DOCSLIB.ORG

Quantification of Alpha-Amanitin in Biological Samples by HPLC Using Simultaneous UV

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Quantification of Alpha-Amanitin in Biological Samples by HPLC Using Simultaneous UV Journal of Chromatography B, 997 (2015) 85–95 View metadata, citation and similar papers at core.ac.uk brought to you by CORE Contents lists available at ScienceDirect provided by Biblioteca Digital do IPB Journal of Chromatography B journal homepage: www.elsevier.com/locate/chromb Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection Juliana Garcia a,∗, Vera M. Costa a, Paula Baptista b, Maria de Lourdes Bastos a, Félix Carvalho a,∗ a REQUIMTE/Laboratory of Toxicology, Department of Biological Sciences, Faculty of Pharmacy, University of Porto, Rua José Viterbo Ferreira n◦ 228, 4050-313 Porto, Portugal b CIMO/School of Agriculture, Polytechnic Institute of Braganc¸ a, Campus de Santa Apolónia, Apartado 1172, 5301-854 Braganc¸ a, Portugal article info abstract Article history: ␣-Amanitin is a natural bicyclic octapeptide, from the family of amatoxins, present in the deadly mush- Received 6 November 2014 room species Amanita phalloides. The toxicological and clinical interests raised by this toxin, require Received in revised form 27 May 2015 highly sensitive, accurate and reproducible quantification methods for pharmacokinetic studies. In the Accepted 4 June 2015 present work, a high-performance liquid chromatographic (HPLC) method with in-line connected diode- Available online 10 June 2015 array (DAD) and electrochemical (EC) detection was developed and validated to quantify ␣-amanitin in biological samples (namely liver and kidney). Sample pre-treatment consisted of a simple and unique Keywords: deproteinization step with 5% perchloric acid followed by centrifugation at 16,000 × g,4◦C, for 20 min. High-performance liquid chromatographic ␣ ≥ ␣ Diode-array The high recovery found for -amanitin ( 96.8%) makes this procedure suitable for extracting -amanitin Electrochemical from liver and kidney homogenates. The resulting supernatant was collected and injected into the HPLC. ␣-amanitin Mobile phase was composed by 20% methanol in 50 mM citric acid, and 0.46 mM octanessulfonic acid, adjusted to pH 5.5. The chromatographic runs took less than 22 min and no significant endogenous inter- ferences were observed at the ␣-amanitin retention time. Calibration curves were linear with regression coefficients higher than 0.994. The overall inter- and intra-assay precision did not exceed 15.3%. The present method has low interferences with simple and fast processing steps, being a suitable procedure to support in vivo toxicokinetic studies involving ␣-amanitin. In fact, the validated method was successfully applied to quantify ␣-amanitin in biological samples following intraperitoneal ␣-amanitin administration to rats. Moreover, human plasma was also used as matrix and the purposed method was adequate for detection of ␣-amanitin in that matrix. The results clearly indicate that the proposed method is suitable to investigate the pharmacokinetic and tissue distribution of ␣-amanitin. Additionally, the method will be very useful in the development of novel and potent antidotes against amatoxins poisoning and to improve the knowledge of ␣-amanitin toxicity. © 2015 Elsevier B.V. All rights reserved. 1. Introduction in this mushroom and is the main responsible for its toxic effects [1]. ␣-Amanitin produces hepatic and kidney failure, which result The main toxin of the amatoxins group, ␣-amanitin (Fig. 1)isa in a potentially fatal outcome in most of the human intoxication toxic bicyclic octapeptide present in the deadly mushroom species cases [2]. The underlying toxicity mechanisms of ␣-amanitin are Amanita phalloides. It represents up to 40% of the amatoxins present not completely understood, but it is well accepted that the inhi- bition of RNA polymerase II (RNAP II) is its main effect [3] . Other mechanisms also appear to be involved, including oxidative stress [4]. Abbreviations: HPLC, high-performance liquid chromatography; DAD, diode- So far, there is no specific and totally effective antidote array; EC, electrochemical; DNA, deoxyribonucleic acid; LOD, limit of detection; against A. phalloides poisoning. Benzylpenicillin, ceftazidime, N- LOQ, limit of quantification; LC, liquid chromatography; MS, mass spectrometry; acetylcysteine, and silybin are the most extensively used antidotes, RNA, ribonucleic acid; RNAP II, RNA polymerase II; RSD, relative standard deviation; SPE, solid phase extraction; UV, ultraviolet. however mortality remains high [5]. Currently, in vivo and in silico ∗ Corresponding authors. Fax: +351 22609339. studies are ongoing in search of effective antidotes, either directed E-mail addresses: jugarcia [email protected] (J. Garcia), [email protected] to the toxicokinetics or to the toxicodynamics of amatoxins [6–9] (F. Carvalho). http://dx.doi.org/10.1016/j.jchromb.2015.06.001 1570-0232/© 2015 Elsevier B.V. All rights reserved. 86 J. Garcia et al. / J. Chromatogr. B 997 (2015) 85–95 Fig. 1. Chemical structure of ␣-amanitin. In both approaches, it is expected that the development of more with a Milli-Q Plus ultra-pure water purification system (Millipore, insightful analytical and physiological methodologies will provide Bedford, Massachusetts, USA). essential tools for achieving this purpose. ␣ Besides the toxicological interest in -amanitin, recent findings 2.2. Chromatographic conditions indicate that ␣-amanitin is emerging as an attractive new class of potent anticancer drug [10]. For this clinical application, ade- HPLC analysis was performed using a Waters 2690 separa- quate analytical methodologies are also required for the evaluation tion module (Waters, Milford, MA, USA), and a 250 mm × 4.6 mm, ␣ of -amanitin pharmacokinetics, both in pre-clinical and clinical Waters Spherisorb RP-18 (5 ␮m) ODS2 column. The mobile phase studies. (20% methanol in 50 mM citric acid, 0.46 mM octanessulfonic acid, The comprehensive characterization of antidotes in A. phal- adjusted to pH 5.5 with 10 M NaOH) was filtered through a 0.45 ␮m loides poisoning and the pharmacokinetics and toxicokinetic membrane (Millipore, Madrid, Spain) and degassed. An isocratic ␣ studies of -amanitin require highly sensitive, accurate and elution was performed at a flow rate of 1.0 mL/min, at room temper- reproducible quantification assays. Currently, there are several ature. Sample aliquots of 20 ␮L were injected. A Compaq computer high-performance liquid chromatography (HPLC)–ultraviolet (UV) fitted with Millenium32 software (Waters) processed the chro- [11–17], liquid chromatography (LC)–mass spectrometry (MS) matographic and spectral data. [18–20] and ultra performance liquid chromatography (UPLC)- For the spectroscopic and coulochemical analysis, a photodi- ␣ MS/MS [21] methods to determine -amanitin in mushrooms, liver ode array detector (Waters model 996) and/or a Colouchem II ␣ and biological fluids (Table 1). HPLC-MS methods for -amanitin (ESA, Chelmsford, USA) equipped with a guard cell (ESA 5020) and detection are accurate, sensitive, and specific; nevertheless HPLC- analytical cell (ESA 5011A) electrochemical detector were used. MS is an expensive technique and is not available in many In all the studies, both detectors were used, with the analytical laboratories. To date there are no published HPLC with electro- cell of the coulochemical detector placed after the photodiode ␣ chemical (EC) detection methods for the analysis of -amanitin in array detector. The electrochemical potential settings of Coulochem biological tissues. Hence, in this paper, we describe a simple acidic II detector were: guard cell, −550 mV, and analytical detector + extraction and HPLC methodology with in-line connected photodi- 500 mV, following a previous reported procedure [22]. A current ␣ ode array detection (DAD) and EC detection to quantify -amanitin of 5 ␮A full-scale was used. Ultraviolet-visible spectra were col- in rat liver and kidney. Moreover, we applied the validated method lected for all samples between from 230 to 400 nm. For quantitative ␣ to identify -amanitin in human plasma. The validated method analyses, chromatograms were integrated at 305 nm [15]. was successfully applied to quantify ␣-amanitin in plasma, liver and kidney tissues following intraperitoneal administration to rats. The new method provides the required high analytical sensitivity 2.3. Stock solution and calibration standards and reproducibility, with a simple and fast pretreatment, as it is ␣ required for advanced pharmacokinetic and toxicokinetic research. A stock solution of -amanitin was prepared in water Milli-Q and diluted to a final concentration of 1 mg mL−1. All stock solu- tions were stored of −20 ◦C. A set of eleven standards ranging from 0.025 ␮gmL−1 to 10 ␮gmL−1 were prepared at the moment of 2. Material and methods use by spiking the rat liver and kidney samples with appropriate amounts of ␣-amanitin. All calibration standards were prepared in 2.1. Reagents and chemicals triplicate. ␣-Amanitin (>99%), bovine serum albumin, and 1- 2.4. Animals and sample preparation octanesulfonic acid sodium salt were purchased from Sigma-Aldrich (St. Louis, MO, USA). Analytical HPLC grade The liver and kidney tissues were obtained from adult male methanol was obtained from Fisher Scientific (Waltham, Mas- Wistar rats. These animals were used as healthy controls in other sachusetts, USA). Sodium chloride (NaCl), potassium chloride experiments, where liver and kidney were not necessary. A mini- (KCl), sodium hydrogen phosphate (Na2HPO4), potassium dihy- mum number of animals was used in order to obtain valid results. drogen phosphate
Load more

Recommended publications
  • Four Patients with Amanita Phalloides Poisoning CASE SERIE
    CASE SERIE 353 Four patients with Amanita Phalloides poisoning S. Vanooteghem1, J. Arts2, S. Decock2, P. Pieraerts3, W. Meersseman4, C. Verslype1, Ph. Van Hootegem2 (1) Department of hepatology, UZ Leuven ; (2) Department of gastroenterology, AZ St-Lucas, Brugge ; (3) General Practitioner, Zedelgem ; (4) Department of internal medicine, UZ Leuven. Abstract because they developed stage 2 hepatic encephalopathy. With maximal supportive therapy, all patients gradually Mushroom poisoning by Amanita phalloides is a rare but poten- improved from day 3 and recovered without the need for tially fatal disease. The initial symptoms of nausea, vomiting, ab- dominal pain and diarrhea, which are typical for the intoxication, liver transplantation. They were discharged from the can be interpreted as a common gastro-enteritis. The intoxication hospital between 6 to 10 days after admission. can progress to acute liver and renal failure and eventually death. Recognizing the clinical syndrome is extremely important. In this case report, 4 patients with amatoxin intoxication who showed the Discussion typical clinical syndrome are described. The current therapy of amatoxin intoxication is based on small case series, and no ran- Among mushroom intoxications, amatoxin intoxica- domised controlled trials are available. The therapy of amatoxin intoxication consists of supportive care and medical therapy with tion accounts for 90% of all fatalities. Amatoxin poison- silibinin and N-acetylcysteine. Patients who develop acute liver fail- ing is caused by mushroom species belonging to the gen- ure should be considered for liver transplantation. (Acta gastro- era Amanita, Galerina and Lepiota. Amanita phalloides, enterol. belg., 2014, 77, 353-356). commonly known as the “death cap”, causes the majority Key words : amanita phalloides, mushroom poisoning, acute liver of fatal cases.
    [Show full text]
  • Thesis Is Presented for the Degree of Doctor of Philosophy of Curtin University of Technology
    School of Biomedical Sciences Analysis of Candidate Genes within the 3p14-p22 Region of the Human Genome for Association with Bone Mineral Density Phenotypes Benjamin H Mullin This thesis is presented for the Degree of Doctor of Philosophy of Curtin University of Technology February 2011 To the best of my knowledge and belief this thesis contains no material previously published by any other person except where due acknowledgment has been made. This thesis contains no material which has been accepted for the award of any other degree or diploma in any university. Preface The experimental work contained within this thesis was performed in the Department of Endocrinology & Diabetes at Sir Charles Gairdner Hospital under the supervision of Doctor Cyril Mamotte, Associate Professor Scott Wilson, and Professor Richard Prince. All experimental work in this thesis was performed by myself unless otherwise stated. Benjamin H. Mullin, B.Sc. Publications arising from this thesis Mullin, B. H., Prince, R. L., Dick, I. M., Hart, D. J., Spector, T. D., Dudbridge, F. & Wilson, S. G. 2008. Identification of a role for the ARHGEF3 gene in postmenopausal osteoporosis. American Journal of Human Genetics , 82 , 1262-9. Mullin, B. H., Prince, R. L., Mamotte, C., Spector, T. D., Hart, D. J., Dudbridge, F. & Wilson, S. G. 2009. Further genetic evidence suggesting a role for the RhoGTPase-RhoGEF pathway in osteoporosis. Bone , 45 , 387-91. Research grants received during completion of this thesis Wilson, S. G., Prince, R. L., Mamotte C., Mullin B. H. 2008. Influence of the ARHGEF3 gene on bone phenotypes. Arthritis Australia Project Grant ($14,500).
    [Show full text]
  • Peptide Chemistry up to Its Present State
    Appendix In this Appendix biographical sketches are compiled of many scientists who have made notable contributions to the development of peptide chemistry up to its present state. We have tried to consider names mainly connected with important events during the earlier periods of peptide history, but could not include all authors mentioned in the text of this book. This is particularly true for the more recent decades when the number of peptide chemists and biologists increased to such an extent that their enumeration would have gone beyond the scope of this Appendix. 250 Appendix Plate 8. Emil Abderhalden (1877-1950), Photo Plate 9. S. Akabori Leopoldina, Halle J Plate 10. Ernst Bayer Plate 11. Karel Blaha (1926-1988) Appendix 251 Plate 12. Max Brenner Plate 13. Hans Brockmann (1903-1988) Plate 14. Victor Bruckner (1900- 1980) Plate 15. Pehr V. Edman (1916- 1977) 252 Appendix Plate 16. Lyman C. Craig (1906-1974) Plate 17. Vittorio Erspamer Plate 18. Joseph S. Fruton, Biochemist and Historian Appendix 253 Plate 19. Rolf Geiger (1923-1988) Plate 20. Wolfgang Konig Plate 21. Dorothy Hodgkins Plate. 22. Franz Hofmeister (1850-1922), (Fischer, biograph. Lexikon) 254 Appendix Plate 23. The picture shows the late Professor 1.E. Jorpes (r.j and Professor V. Mutt during their favorite pastime in the archipelago on the Baltic near Stockholm Plate 24. Ephraim Katchalski (Katzir) Plate 25. Abraham Patchornik Appendix 255 Plate 26. P.G. Katsoyannis Plate 27. George W. Kenner (1922-1978) Plate 28. Edger Lederer (1908- 1988) Plate 29. Hennann Leuchs (1879-1945) 256 Appendix Plate 30. Choh Hao Li (1913-1987) Plate 31.
    [Show full text]
  • Forest Fungi in Ireland
    FOREST FUNGI IN IRELAND PAUL DOWDING and LOUIS SMITH COFORD, National Council for Forest Research and Development Arena House Arena Road Sandyford Dublin 18 Ireland Tel: + 353 1 2130725 Fax: + 353 1 2130611 © COFORD 2008 First published in 2008 by COFORD, National Council for Forest Research and Development, Dublin, Ireland. All rights reserved. No part of this publication may be reproduced, or stored in a retrieval system or transmitted in any form or by any means, electronic, electrostatic, magnetic tape, mechanical, photocopying recording or otherwise, without prior permission in writing from COFORD. All photographs and illustrations are the copyright of the authors unless otherwise indicated. ISBN 1 902696 62 X Title: Forest fungi in Ireland. Authors: Paul Dowding and Louis Smith Citation: Dowding, P. and Smith, L. 2008. Forest fungi in Ireland. COFORD, Dublin. The views and opinions expressed in this publication belong to the authors alone and do not necessarily reflect those of COFORD. i CONTENTS Foreword..................................................................................................................v Réamhfhocal...........................................................................................................vi Preface ....................................................................................................................vii Réamhrá................................................................................................................viii Acknowledgements...............................................................................................ix
    [Show full text]
  • The Ectomycorrhizal Fungus Amanita Phalloides Was Introduced and Is
    Molecular Ecology (2009) doi: 10.1111/j.1365-294X.2008.04030.x TheBlackwell Publishing Ltd ectomycorrhizal fungus Amanita phalloides was introduced and is expanding its range on the west coast of North America ANNE PRINGLE,* RACHEL I. ADAMS,† HUGH B. CROSS* and THOMAS D. BRUNS‡ *Department of Organismic and Evolutionary Biology, Biological Laboratories, 16 Divinity Avenue, Harvard University, Cambridge, MA 02138, USA, †Department of Biological Sciences, Gilbert Hall, Stanford University, Stanford, CA 94305-5020, USA, ‡Department of Plant and Microbial Biology, 111 Koshland Hall, University of California, Berkeley, CA 94720, USA Abstract The deadly poisonous Amanita phalloides is common along the west coast of North America. Death cap mushrooms are especially abundant in habitats around the San Francisco Bay, California, but the species grows as far south as Los Angeles County and north to Vancouver Island, Canada. At different times, various authors have considered the species as either native or introduced, and the question of whether A. phalloides is an invasive species remains unanswered. We developed four novel loci and used these in combination with the EF1α and IGS loci to explore the phylogeography of the species. The data provide strong evidence for a European origin of North American populations. Genetic diversity is generally greater in European vs. North American populations, suggestive of a genetic bottleneck; polymorphic sites of at least two loci are only polymorphic within Europe although the number of individuals sampled from Europe was half the number sampled from North America. Endemic alleles are not a feature of North American populations, although alleles unique to different parts of Europe were common and were discovered in Scandinavian, mainland French, and Corsican individuals.
    [Show full text]
  • Foraged Mushroom Toxicity Presenting to Urgent Care with Acute Kidney Injury
    CME: This article is offered for AMA PRA Category 1 Credit.™ Visit Case Report https://www.urgentcarecme.com/bundles/jucm-cme for further details. Foraged Mushroom Toxicity Presenting to Urgent Care with Acute Kidney Injury Urgent message: Though it occurs relatively rarely, mushroom toxicity can result in irreversible organ damage and, in certain cases, death if not recognized quickly. Diagnosis can be difficult due to the facts that toxicity may present at different intervals from time of ingestion, de- pending on the species of mushroom, and initial symptoms are nonspecific and similar to those of benign gastrointestinal illnesses. Timely consultation with a poison control center may be life-saving. MICHELE L. STOWE, PA-C, MPAS and JOSHUA W. RUSSELL, MD, MSC, FAAEM, FACEP Introduction ushroom foraging is a popular activity in the U.S. MPacific Northwest (PNW). Based on cultural traditions, many Asian and European immigrants commonly forage for mushrooms, as well. A case of mush room misidentification may occur when a poisonous species in the U.S. is mistaken for an edible species in an indi- vidual’s country of origin, which occurs most commonly among species from Europe and Asia. There are also poi- sonous local native species which can be confused with edible species with similar appearances. Amanita smithi- ana is an example of a poisonous native PNW mush- room which is similar in appearance to, and grows in the same densely forested habitat as, an edible species: the pine mushroom (or matsutake as it is known in Asia), which is used in many traditional Asian dishes.1 Amanita smithiana is known to cause delayed renal failure when ingested, due to the nephrotoxic com- pound allenic norleucine.2 Gastrointestinal symptoms generally begin within 6 hours of ingestion, but renal toxicity does not manifest until 1 to 4 days after inges- ©AdobeStock.com tion; as such, it may not be evident on initial laboratory evaluation.
    [Show full text]
  • A Case of Mushroom Poisoning with Russula Subnigricans: Development of Rhabdomyolysis, Acute Kidney Injury, Cardiogenic Shock, and Death
    CASE REPORT Nephrology http://dx.doi.org/10.3346/jkms.2016.31.7.1164 • J Korean Med Sci 2016; 31: 1164-1167 A Case of Mushroom Poisoning with Russula subnigricans: Development of Rhabdomyolysis, Acute Kidney Injury, Cardiogenic Shock, and Death Jong Tae Cho and Jin Hyung Han Mushroom exposures are increasing worldwide. The incidence and fatality of mushroom poisoning are reported to be increasing. Several new syndromes in mushroom poisoning Department of Internal Medicine, College of have been described. Rhabdomyolytic mushroom poisoning is one of new syndromes. Medicine, Dankook University, Cheonan, Korea Russula subnigricans mushroom can cause delayed-onset rhabdomyolysis with acute Received: 17 April 2015 kidney injury in the severely poisoned patient. There are few reports on the toxicity of R. Accepted: 6 June 2015 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, Address for Correspondence: Jong Tae Cho, MD acute kidney injury, severe hypocalcemia, respiratory failure, ventricular tachycardia, Department of Internal Medicine, College of Medicine, cardiogenic shock, and death. Mushroom poisoning should be considered in the evaluation Dankook University, 201 Manghyang-ro, Dongnam-gu, Cheonan 31116, Korea of rhabdomyolysis of unknown cause. Furthermore, R. subnigricans should be considered E-mail: [email protected] in the mushroom poisoning with rhabdomyolysis. Keywords: Mushroom Poisoning; Rhabdomyolysis; Acute Kidney Injury; Respiratory Failure; Cardiogenic Shock INTRODUCTION in August, 2010 at the Jujak mountain located on the province of Jeollanam­do, the southern area of Korea. He was a bus driv­ More leisure time for hobbies, hiking, and trekking has led to er.
    [Show full text]
  • Toxic Fungi of Western North America
    Toxic Fungi of Western North America by Thomas J. Duffy, MD Published by MykoWeb (www.mykoweb.com) March, 2008 (Web) August, 2008 (PDF) 2 Toxic Fungi of Western North America Copyright © 2008 by Thomas J. Duffy & Michael G. Wood Toxic Fungi of Western North America 3 Contents Introductory Material ........................................................................................... 7 Dedication ............................................................................................................... 7 Preface .................................................................................................................... 7 Acknowledgements ................................................................................................. 7 An Introduction to Mushrooms & Mushroom Poisoning .............................. 9 Introduction and collection of specimens .............................................................. 9 General overview of mushroom poisonings ......................................................... 10 Ecology and general anatomy of fungi ................................................................ 11 Description and habitat of Amanita phalloides and Amanita ocreata .............. 14 History of Amanita ocreata and Amanita phalloides in the West ..................... 18 The classical history of Amanita phalloides and related species ....................... 20 Mushroom poisoning case registry ...................................................................... 21 “Look-Alike” mushrooms .....................................................................................
    [Show full text]
  • |H|||||||||| USOO5278143A United States Patent 19 11 Patent Number: 5,278,143 Shepro Et Al
    |H|||||||||| USOO5278143A United States Patent 19 11 Patent Number: 5,278,143 Shepro et al. (45) Date of Patent: Jan. 11, 1994 (54) PROPHYLACTIC AND THERAPEUTIC OTHER PUBLICATIONS METHODS FOR TREATING NTER LEUKIN-MEDIATED EDEMAS Rudinger, Peptide Hormones, Parsons (Ed.) U. Park t Press, Baltimore, pp. 1-7 (1976). (75) Inventors: David Shepro, Boston, Mass.; J. Doukas et al., Blood, vol. 69, No. 6 pp. 1563-1569 (Jun. Steven Alexander, Nashville, Tenn. 1987). Frimmer, Chem. Abstracts, vol. 71, No. 99937m (1969). 73) Assignee: Trustees of Boston University, Wieland et al., Crit. Rev. Biochem. vol. 5, pp. 185-260 Boston, Mass. (1978). (21) Appl. No.: 807,668 Welbournet al., J. Appl. Physiol 70: 1364-1368 (1991). 22 illed: Dec. 16, 1991 Primary Examiner-Y. Christina Chan 22 Filed ec. 10, Attorney, Agent, or Firm-David Prashker Related U.S. Application Data 57 ABSTRACT m 63 continuation of ser. No. 416,905, Oct. 4, 1989, aban- Unique methods for treating interleukin-mediated ede doned, which is a continuation-in-part of Ser. No. mas in living subjects are provided comprising adminis 47,121, Oct. 4, 1989, abandoned, and a continuation- tering an effective amount of a composition selected in-part of Ser. No. 185,650, Apr. 25, 1988, abandoned. from the group consisting of phallotoxins, phallotoxin analogues, antamanide, or an antananide analogue to 51 Int. Cl. ........................ A61K 37/02; CK /6 the subject. The methods offer prophylactic and thera 52 576.5/35567; 17,353 peutic modes of treatment for both localized and sys s s 8 temic interleukin-mediated edemas. The compositions 58) Field of Search ...................
    [Show full text]
  • Classification of Amanita Species Based on Bilinear Networks With
    agriculture Article Classification of Amanita Species Based on Bilinear Networks with Attention Mechanism Peng Wang, Jiang Liu, Lijia Xu, Peng Huang, Xiong Luo, Yan Hu and Zhiliang Kang * College of Mechanical and Electrical Engineering, Sichuan Agricultural University, Ya’an 625000, China; [email protected] (P.W.); [email protected] (J.L.); [email protected] (L.X.); [email protected] (P.H.); [email protected] (X.L.); [email protected] (Y.H.) * Correspondence: [email protected]; Tel.: +86-186-0835-1703 Abstract: The accurate classification of Amanita is helpful to its research on biological control and medical value, and it can also prevent mushroom poisoning incidents. In this paper, we constructed the Bilinear convolutional neural networks (B-CNN) with attention mechanism model based on transfer learning to realize the classification of Amanita. When the model is trained, the weight on ImageNet is used for pre-training, and the Adam optimizer is used to update network parameters. In the test process, images of Amanita at different growth stages were used to further test the generalization ability of the model. After comparing our model with other models, the results show that our model greatly reduces the number of parameters while achieving high accuracy (95.2%) and has good generalization ability. It is an efficient classification model, which provides a new option for mushroom classification in areas with limited computing resources. Keywords: deep learning; bilinear convolutional neural networks; attention mechanism; trans- fer learning Citation: Wang, P.; Liu, J.; Xu, L.; Huang, P.; Luo, X.; Hu, Y.; Kang, Z.
    [Show full text]
  • The World's Most Poisonous Mushroom, Amanita Phalloides, Is
    Maxwell Moor-Smith, BSc, Raymond Li, BSc(Pharm), MSc, Omar Ahmad, MD, FRCPC The world’s most poisonous mushroom, Amanita phalloides, is growing in BC The expanded range of death cap mushrooms—previously found on the roots of imported European trees but now found in association with native Garry oaks—puts amateur foragers at risk, and recognition of amatoxin poisoning is essential to preventing future fatalities. ABSTRACT: Amatoxins in Amanita acute liver failure occurs. Manage- shown some benefit in a retro- phalloides, commonly known as the ment of the symptomatic patient spective survival analysis. With the death cap mushroom, are responsi- consists of providing supportive expanded range of A. phalloides in ble for 90% of the world’s mushroom- care, promoting renal elimination of BC, physicians should be alert to related fatalities. The most deadly amatoxins, interrupting enterohe- the possibility of amatoxin poison- amatoxin for humans is α-amanitin, patic recirculation of amatoxins, and ing and include it in the differential a bicyclic octapeptide that irrevers- administering proposed antidotes. diagnosis of a patient presenting ibly binds RNA polymerase II, thus Although no established antidote with gastroenteritis or hepatotoxic- preventing protein synthesis and for A. phalloides has been identified, ity and a history of ingesting foraged causing cell death. Three recent N-acetylcysteine and silibinin have mushrooms. poisoning cases in British Columbia show how the death cap can be eas- ily mistaken for edible mushrooms the world’s most poisonous mushroom such as the puffball and the paddy is growing in British Columbia straw mushroom. Since being intro- duced from Europe to the west and mid-Atlantic coasts of North Ameri- Amanita phalloides, commonly known as the death cap mushroom, ca, A.
    [Show full text]
  • Mushrooms on Stamps
    Mushrooms On Stamps Paul J. Brach Scientific Name Edibility Page(s) Amanita gemmata Poisonous 3 Amanita inaurata Not Recommended 4-5 Amanita muscaria v. formosa Poisonous (hallucenogenic) 6 Amanita pantherina Deadly Poisonous 7 Amanita phalloides Deadly Poisonous 8 Amanita rubescens Not Recommended 9-11 Amanita virosa Deadly Poisonous 12-14 Aleuria aurantiaca Edible 15 Sarcocypha coccinea Edible 16 Phlogiotis helvelloides Edible 17 Leccinum aurantiacum Good Edible 18 Boletus parasiticus Not Recommended 19 For this presentation I chose the species for Cantharellus cibarius Choice Edible 20 their occurrence in our 5 county region Cantharellus cinnabarinus Choice Edible 21 Coprinus atramentarius Poisonous 22 surrounding Rochester, NY. My intent is to Coprinus comatus Choice Edible 23 show our stamp collecting audience that an Coprinus disseminatus Edible 24 Clavulinopsis fusiformis Edible 25 artist's rendition of a fungi species depicted Leotia viscosa Harmless 26 on a stamp could be used akin to a Langermannia gigantea Choice Edible 27 Lycoperdon perlatum Good Edible 28-29 guidebook for the study of mushrooms. Entoloma murraii Not Recommended 30 Most pages depict a photograph and related Morchella esculenta Choice Edible 31-32 Russula rosacea Not Recommended (bitter) 33 stamp of the species, along with an edibility Laetiporus sulphureus Choice Edible 34 icon. Enjoy… but just the edible ones! Polyporus squamosus Edible 35 Choice/Good Edible Harmless Not Recommended Poisonous Deadly Poisonous 2 3 4 5 6 7 8 Amanita rubescens (blusher) 9 10
    [Show full text]