Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms

Lisa J Bazzle,1 Marc A Cubeta,2 Steven L Marks,1 David C Dorman3 1Department of Clinical Sciences, College of Veterinary Medicine, 2Department of Plant Pathology, College of Agriculture and Life Sciences, Center for Integrated Fungal Research, 3Department of Molecular and Biomedical Sciences, Colle...

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Autores principales:	Bazzle LJ, Cubeta MA, Marks SL, Dorman DC
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2014
Materias:	Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/315d5c9615314a4481c5e8714d81cb2f
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spelling	oai:doaj.org-article:315d5c9615314a4481c5e8714d81cb2f2021-12-02T05:35:49ZFeasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms2230-2034https://doaj.org/article/315d5c9615314a4481c5e8714d81cb2f2014-12-01T00:00:00Zhttp://www.dovepress.com/feasibility-of-flotation-concentration-of-fungal-spores-as-a-method-to-peer-reviewed-article-VMRRhttps://doaj.org/toc/2230-2034 Lisa J Bazzle,1 Marc A Cubeta,2 Steven L Marks,1 David C Dorman3 1Department of Clinical Sciences, College of Veterinary Medicine, 2Department of Plant Pathology, College of Agriculture and Life Sciences, Center for Integrated Fungal Research, 3Department of Molecular and Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Purpose: Mushroom poisoning is a recurring and challenging problem in veterinary medicine. Diagnosis of mushroom exposure in animals is hampered by the lack of rapid diagnostic tests. Our study evaluated the feasibility of using flotation concentration and microscopic evaluation of spores for mushroom identification. Evaluation of this method in living animals exposed to toxigenic mushrooms is limited by ethical constraints; therefore, we relied upon the use of an in vitro model that mimics the oral and gastric phases of digestion. Methods: In our study, mycologist-identified toxigenic (poisonous) and nontoxigenic fresh mushrooms were collected in North Carolina, USA. In phase 1, quantitative spore recovery rates were determined following magnesium sulfate, modified Sheather's sugar solution, and zinc sulfate flotation (n=16 fungal species). In phase 2, mushrooms (n=40 fungal species) were macerated and digested for up to 2 hours in a salivary and gastric juice simulant. The partially digested material was acid neutralized, filtered, and spores concentrated using zinc sulfate flotation followed by microscopic evaluation of spore morphology. Results: Mean spore recovery rates for the three flotation fluids ranged from 32.5% to 41.0% (P=0.82). Mean (± standard error of the mean) Amanita spp. spore recovery rates were 38.1%±3.4%, 36.9%±8.6%, and 74.5%±1.6% (P=0.0012) for the magnesium sulfate, Sheather's sugar, and zinc sulfate solutions, respectively. Zinc sulfate flotation following in vitro acid digestion (phase 2) yielded spore numbers adequate for microscopic visualization in 97.5% of trials. The most common spore shapes observed were globose, spiked, elliptical, smooth and reticulate. Conclusion: Flotation can concentrate mushroom spores; however, false negative results can occur. Spore morphology could not be used to differentiate species of mushroom-forming fungi since the spore shape and surface characteristics seen in the present study were often observed with multiple species of mushroom-forming fungi. Keywords: gastrointestinal contents, mushroom spore identification, mushroom toxicity, Amanita spp.Bazzle LJCubeta MAMarks SLDorman DCDove Medical PressarticleVeterinary medicineSF600-1100ENVeterinary Medicine: Research and Reports, Vol 2015, Iss default, Pp 1-9 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Veterinary medicine SF600-1100
spellingShingle	Veterinary medicine SF600-1100 Bazzle LJ Cubeta MA Marks SL Dorman DC Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
description	Lisa J Bazzle,1 Marc A Cubeta,2 Steven L Marks,1 David C Dorman3 1Department of Clinical Sciences, College of Veterinary Medicine, 2Department of Plant Pathology, College of Agriculture and Life Sciences, Center for Integrated Fungal Research, 3Department of Molecular and Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Purpose: Mushroom poisoning is a recurring and challenging problem in veterinary medicine. Diagnosis of mushroom exposure in animals is hampered by the lack of rapid diagnostic tests. Our study evaluated the feasibility of using flotation concentration and microscopic evaluation of spores for mushroom identification. Evaluation of this method in living animals exposed to toxigenic mushrooms is limited by ethical constraints; therefore, we relied upon the use of an in vitro model that mimics the oral and gastric phases of digestion. Methods: In our study, mycologist-identified toxigenic (poisonous) and nontoxigenic fresh mushrooms were collected in North Carolina, USA. In phase 1, quantitative spore recovery rates were determined following magnesium sulfate, modified Sheather's sugar solution, and zinc sulfate flotation (n=16 fungal species). In phase 2, mushrooms (n=40 fungal species) were macerated and digested for up to 2 hours in a salivary and gastric juice simulant. The partially digested material was acid neutralized, filtered, and spores concentrated using zinc sulfate flotation followed by microscopic evaluation of spore morphology. Results: Mean spore recovery rates for the three flotation fluids ranged from 32.5% to 41.0% (P=0.82). Mean (± standard error of the mean) Amanita spp. spore recovery rates were 38.1%±3.4%, 36.9%±8.6%, and 74.5%±1.6% (P=0.0012) for the magnesium sulfate, Sheather's sugar, and zinc sulfate solutions, respectively. Zinc sulfate flotation following in vitro acid digestion (phase 2) yielded spore numbers adequate for microscopic visualization in 97.5% of trials. The most common spore shapes observed were globose, spiked, elliptical, smooth and reticulate. Conclusion: Flotation can concentrate mushroom spores; however, false negative results can occur. Spore morphology could not be used to differentiate species of mushroom-forming fungi since the spore shape and surface characteristics seen in the present study were often observed with multiple species of mushroom-forming fungi. Keywords: gastrointestinal contents, mushroom spore identification, mushroom toxicity, Amanita spp.
format	article
author	Bazzle LJ Cubeta MA Marks SL Dorman DC
author_facet	Bazzle LJ Cubeta MA Marks SL Dorman DC
author_sort	Bazzle LJ
title	Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
title_short	Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
title_full	Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
title_fullStr	Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
title_full_unstemmed	Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
title_sort	feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms
publisher	Dove Medical Press
publishDate	2014
url	https://doaj.org/article/315d5c9615314a4481c5e8714d81cb2f
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Feasibility of flotation concentration of fungal spores as a method to identify toxigenic mushrooms

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