGRAPHIC ZONES

Purpose: The interest in evaluation of real knowledge of wild edible mushrooms at individual and population level increases mainly in developing countries. The purpose of our inquiry study was to assess the level of knowledge and awareness of the wild mushrooms in a random sample from the population of Varna region in Bulgaria. Material/Methods: We performed an anonymous questionnaire investigation of 200 adult individuals, 100 males and 100 females, from Varna region, including 17 potential opinions from their personal knowledge of wild edible mushroom usage. Statistical data processing was performed using descriptive methods, variation and correlation analyses. Results: Most respondents acquired their knowledge of wild edible mushrooms from their relatives. From all 200 persons investigated 78 respondents were not capable of recognizing at least one wild edible mushroom at all. More respondents trusted other people, who have very good experience with preparation of wild mushrooms, thus relying on differentiation of a mushroom poison. Their awareness was associated with their capacity to properly recognize wild mushrooms as well as their long-term practice to recognize toxic mushrooms. Just a minority of respondents shared that wild mushrooms in their vicinity could be a problem and had a slight idea of mycotherapy. Only two females had heard about training for wild mushroom recognition. There were statistically significant correlation dependences between respondent’s awareness of wild edible mushrooms, on one hand, and individual sex status, educational level and main residence, on the other hand. Conclusion: Our study identified two most recognized wild edible mushroom species in Varna region, edible boletus (Boletus edulis Bull) and parasol mushroom (Macrolepiota procera S. F. Gray). Most respondents acquired their knowledge of wild edible mushrooms from relatives. A problem-oriented training of the population on this topic should be performed to improve awareness of common mushroom features and avoid mushroom poisoning.

Acute liver failure caused by Amanita verna: a case series and review of the literature

Background Amanita verna is one of the most harmful wild fungi in China. Amanita verna poisoning occurs every year, and the mortality is as high as 50%. However, its clinical manifestations are complex and diverse. Case presentation In March 2019, three patients took a large amount of Amanita, and one of them received liver transplantation in Zhongshan hospital, Sun Yat-sen University. All patients had vomiting and diarrhea 8–12 h after eating wild mushrooms (Amanita). The patients were initially diagnosed with Amanita poisoning. One case (case 3) was complicated and diagnosed as mushroom poisoning (fatal Amanita), toxic hepatitis, acute liver failure, toxic encephalopathy, hemorrhagic colitis, toxic myocarditis, disseminated intravascular coagulation (DIC) and pregnancy. The general clinical data of all patients were recorded, who received early treatment such as hemodialysis, artificial liver plasma exchange, hormone shock and anti-infection. One case (case 1) recovered smoothly after liver transplantation, and the indexes of liver, kidney, coagulation function and infection were improved. The other two cases died of intracerebral hemorrhage. Conclusion Liver transplantation is an effective method for the treatment of acute liver failure caused by mushroom poisoning and can improve the survival rate of patients with toxic liver failure.