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Chemical and biological characterizations of the edible mushroom, volvariella volvacea lectins. / CUHK electronic theses & dissertations collectionJanuary 2004 (has links)
Five novel lectin isoforms, Volvariella volvacea (VV) lectins, designated VVA, VVB, VVC, VVD & VVE, were isolated and purified from the fruiting bodies of an edible mushroom, Volvariella volvacea , by ion-exchange chromatographies in a FPLC system. Their molecular masses are very close, as measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS); they are 12740, 12737, 12709, 12708 & 12707 Da for VVA, VVB, VVC, VVD & VVE, respectively, but the pI values between VVA and the others show distinct differences; the pI value of VVA is around 6.7 and the others are much closer to each other (higher than pI 9.3). Their sugar-binding specificities are thyroglobulin, N-acetylneuramic acid and galacturonic acid. The mitogenic activities of VVA and VVE with distinct pI values were measured using [methyl -3H]thymidine (3H-TdR) incorporation assays, nucleic acid sequence by rapid amplification of cDNA ends analysis, amino acid sequencing and molecular masses by MALDI-TOF/MS and gel electrophoresis, respectively. / VVA and VVE share 98.2% amino acid sequence similarities. Both VVA and VVE are potent mitogens toward mouse CD3+ & CD4 + T-cells, which were mediated through a calcium-dependent activation signaling pathway (Sze et al., 2004). VVA is slightly more effective than VVE in the induction of T cell activation and proliferation, as demonstrated by 3H-TdR incorporation assays, cell flow cytometry for calcium ion mobilization, immunoblotting blot analysis for tyrosine phosphorylation of Lck proteins and Lck shift (p60lck protein), and two dimensional gel electrophoresis for up-regulated proteins. The gene encoding VV lectin was cloned and characterized. The recombinant protein possessed hemagglutinating activity and mitogenic activity, as demonstrated by hemagglutination assays and 3H-TdR incorporation assays respectively. The endoproteinase Arg-C-digested VVA retained the mitogenic activity but lost the hemagglutinating activity, indicating that the mitogenic activity of VVA is not only dependent on the dimerization and tertiary structure of the protein (Paaventhan et al., 2003; Lin et al., 1997), but also on the primary structure of unique amino acid sequences. These endoproteinase fragments have also been used for study of structure-function relationship. / Sze Cho Wing. / "July 2004." / Adviser: Ken W. K. Liu. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0171. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 156-189). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.
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Preparation and structural analysis of non-starch polysaccharides isolated from edible mushrooms.January 1998 (has links)
by Lee Man Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 125-137). / Abstract also in Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT (Chinese version) --- p.v / TABLE OF CONTENTS --- p.vi / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xv / LIST OF ABBREVIATIONS --- p.xvii / Chapter CHAPTER ONE: --- INTRODUCTION --- p.1 / Chapter 1.1 --- Background of mushrooms --- p.1 / Chapter 1.1.1 --- Cultivated mushrooms --- p.1 / Chapter 1.1.1.1 --- Volvariella volvacea --- p.3 / Chapter 1.1.1.2 --- Pleurotus sajor-caju --- p.4 / Chapter 1.1.1.3 --- Pleurotus tuber-regium --- p.5 / Chapter 1.1.2 --- Chemical composition and nutritional value --- p.6 / Chapter 1.1.3 --- Composition and structure of fungal cell wall --- p.9 / Chapter 1.1.4 --- Medicinal attributes of β-glucan in mushrooms --- p.10 / Chapter 1.1.5 --- Structure and antitumor activity β-glucan --- p.11 / Chapter 1.2 --- Dietary fiber --- p.14 / Chapter 1.2.1 --- Composition of dietary fiber --- p.14 / Chapter 1.2.2 --- Preparation of dietary fiber --- p.18 / Chapter 1.3 --- Structural analysis of polysaccharides --- p.19 / Chapter 1.3.1 --- Isolation of polysaccharides --- p.19 / Chapter 1.3.2 --- Methylation analysis --- p.20 / Chapter CHAPTER TWO : --- MATERIALS AND METHODS --- p.22 / Chapter 2.1 --- Sources and preparation of mushroom samples --- p.22 / Chapter 2.1.1 --- V. volvacea --- p.22 / Chapter 2.1.2 --- P. sajor-caju --- p.22 / Chapter 2.1.2.1 --- Fungal strain --- p.22 / Chapter 2.1.2.2 --- Production of spawn --- p.22 / Chapter 2.1.2.3 --- Production of fruiting bodies --- p.23 / Chapter 2.1.3 --- p. tuber-regium --- p.23 / Chapter 2.2 --- Analysis of mushroom composition --- p.24 / Chapter 2.2.1 --- Moisture content --- p.24 / Chapter 2.2.2 --- Starch content --- p.24 / Chapter 2.2.2.1 --- Total glucose --- p.24 / Chapter 2.2.2.2 --- Free glucose --- p.25 / Chapter 2.2.2.3 --- Measurement of glucose content --- p.25 / Chapter 2.2.2.4 --- Total starch content --- p.25 / Chapter 2.2.3 --- Crude protein content --- p.26 / Chapter 2.2.4 --- Amino acid analysis --- p.27 / Chapter 2.3 --- Preparation of mushroom fiber material --- p.28 / Chapter 2.3.1 --- Enzymatic method --- p.28 / Chapter 2.3.1.1 --- Total dietary fiber (TDF) --- p.28 / Chapter 2.3.1.2 --- Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) --- p.29 / Chapter 2.3.2 --- Chemical method --- p.29 / Chapter 2.3.2.1 --- Cell wall material --- p.29 / Chapter 2.4 --- Chemical composition of mushroom fiber material --- p.31 / Chapter 2.4.1 --- Monosaccharide composition of non-starch polysaccharides (NSP) --- p.31 / Chapter 2.4.1.1 --- Acid depolymerisation --- p.31 / Chapter 2.4.1.2 --- Neutral sugar derivatization --- p.31 / Chapter 2.4.1.3 --- Determination of neutral sugars by gas chromatography (GC) --- p.32 / Chapter 2.4.1.4 --- Uronic acid content --- p.32 / Chapter 2.4.2 --- Resistant starch content --- p.33 / Chapter 2.4.3 --- Residual protein content --- p.34 / Chapter 2.5 --- Fractionation of mushroom fiber material --- p.34 / Chapter 2.5.1 --- Solvent extraction --- p.34 / Chapter 2.5.2 --- Anion-exchange chromatography --- p.35 / Chapter 2.5.3 --- Gel permeation chromatography --- p.36 / Chapter 2.6 --- Structural analysis of mushroom fiber material --- p.37 / Chapter 2.6.1 --- Linkage analysis by methylation --- p.37 / Chapter 2.6.1.1 --- Preparation of methylsufinyl carbanion (Dimsyl) --- p.37 / Chapter 2.6.1.2 --- Preparation and dissolution of sample --- p.37 / Chapter 2.6.1.3 --- Methylation --- p.38 / Chapter 2.6.1.4 --- Hydrolysis --- p.38 / Chapter 2.6.1.5 --- Reduction and acetylation --- p.39 / Chapter 2.6.1.6 --- Determination of partially methylated alditol acetate (PMAA) by gas chromatograph-mass spectrometry (GC-MS) --- p.39 / Chapter 2.6.2 --- Fourier-transform infrared (FTIR) spectroscopy --- p.40 / Chapter CHAPTER THREE : --- RESULTS AND DISCUSSION --- p.41 / Chapter 3.1 --- Chemical composition of mushrooms --- p.41 / Chapter 3.1.1 --- Moisture content --- p.41 / Chapter 3.1.2 --- Carbohydrate content --- p.41 / Chapter 3.1.3 --- Protein content --- p.44 / Chapter 3.1.4 --- Amino acid profile --- p.44 / Chapter 3.1.5 --- Dietary fiber content --- p.48 / Chapter 3.1.6 --- Cell wall material --- p.53 / Chapter 3.1.7 --- Comparison of the yield and composition of TDF and CWM --- p.55 / Chapter 3.1.8 --- "Monosaccharide composition of the dietary fiber (TDF, IDF and SDF) and cell wall material (CWM)" --- p.57 / Chapter 3.2 --- Fractionation of TDF and CWM --- p.69 / Chapter 3.2.1 --- Solvent extraction --- p.69 / Chapter 3.2.2 --- Monosaccharide composition of solvent fractionated TDF and CWM --- p.71 / Chapter 3.2.3 --- Anion-exchange chromatography --- p.78 / Chapter 3.2.4 --- Gel permeation chromatography --- p.82 / Chapter 3.2.5 --- Monosaccharide composition of fractionated fiber material by anion-exchange chromatography --- p.84 / Chapter 3.3 --- Structural analysis --- p.86 / Chapter 3.3.1 --- Partially methylated alditol acetate (PMAA) --- p.86 / Chapter 3.3.1.1 --- Alkali-extracted water-soluble fractions of V. volvacea fiber material --- p.95 / Chapter 3.3.1.2 --- Alkali-extracted water-soluble fractions of P. sajor-caju fiber material --- p.99 / Chapter 3.3.1.3 --- Alkali-extracted water-soluble fractions of P. tuber-regium fiber material --- p.102 / Chapter 3.3.1.4 --- Alkali-extracted water-insoluble fractions of the mushroom fiber material --- p.106 / Chapter 3.3.1.5 --- Alkali- and acid- resistant fractions of the mushroom fiber material --- p.109 / Chapter 3.3.2 --- Infrared spectroscopy --- p.112 / Chapter 3.4 --- "β (l→3), (→4) glucan" --- p.119 / Chapter CHAPTER FOUR : --- CONCLUSION --- p.121 / REFERENCES --- p.125 / RELATED PUBLICATIONS --- p.137
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Obsah rtuti v jedlých hubách z lokality historické těžby stříbra / Contents of mercury in edible mushrooms growing in a historical silver-mining areaBÍLÁ, Eva January 2009 (has links)
Contents of mercury were determined using AAS method (instrument AMA-254) in 109 fruiting body samples of 14 edible mushroom species. The mushrooms were collected from a forest of a historical area of silver mining. The highest mercury accumulation was observed in samples of Boletus aestivalis, Maccrolepiota procera and Boletus edulis. Mercury content was highly comparable with concentrations observed in unpolluted sites.
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Crescimento micelial, produção e características bromatológicas do shiitake em função de linhagens e de propriedades físicas e químicas de espécies e clones de eucaliptoAndrade, Meire Cristina Nogueira de [UNESP] 24 May 2007 (has links) (PDF)
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andrade_mcn_dr_botfca.pdf: 1132419 bytes, checksum: 28616e10ef21df6298a36b3b132aeeb9 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Mycelium growth, production and bromatologicals characteristics of shiitake in function of lineages and chemical and physical properties of eucalyptus clones and species were evaluated. In Experiment 1, mycelium growth of two Lentinula edodes (Berk.) Pegler (LE-95/01 and LE-96/18) species in culture mediums prepared with sawdust extract from seven species (E. saligna, E. grandis, E. urophylla, E. camaldulensis, E. citriodora, E. paniculata e E. pellita) and three eucalyptus clones (hybrid E. grandis x E. urophylla) was analyzed. The experimental design was totally randomized, in 2x10 factorial design, totalizing 20 treatments with 10 repetitions, being that each repetition corresponded to one Petri dish. In Experiment 2, mycelium growth of eight L. edodes lineages (LE-96/17, LE-95/02, LE-95/07, LE-98/55, LE-96/18, LE-95/01, LE-96/13 and LE-98/47) in culture mediums prepared with sawdust extract from Eucalyptus spp was evaluated. The experimental design was totally randomized, with 8 treatments and 8 repetitions, being that each repetition corresponded to one Petri dish. In Experiment 3, production and bromatological characterization of two Lentinula edodes lineages cultivated in seven species and three clones of eucalyptus was evaluated. The experimental design was totally randomized, in 2x10 factorial design, totalizing 20 treatments with 40 repetitions, being that each repetition corresponded to one log. In Experiment 4, physical and chemical properties of seven species and three clones of eucalyptus before and after the cultivation of two L. edodes lineages was evaluated. The experimental design was totally randomized, in 2x10 factorial design, totalizing 20 treatments with 9 repetitions, being that each repetition corresponded to one log. The culture medium that provided highest averages of mycelium growth of L. edodes lineages LE-95/01 and LE-96/18 was the one with... (Complete abstract click electronic access below)
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Produtividade de Agaricus blazei Murril em função do ambiente de cultivo, massa do substrato e camada de coberturaBraga, Gilberto Costa [UNESP] 20 April 1999 (has links) (PDF)
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braga_gc_dr_botfca.pdf: 432107 bytes, checksum: 7b22fe235f002c836b40e7a4ee958ec6 (MD5) / Dado à escassez de informações ou relatos técnico-científicos sobre o cultivo protegido de Agaricus blazei Murril, este trabalho teve como objetivos, avaliar os efeitos do ambiente de cultivo, da massa do substrato, da espessura da camada de cobertura e de choques térmicos no substrato de cultivo, na frutificação e produtividade de cogumelos A. blazei. Foram realizados dois experimentos. O primeiro constou da avaliação de três espessuras da camada de cobertura (3, 5 e 8 cm), duas massas de substrato (5 e 10kg) e dois ambientes de cultivo protegido (ambiente coberto com bambu e estufa coberta com lona plástica). O substrato de cultivo foi acondicionado em caixas plásticas, representando as parcelas experimentais. Cada tratamento foi constituído por 5 parcelas experimentais. O segundo experimento foi constituído por 5 tratamentos, sendo 3 com choques térmicos de 15oC e 2 controles (15oC e 25oC). Os choques térmicos foram aplicados no substrato de cultivo variando o número de choques e a duração de cada um, sendo tratamento 1: três choques térmicos com duração de 24 horas cada e em intervalos de 21 dias; tratamento 2: três choques térmicos com duração de 72 horas cada e em intervalos de 21 dias e tratamento 3: um choque térmico com duração de 72 horas. Nesse experimento, o cultivo foi realizado em uma sala escura sob condições controladas de temperatura (25oC) e umidade relativa do ar (80%). Os resultados foram obtidos a partir de 4 períodos de 21 dias de mensurações, que constituíam os intervalos entre os choques. Em função desses dois experimentos, foi realizada uma terceira avaliação onde se determinou a produtividade máxima, a taxa máxima de produtividade e o tempo de início de produção, em função dos três ambientes de cultivo referidos nesses dois experimentos. A partir dos resultados, concluiu-se que: no inverno, o ambiente coberto com plástico... / Given the shortage of information and technical scientific reports on the protected cultivation of A. blazei, the objectives of this work were to evaluate the effects of cultivation environment, mass of the substrate, thickness of the casing layer, and thermal shocks in the cultivation substrate, on the growth and productivity of the mushroom A. blazei Murril. To obtain the results, two experiments were performed. The first consisted of evaluating three thicknesses of casing layer (3, 5 and 8 cm), two substrates of different masses (5 and 10kg) and two types of protected cultivation (covered with bamboo and in a greenhouse covered with blue plastic canvas). in plastic boxes. Each different treatment was performed on 5 units (plastic boxes) of cultivation substrate. The second experiment consisted of five units, three with thermal shocks and one a control. The thermal shocks were applied to the cultivation substrate in varying number and duration, treatment 1 being: three thermal shocks of 24 hours duration; treatment 2: three thermal shocks of 72 hours duration, and treatment 3: one thermal shock of 72 hours duration. In this experiment, cultivation was carried out under controlled conditions in a room at temperature (25oC) and relative humidity (80%). The measurements were taken and shocks applied at four 21 day intervals. After these two experiments, a third evaluation was accomplished determining the maximum productivity, the maximum rate of production and the beginning time of production, in relation to the three cultivation environments used in the two experiments. From these results, it was concluded that: in the winter, the greenhouse covered with plastic, promoted more favourable environmental conditions for the production of A. blazei, than the structure covered with bamboo; in the greenhouse, casing layers of 5 and 8 cm thickness were more productive than the 3 cm layer... (Complete abstract click electronic access below)
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Estudo comparativo de diferentes cogumelos comestíveis comercializados no Brasil: ação contra radicais livres, espécies reativas e atividade antibacterianaKamila Maria Silveira Negri [UNESP] 20 January 2012 (has links) (PDF)
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000694406.pdf: 795605 bytes, checksum: 54ce9cb35056780b7a687b4fcd698bfb (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O excesso na geração de espécies reativas como os radicais livres pode resultar num desequilíbrio que, embora benéfico em situações como o combate a micro-organismos patógenos, está implicado com diversas patologias crônicas e com o envelhecimento. Assim é comum a indicação de compostos antioxidantes para a prevenção destas patologias e inclusive, algumas pesquisas com substâncias antioxidantes já resultaram no desenvolvimento de novos medicamentos. Desta forma, o estudo de fontes naturais que apresentam substâncias ativas sobre as diferentes espécies oxidantes geradas em nosso organismo é de grande importância, visto as implicações destas em várias patologias. Entre estas fontes, Agaricus blazei Murrill é um cogumelo comestível e medicinal, popularmente utilizado na prevenção do envelhecimento e no tratamento de diversas doenças, como câncer, artrite reumatóide, artrose, aterosclerose, diabetes e hipertensão. Tem sido relatado também, o uso de outros cogumelos como Agaricus bisporus (champignon), Lentinus edodes (shiitake) e Pleurotus ostreatus (shimeji ou hiratake), de forma similar ao Agaricus blazei. Além da função nutricional desse cogumelo é importante verificar possíveis atividades biológicas, o que poderia render novos produtos com alegações de propriedades funcionais ou terapêuticas, como as já existentes para o A. blazei. Assim, este projeto objetivou o estudo do perfil de atividade antioxidante (químico, enzimático e ex vivo) e análise de atividade antibacteriana de três cogumelos cultivados no Brasil em comparação com o Agaricus blazei Murrill. Foram utilizadas metodologias de análise de atividade antioxidante como os ensaios de ação scavenger do DPPH·, ABTS·+, O2 ·-, NO, H2O2 e HOCl, e também por análise do sistema oxidativo catalisado por mieloperoxidase (MPO) e... / The excessive generation of reactive oxygen species, like free radicals, can lead to an imbalance which, although benefical in such situations to combat pathogenic microorganisms, is involved with various chronic diseases and aging. Thus, it is common to indicate the use of antioxidants for prevention of these diseases, and some research about antioxidant products have already resulted in the development of new medicines. Thus, the study of natural sources that contain active compounds over the different oxidant species, generated in our bodies, is of great importance, due their implications in various pathologies. Among these sources, Agaricus blazei Murrill is an edible and medicinal mushroom, commonly used in the prevention of aging and the treatment of various diseases such as cancer, rheumatoid arthritis, osteoarthritis, atherosclerosis, diabetes and hypertension. It has also been reported, the use of other Agaricus bisporus (champignon), Lentinus edodes (shiitake) and Pleurotus ostreatus (shimeji or hiratake), similar to Agaricus blazei. Besides its nutritional properties, it is important to check possible biological activities, which could render functional property claims as those existing for the A. blazei. Thus, mushrooms such as this project aims to study the profile of antioxidant activity (chemical, enzymatic and ex vivo) of mushrooms cultivated in Brazil, comparing to Agaricus blazei Murrill. Antioxidant activities were evaluated through DPPH·, ABTS·+, O2 ·-, NO, H2O2 and HOCl and also by analysis of the oxidative system catalyzed by myeloperoxidase (MPO) and horseradish peroxidase (HRP). In relation to cellular analysis and antibacterial, it was determined toxicity erythrocytes and neutrophils, and activity in relation to Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa... (Complete abstract click electronic access below)
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Produção, economicidade e parâmetros energéticos do cogumelo Agaricus blazei: um enfoque de cadeia produtiva /Herrera, Ozana Maria, 1956- January 2001 (has links)
Orientador: Maristela Simões do Carmo / Resumo: Nos últimos anos tem sido intensa a busca por atividades agrícolas que conciliem a produção de alimentos com preservação ambiental e viabilidade econômica e social. A globalização modificou as relações econômicas de forma ampla e definida, obrigando todos os setores da economia a realizarem rápidas adaptações para garantir sua participação e sobrevivência no mercado. Essas transformações impactam, de forma gradual e constante, a organização da produção agroindustrial. A eficiência, nos sistemas agroindustriais, provém de relações entre preços e produtividade dos fatores de produção (terra, capital e trabalho) e da tecnologia. São importantes também a minimização dos custos de transação ao longo das cadeias, e um eficiente sistema de coordenação ou de governança. Por outro lado, os cogumelos medicinais podem ser importantes aliados na resolução de muitos problemas, desde o aproveitamento de resíduos agroindustriais até a produção de medicamentos utilizados no combate à diversas doenças. O Agaricus blazei é um cogumelo originário da Mata Atlântica brasileira e vem ganhando credibilidade como produto nutracêutico e medicinal em mercados internacionais...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In the last years it has been intense the search for agricultural activities that reconcile the production of foods with environmental preservation and economical and social viability. The globalization modified the economical relationship in a wide and defined way, forcing all of the sections of the economy to perform it fast adaptations to guarantee their participation and survival in the market. Those transformations impact, in a gradual and constant way, the organization of the agroindustrial production. The efficiency in the agroindustrial system, comes from relationships between prices and productivity of the production factors (land, capital and work), from technology, and others factors analyzed traditionally, derived of the minimization of the cost transaction along the chains, and of an efficient coordination or governance system. Otherwise, the medicinal mushrooms can be important allies in the resolution of many problems, which go from the utilization of agroindustrial residue to the production of medicaments used to fight several diseases. Agaricus blazei is a mushroom...(Complete abstract, click electronic access below) / Doutor
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Obsah rtuti v houbách rostoucích poblíž Horního Slavkova. / Mercury contents in mushrooms growing nearby Horní Slavkov.BAŽILOVÁ, Štěpánka January 2008 (has links)
Contents of mercury were determined using AAS method (instrument AMA-254) in 130 fruiting body samples of 13 edible mushroom species. The mushrooms were collected from a forest of a historical area of tin mining. The highest mercury accumulation was observed in samples of Boletus edulis and Calocybe gambosa. Mercury content was highly comparable with concentrations observed in unpolluted sites.
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Obsah rtuti v houbách z území poblíž závodu na výrobu střeliva a průmyslové střelnice / Mercury contents in mushrooms growing nearby a factory producing ammunition and an industrial shooting-rangeBRÁZDOVÁ, Lenka January 2008 (has links)
Contents of mercury were determined using AAS method (instrument AMA-254) in 119 fruiting body samples of 41 mostly edible mushroom species. The mushrooms were collected from a forest near by the ammunition factory and its industrial shooting-range (fy Sellier & Bellot, Vlašim, Czech Republic). The highest mercury accumulation was observed in samples of Agaricus arvensis, Lepista nuda, Calocybe gambosa and Russu-la nigricans. Mercury content was highly comparable with concentrations observed in unpolluted sites.
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Obsah rtuti v hubách z území poblíž nádrže Skalka / Mercury contents in mushrooms growing nearby a reservoir SkalkaSZABOVÁ, Martina January 2010 (has links)
Contents of mercury were determined using AAS method (instrument AMA-254) in 64 fruiting body samples of 15 edible mushroom species. The mushrooms were collected from a forest nearby a reservoir Skalka. The highest mercury accumulation was observed in a sample of Boletus edulis (15.5 mg/ kg dry matter). Mercury contents were highly comparable with concentrations observed in unpolluted sites. It can be therefore concluded that mercury contents in mushroom from the observed area do not represent any health risk.
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